ORT Supplement 3/2016 DGHO Abstractband 2016 by Karger Publishers

Oncol Res Treat 39(suppl 3) XIV + 350 (2016)

39 | S3 | 16

print online ISSN 2296–5270 e-ISSN 2296–5262 ISBN 978-3-318-05959-5

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Band 39, Supplement 3, Oktober 2016

Jahrestagung der D eutschen, Österreichischen und Schweizerischen Gesellschaften für Hämatologie und Medizinische Onkologie Leipzig, 14.–18. Oktober 2016

ABSTRACTS Herausgeber

Andreas Hochhaus, Jena

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CONTENTS AUTHOR INDEX

Band 39, Supplement 3, Oktober 2016

Offizielles Organ von DGHO – Deutsche Gesellschaft für Hämatologie und Medizinische Onkologie OeGHO – Österreichische Gesellschaft für Hämatologie & Medizinische Onkologie DFaG – Deutsche Fatigue Gesellschaft AIO – Arbeitsgemeinschaft Internistische Onkologie in der Deutschen Krebsgesellschaft e.V. SGH-SSH – Schweizerische Gesellschaft für Hämatologie Mitglied der Deutschen Krebsgesellschaft e.V.

Editors Editor-in-Chief

Associate Editors

M. Hallek, Köln

S. Al-Batran, Frankfurt/M. C. Berking, München C. Bokemeyer, Hamburg M. Borner, Bern T. Cerny, St. Gallen H. T. Eich, Münster A. Engert, Köln M. Fassnacht, Würzburg F. Geiser, Bonn B. Groner, Frankfurt/M. V. Heinemann, München M. Hentrich, München R. D. Issels, München

W. Janni, Ulm U. R. Kleeberg, Hamburg A. A. Lammertsma, Amsterdam H. Lang, Mainz M. Moehler, Mainz P. Reichardt, Berlin M. Schuler, Essen R. Stupp, Zürich M. Theobald, Mainz R. Thomas, Köln U. Wedding, Jena J. A. Werner, Marburg O. Zivanovic, New York

A. Heidenreich, Aachen U. Herrlinger, Bonn A. Hochhaus, Jena R.-D. Hofheinz, Mannheim F. Honecker, St. Gallen R. D. Issels, München V. Jacobs, Salzburg K. Jordan, Halle U. Keilholz, Berlin J. P. Klussmann, Gießen H. Kölbl, Wien W. Kuhn, Bonn

H.-J. Lenz, Los Angeles P. Mallmann, Köln H. Moch, Zürich S. Reske, Ulm I. Runnebaum, Jena P. Schöffski, Leuven C. Spitzweg, München I. Strohscheer, St. Peter-Ording S. Ugurel, Essen R. Voltz, Köln M. Weller, Zürich

Editorial Board P. Albers, Düsseldorf C. Bausewein, München L. Bergmann, Frankfurt/M. J. Boos, Münster P. Brossart, Bonn W. Budach, Düsseldorf R. Büttner, Bonn J. Debus, Heidelberg E. Dippel, Ludwigshafen A. Du Bois, Essen T. Fehm, Düsseldorf N. Harbeck, München

Editorial Office S. Karger GmbH Attn. Dr. Steffi Hentzelt P.O. Box D-79095 Freiburg E-mail s.hentzelt@karger.com

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CONTENTS AUTHOR INDEX

Oncol Res Treat 2016;39(suppl 3)

Tagungspräsident

Prof. Dr. Andreas Hochhaus, Jena

Wissenschaftliches Sekretariat

Prof. Dr. Paul Graf La Rosée, Villingen-Schwenningen Dr. Ekkehard Eigendorff, Jena Dr. Tobias Rachow, Jena

Abstractgutachter Bernd Alt-Epping, Claudia Baldus, Walter Baumann, Wolfgang Berdel, Lothar Bergmann, Mascha Binder, Frank-Dietmar Böhmer, Carsten Bokemeyer, Peter Borchmann, Markus Maximilian Borner, Martin Bornhäuser, Jan Braess, Peter Brossart, Tim Hendrik Brümmendorf, Christian Buske, Nathan Cantoni, Richard Cathomas, Thomas Cerny, Yves Chalandon, Joachim Clement, Stefan Diem, Uta Dirksen, Christian Dittrich, Konstanze Döhner, Martin Dreyling, Ulrich Dührsen, Alexander Egle, Barbara Eichhorst, Ekkehard Eigendorff, Wolfgang Eisterer, Monika Engelhardt, Geneviève Favre, Thomas Fischer, Gunnar Folprecht, Norbert Frickhofen, Michael Fridrik, Michael Fuchs, Alois Günther Gastl, Thomas Gauler, Armin Gerger, Bertram Glaß, Nicola Gökbuget, Hartmut Goldschmidt, Katharina Götze, Ullrich Graeven, Paul Graf La Rosée, Hildegard Greinix, Martin Griesshammer, Viktor Grünwald, Tayfun Güngör, Susanna Hegewisch-Becker, Florian Heidel, Dominik Heim, Volker Heinemann, Wolfgang Herr, Pia Heußner, Inken Hilgendorf, Felicitas Hitz, Andreas Hochhaus, Britta Höchsmann, Silvia Hofer, Ralf-Dieter Hofheinz, WolfKarsten Hofmann, Jutta Hübner, Paul Imbach, Ulrich Jäger, Dirk Jäger, Martin Janz, Karin Jordan, Constanze Junghans, Christian Junghanß, Lothar Kanz, Ursula Kapp, Felix Keil, Ulrich Keilholz, Michael Kiehl, Alexander Kiss, Marianne Kloke, Michael Kneba, Maren Knödler, Matthias Kochanek, Michael Köhler, Gerald Kolb, Michael Krainer, Stefan Krause, Frank Kroschinsky, Volker Kunzmann, Florian Langer, Claudia Lengerke, Eva Lengfelder, Georg Lenz, Anne Letsch, Udo Lindig, Lars H. Lindner, Hartmut Link, Sonja Loges, Florian Lordick, Heinz Ludwig, Diana Lüftner, Andreas Mackensen, Christoph Mamot, Markus G. Manz , Norbert Marschner, Georg Maschmeyer, Axel Matzdorff, Hans-Günther Mergenthaler, Georgia Metzgeroth, Lars-Olof Mügge, Andreas Müller, Lothar Müller, Carsten Müller-Tidow, Urban Novak, Karin Oechsle, Helmut Oettle, Friedrich Overkamp, Ingrid Pabinger, Jakob Passweg, Christian Peschel, Andrea Petermann-Meyer, Andreas Petzer, Michael Pfeilstöcker, Robert Pirker, Uwe Platzbecker, Lisa Pleyer, Tobias Pukrop, Julia Quidde, Peter Reichardt, Andreas Reiter, Christoph Renner, Hanno Riess, Christoph Röllig, Alexander Röth, Thomas Ruhstaller, Mathias J. Rummel , Niklaus Schäfer, Werner Scheithauer, Peter Schellongowski, Jan Schildmann, Kristina Schilling, Harald Schmalenberg, Manuela Schmidinger, Norbert Schmitz, Sebastian Scholl, Karin Schrenk, Martin Schuler, Ulrich Schuler, Ulf Seifart, Heinz Sill, Ernst Späth-Schwalbe, Michael Stahl, Frank Stenner-Liewen, Michael Steurer, Georg Stüssi, Hans Tesch, Josef Thaler, Matthias Theobald, Alexandre Theocharides, Michael Thomas, Peter Thuß-Patience, Andreas Tiede, Martin Trepel, Lorenz Trümper, Christian Urban, Maria Vehreschild, Arndt Vogel, Gunhild von Amsberg, Marie von Lilienfeld-Toal, Roger von Moos, Claudia Waskow, Herbert Watzke, Ulrich Wedding, Katja Weisel, Clemens-Martin Wendtner, Martin Wilhelm, Wolfgang Willenbacher, Andreas Willer, Eva Winkler, Mathias Witzens-Harig, Jürgen Wolf, Albert Wölfler, Ewald Wöll Angaben ohne Gewähr

Disclosure Statement The editor declares no conflict of interest.

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CONTENTS AUTHOR INDEX

Oncol Res Treat 2016;39(suppl 3)

Jahrestagung der D eutschen, Österreichischen und Schweizerischen Gesellschaften für Hämatologie und Medizinische Onkologie

Band 39, Supplement 3, Oktober 2016

Leipzig, 14.– 18. Oktober 2016

ABSTRACTS Herausgeber

Andreas Hochhaus, Jena

Der Veranstalter des wissenschaftlichen Kongresses, der Verein zur Förderung der Weiterbildung in der Hämatologie und Onkologie e.V. übernimmt keine Gewähr für die Richtigkeit der Angaben in den Abstracts. Beiträge und Anzeigen geben nicht notwendigerweise die Auffassung der Vorstände wieder.

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CONTENTS AUTHOR INDEX

Contents ∙ Inhalt ∙ (chronologisch) Oncol Res Treat 2016;39(suppl 3)

Abstract-No. Page-No.

Eröffnungsveranstaltung Fortbildung Management der chronischen myeloischen Leukämie Gynäkologische Tumoren Palliativmedizin Freier Vortrag Management hämatologischer Erkrankungen älterer Patienten Fortbildung Kopf-Hals-Tumoren Freier Vortrag Infektionsmanagement Rehabilitation Chronische lymphatische Leukämie – Biologie Fortbildung Indolente Non-Hodgkin-Lymphome CUP-Syndrom Lebertumoren Wissenschaftliches Symposium Neue immuntherapeutische Ansätze nach allogener Stammzelltransplantation ALL: Molekulare und zelluläre Grundlagen für zielgerichtete Therapien Fortbildung Hirnmetastasen und ZNS-Lymphome Thrombophilie Freier Vortrag Zellbiologie Infektionen / supportive Therapie Fortbildung Interprofessionelle Sitzung: Geriatrische Onkologie (für Ärzte und Pflegekräfte) Freier Vortrag Mastozytose, myeloproliferative Neoplasien Fortbildung Immuntherapie in der Hämatologie und Onkologie Mammakarzinom: (Neo)adjuvante Therapie ZNS-Tumoren Kontroverse Indikationen zur Stammzelltransplantation Lebensqualitäts-Assessment Eosinophilie, Mastozytose Freier Vortrag Nichtkleinzelliges Lungenkarzinom Fortbildung Interprofessionelle Sitzung: Therapiebegrenzung (für Ärzte und Pflegekräfte) Freier Vortrag Chronische lymphatische Leukämie – Therapie Fortbildung Intensivmedizin und Hämatologie/Onkologie Freier Vortrag Chronische myeloische Leukämie – Erstlinien- und Absetzstudien Wissenschaftliches Symposium Neue Optionen beim Hodgkin-Lymphom Fortbildung Standardisierte Leukämie-Diagnostik Freier Vortrag Versorgungsforschung Akute myeloische Leukämie – Therapie 1, Mutationsprofil Posterdiskussion Akute myeloische Leukämie Chronische myeloische Leukämie Myelodysplastisches Syndrom, sonstige Hämatologie Multiples Myelom 1 Lymphome Allogene Stammzelltransplantation 1 Immuntherapie Lungentumoren Kolorektale Karzinome

V14 V15 V25 V27–V28 V29–V34 V35 V42–V47 V48–V53 V56–V61 V63–V65 V66–V67 V69

1 1 1 1 2 6 7 9 11 14 15 15

V75

V77–V79 V80–V82 V85 V86–V91 V92–V97

16 17 18 18 20

V100–V101 V103–V108 V110–V112 V114 V117 V119–V122 V123–V126 V127–V130 V131–V135

23 24 27 27 27 28 29 30 31

V138–V140 V141–V146 V150–V151 V153–V158 V160 V168–V169 V170–V175 V178–V183 P186–P200 P201–P215 P216–P226 P227–P241 P242–P256 P257–P271 P272–P285 P286–P300 P301–P312

33 34 36 37 40 40 41 44 46 52 59 64 70 76 82 87 93

Plenarsitzung

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CONTENTS AUTHOR INDEX

Contents ∙ Inhalt ∙ (chronologisch) Oncol Res Treat 2016;39(suppl 3)

Abstract-No. Page-No.

Myelodysplastisches Syndrom Chronische lymphatische Leukämie Sarkome 2016 – was gibt es Neues? Melanom Blut ist ein besonderer Saft – Transfusionsmedizin Wissenschaftliches Symposium Kontroversen in der Therapie des kolorektalen Karzinoms Fortbildung Nierenzellkarzinom Freier Vortrag Immuntherapie solider Tumoren Epigenetik Expertenseminar Patienten mit ZNS-Lymphomen Freier Vortrag Akute myeloische Leukämie – Therapie 2 Fortbildung Multiples Myelom Pankreaskarzinom Wissenschaftliches Symposium Wie teuer dürfen Arzneimittel sein? Fortbildung Noch kurativ oder schon palliativ? Ethik in der Onkologie Schwangerschaft und Krebs Freier Vortrag Mammakarzinom Niedrigmaligne B-Zell-Lymphome – Klinik Fortbildung Migrationsanämien Freier Vortrag Myelodysplastisches Syndrom – experimentell Debatten CLL -Zukunft ohne Chemotherapie ? Fortbildung Innovation durch Kooperation – Die Deutsche CML-Allianz Update Infektionen in der Hämatologie und Onkologie – 20 Jahre AGIHO Aggressive Non-Hodgkin-Lymphome Magenkarzinome Freier Vortrag Graft-versus-Host-Disease Expertenseminar Diagnostische und therapeutische Fallstricke bei MPN-assoziierter Thrombophilie Fortbildung Management myeloproliferativer Neoplasien Management des Hodgkin-Lymphoms Wissenschaftliches Symposium Therapeutische Targets bei AML – Vision und Wirklichkeit Fortbildung Supportive Therapien: aktuelle Leitlinien in der onkologischen Praxis Freier Vortrag Immuntherapie von Leukämien und Lymphomen Nicht maligne Hämatologie (Minimale) Resterkrankung nach Leukämietherapie Wissenschaftliches Symposium Wirkungen und Nebenwirkungen im Versorgungsalltag Debatten Ist die Zeit reif für eine therapiefreie Remission bei der CML? Posterdiskussion Akute myeloische Leukämie, experimentell Myeloproliferative Neoplasien, Gerinnung Aggressive Non-Hodgkin-Lymphome Multiples Myelom 2 Allogene Stammzelltransplantation 2 Lungentumoren, Kopf-Hals-Tumoren Palliativmedizin, Integrative Onkologie Gynäkologische Tumoren, Mammakarzinom Kasuistiken – Hämatologie

V313–V316 V319 V321–V323 V327 V329 V335 V336–V337 V339–V344 V345–V350 V352 V353–V358 V359–V361 V367 V370–V371 V374–V377 V381 V384–V389 V390–V395 V396–V398 V401–V406 V411 V414–V418

98 99 100 100 101 101 101 102 106 109 109 112 113 114 114 115 116 118 121 122 125 125

V421 V430–V432 V434–V435 V437–V422

127 127 127 128

V444 V451–V453 V454 V458–V460

132 132 133 133

V461–V463 V464–V469 V474–V479 V480–V485 V488 V492–V493 P494–P508 P509–P523 P524–P535 P536–P550 P551–P564 P565–P576 P577–P591 P592–P604 P605–P617

134 135 137 139 142 142 143 148 153 158 165 170 175 181 185

Fortbildung

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CONTENTS AUTHOR INDEX

Contents ∙ Inhalt ∙ (chronologisch) Oncol Res Treat 2016;39(suppl 3)

Abstract-No. Page-No.

Biologie der chronischen myeloischen Leukämie V619–V620 Zytopenien V625 Fortbildung Blutung – Gemeinsames Symposium mit der Gesellschaft für Thrombose- und Hämostaseforschung e. V. (GTH) V627–V629 Wissenschaftliches Symposium Patentrechtliche Probleme bei der personalisierten Therapie V630–V632 Immunphänotypisierung V634–V638 Systemmedizinische Ansätze in der Hämatologie / Onkologie V639–V642 Freier Vortrag Kolorektale Tumoren V643–V648 Fortbildung Keimzelltumoren V651 Expertenseminar Naturheilverfahren – Integrative Onkologie V654 Freier Vortrag Akute Leukämien V655–V660 Multiples Myelom klinisch V661–V666 Translationale Forschung V667–V672 Plenarsitzung Best Abstracts V673–V678 Wissenschaftliches Symposium Pathogenese und innovative Therapiekonzepte BCR-ABL-negativer myeloproliferativer Neoplasien V680–V682 Young investigators award V683–V688 Fortbildung Metastasiertes Mammakarzinom. Gemeinsames Symposium mit der Deutschen Gesellschaft für Senologie V693 Wissenschaftliches Symposium Allogene Stammzelltransplantation: aktuelle Konzepte V694 Fortbildung Mikroskopierkurs I V697–V698 Wissenschaftliches Symposium Survivorship, Rehabilitation – psychosoziale Belastungen V700–V701 Fortbildung Therapieoptimierung der akuten lymphatischen Leukämie V709–V710 Wissenschaftliches Symposium Liquid Biopsy V713 Freier Vortrag B-Zell-Lymphome, experimentell V715–V720 Chronische myeloische Leukämie – experimentell V723–V728 Multiples Myelom experimentell V729–V734 Wissenschaftliches Symposium Epigenetik myeloischer Neoplasien V737 Debatten MDS meets MPN V738–V739 Wissenschaftliches Symposium Biologie der chronischen lymphatischen Leukämie V741 Fortbildung Akute myeloische Leukämie V745–V747 Klug entscheiden V749 Wissenschaftliches Symposium Translationale Forschung V753–V754 Intensivmedizin nach allogener Stammzelltransplantation V755–V758 Freier Vortrag AYA – Heranwachsende und junge Erwachsene V759–V764 Wissenschaftliches Symposium Zell-basierte Immunmodulation – Gemeinsames Symposium mit der Deutschen Gesellschaft für Transfusionsmedizin und Immunhämatologie (DGTI) V766 Freier Vortrag Myeloproliferative Neoplasien – Klinik, Prognose V768–V773 Fortbildung Publish or perish? Meet the editor V774 Freier Vortrag Myelodysplastisches Syndrom – Prognose, Therapie, Monitoring V779–V784 Lymphome experimentell V785–V790 Palliativmedizin V791–V796 Debatten Risikoadaptierte Therapie des multiplen Myeloms V800–V801 Wissenschaftliches Symposium Biologie des myelodysplastischen Syndroms V802–V805 Biologie des Multiplen Myeloms V806–V808 Stammzellbiologie als Grundlage für therapeutische Interventionen V809–V810 Fortbildung Strahlung und Hämatologie V812–V815 Wissenschaftliches Symposium Betreuung und Rehabilitation chronischer Krebspatienten V817 Wissenschaftliches Symposium

Accessible online at: www.karger.com/ort

190 191 191 192 194 195 196 199 199 200 203 205 207 210 211 214 215 215 216 216 217 217 220 223 225 225 226 226 227 228 228 229

231 232 234 235 238 240 242 243 244 246 246 247

CONTENTS AUTHOR INDEX

Contents ∙ Inhalt ∙ (chronologisch) Oncol Res Treat 2016;39(suppl 3)

Abstract-No. Page-No.

Betreuung von Heranwachsenden und jungen Erwachsenen (AYA) Wissenschaftliches Symposium HPV-assoziierte Malignome Freier Vortrag Akute myeloische Leukämie – experimentell Tumor-/Zellbiologie Debatten MRD-Negativität: Ein klinisch relevantes Therapieziel bei der CLL? Posterdiskussion Akute lymphatische Leukämie, chronische lymphatische Leukämie Gastrointestinale Tumoren, Hepatozelluläres Karzinom, Pankreaskarzinom Nierentumoren, Prostatakarzinom Tumorbiologie, Zellbiologie 1 Stammzellmobilisierung; Zellbiologie 2 Neue Substanzen, sonstige Onkologie Heranwachsende und junge Erwachsene (AYA), Langzeitüberlebende, Patientensicherheit Supportivtherapie, Infektionen Kasuistiken – Infektionen und Onkologie Wissenschaftliches Symposium Biologie und Therapie der indolenten Lymphome Fortbildung Kompetenznetz – Akute und chronische Leukämie Freier Vortrag Ethik und Ökonomie Lungenkarzinome, Sarkome Kopf-/Hals-Tumoren Fortbildung Kompetenznetz Maligne Lymphome Freier Vortrag Allogene Stammzelltransplantation Immuntherapie experimentell Gastrointestinale Tumoren, Pankreaskarzinom Urogenitale Tumoren Plenarsitzung Zurück in die Zukunft Pflegetagung Patientenschulung und -beratung Neue Therapien – ärztliche und pflegerische Aspekte I Selbstmanagement Palliativpflege Spezialisierte onkologische pflegerische Versorgung Klangschalenmassage / Klangschalenmeditation Ekel und Scham Ergotherapie in der Palliativmedizin Fort- und Weiterbildung, Akademisierung, Zukunftsperspektiven Studententag Berufliche Perspektiven in Hämatologie & Onkologie

V820 V823–V825 V830–V835 V838–V843 V846 P847–P861

247 247 248 250 253 253

P862–P876 P877–P888 P889–P903 P904–P915 P916–P930

258 264 269 274 279

P931–P945 P946–P960 P961–P969 V970–V973 V975–V976 V979–V984 V985–V990 V995–V1000 V1005–V1008 V1010–V1015 V1016–V1021 V1022–V1027 V1028–V1033 V1034

285 291 297 301 302 302 304 307 309 310 313 316 318 321 322 322 322 323 323 324 325 325 325 326 326

Fortbildung

Promotionsstipendien

Author Index

Imprint

Accessible online at: www.karger.com/ort

329 350

CONTENTS AUTHOR INDEX

Contents ∙ Inhalt ∙ (nach Sitzung) Oncol Res Treat 2016;39(suppl 3)

Abstract-No. Page-No.

Eröffnungsveranstaltung V14 Best Abstracts V673–V678 Zurück in die Zukunft V1034 Wissenschaftliches Symposium Neue immuntherapeutische Ansätze nach allogener Stammzelltransplantation V75 ALL: Molekulare und zelluläre Grundlagen für zielgerichtete Therapien V77–V79 Neue Optionen beim Hodgkin-Lymphom V160 Kontroversen in der Therapie des kolorektalen Karzinoms V335 Wie teuer dürfen Arzneimittel sein? V370–V371 Therapeutische Targets bei AML – Vision und Wirklichkeit V458–V460 Wirkungen und Nebenwirkungen im Versorgungsalltag V488 Biologie der chronischen myeloischen Leukämie V619–V620 Zytopenien V625 Patentrechtliche Probleme bei der personalisierten Therapie V630–V632 Immunphänotypisierung V634–V638 Systemmedizinische Ansätze in der Hämatologie / Onkologie V639–V642 Pathogenese und innovative Therapiekonzepte BCR-ABL-negativer myeloproliferativer Neoplasien V680–V682 Young investigators award V683–V688 Allogene Stammzelltransplantation: aktuelle Konzepte V694 Survivorship, Rehabilitation – psychosoziale Belastungen V700–V701 Liquid Biopsy V713 Epigenetik myeloischer Neoplasien V737 Biologie der chronischen lymphatischen Leukämie V741 Translationale Forschung V753–V754 Intensivmedizin nach allogener Stammzelltransplantation V755–V758 Zell-basierte Immunmodulation – Gemeinsames Symposium mit der Deutschen Gesellschaft für Transfusionsmedizin und Immunhämatologie (DGTI) V766 Biologie des myelodysplastischen Syndroms V802–V805 Biologie des Multiplen Myeloms V806–V808 Stammzellbiologie als Grundlage für therapeutische Interventionen V809–V810 Betreuung und Rehabilitation chronischer Krebspatienten V817 HPV-assoziierte Malignome V823–V825 Biologie und Therapie der indolenten Lymphome V970–V973 Fortbildung Management der chronischen myeloischen Leukämie V15 Gynäkologische Tumoren V25 Palliativmedizin V27–V28 Kopf-Hals-Tumoren V35 Indolente Non-Hodgkin-Lymphome V63–V65 CUP-Syndrom V66–V67 Lebertumoren V69 Hirnmetastasen und ZNS-Lymphome V80–V82 Thrombophilie V85 Interprofessionelle Sitzung: Geriatrische Onkologie (für Ärzte und Pflegekräfte) V100–V101 Immuntherapie in der Hämatologie und Onkologie V110–V112 Mammakarzinom: (Neo)adjuvante Therapie V114 Plenarsitzung

Accessible online at: www.karger.com/ort

1 207 321 16 16 40 101 114 133 142 190 191 192 194 195 210 211 215 216 217 225 226 228 228

231 243 244 246 247 247 301 1 1 1 6 14 15 15 17 18 23 27 27

CONTENTS AUTHOR INDEX

Contents ∙ Inhalt ∙ (nach Sitzung) Oncol Res Treat 2016;39(suppl 3)

Abstract-No. Page-No.

ZNS-Tumoren Kontroverse Indikationen zur Stammzelltransplantation Lebensqualitäts-Assessment Eosinophilie, Mastozytose Interprofessionelle Sitzung: Therapiebegrenzung (für Ärzte und Pflegekräfte) Intensivmedizin und Hämatologie/Onkologie Standardisierte Leukämie-Diagnostik Myelodysplastisches Syndrom Chronische lymphatische Leukämie Sarkome 2016 – was gibt es Neues? Melanom Blut ist ein besonderer Saft – Transfusionsmedizin Nierenzellkarzinom Multiples Myelom Pankreaskarzinom Noch kurativ oder schon palliativ? Ethik in der Onkologie Schwangerschaft und Krebs Migrationsanämien Innovation durch Kooperation – Die Deutsche CML-Allianz Update Infektionen in der Hämatologie und Onkologie – 20 Jahre AGIHO Aggressive Non-Hodgkin-Lymphome Magenkarzinome Management myeloproliferativer Neoplasien Management des Hodgkin-Lymphoms Supportive Therapien: aktuelle Leitlinien in der onkologischen Praxis Blutung – Gemeinsames Symposium mit der Gesellschaft für Thrombose- und Hämostaseforschung e. V. (GTH) Keimzelltumoren Metastasiertes Mammakarzinom. Gemeinsames Symposium mit der Deutschen Gesellschaft für Senologie Mikroskopierkurs I Therapieoptimierung der akuten lymphatischen Leukämie Akute myeloische Leukämie Klug entscheiden Publish or perish? Meet the editor Strahlung und Hämatologie Betreuung von Heranwachsenden und jungen Erwachsenen (AYA) Kompetenznetz – Akute und chronische Leukämie Kompetenznetz Maligne Lymphome Expertenseminar Patienten mit ZNS-Lymphomen Diagnostische und therapeutische Fallstricke bei MPN-assoziierter Thrombophilie Naturheilverfahren – Integrative Onkologie Freier Vortrag Management hämatologischer Erkrankungen älterer Patienten Infektionsmanagement Rehabilitation

V117 V119–V122 V123–V126 V127–V130

27 28 29 30

V138–V140 V150–V151 V168–V169 V313–V316 V319 V321–V323 V327 V329 V336–V337 V359–V361 V367 V374–V377 V381 V396–V398 V414–V418

33 36 40 98 99 100 100 101 101 112 113 114 115 121 125

V421 V430–V432 V434–V435 V451–V453 V454

127 127 127 132 133

V461–V463

134

V627–V629 V651

191 199

V693 V697–V698 V709–V710 V745–V747 V749 V774 V812–V815 V820 V975–V976 V1005–V1008 V352

214 215 216 226 227 234 246 247 302 309 109

V444 V654 V29–V34 V42–V47 V48–V53

132 199 2 7 9

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CONTENTS AUTHOR INDEX

Contents ∙ Inhalt ∙ (nach Sitzung) Oncol Res Treat 2016;39(suppl 3)

Chronische lymphatische Leukämie – Biologie Zellbiologie Infektionen / supportive Therapie Mastozytose, myeloproliferative Neoplasien Nichtkleinzelliges Lungenkarzinom Chronische lymphatische Leukämie – Therapie Chronische myeloische Leukämie – Erstlinien- und Absetzstudien Versorgungsforschung Akute myeloische Leukämie – Therapie 1, Mutationsprofil Immuntherapie solider Tumoren Epigenetik Akute myeloische Leukämie – Therapie 2 Mammakarzinom Niedrigmaligne B-Zell-Lymphome – Klinik Myelodysplastisches Syndrom – experimentell Graft-versus-Host-Disease Immuntherapie von Leukämien und Lymphomen Nicht maligne Hämatologie (Minimale) Resterkrankung nach Leukämietherapie Kolorektale Tumoren Akute Leukämien Multiples Myelom klinisch Translationale Forschung B-Zell-Lymphome, experimentell Chronische myeloische Leukämie – experimentell Multiples Myelom experimentell AYA – Heranwachsende und junge Erwachsene Myeloproliferative Neoplasien – Klinik, Prognose Myelodysplastisches Syndrom – Prognose, Therapie, Monitoring Lymphome experimentell Palliativmedizin Akute myeloische Leukämie – experimentell Tumor-/Zellbiologie Ethik und Ökonomie Lungenkarzinome, Sarkome Kopf-/Hals-Tumoren Allogene Stammzelltransplantation Immuntherapie experimentell Gastrointestinale Tumoren, Pankreaskarzinom Urogenitale Tumoren Akute myeloische Leukämie Chronische myeloische Leukämie Myelodysplastisches Syndrom, sonstige Hämatologie Multiples Myelom 1 Lymphome Allogene Stammzelltransplantation 1 Immuntherapie Lungentumoren

Posterdiskussion

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Abstract-No. Page-No.

V56–V61 V86–V91 V92–V97 V103–V108 V131–V135 V141–V146 V153–V158 V170–V175 V178–V183 V339–V344 V345–V350 V353–V358 V384–V389 V390–V395 V401–V406 V437–V422 V464–V469 V474–V479 V480–V485 V643–V648 V655–V660 V661–V666 V667–V672 V715–V720 V723–V728 V729–V734 V759–V764 V768–V773 V779–V784 V785–V790 V791–V796 V830–V835 V838–V843 V979–V984 V985–V990 V995–V1000 V1010–V1015 V1016–V1021 V1022–V1027 V1028–V1033 P186–P200 P201–P215 P216–P226 P227–P241 P242–P256 P257–P271 P272–P285 P286–P300

11 18 20 24 31 34 37 41 44 102 106 109 116 118 122 128 135 137 139 196 200 203 205 217 220 223 229 232 235 238 240 248 250 302 304 307 310 313 316 318 46 52 59 64 70 76 82 87

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Contents ∙ Inhalt ∙ (nach Sitzung) Oncol Res Treat 2016;39(suppl 3)

Abstract-No. Page-No.

Kolorektale Karzinome Akute myeloische Leukämie, experimentell Myeloproliferative Neoplasien, Gerinnung Aggressive Non-Hodgkin-Lymphome Multiples Myelom 2 Allogene Stammzelltransplantation 2 Lungentumoren, Kopf-Hals-Tumoren Palliativmedizin, Integrative Onkologie Gynäkologische Tumoren, Mammakarzinom Kasuistiken – Hämatologie Akute lymphatische Leukämie, chronische lymphatische Leukämie Gastrointestinale Tumoren, Hepatozelluläres Karzinom, Pankreaskarzinom Nierentumoren, Prostatakarzinom Tumorbiologie, Zellbiologie 1 Stammzellmobilisierung; Zellbiologie 2 Neue Substanzen, sonstige Onkologie Heranwachsende und junge Erwachsene (AYA), Langzeitüberlebende, Patientensicherheit Supportivtherapie, Infektionen Kasuistiken – Infektionen und Onkologie Debatten CLL -Zukunft ohne Chemotherapie ? Ist die Zeit reif für eine therapiefreie Remission bei der CML? MDS meets MPN Risikoadaptierte Therapie des multiplen Myeloms MRD-Negativität: Ein klinisch relevantes Therapieziel bei der CLL? Pflegetagung Patientenschulung und -beratung Neue Therapien – ärztliche und pflegerische Aspekte I Selbstmanagement Palliativpflege Spezialisierte onkologische pflegerische Versorgung Fort- und Weiterbildung, Akademisierung, Zukunftsperspektiven Klangschalenmassage / Klangschalenmeditation Ekel und Scham Ergotherapie in der Palliativmedizin Studententag Berufliche Perspektiven in Hämatologie & Onkologie

P301–P312 P494–P508 P509–P523 P524–P535 P536–P550 P551–P564 P565–P576 P577–P591 P592–P604 P605–P617 P847–P861

93 143 148 153 158 165 170 175 181 185 253

P862–P876 P877–P888 P889–P903 P904–P915 P916–P930

258 264 269 274 279

P931–P945 P946–P960 P961–P969 V411 V492–V493 V738–V739 V800–V801 V846

285 291 297 125 142 225 242 253 322 322 322 323 323 325 324 325 325 326 326

Promotionsstipendien

Author Index

Imprint

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329 350

CONTENTS AUTHOR INDEX

Abstracts Oncol Res Treat 2016;39(suppl 3):1–350 DOI: 10.1159/000449050

Plenarsitzung

Eröffnungsveranstaltung V14

Nuclear waste disposal – what do we pass to our children? Stumpf T.1 Institut für Ressourcenökologie / HZDR, Dresden, Germany

become general recommendation in a well-defined patient population, the next step will be to think about how to increase patient numbers fulfilling these definitions. In this context, many factors have to be considered, e.g. optimizing first-line therapy, switching patients in MMR to other drugs to gain deeper MR. With the initial use of 2nd generation TKIs faster and deeper MR is achievable. The specific impact on TFR should be validated and current 1st line treatment strategies should be critically assessed. Further, the addition of other drugs, e.g. IFN or stem cell active drugs, may also increase the proportion of candidates for cessation attempts.

The commission of the German Parliament which deals with the law for the site selection will finish their work until this year. With the final report a new chapter in the field of searching and building a nuclear waste repository in Germany will be opened. Which kind of criteria are of importance for the site selection process? Where are the potential sites in Germany located? Are there any alternatives to a disposal in deep geological formations? All these questions will be discussed within the talk “Nuclear waste disposal – what do we pass to our children?”. Furthermore, the scientific background of the formation, conditioning and disposal of high radioactive waste will be illustrated.

Disclosure: Susanne Saußele: Advisory Role: Novartis, BMS, Pfizer, Ariad; Financing of Scientific Research: Novartis, BMS, Pfizer, Ariad; Expert Testimony: Novartis, BMS

Disclosure: No conflict of interest disclosed.

Letsch A.1

Fortbildung

Gynäkologische Tumoren V25

Immunotherapeutic approaches in gynecological cancers Med. Klinik m.S. Hämatologie, Onkologie und Tumorimmunologie, Charité Campus Benjamin Franklin, Berlin, Germany 1

Fortbildung

Management der chronischen myeloischen Leukämie V15

Therapy goals for first-line treatment in CML and implications for the doctor Saußele S.1 Medizinische Fakultät Mannheim der Universität Heidelberg, III. Med. Klinik, Mannheim, Germany 1

In CML, three tyrosine kinase inhibitors (TKI, dasatinib, imatinib, and nilotinib) are registered and recommended by expert groups for first-line treatment in CML. In order to choose optimal treatment for the individual patient with newly diagnosed CML different aspects have to be considered, e.g. impact of the TKI on overall survival (OS), progression-free survival (PFS), molecular response, side effects, and quality of life (QoL) as well as pre-existing comorbidities. First line studies comparing 2nd generation TKIs vs. imatinib (ENESTnd and DASISION study) demonstrated significant higher rates for all molecular remission levels like MMR (BCR-ABL (IS) level < 0.1%), MR4 (BCR-ABL (IS) < 0.01%), and MR4.5 (BCR-ABL (IS) < 0.0032%) and lower rates of PFS in patients randomized to the nilotinib and dasatinib arms compared with the imatinib arms; however, both studies failed to show a significant difference in OS. This could be due to the fact that patients often switch to a second- or third-line therapy, leading to a more difficult assessment of a benefit in OS. In addition, it was shown that comorbidities at diagnosis have major influence on the outcome and hence CML unrelated-deaths influence OS. Another hypothesis could be that the lower rate of PFS with the more potent TKIs cpuld be antagonized by a negative effect of these drugs in regard of adverse events; e.g. pulmonary toxicity with dasatinib and vascular toxicity with nilotinib. On the other hand, for the majority of patients, CML is now a chronic condition maintained by regular TKI therapy, so tolerability of the TKIs and their impact on Qol are of highest interest. In addition, the possibility to probably stop treatment (treatment-free remission (TFR)) moved in the focus since the publication of the STIM trial in 2010. If this option will

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The immune system is able to control cancer through various and dynamic interactions with cancer cells. Long-term remissions can be achieved by sustained immune responses against recurring cancer cells. The identification of immune checkpoint inhibitors, seems to be one of the most promising approaches of immunotherapy, currently known. The correlation between T cells in the tumor microenvironment and improved progression-free and overall survival in ovarian cancer, is only one factor indicating that immunotherapy could hold promise in gynecological cancers. The use of immune checkpoint inhibitors directed against programmed death receptors PD-1 and PD1-ligand and combinatorial approaches covering immunotherapeutics and conventional treatments will be discusses with focus on gynecological cancers. Furthermore the need for specific consideration of immune-related adverse effects, specific response criteria and potential biomarkers will be addressed. Although the clinical experience with novel immunotherapeutics remains limited in gynecologic cancer so far, early trials and clinical activity at least in specific subgroups of patients strongly supports the need for further investigation of immunotherapy approaches and their a potential role in the future treatment of gynecological cancers. Disclosure: Anne Letsch: Expert Testimony: Celgene, Novartis

Fortbildung

Palliativmedizin V27

Palliative Care for patients in clinical trials Simon S.T.1 Uniklinik Köln, Zentrum für Palliativmedizin, Köln, Germany

Palliative Care is part of high quality cancer care for patients with incurable cancer. National and international evidence-based guidelines state that palliative care should be integrated early after the diagnosis of incurable cancer. Palliative care should be offered regardless of whether cancer-spe-

CONTENTS AUTHOR INDEX

cific and disease modifying therapies are implemented. Clinical trials evaluate new and innovative approaches in order to optimize clinical care. Clinical trials are essential to improve both cancer care and palliative care. Patients in clinical trials should not be excluded from palliative care and have the same right to receive high quality cancer care including palliative care. Clinicans and researcher in oncology need to ensure that patients with incurable cancer in clinical trials have access to palliative care. Palliative care teams need to offer palliative care regardless of whether the patient take part in a clinical trial or not. Disclosure: Steffen Simon: Expert Testimony: Teva GmbH (Finanzierung einer IIT clinical trial); Otsuka GmbH (Finanzierung retrospektive Studie); Immaterial Conflict of Interests: Mitglie der Dt. Gesellschaft für Palliativmedizin und der Dt. Gesellschaft für Innere Medizin V28

When is it time to discontinue cancer treatment? Jahn-Kuch D.1 Medizinische Universität Graz, Klin. Abteilung für Onkologie/Universitäre Palliativmedizinische Einrichtung ; UKIM, Graz, Austria 1

The launch of new tumorspecific agents has significantly increased the life expectancy of patients suffering from advanced cancer during the past decades. Therefore a lot of studies show the increasing use of antineoplastic agents even in end-stage cancer patients with the intention to prolong life and to reduce cancer related symptoms regardless to the very small benefit in third-, forth or later therapy lines. Research data regarding the positive effects on patient survival or even more important quality of life of aggressive cancer treatment at the end of life are still scarce. The available publications demonstrate that up to a fifth of all patients with extremely advanced cancer are undergoing chemotherapy in the last month of life without apparant signs of benefit. It has been clearly demonstrated that antineoplastic therapy administered to patients at end stage disease produces a lot of adverse effects, leads to an increase in emergency department visits , increases days spent in hospital towards end of life, prevents patients from engaging in meaningful life review and preparing for death and is associated with a considerably underuse of hospice service. Studies document that even in patients with good performance status and advanced disease tumorspecific therapy can have a negative impact on quality of life. Patients of young age or with short history of metastatic disease are most likely to receive chemotherapy near the end of life. One of the main reasons for the overuse of tumorspecific therapy in this peculiar setting could be the overestimation of survival by medical oncologists. The prediction of a cancer patient’s life expectancy by physicians is accurate in only about 30-40% of cases. The decision to continue or to end antitumor-therapy is usually based on patient´s performance status, stage of disease, sensitivity of the tumor to antineoplastic therapies, blood tests, organ function and present symptoms. Nevertheless it is equally important for the process of decision making on further tumorspecific therapy to clearly address the limitations of antitumor therapy at end-stage cancer in time, to evaluate and be responsive to the patients wishes and to openly discuss end-of-life issues with the patient. This might help to avoid the application of useless treatments at the end of life. Disclosure: No conflict of interest disclosed.

Oncol Res Treat 2016;39(suppl 3):1–2

Freier Vortrag

Management hämatologischer Erkrankungen älterer Patienten V29

Hematopoietic Stem Cell Transplantation (HSCT) as compared to Consolidation Chemotherapy (CT) increases leukemia free survival in patients between 60 and 75 years with Acute Myelogenous Leukemia (AML) irrespective of the genetic risk: Report from the AML 2004 of the East German Study Group (OSHO) Niederwieser D.1, Al-Ali H.K.1, Krahl R.1, Kahl C.2, Wolf H.-H.3, Kreibich U.4, Vucinic V.1, Hähling D.5, Hegenbart U.6, Krämer A.6, Hirt C.7, Peter N.8, Opitz B.9, Florschütz A.10, Reifenrath K.11, Schulze A.12, Zojer N.13, Scholl S.14, Jakob C.15, Junghanss C.16, Pönisch W.1, Heyn S.1, Sayer H.G.12, Hochhaus A.14, Heinicke T.17, Fischer T.17, Dreger P.6, Maschmeyer G.18 University Hospital of Leipzig, Division of Hematology and Oncology, Leipzig, Germany, 2Hospital Magdeburg, Department of Hematology and Oncology, Magdeburg, Germany, 3University of Halle, Department of Hematology and Oncology, Halle, Germany, 4Heinrich-Braun Hospital Zwickau, Department of Hematology, Oncology and Palliative Care, Zwickau, Germany, 5Oncology Practice, Schwerin, Germany, 6University Hospital of Heidelberg, Department of Hematology, Oncology and Rheumatology, Heidelberg, Germany, 7University Hospital Greifswald, Clinic of Internal Medicine, Greifswald, Germany, 8CarlThiem-Hospital, Divison of Hematology and Oncology, Cottbus, Germany, 9 St. Elisabeth and St. Barbara Hospital Halle, Medical Clinic II, Halle, Germany, 10 Städtisches Klinikum Dessau, Department of Hematology and Oncology, Dessau, Germany, 11Hospital Zittau, Zittau, Germany, 12Helios Hospital Erfurt, Hematology, Oncology and Hemostaseology, Erfurt, Germany, 13 Wilhelminenspital Wien, Centre of Hematology and Oncology, Wien, Austria, 14 University Hospital Jena, Divison of Hematology and Oncology, Jena, Germany, 15Imperial College of London, London, United Kingdom, 16University Hospital Rostock, Clinic of Hematology, Oncology and Palliative Care, Rostock, Germany, 17University Hospital Magdeburg, Clinic of Hematology and Oncology, Magdeburg, Germany, 18Hospital Ernst von Bergmann, Department of Hematology, Oncology and Palliative Care, Potsdam, Germany 1

HSCT has been reported to be an option for elderly patients with AML. In this analysis the outcome was compared in regard to CT or HSCT and analyzed according to genetic risk groups defined by the ELN. By May 2015, 789 patients (60-75 a) of the AML 2004 study were identified as eligible. After induction with cytarabine (AraC; 1 g/m² i.v. bid d1, 3, 5, 7)/mitoxantrone (Mito; 10 mg/m² d1-3), 492 (62%) patients entered complete remission (CR) 1 with ranges from 79% for good risk (GR), 65% for intermediate (IM)-I, 59% for IM-II to 59% for high risk (HR) patients. A total of 355 patients were eligible either for CT with AraC (0.5 g/m2 i.v. bid d1, 3, 5)/Mito (10 mg/m² d1-2) (n = 205) or for matched HSCT (n = 119), if a donor was available. Most of the patients with HCT had unrelated (74.8%) donors and received low dose TBI (80.6%) as conditioning regimen. Median age was 68 and 66 a in the CT and HCT groups, respectively (p < 0.0005). There were no statistical significant differences regarding gender, AML type, NPM1 and FLT3 mutation status, but IM-II and HR cytogenetics were less frequent in the CT than in the HCT arm (p < 0.002). In addition, the interval from CR to CT was significantly shorter (43 d) than from CR to HCT (65 d; p < 0.0005]. Patients receiving matched HCT had superior LFS at 9 a than those receiving CT (25 ± 5% vs. 14 ± 3%, respectively; p < 0.001). As expected, relapse incidence (RI) was lower after HCT (42 ± 5%) than after CT (78 ± 3%; p < 0.0001) and non-relapse mortality (NRM) was higher in HCT (34 ± 5%) than in CT patients (8 ± 2%; p < 0.0001). Analyses of risk categories identified differences in LFS at 9 a between matched HCT and CT for IM-I (28 ± 8% vs. 16 ± 4%; p = 0.007), for IM-II (30 ± 10% vs. 4 ± 4%; p = 0.08) and for HR (12 ± 7% vs. 0 ± 0%; p < .05). The differences in LFS were due to decreased RI in matched HCT vs. CT for IM-I (35 ± 7% vs. 74 ± 4%; p < .0001), for IM-II (38 ± 10% vs. 96 ± 5%; p < .0001) and for HR (59 ± 11% vs. 96 ± 7%; p < .004). These effects outweigh the higher NRM in HCT vs CT for IM-I (37 ± 9% vs. 10 ± 3%; p = 0.0005), for IM-II

Abstracts

CONTENTS AUTHOR INDEX

(33 ± 10% vs. 0%; p = 0.003) and for HR (29 ± 10% vs. 4 ± 4%; p = 0.11). Independent prognostic factors for LFS were genetic risk group and HCT (p < 0.0005), for OS genetic risk group (p < 0.01), for RI were genetic risk group and CT (p < 0.0005) and for NRM HCT (p < 0.005). HCT from related/unrelated donors improves LFS in elderly patients with AML. This improvement is noted in all genetic risk groups from IM-I to HR. Disclosure: No conflict of interest disclosed. V30

Outcome of patients ≥ 65 years of age suffering myelofibrosis and treated with RIC allogeneic hematopoietic stem cell transplantation Christopeit M.1, Alchalby H.1, Zabelina T.1, Zeck G.1, Ayuk F.A.1, Wolschke C.1, Kröger N.1 Universitätsklinikum Eppendorf, Klinik für Stammzelltransplantation, Hamburg, Germany 1

Introduction: Median age of patients with myelofibrosis is 65 years at diagnosis. Frequently, patients of higher age are considered not fit for allogeneic transplantation. We hypothesized that RIC plus allogeneic transplantation is feasible for this population of patients and assessed outcomes of 53 patients ≥65 years with myelofibrosis. Patients and methods: Patients’ (22 female, 42%) median age at transplantation was 68 (65-75) years. Primary Myelofibrosis (PMF) had been diagnosed in 37 patients (70%), the disease was of post-ET/-PV origin in 16 patients (30%). At diagnosis, 24 patients showed normal karyotype (45%). Bone marrow fibrosis scored 1° in 3 patients (6%), 2° in 7 patients (13%), and 3° in 34 patients (64%). DIPSS was intermediate-1 in 3 patients (6%), intermediate-2 in 21 patients (40%), and high in 22 patients (42%). CMV IgG pairing between donor/ patient was negative/ negative in 17 patients (32%), positive/ positive in 26 patients (49%), positive/ negative in 4 patients (8%), and negative/ positive in 6 patients (11%). Intravenous Busulfan (10 doses of 0.8 mg/kg BW) and Fludarabin (6 doses of 30 mg/m²), combined with Anti-Lymphocyte Globulin at 30-90 mg/kg, was used for conditioning in 33 patients (62%), 17 patients (32%) received an intensified RIC scheme consisting of sequential FLAMSA-chemotherapy and Busulfan/ Fludarabine RIC, combined with 30-90 mg/kg Anti-Lymphocyte Globulin. The median of transplanted PBSC was 6.3×106 (1.5-16.1×106)/kgBW. A matched related donor was present for 7 patients (13%), matched unrelated for 27 patients (51%), 18 patients (34%) received their transplant from a mismatched unrelated donor, 1 patient from a haploidentical donor. Immunosuppression mainly (85%) consisted of cyclosporine and mycophenolic acid. Results: Leucocyte engraftment occurred in 49 patients (92%), 4 patients (8%) experienced primary graft failure. Median time to leucocyte engraftment was 13 (7-34) days, median time to platelet engraftment 20.5 (5-293) days. Acute GVHD grades 2-4 was present in 22 patients (42%), chronic GVHD in 19 patients (36%). Cumulative incidence (CI) of relapse was 21 ± 6% after 3 years. CI of non-relapse mortality (NRM) was 28 ± 6% after 1 year. OS at 3 years was 46 ± 8%. Conclusion: Allogeneic stem cell transplantation of patients with myelofibrosis is feasible even at and above 65 years of age. Further research will have to focus on further reducing NRM in this elderly population.

V31

Prevalence and dynamics of leukemia-associated mutations in elderly individuals without hematologic disorders Ernst T.1, Rinke J.1, Müller V.1, Waldau A.1, Midic D.1, Pester F.2, Landschulze J.2, Rudolph L.3, Hochhaus A.1 Klinik für Innere Medizin II, Universitätsklinikum Jena, Abteilung Hämatologie und Internistische Onkologie, Jena, Germany, 2HausArztZentrum, Kahla, Germany, 3Leibniz Institute for Age Research, Fritz Lipmann Institute e.V. (FLI), Jena, Germany 1

Introduction: Clonal hematopoiesis is frequently observed in elderly people and may represent a premalignant condition. Evidence suggests that genetic alterations found in patients with myeloid malignancies may also be causative for induction of clonal hematopoiesis in healthy elderly. Given these circumstances, discrimination between age-associated and leukemia-associated mutations is of major clinical importance. Methods: In order to investigate the prevalence and dynamics of genetic alterations among elderly individuals without hematologic/oncologic disorders, we genotyped a cohort of 50 elderly individuals (29 women; median age 84 years; range 80-90 years) for a panel of 30 commonly mutated leukemia-associated genes by targeted deep next-generation sequencing (NGS). Buccal cells were analyzed to confirm the somatic origin of mutations. Results: A total of 16 somatic mutations in leukemia-associated genes were identified in 13 of 50 (26%) elderly individuals. One subject presented with two, another subject with three different mutations. Ten of 16 mutations (63%) affected epigenetic modifier genes (DNMT3A, n = 8; TET2, n = 1; IDH2, n = 1). Four somatic mutations affected genes involved in the RNA splicing machinery (SRSF2, n = 2; SF3B1, n = 1; U2AF1, n = 1). Mutations in TP53 and NRAS were identified in two individuals. All but one mutation were missense mutations with cytosine to thymine transitions being the most common base pair change (n = 7). Mutations occurred at low variant allele frequencies (VAF) with a median of 11.7% (range: 1.0% to 30.7%) indicating that mutations were presented in only a subset of blood cells. Mutation kinetics remained virtually stable over a follow-up observation of two years as measured by targeted NGS (median VAF 13.1%, range 1.0% to 35.3%). Conclusions: These findings indicate that the appearance of low-level clones characterized by mutations in epigenetic regulator genes and the RNA splicing machinery is a common age-associated phenomenon which may represent a premalignant condition in the development of hematologic cancers and may also predispose to other aging associated disorders. The question why these subclones are selected in the elderly but apparently enter a stage of clonal size stability without further selection in healthy elderly in contrast to patients with myeloid disorders is of particular interest and needs further investigation. Disclosure: No conflict of interest disclosed.

Disclosure: No conflict of interest disclosed.

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

CONTENTS AUTHOR INDEX

V32

V33

The distinct genetic and epigenetic landscape of elderly AML: Data of the SAL registry

Evaluation of the international prognostic index for chronic lymphocytic leukemia (CLL-IPI) in elderly patients with comorbidities: Analysis of the CLL11 study population

Baldus C.D.1,2, Silva P.1, Neumann M.1,2, Vosberg S.3,4,5, Schlee C.1, Isaakidis K.1, Schroeder M.P.1, Ortiz Tanchez J.1, Fransecky L.R.1, Hartung T.1, Türkmen S.6, Graf A.3, Krebs S.3, Blum H.3, Thiede C.7,8, Ehninger G.7,8, Serve H.9,10, Berdel W.11, Greif P.A.4,5,12, Röllig C.7,8, Studienallianz Leukämie (SAL) Charite, Campus Benjamin Franklin, Hematology, Oncology, Berlin, Germany, DKTK, Berlin, Germany, 3Gene Center LMU München, München, Germany, 4 DKTK, München, Germany, 5Medizinische Klinik III, Klinikum der Universität München, München, Germany, 6Labor Berlin Charité Vivantes GmbH, Berlin, Germany, 7Medizinische Klinik und Poliklinik I, Universitätsklinikum Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany, 8DKTK, Dresden, Germany, 9Medizinische Klinik II, Universitätsklinikum Frankfurt am Main, Frankfurt, Germany, 10DKTK, Frankfurt, Germany, 11Medizinische Klinik, Universitätsklinikum, Münster, Germany, 12German Cancer Research Center (DKFZ), Heidelberg, Germany 1 2

Despite advances in the characterization of molecular alterations in younger AML patients, comprehensive studies in elderly AML are lacking to uncover its distinct molecular alterations. In this project, we investigated genetic and epigenetic modifications to unravel the specific molecular background of this unfavourable disease. To characterize elderly AML, we studied bone marrow samples of 93 AML patients enrolled on the Studienallianz Leukämie (SAL) registry. We selected a cohort of patients 65 to 90 years of age (median 72 years). To capture a broad spectrum of alterations we performed target enrichment of 555 candidate genes and sequenced them on an Illumina HiSeq 1500. We further investigated the DNA methylation profiles assessed them with an Infinium® HumanMethylation450 BeadChip on 79 available DNA samples (from the 93 samples). Overall, 814 molecular alterations were detected in 281 of 555 genes with a median of 7 genes mutated per patient (range, 1 to 23). Particularly high mutation frequencies were detected in DNMT3A 33%, TET2 24%, SRSF2 23%, ASXL1 21%, RUNX1 18%, IDH1 17%, NPM1 15%, IDH2 and BCOR 10%. We observed a high frequency of mutations in epigenetic regulators, which affected 85% (79/93) of patients and also found frequent alterations in splicing factors (SRSF2, U2AF1, SF3B1, ZRSR2, DDX5) affecting 38% (35/93) of patients. Notably, 17% of elderly AML patients had mutations in the DNA repair genes (TP53, NBN, ATM, FANCA, FANCC), which were associated with an inferior median overall survival (OS) of only 4 months, compared to 16 months for patients without alterations in these DNA repair genes (p < 0.001). Interestingly, mutations in DNA repair proteins predicted poor OS independently of the TP53 mutational status (p = 0.007). The unsupervised analyses of DNA methylation revealed a profile of hypermethylation specific to AML samples with IDH1/2 mutations. Samples defined by a triple mutation pattern (DNMT3A, NPM1, FLT3) presented with a markedly distinct profile, largely characterized by hypomethylation of specific regions. In conclusion, elderly AML harbours a high frequency of molecular alterations in epigenetic regulators, spliceosome components and DNA repair factors, the latter being associated with poor prognosis. This molecular categorization of elderly AML underscored a distinct biology and the need for specific molecularly driven therapeutic approaches.

Bahlo J.1, Goede V.1,2, Kutsch N.1, Fischer K.1, Fink A.-M.1, Stilgenbauer S.3, Bergmann M.4, Eichhorst B.1, Hallek M.1,5 Uniklinik Köln, Deutsche CLL Studiengruppe, Klinik I für Innere Medizin, Centrum für Integrierte Onkologie Köln-Bonn, Köln, Germany, 2St. Marien Hospital, Klinik für Altersmedizin, Köln, Germany, 3Uniklinik Ulm, Klinik für Innere Medizin III, Ulm, Germany, 4Klinikum München-Schwabing, Klinik für Hämatologie, Onkologie, Immunologie, Palliativmedizin, Infektiologie & Tropenmedizin, München, Germany, 5Exzellenzcluster Cellular Stress Responses in Aging-Associated Diseases (CECAD), Köln, Germany 1

Introduction: CLL-IPI is a validated tool for prognostication of overall survival (OS) in chronic lymphocytic leukemia using age, stage, β2-microglobulin, 17p deletion/TP53 mutation, and IGHV mutational status as weighted factors to stratify patients for low, intermediate, high, or very high risk of death. To date, validation studies of CLL-IPI have been restricted to patient populations with only moderately advanced age and little comorbidity. We here evaluated CLL-IPI in a large sample of elderly patients with comorbidities who represent the majority of patients with CLL outside of clinical trials. Methods: CLL-IPI was analyzed in the CLL11 study which enrolled 781 patients with previously untreated CLL and increased comorbidity for treatment with obinutuzumab (GA101) plus chlorambucil (G-Clb), rituximab plus chlorambucil (R-Clb), or chlorambucil alone (Clb). Patients with all five CLL-IPI factors available were stratified into CLL-IPI risk groups. Kaplan-Meier methodology was used to estimate OS for low, intermediate, high, and very high risk. Log-rank test and Cox regression were used to compare OS among CLL-IPI risk groups. Results: Among 781 patients enrolled in the CLL11 study, 691 patients were evaluable in this analysis while 90 subjects had to be excluded due to missing information for β2-microglobulin, 17p deletion/TP53 mutation, or IGHV mutational status. Of the 691 subjects, 299 were treated with G-Clb, 294 with R-Clb, and 98 with Clb. Median age, cumulative illness rating scale (CIRS), and ECOG performance status were 74 years, 8 and 1, respectively. Median observation time was 41.8 months. Stratification according to CLL-IPI was as follows: 62 (9%) low risk, 206 (30%) intermediate risk, 361 (52%) high risk, 62 (9%) very high risk. In a pooled analysis of all 691 evaluable patients, OS was significantly different across CLL-IPI risk groups (p < 0.001, see figure), with statistically satisfying values regarding both discrimination and calibration. Conclusions: This is the first validation study of CLL-IPI in elderly patients with previously untreated CLL and comorbidities. Results suggest good performance of the CLL-IPI in this patient population.

Disclosure: No conflict of interest disclosed.

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Fig. 1. CLL11_CLL-IPI. Disclosure: Jasmin Bahlo: Financing of Scientific Research: Roche Michael Hallek: Advisory Role: Roche; Financing of Scientific Research: Roche V34

International, randomized phase 3 study results: Ibrutinib versus Chlorambucil in patients 65 years and older with treatment-naïve CLL (RESONATE-2™) Burger J.1,2, Tedeschi A.3, Barr P.M.4, Robak T.5, Owen C.6, Ghia P.7, Bairey O.8, Hillmen P.9, Bartlett N.L.10, Li J.11, Simpson D.12, Grosicki S.13, Devereux S.14, Mccarthy H.15, Coutre S.16, Quach H.17, Gaidano G.18, Maslyak Z.19, Stevens D.A.20, Janssens A.21, Offner F.22, Mayer J.23, O’Dwyer M.24, Hellmann A.25, Schuh A.26, Siddiqi T.27, Polliack A.28, Tam C.S.29, Suri D.30, Cheng M.30, Clow F.30, Styles L.30, James D.F.30, Kipps T.J.31, for the RESONATE-2 Investigators The University of Texas MD Anderson Cancer Center, Houston, United States, Albert-Ludwigs University, School of Medicine, Freiburg, Germany, 3Azienda Ospedaliera Niguarda Cà Granda, Milano, Italy, 4Wilmot Cancer Institute, University of Rochester, Rochester, United States, 5Medical University of Lodz and Copernicus Memorial Hospital, Lodz, Poland, 6Tom Baker Cancer Centre, Calgary, Canada, 7Università Vita-Salute San Raffaele and IRCCS Istituto Scientifico San Raffaele, Milano, Italy, 8Rabin Medical Center, Beilinson Hospital and Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel, 9The Leeds Teaching Hospitals, St. James Institute of Oncology, Leeds, United Kingdom, 10Washington University School of Medicine, St. Louis, United States, 11Jiangsu Province Hospital, Nanjing, China, 12North Shore Hospital, Auckland, New Zealand, 13Department of Cancer Prevention, School of Public Health, Medical University of Silesia, Katowice, Poland, 14Kings College Hospital, London, United Kingdom, 15Royal Bournemouth Hospital, Bournemouth, United Kingdom, 16Stanford University School of Medicine, Stanford, United States, 17St. Vincent’s Hospital, University of Melbourne, Melbourne, Australia, 18Department of Translational Medicine, Amedeo Avogadro University of Eastern Piedmont, Novara, Italy, 19Institute of Blood Pathology and Transfusion Medicine of NAMS of Ukraine, Lviv, Ukraine, 20 Norton Cancer Institute, Louisville, United States, 21University Hospital Leuven, Leuven, Belgium, 22University Hospital Gent, Gent, Belgium, 23Fakultni Nemocnice Brno, Brno, Czech Republic, 24University College Hospital Galway, Galway, Ireland, 25 Department of Hematology, University Clinical Center of Medical University of Gdańsk, Gdańsk, Poland, 26University of Oxford, Oxford, United Kingdom, 27City of Hope National Medical Center, Duarte, United States, 28Hadassah University Hospital, Hebrew University Medical School, Jerusalem, Israel, 29Peter MacCallum Cancer Centre and St. Vincent’s Hospital, Melbourne, Australia, 30Pharmacyclics LLC, an AbbVie Company, Sunnyvale, United States, 31University of California San Diego, Moores Cancer Center, San Diego, United States 1 2

Introduction: Chronic lymphocytic leukemia (CLL) primarily affects older patients (pts) with medical comorbidities. Alkylating agents, such as chlorambucil (clb), are commonly used in these pts, but novel therapies are needed. Ibrutinib (ibr), a first-in-class, once-daily inhibitor of Bruton’s tyrosine

Abstracts

kinase, is approved in the EU for pts with CLL after ≥1 prior therapy and for pts with del17p CLL and/or TP53 mutation unsuitable for chemoimmunotherapy. Ibr is also indicated by the US FDA for the treatment of pts with CLL and for CLL with deletion 17p. Ibr has shown high activity in treatment-naïve (TN) pts age ≥65 y. This randomized open-label phase 3 trial evaluated efficacy and safety of single-agent ibr vs clb in TN older pts with CLL. Methods: Pts age ≥65 y with TN CLL were randomized 1:1 to receive 420 mg ibr daily until progression or clb 0.5 mg/kg (max 0.8 mg/kg) on d1 and 15 of a 28-d cycle (≥12 cycles). Primary endpoint was independent review committee (IRC)-assessed PFS per iwCLL 2008 criteria, with 2012 clarification for treatment-related lymphocytosis. Secondary endpoints were OS, ORR, rates of hematologic improvements, and safety. Results: 269 pts enrolled; median age 73 y. Baseline characteristics were balanced between arms. For clb-treated pts, 40% completed 12 cycles of therapy (mean dose 0.6 mg/kg). At median follow-up (18.4 mo), ibr significantly prolonged IRC-assessed PFS vs clb (median not reached vs 18.9 mo, P < 0.0001). Investigator-assessed 18-month PFS was 93.9% vs 44.8% for ibr vs clb (P < 0.0001). Ibr also significantly prolonged OS vs clb (P = 0.0010). Three deaths occurred on the ibr arm vs 17 deaths on the clb arm. IRC-assessed ORR was 86.0% with ibr vs 35.3% with clb; investigator-assessed ORR was 90.4% vs 35.3%. Rates of sustained hematologic improvement were significantly higher for ibr vs clb (84% vs 45%, P < 0.0001, anemia; 77% vs 43%, P = 0.0054, thrombocytopenia). Median duration of treatment was 17.4 mo (ibr) and 7.1 mo (clb). The most frequent (≥20% of pts) adverse events with ibr were diarrhea, fatigue, cough, and nausea and for clb were nausea, fatigue, neutropenia, anemia, and vomiting. Major hemorrhage occurred in 4% with ibr over median follow-up of 1.5 years and in 2% with clb. At study closure, 87% of ibr pts continue to receive ibr. Conclusions: Single-agent ibr was superior to clb for PFS, OS, ORR, and hematologic improvements in TN older CLL pts, with an 84% reduction in risk of death and an acceptable safety profile. Disclosure: Jan Burger: Advisory Role: Janssen, Boehringer Ingelheim, Portola; Expert Testimony: Pharmacyclics, Gilead; Other Financial Relationships: Travel, Accommodations, Expenses: Roche, Janssen Thomas Kipps: Advisory Role: AbbVie, Genentech, Gilead; Expert Testimony: AbbVie, Genentech, Pharmacyclics

Fortbildung

Kopf-Hals-Tumoren V35

Squamous cell carcinoma of head and neck: surgical and adjuvant treatment concepts in 2016 Dietz A.1 HNO-Universitätsklinik, Leipzig, Germany

In this update presentation, the current major considerations for the primary surgical treatment of squamous cell carcinoma of the head and neck region are presented and discussed. The reader will be introduced in less detail of surgica techniques than in the operational conceptual background of currently recommended treatment concepts. The Europe 5 year survival rate of squamous cell carcinoma of the head and neck region (HNSCC) is currently at 42%. Especially in the last 3 years, various guidelines have been established based on limited evidence for standardization of therapeutic concepts in HNSCC. If functional operability is possible In Europe It is found predominantly that primary surgical approaches are preferred. Postoperative adjuvant therapy is standardized due to clear indications based on defined risk situations. Indications are related to assessment of surgical margins, cervical lymph node metastases and extracapsular tumor growth. Ablative surgical procedures are competitive with so-called organ preservation programs which are currently addressed in clinical trials since many open questions regarding late functional outcome are still under discussion. Disclosure: Andreas Dietz: Advisory Role: Merck Serono, Astra Zeneka, MSD Merck, BSM,

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Freier Vortrag

V43

Infektionsmanagement

Parainfluenza in hematologic patients – impact of prolonged viral shedding and nosocomial infection

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Lehners N.1, Puthenparambil J.1, Schiller M.1, Ho A.D.1, Schnitzler P.2, Egerer G.1

Invasive mucormycosis in patients with hematological diseases identified in the global FungiScope™ registry Seidel D.1, Duran Graeff L.1, Vehreschild M.J.G.T.1, Liss B.1, Köhler P.1, Müller F.1, Wisplinghoff H.2, Vehreschild J.1, Cornely O.1 Uniklinik Köln, Klinik I für Innere Medizin, Cologne, Germany, 2Uniklinik Köln, Institut für Mikrobiologie, Cologne, Germany 1

Background: Invasive fungal diseases (IFD) are a frequent complication in hematological patients. Hematological malignancies and their treatment are main risk factors for IFD. Less frequent IFD, such as invasive mucormycoses (IM), are increasing worldwide and are associated with mortality up to 100%. Efficient diagnostic and treatment approaches for IM are not known or validated yet. Thus, it is urgent to better understand clinical presentation and management of IM in hematological patients to eventually improve patient outcome. Methods: Clinical data on IM were collected in FungiScope™, an international web-based retrospective registry. Cases with cultural, histological or molecular evidence of IM are enrolled. Data collected include demographics, underlying conditions and their treatment, clinical signs and symptoms, sites of infection, diagnostic tests, antifungal treatment and outcome. Clinical isolates are collected for centralized identification, molecular analyses and exchange between collaborators. Results: To date, 158 cases of IM with an underlying hematologic disease (HD) were captured in the FungiScope database. AML, ALL, Non-Hodgkin’s Lymphoma and CLL were the most frequent malignancies (46.8%, 19%, 7%, and 5.7%, respectively). Treatment of HD was chemotherapy in 75%, for 70% of these it was the primary course. Prolonged neutropenia (>10 days) was reported in 51% of cases. The majority of patients received antifungal agents (79%; median 42 days, range 1-733 days) and 37% underwent surgical debridement. Antifungals known to be active against Mucorales (amphotericin B, isavuconazole, itraconazole, posaconazole) were administered in 86% of cases, in 23% of those amphotericin B was used as a single agent. Median followup time was 46 days (range 0-1394 days). Favourable response (complete or partial response or stable disease) resulted in 42% of cases. Overall mortality was 65% and death due to IM was reported for 78% of cases. For patients who were not treated for IM, mortality exceeded 90%. Depending on the underlying disease overall mortality ranged from 61% for ALL and AML to 100% for CLL. Conclusion: The FungiScope registry is a feasible approach to collect data on a relevant amount of rare cases of IM. IM remains a life threatening disease in hematological patients. Despite aggressive antifungal treatment strategies outcome remains poor. More effective treatment strategies are urgently needed to improve patient outcome.

Universitätsklinikum Heidelberg, Abteilung für Hämatologie, Heidelberg, Germany, 2Universitätsklinikum Heidelberg, Zentrum für Infektiologie, Heidelberg, Germany 1

Introduction: Parainfluenza virus (PIV) is often cause of self-limiting upper respiratory tract infection (URTI), but can result in severe and even life-threatening lower respiratory tract infection (LRTI) in the immunocompromised host. In contrast to seasonal influenza, PIV is transmitted all year round. Here, we describe outcome and risk factors of PIV infection in hematologic patients and report on prolonged viral shedding in this particular population. Methods: Clinical characteristics and outcome of hematologic patients with documented PIV infection treated at our institution from July 2013 to June 2015 were retrospectively evaluated. In patients with available consecutive tests for PIV, duration of viral shedding was assessed. Results: 60 patients were identified, 47 with PIV 1/3, 13 with PIV 2/4. Median age was 59 years [range 26-76], 38 patients were male. 41 patients had received a stem cell transplantation (20 allogeneic, 17 autologous, 4 both). Nosocomial infection, defined as diagnosis of PIV infection ≥1 week after hospital admission, was apparent in 23 patients. In regard to outcome, 36 patients had URTI only, 24 (40%) developed a LRTI. Of the latter, 3 patients (13%) were transferred to the ICU and subsequently died. Neither type of PIV nor underlying hematologic disease had a significant impact on outcome. Leukopenia as well as nosocomial infection were significantly associated with LRTI (p = 0.02 and p = 0.01, resp.), a trend was seen for status post allogeneic transplantation (p = 0.06). Duration of viral shedding could be evaluated in 23 patients. Median duration of viral shedding was 14 days, but shedding of up to 79 days was observed. A significant association between LRTI and prolonged shedding for >20 days was observed (p = 0.01). Conclusion: PIV infection can cause significant morbidity in hematologic patients. Nosocomial infection was frequently present and was associated with a higher rate of LRTI in our patient population. Prolonged viral shedding was observed which might facilitate nosocomial spread of PIV and should be taken into account in infection control management. Disclosure: No conflict of interest disclosed.

Disclosure: Danila Seidel: No conflict of interest disclosed. Oliver Cornely: Advisory Role: Amplyx, Anacor, Astellas, Basilea, Cidara, Da Volterra, Daiichi Sankyo, F2G, Genentech/Roche, Gilead, Matinas, MedPace, Merck/ MSD, Merck Serono, Pfizer, Sanofi Pasteur, Scynexis, Seres, Summit, Vical, Vifor; Financing of Scientific Research: Astellas, Basilea, Gilead, Merck/MSD, and Pfizer; Expert Testimony: 3M, Actelion, Astellas, AstraZeneca, Basilea, Bayer, Celgene, Cubist/Optimer, Duke University (NIH UM1AI104681), Genzyme, Gilead, GSK, Leeds University, Merck/MSD, Miltenyi, NanoMR, Pfizer, Quintiles, Scynexis, Viropharma; Other Financial Relationships: supported by the German Federal Ministry of Research and Education

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Prophylactic exchange of central venous catheters (CVC) for prevention of CVC-related bloodstream infections (CRBSI) in patients with haematological malignancies: indirect evidence from pooled data of the SECRECY registry and the COAT study Schalk E.1, Biehl L.M.2,3, Färber J.4, Huth A.2, Panse J.5, Krämer C.5, Hentrich M.6, Engelhardt M.7, Schäfer-Eckart K.8, Kofla G.9, Kiehl M.10, Wendtner C.-M.11, Karthaus M.12, Cornely O.A.2,13,14, Fischer T.1, Vehreschild M.J.G.T.2,3 Otto-von-Guericke University Magdeburg, Medical Centre, Department of Haematology and Oncology, Magdeburg, Germany, 2University Hospital of Cologne, Department I of Internal Medicine, Cologne, Germany, 3German Centre for Infection Research (DZIF), Site Bonn/Cologne, Cologne, Germany, 4 Otto-von-Guericke University Magdeburg, Medical Centre, Department of Medical Microbiology, Infection Control and Prevention, Magdeburg, Germany, 5 RWTH Aachen University Hospital, Department of Haematology, Oncology, Haemostaseology, and Stem Cell Transplantation, Aachen, Germany, 6Red Cross Hospital Munich, Department of Medicine III, Munich, Germany, 7Medical Centre – University of Freiburg, Department of Medicine I, Haematology, Oncology and Stem Cell Transplantation, Freiburg, Germany, 8Klinikum Nuernberg, Medical Clinic V, Haematology and Oncology, Nuernberg, Germany, 9Charité – University Medicine Berlin, Department of Medicine, Division of Oncology/Haematology, Berlin, Germany, 10Clinical Centre Frankfurt/Oder, Medical Clinic I, Haematology and Medical Oncology, Frankfurt/Oder, Germany, 11Klinikum Schwabing, Department of Haematology, Oncology, Immunology, Palliative Care, Infectious Diseases and Tropical Medicine, Munich, Germany, 12Klinikum Neuperlach, Department of Haematology and Oncology, Munich, Germany, 13University of Cologne, Centre for Integrated Oncology CIO Köln/Bonn, Cologne, Germany, 14 University of Cologne, Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), Cologne, Germany 1

Introduction: CVC exchange in cancer pts. is not general recommended to prevent CRBSI, but it is sometimes still practice and the optimal time point is unclear. However, this recommendation is based on studies that were underpowered for haematological pts. Therefore, we aimed to verify this question in a larger cohort of pts. with haematological malignancies. Methods: Prospective, multicentre analysis of CRBSI due to short-term CVC (≥1 day in situ) in pts. with haematological malignancies. Pooled data of the SECRECY registry (DRKS00006551) and the COAT study (NCT01544686) were used. For CRBSI definition the 2012 AGIHO/ DGHO criteria were used; only definite and probable CRBSI were considered. Length of catheterization (LOC) was used to determine a cut-off for CRBSI risk using area under the receiver operating characteristic curve (AUC); AUC< 0.50 was considered to be of none predictive value. Also, CRBSI risk was calculated comparing LOC for quartiles 1 to 4 (Q1 to Q4). Results: Altogether, 1083 CVC (median age = 59 years [range 18-86], 59.5% men) with 18,327 CVC days (median LOC = 17 days [range 1-60]) were analysed in 857 pts. in 10 centres. Underlying diseases were acute leukaemia (46.8%), multiple myeloma (26.4%) or lymphoma (19.0%). Most of the CVC were inserted in the jugular vein (n = 699, 64.5%). The CRBSI rate was 11.6% (n = 126). The CRBSI incidence and the cumulative incidence were 6.9/1000 CVC days and 30.3%, respectively. The CRBSI onset was in median on day 13 (range 2-40). The predominant pathogenic agents were coagulase-negative staphylococci (n = 95, 75.4%). The LOC quartiles were day 1-12 for Q1, day 13-17 for Q2, day 18-23 for Q3 and day 24-60 for Q4, respectively. The CRBSI risk was higher for LOC Q2 vs. Q1 (hazard ratio [HR]=3.62 [95%CI 1.94-6.74]; p < 0.001), for Q3 vs. Q2 (HR = 4.92 [95%CI 2.57-9.42]; p < 0.001) and for Q4 vs. Q3 (HR = 2.36 [95%CI 1.23-4.52]; p = 0.01). For the LOC cut-off for CRBSI prediction an AUC of 0.45 (95%CI 0.40-0.49) was calculated. There was no higher CRBSI risk for LOC more than the median LOC (Q3-Q4 vs. Q1-Q2) (relative risk [RR]=1.02 [95%CI 0.73-1.43]; p = 0.89), but for LOC more than median CRBSI onset (equivalent to Q2-Q4 vs. Q1) (RR = 1.89 [95%CI 1.25-2.86]); p = 0.003). Conclusions: The CRBSI risk is increasing with LOC in pts. with haematological malignancies. However, there is no LOC cut-off for prediction CRBSI. Therefore, within the first 2 weeks of LOC no recommendation can be given for CVC exchange to prevent CRBSI.

Abstracts

Disclosure: Enrico Schalk: No conflict of interest disclosed. Maria Vehreschild: Advisory Role: MJGT has been a consultant to Astellas Pharma, Berlin Chemie, Merck/MSD and DaVolterra.; Financing of Scientific Research: MJGTV has served at the speakers’ bureau of Pfizer, Merck/MSD, Gilead Sciences and Astellas Pharma.; Expert Testimony: MJGT received research funding from 3M, DaVolterra, Gilead Sciences and Astellas Pharma. V45

Epidemiology and outcomes of infections during neutropenia after 166 allogeneic stem cell transplantations in one year at one center Samek M.1, Wolschke C.1, Langebrake C.1, Adjallé R.1, Ayuk F.A.1, Kröger N.1, Christopeit M.1 Universitätsklinikum Eppendorf, Klinik für Stammzelltransplantation, Hamburg, Germany 1

Introduction: During allogeneic stem cell transplantation (allo-SCT), infections significantly contribute to morbidity and mortality. We performed a prospective analysis of allo-SCT in 2015 in order to assess epidemiology and outcome of infections during the neutropenic phase. Methods: Data of pts during allo-SCT were recorded in a prospective fashion using a predefined questionnaire. Results: In 2015, 163pts (61f, 102m) were transplanted during 166 procedures. Median age was 59(18-79)y. Median duration of neutropenia was 14.5(4-43)d. Fever (in neutropenia) occurred in 138(117)pts (83.1/70.5%). Fever occurred before an increase in CRP (n = 34; 20.5%), CRP rose before fever (n = 60; 36.1%), CRP and fever co-occurred (n = 44; 26.5%), CRP rose without apparent fever (n = 17; 10.2%) or there was neither an increase of CRP nor fever (n = 11; 6.6%). Severe sepsis developed in 95 (57.2%), septic shock in 26pts (15.7%). BC were drawn in 142pts (85.5%) on a median of 4(1-38)d. At least 1 positive BC was obtained in 34pts (23.9%), 1/2/3/4 BC were positive in 23/7/2/2. In 7/34pts (21%) the same species grew in more than 1 BC (3xStaph.epi.,2xC.paraps.,Crypt.neof.,Ps. aerug.). 1/2/3/4/5/6/9 CVC were inserted in 121/27/10/4/3/1/1pts. The mean duration for the 1st/2nd CVC was 25.6 ± 8.5/13.3 ± 6.2d. At least 1 tCT was done in 94pts, exactly 1/2/3/4/5/6/7 tCT in 34/35/15/5/1/1/3pts. No tCT was necessary for 72pts (43.4%). Lung infiltrates were seen in 64/94pts (68.1%), 51/64 (79.7%) received 1-5 BL. There, Aspergillus antigen or DNA was positive for 14pts. Once, Rhizopus spp. DNA was detected. Fever (n = 73; 44%), fever and increase in CRP (n = 15;9%), increase in CRP (n = 43; 25.9%), or other (n = 35; 21.1%) were reasons for antibiotic therapy. No empiric antibiotic therapy was necessary for 13pts (7.8%). 1st line therapy (CEF,V/D/L) was prescribed in 129pts (77.7%). Escalation to 2nd line therapy (MERO,V/D/L) occurred for 75/129 (58.1%), to 3rd line therapy for 42/75pts (56%). 1st line therapy sufficed for 36pts (21.7%). 2pts died during neutropenia. 145 stays (87.3%) resulted in discharge. 18/21 deaths were due to infectious causes. Conclusion: Immediate broad spectrum antimicrobial therapy, CVC exchange and diagnostic efforts result in infection-related mortality below historical comparatives even in a population of heavy pretreatment and fairly advanced age. Disclosure: Markus Samek: No conflict of interest disclosed. Maximilian Christopeit: Advisory Role: Basilea, MSD; Financing of Scientific Research: Basilea, MSD V46

Community acquired respiratory viruses in healthy controls and allo-SCT recipients – a prospective comparative study Rachow T.1,2, Konowski P.1,2, Kalkreuth J.1, Kurze S.1, Hammersen J.1, Klink A.1, Hilgendorf I.1, Hochhaus A.1, von Lilienfeld-Toal M.1,2 Universitätsklinikum Jena, Klinik für Innere Medizin II, Abt. Hämatologie und Internistische Onkologie, Jena, Germany, 2Leibniz-Institut für NaturstoffForschung und Infektionsbiologie e.V., Hans-Knöll-Institut, Jena, Germany 1

Introduction: Community acquired respiratory viruses (CRV) are an important cause of morbidity and mortality in immunocompromised pa-

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tients, especially after allogeneic hematopoietic stem cell transplantation (allo-SCT). There are just few prospective trials focussing on epidemiological and clinical aspects of CRV in hematologic patients. Methods: Samples of throat gargles were obtained from all participants at 2 or more different time points during the influenza season of each winter regardless of symptoms (for healthy controls 2013/14 and 2014/15 and for patients after allo-SCT 2014/15 and 2015/16). Incidence of CRV and clinical presentation were assessed for both groups. A molecular multiplex PCR assay for 21 respiratory pathogens (FTD® Respiratory Pathogens 21 kit, Fast-track diagnostics Ltd. Malta) was performed for detection of CRV. Results: In the patient cohort 301 throat gargles of 142 patients and in the control group 464 throat gargles of 232 subjects could be analysed. More throat gargles were tested positive for viruses patients when compared to controls (n = 46/301 [15.2%] vs. n = 22/464 [4.7%], p < 0.001). The three most common viruses in the patient cohort were respiratory syncitical virus (RSV, n = 11), adenovirus (n = 8) and coronaviridae (n = 8) whereas in the control group coronaviridae (n = 8), rhinovirus (n = 5) and influenza A (n = 4) were the most common pathogens. Most persons with presence of a virus exhibited symptoms of respiratory tract infection. However, some participants were asymptomatic despite presence of a virus and the rates of asymptomatic virus carriers significantly differed between groups (14/142 patients [9.8%] in patients and 2/232 [0.8%] subjects in the control group, p < 0.001). Conclusions: Patients who have undergone allo-SCT are at higher risk for carrying respiratory viruses. The high rate of asymptomatic carriers might be explained by a prolonged clearance of CRV from the respiratory tract in the condition of immunosuppression. The clinician should keep in mind that even asymptomatic patients after allo-SCT might be a potential source of virus infection for other immunocompromised patients. Disclosure: Tobias Rachow: Financing of Scientific Research: Pfizer; Other Financial Relationships: Reise- und Fortbildungskosten: GILEAD, Baxalta, Medac Marie von Lilienfeld-Toal: Advisory Role: MSD, Celgene, Honorare: MSD, Celgene, Janssen Cilag, Gilead; Expert Testimony: MSD, Pfizer, Deutsche José Carreras Leukämie-Stiftung e.V.; Other Financial Relationships: Reisekosten: Celgene, Janssen Cilag, Gilead, Astellas

Disclosure: Daniel Teschner: Advisory Role: Pfizer Deutschland GmbH, MSD Sharp & Dohme GmbH; Financing of Scientific Research: Gilead Sciences GmbH, MSD Sharp & Dohme GmbH; Other Financial Relationships: Astellas Pharma GmbH , Gilead Sciences GmbH, Jazz Pharmaceuticals Germany, MSD Sharp & Dohme GmbH (Reisekostenerstattungen) Markus Radsak: Advisory Role: Novartis Pharma GmbH; Financing of Scientific Research: Celgene GmbH, Novartis Pharma GmbH; Expert Testimony: Celgene GmbH; Other Financial Relationships: Astellas Pharma GmbH (Reisekostenerstattungen)

Freier Vortrag Rehabilitation V48

Rehabilitation means no risk for patients with hematooncological diseases also and especially after allogeneic blood stem cell transplantation Kiefer-Trendelenburg T.1

V47

The impact of a LysM-specific NFATc1 knockout on neutrophil antifungal defense and myelopoiesis in NFATc1LysM mice infected with Aspergillus fumigatus Teschner D.1, Michel C.1, Prüfer S.2, Theobald M.1, Schild H.2, Radsak M.1 University Medical Center of the Johannes Gutenberg-University, Department of Hematology, Medical Oncology, & Pneumology, Mainz, Germany, 2University Medical Center of the Johannes Gutenberg-University, Institute of Immunology, Mainz, Germany 1

Introduction: Immunosuppressive medication e.g. by calcineurin inhibitors substantially contribute to the risk for opportunistic fungal infections in patients after allogeneic transplantation (HSCT). The nuclear factor of activated T cells (NFAT) is an important transcription factor downstream of calcineurin especially in T cells. Additionally, NFAT also seems to play a substantial role in innate antifungal immune responses by polymorphonuclear neutrophils (PMN), as well as in regulation of myelopoiesis. Methods: LysM-specific NFATc1 knockout (NFATc1LysM) mice were generated by crossing LysM-Cre mice with NFATc1flox/flox mice to obtain NFATc1 deficiency solely in myelomonocytic cells. NFATc1LysM mice were firstly infected with Aspergillus fumigatus (A. f.) conidia intratracheally. PMN recruitment to the lungs, several chemokines/cytokines and pulmonary fungal clearance were examined by analyzing bronchoalveolar lavages (BAL) and peripheral blood (PB) using flow cytometry and cytometric bead array and murine lungs by fungal culture assays and histopathologic examination. In addition, survival was studied with neutropenic animals serving as controls. In addition, we investigated myelopoiesis under steady state conditions by quantifying bone marrow derived myeloid progenitor cells from NFATc1LysM mice using flow cytometry compared to PMN in PB.

Results: NFATc1LysM mice infected with A. f. showed unimpaired survival. However, there were no detectable differences in PMN recruitment or fungal clearance, whereas pulmonary inflammation, chemokines/cytokines production and PMN counts in PB seemed to be more pronounced in knockout mice (0.9 inflammation points/lung ± 0.12 (NFATc1LysM) vs. 0.6 ± 0.08 (control), p = 0.04; 1.5 x 103 PMN/µl ± 0.2 vs. 0.9 ± 0.1, p = 0.04). Distribution of bone marrow derived myeloid progenitor cells was clearly impaired in NFATc1LysM mice especially in megakaryocyte-erythroid progenitor cells (1.2 x 105 cells ± 0.2 vs. 2.7 ± 0.6, p = 0.02) whereas PMN blood counts in PB were unchanged. Conclusion: NFATc1 downregulation in PMN of A. f. infected NFATc1LysM mice leads to enhanced pulmonary inflammation and elevated PMN blood counts compared to controls. Additionally, NFATc1LysM mice display constrained myelopoiesis under steady state conditions without affecting peripheral PMN blood counts compared to wild type controls. Further studies are needed to clarify underlying mechanisms and clinical relevance in HSCT of our findings.

Oncol Res Treat 2016;39(suppl 3):1–104

Klinik am See, Onkologie, Rüdersdorf, Germany

Introduction: Patients with hemato-oncological diseases show a weakened immunsystem, which is especially pronounced after high-dose chemotherapy followed by allogeneic blood stem cell transplantation (HDC/BSCT). To avoid infections patients are recommended to limit contact with other people. In this context, the question arises to what extent the stay in a rehabilitation clinic means a hazard, or whether it may be recommended. Methods: We report a total of 94 rehabilitation stays (RSs) of 91 patients with hemato-oncological diseases, who have been treated under special hygienic conditions during the period from 01-July-2014 to 31-Dec-2015. As the criterion for serious complication early termination of RS has been selected and risk factors were analyzed. Results: Eight of the 94 RSs (8.5%) had to be terminated prematurely. The RSs of patients with prior HDC/alloBSZT had to be discontinued in 21.4% (6 of 28). In comparison the drop-out rate of RSs of patients with so-called solid tumours is 2.4% (29 of 1220). The reasons for stopping RSs were cytomegalovirus reactivation in three patients and in three other cases the occurrence of fever immediately after the start of RS (within the first 48 hours). One can assume, that the patients have been infected before starting RS. In one case RS could not be started due to a low number of leukocytes. The RS of the last patient has to be stopped after one week, since the reduced general condition, vomiting and malnutrition could not be improved. Analyzing possible risk factors for premature termination the platelet count and the period of time between the last treatment and the start of the RS show significant correlation. Age, sex, body mass index, kidney function, CRP and leukocyte count show no significant correlation.

Abstracts

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Conclusions: Since all reasons for premature rehabilitation stop were not caused by the RS itself, we conclude that RSs do not mean a risk for patients with hemato-oncological diseases, assuming an individually tailored hygiene program. On the contrary during a RS life-threatening complications can be detected early and appropriate treatment can be initiated. Disclosure: No conflict of interest disclosed. V49

Oscillating pole therapy – The best option to treat urinary incontinence after radical prostatectomy? - Follow-up-data Heydenreich M.1, Zermann D.-H.1 Vogtland-Klinik Bad Elster, Urologie/ Uroonkologie, Bad Elster, Germany

Introduction: There is evidence that specialized continence training in combination with an oscillation pole therapy has a major effect on the early recovery of continence after prostatectomy. Aim of this investigation was the question, if the results of a prospective randomized controlled study can be confirmed in rehabilitation daily routine. Methods: 336 patients (Ø 64.1 years) with urinary incontinence after prostate cancer surgery were evaluated. All patients passed a standard treatment program (continence training, endurance training, moderate strength training). Additionally the intervention and follow-up group participated in a guided training with an oscillating pole (daily, 30 minutes). Urinary incontinence was evaluated using 1-h (ICS) and 24-hour pad-test. Results: The data of 336 participants could be evaluated. We report the follow-up-data of 168 patients within the framework of normal rehabilitation in comparison to the results of the prospective randomized study published last year.

sical physiotherapeutic treatment (PT) in comparison to a multimedia sensor-based practice on psychological aspects and quality of life (QOL). Methods: Patients undergoing SCT were randomized into the control group (n = 23) receiving PT or the experimental group exercising on the Nintendo-Wii® (n = 19). Patients of both groups completed the FACTBMT, HADS-D and Distress Thermometer at the date of hospital admission (T1) and on day 14 (T2), 28 (T3) and 100 (T4) after SCT. Results: The level of distress was comparable between both groups. However, at T2 distress increased above the cut-off level of 5 in both groups (Wii-group p = 0.006, PT-group p = 0.276). This was accompanied by an increase of anxiety (p = 0.705) and depression (p = 0.006) in the PTgroup, while both parameters decreased in the Wii-group (p = 0.087 and p = 0.220), respectively. Of note, the level of anxiety in the Wii-group stayed below the PT-group at all times. Concerning QOL, social and emotional well-being (SWB/EWB) scored highest in both groups. In Wii-group EWB increased significantly between T1-T4 (p = 0.015) and ranked above PT-group at all times. Physical well-being (PWB) showed the strongest fluctuation of all domains. It declined significantly between T1-T2 in both groups (PT p = 0.015, Wii p = 0.019), followed by a significant increase between T2-T4 (both groups p = 0,001). However, only in Wii-group results of PWB at T4 ranked significantly above T1 (p = 0.028). Lowest scores were proved for functional well-being (FWB) in both groups at all times. The score of bone marrow transplant scale (BMTS), the second lowest score in both groups, was always higher in Wii-group. Conclusion: The results indicate less psychological distress and higher QOL after SCT when exercising with Nintendo-Wii®, which therefore should be considered as an alternative of physiotherapy. Disclosure: No conflict of interest disclosed.

Tab. 1 to V49. Follow-up and study data

Parameter 1-h-PADTest 24-h-PADTest

Control Group n = 84 Begin End

Study Group n = 84 Begin End

Follow-up Data n = 168 Begin End

Significance

22,7 g

17,9 g

22,8 g

28,1 g

p < 0,001

253,3 g

179,3 g 245,3 g

8,6 g

14,3 g

127,7 g 282,5 g

Conclusion: Results of urinary continence therapy after prostate cancer surgery were significantly improved up by complex approach combining standard continence therapy and the new training using an oscillating pole. After integration of the new therapeutic option in rehabilitation daily routine the primary study results were confirmed. Therefore this new treatment option should be offered to all patients suffering urinary incontinence. Disclosure: No conflict of interest disclosed.

152,6 g p < 0,001 V51

Reducing the symptoms of chemotherapy-induced peripheral neuropathy with specific exercise interventions Streckmann F.1,2, Bloch W.2, Lehmann H.3, Faude O.1, Baumann F.T.2 Universität Basel, Department für Sport, Bewegung und Gesundheit, Basel, Switzerland, 2Deutsche Sporthochschule Köln, Institut für Kreislaufforschung und Sportmedizin, Köln, Germany, 3Universitätsklinikum Köln, Neurologie, Köln, Germany 1

Introduction: Hematopoietic stem cell transplantation (SCT) causes severe physical and psychological side effects. However, sports therapy may have positive impact on sentiment. Here we evaluate the influence of clas-

Chemotherapy-induced peripheral neuropathy (CIPN) is a highly prevalent and clinically relevant side-effect of chemotherapy. Depending on the neurotoxic agent, 60-90% of patients are affected. The motor and sensory symptoms not only severely diminish patients’ quality of life, but represent a decisive limiting factor for medical therapy, consequently affecting the clinical outcome. To date approved and effective treatment options are lacking. Promising results have now been achieved with specific exercise interventions. In a first randomized, controlled trial (RCT) with lymphoma patients, positive effects on motor as well as sensory symptoms of CIPN could be achieved with an exercise intervention, concomitant to therapy, comprising endurance-, strength- and sensorimotor training (SMT). Patients, who had exercised, showed improved peripheral deep sensitivity, balance control and level of activity, which enhanced their quality of life. Furthermore, 87% of the patients in the intervention group were able to reduce the symptoms of CIPN, while they persisted in the control group (0%). A tendency towards a potentially preventive effect could also be detect-

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

V50

Prospective, randomized evaluation of the influence of sports therapy on quality of life after hematopoietic stem cell transplantation Stüwe S.1, Schumacher H.1, Kropp P.2, Diedrich D.3, Greger N.1, Freitag S.1, Junghanss C.1, Hilgendorf I.1,4 University Medicine Rostock / Department of Internal Medicine III, Rostock, Germany, 2University Medicine Rostock / Department of Medical Psychology and Medical Sociology, Rostock, Germany, 3University Medicine Rostock / Institute for Biostatistics and Informatics in Medicine and Ageing Research, Rostock, Germany, 4University Hospital Jena / Department of Internal Medicine II, Jena, Germany 1

CONTENTS AUTHOR INDEX

ed. Lacking further references in oncology, we conducted a systematic review, investigating beneficial exercise interventions for patients with peripheral neuropathies independent of the pathophysiology. It revealed that for toxically-induced peripheral neuropathies such as CIPN, exercise interventions involving balance components were essential, while studies conducting endurance- or strength training alone showed no effect on the relevant symptoms. We therefore conducted a further RCT, isolating SMT and comparing it to whole body vibration training (WBV). The two intervention groups were compared to an oncological control group as well as a healthy, age- and gender-matched control group. First analysis showed that both SMT and WBV not only proved feasible for patients with CIPN, but were able to improve reflex activity, peripheral deep sensitivity, balance control and quality of life. We are currently also investigating the preventive effects of SMT and WBV on the incidence of Oxaliplatin- as well as vinca-alkaloid-induced peripheral neuropathy. We therefore propose that SMT as well as WBV are promising exercise interventions to reduce and possibly even prevent symptoms of CIPN. Consequently, specific exercise therapy should be taken more seriously as a supportive therapy for oncological patients. Disclosure: No conflict of interest disclosed. V52

Return to work following a work-related oncological rehabilitation: The pilot study Perspective Job Kähnert H.1, Exner A.-K.2, Muckel E.1, Leibbrand B.3 Institut für Rehabilitationsforschung, Norderney, Bad Salzuflen, Germany, Universität Bielefeld, Fakultät für Gesundheitswissenschaften, AG Epidemiologie & International Public Health, Bielefeld, Germany, 3Salzetalklinik, Bad Salzuflen, Germany 1 2

Introduction: Returning to work is a crucial factor for cancer patients. Accordingly, work-related orientation in medical rehabilitation (MBOR) gains an increasing importance. However, only little is known about contents and processes of MBOR measures in oncological rehabilitation. The aim of the study was to develop and evaluate a multimodal MBOR module called Perspective Job for cancer patients. Furthermore the supporting needs for occupational integration were examined from the patient’s point of view. Methods: Perspective Job was developed by a rehabilitation team. The pilot-study had a sequential control group design with an intervention group (IG: n = 120) and a control group (CG: n = 86). Participants were oncological patients with substantial work-related problems. Patients were asked how helpful they estimated the work-related therapies and defined work-related outcomes (self-assessed working capacity (SWC), functional capability in occupational (FCO), return to work (RTW)) over 3 months after discharge. Furthermore telephone based interviews with participants were conducted 6 and 18 weeks after discharge. Chi-square tests and analysis of covariance were applied to examine differences between CG and IG. Results: Perspective Job consists of occupational therapies and job trainings. The IG emphasized that most therapies were classified as work-related (p < 0.001) than the CG. Three months after discharge, the IG felt better prepared for returning to work (p < 0.05), were considerably less limited in their FCO (p < 0.01) and estimated their SWC more often as re-established (p < 0.05) than the CG. 65% of the IG and 57% of the CG returned to work 3 months after discharge (p = 0.551). The telephone interviews illustrate: A stepwise occupational reintegration is described as an essential instrument for a successful RTW. Moreover, the RTW can be promoted by good health status, work-related rehabilitation with planned occupational aftercare, support by doctors and good working climate. Conclusions: The pilot study provides evidence that Perspective Job is a useful intervention to enhance work-related outcomes for cancer patients with substantial work-related problems. Facilitators for RTW were good functional abilities, health status, supports by doctors and positive working atmosphere. Moreover, a work-related rehabilitation and occupational

Oncol Res Treat 2016;39(suppl 3):1–104

aftercare activities facilitate return to work. Further studies should be carried out to verify these results. Disclosure: Heike Kähnert: No conflict of interest disclosed. Birgit Leibbrand: Employment or Leadership Position: Vorstandsmitglied im Verein zur Förderung der Rehabilitationsforschung e. V. Norderney V53

Sustainability of a urooncological rehabilitation program for prostate cancer patients Heydenreich M.1, Zermann D.-H.1 Vogtland-Klinik Bad Elster, Urologie/ Uroonkologie, Bad Elster, Germany

Introduction: Specialized rehabilitation programs allow an early social and occupational reintegration of patients after prostate cancer surgery. Aim of this prospective study was an evaluation of the sustainability and efficacy of a complex training program two years after rehabilitation. Methods: 150 former patients after prostate surgery were contacted and asked to fill out a questionaire. All former patients had a standardized functionally oriented, specialized 3-week-rehabilitation program completed two years ago. Results: 47 patients (Ø 67.2 years) completed the questionnaire. The following results were obtained: 1. 93,6% of patients had a good to excellent state of health 2. 40,4% of patients were without stress incontinence 3. 53,2% of patients performed continence training 4. number of used pads (Ø 1,15 pads (SD = 1,37)) for security reasons or persistent urinary incontinence 5. the most common daily life activities were: walking (31,4%), cycling (19,8%), gymnastic (14%) and swimming (9,3%) 6. 97,9% of patients evaluated the rehabilitation program with good to excellent 7. 100% of patients evaluated the medical care with good to excellent 8. 100% of patients evaluated the workshops with good to excellent 9. 91,5% of patients would visit Vogtland-Klinik Bad Elster for a second time, if necessary Conclusion: Specialized rehabilitation has a strong sustainability! This applies for all aspects of general health, healthy behavior and also functional deficits. Two years after rehabilitation patients still benefit from rehabilitation after prostatectomy. Therefore rehabilitation should be offered to all prostate cancer patients. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Chronische lymphatische Leukämie – Biologie V56

Genetic loss of NFAT2 leads to CLL transformation Fuchs A.R.1, Märklin M.1, Heitmann J.S.1, Truckenmüller F.M.1, Saur S.J.1, Ganser M.1, Bugl S.1, Kopp H.-G.1, Kanz L.1, Rao A.2, Wirths S.1, Müller M.R.1 Universitätsklinikum Tübingen, Medizinische Klinik und Poliklinik, Abteilung für Onkologie, Hämatologie, Rheumatologie, Klinische Immunologie und Pulmonologie, Tübingen, Germany, 2La Jolla Institute of Allergy and Immunology, La Jolla, United States 1

NFAT2 is a transcription factor which regulates developmental and activation programs in diverse cell types. CLL constitutes a heterogeneous disease with some patients exhibiting an indolent course for many years and others progressing rapidly and requiring early treatment. A defined subgroup of patients shows enhanced responsiveness to stimulation of the B cell receptor (BCR) complex and more aggressive disease. In contrast, another subset of CLL patients with more indolent course is characterized by an anergic B cell phenotype referring to B cell unresponsiveness to IgM ligation. Here, we analyzed the role of NFAT2 in CLL with respect to the

Abstracts

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maintenance of the anergic phenotype and the development of Richter´s syndrome. For this purpose, we generated conditional NFAT2fl/fl CD19-Cre knock out mice on the Eµ-TCL1 transgenic background. These mice exhibit NFAT2 deletion limited to the B cell lineage and develop a human-like CLL. To extend our observations to the human disease, we further analyzed primary blood samples from patients with CLL (n = 30) and Richter´s syndrome (n = 5). Mice with genetic loss of NFAT2 in their B cell compartment showed a significantly more aggressive disease course and a dramatically reduced life expectancy. Spleen and lymph nodes exhibited a transformed disease phenotype in the NFAT2 ko cohort with diffuse distortion of their architecture by large B cells with blastic chromatin. CLL cells with regular NFAT2 expression were unable to mobilize calcium upon IgM ligation and thus exhibited the typical anergic phenotype. Using affymetrix microarrays we defined a genetic signature of anergy in CLL cells consisting of Lck, Pacsin1 and the E3 ligases Cbl-b and Grail, which were highly expressed in CLL cells with normal NFAT2 expression. Furthermore Lck was constitutively activated in anergic CLL cells while it was essentially inactive in NFAT2 ko cells suggesting an important role for Lck in mediating the anergic phenotype. Primary human CLL samples showed a significant overexpression of NFAT2 and Lck, predominantly in its active conformation. Tissue samples from patients with Richter’s Syndrome on the other hand demonstrated a significantly reduced expression of NFAT2 and complete loss of Lck expression. In summary, our data provide strong evidence that NFAT2 and its target gene Lck are important mediators of anergy induction in CLL and that their loss may lead to disease transformation to aggressive B cell lymphoma (Richter´s syndrome). Disclosure: No conflict of interest disclosed. V57

Loss of NFAT2 leads to an acceleration of clonal evolution in CLL Müller D.J.1, Märklin M.1, Kanz L.1, Wirths S.1, Müller M.R.1 Universitätsklinikum Tübingen, Medizinische Klinik und Poliklinik, Abteilung für Onkologie, Hämatologie, Rheumatologie, Klinische Immunologie und Pulmonologie, Tübingen, Germany 1

Chronic lymphocytic leukemia (CLL) may be defined as a clonal expansion of mature B cells with stereotypic B cell receptors (BCR). Somatic hypermutation of the BCR heavy chains (IGVH) defines subgroups with different prognosis. In up to 10% of cases Richter’s transformation to a high grade lymphoma with a mostly dismal prognosis is observed. The TCL1 transgenic mouse is a well accepted model of human CLL. Upon B cell specific knock out of NFAT2, TCL1 mice develop a disease resembling Richter’s syndrome with a significantly more aggressive disease phenotype. While TCL1 B cells exhibit tonic anergic BCR signaling characteristic of human CLL, loss of NFAT2 leads to readily activated BCRs indicating different BCR usage with altered down-stream signaling. Here, we analyzed BCR usage in C57BL/6 wildtype, TCL1, and TCL1NFAT2-/- mice. Splenocyte cDNA of the respective animals at an age of 7 months was amplified by multiplex PCR covering known heavy chain VDJ alleles. The PCR products were subsequently cloned into standard bacterial plasmids and subjected to conventional DNA sequencing. In addition, PCR products were analyzed by next-generation-sequencing (NGS). For data analysis, the IMGT/HightV-QUEST online tool was applied. A very diverse and polyclonal BCR usage was found in wild type mice with more than 4000 different clones identified. Although TCL1 mice at 7 months of age exhibit all features of CLL, their BCR usage as analyzed by unambiguously identified VDJ recombination was still polyclonal. Only with respect to VH usage, TCL1 mice were found to use a more limited set of V alleles compared to wild type mice. Loss of NFAT2 by conditional knockout on the contrary, leads to an oligoclonal usage of VDJ recombination, to a further limitation of V alleles, and also to the usage of identical VDJ recombination in different mice with high frequency – indicating

Abstracts

BCR selection either by antigen or self-signaling in these mice. Further analyses by sequencing of hypervariable regions showed preferential usage of mutated BCRs in TCL1 mice and of unmutated BCRs in NFAT2 ko mice reflecting benign and aggressive disease in humans. In summary, the loss of NFAT2 signaling in CLL precipitates the selection of unmutated BCRs as well as the preferential usage of certain VDJ recombinations which results in the accelerated development of true oligoclonal disease. Disclosure: No conflict of interest disclosed. V58

Knock-down of the NFAT2 long and intermediate isoforms leads to CLL acceleration Heitmann J.S.1, Märklin M.1, Truckenmüller F.M.1, Fuchs A.R.1, Kopp H.-G.1, Kanz L.1, Rao A.2, Wirths S.1, Müller M.R.1 Universitätsklinikum Tübingen, Medizinische Klinik und Poliklinik, Abteilung für Onkologie, Hämatologie, Rheumatologie, Klinische Immunologie und Pulmonologie, Tübingen, Germany, 2La Jolla Institute of Allergy and Immunology, La Jolla, United States 1

NFAT2 is a highly phosphorylated transcription factor which regulates developmental and activation programs in diverse cell types. We and others have previously described a significant overexpression of NFAT2 in CLL cells as compared to physiological B cells. Three major isoforms of NFAT2 with different regulatory properties have been described (700 aa short isoform, 800 aa intermediate isoform, 900 aa long isoform). Here, we analyzed the role of different NFAT2 transcripts in CLL with respect to disease phenotype and cell proliferation. We investigated primary samples from CLL patients (n = 30) for their expression profile of different NFAT2 isoforms using RT-PCR. Applying an shRNA approach, we generated stable knock-down cells of the CLL cell line MEC-1 for the long and intermediate isoforms and for the entire NFAT2 gene resulting in the complete ablation of all isoforms. The proliferation properties of the different MEC-1 cell line were subsequently assessed in xenotransplant experiments into NSG mice. While physiological B cells express comparable levels of the short and intermediate/long isoforms, we could detect a fivefold overexpression of the intermediate/long isoforms in primary CLL samples. To further analyze the differential regulation of the different NFAT2 transcripts on tumor cell proliferation and cell cycle regulation, we injected NSG mice with MEC-1 cells with intact NFAT2 (n = 6), MEC-1 cells with a knock-down of the intermediate and long isoforms (n = 6) and MEC-1 cells with a complete NFAT2 knock-down (n = 6). MEC-1 cells with selective ablation of the intermediate and long NFAT2 isoforms grew significantly faster in NSG mice than MEC-1 cells with intact NFAT2 expression or MEC-1 cells with a complete NFAT2 knock-down. MEC-1 cells selectively lacking the intermediate and long isoforms led to accelerated tumor proliferation upon subcutaneous injection. Cell cycle analysis as assessed by flow cytometry showed a significantly increased number of cells in the G1/S-Phase for the group without expression of the short isoform, while the group with complete NFAT knock-down exhibited a compromised growth pattern as compared to wild-type MEC-1 cells. In summary, our data demonstrate that genetic loss of the intermediate and long isoforms of NFAT2 leads to CLL acceleration Disclosure: No conflict of interest disclosed.

Oncol Res Treat 2016;39(suppl 3):1–104

CONTENTS AUTHOR INDEX

V59

AKT activation in Chronic Lymphocytic Leukemia cells promotes transformation towards aggressive Richter´s syndrome lymphoma Nickel N.1, Al-Maarri M.2, Pal M.2, Roth A.1, Schäfer S.3, Büttner R.3, Hallek M.1, Wunderlich T.2, Pallasch C.P.4 Klinik I für Innere Medizin, Universität Köln, CIO, Köln, Germany, 2MPI für Stoffwechselforschung, Köln, Germany, 3Institut für Pathologie, Köln, Germany, 4 Klinik I für Innere Medizin, Universität Köln, CIO, CECAD, Köln, Germany 1

Introduction: Richter´s syndrome (RS) is an aggressive transformation of Chronic Lymphocytic Leukemia (CLL) refractory to current therapies with dismal prognosis. RS arises from CLL cells independent of common DLBCL-mutations. Frequently mutations in p53, CDKN2 or cMyc genes are involved, but a significant proportion displays no specifically acquired driver mutation. Here we aim to elucidate the role of AKT in transformation towards Richter´s syndrome in human patients and functionally address constitutively AKT-activation in vivo. Ultimately, we aim to develop mouse models that can be used to develop novel treatment options towards Richter´s syndrome. Methods: We have developed a mouse model allowing for Cre-activatable expression of a myristoylated AKT constitutive active allele from the Rosa26 locus (AKT-C). We crossed that mice with Cd19Cre- and Cg1Cre-mediated B cell-specific AKT-C expression in the Eµ:Tcl1 CLL mouse model. Results: In biopsies revealed from patients with RS that one third of cases showed enhanced AKT activation. Strikingly, Richter´s transformation was recapitulated when AKT was genetically over-activated in Eµ-Tcl-1 mice, where AKT-C expressing cells developed a high-grade lymphoma phenotype leading to significantly decreased survival. AKT-C expression in TCL1 CLL cells furthermore induced morphology changes displaying features of aggressive lymphoma such as large transformed B-cell phenotype and frequent mitotic figures, enhanced proliferation indicated by KI67. Thus, Eµ:Tcl1 transformed CLL cells act in concert with constitutively active AKT to develop RS. Noteworthy, Cd19Cre;AKT-C double transgenic mice fail to develop leukemia and lymphoma, indicating that CLL development in Eµ:Tcl1 mice is a precondition to transformation. As for the downstream mechanisms of AKT-mediated transformation we identified GSK-3b inhibition and subsequent cMyc and Mcl-1 stabilization. This might confer contribute to resistance of AKT-C transfomed lymphoma cells against genotoxic and targeted drugs. Here we see resistance towards conventional chemotherapeutics as against BTK/PI3K/BCL2-inhibiting compounds in this novel model of Richter´s syndrome. Conclusions: Collectively, we have generated the first murine RS model providing novel mechanistic insights into the molecular understanding of Richter´s transformation that is amenable to model therapeutic strategies and to address the efficacy of synergistic treatment combinations. Disclosure: Nadine Nickel: No conflict of interest disclosed. Christian Pallasch: Advisory Role: Gilead Sciences V60

The microenvironment alters CLL cell response to NF-kB inhibition Foerster K.1, Simon-Gabriel C.-P.1, Bleckmann D.1, Benkisser-Petersen M.1, Thornton N.1, Claus R.1, Duyster J.1, Zirlik K.1 University Medical Center Freiburg, Department of Hematology, Oncology, and Stem cell transplantation, Freiburg, Germany 1

Introduction: NF-kB transcription factor is constitutively active in CLL and has been suggested as promising therapeutic target. NF-kB inhibition induces apoptosis in CLL cells in vitro. However, whether this effect pertains in vivo remains unclear. Since the microenvironment is crucial for CLL cell viability circumventing apoptosis, we tested whether NF-kB inhibition modulates CLL viability in the presence of microenvironment.

Oncol Res Treat 2016;39(suppl 3):1–104

Methods: The specific NF-κB inhibitor Dehydroxymethylepoxyquinomicin (DHMEQ) was used alone or combined with fludarabine, BAFF, APRIL, CXCL12, or CD40 ligand on primary CLL cells in mono- or in co-culture with stromal cells (BMSC). Apoptosis was measured using AnnexinV/PI stainings. Protein expression was analyzed by western blot. NF-kB DNA-binding activity was measured by ELISA and p65 knockdown was performed by siRNA transfection. Results: NF-kB inhibition using DHMEQ led to apoptosis in monocultured CLL cells (viability 74% vs. 24%, n = 17, p < 0.0001) but surprisingly had no effect on cell viability of cells co-cultured with BMSC (viability 96% vs. 95%, p = 0.9995). In monoculture, apoptosis induction was accompanied by downregulation of the NF-kB target protein TRAF-1, upregulation of the proapoptotic protein Bax, and increased PARP cleavage. Conversely, in co-culture, downregulation of TRAF1 and several NF-kB subunits were observed without concomitant Bax upregulation or PARP cleavage. NF-kB DNA-binding activity of all NF-kB subunits was equally suppressed by DHMEQ treatment in mono- and co-cultured cells. Knock down of the NF-kB subunit p65 solely induced apoptosis in monocultured CLL cells. Adding soluble BAFF to monocultured CLL cells, attenuated DHMEQ efficiency (viability 1% vs. 16%, n = 9). Finally, the combined use of DHMEQ with fludarabine in co-cultured CLL cells led to a higher rate of apoptosis than DHMEQ (viability 57% vs. 37%, p = 0.0202) or fludarabine alone (viability 50% vs. 37%, p = 0.1828). Conclusion: NF-kB inhibition in primary CLL cells shows great discrepancy between in vitro and in vivo scenarios. While DHMEQ treatment leads to apoptosis in mono-cultured cells, CLL cell viability is not affected in the presence of microenvironment, suggesting that the NF-kB pathway can be bypassed in vivo. Soluble ligands appear to be involved in mediating this protective effect. However, the combination of NF-kB inhibition with standard chemotherapy might represent a fruitful approach and warrants further clinical assessment. Disclosure: No conflict of interest disclosed. V61

Pathway inhibitors for bridging to allogeneic hematopoietic stem cell transplantation (HSCT) for chronic lymphocytic leukemia (CLL) Hahn M.1, Dietrich S.1, Hain S.-A.2, Hegenbart U.1, Rieger M.3, Scheuer B.4, Ho A.D.1, Dreger P.1 Universität Heidelberg, Medizinische Klinik V, Heidelberg, Germany, 2St. Marien-Krankenhaus, Medizinische Klinik III, Siegen, Germany, 3Onkologische Schwerpunktpraxis, Darmstadt, Germany, 4Onkologische Praxis, Pirmasens, Germany 1

Pathway inhibitors (PI), such as ibrutinib and idelalisib, have dramatically increased the treatment options for high-risk CLL. There is only limited information on the feasibility and efficacy of HSCT if used as consolidating treatment after PI salvage for relapsed/refractory high-risk CLL. Aim: of this single centre retrospective analysis was to investigate the outcome of HSCT after previous PI exposure in patients with CLL. Results: Between Nov 2013 and Feb 2016, 9 patients (m/f= 6/3; median age 53y (33-61)) underwent HSCT for relapsed/refractory high-risk CLL (RR-CLL) after prior exposure to ibrutinib (6), CC-292 (2), idelalisib (2), and/or venetoclax (1) (multiple exposure included). Indications for HSCT were (hierarchical) RR-CLL with poor-risk cytogenetics (6), previous PI failure (1), and Richter transformation (RT) (2). Before start of conditioning, 8 patients were still responding to the most recent PI with incipient progression under ibrutinib in 2 of them. A single patient with RT in CR was off PI. Patients had been on their most recent PI for 9 (1-15) months. Donors were well-matched unrelated (8) or related (1); conditioning comprised fludarabine/ATG with Treosulfan (6) or TBI 8Gy (3). PI was generally stopped on the day before conditioning. However, a single patient developed a fulminant CLL relapse under fludarabine conditioning within 3 days after ibrutinib discontinuation and was put back on ibrutinib until day -1. With 10 (2-22) months of follow-up, low-grade acute and chronic GVHD each occurred in one patient. Whilst unexpected toxicity was not

Abstracts

CONTENTS AUTHOR INDEX

observed, one patient who had been bridged with ibrutinib dynamically progressed at day +72, and another patient had a relapse of her RT on day +214 resulting in rapid death. The 7 remaining patients live progression-free, 5 of them MRD-negative. Conclusion: This preliminary data does not raise safety concerns about the use of PI prior to HSCT. Ibrutinib seems to be effective for bridging patients with high-risk CLL to transplant. Although disease control after HSCT does not appear to be affected by pre-transplant PI exposure, the optimum strategy for transition from PI treatment to transplant remains to be defined. Disclosure: Michael Hahn: No conflict of interest disclosed. Peter Dreger: Advisory Role: Janssen, Gliead; Financing of Scientific Research: Janssen, Gilead

Fortbildung

Indolente Non-Hodgkin-Lymphome V63

WHO-Update 2016: What is relevant? Rosenwald A.1 Institut für Pathologie, Universität Würzburg, Würzburg, Germany

The 2016 revision of the World Health Organization (WHO) classification of lymphoid neoplasms reflects the progress that has been made in our biological, genetic and clinical understanding of some lymphoma entities over the last years impacting on the development of more targeted therapeutic strategies in the future. In the arena of indolent Non-Hodgkin lymphomas, the concept of ‘precursor’ or ‘early’ lesions as well as very indolent forms of the disease is emphasized. Monoclonal B-cell lymphocytosis (MBL) is now recognized as a precursor of chronic lymphocytic leukemia (B-CLL). However, ‘low count’ MBL that very rarely progresses to overt B-CLL is now distinguished from ‘high count’ MBL, since the latter has many phenotypic, molecular and genetic features of Rai stage 0 B-CLL. The concept of ‘tissue-based’ MBL (discrete lymph node involvement by SLL cells) with a low rate of progression is introduced. Precursor lesions of follicular lymphoma (FL) and mantle cell lymphoma (MCL) have been renamed to ‘in situ follicular neoplasia (ISFN)’ and ‘in situ mantle cell neoplasia (ISMCL)’ to reflect the low rate of progression to overt lymphoma. The ‘duodenal type’ of FL is specifically mentioned, since it appears to remain localized for a long period and to be associated with an excellent outcome. The previous term of ‘pediatric FL’ will be changed to ‘pediatric-type FL’ to reflect the finding that these B-cell proliferations that are almost always localized can occasionally occur also in adults. Finally, novel genetic findings that were made possible by modern next generation sequencing technologies are introduced in the 2016 revision. Almost all cases of hairy cell leukemia carry the BRAF V600E mutations, whereas this mutation appears to be absent in hairy cell leukemia variant which in half of the cases carries a MEK1 mutation instead. Lymphoplasmacytic lymphoma (LPL, Waldenström macroglobulinemea) is associated in about 90% of cases with MYD88 (L265P) mutations. In this context, monoclonal gammopathy of undeterminded significance (MGUS) of IgM-type may be more closely related to LPL than to plasma cell myeloma. In summary, the 2016 update of the WHO classification provides updated diagnostic lymphoma categories including their precursor lesions, together with the description of more refined biological and clinical correlates. Disclosure: No conflict of interest disclosed.

Abstracts

V64

Follicular Lymphoma Grade 3 - To CHOP or not to CHOP? Scholz C.W.1 Vivantes Klinikum Am Urban, Hämatologie und Onkologie, Berlin, Germany

Follicular lymphoma (FL) is the most prevalent indolent lymphoma. Depending on the frequency of interspersed centroblasts, FL are divided into grade 1 to 2 (grade 1/2), if less than 16 centroblasts per high power field (HPF) are detected among centrocytes, and grade 3, if more than 15 centroblasts are counted per HPF. Grade 3 is further grouped into grade 3A, if neoplastic follicles are composed of centroblasts and centrocytes, and grade 3B, if only centroblasts are detectable. Noteworthy, CD10 expression and t(14;18)/BCL2 rearrangement, two distinguishing marks of FL, are less frequently found in FL grade 3 as compared to grade 1/2. Remarkably, FL grade 3A frequently contains areas of grade 1/2 in the same specimen, while this rarely is the case for FL grade 3B. This circumstance likely further increases interobserver variability when it comes to diagnosing FL grade 3. Therapy regimen for individuals with FL grade 1/2, which account for approximately 85% of all FL patients, have been investigated in distinct phase II and III trials, challenging the need for anthracyclines in first line treatment. As a consequence, Rituximab Bendamustin has replaced R-CHOP as first line therapy in many centers. In contrast, treatment for patients with FL grade 3 is far less evidence based, as most prospective clinical trials with FL patients either exclude individuals with FL grade 3 or contain only small subsets of patients with FL grade 3A or 3B. FL grade 3B is deemed to be a more aggressive disease and therefore is treated like diffuse large B cell lymphoma with R-CHOP. The same is applicable for cases where a diffuse large B cell lymphoma component is detected concomitantly with FL grade 1/2 or 3. In contrast, the situation is less settled for FL grade 3A, which account for approximately 10% of all FL. Here, evidence resulting from retrospective register analyses and post-hoc evaluations of prospective trials, suggests that patients with FL grade 3A, progress similar to or, in respect to patients with pure FL grade 3A, fare better than patients with FL grade 1/2. While many of the individuals from these analyses received anthracycline-containing regimen like CHOP or R-CHOP as first line treatment, it remains unresolved, whether anthracyclines are truly required for first line therapy of all patients with FL grade 3A, or whether for some of them a regimen like Rituximab Bendamustin could be an alternative similar to the situation in FL grade 1/2. Disclosure: Christian Scholz: Advisory Role: Gilead, Janssen-Cilag, Novartis, Roche, Sepropharm; Financing of Scientific Research: Gilead, Janssen-Cilag, Novartis, Pfizer, Roche, Sepropharm; Expert Testimony: Mundipharma, Novartis V65

Mantle cell lymphoma: has the era of immun-kinomchemotherapy come? Hess G.1 III. Medizinische Klinik, Universitätsmedizin Mainz, Mainz, Germany

Mantle cell lymphoma still has a dismal prognosis, however new treatment approaches have entered the therapeutic arena recently with the potential to ultimately improve prognosis of patients: optimized use of conventional immune-chemotherapy and secondly the introduction of more specific targeted agents. Among the first group the introduction of cytarabinoside in first line treatment combined with dose intense consolidation treatment combined with antibody treatment has resulted in unprecedented long term responses. In addition early results show that maintenance treatment using rituximab may further improve PFS and potentially OS. For patients not suitable for intensive treatment receiving R-CHOP, R-maintenance already can be considered standard of care, whereas optimal regimen incorporating cytarabinoside need yet to be defined for this group. As the susceptibility to chemotherapy approaches is in general limited to very few treatment lines, alternative approaches have been investigated early on, proofing the suitability of drugs like bortezomib, lenalidomide and temsirolimus. All of these drugs were able to induce reasonable re-

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CONTENTS AUTHOR INDEX

sponses in relapsed disease, however responses to single agent treatment used to be short lived. Ibrutinib now has become the best targeted agent available with responses observed in up to 70% of patients and a median remission duration for more than one year. Although superior to other treatments, still this cannot be considered optimal and attempts have been initiated to improve the use of these agents. In general 2 approaches have been used: development of so called chemofree regimen and all-in combinations testing these agents together with chemotherapy and immunotherapy. The first type of combinations aims to achieve optimized response rates and remission duration primarily with treatment until progression und comparison is made against conventional regimen in terms of efficacy and side effects. Trials like the ENRICH (Ibr + R vs. Chemo + R)( will help to understand the value of these approaches. In contrast, approaches combining all available treatment modalities try to achieve deep responses with limited treatment duration. Key trials to investigate these concepts are the TRIANGLE (intensive treatment +/Ibr) trial and to some extent the SHINE (BR+/- Ibr) trial. Taken together, improvements have been made using all available therapeutic options, with the optimal use still needs to be defined.

V67

Chemotherapeutic treatment of cancer of unknown primary Krämer A.1 Klinische Kooperationseinheit Molekulare Hämatologie/Onkologie, Medizinische Klinik V, Universität Heidelberg und Deutsches Krebsforschungszentrum, Heidelberg, Germany 1

CUP: Molecular Diagnostics

Cancer of unknown primary (CUP) is a heterogeneous group of histologically confirmed metastatic cancers for which the anatomical site of origin remains occult despite appropriate examination. Approximately 3-5% of all human cancers manifest as CUP, which is the seventh most frequent cancer diagnosis and the fourth most common cancer-related cause of death in patients of both sexes. CUP can be divided into favorable and unfavorable subsets. Patients with unfavorable CUP account for 70-80% of cases and have a poor prognosis with a median survival of 8-11 months. Two opposing hypotheses exist with regard to the biology of CUP, which is still poorly understood. One hypothesis considers CUP to be a separate type of cancer with unique biologic features that account for the absence of a primary tumor and early metastatic disease. Based on this view, many patients with unfavorable CUP uniformely receive platinum-based standard chemotherapy in combination with a taxane or gemcitabine. Alternatively, CUP represents different, site-specific, unrelated groups of malignancies. In that case, gene expression profiling approaches to identify the tissue-of-origin might allow to extend the management of known cancers to subtypes of CUP in order to advance therapies for this disease. Most recently, mutational profiling has been suggested to alleviate the need for tissue-of-origin identification, making CUP an epitome of personalized medicine, with individualized treatment driven by the mutational status rather than the primary site of each patient.

Weichert W.1

Disclosure: Alwin Krämer: Advisory Role: Roche; Expert Testimony: Merck

Disclosure: Georg Hess: Advisory Role: Pfizer, Celgene, Roche, Janssen; Expert Testimony: Pfizer, Roche, CTI, Celgene

Fortbildung CUP-Syndrom V66

Technische Universität München, Institut für Pathologie, München, Germany

Carcinoma of unknown primary (CUP) is a challenging disease “entity” not only on the clinical side but specifically when it comes to the selection of diagnostic modalities to apply in this setting. Diagnostically, initial conventional histology aided by immunohistology aims at diminishing the number of cases, which have to be assigned as CUP and helps to identify putative primaries. Although this approach is not successful in all cases, it might still delineate a groups of special CUP cases with likely primary sites. However, the majority of CUP cases remain entirely non-classifiable and usually present as adenocarcinomas with an uncharacteristic immune phenotype. For these cases multimarker expression analyses investigating either mRNA or microRNA profiles have been proposed as an adjunct for diagnosis but these approaches have failed to enter clinical routine testing so far. Recently, massive parallel sequencing (MPS) approaches have enabled us to interrogate comprehensive molecular profiles from human tumors and it has been shown that this technology can be implemented into the routine diagnostic setting. For CUP cases MPS approaches are specifically interesting, since they a) might supply additional information with respect to entity classification and b) might detect molecular alterations that are indicative for the response to specific targeted drugs. Consequently, targeted multigene and broader exomic/genomic sequencing approaches have been used to define the molecular landscape of these tumors and aid in CUP therapy selection. The respective research approaches and available data are discussed.

Fortbildung Lebertumoren V69

Rational imaging of hepatic lesion: How to do it Wiggermann P.1 Universitätsklinikum Regensburg, Institut für Röntgendiagnostik, Regensburg, Germany 1

The purpose of this lecture is to address capabilities and limitations of different imaging techniques such as ultrasound (US), contrast enhanced US (CEUS), magnetic resonance imaging (MRI), computed tomography (CT) and positron emission tomography-CT (PET-CT) for detection and differentiation of hepatic lesions. Relevant implications from national and international guidelines for different tumor entities regarding hepatic imaging will be discussed and set into a framework for a rational imaging pathway of hepatic lesions in a clinical setting. Disclosure: Philipp Wiggermann: Expert Testimony: Bayer Vital GmbH - 51368 Leverkusen

Disclosure: No conflict of interest disclosed.

Oncol Res Treat 2016;39(suppl 3):1–104

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CONTENTS AUTHOR INDEX

Wissenschaftliches Symposium

Neue immuntherapeutische Ansätze nach allogener Stammzelltransplantation V75

Gamma-delta T-cells: graft-versus-leucemia effect without graft-versus-host disease? Kunzmann V.1, Smetak M.2, Schaefer-Eckart K.2, Kimmel B.1, Birkmann J.2, Einsele H.1, Wilhelm M.2 Medizinische Klinik und Poliklinik II, Universitätsklinikum Würzburg, Hämatologie/Onkologie, Würzburg, Germany, 2Klinikum Nürnberg, Medizinische Klinik 5, Nürnberg, Germany 1

Introduction: Human gammadelta (γδ) TCR-expressing cells constitute 1-5% of total peripheral T cells and have been shown to exert potent antitumor activity in a MHC-independent manner. Recent analysis of various cancer types revealed the presence of intratumoral γδ T cells as the most significant favourable prognostic immune population. Within this pilot study we want to determine the feasibility and safety of an adoptive transfer and in vivo expansion of human haploidentical γδ T lymphocytes. Methods: Four patients with advanced haematological malignancies (AML, NHL or Multiple Myeloma) who were not eligible for allogeneic transplantation received peripheral blood mononuclear cells from haploidentical family donors. For that, a single unstimulated leukapheresis product was incubated with both the anti-CD4 and anti-CD8 antibodies conjugated to paramagnetic particles. On average, patients received 2.17 × 106/kg (range 0.9-3.48) γδ T cells with < 1% CD4-or CD8-positive T cells remaining in the product. All patients received prior lymphopenia-inducing chemotherapy (fludarabine 20-25 mg/m2 day -6 until day -2 and cyclophosphamide 30-60 mg/kg day -6 and -5) and were treated with 4 mg zoledronate on day 0 and 1.0×106 IU/m2 IL-2 on day +1 until day +6 for the induction of γδ T cellproliferation in vivo. Results: This treatment resulted in a marked in vivo expansion of donor γδ T cells and, to a lower extent, natural killer cells and double-negative αβ T cells (mean 68-fold, eight-fold, and eight-fold, respectively). Proliferation peaked by around day +8 and donor cells persisted up to 28 days. Although refractory to all prior therapies, three out of four patients achieved a complete remission, which lasted for 8 months in a patient with plasma cell leukaemia. None of the patients showed any signs of acute or chronic GVHD. Conclusions: This pilot study shows that adoptive transfer and in vivo expansion of haploidentical γδ T cells is feasible and suggests a potential role of this donor γδ T cell infusion in the treatment of haematological diseases. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium

ALL: Molekulare und zelluläre Grundlagen für zielgerichtete Therapien V77

Clonal heterogeneity and evolution in acute lymphoblastic leukemia Brüggemann M.1 Sektion für Hämatologische Spezialdiagnostik, Klinik für Innere Medizin II, UKSH, Campus Kiel, Kiel, Germany 1

Recent genomic studies have provided insight into tumor heterogeneity of acute lymphoblastic leukemia (ALL) and the evolutionary trajectory from diagnosis to relapse. In fact, continuing clonal expansion itself might be accompanied by genetic and functional diversification of cancer cells. Their subclonal diversity is inter alia held accountable for primary or secondary resistance to systemic therapy. Clonal heterogeneity and subclon-

Abstracts

al hierarchies have been substantially studied particularly in childhood BCR-ABL fusion positive (BCR-ABL+) BCP-ALL: linear and branching patterns of clonal evolution have been documented and diverse individual subclones showed leukemia maintaining or initiating potential in xenograft models. Furthermore, founder mutations like the BCR-ABL fusion may originate at distinct stages of hematopoietic development. All these findings translate into a clinically heterogenous disease also regarding treatment response and prognosis. Molecular analyses of other recurrent genetic alterations like intrageneic deletions of the BTG1, IKZF1 and ERG indicate that also these aberrations can arise independently in multiple subclones. Detailed sequence analyses of intrageneic breakpoints strongly suggest illegitimate RAG1/RAG2-mediated recombination as the responsible mechanism for aberrations. Comparison of the clonal architecture of ALL between diagnosis and relapse may help to characterize the prognostic meaning of subclonal events for treatment resistance as resistant mutated subclones may survive therapy, acquire additional mutations and become the relapse founder clone. In childhood all, NT5C2, CREBBP, WHSC1, TP53, USH2A, NRAS and IKZF1 are enriched at relapse indicating their potential importance for drug resistance. E. g. recurrent NT5C2 mutations potentially allow for resistance to nucleoside analogs. These mutations predominantly emerge early during maintenance therapy in which nucleoside analogs assume a predominant role in treatment. In summary, analysis of the dynamics of the subclonal structure of ALL and may help to get insights into the biological meaning of particular aberrations and to optimize therapy accordingly. Disclosure: No conflict of interest disclosed. V78

Personalized medicine for ALL? The pediatric INFORM project Worst B.C.1,2, van Tilburg C.M.1,2,3, Balasubramanian G.P.1, Fiesel P.2, Pfaff E.1,2, Pajtler K.1, Freitag A.3, Witt R.3, Kulozik A.E.2, von Stackelberg A.4, Meisel R.5, Borkhardt A.5, von Deimling A.1,2, Eggert A.4, Lichter P.1, Capper D.1,2, Witt O.1,2, Pfister S.M.1,2, Jones D.T.W.1 Deutsches Konsortium für Translationale Krebsforschung (DKTK) und Deutsches Krebsforschungszentrum (DKFZ), Heidelberg, Germany, 2Heidelberg University Hospital, Heidelberg, Germany, 3NCT Trial Center, Heidelberg, Germany, 4Charité – University Hospital, Berlin, Germany, 5Düsseldorf University Hospital, Düsseldorf, Germany 1

Introduction: Relapses from high-risk tumors, including several solid tumor entities and also hematopoietic malignancies, pose a major clinical challenge in pediatric oncology. The German INFORM registry study (INdividualized therapy FOr Relapsed Malignancies in children) addresses this problem using integrated next-generation sequencing to rapidly identify patient-specific potential therapeutic targets. Methods: Whole-exome, low-coverage whole-genome and RNA sequencing is complemented with microarray-based expression and DNA methylation profiling. Identified alterations are discussed and prioritized according to biological significance and potential druggability in a weekly molecular tumor board. Results: To date, 182 tumors have been profiled, with acute lymphoblastic leukemia accounting for 5% of these (n = 9). Turnaround time from tissue arrival to molecular results was 21 days on average. For patients post stem cell transplantation, we established an analysis pipeline to eliminate donor germline variant ‘contamination’ and generate reliable results within the same timeframe. In the total cohort, we detected one or more high-confidence druggable alterations in 118/182 (65%) patients. This ratio was higher for ALL-patients, where we identified druggable alterations in 9/9 (100%). Tyrosine kinases, the PI3K/mTOR pathway, MAPK pathway, and cell-cycle regulators were commonly affected. In one ALL patient with multiple relapses who had already undergone two stem cell transplantations, we identified a very rare INPP5D(SHIP1):ABL1 fusion. Treatment with tyrosine kinase inhibitors (dasatinib, later switched to ponatinib) combined with a multimodal immunotherapeutic treatment strategy lead to the induction of a complete remission for approximately 9 months.

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Conclusion: In summary, comprehensive profiling of pediatric tumors is feasible in a clinically meaningful timeframe, and provides valuable diagnostic information and potential therapeutic targets. We now aim to expand this approach to early-phase biomarker driven interventional trials. The relatively low number of ALL patients included so far, compared with the disease prevalence, may partly be explained by the high diversity of alternative diagnostic and therapeutic approaches. We are therefore establishing collaborations, e.g. with the iVAC-ALL peptide vaccination study, to maximize the available biological information and identify the optimal therapy concept for every individual patient. Disclosure: No conflict of interest disclosed. V79

CAR-T cell strategy – hype or hope? Rössig C.1 Universitätsklinikum Münster, Klinik für Kinderheilkunde und Jugendmedizin – Pädiatrische Hämatologie und Onkologie -, Münster, Germany 1

Chimeric antigen receptors (CARs) have recently emerged as a powerful means of redirecting T-cell functions toward B-lineage leukemias. CARs combine the ability of antibodies to recognize leukemia-associated surface antigens with the capacity of (co)stimulatory signal domains to activate T cells. Clinical trials now clearly demonstrate the potency of CAR-expressing T cells to eliminate human cancers. In a large proportion of patients with refractory CD19-expressing leukemias, T cells that express a CD19-specific CAR by genetic modification efficiently induced complete and even molecular remissions. The most significant toxicity of CAR T cell therapy is cytokine release syndrome that can be immediately life-threatening but has become manageable in most cases. The strength of CAR T cell therapy compared to bispecific T cell engagers is the ability of CAR gene-engineered T cells to persist in vivo and establish protective tumor-antigen specific memory responses. Indeed, CAR T cells were detected in the peripheral blood of many treated patients for several years, resulting in durable remissions at least in some patients. Another advantage is effective CNS penetration of CAR T cells and the ability to eliminate leukemic cells from this extramedullary compartment. On the other hand, a major challenge to broad implementation of the strategy in B cell cancers remains the need for individualized manufacturing of the T cell products. A drawback shared with other CD19 targeting strategies is the emergence of CD19-negative resistant clones by either CD19 splice variants or outgrowth of CD19-negative myeloid subclones under the selective pressure of CD19-directed CAR T cell therapy which has caused relapses in around 20% of acute lymphoblastic leukemia patients. The identification of additional CAR target antigens is a critical step also for extending the promise of this immunotherapeutic approach to hematological malignancies not derived from the B cell lineage, such as AML, and to solid cancers. A critical barrier against the use of engineered T cells for the treatment of bulky lymphomas and solid tumors is the tumor microenvironment which prevents both the recruitment and functional activity of therapeutic T cells. To overcome local immunosuppression and remain active in the tumor niche, CAR T cells may have to be administered in combination with agents that block the relevant inhibitory immune pathways. Disclosure: No conflict of interest disclosed.

Fortbildung

Hirnmetastasen und ZNS-Lymphome V80

Targeted-Therapy of brain metastasis of NSCLC, malignant melanoma and breast cancer Pukrop T.1 Universitätsklinikum Regensburg, Innere Medizin III, Regensburg, Germany

In the last years, we were pleased to achieve a significant improvement in the targeted treatment of metastatic tumor diseases, in particular in breast cancer, NSCLC and malignant melanoma. However, parallel to this positive development we observed an increase in CNS metastases. In contrast to the treatment of metastases in other organs, the body of evidence of systemic therapy in CNS-metastasis is sparse. One reason for this was the discussion about whether the blood-brain-barrier (BBB) is intact or not within CNS metastases. Today we know that the blood-brain barrier is disrupted and systemic therapy can be effective. For example, many TKI therapies have demonstrated a significant response even with CNS metastases, including meningeal metastasis. Thus, nowadays we have to consider systemic therapy for brain metastases not only in the context of clinical trials, but also in everyday clinical practice. For this reason it is also necessary and important to develop decision algorithms and clinical trials for these critically ill patients. Disclosure: Tobias Pukrop: Financing of Scientific Research: MSD, Lilly, Mundipharma; Other Financial Relationships: Reisefinanzierungen von Amgen und Mundipharma V82

Targeted therapy in CNS lymphoma Korfel A.1 Charité – Universitätsmedizin Berlin, Hämatologie, Onkologie und Tumorimmunologie, Berlin, Germany 1

Introduction: Despite some progress made in management of CNS lymphoma (CNSL) its prognosis remains worse as compared to other lymphoma localization. Moreover, toxicity, including both acute side-effects and delayed CNS toxicity, represents a major problem, particularly in the elderly and after combined radio-/chemotherapy. Thus, there is a need for development of novel drugs targeting key survival pathways in CNSL. Methods: English language literature has been reviewed for clinical studies with target therapy in CNS lymphoma. Results: CNSL are mostly aggressive lymphomas. While secondary CNS lymphoma remain to be nearly characterized, primary CNS lymphomas (PCNSL) are usually diffuse-large B-cell lymphomas exhibiting an activated B-cell like immunophenotype with ongoing immunoglobulin gene somatic hypermutation, Candidate targets and pathways identified thus far include the NFkB pathway, the B-cell receptor, the JAK/STAT pathway, IRF4, BCL-6 and PIM kinases as well chemokine pathways, macrophages and T-cells in tumor microeviroment. A phase II study of the mTOR inhibitor temsirolimus in relapsed/refractory PCNSL is the first completed and published trial on a target drug in PCNSL. A relatively high response-rate of 54% was found in the 37 patients treated, however, most responses were short-lived. In ongoing studies, the B-cell receptor inhibitor ibrutinib, the IRF/MUM1inhibitor lenalidomide, the dual PI3K/ mTOR inhibitor PQR309 and the check-point inhibitor nivolumab are being tested. Conclusions: A better definition of therapeutic targets will probably allow the development of new drugs to improve the outcome in CNSL. Disclosure: No conflict of interest disclosed.

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Fortbildung Thrombophilie

Freier Vortrag Zellbiologie

V85

V86

Perioperative anticoagulation: Is heparin bridging still relevant?

Influence of the JAK inhibitor Ruxolitinib on the migration process of dendritic cells (DC)

Angelillo-Scherrer A.1

Wolf D.1, Heine A.2, Quast T.3, Kolanus W.3, Trebicka J.4, Brossart P.2, Rudolph J.2

Universitätsklinik für Hämatologie und hämatologisches Zentrallabor / Inselspital, Bern, Switzerland 1

Background: More than 6 million Europeans are on long-term oral anticoagulation (OAC) therapy and about 10% of them require annually anticoagulation discontinuation for procedures. The approval of direct oral anticoagulants (DOAC) and novel data on perioperative anticoagulation with heparins render the management of perioperative anticoagulation therapy complex. Method: Relevant articles were selected in PubMed according to abstract content. To supplement the search, citations in pertinent review articles were examined. Current guidelines were studied. Results: Observational studies, large meta-analyses and a recent large randomized trial (the BRIDGE trial, NEJM 2015, 373: 823-33) revealed significant perioperative bleedings without reduction of thromboembolic events (TE) when heparin bridging is prescribed. These data point to an unbalanced effect of bridging toward bleeding. However, despite these evidences, bridging remains a common practice. Updated clinical guidelines suggest that heparin bridging is unnecessary for patients anticoagulated with DOACs. Heparin bridging should be considered for patients on longterm therapy with vitamin K-antagonists (VKA) temporarily interrupted in the periprocedural context if these patients are at highest risk of TE (≥ 10% per year). The decision of heparin bridging in these patients should be carefully weighted in presence of a concomitant high bleeding risk. Conversely, patients at low risk for TE should not be bridged. Interestingly, evidences from the literature suggest that patients with intermediate or unclear risks of TE or bleeding display higher perioperative bleedings than TE rates. Consequently, the individual patient and the procedure/ surgery need to be very well considered before the decision to bridge with heparin is made. Finally, the management of patients under VKA or DOAC therapy with prothrombin complex concentrates and novel antidotes specific for DOACs in the context of an urgent procedure or surgery will be discussed. This review will be illustrated by cases from the daily clinical practice. Conclusion: Scientific evidence supports the current guidelines and treatment algorithms for perioperative anticoagulation. Heparin bridging is proposed for a limited group of patients. Larger-scale studies are however still needed to determine the optimal periprocedural management of patients receiving DOAC therapy. Disclosure: No conflict of interest disclosed.

Universitätsklinikum Bonn, Bonn, Germany, 2Department of Internal Medicine III, Oncology, Hematology and Rheumatology, University Clinic Bonn (UKB), Bonn, Germany, 3Molecular Immunology & Cell Biology, Life and Medical Sciences Institute (LIMES), University of Bonn, Bonn, Germany, 4Department of Medicine I, Gastroenterology and Hepatology, University Clinic Bonn (UKB), Bonn, Germany 1

Introduction: Ruxolitinib is a potent JAK inhibitor approved for treatment of MPN and with a potential also for acute and chronic GvHD therapy. Ruxolitinib modified various immune cells including DC, that are essential antigen-presenting cells migrating via afferent lymphatics into T cell areas of draining lymph nodes (LN) for T cell priming. This process is guided by chemokine gradients and requires cytoskeletal rearrangement allowing cell movement. The aim of this study was a detailed analysis of the impact of ruxolitinib on DC migration with a particular focus on short-term effects of the drug and the identification of potential target molecules mediating these effects. Methods: Migration of human monocyte-derived DCs (moDC) or murine bone marrow-derived DCs (bmDCs) was analyzed in Transwell assays or dynamically by time-lapse microscopy within three dimensional collagen gels towards CCL-19 gradients. Morphological changes and signaling events were analyzed by immunofluorescent staining and Western Blot for changes in phosphorylation levels, respectively. siRNAs were used to knockdown talin, JAK1 or JAK2. Results: 2D migration of ruxolitinib-exposed DC is dose-dependently reduced in vitro. Analysis of the migratory phenotype of moDCs within 3D collagen gels revealed that ruxolitinib-exposed DCs are still able to sense chemokine gradients and form lamellipodia at the leading edge of the cell, whereas uropod retraction is inhibited. siRNA knockdown experiments revealed that this inhibitory effect is JAK1- and JAK2-independent. Moreover, ruxoltinib-mediated inhibition of migration is independent of integrin expression or signalling, as ruxolitinib-exposed DC express unaltered integrin levels and migration of talin 1-deficient DC (which are integrin decoupled) is also efficiently inhibited by ruxolitinib. On a molecular level we could show a reduced phosphorylation of the Rho-associated protein kinase (ROCK) in ruxolitinib-treated moDC upon CCL-19 stimulation. Finally, the observed migration phenotype induced by ruxolitinib could be mimicked by the ROCK inhibitor Y-27632. Conclusions: RhoA family members control important steps of cell migration, such as protrusion formation and consequently cell polarization. ROCK is a downstream effector of RhoA and stabilizes the actin cytoskeleton and acto-myosin II contraction. The observed reduction of ROCK phosphorylation may reveal an important mechanism of ruxolitinib-induced inhibition of DC migration Disclosure: Dominik Wolf: Advisory Role: Bexalta, Novartis; Financing of Scientific Research: Bexalta, Novartis; Expert Testimony: Bexalta, Novartis Janna Rudolph: No conflict of interest disclosed.

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

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V87

APC/CCdh1 controls differentiation and self-renewal in normal and malignant hematopoietic cells Ewerth D.1, Kreutmair S.1, Wider D.1, Felthaus J.1, Schüler J.2, Schmidts A.1, Duyster J.1, Illert A.-L.1, Engelhardt M.1, Wäsch R.1 Klinik für Innere Medizin I, Hämatologie, Onkologie und Stammzelltransplantation, Universitätsklinikum Freiburg, Freiburg, Germany, 2 Oncotest GmbH, Freiburg, Germany 1

Introduction: The balance between differentiation and self-renewal in hematopoietic stem and progenitor cells (HSPCs) is crucial and deregulation may lead to malignant transformation. The differentiation is predominantly initiated in G1 phase of the cell cycle when the E3 ligase anaphase-promoting complex or cyclosome (APC/C) is highly active. In association with its coactivator Cdh1 the APC marks proteins for proteasomal degradation. There is growing evidence that Cdh1 is a regulator of differentiation which has already been demonstrated in neurons, muscle cells or osteoblasts. Methods: Human CD34+ cells were collected from peripheral blood of G-CSF mobilized donors and cultured in presence of different cytokine combinations. The knockdown of Cdh1 was achieved by lentiviral delivery of shRNA into target cells. Transduced CD34+ cells were used for in vitro differentiation in liquid culture or CFU assay. For in vivo experiments purified cells were transplanted into NSG mice. The differentiation of AML cell line HL-60 with or without Cdh1 knockdown was induced by ATRA or PMA. Results: A detailed analysis of different subsets from both in vitro differentiated CD34+ cells and BM showed high Cdh1 level in CD34+ cells and low expression in myeloid or lymphoid cells. The depletion of Cdh1 (Cdh1-kd) had no effect on proliferation detected by CFSE and by measuring the cell cycle length of single cells. However, Cdh1-kd cells showed a significant maintenance of CD34+ cells under self-renewal conditions and during erythropoiesis with lower frequency of glycophorin A+ cells. In CFU assays, the Cdh1-kd resulted in less primary colony formation, notably CFU-GM and BFU-E, but significantly more secondary colonies. In a xenotransplant mouse model, human Cdh1-depleted CD34+ cells engrafted to a much higher degree in the murine BM 8 and 12 weeks after injection as shown by higher frequencies of human CD45+ cells. Moreover, we also found increased frequencies of human CD19+ B cells after transplantation of CD34+ Cdh1-kd cells. Primary AML blasts showed a significant downregulation of Cdh1 expression compared to normal CD34+ cells. The Cdh1-kd in AML cell line HL-60 led to less differentiated cells and a delay in PMA- and ATRA-induced differentiation. Conclusions: Loss of the APC/C coactivator Cdh1 supports self-renewal of CD34+ cells in vitro, facilitates engraftment capacity and B cell development of human HSPCs in vivo and contributes to the differentiation block in AML. Disclosure: No conflict of interest disclosed. V88

NAMPT-NAD+-SIRT2-triggered deacetylation of LMO2 protein is indispensable in the early hematopoiesis through TAL1 complex formation Morishima T.1, Lindner C.1, Bernhard R.1, Zahabi A.1, Dannenmann B.1, Nasri M.1, Samareh B.1, Kanz L.1, Welte K.1, Skokowa J.1 Universitätsklinikum Tübingen, Tübingen, Germany

Introduction: Nicotinamide phosphoribosyltransferase (NAMPT)- adenine dinucleotide (NAD+)- sirtuins (SIRTs) pathways regulate transcriptions through protein deacetylase function of SIRTs. In this study, we analyzed the roles of this pathway in early hematopoiesis. Methods: We tested NAMPT- or SIRT-inhibitors in human induced pluripotent stem (iPS) cells hematopoietic differentiation model. Results: We differentiated iPS cells with the specific inhibitor of NAMPT and found marked reduction of VEGFR2+CD34+ early hematopoietic progenitor cells at day 6. As qRT-PCR results shows SIRT2 mRNA expression

Oncol Res Treat 2016;39(suppl 3):1–104

is increasing in this stage, we tested SIRT2 specific inhibitor and found significant reduction of these progenitor cells. Furthermore, the emergence of CD43+ hematopoietic progenitor cells and colony forming ability at day 13 were completely suppressed by inhibition of SIRT2. These results indicate that NAMPT-NAD+-SIRT2 signaling is needed for early hematopoiesis. To evaluate downstream targets, we analyzed mRNA and protein expressions of early hematopoietic genes (GATA2, RUNX1, LMO2, and TAL1) on day 6 and found that they were not down-regulated by NAMPT or SIRT2 inhibition. With the idea of post-transcriptional modifications, we quantified acetylation of these proteins in the cells treated or not with SIRT2 inhibitor and found that only LMO2 was deacetylated by SIRT2. Interaction between SIRT2 and LMO2 was confirmed with Duolink and co-immunoprecipitation. LMO2 is known as a component of TAL1 complex, which consist of LMO2, TAL1, LDB1, E47 and GATA1/2 proteins and has transcriptional activity as a complex on hematopoiesis-specific target genes. Interestingly, mRNA expressions of all these target genes were markedly downregulated by inhibition of NAMPT or SIRT2. We made acetylation- (KQ) and deacetylation- (KR) mimic mutants of LMO2 protein at K74 and K78 lysine residues and compared LMO2-LDB1 interaction by co-immunoprecipitation. This experiment revealed that KQ-mutant had impaired interaction in comparison to WT and KR-mutant. These results indicate that deacetylation of LMO2 protein by SIRT2 is required for the interaction between LMO2 and LDB1 proteins. Conclusions: NAMPT-NAD+-SIRT2 pathway plays an indispensable role in the early hematopoiesis by deacetylation of LMO2 protein which enables the interaction between LMO2 and LDB1 proteins and complete TAL1 complex formation. Disclosure: No conflict of interest disclosed. V89

Unaffected myeloid differentiation of iPS cells derived form a cyclic neutropenia (CyN) patient with ELANE mutation Zahabi A.1, Morishima T.1, Bernhard R.1, Kanz L.1, Welte K.2, Skokowa J.1 University Hospital Tuebingen, Oncology, Hematology, Immunology, Rheumatology and Pulmonology, Tübingen, Germany, 2University Children’s Hospital, Tübingen, Germany 1

Background: Cyclic neutropenia (CyN) is a hematologic disorder in which blood cell counts particularly granulocytic neutrophils, monocytes, platelets, and reticulocyte numbers show cycles at appr. 21 day intervals. The majority of CyN patients (ca 90%) harbor inherited mutations in the ELANE gene. Intriguingly, same ELANE mutations are present in two different bone marrow failure syndromes: congenital as well as in cyclic neutropenias. It is unclear how mutation in the same gene cause congenital (CN) or cyclic neutropenia (CyN). The pathomechanism of cycling hematopoiesis downstream of ELANE mutations is also unclear. Recent studies using inducible pluripotent stem cells (iPSCs) derived from severe congenital neutropenia (CN) patients harbouring ELANE mutations demonstrated markedly diminished granulocytic differentiation of these cells in vitro (Nayak RC, et al. JCI 2015; Hiramoto T, et al. PNAS 2013). Interestingly, correction of ELANE gene mutation in iPSCs from a CN patient using CRISPR/Cas9 technology restored defective granulopoiesis, suggesting the monogenic origin of the ELANE mutation caused congenital neutropenia (Nayak RC, et al. JCI 2015). These data suggest that additional gene mutations or epigenetic defects in combination with mutated ELANE might be responsible for the pathogenesis of cyclic neutropenia. Aims: In the present study we evaluated the in vitro myeloid differentiation of iPSCs derived from the CyN patient harboring sporadic heterozygous ELANE mutation (c.761C>G p.W241L), in comparison to iPSCs derived from healthy individuals. Methods: We used embryoid-body (EB)-based protocol of granulocytic differentiation from iPSCs described by N. Lachmann et al. (Stem Cell Reports 2015). Results: Interestingly, we found that myeloid differentiation of iPSCs derived from CyN patient, was comparable to that of healthy individual iPSCs, as revealed by the analysis of the percentage and absolute numbers of

Abstracts

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CD45+/CD11b+, CD45+/CD16+ and CD45+/CD15+ cells as well as by the examinations of the cell morphology on cytospin preparations. Summary/ Conclusion: These data suggest that the in vitro hematopoiesis in CyN is normal as judged by iPSC technology and that in vivo additional humoral or bone marrow niche components might influence myeloid cell cycling in CyN. Disclosure: No conflict of interest disclosed. V90

Differential use of metabolic substrates during erythropoiesis and granulopoiesis Böhme C.1,2, Billing C.1, Walker M.3, Noack N.1, Pompe T.2, Niederwieser D.1, Whetton A.3, Cross M.1 Universitätsklinikum Leipzig AöR, Selbständige Abteilung für Hämatologie/ int. Onkologie, Leipzig, Germany, 2Universität Leipzig, Institut für Biochemie, Leipzig, Germany, 3University of Manchester, Stem Cell Proteomics Laboratory, Manchester, United Kingdom 1

Introduction: Erythrocytes and neutrophilic granulocytes are constantly produced in large numbers from a common progenitor in the same region of haematopoietic marrow. We are interested in the possibility that these lineages use different patterns of metabolic activity to fuel the high levels of proliferation required. Materials and methods: CD34+ progenitors were purified from umbilical cord blood by MACS. Following short term culture, CD34+ CD133+ (multipotent and lympho-myeloid progenitors, MPP / LMPP) and CD34+ CD133low (erythro-myeloid progenitors, EMP) were purified by FACS. The metabolic requirements of colony-formation were assessed in a system developed to allow the exchange of media without disturbing the cells. After 14 days, cells were recovered from the methylcellulose for FACS and PCR analysis. Metabolite exchange with the medium during erythropoiesis and granulopoiesis was assessed in liquid cultures of murine FDCP-Mix cells. Mitochondria purified from the same cells were used in proteomic analysis (iTRAQ). Results: Even in the absence of detectable glucose, regular exchange of the medium over the first 3 days of culture was sufficient to generate robust colony formation over 14 days, suggesting that at least a subpopulation of progenitor cells is capable of gluconeogenesis. Consistent with this, the expression of both PEPCK 1 and 2 was detected in colonies. Colony formation from all populations tested required glutamine. However, high glutamine concentrations selectively blocked the development of erythroid colonies from EMPs. Similarly, FDCP-Mix cells undergoing either granulocytic or erythroid differentiation take up glutamine from the medium, but large amounts of glutamate are released back into the medium only under erythroid conditions. This suggests a block in glutaminolysis specifically in the erythroid lineage. A proteomic analysis of mitochondria from FDCP-Mix cells differentiating along each lineage showed erythroid differentiation to be accompanied down-regulation of Gpt2, an amino transferase responsible for channelling glutamate into the TCA cycle. Discussion: The novel colony assay procedure has identified previously unknown features of haematopoietic metabolism, consistent with the lineage-specific use of limiting substrates. Metabolic compartmentalisation may help to make maximal use of limiting substrates and/or provide a level of metabolic feedback between the lineages. Disclosure: No conflict of interest disclosed.

V91

Immunosuppressive capacity of mesenchymal stromal cells depends on allogeneic donor T-cells and correlates with metabolic activity Nold P.1, Killer M.1, Henkenius K.1, Fritz L.1, Hackstein H.2, Neubauer A.1, Brendel C.1 Universitätsklinikum Gießen und Marburg GmbH, Hämatologie/Onkologie/ Immunologie, Marburg, Germany, 2Universitätsklinikum Gießen und Marburg GmbH, Zentrum für Transfusionsmedizin und Hämotherapie, Giessen, Germany 1

Introduction: Mesenchymal stromal cells (MSC) have entered the clinic as Advanced Therapy Medicinal Product (ATMP) and are currently evaluated in a wide range of studies for tissue regeneration or in autoimmune disorders. Various efforts have been made to standardize and optimize expansion and manufacturing processes but until now reliable potency assays reflecting therapeutic effectiveness of MSC are still lacking. As recent findings suggest superior therapeutic efficacy of freshly administered MSC in comparison to frozen cells we sought to determine the immunosuppression capacity of MSC in relation to their metabolic activity. Methods: Human MSC were obtained from bone fragments of patients and were employed in co-culture with peripheral blood mononuclear cells (PBMC) in an allogeneic T-cell proliferation assay to measure immunosuppressive function. Metabolic activity of MSC was measured real-time as aerobic glycolysis quantified by the extracellular acidification rate (ECAR) and mitochondrial respiration quantified by the oxygen consumption rate (OCR). Results: We show that MSC induced suppression of T-cell proliferation varied enormously between different healthy donors of PBMC. Moreover, direct contact of PBMC and MSC increased the metabolic activity of MSC in a PBMC donor dependent manner. Enhanced lactate production as a surrogate parameter for glycolysis and increased oxygen consumption was paralleled by higher T-cell suppression capacities of MSC. The cryopreservative dimethylsulfoxide decreased metabolic and immunosuppressive performance of MSC while valproic acid enhanced metabolism and T-cell suppression. Conclusion: Metabolic and immunosuppressive capacity of MSC are linked closely but are highly dependent on the predisposition of interacting T-cells which enhance MSC metabolism upon direct contact in co-culture. Hence functional fitness and quality of MSC can be determined by measuring metabolic activity and can be enhanced by exposure to valproic acid. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Infektionen / supportive Therapie V92

Detection and characterization of Aspergillus fumigatus azole resistance and cyp51A combinations of mutations in clinical isolates and primary clinical samples of hematological patients – preliminary results comparing two molecular assays Spiess B.1, Postina P.1, Boch T.1, Miethke T.2, Dietz A.2, Merker N.1, Hofmann W.K.1, Buchheidt D.1 Universitätsmedizin Mannheim; III. Medizinische Klinik, Mannheim, Germany, Universitätsmedizin Mannheim; Institut für Medizinische Mikrobiologie und Hygiene, Mannheim, Germany 1 2

Introduction: In hematological patients, there is a rising incidence of invasive aspergillosis (IA) caused by azole resistant Aspergillus fumigatus (A. fumigatus) that reveals a combination of novel mutations. As the diagnosis of IA is rarely based on positive culture yield in this group of patients, molecular detection of resistance mutations directly from clinical samples is crucial.

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

CONTENTS AUTHOR INDEX

Methods: In addition to our established azole resistance PCR assays detecting the frequent mutation combinations TR34/L98H/M220I and TR46/Y121F/T289A in the A. fumigatus cyp51A gene, we investigated in parallel the commercially available AsperGenius® real time PCR detection system (PathoNostics BV, Maastricht, The Netherlands) detecting the cyp51A alterations TR34/L98H and Y121F/T289A directly from clinical samples and isolates. We analyzed phenol-chloroform extracted DNA aliquots and compared both PCR methods, not the preceding different DNA extraction protocols. Results: Our novel Y121F and T289A PCR assays showed a detection threshold of 300 fg genomic A. fumigatus DNA. As positive control for the feasibility of these assays, we detected the TR46/Y121F/T289A mutation combination in two isolates of hematological patients with known cyp51A alterations and in a lung biopsy sample of a patient with AML and proven IA. Results comparing the two test systems are shown in table 1. Tab. 1. Resistance mutations (ARAF=azole resistant A. fumigatus).

DNA from Isolate 1 Isolate 2 Clinical samples Lung biopsy 1 Lung biopsy 2 Brain biopsy BAL

Mannheim ARAF PCRs TR46/Y121F/T289A TR46/Y121F/T289A TR46/Y121F/T289A L98H L98H/TR34 L98H/TR34

AsperGenius® PCRs Y121F/T289A Y121F/T289A negative negative negative L98H/TR34

Conclusions: In order to detect A. fumigatus azole resistance directly from clinical samples, we broadened the spectrum of our PCR assays. The combination of our results with these of the AsperGenius® assay revealed that our conventional PCR assays are more suitable for the analysis of biopsy samples. Negative results of the AsperGenius® assay in biopsy samples could most likely be due to different PCR conditions and the interference of the PCR reactions caused by high amounts of human DNA. Despite this, we consider non-culture based molecular detection of Aspergillus azole resistance to be of high epidemiological and clinical relevance. Disclosure: Birgit Spiess: No conflict of interest disclosed. Dieter Buchheidt: Advisory Role: Basilea, Gilead Sciences; Financing of Scientific Research: Astellas, Gilead Sciences, Merk Sharp & Dohme/Merck, Pfizer; Expert Testimony: Gilead Sciences, Pfizer V93

Peptide vaccination against cytomegalovirus (CMV) can clear the virus load after allogeneic stem cell transplantation even from a CMV seronegative donor Schmitt M.1, Schmitt A.1, Wiesneth M.2, Hückelhoven A.1, Wu Z.3, Kuball J.4, Wang L.1, Schauwecker P.2, Hofmann S.5, Götz M.5, Michels B.1, Maccari B.2, Wuchter P.1, Mertens T.3, Schnitzler P.6, Döhner H.5, Ho A.D.1, Bunjes D.W.5, Dreger P.1, Schrezenmeier H.2, Greiner J.5,7 Medizinische Klinik V, Universitätsklinikum Heidelberg, Heidelberg, Germany, Institut für Klinische Transfusionsmedizin und Immungenetik Ulm (IKT), Ulm, Germany, 3Institut für Virologie, Universitätsklinikum Ulm, Ulm, Germany, 4 Department of Hematology, University Medical Center, Utrecht, Netherlands, 5 Klinik für Innere Medizin III, Universitätsklinikum Ulm, Ulm, Germany, 6 Department für Infektiologie, Virologie, Universitätsklinikum Heidelberg, Heidelberg, Germany, 7Medizinische Klinik, Diakonie-Klinikum Stuttgart, Stuttgart, Germany 1 2

Introduction: Cytomegalovirus (CMV) reactivation occurs particularly in patients receiving an allogeneic stem cell graft from a seronegative donor. CMV reactivation is associated with a high risk of disease and mortality. The nonamer peptide NLVPMVATV derived from CMV phosphoprotein 65(CMVpp65) is highly immunogenic. Here we report on a clinical phase I peptide vaccination trial with this peptide in a water-in-oil emulsion (Montanide™) plus administration of granulocyte-macrophage colony stimulating factor (GM-CSF) (EudraCT number: 2010-018884-40).

Oncol Res Treat 2016;39(suppl 3):1–104

Methods: Four vaccines were administred subcutaneously at a biweekly interval to ten patients. Clinical course and CMVpp65 antigenemia were monitored. CMV-specific CD8+ and gamma-delta T cells were analyzed by multi-color flow cytometry. We established a neutralizing anti-CMV antibody assay and correlated it to clinical parameters. Results: Peptide vaccination was well tolerated and no drug-related serious adverse events were detected. Seven of nine patients with CMVpp65 antigenemia cleared the CMV after four vaccinations and are still free from viremia until now. Two patients with CMV reactivation showed no clinical response, i.e. persisting CMV antigenemia. One patient received prophylactic vaccination and did not develop viremia. In four patients an increase in frequency of both CMV specific CD8+ T cells Vdelta2-negative gamma-delta T cells was detected by factor 6 and 7, respectively. In four patients also titers of neutralizing antibodies increased up to tenfold over the time of vaccination. Humoral and cellular immune responses correlated with clearance of the CMV load in the patients. Conclusion: In patients after allogeneic stem cell transplantation with high risk for CMV reactivation, we demonstrated that administration of CMVpp65 peptide vaccination was safe, well tolerated and clinically encouraging. Further studies with larger patient cohorts are planned. A study with prophylactic CMVpp65 peptide vaccination is ongoing in patients receiving a kidney transplantation. Disclosure: No conflict of interest disclosed. V94

Different localization of Dectin-1 isoforms governs its recognition of ß-glucan and signaling quality Fischer M.1, Müller J.P.2, Spies-Weisshart B.1, Gräfe C.1, Hochhaus A.1, Scholl S.1, Schnetzke U.1 Universitätsklinikum Jena, Klinik für Innere Medizin II, Abteilung Hämatologie und Internistische Onkologie, Jena, Germany, 2Universitätsklinikum Jena, Institut für Molekulare Zellbiologie, CMB, Jena, Germany 1

Introduction: The incidence of invasive fungal infections (IFI) in hematological patients has increased substantially during the last decades. Thus, there is a need for a better understanding of antifungal immunity. Dectin-1, a C-type-lectin-like receptor, is recognized as a major pattern-recognition receptor for fungal ß-glucans and contributes to anti-fungal immunity. The two major human isoforms A and B are expressed on human monocytes as well as on macrophages and dendritic cells. Both proteins share the same carbohydrate recognition domain while isoform A is characterized by an additional stalk region and its N-linked glycosylation site. Methods: Expression of Dectin-1 isoform A and B was analyzed in monocyte-derived cells. Cell lines (THP-1, HEK293) stably expressing Dectin-1 isoform A or B were generated by retroviral transduction. Subcellular localization of Dectin-1 was analyzed using flow cytometry and confocal laser scanning microscopy (CLSM). Furthermore, binding capacity of ß-glucan was examined by flow cytometry and signaling quality using immunoblotting. Results: Cell line models demonstrate a ten times higher surface membrane expression of isoform A compared with B, despite comparable mRNA und whole cell protein levels. CLSM imaging validates an intracellular retention of isoform B on protein level. The reduced surface membrane expression of isoform B correlates with a reduced ß-glucan binding capacity and an impairment of Syk and p38 dependent signaling quality. Inhibition of N-linked glycosylation of isoform A by tunicamycin leads to an equalization of surface membrane expression comparable to isoform B. Conclusions: This work reveals the substantial role of N-linked glycosylation of Dectin-1 and its impaction on surface membrane expression and consequently ß-glucan mediated signal transduction. Different expression profiles of human Dectin-1 isoforms on monocyte-derived cells may be associated with cell-type-dependent effector mechanisms of antifungal immunity. Disclosure: Mike Fischer: No conflict of interest disclosed. Ulf Schnetzke: Expert Testimony: Firma Gilead

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Selenium for the prevention and management of adverse effects of cancer treatments Renner P.1, Dennert G.2, Kalisch A.3, Horneber M.3 Universitätsklinik für Innere Medizin 8, Paracelsus Medizinische Privatuniversität, Klinikum Nürnberg, Schwerpunkt Kardiologie, Nürnberg, Germany, 2Angewandte Sozialwissenschaften, Fachhochschule Dortmund, Sozialmedizin und Public Health mit Schwerpunkt Geschlecht und Diversität, Dortmund, Germany, 3Universitätsklinik für Innere Medizin 5, Paracelsus Medizinische Privatuniversität, Klinikum Nürnberg, Schwerpunkt Onkologie/ Hämatologie, Nürnberg, Germany 1

Introduction: This is an updated version of the original Cochrane review published in 2009. Selenium is an essential trace element and is involved in antioxidant protection and the redox-regulation in humans. It has been claimed for its efficacy in supportive cancer care. Selenium supplements are frequently used by cancer patients during anticancer treatments. The aim of this systematic review was to evaluate the efficacy of selenium for the prevention and management of adverse effects of chemotherapy or radiotherapy and for after-effects of tumor surgery. Methods: For this update we searched electronic databases, including those of Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE and EMBASE, up to September 2015. We included randomised controlled trials of selenium for the prevention or management of adverse effects of chemotherapy, radiotherapy and of after-effects of tumour surgery. At least two review authors independently determined suitability for inclusion and extracted and assessed study data. Results: We found four new studies with a total of 253 participants for this update. Together with the three previously identified studies, the update is now based on a total of seven studies with 411 participants suffering from various cancers at different stages. All studies used different preparations and doses of oral selenium to prevent a heterogenous set of side effects or toxicities. Therefore, no meaningful metaanalysis was possible. Conflicting evidence was found for protective effects of selenium on renal toxicity of cisplatin and other chemotherapeutic agents. Conflicting evidence was also found for effects of selenium to prevent oral mucositis during chemotherapy, chemoradiation or radiation therapy. Due to high risk of bias in the reporting studies, only low quality evidence was found for protective effects of selenium on either radiation-induced diarrhoea during pelvic radiotherapy, or the recurrence of erysipelas in lymphedematous skins after mastectomy or on postoperative lymphedema after oral surgery. Conclusions: In summary, we found no convincing evidence for beneficial effects of selenium for the prevention or management of chemotherapy- or radiotherapy-related adverse effects or for after-effects of tumor surgery in cancer patients. Four additional identified studies for this review did not change our conclusions. Funding: Ernst und Anita Bauer-Stiftung, Deutsche Krebshilfe Disclosure: No conflict of interest disclosed. V96

Glutamine in the prevention of tumortherapy-induced diarrhea Leithold C.1, Jahn F.1, Rüssel J.1, Unverzagt S.2, Müller-Tidow C.1, Jordan K.1 Universitätsklinikum Halle-Wittenberg, Halle, Germany, 2Institut für Medizinische Biometrie, Epidemiologie und Informatik, Halle, Germany 1

Introduction: Glutamine is a main energy resource for cells with high turnover rates, such as enterocytes or cells of the immune system. There are a lot of animal studies supporting the important role of glutamine in mucosal integrity and gastrointestinal function. Also numerous studies in human showed benefit in improving gastrointestinal function using glutamine as a supplement. Due to the presumed mode of action it is assumable that glutamine might be of benefit in preventing mucosal damage in patients receiving chemo- or radiotherapy. This systematic review assessed the effects of oral or parenteral glutamine in preventing diarrhea due to tumor therapy.

Abstracts

Methods: We conducted a systematic literature search using Medline (via Ovid and PubMed) and Cochrane Library to find randomized controlled trials which studied the efficacy of orally or parenterally administered glutamine for the prevention of diarrhea in patients receiving chemo- or radiotherapy. Duration, incidence and severity of diarrhea were documented as outcome measurements. We conducted subgroup analyses for chemo- and radiotherapy-induced duration of diarrhea and oral or parenteral application of glutamine. Results: We found 2891 records and identified a total of 14 studies that met the inclusion criteria, eight studies were finally included in meta-analysis. Neither oral (MD: -0.44 d; 95% CI: -1.56 d-0.68 d) nor parenteral administration of glutamine (MD: -0.68 d; 95% CI: -2.72 d-1.36 d) could significantly reduce duration of chemotherapy-induced diarrhea (CID). For the prevention of radiotherapy-induced diarrhea (RID) we found inconsistent study results suggesting that the administration of oral glutamine might even increase the incidence of diarrhea (OR: 13.03; 95% CI: 2.68-63.22). Conclusions: Glutamine is not effective to prevent duration or severity of CID or RID and has no impact on the incidence of CID or RID. Since study results for RID even described an increase of incidence of diarrhea, there is no point using glutamine as a preventive agent. These results are in contrast with former meta-analysis. The results of our systematic review formed the basis for the recommendation of the S3 guidelines of “Supportive therapy”. Disclosure: No conflict of interest disclosed. V97

Bisphosphonates for patients with bone metastases from prostate cancer. A Cochrane review update including more than 5,000 patients Macherey S.1, Jahn F.2, Jordan K.2, Skoetz N.1 Cochrane Haematological Malignancies Group, Klinik I für Innere Medizin, Universitätsklinik Köln, Köln, Germany, 2Klinik IV für Innere Medizin, Universitätsklinik Halle/Saale, Halle/Saale, Germany 1

Introduction: Patients with advanced prostate cancer frequently present with bone metastases. These patients are usually affected by bone pain, pathological fractures or spinal cord compression and often need interventions like surgery or radiation of the bone (skeletal-related events, SRE). Bisphosphonates are well established in the multimodal therapy of bone pain in patients with bone metastasis from prostate cancer. Only few studies have investigated the effect of bisphosphonates on overall survival (OS). Hence, we performed a meta-analysis aimed at the effect of bisphosphonates on the patients’ general outcome. Methods: In December 2015, we performed a literature search by screening the electronic databases MEDLINE and the Cochrane library or databases of ongoing clinical trials. Furthermore, we hand searched conference proceedings of the American Society of Clinical Oncology and the references of any identified trial. We defined pain response as primary end point and mortality, SRE, analgesic consumption, disease progression, renal adverse events and osteonecrosis of the jaw (ONJ) as secondary end points for the statistical analysis. The GRADE software was used to evaluate each trial’s quality of evidence. This study was partly funded by the German Cancer Aid (No. 110645). Results: A total of 21 randomized controlled trials reporting on approximately 5,000 patients could be included. Bisphosphonates probably improved the patients’ pain response (RR: 1.21; 95% CI: 0.99, 1.48; p = 0.05), reduced the number of skeletal-related events (RR: 0.90; 95% CI: 0.81, 0.99; p = 0.03) and the patients’ disease progression (RR: 0.94; 95% CI: 0.89, 0.99; p = 0.03) in comparison to the control group. There is no evidence for a difference between bisphosphonates and placebo considering mortality (RR: 0.98, 95% CI: 0.91, 1.05) or renal adverse events (RR: 1.04, 95% CI 0.76, 1.42). Bisphosphonates increase the rate of ONJ (RR: 6.77, 95% CI: 3.17, 14.46). It is uncertain whether bisphosphonates improve analgesic consumption (RR: 1.19, 95% CI: 0.87, 1.63) in those patients.

Oncol Res Treat 2016;39(suppl 3):1–104

CONTENTS AUTHOR INDEX

Conclusions: Bisphosphonates lead to a significant reduction of skeletal-related events and disease progression in patients with prostate cancer and bone metastases. The use of bisphosphonates showed an improvement in pain response, but is associated with a higher incidence for osteonecrosis of the jaw. There is no evidence for an effect of bisphosphonates on mortality or analgesic consumption. Disclosure: No conflict of interest disclosed.

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Interprofessionelle Sitzung: Geriatrische Onkologie (für Ärzte und Pflegekräfte) V100

Decision-making capacity in elderly cancer patient Weidmann-Hügle T.1 Institut für Biomedizinische Ethik und Medizingeschichte, Universität Zürich, Zürich, Switzerland 1

Patient autonomy is a fundamental and indispensable moral principle in contemporary medicine as well as in medical ethics. It is the basis of much theory and most regulation. The main legal process to promote patient autonomy is through informed consent. Within this process the patient is being comprehensively informed about a pending treatment. Informed consent involves discussion of the nature of a procedure, its possible risks and benefits, and alternative treatments. Ideally the patient then makes a free and non-pressured decision about further treatment, which is consistent with his values and life´s plan. Key to the legal validity of a patient’s consent (or dissent) is her decision-making capacity (in German: “Urteilsfähigkeit” or “Einwilligungsfähigkeit”). In general terms, the requirement of having decision-making capacity is then met, if a person is capable 1) of understanding and appreciating relevant information, 2) of processing this information, 3) of assessing possible consequences of available treatment options, and, finally, if she is capable of coming to a decision. Cancer occurs frequently among older adults. Cognitive impairment and depression are also common in this population. Both cognitive impairment and depression, coupled with stress, uncertainty, or side effects from treatment may have an impact on a person’s ability to make decisions. Decisions about cancer treatment can be demanding and require sufficient cognitive capacity, especially given the fact that there is often uncertainty in the outcomes and the decisions rely on weighing probabilities of side effects versus benefits Therefore, knowledge about approaches and instruments for the assessment of a patient’s decision-making capacity is crucial for healthcare professionals in oncology. Furthermore, healthcare professionals must be aware of the various factors which can impact decision-making capacity. In this presentation I will focus on the theoretical and legal concepts of decision-making capacity and factors influencing this, on assessment approaches, and on how nurses can support elderly cancer patients in the process of decision-making.

V101

Geriatric cancer patients- Comprehensive geriatric assessment and patient-reported quality of life Schmidt H.1, Boese S.1, Lampe K.2, Jung M.1, Jordan K.3, Fiedler E.4, Müller-Werdan U.5, Vordermark D.2 Martin Luther University Halle-Wittenberg, Institute of Health and Nursing Science, Halle (Saale), Germany, 2University Hospital Halle Saale, Department of Radiation Oncology, Halle (Saale), Germany, 3University Hospital Halle Saale, Department of Hematology and Oncology, Halle (Saale), Germany, 4University Hospital Halle Saale, Department of Dermatology, Halle (Saale), Germany, 5 Charite – Universitätsmedizin Berlin, Chair of Geriatrics, and Protestant Geriatric Centre, Berlin, Germany 1

Introduction: Oncologic therapy of elderly cancer patients is often complicated by comorbidities, reduced functioning and the organization of care at home. For long term care the maintenance of functionality and health related quality of life (HRQOL) is important. Therefore, prior to cancer specific therapy, the identification of relevant risk factors by comprehensive geriatric assessment (CGA) is recommended for elderly cancer patients. Methods: Aiming to maintain HRQOL we developed an interdisciplinary care program based on comprehensive geriatric assessment (CGA) and patient reported HRQOL comprising tailored supportive measures and telephone based counselling during 6 months aftercare. The intervention was pilot-tested in three centres at the University Hospital Halle Saale to examine feasibility, acceptance and potential benefit. Inclusion criteria: Oncologic patients >70 years with at least one comorbidity and/or one functional impairment with written informed consent. Primary endpoint is HRQOL (EORTC QLQ-C30, ELD14), measured at admission and 6 month-follow-up. First Results: Out of n = 226 eligible patients n = 100 participated (44%), mean age: 76.3 years (SD = 4.8), 47% female, comorbidities mean: n = 5 (SD = 2.8). Follow-up will be completed by July 2016. Preliminary analyses show large inter-individual differences regarding functional status, reported symptom intensity and supportive needs. Individualized supportive care was triggered by summarized individual results that were presented to the physicians in charge of the respective patients (e.g. malnutrition, reduced HRQOL, reduced physical functioning, high symptom-intensity and depression). Preliminary analyses for the primary endpoint global HRQOL (n = 46) showed clinical relevant improvement of HRQOL (≥10 pts.) for 35%, no change for 41% and worsening for 24%. Concurrent with worsening of global HRQOL we found a deterioration of physical function, mobility and fatigue. Further case based analyses comparing professional assessments and self-assessments including HRQOL will be presented in connection with follow-up data. Conclusion: First results show feasibility and potential usefulness of the combination of CGA and HRQOL to complement standard assessments and to decide on individualized therapeutic measures and after care. Disclosure: Heike Schmidt: No conflict of interest disclosed. Dirk Vordermark: Advisory Role: Roche, Boehringer, Bristol-Myers Squibb,; Financing of Scientific Research: Merck, Lilly

Disclosure: No conflict of interest disclosed.

Oncol Res Treat 2016;39(suppl 3):1–104

Abstracts

CONTENTS AUTHOR INDEX

Freier Vortrag

V104

Mastozytose, myeloproliferative Neoplasien

Mast cell leukemia: clinical heterogeneity, molecular aberrations and prognostic factors

V103

Jawhar M.1, Schwaab J.1, Meggendorfer M.2, Naumann N.1, Kluger S.1, Horny H.-P.3, Sotlar K.3, Haferlach T.2, Fabarius A.1, Hofmann W.-K.1, Reiter A.1, Metzgeroth G.1

Expression, regulation and function of the cell adhesion molecule CD44 in neoplastic mast cells and stem cells in patients with systemic mastocytosis Mueller N.1, Wicklein D.2, Eisenwort G.1,3, Boehm A.1,4, Herrmann H.1,3, Stefanzl G.1,3, Hoermann G.3,5, Sperr W.R.1,3, Arock M.6, Schumacher U.2, Valent P.1,3 Medical University of Vienna, Internal Medicine I, Division of Hematology and Hemostaseology, Vienna, Austria, 2University Medical-Center HamburgEppendorf, Institute of Anatomy and Experimental Morphology, University Cancer Center, Hamburg, Germany, 3Medical University of Vienna, Ludwig Boltzmann Cluster Oncology, Vienna, Austria, 4Elisabethinen Hospital, Linz, Austria, 5Medical University of Vienna, Department of Laboratory Medicine, Vienna, Austria, 6Ecole Normale Supérieure de Cachan, LBPA CNRS UMR8113, Paris, France 1

The Hermes antigen CD44 is a multifunctional adhesion molecule that plays an essential role in the homing and expansion of neoplastic stemand progenitor cells in various myeloid malignancies. Although mast cells (MC) are known to express CD44, little is known about the regulation and function of this receptor on neoplastic mast cells and stem cells in patients with systemic mastocytosis (SM). In the current study, we found that CD34+/CD38- stem cells, CD34+/ CD38+ progenitor cells, and KIT+/CD34- MC invariably express CD44 in all patients with indolent SM (ISM), SM with associated hematologic nonMC-disease (SM-AHNMD), aggressive SM (ASM), and MC leukemia (MCL). In addition, all human MC lines examined, including HMC-1.1, HMC-1.2, ROSAKIT wt, ROSAKIT D816V and MCPV were found to express cytoplasmic and cell surface CD44. We next examined the regulation of expression of CD44 in neoplastic MC. Incubation with the MEK inhibitor RDEA119 (0.1-5 µM) or the STAT5 blocker pimozide (2.5-10 µM) for 48 hours resulted in a significant, dose-dependent downregulation of surface expression of CD44 in all MC lines examined. By contrast, incubation with the demethylating agents decitabine (0.1-5 µM) or azacitidine (0.1-5 µM) for 96 hours resulted in a dose-dependent and significant upregulation of CD44 expression in all MC lines tested. We were also able to detect soluble CD44 in the sera of patients with ISM, SM-AHNMD, ASM, and MCL as well as in the supernatants of neoplastic MC lines. In order to define a functional role for CD44 expressed on neoplastic MC, we transduced shRNA against CD44 as well as a control shRNA into HMC-1.2 cells, and injected these cells into the skin of severe combined immunodeficient (SCID) mice. In this xenotransplantation model, the shRNA-mediated knockdown of CD44 was found to lead to reduced MC expansion, reduced tumor formation, delayed ulceration, and to a significantly prolonged survival compared to cells transduced with control shRNA. The formation of lung metastases, quantified by human Alu-sequence-specific qPCR, was found to decrease particularly (15-fold) in the CD44 knockdown group compared to control mice. Together, we show that CD44 is a relevant homing molecule expressed on neoplastic MC and CD34+ neoplastic stem- and progenitor cells in advanced SM. Our data also suggest that CD44 may serve as an interesting new target of therapy in patients with advanced SM.

University Medical Centre, Department of Hematology and Oncology, Mannheim, Germany, 2MLL Munich Leukemia Laboratory, Munich, Germany, 3 Ludwig-Maximilians-University, Institute of Pathology, Munich, Germany 1

Introduction: Mast cell leukemia (MCL) is a rare variant of systemic mastocytosis (SM). Data on disease characteristics, treatment, and prognosis are limited due to small patient numbers. Methods: Here, we report on 25 MCL patients (median age 67 years, range 45-82; male 64%). Results: An associated hematologic neoplasm (AHN), e.g. CMML (n = 7), MDS/MPNu (n = 6) or MDS (n = 4) was diagnosed in 17/25 (68%) patients. Primary MCL was found in 15/25 (60%) patients, and secondary MCL evolving from SM-AHN or aggressive SM (after median 20 months; range, 4-71), in 10/25 (40%) patients. Median percentage of MC in BM smears and trephine biopsies were 25% (range 20-95) and 60% (range, 20-60), respectively. MC in peripheral blood (PB) ≥10% (leukemic MCL) were only detected in 2/25 (8%) patients. Median serum tryptase level was 549 µg/L (range 160-1854). Hematologic C-findings such as hemoglobin < 10 g/dL and/or platelets < 100×109/L were identified in 23/25 (92%) patients. Most important non-hematologic signs of organ dysfunction included elevated alkaline phosphatase in 17/25 (68%) patients and splenomegaly (≥450 mL) or even marked splenomegaly (≥1200 mL) in 15/15 (100%) and 8/15 (53%) patients, respectively, with available spleen volumetry (n = 15). Mutations in KIT were identified in 22/25 (88%) patients (D816V, n = 18; D816Y, n = 2; D816H, n = 1; F522C, n = 1) with a median KIT D816V allele burden of 42% (range 20-98) in PB. Additional mutations in SRSF2, ASXL1 or RUNX1 (S/A/R gene panel, Jawhar et al., Leukemia 2016) were identified by NGS in 13/21 (62%) patients. Cytoreductive treatment in 22/25 patients included midostaurin (n = 21), cladribine (n = 9), and allogeneic stem cell transplantation (n = 1). Median observation from MCL diagnosis was 13 months (range, 0-86); 18/25 patients (72%) have died. Median overall survival (OS) was 17 months (range, 10-24) with a 2-year OS probability of 24% for all patients. OS of S/A/R mutated patients (n = 13) was significantly inferior as compared to S/A/R non-mutated patients (n = 8, P = 0.007, hazard ratio, HR: 5.8, 95% confidence interval: 1.3-26.1, median OS 13 months vs. not reached). Conclusion: Leukemic MCL and MCL without C-findings are rare, b) KIT D816V mutations were more frequently detected as previously reported, c) the prognostically relevant mutations in the S/A/R gene panel are present in approximately 60% of patients, and d) median OS is approximately 1.5 years with significantly inferior survival observed in S/A/R mutated patients. Disclosure: No conflict of interest disclosed.

Disclosure: Niklas Mueller: No conflict of interest disclosed. Peter Valent: Financing of Scientific Research: Novartis; Expert Testimony: Novartis

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

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V105

Incidence and prognostic impact of cytogenetic aberrations in systemic mastocytosis Naumann N.1, Jawhar M.1, Schwaab J.1, Metzgeroth G.1, Khaled N.1, Horny H.-P.2, Sotlar K.2, Valent P.3, Haferlach C.4, Göhring G.5, Schlegelberger B.5, Meggendorfer M.4, Cross N.C.P.6,7, Hofmann W.-K.1, Reiter A.1, Fabarius A.1 Department of Hematology and Oncology, University Medical Centre, Mannheim, Germany, 2Institute of Pathology, Ludwig-Maximilians-University, Munich, Germany, 3Department of Internal Medicine I, Division of Hematology and Ludwig Boltzmann Cluster Oncology, Medical University of Vienna, Vienna, Austria, 4MLL Munich Leukemia Laboratory, Munich, Germany, 5Institute of Human Genetics, Hannover Medical School, Hannover, Germany, 6Faculty of Medicine, University of Southampton, Southampton, United Kingdom, 7Wessex Regional Genetics Laboratory, Salisbury, United Kingdom 1

Introduction: In chronic myeloid neoplasms, the type, relative frequency and prognostic impact of cytogenetic aberrations are highly heterogeneous. With increasing knowledge on an adverse prognostic impact of specific gene mutations, contemporaneous presence and potential interaction of cytogenetic aberrations and mutations may become prognostically highly relevant. Methods: We analyzed clinical, cytogenetic and molecular characteristics of 109 (KIT D816V positive n = 102, 94%) patients (pts.) with indolent systemic mastocytosis (ISM, n = 27) and advanced SM (advSM, n = 82) with or without an associated hematologic neoplasms (AHN) [aggressive SM, n = 3; SM-AHN, n = 60; mast cell leukemia (MCL), n = 9; MCLAHN, n = 10)]. Results: In advSM, an aberrant karyotype (KT) was identified in 16/82 (18%) pts., which was complex (n = 7), associated with a bad prognosis (e.g. monosomy 7, n = 3) or associated with a good/unknown prognosis (e.g. +8 or del(5q), n = 6). In subsequent studies, KT anomalies were grouped and split into good KT (normal KT + good KT) and poor KT (complex KT + bad KT) based on e.g. MDS, AML. The median overall survival (OS) of pts. with advSM and poor KT (n = 10) was 4 months and significantly shorter than the OS of pts. with good KT (n = 72, 38 months) (HR 6.1, 95% confidence interval [2.6-14.2], p < 0.0001). Notably, progression into secondary AML or acute MCL was observed in 8/10 (80%) and 1/10 (10%) pts., median 4 months (range, 0.3-24) and 2 months after diagnosis of advSM and first detection of the cytogenetic aberration, respectively. Mutations in SRSF2/ASXL1/RUNX1 (S/A/Rpos.), which were recently shown to be associated with adverse OS in SM (Jawhar et al., Leukemia 2016), were identified in 5/10 (50%) pts. with poor KT and 31/72 (43%) pts. with good KT, respectively. Significant differences regarding OS were observed between good KT – S/A/Rneg. (n = 29) vs. good KT – S/A/ Rpos. (n = 31) vs. poor KT – S/A/Rneg. (n = 5) vs. poor KT – S/A/Rpos. (n = 5, Figure). Conclusion: In addition to clinical and morphological baseline staging, molecular and cytogenetic analyses should be routinely performed in pts. with advSM because they may indicate overt disease progression/transformation.

Oncol Res Treat 2016;39(suppl 3):1–104

Fig. 1. Disclosure: No conflict of interest disclosed. V106

Next-generation sequencing identifies MPL-TET2 mutated clones in a subset of patients with JAK2V617F-positive myelofibrosis Schulze S.1, Stengel R.2, Jäkel N.1, Wang S.-Y.1, Hubert K.3, Roskos M.2, Schneider M.2, Franke G.-N.1, Niederwieser D.1, Al-Ali H.K.1 Universitätsklinik Leipzig AöR, Hämatologie/ internistische Onkologie, Leipzig, Germany, 2Oncoscreen Synlab, Jena, Germany, 3Medizinische Fakultät/ Universität Leipzig, Hämatologie/ internistische Onkologie, Leipzig, Germany 1

The diagnostic workup via PCR or melting analysis is limited to analyzing exon 14 of JAK2, exon 10 of MPL, and exon 9 of CALR as the phenotype driver mutations (mut.) in JAK2, CALR, and MPL are considered to be mutually exclusive in patients (pts) with myelofibrosis (MF). We compared a next-generation sequencing (NGS)-based approach to standard PCR in detecting driver mut. Patients and methods: Genomic DNA was isolated from whole-blood samples of 129 pts with MF (68 males; median age 60y). JAK2, MPL, and CALR mut. analysis via PCR were performed as published. Then, NGS was performed to detect mut. in 23 genes known to be involved in myeloid malignancies. Results: Standard PCR detected JAK2V617F, MPLW515L, and mutated CALR in 81 (63%), 1 (0.8%), and 25 (19.4%) pts respectively. One patient (0.8%) harboured both mutated JAK2 and CALR. NGS identified further mut. in JAK2 [exon 24 at position R1063H (n = 6) and exon 20 at position R893T (n = 1)] in 7 JAK2V617F+ pts and in MPL at positions W515L + E335K, E259K, Y591D (n = 3), and Y591D (n = 2). None of the pts with JAK2R1063H or JAK2R893T mut. carried a TET2 or ASXL1 mut. JAK2TET2 double-mutant cells were present in 16 (12%) pts. Interestingly, 4 of 5 MPL mut. were found in the JAK2-TET2 mutated group and non in the CALR mutated or triple negative cohorts. Patients with the JAK2-MPLTET2 mut. were older, had a higher JAK2V617F allele burden, and larger spleens compared to the JAK2V617F+ “only” cohort (table).

Abstracts

CONTENTS AUTHOR INDEX

Tab. 1. Patients characteristics (n = 129)

Variable Median age (years) Median JAK2V617F allele burden, (%) Allele burden <50%, (%) Median palpable spleen, (cm) Median WBC x10e9/L Median peripheral blasts, (%) Hb < 100 g/L (%) DIPSS (Int-2/high-risk), (%)

JAK2V617F+ “only” cohort N = 65 60

JAK2-MPL-TET2 triple mutated cohort N = 4 69

9.3

16.5

42/15

25/25

Conclusions: With NGS, additional activating mut. such as MPLY591D (a gain of function mut. in exon 12 of MPL) could be detected. Our data provide evidence that driver mutations are not always mutually exclusive as was the case in 4% of the pts and imply that combination of mut. could influence the clinical phenotype. Analysis of larger cohorts will provide accurate estimates of the frequency of concomitant JAK2 and MPL mut. Serial NGS permits the study of the chronology of mut., which might influence the biology of the disease and/or response to therapy. Disclosure: Susann Schulze: No conflict of interest disclosed. Haifa Al-Ali: Financing of Scientific Research: yes; Expert Testimony: yes V107

Cell intrinsic induction of IP-10 in MPN Schnöder T.M.1, Eberhardt J.2, Nimmagadda S.C.2, Weinert S.3, Wolleschak D.2, Sammt A.2, Fahldieck C.2, Schönborn U.2, Fischer T.2, Heidel F.H.1,4 Universitätsklinikum Jena, Klinik für Innere Medizin II, Jena, Germany, 2Ottovon-Guericke Universität Magdeburg, Klinik für Hämatologie und Onkologie, Magdeburg, Germany, 3Otto-von-Guericke Universität Magdeburg, Klinik für Kardiologie und Angiologie, Magdeburg, Germany, 4Otto-von-Guericke Universität Magdeburg, Magdeburg, Germany 1

Introduction: Myeloproliferative neoplasm (MPN) is a group of diseases which include polycythemia vera (PV), essential thrombocytosis (ET) and primary myelofibrosis (PMF). In advanced phases, especially in post-ET and post-PV myelofibrosis, the clinical hallmark of the disease is a striking inflammatory syndrome. At a molecular level, PV, ET and PMF are characterized by an activating point mutation (V617F) of JAK2. Currently, the cellular and molecular basis of the inflammatory response syndrome is incompletely understood and therapeutic options for MPN patients ineligible for allogeneic stem cell transplantation are limited to symptomatic approaches. Here, we aimed to evaluate the role of JAK2V617F mutation for aberrant cytokine expression in MPN. Methods: IP-10 mRNA expression was analyzed in the peripheral blood of patients diagnosed with PV, ET or MF after informed consent using quantitative real-time PCR (qPCR). Moreover, BaF3 cells were stably transduced with EpoR and Jak2WT or Jak2V617F, respectively. Cytokine Array was performed to identify Jak2V617F-triggered inflammatory response. Validation of differentially expressed targets was performed using qPCR and immunoblotting. Results: IP-10 mRNA expression was significantly elevated in JAK2V617F+ MPN patients compared to healthy donors or JAK2V617FMPN patients. High V617F allele burden correlated with high IP-10 expression (p = 0.0003). To exclude paracrine/autocrine stimulation as a potential mechanism, we used BaF3 cells harboring the Jak2V617F mutation but lacking IP-10 receptor. IP-10 mRNA was also highly expressed in BaF3 Jak2V617F cells and pharmacological treatment with Ruxolitinib (RUX) abrogated IP-10 expression. NFκB signaling was in-

Abstracts

duced in Jak2V617F-positive cells as compared to Jak2WT controls and its activation was reduced after RUX treatment. Using a luciferase promoter assay, NFkB signaling was modulated by the presence or absence of Jak2V617F-mediated signaling, suggesting direct induction of IP-10 by NFkB signaling. Conclusions: Taken together, we could show that IP-10 expression correlates with JAK2V617F allele burden in MPN patients and is differentially up-regulated in the respective MPN entities. Our data provide first evidence for a link between oncogenic JAK2V617F signaling and cell intrinsic induction of chemokines/cytokines regulated through NFkB signaling. These data encourage further investigation to elucidate the intercellular pathomechanism of chronic inflammation in MPN. Disclosure: Tina Schnöder: No conflict of interest disclosed. Florian Heidel: Advisory Role: Advisory Board Novartis; Financing of Scientific Research: Vortragshonorar Novartis; Expert Testimony: Research Funding Novartis V108

Differential NK-suppressive capacity of the JAK2 inhibitor Pacritinib when compared to the JAK1/2 inhibitor Ruxolitinib Wolf D.1, Rudolph J.2, Brossart P.2, Schoenberg K.2 Universitätsklinikum Bonn, Bonn, Germany, 2Department of Internal Medicine III, Oncology, Hematology and Rheumatology, University Clinic Bonn (UKB), Bonn, Germany 1

Introduction: We previously demonstrated the NK cell suppressive potential of the JAK1/2 inhibitor ruxolitinib, which is also linked to infections (Schönberg et al, Cancer Res 2015). More specific JAK inhibitors may exert a differential immune-modulatory potential, leading to a less pronounced immunosuppression. The JAK2 inhibitor pacritinib is currently evaluated in phase III clinical trials. We here evaluated the NK cell suppressive potential of pacritinib as compared to ruxolitinib. Methods: NK cells from healthy volunteers were isolated by magnetic beads and purity was regularly controlled. NK cell function (killing, cytokine production), proliferation and receptor expression were evaluated by standard immunological techniques. The doses used were 0.1, 1 and 10 µM of both JAK inhibitors. The relevant in vivo achieved dose of ruxolitinib is 1-2µM, whereas the one of pacritinib is 0.1 µM. Results: In line with out previous results, ruxolitinib potently inhibits proliferation, activation, killing activity and cytokine production at doses of 1µM. When therapeutically achieved doses of pacritinib are compared to ruxolitinib, its NK suppressive potential is clearly lower, with only a slight but insignificant reduction of killing against K562 target cells and degranulation, whereas production of IFN-g is significantly reduced but clearly less potent when compared to ruxolitinib. More strikingly, 1 µM of ruxolitinib reduced proliferation by approximately 75%, whereas 0.1µM of pacritinib only marginally reduces NK cell proliferation. When comparing induction of apoptosis, ruxolitinib does not induce significant amounts of cell death up to doses of 10µM, whereas pacritinib at a concentration of 10µM almost completely eliminated NK cells. When NK cells are activated by activating NK receptor antibodies (i.e. NKp46mAb) instead of cytokine stimulation by IL-2, both compounds are ineffective with respect to NK cell suppression, which is again in line with our previous results showing that JAK-inhibitors predominantly inhibit cytokine-mediated NK cell activation. Conclusions: The more specific JAK2 Inhibitor pacritinib is less potent with respect to NK cell suppression when compared to the JAK1/2 inhibitor ruxolitinib. This may help to understand differential immune-modulatory effects of different types of JAK inhibitors. Disclosure: Dominik Wolf: Advisory Role: Bexalta, Novartis; Financing of Scientific Research: Bexalta, Novartis; Expert Testimony: Bexalta, Novartis Kathrin Schoenberg: No conflict of interest disclosed.

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Fortbildung

Immuntherapie in der Hämatologie und Onkologie V110

Checkpoint-inhibition in oncology Wolf D.1

prove specific T-cell functions. The CAR encoding gene is delivered into the T-cell using viral vectors and the final cell product is then re-transfused into the patient. CAR T-cell expansion upon antigen-recognition may result in severe cytokine release with relevant clinical symptoms like high fever and low blood pressure. Thus, CAR-T-cell therapy needs highly sophisticated logistics. This presentation will focus on the development of CAR T-cell therapy for malignant lymphomas.

Universitätsklinikum Bonn, Medizinische Klinik 3; Onkologie, Hämatologie, Immunonkologie und Rheumatologie, Bonn, Germany

Disclosure: No conflict of interest disclosed.

It was in 2011, when Hanahan and Weinberg renewed their initial “hallmark of cancer” concept by adding the ability of malignant tumors to escape from an efficient immune cell attack (termed cancer immune-evasion). It was already assumed for decades that tumors are able shape the immune system, thereby actively preventing their elimination. Visionaries such as Paul Ehrlich more than a century ago already envisioned that “magic (e.g. immunological) bullets” may overcome tolerance to cancer. Immune cell activation is fine-tuned by a number of receptor/ligand-pairs, such as immune-activating (CD28, GITR or CD137) and immune-inhibitory (PD1, CTLA-4, LAG3, TIM3) proteins. CTLA-4 and PD-1 on T and PD-L1 on cancer cells represent the most prominent checkpoint-molecules, since both targets can be succesfully blocked in a therapeutic setting by antibodies, leading to their approval in various solid metastatic diseases (melanoma, NSCLC and RCC). In addition, various proof-of-concept Phase 1/2 studies documented very encouraging response and survival rates in other tumors, (e.g. mismatch repair deficient colorectal cancer, Merkel cell carcinoma, ovarian cancer). Most strikingly, these compounds induce long-term survival in a subgroup of immune-susceptible patients. Clinically applicable response predictors are not available so far, even though in some tumors a greater overall survival can be seen in PD-L1 positive tumors. Moreover, it appears that “immunological visibility” based on the genetic instability appears to be associated with improved response rates to checkpoint-blocking agents. This explains superior response rates seen in smoking NSCLC patients carrying a higher mutational load leading to T cell activation based on the generation of mutated neo-antigens. The tolerability of checkpoint-antibodies is very good. In addition to infusion-related reactions, physicians particularly have to check for immune-related side-effects (e.g. pneumonitis, colitis, hypophysitis), as they may need a rapid therapeutic intervention with corticosteroids. In summary, immune-oncology (IO) represents a new therapy pillar in the treatment of solid tumors, which complements the classic treatment modalities of surgery, radiation and chemotherapy, as well as targeted drugs. Combination studies with classical therapeutics and different IO-agents fuel the hope that IO therapies will help to improve outcome of more cancer patients in the near future.

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Disclosure: Dominik Wolf: Financing of Scientific Research: BMS, MSD, Roche; Expert Testimony: BMS V112

Development of chimeric antigen receptor (CAR) T-cell immunotherapy Borchmann P.1 Uniklinik Köln, Klinik I für Innere Medizin, Köln, Germany

Chimeric antigen receptor (CAR) T-cell immunotherapy involves the adoptive transfer of patient-derived T-cells, which are genetically modified to express antigen-targeted receptors. The CAR is designed to recognize the tumor cell and induce a potent T-cell attack and therefore has extra- and intracellular domains. The extracellular domain usually consists of a tumor-antigen specific single-chain variable fragment coupled to a transmembrane linker. The intracellular signaling modules trigger the T-cell functions. One commonly used intracellular effector domain is the CD3ζ chain from the T-cell receptor; however, co-stimulatory signals are needed to ensure enhanced T-cell function and persistence. Co-stimulatory domains include CD28, 4-1BB, OX40, ICOS. There are many different options to combine different intracellular effector domains in order to im-

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Mammakarzinom: (Neo)adjuvante Therapie V114

AGO recommendations for the diagnosis and treatment of patients with early breast cancer: update 2016 Janni W.1 Universität Ulm, Ulm, Germany

For the last 15 years, the Breast Committee of the Arbeitsgemeinschaft Gynäkologische Onkologie (German Gynecological Oncology Group, AGO) has been preparing and updating evidence-based recommendations for the diagnosis and treatment of patients with early and metastatic breast cancer. The AGO Breast Committee consists of gynecological oncologists specialized in breast cancer and interdisciplinary members specialized in pathology, radiological diagnostics, medical oncology, and radiation oncology. The lecture will give an overview of the 2016 update, which has been performed according to a documented rule-fixed algorithm, by thoroughly reviewing and scoring chapter by chapter the recent publications for their scientific validity (Oxford level of evidence (LoE), www.cebm.net) and clinical relevance (AGO grades of recommendation (GR); table 1). I will present a summary of the 2016 update; the full version of the updated slide set is available online as a PDF file in both English and German. Disclosure: No conflict of interest disclosed.

Fortbildung ZNS-Tumoren V117

The 2016 WHO classification of brain tumors: What’s new? Riemenschneider M.J.1 Universitätsklinikum Regensburg, Abteilung für Neuropathologie, Regensburg, Germany 1

Since its first launch in 1979 the WHO classification of tumors of the (central) nervous system has been continuously developed. In 1993 immunohistochemistry had been added and in 2000 the classification was extended by the description of molecular alterations though diagnoses still remained based on histology and immunohistochemistry alone. The ratio for brain tumor therapies in the meantime, however, has radically changed. Clinicians on a regular basis ask for a number of well-established molecular markers and patients often are primarily treated according to molecular factors rather than to histological diagnoses. It is in this context, that the new 2016 (revised 4th) edition of the classification describes a major paradigmatic change: For the first time, molecular parameters will be defining for certain diagnoses and an integrated histological and molecular diagnosis becomes state-of-the-art for some brain tumor entities. This makes perfect sense for molecular parameters such as combined allelic deletions on chromosome arms 1p/19q that are closely associated with defined histological appearances. Consequently, oligoastrocytoma will be a dying entity that will prospectively only exist in a histological context with molecular diagnostics not yet performed. Also, the IDH mutational status will be part of an integrated diagnosis, stigmatizing a

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diffusely infiltrating IDH-wildtype astrocytoma with non-glioblastoma histology as a weird tumor with divergent histological and molecular features. It is a major strength of the new classification that such tumors that clinically might behave irregularly are now readily identified and can be treated accordingly. They can also be omitted from clinical studies making groups more homogeneous and results more meaningful. In this context, also new entities like the genetically defined medulloblastomas might open up options for more individualized therapies both inside and outside of clinical studies. While most academically-based brain tumor centers with specialized neuropathology departments should be able to promptly implement the new procedures in their diagnostic work-up, the new classification poses considerable challenges to smaller local medical centers. Though an NOS category is possible in settings where molecular tests cannot be performed, collaborative clinical networks are more than before recommended to secure best possible diagnoses and up-to-date treatment options for all patients affected. Disclosure: No conflict of interest disclosed.

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Kontroverse Indikationen zur Stammzelltransplantation V119

Allogeneic stem cell transplantation as primary treatment of multiple myeloma Mielke S.1 Universitätsklinikum Würzburg, Medizinische Klinik und Poliklinik II, Zentrum für allogene Blutstammzelltransplantation, Würzburg, Germany 1

Standard treatment for fit patients with multiple myeloma (MM) includes induction therapy followed by autologous stem cell transplantation (SCT) providing substantial treatment-free intervals for the majority of patients. Although there is promising evidence of an allogeneic immune system’s principal ability to provide long term immune-surveillance or even cure to patients suffering from MM, there is ongoing discussion whether allogeneic SCT represents a standard of care treatment option for fit and younger patients. With low rates of non-relapse mortality (NRM) in place, which is largely a consequence of the introduction of reduced-intensity conditioning regimens and improved supportive care options, constantly high rates of relapse and possibly impaired long-term quality of life (QoL) aspects drive today’s controversial discussions particularly in light of several new treatment options surfacing. Nevertheless the genetic instability and the clonal evolution associated with the progression of MM leave allogeneic SCT more than ever as a valuable treatment option to achieve long-term disease control. This however, does require an optimal selection of patients most likely benefitting from this approach. Here, the scenario of primary allogeneic stem cell transplantation for patients with high risk disease upfront after induction therapy or closely following autologous SCT as a tandem approach will be discussed in the light of results for allogeneic SCT as a salvage treatment for patients relapsing after autologous SCT. With regard to dramatically increasing health care costs associated with the approval of several new drugs in the field of MM allogeneic SCT may represent indeed a very reasonable treatment option also from an economical point of view particularly when being capable of offering cure to patients. Certainly, these questions have to be answered in prospective clinical trials in the context of new drugs used for induction treatment before and possibly for consolidation after allogeneic SCT.

V122

Allogeneic hematopoietic stem cell transplantation (HSCT) with an unrelated donor as first-line therapy in patients with severe aplastic anemia (SAA) Tichelli A.1, Passweg J.1 Universitätsspital Basel, Hämatologie, Basel, Switzerland

Allogeneic HSCT is standard first-line treatment for patients with SAA younger than 40 years with an eligible matched sibling donor (MSD). Patients not eligible for HSCT should receive immunosuppressive therapy (IST). Candidates for a matched unrelated donor (MUD) are patients having failed at least one course of IST. The outcome of unrelated HSCT has greatly improved, as the consequence of better supportive care, improved conditioning regimens and superior donor selection by high resolution HLA-typing. Therefore, the paradigm on algorithm of first-line therapy in SAA is questioned. So far, most data on MUD HSCT are of retrospective nature and based on patients who failed first-line IST. Compared to MSD, transplants from MUD lead to more acute and chronic GVHD, but survival is not inferior. The negative predictors for survival are the same in MSD and MUD HSCT: the use of peripheral blood, longer interval from diagnosis to HSCT, older age at transplantation. Excellent results are obtained with a 8/8 or 10/10 MUD, but significantly inferior with < 8/8 MUD transplants. In a recent French cohort study, three risk factors (age >30 years; interval longer >12months; 9/10 mismatch), have been integrated into a risk score. The 4-year survival was 74% for patients with low-risk and 49% for patients with high-risk score. In a retrospective study, 29 children receiving upfront MUD transplant were compared to 87 MSD HSCT and 58 IST. The 2-year overall survival was >90% for all groups, but event-free survival was significantly lower for IST controls. Concern about upfront MUD transplantation in SAA is the delay in processing from diagnosis to HSCT. The median time interval from diagnosis to MUD HSCT was 4.5 months, which is barely longer than the time needed for MSD HSCT. Histocompatibility laboratories are able to estimate the probability to identify a compatible unrelated donor. About 30% of patients have a high probability to find fast a MUD. In conclusion, time for revision of the current treatment algorithm for first-line therapy of SAA patients has come. An upfront 10/10 MUD HSCT should be considered as first-line treatment for children and young adults, who lack a MSD, provided there is a high chance for fast donor availability. If a suitable MUD is available to donate bone marrow within 3-4 months from diagnosis, upfront MUD transplantation seems reasonable. The risk score proposed by the French group could help in patient selection for an upfront MUD HSCT. Disclosure: No conflict of interest disclosed.

Disclosure: Stephan Mielke: Financing of Scientific Research: Celgene, Cellex; Other Financial Relationships: Celgene

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V124

Lebensqualitäts-Assessment

Electronic collection of quality of life data – one option to suit them all?

V123

Holzner B.1, Sztankay M.2, Gamper E.-M.1, Rumpold G.3, Wintner L.M.1

Assessing quality of life in clinical trials (CTs) - what should be paid attention to? Engelhardt M.1, Ihorst G.2, Waldschmidt J.1, Keller A.1, Deschler B.3, Dold S.1, Zober A.1, Messner C.1, Möller M.1, Wünsch A.4, Wirsching M.5, Antes G.6, Vach W.7, Duyster J.1, Wäsch R.1 University of Freiburg Medical Center, Hematology & Oncology + Comprehensive Cancer Center Freiburg (CCCF), Freiburg, Germany, 2Clinical Trials Center, University of Freiburg, Freiburg, Germany, 3Department of Internal Medicine II, CCC Mainfranken, University Hospital Würzburg, Würzburg, Germany, 4Technische Universität München, München, Germany, 5Department of Psychosomatic Medicine and Psychotherapy, University Medical Center, Freiburg, Germany, 6Cochrane Center University of Freiburg, Freiburg, Germany, 7 Center for Medical Biometry and Medical Informatics, University of Freiburg, Freiburg, Germany 1

Introduction: Growing budget pains caused by new products and concerns about novel drugs´ financial toxicity have fueled an acrimonious debate over explosive costs. Health technology assessments (HTA) have been included in CTs, providing additional information on Quality of life (QoL) and health care resource utilization. Albeit these QoL assessments are desirable, if not mandatory, doubts persist about their relevance to clinical practice and which tools should be used. Methods: We assessed all relevant literature via search functions “QoL”, “CTs”, “oncology”, “State-of-the-art”, obtaining ~30.000 references, limited to “last 5 yrs, humans, English, core journals and cancer”. Moreover, of >130 cancer CTs in our hem&onc clinic, all were screened for quality measures; and compared to results obtained within functional comorbidity assessments (CMAs), initially used in MDS/AML and currently in multiple myeloma (MM) patients (pts). Results: Pt-reported outcomes (PROs) allow to include pt perspectives into CTs. Many PRO survey instruments are however lengthy and few facilitate care management. The analysis of available QoL tools demonstrate, that most cancer CTs use the EORTC QLQ-C30 or shorter SF-12 with most consistently sensitive QoL scales being appetite loss, fatigue, pain, role, social and physical dysfunctions. Core principles for the development of health care quality measures should address 1. pt-centered outcomes, 2. support robust scientific evidence linked to improved health outcomes, 3. include anticipated benefits, 4. balance in time and resources required to acquire the data. CMA instruments in older pts with MDS/AML have been implemented in wise decision-making; our brief prospective CMA in MM pts includes the IADL, ADL, Timed Up and Go Test, malnutrition, pain, fitness, SF12 and depression. This CMA is performed to assess pts´ fitness, tolerability of multiagent treatment and to predict OS. Conclusion: QoL indicators remain relevant, their perfection being under extensive debate. Today, treatment decisions need to be driven by efficacy, QoL being affected, SAE risks, pt preference and costs/affordability. Most predictive CMA tools should be determined and consistently included in future CTs. Scientific advice sessions with regulators, HTA experts and drug developers seem vital, discussing premarketing CT designs openly to support effective and safe new treatments. Disclosure: Monika Engelhardt: Advisory Role: Janssen, Celgene, Novartis, MSD, Amgen; Financing of Scientific Research: Janssen, Celgene, Novartis, MSD, Amgen; Expert Testimony: Janssen, Celgene, Novartis, MSD, Amgen Ralph Wäsch: Advisory Role: Celgene, Novartis, MSD, Amgen; Financing of Scientific Research: Celgene, Novartis, MSD, Amgen

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Medical University of Innsbruck, Department of Psychiatry, Psychotherapy and Psychosomatics, Innsbruck, Austria, 2University of Innsbruck, Institute of Sport Science, Innsbruck, Austria, 3Medical University of Innsbruck, Department of Medical Psychology, Innsbruck, Austria 1

Patient-reported outcomes (PRO) like quality of life add the patient’s perspective to traditional clinical outcomes and facilitate patient-centred care. The feasibility of electronic PRO collection (ePRO) grows with decreasing costs for technical devices, comprehensive availability of internet services and rising acceptance of computer-based measures and internet-enabled devices, even in the elderly. In Austria, about 40% of persons older than 65 years regularly access the internet and nearly as many cancer patients stated having internet access and its occasional use. Overall, ePRO is well accepted and only a small proportion of patients state a preference for paper-pencil measures (ranging from 10-18%). ePRO offers several benefits, as it simplifies data collection procedures, improves data quality, eases and accelerates data calculation, presentation and storage and saves human resources. Available software differs considerably concerning features, applicability and adaptability to the particular needs of users respecting the individual clinical workflow. Useful functionalities include PRO monitoring in daily clinical routine, study monitoring, clinical data bases, and patient portals. One major benefit of routinely collected ePRO data is its versatility, enabling research with real word data, health economy and quality assurance analyses. The Computer-based Health Evaluation System (CHES) is an ePRO software used for various clinical conditions (e.g. oncology, orthopaedics, neurology, psychiatry/psychosomatics, dementia) in multiple institutions worldwide (e.g. medical universities, rehabilitation centres, county hospitals in AUT, CH, GER, CAN, the UK). Within the longstanding collaboration with the EORTC Quality of Life Group and the EBMT, CHES has shown to be a reliable and user-friendly software for daily clinical routine and research. Literature shows that ePRO in oncology is feasible both technically and contentwise. The success of ePRO applications depends on reliable software as well as a proper implementation strategy considering several aspects (e.g. clear purpose and expectations, throughout inclusion of stakeholders, adaption of software, training and process evaluation). Systems with integrated patient portals offer an additional benefit to patients, providing disease and treatment-related information, education and self-management advice tailored to PRO results to empower active involvement into their own care. Disclosure: Bernhard Holzner: Honoraria: Bernhard Holzner and Gerhard Rumpold are owner of the intellectual property rights of the software CHES. Lisa M. Wintner: No conflict of interest disclosed. V126

Assessment of quality of life in patients with aplastic anemia and/or PNH Panse J.1 Universtiätsklinikum Aachen, Onkologie, Hämatologie und Stammzelltransplantation, Aachen, Germany 1

Acquired Aplastic Anemia (AA) and Paroxysmal Nocturnal Hemoglobinuria (PNH) are so-called ultra-rare diseases. While much is known about patho-physiology and treatment of these interrelated diseases, less is known about pts. psycho-social issues. Quality of life (QoL) evaluation tools used in studies for AA and PNH are rather unspecific and were designed for cancer patients (pts) (e.g. the EORTC QLQ-C30). Given the complexity of AA and PNH, the variation in symptoms and treatment approaches, the young age of the pts, and the fact that marrow failure syndromes are not classified as malignant diseases, it is likely that cancer-spe-

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cific questionnaires are inappropriate to adequately assess the QoL and illness intrusiveness in these pts. Hence, we initiated the development and validation of an AA/PNH-specific QoL-instrument (QLQ-AA/PNH). The generation of a QLQ-AA/PNH was performed according to EORTC QoL group guidelines: after identification of QoL issues by literature review, a focus group of pts and physicians were interviewed (phase I). After screening of documented interviews, QoL issues were generated and reworded in a preliminary questionnaire (phase II). In phase III the questionnaire with generated items was tested in a representative patient group and psychometrically validated. In addition, patients were asked to complete a questionnaire regarding their supportive care needs, (e. g. information, support by medical staff, psychosocial counseling, patient support groups etc.) and potential iatrogenic problems (e. g. delay in diagnosis, appreciation of QoL problems by health care professionals). 102 patients in more than 25 German and Swiss cities were interviewed. In phase I, interviews of 19 pts and 8 physicians specialized in AA/PNH-treatment resulted in 649 QoL issues; these were condensed to 175, and graded according to their importance by 30 pts and 14 physicians (phase II). 97 issues were rated important. 12 EORTC QLQ-C30 items were not rated important, while several new QoL aspects were brought up. Modifications led to two questionnaires with 77 items regarding general QoL-aspects and 20 items regarding medical care. Pre-testing of the questionnaire including 97 items was done with 52 pts, followed by psychometric validation. The final QLQ-AA/PNH includes 54 questions. It has been translated into English, Italian and French and is currently validated through phase III studies for AA pts and by use of the PNH registry for PNH pts. Disclosure: No conflict of interest disclosed.

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Eosinophilie, Mastozytose V127

Differential diagnosis and therapy of hypereosinophilia Metzgeroth G.1 III. Medizinische Klinik, Hämatologie und internistische Onkologie, Universitätsmedizin Mannheim, Mannheim, Germany 1

Sustained eosinophilia (>0.5×109/l) is observed in a wide range of reactive/non-clonal and neoplastic/clonal disorders. The clinical presentation may be complicated by a life-threatening organ damage, e.g. of heart, lung and/or nervous system. In the majority of cases, eosinophilia is reactive, e.g. due to an autoimmune disorder or hypereosinophilic syndrome (HES), through overproduction of eosinophilopoietic cytokines. In the presence of distinct molecular aberrations, the underlying entities are either classified as ‘myeloid/lymphoid neoplasms associated with eosinophilia and rearrangements of PDGFRA, PDGFRB, FGFR1 or JAK2 (MLNeo)’ or ‘chronic eosinophilic leukemia, not otherwise specified (CELNOS)’, other cases as eosinophilia-associated myeloproliferative neoplasm (MPN-eo). The FIP1L1-PDGFRA fusion gene, which is only identified by FISH or RT-PCR, is the most frequent molecular aberration. Cytogenetic analysis from a bone marrow aspirate is mandatory for the detection of rearrangements of 5q31-33 (PDGFRB), 8p11 (FGFR1) or 9p24 (JAK2) and subsequent identification of fusion partners by FISH/PCR. FIP1L1-PDGFRA negative patients with normal karyotype should be screened for KIT D816V and JAK2 V617F leading to diagnosis of systemic mastocytosis (SM-eo) and MPN-eo, respectively. All those patients are candidates for treatment with tyrosine kinase inhibitors, however, sustained responses are not seen in all entities. Patients with PDGFRA/-B fusion genes achieve durable remissions on imatinib, irrespective of the disease stage (chronic or blast phase). Resistance, e.g. due to T674I or D842V point mutations in PDGFRA, is extremely rare. Responses may also be observed in patients with JAK2 fusion genes on ruxolitinib and to a lesser extent in patients with FGFR1 fusion genes on ponatinib, however, resistance and/or progression are common. Because of an aggressive clinical course, eligible patients should be offered an early allogeneic stem cell transplantation.

Abstracts

In CEL-NOS and MPN-eo, hydroxyurea and possibly also imatinib or ruxolitinib remain treatment options, SM-eo should be treated with midostaurin and/or cladribine. In reactive eosinophilia with organ damage, the disease is usually diagnosed as autoimmune disorder or HES. It is most important to find a good balance between response and potential adverse effects of corticosteroids. Careful attention should therefore be paid to the need of steroid-sparing drugs such as MTX, azathioprin or cyclophosphamide. Disclosure: No conflict of interest disclosed. V130

Treatment options in systemic mastocytosis Reiter A.1, Jawhar M.1, Metzgeroth G.1, Schwaab J.1 Universitätsmedizin Mannheim, Hämatologie und Onkologie, Mannheim, Germany 1

Systemic mastocytosis (SM) is a clonal hematologic neoplasm characterized by accumulation of neoplastic mast cells (MC) in various tissues, e.g. bone marrow (BM), visceral organs and skin. The diagnosis is based upon the presence of aggregates of spindle shaped mast cells with abnormal immunophenotype (CD117+/CD25+), elevated serum tryptase levels and presence of the KIT D816V mutation in 80-90% of patients. Indolent SM (ISM) shows little or no evidence of organ damage and a normal life expectancy. Treatment is based on the avoidance of triggers and symptomatic treatment with H1- and H2-inhibitors, cromoglycin acid and steroids. Advanced SM (advSM) comprises several subtypes that are characterized by the overlapping presence of an associated hematologic neoplasm (SMAHN, predominantly myelodysplastic/myeloproliferative or myeloproliferative neoplasms, SM-MDS/MPN, SM-MPN), C-findings (e.g. cytopenia, hypoalbuminemia, malabsorption, ascites, weight loss) indicating organ damage (aggressive SM, ASM) or >20% mast cells in BM smears (mast cell leukemia, MCL). More than 80% of advSM patients have one or more (>60%) additional mutations in genes such as TET2, SRSF2, ASXL1, RUNX1, CBL, JAK2, RAS and others. Mutations in SRSF2, ASXL1 and/ or RUNX1 have a strong negative impact on phenotype, clinical course and prognosis. Organ damage and clinical symptoms frequently require rapid initiation of cytoreductive therapy. For many years, only IFN-alpha and cladribine have been available achieving variable degrees of clinical remissions, which are rarely complete and durable. Currently, the main focus lies on KIT-inhibitors, e.g. midostaurin, and allogeneic stem cell transplantation (ASCT). In a phase-II-study, midostaurin induced significant responses (C-findings, BM infiltration, serum tryptase levels, clinical symptoms) in the majority of patients, independently of the subtype, KIT mutation status or prior therapy. A retrospective study of 57 patients revealed that ASCT was associated with responses and complete remissions in 40 (70%) and 16 (28%) of patients, respectively. Overall survival at 3 years was 57% for all patients, 74% for SM-AHN, 43% for ASM and 17% for MCL, respectively. Future studies should gain a better insight into the impact of the complex molecular profile on treatment outcome and they should evaluate combined treatment strategies including conventional cytoreductive therapy, KIT inhibitors and ASCT. Disclosure: Andreas Reiter: Advisory Role: Novartis Pharma; Financing of Scientific Research: Novartis Pharma Juliana Schwaab: Financing of Scientific Research: Novartis Pharma

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Freier Vortrag

Tab. 1.

Nichtkleinzelliges Lungenkarzinom V131

Summary of EGFR-Biomarker data from the randomized, controlled phase 3 trial SQUIRE on adding Necitumumab to first-line Gemcitabine-Cisplatin (GC) for squamous Non-Small Cell Lung Cancer (sqNSCLC) Schumann C.1, Reck M.2, Thomas M.3, Mezger J.4, Socinski M.A.5, Hozak R.6, Mi G.6, Depenbrock H.7, Krause T.7, Hirsch F.R.8, Thatcher N.9 Klinikum Kempten, Pneumologie, Thoraxonkologie, Schlaf- und Beatmungsmedizin, Kempten, Germany, 2LungenClinic Großhansdorf, Department of Thoracic Oncology, Airway Research Center North (ARCN), German Center for Lung Research (DZL), Großhansdorf, Germany, 3Internistische Onkologie der Thoraxtumoren, Thoraxlinik im Universitätsklinikum Heidelberg, Translational Lung Research Center Heidelberg (TLRC-H), Heidelberg, Germany, 4 St. VincentiusKliniken, Medizinische Klinik II, Karlsruhe, Germany, 5University of Pittsburgh, Department of Hematology/Oncology, Pittsburgh, United States, 6 Eli Lilly and Company, Indianapolis, United States, 7Lilly Deutschland GmbH, Medical – Oncology, Bad Homburg, Germany, 8University of Colorado Cancer Center, Aurora, United States, 9The Christie Hospital, Manchester, United Kingdom 1

Introduction: In SQUIRE (N = 1093), the addition of the IgG1 EGFR-antibody necitumumab to first-line GC in advanced sqNSCLC significantly improved overall survival (OS); the safety profile was acceptable (Thatcher et al., Lancet Oncol 2015;16:763-774). SQUIRE included mandatory tissue collection from archived tumor. Here, we summarize analyses exploring the relationship of EGFR protein expression and EGFR gene copy number with efficacy outcomes (OS, PFS). Methods: Tissue samples were centrally assessed for EGFR protein expression by immunohistochemistry (IHC, Dako EGFR PharmDx kit) and for EGFR gene copy number by fluorescence in situ hybridization (FISH). OS and PFS were evaluated by Kaplan-Meier analysis and Cox proportional hazard models, separately for patients with EGFR-protein expressing (EGFR>0) and non-expressing (EGFR = 0) tumors (stratified, pre-specified analysis), and for patients with and without increased EGFR gene copy number (Hirsch et al., J Clin Oncol 2008;26:3351-3357; exploratory, unstratified interaction analysis). Results: Tissue samples were evaluable for EGFR protein expression in 982 of the 1093 randomized patients (89.8%). The majority of these patients (935/982, 95.2%) had EGFR-expressing tumors, and this subgroup benefitted from the addition of necitumumab similar to the overall population (Table). Benefit was not apparent in the small subgroup of patients with non-EGFR expressing tumors. Samples for EGFR gene copy analysis were available for 557 patients (51.0%). Of these, 208 (37.3%) had an increased EGFR gene copy number (FISH positive tumor). Patients with increased EGFR gene copy number showed more favorable hazard ratios for both OS and PFS (0.70 and 0.71, respectively; vs. 1.02 and 1.04 for non-increased copy number, but with a non-significant treatment-by-marker interaction (Table). Conclusions: Patients with EGFR expressing tumors benefitted from the addition of necitumumab to first-line GC treatment. EGFR gene copy number gain showed a trend for a more favorable HR, but the interaction was not statistically significant. Further investigations to better understand the roles of these biomarkers are ongoing.

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Disclosure: Christian Schumann: No conflict of interest disclosed. Nick Thatcher: Financing of Scientific Research: Eli Lilly and Company V132

Distribution of histological subtypes of lung cancer in Germany (1999-2012) Hermann S.1, Friedrich S.1, Arndt V.1 Deutsches Krebsforschungszentrum (DKFZ), Epidemiologisches Krebsregister Baden-Württemberg, Heidelberg, Germany 1

Introduction: Earlier publications from other countries have shown that the proportion of adenocarcinoma lung cancer has increased while squamous cell carcinoma has decreased over the last decades presumably due to changes in cigarette design, composition and smoking habits. Our aim is to evaluate if this shift in histological subgroups has also taken place in Germany. Methods: The study is based on all primary lung cancer cases (ICD-10 C34) diagnosed in 1999–2012 and reported by six German epidemiological cancer registries (Bremen, Hamburg, Mecklenburg-Vorpommern, Münster, Sachsen, Saarland) after exclusion of death certificate only cases. Histology of lung cancer cases was classified into: squamous cell carcinoma (SCC), adenocarcinoma (ADC), small cell carcinoma (SCLC), large cell carcinoma (LCC), other, and unspecific morphology. Overall, sex-, and age-specific trends in the distribution histological subtypes was assessed. Results: A total of 87.558 lung cancer cases (males: 63.109, females: 24.449), diagnosed 1999–2012, were included. The overall distribution according to histological subtype was ADC 26.9%, SCC 24.2%, unspecific histology 22.7%, LCC 7.2%, SCLC 3.6%, and other 15.4%. After exclusion of cases with unspecific histology, the proportion of ADC continuously increased significantly between 1999 and 2012 in men (25.6%→38.2%) and in women (37.0%→50.5%) whereas the proportion of SCC significantly decreased from 39.5% to 32.4% in men and from 22.1% to 16.2% in women. This pattern was consistent over all age-groups, least pronounced for patients ≥75 years of age and most pronounced for age-group 65-74 years of age. In absolute numbers, ADC increased between 1999 and 2012 from 1158 to 2114 cases (+ 134% in women, + 60% in men) whereas the

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number of all other lung cancer cases with specified histology addition increased by 36% in women and decreased by 10.5% in men. Conclusion: As in other countries, both the proportion as well as the absolute number of adenocarcinoma lung cancer has increased in Germany between 1999 and 2012. Increases in ADC are considered to reflect a wider use of filter-tripped and low-tar cigarettes. Acknowledgement: The here used data originates from six cancer registries (Bremen, Hamburg, Mecklenburg-Vorpommern, Münster, Sachsen, Saarland) and was provided via the German Centre for Cancer Registry Data. Our thanks goes to each individual cancer registry for the contribution of the data. Disclosure: No conflict of interest disclosed. V133

P53 non-disruptive mutation is a negative predictive factor in EGFR M+ NSCLC treated with TKI Griesinger F.1, Netchaeva M.1, Lüers A.1, Prenzel R.2, Scriba D.3, Willborn K.C.4, Stropiep U.5, Hallas C.6, Tiemann M.6, Falk M.6, Neemann N.7, Heukamp L.7, Roeper J.1 Pius-Hospital Oldenburg, University of Oldenburg, Universitätsklinik Innere Medizin-Onkologie, Oldenburg, Germany, 2Pius-Hospital Oldenburg, Department Pneumology, Oldenburg, Germany, 3Pius-Hospital Oldenburg, Department Thoracic Surgery, Oldenburg, Germany, 4Pius-Hospital Oldenburg, Department Radiotherapy, Oldenburg, Germany, 5Pius-Hospital Oldenburg, Oldenburg, Germany, 6Hematopathology Hamburg, Hamburg, Germany, 7New Oncology, Köln, Germany 1

Introduction: P53 mutations are common in lung cancer, and have also been described in EGFR M+ patients. The impact of p53 mutations in EGFR M+ patients is controversial, especially if classified as disruptive and non-disruptive according to their functional effect on the p53 protein as proposed by Poeta and colleagues. The aim of the study was therefore to systematically analyze EGFR and p53 mutations within a cohort of patients with lung cancer stage IV (UICC 7), to correlate alterations with clinical characteristics and to investigate a potential impact of p53 mutations on treatment outcome. Methods: 409 patients from a single center diagnosed with lung cancer stage IV were studied for the presence of EGFR as well as inactivating p53 mutations. Methods for the detection of EGFR mutations included Sanger Sequencing and hybridization based COBAS testing, hybrid cage next generation sequencing. P53 mutations were detected by Sanger Sequencing and either MiSeq or hybrid cage NGS. Clinical characteristics including smoking status were available for more than 95% of the patients. Results: 409 consecutive patients at the lung cancer center of the Pius-Hospital Oldenburg were studied. The overall EGFR M+ rate was 18% (73/409) in all patients, 73% (53/73) showing common mutations of exon 19 or 21. In 21/73 (29%) patients’ p53 analysis was not successful. P53 disruptive mutations were demonstrated in 25% (13/52) of successfully tested patients, and p53 non-disruptive mutation occurred in 31% (16/52) whereas p53 WT configuration was found in 44% (23/52). Median OS was 28 months in p53 disruptive mutation and 42 month in p53 WT compared to 23 months in p53 non-disruptive mutation (p < 0.018). PFS on 1st line TKI therapy was 14 months in p53 disruptive mutation, 18 months in p53 WT and 7 months in p53 non-disruptive mutation (p < 0.001). Similar results were shown in the EGFR common mutation but not in the uncommon mutation subgroup. The ORR was 49.2% in patients with a disruptive p53 mutation/p53 WT constellation compared to 14.3% in patients with a non-disruptive p53 mutation and 17.5% in patients with an unknown p53 status. Conclusion: Significant differences in PFS and OS in EGFR M+ patients were observed depending on p53 mutation status. P53 mutational status is only predictive when disruptive and non-disruptive p53 mutations are differentiated. P53 should be tested prospectively in EGFR M+ patients as management of patients on 1st line TKI may be different.

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A comprehensive analysis of potentially targetable genetic aberrations and clinical findings in 821 patients with squamous-cell NSCLC – a comparison of NGM and TCGA LUSC data Koleczko S.1, Schäpers C.1, Scheffler M.1, Ihle M.2, Kostenko A.3, Michels S.1, Fischer R.1, Nogova L.1, Serke M.4, Kaminsky B.5, Benedikter J.6, Brümmendorf T.H.7, Ficker J.H.8, Lorenz J.9, Schulte C.10, Schulze-Olden S.11, Brandes V.1, Abdulla D.1, Ueckeroth F.2, Thurat M.1, Merkelbach-Bruse S.2, Büttner R.2, Wolf J.1 LCGC, Universitätsklinikum Köln, Köln, Germany, 2Institut für Pathologie, Universitätsklinikum Köln, Köln, Germany, 3Klinik I für Innere Medizin – NGM, Universitätsklinikum Köln, Köln, Germany, 4Lungenklinik Hemer, Hemer, Germany, 5Krankenhaus Bethanien, Solingen, Germany, 6Städtisches Klinikum München, München, Germany, 7RTWH Aachen, Aachen, Germany, 8Klinikum Nürnberg, Nürnberg, Germany, 9Klinikum Lüdenscheid, Lüdenscheid, Germany, 10 Gemeinschaftspraxis für Hämatologie und Onkologie, Dortmund, Germany, 11 Florence-Nightingale-Krankenhaus der Kaiserswerther Diakonie, Düsseldorf, Germany 1

Introduction: In contrast to the improvements which have been made in the treatment of adenocarcinoma NSCLC, squamous-cell carcinoma NSCLC (SCC) remains a therapeutic challenge. While there are recent advantages with immunotherapy approaches, targeted therapy still lacks of evidence regarding the frequency of driver aberrations in advanced SCC. We set out this study in order to characterize a large-scale set of patients with SCC genetically and clinically and compared the findings to the early-stage The Cancer Genome Atlas (TCGA) LUSC cohort. Methods: Tumor biopsies of 821 patients were analyzed within the Network Genomic Medicine (NGM) lung cancer using next-generation parallel sequencing (NGS). The panel used consisted of 102 amplicons and 14 genes: KRAS, PIK3CA, BRAF, EGFR, ERBB2, NRAS, DDR2, TP53, ALK, CTNNB1, MET, AKT1, PTEN and MAP2K1. In subsets of patients, fluorescence in-situ hybridization (FISH) was performed for amplification detection of FGFR1 and MET. We queried the TCGA dataset with respect to the panel used and compared the findings. For the NGM patients, therapy and outcome were also collected. Results: Beside expected frequencies of TP53, DDR2, PTEN and PIK3CA mutations, we detected EGFR mutations in 3.2% and BRAF mutations in 1.8%. Unlike the TCGA dataset, where the frequencies were 2.8% and 3.9%, respectively, the detected mutations in the NGM cohort consisted of activating mutations (i. e., EGFR del19 and L858R, and BRAF V600E). FISH data revealed a presence of MET amplification in 14.2% and of FGFR1 amplification in 20.0%. The association and correlation of these aberrations with clinical findings and prognosis as well as with PD-L1 expression status and mutational load is still being analyzed. HER2 amplification, which occurred in 2.2% of the TCGA cohort, will be analyzed in a subset of patients. Conclusions: Our data suggest that the presence of a potential targetable aberration might occur in up to 40% of SCC all-comers. Further analyses are warranted in order to characterize SCC patients according to their biomarker profiles for potential treatment recommendations. Disclosure: Sophia Koleczko: No conflict of interest disclosed. Jürgen Wolf: Advisory Role: ASTRAZENECA, Boehringer Ingelheim, Bristol-Myers Squibb, Clovis, MSD, Novartis, Pfizer, Roche; Financing of Scientific Research: siehe oben; Expert Testimony: MSD, Novartis, Pfizer, Roche; Other Financial Relationships: siehe oben

Disclosure: No conflict of interest disclosed.

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

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CheckMate 017 and 057 studies of nivolumab vs docetaxel in patients with advanced NSCLC: 2-year-update and exploratory cytokine profile analyses Eberhardt W.E.E.1, Borghaei H.2, Brahmer J.R.3, Horn L.4, Ready N.5, Steins M.6, Felip E.7, Paz-Ares L.G.8, Arrieta O.9, Barlesi F.10, Antonia S.J.11, Fayette J.12, Rizvi N.A.13, Crinò L.14, Reck M.15, Hellmann M.13, Desai K.16, Li A.16, Healey D.16, Spigel D.R.17 University Hospital – University Duisburg-Essen, West German Tumor Center, Essen, Germany, 2Fox Chase Cancer Center, Philadelphia, United States, 3The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, United States, 4Vanderbilt University Medical Center, Nashville, United States, 5 Duke University Medical Center, Durham, United States, 6ThoraxklinikHeidelberg gGmbh, Heidelberg, Germany, 7Vall d’Hebron University Hospital, Vall d’Hebron Institute of Oncology, Medical Oncology Department, Barcelona, Spain, 8Virgen del Rocio University Hospital, Seville, Spain, 9Instituto Nacional de Cancerologia – INCAN, Mexico City, Mexico, 10Aix-Marseille University, Assistance Publique Hôpitaux de Marseille, Hopital Nord, Marseille, France, 11H. Lee Moffitt Cancer Center, Tampa, United States, 12Centre Léon Bérard, Lyon, France, 13Memorial Sloan Kettering Cancer Center, New York, United States, 14 Ospedale di Perugia, Perugia, Italy, 15Lungen Clinic Grosshansdorf, Airway Research Center North (ARCN), Grosshansdorf, Germany, 16Bristol-Myers Squibb, Princeton, United States, 17Sarah Cannon Research Institute, Nashville, United States 1

Background: Nivolumab, a fully human IgG4 programmed death-1 immune checkpoint inhibitor, is approved in the US and EU for patients with previously treated advanced/metastatic NSCLC, based on results of 2 phase 3 trials: CheckMate 017 (NCT01642004) in squamous (SQ) NSCLC and CheckMate 057 (NCT01673867) in non-SQ (NSQ) NSCLC. Methods: Patients were randomized 1:1 to receive nivolumab 3 mg/kg Q2W or docetaxel 75 mg/m2 Q3W until progression or discontinuation. OS was the primary objective in both studies. Multivariate analyses of baseline serum cytokines (exploratory) were performed separately in patients with SQ and NSQ NSCLC. Cytoscores derived from evaluable patients in three studies (CheckMate 017, 057, and 063 [a single-arm phase 2 study of nivolumab in SQ NSCLC] were calculated to quantify the effect of identified cytokine sets on 18-month OS. Cytoscores were categorized as high or low based on median cutoffs. Results: CheckMate 017: median OS was 9.2 vs 6.0 months with nivolumab vs docetaxel (18month OS: 28% vs 13%; HR: 0.62 [0.48, 0.81]; P = 0.0004). CheckMate 057: median OS was 12.2 vs 9.4 months with nivolumab vs docetaxel (18-month OS: 39% vs 23%; HR: 0.72 [0.60, 0.88]; P = 0.0009). Patients with NSQ NSCLC that expressed PD-L1 achieved a greater magnitude of benefit with nivolumab; in patients with SQ NSCLC, PD-L1 expression was neither prognostic nor predictive of benefit. In both trials, treatment-related AEs were less frequent with nivolumab than with docetaxel. Preliminary results showed an association of high vs low SQ- and NSQ-cytoscores with OS (SQ NSCLC: nivolumab, HR = 0.48, P < 0.0001; docetaxel, HR = 0.39, P < 0.0001; NSQ NSCLC: nivolumab, HR = 0.52, P = 0.0001; docetaxel, HR = 0.60, P = 0.0001; figure). Conclusion: Nivolumab demonstrated improved OS and a favorable safety profile compared with docetaxel in patients with SQ and NSQ NSCLC. Two-year OS and safety results will be presented. Levels of baseline serum cytokines (details to be presented) correlated with OS benefit in patients with advanced SQ and NSQ NSCLC. Prospective validation of these preliminary results is needed.

Oncol Res Treat 2016;39(suppl 3):1–104

Fig. 1. Borghaei DGHO. Disclosure: Wilfried Ernst Erich Eberhardt: Advisory Role: Consulting/advisory role: BMS, Eli Lilly, Astra Zeneca, Pfizer, Roche, Novartis, Boehringer-Ingelheim, Merck, Hexal, Bayer, Astellas, Daichi Sankyo; Financing of Scientific Research: BMS, Eli Lilly, Astra Zeneca, Pfizer, Roche, Novartis, Boehringer-Ingelheim, Merck, Hexal, Bayer, Astellas; Expert Testimony: Eli Lilly; Other Financial Relationships: Speakers Bureau: Roche, Pfizer, Novartis, Merck, Boehringer-Ingelheim David Spigel: Advisory Role: Consulting/advisory role: Novartis/Genentech; Other Financial Relationships: Travel, accommodations, expenses: Novartis, Genentech, Pfizer; Speakers Bureau: Novartis (uncompensated)

Fortbildung

Interprofessionelle Sitzung: Therapiebegrenzung (für Ärzte und Pflegekräfte) V138

The EPAL-project (Ethics policy for advanced care planning and limiting treatment) Winkler E.1, Mehlis K.1, Jaeger E.2, Laryionava K.1, Hiddemann W.2, Heussner P.2 Universität Heidelberg, Nationales Centrum für Tumorerkrankungen, Heidelberg, Germany, 2Klinikum der Ludwig-Maximilians Universität, Medizinische Klinik und Poliklinik III, München, Germany 1

Introduction: Decisions to limit treatment (DLT) are important in order to protect the patient from burdensome treatment at the end of life but constitute one of the most ethically challenging situations in oncology practice. Thus, they are often accompanied by clinical and psychological conflicts for patients and the medical team. The aim of the EPAL-project is to investigate relevant factors influencing End-of-life decision-making and to develop an ethics policy for these decisions. The policy intends to establish a framework for responding to patients’ preferences in an ethically responsible way and supporting the concept of advance care planning. Methods: This prospective quantitative study recruited 50 cancer patients from 5/2014-6/2015 in the Dept. of Hematology/Oncology at the University Hospital of Munich with treatment limitations. The patients and their respective physician and nurse completed a set of validated instruments on moral distress, patient’s information needs, involvement in DLT and perceived quality of communication. Results: Nurses state to have moral distress in 71% (n = 30) of the cases. The average level of stress lies under the cut-off point of 5 (mean/SD: 1.8/1.7) and especially occurs in situations, when DLT is surrounded by a challenging communication within the medical team (p = 0.047). Nurs-

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es do not know patients’ preferences on treatment limitation in 77% and only 61% (n = 30) of the patients feel sufficiently involved in DLT. Those who show a higher degree of information seeking behavior are more often involved (p = 0.037). Patients perceiving a good communication feel significantly better informed about their diagnosis (p = 0.003), treatment options (p = 0.019) and progress of the disease (p = 0.000) and are more satisfied with the decision-making process (p = 0.010) compared to those perceiving a bad interaction. Conclusion: Given the fact that good interaction within the medical team and with the patient is strongly related to the nurses’ level of stress and the patients’ level of information, participation and satisfaction concerning DLT, there is a need to ensure good communication and respect for patient autonomy in these crucial situations. Our policy focuses on these aspects and aims to define conditions and standards for early discussions about treatment limitation, shared decision-making and a respectful communication between physicians, nurses, patients and relatives. A post-implementation study will evaluate the policy’s impact. Disclosure: No conflict of interest disclosed. V140

Referral to palliative care without prognostic awareness – how to deal with diaappointment Schuler U.S.1, Freitag J.1 Universitätsklinikum Carl Gustav Carus, PalliativCentrum und Medizinische Klinik I, Dresden, Germany 1

Prognostic understanding, prognostic awareness of patients is frequently lacking even in advanced stages either due to insufficient communication or due to denial. Even the trial of early integration of palliative care (Temel et al. 2011, JCO 29:2319) reported a rate of ~20% of patients regarding their cancer as curable at 12-24 weeks in the intervention group. Lack of prognostic awareness comes in two variants: (1) The complete lack of information about missing curative options and (2) the acceptance of incurability combined with unrealistic expectations about survival duration. PCUs have to deal with both variants, as important decisions arise. Even at admission, it is sometimes necessary to confront patients with issues like do-not-resuscitate orders (DNR). Other issues (e.g. unnecessary medication, artificial nutrition etc.) are left for clarification in the ensuing days. Even if no longer indicated for physical reasons, these may be continued at least for some time for psychosocial reasons, until cessation becomes acceptable for all parties involved. It is of utmost importance to include (wherever permitted by the patient) close relatives in the decision making. Relinquishing artificial nutrition sometimes leads over-ambitious relatives to almost force-feed their relatives. Especially nutrition is an issue, which frequently involves nursing staff in further communication. But for all goals of care, it is important to keep the multidisciplinary team informed and communication with the patient and family transparent and consistent. Representative examples will be given. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Chronische lymphatische Leukämie – Therapie V141

Ibrutinib (I) plus bendamustine and rituximab (BR) in previously treated chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL): a 2-year follow-up of the HELIOS study Cramer P.1, Fraser G.2, Chanan-Khan A.3, Demirkan F.4, Santucci Silva R.5, Pylypenko H.6, Grosicki S.7, Janssens A.8, Pristupa A.9, Mayer J.10, Dilhuydy M.-S.11, Loscertales J.12, Goy A.13, Avigdor A.14, Rule S.15, Phelps C.16, Mahler M.16, Salman M.16, Howes A.17, Balasubramanian S.16, Hallek M.1 Uniklinik Köln, Klinik I für Innere Medizin, Köln, Germany, 2McMaster University, Juravinski Cancer Centre, Hamilton, Canada, 3Mayo Clinic, Division of Hematology and Oncology, Jacksonville, United States, 4Dokuz Eylul University, Division of Hematology, Izmir, Turkey, 5IEP SÃO LUCAS, Hemomed Oncologia e Hematologia, São Paulo, Brazil, 6Cherkassy Regional Oncological Center, Division of Hematology, Cherkassy, Ukraine, 7Community of Hospitals, Division of Hematology, Chorzow, Poland, 8Universitaire Ziekenhuizen Leuven, Leuven, Belgium, 9Regional Clinical Hospital, Ryazan, Russian Federation, 10University Hospital Brno, Division of Internal Medicine, Hematology and Oncology, Jihlavska, Czech Republic, 11Hopital Haut Leveque, Bordeaux, France, 12Hospital Universitario La Princesa, IIS-IP, Division of Hematology, Madrid, Spain, 13John Theurer Cancer Center, Hackensack University Medical Center, Hackensack, United States, 14The Chaim Sheba Medical Center, Institute of Hematology, Division of Hematology and Bone-Marrow Transplantation, Tel Hashomer, Israel, 15Derriford Hospital, Division of Hematology, Plymouth, United Kingdom, 16 Janssen Research & Development, Raritan, United States, 17Janssen Research & Development, High Wycombe, United Kingdom 1

Introduction: The phase 3 HELIOS study evaluated I + BR vs placebo (P) + BR in patients (pts) with previously treated CLL/SLL. At first analysis (median follow-up: 17.0 months) progression-free survival (PFS) was significantly improved for I + BR vs P + BR (HR [95% CI]: 0.203 [0.150.28], p < 0.0001). Prior studies have shown deepening responses with continued ibrutinib treatment; long-term follow-up assessing durability and depth of response is important. Methods: 578 pts received BR (≤ 6 cycles) and were randomized 1:1 to I (420 mg daily) or P (n = 289/arm). Pts with del17p (≥ 20% of cells) were excluded. Primary end point was PFS. Key secondary end points: overall survival (OS), overall response rate (ORR), and rate of minimum residual disease negative (MRD -ve) response. Results: Median follow up is 25.4 months. I + BR continues to show improvement in PFS vs P + BR (investigator [INV]-assessed median: not reached [NR] vs 14.2 months; HR [95% CI]: 0.199 [0.15, 0.26], p < 0.0001; 2-yr rate: 74.8% vs 20.9%). Median OS is still NR in either arm (HR [95% CI]: 0.670 [0.44, 1.02], p = 0.0587; 2-yr rate: 86.2% vs 81.5%); 142 pts (49.1%) in the P + BR arm with confirmed PD have crossed over to receive I, as permitted by protocol. The updated INV-assessed ORR is 87.2% for I + BR vs 66.1% for P + BR (p < 0.0001); updated rates of CR/CRi are 33.9% vs 7.2% (rates at first analysis: 21.4% vs 5.9%). Rates of MRD -ve response for the intent-to-treat population are 18.0% (52/289) for I + BR and 4.8% (14/289) for P + BR (p < 0.0001) (rates at first analysis: 12.8% vs 4.8%). Median PFS2 is unreached in both arms, but PFS2 is significantly longer for pts assigned to I + BR vs P + BR, despite crossover (HR [95% CI]: 0.622 [0.42, 0.92], p = 0.0162). Safety is consistent with first analysis. Conclusions: I + BR continues to demonstrate superiority vs P + BR with significantly longer PFS and higher ORR. Responses continue to deepen with continuous ibrutinib therapy with rates of CR/CRi and MRD -ve response increasing over time. At every MRD level (< 0.01% or ≥10% CLL+), I + BR showed a more sustained PFS than P + BR. These 2-year follow-up data confirm the important role of ibrutinib in pts with previously treated CLL. Trial Registration: NCT01611090; EudraCT No. 2012-000600-15; UTN No. U1111-1135-3745

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

CONTENTS AUTHOR INDEX

Disclosure: Paula Cramer: Advisory Role: Janssen, Hoffman-LaRoche; Financing of Scientific Research: Speakers Bureau: Janssen, Hoffman-LaRoche; Expert Testimony: Gilead, GSK, Janssen, Hoffman-LaRoche; Other Financial Relationships: Travel grants: Astellas, Gilead, Janssen, Hoffman-LaRoche, MundiPharma Michael Hallek: Advisory Role: Abbvie; Financing of Scientific Research: Abbvie; Expert Testimony: Abbvie V142

In CLL a complex karyotype is an adverse prognostic parameter that is associated with shorter survival independent of TP53 alteration and IGHV mutation status Haferlach C.1, Jeromin S.1, Kern W.1, Haferlach T.1 MLL Münchner Leukämielabor, München, Germany

Introduction: First studies demonstrated a prognostic impact of chromosome banding analysis (CBA) in addition to FISH, TP53 mutation and IGHV mutation status (Haferlach et al. GCC 2010, Rigolin et al. Blood 2012, Thompson et al. Cancer 2015). The aim of this study was to evaluate the impact of a complex karyotype in untreated CLL patients. Patients and methods: 1046 CLL cases were evaluated at diagnosis or prior to first therapy by CBA and FISH with probes for 17p13 (TP53), 13q14 (D13S25, D13S319, DLEU), 11q22 (ATM), the centromere of chromosome 12 and IGH-CCND1. Further, the TP53 and IGHV mutation status was determined in all patients. Median age was 67 years (range: 30-89 years). Results: Patients were categorized according to the number of chromosome abnormalities (CA) detected per case by CBA. None, 1, 2, 3, 4 and ≥5 abnormalities were observed in 228 (21.8%), 449 (42.9%), 208 (19.9%), 72 (6.9%), 36 (3.4%), and 53 (5.1%) cases, respectively. In 46 (4.4%) cases a TP53/17p deletion was identified by FISH and in 76 (7.3%) TP53 mutations by sequencing. 39 patients carried both a TP53 deletion and a TP53 mutation. A significant association was observed between the number of CA and TP53 alterations: 0-2 CA: 5.5%, 3 CA: 11.1%, 4 CA: 13.9%, and ≥5 CA 39.6% (p < 0.001). The number of CA was also associated with an unmutated IGHV status (p < 0.001). OS at 5 years differed significantly according to the number of CA (0-2 CA: 84.8%, 3 CA: 83.7%, 4 CA: 77.7%, ≥5 CA: 52.6%, p < 0.001). Three different definitions for complex karyotypes (CK) were evaluated: CK3: ≥3 CA, CK4: ≥4 CA, and CK5: ≥5 CA. In patients without a TP53 alteration overall survival (OS) at 5 years was significantly lower in cases harboring a CK than in those without (CK3: 80.3% vs 86.4%, p = 0.03; CK4: 73.6% vs 86.4%, p = 0.004; CK5: 66.4% vs 86.3%, p = 0.001). Also in patients with TP53 alterations a CK was associated with a negative impact on OS (OS at 5 years: CK3: 41.0% vs 59.6%, p = 0.01; CK4: 35.7% vs 59.4%, p = 0.01; CK5: 31.3% vs 59.1%, p = 0.002). In multivariate Cox regression analysis an independent negative impact on OS was identified for TP53 deletion (relative risk (RR): 3.1, p = 0.001), TP53 mutation (RR: 1.7, p = 0.05), number of chromosome abnormalities (RR: 1.1 per CA, p = 0.006) and unmutated IGHV status (RR: 2.0, p < 0.001). Conclusion: The number of CA as determined by CBA is a negative prognostic parameter in CLL which is independent of the presence of TP53 alterations and the IGHV mutation status. Disclosure: No conflict of interest disclosed. V143

The Pan-PIM kinase inhibitor LGB321 targets apoptotic pathways and microenvironmental interactions in CLL Decker S.1, Kissel S.1, Aumann K.2, Zenz T.3, Zirlik K.1, Claus R.1, Duyster J.1, Dierks C.1 Uniklinik Freiburg/Innere Medizin/Hämatologie, Onkologie und Stammzelltransplantation, Sektion Molekulare Hämatologie, Freiburg, Germany, 2Uniklinik Freiburg, Pathologie, Freiburg, Germany, 3NCT Heidelberg, Heidelberg, Germany 1

In recent years, the emergence of kinase inhibitors like Ibrutinib has drastically altered treatment strategies and improved outcomes in CLL patients, but lack of cure and resistance to therapy still remain serious

Oncol Res Treat 2016;39(suppl 3):1–104

problems. PIM kinases are involved in various important disease mechanisms in CLL, with PIM1 regulating CXCR4 surface expression impacting its interaction with the microenvironment, and PIM2/3 affecting the apoptotic machinery by regulating BAD. The Pan-PIM kinase inhibitor LGB321 (Novartis) targets all 3 PIM kinases and therefore affects both, CLL apoposis and its interaction with the microenvironment. In the study presented here, we investigated the effect of LGB321 on CLL in vitro and in vivo. LGB321 was highly effective in inducing apoptosis in primary CLL cells, independent of risk factors like 17p deletions or the mutation status. Apoptosis induction correlated with reduced pBAD and BAD levels. Furthermore, LGB321 was also effective in the presence of protective stromal cells and could completely overcome the stroma protective effects. Mechanistically, we found that LGB321 treatment blocked the CXCR4/ CXCL12 axis by dephosphorylating the CXCR4 receptor on Ser339, by reducing total CXCR4 protein levels and by blocking the externalization of the CXCR4 receptor. Concordantly, PIM inhibition blocked CXCR4 functions like migration towards a CXCL12 gradient (P < .0001), and reduced homing of LGB321-pretreated primary CLL cells towards the bone marrow (P = 0.0001) of NOG mice in vivo. In vivo experiments comfirmed the efficacy of LGB321 in 4 different CLL xenograft studies. Transplantation of primary human CLL cells into NOG mice and treatment with LGB321 for 2 weeks strongly reduced WBC counts, spleen size and spleen infiltration with human CLL cells (P = 0.0295) in all four CLL cases, and blocked BAD as well as CXCR4 phosphorylation also in vivo. Our results demonstrate, that the Pan-PIM kinase inhibitor LGB321 might be an effective treatment option for CLL patients by impairing PIM2/3 mediated CLL-cell survival, and by blocking the PIM1/CXCR4-mediated interaction of CLL cells with their protective microenvironment in vitro and in vivo. Future clinical trials should be performed to validate its efficacy in human CLL. Disclosure: No conflict of interest disclosed. V144

Checkpoint inhibition reverses immunometabolic dysfunctions of monocytes in chronic lymphocytic leukemia Mougiakakos D.1, Qorraj M.1, Bruns H.1, Böttcher M.1, Saul D.1, Jitschin R.1, Mackensen A.1 Medizinische Klinik 5, Universitätsklinikum Erlangen-Nürnberg, Erlangen, Germany 1

Introduction: CLL patients display immune defects at early disease stages. Increasing evidence suggests that immune alterations negatively impact disease course. The intrinsic tumor immune surveillance is severely attenuated. Monocytes are key components of anti-tumor immunity and important mediators of effects triggered by therapeutic antibodies. The myeloid compartment of CLL patients displays substantial alterations. However, the underlying mechanisms remain largely elusive. It is well accepted that the immune cells’ metabolism strongly determines differentiation and (anti-tumor) function. Current reports suggest that a “metabolic interplay” between cancer and T-cells contributes to tumor immune escape. As yet, nothing has been reported on immunometabolic dysfunctions of monocytes (in CLL). Similar to activated effector T-cells monocytes require an increased glycolytic flux for functioning efficiently. Methods: CLL-monocytes were divided into classical (CD14++CD16-), intermediate (CD14++CD16+), and non-classical (CD14-CD16++) subsets. We assessed the metabolic phenotype of these subsets using gene-expression, FACS-based, and functional analyses. The impact of metabolic interference on the monocytes’ phagocytic function was tested using E. coli and CLL-cells opsonized with a therapeutic anti-CD20 antibody. The role of PD-L1/PD-1 crosstalk in mediating immunometabolic alterations was investigated in co-cultures of CLL-cells and monocytes. Results: CLL-monocytes exhibit several changes of their metabolic phenotype: glucose uptake, glucose transporters, and expression of key glycolytic molecules are significantly reduced. Increasing the glycolytic rate enhances the monocytes’ efficacy in terms of eliminating primary CLLcells. PD-1/PD-L1 interaction is involved in the CLL-mediated T-cell sup-

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pression (=immune checkpoint). In a recent study, PD-1/PD-L1 pathway activation was shown to elicit metabolic reprogramming in T-cells by inhibiting glycolysis. We observed similar effects when triggering PD-1 signaling on monocytes. As anticipated, disrupting the PD-1/PD-L1 axis reversed the metabolic and phagocytic dysfunctions of (CLL-)monocytes. Conclusions: Our data suggests an immunometabolic crosstalk between CLL-cells and monocytes that hampers the monocytes’ function. It provides a strong rationale for exploiting PD-1/PD-L1 checkpoint blockade (e.g. in combination with anti-CD20 antibodies) to restore metabolic together with antitumor activity of monocytes. Disclosure: No conflict of interest disclosed. V145

HLA ligandome analysis of primary chronic lymphocytic leukemia (CLL) cells under lenalidomide treatment approves lenalidomide as a suitable adjuvant for T-cell based immunotherapy Nelde A.1, Kowalewski D.J.1, Backert L.2, Schuster H.1, Kanz L.3, Salih H.R.3,4, Rammensee H.-G.1,5, Stevanovic S.1,5, Stickel J.S.3 Universität Tübingen, Abteilung für Immunologie, Tübingen, Germany, Universität Tübingen, Center für Bioinformatik und Abteilung für angewandte Computerwissenschaften, Tübingen, Germany, 3Universitätsklinik Tübingen, Hämatologie und Onkologie, Tübingen, Germany, 4Clinical Cooperation Unit Translational Immunology, German Cancer Consortium (DKTK), DKFZ partner site Tübingen, Tübingen, Germany, 5German Cancer Consortium (DKTK), DKFZ partner site Tübingen, Tübingen, Germany 1 2

Several studies have proven the positive immunomodulatory effect of lenalidomide on T-cell responses in CLL, in particular regarding the repair of the immune synapse between CLL and T cells. Therefore, lenalidomide seems to be a promising adjuvant for T-cell based immunotherapy approaches. We recently conducted a study, which characterized the antigenic landscape of CLL by mass spectrometric analysis of naturally presented HLA ligands and identified a panel of CLL-specific T-cell epitopes (Kowalewski/PNAS 2015). For a rational combination of T-cell based immunotherapy with lenalidomide, it is of great importance to characterize the effects of this immunomodulatory drug on the antigenic landscape of the target cells. Here we present a mass spectrometry-based study, which longitudinally maps the HLA ligandome of primary CLL cells under in vitro lenalidomide treatment. We quantified HLA surface expression on CLL cells (n = 4) at t0, t24h and t48h after incubation with lenalidomide. With regard to HLA class I expression, no impact of lenalidomide treatment was observed (fold-change 0.92-1.02, t48h), whereas a slight increase of HLA class II molecules after treatment was detectable (fold-change 1.25-1.43, t48h) with absolute molecule counts ranging from 40,000-125,000 class I and 30,000-200,000 class II molecules/cell. Implementing label-free quantification data, we then quantitatively assessed HLA class I ligand presentation during lenalidomide treatment. We observed a higher plasticity of the HLA ligandome over time compared to treatment with lenalidomide. 2.5 ± 3.0% of HLA class I ligands showed significant modulation (fold change ≥4, p ≤ 0.01) after 24h of mock treatment, whereas only 0.9 ± 1.2% of the ligands were modulated through lenalidomide treatment. At t48h similar proportions of modulation were observed with 4.0 ± 1.7% of HLA ligands significantly altered in their abundance over time, while lenalidomide treatment only resulted in 0.9 ± 1.2% modulated ligands. Out of the >6300 different HLA class I ligands we could identify on primary CLL cells (n = 3), we were able to detect 39 CLL-associated epitopes described in our previous study. Importantly, these antigens showed robust presentation under lenalidomide therapy. Taken together our study shows that lenalidomide has no relevant influence on and rather seems to stabilize the HLA ligandome of primary CLL cells. Therefore, lenalidomide appears to be an ideal adjuvant for T-cell based immunotherapy in CLL patients.

V146

Clinical response to ibrutinib is accompanied by normalization of the T-cell environment in CLL-related autoimmune cytopenia Schliffke S.1, Akyüz N.1, Ford C.1, Mährle T.1, Thenhausen T.2, Krohn-Grimberghe A.2, Knop S.3, Bokemeyer C.1, Binder M.1 Universitäsklinikum Hamburg-Eppendorf, Hamburg, Germany, 2LYTIQ GmbH, Paderborn, Germany, 3Universitätsklinikum Würzburg, Würzburg, Germany 1

B-cell receptor-targeted therapy with ibrutinib is of increasing importance for the therapeutic management of chronic lymphocytic leukemia (CLL). However, limited data is available regarding the therapeutic potential of ibrutinib in the treatment of autoimmune cytopenias (AIC), which frequently affect patients with CLL. The pathophysiological mechanisms driving AIC in CLL are insufficiently understood, but the majority of evidence points towards a polyclonal B- and T-cell-mediated response, rather than direct involvement of the malignant clone. Beyond targeting of malignant B-cells, ibrutinib has a broader immune modulating profile through inhibition of Bruton’s tyrosine kinase (BTK) and the interleukin-2-inducible T-cell kinase (ITK) in a variety of immune cells and – in line with this – ibrutinib is currently under investigation for autoimmune disorders. This was the rationale behind our decision to initiate treatment with ibrutinib in three cases of severe CLL-related AIC. In addition to close clinical monitoring, immune profiling from these patients with multicolor-flow cytometry and next generation sequencing (NGS) of the B- and T-cell repertoire was performed. Pre-treatment analysis by flow cytometry suggested exceptionally high levels of effector memory CD8+ T-cells and activated CD8+ T-cells compared to a cohort of 100 CLL patients without AIC. In parallel with rapid clinical improvement, effector memory and activated CD8+ T-cell counts normalized after the initiation of ibrutinib. Circulating B-cells, however, were almost exclusively of CLL phenotype pre- and post-treatment. Deep sequencing of the immunoglobulin heavy chain and the T-cell receptor beta chain locus revealed circulating T-cell immune environments that were heavily biased towards hyperexpanded and – most likely – autoimmunity-related T-cell clones. After initiation of ibrutinib, these T-cell immune environments normalized significantly with a loss of the hyperexpanded clones. In contrast, the B-cell immune environment remained essentially unchanged with dominance of the malignant CLL clone throughout the treatment. We believe that this observation provides additional evidence that CLL-related AIC is driven by abnormal T-cell environments that can be effectively targeted with ibrutinib and that this supports a clinical trial of early treatment with ibrutinib in CLL-related AIC. Furthermore our data point to response kinetics that vary throughout different ibrutinib target structures. Disclosure: No conflict of interest disclosed.

Fortbildung

Intensivmedizin und Hämatologie/Onkologie V150

Hemophagocytic Lymphohistiozytosis (HLH): A Mimic of Sepsis La Rosée P.1 Schwarzwald-Baar Klinikum, Villingen-Schwenningen, Germany

Disclosure: No conflict of interest disclosed.

Hemophagocytic Lymphohistiocytosis (HLH) is a hyperinflammatory syndrome, which depending on disease severity can mistakenly be diagnosed as SIRS, Sepsis or multiorgan dysfunctional syndrome (MODYS) on an intensive care unit (ICU). It is a cytokine storm syndrome affecting in particular immunosuppressed patients, and patients with malignant or autoimmune diseases. While HLH is better known in pediatrics due to well characterized hereditary defects of the immune synapse, HLH in adult patients only recently comes into focus of hematologists and ICU physicians. Diagnostic criteria developed for pediatric trials (HLH-2004)

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

CONTENTS AUTHOR INDEX

are cytopenia, organomegly, fever, elevated Ferritin and soluble IL2-receptor, hemophagocytosis, loss of NK-cell function and lowered fibrinogen or/and elevated triglyzerides. When 5/8 criteria are rated positive, the probability of HLH is high. It is recommended to include S-Ferritin into the lab panel for ICU admission. Early recognition and rapid suppression of overt inflammation are critical for prognosis. Hence, suppression of the cytokine storm by polyvalent immunoglobulins, corticosteroids and etoposide in selected patients, and maintenance with cyclosporine A or tacrolimus under certain circumstances are therapeutic strategies with evidence from case series. Recently, targeted agents like Jak1/2-inhibitors, Interferon-gamma antibodies or IL6-antibodies have been used in early phase clinical trials or case reports. It is important to note that definitive treatment of the underlying trigger disease is the only way to permanently control HLH. Close cooperation of ICU-physicians, hematologists, rheumatologists, infectious disease specialists and diagnostic institutions is pivotal for successful treatment. Management of HLH in adults is discussed based on clinical cases derived from the German HLH registry www.hlh-registry.org.

Conclusively, recent data question the benefits of NIMV in hematologic patients with hypoxic ARF. Furthermore, NIMV failure is common and associated with mortality. If NIMV is used in such patients, meticulous screening for risk factors associated with NIMV failure and early evaluation of intubation in case of their occurrence seems advisable. HFNO seems to be of benefit in non-immunocompromised patients with hypoxic ARF. Confirmation of these data in cancer patients is needed.

Disclosure: Paul La Rosée: Advisory Role: Novartis Pharma; Financing of scientific Research: Novartis Pharma

Treatment free remission in CML patients after first line nilotinib therapy

V151

Eigendorff E.1, Saussele S.2, Gattermann N.3, Le Coutre P.4, Illmer T.5, Brümmendorf T.H.6, Giles F.7, Hochhaus A.1

Non-invasive mechanical ventilation for prevention of invasive mechanical ventilation? Schellongowski P.1, Wohlfarth P.1, Sperr W.R.2, Knöbl P.2, Buchtele N.1, Gelbenegger G.1, Rabitsch W.3, Staudinger T.1, AG für HämatoOnkologische Intensivmedizin der ÖGIAIN; AK für Intensivmedizin in der Hämatologie und Onkologie der DGHO; Intensive Care in Hematologic and Oncologic Patients (iCHOP) Universitätsklinik für Innere Medizin I, Medizinische Universität Wien, Intensivstation 13i2, Wien, Austria, 2Universitätsklinik für Innere Medizin I, Medizinische Universität Wien, Abteilung für Hämatologie und Hämostaseologie, Wien, Austria, 3Universitätsklinik für Innere Medizin I, Medizinische Universität Wien, Knochenmarktransplantation, Wien, Austria 1

The acute respiratory failure (ARF) is the most frequent reason for intensive care unit (ICU) admissions in cancer patients. It is of strong prognostic impact, especially, if invasive mechanical ventilation becomes necessary. Non-invasive mechanical ventilation (NIMV) is established for the prevention of intubation in case of cardiac pulmonary edema and obstructive pulmonary disease. However, solid data on patients with hypoxic ARF due to pneumonia or sepsis – both most relevant in cancer patients – are lacking. A small historic trial showed reduced intubation and mortality rates in immunosuppressed patients with hypoxic ARF (n = 52) receiving pressure support NIMV when compared to O2 insufflation. However, as mortality rates of control group patients were very high, these results may not be relevant nowadays anymore (Hilbert, NEJM 2001). Observational studies revealed inhomogeneous results regarding the association of NIMV, intubation and mortality in hematologic patients, and identified risk factors for NIMV failure (=intubation): severity if illness, ARDS, high respiratory rate during NIMV, delay between admission and NIMV, vasopressors or renal replacement therapy, NIMV dependency > 72h, nonrapidly reversible or unknown causes for ARF, or presence of airway involvement by the malignancy. A meta-analysis showed successful NIMV to be associated with increased survival. However, NIMV failure was common (61%) and associated with increased risk of death (Amado-Rodríguez, Crit Care Med 2016). A recent trial in immunosuppressed, mainly hematologic patients (n = 374) with hypoxic ARF compared NIMV with O2 insufflation and found no differences of intubation or survival rates (Lemiale, JAMA 2015). These results may have been influenced by the unequal use of high-flow nasal oxygen (HFNO) between the groups, as HFNO was shown to reduce intubation and 90-day mortality rates in non-immunocompromised patients with hypoxic ARF when compared to NIMV or O2 insufflation (Frat, NEJM 2015).

Oncol Res Treat 2016;39(suppl 3):1–104

Disclosure: Peter Schellongowski: Financing of Scientific Research: Fisher & Paykel Thomas Staudinger: No conflict of interest disclosed.

Freier Vortrag

Chronische myeloische Leukämie – Erstlinien- und Absetzstudien V153

Klinik für Innere Medizin II, Universitätsklinikum, Jena, Germany, 2III. Medizinische Klinik, Universitätsmedizin, Mannheim, Germany, 3Klinik für Hämatologie, Onkologie und Klinische Immunologie, Universitätsklinikum, Düsseldorf, Germany, 4Medizinische Klinik m.S. Hämatologie und Onkologie, Charité, Berlin, Germany, 5BAG Freiberg-Richter, Jacobasch, Illmer, Wolf, Dresden, Germany, 6Klinik für Onkologie, Hämatologie und Stammzelltransplantation, Universitätsklinikum, Aachen, Germany, 7Division of Hematology/Oncology Northwestern University, Chicago, United States 1

Introduction: Tyrosine kinase inhibitors (TKI) are the standard of care for patients (pts) with chronic myeloid leukemia in chronic phase (CPCML). Although TKI therapy may result in a stable deep molecular response (DMR) in a significant proportion of pts, CML management guidelines recommend indefinite continuation of TKI therapy. According to current opinion, pts are eligible for TFR studies after >3 y of TKI therapy including at least one y of stable DMR (MR4.5, BCR-ABL ≤0.0032% on the International Scale [IS]) and known BCR-ABL transcript type. Within ENESTnd (NCT00471497), these crucial, but not exclusive prerequisites were met in 37.9 vs 21.6% of pts after nilotinib (NIL) 2*300 mg/d vs imatinib 400 mg/d therapy, respectively. In ENEST1st (NCT01061177), 38.6% of pts achieved MR4.5 after 24 mo of NIL 2*300 mg/d therapy. Methods: The ENESTfreedom (NCT01784068) study prospectively analyzed the stability of TFR in CP-CML pts after 1st line NIL therapy. Eligible pts had typical b2/3a2 BCR-ABL transcripts, ≥2 y frontline NIL and had achieved MR4.5. On study, pts continued NIL for 1 y; pts meeting predefined response criteria during the consolidation phase were eligible to stop NIL (TFR phase). NIL reinitiation was triggered by loss of major MR (MMR [BCR-ABLIS ≤0.1%]). Results: 215 pts (50 pts from 27 sites in Germany and Austria) were enrolled and entered the consolidation phase; 190 of these pts stopped NIL. Median age was 55 y, and median NIL duration prior to TFR was 43 mo (range, 33-89 mo). At wk 48 of the TFR phase, 51.6% (95% CI, 44.2-58.9%) of 190 pts remained in MMR, and 47.4% (95% CI, 40.154.7%) were in MR4.5 without reinitiation of treatment. A total of 86 pts lost MMR; 85 (98.8%) and 76 (88.4%) pts regained MMR and MR4.5 after NIL reinitiation, respectively. After NIL discontinuation, 47 pts (24.7%) reported musculoskeletal pain. Conclusions: Rate of DMR is significantly improved with 1st line NIL vs imatinib therapy. In ENESTfreedom, >50% of pts remained in TFR 48 wk after stopping NIL, a clinically meaningful rate for pts with a relatively short duration of NIL therapy (median 3.6 y). To test the impact of immune response for TFR, the ongoing TIGER trial (NCT01657604) investigates (i) the rate of molecular response after NIL vs NIL+PEG-Interferon (IFN) therapy and (ii) the stability of DMR after NIL vs NIL+PEG-IFN

Abstracts

CONTENTS AUTHOR INDEX

and PEG-IFN maintenance therapy. So far, 530 pts have been recruited from 123 sites in Germany, Switzerland and the Czech Republic. Disclosure: Ekkehard Eigendorff: Financing of Scientific Research: Novartis, BMS, Pfizer, Ariad Andreas Hochhaus: Financing of Scientific Research: Novartis, BMS, Pfizer, Ariad; Expert Testimony: Novartis, BMS, Pfizer, Ariad V154

Treatment free remission (TFR) in CML patients after 2nd line nilotinib (NI) therapy Dengler J.1, Stegelmann F.2, Sauer A.3, Saußele S.4, le Coutre P.5 Onkologische Schwerpunktpraxis Heilbronn, Heilbronn, Germany, Department of Internal Medicine III, University Hospital Ulm, Ulm, Germany, 3 Medizinisches Versorgungszentrum für Blut- und Krebserkrankungen, Potsdam, Germany, 4II. Medizinische Klinik, Universitätsmedizin, Medizinische Fakultät Mannheim der Universität Heidelberg, Mannheim, Germany, 5 Medizinische Klinik mit Schwerpunkt Onkologie und Hämatologie, Campus Charité Mitte, Charité – Universitätsmedizin Berlin, Berlin, Germany 1 2

Introduction: Achievement of a sustained deep molecular response (DMR) is a key prerequisite for successful TFR following frontline tyrosine kinase inhibitor (TKI) therapy; however, limited data are available on TFR following 2nd-line TKI therapy. The single-arm phase 2 study ENESTop explores the discontinuation of NI therapy in pts who achieved sustained MR4.5 (BCR-ABL1 ≤ 0.0032% on international scale IS) after switching from imatinib (IM) to NI. To estimate the proportion of pts who may be eligible to attempt TFR following 2nd-line NI, recently shown 4-y data from ENESTcmr has been considered. Methods: In ENESTop CP-CML pts were enrolled with previous TKI therapy for ≥ 3y (including IM for > 4 wk followed by NIL for ≥ 2 y) and achieved MR4.5 on NI. On study, pts continued NI for 1y (consolidation phase CONS). After 1y, pts without confirmed loss of MR4.5 were eligible to stop NI. Primary endpoint was the proportion of pts with successful TFR (neither loss of MMR nor confirmed loss of MR4by 48 wk of TFR) after ≥ 48wk follow up. Results: In ENESTop, 126 of 163 pts (13 pts contributed from Germany) in CONS entered TFR (median duration of TKI prior to TFR, 87.7mo; median duration of NI, 53mo). At data cut-off 57.9% remained in TFR at 48 wk. During TFR, 18 pts had confirmed loss of MR4, 34 lost MMR. Of the 51 pts who re-initiated NI, 98% regained at least MMR by data cutoff, and 94.1% and 92.2% regained MR4 and MR4.5, respectively. Median time to regain MR4/MR4.5 was 12/13.1 wk. No new safety findings were observed on treatment. Conclusion: ENESTop provides to date the largest prospective TFR data set in pts who achieved a sustained DMR after switching from IM to NI. Transfer from ENESTcmr to ENESTop is limited due to limited follow-up in ENESTcmr study. Yet overall, these results suggest that for pts lacking DMR on frontline IM, switching to 2nd-line NI may provide a route to TFR eligibility after shorter treatment duration. Disclosure: Jolanta Dengler: Employment or Leadership Position: Selbstständige Ärztin; Financing of Scientific Research: Novartis Philipp le Coutre: Employment or Leadership Position: Angestellter Arzt; Financing of Scientific Research: Novartis, BMS, Ariad, Pfizer

V155

Treatment optimization of newly diagnosed BCR-ABL positive patients with chronic myeloid leukemia (CML) in chronic phase with nilotinib vs. nilotinib plus interferon α induction and nilotinib or interferon α maintenance therapy: Interim analysis of the TIGER (CML V) study Hochhaus A.1, Saussele S.2, Baerlocher G.M.3, Brümmendorf T.H.4, Burchert A.5, Eigendorff E.1, La Rosée P.6, Hasford J.7, Hehlmann R.2, Heim D.8, Krause S.W.9, Le Coutre P.10, Niederwieser D.11, Lange T.12, Schenk T.1, Fabisch C.1, Pfirrmann M.7, Ostdeutsche Studiengruppe Hämatologie und Onkologie (OSHO), Schweizerische Arbeitsgemeinschaft für Klinische Krebsforschung (SAKK), Deutsche CML-Studiengruppe Klinik für Innere Medizin II, Universitätsklinikum, Jena, Germany, III. Medizinische Klinik, Universitätsmedizin, Mannheim, Germany, 3 Universitätsklinik für Hämatologie und Hämatologisches Zentrallabor, Inselspital, Bern, Switzerland, 4Klinik für Onkologie, Hämatologie und Stammzelltransplantation, Universitätsklinikum, Aachen, Germany, 5Klinik für Hämatologie, Onkologie und Immunologie Universitätsklinikum Gießen und Marburg GmbH, Marburg, Germany, 6Klinik für Innere Medizin II, SchwarzwaldBaar Klinikum, Villingen-Schwenningen, Germany, 7Institut für Med. Informationsverarbeitung, Biometrie und Epidemiologie, Ludwig MaximiliansUniversität, München, Germany, 8Klinik für Hämatologie, Universitätsspital, Basel, Switzerland, 9Medizinische Klinik 5 - Hämatologie und Internist. Onkologie Universitätsklinikum, Erlangen, Germany, 10Medizinische Klinik m.S. Hämatologie und Onkologie, Charité, Berlin, Germany, 11Abt. Hämatologie/ Onkologie, Universitätsklinikum, Leipzig, Germany, 12Klinik für Hämatologie und internistische Onkologie, Asklepios-Klinikum, Weissenfels, Germany 1 2

Introduction: The TIGER study (NCT01657604) is a multicenter, randomized phase III trial to evaluate efficacy and tolerability of nilotinib (NIL) 2*300mg/d monotherapy vs NIL 2*300mg/d + pegylated interferon α2b (Peg-IFN) and the option to discontinue therapy after Peg-IFN maintenance. A pilot phase preceded the main phase to evaluate the tolerability of NIL 2*300mg/d combined with PEG-IFN (30-50µg/week) commenced after >6 weeks NIL monotherapy. Here, data from the 1st interim analysis according to the protocol are presented. Methods: Between 8/2012 and 1/2013, 25 patients (pts) were recruited for the pilot phase. During the main phase of the study, newly diagnosed pts are being randomized between NIL 2*300 mg/d and NIL/PEG-IFN combination as confirmed during the pilot phase. An interim analysis was performed for the initial 200 pts with a minimum of 6 mo therapy. Efficacy and safety data are presented without specification of the randomized treatment arm due to ongoing recruitment. Results: As of 4/2016, a total of 511 pts (300 male; median age 53, range 18-85 years; 13% EUTOS high risk) were recruited from 123 sites in Germany, Switzerland, and the Czech Republic. The pilot phase of 25 pts confirmed the feasibility of the combination of NIL 2*300mg/d and PEGIFN 30µg/week. With regard to efficacy, 143/183 pts (73%) reached MMR (BCR-ABL1 transcripts < 0.1% according to the international scale) at 12 mo, 141/158 pts (89%) at 18 mo, and 85/89 pts (96%) at 24 mo. 63 pts concluded the induction phase and reached the maintenance phase of the study. Probabilities of adverse events grade 3-5 after 6 mo. of therapy are 21 and 26% for the two treatment arms. Five pts progressed to accelerated phase or blast crisis; one of them died from blast crisis, 2 received an allogeneic stem cell transplantation. Of 4 death total, 2 were CML related, one resulted from vascular complications, and one from an accident. Conclusions: Our data demonstrate feasibility of 1st-line treatment with NIL 2*300 mg/d combined with PEG-IFN 30-50 µg/week commenced after >6 weeks of NIL therapy. Molecular response during the first 24 mo compares favorably with data from recent studies and allows access to the maintenance phase (NIL vs PEG-IFN monotherapy) for the majority of patients. Randomization will be continued until the protocol defined recruitment goal (n = 652) will have been reached. Disclosure: Andreas Hochhaus: Financing of Scientific Research: Novartis, BMS, Pfizer, Ariad; Expert Testimony: Novartis, BMS, Pfizer, Ariad Markus Pfirrmann: No conflict of interest disclosed.

Abstracts

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CONTENTS AUTHOR INDEX

V156

V157

Expression of the CTLA-4 ligand CD86 (B7.2) on plasmacytoid dendritic cells (pDC) predicts risk of disease recurrence after treatment discontinuation in chronic myeloid leukemia

Deregulated CD62L expression predicts molecular response to nilotinib therapy in early chronic phase Chronic Myelogenous Leukemia (CML-CP)

Burchert A.1, Inselmann S.1, Saußele S.2, Dietz C.T.2, Müller M.C.2, Eigendorff E.3, Brendel C.A.1, Metzelder S.K.1, Brümmendorf T.H.4, Waller C.5, Dengler J.6, Goebeler M.E.7, Herbst R.8, Freunek G.9, Stefan H.10, Illmer T.11, Wang Y.1, Lange T.12, Finkernagel F.13, Hehlmann R.2, Huber M.14, Neubauer A.1, Hochhaus A.3, Guilhot J.15, Mahon F.X.16, Pfirrmann M.17, Schütz C.1, CML-Studiengruppe

Wolf D.1, Mustjoki S.2, Gjertsen B.-T.3, Gastl G.4, Baldauf M.5, Trajanoski Z.5, Giles F.6, Hochhaus A.7, Ernst T.7, Schenk T.7, Janssen J.8, Porkka K.9, Sopper S.4

Universitätsklinikum Marburg, Marburg, Germany, 2Universität Heidelberg, Universitätsmedizin Mannheim, Mannheim, Germany, 3Universitätsklinikum Jena, Jena, Germany, 4Universitätsklinik der RWTH Aachen, Aachen, Germany, 5 Universitätsklinik Freiburg, Freiburg, Germany, 6Onkologische Praxis Heilbronn, Heilbronn, Germany, 7University Hospital Würzburg, Würzburg, Germany, 8 Klinikum Chemnitz, Chemnitz, Germany, 9MVZ Klinikum Straubing GmbH, Straubing, Germany, 10Klinikum Kempten-Oberallgäu GmbH, Kempten, Germany, 11Gemeinschaftspraxis für Hämatologie und Onkologie, Dresden, Germany, 12Asklepios Klinikum Weißenfels, Weißenfels, Germany, 13Zentrum für Tumor- und Immunbiologie, Marburg, Germany, 14Biomedical Research Center (BMFZ), Marburg, Germany, 15Clinical Investigation Center, Poitiers, France, 16 Hematology Laboratory, CHU Bordeaux, Bordeaux, France, 17IMBE, LMU München, München, Germany 1

Introduction: It is unclear, why only a minority of chronic myeloid leukaemia (CML) patients eventually sustains molecular remission after discontinuation of tyrosine kinase inhibitor (TKI) therapy in deep molecular remission. Within the EURO-SKI TKI discontinuation study (EUDRACT 2011-000440-22) we studied expression of the CTLA-4 immune checkpoint ligand and co-stimulatory molecule CD86 (B7.2) on BDCA2+/ CD123+ plasmacytoid dendritic cells (pDC) using flow cytometry. We asked, whether CD86 expression on these specialized, interferon producing dendritic cells governs relapse risk after TKI stop. Methods: Peripheral blood samples of 122 patients were analysed before TKI treatment discontinuation. The expression of CD86 on BDCA2+/ CD123+ pDC and PD-1 on CD8+ T-cells, and CD8+ proteinase-3 specific T-cells was studied. Molecular relapse in this study was defined as loss of major moleuclar remission (loss of MMR). Results: The 1-year molecular relapse-free survival (RFS) was 30.1% (95% confidence interval [CI] 15.6-47.9) for patients with > 95 CD86+pDC / 105 lymphocytes prior to TKI stop versus 70.0% (95% CI 59.3-78.3) for patients with < 95 CD86+pDC (hazard ratio [HR] 3.4, 95%-CI: 1.9-6.0; p < 0.0001). The RFS for patients with < 95 CD86+pDC was significantly better in case of > 8 years TKI exposure before discontinuation (HR 0.3, 95% CI 0.1-0.8; p = 0.0263), whereas there was no apparent benefit from longer TKI pre-treatment with > 95 CD86+pDC. High CD86+pDC counts significantly correlated with leukaemia specific T-cell exhaustion (Spearman correlation: 0.74, 95%-CI: 0.21-0.92; p = 0.0098). Conclusion: CD86+pDC counts predict relapse risk of CML patients discontinuing a TKI in deep molecular remission. Only patients with low CD86+pDC counts seem to benefit from long TKI treatment duration before TKI cessation. Blocking CTLA-4 and PD-1 using immune checkpoint inhibitors could be a new treatment strategy to improve RFS in CML. Disclosure: Andreas Burchert: Financing of Scientific Research: BMS, Pfizer Christin Schütz: No conflict of interest disclosed.

Universitätsklinikum Bonn, Bonn, Germany, 2Hematology Research Unit Helsinki, University of Helsinki and Department of Hematology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland, 3Institute of Medicine, Hematology Section, University of Bergen, Bergen, Norway, 4 Department of Hematology and Oncology, Innsbruck Medical University, Innsbruck, Austria, 5Divison for Bioinformatics, Biocenter, Innsbruck Medical University, Innsbruck, Austria, 6NMDTI, Northwestern University, Chicago, United States, 7Klinik für Innere Medizin II, Universitätsklinikum Jena, Jena, Germany, 8Department of Hematology, VU University Medical Center, Amsterdam, Netherlands, 9Hematology Research Unit Helsinki, University of Helsinki and Department of Hematology, Helsinki, Finland 1

Introduction: Immunological surveillance of minimal residual disease (MRD) in chronic myelogenous leukemia (CML) may relevant for longterm control or cure of the disease. Little is known about immunomodulatory effects of nilotinib in vivo potentially predicting response to therapy. Methods: A prospective and comprehensive flow cytometry-based immunomonitoring program paralleled the ENEST1st clinical study (NCT01061177), investigating 52 nilotinib-naïve CP-CML patients and after treatment with 300 mg BID nilotinib at months 6 and 12. Molecular data were analyzed in central EUTOS reference laboratories. The data were verified in an independent validation cohort (n = 21) from the CML IV TIGER trial. Results: T cells of CML patients at diagnosis displayed an aberrant phenotype characterized by very low L-selectin (CD62L) expression levels, which was not due to proportional shifts from CD62Lhigh to CD62Llow or negative T cell subsets. At diagnosis, low numbers of CD62L-expressing CD4+ and CD8+ T cells highly correlated with important baseline disease characteristics, such as higher SOKAL score, increased spleen size and high leukocyte as well as peripheral blood blast counts. Six months after nilotinib-therapy initiation, expression of CD62L returned back to levels seen in healthy individuals. Of note, the amount of reduced CD62L expression on T cells directly correlated with the extent of soluble CD62L (sCD62L) elevation in the plasma of diagnostic CML patients, which returned back to normal levels at months 6 and 12 during CML therapy. In parallel to the increase of sCD62L at diagnosis, the proteolytic activity but not the total amount of TACE (ADAM17, CD156b), the metalloproteinase known to shed CD62L is clearly increased at diagnosis and significantly decreased during nilotinib therapy. High CD62L expression on both CD4+ and CD8+ T cells and vice versa low sCD62L levels at CML diagnosis were linked to superior molecular responses (MMR, MR4 and MR4.5). These findings were corroborated in the independent validation cohort from the german CML IV TIGER trial. Conclusions: We demonstrate for the first time the prognostic impact of increased CD62L shedding from T cells at CML diagnosis for later molecular response to nilotinib in early CML-CP. Decreased CD62L is most likely mediated by increased TACE-activity. This may impair T cell-mediated leukemia control, as CD62L is important for T cell entry to secondary lymphoid organs. Disclosure: Dominik Wolf: Advisory Role: Novartis; Financing of Scientific Research: Novartis; Expert Testimony: Novartis Sieghart Sopper: Expert Testimony: Novartis

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V158

The predictive value of early molecular response by relative risk in patients (pts) with chronic myeloid leukemia in chronic phase (CML-CP) treated with dasatinib (DAS) or imatinib (IM) from the phase 3 DASISION trial Stegelmann F.1, Shah N.P.2, Saglio G.3, Hochhaus A.4, Bilmes R.5, Li L.5, Cortes J.E.6 Universitätsklinikum Ulm, Ulm, Germany, 2UCSF School of Medicine, San Francisco, United States, 3University of Turin, Turin, Italy, 4Universitätsklinikum Jena, Jena, Germany, 5Bristol-Myers Squibb, Princeton, United States, 6University of Texas M.D. Anderson Cancer Center, Houston, United States 1

Introduction: Improved outcomes were observed in treatment-naive CMLCP pts treated with DAS or IM who achieved the early molecular milestone of BCR-ABL1 ≤10% at 3 mo in an exploratory landmark analysis of the phase 3 DASISION trial, and BCR-ABL1 ≤10% at 3 mo was achieved by more pts on DAS (DAS: 84%, IM: 64%). We therefore evaluated the impact of EURO (Hasford) risk score on the achievement of this predictive molecular milestone and investigated other predictive factors of survival. Methods: In DASISION, newly diagnosed CML-CP pts were randomized to receive DAS 100 mg once daily (n = 259) or IM 400 mg once daily (n = 260). BCR-ABL1 transcript levels were measured on the International Scale. EURO score ≤780 is low, >780 to ≤1480 is intermediate, and >1480 is high risk. Results: Based on EURO score, 33% of pts in both treatment arms were at low risk, 48% and 47% of pts receiving DAS and IM, respectively, were at intermediate risk, and 19% in both arms were at high risk. Across all risk groups, more pts on DAS had BCR-ABL1 ≤10% at 3 and 6 mo vs IM (3 mo: low 91% vs 73%, intermediate 80% vs 66%, high 83% vs 44%; 6 mo: low 99% vs 89%, intermediate 84% vs 82%, high 84% vs 71%). More pts on DAS vs IM also had BCR-ABL1 ≤1% at 3 mo (low 56% vs 20%, intermediate 48% vs 12%, high 33% vs 5%) and at 6 mo (low 79% vs 58%, intermediate 66% vs 50%, high 60% vs 29%). Five-y molecular responses and outcomes were higher for pts with BCR-ABL1 ≤10% vs >10% at 3 mo in all risk groups (Table), as were 5-y response and outcomes for pts with BCR-ABL1 ≤10% vs >10% at 6 mo in all risk groups. Multivariate analysis of survival at 5 y showed women on DAS were at a lower risk of death than men; there was no association between EURO score or age and survival. Conclusions: More pts achieved BCR-ABL1 ≤10% at 3 and 6 mo on DAS vs IM regardless of EURO risk score, with a significantly higher proportion of pts on DAS with BCR-ABL1 ≤1%. The largest difference in BCRABL1 levels between the treatment arms was observed in those at high risk. BCR-ABL1 ≤10% vs >10% at 3 and 6 mo was predictive of improved long-term outcomes in all risk groups. Tab. 1. DGHO Table

Wissenschaftliches Symposium Neue Optionen beim Hodgkin-Lymphom V160

PET guided therapy of Hodgkin Lymphoma Borchmann P.1 Uniklinik Köln, Klinik I für Innere Medizin, Köln, Germany

Treatment with current radio- and chemotherapy regimens cures the majority of patients suffering from Hodgkin lymphoma (HL). Treatment allocation is based on the Ann-Arbor system, which mainly reflects the burden of disease. Within the three risk groups (early, intermediate, and advanced stage HL), the individual HL patients still may have very different risk profiles. Reliable predictive markers at baseline are not available. Therefore, current standard therapies such as ABVD (doxorubicin, bleomycin, vinblastine, and dacarbazine), or BEACOPP (bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, prednisone) are administered to all patients with newly diagnosed HL without knowing the individual risk for treatment failure. Consequently, over- and undertreatment may occur and may result in either excessive toxicity or preventable relapse. To overcome these limitations regarding the individual upfront risk assessment, the concept of response-adapted therapy has been developed. According to this concept, treatment intensity is either de-escalated or intensified depending on the observed early response to therapy. This more individualized treatment approach has become possible by the introduction of functional imaging using 18F-fluoro-2-deoxy-D-glucose positron emission tomography (18FDG PET) for response assessment. Determining the metabolic response to therapy seems to generate more prognostic information than the radiological assessment alone. This concept has been tested in all different stages of HL in large phase III studies. Results have become available in the last few years and will be summarized. Disclosure: No conflict of interest disclosed.

Fortbildung

Standardisierte Leukämie-Diagnostik V168

Next-generation sequencing Haferlach T.1 MLL Munich Leukemia Laboratory, München, Germany

Disclosure: Frank Stegelmann: Advisory Role: ARIAD, PFIZER; Honoraria: NON; Financing of Scientific Research: ARIAD, BMS, NOVARTIS, PFIZER; Other Financial Relationships: BMS, NOVARTIS (travel expenses for scientific meetings) Jorge E. Cortes: Advisory Role: Ariad, BMS, Novartis, Pfizer; Expert Testimony: Ariad, BMS, Novartis, Pfizer, Teva

The recent ten years demonstrated an outstanding success for molecular approaches in hematology: This is true for research discoveries, to define markers at diagnosis, for prognostication, for follow-up studies including minimal residual disease and also for target detection. This has been possible due to ground breaking developments in techniques such as NGS, instruments, automation and also in software tools, to make data appropriate for science as well as for routine diagnostic purposes. Several techniques have been introduced to the market, some only for research, some also with FDA or CE market labels, accredited for routine diagnostic purposes. Allover, the turnaround time decreased, the specificity and the sensitivity increased, assays included the investigation of single genes, gene panels, but also whole exome sequencing and whole genome sequencing is now possible. In parallel, costs are going down, making it even possible to implement next generation sequencing not only for highly skilled research institutes but also for routine laboratory investigations. Although the recent technological opportunities with respect to assay development, data read-out and turnaround time is impressive, the most challenging field so far is to interpret the data correctly for any research project but especially for report generation in patients care. Several international available databases in the www have to be addressed in parallel

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

CONTENTS AUTHOR INDEX

to interpret the respective findings on patients DNA or RNA. In parallel, an in-house database should be developed and curated. As the recent opportunities also lead to findings that could not be clearly interpreted as being damaging (malignant) or benign but maybe so-called a “variant of unknown significance” (VUS), the management of data today is more complex than the technique NGS itself. Several important approaches, also guided by the FDA and the ASH including the task force for precision medicine are working in the field of standardization, accreditation and data management in the upcoming era of NGS driven molecular diagnostics. However, the options are opening a new field not only to understand the pathobiology of hematology but also to foster diagnostic approaches and lead more and more directly to targeted treatment. Disclosure: Torsten Haferlach: Employment or Leadership Position: MLL Muich Leukemia Laboratory V169

Development and application of WHO standards for standardization of molecular monitoring in CML Cross N.C.P.

University of Southampton, Salisbury, United Kingdom

Serial quantification of BCR-ABL1 mRNA levels is an important therapeutic indicator for patients with chronic myeloid leukaemia, but historically there has been substantial variation in results reported by different laboratories. To help improve the comparability of results, an International Scale (IS) for BCR-ABL1 has been developed and implemented by many testing laboratories worldwide, initially by the derivation of laboratory-specific conversion factors (CFs) following sample exchange with a reference laboratory. Whilst the development of CFs was a major step forward and provided an important proof of principle, it is obvious that this approach is not sustainable in the long term. Ideally, any testing laboratory should be able to access reference standards or use a kit that enables them to convert patient results directly to the IS. The development of standards and kits initially required the development of a process by which these tools could be calibrated to the IS. An important milestone in this process was the establishment in 2010 of the 1st World Health Organization International Genetic Reference Panel for quantitation of BCR-ABL1 mRNA. The reference panel comprises four different dilution levels of freeze dried preparations of K562 cells diluted in HL60 cells that were assigned fixed % BCR-ABL1/reference gene values on the IS following an international calibration process. Due to the scale of molecular monitoring, it was not physically possible to manufacture and validate a sufficiently large quantity of reference material to satisfy worldwide demand and thus the principal function of these primary reagents was limited to the calibration of secondary reference reagents. These secondary reference reagents may be manufactured and calibrated by companies, reference laboratories or other agencies and made available to testing laboratories either on a commercial basis or as part of specific national or regional standardization initiatives. Several different kits, systems and secondary reagents are available that enable testing laboratories to derive patient results on the IS. Comparative data is, however, very limited at present and it is not possible to say which of these approaches is best. It is clear, however, that use of an IS kit does not guarantee accurate results and additional quality control measures are required. Disclosure: Nicholas Cross: Financing of Scientific Research: Novartis; Expert Testimony: Novartis

Oncol Res Treat 2016;39(suppl 3):1–104

Freier Vortrag

Versorgungsforschung V170

Use of local oncological Standard Operating Procedures (SOPs) and oncological guidelines: A survey of 1600 physicians at 4 oncology sites/centers of excellence Krebs S.1, Starbatty B.1, Skoetz N.1, Schmidt-Wolf I.2, Brandts C.3, Bischoff M.4, Wolf J.1, Glossmann J.P.1, Centers of excellence working group SOP Universitätsklinikum Köln, Centrum für Integrierte Onkologie (CIO) Köln Bonn, Köln, Germany, 2Universitätsklinikum Bonn, Centrum für Integrierte Onkologie (CIO) Köln Bonn, Bonn, Germany, 3Universitätsklinikum Frankfurt, Universitäres Centrum für Tumorerkrankungen (UCT), Frankfurt am Main, Germany, 4 Universitätsklinikum Freiburg, Department für Medizinische Biometrie und Medizinische Informatik, Freiburg i. Br., Germany 1

Introduction: All centers of excellence in Germany are required to develop and provide standards for cancer care and treatment. The center of excellence network, funded by the German Cancer Aid, established a working group for SOPs. Objectives of this group are to analyze SOPs in the centers to standardize content and structure of SOPs, to develop new SOPs for rare cancers and to give impetus to the evidence based national S3-Guidelines. While guidelines of the medical societies are nationally important and build the base for SOPs, the SOPs also implement local specific conditions and circumstances of the center. Methods: We developed a survey for physicians practicing in the centers of excellence to assess knowledge and usage of SOPs. A pretest of the survey was conducted, involving ten physicians. The final version was approved by the Employee Committees, Data Protection Supervisors and ethics committees. The survey was web-based and anonymous and conducted in the centers Cologne, Bonn, Frankfurt and Freiburg. We assessed baseline characteristics as well as questions related to local web-based SOPs and national S3-guidelines. Only datasets with at least 30% of the survey completed were evaluated. Results: 1669 doctors were invited. 1175 responsed and 424 responses reached the ≥30% completion limit. 60% were male, 40% female. 6% are chief physicians, 33% consultant physicians, 18% board certification and 43% assistant physicians. Tab. 1. Knowledge SOP-Portal

Do you know that SOPs for oncological diseases are available in the local web-based SOP portal? Yes No

(%) 64 36

Tab. 2. Topics

Which topics are of most interest for you? Diagnostics Stage/Classification Therapy Follow up

… in SOPs (%) … in guidelines (%) 52 51 80 42

57 56 80 39

… SOPs (%)? 20 34 21 11 5 9

… guidelines (%)? 43 22 17 3 6 9

Tab. 3. Frequency

How often do you use Weekly Once a month Every three months Once a year Less than once a year Never

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CONTENTS AUTHOR INDEX

Conclusion: 64% of physicians are familiar with the SOPs. Rating of the topics importance is similar for SOPs and guidelines. Therapeutic aspects are the most important information. According to frequency of usage, oncological guidelines seem to be used more regulary compared to SOPs. Disclosure: No conflict of interest disclosed. V171

Systemic palliative treatment in patients with metastatic colorectal cancer tested for their (K)RAS mutation status – Real-life data from the clinical Tumour Registry Colorectal Cancer (TKK) Zahn M.O.1, Schnell R.2, Karcher A.3, Schnitzler M.4, Wetzel N.4, Fleitz A.4, Marschner N.5, for the TKK registry group Studienzentrum MVZ Onkologische Kooperation Harz, Goslar, Germany, Praxis Internistischer Onkologie und Hämatologie (PIOH), Frechen, Germany, 3 Onkologische Schwerpunktpraxis, Heidelberg, Germany, 4iOMEDICO, Freiburg, Germany, 5Praxis für Interdisziplinäre Onkologie & Hämatologie, Freiburg, Germany 1 2

Background: RAS has been identified as a predictive biomarker for treatment response and outcome in patients (pts) with metastatic colorectal cancer (mCRC). Pts with RAS mutation (mt) do not benefit from treatment with EGFR-inhibitors. Since 2008 KRAS mutation testing was mandatory before applying the EGFR-inhibitors cetuximab (CET) and panitumumab (PAN). Since 2014 expanded RAS mutation testing is required. Pts with RAS mt can be treated with chemotherapy (CT), with or without the VEGF-inhibitor bevacizumab (BEV). A VEGF- or EGFR-inhibitor in combination with CT or as single agent is recommended for RAS wildtype (wt) pts. Data from recent studies indicate a better survival of pts with RAS wt treated with a regimen with an EGFR-inhibitor in 1st-line and a regimen with a VEGF-inhibitor in 2nd-line. Methods: Since 2006, the Tumour Registry Colorectal Cancer (TKK) prospectively documents treatment of pts with mCRC by medical oncologists in Germany. Pts receive treatments according to physician’s choice. The TKK collects data on all systemic treatments, biomarker testing, pts and tumour characteristics and outcome. By March 2015, 166 sites recruited 3500 mCRC pts. Data on mutation status, referred to as (K)RAS mutation status, has been collected for KRAS since 2008 (n = 2264) and for RAS since 2014 (n = 226). Results: The number of pts tested for KRAS increased from 52% in 2008 to 72% in 2013. In 2015 79% of pts have been tested for extended RAS mt status. 66% of pts were tested with KRAS wt until 2013. With expanded RAS mutation testing, this number decreased to 48% of pts tested with RAS wt in 2014–15. In pts with (K)RAS wt the most frequently applied 1st-line regimens from 2010–2013 were FOLFIRI or FOLFOX, with or without BEV or CET. The proportion of pts treated with CET was 33%. In 2014–15 the use of a regimen with CET or PAN increased to 48% with FOLFIRI+CET and FOLFOX+PAN as the two most frequently applied regimens. In 2nd-line treatment of (K)RAS wt pts the use of FOLFIRI+CET decreased from 2010–2015, while treatment with FOLFIRI+BEV increased and is the most frequently applied regimen in 2014/15. In pts with (K)RAS mt the most frequently used regimens were FOLFIRI+BEV, FOLFOX+BEV, FOLFOX and FOLFIRI in 1st- and 2nd-line. Conclusions: In 2014/15 the majority of pts were tested for RAS mutation status before the start of 1st-line therapy. Upcoming analyses will show how recent study results on treatment with EGFR-VEGF sequence will influence routine treatment. Disclosure: No conflict of interest disclosed.

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V172

Oncologists´ consultation: Attitudes of migrant patients Riese C.1, Borges jr. U.1, Mödder M.1, Baumann W.1 Wissenschaftliches Institut der Niedergelassenen Hämatologen und Onkologen – WINHO, Köln, Germany 1

Background: Since one in five residents in Germany have a migration background, we focused on the implications of language barriers and cultural differences for the patients’ comprehension of treatment and on their attitudes concerning cancer care. Differences between patients with and without migration experience should be considered. Methods: The WINHO patient survey is held annually and serves as benchmarking for the participating office-based oncology practices as well as baseline study for specific topics. In 2015, the participants were surveyed in: a) perception of the practice structure and conversation with the doctor, b) communication and attitudes, c) sociodemographic questions. The answers of patients with migration experience were compared to those born in Germany. Group differences were tested for significance via χ²-test. Results: 9540 patients participated (return rate: 84%; 53.3% female, M = 64.93 years, SD = 13.2). 858 (9%) were not born in Germany. Migrants with a lower command of the German language more often reported the impression that crucial parts of the conversation with the oncologist remained incomprehensible (27.7%) than migrants with good knowledge of German (9.2%) and native German speakers (5.8%), p < .001. The understanding of written information relevant for treatment provided by practices was more often difficult for migrants with less knowledge of German than for those with good knowledge and for native speakers (respectively 10.4%, 4.1% and 2%), p < .001. Regarding expectations of treatment, foreign patients in general have a lower tendency to a) tell doctors about their needs and concerns regarding their treatment (89.3% compared to 94.2% of patients born in Germany), p < .01, b) tell their opinion on the intentions of the doctor (81.4% to 87.3%), p < .01, and c) desire to participate in crucial decisions regarding diagnosis and treatment (85.8% to 91.6%), p < .01. Conclusions: Survey participants with migration experience, especially those with less language proficiency, stated more difficulties with understanding of both written information and the verbal explanation by the oncologist during a consultation. As foreign patients with different cultural backgrounds are a very heterogeneous group, such differences need to be taken into account in further analyses. Disclosure: No conflict of interest disclosed. V173

Sensorimotor vs. endurance exercise in cancer survivors with chemotherapy-induced peripheral neuropathy Müller J.1, Wehrle A.2, Kneis S.1, Gollhofer A.3, Bertz H.1 Medical Center – University of Freiburg, Department Medicine I, Haematology, Oncology and Stem Cell Transplantation, Freiburg, Germany, 2Medical Center – University of Freiburg, Institute for Exercise- und Occupational Medicine, Center for Medicine, Freiburg, Germany, 3Department of Sport and Sport Science, Albert-Ludwigs-University Freiburg, Freiburg, Germany 1

Introduction: Symptoms of peripheral neuropathy often occur after treatment with neurotoxic chemotherapy, depending on agent and dose. Affected patients suffer from sensory and motor dysfunction resulting in limitations of their daily activities culminating in a necessary change of therapy regime. Up to now, there is no evident therapy to effectively reduce or prevent symptoms of chemotherapy-induced peripheral neuropathy (CIPN). Due to motor dysfunction in terms of balance problems, gait instability and risk of falling exercise seems to be a promising instrument to improve postural control in CIPN patients. Therefore, the aim of this study was to evaluate the relative benefit of sensorimotor exercise, which is known to improve neuromuscular function, versus endurance exercise, which is proven to reduce diabetes-induced neuropathic symptoms.

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Methods: 50 cancer patients after active treatment with persisting symptoms of CIPN were randomly allocated either to a sensorimotor exercise group (SG) or an endurance exercise group (EG). Both groups exercised three times per weeks over 12 weeks and underwent pre- and post-assessments: objective clinical tests and subjective symptom questionnaires to determine the level of CIPN, balance tasks to evaluate motor dysfunction and a maximum cardiopulmonary exercise test. Results: After intervention both groups reduced their subjective sensory symptoms significantly (SG: P = 0.028; EG: P = 0.037), while only SG reported also improved motor function (P = 0.006) measured by EORTC QLQ-CIPN20. Clinical tests revealed no significant changes. With increased difficulty of balance tasks, difference between groups appeared: SG reduced their sway path in semitandem stance significantly (P = 0.031) and improved their one leg stance time on instable surface (P = 001). Furthermore, cardiorespiratory fitness (VO2peak) descriptively increased in both groups, this change, however, did not reach statistical significance Conclusion: All patients benefited from exercising. However, the study revealed no clear superiority of sensorimotor training in terms of influencing motor dysfunction as hypothesized. Nonetheless patients of SG showed more positive changes than patients of EG. We speculated that the averaged expression of CIPN in our cohort was too weak and could therefore not be influenced by our interventions due to a ceiling effect. Further studies need to differentiate between moderate and severe CIPN to ensure a higher reliability. Grant: Janssen Cilag Disclosure: No conflict of interest disclosed. V174

Overall survival of 4865 patients with metastatic solid tumours treated in German routine practice – combined results from four prospective, multicentre cohort studies Marschner N.1, Potthoff K.2, Schnell R.3, Tesch H.4, Zahn M.-O.5, HegewischBecker S.6, Jänicke M.2, iOM-Registry group Praxis für interdisziplinäre Onkologie & Hämatologie, Freiburg i.Br., Germany, iOMEDICO, Freiburg i.Br., Germany, 3Praxis Internistischer Onkologie und Hämatologie (PIOH), Frechen, Germany, 4Onkologische Gemeinschaftspraxis, Frankfurt a.M., Germany, 5Studienzentrum MVZ Onkologische Kooperation Harz, Goslar, Germany, 6Onkol. Schwerpunktpraxis HH Eppendorf, Hamburg, Germany

Tab. 1. Patient characteristics and overall survival

Trial OS Age Comorbidity ineligible* (median (median) [%] [%] (95%-CI)) (K)RAS** 26.4 mCRC 1033 67 69% 19% wild-type (24.0–28.1) (K)RAS** 22.3 580 69 68% 26% mutation (20.5–23.9) HR positive 32.8 mBC 752 63 65% 14% Her2 negative (29.7–38.2) HR positive 43.0 301 62 61% 12% Her2 positive (33.8–53.7) HR negative 33.1 145 61 53% 22% Her2 positive (24.6–39.3) Triple 16.6 200 60 59% 14% negative (12.3–19.1) 10.8 mNSCLC 1403 66 79% 41% (9.8–11.6) 7.9 mPC 451 70 81% 21% (6.7–10.2) Tumour entity

* Pts were defined as “trial ineligible” when at least one of the exclusion criteria was met (compare text). Pts with missing values were excluded from this analysis. ** Data on mutation status has been collected for KRAS since 2008 and for RAS since 2014. KRAS/RAS status is summarised under the term (K)RAS. CI: Confidence Interval, HR: hormone receptor, mBC: metastatic breast cancer, mCRC: metastatic colorectal cancer, mNSCLC: metastatic non-small cell lung cancer, mPC: metastatic pancreatic cancer, OS: overall survival [months]

Conclusions: OS of pts with metastatic solid tumours in German routine clinical practice is similar to published data from randomized clinical trials although pts are older and most often affected with comorbidities. Disclosure: No conflict of interest disclosed.

1 2

Introduction: Published data on overall survival (OS) of patients (pts) with metastatic solid tumours are mostly based on randomized clinical trials conducted at university hospitals. In Germany, cancer care is predominantly delivered by office-based medical oncologists and oncologists based in community hospitals. They mainly treat cancer pts with various comorbidities generally excluded from clinical trials. Prospectively collected data on OS in the routine clinical setting are rare. Methods: Data source are prospective, multicentre, longitudinal cohort studies recruiting pts with metastatic colorectal (mCRC), breast (mBC), non-small cell lung (mNSCLC) or pancreatic (mPC) cancer. Pts were recruited at the start of first-line treatment by more than 250 study sites in Germany, mainly office-based medical oncologist practices and few community hospitals. Pts receive standard treatments according to physician’s choice. A broad set of data including pts and tumour characteristics, systemic treatments and outcome are collected. OS was estimated with the Kaplan-Meier method. Common clinical trial exclusion criteria were defined as ECOG≥2 or presence of renal disease, moderate or severe liver disease, heart failure or brain metastases at the start of first-line treatment (“trial ineligible”). Results: All pts started their first-line treatment between 2006 and 2015. Table 1 presents pts characteristics and OS. About 70–80% of pts had one or more relevant comorbidities, mainly hypertension. In mBC only 53–65% of pts had comorbidities. The percentage of pts who were trial ineligible ranges from 12% (HR/Her2 positive mBC) to 41% (mNSCLC). About 20% of pts with mCRC and mPC met at least one exclusion criterion of a clinical trial.

Oncol Res Treat 2016;39(suppl 3):1–104

V175

Recommendations for diagnosis, treatment and follow-up in oncology – an inventory of Standard Operating Procedures (SOPs) of centers of excellence Glossmann J.-P.1, Starbatty B.2, Bischoff M.3, Follmann M.4, Skoetz N.2, Centers of excellence working group SOP Centrum für Integrierte Onkologie (CIO) KölnBonn, Köln, Germany, 2Centrum für Integrierte Onkologie (CIO) KölnBonn, Koordinationsstelle der AG SOP, Köln, Germany, 3Uniklinik Freiburg, Institut für Medizinische Biometrie und Informatik, Freiburg, Germany, 4Leitlinienprogramm Onkologie der AWMF, DKG und DKH, Berlin, Germany

Abstracts

CONTENTS AUTHOR INDEX

As the first survey in 2012 identified differences in the actuality, methods used to develop SOPs, members of the working group developed recommendations how to prepare, update and implement a SOP. Results: In 2016, at least one SOP in the network exists for almost any kind of solid cancer (89 out of 90 ICD- codes), haematological malignancies and aspects of supportive care. This list of SOPs is available at www.ccc-netzwerk.de and interested centers or physicians may ask via der co-ordinating office (funded by the DKG, No. 111493) for access to these SOPs. The first network SOP thyroid cancer is currently in the final review process and will soon be freely available on this website. Moreover, recommendations how to prepare and update SOPs can be downloaded as a handbook, giving suggestions for harmonized methodology and evidence processing. One topic identified so far, without any published guidelines and SOPs, is familial cancers. The working group SOP has already started to co-ordinate these SOPs. Conclusions: SOPs are important for standardized diagnostic and treatment of patients and exist for almost all types of cancer in the network. Harmonized processes might increase the quality of cancer care throughout Germany. Disclosure: Jan-Peter Glossmann: Employment or Leadership Position: Sprecher AG SOP des Netzwerks onkologischer Spitzenzentren Nicole Skoetz: Employment or Leadership Position: Wissenschaftliche Koordinatorin AG SOP des Netzwerks onkologischer Spitzenzentren

Freier Vortrag

Akute myeloische Leukämie – Therapie 1, Mutationsprofil V178

Response-adapted sequential Azacitidine and induction chemotherapy in patients > 60 years with newly diagnosed AML eligible for chemotherapy: Interim analysis of the DRKS00004519 study (RAS-AZIC) Jäkel N.1, Hubert K.2, Krahl R.1, Hänel M.3, Maschmeyer G.4, Herbst R.3, Jacob C.4, Schulze S.1, Wang S.-Y.1, Cross M.1, Brosteanu O.5, Niederwieser D.1, Al-Ali H.K.1 Universitätsklinik Leipzig AöR, Hämatologie/ internistische Onkologie, Leipzig, Germany, 2Medizinische Fakultät/ Universität Leipzig, Hämatologie/ internistische Onkologie, Leipzig, Germany, 3Klinik für Innere Medizin III/ Klinikum Chemnitz gGmbH, Hämatologie, Onkologie, Stammzelltransplantation, Chemnitz, Germany, 4Klinikum Ernst von Bergmann gGmbH, Klinik für Hämatologie, Onkologie und Palliativmedizin, Potsdam, Germany, 5Universität Leipzig AöR, Koordinierungszentrum für Klinische Studien Leipzig, Leipzig, Germany 1

Treatment for AML in patients (pts) >60 years (y) includes intensive chemotherapy (IC) and/or azacitidine (AZA). Yet, AZA and IC need not be mutually exclusive. In this multicenter study of the East German Study Group, first-line treatment with AZA followed by AZA or IC in pts >60 y was evaluated. Data of a planned interim analysis are presented. Patients and methods: Patients >60y with newly diagnosed AML were included. Upfront AZA (75 mg/m2/day s.c.) for 7 days was followed by AZA or IC (mitoxantrone 10 mg/ m²/day on day (d)1–3 and cytarabine 1g/ m²/BID on d1, 3, 5, 7) based on d15 bone marrow (BM) blasts (< 45 versus ≥45%) and response on d56 previously identified as early predictors for response to AZA in AML. The primary endpoint was response (OR) [CR, CRi, PR] at d90 according to IWG criteria. Safety, OS, and TRM were secondary endpoints. On the basis of an optimal two-stage design, an expected OR of 61% at the end of the trial was estimated. Thus, if ≤ 19 of the first 40 pts did not achieve the OR, protocol treatment would be considered inferior to standard IC. All pts gave written informed consent. Results: The median age of the 40 pts was 70y (55% males). de novo AML was present in 63% of pts. Median BM blasts was 39%. Cytogenetics was high- in 29% and intermediate-risk in 71% of 38 pts. FLT3 and NPM1 were mutated in 11% and 17% of 36 pts respectively. All pts received up-

Abstracts

front AZA. 37 (92.5%) pts received protocol assigned treatment based on BM blasts on d15 (54% continued with AZA; 46% received IC). Overall, in the 33 (82.5%) pts who continued therapy per protocol until d90, an OR was achieved in 27 (82%) pts [CR/CRi (n = 17/7); PR (n = 3)]. In the intention-to-treat cohort (n = 40), this represents an OR of 67.5% [CR/ CRi (60%); PR (7.5%)] with AZA + one IC cycle (n = 28) or two IC cycles (n = 3). TRM on d30 and d90 was 0% and 5% respectively. After a median follow-up of 202 days, OS was 84.5%. OS for responding pts was 95.5%. Upfront AZA was well tolerated. Constipation grade 1+2 was the most frequently reported AE under AZA (45%). Overall, the most frequent grade 3+4 non-hematologic AE was infection [11 times in 31 IC-cycles (35%); 7 times in 60 AZA-cycles (12%)]. Conclusions: Response-based sequential epigenetic therapy and intensive chemotherapy is feasible in elderly pts with AML with a very low treatment-related mortality. The final results of the trial will scrutinize the impact of this approach on survival. Disclosure: Nadja Jäkel: No conflict of interest disclosed. Haifa Al-Ali: Financing of Scientific Research: yes; Expert Testimony: yes V179

Only SETBP1 hotspot mutations in myeloid neoplasm are associated with refractory disease and reduced overall survival- a single center analysis of 442 patients Winkelmann N.1, Schäfer V.1, Reichert J.1, Waldau A.1, Rinke J.1, Hochhaus A.1, Ernst T.1 Klinik für Innere Medizin II, Universitätsklinikum Jena, Hämatologie und internistische Onkologie, Jena, Germany 1

Introduction: SKI-homology domain aberrations of SETBP1 have been characterized as a diagnostic biomarker in myeloid neoplasm and are associated with inferior survival. Since there is limited data on the clinical impact of these aberrations and their stability during disease progression, we aimed to follow up and study affected patients from our institution. Methods: Bidirectional Sanger sequencing of gDNA from 442 unselected patients with WHO-defined myeloid disease was performed at diagnosis or first encounter (AML; n = 61, sAML; n = 117, MDS; n = 98, MPN; n = 111, MDS/MPN; n = 43, aplastic anemia; n = 12). Secondly, we conducted a follow up analysis on 190 samples from 123 patients at a median of 8 months (range: 1–62 months) from first screening. Results: Ten patients (2.3%) had aberrations in the highly conserved 11-nucleotide region of codons 868 to 871 (MDS/MPN, n = 6, sAML, n = 3, AML, n = 1). Germline material was available in 5/10 patients and confirmed the somatic origin of mutations. Five patients (1.1%) showed SETBP1 aberrations that were spread within the SETBP1 SKI homology domain (MPN, n = 1, MDS/MPN, n = 1, MDS, n = 2, sAML, n = 1). In three patients, germline material was available and showed the same aberration, indicating a polymorphism. These patients (median age: 64 ys) received demethylating therapy (n = 1) or supportive care (n = 4) and had a median overall survival of 34 months. The 10 patients with hotspot mutations (median age: 65 ys) underwent leukemia induction therapy (n = 1), allogeneic transplantation (n = 4) or palliative care (n = 6) and had a median overall survival of 4.5 months. All 3 transplanted patients had an early relapse after 2, 6 and 14 months, respectively. In 2/3 patients, the clone carrying the SETBP1 mutation was detectable by sanger sequencing at relapse. In univariate analysis of the entire cohort patients harboring SETBP1 hotspot mutations were significantly associated with unfavorable overall survival (4.5 vs. 14 months, p = 0.009). Targeted sequencing on cooperating mutations of patients with and without SETBP1 hotspot mutations is currently ongoing. Conclusions: Patients with SETBP1 hotspot mutations suffer from aggressive disease with rapid evolution and relapse after allo SCT. Diagnostic screens for SETBP1 hotspot mutations may help identifying this dismal patient group and treat them in multicenter clinical studies. With emerging data on the biologic function of these mutations, a targeted therapy may be available in the future. Disclosure: No conflict of interest disclosed.

Oncol Res Treat 2016;39(suppl 3):1–104

CONTENTS AUTHOR INDEX

V180

V181

Individual drug exposure to clofarabine measured by AUC predicts liver toxicity in patients with relapsed or refractory acute myeloid leukemia

Mutational profiling of AML patients with CEBPA-mutations reveals differences between biallelic and monoallelic TAD and bZIP mutant subgroups

Büttner B.1, Middeke J.M.2, Knoth H.1, Oertel R.3, Seeling A.4, Kramer M.2, Sockel K.2, von Bonin M.2,5,6, Stölzel F.2, Alakel N.2, Platzbecker U.2, Röllig C.2, Ehninger G.2, Bornhäuser M.2, Schetelig J.2,7

Georgi J.-A.1, Taube F.1, Kramer M.1, Herold S.1, Röllig C.1, Eberlein C.1, Stasik S.1, Krämer A.2, Berdel W.E.3, Serve H.4, Platzbecker U.1, Bornhäuser M.1, Ehninger G.1, Schetelig J.1, Thiede C.1, Studienallianz Leukämie (SAL)

Universitätsklinikum Carl Gustav Carus der TU Dresden, Klinikapotheke, Dresden, Germany, 2Universitätsklinikum Carl Gustav Carus der TU Dresden, Medizinische Klinik und Poliklinik I, Dresden, Germany, 3Medizinische Fakultät Carl Gustav Carus der TU Dresden, Institut für Klinische Pharmakologie, Dresden, Germany, 4Friedrich-Schiller-Universität Jena, Institut für Pharmazie, Jena, Germany, 5German Cancer Consortium (DKTK), Heidelberg, Germany, 6 German Cancer Research Center (DKFZ), Heidelberg, Germany, 7DKMS, German Bone Marrow Donor Center, Dresden, Germany 1

Introduction: Clofarabine (clo.) is a nucleoside analogue with activity in acute myeloid leukemia (AML). Liver toxicity, particularly a transient elevation of transaminases is a common side effect of this substance. Especially in the context of the allogeneic HSCT, liver toxicity is a major concern. We studied the correlation of pharmacokinetic parameters with the liver toxicity and antileukemic activity of clo. in patients (pts.) with relapsed or refractory (r/r) AML. Methods: We analyzed samples of pts. treated within the BRIDGE trial, a Phase II trial with clo. in combination with cytarabine as salvage therapy prior to hematopoietic stem cell transplantation (HSCT) in pts. with r/r AML. The pharmacokinetic parameters area under the curve (AUC) from 0–24 h after end of the infusion and the maximum plasma level (cmax) were analyzed and correlated with daily transaminase and bilirubin levels on days 0–15 after the first clo. infusion, with the maximum levels of transaminases and with response to salvage chemotherapy. Linear mixed models with random intercept and random slope and AUC per day exposure as influential variable were fitted in order to assess the impact of drug exposure on courses of transaminase and bilirubin levels. Multiple regression models were fitted to adjust for age, sex, clo. dose, baseline transaminase and treatment cycle. Results: The mean transaminase levels rose within 10 days after start of clofarabine therapy with a maximum of more than 3.5 times the upper limit of normal between day 5 and 7. Up to day 15 the levels declined to the baseline values in all pts. Results of the plasma levels showed large inter-individual differences in the pharmacokinetic parameters. A higher clo. AUC was significantly associated with liver toxicity reflected by a higher transaminase elevation (p = 0.02 for ALAT, adjusted for age, sex, Clo. dose, baseline ASAT and treatment cycle). No statistical significant correlation could be found between cmax and the liver toxicity parameters. Furthermore, no association between the response to salvage chemotherapy evaluated at day 15 and the AUC or cmax of clo. was observed. Conclusion: We were able to show that a higher individual clo. exposure is associated with more severe liver toxicity reflected by elevated liver enzymes without having an impact on anti-leukemic efficacy. Therefore, pharmacokinetic analysis might help to calibrate the optimal dosing of clo. for individual pts. Disclosure: Bozena Büttner: No conflict of interest disclosed. Johannes Schetelig: Expert Testimony: Sanofi (Genzyme)

Oncol Res Treat 2016;39(suppl 3):1–104

Medizinische Fakultät Carl Gustav Carus der TU Dresden, Medizinische Klinik und Poliklinik 1, Dresden, Germany, 2Deutsches Krebsforschungszentrum, Molekulare Hämatologie/Onkologie, Heidelberg, Germany, 3 Universitätsklinikum Münster, Medizinische Klinik A, Münster, Germany, 4J.-W.Goethe-Universität, Medizinische Klinik II, Frankfurt, Germany 1

Mutations of the key myeloid transcription factor CCAAT/enhancer binding protein alpha (C/EBPa) are found in 5–10% of patients with acute myeloid leukemia (AML). Two mutational clusters exist, in the aminoterminal transcription activation domains (TAD1 or 2) and in the basic leucine zipper domain (bZIP) located at the c-terminus of the protein. Biallelic mutations (biCEBPA) have been found to be associated with improved outcome and are now included as an independent entity in the WHO-classification. In contrast, monoallelic CEBPA-mutations (moCEBPA) do not appear to provide prognostic information. We analyzed the mutational spectrum of mono- and biallelic CEBPA-mutant AML patients to better understand potential differences in the biology of these groups. Patients and methods: Patients included were registered clinical protocols of the Study Alliance Leukemia (SAL)(AML96, AML2003 or AML60+, SORAML) or the SAL-register. Screening for CEBPA mutations was done using PCR and capillary electrophoresis. All CEBPA mutant samples were analyzed using next generation sequencing (Trusight Myeloid Panel, Illumina). Results: 239 patients with AML and CEBPA-mutations were identified. We restricted the analysis of coexisting mutations to 204 patients with normal karyotype (NK) or intermediate-2 mutations according to the ELN-classification. In this group, 104 patients presented with biCEBPA, whereas 100 had moCEBPA (57 bZIP, 43 TAD). Interestingly, these groups showed significantly differing patterns of co-mutations. In patients with biCEBPA, the most common alterations were seen in GATA2 (39%), TET2 (24%), WT1 (22%), and NRAS (15%). A similar distribution was found in bZIP-moCEBPA mutant patients (GATA2 (37%), TET2 (23%), WT1 (11%), and NRAS (14%)). In contrast, patients with isolated mutations in the TAD-domains had only 2% GATA2 mutations, predominant lesions were found in TET2 (42%), NPM1 and DNMT3A (35% each) and FLT3ITD (26%), a pattern reflecting the distribution seen in the general AML population with NK. An analysis comparing the effect of these alterations on outcome indicated that patients with GATA2 mutations had an improved EFS but no significant difference in the OS and CR rate, whereas TET2-mutations were associated with significantly worse OS and EFS. Conclusions: These results support functional differences between the mutational subgroups in CEBPA and reveal for the first time clinically relevant differences among the moCEBPA patient subgroup. Disclosure: Julia-Annabell Georgi: No conflict of interest disclosed. Christian Thiede: Employment or Leadership Position: AgenDix GmbH; Stock Ownership: AgenDix GmbH

Abstracts

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V182

Single cell genotyping and epigenotyping of acute myeloid leukemia Renz N.1, Niemöller C.1, Riba J.2, Bleul S.1, Metzeler K.H.3, Stosch J.M.1, Pfeifer D.1, Duyster J.1, Lübbert M.1, Zimmermann S.2, Claus R.1, Becker H.1 Medical Center – University of Freiburg, Department of Medicine I, Freiburg, Germany, 2University of Freiburg, Department of Microsystems Engineering – IMTEK, Freiburg, Germany, 3University of Munich, Department of Medicine II, Munich, Germany 1

Clonal genetic and epigenetic heterogeneity impacts outcome in acute myeloid leukemia (AML). Deciphering the clonal heterogeneity requires analyses in single cells. Here, we studied gene mutations and DNA methylation in AML bulk samples and single cells. In bulk samples, mutations were analyzed by targeted next generation-, Sanger- or pyrosequencing and DNA methylation by HumanMethylation450K array (Illumina). Single cells were deposited into microtiter plates by a Single-Cell Printer (SCP). Mutations in single cells were studied by PCR/Sanger sequencing directly on single cell DNA or after whole genome amplification (WGA); WGA success was assessed by a PCR on LINE1 transposons. DNA methylation in single cells was analyzed by methylation sensitive restriction enzyme digest (MSRE-D). Mutations in KIT (p.N822K; variant allele frequency [VAF] 84%) and TP53 (p.R248Q; VAF 100%) were identified in the AML cell line Kasumi-1. Of the 450,000 DNA methylation sites analyzed in Kasumi-1, we selected 4 loci in DNMT3A with 3%, 70%, 77% and 96%, respectively, methylation level for further analyses. We printed 33 single Kasumi-1 cells and subjected them to WGA. Median DNA yield per cell was 14µg; the LINE1 PCR was positive in all cells. The KIT mutation was confirmed in 30, the TP53 mutation in 25 cells; PCR in the remaining cells failed. All cells with results for both mutation sites harbored both the KIT and TP53 mutation. The DNMT3A site with 3% methylation level in the bulk expectedly was unmethylated in 7 cells analyzed by MSRE-D. Preliminary analysis of both DNMT3A methylation and TP53 within the same cell was successful in 2 cells. We also studied single cells of a patient AML that in bulk sequencing only had the G-allele detectable in the TP53 polymorphism rs1042522 (p.P72R) and had a subclonal monosomy 17 (hosting TP53). Among 23 single cells, median WGA DNA yield was 16µg; the LINE1 PCR was positive in all cells. The TP53 PCR yielded a product in 20 cells. In 5 cells we detected the C-allele which was not found in bulk sequencing, suggesting its subclonal loss due to monosomy 17. In conclusion, we confirmed the co-occurrence of mutations and verified the DNA methylation status in single Kasumi-1 cells. Single cell genotyping also confirmed subclonal genetic changes in an AML patient. Evaluation of mutation and methylation status in the same cell allows unique insights into clonal architecture; the SCP proved to be an efficient tool for single cell isolation.

duce expression of Wnt downstream target genes. Aberrant expression of Lef1 was shown to perturb normal hematopoietic stem cell (HSC) and progenitor function, induces AML in the murine bone marrow transplantation model and is a novel prognostic independent factor in patients with normal karyotype AML (Petropoulos et al. JEM 2008; Metzeler et al., Blood 2012). Furthermore we could show that Lef1 is essential for LT-HSC function (Edmaier et al. Leukemia, 2014). To dissect the role of the Lef1 isoform lacking the β-catenin binding domain at the N-terminus (Lef1ΔN), we retrovirally engineered primary murine bone marrow cells to express the Lef1ΔN in comparison to the full-length Lef1 (Lef1WT) and the empty retroviral YFP control. Loss of binding to β-catenin was validated by Co-IP for Lef1ΔN. The mutant did not impact Lef1 activity at the level of the short-term repopulating stem cells (CFU-S) but reduced the CRU frequency substantially (1:597.197(Lef1WT) versus 1:1.675.238 (Lef1ΔN), respectively, and 1: 1.233.152 for the EGFP control), indicating the necessity of Lef1 to collaborate with β-catenin at the level of the LTHSCs. ChIP-Seq in hematopoietic murine cells revealed unique binding sites for Lef1ΔN compared to Lef1WT with binding of the Lef1ΔN to the promoter of Wnt5a, which is known to regulate HSC proliferation and repopulation (Povinelli et al., Stem cell 2014). To further understand the relevance of expression of the different LEF1 isoforms in patients with AML, over 100 normal karyotype AML cases were analysed by qRT-PCR, showing a dominant expression of the long isoform of LEF1 in the leukemic bulk population. Dependence of leukemic growth on binding of LEF1 to β-catenin was further indicated by a high sensitivity of AML cell lines to the compounds Cercosporin and Calphostin C, which were shown to block LEF1-β-catenin interaction. Our data clearly indicate that loss of the β-catenin binding site creates a ‘neomorphic’ isoform with distinct biological and DNA binding properties. Differential expression of LEF1 isoforms in normal versus leukemic stem and progenitor cells might open the way to target leukemic cells by blocking LEF1-β-catenin crosstalk. Disclosure: No conflict of interest disclosed.

Posterdiskussion

Akute myeloische Leukämie P186

Non-psychoactive cannabidiol exerts proapoptotic activity in acute leukemia cells in vitro and ex vivo Kampa-Schittenhelm K.1, Häusser L.1, Kanz L.1, Schittenhelm M.1 Universitätsklinikum Tübingen, Medizinische Klinik II, Tübingen, Germany

The Wnt pathway is essential for maintenance, activation and proliferation of normal hematopoietic stem cells. The lymphoid enhancer factor 1 (Lef1) is a member of the Lef/T-cell-factor family of transcription factors that directly interacts with β-catenin in transcriptional complexes to in-

We recently demonstrated that (−)-Δ9-Tetrahydrocannabinol (THC) has potent antileukemic activity in acute myeloid and lymphoblastic leukemia (AML/ALL) in vitro and in vivo. However, clinically effective doses may not be achieved due to compliance and side effect issues – especially with regard to the unwanted psychotropic effects of THC. Cannabidiol (CBD) is a natural cannabinoid derivative with minimal psychotropic activity. It is part of a combination compound approved as Sativex for the treatment of spasms in multiple sclerosis. We herein provide evidence that CBD is even more potent with regard to the proapoptotic antileukemic activity compared to THC in several acute leukemia cell models in vitro as well as in ex vivo native blasts. The AML cell lines MOLM13 and MOLM14 and the T-lymphoblastic Jurkat cell line as well as native blasts from patients were comparatively treated with CBD and THC in dose-dilution series and assessed for induction of apoptosis (Annexin V-based flow cytometry assay) and anti-proliferative effects (XTT). MOLM13/14 and Jurkat cells displayed potent induction of apoptosis upon exposure to CBD with IC50s that were significantly lower compared to THC (5–7 microM vs.10–15 microM). This observation was confirmed treating native leukemia blasts of 10 patients with AML (n = 7) or ALL (n = 3) with CBD and THC ex vivo. Knockout (ko) of the cannabinoid

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

Disclosure: No conflict of interest disclosed. V183

The Lef1 isoform, lacking the β-catenin binding domain, is not acting as a dominant negative Lef1 variant, but is a hematopoietic active protein with unique DNA binding properties Feder K.1, Edmaier K.1, Eshraghi P.1, Vegi N.1, Mulaw M.1, Ihme S.1, Spiekermann K.2, Metzeler K.2, Hiddemann W.2, Döhner K.3, Döhner H.3, Feuring-Buske M.3, Buske C.1 Institute for Experimental Cancer Research, University Hospital Ulm, Ulm, Germany, 2Department of Internal Medicine III, University Hospital Munich, Grosshadern, Munich, Germany, 3Department of Internal Medicine III, University Hospital Ulm, Ulm, Germany 1

CONTENTS AUTHOR INDEX

receptors CB1 and CB2 in different leukemia cell models was performed using an shRNA approach. Pathway signaling analyses via the different cannabinoid receptors is ongoing. First data suggests apoptosis induction via both receptors, since CB1 as well as CB2 ko cells demonstrate attenuated apoptosis for both substances compared to the empty vector controls. Besides a pro-apoptotic effect on leukemia blasts we have recently demonstrated that THC is capable of overriding the differentiation block in vivo resulting in loss of CD34 expression and an increase of CD11c, CD14 and/ or CD15 in two patients treated with dronabinol for best supportive care – resulting in a maturing monocytic population of leukemic origin. These findings were reproducible in in vitro cell models. So far, similar effects were not seen for CBD, but shedding light into the mechanisms of differentiation is topic of our ongoing research. Our findings provide a strong rationale to clinically explore CBD as an agent with remarkable antileukemic efficacy. Disclosure: No conflict of interest disclosed. P187

Proof of clinical activity of Dasatinib in a patient with mutantKIT D816V positive core binding factor leukemia (CBF AML) Schittenhelm M.1, Bonzheim I.2, Witte K.1, Soekler M.1, Kanz L.1, KampaSchittenhelm K.1 Universitätsklinikum Tübingen, Medizinische Klinik II, Tübingen, Germany, Universitätsklinikum Tübingen, Institut für Pathologie, Tübingen, Germany

1 2

Activating KIT D816V mutations are frequently found in CBF AML and are predictive for an unfavorable outcome. Dasatinib is a potent inhibitor of wildtype and mutant-KIT isoforms – including D816V. However, clinical efficacy as monotherapy is in question. In this study, we proof antileukemic efficacy via overriding the differentiation block in a patient with mutant-KIT D816V CBF AML. Clinical activity of dasatinib was monitored using (1) Western immunoblotting for target-specificity (2) a plasma-inhibitory assay (PIA) using serum of this patient and our well established KIT D816V-dependent Ba/ F3 model cell line to demonstrate clinically active concentrations (3) cytomorphology and immunophenotyping to analyze differentiation effects (4) a cell sort to isolate the maturing population and screen for presence of KIT D816V in order to proof leukemia origin. This 77 year old unfit male patient with relapsing disease (WBC 13 000/ microL, blasts 59%) and a high KIT D816V mutation burden was put on dasatinib 70 mg BID. Target-specificity was demonstrated via dephosphorylation (i.e. inactivation) of KIT on day 2 and 4 and PIA revealed clinically active doses of dasatinib (49% apoptotic cells after 48hrs). Intermittent cytoreduction with hydroxyurea was necessary due to leukocytosis, which did not allow to definitely proof proapoptotic efficacy of dasatinib in vivo. Intriguingly however, an increasing monocytic population (up to 80%) was noted – going along with disappearance of morphologic blasts and rise of neutrophils (~20%). Immunophenotyping argued for maturation of the leukemia clone, shifting from a CD14+/CD34+ positive phenotype to CD34 negativity. However, due to suspected pneumonia and questionable disease recurrence, therapy was halted. High doses of dexamethasone quickly improved the symptoms strengthening the differential diagnosis of differentiation syndrome. Cytoreduction with cytarabine and 6-thioguanin resolved leukocytosis and dasatinib was readministered with 100 mg/d. Again, signs of maturation were documented. Release of differentiation block was confirmed ex vivo demonstrating loss of CD34 upon exposure to dasatinib. Ultimately, sanger sequencing of the maturing population provided molecular proof of the leukemic origin with presence of KIT D816V in the suspected CD14+/CD34- population. To conclude, dasatinib has clinical activity in mutant-KIT CBF AML via effective release of the differentiation block – providing a novel therapeutic approach. Disclosure: No conflict of interest disclosed.

Oncol Res Treat 2016;39(suppl 3):1–104

P188

Oncometabolite 2-hydroxyglutarate impacts metabolism but not function of T cells Böttcher M.1, Renner K.2, Mentz K.1, Mackensen A.1, Kreutz M.2, Mougiakakos D.1 University Hospital Erlangen, Department of Internal Medicine 5, Hematology and Oncology, Erlangen, Germany, 2University Hospital Regensburg, Department of Internal Medicine III, Regensburg, Germany 1

Elevated levels of the oncometabolite 2-hydroxyglutarate (2HG) have been described in different tumor entities such as glioma, breast cancer, and AML both in serum and in the tumor. It was shown that 2HG can be generated via conversion of α-ketoglutarate by mutated isocitrate dehydrogenases (IDH), a process taking also place with non-mutated IDH but at a much lower level, or in a myc-driven manner via glutaminolysis. However, current research has thus far only focused on 2HG-mediated intrinsic effects on the tumor itself, especially its role as a competitive inhibitor of α-ketoglutarate resulting in epigenetic changes as well as metabolic deregulation. Moreover, results are often converse. Here, we show for the first time the effects of D-2HG (the most common enantiomer in this context) on activated T cells. Upon treatment with D2HG T cell respiration (basal and maximal), as well as respiration-related ATP production were enhanced while glycolysis was decreased. Additionally, glucose uptake, mTOR phosphorylation, as well as the content of α-ketoglutarate and citrate were elevated while lactate production was reduced. On the other hand, no influences on proliferation, survival, oxidative and suppressive capacity, T cell activation and memory formation, as well as mitochondrial biomass and membrane potential could be shown. Taken together, tumor-derived D-2HG affects T cell metabolism by enhancing mitochondrial respiration and decreasing glycolysis. Considering the elevation of tricarboxylic acid cycle intermediates D-2HG might fuel respiration by anaplerosis. However, the underlying mechanism and the possible impact on T cell activation and function remains to be elucidated. Ultimately, long-term effects of D-2HG as found in tumor patients should be explored as well. Disclosure: No conflict of interest disclosed. P189

Assessment of prognosis and minimal residual disease in acute myeloid leukemia based on the use of NPM1 gene mutations in patients of Gomel Region, Belarus Kozich Z.1, Silin A.1, Martinkov V.1, Tropashko I.1 Das Republikanische Centrum für Strahlungsmedizin und Humanökologie, Hämatologie, Gomel, Belarus 1

Introduction: Considering the heterogeneity of acute myeloid leukemia (AML), the intensity of chemotherapy depends on the specific number and combination of morphological and molecular genetic markers. The additional use of specific leukemic markers (such as mutations in NPM1 gene) will increase the effectiveness of the therapy through the timely identification of them in the period of remission as an early sign of recurrence, as well as evaluate the prognosis of the disease. Methods: The material for the study served as samples of venous blood and bone marrow of 111 patients with AML, who were treated in the period 2009–2014 in hematology department of SI “RRCRM & HE”, Gomel. To identify mutations in NPM1 gene there was used PCR method (PCR) with specific primers followed by electrophoretic detection. Results: As a result of molecular-genetic analysis it was found that 26 of 111 analyzed patients (23.4%) were carriers of NPM1 mutations. In 10 cases, the mutation was identified together with FLT3-ITD mutation. The highest frequency of mutations was observed for M1 variant (28.6%). In groups of patients with M2, M3 and M4 variants NPM1 mutations were identified in 24.0%, 16.0% and 23.5% respectively. In patients with M0 variant NPM1 mutations were absent. In the analyzed group of patients in the age of less than 60 years at the moment of diagnosis of AML the presence of NPM1 mutations was statistically significant

Abstracts

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adverse prognostic factor especially in combination with FLT3-ITD mutation. In addition survival rate in patients with leukocytosis in blood of more than 20 *10^9 /L and the blood Hb less than 90 g/L was significantly lower in relation to patients with normal levels of Hb and indicators of leukocytes of less than 20 *10^9 L. A mutation in NPM1 gene, preserved after the first course of chemotherapy was associated with a high risk of relapse after three years of follow-up treatment and low overall survival. Although this mutation is connected with preleukemic clone, in 2 patients it was determined throughout remission after chemotherapy. During relapse, the mutation in NPM1 gene was identified in all patients that initially had this mutation. Conclusion: Determination of mutations in NPM1 gene can supplement the prognostic information of other risk factors that will help to improve the survival of patients with AML as well as to increase the effectiveness of treatment when using NPM1 mutation as a molecular marker of relapse. Disclosure: No conflict of interest disclosed. P190

Differential expression of Pri-miR-320a impacts on outcome in acute myeloid leukemia patients undergoing nonmyeloablative allogenic stem cell transplantation Gaber T.1, Bill M.1, Jentzsch M.1, Schubert K.1, Weidner H.1, Grimm J.1, Schulz J.1, Kloss L.1, Schmalbrock L.1, Bonifacio L.1, Wildenberger K.1, Pönisch W.1, Vucinic V.1, Franke G.-N.1, Lange T.1, Cross M.1, Behre G.1, Niederwieser D.1, Schwind S.1 Universitätsklinik Leipzig, Hämatologie und internistische Onkologie, Leipzig, Germany 1

MicroRNA (miR) expression has been shown to be altered in AML & miR-based therapies are entering clinical trials. MiR-320a maps to chromosome 8p21.3 & its high expression has been associated with improved outcomes in some solid cancers. In AML miR-320 inhibits cell proliferation, likely by targeting the transferrin receptor 1. The objective of this study was to investigate whether a differential expression of miR-320a also impacts on outcome in AML pts. We analyzed 121 AML patients (pts) (median age at diagnosis 63 years [y]; range 37–75 y) who were treated with cytarabine-based chemotherapy & received non-myeloablative allogeneic stem cell transplantation (NMASCT, Fludarabine 3×30mg & 2 Gy total body irradiation) as consolidation therapy at our institution. At diagnosis, pts were characterized for presence of FLT3-ITD, CEBPA, DNMT3A, IDH1, IDH2 & NPM1 mutations & EVI1 expression. European LeukemiaNet (ELN) genetic classification was: 26% favorable, 26% intermediate I, 21% intermediate II & 26% adverse. Pretreatment pri-miR-320a expression in bone marrow (BM) was analysed by reverse transcription polymerase chain reaction & normalized to a housekeeping gene (18S). The median normalized gene expression defined high & low pri-miR-320a expressers. Median follow-up was 4.3y for pts alive. High pri-miR-320a expression associated with younger age (P = 0.02), lower platelet count (P = 0.04) & by trend lower BM blast count (P = 0.08) at diagnosis. High pri-miR-320a expressers more frequently were IDH2 (P = 0.04) & DNMT3A (P = 0.05) mutated & were less likely to be EVI1 positive (P = 0.02). High pri-miR-320a expressers had significantly longer overall survival (OS; P = 0.04; Figure 1a) & event-free survival (EFS; P = 0.02, Figure 1b). In multivariate analysis, pri-miR-320a expression kept its prognostic impact on EFS (Hazard Ratio 0.5 [95% Confidence Interval 0.3–0.9], P = 0.01).

Abstracts

Fig. 1. miR320.

We showed for the first time that pretreatment pri-miR-320a expression levels impact on outcome in AML pts. Increasing miR-320a expression prior to AML treatment, e.g. by miR-replacement therapies, might improve outcomes of AML pts, especially ones with low miR-320 expression. Disclosure: No conflict of interest disclosed. P191

In AML with normal karyotype lacking mutations in NPM1, CEBPA, RUNX1 and MLL partial tandem duplication array CGH and fluorescence in situ hybridization can add prognostic information Haferlach C.1, Kern W.1, Zieschang K.1, Zenger M.1, Perglerova K.2, Haferlach T.1 MLL Münchner Leukämielabor GmbH, München, Germany, 2MLL 2, Paříkova, Czech Republic 1

Introduction: A subset of AML with normal karyotype lacking mutations in NPM1, CEBPA, RUNX1 and a partial tandem duplication within the MLL gene (MLL-PTD) is still poorly characterized. Array CGH and fluorescence in situ hybridization are able to detect abnormalities which are undetectable by chromosome banding analysis either due to a higher resolution, ability to detect copy neutral loss of heterozygosity (CN-LOH) and independence of in vitro proliferation. Patients and methods: For 1473 AML cases with normal karyotype complete data on mutation status of NPM1, CEBPA, RUNX1 and MLL-PTD was available. Of these 303 cases (21%) did not carry any of these mutations. Out of these 159 cases with de novo AML (median age: 68 years (range: 19–93)) were selected based on the availability of material for array CGH (SurePrint G3 ISCA CGH+SNP Microarray, Agilent) and FISH screening with probes for MLL, RUNX1, CBFB, NUP98, MECOM/EVI1, NPM1, ETV6 and DEK-NUP214 (MetaSystems, Abbott). Results: In total in 67 of 159 patients (42%) abnormalities were identified by FISH and/or array CGH. In detail, 12 balanced rearrangements were detected by FISH screening involving NUP98 (n = 7), MLL (n = 2) and MECOM, RUNX1 and CBFB (one each). In addition, 27 gains, 42 losses and 41 CN-LOH were observed in 58 (37%) patients. Recurrent gains affected regions 6q23.3q23.3 (n = 2) and 8q24.21q24.21 (n = 2) while recurrent losses were found for the regions 21q22.12q22.12 encompassing the RUNX1 gene (n = 5), 5q31.2q31.2 including i.a. EGR1 and CTNNA1 (n = 3), 2q34q34 including i.a. IKZF2 (n = 2), 7q22.1q22.1 encompassing i.a. CUX1 (n = 2), and Yq11.223q12 (n = 2). Recurrent CN-LOH were observed on chromosomes 11q (n = 10), 2p (n = 5), 4q (n = 4), 21q (n = 3), 1p (n = 2), 17q (n = 2) and 18q (n = 2). Survival analysis was performed for 90 intensively treated patients. Overall survival (OS) and event-free survival (EFS) were significantly shorter in patients with abnormalities detected by FISH and/or array CGH compared to those without (median OS: 19 vs 49 months, p = 0.027; median EFS: 9 vs 21 months, p = 0.016). Conclusions: 42% of AML with normal karyotype lacking a disease defining molecular mutation harbor balanced rearrangements, copy number gains or losses as well as CN-LOH. The presence of these abnormalities

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has a negative impact on survival demonstrating that FISH and array CGH can add prognostic information in AML with normal karyotype. Disclosure: No conflict of interest disclosed. P192

Three cases of cyclic neutropenia with acquired CSF3R mutations, one developing AML Klimiankou M.1, Zeidler C.2, Mellor-Heineke S.2, Reinel E.1, Kandabarau S.1, Uenalan M.3, Kanz L.4, Welte K.5, Skokowa J.1 University Hospital Tübingen, Department of Hematology, Oncology, Immunology, Rheumatology and Pulmonology, Division of Translational Oncology, Tübingen, Germany, 2Hannover Medical School, Severe Chronic Neutropenia International Registry (SCNIR), Hannover, Germany, 3Hannover Medical School, Department of Experimental Hematology, Hannover, Germany, 4 University Hospital Tübingen, Department of Hematology, Oncology, Immunology, Rheumatology and Pulmonology, Tübingen, Germany, 5University Hospital Tübingen, Dept of Pediatrics, Tübingen, Germany 1

Introduction: We recently reported a 17-year-old female with cyclic neutropenia (CyN) harboring ELANE mutations presenting with cycling hematopoiesis involving neutrophils, platelets and reticulocytes who developed AML (FAB M2) (presented at ASH 2015, Blood 126 (23) abstract 885). At the time of AML diagnosis, all CSF3R-expressing BM leukemic blasts were positive for the p.Gln741X mutation. We tested the patient’s BM MNCs, obtained at the time of leukemia development, for RUNX1 mutations, finding that the RUNX1 mutation p.Asp171Asn was present at an allele frequency of 10%. So far, CSF3R mutations have been reported in congenital neutropenia patients (CN) only but never in patients with CyN. Next, we aimed to determine whether other CyN patients harbor CSF3R mutations. Methods: We performed deep sequencing of CSF3R critical region in 19 additional CyN patients. Results: Out of these 19 patients we identified two additional patients harboring acquired CSF3R mutations. One CyN patient aged 15.4 years and her sister inherited the ELANE mutation from their father. Time-course analysis of the CSF3R mutations in this patient showed that at the age of 13 years frequency of the CSF3R p.Gln749X allele in her BM MNCs sample reached 2.6%. After additional 1.5 and 2.4 years, respectively, the mutant allele frequency increased to 9% and 8%. A third CyN patient, a 7 year old girl, harboring spontaneous ELANE mutations revealing typical cycling of neutrophils, platelets, and monocytes has been treated with G-CSF at a dose of 4.5 µg/kg/day. She has been recently detected with 30% of frequency of CSF3R p.Gln739X mutant allele. The latter 2 CyN patients have not developed AML or myelodysplastic syndrome (MDS). Conclusion: Taken together, these findings suggest that CyN patients with typical CyN-associated ELANE mutations may also acquire CSF3R mutations and are therefore at risk for leukemic development. Long-term data of the Severe Chronic Neutropenia International Registry (SCNIR) have shown that the risk of acquisition of CSF3R mutations and myeloid transformation is very low but not absent in patients with CyN compared to patients with CN. This new knowledge is important for prognostic counseling and long-term management of ELANE-CyN patients. Disclosure: No conflict of interest disclosed. P193

Idiopathic intracranial hypertension resulting from ATRA therapy in acute promyelocytic leukemia: an important adverse effect Teichmann L.L.1, Thieltges F.2, Brossart P.1, Mayer K.1 Universitätsklinikum Bonn, Medizinische Klinik III, Bonn, Germany, Universitätsklinikum Bonn, Augenklinik, Bonn, Germany

1 2

Introduction: All-trans retinoic acid (ATRA) is highly effective in inducing remission in patients with acute promyelocytic leukemia (APL) by causing the differentiation of abnormal promyelocytes to mature neutro-

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phils. Retinoids and vitamin A, however, are strongly associated with the development of idiopathic intracranial hypertension (IIH), a disorder of elevated intracranial pressure with no established pathogenesis. Case: A 21-year old woman, who received idarubicin plus ATRA as induction therapy for high-risk APL, presented with refractory headache and nausea accompanied by intermittent brief graying-out of vision. Bilateral enlargement of the blind spot was noted in the visual field test. Funduscopic evaluation revealed bilateral swelling of the optic disc with obscuration of major blood vessels and retinal hemorrhages. These findings were suggestive of papilledema due to raised intracranial pressure. The presence of papilledema was confirmed by optical coherence tomography. Magnetic resonance imaging of the brain showed no secondary causes of increased intracranial pressure such as intracranial bleeding or cerebral venous sinus thrombosis. Thus, the patient was diagnosed with IIH. Lumbar puncture opening pressure was not determined due to thrombocytopenia and a prolonged prothrombin time. ATRA was immediately discontinued and the patient was treated with the carbonic anhydrase inhibitor acetazolamide. A rapid clinical improvement was seen and swelling of the optical nerves decreased. After symptoms resolved ATRA was reinitiated at a 50% dose with concomitant prophylactic acetazolamide, which was initially well tolerated. Yet, during the 1st consolidation cycle IIH recurred with sixth nerve palsy of the left eye. Symptoms were successfully treated by discontinuing ATRA and administering topiramate, an anticonvulsant that also blocks carbonic anhydrase, in addition to acetazolamide. Subsequently, the patient received the 2nd and 3rd consolidation cycle including ATRA at reduced dose without any major complications. Conclusion: The diagnosis of IIH should be kept in mind in patients with APL. Owing to the use of ATRA its incidence is expected to be much higher in this group than in the general population. Female gender and obesity are considered additional risk factors for IIH. Untreated IIH can lead to retinal atrophy and permanent visual loss. After resolution of symptoms reinitiation of ATRA is possible when the patient is closely monitored. Disclosure: No conflict of interest disclosed. P194

The prognostic impact of tetraploidy in patients with Acute Myeloid Leukemia Mohr B.1, Röllig C.2, Kramer M.2, Oelschlägel U.2, Thiede C.2, Jost E.3, Schetelig J.2, Middeke M.2, Brümmendorf T.H.3, Serve H.4, Berdel W.E.5, Ehninger G.2, Bornhäuser M.2, Stölzel F.2, SAL Universitätsklinikum Dresden, Medizinische Klinik und Poliklinik 1, Dresden, Germany, 2Universitätsklinikum Dresden, Dresden, Germany, 3Uniklinik RWTH Aachen, Aachen, Germany, 4Universitätsklinikum Frankfurt, Frankfurt, Germany, 5 Universitätsklinikum Münster, Münster, Germany 1

Introduction: Karyotypes of AML blasts reveal different disturbances of chromosomal integrity such as structural aberrations and numerical changes of whole chromosomes. Most of these recurring aberrations have been integrated in prognostication systems. Gains of whole chromosomes often appear together with structural aberrations and could be later events during disease progression whereby distinct structural aberrations rather function as earlier lesions with a strong impact on prognosis. Patients with trisomy 8 fare well in the presence of translocation t(8;21) whereas patients presenting with a sole trisomy 8 do not have a favorable prognosis (Schaich M 2007). In addition, a sole trisomy 13 was shown to be a marker conferring an adverse prognosis in AML (Herold T 2014) as well as several trisomies resulting in a pure hyperdiploid karyotype revealed an adverse prognosis, too (Stölzel F 2016). A rare but recurring phenomenon in AML is the tetraploid karyotype showing not only the doubling of several chromosomes but rather the doubling of the whole diploid karyotype. Therefore, the aim of this study was to investigate the prognostic impact of tetraploid karyotypes in AML. Methods: The AML databases of the German SAL group were examined for tetraploid cases. The respective chromosome analyses were performed at diagnosis. The probability for overall survival (OS) was estimated according to the Kaplan-Meier method.

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Results: Between 1996 and 2015 n = 17 patients, median age 55 years, fulfilled the criterion of tetraploidy (median OS: 18.7 months, 95%-CI 6.1−Inf). Taking into consideration additional aberrations two groups of patients were defined: (1) pure tetraploidy and ≤2 additional aberrations (n = 8) with a CR rate of 75% and a median OS of 26.2 months (95%-CI 18.7−Inf). (2) tetraploidy and ≥3 additional aberrations and at least one additional adverse-risk aberration (n = 9) with a CR rate of 67% and a median OS of 9.6 months (95%-CI 5.5−Inf). By comparison, in our SAL trials AML patients with a normal karyotype have a median OS of 24.9 months (95%-CI 22−27.9) and patients with adverse cytogenetics a median OS of 7.9 months (95%-CI 7.4−8.6). Conclusions: However, taking into consideration the low patient numbers, tetraploid AML patients without additional adverse risk aberrations have an OS comparable to patients with a normal karyotype. Tetraploid AML patients with additional adverse risk aberrations have an inferior OS comparable to adverse risk patients.

Tab. 1. Baseline Characteristics of Ongoing Patients

Disclosure: No conflict of interest disclosed. P195

Characterization of patients with relapsed or refractory AML in continued follow-up after treatment with vosaroxin/ cytarabine vs placebo/cytarabine in the valor trial Horst H.-A.1, Ravandi F.2, Ritchie E.K.3, Lancet J.E.4, Craig M.D.5, Pigneux A.6, Maertens J.7, Derigs H.-G.8, Heuser M.9, Wei A.10, Hogge D.11, Clark R.12, Ward R.13, Craig A.R.13, Stuart R.K.14 Klinik fuer Innere Medizin II, University Hospital Schleswig-Holstein, Kiel, Germany, 2University of Texas MD Anderson Cancer Center, Houston, United States, 3Weill Cornell Medical Center, New York, United States, 4Moffitt Cancer Center, Tampa, United States, 5West Virginia University, Morgantown, United States, 6Université de Bordeaux, Bordeaux, France, 7Universitair Ziekenhuis, Leuven, Belgium, 8Klinikum Frankfurt Main, Frankfurt, Germany, 9Hannover Medical School, Hannover, Germany, 10The Alfred Hospital and Monash University, Melbourne, Australia, 11Vancouver General Hospital, Vancouver, Canada, 12Royal Liverpool University Hospital, Liverpool, United Kingdom, 13 Sunesis Pharmaceuticals, Inc., South San Francisco, United States, 14Hollings Cancer Center, Medical University of South Carolina, Charleston, United States 1

Introduction: Patients with relapsed/refractory (R/R) AML have a median overall survival (OS) less than 1 year. In the phase 3 VALOR trial, vosaroxin/cytarabine prolonged median OS in patients with R/R AML by 1.4 mo vs placebo/cytarabine (7.5 vs 6.1 mo; HR = 0.87 [95% CI 0.73–1.02]; p = 0.061). Of 711 enrolled patients, 134 (19%) were alive in follow-up at the primary analysis. Here, we characterize patients in continued follow-up. Methods: In VALOR, patients with R/R AML were randomized 1:1 to receive cytarabine (1 g/m2 IV over 2 h, d 1–5) plus either vosaroxin (90 mg/ m2 IV over 10 min d 1, 4; 70 mg/m2 in subsequent cycles) or placebo. After the primary analysis, ongoing patients were followed for survival. Results: As of Jan 22, 2016, 83 patients (12%) were alive in follow-up: 46/356 (13%) in the vosaroxin/cytarabine arm and 37/355 (10%) in the placebo/cytarabine arm. Median follow-up in these patients was 40 mo (range 28–60). Patient characteristics are presented (Table); a higher proportion of patients were ≥ 60 years in the vosaroxin/cytarabine arm (50% vs 27% with placebo/cytarabine). Most achieved complete remission (CR) on study (70% with vosaroxin/cytarabine; 51% with placebo/cytarabine); over half maintained CR at database lock (59% with vosaroxin/cytarabine; 49% with placebo/cytarabine). Nearly all had subsequent therapy (93% with vosaroxin/cytarabine; 100% with placebo/cytarabine). Most patients on vosaroxin/cytarabine (85%) and all patients on placebo/cytarabine received posttreatment stem cell transplantation (SCT). Seven patients in the vosaroxin/cytarabine arm did not undergo SCT; all were ≥ 60 years of age. Median follow-up in these 7 patients was 33 mo (range 31–48).

Abstracts

Conclusions: A small proportion of patients with R/R AML continue to be followed for survival in VALOR. Typically, these patients achieved CR followed by SCT; however, some patients ≥ 60 years treated with vosaroxin/cytarabine achieved long-term survival without SCT. Disclosure: Heinz-August Horst: Advisory Role: Amgen, AbbVie, BMS; Financing of Scientific Research: Amgen, AbbVie; Expert Testimony: Amgen, Boehringer Ingelheim; Other Financial Relationships: MSD, BMS, Celgene Robert Stuart: Advisory Role: Sunesis; Financing of Scientific Research: Sunesis; Expert Testimony: Sunesis P196

Evaluation of tyrosine kinase inhibitor treatment in patients with FLT3-ITD positive acute myeloid leukemia Fleischmann M.1, Schrenk K.G.1, Schnetzke U.1, Hilgendorf I.1, Hochhaus A.1, Scholl S.1 Klinik für Innere Medizin II, Universitätsklinikum Jena, Hämatologie/ internistische Onkologie, Jena, Germany 1

Introduction: Activating mutations of the receptor tyrosine kinase FLT3 (fms-related tyrosine kinase 3) represent the most frequent molecular aberration in acute myeloid leukemia (AML) and predominantly comprise FLT3 internal tandem duplications (FLT3-ITD) that are associated with a worse outcome. Targeted therapy with tyrosine kinase inhibitors (TKI) addressing FLT3-ITD has been introduced into the treatment of relapsed or refractory AML. Methods: We retrospectively analysed the efficacy of TKI treatment in 15 patients (median age 54 years, range 21–74, 10 female) with relapse of FLT3-ITD positive AML. All patients underwent sorafenib treatment while 3 of 15 patients also received 2nd line therapy with ponatinib. Nine of 15 patients (60%) were characterised by AML relapse after allogeneic stem cell transplantation. Response to TKI treatment was evaluated according to the criteria published by Metzelder et al. (Blood 2009). Furthermore, TKI-related toxicity was analysed. Results: Hematologic response (HR) was observed in 7 of 15 patients (47%) while only two patients (13%) achieved bone marrow response (BMR) during treatment with sorafenib. In contrast, there was no patient who achieved a complete (CR) or even complete molecular remission (CMR). The median duration of sorafenib therapy was 73 days (11–213). Importantly, in the subgroup of patients with AML relapse after allogeneic stem cell transplantation only 3 of 9 patients (33%) responded to sorafenib

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treatment (all with HR only). In addition, all patients who received ponatinib as 2nd line treatment had no response. TKI-induced toxicity was moderate and only one patient developed grade 3 toxicity (rash) related to sorafenib. Conclusions: Treatment of relapsed FLT3-ITD positive AML remains a huge challenge and in most cases treatment with TKIs like sorafenib is not sufficient to control this disease. Especially in patients with AML relapse following allogeneic stem cell transplantation the results of TKI treatment are dissatisfying. Results of prospective clinical trials investigating 2nd generation TKIs (e.g. quizartinib, AC220) or concepts combining epigenetic approaches with FLT3-ITD directed treatment are needed. Disclosure: Maximilian Fleischmann: No conflict of interest disclosed. Sebastian Scholl: Expert Testimony: Förderprogramm Infektiologie der Firma Gilead P197

Patients with acute myeloid leukemia admitted to intensive care units: Scores predicting outcome and post-ICU survival Pohlen M.1, Braess J.2, Thudium J.3, Schmid C.4, Kochanek M.5, Kreuzer K.A.5, Görlich D.6, Gerth U.7, Rhode C.8, Müller-Tidow C.8, Stelljes M.1, Büchner T.1, Schlimok G.4, Hallek M.5, Waltenberger J.9, Hiddemann W.3, Berdel W.E.1, Heilmeier B.2, Krug U.10 Universitätsklinikum Münster, Medizinische Klinik A – Hämatologie und Onkologie, Münster, Germany, 2Krankenhaus der Barmherzigen Brüder, Onkologie und Hämatologie, Regensburg, Germany, 3Universitätsklinikum Großhadern, Hämatologie und Onkologie, München, Germany, 4Klinikum Augsburg, Medizinische Klinik II, Augsburg, Germany, 5Universitätsklinikum Köln, Medizinische Klinik I, Köln, Germany, 6Universitätsklinikum Münster, Institut für Biometrie und Statistik, Münster, Germany, 7Universitätsklinikum Münster, Medizinische Klinik D – Innere Medizin, Nephrologie und Rheumatologie, Münster, Germany, 8Universitätsklinikum Halle-Wittenberg, Medizinische Klinik IV, Halle/Saale, Germany, 9Universitätsklinikum Münster, Department für Kardiologie, Münster, Germany, 10Klinikum Leverkusen, Medizinische Klinik 3, Leverkusen, Germany 1

Introduction: Despite encouraging survival rates of ICU survivors compared to non-ICU patients, assumed high mortality represents a major reason for the widespread hesitation to refer AML patients for treatment in the ICU. This retrospective, multicenter study aimed to reveal risk predictors for mortality in the ICU as well as survival after ICU discharge in patients with AML requiring treatment in the ICU. Methods: The AML in ICU score was established in a cohort of 187 adults with AML admitted to the ICU at the University Hospital of Muenster between 11/2004 and 09/2011. Validation was performed on a cohort of 264 patients with AML admitted to the ICU at the University Hospital of Grosshadern in Munich, the University Hospital of Cologne and the Municipal Hospital of Augsburg (all located in Germany) between 01/2004 and 02/2010. Results: Multivariate analysis of training cohort revealed the following as independent prognostic factors for death in the ICU: arterial oxygen partial pressure below 72 mmHg, active AML and systemic inflammatory response syndrome upon ICU admission, and need for hemodialysis and mechanical ventilation. Based on these variables, we developed an ICU mortality score and validated the score in the validation cohort. Compared with the Simplified Acute Physiology Score (SAPS) II, our score yielded a better prediction of ICU mortality in the receiver operator characteristics (ROC) analysis (AUC = 0.84 vs AUC = 0.73 in the validation cohort). Factors predicting decreased survival after ICU discharge were as follows: relapse or refractory disease, previous allogeneic stem cell transplantation, time between hospital admission and ICU admission, time spent in ICU, impaired diuresis, Glasgow Coma Scale < 8 and hematocrit of ≥25% at ICU admission. Based on these factors, an ICU survival score was created and used for risk stratification into three risk groups. This stratification discriminated distinct survival rates after ICU discharge. Conclusions: Based on data from a large multicenter cohort, we identified and validated relevant risk predictors, which provided a basis for two

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scores distinguishing between survival differences both in the ICU as well as after ICU discharge. However, while these scores might aid the prognostication of patients with AML treated in the ICU, decisions about initiating or pursuing intensive treatment must not rely solely on the results of these scores. This study should encourage further prospective analyses. Disclosure: No conflict of interest disclosed. P198

Comparison of ciprofloxacin and colistin as prophylactic antibiotics for the treatment of chemotherapy-induced neutropenia in patients with acute myeloid leukaemia: Two parallel patient cohorts treated in a single centre Pohlen M.1, Marx J.1, Melmann A.2, Becker K.3, Mesters R.M.1, Mikesch J.-H.1, Schliemann C.1, Lenz G.1, Müller-Tidow C.4, Büchner T.1, Krug U.5, Stelljes M.1, Karch H.2, Peters G.3, Gerth U.6, Görlich D.7, Berdel W.E.1 Universitätsklinikum Münster, Medizinische Klinik A – Hämatologie und Onkologie, Münster, Germany, 2Universitätsklinikum Münster, Institut für Hygiene, Münster, Germany, 3Universitätsklinikum Münster, Institut für Medizinische Mikrobiologie, Münster, Germany, 4Universitätsklinikum HalleWittenberg, Medizinische Klinik IV, Halle/Saale, Germany, 5Klinikum Leverkusen, Medizinische Klinik 3, Leverkusen, Germany, 6Universitätsklinikum Münster, Medizinische Klinik D – Innere Medizin, Nephrologie und Rheumatologie, Münster, Germany, 7Universitätsklinikum Münster, Institut für Biometrie und Statistik, Münster, Germany 1

Introduction: Patients with acute myeloid leukaemia (AML) are at high risk for bacterial infections during chemotherapy-related neutropenia. However, the use of antibiotic prophylaxis in afebrile neutropenic AML patients is controversial. Methods: This was a retrospective evaluation of 172 AML patients who were randomly allocated to receive antibiotic prophylaxis with colistin or ciprofloxacin during chemotherapy-related neutropenia (expected duration of >7 days). A total of 322 treatment courses with at least one chemotherapy course per stay were analyzed. During a total of 44 courses (14.9%) in 35 patients, antibiotic prophylaxis was not administered. The remaining 138 patients received antibiotic prophylaxis over 252 treatment courses; in detail, 57 patients received ciprofloxacin (in 130 courses), and 72 patients received colistin (in 122 courses). The differences between groups were analyzed through statistical methods capable of modelling repeated measurements, specifically generalized estimation equations (GEEs) were applied. Results: The infection rate was reduced from 88.6% to 74.2% through antibiotic prophylaxis (vs. without prophylaxis, p = 0.04). A comparison of both antibiotic drugs revealed a trend towards fewer infections associated with ciprofloxacin prophylaxis (69.2% vs. 79.5% in the colistin group, p = 0.07), as determined by univariate analysis. This result was confirmed through multivariate analysis (odds ratio [OR] 0.475, 95% confidence interval [CI] 0.236 - 0.958; p = 0.041). The prophylactic agents did not differ with regard to the microbiological findings (p = 0.6, n.s.). The risk factors for higher infection rates were the presence of a central venous catheter (p < 0.0001), mucositis grade III/IV (p = 0.0039), and induction/relapse courses (vs. consolidation, p < 0.0001). Conclusions: Ciprofloxacin prophylaxis appears to be of particular benefit during induction and relapse chemotherapy for AML, but it may be safely replaced by colistin during consolidation cycles of AML therapy. The selection of prophylactic agents should take into account variables like therapy stage (consolidation vs. induction/relapse), the risk of developing mucositis, and the local distribution of resistant pathogens. Disclosure: No conflict of interest disclosed.

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Pan-mammalian target of rapamycin (mTOR) inhibitor PP242 synergistically enhance ABT-737 induced apoptosis in acute myeloid leukemia (AML) cells Ahmed F.1, Ilyas A.1 King Abdulaziz University, Center of Excellence in Genomic Medicine Research, Djidda, Saudi Arabia 1

Pro-survival members of the BCL-2 family proteins are known to be overexpressed in hematological malignancies, including acute myeloid leukemia (AML). These pro-survival BCL-2 members that include BCL2, BCL-xL, MCL-1 and BCL-w gives the cancer cells a selective survival advantage and resistance against a broad range of apoptotic stimuli. Therefore, several agents targeting pro-survival BCL-2 proteins have been developed and are being currently tested in the clinical settings. ABT-737 and its orally available derivative ABT-263, and more recently ABT-199 have displayed promising early results in CLL and AML. However, it is unlikely that these inhibitors will be sufficient as single agents. Finding combinations that could potentiate the effects of BCL-2 inhibitors has become imperative. Here, we screened two PI3K/AKT/mTOR pathways inhibitors for possible synergistic killing of multiple AML cell lines and patient derived primary cells. AML cells were cultured and incubated with dual PI3K/mTOR inhibitor PKI402 or mTORC1&2 inhibitor PP242 and combined with BCL2 specific inhibitor ABT-737. Cell proliferation assays were performed to assess the cytotoxicity. Chou-Talalay method of multi-drug combination was used to determine combination index (CI) values as a measure for the interaction between two drugs. Flow cytometry was used to study apoptosis, mitochondrial potential and Caspase 3 activity. Intracellular mapping of target proteins was performed by flow cytometry. PKI-402, demonstrated additive effect in combination with ABT-737 in HL60, TF1 and K562 cells and showed synergistic effect in MV-411 and NB4 cells. PP242, the dual mTORC1/2 inhibitor, demonstrated synergistic inhibition of AML cell proliferation with ABT-737 and induced apoptosis in multiple AML cell lines. The increased induction of apoptosis by PP242 in combination with ABT-737 was associated with increased caspase 3 activity and depolarization of mitochondrial membrane. Phospho protein analysis by flow cytometry revealed that combination of PP242 and ABT737 significantly down regulated phosphorylation of BCL-2 and ribosomal protein S6 to induce apoptosis. Ex vivo treatment of AML patient derived blast cells revealed enhancement of ABT-737 induced apoptosis by PP242. In conclusion, the combination of pan-mTOR inhibitor PP242 with ABT737 represent a potentially promising approach in the treatment of AML and has the potential to overcome the limited efficacy of single agents. Disclosure: No conflict of interest disclosed. P200

Biomodulatory therapy with low-dose azacitidine, pioglitazone and ATRA induces myeloid differentiation and phagocytosis in primary AML blasts Klobuch S.1, Steinberg T.1, Reichle A.1, Herr W.1,2, Thomas S.1,2 Universitätsklinikum Regensburg, Klinik und Poliklinik für Innere Medizin III, Regensburg, Germany, 2Regensburger Centrum für Interventionelle Immunologie, Regensburg, Germany 1

Clinical outcome of elderly patients with chemorefractory acute myeloid leukemia (AML) is very poor and allogeneic hematopoietic stem cell transplantation is rarely feasible. We recently described in a small cohort of 5 chemorefractory elderly AML patients a novel biomodulatory therapy consisting of low-dose 5-azacitidine (AZA) in combination with the PPARγ ligand pioglitazone (PGZ) and all-trans retinoic acid (ATRA). The therapy induced complete remissions in 4/5 patients and was associated with strong myeloid differentiation and a rapid increase of peripheral blood neutrophils. Interestingly, peripheral blood neutrophils expressed the same AML-specific mutations than primary AML blasts, suggesting

Abstracts

that the early increase of neutrophils observed during treatment included a considerable fraction of differentiated blasts. To analyze these clinical observations in AML cultures in vitro, we treated primary AML blasts from different patients with AZA, PGZ and ATRA and subsequently evaluated myeloid cell differentiation by means of morphological changes, the expression of differentiation antigens and phagocytosis capacity. In samples from 4 of 8 patients we detected a 2 to 4-fold increase of cells showing morphological features of myeloid differentiation (i.e. nuclear lobulation), an increase of CD11b surface expression as well as growth control during 14 days of treatment with AZA/ PGZ/ATRA. However, ATRA monotherapy was less efficient to induce myeloid differentiation in AML blasts from the same patients, suggesting that combination therapy might act synergistically on leukemic differentiation. Interestingly, differentiated cells that arise during in vitro treatment with AZA/PGZ/ATRA also showed an increase in phagocytosis, as demonstrated by a 2-fold increase in the uptake of GFP transfected E. coli as well as ROS production. Again, ATRA treatment was less efficient to induce phagocytosis in analyzed AML samples. We conclude that biomodulatory combination therapy with low-dose AZA, pioglitazone and ATRA can be sufficient to drive differentiation of primary AML blasts into functional neutrophils, which may lower the neutropenia associated infection rates in AML patients. Clinical impact on infections rates as well as overall survival of low-dose AZA/PGZ/ ATRA treatment will be further investigated in a planned randomized clinical trial on chemorefractory elderly AML patients. Disclosure: No conflict of interest disclosed.

Posterdiskussion

Chronische myeloische Leukämie P201

Major route additional chromosomal aberrations (ACA) precede increase of blasts in chronic myeloid leukemia (CML) independent of therapy. An analysis of CML studies III, IIIA and IV Voskanyan A.1, Dietz C.T.2, Fabarius A.C.1, Lauseker M.3, Saußele S.1, Kalmanti L.4, Rinaldetti S.1, Haferlach C.5, Pfirrmann M.3, Hasford J.3, Baerlocher G.M.6, Hochhaus A.7, Baccarani M.8, Hehlmann R.1, for the SAKK and the German CML Study Group Medizinische Fakultät Mannheim, Universität Heidelberg, III. Medizinische Klinik, Mannheim, Germany, 2IHO Institute for Hematology and Oncology, Mannheim, Germany, 3Ludwig-Maximilians Universität München, Institut für medizinische Informationsverarbeitung, Biometrie und Epidemiologie, München, Germany, 4DKD HELIOS Klinik Wiesbaden, Zentrum für Blutstammzell- und Knochenmarktransplantation, Wiesbaden, Germany, 5 MLL Münchner Leukämielabor, München, Germany, 6Inselspital Bern, Universitätsklinik für Hämatologie und Hämatologisches Zentrallabor, Bern, Switzerland, 7Universitätsklinikum Jena, Abteilung Hämatologie/Onkologie, Klinik für Innere Medizin II, Jena, Germany, 8University of Bologna, Department of Hematology and Oncology, Bologna, Italy 1

Progression of CML to blast crisis is poorly understood. The aim was to determine the possible impact of major route and non-major route ACA in Ph+ cells in the course of disease on progression of CML. Patients (pts.) with primary treatments interferon-alpha (CML studies III and IIIA) and imatinib (CML study IV) were analyzed for correlation of ACA with blast increase (BI). 1287 pts. recruited to CML-studies III/IIIA were evaluable with a 10-year survival of 48% and 61%, respectively, of whom 258 (20%) progressed to blast crisis (BC). 195 pts. displayed ACA, 45 at baseline. 109 pts. showed major route ACA. 1536 pts. recruited to CML-study IV were evaluable with a 10-year survival of 83% of whom 81 (5,3%) progressed to BC. 183 pts. displayed ACA,105 at baseline. 86 pts. showed major route ACA. In a univariate analysis of CML studies III/IIIA, pts. with ACA had a hazard ratio (HR) for BI ranging between 2.45–3.27 (p < 0.001) for blast

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counts from 1% to > 30%compared with pts. without ACA. For major route ACA, HRs of 3.5–4.5 were found. For non-major route ACA HRs were 1.7-2.2 (p < 0.001). In the same model with CML study IV, pts. with ACA had a HR for BI ranging between 4.5-8.9 (p < 0.001) compared with pts. without ACA. For major route ACA, HRs of 13.1-27.8 (p < 0.001) were found. For non-major route ACA HRs were 1.05-1.07 (p = n.s.). In both studies the cumulative incidences of BI by 5 years after diagnosis of major route ACA was 40% (Fig.1). In CML III/IIIA pts. cumulative incidences of BI by 5 years after diagnosis of non-major route ACA was 25-30%, in CML IV pts. 1- 5% (Fig.2).

Fig. 1.

Fig. 2.

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Protein-tyrosine phosphatases modulate therapy response in CML cells Drube J.1, Albert B.V.1, Pfirrmann M.2, Ernst T.3, Hochhaus A.3, Böhmer F.-D.1 Universitätsklinikum Jena, Institut für Molekulare Zellbiologie, CMB, Jena, Germany, 2Ludwig-Maximilians-Universität München, Institut für Medizinische Informationsverarbeitung, Biometrie und Epidemiologie (IBE), München, Germany, 3Universitätsklinikum Jena, Klinik für Innere Medizin II, Abteilung Hämatologie und Internistische Onkologie, Jena, Germany 1

Introduction: Most patients with chronic myelogenous leukemia (CML) can be successfully treated with tyrosine kinase inhibitors (TKIs) targeting the protein tyrosine kinase (PTK) BCR-ABL, but some do not reach molecular remission on the 4log level (MR4) or not even a 3log reduction (MMR). The counteractors of PTK are protein tyrosine phosphatases (PTPs). It is likely, that expression levels of specific PTPs modify the response to BCR-ABL inhibitors and that they have an impact on treatment outcome. Methods: A qPCR-platform for the expression analysis of 38 PTPs was established, and the PTP expression status in leukocytes of 66 newly diagnosed, untreated CML patients, was analyzed. This data for each individual PTP was correlated with BCR-ABLIS after 9 months of nilotinib treatment (2*300 mg/d) using logistic regression analysis. Several PTPs, including CD45, PTPRG, and SHP-1 were selected for further functional analysis using cell lines with shRNA-mediated depletion, CRISPR/Cas9 knockout, and stable re-/overexpression. The engineered lines were treated with TKI and the IC50 values were determined. Additionally we investigated the effect of BCR-ABL on PTP expression in a set of newly created control-, BCR-ABL-, and TKI resistant BCR-ABLT315I-expressing cells. Results: Comparing patients with MR4 or better (n = 36) to patients not having a log4 reduction (n = 30) after 9 months of therapy, a higher expression of CD45 significantly increased the probability of MR4 at 9 months (p = 0.007). A significant association was also found for higher PTPRG levels (p = 0.012). Interestingly, cell culture studies revealed lowering of the nilotinib IC50 in case of the CD45 depletion, indicating increased responsiveness. In contrast, SHP-1 deficient cells were more resistant to nilotinib compared to control cells. PTPRG (encoding PTPγ) was identified as a novel BCR-ABL regulated PTP gene. We observed an increase of PTPRG expression in the BCR-ABL and BCR-ABL-T315I-expressing cells, that decreased to control-levels upon nilotinib-treatment in case of the BCR-ABL cells, but not in the T315I-mutant cell lines. The role of PTPRG levels and the mechanisms of PTP-mediated modulation of TKI responses are currently being investigated. Conclusions: Several PTPs correlate in expression with the therapy response to nilotinib and appear to causally affect responses in cell line models. Understanding the underlying mechanisms may aid in further optimizing TKI treatment. Disclosure: No conflict of interest disclosed.

ACA, particularly major route ACA preceded an increase of blasts. For pts. with major route ACA the 5-year-probability of a BI was 40% independent of treatment type. The impact of non-major route ACA on BI appeared to be eliminated by imatinib in contrast to IFN treatment. Disclosure: No conflict of interest disclosed.

P203

BCR-ABL independent gene mutations in children and adolescents with chronic myeloid leukemia Busch M.1, Rinke J.1, Schäfer V.1, Waldau A.1, Ernst J.2, Hochhaus A.1, Gruhn B.2, Ernst T.1 Abteilung Hämatologie und Internistische Onkologie, Klinik für Innere Medizin II, Universitätsklinikum Jena, Jena, Germany, 2Klinik für Kinder- und Jugendmedizin, Universitätsklinikum Jena, Jena, Germany 1

Introduction: Chronic myeloid leukemia (CML) in children and adolescents is rare. Many diagnostic standards and therapy options have been adapted from adult CML. We have previously identified BCR-ABL independent gene mutations in 33% of newly diagnosed adult CML patients that may function as important cofactors in the evolution of CML (Schmidt M, Rinke J, et al. Leukemia 2014; 28(12): 2292-2299). Here we

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sought to investigate whether BCR-ABL independent gene mutations can also be observed in children and adolescents with CML. Methods: The cohort included 20 children and adolescents (male, n = 14; median age: 14 years, range: 10 months to 27 years) suffering from CML. Patients’ DNA was isolated from peripheral blood leukocytes obtained at diagnosis. Targeted deep next-generation sequencing (NGS) was applied to analyze a panel of 30 commonly mutated genes in myeloid disorders, affecting signal transduction, transcription factors, epigenetic regulation, RNA splicing and the cohesin complex. Results: BCR-ABL independent gene mutations were detected in 8 of 20 patients (40%). In total, 14 mutations were detected in 7 different genes, namely ASXL1, DNMT3A, EZH2, JAK2, KRAS, STAG2, and UTX. Ten mutations affected epigenetic modifier genes. Mutations included 10 missense mutations, 2 nonsense mutations and 2 deletions. To investigate whether mutations originated in the myeloid lineage or are likely to be germline, purified T-cell DNA of selected patients will be analyzed until the meeting. Conclusions: Analogous to adult CML, BCR-ABL independent gene mutations can also be frequently found in children and adolescents with CML. This implicates that BCR-ABL independent gene mutations are not age-related events. Thus, such molecular aberrations may play an important role in the evolution and persistence of the disease and may affect therapy in both children and adults with CML. Our findings underline a potential multistep pathogenesis and the important role of epigenetic events in CML biology. Disclosure: No conflict of interest disclosed. P204

The Jak2-inhibitor Pacritinib with wide kinome-profile overcomes cytokine-mediated and mutation-driven resistance in CML cell lines Hammersen J.1, Haase J.1, Becker C.1, Clement J.1, Hochhaus A.1, La Rosée P.1 Universitätsklinikum Jena, Klinik für Innere Medizin II, Hämatologie/Onkologie, Jena, Germany 1

Introduction: Targeting the JAK/STAT-pathway in chronic myelogenous leukaemia (CML) interfers with the ability of BCR-ABL independent survival attributed to pathologic stem- and progenitor cells (SPC). However, this primary resistance of CML-SPC is not driven “one way” via JAK-signaling, but regulated by multiple kinase-signals. Pacritinib (PA) is a new JAK2/FLT3 inhibitor with a wide kinome profile. Aim of this study was to investigate the effects of PA in CML cell lines and primary CML SPCs in combination with Nilotinib (NI). Material/Methods: Cells: BCR-ABL transfected murine Ba/F3p210 wildtype/TKI-resistant mutants & human M07p210. Incubation +/- cytokines IL3/GM-CSF. Antiproliferative effects tested in MTS assays, signaling by Western blot (WB), synergism analysis by the method of Chou (Combination-Index (CI) >1.1: antagonism; < 0.9: synergism). Methocult-assays (MA) to assess effects on the clonogenic capacity of primary CD34 sorted cells from CML patients. Results: IL3-exposed Ba/F3p210 lost sensitivity to NI, which served as model for primary cytokine-driven resistance. It was overcome by adding PA: IC50 IL3+NI (mono) > 64nM vs. IC50 IL3+NI+PA[500nM] 4nM. M07p210 reacted analogously. NI&PA showed synergism in both cell lines (CI = 0.7/0.5). Ba/F3 transfected with BCR-ABL mutants G250E or T315I showed resistance to NI [IC50>64nM], a model for mutation induced secondary resistance. PA alone was active in unmutated and resistance mutation expressing BaF3 cells (IC50 non-mutant p210: 500nM; G250E: 638nM; T315I: 785nM). In Ba/F3 PA was more active in p210 transfected than in parental cells IC50 = 800nM, suggesting partially BCR-ABL-dependent activity. Combination treatment in IL3-depleted media showed an antagonistic effect (CI = 1.7). Cytokine-mediated activation of STAT5 is inhibited by PA [500nM]/PA+NI [500nM+20nM], as detected by WB. Incubation of NI/PA/NI+PA (2000/750/2000+750nM) in MA from 3 pa-

Abstracts

tients lead to reduction of CFU-GM colonies of 47 / 72 / 93% with high inter-patient variability. Discussion: PA shows antiproliferative effects in CML cell lines & primary CML SPCs. It is able to overcome cytokine driven TKI resistance, partially explained by inhibition of STAT5 phosphorylation. Synergism of PA/NI and its activity against NI-resistant BCR-ABL mutants can be explained by interference with cytokine signaling and a wide kinome profile. PA is identified as potential partner in combination therapy of CML to prevent and overcome resistance. Disclosure: No conflict of interest disclosed. P205

Resistance in chronic myeloid leukemia: Therapeutic targeting of escape via CSF2RB Becker C.1, Poser I.1, Wohlmann A.2, Clement J.1, Friedrich K.-H.2, Hochhaus A.1, La Rosée P.1 Universitätsklinikum Jena, Hämatologie und Internistische Onkologie, Jena, Germany, 2Universitätsklinikum Jena, Institut für Biochemie II, Jena, Germany 1

Introduction: Persistence of minimal residual disease constitutes a risk of disease recurrence in patients with chronic myelogenous leukaemia (CML) treated with Abl-inhibitors. Persistent BCR-ABL+ leukemic stem and progenitor cells are promoted by bone marrow stroma cytokines such as interleukin 3 (IL-3) and granulocyte/macrophage-colony stimulating factor (GM-CSF) mediating BCR-ABL-independent survival via CSF2RB. We demonstrate upregulation of CSF2RB in BCR-ABL-transformed cells and primary cell samples. Disruption of the CSF2RB axis by the Janus kinase1/2-inhibitor ruxolitinib overcomes cytokine-mediated resistance in vitro as seen in an indepth molecular and functional analysis. Methods: Gene expression analysis of CSF2RB was performed in M07p210 cells and patient derived BCR-ABL+ CML cells from bone marrow in response to nilotinib. Functional relevance of CSF2RB-mediated rescue of leukemic cells was investigated by gene knockdown of CSF2RB using siRNA in human M07p210. Viability and colony formation of BCRABL positive CML CD34+ progenitor cells in cytokine enriched nilotinib or dasatinib with or without ruxolitinib supplemented media were analyzed in vitro. Results: Upon TKI treatment of M07p210 cells and CML CD34+ progenitor cells, compensatory upregulation of CSF2RB exert distinct survival signals including cytokine-induced STAT5 activation, despite effective BCR-ABL-inhibition. CSF2RB is regulated on the transcriptional level as demonstrated by quantitative RT-PCR. Knockdown of CSF2RB or combination treatment of M07p210 and treatment naïve CML CD34+ progenitor cells with nilotinib or dasatinib and ruxolitinib overcomes cytokine-mediated TKI-resistance as demonstrated by additive growth inhibition in the presence of GM-CSF. Increased mRNA-expression of CSF2RB is demonstrated in bone marrow harvested at 3 months of nilotinib (n = 8). No correlation with molecular response could be demonstrated in this exploratory cohort. Conclusion: Upregulation of CSF2RB in response to TKIs is a consistent finding in CML in vitro and ex vivo with high pre-clinical activity of combined ruxolitinib. Exploratory analysis of CSF2RB as biomarker for response fails to demonstrate a robust relationship in individual patients. Further analysis is ongoing. Disclosure: No conflict of interest disclosed. P206

BCR-ABL suppresses canonical BMP-signalling Knobloch U.1, Elsner M.1, La Rosée P.1, Hochhaus A.1, Clement J.H.1 Klinik für Innere Medizin II, Universitätsklinikum Jena, Hämatologie und Internistische Onkologie, Jena, Germany 1

Introduction: Chronic myeloid leukemia (CML) is a myeloproliferative disease of pluripotent hematopoietic stem cells and is characterized by the Philadelphia chromosome and the fusion protein BCR-ABL. Bone mor-

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phogenetic proteins (BMP) are essential regulators of differentiation and cell death during embryogenesis as well as in adults. The aim of this study was to evaluate the presence of key components of the BMP signalling cascade in BCR-ABL-positive cells and their responsiveness for BMP signals. Methods: The cell lines M07e (acute megakaryoblast leukemia), M07ep210 (stably overexpressing bcr-abl) and K562 (CML, blast crisis) were cultivated with RPMI1640 + 20% FCS under standard conditions; for M07e supplemented with GM-CSF. Incubation experiments were performed with BMP-2, BMP-4, BMP-6 or the BMP inhibitor dorsomorphin. Gene expression was measured by reverse transcriptase PCR and qPCR. Protein expression and activity was estimated by immunoblotting. Cell vitality was monitored by AnnexinV/Propidium iodide staining. Results: PCR analysis showed that most of the BMPs, their receptors and molecules involved in canonical BMP signalling are expressed in the investigated cell lines on a comparable level except BMP-2 (high expression in M07e) and Activin receptor 2A (ACVR2A) (low expression in M07e). The most dramatic changes were observed for Smad1. Its expression was reduced 5-fold in K562 compared to M07e and was undetectable in M07ep210. Thus, a reduced sensitivity for BMPs acting via the canonical Smad signalling pathway was expected for bcr-abl-expressing cells. Interestingly, the well-known BMP target gene ID1 was highly expressed in bcr-abl-expressing cells, but not in M07e. On the protein level, ID1 was not regulated by BMP-2, BMP-4 or BMP-6. These observations point to alternate activation of ID1 expression and to a more undifferentiated state of the bcr-abl-expressing cells. BMPs are involved in regulation of apoptosis. Nether the application of BMP-2 nor the BMP inhibitor dorsomorphin affected the viability of the bcr-abl-expressing cells. In contrast, BMP-2 enhanced the vitality of M07e slightly, but dorsomorphin reduced vitality more than 6-fold compared to untreated cells. Conclusion: The canonical BMP signalling via the Smad pathway is suppressed in the presence of BCR-ABL. This may keep the cells in a more undifferentiated state and protect them from pro-apoptotic signals transmitted via receptor-serine/threonine kinases. Disclosure: No conflict of interest disclosed. P207

European survey on the assessment of deep molecular response in chronic phase CML patients after at least two years of therapy with tyrosine kinase inhibitors (TKI) (EUREKA). Schenk T.1, Lange T.2, Saussele S.3, Pott C.4, Ernst T.1, Cross N.C.P.5, Hochhaus A.1 Klinik für Innere Medizin II, Universitätsklinikum, Jena, Germany, 2Klinik für Hämatologie und internistische Onkologie, Asklepios-Klinikum, Weissenfels, Germany, 3III. Medizinische Klinik, Universitätsmedizin, Mannheim, Germany, 4 2. Med. Klinik, Universitätsklinikum Schleswig-Holstein, Campus Kiel, Kiel, Germany, 5Wessex Regional Genetics Laboratory, Salisbury District Hospital, Salisbury, United Kingdom 1

Introduction: The advent of highly effective therapies has changed the natural history of chronic myeloid leukemia (CML). The European LeukemiaNet (ELN) recommends quantification of BCR-ABL transcripts by real-time quantitative polymerase chain reaction (RQ-PCR) every 3 mo during TKI therapy. Recently, interest has focused on the assessment of deep molecular response (DMR, MR4, MR4.5, MR5) because a proportion of patients (pts) maintain remission after treatment stop. Molecular (mol) data monitored with a sensitive and standardized assay collected systematically do not exist so far outside of a clinical trial setting. Methods: The purpose of this ongoing registry is to collect data on the standardized assessment of molecular response in the context of real life practice. BCR-ABL transcript levels after >2 yrs of TKI therapy are evaluated for DMR and its impact on pts’ management. Since standardized mol monitoring is a prerequisite for treatment discontinuation, central surveillance of the performance of the participating labs has been introduced.

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Results: Between 2014-16, 1768 samples were sent after pts’ informed consent to 9 laboratories in 6 countries (D, I, H, CZ, HR, BIH). Pts’ median age was 59 yrs (range, 19-90), 55.5% of pts were male. Transcript type was b2a2 in 460 (26.0%), b3a2 in 685 (38.7%), b2a2/b3a2 in 47 cases (2.7%). 10 samples showed atypical transcripts (0.6%; e19a2, n = 5; e1a2, n = 3; b2a3, n = 2). In 555 cases (31.4%), no initial transcript type was reported. 1114 (63.0%), 451 (25.5%), and 187 (10.6%) samples were from pts on 1st, 2nd, or 3rd line therapies, respectively. Line of therapy was unknown in 16 cases. Current therapy was imatinib, n = 939 (53.1%), nilotinib, n = 517 (29.2%), dasatinib, n = 180 (10.2%), ponatinib, n = 9 (0.5%), bosutinib, n = 7 (0.4%), other, n = 12 (0.7%), unknown, n = 104 (5.9%). Response levels were: No MMR, n = 102 (5.8%); MMR, n = 283 (16.0%); MR4, n = 411 (23.2%); MR4.5, n = 538 (30.4%); MR5, n = 392 (22.2%). RQPCR failed in 42 cases (2.4%). Previous judgement as “complete” mol response was specified in 47 cases. Conclusions: Improving the monitoring of deeper and sustained mol responses is critical for the optimal management of BCR-ABL+ CML patients and will assist to define parameters for treatment discontinuation. Multicenter DMR assessment is feasible in the context of real life clinical practice. Information on the BCR-ABL transcript type at diagnosis is crucial to accurately follow up pts’ response on or after therapy. Disclosure: Thomas Schenk: No conflict of interest disclosed. Andreas Hochhaus: Financing of Scientific Research: Novartis, BMS, Pfizer, Ariad; Expert Testimony: Novartis, BMS, Pfizer, Ariad P208

High molecular response rates in patients with chronic myeloid leukemia – a retrospective health care analysis of more than 1000 patients Tesch H.1, Jost P.2, Kisro J.3, Springer G.4 Centrum für Hämatologie und Onkologie Bethanien, Frankfurt, Germany, 2III. Medizinische Klinik, Klinikum rechts der Isar, München, Germany, 3Lübecker Onkologische Schwerpunktpraxis, Lübeck, Germany, 4Praxis für Onkologie, Hämatologie und Palliativmedizin Stuttgart, Stuttgart, Germany 1

Introduction: Chronic myeloid leukemia (CML) is a clonal myeloproliferative disorder of the hematopoietic stem cells. The aim of this retrospective analysis was to analyze guideline adherence and treatment results in German CML centers under real life condition. Methods: The quality of molecular monitoring and health care for CML patients was evaluated by a questionnaire according to current CML recommendations. With data of more than 1000 patients from 49 centers (7 clinics, 42 oncology practices) distributed all over Germany, it is representative for the health care situation of CML patients in German oncology centers. Results: For the initial analysis, data from 1002 patients registered in the period from 08-2013 to 09-2015 were analyzed. Patients characteristics at diagnosis such as age, sex and phase of disease at diagnosis were similar to literature. 24.8% of patients were enrolled in first-line therapy CML studies. While cytogenetic analysis was not performed in 68.1% of patients during the last 12 months, quantitative PCR analysis was regularly performed in 92.5% of patients. Monitoring with quantitative PCR analysis was reported for 59.8% of patients every 3 months until reaching the BCR-ABL value of < 0.1. The majority of patients (60.8%) received first-line therapy with tyrosine kinase inhibitors (TKIs), whereas 26.2% were treated in second-line therapy. Response parameters corresponded to published data of clinical trials. In 82.9% of the patients, hematological response was maintained during therapy. Cytogenetic response was maintained in 63.2% and molecular response in 70.7% of patients. A high proportion of 69.5% of patients had achieved major molecular response (MR3.0) and 58.4% deep molecular response (MR4.0) at the time point of last visit. In an extension of the analysis, further data of 300 patients will be included from the ongoing project. Conclusion: As the cohort investigated covers about 1/10 of the CML patients in Germany the data from this extensive retrospective chart analysis complements current knowledge about management of CML patients in

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Germany outside of clinical trials in a real-world setting. Importantly current CML recommendations are being followed in general and therefore demonstrate a high standard in molecular monitoring of CML patients in Germany. Disclosure: Hans Tesch: Employment or Leadership Position: Niedergelassener Onkologe im Centrum für Hämatologie und Onkologie Bethanien Frankfurt.; Advisory Role: Novartis, BMS; Financing of Scientific Research: Novartis, BMS Gregor Springer: Financing of Scientific Research: Novartis; Other Financial Relationships: Novartis, BMS P209

Molecular monitoring in assessing minimal residual disease and risk of progression in chronic phase CML: our data at King Fahad Medical City Alsohaibani L.1, Altahan R.1, Alshehri N.1, Tailor I.K.1, Motabi I.1, Almishari M.1, Tashkandi S.1, Peer Zada A.A.1 King Fahad Medical City, Riyadh, Saudi Arabia

Background: The World Health Organization and European Leukemia Net classification has set up criteria for the definition of the chronic phase (CP), accelerated phase (AP), and blast crisis (BC) of CML. In CP-CML patients on therapy, the reduced disease burden is an important prognostic indicator and minimal response to therapy or relapse suggests a therapy change. Monitoring minimal residual disease in CP-CML in view of its clinical and therapeutic outcome therefore, becomes essential. Aim: We sought to assess levels of MRD in CP-CML patients by measuring the levels of BCR-ABL1 p210 fusion gene using an automated real-time RT-PCR test, the Xpert BCR-ABL (p210) Monitor Assay and assess the progression free survival, and the rate of relapse in our patients. We also assessed the rate of TKI response in CP-CML cases. Cytogenetic analysis (FISH) on bone marrow or peripheral blood samples was used to assess cytogenetic remission (or not) in all CP-CML cases. Results: We describe here 51 CP-CML patients consisting of 29 females (~57%) with a median age of 44 ± 4.9 years (95% CI 26.6- 62.39) and 22 males (43%) with a median age of 36 ± 7.7 years. In patients with cytogenetic remission, BCR-ABL1 p210 level was detectable in 30 (30/51, 58.8%) and undetectable (< LOD) in 16 (16/51, 31.3%). 5 patients showed very high levels of p210 either due to relapse, intolerance, or compliance issues. Of the 30 CP-CML patients, 21 (70%) patients revealed BCR-ABL1/ABL ratio < 0.1% IS (MMR) are in continuous remission while 9 (30%) patients showed >0.1% IS p210 levels and did not achieve MMR. In response to TKI treatment, 48 (94%) CP-CML patients received imatinib as first-line, of whom 35 (72.9%) achieved molecular remission while 13 (27%) revealed treatment failure based on molecular data. Desatinib and nilotinib as second-line were effective in reducing p210 levels (MMR) in patients who failed imatinib treatment. 2 patients received desatinib as first-line and both (100%) achieved molecular remission. 25 (/48, 52%) CP-CML patients showed >5 years of progression free survival with imatinib with respect to p210 levels (continuous MMR). Conclusion: In summary, depending upon the individual patient co-morbidities, all TKIs (Imatinib, desatinib, Nilotinib) showed good molecular response in CML patients albeit with some degree of intolerance. In CPCML patients who failed imatinib treatment, desatinib and nilotinib were effective in achieving molecular milestones. Disclosure: No conflict of interest disclosed.

Abstracts

P210

Results of the non-interventional TARGET study: Efficacy and safety of nilotinib in CML patients (pts) failing prior therapy in routine healthcare Dengler J.1, Müller M.C.2, Buß E.3, le Coutre P.4, Stegelmann F.5, Ulshöfer T.6, Sauer A.7, Schardt C.8, Reichert D.9, Schwinger U.10, Grunewald R.11, Waller C.12, Meincke M.13, Rupprecht S.13, Tesch H.11 Onkologische Schwerpunktpraxis, Heilbronn, Germany, 2Universitätsklinikum Mannheim, Mannheim, Germany, 3Universitätsklinikum Heidelberg, Heidelberg, Germany, 4Charité – Universitätsmedizin Berlin, Berlin, Germany, 5 Universitätsklinikum Ulm, Ulm, Germany, 6Schwerpunktpraxis für Hämatologie und Onkologie, Ludwigsburg, Germany, 7Medizinisches Versorgungszentrum für Blut- und Krebserkrankungen, Potsdam, Germany, 8Onkologische Praxis und Tagesklinik, Gelsenkirchen, Germany, 9Gemeinschaftspraxis für Hämatologie und Onkologie, Westerstede, Germany, 10Onkologikum, Stuttgart, Germany, 11 Onkologie Bethanien, Frankfurt a.M., Germany, 12Universitätsklinikum Freiburg, Freiburg, Germany, 13Novartis Pharma GmbH, Nürnberg, Germany 1

Introduction: Nilotinib (NI) as a potent and highly selective BCR-ABL tyrosine kinase inhibitor is indicated for Ph+ CML pts in CP and AP with intolerance to or resistance of prior therapy including imatinib (IM) as well as for de novo Ph+ CML in CP. Methods: Follow-up analysis of an observational study of NI in 479 pts with Ph+ CML resistant to or intolerant of prior treatment within routine clinical management in 156 centres in Germany (Jan 2008-Mar 2016). Results: The median age of pts was 65.1 yrs, with 7.9% being older than 80 yrs. 99.2% (0.6%) presented in CP (AP)(phase missing in 1pt). 91% had a good performance status (Karnofsky index ≤ 1). 92.7% of all pts were pretreated with IM (any dose). Further prior drug treatments were chemotherapy (23.4%), IFN (16.3%), dasatinib (19%) and other unspecified drugs (12.5%). Three pts had received SCT in the past. 56.6%/45% (n = 129) were treated with NI mostly due to resistance/intolerance against IM and 26.4%/75.8% (n = 91) against dasatinib, respectively (multiple responses possible). At initial visit, a dose of 800 mg NI/d was prescribed in 50.7% and 600 mg/d in 27.1%. Median duration of NI was 633 days at the data cut-off for this analysis. Remission status at study entry was 60.8% in CHR, 42.2% in MCyR/26.3% in CCyR (missing data in 20.5%), 30.5%/10.6% in MMR/CMR. These responses improved significantly under NI, reaching cumulative incidences of CHR, MMR and CMR of 90.2%, 63% and 35.3%, respectively. Of note, cytogenetic response improved as well but is not conclusive enough due to missing examinations (87.7% after 12 months). Dose reduction or therapy interruption at any time occurred in 29.6% or 17%. 77.7% experienced at least one AE during the observation period which was considered serious in 15.2%. 42.1% of pts with documented final visit (n = 409) prematurely discontinued the study. Hematologic toxicity / non-hematologic toxicity were observed in 12.7%/41.1% of pts. The most frequently reported AEs (AEs ≥5%) were skin reactions (pruritus 11.7%, rash 9.6%, alopecia 6.7%), fatigue (10.9%), thrombocytopenia (6.9%), anaemia (5.6%), arthralgia (5.4%), dyspnoea (5.2%), nausea (6.9%), and upper abdominal pain (6.1%) as well as headache (9.8%). Coronary disease occurred in 5.7% of pts, whereas peripheral arterial occlusive disease was reported in 4.2%. Conclusions: This interim analysis supports NI as an efficacious and safe treatment option for CML pts with poor response or intolerance to a previous treatment. Disclosure: Jolanta Dengler: Employment or Leadership Position: Selbständige Ärztin; Financing of Scientific Research: Vortragshonorare der Firma Novartis Hans Tesch: Employment or Leadership Position: Niedergelassener Onkologe im Centrum für Hämatologie und Onkologie Bethanien Frankfurt; Advisory Role: Beratungstätigkeit Novartis; Financing of Scientific Research: Novartis

Oncol Res Treat 2016;39(suppl 3):1–104

CONTENTS AUTHOR INDEX

P211

P212

Efficacy and safety of Nilotinib in newly diagnosed Ph+ CML patients in chronic phase: Results of the 4th interim analysis of the non-interventional MOMENT II-study

DasPAQT: Treating patients with Chronic Myeloid Leukemia (CML) in Chronic Phase (CP) with Dasatinib: PCR-Monitoring, adherence, quality of life, therapy satisfaction (OMC 2014-I; BMS CA180-565) - an analysis of the first 150 patients

Lathan B.1, Sauer A.2, Tebbe S.3, Ulshöfer T.4, Lange E.5, Schulze M.6, Nusch A.7, Janssen J.8, Losem C.9, Meincke M.10, Rupprecht S.10, Tesch H.11 Gemeinschaftspraxis für Hämatologie und Onkologie, Dortmund, Germany, Medizinisches Versorgungszentrum für Blut- und Krebserkrankungen, Potsdam, Germany, 3Praxis für Hämatologie und Onkologie, Kassel, Germany, 4 Schwerpunktpraxis für Hämatologie und Onkologie, Ludwigsburg, Germany, 5 Evangelisches Krankenhaus, Hamm, Germany, 6Praxis und Tagesklinik für Hämatologie/Onkologie und Palliativmedizin, Zittau, Germany, 7Onkologische Praxis, Velbert, Germany, 8Gemeinschaftspraxis für Hämatologie und Onkologie, Westerstede, Germany, 9Praxis für Hämatologie und Onkologie, Neuss, Germany, 10Novartis Pharma GmbH, Nürnberg, Germany, 11Onkologie Bethanien, Frankfurt a.M., Germany 1 2

Introduction: Nilotinib (NI) is a potent and highly selective BCR-ABL TKI approved for treatment of newly diagnosed Ph+ CML pts in CP based on ENESTnd data showing improved treatment with higher molecular response rates vs. imatinib (IM). NI is also indicated for CP and AP Ph+ CML pts who failed prior therapy including IM. Methods: 4th interim analysis of a non-interventional study of NI in 362 pts with de novo Ph+ CML in CP within routine clinical management (Aug 2011- Mar 2016; 118 centres in Germany). Results: The median age was 58 yrs (17-88). 43.9% and 2.5% of the pts were older than 60 and 80 yrs. The median time since diagnosis of CML was 12 days (0-84). 93.1% of the pts had a good performance status (ECOG: ≤1; missing data in 3.9%). The median observation period was 586 days (4-1064).The median daily dose of NI was 600 mg (150-800 mg) with an initial NI daily dose of 600 mg in ~94% of the cases and a final NI daily dose of 600 mg >87% of the cases. There were 19.1% of pts with at least one therapy interruption and 15.5% of pts with at least one dose reduction. The most common reason for dosage adaption and/or therapy interruption were the occurrence of AEs (40,4%).At last visit 81.6% (of 315 pts with a hematologic examination) had a CHR (7% with missing data), 84.4% (of 32 pts with a cytogenetic examination) had a CCyR (96.9% with PCyR), 63.4% (of 268 pts with a molecular examination) had an MMR (1.5% with missing data). In the subgroup of pts with molecular response (MMR or better, n = 241) the median time to response was 191 days (56783). A premature treatment discontinuation took place in 18.2% of pts mostly due to AEs/non-hematologic toxicity, in 5 cases (7.6%) because of disease progression, which is mainly characterised by new BCR-ABL mutations. Altogether, 76.8% of pts experienced AEs. Hematologic toxicity was observed in 6,6% of pts (5.3% with thrombocytopenia), non-hematologic toxicities occurred in 37.3% of pts. The most frequently reported AEs were skin reactions (rash 8.8%, pruritus 8.3%, alopecia 5.8%), fatigue (9.1%), headache (5.5%), gastrointestinal symptoms (nausea 7.2%, upper abdominal pain 6.1%). No cardiac / vascular disorders were ≥ 2%. The most frequent biochemical abnormalities were increases in GGT (5.8%) and blood bilirubin (5.3%). Conclusions: NI is supported as an effective and safe treatment for newly diagnosed Ph+ CML pts in CP by these data from routine clinical management. Disclosure: Bernd Lathan: No conflict of interest disclosed. Hans Tesch: Employment or Leadership Position: Niedergelassener Onkologe im Centrum für Hämatologie und Onkologie Bethanien Frankfurt; Advisory Role: Beratungstätigkeit Novartis; Financing of Scientific Research: Novartis

Oncol Res Treat 2016;39(suppl 3):1–104

Tesch H.1, Pelz H.2, Janssen J.3, Hansen R.4, Fietz T.5, Anhuf J.6, Haeberle L.7, Belleville E.8, Schardt C.9, Azeh I.9, Steinmetz T.10 Niedergelassener Onkologe im Centrum für Hämatologie und Onkologie Bethanien Frankfurt, Frankfurt, Germany, 2Ambulantes Therapiezentrum für Hämatologie und Onkologie, Offenburg, Germany, 3Gemeinschaftspraxis für Hämatologie und Onkologie, Westerstede, Germany, 4IDGGQ, Institut für med. Dokumentation GbR, Kaiserslautern, Germany, 5Schwerpunktpraxis für Hämatologie und Internistische Onkologie, Singen, Germany, 6Onkologie Duisburg Nord, Duisburg, Germany, 7Comprehensive Cancer Center ErlangenEMN, Erlangen, Germany, 8ClinSol GmbH & Co.KG, Würzburg, Germany, 9Onko. Logix GmbH & Co. KG, Gelsenkirchen, Germany, 10Praxis für Hämatologie und Onkologie, Köln, Germany 1

Introduction: Despite tyrosine kinase inhibitors (TKI) remarkable efficacy in clinical trials, there is a lack of published data on how CML is managed in the real- world clinical practice settings. The non-interventional study (NIS) DasPAQT is designed to document real-world data on Dasatinib treatment of patients with CML in chronic phase in clinical routine. As most CML patients are treated outside of clinical studies focus lies on health care provided in office based physicians. Methods: Patient population consists of subjects with newly diagnosed Ph+ CP-CML and CML patients in chronic phase resistant or intolerant to prior therapies that are treated with Dasatinib according to the clinical routine. Follow up is documented for a maximum of 24 months. A total of 300 patients from up to 75 sites (hematological and oncological hospitals or practices) will be documented in order to answer the following questions: What is the treatment pattern of Dasatinib in CML patients in a real-world environment, including Dasatinib treatment strategies in first-line chronic CML or in a switch setting? What is the effectiveness and outcome of the Dasatinib treatment and which prognostic clinical and scientific factors determine the specific treatment strategy? What is the patient-reported benefit and the impact of first-line Dasatinib CML treatment on patient quality of life? What are the rates of adherence/compliance, how satisfied are patients with their treatment, what patient-related factors lead to treatment discontinuation? Can long-term treatment response be predicted in a real-world setting? Results: Data on diagnostic monitoring, treatment setting and treatment duration with Dasatinib as well as baseline characteristics and safety data of the first 150 patients will be presented. Conclusion: DasPAQT is intended to provide insight into the routine health care management of CML-patients treated with Dasatinib and its related outcomes. The factors that CML patients and treating physicians may encounter in a real-world setting will be observed and analyzed to understand the benefits and effectiveness of Dasatinib CML treatment. Disclosure: Hans Tesch: Employment or Leadership Position: Niedergelassener Onkologe im Centrum für Hämatologie und Onkologie Bethanien Frankfurt.; Advisory Role: Beratungstätigkeit u.a. Novartis, Roche, Bristol-Myers Squibb; Financing of Scientific Research: u.a. Novartis, Roche, Bristol-Myers Squibb; Expert Testimony: Ja, Bristol-Myers Squibb Tilmann Steinmetz: Employment or Leadership Position: Geschäftsführer X-Med GmbH, Leitung: Onkologie-Köln.; Advisory Role: Amgen; BMS, Celgene, Hexal-Sandoz, Medice, Novartis; Janssen-Cilag; Pharmacosmos; Roche; Vifor; Ariad; Stock Ownership: Kommanditist von GermanOncology; Financing of Scientific Research: KV-No, Privatpatienten; Expert Testimony: Untersuchungen Studienleitung IIT/NIS/Register: Amgen, Celgene, Novartis, Vifor; Other Financial Relationships: Reisekosten: Alexion, Amgen, Bayer, Celgene, Janssen-Cilag, Novartis, Sanofi, Vifor; Immaterial Conflict of Interests: Mitgliedschaften: DGHO, ESMO, DGPM, BNHO, NIONo (Vorsitz).

Abstracts

CONTENTS AUTHOR INDEX

P213

P214

R-EFECT: A multicenter, retrospective evaluation of CML therapy in Austria assessing the impact of early treatment response on patient outcomes in a real-life setting

Standard of care of patients with CML treated in community based oncology group practices in Rhineland-Palatinate (Germany)

Petzer A.1, Herndlhofer S.2, Weltermann A.3, Sliwa T.4, Schmidt S.5, Greil R.6, Wölfler A.7, Wiesholzer M.8, Dormann C.1, Thaler J.9, Tinchon C.10, Ruckser R.11, Lang A.12, Hänig J.13, Winiger I.13, Muenchmeier N.13, Sperr W.R.2

Weide R.1, Rendenbach B.2, Grundheber M.3, Burkhard O.4, Behringer J.5, Maasberg M.6, Ehscheidt P.7, Strehl J.W.8, Hansen R.9, Feiten S.10

Barmherzige Schwestern Hospital Linz, Department of Medical Oncology, Hematology and Gastroenterology, Linz, Austria, 2Medical University of Vienna, Department of Internal Medicine I, Division of Hematology, Vienna, Austria, 3 Krankenhaus der Elisabethinen Linz, Department of Medicine I – Hematology with Stem Cell Transplantation, Hemostasis and Medical Oncology, Linz, Austria, 4Hanusch Hospital Vienna, Department of Medicine III: Hematology Oncology, Vienna, Austria, 5University of Innsbruck, Internal Medicine V: Hematology-Oncology, Innsbruck, Austria, 6Paracelsus University Hospital Salzburg, Department of Medicine III, Salzburg, Austria, 7Medical University of Graz, Division of Hematology, Department of Internal Medicine, Graz, Austria, 8University Hospital St. Poelten, Karl Landsteiner University of Health Sciences, Department of Internal Medicine I, St. Poelten, Austria, 9Klinikum Wels-Grieskirchen, Department of Internal Medicine IV, Wels, Austria, 10LKH Leoben-Eisenerz, Department for Hematology and Oncology, Leoben, Austria, 11 Donauspital/ Sozialmedizinisches Zentrum Ost, 2nd Medical Department, Vienna, Austria, 12LKH Feldkirch, Department of Internal Medicine, Feldkirch, Austria, 13Novartis Pharma GmbH, Oncology, Vienna, Austria 1

Introduction: Data from randomized clinical trials indicate that CML patients (pts) reaching early molecular response at 3 months (EMR; < 10% BCR-ABL/ABL) achieve better OS and benefit from a reduced risk of disease progression.This study has aimed to evaluate the impact of an early treatment response (ETR) in daily clinical practice by analyzing patient data available at CML treatment centers in Austria. Methods: 12 participating centers were asked to retrospectively document data of CP-CML pts that were diagnosed between January 2004 and July 2010 and treated frontline with a TKI outside of a clinical trial. A minimum of 5 years of follow-up was required unless an earlier time of death was known. The term “ETR” was defined as EMR (< 10% BCR-ABL/ABL) or optimal cytogenetic response at 3 months according to ELN 2013 criteria (i.e. MCyR; Ph+ ≤35%). Additionally, if neither BCR-ABL/ABL IS values nor cytogenetic response data were available, pts were scored for ETR using raw BCR-ABL/ABL values. This was supported by a subgroup analysis which demonstrated a good comparability between raw and IS values. Results: Of a total of 211 documented pts 35 were excluded due to missing or unevaluable response data at 3 months, resulting in an analysis cohort of 176 pts. Median age at diagnosis was 56 years, with 40.3% being female. ETR was achieved by 77.3% of pts and associated with a higher 5 year OS (92.5%; p = 0.02) and PFS (95.6%; p = 0.06) with no early deaths by 12 months compared to ETR negative (ETR-) pts with an OS of 77.5% and a PFS of 87.5% and 2 early deaths by 12 months, respectively. At the last visit (median follow-up was 94.5 months and similar in both subgroups) differences in OS and PFS were even more pronounced between ETR+ and ETR- pts (OS 88.1% vs. 67.5%, p = 0.003; PFS 92.6% vs. 84.2%, p = 0.055). Interestingly, the majority of pts reaching ETR had low and intermediate SOKAL scores at diagnosis (low/int/high: 52.9%/34.1%/12.9%), whereas pts failing ETR had predominantly intermediate or high SOKAL scores (20.0%/52.0%/28.0%). Pts reaching ETR were less likely to be switched from imatinib to another TKI (76.7% vs. 55.3% still on imatinib at last visit; p = 0.02). Conclusion: These data from a real life setting support the findings from randomized trials demonstrating that ETR is associated with superior PFS and OS. Pts failing ETR should be closely monitored and treated properly according to available guidelines.

Praxisklinik für Hämatologie und Onkologie, Koblenz, Germany, Gemeinschaftspraxis für Hämatologie, Onkologie und Nephrologie, Trier, Germany, 3Onkologische Schwerpunktpraxis, Trier, Germany, 4Internistische Gemeinschaftspraxis Hämatologie, Onkologie, Palliativmedizin, Worms, Germany, 5Onkologische Schwerpunktpraxis, Speyer, Germany, 6 Gemeinschaftspraxis für Hämatologie und Onkologie, Mayen, Germany, 7Praxis für Hämatologie und Onkologie, Neuwied, Germany, 8Schwerpunktpraxis Hämatologie und Internistische Onkologie, Altenkirchen, Germany, 9 Schwerpunktpraxis für Hämatologie und Onkologie, Kaiserslautern, Germany, 10 Institut für Versorgungsforschung in der Onkologie, Koblenz, Germany 1 2

Introduction: Significant progress has been made in CML-therapy since the introduction of imatinib and other tyrosine kinase inhibitors (TKI) into clinical care. The aim of this study was to assess diagnosis, treatment and outcome of CML-patients who received their treatment in community based oncology practices in Rhineland-Palatinate and whether European LeukemiaNET-guidelines were followed. Methods: All Ph-/BCR-ABL-positive CML-patients who were treated between 12/2001-12/2015 in 9 oncology group practices were analysed retrospectively concerning diagnosis, treatment and outcome according to European LeukemiaNET-guidelines. Data were collected from patient files into a central data base and analysed statistically with SPSS. Results: 264 patients (pts) with a median age of 60 (18-90) were analysed. 126 (48%) were female, 138 (52%) were male. At initial diagnosis bone marrow biopsy was performed in 213 pts (81%). Cytogenetics was applied in 204 pts (77%) (38% in blood, 56% in bone marrow). FISH-analysis was used in 155 pts (59%) (33% in blood, 36% in bone marrow). PCR-testing to detect a BCR-ABL1-rearrangement was applied in 200 pts (76%) (52% blood, 37% bone marrow). 258 pts (98%) were in chronic phase, 5 (2%) in accelerated phase and 1 (0.4%) in blast crisis at diagnosis. EUTOS score could be calculated in 131 pts (50%). 20% were high risk, 80% low risk. 252 pts (95%) received some form of TKI-therapy. Out of 416 TKI-therapies 308 (74%) were PCR-based monitored, 148 (36%) were monitored by cytogenetics. First line treatment was imatinib in 201 pts (80%), 51 pts (20%) received a second generation TKI. Second line treatment consisted of dasatinib in 59%, nilotinib in 32%, imatinib in 6% and bosutinib in 3%. Third line treatment was nilotinib in 56%, dasatinib in 35%, ponatinib in 6% and imatinib in 3%. 62 pts (23%) were treated within a study protocol. 13 pts (5%) received an allogeneic transplantation. Overall survival probability was 88% after 5 years and 72% after 10 years. Disease specific survival was 95% after 5 years and 86% after 10 years. Conclusion: The overwhelming majority of CML-patients treated in oncology group practices receive standard of care as suggested by European LeukemiaNET-guidelines. Overall survival in routine care is comparable to international studies. Disclosure: No conflict of interest disclosed.

Disclosure: Andreas Petzer: Advisory Role: Novartis, BMS, Pfizer, Ariad; Financing of Scientific Research: Novartis, BMS, Ariad; Expert Testimony: Novartis Wolfgang Sperr: Financing of Scientific Research: Novartis

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

CONTENTS AUTHOR INDEX

P215

Treatment results of 13 years CML therapy in a private office. A contribution of the German CML Alliance Kämpfe D.1, Haverkamp T.2, Heil G.3, Schulte C.4, Tesch H.5, Stein H.6, Die Deutsche CML-Allianz Praxis für Hämatologie / Onkologie, Lüdenscheid, Germany, 2MVZ Dr. Eberhard & Partner Dortmund (ÜBAG), Dortmund, Germany, 3Klinik für Hämatologie und Onkologie am Klinikum Lüdenscheid – Märkische Gesundheitsholding GmbH und Co KG, Lüdenscheid, Germany, 4Institut für Hämatopathologie, Hamburg, Germany, 5Centrum für Hämatologie und Onkologie Bethanien, Frankfurt / Main, Germany, 6Pathodiagnostik Berlin, Berliner Referenzzentrum für Lymphom- und Hämatopathologie, Berlin, Germany 1

Introduction: Many patients with CML are diagnosed and treated in private offices of hematologists/oncologists in Germany. In registry studies it was found that only about one quarter of these patients are included in the context of clinical studies. Here we present the analysis of a complete, non-selected data set of all patients with CML disease, who have been treated by one doctor in a private office during the last 13 years. Methods: During the period from 07/2003 to 03/2016, all patients diagnosed with CML were identified and their therapy, response and survival were analyzed. The analysis was based on medical reports, laboratory data and prescription data of these patients. Demographics and treatment outcomes of these patients were compared with data from studies (CML IV, IRIS). Results: Altogether 51 patients were treated with CML in this period, 33 patients were newly diagnosed (50 Ph+, 1 Ph- CML). 48 (96%) patients were in the CP, 2 (4%) patients in AP at diagnosis (ID 01/97…03/2016 with median age of 62 (22 ... 88) years). One patient was autologous and another patient was allogeneic transplanted before the evaluation period (4%). 5 (10%) patients had at least one additional neoplasia. During the follow-up period (median: 5.5 (0 ... 20.5) years), two patients had an acceleration during therapy, both reached subsequent CPs under treatment again. A 67 year old woman with ovarian cancer died in a blast crisis. Analysis of 3.785 mo. of therapy split up in 65% IMA±IFN, 12% IFN±HU, 6% NIL±IFN, 6% no treatment (5% TFR, 1% break) and 2% DAS (other therapies remaining). With 12 (24%) patients one or more mutational analysis of the bcr-abl gene were performed. With 3 (6%) patients there were mutations found, one with T315I mutation. With patients with a delayed start therapy with TKI (> 90 after ID) fewer reached a molecular response (eg. MMR 72 vs. 90%, p < 0.05) and they required a significantly longer time to reach these responses (median time to MMR 126 vs. 15 months, p < 0.05). Until now a total of 5 (10%) patients achieved a treatment free remission (TFR) successfully after achieving a deep and stable molecular remission (median TFR 38 (8 ... 98) months). Conclusions: Comparable treatment results can be achieved for patients with CML in a private office as well as in studies with a very similar survival rate. Small differences could be easily explained by different age (older patients in office) and comorbidities (unselected patient cohort). Disclosure: Dietrich Kämpfe: Immaterial Conflict of Interests: logistische Unterstützung durch die Firma ARIAD Pharmaceuticals Inc. Harald Stein: No conflict of interest disclosed.

Posterdiskussion

Myelodysplastisches Syndrom, sonstige Hämatologie P216

Highly variable Separase activity patterns in bone marrow of patients with myelodysplastic syndrome and acute myeloid leukemia Ruppenthal S.1, Prinzhorn W.1, Kleiner H.1, Lammer F.1, Nowak D.1, Hofmann W.-K.1, Fabarius A.1, Seifarth W.1 Department of Hematology and Oncology, University Medical Centre, Mannheim, Germany 1

Introduction: ESPL1/Separase, an endopeptidase, is a key player for centrosome duplication and separation of sister chromatids in anaphase of mitosis. Overexpression and deregulated proteolytic activity of Separase is associated with the occurrence of supernumerary centrosomes, chromosomal missegregation and aneuploidy, as frequently observed in human cancers. Increased Separase proteolytic activity in a small subpopulation of tumor cells may serve as a driver of tumor heterogeneity and clonal evolution in human hematopoietic disorders. Recently, we have shown that in CML patients Separase activity was highly increased when compared to healthy donors. No data are currently available on Separase activity in myelodysplastic syndromes (MDS) and MDS-related secondary acute myeloid leukemia (sAML). Therefore we set out to measure Separase activity in respective clinical bone marrow (BM) samples. Methods: Density gradient centrifugation using Ficoll-Paque was performed to separate mononuclear cells from BM specimen of healthy donors and patients with MDS and AML. Separase activity in living cells was measured by fluorescence-activated cell sorting (FACS) employing a rhodamine 110 (Rh110)-conjugated Rad21 cleavage site peptide as intracellular substrate. The number of Separase-active cells and the intercellular Separase activity distribution (expressed as SAD-value) were calculated for each sample. Results: We have analyzed 33 BM samples derived from patients with MDS (n = 18, median age 67.4 years, range 27-85), with AML (n = 11, median age 61.3 years, range 25-73) and healthy donors (n = 4, median age 57 years, range 26-76). A tendency to increased numbers of Separase-active cells in MDS and AML (5.3 and 6.3 vs. 3.8 in healthy donors) and a higher variation of SAD values within the MDS group (inter-sample distribution 8.4, range 6.6-15.0) and AML group (inter-sample distribution 11.5, range 6.5-18) compared to healthy donors (inter-sample distribution 4.5, range 9.1-13.6) was observed. A significantly higher SAD value in sAML group was detected when compared to primary AML and MDS (sAML vs. MDS, p = 0.0345; sAML vs. AML, p = 0.0124). Conclusion: We observed an increasing occurrence of highly variable Separase activity patterns concurring with malignancy. This high variability may reflect the transformation process from MDS into sAML. To corroborate this hypothesis with respect to clinical data, a higher number of patients and controls will be investigated. Disclosure: No conflict of interest disclosed. P217

Recurrent mutations, expression analysis and functional characterization of cohesin subunits in myelodysplastic syndromes and acute myeloid leukemia Abolfathi M.1, Schroeder T.2, Kartal-Kaess M.1, Bochtler T.1, Roßberg A.1, Jauch A.3, Haas R.2, Krämer A.1 Clinical Cooperation Unit Molecular Hematology/Oncology, German Cancer Research Center (DKFZ) and Department of Internal Medicine V, University of Heidelberg, Heidelberg, Germany, 2Department of Hematology, Oncology and Clinical Immunology, Heinrich-Heine-University, Düsseldorf, Germany, 3Institute of Human Genetics, University of Heidelberg, Heidelberg, Germany 1

Myelodysplastic syndromes (MDS) are hematopoietic stem cell disorders that are associated with bone marrow failure and transformation into acute myeloid leukemia (AML). An increasing list of genes including the

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cohesin complex subunits STAG2, RAD21, SMC1 and SMC3A has been described to be mutated in both MDS and AML. Whether cohesin mutations contribute to leukemogenesis via interference with gene expression or chromosomal stability remains controversial. In this study, we analyzed a panel of 63 genes for mutations in 63 MDS blood or bone marrow samples using a targeted re-sequencing approach. STAG2 was mutated in only 3/63 (4.8%) of samples. No mutations in other cohesin components were found. On the other hand, STAG2 expression was lost in 18 out of 74 (24.3%) AML samples due to STAG2 mutations in 20% (2/10) and promoter methylation in 58.3% (7/12) of cases. In addition, we used CRISPR/Cas9 genome editing to knock out STAG2 in diploid, chromosomally stable HCT116-p53+/+ and HCT116-p53-/- cells. Whereas loss of STAG2 led to alterations in gene expression profiles in both cell lines, chromosome aberrations were only induced in the HCT116-p53-/- background. We conclude that the expression of STAG2 is lost in about one quarter of AML cases, frequently as a consequence of promoter methylation. Depending on the genetic background, both disturbed gene expression and aneuploidy are associated with loss of STAG2. Disclosure: No conflict of interest disclosed. P218

Molecular tracking of somatic mutations in MDS patients with monosomy 7 receiving azacytidine Dierks S.1, Martin R.1, Shumilov E.1, Bacher U.1, Ganster C.1, Shirneshan K.1, Flach J.1, Haase D.1 Universitätsmedizin Göttingen, Hämatologie und Medizinische Onkologie, Göttingen, Germany 1

Introduction: Myelodysplastic syndromes (MDS) are heterogeneous hematopoietic stem cell disorders with clonal cytogenetic anomalies in ~50% of cases. In addition to characteristic cytogenetic aberrations like del(5q) or monosomy 7 (-7), recent studies detected somatic molecular mutations in 80-90% of MDS patients (pts). Demethylating agents e.g. 5-azacytidine (AZA, Vidaza®) are an effective option for high-risk MDS pts, especially if they have a monosomy 7. However, response to AZA is achieved in a part of pts only and remains temporary. Still the molecular mechanisms that underlie response as well as primary/secondary resistance are poorly understood. Methods: Sanger sequencing (comprising 17 recurrently mutated genes) was used to identify mutations in high-risk MDS patients with -7 either as isolated or as combined aberration. Subsequently, peripheral blood samples of 2 patients with an isolated -7 were retrospectively analyzed from time points before, during and after AZA therapy by next generation sequencing (NGS) (sensitivity: ~5-10%). Results: Both patients with a sole -7 were subjected to AZA therapy. Initial mutations were found in ASXL1, RUNX1 and U2AF1 (pt #1) and SRSF2, IDH2 and RUNX1 (pt #2), respectively. Retrospective NGS analyses before, during and after therapy sensitively detected and allowed reconstruction of molecular evolution of subclones disappearing or emerging during or after AZA. During follow-up Patient #2 acquired a transient JAK2mut as well as mutations in NOTCH1 and CUX1 after AZA was stopped. Thus, certain subclones with mutations in genes like RUNX1, which were associated with -7, responded well to AZA and disappeared, whereas other mutations in genes like CUX1 (Tumor suppressor gene on chr. 7q22.1) emerged after AZA discontinuation. Recently, a third pt with -7 has been identified and will likewise be treated and monitored. Conclusion: The progression of certain clones under AZA may be associated with the emergence of new subclones and devolution of previously existing responsive mutations. Sensitive molecular follow-up analyses of these mutation patterns by NGS may improve the understanding and prediction of therapy response and resistance to demethylating agents in pts with -7. Therefore, improved surveillance of the molecular course under therapy using NGS technology may pave the way to more individualized therapeutic strategies for high-risk MDS pts.

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Deferasirox in the routine-treatment of MDS patients with chronic iron overload: interim results from the noninterventional, prospective study EXSEPT Nolte F.1, Schumann C.2, Bueckner U.3, Schmidt B.4, Hebart H.5, Rubanov O.6, Kühn R.-B.7, Kreil S.2, Metzgeroth G.2, Johrs C.8, Albrecht S.8, Boch T.2, Hofmann W.-K.2 St. Hedwig Krankenhaus, Berlin, Germany, 2University Hospital Mannheim, Medical Faculty Mannheim of the University of Heidelberg, Mannheim, Germany, Department of Hematology and Oncology, Mannheim, Germany, 3 Office-based Hematology/Oncology specialist, Bochum, Germany, 4Officebased Hematology/Oncology specialist, Munich, Germany, 5Stauferklinikum Schwaebisch Gmuend, Internal Medicine, Mutlangen, Germany, 6Officebased Hematology/Oncology specialist, Hameln, Germany, 7Office-based Hematology/Oncology specialist, Oldenburg, Germany, 8Novartis Pharma GmbH, Nuremberg, Germany 1

Introduction: Myelodysplastic disorders (MDS) are oligoclonal stem cell disorders characterized by peripheral cytopenias and an increased risk for progression to acute myeloid leukemia. The majority of patients depends on regular red blood cell (RBC) transfusions during the course of their disease putting them at risk for transfusional iron overload (tIO). The iron chelator deferasirox (DFX) is approved for the treatment of tIO. Methods: The non-interventional study EXSEPT included patients (pts) with IO treated with DFX. We performed a subgroup interim analysis in pts with MDS and tIO. DFX was prescribed according to market authorization. Pts were enrolled between 2010 and 2014 with a follow-up of 2 years. Efficacy was calculated by change in in serum ferritin (SF) in pts for whom SF values were available at 24 mo after DFX treatment start (n = 49). Safety was evaluated in pts with at least one follow-up visit after dosing of DFX (n = 280). Results: 280 MDS pts were enrolled with a median age of 74 yrs (2-93 yrs). Most pts were classified as having either low or int-1 risk according to the IPSS Score (n = 167; 60%). At baseline transfusion frequency was 2-4 RBC per mo in most pts (57%). Mean daily starting dose of DFX was 15.6 mg/kg (SD 6.6) and 169 pts (60%) remained on the selected starting dose. Dose adjustments were done in 111 (40%) pts with dose increases reported in 65 pts (23%), mainly due to an initially planned dose escalation strategy (n = 45; 46%). Insufficient responses led to dose increase in 27 pts (28%). Mean SF levels declined from 2093 µg/mL at baseline (SD 1091) to 1751 µg/mL at 24 months (SD 1179), i.e. total mean change of -441 µg/mL (SD 1364). Premature discontinuation occurred in 195 pts (70%) after a mean time of 253 days (SD 177). Reason for discontinuation was death in 59 pts and adverse events (AEs) in 44 pts. AEs were reported in 249 pts; 32% of the AEs occurred within the first 3 mo. Most frequently reported AEs were decrease in renal creatinine clearance (73 pts; 26%), increase in blood creatinine (53 pts; 19%); fatigue (20 pts; 7%), diarrhea (42 pts; 15%) and nausea (28 pts; 10%). Conclusions: Our data confirm the efficacy of DFX in reducing SF. Safety profile and discontinuation rates were as previously observed. While AEs remain a major reason for discontinuation, especially for older patients, dose adjustment, i.e. dose escalation strategies could be considered for improvement of treatment tolerability. Disclosure: Florian Nolte: Advisory Role: Ja; Financing of Scientific Research: Ja; Expert Testimony: Ja W.-K. Hofmann: Advisory Role: Ja; Financing of Scientific Research: Ja; Expert Testimony: Ja

Disclosure: No conflict of interest disclosed.

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The relevance of the international prognostic scoring system (IPSS) for patients with myelodysplasia (MDS) at higher age Steinmetz H.T.1, Wahdat R.1, Haastert B.2, Sauer A.3, Lathan B.4, Lerchenmüller C.5, Tesch H.6, Germing U.7, Schmitz S.1 Outpatient clinic, hematology + oncology, Cologne, Germany, 2mediStatistica, Neuenrade, Germany, 3Outpatient clinic, hematology + oncology, Potsdam, Germany, 4Outpatient clinic, hematology + oncology, Dortmund, Germany, 5 Outpatient clinic, hematology + oncology, Münster, Germany, 6Outpatient clinic, hematology + oncology, Frankfurt a.M., Germany, 7University Hospital, hematology + oncology, Düsseldorf, Germany 1

The IPSS was developed to estimate the prognosis of patients (pts) with MDS in a cohort of 816 pts of academic centers with a median age of 69 years (y). It was an aim of the regular care MDS-registry to describe the age distribution of MDS in regular care and the meaning of the IPSS in various classes of age. Methods: Pts with written informed consent could be included if a bone marrow biopsy has been performed. They were eligible if basic data and the quarterly course of the disease were documented. Statistical analysis: Data were described overall and stratified by age classes. Corresponding global tests were performed (Chisquare, Kruskal-Wallis). Survival was estimated by Kaplan-Meier curves stratified by IPSS and age being compared by a bivariate Cox regression model. Results: Between July 2009 and March 2016 (81 months) 2,118 pts from 90 institutions mainly outpatient practices, were documented and eligible. Median age of 843 (39.8%) women and 1,275 (60.2%) men were 74.9y (min-max: 26.5 - 94.2). The duration of observation, frequencies of IPSS, need for transfusions (Tx) prior to diagnosis (d), and the Charlson comorbidity index (CCI) are given in the table. Tab. 1. Age distribution and MDS parameter

ITEM N (%) Mean month of observation (mean (SD)) Low risk N (%) a)

ALL 2,118 (100)

- 69y 573 (27.1)

25.52 (21.22)

29.13 27.20 (23.80) (21.29)

24.90 (20.26)

20.48 (17.86)

143 (34.5) 165 (39.9) 71 (17.2)

125 (33.9) 132 (35.8) 76 (20.6)

121 (36.5) 127 (38.3) 55 (16.6)

503 (33.8) 569 Int 1 N (%) a) (38.3) 287 Int 2 N (%) a) (19.3) High risk N 128 (%) a) (8.6) TX prior d N 631 (%) b) (32.5) CCI > 0 N 1,171 (%) c) (55.34) Deceased 756 N (%) (35.7)

70 - 74y 75 - 79y 80y + 501 532 512 (23.7) (25.1) (24.2)

114 (30.7) 145 (39.0) 85 (22.9)

p value –

p = 0.448

p = 0.448 “ “

35 (8.5) 28 (7.5) 36 (9.8)

29 (8.7) “

139 (26.3) 274 (47.9) 162 (28.3)

190 (40.9) 320 (62.6) 225 (44.0)

149 (32.7) 247 (49.3) 172 (34.3)

153 (31.0) 330 (62.0) 197 (37.0)

p < 0.0001 p < 0.0001

Fig. 1. Kaplan-Meier curves for IPSS and Age.

Conclusion: Prognostic properties of IPSS seem to be lower in higher age groups, which are more prevalent in the MDS pts in the German regular care registry. So it will be important to look for additional prognostic factors for older pts. Supported by an unrestricted grant from Celgene and Novartis. Disclosure: Hans Steinmetz: Employment or Leadership Position: CEO X-Med GmbH und Onkologie Köln; Advisory Role: Amgen; BMS, Boehringer-Ingelheim, Celgene, Hexal-Sandoz, Medice, Novartis; Janssen-Cilag; Pharmacosmos; Vifor; Stock Ownership: Kommanditist von GermanOncology; Financing of Scientific Research: KV-No, Privatpatienten; Expert Testimony: Studienleitung IIT/NIS: Amgen, Celgene, Novartis, Vifor. Studienarzt für Studien folgender Sponsoren: AIO, Amgen, BMS, Celgene, IoMedico, Janssen-Cilag, Kompetenznetz Maligne Lymphome und KN Akute Leukämien, Lilly, Merck, Novartis, Pfizer, Pharmacosmos; Other Financial Relationships: Reisekosten: Alexion, Amgen, Bayer, BMS, Celgene, Janssen-Cilag, Novartis, Sanofi, Vifor; Immaterial Conflict of Interests: Mitgliedschaften: DGHO, ESMO, DGPM, BNHO, NIONo Stephan Schmitz: Employment or Leadership Position: CEO X-Med GmbH und Onkologie Köln; Advisory Role: Amgen; BMS, Boehringer-Ingelheim, Celgene, MSD, Novartis; Janssen-Cilag; Stock Ownership: Kommanditist von GermanOncology; Financing of Scientific Research: KV-No, Privatpatienten; Expert Testimony: Studienarzt für Studien folgender Sponsoren: AIO, Amgen, BMS, Celgene, IoMedico, Janssen-Cilag, Kompetenznetz Maligne Lymphome und KN Akute Leukämien, Lilly, Merck, Novartis, Pfizer; Other Financial Relationships: Reisekosten: BNHO,; Immaterial Conflict of Interests: Mitgliedschaften: DGHO, ESMO, ASCO, DGPM, BNHO, NIONo.

–

Missing values: a) 631, b) 175, c) 2 pts Higher age and IPSS risk were significantly associated with the risk of death, an additional interaction between age and IPSS risk was significant (p = 0.0198, Cox model).

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Successful management of Thrombotic thrombocytopenic purpura in early pregnancy with maternal and fetal survival Voskova D.1, Greul R.1, Lenger D.1, Hellmich U.2, Siedler D.3, Fridrik M.A.1, Fuchs D.1 Kepler Universitätsklinikum, Klinik für Interne 3 - Schwerpunkt Hämatologie und Onkologie, Linz, Austria, 2Kepler Universitätsklinikum, Klinik für Interne 2 - Schwerpunkt Nephrologie, Endokrinologie/Diabetologie, Rheumatologie, Hepatologie, Linz, Austria, 3Kepler Universitätsklinikum, Klinik für Gynäkologie, Geburtshilfe und gynäkologische Endokrinologie, Linz, Austria 1

Thrombotic thrombocytopenic purpura is a rare disease and usually presents with microangiopathic hemolytic anemia and thrombocytopenia (1). Pregnancy is a known risk factor for a first occurrence of TTP, but

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TTP in early pregnancy is extremely rare. The best course of management is unclear (2). We present the case of a nulliparous, 28-year-old women who presented with microangiopathic hemolytic anemia and severe thrombocytopenia (15 G/l) in the 10th week of gestation. TTP was confirmed by ADAMTS13 levels (< 1%) and detectable antibodies. Bone marrow biopsy was unremarkable and the pregnancy was intact. The patient was treated with plasma exchange and steroids (prednisone 1mg/kg/day). Although she developed an anaphylactic reaction to plasma exchange after three sessions, ADAMTS 13 levels had improved and thrombocytes were normalized. She was continued on prednisone (tapered and discontinued in gestational week 30) and enoxaparine 40mg/ day. She delivered a healthy male infant per caesarean section in the 37+4 week of gestation. There are fewer then 30 published cases of TTP in early pregnancy in the literature. Maternal survival was excellent, but fetal survival remained poor in the majority of published cases. We could show that TTP in early pregnancy can be successfully managed with plasma exchange with fetal and maternal survival. References: 1 Moschcowitz E.: An acute febrile pleiochromic anemia with hyaline thrombosis of the terminal arterioles and capillaries: an undescribed disease. Arch Intern Med. 1925 Jan 1;36(1):89-93. 2 Scully M, Thomas M, Underwood M, Watson H, Langley K, Camilleri RS, et al.: Throm-botic thrombocytopenic purpura and pregnancy: presentation, management, and sub-sequent pregnancy outcomes. Blood. 2014 Jul 10;124(2):211-219. Disclosure: No conflict of interest disclosed. P222

Aortic thrombosis leading to the diagnosis of Paroxysmal Nocturnal Hemoglobinuria (PNH) Alashkar F.1, Schemuth H.2, Herich-Terhürne D.1, Dührsen U.1, Röth A.1 Universitätsklinikum Essen, Westdeutsches Tumorzentrum, Klinik für Hämatologie, Essen, Germany, 2Universitätsklinikum Essen, Institut für Diagnostische und Interventionelle Radiologie und Neuroradiologie, Essen, Germany 1

Introduction: Paroxysmal nocturnal hemoglobinuria (PNH) is a rare clonal blood disorder of the hematopoietic stem cells characterized by chronic, uncontrolled complement-mediated intravascular hemolysis and platelet activation resulting in thrombophilia with thromboembolic events (TEs) and cytopenia due to bone marrow failure. Eculizumab, an anti-C5 monoclonal antibody, has been proven to reduce PNH-related complications, including TEs. Patient / Results: In Sep 2010 a 57-year-old female patient was referred to a general hospital for investigation due to recurrent, unexplainable abdominal painful crises being evident since 2008. Initially, the diagnosis of a perforated sigma diverticulitis with an abdominal abscess was made, and a sigma resection was performed. However, abdominal crises continued by Oct and Dec 2010 and further invasive work-up showed gastritis. Surprisingly, an endoscopic ultrasound of the stomach indentified a partial thrombosis of the thoracic and abdominal aorta (hemoglobin (Hb) 11.4g/dL, platelets (PLTs) 129/nL, lactate dehydrogenase (LDH) 287U/L). The medical history was otherwise unremarkable for TEs, thus, phenprocoumon was initiated and switched to acetylsalicylic acid by Aug 2013, as the aortic thrombus underwent regressive changes in the follow-up CTscan. In Sep 2013 abdominal painful crises relapsed, and even a papillotomy was performed due to the impression of a papillary sclerosis with mild dilation of the ductus choledochus. With the development of a progressive thrombocytopenia (70/nL), the diagnosis of PNH was made by flow-cytometry. The patient was referred to our centre in Dec 2013 and treatment with eculizumab was initiated because of symptomatic PNH (thrombosis, abdominal crises). By the time of presentation, laboratory evaluation revealed a mild hemolytic anemia (11.1g/dL) with an increased LDH (1167U/L), an increase of D-dimer (1mg/L), and a PNH clone size of 63%

Abstracts

(Gran., FLAER). Until now, eculizumab is well tolerated and no further abdominal crises or TEs were observed (Hb 9.9g/dL; PLTs 59/nL; LDH 295U/L; D-dimer 0.28mg/L). Conclusion: In PNH up to 40% of the patients develop TEs and in 21% of the cases, TEs precede the diagnosis of PNH. Abdominal pain, dyspnea or chest pain are possible predictors for TEs. Screening for thrombophilia should include PNH diagnostics by flow-cytometry in patients with thromboembolic complications and evidence of hemolysis, atypical thrombotic locations or unexplained thromboses. Disclosure: Ferras Alashkar: Other Financial Relationships: Reisekostenerstattungen: Alexion Pharamaceuticals Alexander Röth: Advisory Role: Alexion Pharamaceuticals, Novartis, Roche; Financing of Scientific Research: Alexion Pharamaceuticals; Expert Testimony: Alexion Pharamaceuticals, Geron; Other Financial Relationships: Reisekostenerstattungen: Alexion Pharamaceuticals P223

Infection control with granulocyte transfusions in very severe aplastic anemia (VSAA) Farsijani N.M.1, Dührsen U.1, Röth A.1 Uniklinik Essen, Hämatologie, Essen, Germany

Introduction: Aplastic anemia commonly results from auto-reactive T-cell-mediated hematopoietic stem cell (HSC) suppression, mandating intensive immunosuppressive therapy (IST) with cyclosporine A and antithymocyte globulin when HSC transplantation is not feasible. Good IST response rates up to 80% are hampered by fatal infections due to neutropenia and therapy-related immunosuppression. To replenish the neutropenic immune system granulocyte substitution with allogeneic granulocytes has been introduced since the 1960s. However, lack of sufficiently powered clinical trials has rendered this treatment approach controversial and rarely used. Patient/Results: A 72-year-old woman was diagnosed with VSAA and subsequently treated with IST in 10/2014. 2 weeks after IST initiation she was admitted to the hospital with neutropenic fever and a CRP value of 14.7 mg/dl. Broad spectrum anti-infective therapy was initiated but despite the use of several different agents including piperacillin/tazobactam, clarithromycin, meropenem, aciclovir, voriconazole, and liposomal amphotericin B (LAMB), fevers aggravated with rising CRP values up to 33.0 mg/dl. Chest computer tomography (CT) revealed a solitary infiltrate in the left lower pulmonary lobe, highly suspicious of invasive aspergillosis. With lack of other treatment options allogeneic granulocyte transfusions were initiated starting in 12/2014. Additionally to conventional anti-infective therapy with meropenem, clarithromycin, and LAMB, 4 granulocyte transfusions were carried out every other day. Per transfusion the total PMNC number ranged from 11.97*10^9 to 73.61*10^9 with an average PMNC number of 50.88*10^9. This led to a significant increase in PMNC blood counts, and associated with falling CRP values down to 6.9 mg/dl and resolution of fever. 7 days after the last transfusion CRP values rose again without change in conventional anti-infective medication, so that granulocyte transfusions were re-initiated. This led to an immediate response with persistent CRP values < 5 mg/dl and hospital discharge. Control chest CT confirmed resolution of the pulmonary infectious focus. Conclusion: Granulocyte transfusions represent a valuable treatment option in refractory neutropenic infections, warranting reliable randomized trials. Especially in the setting of VSAA where neutrophil recovery after ISH is anticipated granulocyte transfusions should be considered when other anti-infective treatments have failed. Disclosure: Navid Farsijani: No conflict of interest disclosed. Alexander Röth: Advisory Role: Alexion, Novartis; Expert Testimony: Alexion

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Care situation of patients with iron deficiency/iron deficiency anaemia in Germany Hubmann M.1, Maier D.2, Rosmolen J.C.3, Dietzfelbinger H.1 Internistische Praxis für Hämatologie und Onkologie, Herrsching, Germany, emphasis – Institut für Marktforschung im Gesundheitswesen GmbH, München, Germany, 3Vifor Pharma Deutschland GmbH, München, Germany 1 2

Introduction: Iron deficiency (ID) is one of most severe and important nutritional deficiencies worldwide and is thought to be the most common cause of anaemia (approx. 80%). Despite its possible severe clinical sequelae, ID is often underdiagnosed and undertreated. A market research study examined how often practising physicians in Germany diagnose ID/iron deficiency anaemia (IDA), its main symptoms, diagnostics and treatment. Methods: Between 09/2015 and 03/2016, 827 general practitioners and internists were asked in a semi-structured interview how they diagnose and treat ID/IDA. They answered 22 detailed questions based on specific patient data from the previous quarter. Data evaluation was anonymised and performed for Germany as a whole. Results: Patient structure: The respondents treat a mean of 27.9 ID/IDA patients (median: 20; range: 1-400) per quarter. The most common cause of ID/ IDA was cited as hypermenorrhoea (30%), followed by gastroenterological disorders (21%), chronic heart failure (10%), renal failure (10%) and cancers (9%). Symptoms: ID/IDA patients most often report exhaustion (93%), fatigue (92%) and impaired capacity (85%). Diagnostics: Most respondents cited Hb (95%) and serum ferritin values (87%) as standard parameters in ID diagnosis. 40% cited transferrin saturation (TSAT) and 28% C-reactive protein (CRP) as standard diagnostics. Therapy: 91% of asymptomatic ID/IDA patients usually undergo initial oral therapy, whereas ⅓ of symptomatic patients are given i.v. iron. In severe chronic diseases, such as inflammatory bowel disease (IBD) or cancers, i.v. iron is often first-line treatment (56/43%, respectively). Reasons for switching from oral to i.v. therapy include intolerance and insufficient response to oral therapy (95%) or an urgent need for iron (89%). Conclusions: In practice, severe chronic diseases, as well as hypermenorrhoea, are relevant causes of ID/IDA. In primary diseases often accompanied by a chronic inflammatory reaction (IBD, renal failure, heart failure, cancers), ID is treated slightly more often with i.v. iron products. The diagnostics cited are often insufficient to confirm ID (most respondents did not use CRP and TSAT as standard diagnostics). The decision on whether to use oral or i.v. iron replacement was apparently guided more by ID/IDA symptoms than by pathophysiological considerations. Disclosure: Max Hubmann: No conflict of interest disclosed. Hermann Dietzfelbinger: Other Financial Relationships: Sponsoring für Herrschinger Hämato-Onkologie-Symposien P225

c-Cbl regulates c-Mpl receptor trafficking and its internalization Saur S.J.1, Märklin M.1, Ganser M.1, Kanz L.1, Kopp H.-G.1, Müller M.1 Universitätsklinikum Tübingen, Medizinische Klinik und Poliklinik, Hämatologie und internistische Onkologie, Tübingen, Germany 1

Megakaryopoiesis is controlled by a variety of hematopoietic growth factors to maintain physiological levels of circulating platelets. Thrombopoietin (TPO) signalling via its receptor c-Mpl is a key regulator of megakaryopoiesis. Consequently, TPO/c-Mpl signalling needs to be tightly regulated to maintain physiological megakaryopoiesis. One of the most effective mechanisms to permanently disable activated signalling proteins is by targeted degradation via lysosomes or proteasomes. Previous studies have identified c-Cbl as an E3 ligase responsible for the ubiquitination of c-Mpl in cell lines. In this study, we investigated the mechanisms of TPO-mediated c-Mpl degradation in primary mouse cells.

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In order to determine the potential role of c-Cbl in murine megakaryopoiesis we used a conditional PF4-Cre c-Cbl knockout (ko) mouse model to specifically delete c-Cbl in the megakaryocytic lineage. Megakaryocytes were generated in vitro by culturing bone marrow from WT and conditional c-Cbl ko lines for 72 hrs in the presence of rmTPO. Conditional c-Cbl ko mice showed significant bone marrow megakaryocyte hyperplasia. Platelet counts were significantly elevated and platelets were of smaller size in c-Cbl ko mice as compared to control mice. We furthert demonstrate that there were more young platelets produced within a 24 h period in the c-Cbl ko mice although the half-life of platelets was similar in the both cohorts. C-Cbl ko mice showed a severe defect in thrombus formation as assessed in an in vivo thrombus formation model with Fe3Cl. Although TPO plasma levels were increased in the c-Cbl ko mice, there was no difference in liver mRNA levels in the two cohorts. We found that c-Cbl ko mice express more c-Mpl on protein and mRNA levels compared with wild type control. In addition, c-Mpl surface expression was reduced and internalization of the receptor was significantly impaired after TPO stimulation in c-Cbl ko mice. After incubating platelets in vitro with TPO, we found c-Cbl ko platelets to show a severe TPO uptake defect compared with wild type control platelets. Taken together, we demonstrated that c-Cbl ablation leads to a reduced c-Mpl surface expression and an impaired internalization, which results in increased plasma TPO levels culminating in increased megakaryopoiesis. These data enhance our understanding of the regulation of TPO signalling and the physiological role of c-Cbl in the megakaryocytic lineage. Disclosure: No conflict of interest disclosed. P226

Evaluation of the XN Series Bodyfluid Software with special consideration of oncological samples Hughes D.1, Stamminger G.1 Labor Chemnitz, Chemnitz, Germany

Objective: To evaluate the Body fluid software of the Sysmex XN analyser series and its capability in accurately screening samples containing suspected tumor cells Methods: A total of 118 serous body fluid samples were analysed over a period of 4 months on the XN 1000 (Sysmex) analyser using the body fluid software. Samples comprised of 46 ascites (A) and 42 pleural (P) fluid (collected in EDTA tubes) and 30 CSF samples obtained from in-house patients, all of which were analysed within 2 hours of receipt. Manual cell differential, evaluation of cell changes and screening of tumor cells was performed on each sample following cytospin and staining. The ‘high fluorescent’ cell count (HF) and the pre-determined cut off value of ≥100µl by the manufacturer were then compared against microscopic examination. Results: Following microscopic examination 13 samples (P = 8, A = 3, CSF = 2) were manually flagged as having suspected tumor cells, 8 of which were immunohistochemically confirmed by the pathology department as malignant. Five samples including both CSF samples were not examined by the pathologist but were from patients with already confirmed malignancy. Of these 8 samples 4 (50%) had an average HF value of 33µl. A further 2 samples which were above the cut-off were found to only strongly reactive in nature. The remaining two samples which incidentally had very high cell counts (>1500µl) had HF values above the cut-off. Furthermore 9 samples which had a count greater than 100µl but no tumor cells and also examined by the pathologist were found to be reactive in nature. The high HF count was due a high number of benign mesothelial cells which are also highly fluorescent. Conclusion: In high concentrations the body fluid software can be relied upon in screening samples which may contain tumor cells. However at lower counts its capability is questionable. The cut-off of 100µl may allow an unsatisfactory number of samples to slip the net. By reducing the cutoff value to 33µl for example this raises the question of whether the body fluid software leads to any added benefit in screening samples for tumor

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cells. With lowered cut-off values the number of samples which require microscopy would only be marginally reduced. Disclosure: No conflict of interest disclosed.

Posterdiskussion Multiples Myelom 1 P227

Influence of Histone Deacetylase Inhibitors (HDACis) on the expression of adhesion molecules and modern target structures in Multiple Myeloma (MM) Müller S.1, Senger J.2, Waldschmidt J.M.1, Wider D.1, Thomsen A.3, Ihorst G.4, Duyster J.1, Hug M.J.5, Jung M.2, Wäsch R.1, Engelhardt M.1 Universitätsklinikum Freiburg, Department für Hämatologie, Onkologie und Stammzelltransplantation, Freiburg i. Br., Germany, 2Institut für Pharmazeutische Wissenschaften, Alber-Ludwigs-Universität, Freiburg i. Br., Germany, 3Universitätsklinikum Freiburg, Department für Radiologische Diagnostik und Therapie, Freiburg i. Br., Germany, 4Universitätsklinikum Freiburg, Studienzentrum, Freiburg i. Br., Germany, 5Universitätsklinikum Freiburg, Klinikumsapotheke, Freiburg i. Br., Germany 1

Background: During the last decades the vital role of the bone marrow (BM) has increasingly been explored to elucidate the progression and drug resistance in MM, this leading to advances of treatment strategies to modulate the interaction between malignant plasma cells (PCs) and the microenvironment. We here examined the influence of HDACis on the BM niche in a novel 3D co-culture. Methods: The cells are seeded in conical microwells and their proliferation and viability are assessed. These properties are compared both in presence or absence of stroma and 2D cultures. Cells are incubated with HDACis, their synergisms with established antimyeloma drugs and stroma effects upon the treatment can be visualized via flow-cytometry and viability assays. The expression levels of relevant surface molecules (CXCR4, CD38, SLAM-F7, CD138) in primary MM cells and MM cell lines (MMCLs) are determined using FACS-analysis. Results: Inside the microcavities, PCs showed a decreased cell proliferation rate compared to 2D cultures. Cell growth and expression pattern of surface proteins were further influenced by the presence of BM stroma cells (BMSCs), leading to a decrease in CXCR4 but persistence of CD138 levels. Our current results suggest evident differences regarding growth and treatment response in our 3D versus 2D culture systems, which we currently validate using HDACi as a novel substance class in MM. The novel selective HDAC-6 inhibitor JS28 was selected from more than 20 compounds due to trypsin-dependent in vitro HDAC-inhibition assay results. The phenotypic HDACi effect was determined with assessment in 2 MMCLs U266 and RPMI 8226: treatment of MM cells were performed with use of panobinostat or JS28 for 48 hours, either alone or combined with bortezomib, to test synergism. The calculated combination indices (CI) in both MMCLs were < 1 (=synergistic) within a range of approximately EC40 to EC55 for both combined treatment schedules. Conclusions: Our current data underline the importance of accurate 3D co-culture models to mimic in vivo proliferation and drug resistance and to predict later clinical treatment success more effectively. So far, there is insufficient data that conclusively unravel the complex interactions between HDACis, their different subtype selectivity and the increasing number of treatment options in MM. In this context, our ongoing investigations will focus on expression of adhesion molecules, their targeting and their alteration by HDACis.

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Single cell subfractions in the bone marrow vary in their impact on myeloma growth and display potential targets to antagonize MM-stromal protection Waldschmidt J.M.1, Wider D.1, Müller S.1, Follo M.1, Klein C.1, Thomsen A.R.2, Herget G.3, Südkamp N.P.3, Wäsch R.1, Duyster J.1, Engelhardt M.1 Universitätsklinik Freiburg, Dept. Hämatologie, Onkologie und Stammzelltransplantation, Freiburg, Germany, 2Universitätsklinik Freiburg, Dept. Strahlentherapie, Freiburg, Germany, 3Universitätsklinik Freiburg, Dept. Orthopädie und Traumatologie, Freiburg, Germany 1

Introduction: The bone marrow (BM) as a specific tumor microenvironment crucially contributes to quiescence, drug resistance and ultimately to progression of multiple myeloma. However, little is known about the distinct role of niche cell subfractions and their protective impact on myeloma. Our focus here was to more closely study the cellular composition and function of the BM niche by utilizing a novel bone-derived in vitro 3D co-culture platform. Methods: This adhesion-independent three-dimensional co-culture model consisted of an agarose matrix interlayer containing 100 microwells/cm². Each microwell was 1.5mm in depth and permeable for oxygen and cytokines, but not for BMSCs. MMCL and primary BM patient (pt) cells were utilized with and without (w/o) HS-5 vs. M210B4 stromal support. Analyses included trypan blue, Annexin/PI, MTT, FACS, cell cycle analyses and H2B-mCherry/cytochrome c-GFP assays (Udi, BJH 2013). To examine the effect of distinct BM niche cell populations on MM growth, niche cell subsets from C57BL6J mice were acquired, digested and FACS-sorted to collect cell subfractions of mesenchymal stem and progenitor cells (MSPC), endothelial cells, osteoblasts, premature CD146+ MSPCs (PαS) and CXCL12-abundant reticular cells (CaRs). Results: Pt samples after 7 days (d) of culture benefitted from both murine M210-B4 and human HS-5 co-culture. FACS-sorting of murine bone and BM cells led to valid subset separation, illustrated by the multipolar morphology of CD31+ endothelial cells and CD31-, CD45-, Sca1+ MSPCs with fibroblast-like bipolar appearance, whereas PαS cells remained undifferentiated and positive for CD146. We observed that C57BL6J-derived murine cells differed in terms of their growth support for OPM-2 cells after 6d of co-culture: MSPCs from murine BM were more beneficial than those derived from murine bone. BM-MSPCs also induced stronger support than premature MSPCs and CXCL12-abundant reticular cells, commonly considered to be crucial mediators of adhesion in the murine niche. Conclusions: We present a 3D culture model which reflects stromal protection of MM cells and may prove more reliable for ex-vivo drug screening and longitudinal studies. To overcome the limitation of a murine co-culture model, current analyses focus on the generation of human BM cell subsets from both MM pts and control patients suffering from orthopedic diseases. Disclosure: No conflict of interest disclosed.

Disclosure: No conflict of interest disclosed.

Abstracts

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Lenalidomide enhances MOR202 dependent macrophagemediated effector functions via the vitamin D pathway

Long term in vitro survival of plasma cells derived from bone marrow of multiple myeloma patients

Bruns H.1, Busch L.1, Böttcher M.1, Mougiakakos D.1, Bittenbring J.T.2, Nolting J.3, Bisht S.3, Büttner M.4, Rehli M.5, Wimmer J.5, Volmer D.6, Beier F.7, Gezer D.7, Neumann F.2, Bach C.1, Balzer H.1, Moi S.1, Brossart P.3, Mackensen A.1

Waechter M.1, Nogai A.1, Kühnel A.1, Wulf-Goldenberg A.2, Kunitz A.1, Blau O.1, Schmidt-Hieber M.3, Pezzutto A.1, Jehn C.1, Vuong L.1, Doerken B.1, Blau I.W.1

Medizinische Klinik 5, Universitätsklinikum Erlangen-Nürnberg, Erlangen, Germany, 2Medizinische Klinik 1, Saarland University Medical School, Homburg, Germany, 3Medizinische Klinik III, Klinik für Hämatologie, Onkologie und Rheumatologie, Bonn, Germany, 4Institut für Pathologie, Erlangen, Germany, 5 Department of Internal Medicine III, Hematology and Oncology, University Hospital of Regensburg, Regensburg, Germany, 6Institute of Bioanalytical Chemistry, Saarland University, Saarbrücken, Germany, 7Department of Oncology, Hematology and Stem Cell Transplantation, RWTH Medical School, Aachen, Germany 1

The bone marrow niche plays a critical role in determining the fate of malignant plasma cells in multiple myeloma (MM). Macrophages are an abundant component of the stromal cell compartment and are believed to support survival and drug resistance of MM cells. Conversely, macrophages are key immune effector cells for the therapeutic effect of monoclonal antibodies. However, myeloma-associated macrophages (MAMs) regularly fail to exert direct effector functions. Lenalidomide, an immunomodulatory agent that enhances antibody dependent cell mediated cytotoxicity (ADCC), has the potential to synergize with MOR202, an anti-CD38 monoclonal IgG1 antibody currently in phase I/IIa for the treatment of MM. Furthermore, vitamin D plays a key role in regulating effector functions of human macrophages. This is closely linked to the expression of the vitamin D-1-hydroxylase CYP27B1, which catalyzes the conversion of 25-hydroxy-vitamin D (25D) to the bioactive form 1,25-di-hydroxy-vitamin D (1,25D). We have previously shown, that vitamin D promotes tumoricidal activity of macrophages and improves the efficacy of rituximab-dependent cytotoxicity. Therefore, we hypothesized that the combination of MOR202 with lenalidomide and MOR202 with 1,25D would enhance the MOR202- dependent macrophage-mediated effector functions against MM cells. Here we report that MAMs exhibit an altered vitamin D metabolism with a reduced expression of the vitamin D receptor (VDR) and CYP27B1. As a consequence MAMs cannot convert 25D into bioactive 1,25D. Given the importance of the vitamin D pathway for antibody mediated cytotoxicity, we screened several drugs for their ability to restore the vitamin D pathway in human macrophages. We found, by RNA-sequencing, that lenalidomide treatment modulates the phenotype of macrophages and isolated MAMs, and that lenalidomide significantly increases the expression of the VDR and CYP27B1. Furthermore, we demonstrate that isolated MAMs regularly fail to eliminate primary MM cells, and that the lack of effector functions can be overcome by treatment with lenalidomide and vitamin D. Moreover, we show that MOR202-dependent elimination of MM cells is enhanced by pre-treatment of isolated MAMs with lenalidomide and supplementation of vitamin D. In summary, these data show that vitamin D is essential for the effector functions of MAMs and that the therapeutic activation of the vitamin D pathway by lenalidomide may restore their tumoricidal effector mechanisms. Disclosure: Heiko Bruns: Expert Testimony: Finanzierung des Projektes von Morphosys und Celgene Andreas Mackensen: No conflict of interest disclosed.

Charité University Berlin, Department of Hematology, Oncology and Tumorimmunology, Berlin, Germany, 2Berlin-Buch GmbH, Experimental Pharmacology & Oncology, Berlin, Germany, 3Helios Kliniken Berlin, Hematology, Berlin, Germany 1

Introduction: Multiple Myeloma (MM) is a neoplastic B-cell malignancy strongly regulated by bone marrow microenvironment. In vitro co-culture models or in vivo systems using immune deficient mice are normally used for clarification of specific interactions of MM-cells and BMSC and to identify new therapeutic targets. Unfortunately, mesenchymal stem cells are a very rare cell population in bone marrow aspirates (< 0,01%) and their expansion can take several weeks of in vitro culture. MM cells on the other hand are more frequent but don’t survive in culture, separated from their supportive microenvironment. The purpose of our study was to establish a culture system capable of keeping primary MM cells alive over a period of several weeks and thus making it possible to use them together with the expanded MSCs in co-culture or animal trials. Methods: Mononuclear cells (MNCs) were separated from bone marrow aspirates by Ficoll density gradient centrifugation and characterized by FACS analysis for detecting the size of the MM cell population (CD45-/ CD138+). Weekly culture monitoring was done by cell counting using hemocytometer, FACS analysis and Trypan blue staining. After 4 or 5 weeks of culture, cytospins were performed and staining with the Hemacolor staining kit (MERCK) followed by cytomorphological inspection. The monoclonality of the MM cell population was proved by detecting intracellular light chain expression using Simultest anti Kappa/ anti Lambda (BD). The vitality of the MM cells was tested by Annexin staining using the Annexin V-FITC Kit (miltenyi Biotec) as recommended by the manufacturer. Results: Long term cultures of bone marrow MNCs derived from 8 Patients with MM were established and maintained for several weeks. The size of MM cell population in bone marrow aspirates of patients varied between below 10% and more than 50% of the MNCs. Proportion of dead cells (trypan blue positive cells) sustained during the culture period at about 20%. At the end of the culture up to 85% of the MM cells presented as alive in the Annexin test. Both the expression of CD138+ and light chain type were held steady. Conclusions: MM cells are able to survive in vitro together with all their accessory cell types present in the MNC fraction of bone marrow aspirate over a period of several weeks. This makes it possible to apply MSCs and MM cells from the same donor simultaneously for co-culture or animal trials. Disclosure: Marlies Waechter: No conflict of interest disclosed. Igor Wolfgang Blau: Expert Testimony: Grant Celgene P231

RalA and RalB are RAS-independent targets in multiple myeloma cells Seibold M.1, Stühmer T.1, Schmiedl N.1, Mottok A.2, Rosenwald A.2, Chatterjee M.1, Einsele H.1, Bargou R.C.3, Steinbrunn T.1 Universitätsklinikum Würzburg, Medizinische Klinik und Poliklinik II, Würzburg, Germany, 2Universität Würzburg, Pathologisches Institut, Würzburg, Germany, 3 Comprehensive Cancer Center Mainfranken, Würzburg, Germany 1

Introduction: Ral has been branded as a putative effector pathway downstream of oncogenic RAS in several cancer entities, and may promote proliferation, survival and drug resistance of multiple myeloma (MM) cells.

Oncol Res Treat 2016;39(suppl 3):1–104

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We used shRNA-mediated knockdown of RalA and RalB isoforms to appraise their role as potential therapeutic targets. Methods: First, we performed immunohistochemical staining of primary bone marrow trephines of MM patients and Western blotting in MM cell lines to evaluate Ral protein expression. Next, knockdown of RalA or RalB was achieved with transient or stable transfection of MM cell lines by electroporation and the effect on cell survival and apoptosis was measured with flow cytometry using annexin V-APC/propidium iodide staining. To test potential dependence of Ral on oncogenic KRAS or NRAS, Ral pulldown assays were applied. Results: Whereas RalA was prominently expressed in the majority of primary MM cells and MM cells lines, RalB showed modest and heterogeneous expression levels. No obvious correlation with oncogenic RAS mutations could be observed. Abrogation of RalA by shRNA-mediated knockdown impaired MM cell survival in two thirds of the tested cell lines, whereas depletion of RalB did not induce relevant levels of apoptosis in the majority of cells. Surprisingly, Ral activity was proven to be independent of oncogenic KRAS or NRAS mutations at positions 12 or 61. Conclusion: Ral may provide a potential therapeutic target in multiple myeloma independent of the presence of oncogenic RAS mutations. Disclosure: No conflict of interest disclosed. P232

Cyclin D1 C.870G>A polymorphism in patients with multiple myeloma, impact of donor Cyclin D1 C.870G>A polymorphism on the outcome after transplantation Puckert F.M.1, Blau I.W.1, Kühnel A.1, Nogai A.1, Rieger K.1, Kunitz A.1, Hemmati P.1, Arnold R.1, Pezzutto A.1, Doerken B.1, Blau O.1 Charité University Berlin, Department of Hematology, Oncology and Tumorimmunology, Berlin, Germany 1

Background: Deregulation of cyclin D-group is considered an important factor in the pathogenesis of a large number of cancers. Since overexpression of cyclin D disturbs the G1/S transition in the cell cycle, it is considered one of the early key molecular events of cancerogenesis. Previously published data shown a relationship between cyclin D1 (CCND1) c.870G>A polymorphism and risk of t(11;14) in patients with multiple myeloma (MM). Little is known about the prognostic impact of CCND1 c.870G>A polymorphism in MM patients. Moreover, there is no evidence about prognostic impact of the donor CCND1 c.870G>A polymorphism on the outcome after allogeneic stem transplantation (alloSCT) in MM patients. Methods: Peripheral blood samples from 250 MM patients were analyzed. Age of patients ranged from 58 to 85 (at median, 57). In addition, samples from 75 pairs MM patients and stem cells donors were studied. To identify CCND1 c.870G>A polymorphism, PCR assay with endonuclease restrictions was developed. Results: In all MM patients, CCND1 c.870G>A polymorphism was strongly associated with the t(11;14)(q13;q32) (P < 0.001). In MM patients after alloSCT, c.870G-genotype in donors cells was statistically correlated with poor outcome after transplantation (P < 0.01). Moreover, only patients transplanted with “G-Genotype” donors developed secondary cancer after AlloSCT (P < 0.01). No secondary tumors were diagnosed in the group of MM patients received grafts from “A-, AG-genotype” donors. Conclusions: Our results suggest the published data that constitutive genetic factor (CCND1 c.870 G>A polymorphism) is associated with a specific chromosomal translocation in patients with MM. Moreover, we could propose significant impact of genetic polymorphism in donor cells on the outcome after stem cell transplantation. Disclosure: No conflict of interest disclosed.

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Analysis of metabolic factors in modulation of interaction between plasma cells and mesenchymal stromal cells in multiple myeloma patients in vitro Kühnel A.1, Waechter M.1, Kunitz A.1, Nogai A.1, Blau O.1, Pezzutto A.1, Doerken B.1, Blau I.W.1 Charité University Berlin, Department of Hematology, Oncology and Tumorimmunology, Berlin, Germany 1

Introduction: Within the progression and proliferation of multiple myeloma (MM), bone marrow mesenchymal stromal cells (BMMSCs) represent a crucial component in creating a tumor supportive MM microenvironment. Regarding the increasing interest in cancer-metabolism, proliferating cancer cells demand elevated nutrient supply, resulting in an anabolic metabolism described as ‘Warburg effect’. Various GLUT transporters as well as the proliferator-activated receptor-γ coactivator 1α (PGC-1α) and the central growth pathway PI3K/AKT/mTOR are upregulated in MM, contributing to an anabolic state. In the present study, we investigated the role of BMMSCs regarding the Warburg effect by identifying metabolic key molecules in a co-culture system. Methods: BMMSCs from patients (MM-BMMSCs, n = 25) and donors (HD-BMMSCs, n = 5) were isolated and co-cultured with KMS12-PE and JJN-3 cells. Protein levels of GLUT1, GLUT4, PGC-1α and PI3K of mono-and co-cultured cells were detected via western blotting. Pyruvate-kinase activity and Lactate-dehydrogenase activity were measured using customary kits (Pierce™). Results: We have found the increasing of GLUT4 expression in the co-culture of KMS12-PE with MM-BMMSCs compared to mono cell culture. However the expression GLUT4 in JJN-3 showed no significant differences between mono and co-culture. Protein expression of GLUT1 was increased in co-cultures of KMS12-PE and JJN-3 as compared with mono cell culture. Conclusion: Our data suggest that BMMSCs from MM patients can modify expression level of metabolic factors in cell line. This in vitro study confirms the hypothesis that the hematopoietic microenvironment supporting the survival and proliferation of MM cells. Disclosure: Aline Kühnel: No conflict of interest disclosed. Igor Wolfgang Blau: Expert Testimony: Grant Celgene P234

Potent in vitro and in vivo effects of polyclonal anti-humanmyeloma globulins Schieferdecker A.1,2, Shoshani O.3,4, Westner B.5,6, Zipori D.3, Fehse B.1, Kröger N.1, Ayuk F.1 Department of Stem Cell Transplantation, University Medical Center HamburgEppendorf, Hamburg, Germany, 2Department of Oncology and Hematology with section Pneumology, Hubertus Wald Tumorzentrum / UCCH, University Medical Center Hamburg-Eppendorf, Hamburg, Germany, 3Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel, 4San Diego Branch, Ludwig Institute for Cancer Research, La Jolla, United States, 5 Neovii (formerly Fresenius) Biotech GmbH, Gräfelfing, Germany, 6Acino AG, Miesbach, Germany 1

Introduction: Polyclonal anti T lymphocyte globulins (ATG) are used in allogeneic stem cell transplantation to prevent graft versus host disease and have been reported to kill human myeloma cells in vitro and in vivo. We reasoned that polyclonal anti-human-myeloma globulins (AMG) might improve antimyeloma effects without increasing toxicity. Methods: AMG were produced by immunizing rabbits with human myeloma cell lines RPMI-8226 (AMG-8226) or KMS-12-BM (AMG-12-BM). In vitro complement-dependent and -independent cytotoxicity of ATG and AMG were compared in myeloma cells (RPMI-8226, KMS-12-BM, OPM-2), primary T cells and non-hematopoietic cells (Hacat and Panc1). The cytotoxicity assays were analysed by flow cytometry after staining with 7AAD. Combination effects of the polyclonals with bortezomib or melphalan were analysed using Calcusyn®. For analyzing the binding ca-

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pacity of hematopoetic specific antibodies of ATG and AMG an absorption assay was used. In vivo effects were studied in a xenograft NOD-SCID mouse model. Therefore mice were subcutaneously injected with MM1S myeloma cells and thereafter treated with ATG, AMG-8226 or AMG-12BM. Results: Both AMG demonstrated stronger cytotoxicity against myeloma cells compared to ATG. In primary T cells AMG-8226 exerted greater complement-dependent cytotoxicity than ATG, whereas complement-independent cytotoxicity did not differ. Effects on non-hematopoietic cell lines were also similar. Competitive blocking assays revealed four fold more antibodies against CD38 in AMG-8226 compared to ATG. Low concentrations of AMG-8226 and ATG enhanced ADCC. At higher concentrations, ATG had stronger inhibitory effects on ADCC compared to AMG-8226. Combinations of ATG and AMG-8226 with melphalan or bortezomib showed additive to synergistic cytotoxic effects on myeloma cells. In line with in vitro data, mouse experiments with the MM.1S myeloma cell line showed stronger antitumor effects for both AMGs compared to ATG. Conclusion: Our data show more potent antimyeloma effects of AMG compared to ATG with no increase in toxicity and may lay the ground for the development of polyclonal antibodies for the treatment of multiple myeloma. Disclosure: Aneta Schieferdecker: No conflict of interest disclosed. Francis Ayuk: Financing of Scientific Research: Neovii (formerly Fresenius) Biotech GmbH P235

MOR202 alone and in combination with pomalidomide or lenalidomide in relapsed or refractory multiple myeloma: data from clinically relevant cohorts of a phase I/IIa study Raab M.S.1, Chatterjee M.2, Goldschmidt H.1, Agis H.3, Blau I.W.4, Einsele H.2, Engelhardt M.5, Ferstl B.6, Gramatzki M.7, Röllig C.8, Weisel K.9, Jarutat T.10, Weinelt D.10, Boxhammer R.10, Winderlich M.10, Peschel C.11 University Hospital Heidelberg, Department of Medicine V, Heidelberg, Germany, 2University Hospital of Wuerzburg, Department of Internal Medicine II, Wuerzburg, Germany, 3University Hospital of Internal Medicine – AKH Wien, Department of Medicine I, Vienna, Austria, 4Charité Campus Benjamin Franklin, Department of Internal Medicine III, Berlin, Germany, 5Medical University Hospital, Hematology & Oncology Department, Freiburg, Germany, 6 Friedrich-Alexander-University Erlangen-Nuremberg, Department of Internal Medicine 5 - Hematology and Oncology, Erlangen, Germany, 7University Hospital Schleswig-Holstein Division of Stem Cell Transplantation and Immunotherapy, Department of Medicine, Kiel, Germany, 8University Hospital Carl Gustav Carus, Department of Medicine I, Dresden, Germany, 9University Hospital of Tuebingen, Department of Hematology, Oncology, Immunology, Rheumatology and Pulmonology, Tuebingen, Germany, 10MorphoSys AG, Martinsried, Germany, 11Technical University of Munich, Department of Internal Medicine III, Munich, Germany 1

Introduction: MOR202, a HuCAL-derived, human IgG1 CD38 monoclonal antibody, induces potent antibody-dependent cellular cytotoxicity and antibody-dependent cellular phagocytosis. Preclinical models show high activity of single-agent MOR202 and synergy in combination with immunomodulatory drugs (IMiDs), lenalidomide (LEN) or pomalidomide (POM). Unlike other CD38 antibodies, MOR202 does not induce complement-dependent cytotoxicity, thought to be a major contributor to infusion-related reactions (IRRs). The primary objectives of this study were to evaluate the safety, maximum tolerated dose (MTD) and recommended phase II dose of MOR202 in patients with relapsed or refractory multiple myeloma (R-R MM). Methods: This is an interim analysis of a multicenter, dose-escalation phase I/IIa study of MOR202. Preliminary safety and efficacy data from 3 cohorts of patients treated with MOR202 alone or with an IMiD are presented: MOR202 4, 8 and 16 mg/kg weekly; MOR202 8 or 16 mg/kg weekly with either LEN or POM. All patients in these cohorts received prophylactic low dose dexamethasone. Results: As of January 29, 2016, 20 patients had been treated: 11 with MOR202 alone, 5 with MOR202+LEN and 4 with MOR202+POM. Pa-

Oncol Res Treat 2016;39(suppl 3):1–104

tients receiving MOR202 alone or with POM were relapsed or refractory to prior bortezomib and LEN with a median of 4 prior regimens. Patients given MOR202 with LEN had a median of 2 prior regimens, mainly bortezomib, cyclophosphamide and autologous stem cell transplant. The MTD has not been reached. MOR202 alone or with an IMiD was well tolerated with mainly hematological toxicity. No MOR202-related study discontinuations or deaths were recorded. A 2-hour MOR202 infusion was feasible in all patients. Only 1/20 patients had an IRR (grade 1). Responses were reported in 8/18 evaluable patients: 3/10 patients (1 very good partial response [VGPR] and 2 partial responses [PR]) in the MOR202 alone cohort, 3/4 patients (all PR) in the MOR202+LEN cohort and 2/4 patients (1 complete response and 1 VGPR) in the MOR202+POM cohort. At the time of analysis, 7/8 responses were ongoing, with the longest duration >10 months in a patient given MOR202 alone. Preservation of high CD38 levels on MM cells under MOR202 therapy was shown. Conclusions: In this analysis, MOR202 given as a 2-hour infusion showed excellent infusion tolerability and overall safety profile. Promising preliminary efficacy and long-lasting tumor control was seen for MOR202 +/IMiDs in patients with R-R MM. Disclosure: Marc Raab: Advisory Role: Novartis; Amgen; Celgene; MorphoSys; Financing of Scientific Research: Novartis; Amgen; Celgene; MorphoSys; Expert Testimony: Novartis; MorphoSys; Other Financial Relationships: Travel, Accomodation, Expenses: Novartis; Amgen; Celgene; MorphoSys Christian Peschel: Advisory Role: Bristol-Myers Squibb; Financing of Scientific Research: Bristol-Myers Squibb; Amgen; MorphoSys; Other Financial Relationships: Travel, Accomodation, Expenses: Bristol-Myers Squibb; Amgen P236

High-dose therapy (HDT) with melphalan and autologous stem cell transplantation (ASCT) for patient older than 70 years with multiple myeloma – feasible therapy or significant risk of complications? Brockhoff H.1, Meyer zum Büschenfelde C.2, Salwender H.3 Asklepios Campus Hamburg, Hamburg, Germany, 2Asklepios Klinik Altona, Hämatologie/ Onkologie, Hamburg, Germany, 3Asklepios Klinik Altona, Hamburg, Germany 1

Introduction: HDT-ASCT is the standard treatment for patients with multiple myeloma younger than 65. The treatment numbers of older patients (>70 years) with HDT-ASCT grew steadily in the last years. However, there is only a small number of studies yielding insufficient data regarding overall-survival (OS) and risk assessment for these patients. Methods: We retrospectively analyzed a group of 62 patients – median age at ASCT: 71.3 years – with multiple myeloma who underwent HDTASCT treatment at the Asklepios Klinik Altona Hospital between 2004 and 2013. The primary aim was to explore OS and treatment related mortality (TRM). Furthermore, we analyzed potential risk factors such as melphalan-dosage, chronic renal failure (Durie-Salmon stage A/B), tandem-therapy, use of new drugs in induction therapy and response after induction therapy. For survival analysis we used the Kaplan-Meier method. Results: TRM was 0%. The median OS was 51.9 months. Patients who received 200mg/m² melphalan at least once had a longer median OS compared to patients who received a lower dosage (85.3 v. 41.7 months; p = 0.01). Chronic renal failure significantly limited OS (33.7 v. 57.8 months; p = 0.007). Tandem therapy (single: 43.5 v. tandem: 55.9 months; p = 0.39), induction therapy with new drugs (new drugs: 73.9 v. no-new drugs: 55.9 months; p = 0.32) and response after induction therapy (complete remission/ very good partial remission: 73.9 v. partial remission/ minimal remission/stable disease: 49.1 months; p = 0.23) could not be identified as significant risk factors in our analysis. Conclusion: 0% TRM in our patient group shows that older individuals generally tolerate the toxicity of HDT with melphalan well. When patients older than 70 years, without chronic renal failure, are treated with HDT-ASCT, they should receive a 200mg/m² melphalan-regime to achieve better OS. The median OS of 51.9 months is comparable with the standard treatment for multiple myeloma for older patients (>65 years) e.g. Revilimid+ Dexamethasone (RD) or Velcade+ Melphalan+ Predniso-

Abstracts

CONTENTS AUTHOR INDEX

lone (VMP). However, as the administration time for HDT-ASCT is much shorter than RD/ VMP, adverse medical effects of long time glucocorticoid treatment should be considered when choosing a therapy. A dual approach with first-line HDCT-ASCT and RD/VMP in refractory myeloma should be explored in future studies for patients >65years. This could lead to a more economical treatment with a higher quality of life for patients. Disclosure: No conflict of interest disclosed. P237

Standard Operating Procedures (SOPs) of centers of excellence for the diagnosis, treatment and follow-up of multiple myeloma patients: similarities and differences Scheid C.1, Engelhardt M.2, Goldschmidt H.3, Einsele H.4, Starbatty B.5, Bischoff M.6, Glossmann J.-P.7, Skoetz N.5, Centers of excellence working group SOP Universität zu Köln, Klinik I für Innere Medizin, Köln, Germany, 2Uniklinik Freiburg, Klinik I für Innere Medizin, Freiburg, Germany, 3Universitätsklinikum Heidelberg, Hämatologie, Onkologie, Rheumatologie, Heidelberg, Germany, 4Universitätsklinik Würzburg, Medizinische Klinik und Poliklink II, Würzburg, Germany, 5Centrum für Integrierte Onkologie (CIO) KölnBonn, Koordinationsstelle der AG SOP, Köln, Germany, 6Uniklinik Freiburg, Institut für Medizinische Biometrie und Informatik, Freiburg, Germany, 7Centrum für Integrierte Onkologie (CIO) KölnBonn, Köln, Germany 1

Introduction: Currently 13 centers of excellence exist, all funded by the German Cancer Aid (DKH). It is mandatory for them to provide up-todate standard operating procedures (SOPs). Usually, SOPs are developed in one center and consist of clinical pathways which reflect recommendations given by current evidence-based guidelines. In addition, latest research results from clinical trials and specific in-house features are integrated to adapt the recommendations on center-specific needs. If questions of interest may not be covered by guidelines, interdisciplinary experts from the center develop a new SOP. To harmonize methods and evidence processing, the working group SOP has developed a common standard for SOPs (SOP handbook ccc-netzwerk.de). The objective of this project is to describe the process of developing one common network SOP for multiple myeloma. Methods: Four centers (Freiburg, Heidelberg, Köln/Bonn, Würzburg) with high expertise in treatment of myeloma patients indicated interest in developing the network SOP and provided center-specific SOPs. The co-ordinating office of the working group SOP (funded by the DKH, No. 111493) collated all suggestions and recommendations in one clinical pathway to be integrated in the network SOP, in line with the SOP handbook. During that process, a number similarities and discrepancies both in format and content were identified. Results: As the four SOPs differ in their extent and deepness, recommendations are given in various levels of detail. Most of the SOPs describe quite similar diagnostic procedures for myeloma, local standards exist for the usage of computer tomography or MRT. All the SOPs recommend to treat the patient within a clinical trial, latest at relapsed stage. As some of the SOPs directly link to a clinical study register, this information is more up-to-date compared to specific mentioned trials that might be closed before the SOP is updated. As the updating process varies across these SOPs, some also include drugs which were recently approved, as other don’t. The network SOP will be finalized soon and be freely available as a clinical pathway on the CCC network website. Conclusions: The collation of the suggestions and recommendations from four highly experienced centers is a first step in identifying nationwide standards and differences. Harmonization of these recommendations for the network SOP will identify evidence-base gaps and will lead to a standard in Germany for improved myeloma treatment.

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Systematic classification of death causes in multiple myeloma patients Haas E.-M.1, Löpprich M.2, Lücke S.3, Kunz C.3, Pritsch M.1, KnaupGregori P.2, Hillengass J.1, Goldschmidt H.1,4, Mai E.K.1 Department of Hematology and Oncology, University Hospital of Heidelberg, Heidelberg, Germany, 2Institute of Medical Biometry and Informatics, University of Heidelberg, Heidelberg, Germany, 3Division of Biostatistics, German Cancer Research Center, Heidelberg, Germany, 4National Center for Tumor Diseases (NCT), Heidelberg, Germany 1

Introduction: Causes of death (COD) in multiple myeloma (MM) patients and their relation to MM or therapeutic side effects have not been systematically evaluated yet. Methods: A hierarchically structured COD classification was built according to the Qualitative Content Analysis. A superordinate system of categories distinguished (1) MM-dependent, (2) MM-independent, (3) not attributable to (1)/(2) and (4) unknown COD. The MM-dependent COD were further subdivided into (1A) MM progression-related, (1B) therapy-related and (1C) not attributable to (1A)/(1B). A subordinate system defined COD on four levels of different specificity applying the MedDRA terminology (Figure 1).

Fig. 1. Structure of the COD classification.

Disclosure: Christof Scheid: No conflict of interest disclosed. Nicole Skoetz: Employment or Leadership Position: Wissenschaftliche Koordinatorin AG SOP des Netzwerks onkologischer Spitzenzentren

COD of 483 deceased patients, from a cohort of 818 MM patients who had received upfront high-dose therapy, were assessed with the constructed classification. Competing-risks analyses were conducted subsequently on the whole cohort. Results: 80.8% of the COD (n = 483) were MM-dependent, 1.7% MM-independent, 7.0% not attributable to (1)/(2) and 10.6% unknown. Among the MM-dependent COD, 41.8% were MM progression-related, 19.5% therapy-related and 38.7% not attributable to (1A)/(1B). The most common therapy-related COD were sepsis (27.6%), pulmonary sepsis (15.8%) and pneumonia (11.8%). In competing-risks analyses known adverse prognostic factors were associated with an increased risk for MM progression-related death. In contrast, multivariate competing-risks analyses demonstrated that low platelet counts (LPC, < 150/nl; HR = 2.87, p = 0.01) and renal impairment (RI, serum creatinine ≥ 2mg/dl; HR = 2.13, p = 0.11) are associated with an increased risk for therapy-related death. Conclusions: The majority of MM patients died MM-dependent, and among those cases one fifth were related to therapeutic side effects. LPC and RI constitute important risk factors for therapy-related death, reflecting an impaired ability to tolerate MM therapy. Our classification has proved to be reliable and its application in future registries and prospective trials might ensure a reliable collection of data on COD in MM patients.

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

CONTENTS AUTHOR INDEX

Disclosure: Eva-Maria Haas: Other Financial Relationships: Reisekostenerstattung: Bristol-Myers Squibb Elias Mai: Other Financial Relationships: Reisekostenerstattung: Janssen-Cilag, Celgene, Onyx und Mundipharma P239

Bortezomib (BOR)-Thalidomide-Dexamethasone (VTD) as induction treatment for newly diagnosed Multiple Myeloma (MM) is associated with a lower rate of Second Primary Malignancies (SPMs) compared to ThalidomideDexamethasone (TD) Brioli A.1,2, Pezzi A.2, Mügge L.-O.1, Derudas D.3, Petti M.C.4, Zannetti B.A.2, Ferrara F.5, Rocchi S.2, Nobile F.6, Baraldi A.7, Musto P.8, Lanza F.9, Mancuso K.2, Canepa L.10, Catalano L.5, Lazzaro A.11, Pinotti G.12, Boccadoro M.13, Hochhaus A.1, Cavo M.2 Universitätsklinikum Jena, Klinik für Innere Medizin II, Hämatologie und Internistische Onkologie, Jena, Germany, 2Bologna University School of Medicine, Seragnoli Institute of Hematology, Bologna, Italy, 3GIMEMA, Italian Myeloma Network, Cagliari, Italy, 4GIMEMA, Italian Myeloma Network, Roma, Italy, 5GIMEMA, Italian Myeloma Network, Napoli, Italy, 6GIMEMA, Italian Myeloma Network, Reggio Calabria, Italy, 7GIMEMA, Italian Myeloma Network, Alessandria, Italy, 8GIMEMA, Italian Myeloma Network, Rionero in Vulture, Italy, 9 GIMEMA, Italian Myeloma Network, Cremona, Italy, 10GIMEMA, Italian Myeloma Network, Genova, Italy, 11GIMEMA, Italian Myeloma Network, Piacenza, Italy, 12 GIMEMA, Italian Myeloma Network, Varese, Italy, 13GIMEMA, Italian Myeloma Network, Torino, Italy 1

Introduction: The availability immunomodulatory drugs, IMiDs, and proteasome inhibitors, PI, for the front line treatment of Multiple Myeloma (MM) has significantly improved patients’ outcomes, however concerns have been raised regarding the possibility that IMiDs might increase the risk of developing SPMs. Conversely the use of PI in elderly newly diagnosed MM (NDMM) patients (pts) was not associated with an increased risk of SPMs, while the full impact of an upfront treatment containing both a PI and an IMiD in association with high dose Melphalan (HDM) still has to be investigated. To address this issue, we have evaluated the incidence of SPMs in the GIMEMA 26866138-MMY-3006 multicentre phase III study that compared BOR, thalidomide and dexamethasone (VTD) versus TD as induction before, and consolidation after, a double course of HDM. Methods: Of the 480 transplant eligible NDMM pts enrolled, 474 received assigned treatment. Data on the incidence of SPMs are available for 299 pts (63%, 148 VTD and 151 TD). Results: The median follow up was 73 months. 25/299 pts (8%) developed a SPM: 7 (2%) hematologic and 18 (6%) non hematologic. The median time from trial entry to development of the SPM was 36 months (range 8.4-69.0). The number of pts developing a SPM was lower for VTD (5%) compared to TD (11%, p = 0.068). Among pts developing a SPM, the proportion of solid malignancies was similar between treatment arms (75% and 71% in VTD and TD, respectively). Similarly, hematologic SPMs were 25% for VTD and 29% for TD. Overall the incidence rate (IR) of developing a SPM was 1% at 1 year and 9.9% at 6 years (yrs). This incidence was significantly lower for pts in VTD arm compared with patients in TD (6% vs 13% at 6 yrs, p = 0.037). When looking at the IR of solid tumors only 5% of VTD-treated pts developed a solid SPM, as compared to 9.6% in TD; similarly, less hematologic SPMs were observed in the BOR arm (1% vs 4% at 6 yrs for VTD and TD, respectively). When the analysis was performed according to SPMs type, no statistical significance could be demonstrated. Conclusions: Our data compare favorably with data previously reported on the incidence of developing SPMs in NDMM treated with BOR and Melphalan frontline. With a follow up of 6 years, we were able to confirm that treatment with PI is associated with a low risk of developing SPM. Our data also suggest that treatment with PI might decrease the risk of developing a SPM compared to IMiDs based treatment.

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Does renal failure affect outcome after autologous stem cell transplantation in patients with multiple myeloma? Antlanger M.1, Lamm W.2, Reiter T.2, Porpaczy E.2, Minichsdorfer C.2, Rabitsch W.2, Gisslinger H.2, Agis H.2, Krauth M.-T.2 Medizinische Universität Wien, Innere Medizin III, Abt. für Nephrologie, Wien, Austria, 2Medizinische Universität Wien, Wien, Austria 1

Introduction: Renal impairment (RI) is frequent in patients with Multiple Myeloma (MM) and has been proven to be a prognostic factor regarding overall survival (OS). MM patients with impaired renal function often fail to qualify for high-dose chemotherapy and are excluded from autologous stem cell transplantation (ASCT), since a higher transplant-related mortality has been postulated. However, it remains unclear whether these historical inferior outcome data still hold true in times of modern, immuno-chemotherapeutical therapy regimen. Methods: 195 MM patients (median age 54 years) who had undergone ASCT between 1999 and 2015 were analyzed. Renal function at the time of diagnosis and transplantation was assessed by eGFR (MDRD). Kaplan-Meier curves and log-rank tests were used for OS and progression-free survival (PFS) calculation. Results: Estimated mean overall survival from the time of diagnosis was 93 months (90% CI: 77-109). No difference was found when comparing patients who never had renal impairment with those who presented with renal failure at diagnosis and improved throughout therapy course as well as with those whose renal function was always impaired (Fig 1).

Fig. 1.

Estimated mean PFS was 83 months (CI 12-61). Again, renal impairment did not result in reduced survival (Fig 2).

Disclosure: Annamaria Brioli: Financing of Scientific Research: Celgene, Janssen Michele Cavo: Financing of Scientific Research: Celgene, Janssen

Oncol Res Treat 2016;39(suppl 3):1–104

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tions 5 pts., metabolic disease 5 pts., depression 4 pts., amyloidosis 2 pts., neuropsychiatric disorder 2 pts., status post hematologic malignancies 2 pts., benign hematologic disease 1 pts., disorder of the sensorium 1 pts.. Conclusions: Considering a myeloma incidence rate of 3 - 6 / 100.000 persons and a 5-year prevalence of 17.000 (Germany 2010) outpatient care of 227 patients by 3 hemato-oncological practices within 1 year is strikingly high. Myeloma patients of all stages got initial diagnosis and first as well as following treatment. All approved agents were applied in an outpatient setting (except for high-dose therapy) including multimodal treatments in co-operation with complementary facilities. The schedules applied were conform to latest guidelines and considered latest study results, co-morbidity was carefully regarded. Supportive treatment was fully realized on an outpatient basis. The quality of outpatient care of myeloma patients is to classify as high. Disclosure: Roland Rudolph: Expert Testimony: Fa. Celgene, München Werner Langer: Expert Testimony: Fa. Celgene, München

Posterdiskussion Lymphome P242

Functional identification of novel molecular dependencies in Cyclin D1 driven lymphoma Fig. 2.

Conclusions: In this retrospective analysis, a relatively large cohort of MM patients who had undergone ASCT was analyzed regarding outcome according to renal function. Our data show that neither OS nor PFS after ASCT were negatively impacted by mild to moderate RI. Since exclusion from ASCT results in shorter survival per se, it therefore seems to be of pivotal clinical importance that patients with MM and RI should rather be evaluated pro-actively for high-dose immuno-chemotherapy including ASCT than excluded from these therapy regimen. Disclosure: No conflict of interest disclosed. P241

Outpatient care of multiple myeloma patients in the western Ruhr area. A retrospective assessment from 3 hematooncological practices of related size in Essen, Ratingen and Bottrop Rudolph R.1, Hannig C.2, Langer W.3 Hämato-Onkologische Gemeinschaftspraxis Dres. Rudolph & von Verschuer, Essen, Germany, 2Schwerpunktpraxis für Hämatologie und Internistische Onkologie, Bottrop, Germany, 3Krebszentrum Ratingen, Ratingen, Germany 1

A total of 227 patients were acquired, 86 in Essen, 82 in Bottrop and 61 in Ratingen. 63 pts. were < 65 years old, 73 pts. 65 - 75 years, 91 pts. > 75 years. 114 pts. were female,113 male. Sort of therapy in 2012: watch & wait 174 pts., Bortezomib-based schedules 70 pts., immunomodulatory therapy 53 pts., cytostatic-based therapy 21 pts., high-dose therapy with stem cell transplantation 11 pts., radiotherapy 10 pts., dexamethasone 4 pts.. Supportive treatment in 2012: Bisphosphonates 104 pts., red cell concentrate 36 pts., pain management 36 pts., antibiotics 23 pts., erythropoietin 22 pts., herpes prophylaxis 21 pts., heparin 14 pts., antibiotic prophylaxis 8 pts., radiotherapy 7 pts., ASS 6 pts., platelet concentrate 4 pts., resection of extramedullary lesions 1 pts., denosumab 1 pts.. Co-morbidity in 2012: Arterial vascular disease 140 pts., renal failure 60 pts., diabetes 46 pts., respiratory disorder 22 pts., degenerative skeletal disorder 29 pts., osteoporosis 18 pts., venous vascular disease 17 pts., status post solid tumors 17 pts., peripheral neuropathy 10 pts., solid tumors 10 pts.,osteonecrosis of the jaw 7 pts., dementia 6 pts., endocrinological disorder 6 pts., status post traumatic fractures 5 pts., status post severe infec-

Abstracts

Ehrenfeld S.1,2, Veratti P.1,2, Schneider D.1,2, Duyster J.1,2, Miething C.1,2 Universitätsklinik Freiburg, Department of Medicine I Hematology, Oncology and Stem-Cell Transplantation, Freiburg, Germany, 2DKTK, Standort Freiburg, Freiburg, Germany 1

Mantle cell lymphoma (MCL) presents as a highly disseminated B-cell malignancy with short responses to current therapies and a great need for new therapeutic strategies. The PI3K pathway has emerged as a promising therapeutic target, as MCL cell lines and patients have shown substantial response rates to rapamycin and analogs. We have developed a new mouse model for MCL using Eµ-myc transgene mice that overexpress the MCL hallmark lesion Cyclin D1 together with the reverse tet transactivator. By deriving primary tumor cell lines from diseased mice, we were able to generate a powerful platform for inducible in vitro screening approaches. We performed a functional shRNA-based genetic screen targeting the PI3K pathway to identify novel molecular dependencies in Cyclin D1 driven lymphomagenesis. Using a two colored, antibiotic selectable and tet-inducible retroviral shRNA expression vector system, a library composed of 4 shRNA subpools targeting more than 300 different genes within the PI3K pathway was introduced into murine MCL cells by low-titer retroviral infection. Subsequently shRNA expression was induced by addition of doxycycline and shRNA representation was deconvoluted by NGS to compare variation between knockdown and control cells. Our screen identified a range of proteins that significantly impaired cell proliferation and cell viability. Among these, we found multiple targets within the PI3K pathway and the molecular dependency on this pathway was in line with the observed high sensitivity of these cells towards pharmacological mTOR inhibition. Their mode of action and their impact on the PI3K pathway signaling cascade is currently investigated in more detail. Interestingly, a shRNA targeting the anti-apoptotic protein Bcl-2 showed highly selective depletion in the MCL cells. Since MCL patients have shown particular sensitivity towards a novel Bcl2 inhibitor (ABT199), we are also further exploring the effects of Bcl2 knockdown in Cyclin D1 overexpressing lymphoma. In summary, our shRNA screening approach represents a powerful tool to functionally select novel targets involved in cell proliferation and cell viability, enabling the identification of a range of target genes connected to the PI3K pathway in MCL cells. The newly identified targets will be further investigated in murine and human MCL cells to characterize their role in Cyclin D1-driven lymphomagenesis, as well as their potential as therapeutic targets. Disclosure: No conflict of interest disclosed.

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P243

Clinical and molecular characterization of leukemic follicular lymphomas Viardot A.1, Estenfelder S.1, Tausch E.1, Schrell S.1, Stilgenbauer S.1 Universitätsklinikum Ulm, Klinik für Innere Medizin III, Ulm, Germany

Introduction: Leukemic variants of follicular lymphoma are rare; the clinical significance is unknown. Methods: Chart review of patients with histologically proven follicular lymphoma (FL) and lymphocytosis > 5/nl at diagnosis, treated with chemoimmunotherapy between 2003 and 2015 at a single institution. Results: We identified 23 patients with leukemic FL at initial diagnosis. The median age was 53 years (range 22-74), 70% of patients were younger than 60 years. The initial lymphocyte count was 21.5/nl (range 5.6-154/ nl). The initial histopathological diagnosis was FL Grade 1 (65%), Grade 2 (22%) and Grade 3a or composite lymphoma (13%). In 20/22 patients, immunophenotyping showed CD10 expression. FISH analysis was available in 16/23 patients: the translocation t(14;18)(q32;q21) was detectable in all but 2 patients with tetraploidy on chromosome arm 18q21. The most frequent additional aberrations was gain on 18q21 in 9/16 cases. IGHV status in all analyzed six patients was mutated (homology between 85.5 to 92.7%). The follicular lymphoma international prognostic index was high (median 3), because of stage 4 (100%), elevated LDH (48%), decreased hemoglobin level (43%) and involvement of more than 4 lymph node areas (83%). On the other hand, bulky disease was rare and most patients (16/23) had lymph nodes smaller than 3cm. All patients received chemoimmuntherapy immediately after diagnosis (14 CHOP, 7 Bendamustin, 1 FM and 1 CHOEP, combined with anti-CD20 antibody, respectively). Fifteen patients relapsed after a median time of 16 months. The majority had a nodal relapse; only five patients were leukemic again. The median progression free survival was 14.5 months. Nine patients received autologous stem cell transplantation, and three patients allogeneic stem cell transplantation. With only three deaths (transplant-associated in one patient), the overall survival appeared not dismal. Conclusion: Leukemic FL is a rare variant with typical clinical and molecular characteristics. Patients tend to be younger, have a higher IPI and relapse early after standard chemoimmunotherapy compared to nodal FL. However, overall survival may not be inferior, possibly due to intensive treatment e.g. autologous stem cell transplantation. Disclosure: Andreas Viardot: Advisory Role: Amgen, Janssen, Gilead, BMS; Financing of Scientific Research: Amgen, Roche, Pfizer, Janssen; Other Financial Relationships: Takeda, Janssen Stephan Stilgenbauer: No conflict of interest disclosed. P244

Ibrutinib as first line therapy in a case of Bing-Neel Syndrome Fellas G.1, Kiehl M.1, Hopfer O.1 Klinikum Frankfurt (Oder), Medizinische Klinik I, Frankfurt Oder, Germany

Bing-Neel Syndrome is the rare CNS manifestation of Waldenström macroglobulinemia. In up to 36% of cases it is the first manifestation of the disease and it can present as either a diffuse or tumoral form. Until recently the treatment of this rare entity was based on chemotherapeutics with a good blood-brain penetration, radiation and autologous stem cell transplantation. Ibrutinib, a Bruton’s tyrosine kinase inhibitor, showed efficiency in the treatment of Waldenström macroglobulinemia and good blood-brain penetration. Based on these observations, ibrutinib was used earlier for refractory/relapsed patients with Bing-Neel Syndrome and showed very good results (Cabannes-Hamy A. et al. AJH, 2016). We report here on a case of Bing-Neel syndrome (MYD88L265P) that was treated with ibrutinib 420mg/d. After 4 weeks of therapy a very good response was seen (MRI). No adverse events were noted and the patient’s symptoms have subsided. Ibrutinib has shown a very good efficacy as first line therapy in this case extending its use for the treatment of Bing-Neel Syndrome.

Oncol Res Treat 2016;39(suppl 3):1–104

Fig. 1. MRI imaging before and after ibrutinib therapy. Disclosure: No conflict of interest disclosed. P245

BARs (B-cell receptor antigens for reverse targeting): a novel and ultimately specific treatment concept for B-cell neoplasms Bewarder M.1, Thurner L.1, Fadle N.1, Kemele M.1, Regitz E.1, Neumann F.1, Körbel C.2, Laschke M.2, Preuss K.-D.1, Pfreundschuh M.1 Saarland University Medical School, Dept. Internal Medicine I, José Carreras Center for Immuno- and Gene Therapy, Homburg, Germany, 2Saarland University Medical School, Clinical and Experimental Surgery, Homburg, Germany 1

Introduction: The major task of a B-cell receptor is the binding and internalization of its antigenic target, and its processing for antigen presentation to T-cells. Chronic antigenic stimulation has been discussed to play a role in the pathogenesis of malignant B-cell lymphomas. We therefore systematically searched for the antigenic targets of BCRs from various B-cell neoplasms. Methods: Recombinant BCRs were expressed as recombinant Fabs (rFabs) based on corresponding pairs of genomic VH+VL regions from lymphoma biopsies. Whenever possible, “natural” Fabs (nFabs) were also obtained by papain digestion of fresh or cultured lymphoma cells. Both nFab and rFab were used to screen for binding to recombinant protein macroarrays. Results: Two antigens (hyperphosphorylated paratarg-7 and sumoylated HSP-90 which are modified in patients compared to healthy controls) are targets of paraproteins from (depending on ethnicity) 30-50% of all MM patients; the BCR from 67% of patients with primary CNS lymphoma target hyperglycosylated neurabin, 26% of the BCR from ABC-type DLBCL target hypophosphorylated ARS2 and 45% of all MCL BCR target LRPAP1; optineurin is BCR target of 12% follicular lymphomas and various autoantigens have been identified as targets of roughly 30% of all CLL cases. For all autoantigens binding to its specific BCR, rapid internalization and induction of proliferation was demonstrated. Most importantly, BCR-specific cytotoxicity of recombinant pseudomonas-exotoxin conjugated ARS2 against an ABC-DLBCL cell line with a ARS2 specific BCR was demonstrated in vitro and in vivo after establishment of lymphomas in SCID beige mice. Conclusions: Assuming that only a minority of BCR targets have been identified to date, the prevalence of posttranslationally modified autoantigens strongly supports a role of chronic antigenic stimulation in many B-cell neoplasms. Due to the predominance of a single or few BCR antigens in each malignant B-cell entity studied, BARs represent an attractive and novel therapeutic concept for a broad spectrum of B-cell neoplasms and are the first therapeutic approach in oncology that targets exclusively the malignant cells. BARs can be used for conjugation with toxins, radionuclides and small molecules as well as for bispecific constructs (e. g. with CD3 or CD16) and CAR T-cells, the toxicity of which should be drasti-

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cally reduced due to the ultimate specificity of BARs that spares normal B-cells. Supported by Wilhelm-Sander-Stiftung Disclosure: No conflict of interest disclosed.

P247

Reality of care in Primary Central Nervous System Lymphoma. A retrospective survival analysis of 192 patients at a German University Medical Center Schlosser T.1,2, Illerhaus G.1,3, Schäfer H.S.1 Universitätsklinikum Freiburg, Department Innere Medizin, Klinik für innere Medizin 1, Freiburg, Germany, 2Universitätsklinikum Leipzig AöR, Klinik und Poliklinik für Gastroenterologie und Rheumatologie, Leipzig, Germany, 3 Klinikum Stuttgart, Klinik für Hämatologie, Onkologie und Palliativmedizin, Stuttgart, Germany 1

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Subgroup analyses of diffuse large B-cell lymphoma and indolent lymphoma cohorts from a phase IIa study of singleagent MOR208 in patients with relapsed or refractory nonHodgkin’s lymphoma (R-R NHL) Buske C.1, Jurczak W.2, Zinzani P.L.3, Gaidano G.4, Goy A.5, Provencio M.6, Nagy Z.7, Robak T.8, Maddocks K.9, Ambarkhane S.10, Winderlich M.10, Endell J.10, Blum K.A.9 Institute of Experimental Cancer Research, Comprehensive Cancer Center Ulm, University Hospital Ulm, Ulm, Germany, 2Jagiellonian University, Department of Hematology, Kraków, Poland, 3University of Bologna, Institute of Hematology “L. e A. Seràgnoli”, Bologna, Italy, 4Amedeo Avogadro University of Eastern Piedmont, Division of Hematology, Department of Translational Medicine, Novara, Italy, 5John Theurer Cancer Center, Hackensack University Medical Center, Hackensack, United States, 6University Hospital Puerta De Hierro, Department of Medical Oncology, Madrid, Spain, 7Semmelweis University, First Department of Internal Medicine, Budapest, Hungary, 8Medical University of Lodz, Department of Hematology, Lodz, Poland, 9The Ohio State University Comprehensive Cancer Center, Department of Internal Medicine, Division of Hematology, Columbus, United States, 10MorphoSys AG, Martinsried, Germany 1

Introduction: CD19 is a potential B-cell tumor target. A phase I study showed MOR208, an Fc-engineered, humanized CD19 antibody, to be safe and well-tolerated with encouraging single-agent activity in patients with chronic lymphocytic leukemia. This analysis of our phase IIa study focused on the preliminary efficacy and safety of MOR208 in patients with diffuse large B-cell lymphoma (DLBCL) and indolent (i)NHL. Methods: Patients with R-R NHL progressing after at least one prior rituximab-containing therapy were eligible. Treatment comprised single-agent MOR208, 12 mg/kg IV, weekly, for 8 weeks (2 cycles), which could continue for 4 additional weeks in patients with at least stable disease after 2 cycles. MOR208 maintenance could be given every 2 or 4 weeks until progression in patients with complete (CR) or partial response (PR) at 12 weeks. The primary endpoint was overall response rate (ORR). Results: 92 patients were enrolled including 35 with DLBCL and 45 with iNHL. ORR was 26% in DLBCL (9/35; 2 CRs and 7 PRs), and 27% in iNHL (12/45; 5 CRs and 7 PRs). In evaluable patients (those completing ≥2 cycles, with a response assessment) the ORR was 36% in DLBCL (9/25), and 30% in iNHL (12/40). Median duration of response was 13.7 months for DLBCL (1.2-26; 3 ongoing and >20 months in remission) and 8.4 months for iNHL (2.5-20.3; 6 ongoing, including 1 patient >20.3 months in remission). In DLBCL responders, 5/9 were rituximab-refractory (either no response or response lasting < 6 months to a previous rituximab-containing therapy), of which 2 patients had a response duration of >19.8 months and 1 other of >1 year under MOR208 treatment. The incidence of grade ≥3 hematologic treatment-emergent adverse events was low (DLBCL: 9/35, 26%; iNHL: 4/45, 9%), including neutropenia, anemia and thrombocytopenia in 14%, 9% and 6% of patients with DLBCL and 4%, 0% and 0% of patients with iNHL. Infusion-related reactions were seen in 3/35 (9%) and 4/45 (9%) patients in DLBCL and iNHL cohorts; all were grade 1-2 except for one grade 4 dyspnea. There were no treatment-related deaths and no trend towards late toxicity. Conclusions: Single-agent MOR208 was associated with a promising ORR including CRs and long-lasting responses in DLBCL and iNHL cohorts, including in patients with rituximab-refractory disease. These efficacy data, and the excellent safety and tolerability profile of MOR208 further justify the development of MOR208 as part of a combination therapy for B-cell NHLs. Disclosure: No conflict of interest disclosed.

Abstracts

Introduction: Primary CNS lymphoma (PCNSL) is a highly aggressive non-Hodgkin lymphoma with affection of the neuroaxis. Because of its rarity, studies report about very small numbers of patients often treated within prospective trials. These investigations are often biased by inclusion criteria such as performance status and age. Little is known about the reality of treatment of PCNSL patients in its entirety and only few institutions worldwide provide sufficient data for a broad single-center experience. Methods: A retrospective analysis of 192 consecutive immunocompetent patients with PCNSL seen between 1991 and 2013 at the Department of Hematology and Oncology of Freiburg University Medical Center (Germany) was performed. All ages and every performance status were included to obtain real-life data about the provided healthcare. Kaplan-Meier-estimator was used for survival estimations. Uni- and multivariable analyses were performed to identify predictors for overall survival (OS). Results: The estimated median overall survival (OS) was 31 months (CI 20-65 months). After one, three and five years 64% (CI 58-71%), 48% (CI 41-56%) and 44% (CI 37 52%) of the patients were alive. Age, performance status, LDH serum level, affection of deep brainstructures, Cerebrospinal fluid (CSF) protein level and CSF cell count were significantly associated with OS in univariable testing. Multivariable Cox modelling revealed age, performance status and LDH serum level as independent prognostic factors for OS. The median event-free survival (EFS) time was 10 months (95% CI 7-17 months). Conclusion: Our data confirm known clinical prognostic factors. OS rates are at least comparable if not better than previous reports. In this health service research analysis we report on impressive OS rates in a “real-life” PCNSL patient cohort representing high level of care in these patients at Freiburg University Medical Center. Disclosure: No conflict of interest disclosed. P248

Multicenter case series of primary adrenal lymphoma Majidi F.1, Martino S.1, Haase M.2, Chortis V.3,4, Arlt W.3,4, Spyroglou A.5, Beuschlein F.5, Fassnacht M.6, Kanji A.7, Habra M.A.7, Schott M.2, Haas R.1, Gattermann N.1 Heinrich-Heine University Düsseldorf, Dept. of Hematology, Oncology and Clinical Immunology, Düsseldorf, Germany, 2Heinrich-Heine University Düsseldorf, Dept. of Endocrinology, Düsseldorf, Germany, 3University of Birmingham, Institute of Metabolism and Systems Research, Birmingham, United Kingdom, 4Birmingham Health Partners, Centre for Endocrinology, Diabetes and Metabolism, Birmingham, United Kingdom, 5Ludwig-MaximiliansUniversität München, Medizinische Klinik IV, München, Germany, 6JuliusMaximilian-Universität Würzburg, Medizinische Klinik I, Würzburg, Germany, 7 University of Texas, MD Anderson Cancer Center, Houston, United States 1

Introduction: Primary adrenal lymphoma (PAL) is an extremely rare malignant condition with poor response to therapy and unfavorable prognosis. It is defined as histologically proven lymphoma that involves one or both adrenal glands and presents with no prior history of lymphoma. Methods: We collected 19 patients with PAL from five different centers in Europe and the USA and retrospectively analyzed clinical and pathological features. Results: Median age at diagnosis was 67 (range: 35-82) years. Males were more frequently affected than females (ratio 5:1). The most common symptoms at diagnosis were back pain (29.2%), upper abdominal pain (20.8%)

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and B symptoms (47%, including weight loss in 20.8%). In 2 patients PAL was detected incidentally. Median survival was 8 (1-132) months. Median level of LDH was 542 U/L (210-6515). Bilateral adrenal involvement was detected in 61% and correlated with significantly shorter overall survival: median 3.5 mo. (1.4-11) versus 57 mo. (8-132), with and without bilateral involvement, respectively (P = 0.04). Adrenal insufficiency was diagnosed in 26% of patients, all with bilateral involvement, but had no significant effect on overall survival. In patients with unilateral involvement, the left and right adrenals were affected with similar frequency and resulted in similar clinical picture and prognosis. Mean tumor size (largest diameter) was 9.2 cm (4-16). We found no prognostic impact of tumor size. On histopathology, 17 cases (89%) were diffuse large B cell lymphomas. The remaining 2 cases were a large T cell lymphoma and a NK/T cell lymphoma. Ki67 index was higher than 75% in all 8 cases evaluated. CNS involvement was present in 26%, surprisingly without significant impact on prognosis. In 5 patients another extranodal involvement was detected in lung, kidney (2), duodenum or stomach. Of 13 patients receiving immuno-chemotherapy according to the R-CHOP protocol, 5 obtained complete remission and one achieved partial remission. Conclusion: According to our data, involvement of both adrenal glands is the most important prognostic factor in primary adrenal lymphoma. Up to now, R-CHOP has been the treatment of choice, but the majority of patients fail to respond. Investigations into the molecular pathology of PAL are required to improve our understanding of this rare lymphoid malignancy. It remains to be seen whether modern lymphoma treatments targeting the B cell receptor pathway can improve the outlook of patients with PAL. Disclosure: No conflict of interest disclosed. P249

Decreased ZEB1 expression in Sézary syndrome and its impact on apoptosis and DNA damage response Gand C.1, Grabarczyk P.1, Hirt C.1, Cabron A.-S.1, Przybylski G.2, Iżykowska K.2, Weissmann R.3, Kuß A.W.3, Schmidt C.A.1 Universitätsmedizin Greifswald, Innere Medizin C Hämatologie und Onkologie, Greifswald, Germany, 2Polish Academy of Sciences, Institute of Human Genetics, Poznan, Poland, 3Universitätsmedizin Greifswald Institut für Humangenetik, Interfakultäres Institut für Genetik und funktionelle Genomforschung, ErnstMoritz Arndt Universität Greifswald, Greifswald, Germany 1

Introduction: Sézary syndrome is a rare leukemic form of cutaneous T-cell lymphoma. During the last decades many studies revealed through cytogenetics, comparative genomic hybridization (CGH) and next generation sequencing (NGS) analysis numerous genomic alterations, which are often associated with complex rearrangements. Therefore, a major defect in DNA damage repair mechanisms is probably responsible for the chromosomal instability. The zinc finger, homeodomain transcription factor ZEB1 was recently found to be involved in DNA damage response through Checkpoint kinase 1 (CHK 1). We and others have shown ZEB1 loss as one of the most frequent genetic alterations seen in Sézary syndrome samples. We therefore analyzed the effects of ZEB1 downregulation in T-cell lines focusing particularly on DNA damage response. Methods: To examine the functional consequences of reduced ZEB1 expression, CD4 T-cells from healthy donors and the T-cell lymphoma cell lines Jurkat and HuT 78 were treated with ZEB1 specific siRNAs. Knockdown efficiencies were verified by RT-PCR and Western Blot experiments. Annexin V/ 7AAD double-staining was used for apoptosis detection. Furthermore a ZEB1 deficient Jurkat cell line was established through transduction of ZEB1 specific lentiviral shRNA constructs. DNA damage was induced by Etoposid, Camptothecin and UV-C light. The amount of DNA damage was analyzed through γ-H2A.X staining. Results: An increased spontaneous apoptosis rate was not observed upon siRNA mediated ZEB1 knockdown in healthy T-cells or in the T-cell lines HuT 78 and Jurkat. Furthermore, no influence on chemoresistance upon Camptothecin and Etoposide treatment was found. Interestingly, an increased DNA damage rate could be demonstrated upon treatment with

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UV-C light, Camptothecin and Etoposide in Jurkat cells that were transduced with ZEB1 specific shRNAs compared to scrambled and non-transduced controls. Conclusion: Jurkat cells with reduced ZEB1 expression were prone to DNA-damaging stimulants. Therefore the frequent ZEB1 deletion in Sézary syndrome might be involved in the accumulation of genetic alterations in this disease. These results provide a promising basis for further investigations on the underlying mechanisms of the complex genetic alterations seen in this T-cell lymphoma. Disclosure: No conflict of interest disclosed. P250

A small cell variant of anaplastic large cell ALK positive primary cutaneous T-lymphoma Vergoulidou M.1, Decker T.2, Grobe N.1 Dietrich Bonhoeffer Klinikum, Innere Medizin 1, Neubrandenburg, Germany, Dietrich Bonhoeffer Klinikum, Institut für Pathologie, Neubrandenburg, Germany 1 2

T-lymphomas are rare entities. We report a case of a small cell variant ALK-positive anaplastic large cell lymphoma (ALCL). A 56-year-old male patient presented with multifocal, partially ulcerating skin infiltrates around the right lower and upper eyelid. Biopsy revealed small cell blast morphology. Immunochemistry was negative for CD79a, CD20, CD3, CD4 and CD7 but positive for CD2, cytoplasmatic ALK1 and CD30. CT imaging revealed left axillary lymphadenopathy, so left axillary lymph node dissection was carried out, without histological evidence of lymphoma. Bone marrow biopsy showed no lymphoma infiltration. Patient’s history was remarkable for multiple sclerosis, urothelial carcinoma with excision before ten years and chronic anal fistula. Patient was treated with multi-agent chemotherapy (CHOP) and had complete remission of the skin infiltrates. Primary cutaneous (c-ALCL) and systemic ALK-positive ALCL have identical histopathological findings, further identification is based on clinical findings. Prognosis differs significantly, c-ALCL has 90% longterm survival and only up to 10% extra-cutaneous manifestations. Systemic ALCL small-cell variant has 70% 5-year survival, may present skin infiltrates and presents up to 20% bone marrow infiltration. Our patient fulfilled rather the c-ALCL criteria but due to multifocal skin infiltrates and al least radiologically shown extra-cutaneous manifestation i.e. axillary lymphadenopathy, he received chemotherapy with good tolerability and complete remission. To our knowledge, small cell variant of p-ALCL or systemic ALCL is extremely rare, with only one published case series and sets a diagnostic as well as therapeutical challenge. Disclosure: No conflict of interest disclosed. P251

ALK-negativ anaplastic large cell-lymphoma – a supprising diagnosis in a patient with low grade prostatic cancer: a case report Piribauer M.1, Weiss H.1, Tomka M.1, Siebert F.1 Krankenhaus der Barmherzige Brüder, Innere Medizin, Hämato-Onkologie, St. Veit/Glan, Austria 1

Introduction: Prostatic cancer is the most frequent tumor in males. Anaplastic large cell-lymphoma (ALCL) are a rare hematological malignancy and a distinct subtype of mature T-Cell lymphomas. We presenting a 72year-old patient with liver and lung lesions where the histological investigation shows an ALK- negative ALCL four years after diagnosis of a low grade prostatic cancer. Case report: A 72year-old male patient was admitted to our hospital due to newly diagnosed lesions in the liver and lung. Four years before he had radical prostatectomy and lymph node removal because of prostatic cancer. Histologic Adenocarcinoma, Gleason Score 3+4/7, pT2c, N0, R0. The

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histological investigation of tissue probe from the bronchoscopy could not fount malignant cells. Only cytological investigation shows cells of an adenocarcinoma, G2. So the next procedure was a puncture of one liver leasion. Histological shows now a solid growing cancer, the tests of immunohistochemistry were outstanding. In the meantime the patient general condition and dyspnea worsened. With the suspicion of a lung cancer (NSE with 32, 6 µg/l elevated (normal range: < 18,3 µg/ml) PSA was in normal range), we started chemotherapy with cisplatin and vinorelbine. After one cycle the patient shows a clinical benefit, dyspnea improved. Days later, the result of immunohistochemistry shows no polymorphic carcinoma and no markers for prostatic cancer. Negative were ALK 1and TIA 1 and CD 20. Positive was CD 30 and the T-cell markers CD3 and CD5. Now the diagnosis of an ALK-negative Anaplastic T-cell-lymphoma, IPI-score high risk, was found. Chemotherapy was switched to COMP-scheme. The patient tolerated chemotherapy well, the dyspnea improved rapidly. Now, four cycles was administrated (planned six cycles), the patient are in well condition. Conclusions: To our knowledge, is this the first case of an ALK- negative ALCL in a male patient with low grade prostatic cancer. After literature search, ALCLs were often described in female patients with breast implantants, some in pancreatic, lung and skin and rarely in leukemia. Among all systemic ALCLs, those that are ALK negative constitute 1550% of cases. The prognosis is poorer than in ALK-positive ALCLs. The recommended chemotherapy is anthracycline based. It usually involves lymph nodes at diagnosis (49%) and, less frequently, extranodal sites (20%). This case also underlines the importance to puncture lesions for histological investigation in unclear cases. Disclosure: No conflict of interest disclosed. P252

Allogeneic stem cell transplantation in refractory Hodgkin’s lymphoma after CR with Nivolumab Fellas G.1, Kiehl M.1, Hopfer O.1 Klinikum Frankfurt (Oder), Frankfurt Oder, Germany

Hodgkin’s lymphoma is a hematologic malignancy, characterized by the presence of the Reed-Sternberg cells within an inflammatory infiltrate. Although it is a highly curable disease, approximately 10-15% of cases are primary refractory to the convenient therapies with subsequently poor outcomes. Recurrent chromosome 9p24.1 amplification in Reed-Sternberg cells results in overexpression of the PD-1 ligands (PDL1 and PDL2) with subsequent T cell exhaustion. Recently, Nivolumab, a novel fully human monoclonal IgG4 antibody directed against PD-1, showed very good remission rates in patients with refractory disease. We report the cases of two previously heavily treated young patients with refractory Hodgkin’s lymphoma. Both patients were treated with Nivolumab 3mg/kg every 2 weeks. One patient showed primarily a good response but he was progressive after 9 cycles and the therapy was discontinued. However, the other patient reached a complete response (CR) after 8 cycles of Nivolumab therapy and was able to proceed to consolidating allogeneic stem cell transplantation. After 200 days the patient is still in CR without any significant complications. The response to treatment was very different raising the issue of response prediction. In the post-transplant period there was seen no increased acute GvHD risk, stressing the concept of PD-1 blockade before and possibly also after allogeneic transplantion in r/r Hodgkins lymphoma. We could show here that PD-1 check point blockade can lead to a complete remission (CR) in previously refractory Hodgkin’s lymphoma and that subsequent consolidating allogeneic stem cell transplantation is possible with acceptable adverse events.

Fig. 1. CR after Nivolumab.

Disclosure: No conflict of interest disclosed. P253

A comprehensive analysis of seasonality patterns of incidence and mortality in Hodgkin lymphoma across geographic regions Borchmann S.1, Müller H.1, Engert A.1 Uniklinik Köln, Klinik I für Innere Medizin, Köln, Germany

Introduction: The goal of this analysis is to provide a comprehensive analysis of seasonal variation in incidence and mortality risk of HL with a focus on latitudinal differences. Methods: HL cases diagnosed between 1973 and 2012 in the 18 Surveillance, Epidemiology, and End Results (SEER) registries were eligible (n = 50,179). After exclusion of cases with partly missing data (n = 8,700) and the SEER registries “Alaska natives” (n = 17) and “Rural Georgia” (n = 57) due to insufficient case numbers, 41,405 cases were included. Seasonality of incidence was analyzed for all cases and predefined subgroups using cosinor analysis. The overall mortality risk following a HL diagnosis in winter (Sep.-Feb.) vs. summer (Mar.-Aug.), after correction for known risk factors, was compared using a Cox proportional-hazards model. The interaction between seasonal mortality differences and latitude was analyzed. Results: HL shows a seasonal incidence pattern with a peak around March and a trough around September (p < 0.001). Only cases of the mixed cellularity (p < 0.001), nodular sclerosis (p < 0.001) and lymphocyte depleted subtype (p = 0.002) showed a seasonal incidence pattern. The estimated amplitude of seasonal differences between the peak and the trough of monthly incidence was particularly high for the age groups 20-29, 30-39 and 60-69, with amplitudes of 0.121, 0.114 and 0.096 respectively, coinciding with age groups of increased HL incidence. Cases diagnosed at lower (< 38.05°N) latitudes showed a decreased seasonality of incidence with an amplitude (0.055) compared to cases at higher (≥38.05°N) latitudes (0.102) (p = 0.023 for difference). The risk of dying within the first 3 years after a HL diagnosis in winter is increased at higher (≥38.05°N) latitudes (HR: 1.082 [1.009;1.161], p = 0.027), whereas no seasonal difference in mortality was observed for cases diagnosed at lower (< 38.05°N) latitudes (HR: 0.990 [0.926;1.059], p = 0.772). Conclusion: Increased seasonality of incidence at higher latitudes and the interaction between higher latitudes and increased mortality after a HL diagnosis in winter show the influence of latitude on these patterns. As latitude is closely linked to seasonal variations in Vitamin D serum levels, a protective effect of Vitamin D in HL is a possible explanation. Further evidence, especially on the direct association between Vitamin D levels and the clinical course of HL, needs to be collected to improve the understanding of the role of Vitamin D in HL. Disclosure: No conflict of interest disclosed.

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Metamorphosis of a lymphoma – from extranodal marginal zone lymphoma over a diffuse large B-cell lymphoma to Hodgkin lymphoma – a case presentation Potenberg J.1, Reyher-Klein S.2 Onkologisches Zentrum Ev. Waldkrankenhaus, Berlin, Germany, Pathologisches Institut am Ev. Waldkrankenhaus, Berlin, Germany

1 2

Introduction: The prognosis of localized gastric malt lymphoma is apparently good. We present here a case of lymphoma whose dangerousness has been judged differently during treatment and finally took a fatal outcome. Methods: To provide a high quality of our diagnostic and treatment we try to include our patients in studies and obtain a second opinion in difficult cases. Results: The previously healthy patient (born 1961) was diagnosed in January 2011 with a gastric malt lymphoma, stage E II. The patient was included in a study performed by the German Gastrointestinal Study Group. After eradication of H. pylori, radiotherapy including tomotherapy of the stomach was performed and remission was achieved. May 2013 the lymphoma inthe stomach recurred. The biopsy showed now areas of diffuse large cell b-cell lymphoma. 6 cycles of R-CHOP 14 were given and a complete remission was obtained. November 2015 the patient developed multiple lymphomas. An axillary node was exstirpated and the diagnosis was Hodgkin lymphoma with nodular sclerosis. Chemotherapy with BEACOPP escalated was applied. During therapy a further progress was noted. The therapy was switched to DHAP. During the second cycle of DHAP there was again progression. Brentuximab-vedotin was given. After the first cycle the patient got pneumonia and died despite the treatment in an intensive care unit. Conclusions: A case is presented here that showed a development from extranodal marginal zone lymphoma over a diffuse large B-cell lymphoma to Hodgkin lymphoma. Despite the wealth of opportunities our patient died of the lymphoma.

Fig. 1. Numerous small polyps seen during colonoscopy.

Disclosure: No conflict of interest disclosed. P255

Unusual presentation of Hodgkin’s lymphoma Dickmann J.R.M.1, Kerstan H.2, Heine M.3, Morche M.4, Austein T.1 St. Bernhard Hospital, Department of Haematology and Oncology, Brake, Germany, 2St. Bernhard Hospital, Department of Gastroenterology, Brake, Germany, 3Institute of Pathology, Bremerhaven, Germany, 4Institute of Radiology Jade Weser, Brake, Germany 1

Introduction: Lymphomas typically present with peripheral lymph node swelling. Occasionally they may manifest within other lymph node regions or organs1. We report an unusual case of Hodgkin’s lymphoma affecting the skin and colon. Case: A 42-year-old man presented to our department with general decline, night sweats and dyspnoea. Examination found generalised lymphadenopathy and a right pectoral skin exanthema. Laboratory data showed a normocytic anaemia (Hb 7,7 g/dl, MCV 78fl) with a leuko- and thrombocytosis. A colonoscopy found numerous polyps in the sigmoid colon. Histologically these contained atypical CD30+ cells. Skin, lymph node and bone marrow biopsies were all consistent with a diagnosis of Hodgkin’s lymphoma. Radiological investigations suggested the involvement of the lung parenchyma. Due to the patient’s poor physical condition, we started chemotherapy with ABVD. After improvement of the patient’s condition, we escalated the therapy and he achieved a complete remission after four cycles of BEACOPP.

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Fig. 2. Reed-Sternberg cell from sigmoid polyp biopsy.

Discussion: Hodgkin’s lymphoma is rarely found to manifest in the gastrointestinal tract. Unfortunately the impact on prognosis is not known. There are a few cases described in the literature of primary gastrointestinal Hodgkin’s lymphoma2 presenting with a stricture or ulceration. Our patient gave no history of gastrointestinal symptoms and our findings were unexpected. More data is needed to ascertain what impact this manifestation has on the prognosis for patients. Conclusion: When patients with a suspected diagnosis of lymphoma present with a severe anaemia, endoscopic investigations may be considered to rule out gastrointestinal involvement. References: 1 Austein T, Demme B: Uncommon recurrence of follicular lymphoma, Eur J Haematol. 2012 Jan;88:91. 2 Sharma S. et al. Primary Intestinal Hodgkin´s Lymphoma: An Uncommon Presentation. J Lab Physicians. 2013 Jul;5:124–126. Disclosure: No conflict of interest disclosed.

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HIV-patient with advanced Hodgkin Lymphoma, hyperinflammation and limited NSCLC Mueller M.1, Rittweger M.1, Weber C.1, Lindner A.1, Arasteh K.1 Klinik für Infektiologie und Gastroenterologie, Vivantes Auguste-ViktoriaKlinikum, Berlin, Germany 1

Introduction: We report a case of a HIV-positive patient with classical Hodgkin-Lymphoma (HL), severe hyperinflammation and a limited NSCLC. Method: Case report. Results: A 66y male patient with recently diagnosed HIV-infection (CD 95/µl, 33%, viral load 8600 c/ml), a history of syphilis and tobacco use of 60 packyears, was admitted in our hospital because of fever, encephalopathy, pancytopenia and hypoalbuminemia. The seriously ill patient was transferred to the ICU with hyperinflammation. 6 of 8 criteria for hemophagocytic lymphohistiocytosis (HLH) were positive (Henter 2007). A classical EBV-associated Hodgkin Lymphoma was diagnosed with bone marrow, liver and lymph node involvement. CT-scan and biopsy revealed also a peripheral lung tumour (large-cell neuroendocrine carcinoma). Patient recovered quickly after one cycle of ABVD (dose-adjusted because of hyperbilirubinemia). A complete remission of the HL was achieved after 5 cycles of BEACOPP-baseline and PET-CT showed also a partial remission of the NSCLC. A HSV2 encephalitis were treated successfully and the NSCLC was resected subsequently (pT2b,N1,V1,R0,G3). After tumour board discussion 4 cycles of Cisplatin/Etoposid (dose reduction because of thrombocytopenia and febrile neutropenia) were applied. He died unexpectedly shortly after the last cycle. Autopsy revealed heart failure. Conclusion: -In HIV-positive patients with fever of unknown origin HLH due to Hodgkin-Lymphoma should be ruled out. -Second neoplasia in an immunosuppressed patient is not a contraindication for curative treatment. Disclosure: No conflict of interest disclosed.

Posterdiskussion

Allogene Stammzelltransplantation 1

er on CD8+ T cells to determine the human regulatory CD8+ T cell subset. Similar to murine CD8+ Treg, the cells express CD44, CD122 as well as the transcription factor Helios. In vitro assays showed responsiveness of KIR+CD8+ cells to IL-15 and suppression of CD4+ TFH cell responses. Assessment of KIR+CD8+ T cells in patients after allo-SCT revealed the presence of CD8+ Treg as early as day +30 after transplantation. The proportion of the CD8+ Treg subset increases thereafter and reaches a peak around day +90. This T cell population has a stable expression of the surface markers CD44 and CD122 whereas the transcription factor Helios is significantly up-regulated after allo-SCT when compared to healthy donors. Taken together, we have identified that Helios+KIR+CD8+ Treg population recover early after allo-SCT. Our findings implicate an important role of CD8+ Treg during regeneration of the immune system in these patients and may control development of GvHD. Systematic quantification of KIR+CD8+ T cells in patients after allo-SCT may provide a deeper insight into the functional involvement of this Treg population in the pathogenesis of acute and chronic GvHD. These data will then set the stage for potential immune-modulatory strategies specifically activating or expanding CD8+ Treg to prevent and/or treat GvHD. Disclosure: No conflict of interest disclosed. P258

Trephine biopsies are preferable to isolate mesenchymal stromal cells after allogeneic hematopoietic stem cell transplantation Krüger T.1, Middeke J.M.1, Stölzel F.1, Mütherig A.1, List C.1, Brandt K.1, Heidrich K.1, Döpper J.1, Teipel R.1, Francke S.1, Müller K.1, Kräter M.1, Dhawan A.1, Ordemann R.1, Schuler U.1, Oelschlägel U.1, Wermke M.1,2, Wehner R.3, Schmitz M.3,4,5, Bornhäuser M.1,4,5, von Bonin M.1,5,6 Medizinische Klinik und Poliklinik I, Universitätsklinikum Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany, 2Universitäts Krebs Centrum (UCC), Early Clinical Trial Unit (ECTU), Universitätsklinikum Carl Gustav Carus, Dresden, Germany, 3Institut für Immunologie, Medizinische Fakultät, Technische Universität Dresden, Dresden, Germany, 4Nationales Centrum für Tumorerkrankungen (NCT), Partnerstandort Dresden, Dresden, Germany, 5 Deutsches Konsortium für Translationale Krebsforschung (DKTK), Standort Dresden, Dresden, Germany, 6Deutsches Krebsforschungszentrum (DKFZ), Heidelberg, Germany 1

Graft-versus-Host Disease (GvHD) is a potentially life-threatening complication after allogeneic stem-cell transplantation (allo-SCT), limiting the therapeutic success of this paradigmatic immunotherapeutic approach to treat hematological diseases. CD4+ regulatory T cells (Treg) have been shown to be beneficial in the prevention and/or treatment of GvHD while sparing the Graft-versus-Leukemia (GvL) effect. The contribution of regulatory CD8+ T cell-mediated immune responses in GvHD, however, has not been studied so far. Recent studies have identified a small subset of IL15 dependent CD8+ Treg that are essential for maintenance of self-tolerance and which is characterized by the co-expression of CD44, CD122 and Ly49. The differentiation and the immune-regulatory function of CD8+ Treg depend on the transcription factor Helios. Here we have identified the early development of CD8+ Treg after allogeneic stem-cell transplantation. We defined the human homologue of murine CD8+ Treg using the killer cell immunoglobulin-like receptor (KIR), the functional homologue of the murine Ly49 receptor, as a surface mark-

Introduction: Mesenchymal stromal cells (MSCs) substantially participate in maintaining hematopoiesis as self-renewing progenitors of several different stromal cells of the bone marrow microenvironment and as part of the hematopoietic stem cell (HSC) niche. Because bone marrow stromal cells remain host-derived after allogeneic stem cell transplantation (SCT) it is suggested that the HSC niche could be a target of Graft versus Host Disease (GvHD). Indeed myelosuppression has been repeatedly described in the context of GvHD and in particular a low platelet count was proposed as an indicator of GvHD severity. Hence MSCs as part of the HSC niche may play a key role in development and modulation of GvHD associated myelosuppression. Therefore, it would be clinically relevant to investigate these cells in this context. However, bone marrow aspirates usually used to isolate MSCs fail to reliably recover sufficient amounts of cells after allogeneic SCT. Methods: Bone marrow aspirates (5-10ml) and trephine biopsies were performed within routine diagnostics after allogeneic SCT. Both, aspirates and trephine biopsies were processed using a collagenase based protocol. The colony-forming unit – fibroblast (CFU-F) assay was used to functionally detect MSCs. MSC frequency was determined by limiting dilution assay. Remaining cells were seeded for expansion. Results: Limiting dilution assay of 21 pairs of aspirates and trephine biopsies revealed a significant higher MSC frequency in trephine biopsies (trephine biopsy: 1 MSC in 1.5×104 ± 2.7×104 TNCs; aspirates: 1 MSC in 8.97×105 ± 8.27×105 TNCs, p = 0.0002). Furthermore, expansion of MSCs isolated from trephine biopsies was more reliable and led to a significantly higher yield at passage 0 compared to aspirate derived MSCs.

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Reconstitution of CD8+ Treg after allogeneic stem-cell transplantation Holderried T.A.W.1, Sauerborn P.1, Kim H.-J.2, Cantor H.2, Wolf D.1, Brossart P.1 University Hospital Bonn, Department of Hematology, Oncology and Rheumatology, Bonn, Germany, 2Harvard Medical School, Department of Cancer Immunology and Virology, Boston, United States 1

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Conclusion: Collagenase digestion of trephine biopsies is a suitable method to reliably isolate sufficient numbers of MSCs after allogeneic SCT which now allows the examination of their role in GvHD-mediated myelosuppression. Disclosure: No conflict of interest disclosed. P259

Chip-based digital PCR: an accurate and sensitive method for routine chimerism monitoring after hematopoietic stem cell transplantation Gourri E.1, Schanz U.2, Frey B.M.1, Gassner C.1 Blood Transfusion Service Zurich, Swiss Red Cross, Department of Molecular Diagnostics and Research & Development (MOC), Zurich-Schlieren, Switzerland, 2 University Hospital Zurich, Department of Hematology, Zurich, Switzerland 1

Introduction: After hematopoietic stem cell transplantation (HSCT) recipient and donor populations of leukocytes cohabit in the peripheral blood of the patient, resulting in “chimerism”. Monitoring chimerism is crucial to follow the outcome of the disease and to make informed clinical decision concerning further therapeutic interventions. Current chimerism monitoring is performed using Short-Tandem-Repeats analysis (STR) and quantitative real-time PCR (RT-PCR). However, both methods suffer from limited sensitivity and reproducibility. Digital PCR (dPCR) represents a new alternative for accurate and reproducible chimerism monitoring. Methods: Chip-based Quantstudio 3D chip-based dPCR (Applied Biosystems, Reinach, Switzerland) allows for SNP detection in approximately 20’000 separate RT-PCRs per sample, with statistical presence of only 0, 1, or 2 DNA molecules per each 865 pL reaction well. Fluorescence of each well is separately measured and delivers absolute counts of the two different alleles. Original samples were analyzed for their SNP genotypes by PCR using Sequence Specific Priming (PCR-SSP). Results: Using artificial DNA mixes, we estimated our limit of detection to range at approximately 0.5% for the minor DNA. Results within the same samples were highly homogeneous using six different SNP-Taqman assays for evaluation. Typing samples of Instand’s external proficiency testing (EPT) for post-HSCT chimerism, was performed on one preHSCT patient and donor sample each and on five post-HSCT samples, using only two different SNP-Taqman assays. Instand honored our result quality with 20/20 points. On 15 DNA sample triplets, consisting of one pre-HSCT, one donor and one post-HSCT specimen each, dPCR results were consistent with previous STR results in 11 out 15 triplets. Discrepancies were only observed for 4 triplets, where chimerism was estimated to range below 5% by STR analysis, a method with known limited sensitivity (Stahl et al., Exp Hematol, 2015 Jun). Conclusion: According to the high performance of our method in the Instand EPT and based on the results of 15 patient/donor/post-HSCT sample triplets, chip-based dPCR appears as an accurate and routinely applicable technique for exact chimerism determination with excellent sensitivity. Disclosure: No conflict of interest disclosed. P260

IL17 as a marker for acute GVHD Wolff D.1, Schmitt T.1, Wölfinger P.1, Bode S.1, Theobald M.1, Wagner E.M.1 UCT Universitätsmedizin Mainz, Hämatologie, internistische Onkologie, Pneumologie, Mainz, Germany 1

Introduction: Allogeneic hematopoietic stem cell transplantation (HSCT) is a curative option for patients suffering from AML. Despite of the beneficial graft versus leukemia effect the Graft versus host disease (GVHD) as a dysregulation of the donor immune cells is one of the major complications after HSCT and causes morbidity and mortality. Major players in this immune reaction are donor T cells (CD8 and CD4 T cells), but the mechanism is not fully understood. Especially the role of CD4 T

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cells remains unclear. Here we investigate on IL17-producing CD4 T- cells in the course of GVHD in our patients after allogeneic HSCT. IL17 producing cells are controversial in literature, their impact in development of GVHD is under discussion. Methods: We analyzed PBMC´s from patients after allogeneic HSCT and dose reduced conditioning regimens with multicolor FACS-staining for CD3, CD4, CD8, and regulatory T-cells (CD25, CD127neg, FoxP3) as well as intracellular IL17. IL17 production was measured with and without stimulation with PMA. Our patient cohort consists of 21 patients with acute GVHD, 13 patients with chronic muco-cutaneous GVHD and 5 healthy controls. For 3 patients with overlap syndrome we collected blood samples in the time course of GVHD and correlated our findings with clinical outcome and response to treatment. Results: At onset of GVHD we found no significant differences in the absolute and relative amounts of CD3, CD4, CD8 and Treg counts in our patients with acute or chronic GVHD. Also the unstimulated baseline secretion of IL17 was similar in patients with acute or chronic GVHD compared to healthy controls. In contrast, after stimulation with PMA the increase in IL17 positive T cells was significantly higher in patients with acute GVHD compared with patients with chronic GVHD or healthy controls. Especially in patients with severe gastrointestinal GVHD we found highest percentages of IL17-producing CD4-T cells. In patients with overlap syndrome, these amounts of IL17 positive CD4-Tcells correlates with treatment refractory persisting GVHD. Conclusion: In patients after allogeneic stem cell transplantation the increase in IL17 producing CD4 T-cells after PMA-stimulation correlates with acute GVHD and treatment response. This biomarker should be investigated in a prospective manner to diagnose GVHD and could guide immunosuppressive treatment. Disclosure: No conflict of interest disclosed. P261

Results of sequential therapy in the setting of unmanipulated HLA-haploidentical transplantation utilizing posttransplantation high-dose cyclophosphamide in the treatment of high-risk AML and MDS Prevalsek D.1, Fritsch S.1, Hubmann M.1, Zoellner A.-K.1, Köhnke T.1, Engel N.1, Bücklein V.L.1, Schulz C.1, Mumm F.1, Berking S.1, Ledderose G.1, Stemmler H.-J.1, Spiekermann K.1, Hiddemann W.1, Hausmann A.1,2, Tischer J.1 Klinikum der Universität München – Campus Grosshadern, Medizinische Klinik und Poliklinik III, München, Germany, 2Klinikum München-Schwabing, Medizinische Klinik I, München, Germany 1

Hematopoietic stem cell transplantation offers a curative treatment option for patients suffering from high-risk AML and MDS, but the availability of HLA-matched donors might be limited. To evaluate the feasibility and outcome of sequential therapy in the context of unmanipulated HLA-haploidentical hematopoietic stem cell transplantation (haplo-HSCT) utilizing post-transplantation cyclophosphamide (PTCY) in patients with high-risk, relapsed and refractory AML/ MDS, we retrospectively evaluated the course of 64 patients (AML n = 61, MDS n = 3; median age: 50 years) transplanted between 2009 and 2015 at our center. Disease was advanced in 55 patients including 24 patients with relapse after a first allogeneic transplantation. 54 patients presented with active disease at time of haplo-HSCT while 9 patients were in CR. All patients received sequential therapy combining cytoreductive chemotherapy (clofarabine n = 34; FLAMSA n = 25; FLAG-Ida n = 2; others n = 3) and reduced-intensity conditioning (RIC) which was started after three days of rest thereafter. Conditioning was drug-based in the majority of patients (n = 42). As backbone of post-grafting immunosuppression high-dose cyclophosphamide was given on day + 3 and +4 in all patients. Bone marrow was the graft source in 37 patients. One graft rejection was observed. Neutrophil/platelet engraftment was achieved in 58/49 patients, respectively. Acute GvHD grade II-IV occurred in 19 patients (30%) while it was severe (grade III-IV) in only 3

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(5%). Chronic GvHD was most frequently mild (n = 9) to moderate (n = 8); one severe chronic GvHD occurred. Severe (grade III-IV) mucositis, hemorrhagic cystitis and hand-foot syndrome/rash were observed in 10, 5 and 2 patients, respectively; no patient developed VOD. Kidney failure requiring hemodialysis occurred in 7 patients. CMV reactivation was observed in 59%, while only one patient developed CMV disease (pneumonia) and no patient developed PTLD. Probable or proven (n = 2) invasive aspergillosis was diagnosed in 10 patients. One-year non-relapse mortality was 27.5%. After a median follow up of 21 months (range: 3-64), estimated two-year OS and DFS were 39%, both. In summary, sequential therapy in the setting of TCR haplo-HSCT using PTCY is well tolerated with low rates of GvHD and acceptable NRM in high-risk AML/MDS, while providing an effective anti-leukemic activity in advanced disease. Thus, we suggest that donor availability can be expanded in patients with high-risk AML/MDS. Disclosure: No conflict of interest disclosed. P262

Graft-versus-leukemia effects in T-prolymphocytic leukemia: evidence from minimal residual disease kinetics and T cell receptor repertoire analyses Sellner L.1, Brüggemann M.2, Schlitt M.2, Knecht H.2, Herrmann D.2, Reigl T.3, Krejci A.3, Bystry V.3, Darzentas N.3, Rieger M.4, Dietrich S.1, Zenz T.1, Ho A.D.1, Kneba M.2, Dreger P.1 Universitätsklinikum Heidelberg, Medizinische Klinik V, Heidelberg, Germany, 2Universitätsklinikum Schleswig-Holstein, Kiel, Germany, 3Central European Institute of Technology, Brno, Czech Republic, 4Onkologische Schwerpunktpraxis Darmstadt, Darmstadt, Germany 1

Introduction: Allogeneic stem cell transplantation (alloSCT) may provide long-term disease control in T-prolymphocytic leukemia (T-PLL). However, direct evidence of graft-versus-leukemia (GVL) activity in T-PLL is lacking. We investigated GVL by correlating minimal residual disease (MRD) kinetics with immune modulatory interventions and T cell receptor (TCR) repertoire diversity after alloSCT. Methods: The study sample consisted of 10 patients who received alloSCT for T-PLL at the University of Heidelberg between 2007 and 2015. Quantitative MRD monitoring was performed using clone-specific real-time quantitative PCR (RQ-PCR) of clonal TCR beta (TRB) and/or gamma gene rearrangements. TCR repertoire diversity was analyzed longitudinally by next-generation sequencing (NGS) on Illumina’s MiSeq platform. Results: The 3-year relapse-free and overall survival was 48% (95%CI 1680%) and 58% (95%CI 27-90%), respectively. All patients had a cytological complete response (CR) after alloSCT (5 unrelated, 4 related, 1 haploidentical). 2 patients died early because of acute GvHD, and one had no MRD marker, leaving 7 patients for MRD monitoring. Of these, 3 were MRD- at alloSCT, whereas 5 patients remained or became MRD+ early after alloSCT. In all of these 5 patients, immunosuppression tapering (3) or DLI (2) resulted in significant reduction of MRD levels (range 1-3 log) and was accompanied by cGvHD in 3 patients. However, durable MRD- was obtained in only 2 patients (alive 86+ and 12+ months post transplant). The TRB repertoire of the three patients with the longest follow-up (up to six years) was interrogated longitudinally using NGS. In all patients, MRD decline was reproducibly associated with a shift from a clonal, T-PLL-driven profile to a polyclonal signature which largely corresponded to the donor TCR repertoire and receded with increasing MRD levels. Notably, there was no obvious correlation of GVL-induced MRD decline with emergence of dominant T cell clonotypes that could explain a clonal GVL effect. Conclusions: This study provides first direct evidence for GVL activity in T-PLL, even though it appears to be often only limited or transient. Moreover, GVL in T-PLL does not seem to be driven by the emergence of novel dominant T cell clones but is rather relying on poly- or oligoclonal T cell responses. Nonetheless, alloSCT in T-PLL is a valuable treatment option, and further evaluation of MRD monitoring is warranted to optimize patient care after alloSCT.

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Proteasome inhibitors and immunomodulatory drugs improve survival in relapsed or progressive multiple myeloma after allogeneic hematopoietic cell transplantation Schneidawind C.1, Duerr-Stoerzer S.1, Faul C.1, Kanz L.1, Weisel K.1, Bethge W.1, Schneidawind D.1 Department of Medicine II, Eberhard Karls University, Tübingen, Germany

Introduction: Multiple myeloma is the most common indication for autologous hematopoietic cell transplantation (HCT) but transplantation from an allogeneic donor is controversial and reserved for high-risk patients with refractory, relapsed or progressive disease. Methods: We report a retrospective single center analysis of 41 consecutive patients that underwent allogeneic HCT for the treatment of multiple myeloma from 2001 to 2015. Results: We identified and analyzed 41 consecutive patients with a median age of 53 years (range, 35-67) transplanted for the treatment of relapsed (n = 11), progressive (n = 27) or refractory (n = 3) multiple myeloma. Median time from diagnosis of multiple myeloma to allogeneic HCT was 34 months (range, 2-142). Median time from first autologous HCT to allogeneic HCT was 21 months (range, 1-107). The number of patients transplanted from matched unrelated donors (MUD, n = 20) was high compared with matched related donors (MRD, n = 13) or mismatched unrelated donors (MMUD, n = 8). 3-year event-free survival (EFS) and overall survival (OS) of the whole cohort were 15% and 51%, respectively. Allogeneic HCT after a second high-dose chemotherapy with autologous stem cell support (n = 18) was associated with a decreased 3-year EFS (6% vs. 24%, p = 0.04) and OS (35% vs. 64%, p = 0.09). Cumulative incidence of non-relapse mortality (NRM) at 3 years was 20%. However, the 3-year cumulative incidence of relapse or progression adjusted for NRM as competing risk was 65%. In case of relapse or progression after allogeneic HCT, the treatment with proteasome inhibitors or immunomodulatory drugs significantly improved survival (1-year OS 79% vs. 29%, p = 0.001). Importantly, the administration of immunomodulatory drugs and/or proteasome inhibitors was well tolerated. We did not observe acute graftversus-host disease (GVHD) ≥2 but limited and extensive chronic GVHD occurred in 5 (25%) and 3 (15%) patients that received a lenalidomid-containing regimen. Conclusions: Our data suggest that proteasome inhibitors and immunomodulatory drugs are important for the management of relapsed and progressive multiple myeloma following allogeneic HCT. The incorporation of these agents into transplant protocols might improve outcomes and refine the role of allogeneic HCT for the treatment of multiple myeloma as a platform for long-term disease control. Disclosure: No conflict of interest disclosed. P264

Central nervous system complications after allogeneic hematopoietic cell transplantation – a significant cause of morbidity and mortality Simon C.1, Zierhut M.2, Mirza N.1,3, Faul C.1, Korn A.4, Vogel W.1, Rammensee H.-G.3, Kanz L.1, Bethge W.A.1, Haen S.P.1,3 Medizinische Universitätsklinik Tübingen, Abteilung II für Onkologie, Hämatologie, Immunologie, Rheumatologie und Pulmologie, Tübingen, Germany, 2Universitätsaugenklinik Tübingen, Tübingen, Germany, 3 Interfakultäres Institut für Zellbiologie, Abteilung Immunologie, Tübingen, Germany, 4Universitätsklinik für Neuroradiologie, Tübingen, Germany 1

Disclosure: No conflict of interest disclosed.

Introduction: Central nervous system (CNS) complications after allogeneic hematopoietic cell transplantation (HCT) can represent diagnostic and therapeutic challenges since differential diagnosis between malignant, inflammatory, circulatory or infectious causes is difficult and time-consuming but immediate precise therapy is mandatory to prevent permanent neurological damage. Methods: Out of 1,204 patients undegroing allogeneic HCT at our institution between 2003 and 2015, we identified 102 patients (8.6%) with

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post-transplantation CNS complications. The patients (38 women and 64 men, median age 55 years, range 19-75 years) received HCT due to acute myeloid (n = 49) or lymphoblastic (n = 24) leukemia, chronic lymphocytic (n = 2), myelomonocytic (n = 2) or myeloid (n = 1) leukemia, lymphoma (n = 10), myelodysplastic (n = 8) or myeloproliferative (n = 5) syndromes, as well as severe aplastic anemia (n = 1). Conditioning was performed using reduced intensity (n = 61) or myeloablative (n = 41) regimens. Results: CNS complications comprised bleeding (n = 23) or thromboembolic (n = 11) events, infections (n = 19), toxic leukoencephalopathy (n = 10), inflammatory changes without detection of underlying cause (n = 11), CNS relapse (n = 10), secondary intracerebral tumors (n = 3), posterior reversible encephalopathy syndrome (n = 4), seizures (n = 3), migraine (n = 2), degenerative Parkinson-like syndrome (n = 1) or neurological and psychiatric syndromes without detection of a clear etiology (n = 5). Median time between HCT and onset of CNS symptoms was 4.6 months (range, 0-155 months). Treatment was guided by the respective diagnosis and included systemic or intrathecal chemotherapy, anti-infective or anti-inflammatory drugs, anticoagulation and recanalization leading to resolution of neurological symptoms (n = 31), amelioration (n = 15), stabilization (n = 28) or disease progression and death (n = 28). Cumulative incidence of CNS disease-related deaths was 11% and 21% at 100 days and 12 months, respectively (1% and 2% for the complete patient cohort of 1,204 patients). Median overall survival was 13.8 and 6.4 months after HCT and onset of CNS complications, respectively. Conclusions: CNS manifestations account for a significant proportion of HCT-related complications resulting in significant mortality and morbidity with 70% patients experiencing permanent neurological damage. Therefore, precise and prompt diagnosis is essential to guide therapy and can be sometimes challenging. Disclosure: No conflict of interest disclosed. P265

Antibody protection against measles, mumps and rubella depends on underlying disease and conditioning regimen in patients given allogeneic hematopoietic cell transplantation Bögeholz J.1, Manz M.G.1, Schanz U.1, Müller A.M.S.1 Universitätsspital Zürich, Division of Hematology, Zürich, Switzerland

Introduction: Following allogeneic hematopoietic cell transplantation (HCT) the immune system is severely compromised due to pharmacological immunosuppression and disruption of lymphoid tissues by conditioning and donor T cell alloreactivity. Guidelines recommend re-vaccinations against certain pathogens, however, live-vaccines should be avoided in the early post-HCT period. Here, we studied the dynamics of antibody (AB) titers against measles, mumps, and rubella (MMR) during the first 3 years (y) post-HCT. Methods: We retrospectively analyzed serial AB titers in 79 patients who underwent allogenic HCT from HLA-matched related donors from 20092014 at our center. AB titers against MMR were measured prior to HCT, at 6 months (m), 1, 2, and 3y post-HCT. Results: Most patients had protective AB titers (measles 94%, mumps 85%, rubella 92%) prior to HCT. Although only recipients of seropositive donors were included in the analysis AB protection was lost in a substantial proportion of patients post-HCT (protection @2y in 79% for measles, 50% for mumps 50%, 76% for rubella). The proportion of patients who retained protective AB titers at 1y post-HCT was higher in recipients of mobilized peripheral blood compared with bone marrow (BM) grafts (measles p = 0.01, mumps = 0.02, rubella p = 0.13). For rubella, absolute AB titers were available. Figure 1 displays the decline of rubella AB concentrations @6m, 1y, 2y, and 3y post-HCT. Patients with lymphoid malignancies, MAC regimen, ongoing GVHD and pharmacological immunosuppression had a steeper decline of rubella AB titers.

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Fig. 1.

Conclusion: Despite donor seropositivity there is a marked decline of AB titers post-HCT with loss of protection in a substantial proportion of patient. Surprisingly, BM grafts did not provide better AB protection post-HCT, despite their higher content of plasma cells. Together with the observations that (i) patients with lymphoid malignancies (who have received (B-) lymphocyte targeted therapies prior to HCT) had lower AB levels, while (ii) those given RIC have higher AB levels our data suggest that residual host plasma cells significantly contribute to AB production during the first years post-HCT. Disclosure: No conflict of interest disclosed. P266

Allogeneic stem cell transplantation for secondary and therapy-related acute myeloid leukemia: a single center analysis of long-term outcome Hemmati P.1, Pfeifer K.1, Vuong L.1, Jehn C.1, Terwey T.1, le Coutre P.1, Dörken B.1, Arnold R.1 Charité – Universitätsmedizin Berlin, Medizinische Klinik m.S. Hämatologie, Onkologie und Tumorimmunologe, Berlin, Germany 1

Introduction: Acute myeloid leukemia (AML) evolving from myelodysplastic syndrome (MDS) or myeloproliferative neoplasia (MPN) as well as therapy-related AML represent distinct entities in the current WHO classification and are characterized by an inferior outcome compared to de novo AML. This is mainly due to unfavorable cytogenetics, older age and/or comorbidities as well as poor response to induction therapy. For the majority of patients, allogeneic stem cell transplantation represents the only option to achieve long-term disease control and definitive cure. Patients and methods: With regards to long-term outcome, we retrospectively analyzed 204 patients with secondary AML, i.e. AML following MDS/MPN, therapy-related AML, or MDS RAEB-II transplanted at our center between 1996 and 2015. Cytogenetic risk was categorized according to the SWOG/ECOG classification and was favorable (N = 3; 2%), intermediate (N = 94; 46%), unfavorable (N = 84; 41%), or unknown/undetermined (N = 23; 11%). At the time of alloSCT, 98 patients (48%) were in complete hematologic remission (CHR), whereas 106 patients (52%) had active disease. Standard myeloablative conditioning (MAC) was used in 41 patients (20%), whereas reduced intensity conditioning (RIC) was applied in 163 patients (80%). Grafts were from related (N = 51; 25%) or unrelated (matched: N = 112; 55% or mismatched: N = 41; 20%) donors. Results: The median follow-up of the surviving patients was 46 (5-24) months. At 1, 3, 5, and 10 years after alloSCT, disease-free survival (DFS)

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of the entire cohort was 50%, 38%, 36%, and 27%. At the same time points, the cumulative incidence of relapse (CI-R) or non-relapse mortality (CINRM) was 30%, 37%, 37%, and 40% or 20%, 25%, 27%, and 33%, respectively. Univariate analyses revealed a number of factors associated with inferior outcome, e.g. poor-risk cytogenetics, the presence of tAML, the use of reduced-intensity conditioning, and comorbidities, whereas age, donor type (unrelated versus unrelated), and remission status had no significant impact on overall results. Conclusions: Our results indicate that alloSCT represents an important treatment option for patients with secondary and therapy-related AML. However, relapse rates approach 40% at 10 years, which prompts the development of novel strategies to prevent disease recurrence. Furthermore, NRM remains high. Therefore, patients must be carefully selected for transplantation. Disclosure: No conflict of interest disclosed. P267

BKV disease during the first 100 days after allogeneic hematopoietic stem cell transplantation – results of a screening program and retrospective analysis Posdzich P.1, Herling M.1, Chemnitz J.1, Leitzke S.1, Di Christanziano V.2, Höller K.2, Kaiser R.2, Scheid C.1, Holtick U.1 Klinik I für Innere Medizin, Universität Köln, Köln, Germany, 2Institut für Virologie, Universität Köln, Köln, Germany 1

Introduction: BK virus (BKV) is a human polyomavirus often acquired in childhood which can reactivate after allogeneic hematopoietic stem cell transplantation (HSCT). BKV reactivation may present in different ways from asymptomatic viruria to hemorrhagic cystitis or nephritis. Methods: We evaluated 377 patients (159 male, 218 female, median age 52 years) who were transplanted at the University of Cologne between 2008 and 2013 to assess incidence and risk factors for BKV disease after allogeneic HSCT. All 377 patients were screened routinely for BKV in urine and serum. We defined BKV disease as hemorrhagic cystitis (HC) with BK virus >100.000/ml in urine, hematuria (2+ until 4+ in dipstick test) and negative urine microbiology. To find risk factors for BKV disease, the impact of conditioning, CMV status, graft-versus-host disease (GvHD), underlying disease, donor mismatch and stem cell source was analyzed. Results: According to our definition, BKV cystitis occurred in 37 of 377 patients (9,8%) in the first 100 days after HSCT. In 14 of these 37 patients (37,8%) BKV was also detected in serum. BKV was detected in the urine of 194 patients (51,4%) at any time during the screening program, but only 19,1% developed hemorrhagic cystitis. Sixty-two percent of patients suffering from BKV disease were female, 37,8% male. BKV disease was diagnosed in 13,3% of patients not in complete remission as opposed to 5,4% of patients in complete remission prior to conditioning (p = 0,010). BKV disease was less common in matched related or unrelated (7,7%) as compared to mismatched or haplo-identical donors (16,7%; p = 0,012). Acute GvHD grade II to IV was described in 122 of 377 patients (32,36%). Of them, 13,9% suffered from BKV cystitis in comparison to 7,84% with no or grade I acute GvHD, which did not reach statistical significance. We could not detect any correlation between BKV disease and administration of cyclophosphamide, total body irradiation or anti-thymocyte globuline. There was no association with the graft source. BKV disease was no predictor for non-relapse mortality, overall and relapse-free survival. Conclusion: BKV disease is a common complication after allogeneic HSCT. The virus can be found in the urine of about half of our transplanted patients, but only a small proportion developed an HC. State of remission before conditioning and donor mismatch are associated with the development of BKV disease, which had no impact on survival in our cohort.

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High CD34+ cell dose is associated with increased early mortality after allogeneic hematopoietic stem cell transplantation Neumann K.1, Lange S.1, Kragl B.1, Glass Ä.2, Große-Thie C.1, Wittke C.1, Freitag S.1, Henze L.1, Kleine H.-D.3, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany, 2Institute for Biostatistics and Informatics in Medicine and Ageing Research, Rostock, Germany, 3Seracell Stammzelltechnologie GmbH, Rostock, Germany 1

Introduction: Survival after allogeneic hematopoietic stem cell transplantation (HSCT) is influenced by a variety of recipient- and graft-related factors as well as conditioning regimen. Whereas some research groups point out higher mortality rates with increased CD34+ cell doses, others do not. We aimed at determining the impact of CD34+ cell dose on early mortality after allogeneic HSCT in a retrospective cohort analysis. Patients and methods: Data sets of 436 consecutive patients who underwent allogeneic HSCT at our center from 1998 to 10/2015 were evaluated regarding their early mortality rates (d+30, d+60). Cox regression analysis comprised the variables viable CD34+ cell dose, recipient sex, age, body mass index (BMI), HSCT indication, conditioning intensity, stem cell source, post-transplant immunosuppression and donor type. Results: Median recipient age at time of HSCT was 49 (15-76) years. Indications for HSCT comprised AML/MDS (44.5%), ALL (8.3%), CML (10.1%), NHL (15.1%), CLL (4.4%), Hodgkin’s disease (0.7%), multiple myeloma (8.0%) and other diseases (8.9%). Conditioning was carried out employing myeloablative (27.6%) and reduced intensity or non-myeloablative (72.4%) regimens with treosulfan/fludarabine-based conditioning being most common (53.2%). Matched unrelated donors (54.2%) and matched related donors (33.1%) accounted for the most frequent donor types. Patients received a median CD34+ cell dose of 4.4×106/kg (0.4×10622×106/kg). Grafts originating from bone marrow (n = 72) and peripheral blood (n = 364) contained a median CD34+ cell dose of 2.0×106/kg and 5.0×106/kg (p < 0.001). Mortality at d+30 and d+60 amounted to 6.9% and 10.7%, respectively. In multivariable analysis, cell numbers < 2.0×106 (14.0%) and even < 1.0×106 CD34+ cells/kg (2.3%) were not found to negatively impact early mortality. Conversely, patients receiving ≥9.0×106 CD34+ cells/kg (7.1%) experienced significantly increased mortality at d+30 (p = 0.027) and d+60 (p = 0.041). Additionally, myeloablative conditioning proved to be associated with significantly higher early mortality compared to reduced intensity or non-myeloablative regimens (d+30: p = 0.005; d+60: p = 0.037). Conclusions: Early post-transplant mortality seems to be increased following allogeneic HSCT of viable CD34+ cell doses ≥9.0×106 CD34+ cells/ kg. Disclosure: No conflict of interest disclosed. P269

Overall and median survival of patients after allogenic stem cell transplantation with conditioning by treosulfan and fludarabin Freitag S.1, Kragl B.1, Wittke C.1, Brueckner F.1, Gläser D.1,2, Große-Thie C.1, Henze L.1, Hilgendorf I.1,3, Gläser H.1, Freund M.1, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany, 2Clinic of Internal Medicine III – Hematology/Oncology/Hemostaseology, Palliative Care, Complementary Medicine, Klinikum Südstadt Rostock, Rostock, Germany, 3 Clinic of Internal Medicine II – Division of Hematology and Internal Oncology, Jena University Hospital, Jena, Germany 1

Disclosure: No conflict of interest disclosed.

Introduction: Allogeneic stem cell transplantation is frequently applied in variable hematological diseases. During the last decade applied conditioning regimens have shifted from myeloablative to non-myeloablative and reduced intensity conditioning regimen (RIC). Treosulfan (Treo) and

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fludarabine (Flud) + antithymocyte globulin (ATG) based regimens have been evaluated successfully in several phase-II- studies. One large phaseIII-study in AML is ongoing. Here we report on a large cohort of stem cell transplantation (SCT) recipients that received Treo/Flud conditioning before SCT. Method: We analyzed consecutive allo-SCT patients in the time period from 2/1998 to 4/ 2016 who received conditioning therapy using Treo/ Flud based regimens in regards to patient characteristics and overall survival. Results: A total of 240 patients were enrolled. The patient population consisted of 137 (57%) males and 103 (43%) females with a median age of 52 years (range: 16-76 years). Underlying diseases were: acute leukemia 99 (41%), lymphoma 42 (17%), chronic leukemia 38 (16%), plasmacell disorders 22 (9%), MDS 20 (8%), and other 19 (8%). Total Treo doses were median 42g/m² (30g/m²-50g/m²) usually split in 3-5 single doses on d-6 to d-2. Flud total dose was in general 150/m² divided onto 5 days. GVHD prophylaxis was in general cyclosporine A/ MTX based. Unrelated alloSCT recipients received in addition antithymoglobulin. Most transplants were matched related (71/29%) and matched unrelated (163/68%). Early mortality rates were as follows: 30d: 12 (5%), 60d: 23 (9%), 100d: 39 (16%). The 1 year and 5 years survival rates were 62% and 45% respectively. The median overall survival for all patients was 589 days (range: 10- 5901d). Overall survival varied based on underlying disease status. Conclusion: In our large but heterogenous cohort Treo/Flud based conditioning is associated with moderate early mortality rates compared to historical myeloablative conditioning regimen. Disclosure: No conflict of interest disclosed. P270

Are the polyomaviruses BK and JC associated with opportunistic infections, graft versus host disease or worse outcome in adult patients receiving their first allogenic stem cell transplantation with low dose alemtuzumab? Schneidewind L.1, Neumann T.2, Knoll F.2, Zimmermann K.3, Smola S.1, Krüger W.2 Universitätsklinikum des Saarlandes, Institut für Virologie, Homburg/ Saar, Germany, 2Universitätsmedizin Greifswald, Klinik für Innere Medizin C, Hämatologie/Onkologie, Greifswald, Germany, 3Universitätsmedizin Greifswald, Friedrich Löffler Institut für Medizinische Mikrobiologie, Greifswald, Germany 1

Introduction: Polyomaviruses BK (BKPyV) and particularly also JC (BKPyV) are important pathogens in allogenic stem cell transplantation. Their association with other opportunistic infections like CMV reactivation and acute or chronic GvHD is controversial discussed. However, there is a lack of data for adult allogenic stem cell transplantation and especially for JCPyV. We conducted a retrospective study of adult patients receiving their first allogenic stem cell transplantation with low dose alemtuzumab at the University Hospital Greifswald. Methods: Data from all patient records (64) of adult patients undergoing their first allogenic stem cell transplantation and receiving low dose alemtuzumab for GvHD prophylaxis from 03/2010, when quantitative PCR for the BKPyV and JCPyV was established, to 12/2014 were collected including outpatient data of our clinic. Follow up time was two years after infusion of the stem cell product. Results: We found acute leukaemia being the most frequent underlying disease (45.3%), conditioning included myeloablative (67.2%) and non-myeloablative protocols (32.8%). All patients received alemtuzumab 10 mg at day -2 (respectively 20 mg in case of mismatch) as GvHD prophylaxis before transplantation. On the whole, infectious complications occurred rarely when receiving low dose alemtuzumab, e. g. BKPyV associated hemorrhagic cystitis was diagnosed in 10 patients (15.6%). There was no significant association of BKPyV or JCPyV with CMV reactivation, EBV reactivation, HHV-6 or parvovirus B19 infection, if they required treatment. Interestingly, there was a significant association of BKPyV associated hemorrhagic cystitis with toxoplasmosis (p = 0.013). Additionally, there was a significant link with BKPy and JCPy viruria at the

Oncol Res Treat 2016;39(suppl 3):1–104

same time point with toxoplasmosis (p = 0.047). In this study BKPyV and JCPyV were not associated with acute or chronic GvHD. Furthermore, BKPyV or JCPyV were not associated with relapse or death. Conclusion: In our study of adult allogenic stem cell transplant recipients receiving low dose alemtuzumab for GvHD prophylaxis we found a low frequency of viral reactivations and no association of BKPyV or JCPyV with severe viral infections or GvHD. Only the association of both polyomaviruses with toxoplasmosis was significant. This is a novel and interesting finding. Prospective studies with drug monitoring and monitoring of the immune system are necessary to confirm these findings for this kind of population. Disclosure: Laila Schneidewind: Expert Testimony: Autorin wird aktuell mit einem Forschungsstipendium der Monika Kutzner Stuftung unterstützt. William Krüger: No conflict of interest disclosed. P271

Second allogeneic hematopoietic cell transplantation to treat relapsed acute leukemia Hagmaier V.1, Schneidawind C.1, Duerr-Stoerzer S.1, Faul C.1, Kanz L.1, Bethge W.1, Schneidawind D.1 Department of Medicine II, Eberhard Karls University, Tübingen, Germany

Introduction: Allogeneic hematopoietic cell transplantation (HCT) is the treatment of choice for high-risk myeloid and lymphoid leukemias. Relapse after allogeneic HCT is associated with a dismal prognosis and further therapeutic options are limited. One potential curative approach is a second allogeneic HCT. However, there is no consensus about optimal transplant modalities and suitable patients. Methods: We performed a retrospective analysis of our institutional database to evaluate risk factors that influence survival after a second allogeneic HCT for the treatment of relapsed acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). Results: We identified 40 patients (f = 13, m = 27) with a median age of 41 years (range, 18-65) that received a second allogeneic HCT to treat relapsed AML (n = 29) or ALL (n = 11) at our institution from 2002 to 2015. At time of second HCT, 42% of patients were in complete remission (CR). Grafts from either matched related (MRD, n = 3), matched unrelated (MUD, n = 12), mismatched related (MMRD, n = 22) or mismatched unrelated donors (MMUD, n = 3) were used. 3 patients received an allograft from the same donor as at primary HCT. Current overall survival (OS) is 15/40 patients with a median follow-up of 8 months (range 1-156) of patients alive resulting in a Kaplan-Meier estimated 3-year event-free survival (EFS) and OS of 33%. Causes of death were relapse (n = 13), infections (n = 7) and others (n = 5). Cumulative incidence of non-relapse mortality (NRM) and relapse at 3 years was 29% and 44%, respectively. Early transplantation within one year after the previous allogeneic HCT showed a similar outcome compared with later HCT (3-year OS 30% vs. 33%, p = 0.41). Likewise, relapse within 6 months after the first allogeneic HCT was not associated with a significantly reduced 3-year OS after the second allogeneic HCT (30% vs. 35%, p = 0.33). However, ALL patients showed a decreased 3-year OS (13% vs. 39%, p = 0.05). Moreover, we found that a second allogeneic HCT in CR translated into a significantly improved 3-year OS when compared to patients with active disease (51% vs. 21%, p = 0.02). Conclusions: A second allogeneic HCT is a curative treatment option for relapsed acute leukemias in selected patients. Prior salvage chemotherapy resulting in CR is an important prerequisite for long-term disease-free survival. Disclosure: No conflict of interest disclosed.

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Posterdiskussion

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Immuntherapie

Ex vivo analysis of the tumor antigen-specific T cell repertoire in a melanoma patient

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Lennerz V.1, Schroers B.1, Luebcke S.1, Fatho M.1, Kukla K.1, Brettschneider J.1, Woelfel C.1, Pagel S.1, Zhao F.2, Echchannaoui H.1, Theobald M.1, Schadendorf D.2, Paschen A.2, Woelfel T.1

Fluorescence based reporter cells rapidly identify and distinguish functional Chimeric Antigen Receptors (CARs) Rydzek J.1, Jutz S.2, Leitner J.2, Wallstabe L.1, Nerreter T.1, Huppa J.B.3, Einsele H.1, Steinberger P.2, Hudecek M.1 Universitätsklinikum Würzburg, Medizinische Klinik II, Hämatologie, Würzburg, Germany, 2Medizinische Universität Wien, Institut für Immunologie, Zentrum für Pathophysiologie, Infektiologie und Immunologie, Wien, Austria, 3Medizinische Universität Wien, Institut für Hygiene und Angewandte Immunologie, Zentrum für Pathophysiologie, Infektiologie und Immunologie, Wien, Austria 1

Introduction: Adoptive immunotherapy with chimeric antigen receptor (CAR)modified Tcells is under intense preclinical and clinical investigation involving a rapidly increasing spectrum of new CAR designs and target antigens. Here, we present a novel reporter cell line that enables highthroughput testing and identification of functional receptor designs to inform the selection of CARs for indepth preclinical validation and clinical translation. Methods: Triple parameter reporter (TPR) cells were derived from Jurkat cells and modified with three inducible reporter genes, each expressing a distinct fluorophore under control of the Tcell transcription factors NFκB, NFAT or AP1. TPR cells were transduced with CAR constructs, cocultured with target antigenexpressing stimulator cells and reporter geneactivation was analyzed by flow cytometry. Stimulator cells with membranebound antiCD3 scFv that triggers CD3 on TPRs were used as positive control and reference. Results: We transduced TPR cells with CD19 and ROR1specific CARs that contained a signaling module of CD3zeta and 41BB (2nd generation CAR) and cocultured them with CD19 and ROR1expressing stimulator cells respectively. We detected specific and rapid reporter gene induction mediated by NFκB and NFAT as early as 9 hours and peaking between 24 to 48 hours of coculture. We then compared the activation signal that was obtained with CARs incorporating CD28 or 41BB alone (2nd generation CAR) or in tandem (3rd generation CAR). Interestingly, we detected stronger induction of NFκB in CARs providing 41BB rather than CD28 costimulation at 24 hours. However, the inclusion of tandem CD28 and 41BB costimulation in 3rd generation CARs did not result in stronger or more durable activation, consistent with our observation that primary Tcells expressing the 3rd generation CD19 or ROR1CAR did not confer superior effector functions in vitro and in vivo. Conclusions: Our data demonstrate the potential of TPR cells to evaluate CARs based on the induction of key Tcell transcription factors. We are currently integrating TPRs into the testing of novel CAR designs with enhanced stimulatory and costimulatory capacity to rapidly identify the most potent formats from a large pool of constructs. We are confident our TPR test system will accelerate preclinical development and facilitate clinical translation of CARs with optimal antitumor function and safety profile for hematologic and solid malignancies. Disclosure: No conflict of interest disclosed.

University Medical Center (UMC) Mainz, Medical Department 3, University Cancer Center (UCT), Research Center for Immuntherapy (FZI) of the Johannes Gutenberg-University and German Cancer Research Consortium (DKTK), partner site Frankfurt/Mainz, Mainz, Germany, 2University Hospital, University Duisburg/Essen, Department of Dermatology, and German Cancer Research Consortium (DKTK), partner site Essen/Düsseldorf, Essen, Germany 1

Introduction: In patient Ma-Mel-86 surviving metastatic disease for approximately seven years, four melanoma lines were derived from distinct lymph node metastases occurring in the years 1, 3, 4 and 7. The melanoma lines exhibited various immune escape phenotypes including partial or complete HLA class I deficiency. Blood-derived melanoma-reactive CD8+ abTCR+ T cell clones were found to recognize more than seven non-mutated shared or mutated neoantigens in HLA-restricted and more than 2 surface membrane proteins in HLA-independent manner. In a next step, we have investigated how the melanoma-reactive T cell repertoire evolved during the course of the disease. Methods: The TCRVb chains of 18 T cell clones with known and unknown specificities were sequenced and served as target sequences. Each 3.6 µg of genomic DNA (~ 5.5×10e5 cells) from 2 PBMC samples collected in year 1 (sample 1) and year 3 (sample 2) were subjected to TCRVb high-throughput sequencing using the ImmunoSEQ Assay (Adaptive Biotechnologies, Seattle, USA). Sample 2 had been collected six months after the first of two HLA class I-negative lymph node metastases had emerged. Results: Of 18 target TCRVb sequences, only 2 were found in sample 1. They were expressed by HLA-restricted T cell clones recognizing a mutated neoantigen and a yet unknown antigen, respectively. Both TCR were also detectable in sample 2 indicating that they were expressed by memory T cells. In sample 2, altogether 15/18 target TCRVb sequences were detectable. Among these were 6 TCRVb sequences assigned to HLA-independent T cells. TCRs against known HLA-restricted mutated and shared antigens were detected at low (< 0,01%) to intermediate (0,01-0,1%) frequencies, whereas HLA-independent TCRs were detected at intermediate to high frequencies (>1%). Only the above mentioned HLA-restricted TCR against a yet unknown antigen was detectable at high frequencies in both samples. Conclusions: With disease progression, the patient’s anti-melanoma T cell repertoire became considerably broader. Two TCRs were assigned to long-term memory T cell responses. Our data indicate that, as soon as HLA-deficient metastases appeared, a dominant HLA-independent anti-melanoma T cell repertoire evolved. Disclosure: No conflict of interest disclosed. P274

Immunization with a solid nanoscopic imiquimod suspension enhances induction of CTL responses Aranda Lopez P.C.1, Denny M.2, Hartmann A.-K.3, von Stebut-Borschitz E.4, Stassen M.3, Schild H.3, Theobald M.1, Langguth P.2, Radsak M.P.1 Johannes Gutenberg University Medical Center, Third Department of Medicine, Mainz, Germany, 2Johannes Gutenberg University, Biopharmaceutics and Pharmaceutical Technology, Mainz, Germany, 3Johannes Gutenberg University Medical Center, Institute for Immunology, Mainz, Germany, 4Johannes Gutenberg University Medical Center, Dermatology, Mainz, Germany 1

Introduction: Transcutaneous immunization (TCI) is a novel vaccination strategy to induce strong therapeutic cytotoxic T-lymphocyte (CTL) responses by directly targeting skin-resident professional antigen-presenting cells (APC). This vaccination approach is very promising to overcome current limitations of standard vaccination approaches that are mostly effective in prophylaxis, but not in the treatment of diseases. In this con-

Abstracts

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text, we have developed a TCI method based on a synthetic TLR7 agonist imiquimod. We generated a freeze-dried solid suspension of crystalline imiquimod (IMI-Sol), which upon shear force (i.e. spreading on the skin) transforms to a liquid phase that permeates into the skin. Methods: Mice were shaved on their backs and immunized on two consecutive days either with the commercially available Aldara® together with SIINFEKL peptide or with IMI-Sol followed by application of SIINFEKL in officinal cremor basalis. Induction of immune response was characterized by analysis of peptide-specific T-cells and in vivo cytolytic activity. Concerning the signalling cascade we immunized transgenic mouse strains, deficient for various signalling molecules as well as mastcell-deficient mice. To evaluate inflammation in the skin, we treated mice either two times (standard protocol) or four times (psoriasis like protocol) and measured epidermal thickness compared to untreated controls. Results: Comparing IMI-Sol vaccination with the commercially available Aldara® we found superior vaccination capacity of IMI-Sol. Using TLR7/or MyD88-/- mice, we confirmed that the induction of CTL responses with IMI-Sol was completely dependent on the TLR/MyD88 pathway. In contrast to Aldara® treatment, CTL responses induced by IMI-Sol are mastcell as well as IL-1 receptor independent. Analysing inflammation in the skin revealed increased epidermal thickness after IMI-Sol using our standard protocol (2 treatments), whereas comparable skin reactions were observed in a psoriasis like experiment (4 treatments). Conclusions: All together the development of revised preparations and the deeper understanding of the underlying mechanisms may harbour the key for the rationale design of a next generation transcutaneous vaccination platform that can be used for the treatment of persistent infections and cancer. Disclosure: No conflict of interest disclosed. P275

Construction of an HLA-independent single-chain T cell receptor (scTCR) against TRP2 Wölfel M.1, Wölfel C.1,2,3, Paschen A.4,5, Wölfel T.1,3,6, Theobald M.1,2,3, Echchannaoui H.1,2,3 University Medical Center (UMC) of the Johannes Gutenberg-University, Internal Medicine III, Mainz, Germany, 2University Cancer Center (UCT), Mainz, Germany, 3German Cancer Research Consortium (DKTK), partner site Frankfurt/Mainz, Mainz, Germany, 4University Hospital, University Duisburg/ Essen, Dermatology, Essen, Germany, 5German Cancer Research Consortium (DKTK), partner site Essen/Düsseldorf, Essen, Germany, 6Research Center for Immunotherapy (FZI), Mainz, Germany 1

Introduction: From the peripheral blood of melanoma patient Ma-Mel-86 with clinically evident HLA I- metastases we have isolated HLA-independent CD8+ αβT cell receptor (TCR)+ cytolytic T cells recognizing TRP2 as an intact molecule on the cell surface of autologous melanoma cells. Their TCR chains have been cloned and proved to be functional in a retroviral bicistronic double-chain construct (dcTCR). We intended to design a single-chain (sc)TCR construct to reduce the risk of mispairing with endogenous TCRs upon adoptive transfer and thereby enhance both safety and efficacy. Methods: The scTCR was constructed in analogy to the procedure described by Knies et al. (Oncotarget 7:21119, 2016). It consists of the TRP2-recognizing variable (V) domains of the original dcTCR covalently bridged with a 19mer Glycine/Serine-rich linker (Li) and murine (mu) constant α and β domains (Vα-Li-Vβ-muCβ-muCα). Non-native cysteine residues were introduced into the murine C regions to generate a second disulfide bond between the C regions and thus minimize mispairing with endogenous TCRs. Finally the scTCR construct was cloned into retroviral vector pMX-DEST via Gateway recombination. Functional testing was performed with IFNγ ELISPOT and standard 51Cr release assays. Results: Jurkat-76 cells lacking endogenous TCR and peripheral blood lymphocytes (PBL) from healthy donors were transduced with pMX constructs encoding the anti-TRP2 dcTCR and the anti-TRP2 scTCR. scTCR and dcTCR surface expression was confirmed by flow cytometry with an

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antibody against the murine αβTCR constant region. Successfully transduced PBL were expanded by weekly restimulations with TRP2+ MaMel-86 cells. Both CD4+ and CD8+ T cell subsets expressing the scTCR recognized Ma-Mel-86 cells as well as HEK293Tcells that were HLAI/IIdue to TALEN-mediated knockout of the genes encoding B2M (ß2 microglobulin) and CIITA (HLA class II transactivator) and that were transfected with titrated amounts of TRP2-encoding cDNA. In comparative functional analyses the scTCR and the dcTCR constructs were equivalent and recognized both murine and human TRP2. Conclusions: The anti-TRP2 scTCR construct specifically recognized TRP2 in an HLA-independent fashion. It was equivalent to the parental dcTCR construct in ELISPOT and lysis assays. With respect to potential therapeutic applications of HLA-independent TCR against tumor-associated surface molecules, scTCR constructs form a basis for versatile and modular applications. Disclosure: No conflict of interest disclosed. P276

Glyco- and Fc engineering of IgA antibodies for cancer immunotherapy improved pharmacokinetics & myeloid effector cell engagement Kretschmer A.1, Lohse S.2, Meyer S.3, Meulenbroek L.A.P.M.3, Jansen J.H.M.3, Möginger U.4, Sondermann P.5, Kolarich D.4, Leusen J.H.W.3, Valerius T.1 Division of Stem Cell Transplantation and Immunotherapy, 2nd Department of Medicine, UKSH and CAU, Kiel, Germany, 2Institute of Virology, Saarland University Medical Center and Saarland University Faculty of Medicine, Homburg/Saar, Germany, 3Laboratory for Translational Immunology, University Medical Center Utrecht, Utrecht, Netherlands, 4Department of Biomolecular Systems, MPI for Colloids and Interfaces, Potsdam, Germany, 5SppreMol GmbH, Martinsried, Germany 1

Introduction: Antibodies of IgA isotype play an important role in bridging adaptive and innate immunity. FcαRI (CD89) - dependent engagement of myeloid cells appears to be crucial to activate effector mechanisms like phagocytosis or antibody-dependent cell-mediated cytotoxicity (ADCC). Recently, there has been increasing evidence that myeloid cells constitute important effector cells in cancer and cancer immunotherapy. Thus, we decided to develop production and purification technologies for recombinant IgA antibodies, which demonstrated in vivo efficacy in syngeneic and xenogeneic tumor models. Here, we describe an Fc engineering approach to further improve their immunotherapeutic potential. Methods: Recombinant IgA antibodies against the epidermal growth factor receptor (EGFR) were produced by co-transfecting CHO-K1 cells with vectors encoding the variable region of Cetuximab and Igα heavy and κ light chain constant regions, respectively. An Fc engineered IgA2m(1) antibody was generated by mutating two N-glycosylation sites (166 and 337) and by removing two free cysteines (311 and 471). The resulting antibody variant was compared to wild type IgA2 regarding biochemical characteristics as well as Fab- and Fc- mediated effector functions. Additionally, serum half-life and in vivo efficacy in a xenogeneic FcαRI- transgenic tumor model were evaluated. Results: Rational engineering of the constant regions of an IgA2m(1) antibody resulted in monomeric IgA molecules with improved biochemical characteristics, identical Fab- and Fc- mediated effector functions, but with significantly lower levels of terminal galactose. This molecule demonstrated lower asialoglycoprotein-receptor (ASGPR) binding and subsequently improved pharmacokinetics in mice. Compared to wild type IgA, this novel molecule displayed enhanced therapeutic efficacy in different in vivo models, which required human FcαRI-dependent myeloid effector cell engagement. Conclusion: These results demonstrated that glyco- and Fc engineering of antibodies from the IgA isotype directed against EGFR leads to an improvement of their immunotherapeutic efficacy by overcoming of some limitations of wild type IgA antibodies (e.g. stability, pharmacokinetics, in vivo efficacy). Thus, these results promote the concept of FcαRI- de-

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pendent engagement of myeloid effector cells as a promising approach for antibody-based tumor immunotherapy. Disclosure: No conflict of interest disclosed. P277

Targeting cancer metabolism for the improvement of checkpoint therapy Thiel A.1, Singer K.1, Renner-Sattler K.1, Kreutz M.1 Universitätsklinikum Regensburg, Medizinische Klinik lll, Regensburg, Germany, 1

A major breakthrough in cancer immunotherapy was the discovery of immune checkpoint proteins and the development of specific inhibitors. Especially blocking PD-1 and PD-L1 has been shown to be associated with an enhanced overall survival in metastatic disease of various tumour entities. However only a limited patient cohort shows sufficient response to the therapy. Therapy failure does not inevitably correlate with the threshold of PD-L1 expression on the tumour in the non-responding patient-population. The reasons for the low response rates need to be elucidated. It has been demonstrated that tumour metabolites, in particular lactic acid suppress the anti-tumor immune response and moreover can skew immune cells to a regulatory phenotype. To investigate the impact of lactic acid and regulatory immune cells on checkpoint inhibition we established an in-vitro co-culture model of B16-F10 melanoma cells and 2C TCR transgenic T cells. The B16-F10 melanoma cells were transduced to express the K(b)-binding peptide SIYRYYGL which is recognized by high-affinity 2C TCR transgenic T cells. We herein demonstrate the differential impact of lactic acid and regulatory immune populations on checkpoint inhibition with anti-PD-1 and anti-PD-L1 in-vitro. The overall objective of our work is the development of combination therapies targeting cancer metabolism for improvement of checkpoint therapy. Disclosure: No conflict of interest disclosed. P279

Glykolipid antigen-specific invariant natural killer T cells for protection from graft-versus-host disease Schmid H.1, Duerr-Stoerzer S.1, Schneidawind C.1, Kanz L.1, Salih H.1, Savage P.2, Schneidawind D.1 Department of Medicine II, Eberhard Karls University, Tübingen, Germany, Department of Chemistry and Biochemistry, Brigham Young University, Provo, United States 1 2

Introduction: Invariant natural killer T (iNKT) cells are potent regulators of immune responses in both humans (TCRα Vα24-Jα18) and mice (TCRα Vα14-Jα18). This particular thymus-derived T-cell subset is restricted to the MHC-I-like and lipid antigen-presenting molecule CD1d. Glycolipids such as α-GalCer induce a strong activation and proliferation of iNKT cells through T-cell receptor signaling. We recently showed in murine studies that adoptively transferred iNKT cells of either donor or third party mice protect from lethal graft-versus-host disease (GVHD) through a robust expansion of donor CD4+FoxP3+ regulatory T cells (Tregs) while preserving graft-versus-tumor (GVT) effects. Immune polarization of iNKT cells is critical for their immunoregulatory function upon adoptive transfer. Since iNKT cells constitute less than 0.5% of human peripheral blood mononuclear cells (PBMCs), in vitro expansion with their glycolipid ligands is required before these cells can be used for cytotherapy. Methods: We tested the impact of three different glycolipids (α-GalCer, PBS57, PBS44) on human iNKT-cell proliferation, subset expansion and immune polarization by flow cytometry, intracellular cytokine staining and ELISA.

Abstracts

Results: Three weeks of cell culture and autologous restimulation with either α-GalCer, PBS57 or PBS44 resulted in a robust proliferation of iNKT cells from human PBMCs. We did not find significant differences in iNKT-cell expansion (1500x vs. 1727x vs. 1772x, p = 0.49) and purity (33% vs. 38% vs. 39%, p = 0.24) with the three different glycolipids tested in our study. Importantly, we observed a growth advantage of the CD4+ subset resulting in an outgrowth from 14% (day 0) to 87% (day 21) of all iNKT cells (p < 0.001). IL-4 secretion of culture expanded human iNKT cells was comparable in the three groups (18% vs. 24% vs. 31%, p = 0.68). Conclusions: All glycolipids induce a robust proliferation of human iNKT cells in vitro. Interestingly, we found a preferential expansion of CD4+ iNKT cells secreting the Th2 cytokine IL-4. This is critical since our previous murine studies indicate that the CD4 subset and Th2 phenotype are associated with immune tolerance and protection from GVHD through Treg expansion. Disclosure: No conflict of interest disclosed. P280

Influence of cytoreductive and immunmodulatory drugs on antibody-based approaches in AML Krupka C.1,2, Lindl B.1,2, Platzer J.1,2, Brauneck F.1,2, Kischel R.3, Kufer P.3, Lichtenegger F.S.1,2, Köhnke T.1,2, Rothe M.1,2, Deiser K.1,2, Augsberger C.1,2, Altmann T.1,2, Spiekermann K.1,4, Hiddemann W.1,4, Subklewe M.1,2,4 LMU Munich; Klinikum der Universität München, Department of Internal Medicine III, München, Germany, 2Helmholtz Zentrum München, Clinical Cooperation Group Immunotherapy, Munich, Germany, 3AMGEN Research (Munich) GmbH, Munich, Germany, 4German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Heidelberg, Germany 1

Previously, we demonstrated that the CD33/CD3 BiTE® antibody construct AMG 330 is able to recruit residual autologous T cells, resulting in effective cytotoxicity against primary AML cells. Based on these data, a phase I study in relapsed/refractory AML has been initiated (NCT02520427). Clinical experience in ALL has shown a clear correlation of leukemic burden and a cytokine release syndrome (CRS) during treatment with blinatumomab. A cytoreductive phase prior to antibody therapy might be beneficial to reduce the severity of the CRS. Furthermore antibody mediated unwanted toxicity is treated with steroids (dexamethasone, DEX) or tocilizumab (TOC, IL-6R antibody). However, little is known about the relevance of these drugs on effector cell function. Therefore we evaluated the influence of cytoreductive- (azacythidine, AZA; decitabine, DEC; cytarabine, ARA-C; hydroxyurea, HU) and immunmodulatory drugs (DEX, TOC) on antibody-mediated T-cell function. T cells and AML cells were cocultured with or without AMG 330 for 3-7 days. T cells were incubated with the drug prior to coculture (AZA, DEC, ARA-C, HU) or simultaneously added (DEX, TOC). Lysis of AML cells and T-cell proliferation were assessed by flow cytometry. Pretreatment of T cells with ARA-C completely abrogated T-cell function (% lysis: untreated(UT) 98.0 vs 20 µM 4.2) and proliferation. AZA and DEC impaired AMG 330-mediated cytotoxicity (% lysis UT vs AZA at 1, 5, 10 µM: 99.8 vs 99.2 vs 52.1 vs 28.6; UT vs DEC at 0.3, 2, 5 µM: 78.9 vs 57.9 vs 48.6 vs 5.8) and T-cell proliferation in a concentration dependant manner. In contrast, pretreatment with HU had no effect on T-cell function (% lysis: UT vs HU at 10, 100 µM: 100 vs 100 vs 97.3) and proliferation. The addition of DEX to primary AML cultures considerably impaired T-cell proliferation (fold change T cells: UT 12.9 vs DEX 1.3) and IFNγ secretion (pg/ml: UT: 813 vs DEX: 64). TOC had no negative effect on T-cell proliferation (fold change T cells: UT: 5.0 vs TOC: 4.9). Similarly, secretion of IFNγ (pg/ml: UT: 712 vs TOC: 839) was not affected. In summary we could show that in contrast to HU, pretreatment with ARA-C, AZA and DEC negatively influenced T-cell function. Furthermore, in comparison to TOC the use of DEX considerably decreased AMG 330 mediated T-cell proliferation and cytokine secretion. As most antibody based immunotherapeutic strategies depend on effector cell function, our data support the use of HU and TOC for combinatorial approaches.

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Disclosure: Christina Krupka: Expert Testimony: wissenschaftliches Projekt finanziell unterstützt durch AMGEN Research Munich GmbH Marion Subklewe: Advisory Role: AMGEN Research (Munich); Expert Testimony: wissenschaftliches Projekt finanziell unterstützt durch AMGEN Research Munich GmbH

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Mobilization of tissue-resident memory cells by the granulocyte colony-stimulating factor (G-CSF) Olfe L.1, Asadi K.1, Becker S.1, Mensen A.1, Szyska M.2, Tietze-Bürger C.3, Scheibenbogen C.1, Schetelig J.4, Dörken B.5, Arnold R.5, Na I.-K.1,2,5 Institute for Medical Immunology, Charité CVK, Berlin, Germany, 2Experimental and Clinical Research Center (ECRC), Berlin, Germany, 3Charité Stem Cell Facility, Berlin, Germany, 4Universitätsklinikum Carl Gustav Carus, Technische Universität, Dresden, Germany, 5Department of Hematology, Oncology and Tumorimmunology, Charité, Berlin, Germany 1

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Long term efficacy of WT1 peptide vaccination in patients with WT1 expressing AML /MDS and solid malignancies Lukas K.1, Scheibenbogen C.2, Asemissen A.M.3, Busse A.1, Ochsenreither S.1, Blau I.1, Baldus C.1, Thiel E.1, Keilholz U.4, Letsch A.1 Charité, Campus Benjamin Franklin, Med. Klinik III, Berlin, Germany, 2Charité – Universitätsmedizin Berlin (CVK), Institute of Medical Immunology, Berlin, Germany, 3UKE Hamburg, Hematology, Oncology and Bone Marrow Transplantation, Hamburg, Germany, 4Charite-Unversitätsmedizin, Charité Comprehensive Cancer Center, Berlin, Germany 1

Immunotherapy is a novel strategy for generating antitumor immunity as part of cancer treatments. The present study aims to assess the feasibility, immunogenicity and clinical effects of WT1 peptide vaccination based on the growing evidence that Wilms´ tumor protein 1 (WT1) is a promising tumor antigen for the development of universal cancer vaccines. In this light, we conducted two separate phase-II proof of principle vaccine trial in patients with active AML/MDS and with WT1 overexpressing solid tumors. Here we concentrate on patients remaining on vaccination for at least one year in order to analyze characteristics for long-term vaccination success. The vaccination schedule was identical for both trials: 0.2mg HLA-A*0201-restricted WT1.126-134 peptide admixed with 1 mg KLH (day 3), and 62.5µg GM-CSF (day 1-4) every two weeks x 4 s.c./i.d., followed by either biweekly or every four weeks for 10 months and at increased intervals of up to 3-monthly thereafter. The two trials included patients with active AML and high-risk MDS (n = 20), patients with AML in high risk complete remission (n = 9), as well as with mostly heavily pretreated ovarian (n = 7), thyroid (n = 2), breast (n = 1), gastric (n = 1), and larynx cancer (n = 1), astrocytoma (n = 1) and mesothelioma (n = 4). Standard criteria were used for response assessment. Patients remained on study in absence of limiting toxicity and disease progression.Toxicity to treatment was mild , and no patient went off protocol for adverse events or consent withdrawal. Eleven patients remained on study for more than 1 year with disease control, including 3/19 patients with active AML, 4/9 with AML in complete remission, 2/7 patients with ovarian cancer and 2/4 with mesothelioma. Of these 11 patients four remained on study for more than 3 years, including 3 patients with AML (75, 70+ and 114+ months) and one mesothelioma patient (84+ months). We could demonstrate remarkable longterm responses induced by WT1 vaccination in patients with AML and solid tumors. Further vaccine development, especially in combination with checkpoint inhibitors seem promising. Overall, no correlation between prior treatment and duration on vaccine was observed, since 2/4 patients in long-term disease control had previously aggressive chemotherapy. Of the solid tumor patients, chemotherapy-pretreated ovarian cancer and mesothelioma appear to be sensitive to WT1 vaccine, although other histologies cannot be excluded due to low patient numbers. Disclosure: Kaja Lukas: No conflict of interest disclosed. Anne Letsch: Financing of Scientific Research: BMS,Merck, Novartis; Expert Testimony: Celgene, Novartis

Introduction: Infectious complications are a major cause of mortality after allogeneic hematopoietic stem cell transplantation, which may largely reflect an impairment of immune memory function. Donor immune cells as part of the peripheral blood (PB) stem cell transplant contribute to an immediate protection. Subset distribution as well as homing potential and functionality of the transplant’s memory cells influence the outcome of the patient’s immune reconstitution. Therefore, we are analyzing the impact of G-CSF exposure on PB memory composition and functionality. Methods: We analyzed peripheral blood mononuclear cells (PBMCs) of adult healthy stem cell donors before and after four-day subcutaneous application of G-CSF in order to quantitatively, phenotypically and functionally analyze memory subsets via multicolor flow cytometry. In parallel, we studied bone marrow (BM) resident memory cells in femur heads from patients undergoing hip replacement surgery. Results: In our preliminary data, we could show that the absolute numbers of memory T and B cells in PB increased up to fivefold after treatment with G-CSF. We could find a higher proportion of CD69+ memory CD4 T cells (0.7 ± 0.3% vs. 2.2 ± 0.3%) and memory CD8 T cells (2.4 ± 0.8% vs. 6.9 ± 0.7%) after treatment with G-SCF. In contrast, CLA-expressing memory CD4 T cells (20.1 ± 2.9% vs. 11.2 ± 0.8%) and memory CD8 T cells (17.2 ± 4.3% vs. 10.1 ± 3.4%) declined. CD103 and CRTH2 expressing memory T cells increased, whereas the percentage of alpha4beta7 expression was similar before and after G-CSF. Compared to PB samples, BM memory cells exhibited a higher percentage of CD69 and CRTH2 expression, while CLA expression was negligible. Number of samples are still limited (n = 5) and significances are not achieved so far. Conclusion: Our paired analysis of donor PBMCs before and after mobilization identifies newly recruited memory subsets and potentially enables the delineation of their tissue origin. Based on this detailed characterization of the apheresis product in respect to the memory composition and functionality, we are currently studying the fate of the transferred memory cell subsets in the patients in order to analyze which human memory subsets and survival niches are able to promote long-term immune memory. Disclosure: No conflict of interest disclosed. P283

A prospective multicenter safety surveillance study with a 5% intravenous human immunoglobulin (IVIg) preparation, in patients suitable for IVIg treatment, with or without premedication Peinert S.1, Abenhardt W.2, Fenchel K.3, Hübner A.4, Reichert D.5, Scheidegger C.6, Schmidt A.7, von Wussow P.8, Galic M.9 Onkologie Westerstede Aurich Rhauderfehn, Rhauderfehn, Germany, Onkologie Elisenhof, Munich, Germany, 3MVZ Saaletal, Saalfeld/Saale, Germany, 4Schwerpunktpraxis für Hämatologie und Onkologie, Bayreuth, Germany, 5Onkologie Westerstede Aurich Rhauderfehn, Westerstede, Germany, 6 Nordbadpraxis, München, Germany, 7Onkologie Schmidt, Cottbus, Germany, 8 Facharzt für Innere Medizin, Hannover, Germany, 9Kedrion International, Wien, Austria 1 2

Introduction: We investigated the tolerability of a human immunoglobulin preparation (Ig VENA® 50g/l for infusion) in patients with primary immunodeficiencies or secondary immunodeficiencies e.g. after cytotoxic treatment for haemato-oncologic malignancies; and with autoimmune diseases like idiopathic thrombocytopenic purpura (ITP) and chronic

Oncol Res Treat 2016;39(suppl 3):1–104

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inflammatory demyelinating polyneuropathy (CIDP). The nature and frequency of adverse drug reactions (ADR) were recorded under routine clinical conditions or with premedication administered prior to infusions. The study was conducted over a four year period. Methods: A multicenter, prospective, non-interventional study conducted in Germany. Subjects received 5% IVIg according to the prescribing information. Dosage, tolerability, any premedication, and adverse events were recorded. Dose regimen was at the investigator´s discretion. Results: 202 patients (120 f, 82 m) received a total of 1,218 infusions. IVIg was used as replacement therapy in immunodeficiency syndromes in 92.6% of patients (n = 187/202), and for immunomodulation in 7.4% of patients (n = 15/202), with a mean total dose per infusion of 15.9 g and 31.2 g, respectively. Across all indications the tolerability of IVIg infusions was judged by the physicians as very good or good in 99.2% (n = 1,208/1,218; very good n = 972/1,218, good n = 236/1,218) of the infusions even in the absence of premedication in 92% of infusions. Where premedication was given, mainly antihistamines and steroids were used. In total 8 non-serious ADRs were reported in 7 patients. None of the patients who had received premedication had an ADR; there were no serious ADRs reported during the study period. Conclusions: Treatment with 5% IVIg demonstrated an excellent tolerability profile when used under routine clinical conditions and in the absence of premedication, in the vast majority of infusions. Disclosure: Stefan Peinert: Financing of Scientific Research: Honorarium received from Kedrion international; Expert Testimony: Kedrion Internaional was the study sponsor Maja Galic: Employment or Leadership Position: Medical Director at Kedrion International; Expert Testimony: Kedrion Internaional was the study sponsor P284

Programmed death ligand 1 serum levels may reflect immunosuppression in patients with advanced renal cell cancer under treatment with a gene-modified allogeneic tumor cell vaccine (RCC26 (IL-7/CD80)) Flörcken A.1,2, Panzer I.1,2, Kopp J.3, Dörken B.1,2, Blankenstein T.4,5, Pezzutto A.6, Westermann J.1,2 Charité University Medicine, Campus Virchow-Klinikum, Department of Hematology, Oncology, and Tumor Immunology, Berlin, Germany, 2Labor Berlin Charite´ Vivantes GmbH, Berlin, Germany, 3Charité University Medicine Berlin, Campus Berlin-Buch, Experimental and Clinical Research Center, Berlin, Germany, 4Max Delbrück Center for Molecular Medicine, Berlin, Germany, 5 Charité University Medicine Berlin, Campus Benjamin Franklin, Institute of Immunology, Berlin, Germany, 6Charité University Medicine Berlin, Campus Benjamin Franklin, Department of Hematology, Oncology, and Tumor Immunology, Berlin, Germany 1

Introduction: Tumor-induced immunosuppression is a hallmark of cancer and a major challenge for immunotherapy. We have previously reported on a clinical phase-I trial with an allogeneic, HLA-A2-matched RCC cell line transfected with IL-7 and CD80 as a vaccine in RCC patients (Westermann et al. 2011), showing severe immunosuppression at baseline. Interestingly, an association with the nuclear factor-kappa B (NF-kB) signaling pathway could be established, implying that suppression of NFkB may play a key role for tumor-induced immunosuppression in RCC patients (Flörcken et al. 2015). Methods: To further understand why the allogeneic gene-modified (IL-7/ CD80 co-transfected) renal cell cancer vaccine failed to induce clinically relevant TH-1-polarized immune responses, PD-L1 serum levels were analyzed both in patients that had been treated within the vaccination study (n = 6) and in healthy donors (n = 10). Additionally, PD-L1 serum levels were correlated with clinical and immunological data from our previous studies. Results: Before vaccination, highly elevated levels of soluble PD-L1 were measured in RCC patients (mean sPD-L1 in ng/ml =1.68) as compared to healthy controls (mean sPD-L1 in ng/ml = 0.56) (p < 0.001). Interestingly, the vaccine was able to reduce soluble PD-L1 levels in RCC patients (mean sPD-L1 before vaccination:1.68 ng/ml, after vaccination: 1.35 ng/ml, n.s.),

Abstracts

however normal soluble PD-L1 serum levels were not achieved. Furthermore, reduction of PD-L1 serum levels was associated with a longer PFS, although this correlation did not reach statistical significance. Conclusion: To the best of our knowledge, this is the first report on soluble PD-L1 serum levels during treatment with a gene-modified cancer vaccine. Our data suggest that measurement of PD-L1 in cancer patients might be a useful tool for both the quantification of immunosuppression and the monitoring of immune responses within immunotherapy trials. Disclosure: Anne Flörcken: Financing of Scientific Research: Bayer Healthcare, Pfizer Pharma; Other Financial Relationships: Pfizer Pharma, Bayer Healthcare Jörg Westermann: No conflict of interest disclosed. P285

The deubiquitinase inhibitor b-AP15 and its effect on phenotype and function of monocyte-derived dendritic cells Altdoerfer V.1, Kropp K.1, Haen S.P.1, Grünebach F.1, Rittig S.M.1, Kanz L.1, Salih H.R.1,2, Dörfel D.1,2 Eberhard Karls University, Department of Hematology and Oncology, Tübingen, Germany, 2Clinical Collaboration Unit Translational Immunology, German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Partner site Tübingen, Tübingen, Germany 1

Introduction: The therapeutic efficacy of proteasome inhibition is well established in multiple myeloma and mantle cell lymphoma, which has led to its approval and incorporation in treatment algorithms for these malignancies. The first-in-class drug bortezomib impairs proteasome function by inhibition of the 20S core particle. However, proteasome function can also be targeted by inhibition of the 19S deubiquitinase (DUB) activity. Recently, b-AP15 was identified as a novel small molecule DUB inhibitor that inhibits growth of cancer cells in preclinical analyses (D´Arcy et al., Nat. Med. 2011). As bortezomib impairs several immune properties of monocyte-derived dendritic cells (DCs), in the current study we analyzed the influence of b-AP15 on DC phenotype and function. Methods: DCs were generated from PBMC of healthy donors. Plastic adherent monocytes were cultured in RP10 medium supplemented with granulocyte macrophage-colony-stimulating factor (100 ng/mL) and interleukin-4 (20 ng/mL). The medium was replenished with cytokines every other day and different concentrations of b-AP15 (10 nM, 100 nM, 500 nM, 1000 nM) were added to the culture on day 6. IL-10- (10 ng/mL) and bortezomib-treated (10 ng/mL) DCs served as controls. For maturation, lipopolysaccharide (LPS; 100 ng/mL) was added on day 6 and DCs were harvested on day 7 for immunophenotyping and functional analyses. Results: Immunophenotyping on day 7 revealed a clear downregulation of CD1a, CD83, CD86 and CD80 by bortezomib and IL-10 in mature DCs compared to untreated controls. In contrast, no influence of b-AP15 on these immunoregulatory molecules and maturation markers was observed upon exposure to b-AP15 compared to the untreated control. In line with differential effects of the two proteasome inhibitors on the immunophenotype, b-AP15 did also not impair DC migration and stimulatory capacity, while comparable effects of both compounds on apoptosis, susceptibility to NK cell killing and metabolic acticitiy were observed with various mantle cell lymphoma cell lines. Conclusion: In summary, our results suggest that b-AP15 mediates similar antitumor efficacy as bortezomib while preserving the immunostimulatory capacity of DCs, indicating that b-AP15 may be exquisitely suitable for combinatory treatment approaches. Disclosure: No conflict of interest disclosed.

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Posterdiskussion Lungentumoren P286

Combining anti-VEGF therapy and immune checkpoint inhibitors improves outcome in small cell lung carcinomas Meder L.1,2, Schuldt P.2, Vlasic I.1,3, Volz C.1,2, Golfmann K.1,2, Zaplatina A.1,2, Florin A.2,4, Tharun L.2,4, Büttner R.2,4, Reinhardt C.1,2,3, Ullrich R.1,2 University Hospital Cologne, Department of Internal Medicine I, Cologne, Germany, 2University Hospital Cologne, Lung Cancer Group Cologne, LCGC, Cologne, Germany, 3University of Cologne, Cluster of Excellence in Cellular Stress Responses in Aging-associated Diseases, CECAD, Cologne, Germany, 4 University Hospital Cologne, Institute of Pathology, Cologne, Germany 1

Metastatic lung cancer is the leading cause of smoking- and cancer-related deaths worldwide. SCLC is the most aggressive lung cancer and is driven by lesions in RB1 and TP53. SCLC accounts for approximately 14% of lung cancers with a median survival of 9 months, diagnosed at late stage. Thus, SCLC is usually not treated by surgery but with radiation- or chemotherapy. Results from KEYNOTE-028 were presented on the ASCO conference 2015. Here, programmed death ligand 1 (PD-L1) positive late stage SCLC patients received high-affinity, humanized monoclonal programmed death receptor 1 (PD-1) antibody pembrolizumab (MK-3475). They showed response rates of up to 25% which is remarkable for SCLC, since targeted therapy for this tumor entity are rare. However, most SCLC patients harbor a primary resistance or acquire resistance during treatment by an immune suppressive microenvironment or other resistance mechanisms triggered by the tumor cells themselves. Thus, there is a critical need to combine immune checkpoint inhibitors with other therapies to overcome these resistances. We implemented a combined therapy concept including anti-VEGF and anti-PD-L1 monoclonal antibody therapy for SCLC in mouse models. As a read out we used X-ray computed tomography, flow cytometry and end point immunohistochemistry. We found significantly increased progression-free survival (PFS) upon anti-VEGF and anti-PD-L1 mono-therapy, which was prolonged upon combined therapy. Moreover, we identified in murine SCLC tumors which acquired resistance upon anti-PD-L1 mono-therapy, a significant increase in PD1+/TIM3+ cytotoxic T cells. This exhausted T cell phenotype was significantly reverted upon combined anti-VEGF and anti-PD-L1 therapy. Furthermore, anti-VEGF mono-therapy provided an increased metastatic potential to SCLC cells which was significantly reduced upon combination with anti-PD-L1 therapy. Taken together, there is evidence for the benefit of implementing combined anti-angiogenic and anti-immune checkpoint therapy approaches in the clinic in order to overcome acquired resistances. However, it remains a compulsive issue in SCLC to improve not only the PFS but also the the overall survival and to prolong the duration of therapy responses. Disclosure: No conflict of interest disclosed.

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MYSTIC – A Phase III, randomized, open-label study of Durvalumab (MEDI4736) in combination with Tremelimumab or Durvalumab alone versus platinum-based chemotherapy in first-line treatment of patients with advanced stage IV non small cell lung cancer (aNSCLC): A German update von Pawel J.1, Fischer J.2, Alt J.3, Rittmeyer A.4, Wehler T.5, Laack E.6, Griesinger F.7, Schneider C.P.8, Panse J.9, Dieing A.10, Rawluk J.11, Serke M.12, Kropf-Sanchen C.13, Lerchenmüller C.14, Bohnet S.15, Bischoff H.16, Nusch A.17, de Wit M.18, Rupprecht M.19, Alt A.20, McIntosh S.21, Rizvi N.A.22, Seggewiß-Bernhardt R.23 Asklepios Klinik Gauting GmbH – Betriebsstätte Gauting, Gauting, Germany, 2Lungenkrebszentrum Klinik Löwenstein, Löwenstein, Germany, 3 Universitätsmedizin der Johannes Gutenberg-Universität Mainz, Medizinische Klinik III, Mainz, Germany, 4Fachklinik für Lungenerkrankungen Immenhausen, Immenhausen, Germany, 5Universitätsklinikum des Saarlandes, Homburg/Saar, Germany, 6Hämato-Onkologie Hamburg, Hamburg, Germany, 7Pius-Hospital Oldenburg, Oldenburg, Germany, 8Zentralklinik Bad Berka GmbH, Bad Berka, Germany, 9Uniklinik RWTH Aachen, Aachen, Germany, 10Vivantes Klinikum Am Urban, Berlin, Germany, 11Universitätsklinikum Freiburg, Freiburg, Germany, 12 Lungenklinik Hemer, Hemer, Germany, 13Universitätsklinikum Ulm, Ulm, Germany, 14Praxis GEHO Münster, Münster, Germany, 15Universitätsklinikum Schleswig-Holstein, Lübeck, Germany, 16Thoraxklinik Heidelberg gGmbH, Heidelberg, Germany, 17Praxis für Hämatologie und internistische Onkologie, Velbert, Germany, 18Vivantes Klinikum Neukölln, Berlin, Germany, 19 AstraZeneca GmbH, Medical Affairs, Wedel, Germany, 20AstraZeneca GmbH, Clinical Operations, Wedel, Germany, 21AstraZeneca Ltd., Global Medicines Development, Macclesfield, United Kingdom, 22Columbia University Hospital, New York, United States, 23Universitätsklinikum Würzburg, Würzburg, Germany 1

Introduction: Standard of care (SoC) therapy in 1st-line NSCLC is a platinum-based regimen for EGFR- and ALK-Wildtype-patients (pts). However, durable benefit is rare owing to most developing resistance to chemotherapy. Currently, immune checkpoint inhibitors are a highly promising new approach. Preclinical data suggest that blocking both, the inhibitory CTLA-4 as well as PD-1 receptors or the ligand, PD-L1, is non-redundant and has a synergistic effect to restore the immune-mediated antitumor response through T-cells. Durvalumab is a selective, high affinity human IgG1 monoclonal antibody (mAb) that blocks PD-L1 binding to PD-1 and CD80, and Tremelimumab is a selective human IgG2 mAb inhibitor of CTLA-4. Durvalumab monotherapy has already shown durable responses in NSCLC and Durvalumab plus Tremelimumab has shown encouraging clinical activity and a manageable safety profile in pts with NSCLC. A comprehensive clinical development program of Durvalumab +/- Tremelimumab in NSCLC is underway. MYSTIC (NCT02453282) is a randomized, open-label, multicenter, global, Phase III study to determine the efficacy and safety of Durvalumab plus Tremelimumab or Durvalumab monotherapy versus SoC platinum-based doublets in the 1st-line treatment of pts with NSCLC. In Germany, 21 sites are participating in the trial. Methods: In MYSTIC, immunotherapy- and chemotherapy-naïve pts with NSCLC who are wild-type for EGFR and ALK will be randomized (1:1:1) to receive Durvalumab (20 mg/kg IV every 4 weeks for up to 12 months) plus Tremelimumab (1 mg/kg IV every 4 weeks for up to 4 doses); Durvalumab monotherapy (20 mg/kg IV every 4 weeks for up to 12 months); or SoC chemotherapy. Stratification factors are PD-L1 status and histology. The primary endpoint is progression-free survival (PFS). Secondary endpoints will further assess objective response rate, duration of response, proportion of pts alive and progression free at 12 months, time from randomization to second progression, and overall survival; safety (CTCAE v4.03) and tolerability; health-related quality of life; pharmacokinetics; and immunogenicity. Exploratory outcomes include analysis of potential biomarkers of response to treatment, a comprehensive analysis of the tumor immune activation landscape by IHC, as well as tumor mutation load, are assessed on a biopsy at trial entry. Recruitment is ongoing. To date, 95 pts out of 675 have been randomized in Germany since First Subject In was on Oct 2015. Disclosure: No conflict of interest disclosed.

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Anti angiogenic treatment induces tumor cell invasion and metastasis via EphA2 signaling in NSCLC Volz C.1, Zaplatina A.1, Siobal M.1, Chatterjee S.1, Schöttle J.1, Meder L.1, Florin A.2, Koker M.1, Buettner R.2, Miao H.3,4, Wang B.3,4, Hallek M.1, Acker-Palmer A.5, Heukamp L.C.2, Thomas R.K.2,6, Ullrich R.1 University Hospital Cologne, Department of Internal Medicine I, Center for Molecular Medicine and Center for Integrated Oncology, Cologne, Germany, 2 University Hospital Medical School, Institute of Pathology, Cologne, Germany, 3 Rammelkamp Center for Research, MetroHealth Medical Center, Case Western Reserve University School of Medicine, Cleveland, United States, 4Case Western Reserve University School of Medicine, Department of Pharmacology and Oncology, Cleveland, United States, 5University of Frankfurt, Institute for Cell Biology and Neuroscience, Frankfurt, Germany, 6University of Cologne, Medical Faculty, Department of Translational Genomics, Cologne, Germany 1

VEGF / VEGFR-2 signaling is one of the main driving forces during tumor angiogenesis. However, clinical findings have shown that VEGF / VEGFR-2 targeted therapies provide only little efficacy in tumor shrinkage or benefit in survival. Moreover, recent preclinical studies reported that VEGF targeted therapies conditioned the tumor cells to aggressive invasiveness and metastasis. Using a spheroid invasion model we found that drug-induced inhibition of VEGFR2 on tumor cells drives tumor cell migration. Biochemically, we found that VEGFR2 inhibition on tumor cells induces serine phosphorylation of EphA2. Drug-induced inhibition of tumor VEGFR2 releases a newly identified EphA2/VEGFR2 heterocomplex, thereby allowing AKT to phosphorylate Serine 897 of EphA2. This VEGFR2 inhibition induced EphA2-Serine 897-phosphorylation mediates an EMT-like transition. Selective genetic modeling of the Serine 897 of EphA2 or drug-induced inhibition of EphA2 abrogates the formation of metastases in vivo upon VEGFR2 inhibition. In summary, our findings indicate that VEGFR2 targeted therapy conditions VEGFR2-positive NSCLC to Serine 897 EphA2-dependent aggressive tumor growth and metastasis. These data shed light on the molecular mechanisms explaining the limited efficacy of VEGFR2 targeted anti-angiogenic treatment in lung cancer patients. Disclosure: No conflict of interest disclosed. P289

Results from a multi-center, open-label, phase II study investigating the combination of RAD001 (everolimus) with paclitaxel and carboplatin in first line treatment of patients with advanced (stage IV) large cell lung cancer with neuroendocrine differentiation (LCNEC) Engel-Riedel W.1, Grohé C.2, Kropf-Sanchen C.3, von Pawel J.4, Gütz S.5, Kollmeier J.6, Eberhardt W.7, Christopoulos P.8, Nimmrich I.9, Sieder C.9, Baum V.9, Serke M.10, Thomas M.8 Kliniken der Stadt Köln, Lungenklinik, Köln, Germany, 2Evangelische Lungenklinik Berlin, Berlin, Germany, 3Universitätsklinikum Ulm, Ulm, Germany, 4 Asklepios Fachkliniken München-Gauting, München-Gauting, Germany, 5 Evangelisches Diakonissenkrankenhaus Leipzig, Leipzig, Germany, 6Helios Kliniken Berlin, Berlin, Germany, 7Universitätsklinikum Essen, Essen, Germany, 8 Thoraxklinik Heidelberg, Heidelberg, Germany, 9Novartis Pharma GmbH, Nürnberg, Germany, 10Lungenklinik Hemer, Hemer, Germany 1

Introduction: Large cell neuroendocrine carcinoma of the lung (LCNEC) make up approximately 3% of all lung cancers. These tumors in general have a bad prognosis and currently there are only very limited treatment options, including platinum derivatives and etoposide.The PI3/AKT/ mTOR pathway is known to be dysregulated in neuroendocrine tumors (NETs). As the mTOR inhibitor RAD001 (everolimus) already has proven effectiveness in different types of NETs including carcinoids and pancreatic NETs, we tested whether RAD001 might be also an effective treatment option in advanced LCNEC patients Methods: In the presented Phase II trial, daily RAD001 was combined with carboplatin and paclitaxel in patients with histologically confirmed stage IV LCNEC according to WHO criteria. Further inclusion criteria

Abstracts

were measurable disease according to RECIST 1.1 and adequate bone marrow, renal, and liver function. Main exclusion criteria were symptomatic CNS metastases and prior treatment for advanced LCNEC. Enrolled patients received daily everolimus in combination with 4 cycles of carboplatin and paclitaxel, followed by RAD001 maintenance therapy. The primary objective was to evaluate the efficacy of this treatment by assessing the proportion of progression-free patients after three months of treatment. Results: Ten German trial sites enrolled altogether 49 patients to the trial (mean age: 62 ± 9 years; 71% men). The primary endpoint (progression-free at month 3) was achieved by 24 patients (49%), assessed by an independent central imaging reviewer. Further endpoints were an overall response rate (ORR) until month 3 of 45%, a disease control rate (DCR) until month 3 of 73.5%, a median progression-free survival (PFS) of 4.3 months, and a median overall survival (OS) of 9.8 months. At least one toxicity occurred in 86% of all enrolled patients with grade 3/4 toxicities in 51%. Most frequent toxicities were diarrhea, fatigue, anemia, and neutropenia. Conclusions: The presented results show that a combined therapy of carboplatin and paclitaxel with the mTOR inhibitor RAD001 is an alternative treatment option for LCNEC patients. When comparing to other trials, the effectiveness is comparable to a treatment regimen of cisplatin and etoposide. Disclosure: Walburga Engel-Riedel: Advisory Role: Advisory Boards; Financing of Scientific Research: Lilly, Boehringer-Ingelheim, Novartis, Roche Michael Thomas: Financing of Scientific Research: Astrazeneca, Roche, Novartis, Pfizer, MSD, BMS, Lilly, Boehringer P290

Economic burden of clinical trials in lung cancer in a German Comprehensive Cancer Center Kron F.1, Kostenko A.1, Scheffler M.1, Glossmann J.1, Fischer R.1, Michels S.1, Nogova L.1, Hallek M.1, Zander T.1, Wolf J.1, Lung Cancer Group Cologne Uniklinik Köln, Klinik I für Innere Medizin, Köln, Germany

Introduction: Recent success of individualized lung cancer therapy has triggered fundamental changes in clinical research strategies. To date there is a strong focus on early proof of concept trials in genetically preselected, often rare, patient subgroups. From a hospital management perspective, this analysis focuses on the economic burden caused by such trials for advanced lung cancer patients in a German Comprehensive Cancer Center (CCC). Methods: Clinical and economic data from trials for advanced lung cancer (LC), pharma-sponsored trials (PhST) as well as investigator initiated trials (IIT), conducted between 2011 and 2015 at the Center for Integrated Oncology (CIO) Cologne, were analyzed using a profit-center calculation model. Results: 161 patients were enrolled in 27 clinical trials including 2,173 trial visits in total (PhST: 1,319, IIT: 854). We have identified `trial visit` as a key economic parameter determining costs and payments. In comparison of two groups (A≤3; B>3 patients enrolled) we found negative profit margins of €-1,444 per patient (PP) in A and positive in B (€217). Significant differences were also found between PhST and IIT (p = 0.009) concerning the number of visits PP, not, however, with regard to duration of treatment PP. Additionally, sub analysis show structural differences in cost composition by conducting PhST and IIT. Conclusion: Individualized, genomically targeted LC therapies goes along with an increase of early phase trials in rare genetic subgroups and, consequently, low patient recruitments. For trials with low patient numbers and IIT, it is difficult to break even with the costs. These observations demonstrate the need for CCC to thoroughly recalculating the costs of such trials in order to receive adequate, cost-covering compensation by pharmaceutical companies. They also underline the need for novel structures in public funding for investigator-initiated clinical trials in precision medicine. Disclosure: No conflict of interest disclosed.

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P292

Detection of an EGFR kinase domain duplication in a lung adenocarcinoma patient by liquid biopsy using hybrid capture based next generation sequencing

Implementation of the lung-cancer biology and outcome (LuCa-BiO) study integrating routine clinical data, quality of life assessment and liquid biopsy biobanking

Wiest G.1, Kohlhäufel M.2, Müller J.3, Lakis S.3, Wesseler C.1, Mariotti E.3, Zacherle T.3, Leenders F.3, Gloeckner C.3, Heuckmann J.M.3, Menon R.3, Heukamp L.3

Schlenk R.F.1, Schmidtke-Schrezenmeier G.2, Neagoie A.M.1, Schmelzle B.1, Nagel G.3, Buske C.4, Wessendorf S.5, Schwänen C.5, Hetzel M.6, Liewald F.7, Brinkmann F.8, Ott G.9, Sträter J.10, Hamel T.1, Kuhn P.11, Welke C.11, Lang G.12, Möller P.13, Marienfeld R.13, Döhner H.1, Krof-Sanchen C.2, Bullinger L.1

Asklepios Klinikum Harburg, Atemwegs-, Lungen- und Thoraxklinik, Hamburg, Germany, 2Klinik Schillerhöhe, Abteilung für Pneumologie und Thoraxonkologie, Gerlingen, Germany, 3NEO New Oncology, Cologne, Germany 1

Introduction: EGFR kinase domain duplications are a rare but targetable alteration, driving lung cancer. Only limited patient data has been published on the detection of the kinase duplications. This molecular alteration has been shown to lead to the activation of EGFR and is thought to be predictive for response to treatment with EGFR inhibitors. Using a hybrid-capture based next-generation sequencing method, we describe here the initial detection of the EGFR kinase domain duplication in an FFPE sample from the primary tumor (NEOplus) and the subsequent detection in a liquid biopsy (NEOliquid) sample Method: A 72-year-old patient was diagnosed with a lung adenocarcinoma. Initial routine tests were negative for genomic alterations in known targetable lung cancer genes. Therefore, the patient was treated with 3 cycles of chemotherapy, but the patient continued to progress. Recently a tissue sample and a liquid biopsy was analyzed by NEO New Oncology (Cologne, Germany) using NEOplus and NEOliquid. The assay uses a hybrid-capture based next-generation sequencing assay that covers clinically relevant genomic alterations, such as point mutations, small insertions and deletions, selected gene fusions and copy number alterations within a panel of more than 90 genes for FFPE material or 30 genes for liquid biopsies. Results: Hybrid capture-based NGS assay were able to identify an EGFR kinase domain duplication. Based on the identification of the duplication, the patient was put on second line treatment with the EGFR inhibitor Afatinib. Despite a response of the primary tumor and the metastasis in lymph nodes and adrenal gland, a recent scan revealed that the metastasis in the brain and liver progressed. The discordance in response might depict the tumors heterogeneity or drug transportation and metabolism to the metastatic foci. Conclusion: Here we describe the detection of a rare EGFR kinase domain duplication using the NEO assay. Interestingly, when treated with Afatinib, some of the patient´s distal metastasis continued to progress. Novel technologies capable of detecting rare genomic alterations, in the routine setting, further stresses on the urgent need to develop and identify drugs to treat patients harboring these rare mutations. Disclosure: No conflict of interest disclosed.

Universitätsklinikum Ulm, Innere Medizin III, Ulm, Germany, Universitätsklinikum Ulm, Innere Medizin II, Ulm, Germany, 3Epidemiologie und Biometrie, Epidemiologie, Ulm, Germany, 4Universität Ulm, Experimentelle Tumorforschung, Ulm, Germany, 5Klinikum Esslingen, Innere Medizin, Esslingen, Germany, 6Krankenhaus vom Roten Kreuz Bad Cannstatt, Pneumologie, Internistische Intensivmedizin, Beatmungsmedizin und Allgemeine Innere Medizin, Stuttgart, Germany, 7Klinikum Esslingen, Esslingen, Germany, 8 Diakonie-Klinikum Stuttgart und OSP Stuttgart e.V., Stuttgart, Germany, 9 Robert-Bosch-Krankenhaus, Pathologie, Stuttgart, Germany, 10Institut für Pathologie Esslingen, Esslingen, Germany, 11Comprehensive Cancer Center Ulm, Ulm, Germany, 12Universitätsklinikum Ulm, Thoraxchirurgie, Ulm, Germany, 13 Universitätsklinikum Ulm, Pathologie, Ulm, Germany 1 2

Background: Despite several approaches in prevention and early detection of lung cancer the prognosis is still poor. Recently, emerging data on the molecular background of lung cancer introduced several new approaches in the systemic therapy especially in patients with advanced stages. The aim of the LuCa-BiO protocol (ClinicalTrials.gov: NCT02613637) is to set up a multicenter registry including clinical, molecular and quality of life data to allow implementation of focussed interventional clinical programs and healthcare research. Methods: We set up a multicenter registry with a validated and certified web-based registration platform separating personal patient-data for the trusted third party from pseudonymized data for the liquid-biopsy lab, the quality of life assessment center as well as for the clinical data center and protocol coordination. Sites receive at the same time an easy to handle registration documentation sheet containing all relevant information. Clinical data are extracted from cancer registry datasets required by law with regular updates. Material from diagnostic specimens is available at the participating centers. Quality of life (QoL) is assessed by the EORTC QLC-C30 and LC 13 questionnaires. Results: The trial started in May 2014 with an exploratory set-up phase with three clinical centers. Until May 2016, 400 patients were registered and for the analysis the first 242 patient with available cancer registry data were selected. Clinical data and liquid biopsies are available from all included patients, QoL data in 188 patients. Median age at registration was 67 years (range, 31-89 years), 62% of the patients were male, 82% were registered at initial diagnosis and 18% later on; 16% had small cell lung cancers (SCLC) and 84% Non-SCLC with 57% adenocarcinoma and 31% squamous-cell carcinoma. UICC staging at diagnosis was IA 18%, IB 14%, IIA 8%, IIB 4%, IIIA 14%, IIIB 12% and IV 29%. QoL data revealed at diagnosis a median global status of QoL 75% (range, 14-100%), a median function-related QoL of 76% (range, 7-100%) a median symptom-oriented QoL of 79% (range, 21-100%) and a median overall QoL of 58% (16100%). Conclusions: This first data from our LuCa-BiO study in lung cancer patients show the feasibility of a multicenter approach combining cancer registry data with QoL assessment and liquid biopsy biobanking. Molecular and treatment data are currently updated. Disclosure: No conflict of interest disclosed.

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Exploratory analysis of efficacy by histology and frontline therapies in a nonsquamous non-small cell lung cancer (NSCLC) subgroup in REVEL: A randomized phase III study of ramucirumab (RAM) plus docetaxel (DOC) vs DOC plus placebo (PBO) for second-line treatment of stage IV NSCLC

Expression of the orphan G-protein-coupled receptor GPR19 in different lung cancer entities

Schuette W.1, Reck M.2, Kimmich M.3, Schumann C.4, Paz-Ares L.5, Garon E.6, Pérol M.7, Zimmermann A.8, Lee P.9 Krankenhaus Martha-Maria Halle-Dölau, Klinik für Innere Medizin II, Halle (Saale), Germany, 2LungenClinic Grosshansdorf, Airway Research Center North (ARCN), Großhansdorf, Germany, 3Abt. Pneumologie und Pneumologische Onkologie Klinik Schillerhöhe, Zentrum für Pneumologie, Beatmungsmedizin und Thoraxchirurgie, Gerlingen, Germany, 4Klinik für Pneumologie, Thoraxonkologie, Schlaf- und Beatmungsmedizin, Kempten, Germany, 5 University Hospital Virgen del Rocío, Medical Oncology, Seville, Spain, 6David Geffen School of Medicine at UCLA, Translational Research in Oncology-US Network, Los Angeles, United States, 7Département de Cancérologie Médicale Centre Léon-Bérard, Lyon, France, 8Eli Lilly and Company, Indianapolis, United States, 9Eli Lilly and Company, Bridgewater, United States 1

Introduction: The phase III trial REVEL (NCT01168973) led to FDA and EMA approval of RAM + DOC for metastatic NSCLC patients with disease progression on/after standard platinum-based frontline chemotherapy (pbFCT). Pemetrexed (PEM) is standard of care in nonsquamous NSCLC within pbFCT. An exploratory efficacy analysis of RAM + DOC for nonsquamous NSCLC, including adenocarcinoma, is reported. Methods: Frontline therapy for nonsquamous patients was defined as PEM induction therapy ± any PEM maintenance therapy (PEM IT) and non-PEM induction therapy ± non-PEM maintenance therapy (non-PEM IT). Nonsquamous patients received DOC (75 mg/m2) + RAM (10 mg/kg; n = 465) or DOC + PBO (n = 447) after disease progression on pbFCT. Endpoints were overall survival (OS), progression-free survival (PFS) and objective response rate (ORR). OS and PFS were analyzed using the Kaplan-Meier method and the Cox proportional hazard model; ORR was analyzed using Cochran-Mantel-Haenszel test. Results: Of 912 patients with nonsquamous NSCLC (73%), the majority were adenocarcinoma (79%; N = 725). Baseline characteristics and poststudy therapy use in nonsquamous patients were balanced between PEM IT and non-PEM IT subgroups. Maintenance therapy use was higher for patients with prior PEM IT (37.9%) than non-PEM IT (13.7%). RAM + DOC efficacy for adenocarcinoma [OS = 11.2 months (HR, 95% CI: 0.83, 0.69−0.99); PFS = 4.5 months (HR, 95% CI: 0.75, 0.64−0.88); ORR = 19%] was consistent with nonsquamous [OS = 11.1 months (HR, 95% CI: 0.83, 0.71−0.97); PFS = 4.6 months (HR, 95% CI: 0.77, 0.67−0.88); ORR = 22%]. Results for RAM + DOC vs DOC + PBO were: PEM IT: OS = 11.8 months vs 9.0 months (HR, 95% CI: 0.779, 0.62−0.98); non-PEM IT: OS = 11.0 months vs 9.9 months (HR, 95% CI: 0.855, 0.68−1.07); PEM IT: PFS = 5.1 months vs 3.7 months (HR, 95% CI: 0.691, 0.56−0.85); nonPEM IT: PFS = 4.5 months vs 3.5 months (HR, 95% CI: 0.772, 0.63-0.94); PEM IT: ORR = 20.0% vs 14.9%; non-PEM IT: ORR: 24.0% vs 14.3%. Conclusions: In nonsquamous NSCLC, RAM + DOC improved OS, PFS and ORR regardless of prior PEM or non-PEM treatment. Favorable efficacy was seen for NSCLC histological subtypes, including adenocarcinoma. Disclosure: Wolfgang Schuette: Advisory Role: Yes; Financing of Scientific Research: Yes Pablo Lee: Employment or Leadership Position: Yes; Stock Ownership: Yes

Gerlach L.1, Kaemmerer D.2, Sänger J.3, Schulz S.1, Lupp A.1 Universitätsklinikum Jena, Institut für Pharmakologie und Toxikologie, Jena, Germany, 2Zentralklinik Bad Berka, Klinik für Allgemein- und Viszeralchirurgie, Bad Berka, Germany, 3Labor für Pathologie und Zytologie Bad Berka, Bad Berka, Germany 1

Introduction: Lung cancer is one of the leading causes of tumor-related death worldwide, not least since for most of the entities no efficient standard therapy is available so far. Therefore, the discovery of new markers to open up additional treatment options is urgently needed. Recently it has been shown that the mRNA of the orphan G-protein-coupled receptor (GPCR) GPR19 is highly expressed in small cell lung cancer (SCLC). Thus, the aim of the present study was to investigate the expression of this receptor at the protein level by means of immunohistochemistry not only in SCLC, but also in other lung cancer entities using a novel rabbit anti-GPR19 antibody. Methods: Overall, 320 samples from 132 lung cancer patients (typical carcinoid [TC]: n = 21; atypical carcinoid [AC]: n = 25; SCLC: n = 42; adenocarcinoma: n = 22; squamous cell carcinoma: n = 22), comprising both primary tumors and metastases, were evaluated for the GPR19 expression by means of immunohistochemistry. The immunohistochemical stainings were evaluated using the Immunoreactive Score (IRS), comprising values from 0 to 12 points, and correlated to clinical data. Tumors with IRS values ≥ 3 IRS points were considered positive. Results: With the only exception of the squamous cell carcinomas, the GPR19 could be detected at a high frequency in all lung cancer entities investigated. It was present in 95% of the TC, in 96% of the AC, and in 91% of the SCLC and of the adenocarcinomas. In contrast, only 23% of the squamous cell carcinomas were GPR19 positive. Also with respect to the intensity of expression, squamous cell carcinomas displayed distinctly lower IRS values as compared to the other tumor entities and there was also a significant difference between the values of the adenocarcinomas and those of the SCLC (median IRS values: TC: 8.0, AC: 7.6, SCLC: 9.0, adenocarcinomas: 6.1; squamous cell carcinomas: 1.5). Between primary tumors and metastases no significant difference in GPR19 expression was observed. Conclusions: Because of the high incidence and intensity of expression in TC, AC and SCLC, the GPR19 may serve as a good target for diagnostics and therapy especially of lung tumors with neuroendocrine background. Disclosure: No conflict of interest disclosed. P295

Clinical research platform into molecular testing, treatment and outcome of non-small cell lung carcinoma patients (CRISP): A prospective German registry in stage IV NSCLC – AIO-TRK-0315 Griesinger F.1, Eberhardt W.E.E.2,3, Marschner N.4, Jänicke M.5, Spring L.5, Sahlmann J.5, Karatas A.6, Hipper A.6, Sebastian M.7, Thomas M.8,9 Pius-Hospital Oldenburg, University of Oldenburg, Universitätsklinik Innere Medizin-Onkologie, Oldenburg, Germany, 2University Hospital Essen, Department of Medical Oncology, Essen, Germany, 3Ruhrlandklinik and University Duisburg-Essen, West German Cancer Centre, Essen, Germany, 4 Praxis für interdisziplinäre Onkologie und Hämatologie, Freiburg, Germany, 5 iOMEDICO, Freiburg, Germany, 6AIO-Studien-gGmbH, Berlin, Germany, 7 Klinikum der J.W. Goethe-Universität Frankfurt, Medizinische Klinik II Hämatologie/Onkologie, Rheumatologie, Infektiologie, HIV, Frankfurt am Main, Germany, 8Thoraxklinik am Universitätsklinikum Heidelberg, Heidelberg, Germany, 9German Center for Lung Research (DZL), Heidelberg, Germany 1

Background: Treatment in NSCLC is quickly evolving. Whether outcome and PRO data generated in clinical trials with narrow inclusion and exclusion criteria will hold up in the routine practice is of high interest, especially due to the increasing costs of new drugs. Therefore registry data are of increasing importance to patients, physicians and third parties. This is

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why we have started a prospective, clinical registry for patients with metastatic non-small cell lung cancer. Methods: The purpose of CRISP is to set up a national clinical research platform to document representative data on molecular testing, sequences of therapies and other treatment modalities, course of disease in patients with advanced or metastatic NSCLC not amenable to curative treatment. A particular focus is on molecular biomarker testing of patients before the start of first-line treatment. PRO assessment will provide large-scale data on quality of life and anxiety/depression for real-life patients in routine practice. In addition, two questionnaires (concerning individual quality of life and patient-caregiver communication) will be validated in German patients with metastatic NSCLC. Furthermore CRISP will set up a decentral tissue annotation for future collaborative, investigational scientific biomarker testing. This study will be carried out in up to 150 representative cancer centers in all therapeutic sectors in Germany. More than 5000 patients will be recruited and followed up to a maximum of 3 years, respectively until death. The first patients have been included as of December 2015. Results: Preliminary data will be presented at the meeting in terms of molecular test rates, demographic data as well as treatment stratification in the 1st line setting. Conclusion: The registry CRISP will be the first to present representative real life data, covering all treatment settings of patients with NSCLC in Germany. ClinicalTrials.gov Identifier: NCT02622581. CRISP is supported by Grants from AstraZeneca, Boehringer Ingelheim, Bristol-Myers Squibb, Celgene, Merck Sharp & Dohme, Novartis, and Pfizer. Disclosure: No conflict of interest disclosed. P296

Disclosure: Frank Griesinger: Advisory Role: Pfizer, Astra, Boehringer, Novartis, Roche, Celgene, MSD, BMS, Clovis; Financing of Scientific Research: Pfizer, Astra, Boehringer, Novartis, Roche, Celgene, MSD, BMS, Clovis; Expert Testimony: ASTRA Markus Tiemann: Advisory Role: Pfizer, Astra, Boehringer, Novartis, Roche, Celgene, MSD, BMS; Honoraria: Pfizer, Astra, Boehringer, Novartis, Roche, Celgene, MSD, BMS; Expert Testimony: Astra Zeneca P297

Non-Small Cell Lung Cancer (NSCLC) - treatment research and treatment reality in oncology practices (adjuvant therapy) Wilhelm S.1, Hutzschenreuter U.2, Innig G.3, Göttel R.4, Tessen H.-W.5, Projektgruppe Internistische Onkologie (PIO) Onkologische Schwerpunktpraxis, Güstrow, Germany, 2Onkologische Schwerpunktpraxis, Nordhorn, Germany, 3Onkologische Schwerpunktpraxis, Rheine, Germany, 4rgb Onkologisches Management GmbH, Sarstedt, Germany, 5 Onkologische Kooperation Harz, Goslar, Germany 1

Intercalated TKI and chemotherapy induction in EGFR mt+ NSCLC stages IIIA to IV OMD: report of 5 cases and phase II study Griesinger F.1, Lüers A.1, Roeper J.1, Falk M.2, Conradi I.S.1, Reinhardt M.3, Kluge A.4, Willborn K.5, Prenzel R.6, Scriba D.7, Henke R.-P.8, Hallas C.2, Netchaeva M.1, Tiemann M.2 Pius-Hospital Oldenburg, University of Oldenburg, Hämatolologie/Onkologie, Innere Medizin-Onkologie, Oldenburg, Germany, 2Hämatopathologie Hamburg, Pathologie, Hamburg, Germany, 3Pius-Hospital Oldenburg, Nuklearmedizin, Oldenburg, Germany, 4Pius-Hospital Oldenburg, Radiologie, Oldenburg, Germany, 5Pius-Hospital Oldenburg, University of Oldenburg, Strahlentherapie und Radioonkologie, Medizinische Physik, Oldenburg, Germany, 6Pius-Hospital Oldenburg, Innere Medizin, Pneumologie, Gastroenterologie, Oldenburg, Germany, 7Pius-Hospital Oldenburg, Thorax- und Gefäßchirurgie, Oldenburg, Germany, 8Pathologisches Institut Oldenburg, Pathologie, Oldenburg, Germany 1

Background: EGFR TKI treatment is standard of care in patients with stage IV NSCLC carrying an activating EGFR mutation. Induction concepts in locally advanced EGFR mt+ NSCLC including TKI have not been studied extensively. Recently new focus has been shed on intercalated regimens of chemotherapy and TKI, showing improved PFS as well as OS. This concept was used as induction regimen in 5 patients with activating EGFR mutation in stages IIIA and IIIB. Methods: Patients were diagnosed and worked up according to standard imaging, histology and immunohistology methods. EGFR, KRAS, BRAF, ALK and P53 mutation analysis were performed with standard procedures as described by Halbfass et al. 2013. Remission induction was measured by RECIST 1.1, regression grading by Junker criteria. Results: 3 f never or light smokers (pt #1, 3, 5), 59, 63, 62 y.o. and 2 m never or light smoker (pt# 2and 4) 57 and 70 y.o. had adenocarcinoma of the lung, 2 with L858R (#1,2)and 3 with E 19 Del (#3-5). 4/5 patients (#1-4) carried a a p53 mutation, case #1 a disruptive, cases #2-4 a non disruptive mutation. Stages were IIIB (#1-3), IV OMD with 2 metastases in 1 vertebra (#4) and IIIA (#5). Induction therapy was started with gefitinib 250 mg/die p.o. (#1,4,5) or erlotinib 150 mg/die p.o. days -12 to -1 (#3,4) in order to prove responsiveness of the tumour to EGFR-TKI. On day 0

partial response or no progression was achieved in all 5 pts. Therapy was continued with 3 cycles of Docetaxel 75 mg/m2 d1 and Cisplatin 50 mg/ m2 d 1 and 2 qd22 or Paclitaxel 200 mg/m2 + Carboplatin AUC 6.0 d1 in combination with Gefitinib 250 mg / die or Erlotinib 100 mg/ die d4-19. PR was was achieved after 2 cycles in all pts. All 5 pts were resected and regression grade IIB or III was observed in mediastinal lymph nodes and primary tumor in pts #1-4, only IIA and I in pt #5. 4/4 pts with prior N2 or N3 LN received postoperative radiotherapy. Patients #1, 3, 5 are in CR, patient 2 and 4 developed one isolated CNS metastasis which has been stereotactically irradiated. Patients’ survival times are 10+, 13+, 41+, 7+, 5+ months respectively. Conclusion: Intercalated TKI treatment is a promising treatment choice in patients with EGFR mt+ locally advanced NSCLC. A phase II trial (NeoIntercal) trial is currently under way in 9 German centers in stages II and III using Gefitinib in combination with induction taxane based chemotherapy, supported by AIO, ASTRA Zeneca and the University of Oldenburg.

Oncol Res Treat 2016;39(suppl 3):1–104

Introduction: According to S3 guidelines, patients with a NSCLC stage II or IIIA are to receive a cisplatin-based adjuvant combination chemotherapy for 4 cycles within 60 days after a R0 resection (recommendation level A). Stage IB requires an individual therapy decision. Patients with an adjuvant chemotherapy in phase-III trials lived a median of 66-94 months. Methods: Since 2003, 89 oncological practises from 16 federal states have been documenting 2,864 disease histories of patients with a non-small cell lung cancer within the framework of the Project team of Internal Oncology (PIO). 2,723 of the cases with a total of 6,135 therapies could be evaluated in the registry ONCOReg. 409 (15.0%) patients received an adjuvant chemotherapy, 305 (74.6%) intravenous vinorelbine/cisplatin. Detailed results are available for 291 patients from 42 practises. Results: Patients’ characteristics Gender: 209 (71.8%) male; 82 (28.2%) female Age at initial diagnosis (median): 63 (41-78) years; 8 (2.8%) > 75 years; 54 (18.6%) > 70 years Previous therapy: 7 (2.4%) patients UICC: 53 (18.2%) I; 126 (43.3%) II; 106 (36.4%) III; 6 n.s. (T1-2, Nx) Histology: 138 (47.4%) adenocarcinoma; 146 (50.2%) squamous cell carcinoma; 7 others Smokers/Non-smokers: 99 (34.0%)/107 (36.8%)/85 n. s. Therapy Period from surgery to start of therapy (median): 42 (15-191) days Number of cycles (median): 4 (1-6) End of therapy: 180 (61.9%) as scheduled; 42 (14.4%) toxicity; 24 (8.2%) refusal; 14 (4.8%) hospitalization; 31 others Radiotherapy: 62 (21.3%) The median monitoring period covers 19.4 (0.3-111.1) months. Distant metastases have been reported for 82 patients so far, a local recurrence for 34 patients and a recurrent neoplasm for 8 patients. 51 (62.8%) patients received a chemotherapy due to recurrences or distant metastases. The

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1st-line therapy for them consisted mainly in platinum derivatives (28 (54.9%), taxanes (17 (33.3%) und antimetabolites (15 (29.4%). Survival The median disease-free survival is at 32.2 months. UICC I: months not reached; II: 38.4 months; III: 23.0 months). The median overall survival has not been reached so far. The 3-year survival rate is at 70%. UICC I: not reached (3-year survival 81%); II: 91.6 months (70%); III: 56.2 months (65%). Smokers not reached (64%); non-smokers 75.1 months (71%) Conclusions: The adjuvant therapy of the NSCLC is performed in compliance to the approved guidelines in treatment research.The survival data reflect the existing data. Disclosure: Stefan Wilhelm: Employment or Leadership Position: Praxisinhaber- Gemeinschaftspraxis; Advisory Role: Lilly, Roche, Sozialgutachten, Gerichte; Financing of Scientific Research: Amgen, Medac, Lilly, Mundipharm, Roche, Boehringer Ingelheim, GlaxoSmithKline, Novartis, AstraZeneca Hans-Werner Tessen: No conflict of interest disclosed. P298

Successful intercalating multimodal treatment strategy in a patient with advanced adenocarcinoma of the lung harboring the uncommon complex EGFR mutation L833V/H835L Frille A.1, Sändig I.2, Wirtz H.1 Universitätsklinikum Leipzig, Pneumologie, Leipzig, Germany, Universitätsklinikum Leipzig, Institut für Pathologie, Leipzig, Germany

1 2

Introduction: According to international guidelines for the treatment of advanced non-small cell lung cancer (NSCLC), sensitizing mutations of the epidermal growth factor receptor (EGFR) are predictive for response to the EGFR tyrosine kinase inhibitors (TKI) gefitinib erlotinib and afatinib in terms of improved response rate, progression-free survival, quality of life, and even overall survival. However, efficacy of those TKI for uncommon mutations in the EGFR gene may be distinct from common mutations like exon 19 deletions and L858R. Case: A 63-year-old male Caucasian never-smoker in a good performance status was diagnosed with a poorly differentiated adenocarcinoma of the right lower lobe of the lung. Fluorodeoxyglucose positron emission tomography (FDG-PET) revealed in addition hypermetabolic subcarinal, paratracheal, and supraclavicular lymph nodes on the right side being suspicious for malignancy. The initial clinical TNM classification was cT2a cN3 cM0 (stage IIIB). A complex mutation of L833V and H835L in exon 21 of the EGFR gene was detected from the tumor specimen by nucleic acid amplification via polymerase chain reaction followed by Sanger sequencing of EGFR from exon 18 to exon 21. Common drug-sensitive mutations including exon 19 deletions and L858R were not found. An oral regimen of the TKI gefitinib 250 mg per day was started. Since eight weeks of TKI treatment showed a stable disease, we decided to perform an intercalating chemotherapy containing cisplatin/pemetrexed over four courses leading now to a partial response of the pulmonary target lesion. Chemotherapy was then switched to second generation TKI afatinib 40 mg per day for six weeks resulting in a further partial response. After multidisciplinary discussion, the patient then underwent curative surgery with resection of right lower lobe and systemic lymphadenectomy revealing a postoperative ypT1b pN2 cM0, L1, V0 Pn0, R2 (supraclavicular lymph nodes). After recovery form surgery, an adjuvant concurrent chemoradiotherapy of the supraclavicular and mediastinal lymph nodes was performed and one course of cisplatin/pemetrexed was administered. Last restaging showed no evidence of suspicious hypermetabolic lesions in FDG-PET. Conclusion: Evidence predicting the efficacy of TKI for uncommon complex mutations of the EGFR is often lacking. Patients with a very good performance status despite an advanced N3-disease may need an intense discussion about individual multimodal treatment plan. Disclosure: No conflict of interest disclosed.

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Adenocarcinoma in pleural tissue in a patient after double lung transplantation: the question of tumor origin and therapeutic considerations Ufen M.-P.1, Janssen C.1, Müller L.1 Onkologie Unter Ems, Leer, Germany

Introduction: Lung transplantation may help to improve the prognosis of patients with end-stage lung diseases. The pre-interventional screening of donor and recipient organs is responsible for the successful outcome of lung transplantation, for example by detection of a malignant tumor in patients with severe pulmonary comorbidities. Nevertheless, the occurrence of a lung cancer after lug transplantation is rarely reported in literature. Case report: We report on the case of a 61 years old male patient with an double lung transplantation due to an combined pulmonary fibrosis and emphysema (CPFE) with a history of heavy smoking (50 py). Immunosuppression was performed with everolimus, MMF and steroids. A decortication of the left lower lobe were carried out 11 months post transplantation due to a persistent pleural effusion. The histological examination of pleural biopsies showed a low differentiated adenocarcinoma (K-RAS-mutated), classified as cTx cNx pM1a (PLE). Pathologic workup failed to detect a definite allocation of primary tumor. Staging diagnostics by PET-CT and intestinoscopy provided no evidence for an other primary. Consequently a pulmonary origin must be supposed, although no tumor was found in the donor lung. A pathologic examination of the explanted lung has not been performed. The pleural carcinomatosis defined an advanced disease, therefore a systemic treatment in palliative intention was carried out with pemetrexed only because of a reduced physical condition based on a pulmonary embolism. The first restaging after two months of treatment showed no evidence of disease except a small pleural effusion, which cannot be examined by taking a sample. Conclusions: Although the primary tumor cannot be determined without any doubt-a origin in the donor or recipient lung must be supposed. In accordance with the transplant centre, a malignancy of the explanted lung seems to be more likely due to the smoking history. In future we would recommend further pathological examination of the explanted organ to solve the question of origin. Remarkably, there is no definitive tumor manifestation detectable under the therapy with pemetrexed and everolimus. The addition of a m-TOR Inhibitor may have influenced the treatment with pemetrexed and represents potentially a novel therapeutic concept in lung cancer. Disclosure: No conflict of interest disclosed. P300

Bioluma: Biomarkers for nivolumab and evaluation of nivolumab plus ipilimumab in lung cancer – A phase II trial of nivolumab in combination with ipilimumab to evaluate efficacy and safety in lung cancer and to evaluate biomarkers predictive for response to immune checkpoint inhibition Fischer R.N.1, Abdulla D.1, Michels S.1, Nogova L.1, Brandes V.1, Scheffler M.1, Schäfer S.2, Scheel A.2, Thurat M.1, Vehreschild M.3, Thomas R.4, von Bergwelt-Baildon M.5, Büttner R.2, Wolf J.1 Uniklinik Köln, Klinik I für Innere Medizin, Lunge Cancer Group Cologne, Köln, Germany, 2Uniklinik Köln, Institut für Pathologie, Köln, Germany, 3Uniklinik Köln, Klinik I für Innere Medizin, German Centre for Infection Research, Köln, Germany, 4Universität Köln, Institut für Translationale Genomik, Köln, Germany, 5 Uniklinik Köln, Klinik I für Innere Medizin, Interventionelle Immunologie, Köln, Germany 1

Introduction: Checkpoint inhibition is a promising strategy in the treatment of various cancer entities. The anti-PD-1 antibody nivolumab recently was approved for secondline treatment in squamous and non-squamous non-small cell lung cancer (NSCLC). In small cell lung cancer (SCLC), preliminary data from ongoing clinical trials show promising

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results even in platinum-refractory subjects. However, the achievement of response rates of approximately 20% implicates the unmet need to further enhance therapeutic activity of checkpoint inhibition. In addition, strategies to better define patients responding to checkpoint inhibition before therapy, i.e. to identify predictive biomarkers need to be developed. Here, we present the ongoing clinical trial BIOLUMA which aims to evaluate efficacy and safety of adding ipilimumab to nivolumab in the treatment of nivolumab-refractory NSCLC patients and to evaluate efficacy and safety of upfront combination therapy of nivolumab and ipilimumab followed by nivolumab monotherapy in subjects with SCLC. In addition, a broad biomarker program is included in order to identify biomarker predictive for response and resistance to checkpoint inhibition. Methods: BIOLUMA is a multicentric non-randomised phase II trial in patients with non-squamous NSCLC and patients with SCLC after failure of platinum-based first-line therapy. In 12 German trial centres, NSCLC patients are treated with nivolumab until disease progression and subsequently receive a combination therapy of nivolumab and ipilimumab. SCLC patients receive four cycles of nivolumab in combination with ipilimumab and subsequent nivolumab monotherapy. Primary endpoint for the NSCLC cohort is ORR after addition of ipilimumab to nivolumab treatment. For the SCLC cohort, primary endpoint is ORR of the upfront combination therapy nivolumab and ipilimumab. In addition, extensive analysis of tumor biopsy material, peripheral blood and gut microbiome is performed. Tumor cells and tumor microenvironment are characterized by histology and immunohistochemistry. The role of specific somatic mutations and overall mutational load is analyzed by whole genome or whole exome sequencing (WGS, WES), neoepitope prediction and modeling of HLA-processing. Cellular and soluble blood components are assessed by FACS and ELISA-assays. Intestinal microbiome samples are analyzed by deep-sequencing. Disclosure: Rieke Fischer: Advisory Role: Teilnahme an Advisory Boards Bristol-Myers Squibb; Financing of Scientific Research: Honorare von Bristol-Myers Squibb für Vorträge, Teilnahme an AD Jürgen Wolf: Advisory Role: Teilnahme an Advisory Boards Bristol-Myers Squibb; Financing of Scientific Research: Honorare von Bristol-Myers Squibb für Vorträge, Teilnahme an AD

Posterdiskussion

Kolorektale Karzinome P301

SATB1 as a therapeutic target candidate in colorectal cancer: preclinical studies Frömberg A.1, Rabe M.1, Linnebacher M.2, Aigner A.1 Universität Leipzig, Medizinische Fakultät, Rudolf-Boehm-Institut für Pharmakologie und Toxikologie, Selbständige Abteilung Klinische Pharmakologie, Leipzig, Germany, 2University of Rostock, Department of General, Thoracic, Vascular and Transplantation Surgery, Rostock, Germany 1

Introduction: The Special AT-rich Binding Protein 1 (SATB1) influences the expression of multiple genes on an epigenetic level, by acting as a chromatin organizer. Importantly, SATB1 overexpression has been described several human cancers and connected to carcinogenesis. The expression of SATB1 has also been found to correlate with tumor progression and is associated with poor prognosis. The functional relevance of SATB1, its possible mechanisms of action and its potential as a target for new therapeutic strategies in colorectal cancer (CRC) were largely unknown and subject of this study. Methods: RNAi-mediated gene knockdown of SATB1 was employed in primary and in established colorectal cancer cell lines. Proliferation and colony formation assays, flow cytometry analyses of cell cycle and determination of apoptosis induction upon SATB1 knockdown were performed. By qRT-PCR and Western blotting, SATB1 knockdown effects on multiple important (proto-)oncogenes and other molecules involved in cell cycle, EMT and cell adhesion were analysed. An in vivo study using

Oncol Res Treat 2016;39(suppl 3):1–104

a nanoparticle-based platform for siRNA delivery was performed in a tumor xenograft model. Results: SATB1 knockdown led to markedly reduced SATB1 levels and caused inhibition of proliferation, deceleration of cell cycle progression and pro-apoptotic effects. Further analyses revealed effects of SATB1 on multiple signaling pathways influencing e.g. EMT, apoptosis and ErbB receptor expression. In an s.c. tumor xenograft model in athymic nude mice, stable SATB1 knockdown cells showed markedly reduced tumor formation. The therapeutic potential of SATB1 inhibition was further explored in vivo in CRC xenograft bearing mice treated with nanoparticles for siRNA delivery. A marked inhibition of tumor growth was observed in the siRNA treatment group. Conclusions: Our results indicate an important and complex role of SATB1 in the tumorigenesis of colorectal cancer. We establish SATB1 inhibition or siRNA-mediated knockdown as a promising target for pharmacological intervention. Disclosure: No conflict of interest disclosed. P302

Impact of early tumour shrinkage (ETS) on overall survival (OS) in patients with RAS wild-type (WT) metastatic colorectal cancer (mCRC) receiving first-line treatment in three randomised panitumumab trials: An exploratory study-level meta-analysis Karthaus M.1, Rivera F.2, Valladares-Ayerbes M.3, Gallego J.4, Koukakis R.5, Demonty G.6, Douillard J.-Y.7 Städtisches Klinikum München, Klinikum Neuperlach, Munich, Germany, Hospital Universitario Marqués de Valdecilla, Santander, Spain, 3Virgen del Rocio Hospital, Seville, Spain, 4Hospital General Universitario de Elche, Elche, Spain, 5Amgen Ltd, Uxbridge, United Kingdom, 6Amgen (Europe) GmbH, Zug, Switzerland, 7Institut de Cancérologie de l‘Ouest (ICO) René Gauducheau, St Herblain, France 1 2

Introduction: Tumour shrinkage is an important aim of treatment in patients with mCRC as it can provide relief of tumour-related symptoms and increase the chance of resection. ETS has been associated with improved OS and can also provide an early indication of treatment sensitivity.) A meta-analysis was conducted to analyse the impact of ETS on OS in patients with RAS WT mCRC receiving first-line treatment in three randomised panitumumab trials. Methods: Three randomised, first-line panitumumab trials have reported ETS and OS data: PRIME – phase III, panitumumab + FOLFOX4 vs FOLFOX4 (NCT00364013); PEAK – phase II, panitumumab + mFOLFOX6 vs bevacizumab + mFOLFOX6 (NCT00819780); PLANET – phase II, panitumumab + FOLFOX4 vs panitumumab + FOLFIRI (NCT00885885). This retrospective study-level meta-analysis aimed to estimate the effect of ETS ≥20% vs < 20% and ETS ≥30% vs < 30% at week 8 on OS in patients with RAS WT mCRC receiving first-line treatment (overall) in these studies. Meta-analysis techniques (fixed-effects modelling [unconditional maximum likelihood method] and random-effects modelling [DerSimonian and Laird modelling methods]), were used to pool study-level data using the inverse-variance of each trial as the weight. Results: Data from a total of 641 patients with RAS WT mCRC who were evaluable for OS and ETS in these studies, were included in the meta-analysis. Achievement of ETS ≥20% vs ETS < 20% (hazard ratio [HR]: 0.45 [95% confidence intervals [CI]: 0.37-0.54) and ETS ≥30% vs ETS < 30% (HR: 0.46 [95% CI: 0.38-0.55]) was associated with improved OS. Results from the individual studies are included in the table.

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Tab. 1.

Study PRIME (n = 440) PEAK (n = 154) PLANET (n = 47) Overall (n = 641)

HR (95% CI) for OS ETS ≥20% vs <20% 0.47 (0.38–0.58) 0.39 (0.26–0.59) 0.31 (0.11–0.83) 0.45 (0.37–0.54)

ETS ≥30% vs <30% 0.47 (0.39–0.59) 0.44 (0.30–0.65) 0.28 (0.10–0.77) 0.46 (0.38–0.55)

Conclusions: This exploratory study-level meta-analysis of pooled data from three first-line panitumumab trials (PRIME, PEAK and PLANET) suggests that ETS (≥20% or ≥30% at week 8) is associated with improved OS and hence may be a useful early indicator of treatment benefit. Disclosure: Meinolf Karthaus: Advisory Role: Consulting/advisory activities for Amgen and Roche; Financing of Scientific Research: Honoraria from Amgen and Roche; Other Financial Relationships: Travel and accomodation from Amgen and Roche Jean-Yves Douillard: Advisory Role: Consulting/advisory activities for Amgen, Bayer, Roche and Merck; Financing of Scientific Research: Honoraria from Amgen, Bayer, Roche and Merck; Expert Testimony: Research funding from Merck Serono; Other Financial Relationships: Travel, accomodation and expenses from Amgen, Bayer, Roche and Merck P303

An exploratory analysis evaluating the effect of sequence of biologic therapies on overall survival (OS) in patients (pts) with RAS wild-type (WT) metastatic colorectal carcinoma (mCRC) Karthaus M. , Sobrero A. , Douillard J.-Y. , Rivera F. , Forget F. , Valladares-Ayerbes M.6, Demonty G.7, Guan X.8, Peeters M.9 1

Städtisches Klinikum München, Klinikum Neuperlach, Munich, Germany, IRCCS Ospedale San Martino IST, Genova, Italy, 3ICO R. Gauducheau, St. Herblain, France, 4Hospital Universitario Marqués de Valdecilla, Santander, Spain, 5Centre Hospitalier de l’Ardenne, Libramont, Belgium, 6Virgen del Rocio Hospital, Seville, Spain, 7Amgen (Europe) GmbH, Medical Development, Zug, Switzerland, 8Amgen Inc., Biostatistics, Thousand Oaks, United States, 9Antwerp University Hospital, Edegem, Belgium 1 2

Introduction: Addition of epidermal growth factor receptor inhibitors (EGFRi) or vascular endothelial growth factor inhibitors (VEGFi) to chemotherapy in the first-line setting provides clinical benefit to pts with RAS WT mCRC, vs chemotherapy alone. The choice of first-line biologic therapy may impact on mCRC biology and sensitivity to subsequent treatments, as suggested by preclinical studies, but the optimal treatment sequence has not yet been assessed in a randomised prospective clinical trial. An exploratory analysis of OS for pts with RAS WT mCRC receiving either first-line panitumumab (pmab) plus second-line VEGFi, or firstline bevacizumab (beva) and second-line EGFRi was conducted. Methods: Pt-level OS data from three randomised mCRC trials was used for this exploratory retrospective analysis. PEAK (NCT00819780) was a phase II study of first-line pmab+mFOLFOX6 vs beva+mFOLFOX6; PRIME (NCT00364013) was a phase III study of first-line pmab+FOLFOX4 vs FOLFOX4 alone and study 181 (NCT00339183) was a phase III trial of second-line pmab+FOLFIRI vs FOLFIRI alone. OS was analysed for all pts with RAS WT mCRC treated with either first-line pmab (PEAK and PRIME) plus secondline VEGFi, or firstline beva (PEAK and 181) and secondline EGFRi. Results: Of the pts included in the analysis (n = 104), 66 received firstline pmab and secondline VEGFi and 38 were treated with first-line beva plus second-line EGFRi. At the time of analysis, 63.6% and 92.1% of pmab→VEGFi and beva→EGFRi pts, respectively, had died. OS HRs comparing the pmab→VEGFi and beva→EGFRi treatment sequences are reported in the table. Overall, pooled analysis of PEAK+PRIME pts vs PEAK+181 pts showed a median OS of 36.8 (95% CI: 30.3-43.8) vs 27.8 (95% CI: 24.235.6) months for pmab→VEGFi vs beva→EGFRi treatments (HR for OS [95% CI]: 0.65 [0.42-1.03]).

Abstracts

OS events, n (%) Median OS months (95% CI) HR (95% CI)

PEAK PEAK PRIME 181 Pmab→VEGFi Beva→EGFRi Pmab→VEGFi Beva→Pmab (n = 31) (n = 9) (n = 35) (n = 29) 25 (80.6)

8 (88.9)

17 (48.6)

27 (93.1)

36.8 (22.9–46.1)

25.3 (6.0–32.5)

37.4 (29.2–not estimable)

32.4 (24.2-–39.8)

0.59 (0.26-1.35)

0.65 (0.34-1.23)

Conclusions: This exploratory analysis may suggest a trend towards improved OS for pts with RAS WT mCRC treated with first-line pmab+chemotherapy followed by second-line VEGFi vs those treated with firstline beva plus second-line EGFRi, although numbers were small. A larger, prospective randomised trial is needed to confirm the appropriate sequence of EGFRi/VEGFi in these pts. Disclosure: Meinolf Karthaus: Advisory Role: Consulting/advisory activities for Amgen and Roche; Financing of Scientific Research: Honoraria from Amgen and Roche; Other Financial Relationships: Travel and accommodation from Amgen and Roche Marc Peeters: Advisory Role: Consultancy/advisory roles for Amgen; consultancy/ symposia participation for Merck Serono; Expert Testimony: Research Funding from Amgen P304

Regorafenib in previously treated metastatic colorectal cancer (mCRC): Analysis of age subgroups in the open-label phase 3b CONSIGN trial Kasper S.1, van Cutsem E.2, Ciardello F.3, Ychou M.4, Seitz J.-F.5, Hofheinz R.D.6, Arriaga Y.E.7, Verma U.7, Garcia-Carbonero R.8, Grothey A.9, Miriyala A.10, Kalmus J.10, Kappeler C.10, Falcone A.11, Zaniboni A.12 Universitätsklinikum Essen, Westdeutsches Tumorzentrum, Essen, Germany, University Hospital Leuven, Leuven, Belgium, 3Second University of Naples, Naples, Italy, 4Institut Regional du Cancer de Montpellier, Montpellier, France, 5 Aix-Marseille University, Marseille, France, 6Universitätsklinik Mannheim, Mannheim, Germany, 7University of Texas Southwestern Medical Center, Dallas, United States, 8Hospital Universitario Doce de Octubre, Madrid, Spain, 9 Mayo Clinic, Rochester, United States, 10Bayer Pharma AG, Berlin, Germany, 11 University of Pisa, Pisa, Italy, 12Fondazione Poliambulanza, Brescia, Italy 1 2

Introduction: In the phase 3 CORRECT trial, regorafenib improved survival vs placebo in treatment-refractory mCRC. The large, single-arm, phase 3b CONSIGN trial (NCT01538680) was designed to provide continued access to regorafenib for patients with mCRC and to further characterize regorafenib safety. In CONSIGN, adverse events (AEs) and progression-free survival (PFS) were consistent with results reported in phase 3 trials. We performed a subgroup analysis of CONSIGN to assess outcomes by age. Methods: Patients with mCRC who progressed after standard therapies and had ECOG PS 0─1 received regorafenib 160 mg daily for the first 3 weeks of each 4-week cycle until disease progression, death, or unacceptable toxicity. The primary endpoint was safety. PFS, assessed per investigator, was the only efficacy measure collected. Results: A total of 2872 patients were assigned to treatment (< 65: n = 1720; ≥65: n = 1152); n = 1713 and n = 1151, respectively, were evaluable for safety. ECOG PS 0/1 were 49%/51% (< 65) and 44%/55% (≥65). The median (range) treatment duration was 2.5 months (0.03–30.4) and 2.3 months (0.03–28.5) in patients < 65 and ≥65, respectively; mean (SD) percent of planned dose was 76% (20) and 74% (20), respectively. Most patients experienced a regorafenib-related AE (< 65: 91%; ≥65: 92%; Table). AEs led to treatment discontinuation in 25% (< 65) and 26% (≥65) of patients. Treatment-emergent NCI-CTCAE v4.0 hepatic grade ≥3 laboratory toxicities (< 65; ≥65) included increased bilirubin (14%; 12%), increased AST (8%; 5%), and increased ALT (7%; 4%). Estimated median

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PFS (95% CI) in the < 65 group was 2.7 months (2.6, 2.8) and in the ≥65 group was 2.6 months (2.5, 2.7).

Tab. 1.

age

Tab. 1. Drug-related AE occurring in ≥5% of patients

Drug-related AEs, n (%) Grade ≥3 Hypertension HFSR Fatigue Diarrhea Hypophosphatemia Grade 5 Serious

<65 years (n = 1713) 939 (55) 232 (14) 274 (16) 180 (11) 196 (17) 78 (5) 89 (5) 8 (0.5) 151 (9)

57 (5) 60 (5) 5 (0.4) 100 (9)

Conclusions: The safety profile of regorafenib was generally comparable in patients <65 and ≥65 years of age. PFS was similar across age subgroups. Disclosure: Stefan Kasper: Advisory Role: Amgen, Bayer, BMS, Celgene, Lilly, Merck, MSD, Roche, Sanofi.; Financing of Scientific Research: Amgen, Lilly, Merck, Sanofi; Expert Testimony: Celgene, Merck; Other Financial Relationships: Amgen, Lilly, Meck, Sanofi, Roche, Bayer (Travel) Alberto Zaniboni: No conflict of interest disclosed. P305

>65–75 yrs (b) n = 155 48.6 53.0 10.1 10.4 5.2 4.1*

> 75 yrs (c) n = 66 50.0 21.1* 9.2 7.8 5.5 2.8*

Total n = 456 51.8 “ 11.5 “ 5.1 “

*p < 0.05 (age group ≤ 65 used as control); a CCI 0 & ≥1: n = 148 & 87; b CCI 0 & ≥1: n = 72 & 83; cCCI 0 & ≥1: n = 28 & 38+

Disclosure: Stephan Sahm: Stock Ownership: Fresenius Medical Care; Merck KGaA; Financing of Scientific Research: Fresenius Medical Care; Expert Testimony: Merck Serono (Institution); Roche (Institution) Friedrich Overkamp: Advisory Role: Merck Serono; Financing of Scientific Research: Merck Serono P306

The colorectal carcinoma – treatment research and treatment reality in oncology practices (Anti-VEGF or Anti-EGFR therapies) Tessen H.-W.1, Rubanov O.2, Grundeis M.3, Teich M.3, Elsel W.4, Schlichting A.5, Valdix A.6, Projektgruppe Internistische Onkologie (PIO) Onkologische Kooperation Harz, Goslar, Germany, 2Onkologische Schwerpunktpraxis, Hameln, Germany, 3Onkologische Schwerpunktpraxis, Chemnitz, Germany, 4Onkologische Schwerpunktpraxis, Glauchau, Germany, 5 rgb Onkologisches Management GmbH, Sarstedt, Germany, 6Onkologische Schwerpunktpraxis, Schwerin, Germany 1

Outcome of patients with KRAS exon 2 wildtype (KRAS-wt) metastatic colorectal carcinoma (mCRC) with cetuximabbased first-line treatment in the non-interventional study ERBITAG and impact of comorbidity and age Sahm S.1, Göhler T.2, Hering-Schubert C.3, Janssen J.4, Neumann U.P.5, Schwittay M.6, Zahn M.-O.7, Stenzel K.G.8, Steinbach-Büchert A.K.8, Overkamp F.9 Ketteler Krankenhaus, Offenbach, Germany, 2Onkozentrum Dresden/Freiberg, Dresden, Germany, 3St. Georg Klinikum, Eisenach, Germany, 4Onkologie Westerstede Aurich Rhauderfehn, Westerstede, Germany, 5Universitätsklinik der RWTH, Aachen, Germany, 6Tumorzentrum Leipziger Land, Groitzsch, Germany, 7 Onkologische Kooperation Harz, Goslar, Germany, 8Merck Serono GmbH, Darmstadt, Germany, 9Oncologianova GmbH, Recklinghausen, Germany 1

Background: Cetuximab in combination with irinotecan- or oxaliplatin-based chemotherapy has shown to increase ORR, PFS, OS of KRASwt mCRC patients (pts). ERBITAG aimed to evaluate safety and efficacy of cetuximab in combination with various first-line chemotherapy regimens in pts with unresectable KRAS-wt mCRC. Methods: KRAS-wt pts on a cetuximab-based first-line treatment with written informed consent could be enrolled in this prospective, non-interventional study. Primary endpoint was ORR, secondary endpoints were amongst others PFS, OS, TTF, and resection rate of liver metastasis. Comorbidities were documented and evaluated by the Charlson Comorbidity Index (CCI). Results: 817 eligible KRAS-wt mCRC pts were enrolled at 144 sites across Germany, documentations for 456 pts were finalised and evaluated. The median age was 65 [27–87] yrs, with 51.5% ≤ 65 yrs, 34.0% > 65–75 yrs, and 14.5% > 75 yrs. ECOG performance status was 0, 1, 2, or missing in 34.4%, 49.6%, 8.8%, and 7.2% of pts, respectively. CCI was 0 in 54.4%, and ≥ 1 in 45.6%. Resection of liver and/or lung metastases was done in 17.3% of pts, 13.4% were R0 resected. For pts with liver limited disease resection rate and R0-rate were 29.3% and 23.8%, respectively. Pts with CCI 0 had no different outcome regardless of age (Tab.1). Pts with CCI ≥ 1 and > 75 yrs had a lower ORR and decreased TTF, pts > 65–75 yrs had only a decreased TTF as compared to the ≤ 65 yrs age group (Tab. 1). Conclusions: In this large observational trial outcomes (ORR and PFS) of KRAS-wt mCRC pts on a cetuximab-based first-line treatment were comparable to those reported in pivotal trials. Pts older than 75 yrs without comorbidities (CCI = 0) showed no difference to younger pts in ORR, PFS, and TTF. Pts > 75 yrs with CCI ≥ 1 had a significant lower ORR and decreased TTF.

CCI = 0 ORR % CCI ≥ 1 ORR % CCI = 0 PFS mo. CCI ≥ 1 PFS mo. CCI = 0 TTF mo. CCI ≥ 1 TTF mo.

≥65 years (n = 1151) 690 (60) 203 (18) 122 (11)

≤65 yrs (a) n = 235 60.1 52.9 13.6 9.6 5.3 6.0

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Introduction: For the palliative therapy of the colorectal carcinoma there are potent drugs available such as the cytostatics 5-FU, irinotecan, oxaliplatin and capecitabine. In 01/2005, the monoclonal antibody bevacizumab was approved, cetuximab in 06/2004, panitumumab in 12/2007 (cetuximab and panitumumab for carcinoma with wild-type Ras). As of 02/2013, aflibercept can be administered in combination with FOLFIRI after an oxaliplatin-based treatment. How are these substances applied / combined in everyday life? Methods: Since 2003, 124 oncology practices in 16 federal states have been documenting 9,301 disease histories of patients with a colorectal carcinoma (CRC) as part of the Project team of Internal Oncology (PIO). 8,687 cases thereof with a total of 19,589 therapies were analysed in the registry ONCOReg. Distant metastases were present in 5,691 patients during the course of the disease. Results: For 5,606 patients, 14,053 palliative therapies have been documented so far. Tab. 1. Therapies

Patients Therapies 1st-line (%) 2nd-line (%) 3rd-line (%)

Total group 5,606 14,053 100.0 67.7 37.6

Bevacizumab 3,238 4,674 73.8 34.9 15.9

Cetuximab 1,752 2,132 25.4 47.6 30.2

Panitumumab 630 680 9.0 15.1 30.0

Aflibercept 217 233 2.8 31.8 25.8

1,026 (18.0%) of the patients had a secondary metastases resection, 2.5% of the patients an interventional treatment for metastases. Tab. 2. Secondary metastases resection

Bevacizumab Cetuximab n = 3,170 n = 1,715

Panitumumab n = 623

Aflibercept n = 217

Secondary metastases resection

574 (18.1%)

144 (23.1%)

69 (31.8%)

301 (17.6%)

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CONTENTS AUTHOR INDEX

The median progression-free survival in the complete registry is at 9.9 or 6.4 months for the 1st- and 2nd-line therapy, the median overall survival at 25.0 and 15.3 months. Patients with a secondary metastases resection survived 50.5 months, patients without a secondary metastases resection 24.8 months (p < 0.00001). Patients who were treated with aflibercept and had received a metastases resection survived 54.5 months. Conclusion: The median overall survival of 25.0 months from the start of the 1st-line therapy reflects the targeted treatment of the colorectal carcinoma in the practices. Patients with a secondary metastases resection had a significantly prolonged overall survival (54.5 months with an aflibercept-containing therapy). Current data will be presented. Disclosure: No conflict of interest disclosed. P307

Treatment and outcome of patients with metastatic colorectal cancer (mCRC) in routine care 1995 - 2015 Weide R.1, Feiten S.2, Chakupurakal G.1, Friesenhahn V.2, Kleboth K.2, Köppler H.1, Lutschkin J.2, Thomalla J.1, van Roye C.1, Heymanns J.1 Praxisklinik für Hämatologie und Onkologie, Koblenz, Germany, 2Institut für Versorgungsforschung in der Onkologie, Koblenz, Germany 1

Introduction: Evaluation of treatment strategies and outcome of patients with mCRC who received their treatment in an oncology group practice by 5 oncologists between 06/95 - 04/15. Comparison of different treatment periods (1995 - 2004 versus 2005 - 2015). Methods: All consecutive patients with mCRC who were diagnosed between 06/95 - 04/15 were analysed retrospectively concerning treatment and outcome. Data were collected from patient files into a data base and analysed statistically using SPSS. Results: 526 patients were analysed. Median age was 66 (28 - 89). 45% were female, 55% were male. KRAS-mutation status was known in 25%; 44% were mutated, 56% showed a KRAS-wildtype. In patients diagnosed between 2007 and 2015 (n = 148) KRAS-mutation status was known in 75%; 48% were mutated, 52% wildtype. First line therapy consisted of 5-FU+folinic acid-regimens in 75%; FOLFIRI was applied in 14%, FOLFOX in 25%, CAPIRI in 2% and CAPOX in 4%. Bevacizumab was used in 17%, an Anti-EGFR-antibody in 1%. Second line therapy consisted of 5-FU+folinic acid-regimens in 64%; FOLFIRI was applied in 21%, FOLFOX in 28%, CAPIRI in 2% and CAPOX in 7%. Bevacizumab was used in 22%, an Anti-EGFR-antibody in 4%. Third line therapy consisted of 5-FU+folinic acid-regimens in 44%; FOLFIRI was applied in 23%, FOLFOX in 15%, CAPIRI in 7% and CAPOX in 6%. Bevacizumab was used in 18%, an Anti-EGFR-antibody was used in 18% as well. 18 patients (3%) received regorafenib as third and further line therapy. 4% of patients received best supportive care only. The median overall survival is 23.5 months (1.4 - 193.4+). In patients >75 years median overall survival is 22.6 months (2.8 - 163.1+). The comparison of different treatment periods (1995 - 2004 versus 2005 - 2015) revealed a substantial difference in survival (22.3 months versus 27.8 months). Conclusion: Modern cytoreductive therapy leads to an improvement in overall survival in patients with mCRC who receive routine care. Substantial improvement has been achieved during the last ten years compared to 1995 - 2004.

P308

Impact of surgical resection of liver metastases on outcome of patients with KRAS-wildtype exon 2 (KRAS-wt) metastatic colorectal carcinoma (mCRC) treated with a cetuximab-based first-line therapy – Analysis of survival times in relation to secondary resection in the German non-interventional study ERBITAG Neumann U.P.1, Göhler T.2, Hering-Schubert C.3, Janssen J.4, Sahm S.5, Schwittay M.6, Zahn M.-O.7, Stenzel K.G.8, Overkamp F.9 Universitätsklinik der RWTH Aachen, Klinik für Allgemein-, Viszeral- und Transplantationschirurgie, Aachen, Germany, 2Onkozentrum Dresden/ Freiberg, Dresden, Germany, 3St. Georg Klinikum Eisenach, Eisenach, Germany, 4 Onkologie Westerstede Aurich Rhauderfehn, Westerstede, Germany, 5Ketteler Krankenhaus, Offenbach, Germany, 6Tumorzentrum Leipziger Land, Groitzsch, Germany, 7Onkologische Kooperation Harz, Goslar, Germany, 8Merck Serono GmbH, Darmstadt, Germany, 9Oncologianova GmbH, Recklinghausen, Germany 1

Background: Cetuximab in combination with irinotecan- or oxaliplatin-based chemotherapy has shown to increase amongst other efficacy parameter the R0 resection rate of liver metastases (LM) of primary irresectable KRAS-wt mCRC patients (pts). The non-interventional study ERBITAG aimed to evaluate safety and efficacy of cetuximab in combination with various first-line chemotherapy regimens in pts with unresectable KRAS-wt mCRC. Methods: KRAS-wt pts on a cetuximab-based first-line treatment with written informed consent could be enrolled in this prospective, non-interventional study. Primary endpoint was ORR, secondary endpoints were amongst others PFS, OS, TTF, and resection rate of liver metastasis. Here we update the interim analysis in terms of the outcome of pts according to secondary resection of LM. Results: 456 KRAS-wt mCRC pts out of 817 recruited KRAS-wt pts were evaluable for this interim analysis. Location of metastasis was liver, lung, lymph nodes, and peritoneum in 70.6%, 23.5%, 22.4%, and 17.8% of pts, respectively. 39.7% of pts had liver-limited disease (LLD). 79.2% had surgery of the primary tumor and 15.6% had a resection of metastases before enrolment. The median [range] age was 65 [27–87] yrs, ECOG performance status was 0, 1, 2, or missing in 34.4%, 49.6%, 8.8%, and 7.2% of pts, respectively. Resection of LM and/or lung metastases was done in 17.3% of pts, 13.4% were R0 resected. For LLD-pts resection rate and R0rate were 29.3% and 23.8%, respectively. Intraoperative and postoperative complications were observed in 1.1% and 12.4% of pts with resection of metastasis, respectively. In 4.5% of pts a revision surgery was necessary. Median PFS was 20.3 and 9.5 months for pts with and without resection of LM, respectively (p < 0.0001). Median OS was 42.0 and 18.2 months for pts with and without resection of LM, respectively (p < 0.0001). For LLDpts median PFS was 20.5 and 11.9 months and median OS 42.0 and 18.7 months for pts with and without resection of LM, respectively(p < 0.0001 each). Conclusions: Pts with resection of LM had a significant longer PFS and OS compared to pts without resection. Disclosure: Ulf Neumann: No conflict of interest disclosed. Friedrich Overkamp: Advisory Role: Merck Serono; Financing of Scientific Research: Merck Serono

Disclosure: No conflict of interest disclosed.

Abstracts

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P309

P310

Mutation profiling and consecutive treatment decisions under knowledge of mutation results in patients with metastatic colorectal cancer (mCRC) in real world evidence (RWE) of oncological practices – first results of the GO-KOLORAS registry study

Impact of prophylactic treatments of cetuximab-based skin reactions in patients with metastatic colorectal carcinoma (mCRC). Interim analysis of the German non-interventional study ERBITAG

Lipp R. , Freigang F. , Brecht P. , Schwaner I. , Steinmetz T. , Schulte C. , Tiemann M.5 1

GermanOncology GmbH, Hamburg, Germany, 2Onkologische Schwerpunktpraxis Kurfürstendamm, Berlin, Germany, 3Gemeinschaftspraxis für Hämatologie und Onkologie, Köln, Germany, 4Gemeinschaftspraxis für Hämatologie und Onkologie, Dortmund, Germany, 5Institut für Hämatopathologie, Hamburg, Germany 1

Introduction: The GO-KOLORAS registry study started in October 2015 and investigate the mutation profilings of several markers in tumor samples of colorectal cancer analysed by Next-Generation-Sequencing (NGS) and the subsequent decisions of treatments for 1° and 2°line based on the mutation reports under conditions of RWE in German oncological practices. Methods: From October 2015 data of the first n = 78 (of planned 1.440 patients) with mCRC were sampled from more than 20 oncological practices in Germany. The NGS examinations of KRAS, NRAS, AKT 1, APC, BRAF, CTNNB1, PIK3CA, PTEN, SMAD4, TP53 and MSS were performed centrally in formalin-fixed paraffin-embedded tumor samples and reported to the oncologists at the point of a decision for a 1° or 2°line. All data of mutation profilings and clinical as well as health economic data were captured by a web-based documentation software. Clinical data of the GO-KOLORAS trial were compared with historical data of 342 patients with mCRC, who started 1° or 2°line therapies in the same oncological practices between 2014 and 2015. Results: Of the first patients with mCRC in the GO-KOLORAS trial (34% women, 66% men, mean age 68 years) the NGS analyses showed the following portions of mutations: KRAS 40,0%, NRAS 6,7%, AKT 1 2,2%, APC 80,0%, BRAF 8,9%, CTNNB1 2,2%, PIK3CA 20,0%, PTEN 4,4%, SMAD4 20,0%, TP53 80,0%. Additionally we found an instability in MSS in 2,2%. The total rate of RAS wildtype was 53,3% (in subgroups WT for KRAS 60,0%, for NRAS 93,3%). In the WT group (1° and 2°line) the portions of targeted therapies with Cetuximab/Panitumumab could be increased by 9,3% in the 1° and by 7,2% in the 2°line compared to historical data from 2014–2015. Updated data regarding the treatments and analyses of marker profiling groups will be presented at the meeting. Conclusions: NGS with a broad marker profiling could provide essential informations about RAS status and additional profilings regarding further markers. The GO-KOLORAS registry will investigate the subsequent decisions of the oncologists for 1° and 2°line therapies in mCRC based on the NGS reports with a broad marker profiling under conditions of RWE. First data showed a slight increase of targeted therapies in 1° and 2° line, which may be caused in the contemporary submission of the NGS results, but up to now no significant difference to the historical data from 2014–2015. Disclosure: Rainer Lipp: Advisory Role: Beratertätigkeiten bei Omnicare; Financing of Scientific Research: Vortragshonorar von Omnicare; Expert Testimony: Forschungsunterstützung von Omnicare Markus Tiemann: No conflict of interest disclosed.

Oncol Res Treat 2016;39(suppl 3):1–104

Sahm S.1, Göhler T.2, Hering-Schubert C.3, Janssen J.4, Schwittay M.5, Zahn M.-O.6, Stenzel K.G.7, Steinbach-Büchert A.K.7, Overkamp F.8 Ketteler Krankenhaus, Offenbach, Germany, 2Onkozentrum Dresden/ Freiberg, Dresden, Germany, 3St. Georg Klinikum Eisenach, Eisenach, Germany, 4Onkologie Westerstede Aurich Rhauderfehn, Westerstede, Germany, 5Tumorzentrum Leipziger Land, Groitzsch, Germany, 6Onkologische Kooperation Harz, Goslar, Germany, 7Merck Serono GmbH, Darmstadt, Germany, 8 Oncologianova GmbH, Recklinghausen, Germany 1

Background: Prophylactic treatments are widely used to ameliorate skin reactions induced by EGFR inhibition. Randomised studies have shown a positive impact of prophylactic antibiotics. Methods: Patients (Pts) with KRAS-wt and later RAS-wt mCRC treated with a first-line chemotherapy regimen plus cetuximab with written informed consent were eligible for this prospective, non-interventional study. Physicians were requested to complete a questionnaire and document any applied prophylactic and reactive skin toxicity treatment for every pt. Different prophylactic treatment regimens were categorized in 5 groups: systemic antibiotics (SA), skin care without antibiotics or corticosteroids (SC), other topical treatments (OT) (e.g. antibiotics or corticosteroids), any prophylaxis (P), and no prophylaxis (NP). Results: Data from 497 pts at 178 centers were finally collected and evaluable at data cut off. For all reported skin reactions the maximum NCI-CTCAE grade per patient and prophylactic treatment group was evaluated (table). Although none of the prophylactic regimens were significant different from NP, SA showed a numerically lower rate of all skin reactions by 8.1% (p = 0.06) and of rash acneiforme by 5.3% (p = 0.134) versus NP. 96.2% of reactive treatments of skin toxicities were done without consultation of a dermatologist and the medication was topical or systemic in 60.9% and 32.3%, respectively. Response to reactive treatment was complete remission or significant improvement of the skin reactions in 68.6% (topical) and 66.0% (systemic). Conclusions: Pts given prophylactic SA showed numerically fewer grade 3–4 skin reactions in comparison to NP (7.6% vs. 15.7%), but without significance. An important reason for failing significance may be the low number of pts receiving prophylactic SA (only in 18.5% of pts). Reactive treatment (systemic or topical) of skin toxicities led to an improvement in the majority of pts. Tab. 1. Skin reactions with different prophylaxis n = 497 NP n = 198 (39.8%) P n = 299 (60.2%) SA n = 92 (18.5%) SC n = 37 (7.5%) OT n = 170 (34.2%)

all skin reactions grade 3/4 [%] 15.7 11.4 7.6 18.9 11.8

rash acneiforme grade 3/4 [%] 8.6 6.4 3.3 13.5 6.5

Abstracts

CONTENTS AUTHOR INDEX

P311

Simultaneous diagnosis of aggressive B-NHL and coloncancer: Case report and review of the literature Chitic A.P.1, Sprey C.1, Meyer R.1, Schulte E.1, Hindahl H.1, Bolder U.1, Luckhaupt H.1 St Johannes Hospital Dortmund, Department of Internal Medicine II, Department of Surgery and Department of ENT, Dortmund, Germany 1

Introduction: Treating patients according to guidelines is standard of care. However, the simultaneous diagnosis of two malignant diseases usually is not covered by these guidelines. We report on an older patient, diagnosed with B-NHL and colon cancer. Case: A 85 years old woman, presented with intermittent abdominal pain and obstipation. She was in very good performance status (ECOG 1) without relevant co-morbidities besides a known hypercholesterinemia. Upon clinical examination, our patient revealed moderate pain in the right lower abdominal quadrant, without muscular defense. An abdominal ultrasound showed splenomegaly and para-aortic lymphadenopathy. CT-scan confirmed splenomegaly and generalized lymphadenopathy. In addition, an infiltrative process of the coequal region was found. Colonoscopy exhibited a stenosing ulcerative mass in the cecum, with the typical histological features of a colon adenocarcinoma. A supraclavicular lymph node was surgically extracted, and a diffuse large B cell lymphoma was histologically confirmed. Bone marrow involvement was ruled out by flow cytometry and histology. While having extended evidence supported guidelines and therapy algorithms for both these pathologies, none of them provides recommendations for cases with simultaneous tumors of different histology. In addition, the enlarged abdominal lymph-nodes could have been manifestations of either disease. After discussion in our local tumorboard and intensive discussions with the patient and her family, we decided treating the lymphoma first, and initiated a therapy with elderly R-CHOP-14. This therapy was very well tolerated and a restaging after four cycles chemotherapy documented not only regression of all lymphoma manifestations but also of the cecal mass. After six cycles, the patient was in complete remission with the lymphoma. She subsequently underwent surgical resection of the colon carcinoma in curative intention. Conclusion: A better documentation in a registry of these patients with two oncological diseases would help us to observe how other colleges in such a situation are handling. In our patient with a stage III A Lymphoma and a curative treatable colon carcinoma (postoperative stage II UICC), was this strategy the only chance to cure the patient. An interesting side effect is the partially response of the colon cancer under lymphoma therapy.

ciplinary treatment, developing individual therapeutic strategies remains a challenge. There is an ongoing need for developing assays to predict treatment response of individual patients and tumours. Methods: Our group succeeded to cultivate tissue slices from primary gastric carcinomas (Koerfer J et al., 2016). To implement this model for clinical applications a fast and objective readout is necessary. We therefore standardised the established slice culture system for gastric and colon cancer and optimised a histology based readout. A triplicate slice approach was used to address the problem of tumour heterogeneity. Cytotoxic drugs (oxaliplatin, cisplatin, 5-FU, docetaxel) were added to culture medium for 48–72 hours after a resting period. Analysis was done using ImageJ (ImageJ, Schneider et al., 2012). In short, paraffin sections were prepared and stained with different immunofluorescence dyes such as Hoechst 33342, Ki67 and AE 1+3. Images of three slices were pooled and modified with a standardised, automated workflow in ImageJ. The mean value of positive pixels per condition was calculated and results were correlated with manual cell counting and experimental conditions. Results: Our standardised experimental setup in combination with an automated, pixel-based readout represents a promising option to analyse human tumour derived tissue of gastrointestinal cancers. Values obtained by using the ImageJ workflow correlated with manual cell counting of four observers (n = 48): r = 0.841 and no significant difference was found between normed values of both methods within observers (n = 12): p = 0.885, p = 0.885, p = 0.977and p = 0.707. Conclusions: We report results of our standardised slice culture readout system to assess response of human gastric and colon cancer tissue to chemotherapy ex vivo. This novel method offers the opportunity to study tumour biology and drug resistance within a model closer to its original environment compared with other models. It is a first step towards an assay that might enable the prediction of individual tumour response prior to therapy. Disclosure: Rasmus Soennichsen: No conflict of interest disclosed. Florian Lordick: Advisory Role: Amgen, Biontech, Boston Biomedical, Ganymed, Lilly, MSD, Nordic, Roche, Taiho; Financing of Scientific Research: Vortragshonorare: Amgen, Celgene, Roche, Lilly; Expert Testimony: Böhringer-Ingelheim, GSK, Fresenius Biotech; Immaterial Conflict of Interests: Reisekostenunterstützung: Amgen, Bayer, MSD, Roche, Taiho

Fortbildung

Myelodysplastisches Syndrom V313

Disclosure: No conflict of interest disclosed.

WHO classification 2016 for the myelodysplastic syndromes (MDS): Meaning for the diagnostic work-up

P312

Germing U.1, Strupp C.1, Nachtkamp K.1, Aul C.2, Giagounidis A.3, Gattermann N.1

Development of a standardised and reliable slice culture system for gastrointestinal cancers Soennichsen R.1,2, Kallendrusch S.1, Hennig L.1,2, Koerfer J.1,2, Merz F.1, Winter K.1, Haehnel S.1, Kaiser N.1, Richter C.1, Monecke A.3, Wittekind C.3, Hoffmeister A.4, Jansen-Winkeln B.5, Gockel I.5, Aigner A.6, Bechmann I.1, Lordick F.2

Heinrich-Heine University Düsseldorf, Department of Hematology, Oncology and Clinical Immunology, Düsseldorf, Germany, 2St Johannes Hospital Duisburg, Department of Hematology, Oncology and Clinical Immunology, Duisburg, Germany, 3Marienhospital Düsseldorf, Department of Hematology and Oncology, Düsseldorf, Germany 1

Introduction: Gastric and colon cancers are among the most prevalent malignant tumours and have a high mortality rate. Gastric cancer in particular is characterized by heterogeneity, low overall survival and high rates of non-responders. Despite recent advances in medical and multidis-

A refinement of the classification of the MDS has been proposed by the WHO working group in 2016: 1) definition of the ring sideroblastic MDS types including SF3B1 mutation as a classifier for patients with less than 15% ring sideroblasts (RS) and the reintroduction of a group of MDS with multilineage dysplasia and RS, 2) inclusion of MDS with del(5q) and one additional non chromosome 7 anomaly in to the MDS del(5q) group, 3) replacement of the term RCUD by MDS with single lineage dysplasia (MDSSLD), RARS by MDSSLDRS (with Ringsideroblasts), RCMD by MDS multilineage dysplasia (MDSMLD), and MDSMLDRS, RAEB by MDSEB, and 4) definition to assess the medullary blast count, namely the blasts should be based on all nucleated cells neglecting the amount of erythroid cells. Based on centrally diagnosed 3190 patients in the Düsseldorf registry, we validated the new proposals. 256 patients were diagnosed

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–104

University of Leipzig, Institute of Anatomy, Leipzig, Germany, University Hospital Leipzig, University Cancer Center Leipzig, Leipzig, Germany, 3University Hospital Leipzig, Institute of Pathology, Leipzig, Germany, 4University Hospital Leipzig, Department of Gastroenterology and Rheumatology, Section of Interdisciplinary Endoscopy and Sonography, Leipzig, Germany, 5University Hospital Leipzig, Department of Visceral, Transplant, Thoracic and Vascular Surgery, Leipzig, Germany, 6University of Leipzig, Rudolf-Boehm-Institute for Pharmacology and Toxicology, Clinical Pharmacology, Leipzig, Germany 1

CONTENTS AUTHOR INDEX

as MDSSLD (8%), 978 MDSMLD (30,6%), 227 MDSSLD RS (7,1%); 321 MDSMLD RS (10,1%), 159 MDS del(5q) (5%), 481 MDSEB 1 (15,1%), 620 MDSEB 2 (19,5%), and 148 MDS-U (4,6%). 344 patients (16,5% of the non RAEB types) changed the category, mainly moving from RCMD to MDSMLD-RS, RCUD and RCMD to MDS del(5q). Median survival times of the refined groups differed from more than 63 months in the MDSSLD (RS) groups, 36 months in the MDSMLD (RS) groups, 76 months of the MDS del(5q) group and 21 and 11 months in the MDSEB 1 and 2 groups, respectively. The difference between the groups with regard to the risk of AML evolution also differed significantly. No major changes were made with regard to the MDS-U categories, including patients with MDSSLD and pancytopenia, MDSSLD and MDSMLD with 1% peripheral blasts and MDS without clear dysplasia but specific chromosomal aberration. In summary, the proposals of the WHO group for the classification of MDS are thoughtful, taking into account biologic parameters of the diseases, a more precise wording, to some extend pragmatic and feasible. Disclosure: No conflict of interest disclosed. V314

MDS: Standards of therapy in 2016 Platzbecker U.1, Deutsche MDS und EMSCO Universitätsklinikum Dresden, Medizinische Klinik und Poliklinik I, Dresden, Germany 1

The heterogeneity of MDS manifests in the individual patient as a disease ranging from an indolent condition with a considerable life expectancy to forms approaching the aggressiveness of acute myeloid leukemia (AML). A risk-adapted treatment strategy is therefore mandatory for a disease showing such a highly variable clinical course. During the past decades treatment has been stratified according to the International Prognostic Scoring System (IPSS) risk score; i.e. into “lower-risk” MDS (low/ int-1, LR-MDS) where correction of cytopenia was the main objective and “higher-risk” MDS (int-2/high, HR-MDS) where the reduction or delay of progression or AML evolution and prolonged survival was the objective. Erythropoiesis-stimulating agents (ESAs) constitute the first line treatment except for transfusion dependent patients with a 5q deletion, where lenalidomide can induce higher response rates. Furthermore, results of second line treatments of anemia are modest, and most of the patients will eventually require repeated RBC transfusions, subsequently leading to iron overload. In this situation, the role of iron chelation therapy requires further studies, but it is usually recommended in highly transfused patients with an expected prolonged survival or when scheduled for allogeneic HSCT. In case of thrombocytopenia, TPO-R agonists might be effective, while HMAs may be useful in selected cases. Intensive treatment approaches are hampered by the advanced age of most MDS patients, which potentially results in an unacceptable adverse event rate. This is especially true for allogeneic hematopoietic stem cell transplantation, which, despite curative potential, is selected as an option by only a minority of patients. Recent developments with hypomethylating agents have broadened the therapeutic armamentarium and these newer treatments, although not potentially curative, can lead to durable responses in many MDS patients. Most importantly, new treatment approaches are currently explored inside clinical trials including inhibitors of the TGF-beta pathway and combinations of IMIDs, ESAs or other epigenetic drugs with HMAs. In the interest of our patients we hope that these efforts will extend our therapeutic armamentarium in the near future. Disclosure: Uwe Platzbecker: Advisory Role: Celgene, Amgen, Janssen, Novartis; Financing of Scientific Research: Celgene, Amgen, Janssen, Novartis; Expert Testimony: Celgene, Amgen, Janssen, Novartis

Oncol Res Treat 2016;39(suppl 3):1–104

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When hypomethylating agents fail Pfeilstöcker M.1 Hanusch Hospital, 3rd Dept Medicine, Vienna, Austria

Introduction: Hypomethylating Agents (HMA) are treatment standard for high risk MDS patients not eligible for allogeneic transplantation. Clinical benefit and improved overall survival (OS) was shown in trials and registries. However up to 40% of patients do not respond and most responders progress beyond 1–2 years. Outcome after HMA failure is poor. Methods: Possible strategies addressing HMA failure are reviewed. Results: Restarting HMA in former responders has dismal outcomes, therefore HMA should be maintained in responders. Data on subsequent treatment with Decitabine after Azacytidine (Aza) failure are limited, showing only short OS. New HMAs currently being tested are Guadecitabine and Sapacitabine. Standard chemotherapy after HMA failure is not recommended, first line data in MDS and second line use in AML show inferior results. Newer chemotherapy agents such as Vosaroxazin might provide benefit in elderly patients, a trial in high risk MDS is ongoing. Genetically heterogeneous MDS provides a plethora of potential targets for treatment approaches: among others early data with immune checkpoint inhibitors and modulators of apoptosis merit further study. Randomized phase 3 data are scarce, only the ras mimetic Rigosertib was investigated after HMA failure randomized against best supportive care. The primary endpoint OS was not met, but benefit in primary HMA failure and very high risk patients was seen, a respective confirmatory trial is ongoing. Studies of combination treatment with the aim of improving monotherapy results have not yet been successful in phase 3. A trial testing Aza in combination with Lenalidomide or Vorinostat was negative, due to increased toxicity and failure to deliver full planned doses. For further progress in this field identification of druggable targets common to all or many MDS patients would be beneficial: the splicing apparatus could represent a new target option. Predictive markers for specific agents need to be identified, also using molecular data from NGS involving computer models. Probably also the design of clinical trials needs to be reconsidered. Conclusions: There is still need for improvement in HR MDS therapy where no standard approach for the relapsed setting is available, therefore patients should be included in clinical trials whenever possible. Those that become eligible for transplant with first line treatment should be reevaluated in this respect, as this still represents the only curative treatment option. Disclosure: Michael Pfeilstöcker: Financing of Scientific Research: Celgene, Janssen, Novartis

Fortbildung

Chronische lymphatische Leukämie V319

Treatment failure: toxicities, clonal evolution and Richter´s transformation Steurer M.1 Medizinische Universität, Innsbruck, Austria

Treatment failure in oncology has often been reduced to drug resistance of the malignant clone. However, other aspects such as toxicity may also limit the antineoplastic activity of a specific treatment modality. In the field of CLL potent immuno-chemotherapeutic regimens and the recent advent of novel agents have improved our therapeutic armamentarium but have also introduced distinct toxicities that have to be dealt with. Moreover, they may also impact on the biology of the disease in terms of clonal evolution and/or Richter´s transformation. In this educational overview three defined aspects of treatment failure, i.e. toxicities, clonal evolution and Richter´s transformation, will be discussed – in particular with focus on novel agents.

Abstracts

CONTENTS AUTHOR INDEX

Disclosure: Michael Steurer: Advisory Role: Roche, Janssen-Cilag, Gilead; Financing of Scientific Research: Roche, Janssen-Cilag, Gilead; Expert Testimony: Roche, Janssen-Cilag, Gilead

Fortbildung

Sarkome 2016 – was gibt es Neues? V321

Metastatic soft tissue sarcomas in 2016 - update and current clinical trials Kasper B.1 Universität Heidelberg, Universitätsmedizin Mannheim, Interdisziplinäres Tumorzentrum Mannheim (ITM), Sarkom Zentrum, Mannheim, Germany 1

Soft tissue sarcomas (STS) are a heterogeneous group of tumours arising mainly from the embryonic mesoderm comprising more than 50 different histological entities exhibiting great differences in terms of clinical behavior, pathogenesis and genetics. In up to 50% of patients distant metastases will occur and the median overall survival for patients with advanced disease is approximately 12 months. As effective targeted treatments are scarce, doxorubicin and ifosfamide – being used for more than 30 years now – still remain the backbone of systemic chemotherapy. In most cases, patients with advanced STS have a poor prognosis and the primary goal of treatment is disease control and palliation. Recently, trabectedin, pazopanib and eribulin have been introduced beyond first line therapy and have enriched the therapeutic armamentarium significantly. New data and clinical trials of interest will be presented here. Regarding trabectedin – approved in Europe in 2007 – a large phase III study in the USA comparing trabectedin versus DTIC stressed its sustained activity in STS patients. Even though the primary endpoint, overall survival, was not reached, the results led to global FDA registration in October 2015 for the treatment of patients with leiomyosarcomas and liposarcomas. The anti-angiogenic compound pazopanib has been tested in a large EORTC phase III trial (PALETTE) demonstrating a significant advantage regarding PFS prolongation of about three months in favour of pazopanib versus placebo for certain STS subtypes excluding liposarcomas. This study led to its approval in USA, Europe and Japan in 2012. The phase III trial of eribulin met its primary endpoint of an overall survival benefit of two months (13.5 versus 11.5 months) in favour of eribulin compared to DTIC in pretreated patients with advanced leiomyosarcomas or adipocytic sarcomas. The final approval of eribulin in early 2016, however, was restricted to pretreated liposarcoma patients. Olaratumab, a fully human anti-PDGFRα monoclonal antibody, has been tested in a phase II trial in combination with doxorubicin; olaratumab is the first agent added to doxorubicin to improve overall survival (25.0 versus 14.7 months) in advanced/metastatic STS patients in a randomized setting. A worldwide, placebo-controlled phase III study (ANNOUNCE) finalized recruitment. Another interesting candidate being tested in a phase II/III study in liposarcoma patients is selinexor, a nuclear exportin protein 1 inhibitor.

Patients with more advanced disease (either with metastasis at presentation or with recurrence) require newer approaches and clinical trials are ongoing of promising agents. Several studies focus on the introduction of a personalized medicine approach. Aim of these studies is to identify targets on the individual tumor in order to provide a personalized medicine recommendation. Pilot studies are ongoing and results are pending. Furthermore, phase II trials on PARP inhibitors, tyrosine kinase inhibitors, monoclonal antibodies, checkpoint inhibitors and other immunomodulators are tested. Results on completed studies will be presented and discussed. Disclosure: No conflict of interest disclosed. V323

Proton therapy – chances and limits? Geismar D.1,2, Timmermann B.1,2 West German Proton Therapy Center, Essen, Germany, 2Clinic for Particle Therapy, University Hospital, Essen, Germany 1

Protons are particles with a positive charge, inertia and energy depended range in tissue. In comparison to photons, protons have totally different physical properties. Energy output of protons after entry into the tissue is low at first and only rises abruptly after almost complete deceleration in the Bragg-Peak. Dose to normal tissue proximal and especially distal the treatment volume can be reduced. Therefore, proton beam therapy is of increasing interest in multimodality cancer care and gives the opportunity for improved local control by dose escalation or the reduction of treatment related toxicity. This is relevant especially for patients with tumors which are located in close proximity of critical structures or in particular sensitive tissues and indicate high treatment doses, like many sarcomas. For chordoma and chondrosarcoma of the skull base it was possible to show that by dose-escalated radiotherapy with protons local control could be improved with acceptable toxicity. Due to increasing treatment capacities proton therapy is becoming more widely available for this patient group. Treatment concepts for soft tissue sarcomas and bone sarcoma of the skull base, the spine, and the retroperitoneal region are under evaluation and are increasingly available for patient treatment. The West German Proton Therapy Center Essen (WPE) started treatments for patients in Mai 2013. Meanwhile more than 160 sarcoma patients were treated. 61% of the patients were children (age < 18 years). Current early prospective data suggest a good feasibility and tolerable side effects, while applying high treatment doses in difficult regions and sensitive normal tissue. Disclosure: No conflict of interest disclosed.

Fortbildung Melanom

Disclosure: No conflict of interest disclosed.

V327

V322

Circulating tumor DNA as biomarker for disease activity and response to treatment in BRAF V600E mutant malignant melanoma

Bone Sarcoma- therapeutic concepts Dirksen U.T.1 University Hospital Muenster, Paed Haem Onc, Muenster, Germany

Patients with newly diagnosed localized bone sarcoma are treated with a combination of multiagent cytotoxic chemotherapy and local control measures (surgery, radiation, or surgery with radiation) directed against the primary tumor. Cooperative group clinical trials such as EURAMOS-1, Euro E.W.I.N.G.- 99 and AEWS0031 provided evidence for the use of multidrug treatment. Results of the trials are presented and discussed.

Abstracts

von Bubnoff D.1, Follo M.2, Graf E.3, Pfeifer D.2, Duyster J.2, Meiß F.1, von Bubnoff N.2 Universitätsklinikum Freiburg, Klinik für Dermatologie und Venerologie, Freiburg, Germany, 2Universitätsklinikum Freiburg, Klinik für Innere Medizin I, Hämatologie, Onkologie und Stammzelltransplantation, Freiburg, Germany, 3 Institut für Medizinische Biometrie und Statistik, Freiburg, Germany 1

Introduction: Presence of BRAF V600E mutation in malignant melanoma tissue predicts for response to BRAF inhibitor treatment. However, this tumor specific mutation currently can not be used to monitor response to treatment or to predict relapse. Available biomarkers such

Oncol Res Treat 2016;39(suppl 3):1–104

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CONTENTS AUTHOR INDEX

as LDH or S100B exhibit limited sensitivity and specificity to predict response or progression in melanoma. We hypothesized that free plasma circulating tumor (ct)DNA containing BRAF V600E predicts outcome in BRAF V600E mutant melanoma. Methods: We developed a hydrolysis probe based, Locked Nucleic Acid assay to detect BRAF V600E and wild type ctDNA by droplet digital PCR on a BioRad QX100 system. Reaction sensitivity was 0.01% with high specificity. We analysed 107 plasma samples from 38 patients (pts) with stage III or stage IV BRAF V600E positive melanoma. The cohort included 30 unselected pts with 83 retrospective samples, 6 pts with baseline samples from a prospective trial (DRKS00009507), and 2 pts with 16 retrospective and 2 baseline samples from the prospective study. Results were correlated with LDH, S100 and imaging data. Results: Of 34 stage IV pts, 26 tested positive for BRAF V600E ctDNA at least once. Of 8 negative pts, 3 were in CR, 3 had stable disease, and 2 were negative baseline samples of the prospective trial. All 4 stage III pts were in CR, and 2 tested positive for BRAF V600E ctDNA at least once. Positive pts had a mean of 9 BRAF V600E copies/ml plasma (stage III; range: 1–17) and 483 copies/ml plasma (stage IV; range: 0,1–16.388). BRAF V600E ctDNA copies were strongly correlated with both LDH and S100 levels. Dynamic changes of BRAF V600E ctDNA correlated with disease course and response to treatment. A negative to positive conversion or increase was associated with progression (11 pts) or stable disease (1 pt) but not with response, whereas a positive to negative conversion or decrease of mutant BRAF copies correlated with response (10 pts) or mixed response (1 pt), but not with stable disease or progression. Conclusion: Dynamic changes in BRAF V600E ctDNA indicated response or progression in BRAF V600E mutated melanoma and correlated with LDH and S100. Based on these results, we have set up a prospective trial to determine the detection rate of BRAF V600E ctDNA in stage III and IV melanoma, and to analyze the predictive value of dynamic changes of BRAF ctDNA versus LDH and S100 with respect to clinical endpoints. Disclosure: Dagmar von Bubnoff: No conflict of interest disclosed. Nikolas von Bubnnoff: Financing of Scientific Research: Fa. Novartis, Fa. BMS; Expert Testimony: Fa. Novartis

Fortbildung

Blut ist ein besonderer Saft – Transfusionsmedizin V329

Choose wisely – Individualized and economic indications for platelet transfusions

3. Is a prophylactic transfusion strategy necessary for every patient with hypoproliferative thrombocytopenia? About 75% of all platelet products are used for hematological and oncological patients. For patients with hypoproliferative thrombocytopenia a prophylactic platelet transfusion strategy with a morning platelet trigger below 10/nl has been defined as standard practice by national and international guidelines. In recent years this practice has been challenged and a therapeutic platelet transfusion strategy has been tested in clinically stable patients. Two randomized studies compared these strategies. The prophylactic transfusion strategy has been shown to be more save in patients with acute leukemia. On the other side after autologous blood stem cell transplantation a therapeutic transfusion strategy could reduce the number of transfused units by about 30% and is not resulting in more severe bleeding. These questions will be discussed in detail. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium

Kontroversen in der Therapie des kolorektalen Karzinoms V335

Big data in the management of CRC. What is the benefit of extensive genomic analysis of the tumor? Wicki A.1,2 Universitätsspital Basel, Onkologie, Basel, Switzerland, 2Universität Basel, Dept. Biomedizin und Dept. Klinische Forschung, Basel, Switzerland 1

The term big data describes the gathering of structured and unstructured information, and the subsequent mining of large data silos. In cancer medicine, the largest data hub that has been tackled in the last few years is the tumor genome. Based on the genomic profile of more than 4000 patients with colorectal cancer, a new consensus classification of CRC has been proposed. In addition, the number of public institutions and private vendors that offer sequencing services is steeply increasing. In this presentation, I want to explore perspectives that are offered by extensive sequencing of colorectal cancer. What sampling is adequate? How do we deal with tumor heterogeneity? Are there patients who generally benefit from such an approach and to whom should we offer sequencing? Can we use sequencing to guide targeted and immune therapy? And finally, how can we best make use of sequence data in our daily practice? Disclosure: Andreas Wicki: Expert Testimony: Piqur Therapeutics, Basel, Switzerland; Actelion Ltd, Allschwil, Switzerland

Schäfer-Eckart K.1 Klinikum Nürnberg Nord, Medizinische Klinik 5, Nürnberg, Germany

About 500 000 platelet units are transfused in Germany every year. But platelet products are a limited resource and their use has to be judicious, especially as it has been assumed, that about 30–40% are not indicated according to national guidelines. To achieve not just a more economic but also an individual transfusion approach for patients with hypoproliferative thrombocytopenia three questions have to be answered. 1. What kind of platelet concentrate is appropriate for the individual patient? 60% of all platelet units used are single donor apheresis platelets, 40% are pooled platelet concentrates. While one platelet apheresis yields in 2 platelet concentrates, a pooled platelet concentrate can be produced with 4 whole blood donations. It has to be defined which patients really need an apheresis concentrate and for which the more economic and “donor sparing” pooled concentrate is appropriate. 2. What is the optimal platelet dose? A randomized study has shown, that the median platelet dose could be reduced to a dose below the national specification (1,1 x 1011 vs 3 x 1011) without a higher risk of severe bleeding.

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Fortbildung

Nierenzellkarzinom V336

Pathology and pathophysiology of renal cell carcinomas Haitel A.1 Institut für Klinische Pathologie der Medizinischen Universität Wien, AKH, Wien, Austria 1

The recently published WHO classification of Tumors of the Urinary System1 includes many different types of Renal Cell Carcinoma: • Clear cell renal cell carcinoma • Multilocular cystic renal neoplasm of low malignant potential • Papillary renal cell carcinoma • Hereditary leiomyomatosis and renal cell carcinoma-associated renal cell carcinoma (HLRCC) • Chromophobe renal carcinoma • Collecting duct carcinoma • Renal medullary carcinoma • MiT familily translocation renal cell carcinomas

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• Succinate dehydrogenase-deficient renal cell carcinoma • Mucinous tubular and spindle cell carcinoma • Tubulocystic renal cell carcinoma • Acquired cystic disease-associated renal cell carcinoma • Clear cell papillary renal cell carcinoma • Renal cell carcinoma, unclassified All these tumors show different morphology but also different immunohistochemical staining results. In addition they differ in prognosis and genetic profile. Clear cell renal cell carcinoma (ccRCC) shows VHL mutation in sporadic as well as hereditary ccRCC. This results in the loss of von Hippel-Lindau protein function which leads to progression and metastases. In addition they frequently show loss of heterocygosity (LOH)3p, which harbors VHLgene and at least four further tumor suppressor genes. The genetic profile of papillary renal cell carcinomas (pRCC) differs completely, as they show gains of Chromosome 7, 17 and loss of Y. In addition ca. 13% of sporadic pRCC and all pRCC of patients with hereditary pRCC show MET alteration. Sometimes pRCC show ALK (anaplastic lymphoma kinase) rearrangements. A distinct and very aggressive type of pRCC is that of HLRCC, associated with a germ line mutation of the fumarate hydratase gene located on Chormosome1. Chromosomes 1 and 17 are involved in translocation RCCs of the MiT family, too. TFE3 is located on Chromosom X and the two most common RCCs of this group are t(X;1)(p11.2;q21) which fuses PRCC and TFE3 genes and the t(X;17)(p11.2;q25) which fuses ASPL and TFE3, the same fusion found in alveolar soft part sarcomas. The latter are more likely to present with metastases.The translocation TFEB t(6;11) seems to be more indolent, only few cases are presented in the literature with metastases. Collecting Duct Carcinoma, also called Bellini duct carcinoma, is a very aggressive tumor. It shows LOHs on multiple chromosomes, but also trisomies and amplifications of Her2/neu have been reported. Reference: 1 WHO Class. of Tum. of the Urin. Syst. and Male Genital Org., Moch et al., Lyon, 2016. Disclosure: No conflict of interest disclosed. V337

S3 guideline for renal cell carcinoma – current treatment algorithm Grünwald V.1 MHH, Hannover, Germany

Introduction: The past decade of medical treatment in renal cell carcinoma (RCC) has been dominated by targeted therapies, which is a milestone in RCC treatment. An overall survival of 30 months remains the benchmark within clinical trials with the sequential use of the currently available therapies. The development of novel agents during the years have led to a more diverse environment and rendered the need for an update of the current S3 guideline in RCC. Methods: A systematic review for medical treatment of RCC was performed (from January 2013 on) and the evidence was graded according to SIGN criteria. Recommendation were given after consensus was achieved. Specifics on methodology can be found at: http://leitlinienprogramm-onkologie.de/Leitlinien.7.0.html. Results: The addition of cabozantinib and nivolumab to the armentarium has added value to the armentarium to treat RCC. Recommendations for their use within the context of the current landscape are given. Conclusions: After a fascinating decade of drug development in RCC, novel agents can be still added to the treatment algorithm.

Freier Vortrag

Immuntherapie solider Tumoren V339

Immune-related gene expression signatures as predictive biomarkers for outcome after concurrent chemoradiation in patients with locally advanced oropharyngeal carcinomas Heß A.-K.1, Joehrens K.2, Keilholz U.3, Rieke D.3, Weichert W.4, Balermpas P.5, Roedel C.5, Mairinger F.D.6, Hummel M.2, Augstein P.3,7, Budach V.1,7, Tinhofer I.1,7 Charité – Universitätsmedizin Berlin, Radioonkologie und Strahlentherapie, Berlin, Germany, 2Charité – Universitätsmedizin Berlin, Pathologie, Berlin, Germany, 3Charité Comprehensive Cancer Center, Berlin, Germany, 4Technische Universität München, Pathologie, München, Germany, 5DKFZ Heidelberg, Frankfurt, Germany, 6Universitätsklinikum Essen, Pathologie, Germany, 7DKFZ Heidelberg, Berlin, Germany 1

Introduction: The extent of immune-cell infiltration has previously been linked to the efficacy of concurrent chemoradiation (CRTX) in locally advanced squamous cell carcinoma of the head and neck. We established immune-related gene expression signatures in oropharyngeal carcinoma (OPC) patients from the ARO04–01 phase III trial as candidate predictive biomarker for cisplatin (CDDP)- and mitomycin C (MMC)-based CRTX. Potential overlap between the immune signatures of CRTX efficacy and the previously reported ‘inflamed’ signature of pembrolizumab activity was determined. Methods: Immune profiles were established from archival tumor specimens of OPC patients treated in the ARO04–01 study with concurrent MMC-CTRX (N = 24) or CDDP-CTRX (N = 23) were included. The CDDP-CTRX immune profile was validated in further 36 tumor specimens from the ARO04–01study (N = 7) and routine-care patients (N = 29). FFPE-extracted RNA was analyzed using the PanCancer Immune Profiling Panel on the NanoString nCounter® system. Gene expression signatures predictive for 3yr-OS were established by Spearman Rank correlation and hierarchical cluster analysis. Survival estimates according to the immune signatures were determined by Kaplan-Meier-analysis. The potential interference with the HPV status was evaluated in multivariate Cox regression models. Results: Signature 1 showed a strong positive association with OS (P < 0.001) and PFS (P = 0.006) after CDDP-CRTX but not MMC-CRTX. Its positive predictive value for 3yr-OS was 87.5%, and the negative predictive value was 100%; AUC = 0.97 [95% CI, 0.89–1.0]. Signature 1 was enriched for genes of the adaptive immune system, mainly affecting T-cell receptor and PD-1 signaling. A partial overlap with genes from the ‘inflamed’ pembrolizumab signature was observed. In contrast, signature 2 which showed a significant association with outcome for both CRTX regimens was enriched for genes of the innate immune system, regulating Toll-like receptor, TNF receptor and NFkappaB signaling. Despite a higher prevalence of signature 1 in HPV+ tumors, its predictive value for OS and PFS after CDDP-CRTX was independent of the HPV status. Conclusions: We demonstrate for the first time an impact of adaptive immunity and PD-1 signaling on the efficacy of CDDP-CRTX which may be important for inclusion of PD-1/PD-L1 inhibitors into curative treatment settings. The NanoString technology for immune gene expression analysis should provide a useful tool for patient stratification. Disclosure: No conflict of interest disclosed.

Disclosure: Viktor Grünwald: Employment or Leadership Position: keins; Advisory Role: BMS, Novartis, Pfizer; Financing of Scientific Research: BMS, Novartis, Pfizer

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PD-L1: a novel prognostic biomarker in head and neck squamous cell carcinoma Müller T.1, Brägelmann J.2, Dietrich D.1, Perner S.1, Kristiansen G.1, Bootz F.3, Brossart P.2 Universitätsklinikum Bonn, Institut für Pathologie, Bonn, Germany, Universitätsklinikum Bonn, Medizinische Klinik III, Bonn, Germany, 3 Universitätsklinikum Bonn, Klinik und Poliklinik für Hals-NasenOhrenheilkunde, Bonn, Germany 1 2

Background and Aims: PD-1/PD-L1 pathway seems to play a pivotal role in T cell downregulation during immune response to external antigens and self-antigens to prevent tissue damage and limit autoimmunity. Today there is no data available about PD-L1 expression in HNSCC, which induced us to analyze PD-L1 expression in HNSCC. Methods: Two independent cohorts of HNSCC (n1 = 98, n2 = 195) in a TMA format were analysed by immunohistochemistry (PD-L1, clone EPR1161 2 antibody) and evaluated by two observers. PD-L1 expression was scored semiquantitatively as negative vs. low vs. high. Statistical analysis was conducted with SPSS. Results: High expression levels of PD-L1 were detected in 15.3% of tumor cases in the training cohort and 27.7% of tumor specimens in the test cohort. Kaplan-Meier analysis demonstrated a significant prognostic value of PD-L1 in both cohorts (training cohort: p < 0.004; test cohort: p < 0.001), which was confirmed in a multivariate analyses encompassing recognized prognostic factors like tumor stage, nodal status, distant metastases, lymphatic invasion, vascular invasion , grading ,extracapsular expansion and surgical margin status (p = 0.02; HR = 2.926 [95%CI = 1.183 - 7.235]). Moreover strong PD-L1 expression was associated with the presence of distant metastases in the test cohort (p = 0.025). Conclusion: In summary we identified PD-L1 as expressed in HNSCC, which may provide a rational basis for anti PD-1/PD-L1 therapy in HNSCC patients. Additionally, it may serve as a multivariate prognostic biomarker in HNSCC, which deserves further study. Disclosure: Tim Müller: No conflict of interest disclosed. Peter Brossart: Advisory Role: BMS, MSD, Roche; Stock Ownership: Immatics; Financing of Scientific Research: BMS, MSD, Roche V341

Impact of nivolumab versus docetaxel on disease-related symptoms in patients with advanced non-squamous NSCLC from CheckMate 057 Horn M.1, Gralla R.J.2, Spigel D.R.3, Bennett B.4, Taylor F.4, Penrod J.R.5, DeRosa M.4, Dastani H.5, Strycker Orsini L.5, Reck M.1 Lungen Clinic Grosshansdorf, Airway Research Center North (ARCN), Grosshansdorf, Germany, 2Albert Einstein College of Medicine, Jacobi Medical Center, Bronx, United States, 3Sarah Cannon Research Institute, Medical Oncology, Nashville, United States, 4Adelphi Values, Boston, United States, 5 Bristol-Myers Squibb, Princeton, United States 1

Introduction: Nivolumab, a fully human IgG4 programmed death-1 immune checkpoint inhibitor, is approved in the US and EU for patients with previously treated advanced/metastatic NSCLC, based on results of 2 phase 3 trials: CheckMate 017 (NCT01642004) in squamous (SQ) NSCLC and CheckMate 057 (NCT01673867) in non-SQ (NSQ) NSCLC. We assessed the impact of nivolumab versus docetaxel on disease-related symptoms using the Lung Cancer Symptom Scale (LCSS) in patients with advanced non-SQ NSCLC from CheckMate 057. Methods: LCSS was evaluated every other cycle (Q4W) for nivolumab and every cycle (Q3W) for docetaxel for the first 6 mo on treatment, every 6 wk thereafter, and at 2 post-treatment follow-up visits. The LCSS has two components: the Average Symptom Burden Index (ASBI; based on 6 symptoms: anorexia, fatigue, cough, dyspnea, hemoptysis, and pain) and the 3-Item Global Index (3-IGI; symptom distress, interference with activities, and health-related quality of life [HRQoL]). Changes from baseline

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in LCSS scores and time to first deterioration (TTD) in symptoms were analyzed. Results: By wk 12, 17.8% (52/292) of nivolumab patients showed clinically meaningful symptom improvement (based on minimally important difference [MID]) compared with 19.7% (57/290) of docetaxel patients. Numerical differences in mean changes from baseline in the ASBI and 3-IGI favoring nivolumab appeared at the first common assessment (wk 12) and continued throughout the assessment period. At common assessments with >10 patients in each arm (wk 12–48), statistically significant differences favoring nivolumab were noted for the ASBI at wk 12, 24, 30, and 42, and for the 3-IGI at wk 24 and 30. At ≥1 common assessments, there were statistically significant differences favoring nivolumab for 5 of 6 symptoms (anorexia, fatigue, dyspnea, hemoptysis, and pain) and 2 of 3 items of the 3-IGI (HRQoL and symptom distress). Nivolumab versus docetaxel had a significantly longer TTD (based on MID) for the ASBI (HR = 0.65; 95% CI, 0.49–0.85) and 3-IGI (HR = 0.63; 95% CI, 0.48– 0.82), with Kaplan-Meier curves separating between arms at ≈2 months. A similar pattern occurred for TTD for individual ASBI symptoms and 3-IGI items. Conclusions: In CheckMate 057, patients with previously treated advanced non-SQ NSCLC had significantly lower symptom burden and better HRQoL while treated with nivolumab compared with docetaxel. Nivolumab- versus docetaxel-treated patients also demonstrated significantly longer TTD. Disclosure: Marlitt Horn: No conflict of interest disclosed. Martin Reck: Advisory Role: Consulting/advisory role: Roche, Lilly, BMS, MSD, Astra Zeneca, Pfizer, Boehringer-Ingelheim, Celgene; Other Financial Relationships: Speakers Bureau: Roche, Lilly, BMS, MSD, Astra Zeneca, Pfizer, Boehringer-Ingelheim, Celgene V342

Local treatment of tumors or metastases in combination with systemic ipilimumab immunotherapy prolongs overall survival in patients with advanced malignant melanoma Theurich S.1,2, Rothschild S.I.3, Hoffmann M.4, Fabri M.4, Sommer A.4, Garcia-Marquez M.2, Thelen M.2, Schill C.5, Merki R.5, Schmid T.6, Koeberle D.6, Zippelius A.3, Baues C.7, Mauch C.4, Tigges C.8, Kreuter A.8, Borggrefe J.9, Schlaak M.4, von Bergwelt-Baildon M.2 Uniklinik Köln, Klinik I für Innere Medizin, Köln, Germany, 2Uniklinik Köln, Interventionelle Immunologie Köln, Klinik I für Innere Medizin, Köln, Germany, 3 Universitätsspital Basel, Klinik für Innere Medizin, Onkologie, Basel, Switzerland, 4 Uniklinik Köln, Klinik für Dermatologie und Venerologie, Köln, Germany, 5 Kantonales Spital Aarau, Aarau, Switzerland, 6St. Claraspital Basel, Basel, Switzerland, 7Uniklinik Köln, Klinik für Strahlentherapie, Köln, Germany, 8HeliosKlinik St. Elisabeth, Klinik für Dermatologie und Venerologie, Oberhausen, Germany, 9Uniklinik Köln, Institut für Radiologie, Köln, Germany 1

Background: Immune checkpoint inhibition with ipilimumab has revolutionized cancer immunotherapy and significantly improved outcomes of patients with advanced malignant melanoma. Local peripheral treatments (LPT) such as radiotherapy or electrochemotherapy have been shown to modulate systemic immune responses. Preliminary data have raised the hypothesis that the combination of systemic immune checkpoint blockade with LPT could lead to improved clinical outcomes. Patients and methods: Clinical data of consecutively treated melanoma patients at four cancer centers in Germany and Switzerland were analyzed. Patients either received ipilimumab or ipilimumab and LPT if indicated for local tumor control. Additional immune assessments were performed in order to identify tumor-specific immune responses during the combination treatment. Results: A total of 127 melanoma patients were analyzed who either received ipilimumab (n = 82) or ipilimumab+LPT (n = 45). We found that the addition of LPT to ipilimumab significantly prolonged median overall survival (OS) (93 versus 42 weeks, p = 0.0028). As a potential immunological correlate, we identified significantly increased melanoma-specific T-cell responses following ipilimumab+LPT in one exemplary patient. Interestingly, adverse immune-related events were not significantly in-

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creased by the combination treatment. In a multivariable Cox regression analysis we demonstrate that the effect of added LPT on OS remained significant, after adjusting for BRAF status, tumor stage, tumor burden and CNS metastases (adjusted HR = 0.56, 95% CI = 0.31 to 1.01, p = 0.05). Conclusion: Our data suggest that the addition of LPT to ipilimumab is safe and effective in patients with metastatic melanoma irrespective of adverse disease characteristics. We hypothesize that enhancement of tumor-specific immune responses is most likely the underlying mechanism and that this combinatory approach warrants prospective validation. Disclosure: No conflict of interest disclosed. V343

Correlation of somatic mutational load as determined by panel based hybrid capture sequencing assay with tumor response to immunotherapy Griesinger F.1, Mariotti E.2, Menon R.2, Müller J.2, Lakis S.2, Walsh N.3, Grohe C.4, Crown J.3, Heuckmann J.M.2, Heukamp L.2 Universitätsklinik Innere Medizin-Onkologie, Klinik für Hämatologie und Onkologie, Oldenburg, Germany, 2NEO New Oncology, Cologne, Germany, 3 University College Dublin, Dublin, Ireland, 4Evangelische Lungenklinik Berlin, Berlin, Germany 1

Background: The use of immune checkpoint inhibitors in the field of cancer immunotherapy is revolutionizing cancer treatment. However, only a subset of tumors are responding, and powerful predictors for response are missing. Recent publications have emphasized on the importance of mutational load to identify patients who are likely to benefit from ‘mono’ or ‘combinational’ immunotherapy. Therefore, the use of a hybrid capture based next generation sequencing assay, like NEOplus, capable of reliably detected somatic mutational load in the diagnostic setting could support patient stratification for the treatment with immune checkpoint inhibitors. Method: Here, we describe an in silico analysis on the correlation of exonic territory and predictive power for response to immune checkpoint inhibitors based on published data. Furthermore, a broad range of tumor samples derived from multiple indications with corresponding patient response data were retrospectively analyzed for mutational load using a hybrid-capture based next-generation sequencing assay that covers clinically relevant genomic alterations in a panel of more than 90 genes, including point mutations, small insertions and deletions, selected gene fusions and copy number alterations. Results: We were able to show correlations between exonic territory covered by next-generation sequencing and predictive power for response to immune checkpoint inhibitors. In addition, panels covering only a selected sub-fraction of the human exome are able to reliably detect somatic mutational load in the analyzed tumor samples. Conclusion: Correlations between mutational load and patient response to immunotherapy were drawn in order to categorize patients who would most likely benefit from treatment.

V344

Efficacy, safety and predictive biomarker results from a randomized phase II study comparing atezolizumab vs docetaxel in patients with advanced NSCLC (POPLAR) Schulz C.1, Spira A.I.2, Park K.3, Mazieres J.4, Vansteenkiste J.5, Ballinger M.6, Waterkamp D.6, Fehrenbacher L.7 Universitätsklinikum Regensburg, Klinik und Poliklinik für Innere Medizin II, Bereich Pneumologie, Regensburg, Germany, 2Virginia Cancer Specialists Research Institute, Fairfax, United States, 3Samsung Medical Centre, Division of Hematology/Oncology, Seoul, Korea, Republic of, 4Toulouse University Hospital, Toulouse, France, 5University Hospitals KU Leuven, Leuven, Belgium, 6 Genentech, Inc., South San Francisco, United States, 7Kaiser Permanente Medical Center, Vallejo, United States 1

Introduction: In a Phase Ia clinical trial, atezolizumab (anti-PDL1) showed single-agent activity in NSCLC patients (pts) with the objective response rate (ORR) associated with PD-L1 expression on tumor-infiltrating immune cells (IC) and/or tumor cells (TC). Here we report results from the first randomized study comparing atezolizumab vs docetaxel (doc) in unselected NSCLC pts with subgroup analyses by PD-L1-expression. Methods: 287 previously-treated NSCLC pts were stratified by PD-L1 IC status, histology (squamous vs nonsquamous) and number of prior lines of therapy (1 or 2) and randomized (1:1) to 1200 mg IV q3w atezolizumab (n = 144) or 75 mg/m2 IV q3w doc (n = 143). PD-L1 expression was centrally evaluated using an immunohistochemistry (IHC) assay based on the SP142 antibody; pts were scored as IC 0, 1, 2 or 3 and TC 0, 1, 2, or 3. The objectives of this study were to evaluate the efficacy and safety of atezolizumab as compared with doc. The primary efficacy endpoint was overall survival (OS) and secondary endpoints were progression-free survival (PFS) and ORR. These endpoints will be assessed in both PD-L1-selected and the ITT patient populations. OS and PFS will be compared between treatment arms using the log-rank test; Kaplan-Meier methodology will be used to estimate median OS and PFS for each treatment arm and construct survival curves. Cox regression proportional hazard models will be used to estimate hazard ratios. An estimate of ORR (overall response of partial or complete response per RECIST v1.1) and its 95% CI will be calculated using the Clopper-Pearson method for each treatment arm. The incidence and severity of adverse events will be recorded. Enrollment was completed in March 2014. An analysis of the primary endpoint was performed in March 2015, with a minimum duration of follow-up of ≈ 11.5 mo. Results from the OS, PFS and ORR analyses for both the overall population and subgroups by IC and TC status will be presented. Demographic and safety data will be summarized. Disclosure: Christian Schulz: Advisory Role: Beratertätigkeiten für: AstraZeneca, Boehringer, BMS, Lilly, Novartis, Roche.; Financing of Scientific Research: Vortragshonorare von den Firmen: AstraZeneca, Boehringer, Celgene, MSD, Pfizer, Roche.; Other Financial Relationships: Reisekostenunterstützung von den Firmen AstraZeneca, Boehringer, BMS Louis Fehrenbacher: No conflict of interest disclosed.

Disclosure: Frank Griesinger: No conflict of interest disclosed. Lukas Heukamp: Employment or Leadership Position: NEO New Oncology; Stock Ownership: Stock ownership at NEO New Oncology

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Freier Vortrag

V346

Epigenetik V345

EZH2 mutations in myelodysplastic/myeloproliferative neoplasms Rinke J.1, Müller J.2, Bläß M.F.3, Chase A.4, Schäfer V.1, Winkelmann N.1, Haferlach C.5, Cross N.C.4, Hochhaus A.1, Ernst T.1 Abteilung Hämatologie und Internistische Onkologie, Klinik für Innere Medizin II, Universitätsklinikum Jena, Jena, Germany, 2Friedrich Schiller Universität Jena, Institut für Molekulare Zellbiologie, Jena, Germany, 3Klinik für Anästhesiologie und Intensivmedizin, Universitätsklinikum Jena, Jena, Germany, 4Wessex Regional Genetics Laboratory, Salisbury District Hospital, Salisbury, United Kingdom, 5MLL Münchner Leukämielabor, Munich, Germany 1

Introduction: Mutations in the epigenetic regulator gene EZH2 are frequently observed in myelodysplastic/myeloproliferative neoplasms (MDS/MPN; 10–13%). Here, we sought to genetically characterize a cohort of 40 EZH2-mutated MDS/MPN patients, to investigate mutational patterns associated with EZH2. Additionally, we analyzed clonal hierarchies of found mutations and evaluated how EZH2 mutations might influence genome-wide gene expression. Methods: The cohort included 40 MDS/MPN patients (male, n = 22; median age, 76 yrs; range, 61 to 88 yrs) with EZH2 mutations. Targeted deep next-generation sequencing (NGS) was used to analyze a panel of 30 commonly mutated genes in myeloid disorders. Clonal hierarchies were investigated in three patients using cryopreserved CD34+ cells to grow granulocyte-macrophage colonies which were analyzed using Sanger sequencing after DNA extraction and whole genome amplification. Stable shRNA-mediated down regulation of EZH2 in MOLM-13 was achieved via lentiviral transduction and confirmed by western blot analysis. Total RNA was used for genome wide expression analysis of cell lines to detect EZH2 targets, and compare whether these targets are differentially expressed in a subset of our cohort compared to EZH2-wildtype MDS/ MPN patients. Results: The NGS screen revealed a complex pattern of mutations with a total of 187 individual mutations. In addition to EZH2 mutations, 23 patients showed cooperating TET2 mutations (58%), followed by RUNX1 (40%), ASXL1 (33%), CBL (25%), ZRSR2 (20%), SRSF2 (15%), NRAS (15%) and STAG2 (15%). The majority of patients with mutations in TET2, RUNX1 and/or ASXL1 carried frameshift, nonsense or splice site mutations in these genes (83%, 81%, 92%, respectively). Colony assays showed a complex mutational hierarchy with various related subclones and the presence of a dominant clone. EZH2 mutations seemed to be an early event in leukemogenesis followed by SETBP1 mutations. Expression data is currently being analyzed. Conclusion: Here we have shown that EZH2 mutations are frequently associated with TET2, RUNX1 and ASXL1 mutations in MDS/MPN. EZH2 and U2AF35 mutations are mutually exclusive with fewer than expected SRSF2 mutations. Further unraveling mutational patterns and associations will help to better define disease entities and aid prognostic and therapeutic efforts. As EZH2 is an early event in leukemogenesis, it could be used for disease monitoring and may serve as a suitable drug target in the future.

Good response to epigenetic treatment with azacitidine in patients with myelodysplastic syndroms and chronic myelomonocytic leukemia is associated with clonal stabilization Speith J.1, Rinke J.1, Schäfer V.1, Gawlitza A.1, Waldau A.1, Hochhaus A.1, Ernst T.1 Abteilung Hämatologie und Internistische Onkologie, Klinik für Innere Medizin II, Universitätsklinikum Jena, Jena, Germany 1

Introduction: High-risk patients with MDS and CMML who do not qualify for an allogeneic stem cell transplantation can be treated with the demethylating substance azacitidine (Vidaza®). However, prediction of response to epigenetic therapy is difficult since the mechanism of action has not been elucidated sufficiently. Here, we aim to investigate the kinetics and evolution of molecular mutations in patients with MDS and CMML during treatment with azacitidine to get more insight into the molecular effect of this demethylating substance. Methods: We performed systematical mutational analysis of a clinically well-characterized cohort of patients with MDS-RAEB-I/II (n = 10) and CMML-I/II (n = 5) who responded well under treatment with azacitidine defined as a decrease of blasts < 5% in MDS or < 10% in CMML. Serial bone marrow samples at diagnosis and best clinical response were investigated by targeted deep next-generation sequencing (NGS) for a panel of 30 commonly mutated genes in myeloid disorders. Constitutional DNA obtained from buccal swabs was investigated to check somatic origin of mutations. Results: A total of 24 somatic mutations were identified in 8/10 MDS and in 5/5 CMML patients at diagnosis. Recurrently affected genes were TET2 (n = 5), RUNX1 (n = 4), EZH2 (n = 3), SRSF2 (n = 3), DNMT3A (n = 2), ASXL1 (n = 2), and KRAS (n = 2). Nearly 50% of patients (7/15) had ≥ 2 oncogenic mutations. A total of 23 somatic mutations were identified at the time of best response during treatment with azacitidine. Regarding mutation kinetics, 19/27 clones (70%) have already been observed at diagnosis and showed a complete stabilization of quantitative mutation load in 5 patients, 5/27 subclones (19%) decreased or disappeared (KRAS, n = 2; TP53, ASXL1, SRSF2) in five patients and 3 clones (11%) developed (KRAS, n = 2; ETV6) in three patients under epigenetic treatment. 3/5 patients with a clonal stabilization harbored TET2 mutations. Conclusion: In the majority of patients we were able to show that a response or remission of the disease under treatment with azacitidine was associated with a stabilization of the clonal architecture rather than a decrease or eradication of mutant subclones. Almost all patients with TET2 mutations achieved a clonal stabilization. Disclosure: No conflict of interest disclosed.

Disclosure: No conflict of interest disclosed.

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V348

Targeted sequencing reveals an accumulation of alterations in epigenetic regulators in relapsed ALL patients: data of the GMALL registry

The Allelic Ratio (AR) of DNMT3A R882 mutations (Mut) determined by digital droplet PCR (ddPCR) is a potential prognostic factor in Acute Myeloid Leukemia (AML)

Neumann M.1,2,3, Bastian L.1, Schlee C.1, Ortiz Tanchez J.1, Vosberg S.2,3,4, Rani James A.1,2,3, Schröder M.P.1, Schwartz S.1, Gökbuget N.5, Hoelzer D.5, Graf A.6, Krebs S.6, Blum H.6, Hecht J.7,8, Brüggemann M.9, Greif P.A.2,3,4, Baldus C.D.1,2,3

Grimm J.1, Dick T.1, Bill M.1, Jentzsch M.1, Schulz J.1, Schmalbrock L.1, Bonifacio L.1, Knyrim M.1, Schubert K.1, Cross M.1, Pönisch W.1, Vucinic V.1, Behre G.1, Franke G.-N.1, Niederwieser D.1, Schwind S.1

Charité – Universitätsmedizin Berlin (CBF), Hämatologie und Onkologie, Berlin, Germany, 2German Cancer Research Center (DKFZ), Heidelberg, Germany, 3German Cancer Consortium (DKTK), Heidelberg, Germany, 4LudwigMaximilians-University, Department of Internal Medicine 3, Munich, Germany, 5 Goethe University Hospital, Department of Medicine II, Hematology/Oncology, Frankfurt/M, Germany, 6Ludwig-Maximilians-University, Laboratory for Functional Genome Analysis, Gene-Center, Munich, Germany, 7The Barcelona Institute of Science and Technology, Centre for Genomic Regulation (CRG), Barcelona, Spain, 8Universitat Pompeu Fabra (UPF), Barcelona, Spain, 9University Hospital Kiel, Department of Hematology and Oncology, Kiel, Germany 1

Introduction: Targeted therapies based on molecular alterations are not established in standard treatment of ALL (with the exception of PhposALL). The example of Ph-like-ALL with proven efficacy of tyrosine kinase inhibitors underline the potential of identification of targetable lesions. Comprehensive analysis of mutational lesions in adult ALL is limited. Herein, we investigated the mutational spectrum in the routine diagnostic setting of the GMALL. Patients and methods: Over the period of 12 months, we prospectively investigated gDNA samples of 85 ALL patients (Phpos-BCP-ALL n = 23, Phneg-BCP-ALL n = 36, pro B-ALL n = 10, T-ALL n = 11, B-ALL n = 5). In addition, samples from 52 Phneg BCP-ALL patients were examined at first diagnosis and relapse. The GMALL gene panel comprised 206 genes with a target region of 1.05 Mbp (SureSelect, Agilent). Sequencing was performed on an Illumina HiSeq1500 (100bp paired-end) with 16 samples/ lane and an average read depth of ~600 reads/base. The turn-around-time was 4–6 weeks with incoming samples as the most limiting factor. Results: In Phneg-BCP-ALL (n = 98), the mutational spectrum was similar to childhood ALL, with the most alterations in KRAS (18%), PAX5 (16%), TP53 (9%), JAK1/2 (9%) and NRAS (8%). In T-ALL, NOTCH1 (6/11) and PHF6 (6/11) showed the most hits. Overall, 46% of all patients showed alterations in epigenetic modifiers. More than half of the patients (28/52, 54%) gained mutations in relapse, among those were known relapse related mutations like TP53 (n = 3) or CREBBP (n = 3). Ten patients (19%) acquired mutations in histone methylases or demethylases (KMT2D, SETD2, KMT2C, KDM6A, KDM5A). Notably, none of the 52 patients lost an existing mutation at first diagnosis in one of these genes. In one patient with a second relapse after blinatumumab therapy, we detected a FLT3 mutation in the tyrosine kinase domain. In experimental therapy with sorafenib, this patient showed a blast clearance in the peripheral blood and recovering of platelet count for a period of two months. Conclusions: The mutational screening using this GMALL gene panel at first diagnosis of an ALL is feasible. BCP-ALL shows a high rate of alterations in epigenetic modifiers at initial diagnosis and an increased accumulation in relapsed patients. Besides these, a number of detected alterations might offer therapeutic possibilities in relapsed or MRD-positive patients. It remains challenging to translate these findings into clinical practise.

Abteilung Hämatologie und Internistische Onkologie, Universitätsklinikum Leipzig, Leipzig, Germany 1

In AML the AR of various mut (e.g.FLT3-ITD) has been shown to impact on clinical outcome. However, little is known about DNMT3A R882mut in this respect. The highly sensitive & specific ddPCR may represent a feasible tool for AR assessment. AML patients (pts) treated at our institution between 2000 & 2015 with DNMT3A R882H or R882Cmut & diagnostic (n = 33) &/or follow-up (n = 20) cDNA samples were analyzed by ddPCR with a duplex assay measuring DNMT3Awildtype (wt) & mut. Absolute copy numbers of DNMT3Amut/DNMT3Awt defined the AR. Samples with an AR< 0.0001 or < 3 positive (+) droplets were considered negative (-) according to manufacturer’s recommendations. Mut in NPM1, CEPBA & presence of FLT3-ITD & -TKD were assessed. For pts receiving hematopoietic stem cell transplantation (HSCT; n = 27) data on bone marrow chimerism were collected. We identified 35 pts (median age 63 years [y], range 29–87y) with a DNMT3A R882H (71.4%) or R882Cmut (28.6%). All mut were heterozygous. At diagnosis pts with a DNMT3A R882mut AR>1 (31.4%, median AR 0.92) more often had secondary AML (P = 0.06), were less frequently NPM1mut (P = 0.03), had fewer platelets (P = 0.08), more blasts in peripheral blood (P = 0.04) & had shorter overall survival (P = 0.005, Figure1A). Of 15 pts with available cDNA up to 30 days before HSCT (pre-HSCT) 13 pts (86.7%) remained DNMT3A R882mut + & the AR declined in all pts (median AR 0.14, median AR reduction 67.3%, P < .001). 28 days after HSCT cDNA of 5 pts was available. One patient was DNMT3A R882mut - & is in continuous remission. 3 of 4 DNMT3A R882mut + pts (AR range 0.0001–0.51) experienced relapse after HSCT. Notably in 1 of those pts the DNMT3A R882mut AR increased by 2 log-levels between day 28 & 56 post-HSCT while the bone marrow total chimerism remained at 100% (Figure1B). Despite the limited number of pts our data suggest an association of the DNMT3A R882mut AR with biological features & prognosis in AML. We show that ddPCR is an eligible method to determine the DNMT3A R882mut AR & we aim to include the AR assessment in clinical trials to further investigate its prognostic influence & potential role as minimal residual disease marker after HSCT.

Disclosure: No conflict of interest disclosed.

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survival advantage of the NPM1mt AML xenograft and the Npm1CA/+RosaSB/+ model. Since MEIS1 was most dramatically suppressed, we assessed drug treatment effects on the MEIS1 target FLT3 that was also substantially downregulated. H3K79me2 levels across the HOX locus were also reduced upon MI503 treatment and pointed to DOT1L as having a role in HOX gene regulation. EPZ4777 treatment resulted in HOX, MEIS1, and FLT3 downregulation, cell growth inhibition, and profound differentiation of NPM1mt AML blasts. Next, we investigated effects of combined menin-MLL and DOT1L inhibition and found superior suppression of HOX and FLT3, superior differentiation effects, and synergistic growth inhibition. Combinatorial drug treatment further reduced leukemia initiating potential compared to single drug treatment in vivo. Conclusion: MLL and DOT1L are chromatin regulators that control leukemogenic gene expression in NPM1mt AML. Combinatorial small-molecule inhibition represents a therapeutic opportunity that should soon be ready for clinical assessment. Disclosure: Michael Kühn: No conflict of interest disclosed. Scott Armstrong: Advisory Role: Consultant for Epizyme, Inc. and Imago Biosciences. V350

Combined pharmacological DNMT and HDAC inhibition in AML cells: Integrative transcriptome and methylome analyses reveal synergistic gene downregulation associated with gene body demethylation

Fig. 1. Disclosure: No conflict of interest disclosed. V349

Synergistic inhibition of chromatin modifiers reverses a leukemogenic gene expression program in NPM1 mutant leukemia Kühn M.W.M.1,2, Song E.2, Feng Z.2, Sinha A.2, Chen C.-W.2, Deshpande A.J.2, Cusan M.2, Farnoud N.R.2, Koche R.P.2, Bradner J.E.3, De Stanchina E.4, Vassiliou G.S.5, Hoshii T.2, Armstrong S.A.2,6 Universitätsmedizin der Johannes Gutenberg-Universität Mainz, Medizinische Klinik III, Mainz, Germany, 2Memorial Sloan Kettering Cancer Center, Cancer Biology & Genetics Program, New York, United States, 3Dana Farber Cancer Institute, Harvard Medical School, Boston, United States, 4Memorial Sloan Kettering Cancer Center, Antitumor Assessment Facility, New York, United States, 5Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom, 6 Dana Farber Cancer Institute, Harvard Medical School, Pediatric Oncology, Boston, United States 1

Introduction: Aberrant HOX expression is commonly found in NPM1 mutant (NPM1mt) AML and FLT3 is concomitantly mutated in ~60% of these cases. Little is known how these cells maintain aberrant gene expression. Since MLL and DOT1L control HOX expression in other settings, we investigated potential dependencies of NPM1mt AML on these chromatin modifiers and delineated their potential role in regulating leukemogenic gene expression. Methods: CRISPR-Cas9 genome editing across exons encoding specific MLL protein domains was used in a negative selection screen to assess dependencies of NPM1mt AML. This concept was extended with inhibitors of the menin-MLL interaction (MI2–2; MI503). Dependencies on DOT1L were explored using the inhibitor EPZ4777. Target validation was performed in a human xenograft and two conditional murine knock-in models of Npm1mt AML (Npm1CA/+Flt3ITD/+; Npm1CA/+RosaSB/+). Results: CRISPR-Cas9 negative selection screening revealed NPM1mt OCI-AML3 cells to be exceptionally dependent on the menin binding site in MLL. Pharmacological menin-MLL inhibition resulted in dramatic HOX and MEIS1 suppression, differentiation induction, and profound growth inhibition in human OCI-AML3 and murine Npm1mt AML cells. Of interest, ectopic expression of Meis1, Hoxb4, or Hoxa9-Meis1 rescued the antiproliferative drug effects. In vivo MI503 treatment resulted in HOX and MEIS1 suppression, reduction of leukemia burden, and a significant

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Schlosser P.1,2, Blagitko-Dorfs N.2, Greve G.2,3, Pfeifer D.2, Lübbert M.2 Medical Center – University of Freiburg, Institute for Medical Biometry and Statistics, Freiburg, Germany, 2Medical Center – University of Freiburg, Department of Hematology, Oncology and Stem Cell Transplantation, Freiburg, Germany, 3Faculty of Biology, University of Freiburg, Freiburg, Germany 1

Background: Transcriptome-wide studies have shown that HMAs can downregulate mRNAs. Recently, Yang et al.showed that gene body methylation can be strikingly reversed by DAC in cancer cells, with downregulation of genes regulated by c-MYC, implicating gene body methylation as a novel drug target. Therefore, we asked on a transcriptome- and methylome-wide level whether HMA-induced reversal of gene body methylation occurs in AML, and whether this is associated with gene repression. Materials and methods: U937 cells were treated with 3×24h pulses of DAC (50nM), alone or in combination with the HDACi panobinostat (Pano, 8nM) or Valproic acid (VPA, 1mM) for 24h. Transcriptome and methylome profiles (biological triplicates) were generated using Affymetrix Human Gene 2.0 ST and Infinium HumanMethylation450 BeadChip arrays. Data were analyzed using R package RnBeads applying standard methods. Results: Transcriptome analyses revealed that DAC, Pano or VPA alone induced 207, 267 and 179 transcripts, with downregulation of 28, 45 and 22 transcripts, respectively (FDR< 0.01; ≥2-fold change). When DAC was combined with either Pano (D+P) or VPA (D+V), a striking synergistic effect was observed for upregulated transcripts. Remarkably, a synergistic effect on downregulated transcripts was even stronger. As expected, DAC treatment resulted in extensive DNA demethylation: 295.6K CpGs became >10% demethylated (FDR< 0.01) The ratio of transcripts mapping to promoter- vs. gene body-demethylated CpGs doubled from up- to downregulated transcripts (all 3 treatments). Of the 667 transcripts downregulated by D+P, the 227 gene-body demethylation-associated transcripts included various putative or bona fide oncogenes (such as c-MYC) and, notably, many epigenetic modifiers. Gene ontology analysis revealed an enrichment for genes involved in RNA processing and transcription validated for several genes encoding histone methylation “writer” enzymes by RT-qPCR. Conclusions: The combination of a HMA with either of two HDACi resulted not only in synergistic gene reactivation but also in massive, synergistic downregulation of genes including oncogenes and histone modifying enzymes, which was associated predominantly with gene body DNA demethylation. These findings have implications for the mechanisms of

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action of combined epigenetic treatment and for a better understanding of clinical responses in trials where this approach is tested, such as the randomized DECIDER AML trial. Disclosure: Pascal Schlosser: No conflict of interest disclosed. Michael Lübbert: Financing of Scientific Research: Novartis, Janssen-Cilag; Expert Testimony: Janssen-Cilag

Expertenseminar

Patienten mit ZNS-Lymphomen

Freier Vortrag

Akute myeloische Leukämie – Therapie 2 V353

The new WHO classification (2016) defines a revised diagnostic approach to myeloid neoplasms with erythroid precursors comprising ≥ 50% of bone marrow nucleated cells: Applicability and clinical impact Haferlach T.1, Kern W.1, Perglerova K.2, Haferlach C.1 MLL Münchner Leukämielabor GmbH, München, Germany, 2MLL 2, Paříkova, Czech Republic 1

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Primary central nervous system lymphoma in the elderly Illerhaus G.

Klinikum Stuttgart, Hämatologie, Onkologie und Palliativmedizin; Stuttgart Cancer Center, Stuttgart, Germany 1

Patients older than 60 years account for 50% of all PCNSL cases. Although elderly patients are able to tolerate aggressive systemic chemotherapy, they have an inferior prognosis compared with younger patients and are more seriously affected by iatrogenic toxicity, especially neurological side-effects following whole brain radiotherapy. Therefore, elderly patients represent a unique and vulnerable treatment subgroup. An US registry study of 579 elderly patients diagnosed with PCNSL in the 1990s revealed that the median survival was only 7 months and WBRT alone was the most common treatment modality (46%). The addition of rituximab to highdose methotrexate (HD-MTX)-based chemotherapy has shown a significant survival improvement in the IELSG-trial prospective, however, this trial was not particularly designed for elderly patients. Based on a systematic review of studies focusing on elderly PCNSL patients, there is only one completed randomized phase II trial for elderly patients, which suggests that a combination of HD-MTX with vincristine, procarbazine, and cytarabine is more effective than HD-MTX with temozolomide regarding response; however, difference in progression free survival were only marginal and no rituximab was used in this study. Data from the recently completed multicenter German PRIMAIN study strengthen the evidence that immunochemotherapy with HD-MTX combined with an oral alkylating agent (e.g. procarbazine) together with rituximab is a feasible and effective treatment and could provide a benchmark for future comparative studies. Still, treatment-associated morbidities and treatment delays are major issues that need to be addressed in future trials e.g. by increasing rituximab density in the beginning and introduction of better tolerable novel agents. To reduce the risk of relapse, immuno-modulatory agents (e.g. lenalodomide) could be attractive options to be tested as maintenance treatment. High-dose chemotherapy followed by autologous stem cell transplantation (HDT-ASCT) has been shown to be feasible and effective in younger patients. A pilot study investigating HDT-ASCT feasibility in elderly patients is currently ongoing. In summary, HD-MTX with oral alkylating agents (e.g. procarbazine or temozolomide) and rituximab are advisable and should be offered to elderly and frail patients. The role of HDT-ASCT in fit elderly patients is under investigation. More prospective trials designed for elderly PCNSL patients are needed.

Introduction: The 2016 revision to WHO classification of myeloid neoplasms (Arber et al., BLOOD, prepub 11.04.2016) revises AML with ≥ 50% bone marrow erythroid precursors and ≥20% myeloid blasts among non-erythroid cells. In the new classification the denominator for determining the percentage of myeloid blasts is total bone marrow cells. Thereby, erythroid leukemias showing blast cells ‹20% of the total cells are now classified as MDS. Further, cases with NPM1mut, CEBPAdmut, RUNX1mut all qualify for genetically defined entities. Patients and methods: We here investigated 190 cases of AEL (WHO 2008/FAB AML M6) all diagnosed in our laboratory between 2005–2016. Results: Following WHO 2016 we first subtracted cases with NPM1 (n = 28), CEBPAdm (n = 2) and RUNX1 (n = 16) mutations (46/190, 24%). We then separated cases according to BM blasts and found 35% (51/144) cases with ≥20% total blasts. 18/51 (35%) classify as “myelodysplasia-related changes” due to their cytogenetics and/or multilineage dysplasia. The remaining 33 (65%) cases with ≥20% of total blasts now are AEL. The remaining 93/144 (65%) cases showed < 20% total blasts and are now MDS. Within these 48/93 cases (52%) show “myelodysplasia-related changes”. We now compare new AEL (n = 33) and new MDS (n = 45). Both new subgroups show a normal karyotype in 73% each. In AEL and MDS the following mutations were observed at not significantly different frequencies: MLL-PTD (39%, 26%), ASXL1 (18%, 16%), FLT3-ITD (17%, 4%), TET2 (0%, 20%), SF3B1 (0%, 13%). Only IDH1 showed a statistically different distribution (AEL 27%, MDS 0%, p = 0.046). Using univariate Cox regression analysis, in all 144 patients only cytogenetics classified as “myelodysplasia-related changes” (p = 0.013; HR 3.1) and TP53 mutation (p = 0.001; HR 6.4) showed significant negative prognostic impact. We did not find any effect for BM blasts %. In multivariate analysis only mutated TP53 showed a significant and poor outcome (p = 0.01 HR 7.5). Regarding survival we did not find significant differences between AEL (n = 15; 2-yrs OS: 64%) and new MDS cases (n = 24; 2-yrs OS: 66%). Conclusions: AEL/AML M6 were newly defined in WHO 2016. Based on 190 patients according to WHO 2008, we find that 35% of patients remain AML and 65% of patients are MDS now. Genetics and outcome seem to be quite overlapping. Patients with the respective morphological features should be treated according to genetics rather than number of myeloid blasts in the BM. Disclosure: Torsten Haferlach: Employment or Leadership Position: Geschäftsführer MLL Münchner Leukämie Labor Claudia Haferlach: Employment or Leadership Position: Geschäftsführerin MLL Münchner Leukämie Labor

Disclosure: Gerald Illerhaus: Financing of Scientific Research: Riemser; Expert Testimony: Riemser

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V355

A Fc-optimized FLT3 antibody for elimination of minimal residual disease (MRD) in patients with AML

Long-term outcome of sorafenib treatment in FLT3-ITDpositive acute myeloid leukemia patients relapsing after allogeneic stem cell transplantation

Dörfel D.1,2, Körner S.1,2, Nübling T.1,2, Bamberg M.3, Bühring H.-J.1, Hofmann M.3, Grosse-Hovest L.4, Aulwurm S.4, Bethge W.1, Kanz L.1, Jung G.3, Salih H.R.1,2 Department of Hematology and Oncology, Eberhard Karls University, Tübingen, Germany, 2Clinical Collaboration Unit Translational Immunology, German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Partner site Tübingen, Tübingen, Germany, 3Department of Immunology, Eberhard Karls University, Tübingen, Germany, 4Synimmune GmbH, Tübingen, Germany 1

In the majority of patients with AML, a morphological complete remission (CR) can be achieved by chemotherapeutic treatment. However, minimal residual disease (MRD) remains detectable in many patients. Recently, persistance of MRD defined e.g. by detection of NPM1-mutated transcripts (mutNPM1) was identified as powerful prognostic parameter for relapse (e.g., Ivey et al, NEJM 2016). Here we report our clinical observations upon application of a novel Fc-optimized antibody (4G8-SDIEM, FLYSYN) directed to the receptor tyrosine kinase FLT3 that is expressed on the surface of AML cells. Our antibody was developed with the aim to eliminate MRD after conventional therapy and to achieve this goal with less toxicity (i.e. GvHD) than donor lymphocyte infusion (DLI) and allogeneic stem cell transplantation (SCT). After preclinical characterization, which revealed its profound ability to mediate ADCC against AML cells (Hofmann et al, Leukemia 2012), the antibody was produced in a university-owned GMP-unit and applied on a compassionate need basis to 8 AML patients (male, 4; female, 4; median age, 60 years, (range 42–76)) in CR with persistant or increasing MRD (mutNPM1, n = 5; CBFB-MYH11 n = 1, AML-ETO1 n = 1, KITD816V n = 1). 5 patients had undergone SCT, 3 had refused or were unfit for SCT. FLT3 surface expression on leukemic blasts was confirmed by FACS in 6/8 patients. The antibody was applied in a dose of 25 mg total, and patients were then followed for clinical status and MRD levels. No relevant side effects (in particular no hematological toxicity) were observed. The serum half-live was found to range from 39–166 hours. In 3 patients (notably all with mutNPM1 as MRD marker), the single application of 4G8SDIEM (in 2 cases subsequently followed by treatment with one dose of DLI) resulted in rapid elimination of MRD. One patient that had received DLI died of complications due to GvHD and liver toxicity after reaching and maintaining a molecular CR. The other two patients (1 with, 1w/o DLI) remain in molecular CR for at present 44 and 42 months after 4G8-SDIEM application without further specific treatment. These results imply that 4G8SDIEM might be capable to reduce or eliminate MRD after conventional therapy at the cost of very few side effects. Based on these observations, a phase I/II trial enrolling AML patients ineligible for SCT in CR with detectable mutNPM1 is presently submitted to the regulatory authorities and envisaged to start recruitment in July 2016. Disclosure: No conflict of interest disclosed.

Metzelder S.K.1, Schroeder T.2, Lübbert M.3, Götze K.4, Scholl S.5, Ditschkowski M.6, Meyer R.G.7, Dreger P.8, Basara N.9, Fey M.10, Salih H.11, Finck A.3, Finke J.3, Pabst T.10, Giagounidis A.12, Neubauer A.1, Burchert A.1 Department of Hematology/Oncology/Immunology, University Hospital Marburg, Marburg, Germany, 2University of Duesseldorf, Medical Faculty, Department of Hematology, Oncology and Clinical Immunology, Düsseldorf, Germany, 3University of Freiburg, Department of Hematology/Oncology, Freiburg, Germany, 4Dept. of Internal Medicine III, Technische Universität München, München, Germany, 5Abteilung Hämatologie/Onkologie, Universitätsklinikum Jena, Jena, Germany, 6Department of Bone Marrow Transplantation, West German Cancer Center, University Hospital Essen, Essen, Germany, 7St.-Johannes-Hospital Dortmund, Klinik für Innere Medizin II, Dortmund, Germany, 8Department of Medicine, University of Heidelberg, Heidelberg, Germany, 9Malteser Krankenhaus St. Franziskus-Hospital, Medizinische Klinik I, Flensburg, Germany, 10Department of Medical Oncology, Inselspital and University of Bern, Bern, Switzerland, 11Department of Hematology/Oncology, Eberhard Karls-University, Tübingen, Germany, 12Marien Hospital Düsseldorf, Klinik für Onkologie, Hämatologie und Palliativmedizin, Düsseldorf, Germany 1

In acute myeloid leukemia (AML) the presence of an internal tandem duplication (ITD) in the FLT3 gene predicts a poor prognosis. We recently reported on 65 FLT3-ITD positive AML patients, who were given sorafenib monotherapy for relapsed or refractory disease, including 29 patients, who relapsed after allogeneic stem cell transplantation (allo-SCT) (Metzelder et al. 2012). The long-term efficacy of sorafenib monotherapy in this situation of ultra-poor risk AML is ill defined. Here we report on the long-term follow-up of these 29 patients. The median follow up in this cohort was 6.4 years (range, 4–8.6). Six of the 29 patients (21%) are still alive. Excluding one patient who received a 2nd allo-SCT after developing sorafenib resistance, five of the 29 patients experience long-lasting remissions with a 5-year overall survival of 17%. After a median treatment duration of 29 months (range, 18–39) sorafenib treatment was stopped in four patients. One of these patients relapsed eleven months after stopping sorafenib but is currently in complete molecular remission 56 months after restarting sorafenib. The other three patients are still in treatment free remission for 31, 37 and 59 months. For the first time we show not only that sorafenib monotherapy can induce durable remissions in a selected group of relapsed ITD positive AML patients after allo-SCT but also that stopping sorafenib after prolonged treatment is possible with a realistic chance of curation. Disclosure: No conflict of interest disclosed. V356

Persistence of driver mutations during complete remission associates with shorter survival and contributes to the inferior outcomes of elderly patients with acute myeloid leukemia Rothenberg-Thurley M.1, Amler S.2, Görlich D.2, Sauerland M.C.2, Schneider S.1, Konstandin N.P.1, Schaaf S.3, Nazeer Batcha A.M.3, Bräundl K.1, Ksienzyk B.1, Zellmaier E.1, Mansmann U.3,4, Fiegl M.1, Subklewe M.1, Bohlander S.K.5, Faldum A.2, Hiddemann W.1,4, Spiekermann K.1,4, Braess J.6, Metzeler K.H.1,4, AMLCG Medizinische Klinik III, Klinikum der Universität München, München, Germany, Institut für Biometrie und Klinische Forschung, Universität Münster, Münster, Germany, 3Institut für medizinische Informationsverarbeitung, Biometrie und Epidemiologie, LMU München, München, Germany, 4Deutsches Konsortium für Translationale Krebsforschung (DKTK), Heidelberg, Germany, 5Department of Molecular Medicine and Pathology, University of Auckland, Auckland, New Zealand, 6Klinik für Onkologie und Hämatologie, Krankenhaus Barmherzige Brüder, Regensburg, Germany 1 2

Introduction: AML patients (pts) in complete morphological remission may harbor persistent preleukemic clones, which might be a source of leukemic relapse. Previous studies demonstrated an adverse outcome of

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pts with persistent mutations in remission (Klco et al., JAMA 2015; PMID: 26305651). We studied persistence of driver mutations in remission and outcomes in a cohort of uniformly treated AML patients. Patients and methods: We studied 108 adult AML pts (median age, 56 years [y]; range 20–80y) who received intensive induction chemotherapy in the multicenter AMLCG-2008 trial, and reached either complete remission (CR) (74%) or CR with incomplete blood count recovery (CRi) (26%). In bone marrow (BM) or blood (pB) specimens obtained at diagnosis (BM, n = 96; pB, n = 12) and during first remission (BM, n = 102; pB, n = 6), we analyzed 68 recurrently mutated genes by multiplexed amplicon sequencing. Results: At diagnosis, 426 mutations were detected in 42 genes (median, 4 mutations per patient; range, 0–10). In the paired remission samples, 61 mutations were still present in 38/107 pts (36%), while 69 pts (64%) had no persistent mutation. Persistence of mutations during morphological remission was most commonly observed for DNMT3A (23/37 pts with mutations at diagnosis), TET2 (8/13), SRSF2 (5/8) and ASXL1 (5/12). Mutations in other genes including NPM1, FLT3, WT1, and NRAS were cleared in the remission samples. Pts with ≥1 persisting mutations in remission were older (median, 63y) than pts without any persisting mutation (median, 48y; p < 0.001). Persistence of ≥1 mutation in remission, in contrast to complete mutation clearance, associated with shorter relapse-free survival (RFS; Fig. 1A) and overall survival (OS; Fig. 1B). In multivariate analyses adjusting for age, ELN genetic risk groups, and remission status (CR vs. CRi), detection of ≥1 persisting mutation remained associated with inferior RFS (HR, 2.1; P = 0.02) and OS (HR, 2.5; P = 0.02). Conclusion: Detection of persisting mutations during first CR/CRi is common in older AML pts. Mutation persistence associates with shorter RFS and OS and might contribute to the inferior outcomes of elderly AML pts.

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BCR-ABL+ acute myeloid leukemia: a rare and intriguing entity – a retrospective analysis of clinical and molecular features Neuendorff N.R.1, Burmeister T.1,2, Dörken B.1,2, Westermann J.1,2 Charité Universitätsmedizin Berlin, Klinik für Hämatologie, Onkologie und Tumorimmunologie, Campus Virchow Klinikum, Berlin, Germany, 2Labor Berlin Charité-Vivantes GmbH, Berlin, Germany 1

Introduction: BCR-ABL+ AML is a rare subgroup of AML, likely to be included in the new WHO classification as a provisional entity. The distinction between BCR-ABL+ AML and CML blast crisis (CML-BC) is often challenging. We analysed all cases published since 1975 and from our institution regarding clinical and molecular features for a better characterization and distinction from CML-BC. Methods: BCR-ABL+ AML cases were obtained by a Pubmed literature search and from our institutional database. For comparability, cases were (if necessary) re-classified according to the latest WHO classification or, in the case of AUL/MPAL, according to EGIL and/or WHO classification. AUL/MPAL cases were excluded from the analysis, likewise those with an antecedent myeloproliferative disorder and cases with 100% BCR-ABL+ metaphases (published before 1990), if no additional features were given to distinguish AML from CML-BC. Finally, 126 cases of de novo BCRABL+ AML were analysed. Results: According to WHO 2008, 38% of the cases can be categorized as “AML-NOS”, 32.5% as “AML with myelodysplasia-related changes” and 23.8% as “AML with recurrent genetic aberrations” (mostly CBF leukemia). In a substantial portion, co-expression of lymphoid markers was present, not fulfilling the criteria for classification as “MPAL/AUL”. Basophilia was seen in only 5.6% of cases, splenomegaly in 16.7%. BCRABL transcripts (p190 and p210) were equally distributed. According to ELN criteria (irrespective of BCR-ABL), 21.4% belonged to the “favorable”, 39.7% to the “intermediate-I/II” and 30.2% to the “adverse” risk group (8.7% could not be classified due to a lack of data). Remarkably, long-term survival without allogeneic stem cell transplantation was reported within the favorable risk group. Conclusions: To the best of our knowledge, our study provides the most comprehensive overview on BCR-ABL+ AML published so far. With regard to our database with 126 de novo BCR-ABL+ AML we have developed a diagnostic algorithm including patients´medical history, basophilia, splenomegaly, the type of BCR-ABL transcript, the percentage of BCR-ABL+ metaphases in the bone marrow and cytogenetics. Furthermore, our data suggest the prognosis of BCR-ABL+ AML is rather determined by the specific genetic background than by BCR-ABL itself. Finally, we conclude that treatment with tyrosine kinase inhibitors remains an individual decision, which may be particularly appropriate beyond first line therapy. Disclosure: Nina Neuendorff: Other Financial Relationships: Stipendium der Firma Medac Jörg Westermann: Advisory Role: Novartis, BMS, Celgene; Financing of Scientific Research: Novartis, BMS, Celgene; Expert Testimony: Novartis

Fig. 1. Disclosure: No conflict of interest disclosed.

Abstracts

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V358

Real life experience with ATRA-arsenic trioxide based regimen in acute promyelocytic leukemia – results of the prospective German intergroup Napoleon registry Platzbecker U.1, Lengfelder E.2, Götze K.3, Röllig C.1, Kramer M.1, Sauer M.1, Albers N.4, Heuser M.5, Link H.6, Schäfer-Eckart K.7, Martin S.8, Krause S.9, Hänel M.10, Bochtler T.11, Jakob A.12, Reichle A.13, Biersack H.14, Baldus C.15, Görner M.16, Basara N.17, Mezger J.18, Naumann R.19, Kürschner D.20, de Wit M.21, Nolte F.22, Scholl S.23, Ringhoffer M.24, Tischler H.J.25, Greiner J.26, Kremers S.27, Machherndl-Spandl S.28, Koller E.29, Wattad M.30, Aschauer G.31, Radsak M.32, Kubin T.33, Salih H.34, Wolf D.35, Thiede C.1, Serve H.36, Dührsen U.37, Hochhaus A.23, Brümmendorf T.38, Bornhäuser M.1, Döhner H.39, Ehninger G.1, Schlenk R.F.39 Universitätsklinikum Carl Gustav Carus, Medizinische Klinik und Poliklinik I, Dresden, Germany, 2Universitätsklinikum Mannheim, III. Medizinische Klinik, Mannheim, Germany, 3Klinikum rechts der Isar der Technischen Universität München, III. Medizinische Klinik, München, Germany, 4Gesellschaft für Medizinische Innovation – Hämatologie und Onkologie mbH (GMIHO), Berlin, Germany, 5Medizinische Hochschule Hannover, Klinik für Hämatologie, Hämostaseologie, Onkologie und Stammzelltransplantation, Hannover, Germany, 6Westpfalz-Klinikum GmbH, Medizinische Klinik I, Kaiserslautern, Germany, 7Klinikum Nord, Medizinische Klinik V, Nürnberg, Germany, 8 Robert-Bosch-Krankenhaus Stuttgart, Abteilung Hämatologie, Onkologie und Palliativmedizin, Stuttgart, Germany, 9Universitätsklinikum Erlangen, Medizinische Klinik V, Erlangen, Germany, 10Klinikum Chemnitz, Klinik für Innere Medizin III, Chemnitz, Germany, 11Universitätsklinikum Heidelberg, Medizinische Klinik V, Heidelberg, Germany, 12Ortenau Klinikum, Offenburg, Germany, 13Universitätsklinikum Regensburg, Klinik für Hämatologie und Internistische Onkologie, Regensburg, Germany, 14Universitätsklinikum Lübeck, Medizinische Klinik I, Lübeck, Germany, 15Charité Berlin, Campus Benjamin Franklin, Medizinische Klinik mit Schwerpunkt Hämatologie, Onkologie, Berlin, Germany, 16Klinikum Bielefeld Mitte, Klinik für Hämatologie, Onkologie und Palliativmedizin, Bielefeld, Germany, 17Malteser Krankenhaus St. FranziskusHospital, Medizinische Klinik I, Flensburg, Germany, 18St. Vincentius Kliniken, Medizinische Klinik II, Karlsruhe, Germany, 19Gemeinschaftsklinikum Mittelrhein, Klinik für Innere Medizin, Koblenz, Germany, 20MVZ Mitte und MVZ Delitzsch, Delitzsch, Germany, 21Vivantes Klinikum Neukölln, Klinik für Hämatologie und Onkologie, Berlin, Germany, 22St. Hedwig-Krankenhaus, Klinik für Innere Medizin, Berlin, Germany, 23Universitätsklinikum Jena, Klinik für Innere Medizin II, Jena, Germany, 24Städtisches Klinikum Karlsruhe, Medizinische Klinik III, Karlsruhe, Germany, 25Johannes Wesling Klinikum Minden, Zentrum für Innere Medizin, Minden, Germany, 26Diakonie-Klinikum Stuttgart, Medizinische Klinik II, Stuttgart, Germany, 27Caritas-Krankenhaus Lebach, Innere Medizin, Lebach, Germany, 28Krankenhaus der Elisabethinen Linz, Interne Abteilung, Linz, Austria, 29Hanuschkrankenhaus Wien, III. Medizinische Abteilung, Wien, Austria, 30 Kliniken Essen Süd, Ev. Krankenhaus Essen-Werden, Zentrum für Innere Medizin – Hämatologie, internistische Onkologie u. Stammzelltransplantation, Essen, Germany, 31Krankenhaus der Barmherzigen Schwestern, Abteilung für Innere Medizin I, Linz, Austria, 32Klinikum der Johannes Gutenberg Universität Mainz, III. Medizinische Klinik und Poliklinik, Mainz, Germany, 33Klinikum Traunstein, Innere Medizin, Abteilung Hämato-Onkologie, Traunstein, Germany, 34 Universitätsklinikum Tübingen, Department für Innere Medizin, Innere Medizin II, Tübingen, Germany, 35Universitätsklinikum Bonn, Medizinische Klinik und Poliklinik III, Bonn, Germany, 36Universitätsklinikum der J.W. Goethe-Universität Frankfurt, Medizinische Klinik II, Frankfurt, Germany, 37 Universitätsklinikum Essen, Klinik für Hämatologie, Essen, Germany, 38 Universitätsklinikum Aachen, Medizinische Klinik IV, Aachen, Germany, 39 Universitätsklinikum Ulm, Innere Medizin III, Ulm, Germany 1

Standard therapy of acute promyelocytic leukemia has long time relied on the combination of All-trans-retinoic acid (ATRA) and chemotherapy. The introduction of arsenic trioxide (ATO) in APL treatment has allowed obtaining similarly high remission and even superior survival rates coupled with significantly reduced myelosuppression. Recent results of the APL0406 trial by the GIMEMA-AMLSG-SAL study groups showed that the combination of All-trans retinoic acid (ATRA) and arsenic trioxide (ATO) is superior to standard ATRA and chemotherapy (CHT) in frontline therapy of low/intermediate risk acute promyelocytic leukemia (APL). The translation of these results into clinical practice for APL patients in Germany are uncertain also given the fact that ATO is not approved by the European Medical Agency for front-line therapy in APL. In order to provide evidence and a reflection of the clinical reality of APL patient care

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in Germany an intergroup APL registry (NAPOLEON) has been initiated recently by German AML study groups. Eligibility is defined as age of at least 18 years and newly diagnosed or relapsed APL. Here we report the first series of patients prospectively enrolled into this registry. The study is conducted in accordance with the Declaration of Helsinki, received IRB approval by all participating centers and is registered at ClinicalTrials.gov (NCT02192619). As of 1st of April 2016 a total of 84 patients have been included into the study with a median age of 58 years (range 22–87). All had newly diagnosed APL (100%) with 62% (n = 52) being of low/intermediate risk according to the Sanz score. Out of these patients 75% (n = 39) received an ATO-ATRA based induction regimen followed by a median of 4 courses of ATO-ATRA consolidation (according to the APL 0406 study). Of 36 patients evaluable for response to induction therapy, 36/36 (100%) patients achieved a complete remission (CR) after ATO-ATRA. Early death rate within 30 days of therapy was 0%. After a median follow-up of 12 months, the event-free survival, cumulative incidence of relapse and overall survival at 12 months for these patients were 100%, 0% and 100%, respectively. Therapy was well tolerated and no new safety signals have been obtained. These real world data from a prospective German registry provide further evidence for the safety and anti-leukemic efficacy of ATO-ATRA in low/ intermediate risk APL. These results further support ATO-ATRA as the new standard of care in this clinical setting. Disclosure: Uwe Platzbecker: Expert Testimony: Research grant by TEVA Pharmaceuticals Richard Schlenk: Expert Testimony: Research grant by TEVA Pharmaceuticals

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Multiples Myelom V359

The new definition of multiple myeloma Ludwig H.P.1 Wilhelminen Cancer Research Institute, Center for Oncology, Haematology, Vienna, Austria 1

Symptomatic multiple myeloma usually evolves from a precursor state, namely from monoclonal gammopathy of undefined significance (MGUS) or from smoldering multiple myeloma (SMM). Several risk factors and algorithms for identifying patients at higher risk for transformation from MGUS and SMM to symptomatic myeloma have been developed. In addition, to these parameters, the following criteria for ultra-high-risk for rapid progression to symptomatic multiple myeloma have been established: SMM with bone marrow plasma cell infiltration of ≥60%, or with a free light chain ratio of ≥100, or with presence of >1 focal bone lesion of ≥5mm detected by MRI scan. Patients with these characteristics have a transformation rate after 2 years of 62–95%, 72–76% and 55–62%, respectively. These new criteria are now, in conjunction with the traditional CRAB criteria, summarized under the acronym SLiM CRAB. The high progression rate of patients with ultra-high-risk SMM prompted the International Multiple Myeloma Working Group to designate these patients ‘early myeloma’ in which active anti-myeloma therapy should be initiated. This recommendation is based on the results of a small prospective randomized trial in patients with high risk SMM. Patients were assigned either to active therapy with lenalidomide-dexamethasone given for nine cycles followed by lenalidomide maintenance therapy or to control (Quiredex trial, Mateos MV et al., NEJM 2013). Results showed a significant increase of PFS and OS in the active therapy cohort. However, due to some limitations of this trial confirmation of these results in a study applying modern imaging technology such as whole body CT, or MRI, or PET/CT and free light chain monitoring of patients (which was not available in the Quiredex trial) seems desirable. Indeed, several trials presently are ongoing or planned in this patient population, but only one compares early treatment with an untreated control group.

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Although these recommendations for initiation of early therapy in patients with ultra-high-risk SMM have been widely distributed, clinical reasoning should be the basis for decision making in those patients. Given the substantial number of ultra-high-risk SMM patients with ≥1 small focal lesion, or a free light chain ratio of ≥100 not having progressed after 2 years (38–45%, and 24–28%, respectively), a watch-and-wait strategy with short follow up intervals may be a reasonable alternative for carefully selected patients. Disclosure: Heinz Ludwig: Financing of Scientific Research: Vortragshonorare und Honorare von Advisory Boards; Expert Testimony: Takeda V360

First-line therapy Einsele H.1 University Hospital Würzburg, Internal Medicine II, Würzburg, Germany

In younger patients 70–75 years in Germany, we generally evaluate the suitability of the patient for high dose chemotherapy and autologous stem cell transplantation. If there are no significant comorbidities the patient – preferentially in a clinical trial – will be treated with high dose chemotherapy and autologous stem cell transplantation. The induction treatment should avoid the use of melphalan and long term exposure to IMiDs to allow a sufficient collection of stem cells. The induction treatment prior to stem cell collection should include at least one of the novel agents, currently the combination of two novel agents is tested to increase response quality prior to stem cell transplantation. The currently most successful induction treatment for patients undergoing autologous stem cell transplantation is the combination of carfilzomib, revlimid and dexamethasone which in studies in the US has been shown to be associated with low toxicity and often induces a VGPR or even complete remission already prior to transplantation. Currently there is a trend to go for tandem transplantation because several studies and also meta-analyses have shown that especially in high risk patients progression free and overall survival is significantly prolonged after tandem transplantation. Several study groups have shown that in patients with a suboptimal response after stem cell transplantation, consolidation can improve response quality but also improve progression and overall survival. Thus, consolidation treatment after autologous stem cell transplantation is increasingly used and already standard in France. Allogeneic stem cell transplantation is only offered as first line treatment in patients with ultra high risk criteria like patients with 17p deletion, extramedullary disease or high serum LDH. For patients who are not suitable for a stem cell transplantation there are currently two treatment regimens that are widely applied. One is the combination of bortezomib with melphalan/prednisolone mostly applied over 9 cycles or the combination of revlimid and dexamethasone according to the first study administered until progression. Disclosure: Hermann Einsele: Advisory Role: Janssen, Celgene, Amgen, Novartis; Financing of Scientific Research: Janssen, Celgene, Amgen, Novartis; Expert Testimony: Janssen, Celgene Amgen V361

Therapy of relapsed and relapsed refractory multiple myeloma Driessen C.1 Kantonsspital St. Gallen, St. Gallen, Switzerland

Current advances in the drug treatment of multiple myeloma (MM) allow delivering active therapy to the majority of MM patients. However, despite a consistently increasing proportion of patients with high quality remissions after first line therapy, most MM patients ultimately relapse and require subsequent treatments for relapsed and relapsed/refractory MM. The recent approval of next generation drugs like carfilzomib, ixazomib, panobinostat, daratumumab and elotuzumab have led to an unprecedent-

Abstracts

ed complexity of therapeutic options in this setting. The increasing clonal diversity, genetic complexity and redundant signaling especially in R/R MM highlight the ubiquitin proteasome system and the immunophenotype as key therapeutic targets, which are conserved in MM requiring advanced line treatment. We shall here briefly review the biological principles of R/R MM and then relate the mechanisms of action, clinical data and biological-pharmacological properties of available drugs to this biology. Based on this, we shall attempt to rationally position currently available options for drug therapy of R/R MM with regard to sequence, combinations and patient as well as disease characteristics. Disclosure: Christoph Driessen: Advisory Role: Celgene, Amgen; Financing of Scientific Research: Celgene, Amgen, Bristol-Meyers Squibb, Novartis, Takeda, Mundipharma; Expert Testimony: Amgen, Novartis, Celgene, Mundipharma

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Pankreaskarzinom V367

Evidence-based treatment options in metastatic pancreatic adenocarcinoma Böck S.1 Klinikum der Universität München – Campus Großhadern, Medizinische Klinik und Poliklinik III und Comprehensive Cancer Center, München, Germany 1

Since its introduction in the late 90ies, the nucleoside analogue gemcitabine has become the main backbone for systemic treatment of pancreatic ductal adenocarcinoma. For patients with metastatic disease, a large number of phase III trials investigating gemcitabine plus a second drug failed to show an improvement with regard to overall survival. The first trial that found a statistically significant, albeit clinically marginal, survival benefit was the PA.3 study that compared single-agent gemcitabine to gemcitabine in combination with the oral anti-EGFR tyrosine kinase inhibitor erlotinib. In that study, the benefit of adding erlotinib was mainly restricted to patients that developed skin rash during treatment with the EGFR-targeting agent. More recently, the global phase III MPACT study showed a significant increase in response rate, progression-free and overall survival for the combination of gemcitabine with nab-paclitaxel in comparison to gemcitabine alone. Based on these data, nab-paclitaxel received regulatory approval for the first-line treatment of metastatic pancreatic cancer in 2014. With the clinical investigation of FOLFIRINOX (5-fluorouracil [5-FU], folinic acid [FA], oxaliplatin and irinotecan) a gemcitabine-free regimen was introduced in the clinic based on the French randomized phase III Prodige 4 - ACCORD 11/0402 study comparing FOLFIRINOX to gemcitabine in a selected patient population (e. g. ECOG performance status 0 to 1, Bilirubin < 1.5 x ULN, no significant cardiovascular co-morbidities) with metastatic pancreatic adenocarcinoma: FOLFIRINOX significantly increased response rate, progression-free and overall survival and – of note – also quality of life. Both regimens (FOLFIRINOX and gemcitabine + nab-paclitaxel) are currently investigated also in the adjuvant, neoadjuvant and locally advanced disease settings. Up to now, there is no evidence for the efficacy of other molecular targeted drugs than erlotinib in the 1st-line treatment of metastatic pancreatic adenocarcinoma; clinical studies using checkpoint inhibitors are currently ongoing. During the last years increasing evidence supporting the use of 2nd-line chemotherapy after failure of 1st-line gemcitabine-based treatment was generated: two randomized phase III studies were able to find an improvement in overall survival with the use of 5-FU/FA either in combination with oxaliplatin (CONKO-003) or nanoliposomal irinotecan (NAPOLI-1) compared to a reference arm with 5-FU/FA only. Disclosure: Stefan Böck: Advisory Role: Baxalta, Celgene; Financing of Scientific Research: Celgene, Roche; Expert Testimony: Celgene, Roche, Clovis Oncology

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Wissenschaftliches Symposium Wie teuer dürfen Arzneimittel sein? V370

Costs of drugs in oncology. The cases of chronic myelogenous leukemia (CML) and immunotherapy Hochhaus A.1 Universitätsklinikum Jena, Abt. Hämatologie/Onkologie, Jena, Germany

The development of BCR-ABL tyrosine kinase inhibitors (TKIs) has revolutionized CML therapy and allowed patients over the past 15 years to live healthy and productive lives. Imatinib is considered to be the most successful targeted anticancer agent yet developed given its substantial efficacy in treating CML and other malignant diseases. In addition, it set the stage for a series of ATP competing drugs in oncology. This dramatic clinical benefit has come at a high cost to individual patients and health care systems. The arrival of generic imatinib has the potential to markedly impact the cost of care for CML. For most European Union (EU) member countries, the imatinib patent for CML will expire in 2016. Loss of patent exclusivity opens the market to competition from multiple manufacturers. Physicians’ willingness to prescribe generic drugs is related to patient benefit, considering potential differences in safety and efficacy of the generics. In some countries, anecdotal concerns have been raised that the bioavailability and potency of generic imatinib is not equivalent to the branded drug; however, this may not be applicable to the EU. The use of generic imatinib should also be discussed in the context of improved efficacy and tolerability of second generation TKIs. Reduction of the rate of early transition into advanced disease, faster elimination of the leukemic burden and earlier access to discontinuation attempts based on faster achievement of deep molecular responses are arguments in favor of the use of branded second generation drugs. A shorter treatment duration with best available drugs may help to reduce treatment costs as well. In addition to targeted therapy, immunotherapy is experiencing a fulminant renaissance with the introduction of antibodies and small molecules activating the patient’s immune system against tumor neoantigens. These new treatments could radically change oncology, but the question remains whether immunotherapies will be affordable for the treatment of all patients with approved indications. What is a fair price for a cancer drug? Certainly a price that would maintain reasonable profits to drug companies, but remain affordable to patients and to the health care systems. Not all cancer drugs are alike, and those with marginal value will not command high prices. The long-term value of individual immunotherapies remains to be seen and with the development of multiple options prices may reach reasonable levels. Disclosure: Andreas Hochhaus: Employment or Leadership Position: UK Jena; Financing of Scientific Research: Novartis, BMS, Pfizer, Incyte (Ariad); Expert Testimony: Novartis, BMS, Pfizer, Incyte (Ariad) V371

Current prices of innovative drugs are too high Ludwig W.-D.1 HELIOS Klinikum Berlin-Buch, Hämatologie, Onkologie, Tumorimmunologie, Palliativmedizin, Berlin, Germany 1

Rapid progress in the fields of tumor biology, genetics, and immunology have greatly improved our understanding of the mechanisms driving tumor formation and growth, and have spurred the development of a large number of innovative cancer drugs and promising drug-based treatment strategies (e.g., immunotherapy). Simultaneously, the launch prices of new anticancer drugs, often with only marginal benefits for their approved indications, have increased over time and their use has contributed to the rising cost of cancer care. Today, about one tenth of the public expenditures for cancer in Europe (about 6–8% of the health care budget) are attributable to the costs of cancer drugs. Cancer drug prices have almost doubled from a decade ago, and nowadays the average price

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of one year treatment with a cancer drug or combination therapies in oncology may exceed € 100,000 per person. A recent survey of the prices of cancer drugs in high-income countries in Europe has disclosed marked variations in ex-factory prices of originator cancer drugs with Germany, Switzerland, and Sweden showing price data in similarly high ranges. In Germany, anticancer drugs represent the drug class with the highest level of spending in 2015 for the statutory health insurances: approx. 5 billions € only for outpatients. This strong upward trend in pricing of cancer drugs represents a significant challenge for our health-care system. Therefore, diverse measures have been proposed to agree upon drug prices according to their real benefits and to optimize reimbursement of cancer drugs. These include cost-effectiveness thresholds, pricing negotiations based on the early benefit assessment of new drugs within the framework of the act on the reform of the market for medicinal products (AMNOG), and programs recently introduced to help physicians, payers, and patients understand the value of new cancer drugs and thus make better choices about their use. The pros and cons of these initiatives will be briefly discussed. In order to ensure affordable drug-based treatment of cancer in the near future additional measures have to be taken which combine the development and availability of promising new cancer drugs with the sustainability of national health systems. Disclosure: No conflict of interest disclosed.

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Noch kurativ oder schon palliativ? Ethik in der Onkologie V374

Indication in advanced cancer. Conceptual analyses and findings from empirical research Schildmann J.1 Ruhr Universität Bochum, Institut für Medizinische Ethik und Geschichte der Medizin, Bochum, Germany 1

There is considerable variation in decisions whether to offer patients with advanced cancer further cancer specific treatment or not. This variation cannot be fully explained by differences concerning the clinical situation or patients’ preferences. In this presentation I will firstly analyse the concept of “indication” and explore value related aspects of indication as one possible explanation for the variation observed in clinical practice. In a second step there will be a summary of findings from empirical research on factors possibly relevant for decisions about treatment in advanced cancer. In the final part I will discuss strategies which may support ethically informed decisions about the offering or limiting of cancer specific treatment in patients with advanced cancer. References: Emanuel EJ, Young-Xu Y, Levinsky NG et al. Chemotherapy use among Medicare beneficiaries at the end of life. Ann Intern Med 2003;138:639–643. Schildmann J, Tan J, Salloch S, Vollmann J: “Well, I think there is great variation...”: a qualitative study of oncologists´ experiences and views regarding medical criteria and other factors relevant to treatment decisions in advanced cancer. Oncologist 2013; 18:90–96. Disclosure: No conflict of interest disclosed. V375

Early integration of palliative care!? Lordick F.1 University Medicine Leipzig, University Cancer Center Leipzig (UCCL), Leipzig, Germany 1

7 years ago ASCO released a statement that was adopted by the Arbeitsgemeinschaft Palliativmedizin of the German Cancer Society: “Palliative cancer care is the integration into cancer care of therapies that address the multiple issues that cause suffering for patients and their families and im-

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pact their life quality. Effective provision of palliative cancer care requires an interdisciplinary team that can provide care in all patient settings, including outpatient clinics, acute and long-term care facilities, and private homes. Changes in current policy, drug availability, and education are necessary for the integration of palliative care throughout the experience of cancer, for the achievement of quality improvement initiatives, and for effective palliative cancer care research. The need for palliative cancer care is greater than ever notwithstanding the strides made over the last decade. Further efforts are needed to realize the integration of palliative care in the model and vision of comprehensive cancer care by 2020.” The publication of the German multidisciplinary and multiprofessional evidence-based and expert-consented (S3) “Guideline for Palliative Care of Patients with Incurable Cancer” is an important step into this direction. Another milestone shall be the planned law for improving hospice and palliative care in Germany (Hospiz- und Palliativgesetz HPG) which was passed in November 2015.In the past, research data in the field of palliative care were scarce. Actions were taken on a more intuitive and empirical basis. Meanwhile, important research results have been published elucidating especially the role of early integration of palliative care into routine oncology. This raises the question who should deliver palliative care to cancer patients. First and foremost, we need to better understand the real burden on our patients, their preferences and care needs during the course of an incurable disease. This is why we started a multidisciplinary research program in the scientific working group (Sektion B) of the German Cancer Society. Eventually, we need to tailor “who shall provide” and “how to deliver” optimized palliative care. The current understanding is that every physician who cares for patients with severe and incurable diseases should have been trained and should be able to deliver general aspects of symptom relieve and palliative care while for complex palliative counceling and care the specialized palliative care physician is needed. Disclosure: Florian Lordick: Advisory Role: Amgen, Biontech, Boston Biomedical, Ganymed, Lilly, MSD, Nordic, Roche, Taiho; Financing of Scientific Research: Vortragshonorare: Amgen, Celgene, Roche, Lilly; Expert Testimony: Böhringer-Ingelheim, GSK, Fresenius Biotech,; Immaterial Conflict of Interests: Reiseunterstützung: Amgen, Bayer, MSD, Roche, Taiho

V377

Communication with patients and caregivers as preparation for palliative care Hübner J.1 Deutsche Krebsgesellschaft, Berlin, Germany

More and more, early integration of palliative care becomes an accepted part of cancer care. After removing essential doubts (increasing of anxiety, destroying hope) by clinical studies and with the growing evidence on the advantages with respect to symptom management, the question is open as to how early integration may be addressed and integrated in daily communication with the patients. This task is demanding as it is more than just transmitting bad news. Hope might be an essential issue in early integration. The fundament of hope depends on the patient´s individual situation of disease as well as his/her perspectives of life. Additional considerations may be valuable. The needs of patients with advanced cancer are autonomy, individuality, self esteem, authenticity, appreciation. The essential message to convey could be: · We respect your needs and values. · The physician who is responsible for you is reliably at your side. · Its is you, whom we put in the center, your preferences and your aims. · This attendance is a comprehensive – Survival, quality of life and meaning are important goals. Disclosure: Jutta Hübner: Employment or Leadership Position: Deutsche Krebsgesellschaft; Advisory Role: Verschiedene Sozialgerichte

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Schwangerschaft und Krebs V381

Fertility after radio-chemotherapy Dirksen U.T.1, IGHG

V376

More than quality of life assessment: How to value the usefulness of palliative care interventions in patients suffering from cancer Alt-Epping B.1 Universitätsmedizin Göttingen, Göttingen, Germany

Palliative Care has emerged from a „philosophy based on attitudes and skills“ (Saunders 2001) to an independent medical field with an according, framing, structural context. Since then, at the latest, similar standards, medical and economical, have to be applied on palliative care institutions (wards, consultation / liaison services, outpatient clinics, or specialized palliative home care) like on institutions and structures in any other medical field. The question, though, what characterizes “good” palliative care, how palliative care may be of use, and for whom, and how its usefulness can be determined appropriately, is much more complex to answer in palliative care compared to other „curative“ or interventional fields in medicine. This congess contribution describes the multifaceted therapeutic aims in palliative care (including the patient´s perceptions as well as the family environment´s expectations), the problems to assess these therapeutic aims and the therapeutic effectiveness of palliative care interventions in an operationalizable manner (and thereby challenging current „quality of life“ concepts), and the resulting ethical implications on increasingly claimed standard value rating and benchmarking processes that have now reached the field of palliative care. Disclosure: No conflict of interest disclosed.

Abstracts

Universitätsklinikum Münster, Münster, Germany

Five-year survival of childhood, adolescent and young adult cancer currently exceeds 70%. Late health outcomes research has demonstrated that a significant proportion of survivors experience chronic health sequelae. A prominent concern among young cancer survivors, which may causes profound emotional distress, is the increased risk of gonadotoxicity following specific cancer treatments. In female survivors, the risk of premature ovarian insufficiency (POI) from primary gland failure (premature menopause), is associated with infertility, but also with other serious sequelae secondary to estrogen deficiency, such as osteoporosis, cardiovascular disorders, impaired wellbeing and compromised sexual health.. In male survivors impaired spermatogenesis (ISG) results in reduced or lost fertility. Testosterone deficiency causes delayed or arrested puberty and a range of adverse somatic and psychological consequences in adults. Evidence-based guidelines are the backbone for a counseling interview at the time of diagnosis and a well-directed surveillance. The International Guideline Harmonization Group organized an endeavor to critically examine evidence and harmonize long-term follow-up guidelines. The working groups comprised experts from nine countries, chosen in view of their expertise and publications in endocrinology, andrology, pediatric and adolescent hematology/oncology, urology, gynecology, radiation oncology, psychology, epidemiology and guideline methodology. Systematic literature research was performed on publications from 1999 to 2014. Published data on the impact of single chemotherapeutic agents on female of male fertility was of limited quality. Definitions: POI was defined as amenorrhea < 40 years and two independent elevated FSH levels. ISG as azo- oligospermia (15 x 106 spermatozoa/ml) and/or low inhibin B- levels. POI-risk is increased after very high dose alkylating agents. The POI-risk following radiotherapy to the ovaries is age dependent. The sterilizing dose is 20.3Gy in infants, 18.4Gy at age 10 years, and 16.5Gy at

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age 20 years. Male infertility is associated with the treatment of high dose alkylating agents, total body irradiation and radiotherapy involving the testes of 2–3Gy. Following from this, an European consortium is making an attempt to gain more evidence- based data on the impact of systemic treatment on fertility under consideration of patient- derived individual hereditary risk factors. Disclosure: No conflict of interest disclosed.

Freier Vortrag Mammakarzinom V384

Determinants of long term quality of life in breast cancer patients: Results from the clinical cancer registry of the OCC Stuttgart Meisner C.1, Heidemann E.2, Rössle S.2, Henke D.1, Brinkmann F.2 Universitätsklinikum Tübingen, Institut für Klinische Epidemiologie und angewandte Biometrie, Tübingen, Germany, 2Onkologischer Schwerpunkt Stuttgart (OSP), Stuttgart, Germany 1

Introduction: Health related quality of life (QoL) of breast cancer patients after curative surgical treatment is an important endpoint in clinical trials but also in health services research. 2711 patients of the four breast cancer centers of the OCC Stuttgart were supported by a special service during a symptom oriented follow-up care program. These patients were asked to participate on periodic surveys for QoL. Aim of this paper was to describe long-term course of QoL and determine predictors of QoL in breast cancer patients. Methods: A total of 2711 breast cancer patients were tested with the QoL instrument EORTC QLQ-C30. The patients were contacted by the special service by mail every 3 months. All domains of EORTC QLQ-C30 were estimated at different time intervals during their follow-up care up to five years and compared with a reference group of healthy German women. Results: The breast cancer patients reported worse mean scores in all domains of EORTC QLQ-C30 at time of diagnosis. The functional scores increase in the first year of follow-up but not reaching the level of the healthy controls. Only marginal increases were seen for the functional scores of most of the patients after one year of follow-up care. The symptom scores decrease in the first year of follow-up. The scores for nausea/ vomiting and appetite loss reach the level of the healthy controls. The greatest deficits are reported for fatigue, pain, dyspnea and insomnia also in long term. The differences between patients and healthy controls were smaller for age groups >60 years. Therapy-related and tumor biological factors had only marginal influence on QoL. Conclusions: Data collection and analysis of quality of life data is an integral part of the routine in the Oncological Comprehensive Center Stuttgart. Data on the course of the domains of QoL according to EORTC QLQ-C30 for short and long term and analyses for determinants of QoL will be presented. They show a huge variability. Key Words: Breast cancer · Quality of Life · Follow-up Disclosure: No conflict of interest disclosed.

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Patient-reported outcomes of 905 patients with breast cancer two years after start of curative systemic treatment Marschner N.1, Trarbach T.2, Dörfel S.3, Meyer D.4, Müller-Hagen S.5, Zaiss M.6, Boller E.7, Kruggel L.7, TMK Registry Group Praxis für interdisziplinäre Onkologie & Hämatologie, Freiburg i.Br., Germany, Klinikum Wilhelmshaven gGmbH, Wilhelmshaven, Germany, 3Onkozentrum Dresden/Freiberg, Dresden, Germany, 4OSP Göttingen Dres. Meyer / Ammon / Metz, Göttingen, Germany, 5Hämatologisch-Onkologischer Schwerpunkt, Hamburg, Germany, 6Praxis für interdisziplinäre Onkologie & Hämatologie, Müllheim, Germany, 7iOMEDICO AG, Freiburg i.Br., Germany 1 2

Introduction: Patients (Pts) with localized, operable breast cancer have to deal with the psychological strain of having cancer as well as with side effects during therapy. However, what is their situation 2 years (yrs) after start of the systemic antineoplastic treatment? MaLife prospectively investigates Patient-reported-outcomes (PROs) to better understand the real-life situation of these pts during and after treatment. Patients and methods: MaLife is conducted with the Tumour Registry Breast Cancer II (TMK II), an open, prospective, multicenter, observational study of pts recruited at the start of (neo)adjuvant or palliative systemic therapy in Germany. Besides documentation of medical data pts regularly receive a set of questionnaires for 5 yrs. Here, data on 905 pts recruited from more than 100 sites and their PROs (FACT-G, EORTCQLQ-BR23, FACT-Taxane, HADS-D and specifically developed questionnaires) are presented. Results: Median age at start of systemic therapy was 56 yrs. Endocrine therapy (ET) was documented for 58% of pts, 93% of pts received chemotherapy. For all pts observed ≥ 2 yrs, questionnaire return rate was 78% 2 yrs after start of therapy. For 5% of pts relapse or death was documented and therefore questionnaire shipping was stopped. 2 yrs after start of therapy the global quality of life slightly improved (FACT-G, +4%), mainly because of an increase in the physical and functional well-being subscores (+12% each). Future perspective increased by 15% (EORTC-QLQ-BR23 subscale). Systemic therapy side effects (EORTC-QLQ-BR23 subscale) and gastrointestinal symptoms, i.e. obstipation and lack of appetite (specific items), decreased by at least 10%. Pts still reported about neuropathic symptoms (FACT-Taxane, mean change from start of therapy to 2 yrs: +9%), pain in neck, shoulders, back and hips (specific items, +9–11%) and problems in memorizing (specific item, +12%). Pts receiving ET were affected by increased bone pain (specific item, +18%). 31% and 30% of pts showed elevated anxiety at start of therapy and after 2 yrs, respectively. Elevated depression was reported by 29% and 23% of pts at start of therapy and after 2 yrs, respectively (HADS-D). Conclusions: Although 2 yrs after start of therapy pts report better physical and functional well-being and fewer side effects, they still report symptoms of polyneuropathy, pain in the upper body and problems in memorizing. Disclosure: No conflict of interest disclosed. V386

Influence of arm crank ergometry on development of lymphedema in breast cancer patients after axillary dissection Schmidt T.1, Berner J.2, Jonat W.2, Weisser B.3, Röcken C.1, Mundhenke C.2 Krebszentrum Nord, Universitätsklinikum Schleswig-Holstein, Kiel, Germany, Klinik für Gynäkologie und Geburtshilfe, Universitätsklinikum SchleswigHolstein, Kiel, Germany, 3Institut für Sportwissenschaft, Christian-AlbrechtsUniversität, Kiel, Germany 1 2

Introduction: Breast cancer-related lymphedema of the arm can be a long-term sequela of medical treatment and is associated with reduced physical functioning and quality of life. In the past physical activity has been proscribed for women with breast-cancer-related lymphedema. The purpose of this study was to assess the safety and efficacy of an arm crank ergometer (ACE) in breast cancer patients with axillary dissection.

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Method: In a randomized, controlled clinical intervention trial twice-weekly supervised ACE-training was compared with usual care (UC) in 49 breast cancer patients after medical treatment over 12 weeks. Endpoints were change in body composition of the upper extremity measured by bioelectrical impedance analysis (BIA), arm volume (arm circumference), muscular strength (NM), quality of live (QL) (EORTC QLQ C30+BR23) and fatigue (MFI 20) before and after 12 weeks. Results: During the intervention no exacerbate was detected. Patients in the ACE improved significantly in lean body mass (LBM) (OP-arm p: 0.017; control-arm p: 0.004), skeletal muscle mass (ACE p: 0.049; UC p: 0.385) and in a significant decrease in body fat (ACE p: 0.009; UC p: 0.393). In the ACE, as well the UC group a significant increase of the armpit circumference was detected during the training period. The magnitude of the gain was higher in the UC. In all other measured regions of the arm a significant decrease of circumference was seen in both groups. As compared with the control group, the ACE-group had a greater improvement in muscular strength of the upper extremity (ACE p: n.s.; UC p: n.s.). In both groups a trend for improvement of QL and general fatigue (ACE p: n.s.; UC p: n.s.) was seen. In the ACE-group a significant improvement in physical functioning (ACE p: 0.39; UC p: 0.355) and a decrease in physical fatigue (ACE p: 0.001; UC p: 0.519) were detected. Conclusion: The results confirm the safe execution of the training with an ACE and highlight improvements in strength, QL and reduced symptoms. All authors declare no conflict of interest. Disclosure: No conflict of interest disclosed. V387

Mcl-1 inhibition as a novel approach for personalized breast cancer therapy Bashari M.H.1,2, Malvestiti S.1, Fan F.1, Vallet S.1, Sattler M.3, Cardone M.H.4, Opferman J.T.5, Jäger D.1, Podar K.1 National Center for Tumor Diseases (NCT), University of Heidelberg, Medical Oncology, Heidelberg, Germany, 2Faculty of Medicine, Universitas Padjadjaran, Department of Pharmacology and Therapy, Bandung, Indonesia, 3Dana-Farber Cancer Institute, Harvard Medical School, Medical Oncology, Boston, United States, 4Eutropics Pharmaceuticals, Inc., Cambridge, United States, 5St. Jude Children’s Research Hospital, Memphis, United States 1

Introduction: Despite significant therapeutic improvements over the last two decades, breast cancer (BC) remains the leading cause of cancer-related mortality in women. Anti-apoptotic Bcl-2 family members including Bcl-2, Bcl-xL and Mcl-1 are frequently upregulated in solid and hematologic malignancies and thereby violate the programmed cell death. Based on our own and other studies, Mcl-1 levels, in particular, are consistently elevated in Her2-positive and triple negative (TN)BC and correlate with drug resistance and poor prognosis. Mcl-1 therefore represents a promising target for BC therapy. Methods: The anti-BC cell-activity of the novel small molecule Mcl-1 inhibitor EU5346 was evaluated alone or in combination with other antiapoptotic agents, hormone therapy, targeted therapy, or chemotherapy using proliferation and spheroid forming assays as well as immunoblotting. Synergistic activity of combination therapies was determined by the Chou-Talalay method. Protein level-based activity of EU5346, the anti-Bcl-2 inhibitor ABT-199 and the anti-BCL-xL inhibitor WEHI-539 was verified in Mcl-1Δ/null versus Mcl-1wt/wt Murine Embryonic Fibroblasts. Results: Using protein profiling and ONCOMINE analysis of our own and publically available data showed that Mcl-1 is consistently overexpressed not only in Her2-positive and TN, but also Luminal A BC cells, while Bcl-2 and Bcl-xL expression is variable. As expected, anti-BC activity of ABT-199 and WEHI-539 in Her2-positive, TN, and Luminal A BC cells was based on Bcl-2 and Bcl-xL protein levels. In order to predict the IC50 of EU5346 in BC cells we additionally developed a mathematical scoring system on the protein levels of Bak, Bim, and NOXA. Synergistic anti-BC activity of low dose EU5346 with ABT-199 or WEHI-539 was observed only in those cells expressing Bcl-2 or Bcl-xL, respectively. Similarly, low dose EU5346 also induced significant anti-BC activity in Lu-

Abstracts

minal A cells, when combined with tamoxifen; in Her2-positive BC cells, when combined with trastuzumab; and in TNBC cells, when combined with paclitaxel. Finally, EU5346 overcame resistance to paclitaxel in paclitaxel-resistant TNBC cells. Conclusions: Our data further support a key role of Mcl-1 in the survival and drug resistance of BC cells. Importantly, they provide the preclinical framework for the personalized, clinical use of Mcl-1-inhibitor-based drug combinations with antiapoptotic, antihormonal or chemotherapies to improve patient outcome in BC. Disclosure: Muhammad Bashari: Expert Testimony: MHB received research support from EUTROPICS Klaus Podar: No conflict of interest disclosed. V388

Novel oncogenic roles of the transcriptional regulator EVI1 in ER- breast carcinoma Wang H.1,2, Schäfer T.1, Konantz M.1, Braun M.3, Perner S.4,5, Varga Z.6, Moch H.6, Reich S.2, Fehm T.7, Kanz L.2, Schulze-Osthoff K.8,9, Lengerke C.1,2,10 University Hospital Basel, Department of Biomedicine, Basel, Switzerland, University of Tuebingen Medical Center II, Department of Hematology & Oncology, Tuebingen, Germany, 3University Hospital of Bonn, Department of Prostate Cancer Research, Institute of Pathology, Bonn, Germany, 4University of Luebeck, Institute of Pathology, Luebeck, Germany, 5The Leibniz Research Center Borstel, Borstel, Germany, 6University Hospital Zurich, Institute of Surgical Pathology, Zurich, Switzerland, 7University Hospital Duesseldorf, Women´s Hospital, Duesseldorf, Germany, 8University of Tuebingen, Interfaculty Institute of Biochemistry, Tuebingen, Germany, 9German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Heidelberg, Germany, 10 University Hospital Basel, Clinic for Hematology, Basel, Switzerland 1 2

Introduction: Ecotropic viral integration site-1 (EVI1) is a transcription factor, particularly known for its role in acute myeloid leukemia aggressiveness. Aberrant EVI1 expression is also detectable in several solid tumors. Here, we explore EVI1 in breast carcinoma and find that high EVI1 expression particularly impacts prognosis and biology of the estrogen receptor negative (ER-) subset. Methods: EVI1 expression was analyzed by qRT-PCR, immunoblot and respectively immunohistochemistry in 8 human BC cell lines and a tissue microarray (TMA) containing 608 patient samples. EVI1 expression was modulated in BC cell lines and primary cells using lentiviral vectors and effects analyzed in cell cycle, BrdU, apoptosis, tumor sphere, migration and in vivo tumorigenicity (zebrafish and mouse) xenotransplantation assays and molecularly by microarray gene expression and ChIP assays. Results: EVI1 expression was detectable in ~80% of BC samples, independent of their ER status; however, high expression associated with shorter overall survival in ER- (p = 0.028) but not ER+ (p = 0.307) patients. Functionally, EVI1 downregulation impaired BC cell proliferation by inducing a G0/G1 arrest and enhancing apoptosis. Furthermore, EVI1 knockdown suprpessed tumor sphere formation and migration of BC cells. Gain-of-function studies using EVI1 overexpression induced opposite phenotypes confirming these data. Moreover, EVI1 knockdown suppressed in vivo tumorigenicity of human BC cells xenotransplanted in fish and mice. Interestingly, treatment with β-estradiol rescued growth and in vivo tumorigenicity of ER+ (but not ER-) EVI1 knockdown BC cells. Molecularly, EVI1 induced the MAPK-pathway and the expression of GPCR signaling associated genes such as KISS1 which could be identified by ChIP as a direct EVI1 target. In line, stimulation with the KISS1 ligand Kisspeptin induced MDAMB231 cell migration, and functionally rescued migration in EVI1 knockdown ER- but not ER+ BC cells. Conclusion: EVI1 expresses in a majority of breast carcinoma samples independent of their ER status. However, in the presence of estrogen, EVI1 selectively impacts the biology of ER- tumors where it drives cell growth via MAPK and migration via KISS1. Although not selectively marking breast cancer stem cells (CSCs), EVI1 expression appears to co-regulate this compartment. Prospective studies are needed to analyze the importance of EVI1 as a biomarker in breast carcinoma. Disclosure: No conflict of interest disclosed.

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V389

The role of FGFR1 signaling in breast cancer to induce tumor angiogenesis

Freier Vortrag

Niedrigmaligne B-Zell-Lymphome – Klinik

Golfmann K.1, Meder L.1, Schuldt P.1, Ullrich R.1,2

V390

University of Cologne, Cancer Therapy and Molecular Imaging, Clinic I for Internal Medicine and Centre for Molecular Medicine Cologne (CMMC), Cologne, Germany, 2University Hospital of Cologne, Center for Integrated Oncology, Cologne, Germany

Two years Rituximab maintenance vs. observation after first line treatment with bendamustine plus rituximab (B-R) in patients with mantle cell lymphoma: first results of a prospective, randomized, multicenter phase 2 study (a subgroup study of the StiL NHL7-2008 MAINTAIN trial; Clinicaltrials.gov Identifier: NCT00877214)

The phase I study of BGJ398, a potent, selective pan-FGFR inhibitor, demonstrated no clinical efficacy in breast cancer harboring amplification of FGFR1. In contrast, Lucitanib and also Dovitinib, dual FGFR and VEGFR inhibitors, demonstrated promising results in patients with FGFR1 amplified breast cancer. Hence, FGFR1 amplification seen in breast cancer may not be as much of a driver event in these patients. We here aim to decipher the role of FGFR1 for tumor angiogenesis and to identify a possible feedback mechanism in breast cancer. Preclinical study with BGJ398, Dovitinib and BAY1 (a selective FGFR inhibitor from BAYERHealthcare) on FGFR1 amplified breast cancer cell lines. Copy number and expression profile were prior tested. Multiplex ELISA, Western Blot analysis and knockdown experiments were performed to unravel the molecular downstream pathway. All in vivo studies were conducted with an orthotopic breast cancer model and tumor growth was measured by BLI_µCT. Breast cancer cell lines MDA-MB-134, CAL-120 and JIMT-1, carrying the amplified FGFR1 gene, were not sensitive to selective FGFR1 inhibition in vitro and knockdown of FGFR1 with shRNA does not repressed cellular growth. Nevertheless, BGJ398 and BAY1 effectively inhibited phosphorylation of FGFR1 and downstream MEK/ERK signalling without deregulating the AKT pathway. We demonstrated that FGFR1 amplified breast cancer cells upregulate VEGF-A expression and secretion under FGF-2 stimulation via activation of PI3K/AKT signalling cascade. Only a combination of FGFR1 and PI3K- inhibition causes significant VEGF-A reduction. We also demonstrated that FGFR1 overexpression was positively correlated with vessel density and tumour growth in vivo. Finally, Dovitinib treatment, in contrast to BGJ398, resulted in significant tumour growth inhibition in FGFR1-amplified orthotopic breast cancer xenografts The results underscore the angiogenic role of FGFR1-mediated signaling in breast cancer. They provide important insights into FGFR1 signaling and a possible feedback mechanism in FGFR1 amplified breast cancer patients. Disclosure: No conflict of interest disclosed.

Rummel M.1, Knauf W.2, Goerner M.3, Söling U.4, Lange E.5, Hertenstein B.6, Eggert J.7, Schliesser G.C.8, Weide R.9, Blumenstengel K.10, Detlefsen N.11, Hinke A.12, Kauff F.13, Barth J.13, Studiengruppe indolente Lymphome (StiL) Justus-Liebig-Universität, Universitätsklinik, Med. Kl IV, Gießen, Germany, Onkologische Praxis, Frankfurt, Germany, 3Städtisches Klinikum, Bielefeld, Germany, 4Onkologische Praxis, Kassel, Germany, 5Evangelisches Krankenhaus, Hamm, Germany, 6Klinikum Bremen Mitte, Hämatologie und Onkologie, Bremen, Germany, 7Onkologische Praxis, Moers, Germany, 8Onkologische Praxis, Gießen, Germany, 9Praxis für Hämatologie und Onkologie, Koblenz, Germany, 10 Praxis für Hämatologie und Onkologie, Eisenach, Germany, 11Justus-LiebigUniversität, Universitätsklinik, Gießen, Germany, 12WiSP, Wissenschaftlicher Service Pharma GmbH, Langenfeld, Germany, 13Justus-Liebig-Universität, Universitätsklinik, Hämatologie, Med Kl. IV, Gießen, Germany 1 2

Introduction: Rituximab maintenance is part of a standard treatment approach for follicular lymphoma. In mantle cell lymphoma (MCL), however, it is not yet common practice. In this study we compared the effect of rituximab maintenance vs observation after first-line treatment with B-R in patients with previously untreated MCL. Methods: Patients were required to have stage II (with bulky disease >7 cm), III, or IV disease for registration in this study. Primary endpoint was progression free survival (PFS). Secondary endpoints included response rates, overall survival (OS), time to progression, event free survival, toxicity. Patients were treated with up to 6 cycles of B-R plus 2 additional cycles rituximab. 122 Patients who have responded to B-R were then randomized to either rituximab maintenance (375 mg/m2 every 2 months for a total of 2 years) or observation only. Results: A total of 118 patients were evaluable for the analysis, 58 (49%) were randomized to maintenance with rituximab and 60 (51%) to observation, respectively. Patient characteristics were comparable for both groups. Median patient age was 71 years, median time of observation was 57,7 months at the time of this analysis (April 2016). No statistical significant differences in PFS between both arms could be observed (p = 0.211, 49 events, HR 0.70, 95% CI 0.40 - 1.22). The median PFS for R maintenance was 74.8 months (95% CI 65.4 - nyr). For the observation arm the median PFS was 54.7 months (95% CI 40.1. - n. y. r.). The results for overall survival showed no difference (p = 0.338, 27 events, HR 1.45, 95% CI 0.68 - 3.10) with a median of nyr for both arms. Conclusions: After a median observation time of 4.8 years from 1st cycle B-R, the results are yet inconclusive. Up to date we were not able to demonstrate a statistical evidence supporting the benefit of R maintenance after B-R in the treatment of patients with MCL. Longer follow-up is needed before final results can be presented. Disclosure: Mathias Rummel: Advisory Role: Mundipharma, Roche; Financing of Scientific Research: Mundipharma, Roche Jürgen Barth: Financing of Scientific Research: Mundipharma, Roche

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Single-agent Ibrutinib is efficacious and well tolerated in Rituximab-refractory patients with Waldenström’s Macroglobulinemia (WM): Initial results from an international, multicenter, open-label phase 3 substudy (iNNOVATE™) Buske C.1, Trotman J.2, Tedeschi A.3, Matous J.V.4, Macdonald D.5, Tam C.6, Tournilhac O.7, Ma S.8, Oriol A.9, Heffner L.T.10, Shustik C.11, Garcia-Sanz R.12, Cornell R.F.13, Fernández de Larrea C.14, Castillo J.J.15, Granell M.16, Kyrtsonis M.-C.17, Leblond V.18, Symeonidis A.19, Singh P.20, Li J.20, Graef T.20, Bilotti E.20, Treon S.15, Dimopoulos M.A.21, on behalf of the iNNOVATE™ investigators Comprehensive Cancer Center Ulm, Institute of Experimental Cancer Research, University Hospital of Ulm, Ulm, Germany, 2Concord Hospital, University of Sydney, Concord, Australia, 3Niguarda Ca’ Granda Hospital, Milan, Italy, 4 Colorado Blood Cancer Institute, Denver, United States, 5Dalhousie University, Halifax, Canada, 6Peter MacCallum Cancer Centre & St. Vincent’s Hospital, Melbourne, Australia, 7Hématologie Clinique Adulte et Thérapie Cellulaire, CHU Clermont-Ferrand, Clermont-Ferrand, France, 8Robert H. Luri Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, United States, 9Institut Català d’Oncologia, Hospital Germans Trias i Pujol, Barcelona, Spain, 10Winship Cancer Institute of Emory University, Atlanta, United States, 11Royal Victoria Hospital at McGill University Health Centre, Montreal, Canada, 12Hospital Universitario de Salamanca, Salamanca, Spain, 13VanderbiltIngram Cancer Center, Nashville, United States, 14Hospital Clínic de Barcelona, IDIBAPS, Barcelona, Spain, 15Dana-Farber Cancer Institute, Boston, United States, 16 Hospital Sant Pau, Barcelona, Spain, 17National and Kapodistrian University of Athens, School of Medicine, Athens, Greece, 18Hôpital Pitié-Salpêtrière APHP, UPMC Université Paris, Paris, France, 19University of Patras Medical School, Patras, Greece, 20Pharmacyclics LLC, an AbbVie Company, Sunnyvale, United States, 21National and Kapodistrian University of Athens School of Medicine, Athens, Greece 1

Introduction: Single-agent ibrutinib (ibr) demonstrated durable responses in previously treated patients (pts) with WM (Treon, NEJM 2015). We report efficacy and safety of ibr in WM pts refractory to their last rituximab-containing therapy (relapse after < 12 months or failure to achieve at least a minor response). Methods: Pts with confirmed diagnosis of WM, rituximab refractory, symptomatic disease requiring treatment (Kyle 2003), platelets ≥50,000 cells/mm3, and absolute neutrophil count ≥750 cells/mm³ received oral ibr 420 mg daily continuously until progressive disease (PD) or unacceptable toxicity. Study endpoints included PFS, ORR, OS, and hemoglobin (Hgb) improvement. Results: Among 31 treated pts, median age was 67 years (range, 47–90); 42% had a high International Prognostic Score System for WM, with a median of 4 prior therapies (68% with ≥3 prior therapies) and ECOG 2 in 19%. In addition to rituximab, the most common prior therapies were corticosteroids and alkylating agents (81% each). Median Hgb and IgM at baseline were 10.3 g/dL and 3830 mg/dL, respectively. Any-grade adverse events (AEs; >15%) included diarrhea (42%); upper respiratory tract infections, hypertension, increased bruising (23% each); nausea, thrombocytopenia, neutropenia (19% each); and pyrexia, arthralgia, back pain (16% each). Common grade ≥3 AEs included neutropenia (13%), hypertension (10%), anemia and diarrhea (6% each). Serious AEs occurred in 29% pts. Four pts discontinued ibr—2 due to PD and 2 due to an AE (gastrointestinal AL amyloidosis and diarrhea); 27 (87%) continue ibr. No IgM flare, atrial fibrillation or major bleeding occurred. Four of 5 pts had no need for plasmapheresis for disease control at the end of cycle 1. ORR at a median follow-up of 12.7 months was 84% (major response rate [MRR; ≥partial response] of 68%), and the estimated 1-year PFS rate was 93%. Baseline median Hgb increased to 12.8 g/dL at Week 49. Baseline median IgM declined by >50% after 1 cycle, with continued improvement over time (median 920 mg/dL at Week 49). Initial analysis of 23 baseline samples identified MYD88MUT/CXCR4WT in 16 pts and MYD88MUT/CXCR4WHIM in 6 pts. ORR and MRR were 88% vs. 83% and 75% vs. 83%, respectively. One of the 23 pts had MYD88WT/CXCR4WT with a best response of stable disease.

Abstracts

Conclusions: Single-agent ibr is highly active (84% ORR) and well tolerated in the rituximab-refractory WM population and presents an attractive treatment option in this patient population. Disclosure: Christian Buske: Employment or Leadership Position: N/A; Advisory Role: Janssen, Roche, Gilead; Stock Ownership: N/A; Honoraria: N/A; Financing of Scientific Research: Roche, Celgene, Janssen; Expert Testimony: Roche, Janssen; Other Financial Relationships: Travel, accommodations, expenses: Roche, Celgene, Janssen; Immaterial Conflict of Interests: N/A Meletios Dimopoulos: Employment or Leadership Position: N/A; Advisory Role: Janssen, Celgene, Onyx, Amgen, Novartis; Stock Ownership: N/A; Honoraria: N/A; Financing of Scientific Research: Janssen, Celgene, Onyx, Amgen, Novartis; Expert Testimony: Genesis Pharma; Other Financial Relationships: N/A; Immaterial Conflict of Interests: N/A V392

Treatment and outcome patterns in patients with relapsed Waldenström Macroglobulinemia from a large observational Pan-European data platform Buske C.1, Sadullah S.2, Kastritis E.3, Tedeschi A.4, Garcia-Sanz R.5, Bolkun L.6, Leleu X.7, Willenbacher W.8, Hajek R.9, Minnema M.C.10, Cheng M.11, Graef T.11, Dimopoulos M.A.3, on behalf of the European Consortium for Waldenström’s Macroglobulinemia (ECWM) University Hospital Ulm, Ulm, Germany, 2James Paget University Hospital, Norfolk, United Kingdom, 3National and Kapodistrian University of Athens, Athens, Greece, 4Niguarda Ca’ Granda Hospital, Milan, Italy, 5Complejo Asistencial Universitario de Salamanca, Salamanca, Spain, 6Medical University Hospital of Bialystok, Bialystok, Poland, 7Hopital de La Milétrie, CHU, Poitiers, France, 8Innsbruck Medical University, Innsbruck, Austria, 9University Hospital of Ostrava, Ostrava, Czech Republic, 10University Medical Center Utrecht, Utrecht, Netherlands, 11Pharmacyclics LLC, an AbbVie Company, Sunnyvale, United States 1

Introduction: Data on treatment choices and their outcome in patients (pts) outside clinical trials are lacking for Waldenström macroglobulinemia (WM). The project goal was to generate real-world data on diagnosis, treatment patterns, and outcomes for WM over a decade in a large pan-European effort. Methods: Retrospective record reviews were completed for pts who fit the following criteria: confirmed WM, symptomatic disease at treatment initiation, frontline (FL) treatment started between Jan 2000 and Dec 2010, at least 1 salvage regimen, and availability of clinical/biologic evaluation at diagnosis/initial therapy. Endpoints included initial/subsequent treatment choices, progression-free survival (PFS), and overall survival (OS). Results: Charts were reviewed for 368/155 pts across second line (2L)/ third line (3L). Median age at treatment initiation was 63 yr (range, 29–89). Reasons for initiating treatment were cytopenias (77%; anemia [75%]), constitutional symptoms (56%), IgM-related symptoms (55%), and organomegaly (26%). Monotherapy use fell from 35% in FL to 21%/22% in 2L/3L. Age ≥70 was associated with greater FL use of monotherapy (40%) vs combination therapy (29%). Combination therapy with rituximab increased from 36% in FL to 63%/56% in 2L/3L. Across all lines, rituximab and cyclophosphamide were the most common agents, excluding steroids, used as monotherapy or in combinations. Median PFS decreased from 31 mo for FL to 24/16 mo for 2L/3L. Median PFS varied by country and choice of agents but was similar for pts < 50 and ≥50 yr. Median PFS improved for pts who received rituximab in 2L vs pts who did not (26 vs 19 mo, P = 0.014). PFS was similar in 3L with regard to rituximab use (15 vs 16 mo, P = 0.69). FL rituximab did not affect median PFS of subsequent lines (2L: 24 vs 23 mo, P = 0.87; 3L: 13 vs 16 mo, P = 0.12). Median OS was not reached but was significantly lower in pts ≥75 yr (70 mo; P < 0.0001) or in pts with high-risk IPSSWM risk score (91 mo; P = 0.0014). No difference in OS was observed in pts with FL rituximab compared to pts not treated with FL rituximab. Other malignancies were reported in 14% after treatment for WM. Conclusions: For European WM pts, monotherapy is widely used, even at first relapse, with notable differences between countries. Rituximab use was associated with improvement in median 2L PFS. This large dataset

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will promote understanding of WM treatment practices and survival outside of a clinical trial setting. Disclosure: Christian Buske: Advisory Role: Janssen, Roche, Gilead; Financing of Scientific Research: Roche, Celgene, Janssen; Expert Testimony: Roche, Janssen Meletios Dimopoulos: Advisory Role: Janssen, Celgene, Onyx, Amgen, Novartis; Financing of Scientific Research: Janssen, Celgene, Onyx, Amgen, Novartis; Expert Testimony: Genesis Pharma V393

IgH-based Next-Generation Sequencing for MRD detection in follicular lymphoma Herzog A.1, Knecht H.1, Herrmann D.1, Unterhalt M.2, Hiddemann W.2, Kneba M.1, Brüggemann M.1, Hoster E.2, Pott C.1 Medizinische Klinik II, Universitätsklinikum Schleswig-Holstein, Campus Kiel, Kiel, Germany, 2Medizinische Klinik und Poliklinik III, Klinikum der Universität München, München, Germany 1

Introduction: MRD monitoring is of great value to determine the risk of relapse in patients with follicular lymphoma (FL). However, MRD assessment is restricted to those 50%-65% of patients harbouring a PCR detectable t(14;18) rearrangement as marker for sensitive RQ-PCR. Next generation sequencing (NGS) of IGH might overcome limitations of current PCR based approaches. Therefore we applied an optimized IGH-based NGS approach to identify tumor-specific IGH (V-(D)-J) clonotypes and tested the reliability for quantitation of MRD in FL patients. Methods: We selected 94 diagnostic (PM) with 166 follow-up (FU) samples from 86 FL patients of the German Low-Grade Lymphoma Study Group (GLSG) according to the presence of a clonal IGH rearrangement detectable by consensus PCR. Paired-end libraries were prepared and sequenced on Illumina MiSeq (2 x 250bp). FU samples were spiked with predefined copy numbers of internal reference DNA to enable assessment of MRD by normalisation of NGS reads after sequencing. Identification of IGH clonotypes and quantitation of MRD was done with the bioinformatic analysis pipeline Arrest/Interrogate (Bystry et al. 2016, under revision). For validation of NGS results t(14;18) RQ-PCR from 15 patients (18 PM and 35 FU) and IGH ASO-PCR from 15 patients (15 PM and 31 FU) was performed. Results: Of 30 FL Patients investigated by RQ-PCR in parallel, NGS identified an IGH clonotype in 26 PM. In the 4 NGS negative samples RQPCR demonstrated low level MRD below 10-3. 24/57 FU samples were MRD+ and 16 MRD- by both methods. 17 FU were MRD+ by either RQPCR (n = 14) or NGS (n = 3). 9 of the NGS negative samples showed very low-level RQ-PCR MRD (10-5 ). MRD results determined by NGS were well correlated to RQ-PCR values (r2 = 0.599, p < 0.000001).In additional 56 Patients without RQ-PCR assays but qualitative detection of clonal IGH rearrangements using GeneScan, NGS identified an IGH clonotype in 52/56 (92.8% ) of the FL patients. 26/93 FU samples were MRD+ and 67 MRD-. 14 were MRD+ by NGS only, while in 2 samples only GeneScan showed MRD+. Conclusions: The application of IGH-NGS showed robust identification of clonotypes in FL patients allowing reliable and sensitive MRD assessment. However, lymphoma infiltration of PM and somatic mutation of IGH are critical points for identification of the tumor-specific clonotypes by NGS. IGH-NGS has a great potential to complement current MRD methods allowing MRD assessment in the majority of patients. Disclosure: No conflict of interest disclosed.

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1st-line treatment of patients with mantle cell lymphoma: Treatment reality and first outcome data from the German prospective TLN Registry Knauf W.1, Bertram M.2, Fietz T.3, Kirste T.4, Grugel R.5, Schnitzler M.5, Abenhardt W.6, TLN Registergruppe Onkologische Gemeinschaftspraxis, Frankfurt a.M., Germany, 2HämatologischOnkologischer Schwerpunkt, Hamburg, Germany, 3Praxis für Innere Medizin, Hämatologie und Onkologie, Singen (Hohentwiel), Germany, 4Praxis für interdisziplinäre Onkologie & Hämatologie, Freiburg i.Br., Germany, 5iOMEDICO, Freiburg i. Br., Germany, 6MVZ Onkologie im Elisenhof, München, Germany 1

Introduction: Amongst non-Hodgkin lymphomas (NHL), the mantle cell lymphoma (MCL) represents a rather more aggressive type with poor prognosis. For the young patients (pts) guidelines recommend a dose-intensified immuno-chemotherapy followed by autologous stem-cell transplant (ASCT) as first-line treatment (Dreyling, 2014, 2016). However, the majority of pts are elderly (>65 years), and therefore the choice of chemotherapy is influenced significantly by the pts’ general state of health. While clinical trials are restricted to highly selected pts, clinical registries offer insight into real-life treatment and effectiveness. Methods: The open, prospective, clinical registry on lymphoid neoplasms (TLN Registry, ClinicalTrial.gov NCT00889798) documents the treatment of pts with lymphoid B-cell neoplasms by German office-based haematologists. Pts are followed for 5 years (yrs). Data regarding pts and tumour characteristics, co-morbidities, all systemic treatments and response rates, date(s) of progression(s) and date of death are recorded. Between 2009 and 2014, a total of 3,795 pts have been recruited by 122 study sites. Results: Of 1,078 pts with low-grade NHL recruited between 04/2009 and 02/2014 at the start of 1st-line therapy, 139 were diagnosed with / treated for MCL. Pts were median 72yrs old (range 31–92yrs, 74% older than 65yrs), 68% male, 82% presented with tumour stage III/IV (Ann Arbor), 6% with ECOG≥2 and 71% with co-morbidities (7% moderate/severe renal disease, 15% diabetes, 48% hypertension, 7% heart failure, 4% heart infarction). 61% were treated with bendamustine+rituximab (BR), 18% received rituximab+cyclophosphamide+doxorubicine+vincristine+prednisone (R-CHOP); other regimens were used in individual cases only (each ≤2%). Objective response rate (ORR) as assessed by the local site was 87% with the clinical (unconfirmed) complete remission rate of 46%. With a median follow-up of 39 months (mth), prospective median progression free survival (PFSREG) is 62mth (95%-CI 44mth-NA) and 3yrs PFS rate is 65% (95%-CI 54%-73%;). 3yrs overall survival rate (OS) is 76%, while median OS is not reached. 25% of pts have received a 2nd-line therapy. Overall, 13% of pts have been lost to follow-up. Conclusion: In office-based practices in Germany, pts with previously untreated MCL are predominantly treated with BR. First outcome data show that the effectiveness (ORR, PFS and OS) for unselected pts in routine practice is similar to pts treated in trials. Disclosure: Wolfgang Knauf: Advisory Role: Advisory Board Roche, Mundipharma; Financing of Scientific Research: Vortragshonorare Roche, Mundipharma; Other Financial Relationships: Reisekosten Roche, Mundipharma Wolfgang Abenhardt: Employment or Leadership Position: Ärztlicher Geschäftsführer der MVZ; Advisory Role: Advisory Board bei AMGEN, BMS, Celgene, Gilead, Janssen, MSK, Novartis, ROCHE; Other Financial Relationships: Unterstützung des Münchner Lymphom Workshops durch AMGEN, Celgene, Gilead, ROCHE

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CLL2-BIG – a sequential treatment regimen of Bendamustine followed by GA101 and Ibrutinib followed by Ibrutinib and GA101 maintenance in patients with Chronic Lymphocytic Leukemia (CLL): Interim results of a phase II-trial by the German CLL Study Group von Tresckow J.1, Cramer P.1, Bahlo J.1, Engelke A.1, Langerbeins P.1, Fink A.-M.1, Klaproth H.2, Tausch E.3, Fischer K.1, Wendtner C.-M.1,4, Kreuzer K.-A.1, Stilgenbauer S.3, Böttcher S.5, Eichhorst B.1, Hallek M.1 Uniklinik Köln, Klinik I für Innere Medizin/Zentrum für Integrierte Onkologie/ Deutsche CLL Studiengruppe, Köln, Germany, 2Hämatologische/Onkologische Praxis Dr. Schmidt/Klaproth, Neunkirchen, Germany, 3Uniklinik Ulm, Klinik III für Innere Medizin, Ulm, Germany, 4Klinikum Schwabing, Klinik für Hämatologie, Onkologie, Immunologie, Palliativmedizin, Infektiologie und Tropenmedizin, München, Germany, 5Universitätsklinikum Schleswig-Holstein, Campus Kiel, Klinik für Innere Medizin II, Kiel, Germany 1

Introduction: Treatment with anti-CD20 antibodies and targeted drugs show promising efficacy and tolerability in CLL. With the CLL2-BIG trial, a novel combination regimen according to the “sequential triple-T” concept of a tailored and targeted treatment aiming at total eradication of minimal residual disease (MRD) was designed. Methods: Physically fit and unfit CLL-patients were recruited according to a predefined allocation for the two strata of first-line and relapse/refractory treatment. Patients with high tumor burden received two cycles of bendamustine debulking before administration of six cycles of induction treatment with obinutuzumab and ibrutinib. Maintenance therapy with ibrutinib and obinutuzumab every three months followed until achievement of MRD-negative complete remission or up to 24 months. The primary endpoint is overall response rate (ORR) at the end of induction; secondary endpoints include ORR after debulking and safety parameters. Results: 66 patients were included, thereof 31 first-line and 35 relapsed/ refractory with a median number of one prior therapy (range 1–5). The median age was 66.5 (36–85) years and the median CIRS score was 2.5 (0–11). 10 patients had a del(17p). 46 patients received debulking treatment including four patients who stopped study therapy after the first course of bendamustine. 17 patients responded to debulking; in eight patients a (clinical) complete and in nine patients a partial remission was assessed. 21 patients showed a stable disease, six patients progressed. In 20 patients debulking was omitted due to contraindications or low tumor burden. Obinutuzumab was administered to 62 patients of whom 42 received debulking. Occurrence of infusion related reactions (IRR) related to obinutuzumab is shown in table 1. Tab. 1. Infusion related reactions (IRR)

In general, the combination of obinutuzumab and ibrutinib was well tolerated and SAEs were in line with the known toxicity profiles of both drugs. Conclusion: A chemotherapy reduced and targeted regimen is investigated. Our data suggest that bendamustine debulking prior to obinutuzumab might reduce the occurrence of IRRs as compared to historical data [Goede V., 2014]. Disclosure: Julia von Tresckow: Advisory Role: Janssen-Cilag; Expert Testimony: Janssen-Cilag, F. Hoffmann-LaRoche Michael Hallek: Advisory Role: AbbVie, Amgen, Celgene, F. Hoffmann-LaRoche, Gilead, Janssen-Cilag, Mundipharma; Expert Testimony: AbbVie, Amgen, Celgene, F. Hoffmann-LaRoche, Gilead, Janssen-Cilag, Mundipharma

Fortbildung

Migrationsanämien V396

“Migration anemias“- an overview Cario H.1 Klinik für Kinder- und Jugendmedizin, Universitätsklinikum, Ulm, Germany

The recent dramatically increased migration of people from the Middle East and Africa raises particular challenges for all parts of the society including the healthcare system. Diseases which previously did not seem to play role in Germany are brought into the focus of healthcare providers in all medical fields including hematology, facing apparently new, widely unknown or neglected red blood cell disorders. However, due to previous migration processes many disorders here summed up as “migration anemias” have actually been present as serious medical problem in Germany already for longtime. This particularly concerns the hemoglobinopathies, i.e. thalassemias and sickle cell disease, representing the most common monogenic inherited disorders worldwide, together with another “migration anemia”, G6PD deficiency. As compared to other Central European countries, Germany relatively late experienced migration from countries where these disorders are endemic. Rather few patients have been distributed over the whole country. Until the late 1990s, almost exclusively children and adolescents were affected. The heterogeneous origin of patients and various socioeconomic factors did hamper the formation of effective patient organizations. All these factors contributed to a rather low awareness of these disorders leading to their only marginal role in the medical education but also in daily medical practice. Major new aspects in the current situation concern the sudden increase of the number of patients with “migration anemias” needing regular care, many of them affected by severe prior complications. Transfusion therapy is frequently complicated by previous alloimmunization. Excessive iron overload, but also “typical” but “forgotten” symptoms of disorders like thalassemia are now challenging the medical staff. The central allocation of refugees in Germany leads to their presentation to healthcare providers who did not have any contact to patients with “migration anemias” in the past but now have to care for them. At present, the previously claimed centralization of patients with these disorders is not feasible. It has to be replaced by professional networks of pediatric and adult hematologists providing access to adequate medical care for all patients with “migration anemias”, based on existing jointly developed guidelines of DGHO and GPOH and including necessary diagnostic procedures and consultation of specialists experienced in the care of these disorders. Disclosure: No conflict of interest disclosed. V397

Non-transfusion-dependent thalassemia (NTDT) - therapy Dickerhoff R.1 Uni-Klinik Düsseldorf, Pädiatrie Hämatologie/Onkologie, Düsseldorf, Germany

Non-transfusion-dependent-thalassemia used to be called Thalassemia Intermedia, because its severity was generally thought to be in between

Abstracts

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the asymptomatic carriership (Thal. Minor) and the transfusion dependent Thalassemia Major. As NTDTpatients got older it became apparent that this entity can present with a larger spectrum of problems than previously thought and very often morbidity in NTDT is more pronounced than in Thalassemia Major. Also, a wide variety of mutations (both α and ß chain mutations) have been found to cause NTDT. Many NTDT patients go undiagnosed for a long time as they present by hemoglobin analysis as heterozygous ß thalassemias – and nobody thinks of the existence of so called dominant ß thalassemia or looks for a triplication of alpha globin genes accompanying a ß globin mutation . NTDT children with growth failure, widening of marrow space( prominent maxillae)or permanent hemoglobin levels below 6 g/dl need regular transfusions. As patients get older the effects of inefficient erythropoiesis become noticeable: increasing splenomegaly, iron overload (even without transfusions) and extramedullary hematopoiesis (EMH), most often parallel to the spinal column. Splenomegaly , unless extreme or accompanied by severe pancytopenia, should be treated with chronic transfusions as splenectomy carries a high risk for thrombosis. In EMH with neurological symptoms low dose radiation can rapidly decrease the EMH volume while in EMH without neurological impairment transfusions or hydroxycarbamide are indicated to decrease inefficient erythropoiesis. After puberty iron overload has to be looked for and treated with chelation. Transfusions are also necessary in patients with pulmonary hypertension, in some patients with severe HbH disease (non-deletional HbH disease), sometimes during pregnancy, infection or in patients who develop thrombosis despite anticoagulation. Symptomatic cholelithiasis requires cholecystectomy. Leg ulcers are very painful and heal slowly, even with dedicated wound care. Osteopenia and osteoporosis cause very distressing bone pain in many NTDT patients. In some NTDT patients hydroxyurea is useful to decrease transfusion needs. Each NTDT patient is unique and his or her clinical problems have to be evaluated individually. The challenge begins with a correct diagnosis and goes on with a very individualized treatment.

tions. Several surveys among SCD patients and parents showed that 62% accepted a transplant related mortality of >10% and 30% a TRM >30%. GF >10% was acceptable to 64% and >30% for 41%. Chronic GVHD was unacceptable to the majority (80%). Cord blood and haplo-SCT using post-cyclophosphamide (POST-CY) failed in prospective trials due to high graft failure (GF) rates. T-depleted haploidentical HSCT is a well established transplant modality which not only offers a donor for everybody but is also associated with a very low incidence of GvhD. In summary SCD is one of the most prevalent and devastating haematological diseases, barely known in Germany, for which standard of care is insufficient and novel therapeutic options need to be explored in well controlled prospective trials.

Disclosure: No conflict of interest disclosed.

Introduction: Erythroferrone (ERFE) is a regulator of iron homeostasis in the context of hematopoietic stress and erythropoietin (EPO) stimulation as it induces increased iron availability by downregulation of hepcidin. Growth differentiation factor 15 (GDF15) also plays an active but yet to be established role in signaling during ineffective erythropoiesis. In order to investigate potentially disturbed molecular roles of ERFE and GDF15 in Myelodysplastic Syndromes (MDS), we analyzed their gene expression levels in highly purified CD71 positive bone marrow cells derived from MDS patients and healthy controls and correlated the differential expression data with clinical parameters and survival. Methods: CD71+ erythroprogenitor cells of a total of n = 148 MDS, n = 18 sAML patients and n = 35 healthy individuals were isolated immunomagnetically via MACS columns (Miltenyi). After total RNA extraction using the AllPrep DNA/RNA Mini kit (Qiagen), cDNA was transcribed from RNA via Quantitect cDNA synthesis kit (Qiagen). Subsequently, ERFE and GDF15 expression was quantified from cDNA by quantitative PCR. Results: As compared to unselected BM or other specific differentiation lineages, ERFE was almost exclusively expressed in the erythropoietic CD71+ compartment. ERFE expression in the CD71+ subset revealed a highly significant overexpression in MDS IPSS-low/int-1-risk (fold change (FC) = 4.3, p < 0.0001), IPSS-int-2/high-risk (FC = 6.23, p < 0.0001) and sAML (FC = 6.69, p < 0.0001) relative to healthy controls. As compared to healthy controls, GDF15 showed a mean FC = 66 (p < 0,0001). ERFE expression showed no correlation with clinical laboratory parameters. GDF15 levels were positively correlated with plasma iron levels, transferrin saturation and endogenous EPO levels. Both high ERFE and GDF15 expression levels in CD71+ progenitor cells were significantly associated with superior overall survival (ERFE: p = 0.0007, GDF15: p = 0.0001). Conclusion: Highly aberrant overexpression of ERFE and GDF15 in CD71+ erythropoietic progenitor cells suggests an important role for these genes in the dysfunctional erythropoiesis of MDS. The relationship of high ERFE and GDF15 expression with superior survival, especially in low risk MDS patients with no apparent coherence to other established clinical markers warrants further pursuit of ERFE and GDF15 expression

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Sickle cell disease Corbacioglu S.1 Universitätsklinikum Regensburg, Päd. Hämatologie, Onkoloige und Stammzelltransplantation, Regensburg, Germany 1

Hemoglobinopathies are the commonest, life-threatening, monogenic disorders in the world. Over 300.000 newborns with sickle cell disease (SCD) are born annually in Africa and Asia, 100.000 affected patients in the United States. In Europe the prevalence is 15 from 100.000 and there are approximately 3000 recorded SCD patients in Germany, with many additionally unregistered and refugees from high prevalence countries. SCD is overtaking haemophilia and cystic fibrosis in certain parts of Europe. The average life expectancy is between 40 to 50 years. In developing countries, the mortality is between 50%-80% during infancy. SCD is a progressively debilitating and chronic multi-organ disease with a 30% to 50% incidence of disability and unemployment. At 45 years 24% of SCD patients will have developed a stroke, 30% develop silent strokes leading to cognitive impairment in childhood and an impaired quality of life. SCD is the leading cause of stroke in children and adolescents. SCD is a major public health concern. From 1989 through 1993, an average of 75,000 hospitalizations due to SCD occurred in the United States, costing approximately $475 million. Next to future gene therapy which remains disappointing to date, hematopoietic stem cell transplantation (HSCT) is currently the only curative option for SCD. But only 20% have a matched donor available (MUD). This is reflected in the different transplant registries where despite its prevalence only 1200 HSCTs are reported with 75% from matched sibling donors (MSD). Therefore 80% receive currently symptomatic treatment. But medical treatment options besides HSCT cannot cure the disease but only delay the onset of SCD related complica-

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Disclosure: No conflict of interest disclosed.

Freier Vortrag

Myelodysplastisches Syndrom – experimentell V401

Erythroferrone (ERFE) and Growth Differentiation Factor 15 (GDF15) are highly differentially overexpressed in CD71 positive Erythroprogenitor cells of patients with Myelodysplastic Syndromes and associated with survival Mossner M.1, Stöhr A.1, Jann J.C.1, Nolte F.2, Nowak V.1, Obländer J.1, Pressler J.1, Xanthopoulos C.1, Palme I.1, Baldus C.D.3, Schulze T.J.4, Boch T.1, Metzgeroth G.1, Neumann M.3, Hofmann W.K.1, Nowak D.1 Medizinische Fakultät Mannheim der Universität Heidelberg. III. Medizinische Klinik, Hämatologie und Onkologie, Mannheim, Germany, 2St. Hedwig Krankenhaus, Berlin, Germany, 3Charité, Campus Benjamin Franklin, Berlin, Germany, 4Institut für Transfusionsmedizin und Immunologie, Medizinische Fakultät Mannheim der Universität Heidelberg, Mannheim, Germany 1

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profiles in CD71+ BM cells of MDS patients as a possible independent prognostic marker and therapeutic target. Disclosure: No conflict of interest disclosed. V402

Bone homeostasis is altered in a mouse model of myelodysplastic syndrome Weidner H.1, Rauner M.2, Bulycheva E.1, Khandanpour C.3, Bornhäuser M.1,4, Hofbauer L.C.2,4, Platzbecker U.1,4 Department of Medicine I, Technical University, Dresden, Germany, Department of Medicine III, Technical University, Division of Endocrinology, Diabetes, and Bone Diseases, Dresden, Germany, 3Department of Hematology, West German Cancer Center, University Hospital Essen, University DuisburgEssen, Essen, Germany, 4German Cancer Consortium (DKTK), partner site Dresden and German Cancer Research Center (DKFZ), Heidelberg, Germany 1 2

Introduction: Myelodysplastic syndromes (MDS) are a heterogeneous group of clonal diseases that mainly occur in the elderly and are characterized by an ineffective hematopoiesis leading to cytopenias. Increasing evidence suggests the bone microenvironment as a critical modulator to influencing the progression of MDS. However, the underlying mechanisms are not fully understood. NUP98-HOXD13 (NHD13) mice develop MDS-like symptoms at an age of 6 months, including anemia and an increased number of blasts in the bone marrow. Thus, it may be used as model to study bone homeostasis in the setting of MDS. Here, we assessed bone homeostasis in NHD13 mice before (2 month-old) and after (6 month-old) the onset of MDS. Methods: To investigate the bone phenotype we used micro-computed tomography of the femurs of 2- and 6-month-old male NHD13 and wild-type (WT) mice. Bone samples were histologically stained for the osteoclast marker tartrate-resistant acid phosphatase. In addition, serum was analyzed by ELISAs to assess the bone turnover markers C-terminal telopeptide of type I collagen (CTX-I) and procollagen type I propeptide (P1NP). Results: Two-month-old NHD13 mice had already developed leukopenia in the absence of other cytopenias and showed alterations of bone homeostasis. The trabecular bone volume [-26%; P < .05] and trabecular number [-20%; P < .01] were reduced in the femurs of NHD13 compared to WT mice, but the bone resorption marker CTX-I was not changed. Interestingly, the bone formation marker P1NP was increased [3-fold, P < .001]. At the age of 6 months the NHD13 mice had developed anemia indicated by a reduced hematocrit [-17%; P < .001]. The bone volume was unchanged but the trabecular architecture was altered compared to WT mice. Similar to the 2-month-old NHD13 mice, the number of trabeculae was reduced in the 6-month-old NHD13 mice [-15%; P < .001] whereas their thickness was increased [+18%; P < .01]. The histology of NHD13 mice indicated an increased number of osteoblasts [1.5-fold; P < .01] and also the bone formation marker P1NP was elevated [+66%; P < .05] compared to WT mice. Although NHD13 mice had fewer osteoclasts [-77%; P < .01], the bone resorption marker CTX-I was increased by 32% [P < .05]. Conclusion: NHD13 mice show an altered bone homeostasis before and after the onset of MDS. In summary, our study indicates that already minimal alterations in the hematopoietic system have an impact on the bone microenvironment, thus influencing bone homeostasis.

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Iron-overload links to genetic instability in myelodysplastic syndromes Westhofen G.1, Ganster C.1, Beyer F.2, Rassaf T.3, Al-Ali H.K.4, Stuhlmann R.5, Glass B.5, Bacher U.1, Brümmendorf T.2, Germing U.6, Gattermann N.6, Haase D.1 University Medicine Göttingen, Clinics of Hematology and Medical Oncology, Specialized Diagnostic Laboratories INDIGHO, Göttingen, Germany, 2RWTH Aachen University Hospital, Department of Haematology, Oncology, Haemostaseology, and Stem Cell Transplantation, Aachen, Germany, 3HeinrichHeine University Düsseldorf, Department of Cardiology, Pneumology, Angiology, Düsseldorf, Germany, 4University Hospital of Leipzig, Leipzig, Germany, 5Asklepios Klinik St. Georg, Hamburg, Germany, 6Heinrich-Heine University Düsseldorf, Department of Hematology, Oncology, and clinical Immunology, Düsseldorf, Germany 1

Introduction: As numerous experimental as well as clinical studies have shown, iron overload (IO) results in not only a reduced overall survival, but also decreases the leukemia free survival in patients suffering from myelodysplastic syndromes (MDS), yet the related pathomechansims remain unclarified. The aim of our study was therefore to examine and identify associations between genetic instability, oxidative stress and elevated iron parameters in patients suffering from MDS. Methods: 55 patients (pts) with proven MDS have been included in our study, dividing the cohort into two groups according to their serum ferritin (SF) levels, using 275µg/l as a cut-off value for normal vs. elevated SF. All patients were investigated for oxidative stress markers (plasma nitric oxide metabolites; DNA double-stranded breaks (DSB) in CD34+ cells, examined by γH2AX immunofluorescent staining), replicative stress markers (alteration of telomere length (TL) in granulocytes and lymphocytes), and markers for genetic instability (cytogenetic alterations investigated by chromosome banding, SNP-array- and FISH-analyses; molecular mutations, identified by Sanger-sequencing and NGS). Results: Pts. with elevated SF displayed more DSB per CD34+ cell than pts with normal SF levels (median 5.65 [range 2.1–10.8] vs. 1.9 [0.5–6.8], p = 0.016) .Cytogenetic analyses showed an increase in size of the total genomic aberration with elevated SF (median 0 Mbp [0–155] vs. 33.66 Mbp [0–247.9], p > 0.05). TL was significantly reduced in granulocytes of pts with high SF compared to patients with normal SF (median -1.611kb [-4.062–1.311] vs. 0.480kb [-3.127- 5.318], p = 0.006); TL in lymphocytes was not influenced by SF. Not only elevated SF, but also the number of erythrocyte concentrates transfused in the past 3 months showed these correlations with genetic instability markers. Conclusions: Our study, as best to our knowledge is the first to explicitly investigate the link between IO and genome stability, revealed an association of distinct markers for genetic instability with elevated SF and transfusion dependency. Furthermore we were able to show a negative effect of iron at much lower SF levels than previously assumed, starting at the upper normal level of 275µg/l in our clinical chemistry lab. These results further support the thesis of IO being causally related to disease progression and transformation in MDS and should be taken into account for future diagnostics and therapeutic decision-making. Disclosure: Gina Westhofen: No conflict of interest disclosed. Detlef Haase: Financing of Scientific Research: Novartis; Expert Testimony: Novartis

Disclosure: No conflict of interest disclosed.

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–128

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V404

Cytomorphological and molecular background of isolated Y-loss in myelodysplastic syndromes Ganster C.1, Bacher U.1, Germing U.2, Shumilov E.1, Strupp C.2, Shirneshan K.1, Dierks S.1, Stuhlmann R.3, Glass B.3, Bäsecke J.4, SimonBecker S.5, Sievers B.6, Martin R.1, Flach J.1, Harder L.7, Haase D.1 Universitätsmedizin Göttingen, Klinik für Hämatologie und Medizinische Onkologie, Göttingen, Germany, 2Universitätsklinikum Düsseldorf, Klinik für Hämatologie, Onkologie und Klinische Immunologie, Düsseldorf, Germany, 3Asklepios Klinik St. Georg, Klinik für Hämatologie, Onkologie und Stammzelltransplantation, Hamburg, Germany, 4St. Josefs-Hospital, Onkologie und Hämatologie, Cloppenburg, Germany, 5MVZ Alsfeld, Alsfeld, Germany, 6 Onkologische Schwerpunktpraxis, Hildesheim, Germany, 7Institut für Tumorgenetik Nord, Kiel, Germany 1

Introduction: Loss of the Y chromosome (LOY) is a frequent aberration in myelodysplastic syndromes (MDS), observed as a single aberration in 3–4% of male MDS patients (pts.). In previous studies we could show that LOY is not only a phenomenon of higher age in MDS, but may also be a marker for a malignant clone. MDS pts. with isolated LOY have a good prognosis with a very low risk for leukemic transformation. To facilitate an unambiguous discrimination between age-related and clonal LOY, the aim of this study was to identify molecular mutations or cytomorphological features typical for MDS pts. with isolated LOY. Methods: We compared the mutational profile (17 myeloid genes) between 17 pts. with isolated LOY and two pts. with LOY and trisomy 15 and a control group of 197 pts. with MDS and sAML and other cytogenetic abnormalities or normal karyotype. In addition, bone marrow aspirates of 41 pts. with isolated LOY were cytomorphologically evaluated according to the standards of the Duesseldorf MDS registry. Results: The most frequently mutated gene in molecularly analyzed pts. (LOY in ≥50% of metaphases in proven MDS or in ≥75% in suspected MDS) was TET2 (n = 7/19). Further mutations affected CBL (n = 2/15), ZRSR2 (2/15), ASXL1 (1/19) and SRSF2 (1/19). Overall, the proportion of pts. with molecular mutations was lower in the subgroup with isolated LOY compared to the control group (53% vs. 72%). MDS according to WHO criteria (2008) was cytomorphologically confirmed in 36/41 pts. (19x LOY in ≥75% of metaphases, 22x in < 75%). Besides a bone marrow blast count of < 5% in 40/41 cases, a heterogeneous distribution of MDS subtypes was observed. Peripheral blood counts [Hb: mean 10.4 vs. 9.7 g/ dL; WBC: 4.9 vs. 6.0×10(9)/L, thrombocytes: 163 vs. 198×10(9)/L] and dysplasia of the individual cell lines (erythro-, granulo-, megakaryopoesis) did not differ significantly between the groups with a LOY clone size ≥75% and < 75%. Conclusion: MDS with clonal LOY seems to comprise a heterogeneous mixture of cytomorphologic subtypes and to be outlined by marked heterogeneity of the molecular mutation patterns. However, MDS pts. with LOY mostly show absence of marrow blast increase ≥5% and of molecular high risk markers (mutations in TP53, RUNX1, EZH2, ASXL1). Molecular MDS-associated mutations are less frequent in this cytogenetic subgroup as compared to overall MDS. These results are in line with the known favorable prognosis of these pts. according to the IPSS and IPSS-R. Disclosure: No conflict of interest disclosed. V405

Development of RAS-induced zebrafish leukemia models Alghisi E.1, Konantz M.1, Mione M.2, Lengerke C.1,3 Universitätsspital Basel, Department für Biomedizin, Basel, Switzerland, 2Centro di Biologia Integrata, University of Trento, Trento, Italy, 3Universitätsspital Basel, Abteilung für Hämatologie, Basel, Switzerland 1

Background: The zebrafish has emerged as a versatile novel experimental model for studies on developmental hematopoiesis and leukemogenesis. Several oncogenes involved in human leukemia have been successfully overexpressed in zebrafish.

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Aims: We aim to use the zebrafish model to investigate cooperating molecular events involved in leukemia initiation, progression and chemo-resistance. Methods: We took advantage of the Gal4/UAS binary system to overexpress human oncogenic HRAS in zebrafish hematopoietic cells using specific promoters (fli.1, pu.1, runx.1, mpeg1). Blood cell development was studied at different stages by real-time PCR analysis of hematopoietic gene expression, flow cytometry, immunohistochemistry and blood smear morphological assessment. Small molecule library screens are performed on live HRAS overexpressing transgenic larvae and effects assessed by microscopy. Results: Different phenotypes were observed depending on the promoter driving the oncogene expression. HRAS induction via the early hematopoietic promoter fli.1 affected primitive hematopoiesis inducing a myelo-erythroid proliferation and delayed erythrocyte maturation (Alghisi et al. 2013). Alternatively, HRAS expression driven by runx1, pu.1 and mpeg1 did not affect primitive hematopoiesis and allowed studies at later developmental stages. After 1 month, runx1-HRAS fish displayed a cellular expansion of hematopoietic stem/progenitor cells (HSPC) in the kidney marrow (KM), the zebrafish definitive hematopoietic compartment. KM cytospin preparation and flow cytometric analysis confirmed high numbers of undifferentiated cells, indicating that HRAS-overexpression induced HSPC proliferation and impaired differentiation capacity. Similarly, mpeg1-HRAS fish also show increased numbers of blood progenitors in the KM and abnormal gene expression of progenitor markers. Using fli.1-HRAS fish and an FDA approved library comprising around 2000 compounds we have set up an in vivo screen to identify drugs that can suppress the HRAS-mediated hematologic phenotype. Conclusion and outlook: We have established a zebrafish model of HRAS induced hematologic disease. We are currently further investigating this model using in vitro colony forming assays, serial re-transplantations and exploring effects of HRAS in cooperation with other oncogenes. Active compounds identified in the small molecule screen shall be presented at the meeting and further tested for their efficacy in adult disease models. Disclosure: No conflict of interest disclosed. V406

Frequency and clonogenic capacity of the rare CFU-F subpopulation of Mesenchymal Stem Cells (MSC) is significantly reduced in Myelodysplastic Syndromes (MDS) and Acute Myeloid Leukemia (AML) Weickert M.-T.1, Garz A.-K.1, Zwick A.1, Ziegenhain C.2, Grath S.3, Strunk D.4, Enard W.2, Peschel C.1, Oostendorp R.1, Götze K.S.1 Technische Universität München, III. Med. Klinik, München, Germany, 2Faculty of Biology, Ludwig-Maximilians University Munich, Anthropology and Human Genomics Department, München, Germany, 3Faculty of Biology, LudwigMaximilians University Munich, Evolutionary and Functional Genomics, München, Germany, 4Spinal Cord Injury and Tissue Regeneration Center Salzburg (SCI-TReCS), Institut für Experimentelle und Klinische Zelltherapie, Salzburg, Austria 1

Crosstalk of hematopoietic stem cells (HSC) with stromal niche cells is crucial for regulation of HSC function. The stromal niche is derived from mesenchymal stem cells (MSC), giving rise to adipocytes, osteoblasts and chondroblasts. Dsyregulation of niche integrity contributes to leukemogenesis and alterations in MSC function play a prominent role in myelodysplastic syndromes (MDS). We developed a flow cytometric sorting protocol to isolate the rare population of MSC by surface marker expression to study unmanipulated MSC. We asked whether certain MSC subpopulations are dysregulated in MDS and acute myeloid leukemia (AML) and how this may influence HSC behavior. MSC were isolated by flow cytometric sorting from whole bone marrow from patients with newly diagnosed MDS, AML or healthy individuals. Cells were sorted by negativity for CD31/CD235a/CD45 and positivity for CD271. Two distinct populations were identified within the CD31-/CD235a-/CD45-/CD271+ gate: CD271+/CD73+/CD105+/CD90+ (termed +/+) and CD271+/

Abstracts

CONTENTS AUTHOR INDEX

CD73-/CD105-/CD90- (termed ±) cells. To determine CFU-F frequency, cells were seeded in 24-well-plates and formation of CFU-F was observed every second day. CFU-F formation was only observed in the subpopulation of +/+ cells but not in the +/- population. MDS and AML samples showed a slightly reduced +/+ population frequency compared to healthy samples. However, CFU-F capacity in the +/+ population was dramatically impaired in MDS. Proliferation of CFU-F was strongly reduced in MDS samples and only CFU-F forming cells from healthy controls could be expanded subsequently. Differentiation into adipogenic and osteogenic lineages was altered in MDS and AML compared to healthy MSC. Co-culture of healthy CD34+ cells on MSC from MDS and AML showed reduced CFU-C capacity. Gene expression analyses determined that the adipogenic inhibitor DLK1 was significantly down regulated in MSC from MDS. Further RNA sequencing analyses of MSC subpopulations are currently ongoing. The rare population of MSC with CFU-F capacity is found exclusively within the subpopulation of CD271+ cells that co-express CD73+/ CD105+/CD90+. MSC from MDS and AML show reduced frequencies compared to healthy samples and have a dramatically impaired CFU-F capacity. Preliminary gene expression analyses suggest a shift towards the adipogenic lineage in MDS-MSC, pointing to an imbalance between adipogenic and osteogenic differentiation as a contributing factor to disease progression in MDS. Disclosure: No conflict of interest disclosed.

Debatten

CLL -Zukunft ohne Chemotherapie ? V411

CLL – the future without chemotherapy? Buske C.1

ment-free remission in CML patients. To meet the challenges of the new era in CML treatment, the CML Alliance actively promotes innovation through cooperation not only between individual physicians but also between the various sectors involved in the diagnosis and treatment of CML. The four fields of activity are: Improving the clinical research infrastructure throughout Germany. The CML Alliance tutorial project works to improve CML patients’ access to treatment within a clinical trial by advising and supporting physicians and clinical personnel concerning all aspects of participating in clinical research – from information on specific studies to more general quality assurance issues. Information and network-building. Through symposia, GCP courses, and other informational seminars and workshops at both the local and national levels, the CML Alliance disseminates information on the latest developments in CML treatment, diagnostics, and research as well as the ins and outs of participation in clinical trials from both the physician and patient perspectives. Standardizing diagnostics requisite for the newest approaches to treating CML, i.e. therapy discontinuation. The CML Alliance supports the efforts to standardize quantitative BCR-ABL diagnostics according to the International Scale and the measurement of deep molecular response (MR4,5). Supporting patient advocacy. The patient perspective is an invaluable contribution to the efforts of physicians and researchers to optimize CML treatment. The CML Alliance supports the multifaceted activities of patients and patient advocates, including patient-information events and the development of patient-friendly information on clinical trials. The German CML Alliance is a project of the Jena University Hospital, currently funded through an unrestricted grant from Novartis. Disclosure: No conflict of interest disclosed. V415

Universitätsklinikum Ulm, CCC Ulm, Klinik für Innere Medizin III, Ulm, Germany

Innovative concepts for clinical trials in CML

In the recent years there has been dramatic progress in the development of new therapeutic approaches in chronic lymphocytic leukemia and other B – Non-Hodgkin´s lymphomas. Mostly phase II data have proven that these new compounds targeting e.g. signal transduction pathways or the anti-apoptotic protein BCL-2 are highly efficient and are able to induce remissions even in chemo-refractory disease. However, experience with these new drugs is still limited, many of them are given as a permanent treatment and costs of these drugs are substantial. Thus, the key question for the future will be, how to use these surely fascinating new compounds in an optimal way and whether to the end they are indeed able to replace chemotherapy, the backbone of our treatment in the last decades.

Hochhaus A.1, for the German CML Study Group and German CML Alliance

Universitätsklinikum Jena, Abt. Hämatologie/Onkologie, Jena, Germany

The German CML Alliance works to optimize the treatment of chronic myeloid leukemia (CML) in Germany. This expanding network of physicians, diagnosticians, researchers, and patient advocates was founded in 2014 to address the repercussions of the fact that, since the introduction of TKI therapy at the turn of the twenty-first century, the number of people living with CML has been steadily growing. The ever-increasing prevalence of CML has reoriented therapeutic and clinical research goals—most prominently, the achievement of treat-

The advent of tyrosine kinase inhibitors (TKIs) into the management of CML has profoundly improved patients’ prognosis. Survival of responders is approaching that of the general population. However, about 20% of patients do not respond or suffer from strong side effects. In several trials, TKI treatment has been stopped successfully in approximately half of the patients with deep molecular response. This has prompted the development of a new concept in the management of CML patients known as “treatment-free remission”. The future of CML treatment will be to define criteria for the safe and most promising discontinuation of TKI on one hand, and, on the other, to increase the number of patients available for such an attempt. To resolve this, new innovative studies targeting residual leukemia stem cells are underway. The major objectives of current studies are (i) improving efficacy and tolerability of available TKIs to achieve high rates of deep molecular response with good quality of life; (ii) enhancing efficacy for patients lacking optimal response using treatment combination targeting alternative signal transduction pathways (e.g., combined ABL/JAK inhibition, CoRNea trial); (iii) reducing TKI treatment duration, costs and side effects of therapy by identification of safe discontinuation strategies; (iv) evaluating immunotherapies (interferon alpha and checkpoint inhibitors) to activate T cell response against persisting stem cells (TIGER, ENDURE trials); (v) testing novel ways of BCR-ABL inhibition, e.g. by allosteric inhibition using the novel compound ABL001 as monotherapy or in combination with other TKIs. The optimal access to innovative trials will be achieved by recruitment of newly diagnosed CML patients into studies (e.g. TIGER, NiloDeepR, DasaHIT) or registries to guarantee timely treatment decisions based on standardized monitoring. Essential in CML management is guideline directed use of cytogenetic and molecular follow up with standardized methods to regularly assess the remission status. The coordinated man-

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–128

Disclosure: No conflict of interest disclosed.

Fortbildung

Innovation durch Kooperation – Die Deutsche CML-Allianz V414

Overview of the work of the German CML Alliance Kolb M.1 Deutsche CML-Allianz c/o Universitätsklinikum Jena, KIM II / Hämatologie und Intern. Onkologie, Jena, Germany 1

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agement between academic and regional hospitals, office-based hematologists, laboratories, and patient representatives within the German CML Alliance permits up-to-date patient care and the early use of novel therapeutic options in patients at risk. Disclosure: Andreas Hochhaus: Employment or Leadership Position: Universitätsklinikum Jena; Advisory Role: Novartis, BMS, Pfizer, Incyte (Ariad); Expert Testimony: Novartis, BMS, Pfizer, Incyte (Ariad) V416

Health care research in clinical practice Tesch H.1 Centrum für Hämatologie und Onkologie Bethanien, Frankfurt, Germany

There is a lack of data describing treatment management and results in patients with CML treated with TKIs or other treatments outside the setting of clinical trials. Assuming an incidence of CML of 1.0 -1.5/100,000/ year, there are about 1000 -1200 newly diagnosed CML- patients annually in Germany. However only about 25% of these patients will be recruited into clinical studies. Clinical trial populations are well defined and might differ from the general patient population in clinical practice. Therefore large prospective non-interventional trials aim to capture information on epidemiology, treatment practice and outcome. Optimal treatments strategies and guideline adherence is a major concern. Therapy management of the new oral therapies is challenging for patients as well as physicians. Counseling options for practice teams are available for patients to cope with daily life and oncological trained personnel are getting more important in supporting therapy management. Results of the ongoing studies will be reported. To get more insight in the real life situation of CML treatment in practice the “Arbeitskreis Klinische Studien” (AKS) initiated a number of non-interventional and community based research strategies to improve this situation. Results of the ongoing studies will be reported. In 2014 the AKS and the CML Cooperative Study Group initiated the German CML Alliance to build up a platform for all physicians and patients dealing with CML. The alliance has 4 major working groups ie diagnostics, clinical trials, quality control and education and will hopefully have a major impact for improving treatment results for CML patients. Disclosure: Hans Tesch: Employment or Leadership Position: Selbständiger niedergelassener Arzt; Advisory Role: Ja; Financing of Scientific Research: Ja V417

Use of tutors for the establishment of the study infrastructure in practice Chudziak D.1, Knorr P.2, Kolb M.2, Hochhaus A.3, Tesch H.4 Deutsche CML-Allianz, c/o Onco Medical Consult, Frankfurt, Germany, Deutsche CML-Allianz, c/o Universitätsklinikum Jena, Jena, Germany, 3 Universitätsklinikum Jena, Jena, Germany, 4Centrum für Hämatologie und Onkologie Bethanien, Frankfurt, Germany 1 2

Rationale: In Germany approximately 1,200 patients are newly diagnosed with Chronic Myeloid Leukemia (CML), of which only about 25% are treated in a clinical trial. Therefore, the German CML-Alliance was founded by the German CML Study Group and the AKS in October 2014 to build a platform for all physicians who treat CML patients and patients themselves. The aim of the association, which is currently funded by an unrestricted grant from Novartis, is to establish an optimal standard of care and to achieve treatment free remission in as many patients as possible. To achieve this, the CML Alliance promotes close cooperation between university hospitals, non-university hospitals, hematologists and oncologists in practice and patients´ representatives. The tutorial project:The tutorial project of the German CML Alliance represents an independent and innovative concept of the center and study support. The aim of the project is to help all interested parties to access clinical trials and to give everyone the opportunity to participate in innovative studies. Both Investigator Initiated Trials (IITs) and pharmaceutical

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studies are included.. Regional tutors visit interested hospital or practice centers and provide guidance about the conduct of studies. During these site visits the tutors check the suitability of sites for study participation (site requirements to participate in studies and potential patients) and support the centers in the compilation of documents and regulatory issues. During a study, physicians and staff have access to the experience of the tutors and get valuable assistance in the successful completion of studies. Through regional events also cooperation be promoted in order to facilitate cooperation of private physicians, hospitals and university institutions and increase the recruitment in phase I, phase II and phase III trials. Conclusions: By May 2016, more than 350 doctors have registered in the CML Alliance and agreed to cooperate. The tutors have visited and supported more than 88 centers across Germany and various training and education projects have been performed. All services performed by the tutors, are free of charge for the centers. Disclosure: Doreen Chudziak: No conflict of interest disclosed. Hans Tesch: Employment or Leadership Position: Niedergelassener Onkologe im Centrum für Hämatologie und Onkologie Bethanien Frankfurt.; Advisory Role: Beratungstätigkeit Novartis; Financing of Scientific Research: Novartis, BMS V418

Molecular monitoring 2016 Lange T.1 Asklepios Klinikum, Weißenfels, Germany

Molecular monitoring with quantitative RT-PCR is the standard for assessment of major molecular response (MMR, 0.1% IS BCR-ABL/ABL), one of the major goals of tyrosine kinase inhibitor (TKI) treatment in chronic myeloid leukemia (CML). For more than a decade, BCR-ABL levels normalized to the reference genes ABL or GUS have been standardized between different labs using conversion factors to adjust to the international scale (IS). In the meantime however, a substantial proportion of CML patients qualify for treatment discontinuation strategies if achieving deeper levels of major molecular response (MR) 4.0 (BCR-ABL/ABL 0.01% IS), MR4.5 (0,0032% IS) or MR5.0 (0.001% IS). Quantification at this level demands not just that results are comparable between laboratories but also that they are highly sensitive, with reliable detection of just one or two BCR-ABL copies in a PCR sample containing 107 white blood cells. Therefore, standardization to reliably detect MR4.5 is far more challenging than the laboratory-specific adjustment of results according to IS. Furthermore, rules used in the interpretation of results have recently been published und need to be taken into account. In the future, modern and more comparable PCR techniques such as digital PCR may play an increasingly important role in the monitoring of CML. Five labs are currently standardized for MR4.5 in Germany, with approximately ten more in the process of standardization. With this background of standardized labs, monitoring of CML for MR4.5 will be available for all patients. The CML Alliance accompanies the process of standardization in Germany together with the European Leukemia Network and the EUTOS III program. Training in interpreting BCR-ABL reports and help in solving logistical issues will also be provided. Disclosure: Thoralf Lange: Financing of Scientific Research: Novartis, BMS; Expert Testimony: Novartis

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CONTENTS AUTHOR INDEX

Fortbildung

Update Infektionen in der Hämatologie und Onkologie - 20 Jahre AGIHO V421

Community acquired respiratory virus (CRV) infections in cancer patients – guideline on diagnosis and management by the Infectious Diseases Working Party (AGIHO) of the German Society for Haematology and Medical Oncology (DGHO) von Lilienfeld-Toal M.1, AGIHO Klinik für Innere Medizin II, Universitätsklinikum Jena, Hämatologie und internistische Onkologie, Jena, Germany 1

Community acquired viruses (CRV) may cause severe disease in cancer patients. Thus, efforts should be made to diagnose CRV rapidly and manage CRV-infections accordingly. Methods: A panel of 18 clinicians from the Infectious Diseases Working Party (AGIHO) of the German Society for Haematology and Medical Oncology (DGHO) have convened to assess the available literature and provide recommendations on the management of CRV infections including influenza, respiratory syncytial virus (RSV), parainfluenza virus (PIV), human metapneumovirus (hMPV) and adenovirus. Results: CRV-infections in cancer patients may lead to pneumonia in approximately 30% of cases, with an associated mortality of around 25%. For diagnosis of a CRV-infection, combined nasal/throat swabs or washes/ aspirates give the best results and nucleic-acid amplifications techniques (NAT) should be used to detect the pathogen. Hand hygiene, contact isolation and face masks have been shown to be of benefit as general infection management. Causal treatment can be given for influenza, using a neuraminidase-inhibitor, and RSV, using ribavirin in addition to intravenous immunoglobulins. Ribavirin has also been used to treat PIV and hMPV, but data are inconclusive in this setting. Cidofovir is used to treat adenovirus pneumonitis. Conclusions: CRV-infections may pose a vital threat to patients with underlying malignancy. This guideline provides information on diagnosis and treatment to improve the outcome. Disclosure: Marie von Lilienfeld-Toal: Advisory Role: MSD, Celgene; Financing of Scientific Research: MSD, Celgene, Janssen Cilag, Gilead; Expert Testimony: MSD, Pfizer; Other Financial Relationships: Janssen Cilag, Celgene, Astellas, Gilead

tine, prednisone) which is curative in about two thirds of patients. Whether the IPI is suitable to identify subgroups requiring different treatments, remains controversial. Additional radiotherapy may have a role in early stages, bulky disease and manifestations persisting after chemotherapy. ABC lymphomas are characterized by activation of the NFkB pathway. Efforts to target this pathway have so far failed to improve outcome in affected patients. Lymphomas expressing MYC and/or BCL2 tend to respond less well to R-CHOP than other DLBCL. How to improve their outcome is the subject of ongoing research. Patients with chemosensitive DLBCL relapse are candidates for high-dose chemotherapy with autologous blood stem cell transplantation which induces long-term remissions in about half of them. Due to younger patient age and less widespread lymphoma manifestations, treatment results in PMBCL tend to be better than in DLBCL. Protocols other than R-CHOP have been advocated for its treatment, but there are no randomized trials demonstrating their superiority. BL is characterized by rapid tumor cell proliferation and a propensity to disseminate into the central nervous system. Retrospective studies suggest that protocols including hyperfractionated alkylating agents and high doses of methotrexate and cytarabine are superior to R-CHOP. If patients survive the phase of massive tumor expansion, outcome is excellent, particularly in young patients who tolerate treatment-related toxicity better than old patients. Disclosure: Ulrich Dührsen: Financing of Scientific Research: Amgen, Celgene, Janssen, Roche Pharma; Expert Testimony: Amgen, Celgene, Roche Pharma V432

Salvage therapy of diffuse large B cell lymphoma Witzens-Harig M.1 Universitätsklinik, Heidelberg, Germany

Recently, therapeutic options in relapsed diffuse large cell lymphoma have improved dramatically. This lecture will cover salvage chemotherapy, stem cell transplantation, targeted therapy and immunotherapy and promising combinations of these modalities. There will also be a special focus on different treatment approaches in medically fit versus unfit patients. Disclosure: No conflict of interest disclosed.

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Magenkarzinome

Fortbildung

Aggressive Non-Hodgkin-Lymphome V430

Aggressive B-cell lymphoma: State of the art

V434

First-line therapy for metastatic gastric cancer – what treatment for which patient? Lordick F.1 University Medicine Leipzig, University Cancer Center Leipzig (UCCL), Leipzig, Germany 1

Dührsen U.1 Universitätsklinikum Essen, Westdeutsches Tumorzentrum, Klinik für Hämatologie, Essen, Germany 1

Aggressive B-cell lymphomas comprise diffuse large B-cell lymphoma (DLBCL) and its variants and subtypes, primary mediastinal B-cell lymphoma (PMBCL), Burkitt lymphoma (BL) and a number of rare B-cell lymphoma entities which are generally treated like DLBCL. Based on gene expression, DLBCL is subdivided in germinal center B-cell (GCB) and activated B-cell-like (ABC) lymphomas. Apart from differences in clinical presentation, peculiarities in gene expression also justify separation of DLBCL from PMBCL which is more closely related to Hodgkin lymphoma. Positron emission tomography/computed tomography is the imaging modality of choice to define the extent of disease. Age, performance status, stage, extranodal involvement and lactate dehydrogenase activity define the International Prognostic Index (IPI) which is used to predict outcome. Irrespective of stage and IPI risk group, standard treatment of DLBCL is R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincris-

Abstracts

Randomized trials showed that cytotoxic treatment prolongs survival and provides improved symptom control in advanced gastric cancer. Combination chemotherapy (CTx) with cisplatin and 5-FU is a preferred 1st-line CTx in Europe, and oxaliplatin has shown equivalent efficacy compared with cisplatin. Oral fluoropyrimidines, i.e. S-1 and capecitabine, can substitute for 5-FU. Modern doublet regimens are preferred in the majority of patients on the basis of a balanced benefit-risk ratio. The combination of irinotecan and 5-FU is as effective as platinum-based doublets and can also be considered. But Irinotecan is not an approvedand registered drug in first-line treatment of advanced gastric cancer. Selected, fit and compliant patients, especially those with high tumor burden or potential secondary resectability, a third drug (mostly docetaxel) may be added as triple CTx led to higher responses rates and enhanced efficacy. Current study concepts evaluated the feasibility and efficacy of de-escalation protocols and maintenance treatment with less toxic drugs, e.g. S-1

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monotherapy. This concept is currently studied following response induced by more intensive induction CTx. The comprehension of tumor biology is increasing. In HER2 positive gastric cancer, the addition of the anti-HER2 monoclonal antibody trastuzumab to platin and FU has prolonged survival. Novel anti-HER2-directed drugs like pertuzumab are being explored. More genetic alterations have been defined in subgroups of gastric cancer, like MET or FGFR amplification. However, its role for therapeutic targeting remains to be elucidated. Anti-angiogenic treatment with ramucirumab has been established as an effective drug for second-line treatment of advanced gastric cancer. Its role for first-line is unclear even more as other anti-angiogenic drugs, especially bevacizumab, failed to show sufficient efficacy. In conclusion, combination CTx is a European 1st-line standard for treating advanced gastric cancer. Molecularly targeted agents are further explored, preferably on the backbone of chemotherapy doublets. Disclosure: Florian Lordick: Advisory Role: Amgen, Biontech, Boston Biomedical, Ganymed, Lilly, MSD, Nordic, Roche, Taiho; Financing of Scientific Research: Vortragshonorare mgen, Celgene, Roche, Lilly; Expert Testimony: Böhringer-Ingelheim, GSK, Fresenius Biotech,; Immaterial Conflict of Interests: Reisekostenunterstützung: Amgen, Bayer, MSD, Roche, Taiho V435

Second- and third-line therapy in gastric cancer / new options for therapy Schmalenberg H.1 Krankenhaus Dresden-Friedrichstadt, IV. Medizinischen Klinik, Dresden, Germany 1

For a long time second or further line treatment of metastatic or advanced gastric cancer was under discussion. After efficacy of conventional 2nd line chemotherapy with irinotecan (Thuss-Patience 2011) or docetaxel (Ford 2014) was shown in Phase III trials additional treatment options were given recently with the approved drug ramucirumab, a monoclonal antibody against VEGFR -2. Ramucirumab was superior to best supportive care (BSC) alone or in combination with paclitaxel to paclitaxel alone (Fuchs 2014, Wilke 2014). Different results were seen with lapatinib, showing insufficient activity in Her-2 amplified advanced gastric cancer (Lorenzen 2015). New targets are under investigation. For pembrolizumab, a monoclonal antibody against PD-L1, first promising data are available (Muro 2014). Everolimus showed no survival benefit in this setting (Ohtsu 2013) or is still under investigation in combination with paclitaxel in a german trial. AZD4547 is a selective FGFR inhibitor showing good response in gastric cancers with high level clonal FGFR2 amplification. A german trial with cabazitaxel in pretreated gastric cancer patients completed recruiting recently. Disclosure: Harald Schmalenberg: Employment or Leadership Position: Chefarzt IV. Medizinische Klinik, Krankenhaus Dresden-Friedrichstadt; Advisory Role: Advisory Board Lilly; Financing of Scientific Research: Novartis, Merck; Expert Testimony: Sanofi

Freier Vortrag

Graft-versus-Host-Disease V437

Vitamin B6 serum levels on day+100 after allogeneic stem cell transplantation predict incidence of severe chronic GVHD – influence of ATG and statins Schmidt K.1, Lehners N.1, Radujkovic A.1, Ho A.D.1, Dreger P.1, Luft T.1 Universitätsklinikum Heidelberg, Med. Klinik V, Heidelberg, Germany

Severe chronic graft versus host disease (cGVHD) is a serious complication after allogeneic stem cell transplantation (alloSCT). It associates with

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debilitating morbidity due to fibrotic and inflammatory changes in connective tissues mainly of skin, lungs, eyes and the gastrointestinal tract. Although alloreactive T lymphocytes are clearly involved in the induction of both mild and severe cGVHD, it is unpredictable which patients are prone to develop severe rather than mild chronic disease. Monokine induced by interferon gamma (MIG, CXCL9) has recently been correlated with cGVHD. Anthranilic acid (AA) reduced the severity of acute GVHD in mice. AA is the product of a pathway involving indoleamine 2,3-dioxygenase (IDO), tryptophane (Trp), kynurenine (Kyn), and Vitamin B6. Based on our observation that anti-thymocyte globulin (ATG) and statins independently reduce the incidence of severe cGVHD, we investigated if MIG, IDO, Trp, Kyn and Vitamin B6 serum levels are correlated with cGVHD. The incidence of cGVHD was evaluated in 554 patients who consented to participate in this observational study and who survived the first 6 months after alloSCT. Day +100 serum samples for measuring CXCL9/MIG, IDO, Trp and Kyn by ELISA were available for 350 patients and at onset of cGVHD for 185 patients. Furthermore, VitB6 was measured by HPLC on day +100 in 268 patients. Chronic GVHD occurred in 295 patients (54%), 98 (18%) of whom developed severe cGVHD, and 197 (36%) developed non-severe cGVHD. Our study shows that ATG and statins minimize severe chronic GVHD by distinct mechanisms. ATG associates with reduced immune activation markers (CXCL9/MIG) at cGVHD onset, but no changes in homeostatic serum markers on day+100 were observed. This is consistent with initial depletion of alloreactive T cells during the early post-transplant period and weaker immune activation after tapering immunosuppressive therapy. In contrast, statin intake induced IDO and reduced Trp serum levels on day+100. In parallel, low Kyn levels suggest that statins activate Kyn catabolism. Failure of statins to reduce Trp and Kyn levels on day 100 predicted higher probability of severe cGVHD. The possible contribution of Kyn degradation products is supported by our observation that Vitamin B6 levels strongly and independently predict lower incidence of severe cGVHD. The synergism of statin and Vitamin B6 is clinically relevant and warrants further studies. Disclosure: No conflict of interest disclosed. V438

The role of lymphangiogenesis and its inhibition during GVHD Mertlitz S.1, Shi Y.1, Kalupa M.1, Mengwasser J.1, Riesner K.1, Cordes S.1, Penack O.1 Charité – Universitätsmedizin Berlin (CVK), Berlin, Germany

The inhibition of lymphangiogenesis is an established therapeutic concept in oncology, which is currently tested in clinical trials. During allogenic tissue transplantation the inhibition of lymphangiogenesis attenuates graft rejection. On the other hand, disease severity increased after inhibition of lymphangiogenesis in models of skin inflammation and colitis. Despite its critical importance for tumor growth, allo-immune responses and inflammation, the role of lymphangiogenesis has not been investigated experimentally in HSCT. Methods: We used the murine acute GVHD models LP/J -> C57Bl/6 and C57Bl/6 -> BALB/c. Lymph vessel density was quantified by immunofluorescence staining with Lyve-1 and flow cytometry using CD31 and gp38 markers. To study the effect of inhibition of lymphangiogenesis on GVHD, we used the mF4-31c1 antibody, which blocks the binding of VEGF-C to the lymph vessel specific VEGF receptor 3. Further, we analysed the immune status of IgG and mF4 treated animals by FACS. Results: We fund significantly higher lymph vessel density in GVHD target organs as well as significant higher numbers of lymphatic endothelial cells in lymph nodes of allo-BMT vs. syn-BMT recipients (Fig.1A). In addition, we found that VEGF C was significantly up-regulated during GVHD on RNA level. Blocking of VEGFR3 inhibited GVHD-associated lymphangiogenesis in the colon and in lymph nodes (Fig.1B). Inhi-

Abstracts

CONTENTS AUTHOR INDEX

bition of lymphangiogenesis resulted in significant reduction of GVHD and improved survival (Fig.1C). In mF4-31c1 antibody treated allo-BMT recipients vs. control antibody treated allo-BMT recipients we found an increase in TH17 and TReg, as well as CD11b and CD11c positive cells in the spleen (Fig.1D). Further, the number of B cells was significant higher in lymph nodes of mF4-31c1 treated mice (Fig.1D). Anti-VEGFR3 treatment had no significant effects on hematopoetic engraftment. Conclusion: We present novel evidence that acute GVHD is associated to lymphangiogenesis in target organs and in lymph nodes. Our data shows that anti-VEGFR3 treatment ameliorates lethal GVHD and identifies the inhibition of lymphangiogenesis as a novel therapeutic strategy in the setting of HSCT.

in non-GVHD target organs (Fig.1B). Next, we found that VEGFR2 expression in liver was not different in allo vs. syn-BMT recipients (Fig.1C). Accordingly, VEGFR1+2 or VEGFA inhibition had no positive effects on GVHD (Fig.1D). We next looked at expression levels of endothelium-related genes and found up-regulation during established GVHD, i.a. activation marker MHCII (Fig.1E). In contrast, we found significant down-regulation of ICAM, VCAM, P-Selectin and vWF in allo vs. synBMT recipients in LEC at day+2 (Fig.1F) suggesting that alternative pathways may be critical for initiation of angiogenesis in GVHD target organs. We therefore performed the unbiased approach of proteomic profiling of LEC at day+2 (Fig.1G). Interestingly, the up-regulated proteins included several promising candidates that could be involved in initiation of angiogenesis such as CPT2, G6pdx, Bckdha, Marc2, Hspg2, Rdx, Ltf and Psmc2 (Fig.1H). Conclusion: Our data demonstrates that angiogenesis is a very early event during GVHD which is restricted to target organs. Our findings make it likely that angiogenesis is the initiating process of inflammation in GVHD target organs implicating that endothelial biology plays a major role in GVHD organ tropism and disease development. Our data suggests that non-classical pathways may be critical for initiation of angiogenesis in GVHD target organs and we identified candidates using a proteomic approach.

Fig. 1. Disclosure: No conflict of interest disclosed. V439

Angiogenesis in acute GVHD Riesner K.1, Shi Y.1, Kalupa M.1, McGearey A.1, Mengwasser J.1, Mertlitz S.1, Cordes S.1, Perez-Hernandez D.2, Dittmar G.2, Penack O.1 Charité – Universitätsmedizin Berlin (CVK), Klinik m.S. Hämatologie, Onkologie und Tumorimmunologie, Berlin, Germany, 2Max Delbrück Center for Molecular Medicine, Berlin, Germany 1

We asked the questions if angiogenesis is the initiating process of inflammation in GVHD target organs, if endothelial changes are specific to target organs and which were the molecular mechanisms. Methods: Murine acute GVHD models LP/J→B6, B6→Balb/c, B6→BDF and 129S2/Sv→B6. Immune staining, FACS and qPCR for CD31, CD4, CD8, CD11b, ICAM, VCAM, E/P-Selectin, vWF, MHCII, VEGFR2 and VEGFA. Treatment with DC101+MFI (Imclone), B20-4.1.1 (Genentech). Isolation of liver endothelial cells (LEC) by MACS and FLOW-sort. Proteome analysis based on mass spectrometry. Results: During GVHD we found significant angiogenesis in liver, colon and skin as early as day+2 (Fig.1A). Leukocyte infiltration occurred secondary to angiogenesis at day+7 or later. No angiogenesis occurred

Abstracts

Fig. 1. Disclosure: No conflict of interest disclosed.

Oncol Res Treat 2016;39(suppl 3):1–160

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V440

V441

No evidence of increased infection rates using low-dose alemtuzumab instead of ATG as GvHD prophylaxis before allogeneic stem cell transplantation

Improving the NIH cGVHD severity scores using salvage everolimus-based therapy in moderate and severe chronic GVHD

Neumann T.1, Schneidewind L.2, Pink D.3, Schulze M.4, Krüger W.1

Klink A.1, Hilgendorf I.1, Schilling K.1, Hochhaus A.1, Sayer H.G.1,2

Universitätsmedizin Greifswald, Klinik für Innere Medizin C (Hämatologie / Onkologie), Greifswald, Germany, 2Universitätsklinikum des Saarlandes, Institut für Virologie, Homburg (Saar), Germany, 3Helios-Klinikum Bad Saarow, Klinik für Hämatologie, Onkologie und Palliativmedizin, Bad Saarow, Germany, 4 Kreiskrankenhaus Wolgast, Klinik für Innere Medizin, Wolgast, Germany

Introduction: Alemtuzumab as part of the conditioning protocol is effective in reducing GvHD, but may be associated with increased infection rates, especially when using high doses (i.e. 100 mg over 5 days) Methods: We performed a retrospective, single-center, case-control study analyzing the rates of neutropenic fever, cytomegalovirus (CMV) reactivation, Epstein-Barr-virus (EBV) reactivation, clinical manifest toxoplasmosis, and human herpes virus 6 (HHV6) reactivation using low dose alemtuzumab in comparison with ATG as GvHD prophylaxis before allogeneic stem cell transplantation. 44 patients transplanted from unrelated donors during the period from 2001 to 2012 were analyzed. All patients were matched regarding age, diagnosis and conditioning regimen. One group of patients was treated with alemtuzumab (manufacturer Genzyme, USA) 10 mg at day -2 (respectively 20 mg in case of mismatch transplantation). The other group was treated with an alternative immunosuppressive regimen containing ATG and was either treated with ATG Fresenius (10 mg/kg for 3 consecutive days) or Thymoglobulin (2 mg/kg for 3 consecutive days). Only infections or reactivations requiring treatment were included in our analysis. Results: The results obtained in both groups were analyzed by the Wilcoxon rank-sum test. Rates of CMV reactivation, EBV reactivation, HHV6 reactivation and clinical manifest toxoplasmosis did not differ significantly between both groups (see Tab. 1) until 2 years after transplantation (end of observation period). No case of PTLD was observed in both groups. Also, rates of neutropenic fever during inpatient treatment after transplantation did not differ significantly in both groups. Both groups included 3 patients (14%) with a high risk of CMV reactivation (recipient CMV-IgG positive, donor CMV-IgG negative). Tab. 1. Infectious complications in both groups. Number of patients, percentage in parenthesis. Significance level p was calculated using Wilcoxon rank-sum test, n.s. means not significant.

Alemtuzumab ATG

No. of patients Neutropenic fever CMV reactivation EBV reactivation HHV6 reactivation Toxoplasmosis

22 12 (54%) 8 (36%) 1 (5%) 0 1 (5%)

22 13 (59%) 8 (36%) 0 1 (5%) 0

Significance Level (p) n.s. n.s. n.s. n.s. n.s.

Conclusion: We saw no evidence of increased infections rates when using low-dose alemtuzumab as GvHD prophylaxis before allogeneic stem cell transplantation in this retrospective analysis. Disclosure: No conflict of interest disclosed.

Universitätsklinikum Jena, Klinik für Innere Medizin II, Hämatologie und Onkologie, Jena, Germany, 2HELIOS Klinikum Erfurt, 4. Medizinische Klinik, Hämatologie, internistische Onkologie und Hämostaseologie, Erfurt, Germany

Introduction: The treatment of chronic graft-versus-host disease (cGVHD) is still a major challenge of allogeneic hematopoietic stem cell transplantation. The data available to other regimens including calicineurin inhibitors as second line treatment considering the National Institutes of Health (NIH) score are still limited. Here we report a retrospective analysis on the overall treatment response, organ-specific efficacy and safety of everolimus as salvage therapy in patients with moderate and severe cGVHD. Patient and methods: 38 patients (pts, 13 female, median age 47.9 years, range 28.9–65.6) were included. 18 patients (47%) suffered from moderate and 20 patients (53%) from severe cGVHD at start of treatment with everolimus. The median treatment time was 216 days (18–1418). All pts received everolimus as a combination therapy with prednisolone (PDN, n = 17) or PDN and Mycofenolate Mofetil (MMF, n = 16) or PDN and MMF and extracoporal photophoresis (n = 5). Results: Twenty pts (52.6%) achieved an improvement in the NIH global score. Regarding the degree of remission 4 pts (10.5%) reached a complete remission (CR), 13 pts (34.2%) a partial remission (PR), 11 pts (28.9%) had a mixed response (MR), 6 pts (15.8%) a stable disease (SD) and 4 pts (10.5%) a progressive disease (PD). Median time to PR was 3 months (range 1–9) and to CR was 23 months (range 2–39). Number of treatment line (2nd to 5th) and the concomitant immunosuppressant medication had no significant effect on the response to treatment. In 20 pts (52.6%) PDN dose was reduced to a median of 0.12 mg/kg daily (0.03 - 0.38) and 11 pts (28.9%) achieved a CR or PR (p = 0.046). Analysis of the organ-related response with CR and PR showed the following results: 21/33 pts (63.6%) with cutaneous cGVHD, 17/26 pts (65.4%) with oral mucosa cGVHD, 9/30 pts (30.0%) with ophthalmic cGVHD, 2/6 pts (33.3%) with gastrointestinal cGVHD, 9/20 pts (45.0%) with liver cGVHD, 3/10 pts (30.0%) with lung cGVHD, 0/2 pts with genital cGVHD and 3/5 pts (60.0%) with fascial and muscular cGVHD. Serious grade 3/4 toxicities were observed as arterial embolism (n = 1), hypertriglyceridemia (n= 4), cholecystitis (n = 1), myalgia (n = 3), cutaneous tumor (n = 1) and infectious diseases (n = 5). Conclusion: Everolimus as salvage line treatment achieves in cGVHD response even in higher treatment line (≥ 2nd) measured by the NIH cGVHD severity score. Studies that randomized everolimus against other salvage-line therapies are indicated Disclosure: No conflict of interest disclosed. V442

Serotherapy with anti-T cell globulins (ATG) may improve survival in allogeneic blood stem cell transplants applying methotrexate (MTX)-free graft-versus-host disease (GVHD) prophylaxis Clausen J.1, Böhm A.1, Strassl I.1, Stiefel O.1, Binder M.1, Buxhofer-Ausch V.1, Machherndl-Spandl S.1, Rotter N.1, Lechner D.1, Schmid S.2, König J.1, Weltermann A.1, Nachbaur D.2 Elisabethinen-Krankenhaus Linz, Interne I (Hämatologie & Onkologie), Linz, Austria, 2Universitätsklinik für Innere Medizin, Hämatologie und Onkologie, Innsbruck, Austria 1

Introduction: ATG is used to decrease the risk for severe acute and chronic GVHD following allogeneic hematopoietic stem cell transplantation (HSCT). Supporting evidence from randomized studies is largely based on myeloablative (MAC) transplants incorporating MTX in addition to a calcineurin inhibitor (CNI) for GvHD prophylaxis. On the other

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CONTENTS AUTHOR INDEX

hand, significant benefits of ATG in reduced-intensity conditioning (RIC) based HSCT, typically substituting mycophenolate mofetil (MMF) for MTX, are well documented in retrospective studies. However, the effect of ATG on overall survival (OS) has not been systematically compared in the two above scenarios. Methods: For this retrospective two-center analysis, we evaluated 505 related or unrelated, at least 10/10 HLA-matched peripheral blood HSCT, for adults with hematological diseases (185 with low-risk and 320 with intermediate- or high-risk disease), consecutively performed at two Austrian HSCT centers. Median follow-up of survivors is 43 months. Among RIC transplants (n = 176), the majority (n = 153) included MTX-free (i.e. mainly MMF-based) GVHD prophylaxis, and ATG was added in 86 HSCT. In MAC HSCT (n = 329), GVHD prophylaxis was predominantly MTX-based (n = 201), and ATG was added in 114 HSCT. Serotherapy was performed with either ATG-Fresenius/Neovii (n = 158; median dose 30mg/kg), Thymoglobulin (n = 36, median dose 5mg/kg), or Alemtuzumab (n = 6, 50–100 mg flat). Results: In MTX-free transplants (n = 281), 5-year OS was significantly better if ATG serotherapy was used to augment pharmacological GVHD prophylaxis (40% vs 26%, p = 0.038). By Cox multivariate analysis adjusting for confounding factors, the effect of ATG on OS remained significant (relative risk/RR, 0.60; p = 0.024). By contrast, in MTX-based HSCT (n = 224), serotherapy with ATG provided no significant survival benefit. In RIC transplants, 5y-OS was superior if ATG was applied (45% vs 24%, p = 0.037), although this effect did not reach significance in the multivariate analysis (adjusted RR, 0.71; p = 0.10). In MTX-free MAC transplants (n = 128), there was a pronounced survival benefit for ATG (adjusted RR, 0.52; p = 0.039). Conclusion: Together, our data suggest that an MTX-free GVHD-prophylactic regimen, irrespective of the conditioning intensity, is associated with a positive survival effect of ATG. Augmentation of GVHD prophylaxis through ATG may be helpful by compensating for the relatively low efficacy of an MTX-free regime.

Due to the recurrence of thrombotic events in some patients treated by low dose aspirin, questions have been raised regarding the existence of aspirin resistance. The so-called „turnover“ resistance has prompted some authors to recommend a bi-daily aspirin administration. Whether this biological resistance translates into increased thromboembolic risk has not been clearly demonstrated. The optimal duration of treatment with vitamin K antagonists (VKA) after venous thrombosis in patients with MPN is uncertain. Recurrent thrombosis is significantly reduced by VKA; however, the incidence of recurrence on treatment remains high. MPNs are the leadings systemic cause of non-cirrhotic splanchnic vein thrombosis (SVT). Cytoreduction is warranted in all SVT patients with an overt MPN, but its appropriateness is doubtful in those with molecular MPN without hypercythaemia. JAK2V617F mutation is the major contributory factor for the pathogenesis of thrombosis in MPN. The International Prognostic Score for thrombosis in ET (IPSET-thrombosis) identified JAK2 mutation as an independent risk factor. So MPNs could be ideal candidates for personalized medicine. The treatment of patients with MPN during pregnancy is a challenge in clinical practice. In this seminar, the relevant diagnostic and therapeutic aspects of the MPN associated thrombosis will be illustrated using examples from patients which we have taken care of and hopefully interesting cases from the auditorium will be discussed.

Disclosure: Johannes Clausen: Financing of Scientific Research: Berater-Honorar: Neovii Biotec David Nachbaur: No conflict of interest disclosed.

Disclosure: No conflict of interest disclosed.

Fortbildung

Management myeloproliferativer Neoplasien V451

Management of therapy in ET, PV and MF Griesshammer M.1 Johannes Wesling Klinikum Minden, Universität Bochum, Minden, Germany

Myelofibrosis (MF), polycythemia vera (PV) and essential thrombocythaemia (ET) are BCR-ABL1 negative MPNs. The MF is dominated by the effects of progressive bone marrow fibrosis resulting in shortened survival. Elevated thrombosis risk is the primary clinical concern in PV and ET. Risk stratification of patients with PV/ET by age and/or prior thrombosis provides the basis of risk adapted therapy. Aggressive control of cardiovascular risk factors, the use of aspirin, control of the hematocrit less than 45% in PV, and cytoreductive therapy in high-risk PV/ET is the focus of management. Hydroxyurea is the most common first-line therapy. The current evaluation of IFN-alpha in randomized study will provide clarity to the potential long-term benefit and risks associated with this biologic in PV/ET. Given the expanding role of targeted agents like ruxolitinib within the MPN arena, it will be of key importance to delineate the effects of these agents upon thrombotic risk.

Before planning therapy in MPN a careful medical history should be taken with special attention to MPN relevant symptoms, current medications, family history and cardiovascular risk factors. This must be followed by a clincal examination and sonography of the abdomen. Therapy in MPN is stratified according to a risk stratification based on quite simple clinical and laboratory data. In ET and PV risk stratification is based on age and prevoius thromboembolic events. In ET a platelet count > 1500 G/l is a risk factor for bleeding and an argument for cytoreduction. For older ET and PV patients hydroxyurea is the standard of therapy for cytoreduction, for younger patients, although not liscensed, IFN is a good option in PV and anagrelide in ET. In PV, phlebotomy and low dose aspirin are the cornerstone of therapy. In ET, aspirin has a place in patients with microcirculatory disturbances, cardiovascular events or older age. In PV patients resistant or intolerant to hydroxyurea ruxolitinib is the effective and liscensed drug of choice. Low risk ET patients should not be treated. They do have an excellent prognosis. Myelofibrosis patients (MF) are stratified according to the IWG-MRT risk scoring system into 4 risk groups according to age, leukocyte number, hemoglobin level, peripheral blasts and constitutional symptoms. High and intermediate 2 risk patients < 70 years are potential candidates for allogeneic perpheral stem cell transplantation. In MF with splenomegaly and/or constitutional symptoms ruxolitinib is the effective and liscensed drug of choice. Ruxolitinib is also used successfully in MF prior to transplantation or in severe graft versus host disease. A difficult clinical problem is the treatment of anemia in MF herein erytrhopoietins or androgens may be used successfully in some cases. A prerequisite for erytrhopoietin use in MF and anemia is an Epo level < 120 U/ml otherwise the choice should go for androgens to effectively treat anemia. Although not liscensed there are very interesting and disease modifying data for IFN in early MF. In all MPNs there is a great need for new therapies including new Jak inhibitors and other innovative new drugs.

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–160

Expertenseminar

Diagnostische und therapeutische Fallstricke bei MPNassoziierter Thrombophilie V444

Diagnostic and therapeutic pitfalls of the BCR-ABL1 negative myeloproliferative neoplasms (MPNs) associated thromboembolism Schilling K.1 Universitätsklinikum Jena, Klinik für Innere Medizin II, Jena, Germany

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Disclosure: Martin Griesshammer: Advisory Role: Baxalta, Novartis, Shire, AOP Orphan; Financing of Scientific Research: Baxalta, Novartis, Shire, AOP Orphan, Jansen V453

Stem cell transplantation in myelofibrosis Kröger N.1 Klinik für Stammzelltransplantation, Hamburg, Germany

Primary or post ET/PV myelofibrosis is one of the Philadelphia- negative myeloproliferative neoplasms with worst survival. Allogeneic stem cell transplantation (ASCT) can cure a substantial number of patients but is still not universally applicable due to toxicity which leads to therapy-related morbidity and mortality. In the more recent years outcome of ASCT has improved by less toxic conditioning regimens and optimization of relapse prevention strategies. Only two prospective studies with a larger sample size are available so far. The European Group of Blood and Marrow Transplantation (EBMT) published results of a prospective trial including 103 patients who received a busulfan/fludarabine-based reduced intensity conditioning regimen followed by related or unrelated stem cell transplantation. The median age was 55 years and the NRM at 1 year was 16%. Cumulative incidence of relapse was 22% at 3 years. Progression-free(PFS) and overall survival (OS) at 5 years were 51% and 67%, respectively. Advanced age and HLA-mismatched donor were independent predictive factors for reduced OS The Myeloproliferative Disorders Research Consortium performed a prospective phase II trial including 66 patients with primary or post ET/PV myelofibrosis investigating a reduced conditioning regimen with melphalan and fludarabine. With a median follow-up of 25 months overall survival was 75% in the sibling group and only 32% in the unrelated group due to a higher non-relapse mortality in the unrelated donor group (59% vs. 22%). The introduction of novel therapies such as JAK2 inhibitors may also be helpful in preparation of the transplant by reducing spleen size and constitutional symptoms. To reduce the risk of relapse molecular monitoring and adoptive immunotherapy with donor lymphocytes have been introduced. Despite lacking prospective randomized trials it is justified to offer ASCT to eligible patients with PMF whose median survival is expected to be less than 5 years. This includes patients with intermediate-2 and high risk according to IPSS or DIPSS, respectively. The benefit/risk ratio should be considered in each patient taking also transplant- and patient-specific factors into account. Disclosure: Nicolaus Kröger: Financing of Scientific Research: Novartis, Shire, Neovii, Jazz, Riemser, Medac; Expert Testimony: Novartis

Fortbildung

Management des Hodgkin-Lymphoms V454

Treatment of elderly Hodgkin Lymphoma patients Böll B.1 Uniklinik Köln, Centrum für Integrierte Onkologie, Innere Medizin I, Deutsche Hodgkin Studiengruppe, Köln, Germany 1

Introduction: About one in four patients with first diagnosis of Hodgkin Lymphoma (HL) is 60 years old or older. Importantly, the incidence of elderly HL due to the demographic changes in developed countries is steadily rising. Data on elderly HL patients is limited to few prospective trials and current treatment standards are mostly derived either from population-based studies or retrospective analyses or even indirectly derived from studies on younger HL patients. Methods: This presentation summarizes current knowledge and recent findings in the treatment of HL in elderly. In addition, currently recruiting trials in elderly HL implementing novel immunotherapeutic strategies are presented.

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Results: The overall prognosis for older HL patients is poor in spite of all advances achieved in younger HL patients. This is particularly true for elderly advanced stage patients, as excessive toxicity impedes effective treatment and the optimal first-line treatment is not known in these patients. ABVD has long been considered standard of care in elderly HL patients and might still be appropriate in early stage patients with some exceptions. However, in advanced stage patients we and others have noted severe toxicity with ABVD due to cumulative effects of 6–8 cycles. Similarly, alternative regimens as BACOPP or the gemcitabine-based PVAG have resulted in inacceptable toxicity and still lacked efficacy. Recently, we concluded a phase I study using lenalidomide combined with a truncated ABVD-regimen (AVD) and reported high efficacy including novel non-genotoxic substances into the first-line in older patients. Similarly, the anti-CD30 immunotoxin brentuximab vedotin is being used for elderly HL patients as monotherapy and in different combinations. Finally, checkpoint inhibitors targeting the PD1-pathwas Nivolumab and Pemrolizumab have been implementes for relapsed refractory patients with impressive results and are now being implemented in the treatment of elderly HL patients, too. Conclusions: Recent advances in the treatment of older HL patients might allow not only a more effective treatment in the first line but also a more individualized approach in case of relapse or progression. This presentation will give a comprehensive overview on current standards and will also report on currently recruiting trials in elderly HL patients. In addition, the current strategies and future approaches within the GHSG and beyond will be presented. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium

Therapeutische Targets bei AML – Vision und Wirklichkeit V458

Isocitrate dehydrogenase mutations in AML Krämer A.1 Klinische Kooperationseinheit Molekulare Hämatologie/Onkologie, Medizinische Klinik V, Universität Heidelberg und Deutsches Krebsforschungszentrum, Heidelberg, Germany 1

Isocitrate dehydrogenase (IDH) 1 and 2 are key metabolic enzymes, converting isocitrate to α-ketoglutarate. Active site, somatic mutations in IDH1 and IDH2 are associated with neomorphic enzyme activity, leading to the production of high levels of the oncometabolite 2-hydroxyglutarate (2-HG), and have been identified in multiple human cancers, including glioma, sarcoma, myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML). Mutations in IDH1 and IDH2 are mutually exclusive in AML and promote a block in cellular differentiation and progression to leukemia due to global DNA and histone hypermethylation, which occurs through accumulation of 2-HG and subsequent inhibition of histone and DNA demethylases as well as TET2 function. Mutations in IDH1 and IDH2 are found in 5–10% and 10–15% of AML cases, respectively. Inhibitors of both mutant IDH1 and IDH2 are currently in clinical trials and early results have shown encouraging results with remarkable activity and manageable toxicity in patients with relapsed/refractory AML. Mechanistically, the activity of IDH inhibitory compounds seems to be based on the induction of differentiation of malignant cells. Interim results of a phase I/II study of AG-221, the first IDH2 inhibitor in clinical trials demonstrated an overall response rate of about 40% and a CR rate of 18% in patients with relapsed/refractory AML. Early results for inhibitors of mutant IDH1 have shown evidence for efficacy similar to that of IDH2 inihibition. Thus, inhibition of mutant IDH shows promise as a treatment approach in AML, with further development ongoing in solid tumors and glioma. Disclosure: Alwin Krämer: Expert Testimony: Bayer

Abstracts

CONTENTS AUTHOR INDEX

V459

Exploring and exploiting chromatin dependencies in leukemia

Fortbildung

Supportive Therapien: aktuelle Leitlinien in der onkologischen Praxis

Roth M.1, Rathert P.1, Zuber J.1 Institute of Molecular Pathology (IMP), Wien, Austria

The genetic complexity and heterogeneity of human leukemias pose a daunting challenge for the development of effective targeted therapies. Despite their diversity, individual mutations converge at the functional level to dysregulate basic cellular processes, which is thought to result in exploitable dependencies (“non-oncogene addictions”). To systematically explore such dependencies, our lab combines genetically engineered mouse models (GEMMs) and a miRNA-based RNAi optimized for multiplexed screening and studies in vivo. A major focus of ongoing screens is on deciphering dependencies on chromatin regulators, which are required for maintaining aberrant cellfate programs in leukemia. To explore this promising target space, we have constructed an shRNA library targeting 700 chromatin regulators, which we have used to establish chromatin dependency profiles in a variety of leukemia models involving different driver mutations and tissue contexts. To investigate how the tissue of origin influences these dependencies, we have generated and screened a panel of isogenic cancer models driven by endogenous NrasG12D, loss of p53 and overexpression of Myc. Results of comparative screens reveal that, (1) Myc/NrasG12D/p53null cancers develop common as well as tissue-restricted chromatin dependencies, which are particularly prominent in hematopoietic cancer; (2) suppression of several chromatin regulators has opposing effects in isogenic cancers; and (3) as one of the strongest yet tissue-restricted effects we identify Brd4 as dependency in myeloid and B-cell leukemia, suggesting that the broad activity of BET inhibitors in these malignancies is mainly determined by tissue context. While several BET inhibitors have already entered clinical phase-I trials, the mechanisms underlying sensitivity and resistance to these agents remains poorly understood. Using dynamic drug response profiling and focused RNAi-based studies and screens, we have identified epigenetic mechanisms underlying primary and acquired resistance to BET inhibition. Our studies reveal that myeloid leukemias can develop resistance to BET inhibition through rewiring the transcriptional regulation of key BRD4-dependent target genes such as MYC. This process involves specific changes in the enhancer landscape and the activation and recruitment of WNT signaling machinery, which drives resistance in leukemia models and may serve as a biomarker for predicting BET resistance in the clinic Disclosure: No conflict of interest disclosed. V460

Normal and malignant stem cells and their niches Krause D.S.1 Georg-Speyer-Haus und Goethe Universität Frankfurt, Frankfurt am Main, Germany 1

This talk will focus on the concept and function of the stem cell niche as first described by R. Schofield in 1978. The nature and physiology of a niche, using the example of the normal haematopoietic stem cell niche, as well as recent findings about this niche will be discussed and juxtaposed to how cancer stem cells reciprocally interact with their specific niche or tumour microenvironment by employing the example of leukaemia. The talk will elaborate on how chemoresistance may be mediated by the niche and how the niche can potentially be targeted in future. Disclosure: Daniela Krause: Advisory Role: Glycomimetics Inc., USA

Abstracts

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Antiemetic recommendations of the German evidence based guideline „Supportive therapy for patients with cancer”. Focus on the multidisciplinary setting and different patient Groups. Jahn F.1 Universitaetsklinik Halle/Saale, Hämatologie /Onkologie, Halle, Germany

Introduction: Side effects associated by cancer therapy are still dreaded by patients and therapists. Therefore it is high priority to implement high quality, evidence based S3 guidelines, considering the improved therapeutic options and resent research successes. The aim is to increase the patients quality of life in the daily clinical practice. Methods: The German evidence based S3 guideline is mandated by ASORS, DGHO and DEGRO, Prof. Karin Jordan from the University Hospital Halle/ Saale is the guideline coordinator in conjunction with Dr. Franziska Jahn. Financial and scientific support was given by the German guideline program in Oncology, an initiative of the Association of Medical Scientific Societies (AWMF), the German Cancer Society (DKG) and the German cancer Aid (DKH). 76 Members and experts from 42 different professional societies and organizations, including Austria and Switzerland, developed recommendation of the 10 topics of supportive care chosen for the first edition of the guideline: Anemia, neutropenia, neurotoxicity, diarrhea, mucositis, scin toxicities, bone complications, chemotherapy induced emesis and nausea, paravasation and supportive care in radiotherapy. Predominantly we performed systematic literature searches based on key questions. Evidence based recommendations have been developed, discussed in task groups and shaped and adjusted in the panel meetings. Results: The chapter about nausea and vomiting during cancer therapy is based on both, the adaption of the ASCO antiemetic guidelines and a systematic literature search to include all relevant current study results. Among the recommendations for the antiemetic prophylaxis, the guideline defines the levels of emetogenicity, information about the available drugs, potential risk factors and therapeutic strategies, if the prior antiemetic prophylaxis turned out to be insufficient. Furthermore radiotherapy induced nausea and vomiting was also involved in the guideline process. In the oral presentation different patient groups and there particular needs are discussed as well. Disclosure: Franziska Jahn: Financing of Scientific Research: MSD, Riemser, Tesaro; Other Financial Relationships: Amgen V463

Drug interactions (DI) during supportive care Lipp H.-P.1 Universitätsklinik Tübingen, Universitätsapotheke, Tübingen, Germany

Avoidance of clinically relevant drug interactions (DI) during treatment with anticancer drugs incl. cytotoxic agents (CTX) as well as targeted therapeutics is a key challenge in clinical oncology. In this context, dramatic outcomes have been described with mercaptopurines and xanthine oxidase inhibitors or fluoropyrimidines and brivudine. However, within the last months, important DI have been desribed during use of PPI and selected TKI as well as Capecitabine with impaired drug solubilization in gastric pH as underlying mechanism. If a change to another TKI is not feasible, a concomitant intake of Coca Cola or Betain-HCl may be warranted. In case of Cyp450-dependent DI, the NK1 antagonists Aprepitant and Netupitant will increase absolute bioavailability of drugs which undergo an extensive Cyp3A-mediated first pass effect in the gut (e.g. Quetiapine, Sirolimus), in contrast to Rolapitant, which, however, imapirs Cyp2D6-related metabolism (e.g. Thioridazine). In addition, several co-analgetic

Oncol Res Treat 2016;39(suppl 3):1–160

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agents may be resposnsible for considerable Cyp450-induction (e.g. CBZ, OxCBZ, Topiramate) or Cyp2D6 inhibition (e.g. selected SSRI). In case of herb-based CAM-DI (CAM: complementary and alternative medicine), anticancer drug pharmacokinetics may be significantly altered, as it has been impressively shown for St. John´s wort (e.g. SN38, Imatinib) or Green Tea Extracts (e.g. Bortezomib, Sunitinib, Tacrolimus), however, the underlying mechanisms for the latter have not yet been elucidated in detail. Most often, experience to supportive care-related DI is primarily revealed by case reports or series, however, conflicting results may be available (e.g. ondansetron and CTX) which is often related to small patient numbers and insufficent randomisation, which highlights the need for cautious analysis of PubMed data. As a consequence, more extensive data collection and critical interpretation of those data regarding DI in clinical oncology is mandatory. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Immuntherapie von Leukämien und Lymphomen

analysis: 7/7 patients showed responses to CMVpp65 and 2/7 exhibited responses to PRAME and WT1, respectively. DC-activated T cells showed an upregulation of PD-1 and LAG-3 in vitro. Therefore, we studied the potential of checkpoint blockade to further enhance T-cell activation by DCs. We found that both pathways target different T-cell subsets, as PD-1 blockade resulted in increased IFN-γ secretion by TN- and TEM-subsets, while blockade of LAG-3 significantly affected TN- and TCM-subsets. Conclusion: We conclude that vaccination with next-generation LAA-expressing DCs in AML is feasible, safe, and induces anti-leukemic immune responses in vivo. Our in vitro data supports the hypothesis that the vaccine effects could be further enhanced by PD-1 and LAG-3 blockade. Disclosure: No conflict of interest disclosed. V465

Induction of NK cell reactivity against myeloid leukemia by a novel Fc-optimized CD133 antibody Schmied B.J.1, Andre M.2,3, Körner S.1, Leibold J.4, Bühring H.-J.5, Haen S.5, Kübler A.2,3, Kanz L.5, Grosse-Hovest L.6, Jung G.4, Salih H.R.1,2 Clinical Collaboration Unit Translational Immunology, German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Partner site Tübingen, Department of Hematology and Oncology, Tübingen, Germany, 2 University Children´s Hospital, Department of Pediatric Hematology and Oncology, Eberhard Karls University, Tübingen, Germany, 3University Children´s Hospital, Department of Pediatric Intensive Care, Basel, Switzerland, 4 Department of Immunology, Eberhard Karls University, Tübingen, Germany, 5 Department of Hematology and Oncology, Eberhard Karls University, Tübingen, Germany, 6Synimmune GmbH, Tübingen, Germany 1

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Induction of antigen-specific T-cell responses through dendritic cell vaccination in AML: results of a phase I/II trial and ex vivo enhancement by checkpoint blockade Lichtenegger F.S.1,2,3, Deiser K.1,2,3, Rothe M.1,2,3, Krupka C.1,2,3, Schnorfeil F.M.1,2,3, Augsberger C.1,2,3, Köhnke T.1,3, Bücklein V.1,3, Altmann T.1,3, Moosmann A.4, Brüggemann M.5, Heemskerk M.H.M.6, Wagner B.7, Hiddemann W.1,3, Bigalke I.8, Kvalheim G.8, Subklewe M.1,2,3 Klinikum der Universität München, LMU Munich, Department of Internal Medicine III, München, Germany, 2Helmholtz Zentrum München, Clinical Cooperation Group Immunotherapy, München, Germany, 3German Cancer Research Center (DKFZ), German Cancer Consortium (DKTK), Heidelberg, Germany, 4Helmholtz Zentrum München, DZIF Research Group Host control of viral latency and reactivation, München, Germany, 5University Hospital Schleswig-Holstein, Department of Hematology, Kiel, Germany, 6Leiden University Medical Center, Department of Hematology, Leiden, Netherlands, 7 Klinikum der Universität München, LMU Munich, Department of Transfusion Medicine, Cellular Therapeutics and Hemostaseology, München, Germany, 8The Norwegian Radium Hospital, Oslo University Hospital, Department of Cellular Therapy, Oslo, Norway 1

Introduction: Postremission therapy for acute myeloid leukemia (AML) is critical for elimination of minimal residual disease (MRD). Vaccination with autologous dendritic cells (DCs) loaded with leukemia-associated antigens (LAAs) is a promising strategy to induce anti-leukemic immune responses. Methods: We conduct a phase I/II proof-of-concept study using monocyte-derived next-generation DCs as postremission therapy of AML patients with a non-favorable risk profile in CR/CRi after intensive induction therapy (NCT01734304). These DCs are generated using a GMP-compliant 3-day protocol including a TLR7/8 agonist, loaded with RNA encoding the LAAs WT1 and PRAME as well as CMVpp65 as adjuvant/surrogate antigen, and are applied i.d. up to 10 times within 26 weeks. The primary endpoint of the trial is feasibility and safety of the vaccination. Secondary endpoints are immunological responses and disease control. Results: 12 patients have been enrolled into the trial. Phase I was completed successfully (n = 6), and phase II has been initiated. DCs of sufficient number and quality were generated from leukapheresis in 10/11 cases. DCs exhibited an immune-stimulatory profile based on high costimulatory molecule expression, IL-12p70 secretion, migration towards a chemokine gradient and processing and presentation of antigen. In 9/9 vaccinated patients, we observed delayed-type hypersensitivity (DTH) responses at the vaccination site, but no grade III/IV toxicities. TCR repertoire analysis by next-generation sequencing revealed an enrichment of particular clonotypes at DTH sites. In the peripheral blood, we detected vaccination-specific T cell responses by multimer staining and by ELISPOT

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NK cells largely contribute to the success of monoclonal antitumor antibodies (mAbs) in cancer due to their ability to mediate antibody-dependent cellular cytotoxicity (ADCC), a feature widely considered critical for therapeutic success. Up to now, no immunotherapeutic antibodies are available for the treatment of myeloid leukemias. Recently, we reported on the development of mAbs targeting CD133, which is expressed on a wide variety of tumor cells (Rothfelder et al., Blood 2015 126:3793). Here we extend our analyses and provide further data on the preclinical characterization of an Fc-engineered CD133 mAb developed for the treatment of myeloid leukemias. The mAb clone 293C3 was found to recognize leukemic cells in a higher percentage of patients with AML and CML than two other available mAb (clones AC133 and W6B3). Accordingly, clone 293C3 was chosen to generate chimeric mAb with either a wildtype Fc part (293C3-WT) or a variant containing amino acid exchanges (S239D/ I332E) to enhance affinity to the activating Fc receptor CD16 on NK cells (293C3-SDIE) that then underwent preclinical characterization. As mAbs can induce a pronounced shift of their target antigen which can largely limit their therapeutic efficacy, we studied CD133 levels on leukemia cells upon exposure to our mAb. 293C3-SDIE did not reduce expression below 50%, even after 72h, which should allow for induction of potent ADCC upon therapeutic application. While 293C3-SDIE was found not to influence complement-dependent cytotoxicity, treatment significantly induced activation, degranulation and lysis of primary CD133-positive AML cells by NK cells in allogeneic and autologous experimental in vitro models. Neither relevant toxicity of 293C3-SDIE at the level of committed hematopoietic progenitor cells nor induction of lysis by allogeneic or autologous NK cells was observed with healthy bone marrow (BM) cells that reportedly can express CD133. In a NOD.Cg-Prkdcscid IL2rgtmWjl/Sz (NSG) xenotransplantation model, treatment with 293C3-SDIE resulted in the elimination of patient AML cells due to induction of ADCC by NK cells from a matched human donor. Thus, in our view 293C3-SDIE constitutes an attractive immunotherapeutic compound particularly suitable for elimination of minimal residual disease in CD133 bearing leukemia in the context of allogenic SCT. Disclosure: No conflict of interest disclosed.

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V466

T cell stimulation with different cytokines results in distinct phenotypes and cytotoxic activity of CD19-specific CAR T cells Hoffmann J.-M.1, Gern U.1, Wang L.1, Kleist C.2, Wenthe J.3, Hückelhoven A.1, Schmitt A.1, Wuchter P.1, Sellner L.1, Schubert M.-L.1, Yoo H.1, Dreger P.1, Ho A.D.1, Loskog A.3, Brenner M.K.4, Schmitt M.1 Medizinische Klinik V, Universitätsklinikum Heidelberg, Heidelberg, Germany, Nuklearmedizin, Universitätsklinikum Heidelberg, Heidelberg, Germany, 3 Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden, 4Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children´s Hospital and Houston Medthodist Hospital, Houston, United States 1 2

Introduction: CAR T cell (CART) therapy became a promising option for the treatment of relapsed or refractory leukemia/lymphoma patients. However, efficiency of CART cells for a specific patient is fairly heterogeneous and not yet fully understood. The outcome might be related to the phenotype of the CART, i.e. naive (N), central / effector memory (CM/ EM), effector (E) cells, and related to the cytokine cocktail used for CART culture. Freezing/thawing might also have an impact on number, phenotype and function of CART cells. Methods: CART from the same donor were generated by a CD19. CAR-CD28-CD137zeta vector (M. Brenner) and two different cytokine cocktails: IL-7/IL-15 plus anti-CD3/anti-CD28 antibodies vs. IL-2 plus anti-CD3 antibody. Cytotoxicity was assessed by a standard chromium (Cr-51) release assay. Two CD19+ (Daudi, Raji) and two CD19- (K562, RPMI-8226) cell lines were labeled for 2h with Cr-51 and incubated with CART for 4h. Cr-51 release was assessed by a gamma counter. Multi-parametric flow cytometry was performed by a FACS LSR device. Results: Freezing/thawing reduced the percentage of CART generated by IL-7/IL-15 or IL-2 stimulation. Phenotypic analysis (after freezing/thawing) of CART stimulated with IL-7/IL-15 vs. IL-2 resulted in 22/12% N, 4/6% CM, 23/61% EM, 51/21% E. Lysis of CD19+ target cell lines reached a maximum of 56% (Daudi) and 40% (Raji) in the IL-7/IL-15 group. Control with non-transduced cells: 3% (Daudi), 1% (Raji). For non-transduced cells, the percentage of CD56+CD3+ NK-T cells was 15% of all CD3+ cells. Freezing of CART, stimulated with IL-2, lead to a significant decrease in the effector cell subpopulation. Thus, effector cells seem to be more vulnerable to freezing/thawing than memory cells. This decrease in the CART population might explain the decline in cytotoxicity: 38% (Daudi), 21% (Raji). Overall, CART cells seem to be more vulnerable to freezing/thawing than non-transduced cells. Resting overnight (without cytokine stimulation) did not significantly change cytotoxicity, but diminished the CART cell number with equal reduction of all subpopulations. Thus, the remaining CART became more effective during resting. Conclusions. We have established a combination of immunophenotyping and cytotoxicity assay for CART as a preclinical potency assay. Correlation of these data with the clinical outcome of patients receiving the corresponding cell products will be crucial to optimize and standardize CART therapy. Disclosure: No conflict of interest disclosed. V467

PD1-Inhibition with nivolumab after allogeneic haematopoietic cell transplantation for Hodgkin’s lymphoma

allo-HCT is unknown. We report the use of nivolumab after allo-HCT for relapsed HL. Patient and methods: The Patient is a 52-year-old female with HL, stage IVLA in 5th relapse. Earlier treatments were standard chemotherapies prior to and after autologous HCT. In 2010, the patient received an allo-HCT from an HLA-identical unrelated donor. She developed extensive graft versus host disease (GvHD) of the skin and liver. The patient suffered further relapses and was treated with brentuximab, and gemcitabine. Immunosuppression could be stopped in 2014. Peripherally, NK- and T-cells were 100% of donor origin. A PET-CT prior to nivolumab displayed a generalized lymph node, pulmonary and bone involvement by HL. Immunohistochemical staining of a tissue biopsy revealed that the Hodgkinand Reed-Sternberg cells expressed the PD-L1 protein. Results: Nivolumab was started (3mg/kg at week 1 and 4, and then every 2 weeks) in July, 2015. A PET-CT at week 5 revealed a complete remission which is now maintained for 8 months. Two weeks after the first infusion, an upsurge in eosinophils, NK- and T-cell populations occurred (table). CD279 (PD-1) was expressed on 21% of the T-cells. A grade 3 hepatotoxicity at week 6 necessitated an interruption of nivolumab for 5 weeks and treatment with prednisolone 25 mg QD. Nivolumab was resumed and prednisolone tapered without evidence of further immune-mediated adverse events or GvHD. Conclusion: A fast and complete remission with immunologic checkpoint blockade is possible even after allo-HCT. However, a therapeutic blockade of the PD-1 pathway may influence the function of lymphocytes and NKcells. Thus, further data are needed to establish the impact of PD-1/PD-L1 blockade after allo-HCT. Tab. 1. Peripheral WBC counts under nivolumab

Eosinophils (%) Lymphocytes (x10e9/L) CD3+ cells (x10e9/L) CD3+/HLA-DR+ cells (x10e9/L) CD3+/CD4+ cells (x10e9/L) CD3+/CD8+ cells (x10e9/L) CD3-/CD16/CD56+ cells (x10e9/L)

6 0.7 0.4

2 weeks after the start of nivolumab 27 1.5 1.0

5 months under nivolumab 2 1.6 1.0

0.1

0.3

0.2

0.3 0.1

0.6 0.3

0.7 0.3

0.2

0.4

0.2

Prior to nivolumab

Disclosure: No conflict of interest disclosed. V468

HLA-DPB1 specific T-cell receptors as novel approach in adoptive immunotherapy of leukemias Vatter S.1, Zwerger M.1, Mirbeth C.1, Siebörger M.1, Herr W.1,2, Thomas S.1,2 Universitätsklinikum Regensburg, Klinik und Poliklinik für Innere Medizin III, Regensburg, Germany, 2Regenburger Centrum für Interventionelle Immunologie, Regensburg, Germany 1

Allogeneic hematopoietic cell transplantation (allo-HCT) is an option for patients with Hodgkin’s lymphoma (HL) who relapse after autologous HCT. Recently, response rates of 87% after nivolumab (a monoclonal IgG4 anti-PD-1 antibody) in relapsed/refractory HL were published. Yet, the impact of nivolumab with its immune-related mechanism of action after

HLA-DPB1 mismatch antigens occur in up to 80% of allogeneic hematopoietic stem cell transplantations (HSCT) from HLA 10/10 matched donors and are associated with a significantly decreased risk of leukemia relapse. Donor T cells usually lack immune tolerance to mismatched HLA-DPB1, suggesting these antigens as promising target structures for T-cell immunotherapy in allogeneic HSCT. We have recently observed that primary acute myeloid leukemia (AML) blasts strongly expressed HLA-DPB1, and that allo-HLA-DPB1 specific AML-reactive cytolytic T lymphocytes (CTL) can be expanded in vitro from CD4+CD45RA+ naive-enriched T cells of the HLA 10/10 matched stem cell donor. The CD4 CTL lysed primary AML blasts carrying the mismatched HLA-DPB1 allele in vitro and notably eliminated them upon adoptive transfer into AML engrafted NOD/SCID/IL2Rγc-/- mice.

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–160

Jäkel N.1, Schulze S.1, Monecke A.2, Leiblein S.1, Schirmacher P.3, Niederwieser D.1, Al-Ali H.K.1 Universitätsklinik Leipzig AöR, Hämatologie/ internistische Onkologie, Leipzig, Germany, 2Universitätsklinik Leipzig AöR, Pathologie, Leipzig, Germany, 3 Universität Heidelberg, Pathologisches Institut, Heidelberg, Germany 1

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To facilitate the generation of allo-HLA-DPB1 specific CTL, we have isolated the T-cell receptor (TCR) from a highly lytic allo-HLA-DPB1*04:01 specific CD4 CTL and transferred it into T cells from healthy donors. Since this TCRDP04 recognized its antigen CD4-independently, both re-programmed CD4 and CD8 T cells effectively lysed EBV-LCL and primary AML blasts from HLA-DPB1*04:01+ patients. Detailed characterization of the TCR demonstrated its cross-reactivity to the closely related HLA-DPB1*04:02 allele, which together with HLA-DPB1*04:01 would allow for its application in about 50% of patients. Moreover, replacement of the HLA-DP α-chain revealed that recognition was specifically restricted to the HLA-DP β-chain of HLA-DPA1*01:03 /DPB1*04:01+ target cells. Further analyses revealed that TCRDP04+ T cells recognize a broad panel of human hematopoietic and non-hematopoietic as well as xenogeneic cell lines upon HLA-DPB1*04:01 transfection. From these data we hypothesize a peptide independent molecular-target structure for TCRDP04. As TCRDP04 transfected T cells recognize HLA-DPB1*04:01+ primary fibroblasts used as surrogate targets of graft versus host disease (GvHD) upon IFN-γ pre-treatment, TCRDP04 redirected T cells might induce on-target GvHD reactivity in the presence of pro-inflammatory cytokines. Introducing safety mechanisms (e.g. transient TCR expression systems) might substantially reduce the GvHD potential of TCRDP04 re-programmed T cells. Altogether, our data suggest to further investigate HLA-DPB1 specific TCRs as therapeutic ‘off-the-shelf ’ reagents in adoptive immunotherapy of leukemias.

directed against TRP2. Selected non-TRP2-reactive microcultures were further expanded and found to cross-react with up to 30% of allogeneic tumor cell lines. Conclusions: HLA I/II-negative tumor cells were successfully applied to demonstrate the presence of T cells in UCB lymphocytes that can recognize TRP2 and other target antigens in an HLA-independent manner. Therefore, UCB lymphocytes can serve as a source of HLA-independent T cells and their receptors. Their therapeutic application would allow to circumvent both immune escape mechanisms affecting HLA-restricted recognition and the dictates of HLA polymorphism.

Disclosure: No conflict of interest disclosed.

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Detection of HLA-independent tumor-reactive T cells in umbilical cord blood Pannenbeckers M.1, Derigs P.1, Fruth A.2, Paschen A.3,4, Wölfel T.1,5,6, Wölfel C.1,5 University Medical Center (UMC) of the Johannes Gutenberg-University, Internal Medicine III, Mainz, Germany, 2University Medical Center (UMC) of the Johannes Gutenberg-University, Obstetrics and Gynecology, Mainz, Germany, 3 University Hospital, University Duisburg/Essen, Dermatology, Essen, Germany, 4 German Cancer Research Consortium (DKTK), partner site Essen/Düsseldorf, Essen, Germany, 5German Cancer Research Consortium (DKTK), partner site Frankfurt/Mainz, Mainz, Germany, 6Research Center for Immunotherapy (FZI), Mainz, Germany 1

Introduction: In melanoma patient Ma-Mel-86 with clinically evident HLA I-negative metastases we have observed HLA-independent anti-tumor immune responses mediated by CD8+ αβTCR+ cytolytic T cells. Two target antigens have been identified so far. These are CSF2RA (colony stimulating factor 2 receptor alpha subunit) and TRP2 (tyrosinase-related protein 2). In order to develop assays to systematically detect, expand and analyze HLA-independent T cells also in other individuals and from different sources we generated HLA I- and II-negative cells via transcription activator-like effector nuclease (TALEN)-mediated knockout of genes B2M (encoding ß2 microglobulin) and CIITA (encoding the HLA class II transactivator). Methods: Ma-Mel-86B_KOCIITA cells (HLA I- due to naturally occurring biallelic B2M deletions and HLA II- due to TALEN-mediated CIITA knockout) were stably transfected with plasmids encoding the co-stimulatory proteins CD80 and CD83. The transfectants were used to stimulate CD3+ T cells isolated with microbeads from umbilical cord blood (UCB) lymphocytes. Responder lymphocytes were tested with IFNγ ELISPOT assays. Targets were Ma-Mel-86B_KOCIITA cells naturally expressing TRP2, CSF2RA and other yet unknown HLA-independent T cell antigens as well as HEK293T_KOB2M/CIITA cells transfected with TRP2 or CSF2RA and K562 cells. Results: UCB lymphocytes were stimulated in microcultures under quasi-limiting dilution conditions with irradiated CD80/CD83+ HLA I/IIMa-Mel-86B_KOCIITA cells. After two stimulations on day 12, 26/50 microcultures were found to recognize Ma-Mel-86B_KOCIITA but not K562. Among these we detected responders in 5 microcultures that were

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Disclosure: No conflict of interest disclosed.

Freier Vortrag

Nicht maligne Hämatologie V474

Clinical and laboratory findings in adult patients with Hemophagocytic Lymphohistiocytosis: Update from the German aHLH registry Birndt S.1, Schenk T.1, Brunkhorst F.M.2, Maschmeyer G.3, Rothmann F.3, Weber T.4, Müller M.5, Panse J.6, Schroers R.7, Braess J.8, Frickhofen N.9, Janka G.10, Lehmberg K.10, Hochhaus A.1, Ernst T.1, La Rosée P.1 Universitätsklinikum Jena, Klinik für Innere Medizin II, Hämatologie und Internistische Onkologie, Jena, Germany, 2Zentrum für Klinische Studien, Universitätsklinikum Jena, Jena, Germany, 3Klinikum Ernst von Bergmann, Klinik für Hämatologie, Onkologie und Palliativmedizin, Potsdam, Germany, 4 Klinik für Innere Medizin IV, Hämatologie und Onkologie, Universitätsklinikum, Halle (Saale), Germany, 5Zentrum für Infektiologie und HIV, Vivantes AugusteViktoria-Klinikum, Berlin, Germany, 6Universitätsklinikum RWTH Aachen, Klinik für Hämatologie, Onkologie, Hämostaseologie und SZT, Aachen, Germany, 7 Medizinische Universitätsklinik Knappschaftkrankenhaus, Bochum, Germany, 8 Krankenhaus Barmherzige Brüder Regensburg, Onkologie und Hämatologie, Regensburg, Germany, 9HELIOS Dr. Horst Schmidt Kliniken Wiesbaden, Innere Medizin III: Hämatologie, Onkologie und Palliativmedizin, Wiesbaden, Germany, 10 Nationales Studienzentrum pädiatrische HLH, Universitätsklinikum HamburgEppendorf, Hamburg, Germany

Introduction: HLH is a severe hyperinflammatory syndrome. In contrast to HLH in children, HLH in adults (aHLH) presents with a wider spectrum of trigger diseases (infections, malignancies and autoimmune diseases). Genetic cofactors (e.g. heterozygous mutations of immune regulating genes like perforin (PRF1)) only recently came into focus in aHLH. As systematic data on aHLH are not available in Germany, a national multicenter registry was initiated (www.hlh-registry.org) in August 2010. Targeted sequencing of PRF1 was performed depending on sample availability. Methods: Patients (pts) with suspected aHLH were entered by 29 institutions. HLH was confirmed using the HLH-2004 criteria. Clinical and laboratory findings as well as treatments and outcomes were recorded. The H-Score (www.saintantoine.aphp.fr/score/) was used to validate HLH diagnosis. PRF1 sequencing was performed in 21/107 pts. Results: 107 pts (41 female) with a median age of 48 years (range 17–81) were registered from August 2010 to April 2016. 83/107 (78%) fulfilled the HLH-2004 criteria. Trigger diseases were neoplasia in 28/83 pts (lymphoma n = 24, acute leukemia n = 3, other malignoma n = 1), infections in 29/83 pts with EBV being the most common, and autoimmune/-inflammatory diseases in 10/83 pts. 5/83 pts developed HLH after allogeneic stem cell transplantation (alloSCT), of whom 4/5 pts showed EBV infections and one patient a CMV infection. A trigger could not be identified in 11/83 pts. 4/107 pts presented with HLH-like symptoms triggered by systemic leishmaniasis. After a median follow up of 110 days (range 1–2300), 47/83 pts (57%) were alive. Treatment included corticosteroids in 60/83 pts, etoposide in 39/83 pts and i.v. immunoglobulins in 33/83 pts. Genotyping revealed late onset hereditary HLH in 3 pts (1 pt CMV/ EBV-HLH, XLP-2; 1 pt EBV/H1N1-HLH, XLP-1; 1 pt unknown trigger, compound heterozygous PRF1 mutation). A heterozygous PRF1 mutation

Abstracts

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(A91V) was found in 2 pts with aHLH (1 pt after alloSCT, 1 pt with lymphoma associated aHLH). 4/5 pts with HLH-associated mutations died within three months. Conclusions: Our data confirm the heterogeneity of trigger diseases of aHLH in Germany. Vigilance towards leishmaniasis in patients presenting with HLH-like symptoms is strongly recommended to protect these patients from harmful immunosuppression. In addition to known trigger diseases, inherited predisposition may contribute to HLH susceptibility and prognosis in adults. Disclosure: No conflict of interest disclosed. V475

The terminal complement inhibitor eculizumab in Cold Agglutinin Disease (CAD): A prospective phase II trial (DECADE Trial) Röth A.1, Bommer M.2, Hüttmann A.1, Herich-Terhürne D.1, Kuklik N.3, Lenz V.4, Schrezenmeier H.5, Dührsen U.1 Universitätsklinikum Essen, Klinik für Hämatologie, Essen, Germany, Universitätsklinikum Ulm, Klinik für Innere Medizin III, Ulm, Germany, 3 Universitätsklinikum Essen, Institut für medizinische Informatik, Biometrie und Epidemiologie, Essen, Germany, 4Universitätsklinikum Essen, Institut für Transfusionsmedizin, Essen, Germany, 5Universitätsklinikum Ulm, Institut für Klinische Transfusionsmedizin und Immungenetik Ulm, Ulm, Germany 1 2

Background: Cold agglutinin disease (CAD) is a rare autoimmune hemolytic anemia (AIHA). Many conventional therapeutic approaches used in other AIHA or indolent lymphomas are largely ineffective and the therapeutic efficacy of rituximab is limited. Successful long term treatment of a patient with CAD with eculizumab (Röth et al., Blood 2009) suggested testing this novel approach in a larger cohort. Patients and methods: A prospective, controlled, non-randomized, multi-center, phase II trial with pre- and post-eculizumab treatment comparison was conducted (NCT01303952). Main inclusion criterion was symptomatic hemolysis. The primary endpoint was the LDH level, secondary endpoints included transfusion requirements, parameters of intravascular hemolysis, quality of life, 6-minute walk test (6MWT), circulatory symptoms and thromboembolic events (TE). The analysis was performed in the intention-to-treat population. Results: Between 2011 and 2014 13 treatment-naïve or pretreated patients with CAD were enrolled. The median age was 74 years (range: 64 - 80). After treatment with eculizumab the LDH decreased from a median pre-treatment level of 572 U/L (mean ± standard deviation: 670 ± 300 U/L) by an average of 56% to a median end-of-treatment level of 327 U/L (400 ± 279 U/L; p = 0.0215, Wilcoxon signed-rank test). There was an improvement of anemia and a reduction in transfusion requirement from a median of 4.0 red blood cell units per patient in the 12-month period preceding participation in the study to 0.0 units during treatment with eculizumab. The result of the 6MWT improved from a median of 358 m at baseline to a median of 420 m at the end of treatment (p = 0.078). During the enrollment phase, 7 TEs including 1 pulmonary embolism were recorded in a total of 4 patients; no TEs were observed during eculizumab treatment. Conclusions: Treatment with eculizumab was safe and well tolerated. A statistically significant reduction in hemolysis and transfusion requirements was observed with a favorable impact on thromboembolic complications. Terminal complement activation seems a relevant mediator of erythrocyte destruction in CAD. Lacking effective treatment options eculizumab is the treatment of choice for severe hemolysis in CAD.

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‘Gardos Channelopathy’ - a variant of hereditary Stomatocytosis with complex molecular regulation Kaestner L.1,2, Fermo E.3, Bogdanova A.4, Petkova-Kirova P.1, Zaninoni A.3, Marcello A.P.3, Makhro A.4, Hänggi P.4, Hertz L.1, Danielczok J.1, Vercellati C.3, Mirra N.5, Zanella A.3, Cortelezzi A.3,6, Barcellini W.3, Bianchi P.3 Universität des Saarlandes, Forschungsstelle für Molekulares Imaging und Screening, Homburg/Saar, Germany, 2Experimental Physics, Living Fluids, Saarbrücken, Germany, 3UOC Oncoematologia, UOS, Fisiopatologia delle Anemie Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milano, Italy, 4University of Zürich, Institute of Veterinary Physiology, Vetsuisse Faculty and the Zürich Center for Integrative Human Physiology (ZIHP), Zürich, Switzerland, 5UOC. Pediatria a media intensità di cura, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milano, Italy, 6Universita’ degli Studi di Milano, Milano, Italy 1

Introduction: The Gardos channel is a Ca2+ sensitive, K+ selective channel present in several tissues including red blood cells, where it is involved in cell volume regulation. Activation of the channel in response to elevation of cytosolic Ca2+ in human red blood cells causes transient cell shrinkage due to the efflux of K+. Recently, mutations at two different amino acidic residues in KCNN4 have been reported in patients with hereditary Xerocytosis. Methode and Results: By whole exome sequencing we identified a new family with two patients affected by chronic hemolytic anemia carrying mutation R352H in the KCNN4 gene. To evaluate the effects of this mutation on the cellular properties and eventually on the clinical phenotype, we performed functional studies in erythroid precursors and mature RBCs of the two patients. Gardos channel hyperactivation was demonstrated not only in circulating RBCs, but also tracked down to erythroblasts differentiated ex vivo from peripheral CD34+ cells. Pathological alterations in function of multiple ion transport systems including Na,K-ATPase and Cl-dependent K+ cotransporters were also observed, resulting in compensatory effects ultimately preventing cellular dehydration, in line with the observation of normal MCHC and normal Osmoscan curve in the two patients. Moreover, by flow cytometry and confocal fluorescence live-cell imaging we demonstrated Ca2+ overload in RBCs of both patients and hypersensitivity of Ca2+ uptake by RBCs to swelling, resulting in a further increased activity of mutated Gardos channel. Conclusions: These findings suggest that the ‘Gardos channelopathy’ is a complex pathology and to some extent different from the common hereditary Xerocytosis. Disclosure: No conflict of interest disclosed. V477

Eculizumab rescue therapy in AB0-incompatible red blood cell transfusion: a case report Röth A.1, Dirkmann D.2, Lenz V.3, Heinrichs S.3, Fuchs N.3, Breyer M.3, Dührsen U.1, Peters J.2, Horn P.A.3 Universitätsklinikum Essen, Klinik für Hämatologie, Essen, Germany, Universitätsklinikum Essen, Klinik für Anästhesiologie und Intensivmedizin, Essen, Germany, 3Universitätsklinikum Essen, Institut für Transfusionsmedizin, Essen, Germany 1 2

Disclosure: Alexander Röth: Advisory Role: Alexion, Novartis, True North; Financing of Scientific Research: Alexion; Expert Testimony: Alexion Ulrich Dührsen: Financing of Scientific Research: Alexion; Expert Testimony: Alexion

Background: With 4–5 million units of blood annually transfused in Germany, the estimated incidence of erroneously administered transfusions is 1:12,000–1:38,000. Activation of the complement system by isoagglutinins (anti-A, anti-B; mainly immunoglobulin M) after transfusion of AB0-incompatible red blood cells (RBCs) can cause severe and lethal complications. The anti-C5 monoclonal antibody eculizumab can block activation of the complement cascade and prevent hemolysis of incompatible RBCs, otherwise evoking release of free hemoglobin and anaphylatoxins with disastrous consequences. A beneficial effect of eculizumab in this clinical scenario was reported previously (Weinstock et al., 2015). Patient and Results: A 21year old male patient with blood group 0 was mistakenly transfused a whole unit of A1 RBCs during surgery. At the

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end of the transfusion the patient developed hemoglobinuria and after extubation also chills and malaise. Within 2½ hours he was treated with 900 mg eculizumab intravenously, forced diuresis, and urine alkalinization. Preexisting antibiotic treatment was continued to cover Neisseria meningitides for 4 weeks. Otherwise, the patient remained in stable cardiopulmonary conditions. Initial hemoglobinuria disappeared immediately. LDH activity increased from a normal baseline preoperatively to a maximum of 809 U/l on the day of transfusion (d0) and had normalized on d4. Haptoglobin was initially depleted (< 0,01 g/l) but also normalized by d4. Hemoglobin concentration decreased from 8.8 g/dl to a minimum of 7.1 g/dl on d1. A maximum total bilirubin concentration of 5.7 mg/dl was reached on d0. As shown by flow cytometry, type A RBCs reached a maximum relative concentration of 10.8% on d0 and had vanished in the peripheral blood by d4. The monospecific DAT became positive for IgG, C3c, and C3d until d2. Summary and Conclusions: This is the first report of a non-PNH patient treated with eculizumab for an AB0-incompatible packed red cell transfusion. Immediate treatment with eculizumab apparently decreased intravascular hemolysis and its grave consequences. Type A RBCs were protected from a terminal complement attack and thus remained detectable for several days within the circulation. Due the risk of severe, life-threatening complications by incompatible transfusions, the lack of other options, and relatively minor side effects of eculizumab such a rescue therapy appears a rational treatment if an acute maltranfusion is recognized. Disclosure: Alexander Röth: Advisory Role: Alexion; Financing of Scientific Research: Alexion; Expert Testimony: Alexion Peter Horn: No conflict of interest disclosed. V478

Evaluation of non-invasive MRI assessment after Gondon & Rose for hepatic iron stores – results of the first 60 assessments Kragl B.1, Henze L.1, Große-Thie C.1, Kriesen U.1, Sell K.2, Hauenstein K.2, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany, 2Institution for Diagnostic and Interventional Radiology, Rostock University Medical Center, Rostock, Germany 1

Introduction: Patients receiving multiple blood transfusions frequently develop iron overload due to the fact that the human body is not able to eliminate large quantities of iron. Excessive iron accumulates in the liver, spleen, heart, and endocrine organs. Diagnostic serum ferritin level normally correlate to body iron, but might be altered incorrectly in patients with malignancies, inflammations or hepatic lesions. Herein we evaluated a non-invasive MRI based approach to assess liver iron stores. Methods: A non-invasive MRI assessment of the liver after Gondon & Rose was performed in 58 patients from November 2011 to January 2016. MRI was analysed in regards to patient characteristic (sex, age, underlying disease) and patient serum ferritin level. Results: In total 60 MRI assessments from 58 patients were conducted. Data of 57 assessments were included. Median age of the patients was 56 years (13–71 years). Most patients were male (58.6%) vs. female (41.4%). Following diseases entities were represented: acute leukemia and myelodysplastic syndrome (51.7%), lymphoma (17.2%), anemia (8.6%), myeloproliferative neoplasms (6.9%), myeloma (5.2%), solid tumors (5.2%), hemochromatosis (5.2%). Most patients had received >15 RBC units. Iron load in all assessments ranged from 22 µmol/g up to 372 µmol/g (normal range < 150 µmol/g). Only 14.0% of the assessments showed no iron overload (< 150 µmol/g). A moderate iron overload (150–265 µmol/g) was detected in 43.8%, while 22.8% showed a intermediate overload (266–300 µmol/g). A severe iron overload (>300 µmol/g) was detected in 19.3% of the assessments.

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Median ferritin level was 3016 mg/l (43–55048 mg/l). According to ULN graduation (2.5x/5x/10x/>10x) grouping was as follows 20%, 17%, 38% and 26% respectively. 50% of patients with elevated levels of ferritin > 2.5 ULN showed no iron overload, while 90% of patients with severe MRI iron load showed levels > 2.5 ULN. Conclusions: Evaluation of the analysed cohort revealed that non-invasive assessment of hepatic iron stores by MRI can be used as a diagnostic tool. This method represents a valuable addition to objectify a suspected organ siderosis especially in patients with high ferritin levels due to other reasons. Accordingly MRI based screening can be used to confirm a diagnosis as well as to monitor a manifest hepatic iron overload. Disclosure: No conflict of interest disclosed. V479

Frustrating results from long term treatment with TPOs in patients with ITP: A balance of 10 years observation Depré F.1, Salama A.1 Charité Berlin, Institute of Transfusionsmedizin, Berlin, Germany

Background: Based on the results of well designed studies, it is generally believed that both thrombopoietin receptor agonists (TPOs) romiplostim and eltrombopag are highly effective and safe for treatment of ITP. In practice, these drugs appear to be ineffective and/or intolerable more frequently than has yet been suggested. Patients and methods: During the last decade a total 89 patients were treated with eltrombopag and/or romiplostim from one hand at our institution. Results: Initially, the majority of treated patients (≥70%) were found to be responders to TPOs. However, in addition to non-responding patients, many of the responders developed intolerable adverse reactions including arthralgia/myalgia (n = 10), gastrointestinal reactions (n = 5), vision impairment (n = 3), vasculitis (n = 2) and many other different reactions, that led to discontinuation of the drugs despite response. Cross reactive adverse reactions and/or new adverse reactions were observed in 8 (24%) of 34 patients following a switch to the alternate TPO. Ultimately, some of the patients who continued treatment with TPOs (eltrombopag 16 of 73 patients and romiplostim 19 of 49 patients) may also develop intolerable adverse reactions during further observation. Conclusion: As both TPOs not only do not cure patients with ITP, but may also cause severe side effects, a strict and continuous surveillance throughout the course of treatment is invariably necessary. Most importantly, the need for alternative treatment options remains obvious. Disclosure: No conflict of interest disclosed.

Freier Vortrag

(Minimale) Resterkrankung nach Leukämietherapie V480

Comparative quantification of BCR-ABL Transcripts and IG/ TR gene rearrangements during the course of the therapy in Philadelphia chromosome-positive adult ALL (PH+ALL) Meißner A.-M.1, Brüggemann M.1, Trautmann H.1, Gökbuget N.2, Kneba M.1, Pfeifer H.2 University Hospital Kiel, Department of Hematology and Oncology, Kiel, Germany, 2Goethe University Hospital, Department of Medicine II, Hematology/ Oncology, Frankfurt/M., Germany 1

Introduction: Quantification of minimal residual disease (MRD) is a well-established tool for risk stratification in acute lymphoblastic leukemia (ALL). While the molecular MRD quantification for PH negative adult ALL is performed by the allele-specific real-time quantitative (ASO RQ-) PCR of immunoglobulin and T-cell receptor (IG/TR) gene rear-

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rangements, the quantification for PH+ALL usually relies on the detection of BCR-ABL transcripts. Until now only few comparisons between those two methods exist. Those however imply certain differences. Therefore we performed a systematic comparison of both methods applying standardized quantification methods. Methods: We analyzed and compared MRD levels quantified by IG/TR based MRD assessment and BCR-ABL transcript detection in left-over samples of 106 adult patients with PH+ALL being treated within the German Acute Lymphoblastic Leukemia Multicenter Study Group (GMALL) protocols. MRD measurements were carried out by two reference institutions for MRD diagnostics in Germany according to EuroMRD guidelines. Correlation was determined by the Spearman Rank Correlation Coefficient. Results: A total of 147 samples was analyzed (27 blood and 120 bone marrow samples) by both methods in parallel. A total of 40/147 samples (27.2%) showed double negative results, 58 samples (39.5%) were double positive, including 9 samples which were positive below the quantitative level (pos< Q) of IG/TR RQ-PCR. While 12 samples (8.2%) were positive by IG/TR MRD (thereof 6 pos< Q) and negative by BCR-ABL MRD, 37 (25.2%) samples were positive by BCR-ABL and negative by IG/TR. The overall correlation was only moderate with r2= 0.6221 (p-value < 0,0001) with best correlation at early treatment time-points. Generally, IG/TR ASO RQ-PCR converted to MRD negativity earlier compared to BCRABL RQ-PCR. Notably, 24 of the IG/TR negative samples showed a fairly high amount of BCR-ABL ranging from 1E-04 to 1E-01 although sensitivity of IG/TR RQ-PCR reached 1E-04 to 1E-05. Also 7 samples which were pos< Q for IG/TR showed BCR-ABL values of 1E-04 to 1E-02. Conclusion: BCR-ABL and IG/TR-based MRD monitoring in PH+ALL show only moderate correlation. A potential explanation for the discrepancy of IG/TR negative samples showing fairly high BCR-ABL MRD values is a multilineage involvement of BCR-ABL whereas IG/TR gene rearrangements are lineage restricted. The clinical relevance of this finding has to be clarified by correlation to outcome.

chemotherapy cycles) showed no differences between MRD+ & MRDpts, except for a trend for lower peripheral blast countdx in MRD+ group (P = 0.07). 11 pts suffered relapse after HCT of which 9 (81.8%) were MRD+. 4 patients did not experience relapse but were MRD+, 2 of them died within 100 days after HCT due to treatment-related complications. Pre-HCT MRD+ was associated with a significantly higher cumulative incidence of relapse (P < 0.001, Figure1A) & shorter overall survival (P = 0.004, Figure1B). NPM1mutA as a MRD marker pre-HCT is a strong prognosticator independent of the clinical, cytogenetical & molecular context. AML pts with MRD+ have significant worse outcome although transplanted in CR. NPM1mut monitoring before HCT should guide therapy & prospective trials will maybe show that MRD+ pts might benefit from an additional cycle chemotherapy before HCT.

Disclosure: No conflict of interest disclosed.

Persistence of non-DNMT3A mutations predicts relapse in cytogenetically normal AML

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Gaksch L.1, Kashofer K.2, Heitzer E.3, Daga S.1, Hofer S.1, Krisper N.1,4, Höfler G.2,4, Zebisch A.1, Sill H.1, Wölfler A.1,4

NPM1 Type A mutations as minimal residual disease marker in acute myeloid leukemia patients before allogeneic stem cell transplantation determined by highly sensitive and specific digital droplet PCR is a strong prognosticator for outcome Bill M.1, Kloss L.1, Jentzsch M.1, Grimm J.1, Schubert K.1, Schulz J.1, Knyrim M.1, Cross M.1, Vucinic V.1, Behre G.1, Pönisch W.1, Franke G.-N.1, Niederwieser D.1, Schwind S.1

Fig. 1. Disclosure: No conflict of interest disclosed. V482

Medical University of Graz, Division of Hematology, Graz, Austria, 2Medical University of Graz, Institute of Pathology, Graz, Austria, 3Medical University of Graz, Institute of Human Genetics, Graz, Austria, 4CBMed, Center for Biomarker Research in Medicine, Graz, Austria 1

Relapse is a major mortality cause in acute myeloid leukemia (AML) patients (pts) undergoing allogeneic hematopoietic stem cell transplantation (HCT). Identifying pts with high relapse risk is crucial for treatment guidance. Minimal residual disease (MRD) assessment, based e.g. on NPM1 mutation (mut) detection, has been shown to be of clinical relevance but data in HCT pts are limited. We identified 46 pts (median age 64.5 years (y), range 33.1–74.1y) with NPM1mutA that received HCT after myeloablative (Cyclophosphamide + 12Gy total body irradiation [TBI], n = 11) or non-myeloablative (Fludarabine + 2Gy TBI, n = 35) conditioning at our institution between 2000 & 2015. All had available bone marrow (BM) samples pre-HCT (within 30 d before HCT) & were in complete remission with (CR; n = 42; 91.3%) or without complete hematological recovery (CRi; n = 4; 8.7%). NPM1mutA copy numbers were measured on cDNA level by ddPCR & normalized to ABL1. Those with a ratio >0.01% or ≥3 positive (+) droplets were defined as + according to manufacturer’s recommendations. Pre-HCT, we found 13 (28.3%) out of 46 pts to be MRD+ (NPM1mutA/ABL1 range 0.014%-612.88%). Analyzing several clinical & molecular characteristics (i.e. agedx, sex, ELN distribution, Hbdx, Plateletsdx, WBCdx, BM blastsdx, CR1 vs CR2 vs CRi, CEBPAmut, FLT3-ITD, number of

Introduction: Despite achieving complete remission after intensive therapy most patients with cytogenetically normal AML relapse due to the persistence of submicroscopical residual disease. We asked whether parallel sequencing of DNA isolated from bone marrow slides displaying complete remission allows the detection of persisting AML-associated mutations identifying patients at increased risk for relapse. Methods: We included 34 AML patients of whom diagnostic material and slides after at least one cycle of consolidation were available. Isolated DNA was screened for mutations in 19 genes using an Ion Torrent sequencing platform. Persistence of a mutation was defined by a z-score >3 for the mutated allele using the error rate of non-mutated samples as a reference. In addition, the variant allelic frequency of distinct mutations was validated by digital PCR and parallel sequencing using a barcoding approach. Results: Three patients either revealed no mutation in the diagnostic material or the somatic origin of the identified mutations could not be proven. In addition, DNA isolated from four slides was not sufficient for sequencing. Therefore, in total 27 patients could be analyzed for mutation clearance. In these patients we identified 68 somatic mutations (median of 3 mutations per patient, range 1–5) and 22 of these were still detected in 16 patients after consolidation therapy (median one mutation, range 0–3). The determined variant allelic frequencies of NPM1 or DNMT3A correlated well when compared with either digital PCR or a barcoding parallel sequencing approach, respectively. The most frequent non-cleared mutations were found in DNMT3A (n = 10 pts). Since these are known to persist in preleukemic stem cells

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Universitätsklinikum Leipzig, Abteilung für Hämatologie und Onkologie, Leipzig, Germany 1

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without any impact on relapse risk, we performed survival and relapse risk analysis excluding DNMT3A mutations. Importantly, persistence of non-DNMT3A mutations was significantly associated with a higher risk of AML relapse (7 out of 8 pts versus 6/19 pts; p = 0,013) and with a shorter relapse-free survival (282 days vs. not reached; log-rank p = 0,024). In addition, there was a trend for worse overall survival (log-rank p = 0,050). Conclusions: Persistence of non-DNMT3A mutations after consolidation therapy as detected by parallel sequencing is associated with an increased risk of relapse in cytogenetically normal AML. Disclosure: No conflict of interest disclosed. V483

DNMT3A mutation in AML patients after allogeneic stem cell transplantation Behrenbeck F.1, Blau I.W.1, Berenstein R.1, Fransecky L.1, Baldus C.1, Arnold R.1, Pezzutto A.1, Doerken B.1, Blau O.1 Charité University Berlin, Department of Hematology, Oncology and Tumorimmunology, Berlin, Germany 1

Introduction: DNMT3A mutation is one of the most frequent somatic mutations in acute myeloid leukemia (AML). Although DNMT3A mutations have been studied extensively, their role on the pathogenesis of AML remains not completely clear. Little is known about the significance of mutations burden in the time of diagnosis on the clinical and prognostic features of AML. Recently discovered persistence of DNMT3A mutations in complete remission (CR) after standard therapy indicates the presence of mutation in early pre-leukemic stem cells. In this aspect, it would be essential to analyze the existence of DNMT3A-positive cells after after allogeneic stem cell transplantation (alloSCT). Methods: Using quantitative PCR, we analyzed the burden of DNMT3A mutations in diagnostic samples obtained from 55 AML patients. Then follow up samples were analyzed repeatedly, after induction, consolidation therapy, and after alloSCT with a mean follow-up of 23 months. Using well-established markers of molecular remission and donor engraftment, we monitored DNMT3A stability during CR. Results: We have found considerable variation of mutations burden (from 1% till 90%) in diagnostic samples. Although we have not verify prognostic impact of DNMT3A mutations burden in diagnostic samples, we have found significant associations higher mutations burden with M4/M5 FAB variant of AML (p = 0,04) and presence of NPM1 mutation (p = 0,023). Persistence of DNMT3A mutations in CR after standard therapy was found in all patients. However, in CR after alloSCT in patients with complete donor chimerism, we have no discovered DNMT3A mutation. This data suggests the removal of leukemic stem cells after alloSCT and indicate the importance of alloSCT for high risk AML patients. In relapse of leukemia, all samples showed an increasing of DNMT3A mutated alleles. The loss of correlation between DNMT3A and others mutations in CR after standard chemotherapy could be explained by the presence of mutations in different leukemic subclones. Conclusion: We conclude that quantitative detection of DNMT3A mutations at different time points of AML disease could provide additional prognostic information and could explain the role of mutations in development and progression of AML. Disclosure: No conflict of interest disclosed.

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Longitudinal monitoring of Wilms’ Tumor 1 (WT1) expression by a standardized European Leukemia Net (ELN)-certified assay for detection of minimal residual disease (MRD) in MDS patients undergoing allogeneic blood stem cell transplantation Rautenberg C.1, Pechtel S.1, Hildebrandt B.2, Betz B.2, Dienst A.1, Nachtkamp K.1, Schmidt P.V.1, Heyn C.1, Kondakci M.1, Geyh S.1, Haas R.1, Germing U.1, Kobbe G.1, Schroeder T.1 Universitätsklinikum Düsseldorf, Klinik für Hämatologie, Onkologie und Klin. Immunologie, Düsseldorf, Germany, 2Universitätsklinikum Düsseldorf, Institut für Humangenetik, Düsseldorf, Germany 1

Introduction: Measurement of WT1 mRNA expression can be employed for quantitative MRD monitoring in >90% of de-novo AML pts undergoing allo-SCT. WT1 overexpression has also been reported in a relevant proportion of MDS pts, suggesting it’s potential use as MRD marker also in MDS. Still, this has not completely been established due to limited pts numbers, differing sample sources and non-standardized methods. This prompted us to investigate PB WT1 expression as MRD marker in MDS and to correlate the results with other methods commonly employed for post-transplant monitoring. Methods: A total of 37 pts with MDS (n = 24) and sAML (n = 13) who displayed WT1 overexpression at diagnosis and underwent allo-SCT were included in this retrospective study. WT1 levels were measured in PB using the ELN-certified Ipsogen® WT1 ProfileQuant® Kit. STR-based chimerism and XY-FISH (n = 19) was routinely monitored after transplant. In addition, in pts with a disease-specific cytogenetic marker (n = 25) conventional cytogenetics and FISH were performed, while in 6 pts with molecular aberrations NGS- or qPCR-based monitoring was used. Results: Median follow up of all pts was 13 months (5 to 40). During post-transplant period we analyzed a total of 296 WT1 samples corresponding to a median of 7 samples per patient (2 to 19). All 37 pts achieved complete hematological remission one month after transplant accompanied by a rapid decline of WT1 levels below normal threshold. Nineteen patients experienced hematological (63%) or molecular relapse (37%) after a median of 168 d (28 to 937) following allo-SCT. Of these, 16 showed at least one WT1 level above the cut-off level at/or before relapse (sensitivity 84%), while 3 pts with molecular relapse had persistently low WT1 levels. In pts with hematological relapse and no preemptive intervention median time from first positive WT1 value and relapse was 3 weeks (0 to 27). Of 18 pts (49%) who remained in CR for a median of 15 months (4 to 40), 17 pts displayed persistent negativity for WT1, whereas one patient became transiently positive (specificity 95%). This stable relation between WT1 expression levels and post-transplant disease burden was also reflected by a clear correlation with the results from chimerism, cytogenetic and NGS analysis. Conclusion: These findings suggest that PB WT1 expression can be used as MRD marker in MDS pts for relapse detection and guidance of therapeutic interventions after allo-SCT. Disclosure: No conflict of interest disclosed. V485

Monitoring of Calreticulin mutations and hematopoietic chimerism by digital PCR after allogeneic stem cell transplantation in patients with myelofibrosis Badbaran A.1, Fehse B.1, Abd Kadi S.S.S.1, Christopeit M.1, Aranyossy T.1, Ayuk F.1, Wolschke C.1, Kröger N.1 Universitätsklinikum Hamburg Eppendorf, Stammzelltransplantation, Hamburg, Germany 1

Myelofibrosis (MF) is a myeloproliferative disorder often associated with poor prognosis. Allogeneic stem cell transplantation (aSCT) remains the only potentially curative treatment for MF. Mutations in JAK2, MPL, and

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Calreticulin (CALR) have been described as initiating events. Molecular monitoring for MRD after aSCT has been suggested as a vital tool to guide possible adoptive immunotherapy. CALR mutations occur in 30% of all MF patients and 80% of JAK2- and MPL-negative patients. Two types were identified: type 1 represents a 52bp deletion producing the protein change p.L367fs*46 and type 2 results from a 5-bp insertion (p.K385fs*47). The prognosis of CALR mutated PMF does not depend on the type of mutation (Christopeit et al., 2016). To monitor MRD we here used digital-PCR techniques described for CALR type-1 (Mansier et al.,J Mol diag. 2016) and type-2 (Badbaran et al., BMT 2016) mutations. MRD data was compared with chimerism data, also obtained by digital PCR. Out of 143 patients with MF, 92 were JAK2V617F positive, 4 MPL positive and 35 CALR positive (21 harbored type-1 and 8 the type-2 mutation). Samples for follow-up analyses were available for eight patients with type1 and seven with type-2 mutation. Dynamics of CALR mutation levels after aSCT were in excellent agreement with clinical disease status. For CALR type 1, three patients reached MRD negativity within two months and remained in stable complete molecular remission (CMR) since. In the other five patients, dPCR revealed increasing levels of CALRt1-positive cells indicative of MRD. In four of them, ceasing immune suppression was sufficient to establish CMR and full chimerism. In contrast, the fifth patient required therapeutic intervention (HSC boost) due to a high increase (>30%) in CALRt1-positive cells, but went into CMR thereafter. Similar data was found for CALR type 2. Three patients became MRD-negative after 3 and two after 6 months. All remained in CMR since. The two other patients required intervention due to increasing numbers of CALRt2-positive cells in conjunction with decreasing chimerism. While one patient achieved CMR after ceasing immune suppression, the other eventually relapsed, but was successfully treated with a second aSCT and is now in CMR. In conclusion, both type-1 and type-2 CALR mutations represent sensitive MRD markers for follow-up diagnostics after aSCT. Digital PCR is an excellent method for precise monitoring of CALR mutations and chimerism. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium

Wirkungen und Nebenwirkungen im Versorgungsalltag V488

Quality of Life and other outcomes in the framework of complex interventions Klinkhammer-Schalke M.1, Lindberg P.1, Koller M.2, Steinger B.1 Universität Regensburg, Institut für Versorgungsforschung und Qualitätssicherung, Regensburg, Germany, 2Zentrum für Klinische Studien, Regensburg, Germany 1

Introduction: Medical treatment in patient-centred care in oncology is broadly managed and regulated in terms of guideline development, cancer centres and control of quality of outcomes by cancer registries. In contrast to this quality management cycle, there are no equal standards for patient-reported outcomes like quality of life (QoL). Therefore the Tumor Center Regensburg, Institute for Qualitymanagement and Health Care Research,designed and implemented in a complex intervention a QoL pathway with diagnosis and therapy (multidimensional therapeutic network), demonstrating effectiveness for patients with breast cancer. Methods: To provide local tailored QoL-diagnosis and -therapy to other cancer patients as well, external validity is extended by adapting the QoL-pathway to another cancer type. QoL-pathway will be adapted for colorectal cancer patients (CRC), testing its effectiveness in a pragmatic randomized controlled trial. 220 primary

Abstracts

CRC patients, surgically treated in one of four participating hospitals, will be included. QoL is measured in all patients 0, 3, 6, 12 and 18 months after surgery (EORTC QLQ-C30, QLQ-CR29). In intervention group, QoL-scores are transformed into a QoL-profile. Control group receives follow-up according to S3-guideline without profile or expert report. At primary endpoint (12 months) rate of patients with diseased QoL is compared in both groups. Results: In a complex intervention a clinical pathway with QoL diagnosis and QoL therapy has been developed, implemented and the effectiveness has already been demonstrated for breast cancer patients: At the prospectively defined and published 6 months end point 60 (95% C.I. 51 to 68) of 85 patients in the control group or 71% showed diseased in QoL in at least one dimension. In the treatment group this occurred in 47 (95% C.I. 38 to 56) of 84 patients or 56% (Χ²-test p = 0.048). This corresponded to a 21% relative risk reduction (95% C.I. 0 to 37), 15% absolute risk reduction (95% C.I. 0.3 to 29) and an NNT of 7 (95% C.I. 3 to 37) . Conclusion: A three-component outcome system including traditionally objective and patient reported subjective endpoints with qualitative analysis of clinical relevance was developed over the last 10 years.It was evaluated stepwise according to the MRC framework Disclosure: No conflict of interest disclosed.

Debatten

Ist die Zeit reif für eine therapiefreie Remission bei der CML? V492

Is the time ripe for a treatment-free remission in CML ? Pro : discontinuation of TKI in clinical routine von Bubnoff N.1 Universitätsklinik Freiburg, Med. Klinik I, Freiburg, Germany

Life-long TKI treatment in CML leads to a diminished quality of life, toxicities and considerable cost. Data from several studies, including the French STIM Study and the European EURO-SKI study show that in patients with ongoing molecular response, a treatment-free remission can be achieved in about half the cases. These studies have also shown that treatment interruption is safe: Frequent (every 4 weeks) PCR monitoring allows early detection of molecular relapse (PCR 0.1% or above) and immediate re-institution of treatment. Virtually all patients achieved previous responses after re-institution of TKI treatment. An increased risk of disease progression after TKI discontinuation at the present time is not supported by available data, and thus is a theoretical risk. Highly sensitive BCR-ABL PCR is now widely available, so TKI interruption and monitoring of treatment-free remission is feasible in routine practice. Thus, available data support discontinuation of TKI in the clinical routine setting. To meet this requirement, specific recommendations for treatment discontinuation in CML outside of clinical trials should be provided. Disclosure: Nikolas von Bubnnoff: Financing of Scientific Research: Fa. Novartis, Fa. BMS; Expert Testimony: Fa. Novartis V493

Contra: TKI discontinuation as routine clinical pratice Burchert A.1, CML-Studiengruppe Universitätsklinikum Marburg, Klinik für Hämatologie, Onkologie und Immunologie, Marburg, Germany 1

There is accumulating evidence to suggest that Abl-kinase inhibitor (TKI) discontinuation is safe in a selected cohort of CML patients. The largest TKI discontinuation trial, the EURO-SKI study, included more than 600 patients in Europe and confirmed that approximately 50% of the patients in deep molecular remission (MR4 superscript or better) will eventually maintain a treatment free remission. However, this author argues contra

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the motion to discontinue TKI in routine clinical practice outside of a clinical trial. There is lack of knowledge about the molecular mechanisms of disease recurrence after TKI stop and lack of long-term safety data for CML patients without treatment. There must be awareness that disease recurrence and progression remains a realistic risk – particularly in patients, who remain BCR-ABL mRNA-positive after TKI stop. There is currently no consensus regarding who is an optimal TKI-STOP candidate and how such a patient should be monitored after TKI cessation. However, making these decisions cannot be left to the discretion of individual hematologists in a hospital or private practice. With a life-saving and very safe treatment standard (TKI) at hand, it appears therefore ethically debatable, whether at this time available data are mature enough to declare TKI discontinuation in whatever remission at whatever time point as a new and safe CML treatment standard. Disclosure: Andreas Burchert: Financing of Scientific Research: Bristol-Myers-Squibb, Pfizer

Posterdiskussion

Akute myeloische Leukämie, experimentell P494

A focused CRSPR/CAS9 screen identifies snoRNAs that are required for clonal growth of leukemia cells Pauli C.1, Liu Y.1, Zhou F.1, Gerloff D.1, Rohde C.1, Müller-Tidow C.1 University Hospital of Halle, Department of Internal Medicine IV, Hematology and Oncology, Halle (Saale), Germany 1

Introduction: Small nucleolar RNAs (snoRNAs) belong to a group of non-protein-coding RNAs with distinct functions in ribosomal RNA (rRNA) modification. snoRNAs are divided into two major classes: box C/D and box H/ACA snoRNAs. SnoRNAs assembly with small nucleolar ribonucleoproteins (RNPs) and guide those snoRNPs to the rRNA, where they catalyze rRNA modifications. Recent studies have shown that snoRNAs are dysregulated in multiple types of malignances. The functional role of snoRNAs in leukemia is unknown. Methods: We analyzed AML1-ETO induced snoRNA expression with snoRNA-seq of lin- murine bone marrow cells transduced with either AML1-ETO9a or with empty vector. Additionally we designed a GeCKO Crispr/Cas9 library with 1500 gRNAs targeting all known snoRNAs, snoRNA hostgenes as well as positive and negative controls. The library was infected with a MOI of 0.3 into Kasumi-1 cells. Cells were collected at day1, day5 and day9 after infection. Genomic DNA was extracted, PCR amplified for the integrated gRNAs and sequenced on a Miseq. Next generation sequencing data were analyzed by MAGeCK software. For functional analysis of single snoRNAs, we performed single knockouts of 2 snoRNAs in Kasumi-1 and MV4-11 cells using CRISPR/Cas9 technology. To analyze the effect of snoRNA knockout on colony formation ability of mutated cells, we embedded 300 cells/well in methylcellulose. Results: In the loss of function screen we identified SNORD14D and SNORD35A as most interesting candidates for growth of leukemia cells. Deep sequencing of snoRNAs in AML1-ETO9a transduced lin- murine bone marrow cells revealed significantly increased expression for snorRNAs. Interestingly SNORD14D and SNORD35A were also enriched. With colony-forming unit assays we confirmed our findings from the GeCKO screen. There was an up to 40% decreased clonogenic growth for Kasumi-1 and MV4-11cells bearing a CRISPR/Cas9 mediated knockout of SNORD14D or SNORD35A. Molecular analysis of altered ribosomal modifications demonstrated that knockout of SNORD35A in Kasumi-1 cells results in a significant reduction of 2-O-methylation on its specific rRNA modification side (28S C4506). For knockout of SNORD14D we could not detect a difference in 2-O-methylation of the specific side(18S C462), which may reflect rRNA modification independent functions of snoRNAs.

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Conclusion: Our data showed a functional relevance of snoRNAs for cell growth and proliferation of leukemia cells. Disclosure: No conflict of interest disclosed. P495

ABR, a novel inducer of transcription factor C/EBPα, is necessary for myeloid differentiation and a favorable prognostic factor in acute myeloid leukemia Namasu C.Y.1, Wurm A.A.1, Bräuer-Hartmann D.1, Hartmann J.-U.1, Katzerke C.1, Gerloff D.1, Hilger N.2, Fricke S.2, Schwind S.1, Christopeit M.3, Niederwieser D.1, Behre G.1 Leipzig University Hospital, Division of Hematology and Oncology, Leipzig, Germany, 2Fraunhofer Institute for Cell Therapy and Immunology, Germany, Leipzig, Germany, 3University Medical Center Hamburg-Eppendorf, Department of Stem Cell Transplantation, Hamburg, Germany 1

Introduction: Active BCR related (ABR) protein is closely homologous to BCR, which acts as a tumor suppressor in leukemia. Repression of ABR expression has been reported in t(8;21) acute myeloid leukemia (AML). However, the specific function of ABR in myeloid differentiation or AML has not been studied yet. Methods: We utilized acute myeloid leukemia U937 cell line, mouse bone marrow cells and human bone marrow cells from patients with acute myeloid leukemia. Cell differentiation was assessed by flow cytometry. Cell-cycle analysis was performed with propidium iodide and analyzed by flow cytometry. RNA detection was performed by quantitative real-time PCR. Protein detection was performed by immunoblot analyses. Transfections of constructs pEYFP-ABR, pcDNA3.1-E2F1, ABR siRNA or CEBPA siRNA in U937 cells were performed via electroporation. Results: We observed highly reduced ABR mRNA in AML patients from different karyotypes. Moreover, AML patients with high ABR expression survive significantly longer after hematopoietic stem cell transplantation. Here we found for the first time that older patients with AML that have high ABR expression benefited from azacytidine treatment, presenting an early response. Furthermore, treatment of leukemic cells with azacytidine induces ABR expression. ABR expression is increased while M-CSF and G-CSF stimulated myeloid differentiation of mouse bone marrow cells. In contrast to this, siRNA mediated block of ABR prevents myeloid differentiation. We identified ABR as a novel player in myelopoiesis via increasing the expression of the myeloid transcription factor C/EBPα, a major regulator of myeloid differentiation and functionally impaired in leukemia. Finally, ABR blocks cell-cycle progression and downregulates the cell-cycle activator E2F1, indicating the functional role of ABR as tumor suppressor in leukemic cells. Conclusions: Taken together, our data demonstrate that ABR plays a critical role in myelopoiesis via inducing C/EBPα and indicate the strong tumor suppressor potential of ABR expression in AML. Targeted treatments that increase endogenous levels of ABR might represent novel therapeutic strategies. Disclosure: No conflict of interest disclosed. P496

Longer observation times reveal long-latency leukemia induction from human AML cells engrafted into NSG mice Paczulla A.1, Dirnhofer S.2, Medinger M.3, Salih H.R.4, Rothfelder K.4, Passweg J.3, Tsakiris D.5, Lundberg P.5, Lengerke C.1,3 University Hospital Basel, Department of Biomedicine, Basel, Switzerland, University Hospital Basel, Department for Pathology, Basel, Switzerland, University Hospital Basel, Clinic for Hematology, Basel, Switzerland, 4University of Tuebingen Medical Center II, Department of Hematology & Oncology, Tuebingen, Germany, 5University Hospital Basel, Diagnostic Hematology, Basel, Switzerland 1 2 3

Introduction: Repopulation of immunodeficient mice remains the primary method to functionally assess human acute myeloid leukemia

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(AML) cells. Published data report engraftment of ~40 to 66% of AML (Eppert et al, 2011; Sanchez et al, 2009) depending on AML subtype and procedure, but mostly confined to intermediate or poor risk disease categories. Follow-up in these studies is however limited to 16 weeks. Here, we transplanted a mixed cohort of 18 primary AML samples and report that prolonging observation times allows robust engraftment and symptomatic leukemia induction from nearly every sample (~94%), including favorable risk molecular subtypes (e.g. inv(16)). Methods: 1×106 primary blasts purified by MACS technology from the peripheral blood (PB) of 18 patients with AML of different molecular backgrounds were mostly transplanted intravenously into the tail vein of sublethally irradiated NOD/SCID/IL2Rγnull (NSG) mice. Mice were monitored for disease signs and human engraftment via routine flow cytometry of PB and/or bone marrow (BM). Experiments were terminated at sickness or >1% human engraftment. Mice were analysed by whole body histopathology and multi-parameter flow cytometry of PB, BM and organs. Purified AML blasts were analysed by next generation sequencing (NGS). Results: At 16 weeks, 7 out of 18 (38%) samples showed engraftment. Strikingly however, engraftment was observed at a median time of 5.48 ± 2.34 months and in 56% of cases would have been missed using published protocols. Engraftment was confirmed by multi-parameter flow cytometry and whole body histopathology recognizing human leukemic cells in murine peripheral blood, bone marrow and peripheral organs. Next generation sequencing (NGS) retrieved a similar mutational profile in animal-derived versus pre-transplant samples. Interestingly, prolonging observation times only facilitated engraftment from CD34+ (but not CD34-) leukemic blasts, which molecularly showed enriched expression of stem cell genes. Conclusion: In summary, we show that leukemia engraftment can occur after prolonged latency and from (favorable risk) AML subtypes previously considered non-engraftable. This model may be used for in vivo studies on AML cell dormancy and disease initiation. Disclosure: No conflict of interest disclosed. P497

CD9 surface expression marks human MLL rearranged cells generated by genome editing Schneidawind C.1, Jeong J.2, Schneidawind D.1, Breese E.H.3, Porteus M.4, Kanz L.1, Cleary M.L.2 Department of Hematology and Oncology, Eberhard Karls University, Tuebingen, Germany, 2Department of Pathology, Stanford University, Stanford, United States, 3Department of Pediatrics, Cincinnati Children’s Hospital Medical Center, Cincinnati, United States, 4Division of Stem Cell Transplantation and Regenerative Medicine, Department of Pediatrics, Stanford University, Stanford, United States 1

Introduction: Genome editing provides a powerful approach to model the de novo development of human leukemias in primary human hematopoietic stem and progenitor cells (HSPCs) through targeted DNA double-strand breaks to generate chromosomal translocations. However, the very low abundance of appropriately edited cells bearing translocations and lack of markers for their identification or enrichment serve as barriers to their characterization and model development. Methods: Here we utilized genome editing to generate t(9;11) chromosomal translocations encoding MLL-AF9 and reciprocal AF9-MLL fusion products in CD34+ human cord blood cells. Results: Using a cytokine milieu tailored to the requirements of transformed cells we obtained selective monoclonal outgrowth in vitro of translocated cells comparable to the development of patient leukemia. This allowed a detailed characterization of genetically edited cells, which were distinguished by elevated MLL target gene expression and high surface CD9 expression in contrast to control cells. Conclusions: Defined cytokine milieus can overcome the limitations of obtaining homogeneous populations of initially rare translocated cells generated by genome editing techniques in primary human HSPCs. Furthermore, CD9 is a unique surface marker protein for detection and en-

Abstracts

richment of MLL rearranged cells, and may also serve as an option for targeted therapy. Disclosure: No conflict of interest disclosed. P498

CD97 expression mediates migration and adhesion of acute myeloid leukemia cells and modulates the bone marrow stromal microenvironment Wobus M.1, Jacobi A.2, Heidel K.1, Oelschlägel U.1, Bornhäuser M.1 Medizinische Klinik und Poliklinik 1, Medizinische Fakultät Carl Gustav Carus, TU Dresden, Dresden, Germany, 2Biotechnologisches Zentrum, TU Dresden, Dresden, Germany 1

Introduction: Bone marrow niches are specialized microenvironments that facilitate homing and survival of normal hematopoietic stem and progenitor cells (HSPCs) as well as leukemic stem cells (LCSs). Targeting the niche is a new strategy to eliminate persistent and drug-resistant LSCs. However, defined mechanisms mediating interactions of acute myeloid leukemia (AML) cells with the microenvironment remain largely unexplored. We have recently demonstrated that expression of the adhesion GPCR CD97 is elevated in blasts of FLT3-ITD positive AML patients. Here, we investigated the underlying mechanisms in more detail and the impact on mesenchymal stromal cells (MSCs) as a main cellular component of the bone marrow niche in vitro. Methods: FLT3-ITD MV4-11 and FLT3 wildtype OCI-AML3 AML cells were used for the in vitro experiments. CD97 knock-down was achieved by lentiviral transduction of plko1.6/shRNACD97. Primary MSCs were isolated from bone marrow aspirates of healthy donors or AML patients, co-cultured with AML cell lines in direct or indirect manner and were analyzed by flow cytometry and Western blot. Trans-well migration was analysed in a Boyden chamber assay. Results: CD97 knock-down by lentiviral transduction of plko1.6/shRNACD97 in MV4-11 leukemic cells caused inhibited trans-well migration towards FCS and LPA which is at least in part Rho-ROCK pathway dependent. Adhesion to a stromal layer was significantly decreased within two days and immunoblotting revealed inhibited Akt phosphorylation in the CD97 knock-down cells. The expression of the lysophosphatidic acid (LPA) receptor correlated with CD97 levels.In the second part of the study, we tested the impact of leukemic cells and their CD97 expression on the MSC phenotype. FACS analysis was performed after three days of MSC incubation with leukemic cell lines or their conditioned medium, respectively. Interestingly, CD90 and CD146 expression levels were increased by about 50% after coculture with MV4-11 wildtype cells but declined to the basic level after incubation with CD97 knock-down cells. In contrast, the expression levels of CD73 were increased by MV4-11 and even further elevated by CD97 knock-down cells. Comparable results were observed after MSC culture with conditioned medium of MV4-11 cells. Conclusion: In summary, our data suggest a modulation of the bone marrow microenvironment by leukemic cells expressing high CD97 levels rendering it a potential new theragnostic target. Disclosure: No conflict of interest disclosed. P499

Evasion of necroptosis and inflammasome activation promotes myeloid leukemogenesis Höckendorf U.1 Klinikum rechts der Isar der Technischen Universität München, III. Medizinische Klinik, München, Germany 1

Acute myeloid leukemia (AML) is partly driven by the loss of differentiation and the blockade of coordinated cell death of hematopoietic progenitors. As many hematopoietic disorders are characterized by the overproduction of pro-inflammatory cytokines, we hypothesized that necroptosis controlled cytokine secretion and inflammatory cell death might influence AML development.

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Here we show that RIPK3 limits oncogene-mediated leukemogenesis by promoting the inflammatory cell death of common myeloid progenitors (CMP) in experimental mice. Upon oncogenic stress RIPK3-dependent necroptosis and subsequent inflammasome activation were triggered, promoting the production of IL-1β, a potent stimulator of HSPC differentiation and maturation. Accordingly, gene-targeted mice deficient for necroptotic regulators or critical inflammasome components experienced a significantly accelerated AML development. Together, our data demonstrate RIPK3-mediated necroptotic cell death and inflammasome activation as a novel and unexpected tumor-suppressive mechanism in acute myeloid leukemia. Disclosure: No conflict of interest disclosed. P500

Suppression of c-Myc by PAM pathway inhibition and ATRA treatment for therapeutic gain in acute myeloid leukemia Schenk T.1,2, Stengel S.3, Kahl M.2, Rodriguez A.4, Perez A.4, Swords R.T.4, Petrie K.5, Zelent A.4 Klinik für Innere Medizin II / Universitätsklinikum Jena, Hämatologie und Internistische Onkologie, Jena, Germany, 2Zentrum für Molekulare Biomedizin (CMB), Institut für Molekulare Zellbiologie, Jena, Germany, 3Klinik für Innere Medizin IV / Universitätsklinikum Jena, Jena, Germany, 4University of Miami, Sylvester Comprehensive Cancer Center, Leukemia Program, Miami, United States, 5University of Stirling, Stirling, United Kingdom 1

Introduction: Defects in the proto-oncogene c-Myc have been widely implicated in the initiation and maintenance of acute myeloid leukemia (AML), however, c-Myc has been an elusive therapeutic target for drug development. In this study, we sought to target aberrant c-Myc expression by inhibition of the PAM signaling pathway in conjunction with all-trans retinoic acid (ATRA) as a strategy to reduce stem cell potential as well as to induce growth arrest, differentiation and apoptosis in AML cells. Methods: Small molecule inhibitors of PI3K (ZSTK474), mTOR (WYE125132) and PI3K/mTOR (NVP-BEZ235, dactolisib) where used alone or in combination with ATRA to treat AML cell lines and primary patient samples. Colony formation potential, cell growth, cell cycle arrest, differentiation and apoptosis were analyzed. c-Myc levels were monitored by qPCR and immunoblotting. Global gene expression profiling was conducted using Affymetrix GeneChip® ST Arrays. Results: PI3K and mTOR inhibition by ZSTK474 or WYE-125132 respectively, induced cell cycle arrest at low nanomolar concentrations in AML cell lines and primary patient samples. The dual PI3K/mTOR inhibitor NVP-BEZ235 was the most potent PAM inhibitor tested. Addition of ATRA further augmented the anti-leukemic effects of PAM inhibitors in vitro. Co-treatment lead to reduced colony formation capacity as well as cell cycle arrest, differentiation and programmed cell death. We observed a rapid down regulation of c-Myc following treatment with NVPBEZ235 via impaired translation, destabilization and enhanced degradation. ATRA treatment caused transcriptional and translational repression. NVP-BEZ235 combined with ATRA produced maximal c-Myc suppression and the greatest anti-leukemic effects. Detailed analysis of transcriptome changes in MV4-11 cells following treatment with NVP-BEZ235 and ATRA, revealed that both agents regulated the same biologic pathways, but acted on different gene sets within these pathways. Conclusions: Here we present a novel multi-target approach to suppress aberrant c-Myc levels in AML. While inhibition of the PAM pathway leads to inhibition of c-Myc translation, destabilization and degradation, supplementary treatment with ATRA can further decrease c-Myc levels by inhibition of its transcription as well as translation. These data support the clinical investigation of ATRA combined with PAM inhibitors especially in AML patient with high c-Myc activity. Disclosure: No conflict of interest disclosed.

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P501

Analysis of snoRNA expression in acute myeloid leukemia Zhou F.1, Pauli C.1, Gerloff D.1, Rohde C.1, Köhn M.2, Misiak D.2, Hüttelmaier S.2, Müller-Tidow C.1 Universitätsklinikum Halle, Universitätsklinik und Poliklinik für Innere Medizin IV, Halle, Germany, 2Institut für Molekulare Medizin, Sektion Molekulare Zellbiologie, Martin-Luther-Universität, Halle, Germany 1

Introduction: Small nucleolar RNAs (snoRNAs) belong to the group of non-protein-coding RNAs with distinct functions in ribosomal RNA (rRNA) modification. snoRNAs are divided into the two major classes: box C/D and box H/ACA snoRNAs. SnoRNAs assembly with small nucleolar ribonucleoproteins (RNPs) and guide those snoRNPs to the rRNA, where they catalyze rRNA modifications. C/D box snoRNAs are involved in 2-O-methylation, whereas H/ACA snoRNAs guide pseudouridylation. Recent studies showed that snoRNAs play a role in cancerogenesis. Hence, we analyzed snoRNAs expression and function in leukemogenesis. Methods: To understand the role of snoRNAs in leukemogenesis, we analyzed the snoRNA expression pattern of 63 AML patients, registered in a recent clinical trial (ClinicalTrials.gov NCT00915252). Additionally we analyzed snoRNA expression pattern in AML1-ETO or Myc transduced lin- mouse bone marrow cells. RNA was isolated with Qiagen miRNeasy Kit. Librarys for small RNA (40–200 nt) specific next generation sequencing (NGS) where prepared TruSeq Small RNASample Prep Kit and size separation of 40–200 nucleotides. NGS was performed on an Illumina HiScanSQ, using 50 cycles of single read sequencing. Results: Next generation sequencing data revealed 229 C/D box snoRNAs and 90 H/ACA box snoRNAs were expressed in one or more patient samples. In further analysis we compared clinical data and snoRNA expression pattern. Here we could show that snoRNA expression clustered with specific risk groups. High levels of snoRNA expression associated with intermediate molecular risk (269 snoRNAs, p ≤ 0.05) and a poor response to chemotherapy (101 snoRNAs, p ≤ 0.05). Further, we could show that snoRNAs are higher expressed in AML patient samples with NPM1 wildtype (124 snoRNAs, p p ≤ 0.05). Transduction of the oncogenes AML1ETO9a or Myc to normal lin- mouse bone marrow cells induces an increased overall snoRNA expression. To understand the role of snoRNAs during myeloid differentiation, we stimulated HL60 cells with ATRA (1µM) for 72h and analyzed snoRNA expression by NGS. We found 93 snoRNAs downregulated (fold change ≤0.8) and 88 upregulated (fold change ≥1.2) during ATRA induced differentiation of HL60 cells. Conclusion: In summary our data identified an altered snoRNA expression profile in leukemogenesis and myeloid differentiation. Disclosure: No conflict of interest disclosed. P502

Identification and functional analysis of novel putative snoRNAs in acute myeloid leukemia Gerloff D.1, Rohde C.1, Zhou F.1, Pauli C.1, Köhn M.2, Misiak D.2, Hüttelmaier S.2, Müller-Tidow C.1 Universitätsklinikum Halle, Universitätsklinik und Poliklinik für Innere Medizin IV, Halle, Germany, 2Institut für Molekulare Medizin, Sektion Molekulare Zellbiologie, Martin-Luther-Universität, Halle, Germany 1

Introduction: Small nucleolar RNAs (snoRNAs) are a large group of non-coding RNAs, divided into two major families, box C/D and box H/ ACA snoRNAs, based of common sequence motifs and structural features. The snoRNAs guide small nucleolar ribonucleoproteins (snoRNPs) to complementary regions of ribosomal (rRNA) or small nuclear RNAs (snRNA), where the snoRNP complexes catalyze ribosomal modifications. Additionally snoRNAs affect alternative splicing, RNA decay and protein functions. Recent studies showed the relevance of snoRNAs for the pathogenesis of cancer. Methods: To understand the role of snoRNAs in leukemogenesis, we analyzed the snoRNA expression pattern of 63 AML patients, registered in a

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clinical trial (ClinicalTrials.gov NCT00915252). Librarys for small RNA specific next generation sequencing (NGS) where prepared with TruSeq Small RNASample Prep Kit and size separation of 40–200 nucleotides. NGS was performed on an Illumina HiScanSQ, using 50 cycles of single read sequencing. Sequencing data were processed with Cutadapt, mapped with Bowtie 2 to the human genome (hg19). Abundant regions were detected with Bedtools genomeCoverageBed and were annotated using snoRNABase (LBME), DASHR, UCSC snoRNA and repeatmasker track. For further analysis reads of discovered regions were calculated as reads per million (RPM). Regions without annotation were analyzed for C/D box motifs to discover putative C/D box snoRNAs. Results: Small RNA specific next generation sequencing revealed 1396 mapped regions, which could not be annotated as known small noncoding RNAs. In silico analysis identified 90 putative C/D box snoRNAs. All of these putative snoRNAs contain the box C/D motifs and were mostly located in introns of known genes. In further analysis we found that increased expression of putative snoRNAs was found in patients with an intermediate molecular risk (51 snoRNAs, p ≤ 0.05) and in patients with a bad response to chemotherapy (20 snoRNAs, p ≤ 0.05). To analyze the binding of 8 putative snoRNA candidates to the snRNP complex we performed NOP58 and FBL specific RNA immunoprecipitation (RIP) in Kasumi-1 cells. Here, we could show an enriched binding of 2 putative snoRNAs to FBL and NOP58. In functionall analysis we could show, that a CRISPR/Cas9 mediated knockdown of single putative snoRNAs influences colony formation and cell growth of Kasumi-1 and MV4;11 cells. Conclusion: We identified novel unknown snoRNAs, which contribute to neoplastic transformation. Disclosure: No conflict of interest disclosed. P503

Proapoptotic effects of Deferasirox on acute myeloid leukemia (AML) cell lines Hinterleitner C.1, Malenke E.1, Bugl S.1, Haap M.2, Müller M.R.1, Heißner K.1, Kopp H.-G.1 Department of Medicine II, Eberhard Karls University, Medical Oncology, Hematology, Immunology, Rheumatology and Pulmology, Tübingen, Germany, 2 Department of Internal Medicine, Eberhard Karls University, Medical Intensive Care Unit, Tübingen, Germany 1

Introduction: Anti-proliferative effects of deferasirox (DFX), an orally available iron chelator, in various cancer cell types have recently been described. DFX is sufficient to induce apoptosis in several acute myeloid leukemia (AML) cell lines by activating caspases 3/7 and 9. Because the underlying mechanisms remain unknown, subcellular effects of DFX on four human leukemia cell lines were studied in vitro. Methods: HL60, THP-1, U937 and MM6 cells were treated with DFX at variable doses in vitro. Cell proliferation and involvement of BCL-2 in apoptosis were analyzed by Cell Titer Glo assay. Cell cycle distribution, cell death, and apoptosis were evaluated by Caspase 3/7 Glo assay, immunoblotting, and flow cytometry. DNA damage and DNA damage response (DDR) were analyzed by immunocytochemistry detecting S139-posphorylation of histone H2AX (yH2AX) and 53BP1. Results: DFX decreased proliferation and viability in all myeloid cell lines in a dose- and time-dependent manner. In HL-60, THP-1, U937 and MM6 cells DFX induced G1/S phase cell-cycle arrest, which was followed by massive caspase-mediated apoptosis with profound activation of caspases 8, 9, 7, and 3 as well as poly(ADP-ribose) polymerase (PARP) cleavage. Interestingly, cotreatment with DFX and ABT-199, a potent and selective Bcl-2 inhibitor, showed highly synergistic effects in all tested cell lines. Remarkably, DFX induced apoptotic cell death was preceded by S139-posphorylation of histone H2AX (yH2AX), a prototypical surrogate marker of DNA double-strand breaks (DSBs) and induction of 53BP1. Conclusion: In all tested AML cell lines DFX leads to a massive accumulation of DSBs followed by G1/S phase cell-cycle arrest and consecutive, Bcl-2 dependent caspase-driven apoptosis. Our results provide new insights into the complex molecular mechanism of iron chelation in human

Abstracts

AML. We suggest that treatment with DFX alone or in combination with ABT-199 might have beneficial effects in future AML treatment. Disclosure: No conflict of interest disclosed. P504

Oncogenic splicing of the Apoptosis-Stimulating Protein of p53-2 (ASPP2) is an initiating step in leukemogenesis facilitating acquisition of structural genomic mutations Kampa-Schittenhelm K.1, Mau-Holzmann U.2, Lopez C.3, Schittenhelm M.1 Universitätsklinikum Tübingen, Medizinische Klinik II, Tübingen, Germany, Universitätsklinikum Tübingen, Medizinische Genetik, Tübingen, Germany, 3 Oregon Health and Science University, Dept. of Hematology and Oncology, Portland, United States 1 2

ASPP2 is an independent haploinsufficient tumor suppressor which initiates induction of apoptosis after cellular damage in a TP53 dependent manner. We recently reported the identification of two C-terminally truncated splice variants of ASPP2 with high frequency in acute leukemias. Tantalizingly, all variants lack the functionally relevant TP53 binding sites. The most prevalent isoform lacking exon 17 (ASPP2k) is detected in up to 60% of acute leukemias. A cell sort for the CD34+ stem cell fraction confirmed ASPP2k-expression in the leukemia-initiating clone. Immunoblotting revealed translation of ASPP2k into a genuine protein isoform. Generation of isoform-specific antibodies allows to discriminate ASPP2k from the wildtype isoforms: Interestingly, ASPP2k is the sole isoform expressed in some native leukemia samples. Other samples reveal a heterozygous expression pattern suggesting a dominant-negative function of the C-terminally truncated ASPP2k isoform. Forced expression of ASPP2k using a HisMax plasmid vector in murine IL3-dependent pro-B Ba/F3 cells induced a more aggressive phenotype with mitotic failure and perturbed cellular proliferation compared to the empty vector controls. Tantalizingly, IL3 was successfully weaned in the ASPP2k cells – indicating that autoactivating genomic alterations must have occurred upholding cellular integrity and viability. Fractionated gamma irradiation accelerated this process. In contrast, the empty vector strains did not survive IL3 weaning. Chromosomal analyses of IL3-independent Ba/F3 ASPP2k cells revealed structural alterations, including monosomies and additional marker chromosomes. Furthermore, we demonstrate that ASPP2k is stress-inducible. Changing culture conditions (e.g. temperature) of cell lines or ex vivo native AML samples resulted in a dramatic increase of the ASPP2k isoform. Consecutive treatment of cells with daunorubicin lead to relative resistance of pre-stressed cells compared to their counterparts cultured under standard conditions. Vice versa, specific knock-down of ASPP2k resulted in a significant increase in induction of apoptosis upon chemotherapy compared to the empty vector controls. Our data demonstrates that ASPP2k plays a distinctive role as an anti-apoptotic regulator of the TP53 checkpoint, rendering cells to a more aggressive phenotype as evidenced by proliferation and apoptosis rates – and facilitates acquisition of genomic mutations, a first initiating step in leukemogenesis. Disclosure: Kerstin Kampa-Schittenhelm: Honoraria: ja Marcus Schittenhelm: Honoraria: ja P505

The PDK1-AKT axis in acute leukemia – A potential resistance mechanism towards receptor tyrosine kinase (RTK) targeted therapies Kampa-Schittenhelm K.1, Haug N.1, Kanz L.1, Schittenhelm M.1 Universitätsklinikum Tübingen, Medizinische Klinik II, Tübingen, Germany

Constitutive activation of the AKT pathway, e.g. via mutant RTKs, is a central observation in acute leukemias. Consequently, forced expression of AKT results in development of leukemia in mouse models. Tyrosine kinase inhibitors insufficiently inhibit the AKT pathway, leaving the PDK1-

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AKT axis unaffected. We aim to determine the role of PDK-1 in acute leukemia, which may provide an escape mechanism towards TKI. To determine the role of PDK1 in acute leukemia, we established different PDK knock-out (ko) leukemia cell models using a lentiviral shRNA approach. Ko and empty vector control cells were treated with standard chemotherapeutics (eg. daunorubicin) as well as specific tyrosine kinase inhibitors (TKI) and induction of apoptosis and proliferation rates were assessed. Signal transduction analysis by (phospho)proteome analyses, immunoblotting, intracellular phospho-peptide specific flow cytometry and human phospho-kinase profiler arrays were performed +/- different TKI (dasatinib, sunitinib, imatinib). FLT3, KIT or BCR/ABL-inhibition does not sufficiently globally silence AKT signaling in MOLM14, Kasumi1 and K562 acute leukemia cells as (i) threonine phosphorylation of AKT at codon 308 remained unaffected and (ii) downstream AKT signaling remained activated. Knock-down of PDK1 resulted in consecutive de-phosphorylation at AKT(Thr308), the target site of PDK-1, and p70S6Kinase downstream signaling. Global phospho-kinase protein profiling revealed 378 significantly differently regulated phosphorylation sites of the ko strains compared to the empty vector controls. Interestingly, among the highest regulated genes were several genes involved in cell cycle regulation (e.g. pRB and cyclin D). Differences in cell cycle regulation of PDK1 expressing and ko cells were verified by cell cycle analyses and Western immunoblotting. Consequently, on the cellular level, PDK-1 interference resulted in slower cellular proliferation rates compared to the empty vector controls. Exposure of cells towards chemotherapeutics (such as daunorubicin) increased the proapoptotic cell fraction in the ko strains. Conclusion: Taken together, our data suggest that PDK1-AKT signal transduction contributes to aggressiveness and therapy susceptability of the leukemic clone. Targeting the PDK-1-AKT axis may provide a therapeutic strategy to overcome therapy resistance. Disclosure: No conflict of interest disclosed. P506

The role of attenuated Apoptosis Stimulating Protein of p53-1 (ASPP1) in acute myeloid leukemia (AML) Kampa-Schittenhelm K.1, Kaiser M.1, Blumenstock G.2, Schittenhelm M.1 Universitätsklinikum Tübingen, Medizinische Klinik II, Tübingen, Germany, Universitätsklinikum Tübingen, Klinische Epidemiologie, Tübingen, Germany

1 2

ASPP1 belongs to a family of p53-binding proteins and enhances apoptosis by stimulation of p53-transactivation of selected proapoptotic target genes. It is preferentially expressed in hematopoietic stem cells (HSC) and together with p53 preserves the genomic integrity of the HSC pool. Consequently, attenuated expression of ASPP1, which is linked to methylation of the promoter region, has been associated with malignant transformation and development of acute lymphoblastic leukemia and lymphomas. We now provide evidence that ASPP1 is highly altered in AML, suggesting a role in leukemogenesis as well as therapy response. ASPP1 mRNA and protein expression levels of freshly isolated native patient samples (47) and healthy bone marrow donors (17) were determined by qRT-PCR and Western immunoblotting. Statistical analyses were performed. To explore implications of attenuated ASPP1 levels with regard to apoptosis induction and proliferation, ASPP1-expressing leukemia cell lines (MOLM14, HL60, Jurkat) or native patient samples were stably silenced using a retroviral shRNA approach. XTT viability and annexin V-based apoptosis assays were performed using standard chemotherapeutics in comparison to empty vector controls. Decitabine was used as an epigenetic sensitizer via hypomethylation of the promoter region. ASPP1 mRNA expression was found to be frequently and highly statistically significantly (p = 0.001) attenuated in AML. Low ASPP1 mRNA levels thereby translated into attenuated protein expression. Retroviral ASPP1-interference lead to perturbed proliferation capacities (up to 3-fold increase) and attenuated apoptosis upon standard chemotherapeutics in leukemia cell lines as well as native leukemia blasts.

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Intriguingly, epigenetic therapy using the hypomethylating agent decitabine resulted in upregulation of ASPP1 expression in leukemia cells with low basal ASPP1 levels. Consequently, decitabine pretreatment sensitized these patient samples towards chemotherapy with a favorable proapoptotic overall efficacy compared to chemotherapy alone. Our results demonstrate that dysfunctional regulation of ASPP1 expression likely contributes to the biology of leukemogenesis and to primary therapy resistance in a subgroup of patients with acute leukemia and seems to be linked to hypermethylation. Prospective clinical studies are warranted to evaluate the role as a biomarker for risk stratification in leukemia patients and for monitoring therapy responses. Disclosure: No conflict of interest disclosed. P507

Upregulation of the anti-apoptotic iASPP (inhibitory Apoptosis Stimulating Protein of p53) is involved in therapy resistance and inferior outcome in acute myeloid leukemia (AML) Bajrami Saipi M.1, Schittenhelm M.1, Blumenstock G.2, KampaSchittenhelm K.1 Universitätsklinikum Tübingen, Medizinische Klinik II, Tübingen, Germany, Universitätsklinikum Tübingen, Klinische Epidemiologie, Tübingen, Germany

1 2

Inactivation of the p53 pathway is a frequent event in human cancers promoting tumorigenesis and resistance to chemotherapy. Inactivating p53 mutations are uncommon in non-complex karyotype leukemias suggesting that the p53 pathway must be inactivated by other mechanisms. ASPP proteins are a family of p53-binding proteins that consists of three members: the pro-apoptotic ASPP1 and ASPP2 and the apoptosis-inhibiting iASPP. All play a crucial role in the regulation of p53-induced apoptosis. We have previously demonstrated that the proapoptotic family member ASPP2 is significantly attenuated in AML – and this uniquely links to the patient cohort failing induction chemotherapy. We have now studied the role of iASPP in acute myeloid leukemia. iASPP mRNA levels were determined using standard qRT-PCR. Protein expression was measured using flow cytometry and immunoblot assays. Basal iASPP levels of 95 leukemia patients in bone marrow as well as in peripheral blood samples were normalized to a control cohort comprising 24 healthy bone marrow donor samples. Blood samples were obtained before and during initial therapy at defined timepoints and expression levels of iASPP were correlated with blast count and initial therapy response. Further, leukemia cell lines stably suppressing iASPP were generated using a lentiviral-based shRNA approach. Changes in proapoptotic activity and proliferation were analyzed in iASPP knock-out cells versus empty vector controls. We confirmed statistically significantly higher basal iASPP mRNA as well as protein levels in AML samples compared to the donor controls. Decreasing iASPP levels under induction therapy positively correlated with the response to therapy. Interestingly, highest iASPP levels were found in patients expressing low ASPP2 levels – and this correlated with poor clinical outcome. These in vivo data were backed up in our cell models: as expected, knockdown of iASPP in different cell lines as well as in ex vivo blasts resulted in significantly increased apoptosis upon treatment with standard chemotherapy (e.g. daunorubicin). In line, proliferation time slowed down in the ko cells compared to the empty vector controls. We believe that different members of the ASPP family play a distinct role in acute leukemia and that complex interactions between the different family members are in part responsible and predictive for therapy outcome. Disclosure: No conflict of interest disclosed.

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Loss of DNMT3A function prevents reporter gene inactivation in leukemic cell lines and mES cells Schneider D.M.1,2, Beckert J.1,2, Ehrenfeld S.1,2, Khan R.1, Mitschke J.1,2, Shoumariyeh K.1, Veratti P.1,2, Miething C.1,2 Uniklinik Freiburg, Freiburg, Germany, 2DKFZ Heidelberg, DKTK, Freiburg, Germany 1

More than 70% of de novo AML cases harbor at least one amino acid change in a DNA methylation-related gene or epigenetic modifier. Up to 30% of normal karyotype AML cases show a somatic mutation in DNA (-cytosine-5)-methyltransferase 3 alpha (DNMT3A). These mutations are mostly heterozygous and more than half of the DNMT3A mutations alter arginine 882 in its catalytic domain (mostly into histidine). This mutation has been shown to dominantly inhibit DNMT3Awt by blocking its homo-tetramerization. The importance of DNMT3A for the differentiation of hematopoietic stem cells (HSCs) is demonstrated by the fact that DNMT3A-KO HSCs show increased self-renewal and reduced differentiation. DNMT3A mutations are assumed to arise early in AML development, most probably in HSCs, so that a clonally expanded pool of pre-leukemic cells arises from which AML evolves. As DNMT3A is highly conserved, we explored the effect of DNMT3A mutation in leukemogenesis via its knock-down and over-expression in human AML cell lines and murine embryonic stem (mES) cells. The knock-down was achieved using a two colored, antibiotic selectable and tet-inducible retroviral shRNA expression vector system. Murine ES cells carrying a “homing cassette” were targeted via recombination-mediated cassette exchange to contain our gene of interest, a fluorescence marker and an antibiotic selection resistance gene. Furthermore, to analyze the role of DNMT3A in tumor maintenance, bone marrow transplantation experiments with FLt3-ITD and inducible DNMT3Awt or R882H and a shRNA targeting murine DNMT3A are ongoing. Analyzing the DNMT3A-inducible cell lines, we found that over-expression of DNMT3A wt increases the inactivation of retroviral reporter constructs via promoter methylation, both in the leukemia cell lines HL60 and K562 as well as in murine embryonic stem cells. In contrast, the R882H mutation reduced the effect of DNMT3A over-expression in HL60 and murine ES cells, but less so in K562 cells. DNMT3A knock-down was significantly selected against in the different leukemic cell lines, suggesting residual functionality of DNMT3A is required. Disclosure: No conflict of interest disclosed.

ine-kinase inhibitors. However, the regulating mechanisms and the role of antioxidants of ROS-dependent regulation of proliferation are still unclear. Methods: To study the functional role of ROS in JAK2-VF positive cells, granulocytes from peripheral blood of VF-positive patients and healthy controls were isolated and analyzed for ROS level (H2DCFDA method), activity of SOD1/2, catalase and content of glutathione, respectively. Granulocytes were stimulated with cytokines for 1h, including IL-3 (10ng/ml), IL-6 (10ng/ml), TNFa (1ng/ml) and Epo (3IU/ml) followed by analysis of ROS formation. To investigate the role of the antioxidants NAC (5–25mM, 24h) and Tempol (0.25–5mM, 24h) in proliferation (MTT assay) and in cell cycle control (PI stain, immunoblot), we used murine promyeloid cell lines 32D JAK2-WT and JAK2-VF. Furthermore, we studied the effects of a combination of both antioxidants and Ruxolitinib (Rux, 100–500nM, 24h) for induction of apoptosis (AnnexinV, immunoblot). Results: Granulocytes of VF-positive patients showed higher steady state ROS levels, as well as higher activities of SOD1/2 and reduced activity of catalase. Stimulation of VF-positive granulocytes with cytokines for 1h led to rapid and enhanced ROS induction as compared to healthy controls. Investigating the role of intracellular ROS for proliferation and cell cycle progression, 32D JAK2-VF cells were incubated with NAC and Tempol for 24h resulting in reduction of ROS levels, proliferation and inhibition of G1-to-S phase transition likely caused by reduction of protein expression of CyclinD1/D2, CyclinE and induction of p27 and p19. Furthermore, combined treatment with Rux/NAC and Rux/Tempol strongly increased apoptosis after 24h paralleled by reduced expression of Bcl-xl and Mcl-1. Conclusion: We here show, that JAK2-VF-positive cells are characterized by dysbalanced ROS formation and enhanced cytokine-dependent ROS-induction. In addition, we could show that reduction of ROS by antioxidants may offer a novel therapeutic option in treatment of VF-positive patients. Disclosure: No conflict of interest disclosed. P510

Expression level of JAK2-V617F determines sensitivity to tyrosine kinase inhibitor Ruxolitinib in HEK293 Flp-In cells and can be augmented by treatment with inhibitors of PI3K/ Akt pathway in JAK2-V617F – positive murine progenitor cells Müller C.1, Schnöder T.2, Edelmann-Stephan B.1, Hessenkemper W.3, Bongartz H.3, Schaper F.3, Fischer T.1 Department of Hematology and Oncology, Otto-von-Guericke University, Magdeburg, Germany, 2Medical Faculty, Internal Medicine II, Friedrich-SchillerUniversity, Jena, Germany, 3Department of Systems Biology, Otto-von-Guericke University, Magdeburg, Germany 1

Posterdiskussion

Myeloproliferative Neoplasien, Gerinnung

Introduction: Activating mutations within protein kinases, e.g. JAK2V617F (JAK2-VF) and Bcr/Abl, are characterized by high intracellular levels of reactive oxygen species (ROS) mediated by constitutive activation of pathways controlling ROS production. It has been supposed, that imbalance of these signaling pathways may lead to disturbed proliferation, aggressive biological phenotype and resistance to treatment with tyros-

Introduction: Chronic myeloproliferative neoplasms are characterized by acquired somatic mutation V617F of the JAK2-gene (JAK2-V617F, JAK2VF) in 95% of polycythemia vera and 50% of essential thrombocytosis and primary myelofibrosis patients. This mutation leads to constitutive activation of several signaling pathways (e.g. STATs, PI3K/Akt, MAPKs, PLCg1). Treatment of JAK2-V617F – positive patients with Ruxolitinib (Rux) could improve clinical symptoms, e.g. reduction of spleen size, but failed to eradicate the malignant clone. However, the role of JAK2-V617F allelic ratio for sensitivity to Rux treatment is still unknown. Methods: To simulate the effects of different expression levels of JAK2V617F, we used HEK293 Flp-In stable transfectans inducibly expressing JAK2-WT and JAK2-V617F for titrating different expression levels and characterized their behavior to activation of JAK2-related proteins (immunoblot), calcium flux (Indo-1) and sensitivity to Rux treatment (0.25–1.0µM, 12h; AnnexinV, immunoblot) 12h after induction of protein expression by doxycycline. To characterize synergistic effects on apoptosis of Rux with inhibitors for PI3K/Akt (LY294002 2.5–12.5µM; Akt inh. VIII 0.25–1.0µM) and MEK (U0126 1.0–5.0µM), we used the murine progenitor cell line 32D JAK2-VF (AnnexinV, TMRE, mitochondrial swelling). Results: In HEK293 Flp-In transfectans, we could induce three different expression levels of JAK2-VF compared to WT controls by doxy-

Abstracts

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P509

Regulatory role of intracellular ROS levels and effects of antioxidants (NAC, Tempol) on proliferation, cell cycle and sensitivity to treatment with tyrosine kinase inhibitor Ruxolitinib in JAK2-V617F – positive myeloproliferative neoplasms Müller C.1, Schnöder T.2, Edelmann-Stephan B.1, Wolleschak D.1, Simeoni L.3, Fischer T.1 Department of Hematology and Oncology, Otto-von-Guericke University, Magdeburg, Germany, 2Medical Faculty, Internal Medicine II, Friedrich-SchillerUniversity, Jena, Germany, 3Institute for Molecular and Clinical Immunology, Otto-von-Guericke University, Magdeburg, Germany 1

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cycline (0.5-fold, 1.0-fold, 2.0-fold of JAK2-WT expression). Increased JAK2-VF-expression resulted in enhanced activation of PLCg1, STAT5, Akt and Erk, as well as increased calcium levels. The induction of different JAK2-VF expression-levels also affected the response to Rux treatment likely by reduction of Bcl-xl and Mcl-1 expression and reduced induction of Bax. Furthermore, in 32D JAK2-VF cells the combined treatment with Rux/LY294002 and Rux/Akt inh. VIII showed a synergistic effect on induction of apoptosis. This was associated with reduction of mitochondrial potential, increased mitochondrial swelling and reduced expression of Bcl-xl and Mcl-1. Conclusion: Here we show that titrating of JAK2-VF expression level in HEK293 cells is accompanied by strong induction of JAK2-related signaling pathways and reduced sensitivity to Rux treatment. Furthermore, we could demonstrate, that combined treatment with Rux and inhibitors of PI3K/Akt pathway may offer novel strategies in treatment of VF-positive patients. Disclosure: No conflict of interest disclosed. P511

2016 Interim analysis of the prospective, non-interventional study JAKoMo: new insights into ruxolitinib treatment of myelofibrosis in daily clinical routine Geer T.1, Heinrich B.2, Jacobasch L.3, Tesch H.4, Wehmeyer J.5, Schlag R.6, Gröschl B.7, Markhauser M.7, Bachhuber P.7, Koschmieder S.8 Medizinische Klinik III, Diakonie-Klinikum Schwäbisch Hall, Schwäbisch Hall, Germany, 2Hämatologisch-onkologische Praxis Brudler/Heinrich/Bangerter, Augsburg, Germany, 3Onkologische Schwerpunktpraxis, Dresden, Germany, 4 Centrum für Hämatologie und Onkologie Bethanien, Frankfurt/ Main, Germany, 5Gemeinschaftspraxis Wehmeyer/ Lerchenmüller/ Kratz-Albers/ Timmer/ Bieker/ Liersch, Münster, Germany, 6ISCO GmbH Studiengesellschaft, Würzburg, Germany, 7Novartis Pharma GmbH, Nürnberg, Germany, 8Klinik für Hämatologie, Onkologie, Hämostaseologie und SZT, Med. Fakultät Uniklinik RWTH Aachen, Aachen, Germany 1

Introduction: Ruxolitinib (RUX) is a potent, selective JAK1/JAK2 inhibitor approved for the treatment of diseaserelated splenomegaly or symptoms in adult patients (pts) with primary myelofibrosis (PMF), post-polycythemia vera MF (PPV-MF), or post-essential thrombocythemia MF (PETMF). This prospective, noninterventional study (JAKoMo, CINC424ADE05) is designed to collect data regarding the current use and outcome of RUX therapy in daily clinical practice in Germany. Methods: A population of 379 JAK1 inhibitor- and JAK2 inhibitor-naive pts documented from 09/2012 until 02/2016 was analyzed. RUX was administered according to the summary of product characteristics (SmPC). Drug utilization, efficacy, safety and tolerability, quality of life and pharmacoeconomics were documented for each participating patient. Results: In this interim analysis, data of 379 pts from 91 German centers were analyzed, with a median treatment duration of 1.04 years (range 0.02–3.18 years). The majority of centers enrolled a small number of pts (68.2% of centers enrolled 4 pts or less) confirming the low incidence of MF. The median age of pts was 72.0 years and gender distribution was balanced (52.2% male). The most common diagnosis was PMF (64.9%), followed by PPV-MF (20.8%) and PET-MF (12.1%), with information on MPN subtype pending in 2.1%. For the majority of pts (58.6%), the IPSS score was not documented. Of the residual 157 pts, 10.8% had a low risk, 25.5% had an Int-1 risk, while 42.0% had an Int-2 risk and 21.7% had a high risk profile. The median palpable spleen length at baseline was 14.5 cm (range 0.0–29.0 cm; N = 159). 52.8% suffered from general weakness at baseline (most common documented symptom). The majority of pts (82.9%) showed an ECOG performance status of either 0 or 1, which was in keeping with a typical outpatient population. 74.7% of pts started therapy with a total daily dose of at least 20 mg RUX. After 6 months of therapy, 21.9% of subjects were still suffering from weakness (N = 74), the median palpable spleen length was 9.0 cm (N = 51). The most common AEs were anemia (30.1%; grade 3/4: 6.1%) and thrombocytopenia (28.5%; grade 3/4: 7.7%). Further efficacy and safety data will be presented at the conference.

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Conclusion: This interim analysis of the current JAKoMo non-interventional study depicts a representative group of MF pts treated with RUX and confirms the safety and efficacy profile obtained within the two COMFORT trials in daily clinical practice. Disclosure: Thomas Geer: Financing of Scientific Research: Honorar für Präsentation des Posters; Expert Testimony: Entschädigung für die Dokumentation von Daten innerhalb der nicht-interventionellen Studie JAKoMo Steffen Koschmieder: Expert Testimony: Entschädigung für die Dokumentation von Daten innerhalb der nicht-interventionellen Studie JAKoMo P512

Detection of free circulating c-KIT D816V in plasma using ddPCR to monitor disease activity in systemic mastocytosis Shoumariyeh K.1, Follo M.1, Philipp U.1, Duyster J.1, von Bubnoff N.1 Universitätsklinikum Freiburg, Medizinische Klinik I, Freiburg i. B., Germany

Introduction: In systemic mastocytosis (SM), serum tryptase levels correlate with mast cell burden but do not accurately predict disease activity or outcome. Most patients harbor a c-KIT D816V mutation. The c-KIT D816V allele burden measured by quantitative real-time PCR on bone marrow or whole blood samples recently was shown to correlate with SM subtype and to predict survival. We aimed to establish a sensitive assay for quantitative detection of c-KIT D816V using free plasma circulating (fc)DNA. Methods: We established a hydrolysis probe based, Locked Nucleic Acid assay to detect c-KIT D816V and wild type c-KIT ctDNA by droplet digital PCR on a BioRad QX100 system. Reaction sensitivity was 0.01% with high specificity. We analyzed 11 plasma samples from 3 patients (pts) with c-KIT D816V positive SM. Results were correlated with disease activity and tryptase levels. Results: The c-KIT D816V mutant was detected in all 3 pts in all plasma samples. The amount of c-KIT D816V fcDNA (copies/ml plasma) correlated with disease activity in each of the pts, though the absolute c-KIT D816V copy numbers differed between the pts (0,18–412,22). The first patient fulfilled criteria for aggressive systemic mastocytosis (ASM) with biopsy-proven involvement of the colon and C-signs (Malabsorption). The mean c-KIT D816V copies/ml plasma was 214 (range 92,98–412,22). The second patient had indolent systemic mastocytosis (ISM) with biopsy-proven involvement of the ileum and colon as well as a B-sign (persistent tryptase levels > 200 ng/ml). The mean c-KIT D816V copies/ml plasma for this patient was 39 (range 30,78–46,67). The third patient had systemic mastocytosis with associated chronic myelomonocytic leukemia (SM-AHNMD). The mean c-KIT D816V copies/ml plasma was 1,5 copies/ml plasma (range 0,18–4,57). In all 3 pts increase or decrease of c-KIT D816V ctDNA copy numbers in serial samples correlated with disease activity and response to cytoreductive treatment, including a persisting positive to negative conversion of the patient with AHNMD after treatment with 2CDA and Midostaurin (PKC412). Conclusion: Our preliminary results indicate that c-KIT D816V mutant fcDNA in pts with SM can be used as a disease-specific surrogate for disease activity and response to treatment. We therefore initiated a pilot trial to evaluate the significance of c-KIT D816V mutant fcDNA as a disease-specific biomarker in SM. Disclosure: No conflict of interest disclosed. P513

Molecular monitoring of major BCR-ABL1 isoform p210 by qPCR reveals differential response to tyrosine kinase inhibitors in accelerated phase CML: a case series Altahan R.1, Alsohaibani L.1, Khoja O.T.1, Zaidi S.1, Alghamdi M.1, Pukhta I.A.1, Tashkandi S.1, Peer Zada A.A.1 King Fahad Medical City, Riyadh, Saudi Arabia

Background: Chronic myeloid leukemia (CML) patients are characterized by the BCR-ABL1 fusion protein (p210) that leads to a constitutive pro-

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tein tyrosine kinase activity thereby giving a myeloproliferative phenotype in CML. CML occurs in three different forms: the chronic CP-CML, the accelerated AP-CML and the blast crisis BC-CML, each of which is clinically managed according to standard guidelines (ELN, NCCN) through the use of tyrosine kinase inhibitors (TKIs) such as imatinib, desatinib and nilotinib, bosatinib and ponatinib. In the diagnostic workup, qPCR is considered as the most sensitive and appropriate test for monitoring minimal residual disease (MRD) to define specific therapeutic milestones. Molecular monitoring data were analyzed in relation to TKI response for all the available cases of AP-CML. Results: We describe here seven AP-CML patients of which 6 (~85%) were females (median age 60 years) and only 1 (~15%) male (33 year) for which we performed qPCR. The male patient presented with high SOKAL and HASFORD prognostic risk score, was started with desatinib as firstline and achieved complete cytogenetic remission (negative by FISH) within 3-months of treatment with < 1% levels of p210 by qPCR. He is currently in major molecular remission (MMR, qPCR levels < 0.1% IS). No detectable comorbidities are reported till date. 2/6 (33%) females who received desatinib as first-line also achieved early cytogenetic and molecular response (3-months), and are currently in complete molecular remission (CMR, < LOD at 0.0003% IS). 4/6 (~67%) females who received either imatinib (n = 2) or desatinib (n = 2) as firstline were intolerant or refractory to treatment. Nilotinib (n = 3) and desatinib (n = 1, imatinib refractory case) treatments were able to achieve CMR (n = 3) and MMR (n = 1) in all the patients. The associated comorbidities or toxicities with TKIs observed were mainly hypothyroidism, type II diabetes, vitamin D deficiency, pleural effusion, hemorrhoids, hypercholesterolemia RV dilation and iron deficiency. Conclusion: In summary, depending upon the individual comorbidities AP-CML patients show a differential response to TKIs. In terms of better molecular outcome, desatinib and nilotinib show superior efficacy in APCML patients analyzed in this study. Disclosure: No conflict of interest disclosed. P514

Thrombocytopenia with hemophagocytosis: genetic clarification of a chromosomal translocation t(2;15) as potential underlying alteration Busemann C.1, Bakchoul T.2, Krüger W.1, Przybylski G.1, Pink D.3, Greinacher A.4, Schmidt C.A.2 Universitätsmedizin Greifswald, Hämatologie / Onkologie, Greifswald, Germany, 2Universitätsmedizin Greifswald, Greifswald, Germany, 3HeliosKlinikum Bad Saarow, Hämatologie / Onkologie, Bad Saarow, Germany, 4 Universitätsmedizin Greifswald, Transfusionsmedizin, Greifswald, Germany 1

Background: the pathogenesis of thrombocytopenia is not always clearly evident. Immunthrombocytopenia may be a diagnosis of exclusion, however, in many cases autoreactive antibodies are not detectable. Here we describe an unusual case of a young patient with accidentally diagnosed mild thrombocytopenia. Case: In a 25 year old male a thrombocytopenia of 50 GPT/l was diagnosed in the context of the evaluation of a gastrointestinal bleeding. The history of the patient was blank so far, no abnormal bleeding history was evident. Physical examination did not show any sign of diseases, especially no hematoma, lymph node or spleen enlargement. Routine blood checks including microscopic examination of a blood smear, routine blood chemistry as well as tests looking for idiopathic thrombocytopenia were uninformative and in the normal range too. Bone marrow examination revealed a prominent megakaryocytosis with mild dysplasia and a remarkable number of foaming histiocytes. Interestingly, these celles showed intracellular thrombocytes in various states of digestion. To identify a potential genetic background of this unusual finding, a cytogenetic analysis was performed and showed a constitutional translocation t(2;15) (q36–37;q21–22). To further delineate involvement of specific genes, next generation sequencing was performed on DNA extracted from peripheral blood neutrophils. The specific breakpoint was located between two genes

Abstracts

on both chromosomes each in noncoding regions without any known function. The precise breakpoint was confirmed with a specific PCR with primers flanking the putative breakpoint side on the original sample. Functional tests were undertaken to clarify whether abnormal thrombocyte behavior or false activation of the phagocytes are the driving force. Conclusion: we here describe the unusual case of a young patient with a constitutive chromosomal translocation and abnormal platelet phagocytosis. The clarification of the underlying pathomechanism might be of use to better understand aspects of hemophagocytosis. Disclosure: No conflict of interest disclosed. P515

Platelet volume is influenced by the applied aspheresis procedure Wenzel F.1, Bramhoff A.2, Giers G.2, Fischer J.2, Blessing F.3 Hochschule Furtwangen, Medical and Life Sciences, Villingen Schwenningen, Germany, 2Universitätsklinik Düsseldorf, Düsseldorf, Germany, 3Institut für Labormedizin, Singen, Germany 1

Introduction: Referring to current standards the quality of an apheresis procedure is estimated by the quantity of collected cells. Nowadays a new kind of quality measurement could be found in the detection of cell volumina. Recent diagnostics have shown that stem cells and platelets – when separated – are likely to appear in a higher volume inside the cell product. Therefore, in this study the question should be discussed wether platelets of higher volume are more likely to be separated than platelets showing a lesser volume. Methods: Blood samples of three different apheresis procedures could be observed: allogenic platelet donations (n = 7) (Trima, Terumo), autologous (n = 5) and allogenic stem cell donations (n = 5) (Cobe Spectra, Terumo). To examine the blood samples the Sysmex hematology analyser (XT-2000) has been used. Results: During stem cell apheresis, the volume of the separated platelets was 1.2fold increased compared to the platelet volume in the peripheral blood before separation. Before apheresis the mean platelet volume in the peripheral blood was found to be 6,21 fl, after apheresis 6,09 fl and inside the platelet concentrate 7,42 fl. The platelet number in the peripheral blood was also significantly decreased (before separation 180.1/nl and after separation 133.5/nl). In the blood products the concentration of platelets was nearly 8fold higher than in the peripheral blood before separation. Conclusion: Overall, the observed apheresis procedures are more likely to separate platelets showing a higher voulme than common in the peripheral blood. This might indicate that not only the amount of separated cells reflects the quality of the apheresis procedure but also that the volume of the separated cells can be used as a parameter for quality assessment. Disclosure: No conflict of interest disclosed. P516

Assessment of the T cell receptor repertoire in long-term platelet donors by next generation sequencing Link C.S.1,2, Hölig K.3, Rücker-Braun E.1, Lang K.4, Kuhn M.5, Eugster A.2, Klesse C.6, Schmiedgen M.1, Heidenreich F.1, Bornhäuser M.1,2, Bonifacio E.2, Schetelig J.1,6 Medizinische Klinik und Poliklinik 1, Medizinische Fakultät Carl Gustav Carus, TU Dresden, Hämatologie/Onkologie, Dresden, Germany, 2DFG Research Center for Regenerative Therapies Dresden, Dresden, Germany, 3 Medizinische Klinik und Poliklinik I, Universitätsklinikum Carl Gustav Carus, Fachbereich Transfusionsmedizin, Dresden, Germany, 4DKMS Life Science Lab, Dresden, Germany, 5Institut für medizinische Informatik und Biometrie (IMB), Medizinische Fakultät der TU Dresden, Dresden, Germany, 6DKMS Clinical Trials Unit, Dresden, Germany 1

Introduction: Platelet transfusions are needed in various clinical settings and have become a key part in the treatment of hematological diseases.

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When obtaining platelet products donors´ safety is of first priority. Multiple platelet donations have been reported to decrease lymphocyte count and this correlates with the number of donations. While this lymphopenia does not appear to be immunologically relevant from clinical experience, the immunological repertoire of platelet donors has not been measured directly. It remains unknown if the T cell receptor (TCR) repertoire is restricted after multiple platelet donations. Methods: Using next generation sequencing (NGS) we analyzed the TCRα repertoire of naive CD8+ T cells which had not been involved in an immune response and antigen experienced memory CD8+ T cells from five long-term platelet donors and five healthy controls. Diversity was estimated using Simpson’s diversity index (Ds). Results: A mean of 5,942,248 and 6,089,385 TCRα reads were found resulting in 60,082 and 12,058 clonotypes (mean) for the naive and memory compartment, respectively. Within all 10 samples we found a mean of 3,684 shared clonotypes (range 1,289 to 5,345) that were present in the naive and the memory repertoire, representing 11.35% (range 5.86% to 15.06%) of all TCRα reads of the naive cells and 75.13% (range 62.35% to 88.48%) of the reads in memory cells. Within the naive compartment none of the clonotypes was found with read frequencies >1% of reads. Diversity of the naive TCRα repertoire was 665fold higher than the diversity of the memory repertoire (p = 0.002). While the lymphocyte count was slightly lower for platelet donors compared to control samples (1.440 ± 0.178 *106/ml vs. 1.834 ± 0.174 *106/ ml, respectively) there were no significant differences in T cell subsets. Furthermore, comparison of the TCRα repertoires of long-term platelet donors and controls showed no differences in TCR diversity for the naive and memory compartment. Conclusion: To our knowledge this is the first study assessing the TCRα-repertoire in a healthy population by NGS. This dataset may help to define future reference criteria. The TCR-repertoire of regular platelet donors was not restricted compared to the repertoire of healthy control subjects. This finding supports the clinical judgement that lymphopenia in regular platelet donor does not implicate restricted immunity Disclosure: No conflict of interest disclosed. P517

Lack of sustained response to ponatinib treatment in myeloproliferative neoplasms associated with FGFR1 fusion genes Kreil S.1, Adès L.2, Bommer M.3, Stegelmann F.3, Ethell M.E.4, Lubking A.5, Martin P.6, Eigendorff E.7, Hofmann W.-K.1, Cross N.C.P.8, Hochhaus A.7, Reiter A.1 Universitätsmedizin Mannheim, III. Medizinische Klinik, Mannheim, Germany, Université de Paris, Hôpital Saint-Louis, Paris, France, 3Universitätsklinikum Ulm, Klinik für Innere Medizin III, Ulm, Germany, 4The Royal Marsden NHS Foundation Trust, Haemato-Oncology Unit, London, United Kingdom, 5Skåne University Hospital, Department of Hematology and Vascular Disorders, Lund, Sweden, 6MVZ Tempelhof, Berlin, Germany, 7Universitätsklinikum Jena, Klinik für Innere Medizin II, Jena, Germany, 8University of Southampton, Wessex Regional Genetics Laboratory, Salisbury, United Kingdom 1 2

Introduction: In patients with FGFR1 fusion gene positive MLN-eo (FGFR1+ MLN-eo), the optimal treatment remains to be defined. Encouraging in vitro data for inhibitors of the FGFR1 tyrosine kinase prompted the implementation of ponatinib into therapeutical strategies. In a cell free kinase assay, IC50 of ponatinib for FGFR1 was determined at 2.2 nM. Methods: In a retrospective survey, we sought to evaluate the efficacy of ponatinib in seven FGFR1+ MLN-eo patients. Results: Median age was 52 years (48–74) with a male predominance (n = 5). Cytogenetic analysis revealed a reciprocal translocation with involvement of chromosome band 8p11 in all patients [t(8;13)(p11;q12), n = 3; t(8;22)(p11;q11), n = 2; t(1;8;22)(?;p11;q11), n = 1; t(6;8) (q27;p11), n = 1]. RT-PCR identified the associated fusion genes ZMYM2-FGFR1 (n = 3), BCR-FGFR1 (n = 3), and FGFR1OP-FGFR1 (n = 1), respectively. All patients presented with left-shifted leukocytosis but only 3 [all with t(8;13)] had eosinophilia of >0.5 x 109/l. Five patients presented with con-

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comitantly diagnosed lymphoid neoplasms, i.e. T-lymphoblastic lymphoma (T-LBL, n = 3), biclonal accelerated phase (n = 1) or lymphoid blast phase of MPN/B-cell acute lymphoblastic leukemia (B-ALL, n = 1). All patients were initially treated with chemotherapy including hydroxyurea (n = 4) and/or high-dose regimens (n = 3), the latter exclusively in patients with concomitant lymphoid neoplasms. Lack of response, relapse or progression led to the use of ponatinib at a dose of 30mg/d (n = 2) or 45mg/d (n = 5), respectively. Median duration of treatment was 2 months (0.5–12), median observation time was 21 months (9–38). A temporary partial hematologic response was observed in 6 of 7 patients. One patient did not respond and died while on ponatinib for 2 weeks. Cytogenetic analysis was performed in 3 of 6 responders after median 3 months from start of ponatinib. Only one patient with t(8;13) achieved a partial cytogenetic response. Five patients underwent subsequent allogeneic stem cell transplantation (ASCT). They are in complete molecular remission and alive in median 22 months (9–38) after diagnosis and 15 months (5–31) after ASCT. One patient is on supportive care. Conclusions: Response to standard dose ponatinib in FGFR1+ MLN-eo has been poor. There was either progressive disease or no evidence for sustained complete hematologic or cytogenetic response. ASCT remains the only option to achieve long-term remission and possibly cure in FGFR1+ MLN-eo. Disclosure: No conflict of interest disclosed. P518

Treosulfan in combination with fludarabine as conditioning for allogeneic stem cell transplantation in myelofibrosis Kragl B.1, Heinz F.1, Große-Thie C.1, Henze L.1, Lakner J.1, Freitag S.1, Wittke C.1, Brueckner F.1, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany 1

Introduction: Allogeneic stem cell transplant (SCT) is a curative treatment option in selected patients with myelofibrosis. However, as the disease is rare little is known about best conditioning for SCT. Here, we present single center data on a treosulfan (Treo)/ fludarabine (Flud) based regimen. Method: Data of 1461 autologous and allogeneic transplants in our center since 1998 were analysed and we extracted 14 transplants (0.96%) with the indication of myelofibrosis (MF). Analysed data included: patient characteristics (age, sex, kind of MF), time first diagnosis (fd) to SCT, DIPSSscore at SCT, spleen size, conditioning regimen, mortality and cause of death. Results: One autologous SCT, 13 allogeneic SCT (all full HLA matched, 3 related, 10 unrelated). Graft source was bone marrow (n = 1) and peripheral blood stem cells (n = 13). Patient characteristics: median age 60 years (range 36–70 years), 10 male, 4 female patients; type of MF: 7 primary, 2 post ET-MF, 5 post PV-MF. Median time fd to SCT was 14.5 months (3–186 mths); DIPSS-score: high risk (n = 1), intermediate 2 (n = 6), intermediate 1 (n = 2), low risk (n = 5). Spleen was enlarged in 12 out of 14 patients. Conditioning regimen was Treo based in all cases (total dose: 36g/m² (n = 4), 42g/m² (n = 9), 50g/m² (n = 1)). Flud total dose was in general 150/m² divided onto 5 days. GVHD prophylaxis was cyclosporine A/ MTX based. Unrelated alloSCT recipients received in addition antithymoglobulin. GVHD occurred in 9 (70%) of 13 allogeneic tx. 30d mortality was 7% (n = 1), 60d mortality 21% (n = 3), 100d mortality 21% (n = 3) and 365d mortality 43% (6%). Median survival was 606d (range 12–3494). Cause of death were as follows: 6 transplant related (2 gvhd and sepsis, 3 sepsis, 1 sepsis and PTLD), 1 transformation in AML (autoSCT recipient, death +d1274). In this small cohort conditioning regimen, type of donor, spleen size and DIPSS-score had no impact on survival. It seems, that younger patient had a better outcome: 6 out of 7 living patients were ≤ 60 years old, but 4 out of 7 dead patients were >60 years old.

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Conclusions: SCT is increasingly applied to MF patients. Though a small cohort our data indicate that reduced intensity Treo/Flud conditioning is feasible. Infections following SCT are common causes of death. Disclosure: No conflict of interest disclosed. P519

Specific treatment significantly influences outcome in 32 pregnancies in polycythemia vera (PV) patients Wille K.1, Sadjadian P.1, Horstmann A.1, Kolatzki V.1, Griesshammer M.1 Johannes Wesling Klinikum Minden, Universitätsklinik für Hämatologie und Onkologie, RUB Bochum, Minden, Germany 1

Very limited data are available regarding pregnancy (preg) outcomes in patients with PV, less than 40 pregs being reported in the whole literature. Within a European Leukemia Net project we collected 32 pregs in 14 PV patients in our center since 2006. Pregs were categorized in two groups. Group 1 consisted of pregs receiving treatment according to a suggested algorithm (M. Griesshammer et al., Blood Rev. 2008) (n = 18). Group 2 (n = 14) included all pregs without a specific treatment. Most patients in group 1 received low dose aspirin during preg and low molecular weight heparin after delivery until the 6th week postpartum. The target hematocrit during preg was 40% and therapeutic phlebotomy was performed, if needed. Iron supplementation was not advised during preg. Median age at diagnosis of PV was 28.5 yrs (range 19–34), median age at delivery 33 yrs (24–39). Four out of 14 patients (28.6%) had high-risk PV due to severe thromboembolic complications either at the time of diagnosis or during follow up. These included 2 Budd Chiari syndromes, 1 portal vein thrombosis and 1 pulmonary embolism. Overall live birth rate was 47% (15/32). In group 1 live birth was recorded in 13/18 pregs (72%), while 5 pregs were unsuccessful due to spontaneous abortion. In contrast, outcome was significantly worse in group 2 with 2 live births out of 14 pregs (14%) [chi square, p = 0,001]. In group 2 we observed 8 (67%) spontaneous abortions, and 4 (33%) stillbirths. Interferon alpha was used in 4 cases with 3 live births (75%) and one spontaneous abortion. Concerning maternal complications, no thromboembolic events were observed. However, 5 bleedings occurred (4 minor and 1 major), all of them in group 1 which was probably due to the treatment with aspirin and/or low molecular weight heparin. Most PV patients’ hematocrit values spontaneously decreased during preg, thus phlebotomies were mainly performed during the first trimester. This is the largest series of PV pregs reported in a single center experience so far. The success rate of pregs was significantly better (72% versus 14%, respectively) for patients with appropriate management performed according to current guidelines. No thrombotic events were seen, however, an increased bleeding rate was observed.

with pt and disease characteristics, cytogenetics (CG) and molecular genetics (MG). Methods: Expression of CD30 was analyzed in BM samples by MFC using antibodies against CD30, CD45, CD117, CD2 and CD25. MC were identified as CD45 positive with bright CD117 expression. Coexpression of CD2 and/or CD25 defined neoplastic MC, median fluorescence intensities (medFI) of CD30 were determined and related to CD30 medFI in lymphocytes to derive CD30 index. MC infiltration and CD30exp by IH was assessed in 22 pt. CG and MG for KITD816V mutation were done in 44 and 80 pt, respectively. CD30exp detected by MFC was correlated to IH-detected CD30exp and to CG and MG. Results: Pt were 51 female, 42 male, median age 59 years (20–87), karyotypes: 41 normal, 3 aberrant, KITD816V mutation was positive in 74/80 pt. 14/93 pt had concurrent hematological non-mast cell disease (AHNMD). Mean (±SD) MC infiltration was 20%±26% (range, 1.5%-85%) by IH and 0.4%±1.8% (range, 0.01%-17%) by MFC. Mean (±SD) CD30 index was 19 ± 20 (range, 3–154), mean (±SD) CD30exp by IH was 9%±17% (range, 0%-70%). MC infiltration by IH and MFC correlated significantly (p = 0.002, r = 0.819), also KITD816V mutation ratio and MC infiltration by IH and MFC (p = 0.006, r = 0.669 and p < 0.001, r = 0.554, respectively). No correlation of MFC CD30 index with age, sex, AHNMD, KITD816V mutation ratio, grade of MC infiltration or %CD30+ MC by IH was found. Pt with normal vs aberrant karyotype had higher CD30 index (12.2 ± 6.2 vs 6.3 ± 2.7, p = 0.037). A trend to higher CD30 index in pt with KITD816V mutation was seen (23.9 ± 89.1 vs 10.0 ± 5.8, n.s.). Conclusions: Neoplastic MC harbouring aberrant karyotypes displayed stronger CD30 expression, being by trend also stronger on MC from pt with KITD816V mutations. CD30 expression on MC should be analyzed by both IH and MFC in a high number of pt to substantiate our findings and to better enable correlation with clinical outcome. Disclosure: Frauke Bellos: Employment or Leadership Position: angestellt im Münchener Leukämielabor Wolfgang Kern: Employment or Leadership Position: Teilhaber Münchner Leukämielabor P521

Two different coagulation factor VIII gene mutations in brothers with hemophilia A: an unexpected and rare finding Krammer-Steiner B.1, Nimtz-Talaska A.2, Toenges R.3, Friday D.3, Steiner M.4 Klinikum Südstadt Rostock, Klinik für Innere Medizin III, Rostock, Germany, Praxis für Kinder- und Jugendmedizin, Frankfurt/O., Germany, 3Diagenom GmbH, Rostock, Germany, 4Medizinisches Labor, Rostock, Germany 1 2

Introduction: Systemic mastocytosis (SM) is a rare disease with variable clinics, from indolent to aggressive clinical courses. Immunhistochemical staining (IH) has detected CD30 expression (CD30exp) on neoplastic mast cells (MC) as potential indicator for aggressive disease. CD30exp detected by multiparameter flow cytometry (MFC) might improve diagnostics and help correlate CD30exp with clinical parameters giving insights into disease biology. We compared CD30exp detected by MFC vs IH on bone marrow (BM) MC of patients (pt) with SM and correlated results

Introduction and Objectives: Inherited hemophilia A in females is rare and female hemophilia A patients are at risk of being misdiagnosed as acquired hemophilia A or as von Willebrand disease type 2N. Here we report a female patient with moderate factor VIII deficiency due to compound heterozygosity for two different coagulation factor VIII gene mutations. Material and methods: A 41-year-old mother presented her three sons aged 5, 8, and 12 years with moderately decreased factor VIII activity (20 to 24%) for genetic counseling and testing. Genomic DNA was obtained from anticoagulated blood. All factor VIII gene exons and flanking regions were amplified and PCR products were purified and sequenced. Results: Initial mutational screening identified the previously identified hemizygous factor VIII gene mutation p.Met2183Val (p.Met2164Val) located in exon 23 in the 12-year-old first son. Unexpectedly, the mutation could not be demonstrated in his two brothers prompting further full sequencing studies. A hemizygous mutation p.Arg2016Gln (p.Arg1997Gln) in exon 19 was identified in the 8-year-old and in the 5-year-old boys. Investigation of both mutations was conducted in the mother and confirmed the rare diagnosis of a compound heterozygous female hemophilia A patient. Her residual factor VIII activity was similar to that of her sons (23%) leading to the diagnosis of mild hemophilia A. Conclusions: Female cases of hemophilia A are rare. Among the potential genetic mechanisms leading to hemophilia A in females, compound hete-

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–160

Disclosure: No conflict of interest disclosed. P520

Assessment of CD30 expression on mast cells in systemic mastocytosis by immunohistochemistry versus multiparameter flow cytometry and correlation to clinical parameters Bellos F.1, Sotlar K.2, Jeromin S.1, Haferlach C.1, Haferlach T.1, Kern W.1 MLL Münchner Leukämielabor GmbH, München, Germany, 2Pathologisches Institut, Ludwig-Maximilians-Universität München, München, Germany 1

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rozygosity affecting both factor VIII alleles has been demonstrated only in a few cases. Here we report a female patient mother to three hemophilia A sons displaying two different missense mutations in exon 19 and exon 23, respectively. Compound heterozygosity was confirmed in the mother causing mild hemophilia A. Disclosure: No conflict of interest disclosed. P522

Haemophilia care: Which outcomes should be determined in clinical routine care and are adequate for access requirements Berger K.1, Eichler H.2, Escuriola-Ettinghausen C.3, Holstein K.4, Klamroth R.5, Königs C.6, Kurnik K.7, Oldenburg J.8, Scholz U.9, Schramm W.10, Tiede A.11 Klinikum der Universität München – Campus Grosshadern, Medizinische Klinik und Poliklinik III, München, Germany, 2Universität und Universitätsklinikum des Saarlands, Institut für Klinische Hämostaseologie und Transfusionsmedizin, Homburg / Saar, Germany, 3HZRM Hämophilie Zentrum Rhein Main GmbH, Mörfelden-Walldorf, Germany, 4Universitätsklinikum Hamburg-Eppendorf, Gerinnungsambulanz und Hämophiliezentrum, Hamburg, Germany, 5Vivantes Klinikum im Friedrichshain, Zentrum für Hämophilie und Hämostaseologie, Berlin, Germany, 6Universitätsklinikum Frankfurt, Goethe-Universität Frankfurt, Klinik für Kinder- und Jugendmedizin, Schwerpunkt Hämatologie, Onkologie und Hämostaseologie, Frankfurt am Main, Germany, 7Klinikum der Universität München, Kinderklinik und Kinderpoliklinik im Dr. von Haunerschen Kinderspital, München, Germany, 8Universitätsklinikum Bonn, Institut für experimentelle Hämatologie und Transfusionsmedizin, Bonn, Germany, 9 Zentrum für Blutgerinnungsstörungen, MVZ Labor Dr. Reising-Ackermann und Kollegen, Leipzig, Germany, 10Ludwig-Maximilians Universität München, RudolfMarx-Stiftung, München, Germany, 11Medizinische Hochschule Hannover, Klinik für Hämatologie, Hämostaseologie, Onkologie und Stammzelltransplantation, Germany 1

Background: Stakeholders like the Institute of Quality and Efficiency in Health Care (IQWiG), the Gemeinsame Bundesausschuss (G-BA) and payers are increasingly demanding quality oriented patient care. Clinical and patient reported outcomes are in the centre of this context. Therefore the working group “Outcomes” of the GTH-Panel Haemophilia reached out to determine relevant outcome parameters in clinical praxis and research to meet and evaluate the best possible patient care and meet the payers requirements. This also included assessment clinical routine care and how they could be made assessable for future research initiatives. Methods: Literature and desk researches to identify evidence requirements from different stakeholders’ perspectives; Development of a pilot questionnaire to prioritize clinical and patient reported outcomes (unimportant to very important); Questioning of physicians treating haemophilia patients (children and adults). Results: 14 physicians gave feedback on the questionnaire. Focus of the first evaluation was on clinical outcomes. Highly prioritized clinical outcomes were target joints (≥4 bleeds in one joint during 6 months), inhibitor development (influenced by the duration of treatment and intensity, kind of clotting factor concentrates, mutations), mortality and bleeds. All interviewees scored the aforementioned first three clinical outcomes as very important, 12 physicians graded bleeds (frequency, localization, severity) as very important, 1 as important. Arthropathy determined by clinical joint status, Haemophilia Joint Health Score (HJHS), WFH Physical Examination Score (Gilbert Score) has been graded as important by 6 and as very important by 8 interviewees. Determination of arthropathy by radiological joint scores eg Petterson score, magnetic resonance imaging and ultrasound were scored of moderate importance, therapy associated infections as important (n = 2), very important (n = 10), missing (n = 1). Osteoporosis has been indicated as an unimportant clinical outcome by 9 interviewees, 1 moderate important, 2 important. Conclusions: In a next step the working group “outcomes” will prioritize the patient reported outcomes to be collected. The group’s next objective is to develop a concept how to improve longitudinal outcome data collection in Germany. Disclosure: No conflict of interest disclosed.

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P523

Platelet transfusion in oncology/hematology patients in routine clinical practice in of a tertiary hospital Berger K.1, Wittmann G.2, Henschler R.3, Rieger C.1, Ostermann H.1 Klinikum der Universität München – Campus Grosshadern, Medizinische Klinik und Poliklinik III, München, Germany, 2Klinikum der Universität München, Abteilung für Transfusionsmedizin, Zelltherapeutika und Hämostaseologie, München, Germany, 3Schweizerisches Rotes Kreuz SRK, Blutspende Zürich, Zürich, Switzerland 1

Introduction: There is little information on real life patient data prior and post transfusion of blood components of clinical hematology and oncology. The objective of this study is to describe patient demographics, indication of platelet transfusion, to compare actively collected hemovigilance data and to determine adherence to transfusion guidelines and relative effectiveness for patients transfused with platelet components in routine mostly prophylactic use. Methods: Prospective open label, single-center, observational study. Inclusion of platelet transfusions was consecutive. During the observation period the platelet concentrates (plt) transfused were either pathogen inactivated (PI) or produced conventionally (CON). Results: Between March and May 2015 126 patients (pts) received 1207 platelet units (586 PI plts, 621 CON plts). 105 patients (pts) signed consent, 11 pts were not able to give informed consent. Subsequently data for 464 PI plt and 482 CON plt could be documented. Mean age was 53.8 years. 39% pts were younger than 50 years. 63% of pts were male, 80% of pts were diagnosed with haematological diseases. In total 5 adverse events were documented: 1 AE following PI, mild severity not requiring further medical invervention,4 AEs following CON platelet transfusion of moderate severity which required further medical intervention. 10% of pts with underlying haematological diseases received more than 30 transfusions (36% of all transfused plt). In this group approx. 69% plts were transfused at a transfusion trigger > 10 G/I. 6% of pts received 20 -30 transfusions (14% of all transfused plt) 18% of pts received 10–20 transfusions (24% of all transfused plt), 66% of pts received less than 10 plt transfusions (26% of all transfused plt). Conclusions: The majority of all platelet concentrates in this study was transfused in a small number of patients. Only a limited number of transfusion reactions were documented. We found a trend to a reduced incidence of adverse events for the PI treated platelet concentrates. Platelets were mostly transfused according to current treatment guidelines. Disclosure: Karin Berger: Expert Testimony: Research Grant CERUs Helmut Ostermann: Expert Testimony: Research Grant CERUS

Posterdiskussion

Aggressive Non-Hodgkin-Lymphome P524

High efficiency of the pan-PI3K/mTOR inhibitor, PQR-309, in MCL Kvint R.1, Zimmermann Y.1, Hutter G.1, Hiddemann W.1, Dreyling M.1 Medizinische Klinik III, Klinikum der Universität München, München, Germany

Introduction: Mantle cell lymphoma (MCL) comprises about 6% of all non-Hodgkin´s lymphoma with a median survival of 3–5 years. New emerging strategies especially include inhibitors of the PI3K/mTOR signalling pathway (PI3K, Akt, mTOR) which is constitutively activated in MCL and plays a critical role in tumor growth and survival. In this study we evaluated the efficiency and mode of action of the dual PI3K/mTOR (c1+c2)-Inhibitor, PQR-309, in MCL cell lines, as a single agent but also in combination with others molecular Inhibitors. Methods: MCL cell lines (Granta 519, Jeko-1, Rec-1 and Mino) were exposed to the panPI3K/ double mTOR inhibitor (PQR-309) as well as to combination of PQR-309 with PIM- (AZD-199), BET- (JQ1), MEK(PD-0325901); BCL2- (ABT199); BTK-(Ibrutinib); RSK-(BI-D1870);

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STAT3-(Niclosamide)and JAK-Inhibitor (Ruxolitinib). Cell proliferation (trypan blue staining), apoptosis (Annexin V PE/7-AAD staining) and cell cycle (FACS) were investigated. Protein expression and phosphorylation status of downstream proteins (Akt, mTor, Raptor, Rictor, MAPK/ MEK, Rb, 4EBP1) were analysed at 1h, 4h, 8h and 24h of treatment. Results: PQR-309 appears to be an effective inhibitor in MCL. Treatment with PQR led to a reduction of cell proliferation in all MCL cell lines due to a G1 cell cycle arrest. There was no apoptosis detectable. Thereby the impact of PQR-309 on viable cell count was stronger than that of a single PI3K-inhibitor (CAL101), single mTOR-Inhibitor (Temsirolimus) and even that of the dual mTOR-Inhibitor, KU-0063794, targeting mTORC1 and mTORC2. In all cell lines dephosphorylating of mTor, Raptor, Rictor, Akt was detected after treatment with PQR, again with a less prominent effect under treatment with CAL101, Temsirolimus and KU separately. The decrease of phosphorylated proteins occurred during the first hour of treatment. Combination of PQR with PIM was most efficient und shows strong synergism. This strong synergism was accompanied by the dephosphorylation of a downstream target of the PI3K/AKT/mTOR-Pathway and the PIM1 phosphorylation target, 4EBP1. Conclusions: The combination of PQR-309 with the inhibitor of PIM kinases shows an impressive efficiency in MCL cell lines. Cotargeting PIM kinases, PI3K and mTOR represents a promising novel approach in MCL. Disclosure: Rossana Kvint: No conflict of interest disclosed. Martin Dreyling: Advisory Role: Janssen; Financing of Scientific Research: Janssen, Roche; Expert Testimony: Janssen, Roche P525

Pharmacological modulation of the NOXA-MCL1 balance is an effective strategy for treatment of mantle cell lymphoma Höring E.1, Montraveta A.2, Heine S.3, Kleih M.3, Schaaf L.3, Vöhringer M.1, Ott G.4, Campo E.2, Colomer D.2, Aulitzky W.E.1, van der Kuip H.3 Robert-Bosch-Krankenhaus, Hämatologie/Onkologie, Stuttgart, Germany, Center Esther Koplowitz, Institut d‘Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Hospital Clinic, Hematopathology, Barcelona, Spain, 3Dr. Margarete Fischer-Bosch-Institut für Klinische Pharmakologie, Stuttgart, Germany, 4Robert-Bosch-Krankenhaus, Klinische Pathologie, Stuttgart, Germany 1 2

Mantle cell lymphoma (MCL) is a rare lymphoid neoplasm often characterized by an aggressive clinical course with early relapses. The interplay between pro- and anti-apoptotic proteins of the BCL-2-family is crucial in regulating mitochondrial apoptosis in hematopoietic cells and imbalances in the composition of its members contribute to tumorigenesis and therapy resistance. We recently demonstrated that MCL cells protect themselves against apoptosis by rapidly degrading NOXA protein. Therefore, pharmacological stabilization of NOXA is a promising strategy to treat MCL. Indeed, we could show that proteasome inhibitors (e.g. Bortezomib) and fatty acid synthase inhibitors (FASNi) (e.g. Orlistat) effectively induce cell death in MCL by accumulating NOXA protein. However, the efficiency of apoptosis induction at lower concentrations was limited despite stabilization of NOXA as concomitant induction of the anti-apoptotic MCL1 counteracts induction of cell death. We therefore investigated whether dual targeting of NOXA and MCL1 could be a promising strategy to enhance effectiveness of apoptosis induction in MCL. The CDK-9 inhibitor Dinaciclib has been described to potently inhibit MCL1 transcription. Here we show that this compound rapidly reduces MCL1 protein in a dose- and time-dependent manner in MCL cells. To study the efficiency of dual targeting of NOXA/MCL1 we combined sublethal doses of Dinaciclib and NOXA-stabilizing agents and observed synergistic induction of apoptosis in MCL cell lines (Mino, JeKo1, Jvm2). Combined treatment significantly accumulated NOXA and abrogated MCL1 induction. Cell death was dependent on NOXA as siRNA-mediated suppression of NOXA almost completely abolished induction of apoptosis. Importantly, combined treatment of Dinaciclib with the FASNi Orlistat was also highly effective in primary cells from MCL patients and signifi-

Abstracts

cantly inhibited tumor progression in vivo in a MCL mouse model. Analogous to cell line models, we observed NOXA protein accumulation and concomitant MCL1 downregulation. Interestingly, we found a significant correlation between induction of cell death and the NOXA/MCL1 protein ratio highlighting the importance of modulating the NOXA-MCL1 axis for effective induction of cell death in MCL. In summary, the NOXA-MCL1 balance is critical for survival of MCL cells. Dual targeting of MCL1 and NOXA efficiently kills MCL in vitro and in vivo and appears to be a promising strategy to be tested in clinical trials. Disclosure: No conflict of interest disclosed. P526

Synergistic effect of inhibition of multiple PI3K-isoforms due to the differential mode of action of single PI3K-isoform inhibitors in mantle cell lymphoma Hutter G.1,2, Zimmermann Y.1,2, Bamopoulos S.A.1, Irger M.1,2, Hiddemann W.1,2, Dreyling M.1,2 Klinikum der Universität München – Campus Grosshadern, München, Germany, ELLF, München, Germany

1 2

Introduction: Mantle cell lymphoma (MCL) is a distinct lymphoma subtype with an aggressive clinical course and a median survival of 3–5 years. New emerging strategies include especially inhibitors of the B-cell receptor pathway which is constitutively activated in MCL and plays a critical role in tumor growth and survival. In the present study we investigated different PI3K-Inhibitors targeting different isoforms. Methods: MCL cell lines (Z-138, Mino-1, Granta-519, Jeko-1, Rec-1, Maver-1) were exposed to different PI3K-Inhibitors (BAY-80–6946, Pictilisib, AS-605240, IPI-145, A66, CAL-101 and TGX-221). The effect of drugs was evaluated by cell count (Vicell), cell metabolism (WST-assay), cell cycle (FACS) and apoptosis (Annexin V PE/7-AAD staining). Subsequently, combinations with other molecular inhibitors were analysed. Proteome Profiler Phospho-Kinase Antibody Array as well as Western blot analyses were performed after exposure to the various PI3K Inhibitors. Efficiency of drug combinations were confirmed in primary patient samples. Results: For the single PI3K isoform inhibitors AS-605240 was the most efficient reaching a median response rate of 26% (8,2%-49,7%). Comparison of single isoform inhibition to combined inhibition of the different isoforms in MCL cell lines confirmed the superiority of the combined inhibition of all four isoforms (overall response rate 50%). The differential impact of single and multiple PI3K isoform inhibition on the phosphorylation status of Phospho Kinases lets suggest that concurrent dephosphorylation of Akt on Ser473 and Thr308, RSK1/2/3, JNK, LCK, PRAS40, c-Jun, p27 and WNK1 is responsible for increased response rates to the inhibition of all four PI3K-isoforms. Thereby targeting different isoforms of PI3K revealed different proteinphosphorylation patterns leads suggest that the different PI3K isoforms act on different downstream targets. providing an explanation for the superiority of inhibition of multiple PI3Kisoforms. Deductive from the proteinphosphorylation pattern potential-effective combination inhibitors were tested together with PI3Kinhibitors in MCL cell lines. Conclusion: Targeting different isoforms of PI3K revealed different proteinphosphorylation patterns supporting the assumption that the different PI3K isoforms act on different downstream targets. This provides an explanation for the higher efficiency of inhibition of multiple isoforms of PI3K inhibitors. Disclosure: Grit Hutter: No conflict of interest disclosed. Martin Dreyling: Advisory Role: Janssen; Financing of Scientific Research: Janssen, Roche; Expert Testimony: Janssen, Roche (Institution)

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Combinations containing the aza-anthracenedione pixantrone (Pix) show pre-clinical activity in B-and T cell lymphoma Tarantelli C.1, Gaudio E.1, Kwee I.1, Stathis A.2, Zintl P.3, Zucca E.2, Bertoni F.1,2 IOR Institute of Oncology Research, Bellinzona, Switzerland, 2IOSI Oncology Southern Switzerland, Bellinzona, Switzerland, 3CTI Life Sciences, London, United Kingdom 1

Introduction: DLBCL is the commonest lymphoma and 1 quarter of patients still present with a refractory disease or relapse after first line chemotherapy. Pix is an aza-anthracenedione with reduced cardiotoxicity, which has received a conditional marketing approval in the E.U. as a monotherapy for the treatment of adults with multiply relapsed or refractory aggressive B-cell non-Hodgkin lymphomas. Here, we evaluated the pre-clinical activity Pix with a series of additional anti-lymphoma drugs in cell lines derived from DLBCL and T-cell lymphoma. Methods: DLBCL cell lines derived from activated B-cell like (ABC) DLBCL, from germinal center B-cell (GCB) DLBCL and T-cell lymphoma cell lines were studied. Pix was used in combination with: bendamustine, ibrutinib, idelalisib, lenalidomide, rituximab, vorinostat, bortezomib and etoposide. Due to the mechanism of action, lenalidomide, bortezomib and ibrutinib were evaluated only in ABC-DLBCL cell lines. Cell lines were exposed (72 h) to increasing doses of the compounds alone or in combination, followed by MTT assay. The Chou-Talalay combination index (CI) was estimated using the Synergy R package: CI < 0.9, synergism; CI between 0.9 1.1, an additive effect; CI >1.1, no benefit. Results: The median IC50 of Pix as single agent was 200 nM among 14 DLBCL cell lines. GCB-DLBCL presented a trend for a higher sensitivity to the drug than ABC-DLBCL (P = 0.05). The combination with the BTK-inhibitor ibrutinib was effective in 6/7 ABC-DLBCL (median CI = 0.67, range 0.33–1.44): synergism in 4 and additive effect in 2. The combination with the PI3K-delta inhibitor idelalisib was beneficial in 11 out of 12 DLBCL (median CI = 0.67; range 0.1–1.05): synergism in 10 and additive effect in 1. Benefit was also observed when Pix was combined with lenalidomide in 2/2 ABC-DLBCL (synergism and additive effect, 1 each), with the anti-CD20 monoclonal antibody rituximab in 2/4 DLBCL (synergism in both) and with the HDAC-inhibitor vorinostat in 2/4 DLBCL. A synergistic effect was seen when Pix was combined with etoposide in 3/4 DLBCL and in 2 out of 3 T-cell lymphoma cell lines. Conclusions: Pix was very active in DLBCL cell lines as single agent and benefit was observed when the drug was combined with different additional anti-cancer agents. The combinations with the BTK inhibitor ibrutinib and with the PI3K-delta inhibitor idelalisib were the most effective and are worth of further studies at pre-clinical and clinical level. Disclosure: Chiara Tarantelli: No conflict of interest disclosed. Francesco Bertoni: Expert Testimony: Von CTI P528

HCV load as a possible prognostic factor in patients with HCVrelated DLBCL Zhou X.1, Lisenko K.1, Lehners N.1, Brandt J.1, Cremer M.1, Kriegsmann M.2, Ho A.D.1, Witzens-Harig M.1 Heidelberg University, Department of Hematology, Oncology and Rheumatology, Heidelberg, Germany, 2Heidelberg University, Institute of Pathology, Heidelberg, Germany 1

Introduction: A causal relationship between hepatitis C virus (HCV) infection and diffuse large B-cell lymphoma (DLBCL) has been suggested in several studies. Rituximab, cyclophosphamide, hydroxydaunorubicin, vincristine, prednisone (R-CHOP) is considered as standard first line therapy for HCV-related DLBCL. In contrast, the role of HCV eradication in the management of HCV-related DLBCL is still under debate. The aim of this study was to analyze the clinical characteristics and outcome of

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patients diagnosed with both HCV infection and DLBCL and to identify possible prognostic factors, in particular focusing on HCV load. Methods: We performed a retrospective single center analysis of patients with DLBCL co-diagnosed with a HCV infection. A total of 15 patients were identified. Patients’ demographic characteristics, data on DLBCL and HCV diagnosis, treatment and outcome were evaluated. Kaplan-Meier survival analysis and for comparison of survival rates between groups a log-rank tests was performed (GraphPad Prism 5.0). Results: The majority of patients (n = 10, 67%) was diagnosed with DLBCL in an advanced stage. Three patients (20%) had secondary transformed DLBCL deriving from a follicular lymphoma. 67% of patients (n = 10) obtained CHOP-like chemotherapy. Two thirds of patients (n = 10, 67%) were diagnosed with HCV infection prior to and one patient post DLBCL diagnosis. In three cases HCV infection and DLBCL were diagnosed simultaneously. Genotype 1 was the most common subtype of HCV (n = 11, 73%). Four patients (27%) received ribavirin and pegylated interferon-α while the remaining patients did not receive any HCV-specific therapy. A HCV/Hepatitis B virus (HBV) co-infection was documented in six patients (40%). In total, five patients died in the course of the disease. Two patients had a rapid progression of DLBCL and deceased during first line chemotherapy. In three patients, the cause of death was unknown. Follow-up HCV load of ≥1×106 IU/ml and maximal HCV load of ≥1×107 IU/ml were associated with an inferior overall survival (P = 0.02). Conclusions: Our findings suggest that HCV load could represent a negative prognostic factor in HCV-related DLBCL. Therefore, HCV eradication should be considered in patients with DLBCL and detectable HCV load. However, due to the retrospective design and the small number of cases, further larger studies are needed to investigate our findings and the role of antiviral therapy in the management of HCV-related DLBCL. Disclosure: No conflict of interest disclosed. P529

Incidence of CNS relapse after treatment of DLBCL, greyzone lymphoma and Burkitt lymphoma with R-DA-EPOCH – a single-center analysis Leng C.1, Korfsmeier K.1, Pezzutto A.1 Universitätsmedizin Berlin/Charité, Hämatologie und Onkologie, Berlin, Germany 1

Introduction: R-DA-EPOCH has shown convincing activity in aggressive B cell lymphoma. We set out to validate the results in our medical center and to evaluate the incidence of CNS relapse. Methods: Institutional databases were queried identifying patients receiving R-DA-EPOCH as frontline therapy between 2010 and 2014. Diffuse large B cell lymphoma (DLBCL), including HIV- and EBV-associated B cell lymphoma, primary mediastinal B-cell lymphoma (PMBCL), grey zone lymphoma and Burkitt-lymphoma were included. Use of CNS prophylaxis was at the discretion of the treating physician. Risk for CNS relapse was defined as follows: at least two extranodal lesions, elevated LDH and/or bone marrow infiltration. Patient characteristics, outcome with therapy, time to therapy failure and CNS relapse were retrospectively analyzed. Results: 48 patients (pts) were identified: DLBCL in 35 pts (73%), PMBCL in 4 pts (8%), grey zone lymphoma in 7 pts (15%) and Burkitt lymphoma in 2 pts (4%). 1 patient had double hit lymphoma (translocations of c-myc and BCL6). 9 pts had HIV-associated B cell lymphoma, 6 pts EBV-associated B cell lymphoma. 8 pts (17%) had ECOG PS>1, 38 pts (79%) had LDH above upper limit of normal, 35 pts (73%) had stage III-IV disease and 18 pts (38%) had two or more EN sites of disease, including 2 pts with renal manifestation. 13 pts (27%) had IPI 0-1, 23 pts IPI 2–3 (48%) and 12 pts IPI 4–5 (25%). A total of 8 (17%) pts recieved CNS prophylaxis (i. th. In 6 pts, HD-MTX in 2 pts). 22 pts (48%) achieved complete remission, 16 pts (33%) achieved partial remission and 2 pts (4%) showed stable disease. 8 pts (17%) exhibited progression during therapy. 24-month PFS was 73%. 17 pts (34%) showed a risk profile for CNS relapse. A total of 3 pts (6%) had finally developed a CNS relapse (intraparenchymal in all three cases)

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with fatal outcome, occurring at a median of 7 months after start of therapy (range 6 -17 months): the only patient with double hit lymphoma, one of 2 pts with renal involvement and one without the defined risk factors (relapse occurring 12 month after primary therapy). None of the patients received systemic CNS prophylaxis. Conclusion: Our retrospective analysis confirms the effectivity of R-DA-EPOCH in aggressive B cell lymphomas. The risk of CNS progression in patients receiving R-DA-EPOCH was similar to previous reports with R-CHOP. Thus, it appears that additional systemic CNS-prophylaxis is warranted in high risk patients. Disclosure: No conflict of interest disclosed. P530

Subcutaneous rituximab in patients with DLBCL and follicular NHL – First interim results from the non-interventional study MabSCale Dürig J.1, Azeh I.2, Blumenstengel K.3, Gerhardt A.4, Hänel M.5, Hapke G.6, Hensel M.7, Heßling J.8, Höffkes H.-G.9, Hurtz H.-J.10, Ritter M.11, Staib P.12, Uhlig J.13, Marquardt M.14, Krumm K.14, Fischer von Weikersthal L.15 Universität Duisburg-Essen, Klinik für Hämatologie, Essen, Germany, OnkoLogiX GmbH & Co. KG, Gelsenkirchen, Germany, 3Onkologische Praxis, Eisenach, Germany, 4Medizinisches Versorgungszentrum für Blut- und Krebserkrankungen, Potsdam, Germany, 5Klinik für Innere Medizin III der Klinikum Chemnitz gGmbH, Chemnitz, Germany, 6Kath. Marienkrankenhaus Zentrum für Innere Medizin, Hamburg, Germany, 7Mannheimer Onkologie Praxis Dres. Brust-Schuster-Plöger-Prof. Hensel, Mannheim, Germany, 8 Onkologie am Segelfliegerdamm, Berlin, Germany, 9Medizinisches Versorgungszentrum Osthessen GmbH, Fulda, Germany, 10Onkomedic GbR, Halle, Germany, 11Klinikverbund Südwest -Kreiskliniken Böblingen gGmbH, Sindelfingen, Germany, 12St. Antonius-Hospital Eschweiler, Klinikum für Hämatologie und Onkologie, Eschweiler, Germany, 13Facharztpraxis für Innere Medizin Hämatologie und Onkologie, Naunhof, Germany, 14Roche Pharma AG, Grenzach-Wyhlen, Germany, 15Gesundheitszentrum St. Marien Amberg, Amberg, Germany 1 2

Introduction: Clinical studies demonstrated rituximab (R) to be effective in combination with standard chemotherapies (CT) in the treatment of follicular lymphoma (FL) and diffuse large B-cell lymphoma (DLBCL). Aim of this study is to evaluate the efficacy and safety of the subcutaneous administration. Methods: Adult patients (pts) with previously untreated CD-20 positive DLBCL or FL to receive R and CT as first line treatment will be enrolled. Treatment decisions are at physician’s discretion. An interim analysis was planned to receive early impressions about efficacy and safety after 400 pts (data cut 04/03/2016). Primary endpoint is the (unconfirmed) complete remission CR/CRu rate. Secondary objectives include 2-years-progression-free-survival (PFS) for FL pts with maintenance therapy, the safety profile in induction (all pts) and maintenance therapy (only FL pts) and the analysis of treatment modalities. Results: 377 pts were included in this interim analysis (171 FL; 206 DLBCL) of these 74.3% pts were still ongoing in the FL group, 30.6% in the DLBCL group. Baseline characteristics in the FL/DLBCL subsets were as follows: median age 66/69.5 years; elevated LDH 42.7%/56.8%; 23%/56% had early stage disease (Ann Arbor I/II). The FL international prognostic index (FLIPI) indicated low/ intermediate/ high risk for 20%/35%/46% FL pts. The IPI classification of DLBCL pts with low/low-intermediate/ high-intermediate/high risk was 38%/39%/18%/5%. The most frequently used CT combination partner was bendamustine in 69% of FL pts, whereas DLBCL pts were treated with CHOP-21 (43%) and CHOP-14 (40%). A sensitivity analysis regarding pts with completed induction or study discontinuation (FL: N = 124, DLBCL: N = 143) revealed CR/CRu rates of 21.8% (95%CI: 15.4; 29.8) and 32.2% (95%CI: 25.1; 40.2) with an ORR of 60% (FL) and 63% (DLBCL) The dropout rate was about 30% in both groups, where study discontinuations were mostly due to patient’s decisions and rarely caused by AE. In the FL/DLBCL cohort AEs of any grade and of ≥ grade 3 were reported for 77.2%/76.7 and 33.3%/38.8%

Abstracts

respectively. For 24%/29.1% of pts serious AEs of any grade were documented, with 18.1%/25.2% classified as ≥ grade 3. Conclusions: Results from this preliminary analysis are limited by the low number of pts having completed induction treatment and thus need to be interpreted with caution. More mature data can be expected at a later stage in time and will be presented at the conference in autumn 2016. Disclosure: Jan Dürig: Advisory Role: Teilnahme an Advisory Boards der Fa Roche; Financing of Scientific Research: Honorare für Vortragstätigkeit von der Fa Roche Ludwig Fischer von Weikersthal: Other Financial Relationships: Congress Travel Expenses Fa Roche P531

Evaluation of safety, tolerability and efficacy of Temsirolimus in patients (pts) with relapsed or refractory mantle cell lymphoma (rel/refr MCL) in routine clinical practice Dreyling M.1, Krekeler G.2, Neuhof A.2, Woike M.2, Hess G.3, Kalanovic D.2 Klinikum der Universität München – Campus Grosshadern, Medizinische Klinik III, München, Germany, 2Pfizer Pharma GmbH, Berlin, Germany, 3 Universitätsmedizin der Johannes Gutenberg-Universität, III. Medizinische Klinik, Mainz, Germany 1

Background: Temsirolimus (TEM), an mTOR inhibitor, is approved in the EU for the treatment of pts with rel/refr MCL. A pivotal study demonstrated significantly longer progression free survival (PFS) with TEM (175 mg weekly for 3 weeks followed by 75 mg weekly) in rel/refr MCL pts compared to investigator´s choice therapy (4.8 vs 1.9 months (mo); p = .0009). Yet only limited data is available on TEM in an unselected routine clinical patient population. To evaluate the safety profile and efficacy of TEM in this rare tumor entity, further collection of data in a post-approval prospective non-interventional trial is useful. Methods: A German multicenter registry for rel/refr MCL pts treated with TEM was started in Germany in Oct 2009 (NCT00700258) with regulatory and ethic committee´s approval. Objectives are the evaluation of the safety profile, tolerability and anti-tumor activity of TEM as well as patient’s profile including comorbidities, characteristics, and the sequence of systemic therapies. Results: From Oct 2009 to Feb 2016, 53 pts were recruited in 28 study sites. Baseline characteristics are available for 53 pts: 69.8% male; median age 74.4 years; bone marrow involvement in 39.6% of the pts; ECOG performance status (n = 52) 0 or 1 in 82.7%, ECOG PS 2 in 17.3%. According to MIPI score (n = 51), 21.6%, 33.3%, and 45.1% are classified as low, intermediate, and high risk at the time of enrollment. The median number of prior therapies is 2 (range 1–10) with 43.4% treated in ≥ 4th line. Most common adverse events (in ≥ 20% of the pts) are thrombopenia (47.2%) and anemia (22.6%). Severe adverse events (drug related) are general, metabolic, psychiatric, skin, renal, gastrointestinal, respiratory (in 1 pt each) and blood system disorders (in 2 pts) and infections (in 4 pts). Efficacy analyses are available for 38 assessable pts and show an objective response in 31.6%, a clinical benefit (CR, PR, MR and SD) in 60.5% and PD in 39.5% of the pts. Median PFS for all pts is 3.7 mo. For the subgroup of pts treated with TEM in 2nd and 3rd line PFS is 3.3 mo, for ≥ 4th line pts 4.9 mo. Conclusions: The registry was started to evaluate the safety and efficacy of TEM in pts with rel/refr MCL in routine clinical practice. 45.1% high-risk pts were included in the analysis. In this comparatively poor-prognosis patient population, TEM showed a predictable, manageable tolerability profile. Efficacy parameters were consistent with published phase III data. Disclosure: Martin Dreyling: Advisory Role: Pfizer Consultant and Advisory Role; Financing of Scientific Research: Pfizer Daniel Kalanovic: Employment or Leadership Position: Employment and Leadership Position

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Efficient stem cell collection after modified cisplatin-based mobilization chemotherapy in patients with DLBCL Lisenko K.1, Cremer M.1, Schwarzbich M.-A.1, Kriegsmann M.1, Ho A.D.1, Wuchter P.1, Witzens-Harig M.1 University Heidelberg, Department of Hematology, Oncology and Rheumatology, Heidelberg, Germany 1

Introduction: R-DHAP (rituximab, dexamethasone, cytarabine, cisplatin) is an effective salvage therapy regimen in patients with relapsed or refractory diffuse large B-cell lymphoma (DLBCL) and is also used for mobilization of peripheral blood stem cells (PBSC). At our center a modified regimen with administration of 25 mg/m2 cisplatin as a 3-hour infusion over 4 consecutive days instead of the usual standard dose of a single infusion of 100 mg/m2 over 24 hours has been established to minimize renal toxicity. We performed a systematic evaluation of this modified R-DHAP salvage regimen for the mobilization and collection of PBSCs. Methods: We analyzed retrospectively clinical characteristics, PBSC collection and autologous stem cell transplantation (ASCT) parameters, as well as hematological reconstitution data of 65 patients with relapsed or refractory DLBCL who underwent PBSC collection after mobilization with the modified R-DHAP protocol at our institution between 2002 and 2013. Data were evaluated for the overall cohort and with regard to the number of R-DHAP cycles received prior to PBSC collection. Results: PBSC collection was performed after the first, second or third R-DHAP course in 32 (49%), 30 (46%) and 3 (5%) patients, respectively. 63 (97%) patients reached the collection goal of ≥2.0×106 CD34+ cells/kg body weight (bw). A significantly higher median CD34+ collection yield was reached when cells were collected after the first compared to the second R-DHAP course (p < 0.01). A peripheral blood leukocyte increase of ≥1.0 x109/l and platelets increase ≥20 x109/l was observed 11 days after ASCT. Conclusion: Our data show that a modified R-DHAP regimen with prolonged administration of cisplatin 25 mg/m2 over 4 consecutive days is an effective and safe treatment option that allows successful PBSC mobilization in relapsed or refractory DLBCL. Disclosure: Katharina Lisenko: No conflict of interest disclosed. Mathias Witzens-Harig: Advisory Role: Celgene; Financing of Scientific Research: Roche P533

Reliability of CT-based detection of bone marrow involvement in DLBCL Pecher A.-C.1, Soekler M.1, Kopp H.-G.1, Kanz L.1, Horger M.2, Müller M.1 Medizinische Klinik und Poliklinik II, Abteilung für Onkologie, Hämatologie, Rheumatologie, Klinische Immunologie und Pulmonologie, Tübingen, Germany, 2 Radiologische Universitätsklinik, Tübingen, Germany 1

Bone marrow (BM) involvement is an important prognostic marker in Non-Hodgkin lymphoma (NHL). Knowledge of the patient’s BM status is required for Ann-Arbor staging and adequate International prognostic Index (IPI) calculation. A histopathological evaluation of a BM biopsy is currently viewed as the gold standard to rule out BM involvement. The reliability of CT scan as a tool to rule out BM involvement is not known. Here, we retrospectively analyzed the performance of CT scan to predict BM involvement in n = 141 patients with firstly diagnosed diffuse large B cell lymphoma (DLBCL), who were diagnosed at our institution from 2013–2015. 15% of patients with DLBCL (n= 21) showed BM involvement at diagnosis as assessed by histopathological examination. 19% of the cases with BM involvement (n = 4) were not detected by CT-scan. In n = 6 patients (4.2%) BM involvement was suggested by CT scan but could not be confirmed by BM biopsy. Thus, CT-scan showed a sensitivity of 81% and a specificity of 95% for the prediction of BM involvement in DLBCL. The positive predictive value (PPV) was 73.9%, while the negative predictive value (NPV) amounted to 96.6%.

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In summary, our analysis demonstrates that CT scan is a diagnostic tool to rule out bone marrow involvement with a very good NPV of 96.6%. Due to a significantly lower PPV of 73.9%, suspected BM involvement on CT scan should generally prompt a BM biopsy to rule out false positives. Taken together, BM biopsy remains the gold standard to rule out BM involvement in DLBCL but CT scan can be applied in selected cases where patients are not able or refuse to undergo this procedure. Disclosure: No conflict of interest disclosed. P534

Ibrutinib-induced rapid response in chemotherapy-refractory Richter`s Syndrome Fischer-Maranta A.1, Bastian S.1, Mey U.1, Cogliatti S.2, Hohloch K.1,3 Kantonspital Graubünden, Hämatologie und Onkologie, Chur, Switzerland, Kantonspital St. Gallen, Institut für Pathologie, St. Gallen, Switzerland, 3 Uniklinik Goettingen, Hämatologie/Onkologie, Goettingen, Germany 1 2

Introduction: Richter`s syndrome describes the transformation of CLL into aggressive lymphoma, mostly into DLBCL. Prognosis is poor with a response rate of <20% after anthracycline-based chemotherapy and with a median survival of <12 months after transformation. Stem cell transplantation (SCT) improves survival, however, only 15% of patients ultimately undergo SCT due to primary-refractory disease. Ibrutinib, a bruton tyrosine kinase inhibitor, shows impressive results in the treatment of CLL, and efficacy has also been demonstrated in DLBCL. Methods: We report on a 48-y-old female pt. diagnosed with CLL in 5/2014 with leukocytosis of 50.000 with 80% lymphocytes and generalized lymphadenopathy. Bone marrow (BM) revealed a CD5/CD23/CD79b/ CD22-positive, FMC7-negative B-cell population with 11q-deletion. In 2008 the pt. had suffered from gastric DLBCL, stage IIIA, IPI 1, successfully treated (CR) with 6 cycles of R-CHP and 2xR. Vincristine had not been given because of underlying hereditary sensorimotor neuropathy. A lymph node biopsy, performed at this time, showed CLL infiltration, clonally related in PCR-analyses to CLL in BM, however, not with the former gastric DLBCL. Because of symptomatic lymphadenopathy 6 cycles of R-FC were given with ensuing CR. Only 4 months later pt. presented with rapidly enlarging lymph nodes, elevated WBC (lymphocytosis 60%) and rising LDH. Results: BM biopsy showed a 50% infiltration of lymphoid cells with CLL profile and 17p deletion. Due to progressive cervical lymph node swelling and elevated LDH a re-biopsy was done revealing now DLBCL that was clonally related with CLL diagnosed in May 2014, qualifying for Richter’s syndrome. Salvage chemotherapy with R-ICE was initiated. After one cycle, lymph nodes were reminiscent and the pt. developed pancytopenia, B-symptoms and a once again rising LDH. BM biopsy revealed 90% infiltration of blastoid CD45 dim positive cells suspicious for a more aggressive transformed population also in BM. In this chemo-refractory situation allogeneic SCT was scheduled. To achieve remission ibrutinib was given at a dose of 420mg daily. After 8 weeks of therapy LDH normalized, and the CT scan showed good PR (-59%). The pt. is currently awaiting allogeneic SCT. Conclusion: Ibrutinib shows good and rapid response in a patient with Richter’s syndrome. Disclosure: Angela Fischer-Maranta: No conflict of interest disclosed. Karin Hohloch: Financing of Scientific Research: Bayer, Roche, Spectrum; Expert Testimony: Spectrum

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P535

Severe secondary hemophagocytic lymphohistiocytosis (HLH) concomitant with diffuse large B-cell lymphoma of the spleen Parmentier S.1, Ott G.2, Ehleiter H.1, La Rosée P.3, Schaich M.1 Rems-Murr-Kliniken Winnenden, Hämatologie, Onkologie, Palliativmedizin, Winnenden, Germany, 2Robert-Bosch-Krankenhaus, Abteilung für Klinische Pathologie, Stuttgart, Germany, 3Universitätsklinikum Jena, Klinik für Innere Medizin II, Abt. für Hämatologie und internistische Onkologie, Jena, Germany 1

Introduction: HLH is a rare syndrome characterized by extreme immune activation, resulting in pathologic inflammation. The diagnosis of HLH may be established by a molecular diagnosis consistent with HLH or five of 8 criteria consisting of fever, splenomegaly, cytopenias, hypertriglyceridemia, hemophagocytosis in bone marrow, spleen, lymph nodes or liver, low NK-cell activity, high ferritin levels and/or elevated soluble CD25. “Primary” HLH describes infants with predisposing inherited immune deficiencies, and “secondary” HLH describes older patients in whom significant immune activation may be caused by e.g. autoimmune disease, persistent infection or malignancy. Case report: A 32 year old male patient presented to our department for further evaluation of intermittent fever and progressive neutropenia. Infectious causes could be excluded. Bone marrow aspirate revealed reactive changes and hemophagocytosis. Laboratory work-up revealed elevated liver enzymes, hypofibrinogenemia, hypertriglyceridemia, massive elevated ferritin levels of 16.422 mg/l, LDH 1772 U/L and a sIL2-receptor of >7200 U/ml. CT scans showed an enlarged liver and spleen and retroperitoneal lymphadenopathy. No skin eruptions, further lymphadenopathy or joint pain was described. Within the spleen, multiple hypodense nodules were seen. In a retroperitoneal lymph node the diagnosis of HLH was suggested. Meanwhile, the patient´s condition worsened with signs of capillary leak mainly due to the massive inflammation and hypalbuminemia. Therefore, treatment with dexamethasone (according to HLH2004 trial protocol) and etoposide (75 mg/m²/week) was started. We then conducted a splenectomy where the final diagnosis of a diffuse large B-cell lymphoma with features of T-cell/histiocyte large B cell lymphoma was made. Cytotoxic treatment with the R-CHOEP protocol was initiated. The patients condition improved rapidly. Ferritin, sIL2R and liver enzymes declined accordingly. Follow-up of the patient will be presented. Conclusion: HLH in adults is a life-threatening syndrome. Often, immediate antiinfllammatory treatment with the adapted pediatric protocol is mandatory to prevent irreversible organ damage. Yet, diagnosis of the underlying trigger disease is pivotal to effectively overcome the detrimental immune stimulation by disease specific treatment. Disclosure: No conflict of interest disclosed.

Posterdiskussion Multiples Myelom 2 P536

The combination of novel agents and high cut-off dialysis is crucial in multiple myeloma patients with acute kidney injury Gerth U.1, Görlich D.2, Thölking G.1, Berdel W.E.3, Pavenstädt H.1, Kümpers P.1, Pohlen M.3 Universitätsklinikum Münster, Medizinische Klinik D – Innere Medizin, Nephrologie und Rheumatologie, Münster, Germany, 2Universitätsklinikum Münster, Institut für Biometrie und Statistik, Münster, Germany, 3 Universitätsklinikum Münster, Medizinische Klinik A – Hämatologie und Onkologie, Münster, Germany 1

Introduction: Although multiple improvements in novel chemotherapeutic agents emerged in the last decades and improved therapy response of multiple myeloma (MM) patients with an acute kidney injury (AKI) still have a poor prognosis. AKI often results from high concentrations of circulating monoclonal serum free light chains (sFLC) which can induce a tubular-interstitial damage and might become irreversible if sFLC

Abstracts

concentrations are not reduced rapidly. In addition to application of novel chemotherapeutic agents, reduction of high sFLC concentrations can be achieved by high cut-off hemodialysis (HCO-HD). The aim of this study was to analyze the effects of both novel chemotherapeutic agents and HCO-HD in MM patients with AKI. Methods: We retrospectively analyzed 59 MM patients suffering from AKI at time of hospital admission. They were treated with / without novel agents (36 vs. 23 patients) and with / without HCO-HD (42 vs. 17 patients), respectively. Outcome parameters were a) sustained decline in sFLC-values, b) recovery of renal function and c) 1-year OS. Results: A rapid and sustained decrease in sFLC values was associated with an increased rate of renal recovery (p = 0.013). In addition, also Kaplan-Meier curves display a trend towards increased one year OS (p = 0.044). In patients treated with HCO-HD, a sustained sFLC response was observed more often (83.3% vs. 29.4% without HCO-HD, p = 0.007). In addition, also the rate of renal recovery was significantly higher (64.3% vs. 29.4%, p = 0.014). The type of chemotherapeutic agent significantly influenced renal recovery (p = 0.015). In detail, patients treated with novel agents showed a renal recovery rate of 50.0% (non HCO-HD) which could be increased to 65.6% by HCO-HD (p = 0.540). In contrast, if treated with protocols excluding any novel chemotherapeutic agents, the corresponding rates of renal recovery were 23.1% compared to 60.0% with HCO-HD (p = 0.072), respectively. Conclusions: A rapid and sustained decline in sFLC values is crucial in therapy of MM patients with AKI. Decline in sFLC values predicts recovery of kidney function and might also be essential for OS. The combination of novel chemotherapeutic agents and HCO-HD show additional benefits on a rapid sFLC removal and an improved renal outcome. Disclosure: No conflict of interest disclosed. P537

Pattern of organ damage at presentation and relapse after autologous stem cell transplantation (ASCT) in patients with multiple myeloma – influence of functional parameters Arat P.1, Teutloff C.1, Liesenjohann S.1, Baier J.1, Gerrlich C.1, Boquoi A.1, Strapatsas T.2, Wilk M.3, Kondakci M.1, Kobbe G.1, Haas R.1, Fenk R.1, Neukirchen J.1 Klinik für Hämatologie, Onkologie und klinische Immunologie, Universitätsklinikum Düsseldorf, Düsseldorf, Germany, 2Helios Klinikum Wuppertal, Wuppertal, Germany, 3Universitätsspital Zürich, Zürich, Switzerland 1

Introduction: High-dose chemotherapy and ASCT is currently the gold standard of first-line treatment in patients with multiple myeloma without significant comborbidities. We investigated whether the pattern of organ damage has an impact on outcome besides known prognostic parameters like ISS stage and cytogenetics. Methods: We evaluated the clinical course of 291 patients with MM with a median age of 58 years (range 30–75 yr) treated with ASCT as a firstline treatment at our institution between Jan 2006 and June 2014. Clinical characteristics as CRAB criteria at diagnosis and at the time of progression were analysed with regard to progression free (PFS) and overall survival (OS). Results: At the time of diagnosis, 20% of the patients were classified as ISS stage III. Hypercalcemia, creatinine >2 mg/dl, anaemia with Hb< 10g/ dl and osteolytic bone lesions were found in 9%, 12%, 30% and 85%, respectively. Elevated creatinine and anaemia were associated with an unfavourable OS (p = 0.019 and p = 0.048). Induction therapy included bortezomib in 62%. 69% of the patients received a maintenance therapy, which mostly consisted of lenalidomide (62%). After a median follow up of 38 months, the median PFS and OS were 36 and 94 months, respectively. Parameters predicting outcome significantly estimated by univariate analysis are shown in table 1.

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Tab. 1. Parameters affecting progression and survival

ISS III Age above 65 Extramedullary disease Radiotherapy No bortezomib containing induction No maintenance therapy No CR as best response

n 55 61 74 126

HR PFS 1.443 0.803 1.251 1.891

p 0.070 0.299 0.235 <0.0001

HR OS 1.680 1.0002 2.252 2.111

p 0.056 0.995 0.001 0.003

101

1.507

0.016

1.143

0.579

73 82

2.068 2.216

<0.0001 3.008 <0.0001 3.748

<0.0001 <0.0001

In multivariate analysis, GFR< 50%, extramedullary disease, radiotherapy, no maintenance therapy and no CR were significantly associated with a worse outcome (p < 0.001). Conclusions: Our results suggest that significant impairment of organ function as anaemia and destructive bone lesions necessitating radiotherapy reflect a particular more aggressive type of disease with shorter PFS and OS. Further, we confirm the role of maintenance therapy as an important component of a HDT regime significantly improving PFS and OS. Moreover, our data confirm that age of 65+ is not a parameter to define transplant ineligibility. Disclosure: Pia Arat: No conflict of interest disclosed. Judith Neukirchen: Other Financial Relationships: Reisekostenunterstützung Celgene und BMS P538

Prospective comorbidity and functional geriatric assessment (CF-GA) in multiple Myeloma (MM) patients (pts): Results from a multicenter German study group MM (DSMM) trial Dold S.M.1, Zober A.1, Ihorst G.2, Pönisch W.3, Mügge L.-O.4, Knop S.5, Langer C.6, Schumacher M.7, Duyster J.1, Wäsch R.1, Engelhardt M.1 University of Freiburg Medical Center, Hematology & Oncology, Freiburg, Germany, 2University of Freiburg Medical Center, Clinical Trials Unit, Freiburg, Germany, 3University of Leipzig Medical Center, Hematology & Oncology, Leipzig, Germany, 4University of Jena Medical Center, Hematology & Oncology, Jena, Germany, 5University of Würzburg Medical Center, Hematology & Oncology, Würzburg, Germany, 6University of Ulm Medical Center, Hematology, Oncology & Rheumatology, Ulm, Germany, 7University of Freiburg Medical Center, Institute of Medical Biometry and Medical Informatics, Freiburg, Germany 1

Introduction: Cancer pts present with a highly heterogeneous health status and treatment choices are numerous. Therefore, careful assessment of individuals´ condition is relevant. In order to define best tolerable treatment options, novel metrics for non-disease variables are needed. Albeit impairment of the Karnofsky Performance Status (KPS), Activities of Daily Living (IADL or ADL) and Quality of Life (QoL) are predictive for outcome in cancer pts, the prognostic variables within a defined and prospectively assessed battery of established functional tests have rarely been assessed in MM. Methods: In this trial, we performed a prospective CF-GA in MM pts prior to initiation of anti-myeloma treatment and reflected pts´ baseline health status. It included the IADL, ADL, Timed Up and Go Test (TUGT), malnutrition, pain, fitness, SF12-QoL and geriatric depression scale (GDS). Moreover, established comorbidity (CM) scores like ß2MG/ eGFR, Kaplan Feinstein (KF), HCT-CI, CCI, IMWG score, initial-Myeloma Comorbidity Index (I-MCI) and revised-MCI (R-MCI) were assessed. The CF-GA was performed as a screening tool to assess pt fitness as well as to predict survival. Results: Overall, 259 pts have currently been included: these were typical for tertiary centers with a median age of 62 years (27–84). The median hemoglobin was 11.0g/dl (5–17), eGFR 71ml/min/1.73m2 (7–163), ß2MG 3.4mg/l (0.8–38.4) and BM infiltration 40% (0–95). The frailty assessment revealed a median KPS of 80% (30–100) and subjective fitness was graded with 3 (1–6). Median objective results for the IADL were 8 (1–8), for ADL 5 (2–6), for pain 2 (0–10), for malnutrition 4 (0–14), for MMSE 28

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(15–30), for GDS 2 (0–13) and for TUGT 10 (4–80). Median CM scores were substantially different with an I-MCI of 1 (0–3), ß2MG/eGFR of 1 (0–2), KF of 1 (0–3), CCI of 2 (0–8), HCT-CI of 2 (0–9) and R-MCI of 4 (0–9). In age subgroups, mean R-MCI and IMWG did increase the most and much lesser with HCT-CI and KF, confirming the latters´ lesser usefulness in MM. Conclusion: To the best of our knowledge, this is the first prospective multicenter GA that aggregates most valuable MM-specific disease risks, frailty tests and CM scores. Our current results in 259 pts showed that the most relevant and compromised parameters in MM are: KPS, TUGT, ADL, R-MCI and IMWG score. Most predictive CF-GA tools should be included in future clinical trials. Disclosure: No conflict of interest disclosed. P539

“Routine assessment of patient-reported outcomes (PROs) in patients with multiple myeloma” An analysis of the Austrian Myeloma Registry (AMR) Weger R.1, Willenbacher W.1,2, Loth F.3, Willenbacher E.2, Blazek T.3 Oncotyrol – Center for Personalized Cancer Medicine, Innsbruck, Austria, Department of Hematology & Oncology, Innsbruck Medical University, Internal Medicine V, Innsbruck, Austria, 3Department of Psychiatry and Psychotherapy, Innsbruck Medical University, Innsbruck, Austria 1 2

Background: Cancer patients suffering from multiple myeloma, a chronic disease with multiple treatment options, experience a variety of disease or treatment-related symptoms. Frequently experienced problems including physical and emotional impairments and the economic burden of the disease are known to have an adverse effect on patients’ quality of life. Routine assessment of patient-reported outcomes (PROs) to monitor the impact of the disease and its treatment allows guiding specific medical and psychosocial interventions. Patients and methods: To gain a comprehensive picture of the patient’s current health status and well-being, we extended the Austrian Myeloma Registry (AMR) with routine PRO assessments. Using the software CHES (Computer Based Health Evaluation System) we collect data on essential outcome parameters covered by internationally widely used quality of life questionnaires (EORTC QLQ-C30 and QLQ-MM20). The electronic data collection infrastructure allows collecting PROs in in- and outpatients at home or in day care centres. This data can be made immediately available to the treatment team using sophisticated longitudinal graphical presentations to support medical decision making. At present we have included more than 100 multiple myeloma patients in the routine PRO assessment for the AMR. First results will be shown at the conference. Conclusion: PROs have become important outcome parameters for the evaluation of the treatment of multiple myeloma. They allow assessing the impact of disease and treatment on quality of life and capturing specific physical symptoms (including side-effects). Including the patient’s individual perspective that is directly reported by him/herself in cancer registers is essential to obtain a comprehensive picture of the benefits and burden caused by the various available treatment options. The use of such data allows to improve shared medical decision making and contributes to patient empowerment. Disclosure: No conflict of interest disclosed.

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P540

Diagnosis and treatment of multiple myeloma in Germany – Analysis of a nationwide, multi-institutional survey Merz M.1, Kellermann L.2, Poenisch W.3, Tischler H.-J.4, Kohnke J.5, Knauf W.6, Goldschmidt H.1 Universitätsklinikum Heidelberg, Hämatologie, Onkologie und Rheumatologie, Heidelberg, Germany, 2OncologyInformationService, Freiburg, Germany, 3 Universitätsklinikum Leipzig, Leipzig, Germany, 4MKK- Johannes Wesling Klinikum Minden, Minden, Germany, 5Onkologische Praxis Remscheid, Remscheid, Germany, 6Onkologie Bethanien, Frankfurt am Main, Germany 1

Introduction: With this update of a nationwide, multi-institutional survey we analysed whether treatment reality outside clinical trials reflects current advances in the management of multiple myeloma (MM). Methods: University hospitals (UH), community hospitals (CH) and office-based haematologists (OH) were contacted in 2011 and 2015 to provide data from MM patients treated for newly diagnosed or relapsed disease during the first half of the respective year. Data were extracted from patient records and entered into an online database. We identified 478 (2011) and 515 (2015) that were treated at CH (2011: 40% / 2015: 43%), UH (2011: 20% / 2015: 11%) and OH (2011: 40% / 2015: 47%). (multiple answers in case of mutual treatment) Results: Median age at primary diagnosis increased from 69 (2011;n = 478) to 71 years (2015;n = 515). Patients were diagnosed less frequently with concomitant diseases (2011: 61%/ 2015: 51%) and no changes in performance status were observed (ECOG 0-1 2011: 66%/ 2015: 68%). Cytogenetic analysis for risk assessment slightly increased from 2011 (53%) to 2015 (59%), with different acceptance among health care providers (UH: 68%/ CH: 58%/ OH: 56%). Factors associated with less frequent testing in frontline setting were age above 70 years or ineligibility for autologous stem cell transplantation (ASCT; p = 0.001, respectively). Number of patients ≥ 65 years increased from 2011 (27%) to 2015 (57%) among candidates for ASCT. Bortezomib-based therapies were used in 92% of transplant-eligible and 66% of transplant-ineligible patients in 2015. Treatment of relapsed disease changed from 2011 to 2015. In 2011, patients in first relapse were treated with bortezomib in 45% and lenalidomide in 27%. This ratio reversed in 2015, since 28% of patients in first relapse received bortezomib and 54% lenalidomide. After approval of pomalidomide for relapsed MM, 11% of patients in second relapse received the respective agent in 2015. Usage of lenalidomide for second relapse decreased from 2011 (36%) to 2015 (23%). Conclusion: In an aging MM population, we observe increasing acceptance of ASCT above 65 years, most likely due to preserved performance status and less comorbidities. Bortezomib-based induction therapies are standard of care for front-line therapy in Germany, especially in transplant-eligible patients. Lenalidomide is mainly used in first and second relapse, while with pomalidomide, the first second generation novel agent entered MM treatment in 2015.

as reference, 1 university hospital. From March 2014 until August 2015, patients fulfilling all following criteria were documented: 1) verified diagnosis of multiple myeloma; 2) last visit within 12 months; 3) 1–2 previous therapies; 4) presence of relapse or non-response to therapy; 5) no commence of 4th line therapy. Documentation included patient characteristics, anamnestic and laboratory as well as clinical parameters at diagnosis and present, and previous and current therapeutic strategies. Data were categorized for e.g. age, time since diagnosis and severity of disease. Results: In total, data of 378 patients were documented by office based hematologists, 50 patients by the clinic. A preliminary analysis for patients treated by office based hematologists was carried out. Overall, 45% of patients were female and 55% male. At diagnosis, nearly half of the patients (46%) were >70 years and 18% were between 66 and 70 years. Only a minority of patients was younger than 50 years (5%). Multiple myeloma was mainly diagnosed 1–5 years ago (56%; 6–10 years: 24%) and the frequency of visits to the physician was high (≥ every 3 months) in most patients (83%). Besides chemotherapy/steroids, most patients received Bortezomib 1st line treatment (71%), while patients in the 2nd or 3rd line were treated with either Bortezomib (34% and 30%, respectively) or immunomodulatory drugs (37% and 44%, respectively). Patients treated in the clinic differed from those described above (e.g. age, clinical study participation). A comparison between the 2 populations is currently being evaluated. Conclusion: According to the preliminary retrospective data analysis, Bortezomib-based regimens dominated the 1st line treatment of MM. For the 2nd and 3rd line treatment, no such clear preference was observed. A final analysis of the data will be presented at the conference, including a comparison between patients treated by office based hematologists and patients treated in a university hospital. Disclosure: Marc on behalf of the MM-Retro Members: Raab: Advisory Role: Novartis, Amgen, Bristol-Myer Squibbs; Expert Testimony: Novartis, Amgen, MorphoSys P542

Neuropathy in monoclonal gammopathies – when should the underlying disease be treated? Riedel K.1,2, Yomade L.O.2, Brioli A.2, Hochhaus A.2, Mügge L.-O.2 Sophien- und Hufeland-Klinikum gGmbH, Klinik für Gynäkologie und Geburtshilfe, Weimar, Germany, 2Universitätsklinikum Jena, Klinik für Innere Medizin II, Jena, Germany 1

Introduction: Treatment options for relapsed/refractory multiple myeloma (RRMM) have increased significantly over the last years and more will become available in the near future. The current analysis of a national survey was undertaken to gain insights into the disease management of RRMM before the market introduction of a variety of novel therapies. Methods: A retrospective data collection using questionnaires was performed by 18 office based hematologists distributed all over Germany and,

Introduction: In monoclonal gammopathies like multiple myeloma, Waldenström´s disease, MGUS and others, approximately one third of all patients report neuropathic symptoms. However, there are no clear cut criteria for defining when such symptoms should initiate treatment of the underlying gammopathy. We sought to extract parameters from clinical examination and nerve conduction studies which allow to reproducibly define the need for treatment of the underlying gammopathy after emergence of neurologic symptoms. Methods: We clinically examined 100 patients diagnosed with either MGUS, multiple myeloma or Waldenström´s Disease, as well as 19 healthy volunteers according to a standardized neurologic examination protocol. Results were graded according to published neurologic scores: Neuropathy Symptom Score (NSS), Neuropathy Deficit Score (NDS) and Inflammatory Neuropathy Cause and Treatment Sensory Sumscore (ISS). Furthermore, all patients and healthy volunteers were subjected to nerve conduction studies of the median and common peroneal nerves, as well as to a more advanced electrophysiological test method, the “collision technique”. Results: Since the clinical scoring systems NSS, NDS and ISS showed poor sensitivity and specificity in the target patient population, we developed the “Gammopathy Neuropathy Score” (GNS). This score showed improved sensitivity and specificity in identifying patients with monoclonal gammopathies and associated neuropathy. A score value above 2 points (out of 24) was able to predict measurable neuropathy with a sensitivity of 75.0% and a specificity of 70.3%. Test results of the GNS were correlat-

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–160

Disclosure: Maximilian Merz: No conflict of interest disclosed. Hartmut Goldschmidt: Advisory Role: Celgene, Janssen; Financing of Scientific Research: Celgene, Janssen; Expert Testimony: Celgene, Janssen P541

Retrospective analysis of the disease management of patients with relapsed and/or refractory multiple myeloma after 1–2 previous therapies Raab M.S.1, on behalf of the MM-Retro Members Universitätsklinikum, Heidelberg, Germany

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ed with the results of nerve conduction studies. The conduction study of the peroneal nerve proved to be useful in identifying patients with sensory and motor nerve damage due to gammopathies. Surprisingly, even in patients with “monoclonal gammopathy of undetermined significance” (MGUS), some degree of neuropathy could already be detected in comparison to healthy volunteers. In addition, the collision technique was able to better differentiate between gammopathy patients with minor neurological damage and patients not affected by neuropathy. Conclusion: A standardized workup including clinical examination, scoring by GNS and nerve conduction studies in patients with a score value above 2 will help in identifying patients that should get treatment of the gammopathy to prevent further neurological damage. Disclosure: No conflict of interest disclosed. P543

VDCR as a treatment option for refractory or relapsed multiple myeloma in Bortezomib pretreated patients, a single center experience Dobrosch L.1, Hahn-Ast C.1, Mayer K.1, von Lilienfeld-Toal M.1, Brossart P.1, Janzen V.1 Universitätsklinikum Bonn, Medizinische Klinik III, Bonn, Germany

Introduction: Current standard of care in newly diagnosed multiple myeloma includes at least one of the “novel agents” , i.g. a proteasome inhibitor(PI) or immunomodulatory drug (IMiDs). However, a significant proportion of patients are refractory to initial therapy and most patients will eventually relapse or become refractory to any established treatments over time and will need experimental approaches to overcome drug resistance. Methods: This is a single center retrospective study of 24 RRMM patients who received a four drug combination therapy with bortezomib, lenalidomide, cyclophosphomide and dexamethasone (VDCR) between November 2010 and December 2015 (some in cooperation with associated centers upon our recommendation). Response to VDCR treatment was evaluated based on IMWG criteria in terms of reduction in paraprotein secretion. In addition, the survival is being evaluated. Results: Of the 24 Patients, 12 were female. Two patients received VDCR-protocol twice and were thus analyzed twice (one man, one woman), counting 26 cases. The average age of patients starting VDCR was 61.4 years. 10 patients were also lenalidomide pretreated. 14 patients had more than two previous therapies. In 24 of 26 cases, at least a partial response (≥50% reduction of paraprotein) was achieved after VDCR treatment. In 11 cases, a reduction of ≥ 90% was observed, equivalent to VGPR. The average paraprotein reduction was 81.2%. The follow up for survival is still ongoing. The 12 month survival rate was 68%. Conclusion: Within its limitations as a retrospective single center, this analysis shows that VDCR treatment is a highly effective combination even in heavily pretreated and refractory myeloma patients.

for RRMM. We report preliminary data on HCRU on the overall study population (North America and Europe). Methods: Patients (pts) aged ≥18 yrs with RRMM (≥1 prior therapy) who initiated Tx with an IMiD, PI, or IMiD+PI 90 days prior/30 days after study enrolment were eligible. Pt data were collected at each healthcare provider (HCP) visit, including number and reason for clinic/physician office visits, home healthcare, hospital outpatient and emergency room visits, and hospitalizations. Results: At data cut-off (Dec 2015), 764 treated pts (median age 68 yrs, 55% male) were enrolled; 599 (79%) relapsed MM, 161 (21%) refractory MM. Lines of prior Tx: 1 (n = 325, 43%), 2 (n = 203, 27%), 3 (n = 111, 15%) and >3 (n = 123, 16%). Median (range) time from diagnosis to index MM Tx: 41 mths (24–71). Prior transplantation experience: 365/762 (48%) pts. At enrolment, 48% (n = 368) received an IMiD (lenalidomide 81%, n = 297), 45% (n = 347) a PI (bortezomib 80%, n = 278) and 6% (n = 49) an IMiD+PI; median (Q1-Q3) follow-up was 15.1 mths (8–24). Median (Q1-Q3) number of HCP visits/pt in Yr 1 was 4.5 (0–20) and 7 (0–20) per refractory pt. Median (Q1-Q3) number of HCP visits in Yrs 2 and 3 was 2 (0–12). Types and reasons for HCP visits, by prior lines of Tx, are shown in the Table. Of 6804 total HCP visits, 44% were hospital outpatients, 41% clinic/physician and 5% hospitalizations. MM management was the main reason given for 90% of HCP visits and 66% of hospitalisations. Conclusions: Regardless of prior Tx, routine management and disease progression constitute the majority of MM-related HCRU. Early use of novel Tx with potential to provide durable responses could improve MM outcomes. Cost analyses are ongoing. Study Funding: Bristol-Myers Squibb (BMS). Professional writing assistance by K Rees of Caudex, funded by BMS Tab. 1. Visits to HCP by Number of Prior Lines of Therapy

Disclosure: No conflict of interest disclosed. P544

Healthcare resource utilization (HCRU) in relapsed/refractory multiple myeloma (RRMM): results from PREAMBLE Goldschmidt H.1, Vij R.2, Kuter D.3, Cella D.4, Zyczynski T.5, Davis C.5, Popov S.6, Cook G.7 Universitätsklinikum Heidelberg, Medizinische Klinik V, Heidelberg, Germany, 2Washington University School of Medicine, St. Louis, United States, 3Massachusetts General Hospital Cancer Center, Boston, United States, 4 Northwestern University, Evanston, United States, 5Bristol-Myers Squibb, Princeton, United States, 6Parexel, St. Petersburg, Russian Federation, 7University of Leeds, Leeds, United Kingdom 1

Disclosure: Harmut Goldschmidt: Advisory Role: Bristol-Myers Squibb, Celgene, Janssen, Novartis, Chugai, Onyx, Millennium, Takeda, Amgen; Financing of Scientific Research: Bristol-Myers Squibb, Celgene, Janssen, Novartis, Chugai, Onyx, Millennium, Takeda, Amgen; Expert Testimony: Bristol-Myers Squibb, Celgene, Janssen, Novartis, Chugai, Onyx, Millennium, Takeda, Amgen Gordon Cook: Advisory Role: Celgene, Bristol-Myers Squibb, Janssen, Takeda, Chugai, Amgen, Jazz Pharma; Expert Testimony: Celgene; honoraria: Celgene, Bristol-Myers Squibb, Janssen, Takeda, Chugai, Amgen, Sanofi; Other Financial Relationships: Speakers Bureau: Celgene, Janssen, Takeda, Chugai, Amgen, Sanofi

Introduction: PREAMBLE (Prospective REsearch Assessment in multiple Myeloma: an oBservationaL Evaluation; NCT01838512) is an international cohort study on the real-world effectiveness of immunomodulatory drugs (IMiDs), proteasome inhibitors (PIs) and combination therapy (Tx)

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P545

P546

Clinical tumor database for multiple myeloma

Randomized comparison of pegylated IFN-a2b versus IFN-a2b maintenance following tandem autologous transplant in multiple myeloma

Bittrich M.1, Krebs J.2, Fette G.2, Einsele H.1, Knop S.1, Puppe F.2 Universitätsklinikum Würzburg, Würzburg, Germany, 2Julius-MaximilianUniversität Würzburg, Würzburg, Germany 1

Multiple myeloma faces physicians with enormous challenges. Although the incidence in Germany is not definitely increasing, the prevalence of multiple myeloma is expected to raise due to an ageing society and improvements in the overall survival of affected individuals. With high socioeconomic impact, the epidemiology and therapy is worth having a closer look at it. Moreover, therapeutic regimes are in a constant state of change. Therefore it is necessary to analyze the patients’ characteristics and response to past and present therapies in detail. The University hospital of Wuerzburg is a referral center for the treatment of multiple myeloma in Germany. In 2014, 2.400 MGUS, smouldering myeloma, multiple myeloma, solitary plasmacytoma or plasma cell leukemia patients were followed up in the in- and outpatient clinic. The aim of the current project is to build up a clinical tumor database as new approach in epidemiological research at University Hospital of Wuerzburg. A clinical database containing information from patients’ characteristics to therapy, response and complications was to be established. Unfortunately, large amounts of clinical data are written in text (e.g. physician’s letter). So it’s impossible to calculate a statistic without proper preprocessing. With a new approach of our cooperation partner (Department for Artificial Intelligence and Applied Computer Science of the University of Wuerzburg) it is possible to extract and preprocess relevant information into a “clinical data warehouse”. All data are stored pseudonymized protected by a firewall. Using the existing enormous pool of data and knowledge for assessment and improvement of special issues in therapy of plasma cell diseases will be promising. Following trends in therapy gives a future-oriented structure and opens new horizons in physicians’ daily practice and background knowledge as well as quality assurance aspects. All patients suffering from plasma cell diseases treated at the University Hospital of Wuerzburg will be included in the database. The clinical data will contain extraction of patient’s individual characteristics (e.g. genetic markers), initial clinical parameters and their longitudinal development, diagnostics, treatment concepts, adverse events, relapse, time to relapse, and survival rates. This way it will be possible to derive scientific benefit from the large data pool of the University Hospital of Wuerzburg. Disclosure: No conflict of interest disclosed.

Schreder M.1, Liebisch P.2, Hebart H.3, Holler E.4, Engelhardt M.5, Metzner B.6, Peest D.7, Aulitzky W.8, Straka C.9, Fischer T.10, Sezer O.11, Hentrich M.12, Ostermann H.13, Peschel C.14, Heß G.15, Hertenstein B.16, Wolf H.-H.17, Jung W.18, Frickhofen N.19, Maschmeyer G.20, Langer C.21, Meisner C.22, Kanz L.22, Einsele H.23, Knop S.23, Deutsche Studiengruppe Multiples Myelom Universitätsklinikum Würzburg, Med. Klinik und Poliklinik II, Würzburg, Germany, 2Onkologische Praxis Moers, Moers, Germany, 3Stauferklinikum, Mutlangen, Germany, 4Universitätsklinikum Regensburg, Regensburg, Germany, 5 Universitätsklinikum Freiburg, Freiburg, Germany, 6Klinikum Oldenburg, Oldenburg, Germany, 7Medizinische Hochschule Hannover, Hannover, Germany, 8 Robert-Bosch-Krankenhaus, Stuttgart, Germany, 9Schön Klinik Starnberger See, Berg, Germany, 10Universitätsklinikum Magdeburg, Magdeburg, Germany, 11Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany, 12 Rotkreuzklinikum, München, Germany, 13Klinikum der Ludwig-Maximilians Universität, München, Germany, 14Klinikum rechts der Isar, München, Germany, 15 Universitätsmedizin Mainz, Mainz, Germany, 16Klinikum Bremen Mitte, Bremen, Germany, 17Universitätsklinikum Halle (Saale), Halle (Saale), Germany, 18 Universitätsmedizin Göttingen, Göttingen, Germany, 19Helios Kliniken Wiesbaden, Wiesbaden, Germany, 20Klinikum Ernst von Bergmann, Potsdam, Germany, 21Universitätsklinikum Ulm, Ulm, Germany, 22Universitätsklinikum Tübingen, Tübingen, Germany, 23Universitätsklinikum Würzburg, Würzburg, Germany 1

Introduction: Before the advent of more refined immunotherapies, interferon (IFN)-alpha 2b (α2b) was used in haematological malignancies to control states of lower disease activity. In multiple myeloma (MM), a large meta-analysis found some impact on both progression free (PFS) and overall survival (OS), but its use has been limited due to poor tolerability of the drug. The introduction of a pegylated IFN-α2b derivative (PEGIFN) associated with less toxicity led us to compare both compounds for maintenance after intensive first-line treatment. Patients and methods: In the German DSMM V trial, newly diagnosed MM patients (pts) up to the age of 60 years (yrs) without deletion of chromosome 13q (del13q) on fluorescence in situ hybridisation were considered to be of standard risk and included into this part of the trial. Following non-novel compound cytoreduction, pts underwent tandem high-dose melphalan 200 mg/m² and autologous stem cell transplantation (ASCT). Two to four months from the 2nd ASCT, pts were randomized to receive subcutaneous IFN-α2b up to 3 MU/m² three times a week or weekly PEG-IFN 50–80 µg depending on body weight until disease progression or the occurrence of unacceptable toxicities. The primary endpoint was the cumulative incidence of treatment discontinuations within two years from start of maintenance. Results: 135 pts with a median age of 53 (range, 30–61) yrs were enrolled. 65 pts were randomized to receive IFN-α2b (Arm A) and 70 to receive PEG-IFN (Arm B). Median follow-up for surviving pts is 51.0 (range, 23.8–152.3) months (mos). After two years, more than half of the pts had discontinued IFN maintenance with no difference between the arms (54% for Arm A vs. 64% for Arm B, p = 0.226). Drug-related events accounted for 34% vs. 43% of treatment discontinuations (p = 0.295). At the time of last follow-up, 59 pts (44%) had not experienced disease progression. Median PFS was 33 months and did not differ between treatment groups (33.3 vs. 33.9 mos, p = 0.261). Four years after randomization, 101 pts (75%) were still alive. Long term follow-up data will be presented at the meeting. Conclusion: The use of PEG-IFN for maintenance treatment did not improve tolerability and discontinuation rate compared to IFN-α2b. However, given the promising outcome in this group of non-del13q MM pts, IFN may be reconsidered in the context of novel immunoediting strategies such as monoclonal antibodies and checkpoint inhibitors. Disclosure: No conflict of interest disclosed.

Abstracts

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SIADH and epigastric infiltration by generalized varicella zoster infection Parmentier S.1, Ehleiter H.1, Engelhardt M.2, Schaich M.1 Rems-Murr-Kliniken Winnenden, Hämatologie, Onkologie, Palliativmedizin, Winnenden, Germany, 2Rems-Murr-Kliniken Winnenden, Allgemeine Innere Medizin, Gastroenterologie, Winnenden, Germany 1

Introduction: Varicella zoster virus infections in immunocompetent patients are often localized and self-limiting. In immunosuppressed patients it may become generalized and might be accompanied by severe organ failure. Therefore it should be recognized and treated early in the course of disease. Case report: A 72year old woman with multiple myeloma and second high-dose melphalan with autologous stem cell transplantation three months ago presented with severe epigastric pain, nausea and emesis. A gastroscopy revealed vesiculae, initially desribed as intramucosal bleeding. Her physical state worsened rapidly within 24 hours with somnolence due to severe hyponatriemia (112 mmol/l) and a platelet-drop from normal to 33 Gpt/l. She also developed fever up to 40°C and in the further course and multiple disseminated vesiculae all over the skin. High-dose aciclovir (3x 10 mg/kg i.v.) was started. Viral load in peripheral blood for varicella zoster (VZ) was 5 x 10^6 copies/ml. Histopathological results of the gastric vesiculae showed infiltrates of varicella zoster in the stomach. The patient recovered completely. Conclusion: Zoster infections are not rare in immunosuppressed patients. Skin and CNS are mainly affected. In rare cases, SIADH can be caused by increased release of ADH due to CNS involvement. Epigastric infiltration is not a typical organ affected by this virus which might cause difficulties in upfront diagnosis like in this case. Disclosure: No conflict of interest disclosed. P548

Confocal Laser Scanning Microscopy of corneal subbasal nerve plexus as a possible method to detect early signs of beginning peripheral neuropathy in multiple myeloma affected patients Koschmieder A.1, Kragl B.2, Diwoky S.2, Stachs O.1, Murua Escobar H.2, Guthoff R.1, Junghanss C.2 Department of Ophthalmology, University of Rostock, Rostock, Germany, Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany 1 2

Introduction: Patients with plasma cell disorders are frequently affected by peripheral neuropathy due to disease progression or therapeutic intervention. Currently, neuropathic progression is clinically monitored by symptoms of the patients and decreased sensation to vibration as tested the Rydel and Seiffer tuning fork. Herein we present the evaluation of a novel non-invasive microscopic method allowing to monitor neuodegenerative processes in cornea of multiple myeloma patients. We report on first results of a pilot study. In diabetes mellitus patients these changes precede peripheral neuropathy. Method: Multiple myeloma patients underwent an assessment of sensory nerve function of feet and legs by Rydell and Seiffer tuning fork, monofilament testing, temperature as well as achilles tendon reflex. Further they received unilateral non-invasive confocal laser scanning microscopy of the corneal subbasal nerve plexus and bilateral corneal sensitivity testing using Cochet-Bonnet Esthesiometry. In the pilot cohort first diagnosed patients as well as patients at different stages are evaluated. Results: The non-invasive confocal microscopy of the corneal subbasal nerve plexus offers distinct information about number, density and branching of peripheral nerve. Former studies dealing with diabetic peripheral neuropathy have shown a reduction of corneal nerve fibres detected with the same method even before the clinical detectable start of neuropathy symptoms and a correlation to the symptoms in the course of

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the disease. We expect to find a similar correlation in the study group of multiple myeloma patients. Conclusions: With the examination of corneal nerve density there is an established method available which can be used as a surrogate marker to monitor the neurotoxic effects of therapy in multiple myeloma patients even on a subclinical level. Disclosure: No conflict of interest disclosed. P549

Bisphosphonates or RANK-ligand inhibitors for patients with solid cancer and bone metastases or multiple myeloma: a systematic Cochrane review and meta-analysis Köhler N.1, Jordan K.2, Jahn F.2, Wörmann B.3, Skoetz N.1 Uniklinik Köln, Klinik I für Innere Medizin, Cochrane, Köln, Germany, Universitätsklinikum der Martin-Luther-Universität Halle-Wittenberg, Klinik für Innere Medizin IV, Halle, Germany, 3DGHO, Berlin, Germany 1 2

Introduction: In advanced solid cancer and multiple myeloma more than 50% of patients develop metastatic bone disease, with considerable impact of complications (e.g. bone pain, pathologic fractures, spinal cord compression). These complications can reduce quality of life and are associated with an increased risk of death. Therefore, preventing skeletal complications is an important goal in cancer patients. Bone-targeting agents like bisphosphonates and the inhibitor of RANK-ligand denosumab are known to reduce the development of skeletal complications. Denosumab is not approved for patients with multiple myeloma. The objective of this review is to compare the clinical benefits and harms of these agents. Methods: We searched Cochrane Central Register of Controlled Trials and MEDLINE (1950 to March 2016) for randomised controlled trials (RCTs) and included trials evaluating bisphosphonates in comparison to denosumab in patients with solid cancer and bone metastases or multiple myeloma. Two review authors independently screened search results, extracted data and assessed the quality of trials. We pooled trials using the random-effects model. Partly supported by German Cancer Aid (110645). Results: Five RCTs including 6089 patients were identified. Two RCTs (N = 2301) included breast cancer patients, one RCT (N = 1901) included prostate cancer patients and two RCTs (N = 1887) included patients with solid cancer and multiple myeloma. These five trials included only 189 patients with multiple myeloma (3%). Because data of these patients were not available we have not performed a separate analysis. For bone pain and quality of life the outcomes were too heterogeneous to pool in one meta-analysis. For skeletal-related events a meta-analysis could be performed and showed an advantage for denosumab (RR 0.86, 95% CI 0.79 to 0.93). In contrast, mortality (RR 1.03, 95% CI 0.95 to 1.11), adverse events (RR 0.99, 95% CI 0.98 to 1.00), severe adverse events (RR 0.98, 95% CI 0.93 to 1.04) and osteonecrosis of the jaw (RR 1.40, 95% CI 0.92 to 2.13) showed no evidence for a difference. Conclusions: Although five RCTs have been published, this review shows that the majority of outcomes were either too heterogeneous or showed no evidence for a difference between bisphosphonates and denosumab. Disclosure: No conflict of interest disclosed. P550

Cost analysis of patients with relapsed or refractory multiple myeloma (MM) based on real-world-evidence data of German oncological practices Feuerbach M.1, Freigang F.1, Schulte C.2, Hurtz H.-J.3, Schwarzer A.4, Lipp R.1 GermanOncology GmbH, Hamburg, Germany, 2Gemeinschaftspraxis Hämatologie-Onkologie, Dortmund, Germany, 3Gemeinschaftspraxis und Tagesklinik, Halle (Saale), Germany, 4Gemeinschaftspraxis für Hämatologie und Onkologie, Leipzig, Germany 1

Introduction: MM is one of the most occuring hematological malignancy and the therapy of these patients is cost-intensive for health insurances. This retrospective, observational study focused on the treatment of re-

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lapsed or refractory MM. The aim was to calculate the health utilization costs and to analyse the distribution of costs to the relevant cost drivers and stages of therapy. Methods: Data about antitumor medication, hospitalisation, concomitant medication, oncological services and other more were collected under real-world conditions in 26 German oncological practices. For the calculation of health utilization cost of patients with relapsed or refractory MM, it was determined to regard the interval of one year after the start of second line therapy. Additionally, further analyses were performed to calculate the costs between the 1st and 2nd as well as between the 2nd and 3rd progress. Therefore, all treatments and procedures in the database were transferred into German reimbursement systems for health care services. The mode of calculations was based on the methods of the IQWIG and represented direct costs for health insurances. Results: 261 patients with relapsed or refractory MM (49% women, 51% men, median age 75 years) were included in the statistical and cost´s analysis. The total cost of therapy for the first year after initiating a second line therapy were in mean 40,439.39 € per patient (90.4% for antitumor medication, 4.2% for hospitalisation, 3.1% for concomitant medication and 0.5% for oncological services). In contrast, the costs between 1st and 2nd progress were in mean 44,780.87 € (89.6% for antitumor medication, 3.7% for hospitalisation, 3.2% for concomitant medication and 1.8% for oncological services) whereas the costs between 2nd and 3rd progress come to 28,970.56 € (76.0% for antitumor medication, 12.6% for hospitalisation, 6.3% for concomitant medication and 2.7% for oncological services). Conclusions: In the first year after initiation of a second line therapy for MM the annual cost of therapy was calculated with a mean of 40,439 € per patient. Nearly 90% of the cost per patient would be expended for antitumor medications. Further analyses of costs between 1st and 2nd and between 2nd and 3rd progression assessed the fact that cost per patient during a second line was one third higher than during a third line therapy. Disclosure: No conflict of interest disclosed.

Posterdiskussion

Allogene Stammzelltransplantation 2

Results: Median time between first HCT and HHCT was 11 months (3–56 months) and 2 months (2–9 months) between relapse and HHCT. Time to relapse after HCT did not influence overall (OS) and disease free (DFS) survival (p = 0.54 and 0.45). In 2 patients, HHCT was performed as 3rd HCT 10 and 16 months after 2nd HCT, respectively. Disease status at HHCT was complete remission (CR) in 9 (35%) or active disease (bone marrow or peripheral blasts >5% or >0%, respectively) in 17 (65%) patients. Median neutrophil and platelet engraftment was noted on days 11 and 13, respectively without any patient experiencing full graft failure. No post-grafting immunosuppression was applied in case of < 5×104 per kilogram bodyweight T cells in the grafts. Otherwise, mycophenolate was applied. Low grade acute and chronic GvHD was observed in 12 (46%) and 5 patients (19%), respectively. GvHD did not contribute to overall mortality. Median OS was 6.7 months with 5 patients (19%) being alive at the end of follow-up. DFS was 6.0 months with a cumulative incidence of relapse of 49% at 12 months. In subgroup analyses, active disease at HHCT was an independent risk factor (multivariate analysis, both p = 0.017) for OS (3.4 vs. 9.0 months) and DFS (4.5 vs. 20.1 months). Long-term survival was very favorable for patients with CR (42% at 24 months). Conclusions: HHCT represents a curative treatment option for disease relapse after precedent HCT and features limited toxicity, favorable engraftment and low rates of GvHD. Especially patients with CR at HHCT have a promising prognosis. Hence, HHCT should be considered especially in these patients. Disclosure: No conflict of interest disclosed. P552

Underweight and obese patients – a challenge in allogeneic hematopoietic cell transplantation Haen S.P.1,2, Pham M.1, Faul C.1, Vogel W.1, Kanz L.1, Bethge W.A.1 Medizinische Universitätsklinik Tübingen, Abteilung II für Onkologie, Hämatologie, Immunologie, Rheumatologie und Pulmologie, Tübingen, Germany, 2Interfakultäres Institut für Zellbiologie, Abteilung Immunologie, Tübingen, Germany 1

Introduction: Disease relapse after allogeneic hematopoietic cell transplantation (HCT) is a serious therapeutic challenge. A second transplantation is associated with significant mortality. Haploidentical HCT (HHCT) is a therapeutic option since literally every patient has a haploidentical donor, and HHCT may induce more pronounced antileukemic effects through a greater HLA-disparity. Methods: 26 consecutive patients (9 women, 17 men, median age 37 years, 18–59 years) received HHCT upon disease relapse (ALL n = 7, AML n = 17, CML n = 1, PNH n = 1) after allogeneic HCT between 2000 and 2015. HHCT was performed as second (n = 24) or third (n = 2) transplantation. Haploidentical donors were parents (n = 6), siblings (n = 10), children (n = 5) or more distant relatives (n = 5). Grafts were manipulated with CD34+ selection (n = 5), CD3/CD19 (n = 18) or TCRαβ depletion (n = 3).

Introduction: Recent advances in conditioning regimens, supportive therapy and donor selection allows allogeneic hematopoietic cell transplantation (HCT) for an increasing number of patients with more complex risk profiles in need of adjustment to their individual conditions. Hence, patients with under- (UW) or overweight (OW) may require adjustment of conditioning dosages, graft cell counts and drug levels of immunosuppressive agents. Methods: 65 UW (body mass index – BMI < 20 kg/m²; 5%; median BMI 19.0 kg/m² (14.3–20.0 kg/m²)) and 90 OW (BMI > 30 kg/m², 7%, median BMI 32.7 kg/m² (30.0–49.4)) patients out of a cohort of 1,252 patients underwent allogeneic HCT (total of 161 HCT). UW patients were predominantly female (63%) and OW patients male (60%, p = 0.005), and were treated for acute (AML n = 28/53, ALL n = 14/5) and chronic (CMML n = 2/2, CLL n = 1/0, CML n = 0/1) leukemia, myelodysplastic (n = 4/14) and myeloproliferative (n = 4/3) syndromes, Hodgkin’s (n = 2/1) and non-Hodgkin (n = 9/6) lymphoma, multiple myeloma (n = 0/5), or histiocytic sarcoma (n = 1/0). Results: In 80% OW patients, chemotherapy dosages were limited to a body surface area of 2.3m². No adjustments were undertaken in UW patients. Hematopoietic engraftment (neutrophils >500/µl, platelets >20.000/µl) was noted on median day 18 (days 7–50) in both groups with 6 patients each without hematopoietic reconstitution. HCT-related toxicity was comparable with a tendency to more severe mucositis (p = 0.1) and kidney failure (p = 0.2) in the UW group. Patients were followed for a median of 30 months (0–135). Kaplan Meier estimated median overall survival (OS) for the whole cohort was 74 months with a median OS for UW and OW patients of 42 and 83 months, respectively (p = 0.4). Relapse rates were 38% and 33% (p = 0.8). Kaplan Meier estimate for median disease free survival (DFS) was 33 months with a median DFS for OW and UW patients of 20 and 47 months (p = 0.39). Cumulative incidence of non-relapse mortality (NRM) was both 25% for UW and OW patients.

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Haploidentical hematopoietic cell transplantation as a curative treatment option in patients experiencing disease relapse after precedent transplantation Haen S.P.1,2, Schumm M.3, Backert L.2,4, Löffler M.W.2,5, Faul C.1, Dörfel D.1, Vogel W.1, Handgretinger R.3, Kanz L.1, Bethge W.A.1 Medizinische Universitätsklinik Tübingen, Abteilung II für Onkologie, Hämatologie, Immunologie, Rheumatologie und Pulmologie, Tübingen, Germany, 2Interfakultäres Institut für Zellbiologie, Abteilung Immunologie, Tübingen, Germany, 3Universitätsklinik für Kinder- und Jugendmedizin, Abteilung I für Allgemeine Pädiatrie, Hämatologie und Onkologie, Tübingen, Germany, 4Eberhard-Karls-Universität Tübingen, Applied Bioinformatics Group, Tübingen, Germany, 5Universitätsklinik für Allgemeine, Viszeral- und Transplantationschirurgie, Tübingen, Germany 1

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Acute and chronic graft versus host disease (GvHD) was observed in 52 (OW/UW 24/28, p = 0.17) and 65 cases (OW/UW 39/26, p = 0.79), respectively. GvHD was mostly mild. Conclusions: In both UW and OW patients, allogeneic HCT can be safely performed with comparable survival and NRM. Although these results are similar to those in normal weight patients, dose adjustments of conditioning regimens should be considered to reduce toxicity especially in patients with significant OW. Disclosure: No conflict of interest disclosed. P553

Incipient bone marrow engraftment after aplasia during allogeneic and autologeous hematopoietic cell transplantation can be predicted by changes in uric acid serum levels Haen S.P.1,2, Eyb V.1,2, Mirza N.1,2, Vogel W.1, Faul C.1, Bethge W.A.1, Rammensee H.-G.2, Kanz L.1, Heni M.3 Medizinische Universitätsklinik Tübingen, Abteilung II für Onkologie, Hämatologie, Immunologie, Rheumatologie und Pulmologie, Tübingen, Germany, 2Interfakultäres Institut für Zellbiologie, Abteilung Immunologie, Tübingen, Germany, 3Medizinische Universitätsklinik Tübingen, Abteilung IV für Endokrinologie und Diabetologie, Angiologie, Nephrologie und Klinische Chemie, Tübingen, Germany 1

Introduction: Prolonged aplasia and graft failure represent potentially life-threatening complications for patients undergoing allogeneic and autologous hematopoietic cell transplantation (HCT). Therefore, the use of suitable biomarkers for early detection and differentiation between graft failure and poor graft function is worth pursuing. Uric acid (UA) is a strong immunological danger signal playing a pivotal role in innate and adaptive immunity. Methods: Laboratory results were analyzed in the following cohorts: 50 consecutive allogeneic HCT patients (26 women, 24 men, median age 48 years); 50 autologeous HCT patients (16 women, 34 men, median age 51 years); 50 leukemia patients (24 women, 26 men, median age 42 years). UA, creatinine, urea and differential blood counts were determined daily. Results: Mean baseline UA levels were 4.9, 5.5 and 3.9mg/dl in patients undergoing allogeneic HCT, autologeous HCT and induction chemotherapy, respectively. During therapy UA declined to mean values of 1.5, 2.1 and 1.7mg/dl (all p > 0.001) reaching hypouricemic values in most patients. After hematopoietic regeneration (neutrophils >500/µl) mean UA serum levels returned to baseline values (5.0, 4.2, 4.0mg/dl). Most patients undergoing HCT had no detectable leukocytes during aplasia while some leukocytes remained detectable after induction therapy. During aplasia UA levels remained low and started steadily increasing (defined as >2 consecutive days, median number of increases during aplasia n = 1) at a median of one day before rising leukocytes in allogeneic HCT (p = 0.01), and together with increasing leukocytes in autologous HCT (median increases n = 1). During induction chemotherapy, a steady increase of UA was also observed alongside rising leukocytes and neutrophils but also several times during aplasia (median 3 increases). UA serum levels and neutrophils followed similar kinetics with a statistical tendency to correlation (Pearson correlation 0.047). This indicated incipient immunological activity after HCT or remaining immunological activity during induction therapy. Of note, no increase of UA levels was observed without later rise of leukocytes. Conclusions: UA serum levels can serve as a sensitive biomarker for bone marrow activity and can indicate incipient release of neutrophils possibly reflecting consumption of early cells in the periphery. Therefore, UA serum levels can help to guide clinical differentiation between poor graft function and graft failure. Disclosure: No conflict of interest disclosed.

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Chronic GVHD as a risk factor for thrombembolism after allogeneic HSCT? Ries I.1, Hauptrock B.1, Schmitt T.1, Theobald M.1, Wagner E.M.1 UCT Universitätsmedizin Mainz, Hämatologie, internistische Onkologie, Pneumologie, Mainz, Germany 1

Introduction: Allogeneic hematopoietic stem cell transplantation (HSCT) is a curative treatment option for AML-patients, but up to 50% develop chronic graft versus host disease (cGVHD). Cardio-vascular morbidity such as heart failure, coronary heart disease and thromboembolic complications is also frequent (up to 23%, Savani Blood 2010). Mouse experiments showed endothelial damage after bone marrow transplantation and possible association between GVHD and vascular damage. Inflammation as part of the deregulated immune system in GVHD could promote vascular changes and increase the risk of cardiovascular complications. We asked if cGVHD and its inflammation are associated with thromboembolic complications. Methods: We retrospectively analyzed 414 patients with hematologic malignancies after allogeneic HSCT at our center (2009–2014). All patients were seen in an outpatient routine lifelong and received a GVHD screening according to the NIH consensus guidelines every 3 month (median follow up 3,8 years, range 0,8–6,7). Our patient cohort consists of 247 male and 167 female patients with a median age of 52,8 years. 50% suffered from AML (11% ALL, 10% lymphoma, 8% MPN, 7,5% MDS, 7% multiple myeloma). In 62,1% we used a dose reduced intensity conditioning regimen, in 27,3% a myeloablative regimen. 10,6% were treated with a FLAMSA-RIC. Donors were in 76,8% 10/10 matched unrelated donors (mainly peripheral blood stem cells). Results: 4,6% developed a new thromboembolic complication after HSCT. In 122 patients (27%) a chronic GVHD occurred, 59 with deep sclerotic phenotype. 8,9% of the patients with cGVHD developed a thromboembolic complication, mainly deep vein thrombosis, some with pulmonary embolism. In contrast, only 2,98% of patients without cGVHD showed a thromboembolic complication in the later time. We could not detect any relationship with other known risk factors or thrombosis bevor HSCT (1,8% with thrombosis before HSCT showed one afterwards). Conclusion: Our retrospective analysis showed an increased risk for thromboembolic complications after allogeneic HSCT, with substantial higher risk in patients with cGVHD (8,9%). In ongoing studies we investigate a vascular screening procedure with additional biomarkers according to inflammation and endothelial damage prospectively in patients with cGVHD. We hope to understand the individual vascular risks, identify patients at risk for thromboembolism and prevent future complications. Disclosure: No conflict of interest disclosed. P555

Central venous catheter-related complications in patients undergoing allogeneic or autologous hematopoietic cell transplantation or induction therapy for acute leukemia – A comparison between the internal jugular vein and the subclavian vein as insertion site Heidenreich D.1, Muetzel E.1, Kreil S.1, Nolte F.1, Faust M.1, Jawhar M.1, Hecht A.1, Hofmann W.-K.1, Klein S.A.1 Universitätsmedizin Mannheim UMM, III.Medizinische Klinik, Mannheim, Germany 1

Central venous catheters (CVC) are routinely used in patients undergoing hematopoietic cell transplantation (HCT) or induction chemotherapy (ICT) for acute leukemia. For these patients there are no data in the literature comparing the insertion sites internal jugular (IJV) or subclavian vein (SCV). Methods: All consecutive patients who were treated at our institution from 01/11 to 06/13 were included into this analysis. Inclusion criterion was insertion of a CVC due to allogeneic or autologous HCT or ICT for

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AML or ALL. CVC were inserted after disinfection by propanol/ethanol under full barrier precautions. The insertion site was chosen at the responsible physician´s own discretion. CVC were covered by chlorhexidine gluconate impregnated dressings. For analysis we built to groups, the allo-group and the non-allo-group (auto HCT and ICT). Results: 255 CVC were placed in 170 patients (median age: 59 years) due to allo-HCT (65 patients, 101 insertions), auto-HCT (30 patients, 33 insertions) or ICT (75 patients, 121 insertions). Underlying diseases were AML (102), ALL (9), MDS (15), MPN (4), lymphoma (21) or plasma cell neoplasia (19). Out of the 255 CVC 155 were sited via the IJV and 100 via the SCV. Among the 101 CVC in the allo-group 60 (60%) were placed in the IJV and 41 (41%) in the SCV [non-allo-group: 95 IJV (62%) vs. 59 SCV (38%)]. The median duration of catheterization per CVC for IJV and SCV was 18 vs. 17 days (d.) (ns) for the allo-group and 17 vs. 12 d. (ns) for the non-allo-group. The events fever or local infection occurred in 68% (41/60) of IJV- and in 93% (38/41) of SCV-CVC (p = 0.003) in the allo-group [non-allo-group: 79% (75/95) vs. 92% (54/59, p = 0.04). Median time to event was 12 (IJV) vs. 8d. (SCV, p = 0.005) for the allo-group [nonallo-group: 11 (IJV) vs. 9 d. (SCV, p = 0.0066). The data for the event local infection alone are also superior for the IJV: 29/60 (48%) vs. 29/41(71%) for SCV (p = 0.025) in the allo-group and 53/95 (56%) vs. 52/59 (88%) in the non-allo-group (p < 0.0001). Median time to local infection was 25 (IJV) vs. 12 d. (SCV, p = 0.01) in the allo-group and 15 (IJV) vs. 10 d. (SCV, p = 0.0004) in the non-allo-group. There were no significant differences in the frequency of BSI, thrombosis, pneumothorax or congestion of CVCs. No patient died CVC-related. Conclusions: In contrast to studies on non-hematologic patients these data demonstrate clearly that SCV-CVC are not superior in patients undergoing allogeneic, autologous HCT or ICT. Disclosure: No conflict of interest disclosed. P556

Calcineurin inhibitor-free GVHD prophylaxis with mTOR inhibitor everolimus plus mycophenolate mofetil in allogeneic-SCT for patients with hematologic malignancies: a prospective single-center, phase I/II study Schäfer H.S.1, Finke J.1, Bertz H.1, Wäsch R.1, Marks R.1 Universitätsklinikum Freiburg, Department Innere Medizin, Klinik für innere Medizin 1, Freiburg, Germany 1

Introduction: Agressive hematologic malignancies have a high relapse risk even after allogeneic stem cell transplantation (HCT). Choosing immunosupressive regimens in these patients remains challenging because GVHD prophylaxis with calcineurin inhibitors (CNI) may interfere with a possible graft versus leukemia/lymphoma effect (GVL). Everolimus, an oral mTOR-inhibitor is able to mediate antimalignancy effects and promotes the generation of regulatory T-cells. Here we report for the first time on a calcineurin inhibitor-free GVHD prophylaxis with everolimus and mycophenolate mofetil in a prospective, single-center, phase I/II study in 21 patients with AML/MDS (n = 15), ALL (n = 1), CML/MPS (n = 2) and NHL/CLL (n = 3) with unfavourable risk profile according to disease risk index (DRI). Results: All patients engrafted, 20 of 21 patients achieved complete remission of underlaying disease. 7 patients (33%) developed acute grade III-IV GVHD, 15/21 patients (71%) developed chronic GVHD (cGVHD) of all grades , 5/21 (23,8%) were severe -, 10/21 (47,6%) moderate grades respectively. No higher grade toxicity related to study medication was observed, especially no case of transplantation-associated microangiopathy (TAMA) and no acute renal failure. Median follow-up of all surviving patients is 70 month. The 1y-overall survival (OS) rate and progression free survival (PFS) rate is 76% and 67% respectively. The 4y-overall survival (OS) rate and progression free survival (PFS) rate is 48% and 43% respectively 5 Relapses occured, the cumulative risk of relaps was 24%. Conclusion: Although this new combination regimen shows relatively high rates of acute and chronic GvHD the low relapse rate is promising. In our opinion the CNI-free GVHD prophylaxis with everolimus and my-

Abstracts

cophenolate mofetil is a valid option as GVHD prophylaxis regimen in patients with considerable risk of relapse. Disclosure: No conflict of interest disclosed. P557

Ileostomy as feasible treatment option for patients with severe refractory graft versus host disease of the gastrointestinal tract after allogeneic stem cell transplantation Turki A.T.1, Basu O.2, Ditschkowski M.1, Trenschel R.1, Beelen D.W.1, Steckel N.K.1 Universitätsklinikum Essen, Westdeutsches Tumorzentrum, Klinik für Knochenmarktransplantation, Essen, Germany, 2Universitätsklinikum Essen, Klinik für Kinderheilkunde III, Essen, Germany 1

Despite numerous advances in allogeneic stem cell transplantation (aSCT) during the last decades, graft versus host disease (GvHD) remains a major source of morbidity and mortality. In particular GvHD of the gastro-intestinal (GI) tract represents a challenge for treating physicians. In refractory disease, treatment effects are often delayed or insufficient, hospitalizations extended over months and patient recovery is complicated by hospital infections. We report results of a retrospective observation performing ileostomy surgery in patients (pts) with severe refractory GI-GvHD between 2011 and 2015. Our cohort included 9 pts (n = 6 male, n = 3 female) with a median age of 48 years (range 2–53). Seven patients were transplanted because of malignant hematologic diseases, two with non-malignant diseases. 2 pts underwent a total body irradiation (TBI) based conditioning regimen and 9 pts a chemotherapy based regimen. Immunosuppressive therapy included anti-thymocyte globulin (ATG) in 4 pts 3 pts were transplanted with an identical sibling donor (ISD), and 6 pts with matched unrelated donors (MUD). Severe GI-GvHD was histologically confirmed as grade 4 in 8 pts and grade 3 in 1 patient. Prior to ileostomy all pts have all been pretreated with numerous first- and second line GvHD treatments including steroids, calcineurin- and mTOR-Inhibitors, Mycophenolat-Mofetil, ATG, Basiliximab, Infliximab, and Ruxolitinib, respectively. Interestingly, all pts presented either viral co-infections like CMV or HHV6 or bacterial co-infections. Ileostomy was performed at a median time of 58 days after development of acute GvHD. Surgery associated morbidity was low, no intervention associated mortality was observed. 5 out of 9 patients deceased during the follow up period of up to 6 years. Causes of death included other GvHD sites (n = 1 BOOP, n = 1 liver GvHD), stroke, sarcoma and cardiac arrest. In all pts a significantly decline of the feces volume could be documented after ileostomy. We also report a better control of gastrointestinal bleeding and a distinctly reduced pain level. Eight pts could be discharged at a median time of 64 days after ileostomy. In three pts ileostomy was temporary and reversal surgery was performed at a mean time of 5 months after the initial surgery. In summary we can report that Ileostomy surgery represents a feasible treatment option in severe refractory GI-GvHD. A prospective controlled randomized trial would be needed to confirm these results. Disclosure: No conflict of interest disclosed. P558

Long-term results of mTOR-inhibitor-based therapy in severe sclerodermatous chronic GvHD Jedlickova Z.1, Berg T.1, Ajib S.1, Riemann J.1, Lang F.1, Lindner S.1, Farshi P.1, Martin H.1, Serve H.1, Bug G.1 Universitätsklinikum Frankfurt, Frankfurt am Main, Germany

Introduction: Chronic graft-versus-host disease (cGvHD) is a frequent late complication of allogeneic hematopoietic stem cell transplantation. One major manifestation of cGvHD, sclerodermatous cGvHD, causes deep skin sclerosis as well as fasciitis and joint contractures. It significantly

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contributes to morbidity and mortality of long-term transplant survivors. Due to the poor response to standard immunosuppressive drugs such as calcineurin inhibitors or steroids, cGvHD remains a therapeutic challenge. Sirolimus (rapamycin) and everolimus are inhibitors of the mammalian target of rapamycin (mTOR). Besides immunosuppressive properties, they have antifibrotic and antiproliferative activity and stimulate regulatory T cells. Hence, mTOR-Inhibitors (mTOR-I) are candidates for an effective treatment of sclerodermatous cGvHD. Methods: Here we present the long-term outcome of five adult patients with severe sclerodermatous cGvHD treated with the m-TOR-I everolimus (n = 4) and sirolimus (n = 1) in our center between June 2006 and September 2015. All patients fulfilled the criteria for severe cGvHD (global scoring) and skin score 3 (organ scoring) according to the NIH-Consensus Criteria for clinical trials in cGvHD. mTOR-I were applied as first line (n = 2) or salvage therapy. As there are no approved treatment modalities besides steroids, mTOR-I were applied off-label. Drug doses were adjusted to low therapeutical levels (3–8 ng/ml). Results: At a median follow-up of 109 months (range 96–117) all patients are alive and in complete (n = 4) or partial remission (n = 1) of sclerodermatous cGvHD. Complete remission (CR) was reached at a median of 63 months of mTOR-I treatment (range 36–110). All 4 patients tapered steroids and concomitant immunosuppressants and finally stopped mTOR-I therapy after achieving CR of cGvHD. No relapse of cGvHD was observed at a median of 40 months after mTOR-I discontinuation. The main mTOR-I-related adverse events were dyslipidemia and infections. Conclusion: Our preliminary results show, that the mTOR-I sirolimus and everolimus are an effective and safe therapeutic option in cGvHD with sclerodermatous features. Our results indicate, that long-term mTOR-I treatment can lead to complete remission and withdrawal of immunosuppressants even in patients with severe sclerodermatous cGvHD. Disclosure: No conflict of interest disclosed. P559

Haploidentical second allogeneic hematopoietic stem cell transplantation for the treatment of acute leukemia relapse after first allo-HSCT: An updated retrospective registry analysis of 69 pts on behalf of the German Cooperative Transplant Group Christopeit M.1, Tischer J.2, Schetelig J.3, Bornhäuser M.3, Uharek L.4, Wolff D.5, Holler E.5, Pfrepper C.6, Behre G.6, Niederwieser D.6, Kröger N.1, Kobbe G.7, Weber T.8, Müller L.P.8, Rösler W.9, Klein S.10, Hausmann A.11, Beelen D.W.12, Bethge W.13, Schmid C.14, Kooperative Transplantstudiengruppe Universitätsklinikum Eppendorf, Klinik für Stammzelltransplantation, Hamburg, Germany, 2Universitätsklinikum München, Klinik für Innere Medizin III, München, Germany, 3Universitätsklinikum Dresden, Dresden, Germany, 4Charité-Unversitätsmedizin, Berlin, Germany, 5Universitätsklinikum Regensburg, Regensburg, Germany, 6Universitätsklinikum Leipzig, Leipzig, Germany, 7Universitätsklinikum Düsseldorf, Düsseldorf, Germany, 8 Universitätsklinikum Halle (Saale), Halle (Saale), Germany, 9Universitätsklinikum Erlangen, Erlangen, Germany, 10Universitätsklinikum Mannheim, Mannheim, Germany, 11Städtisches Klinikum München, München, Germany, 12 Universitätsklinikum Essen, Essen, Germany, 13Universitätsklinikum Tübingen, Tübingen, Germany, 14Klinikum Augsburg, Augsburg, Germany 1

Introduction: 2nd allo-HSCT frequently is the treatment of acute leukemia (AL) relapsing after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Donor change to a different yet HLA-identical donor has not resulted in significantly different outcomes when compared to choosing the same HLA-identical donor used for the first transplantation (Christopeit et al., JCO 2013). Feasibilty of using a haploidentical related donor at 2nd allo-HSCT has been shown (Tischer et al., BMT 2014). Patients and methods: We performed a retrospective analysis among 68 consecutive pts (32 female; 12 ALL) from 12 German centers to interrogate the role for 2nd haploidentical allo-HSCT for AL relapsing after 1st allo-HSCT. Median age was 39 (16–64) years. Grafts at 1st allo-HSCT were from matched related (32%), matched unrelated (35%), mismatch

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unrelated (17%), haploidentical donors (6%), and other donors, including cord blood (7%). Median duration of CR after 1st allo-HSCT was 398 (181633) days. All pts received cytoreductive chemotherapy as a treatment for relapse. CR was reached at start of conditioning for haploidentical 2nd allo-HSCT in 28%. 70% showed active disease. Disease stage was not evaluable in 3%. Myeloablative/reduced conditioning for 2nd HSCT was chosen in 12%/83%. To overcome the HLA barrier, 23 pts (33%) received grafts with ex vivo T-cell depletion (TCD), following CD3/CD19 negative or CD34 positive selection. Four pts received in vivo TCD only, 2 received no TCD, 40 pts (58%) received un-manipulated grafts utilizing high-dose cyclophosphamide post-transplant (PTCY) as backbone of GvHD prophylaxis. Results: Neutrophil engraftment was achieved after a median of 17 (8– 27) days. 52 pts (75%) achieved CR after 2nd haploidentical allo-HSCT. Out of these pts, 28 (41%) relapsed after 2nd haploidentical allo-HSCT. After a median follow-up of 270 days, 50 patents had died, 23 from leukemia, and 27 from treatment-related causes. Cumulative incidences of relapse/treatment-related death were 23 ± 5%/38 ± 6% at 1 year, and 29 ± 6%/43 ± 6% at 2 years. OS at 1/2 years from haploidentical second HSCT was 38 ± 6%/19 ± 6%. Conclusion: Haploidentical 2nd allo-HSCT is a promising approach for the treatment of AL relapse after 1st allo-HSCT. OS rates comparable to alternative treatments can be observed. Different strategies to overcome the HLA barrier seem feasible. This study was registered as NCT01997918 at clinicaltrials.gov. MC/JT and WB/CS contributed equally to this work. Disclosure: No conflict of interest disclosed. P560

Immune thrombocytopenia and hemolytic anemia after ABORhD incompatible allogeneic bone marrow transplantation in a patient with chronic myeloid leukemia (CML) Huster A.1, Nowak-Harnau S.2, Häring M.-F.1, Wirths S.1, Kanz L.1, Bethge W.1, Vogel W.1 Universitätsklinikum Tübingen, Medizinische Klinik und Poliklinik, Hämatologie und Onkologie, Tübingen, Germany, 2Zentrum für Klinische Transfusionsmedizin, gGmbH Tübingen, Tübingen, Germany 1

Immune hemolytic anemia (IHA) may occur after allogeneic bone marrow transplantation (BMT) with AB0 or Rhesus mismatches between donor and recipient. The coincidence of different immune phenomena is a rare event. A 45-year-old man diagnosed with CML in November 2012 showed no adequate treatment response to imatinib, dasatinib and nilotinib. In May 2014, he underwent an allogeneic BMT from an HLA-identical donor with red blood cell (RBC) depletion, shifting blood groups from 0 Rh negative to A Rh positive. A myeloablative regimen consisting of busulfan and cyclophosphamide was applied, while ATG, tacrolimus and methotrexate were used as immunosuppressants. Post-transplant procedures were according to standard protocols. On day 212 the patient developed an immune thrombocytopenia (ITP) coinciding with a vaccination. We started treatment with steroids and immunoglobulins (IVIGs). 300 days after transplantation anti-D antibodies could be detected for the first time. Another warm-reacting IgG antibody was also detected, which did not react with rhesus positive erythrocytes. On day 347 the patient developed an IHA (minimal Hb 4.1 g/ dl). Treatment with steroids, IVIGs, rituximab and transfusion of 34 units of packed red blood cells (PRBC) 0 Rh negative and 15 units of pooled thrombocytes (64% Rh positive) was carried out. Hemolysis was refractory to treatment, and splenectomy was performed in May 2015. Following splenectomy an increase in hemoglobin and a decline in anti-D antibodies were achieved. In August another episode of IHA occurred, which again resolved upon application of steroids and rituximab. In November 2015, hemolysis (minimal Hb 3.8 g/dl) did neither respond to steroids nor to rituximab or IVIGs. Cyclophosphamide and plasmapheresis were initi-

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ated, resulting in a reduction of hemolysis and antibody titer. In January 2016, the patient lost his complete chimerism and dasatinib was started. In February, further hemolysis occurred. The patient was treated with PRBC, steroids, cyclophosphamide, plasmapheresis and immunoadsorption. This treatment resolved hemolysis. Careful monitoring of antibody titers and maintenance therapy with cyclophosphamide is planned. Pathophysiologically, either undetected preexisting rhesus antibodies generated high titers after transplantation of RhD-incompatible hematopoiesis or autoimmune hemolytic anemia was induced due to autoantibodies with preference to D positive cells. Disclosure: No conflict of interest disclosed. P561

Non myeloablative stem cell transplantation for blast crisis chronic myeloid leukemia in elderly or medically infirm patients in the era of TKI Franke G.-N.1, Fiechtner N.1, Vucinic V.1, Jentzsch M.1, Al-Ali H.K.1, Pönisch W.1, Bill M.1, Marc A.1, Lange T.2,3, Niederwieser D.1 Universitätsklinikum Leipzig, Abt. Hämatologie und internistische Onkologie, Leipzig, Germany, 2Asklepios Klinikum Weißenfels, Weißenfelds, Germany, 3 Universität Leipzig, Medizinische Fakultät, Leipzig, Germany 1

Introduction: Non-myeloablative conditioning (NMA), made haematopoetic stem cell transplantation (HSCT) available to elderly or medical unfit patients (pts). Since the introduction of tyrosine kinase inhibitors (TKI), HSCT in chronic myeloid leukemia (CML) is reserved for patients with resistance or interolance to TKI therapy or in blast crisis (BC). Here, we evaluate the outcome of 9 consecutive pts with CML transplanted after BC at our institution. Patients: 9 pts (3 females, median age 59 years (y), range 54 - 71 y) underwent HSCT for CML between 1998 and 2014 using related (n = 2) or unrelated (n = 7) donors following a conditioning regimen consisting of 2 Gy total body irradiation and fludarabine. 5 pts had an 10/10 HLA-identical donor, while 4 pts with an unrelated donor had an antigen mismatch donor (1 antigen in 3 pts, and B- and C-antigen mismatch in 1 patient). 4 male pts had a female donor. All pts received peripheral blood stem cells. Postgrafting immunosuppression consisted of mycophenolate mofetil and cyclosporine A. All pts were in chronic phase (n = 8) or accelerated phase (n = 1) after treatment of BC. Median time from diagnosis to HSCT was 438 days (d, range 57 - 4724 d). Pretransplant treatment consisted of hydroxyurea, interferon, chemotherapy, and/or TKI. The median Gratwohl score was 6 (range 5 - 6). All pts engrafted. Results: With a median follow up of 245 d (range 12 - 4211 d), 3 pts (33,3%) are still alive and in molecular remission (MR, d4221, d1118 and d904). 1 of the 3 pts alive relapsed on day 467 and was rescued with TKI treatment and donor lymphocyte infusion. 6 pts (66,7%) developed acute GvHD grades 1–4 (grades 3–4 n = 3) and 4 developed chronic GvHD. 2 pts (22,2%) relapsed and died of their disease. 4 patients (44,4%) died due to non relapse mortality (1 myocardial infarction at d199, 1 bleeding at d512, 1 infection at d245, and 1 GvHD at d229). Due to limited numbers of pts we could not detect any parameter (type of donor, mismatch, acute GvHD 0–2 vs 3–4, chronic GvHD, Gratwohl score, stage at transplantation, age, time to transplantation) that predicts survival, although we detected a trend towards better survival with lower Gratwohl score. Conclusion: NMA-HSCT is feasible in elderly and medically unfit pts with CML in BC with overall survival of 33,3% and progression free survival of 22,2%. No parameter could be detected that associated significantly with better survival. Disclosure: No conflict of interest disclosed.

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P562

Impact of pre-transplant iron-overload on immune reconstitution and the incidence of CMV-reactivation after allogenic hematopoietic stem cell transplantation Weber T.1, Widera L.1, Ligeti K.1, Paul S.1, Müller-Tidow C.1, Müller L.P.1 University Hospital Halle, Department of Internal Medicine IV, Hematology and Oncology, Halle, Germany 1

Introduction: Several studies have shown that a general iron-overload before allogenic hematopoietic stem cell transplantation (HSCT) is associated with a higher incidence of bacterial and viral infections after transplantation. An elevated serum ferritin level has been established as a reliable and easily accessible marker for iron-overload. Only few data exist regarding the specific association between high pre-transplant ferritin levels, the rate of CMV-reactivations and its combined influence on immune reconstitution after HSCT. Methods: In a cohort of 63 patients who underwent an HSCT between January 2008 and December 2015, serum ferritin in a period of 30 days before HSCT, CMV-reactivations by quantitative PCR in peripheral blood, symptomatic CMV-infections defined by symptomatic disease clinically attributed to CMV with parallel detection of CMV-DNA, as well as immune reconstitution (subsets: CD3+CD4+, CD3+CD8+, CD20+ and CD56+ cells) until one year after HSCT were retrospectively analyzed. The patients had a median age of 55 years (range 23 to 73). The analysis included only patients with a positive pre-transplant CMV-status of either donor or recipient or both. Patients were censored in case of relapse. Results: The median pre-transplant serum ferritin was 1536 µg/l (range 11.5–5515.0 µg/l). No difference could be found among the three groups of donor-recipient-CMV-status (positive in positive, negative in positive, positive in negative; p = 0.253). A CMV-reactivation after HSCT was detected in 40 patients (63%). A symptomatic reactivation was documented for 39 patients (62%). The median pre-transplant ferritin levels in the group of patients with and without CMV-reactivation, and with and without CMV-infections were 1472 vs. 1649 µg/l (p = 0.255) and 1584 vs. 1493 µg/l (p = 0.81), respectively. In patients with serum ferritin levels below vs. above the median level, the rate of non-CMV infections till day +100 was 60% vs. 78% (p = 0.18) and the 1-year TRM rate was 34% vs. 43% (p = 0.096). No significant correlation between pre-transplant serum ferritin levels and quantity of any analyzed lymphocyte subpopulations was seen. Conclusion: This retrospective analysis shows no significant difference in the pre-transplant serum ferritin levels in patients with post-transplant CMV-reactivation compared to patients without reactivation. Thus, pre-transplant iron overload does not correlate with an increased risk of CMV-reactivation after HSCT. Disclosure: No conflict of interest disclosed. P563

Case report: a patient with acute myeloid leukemia and dyskeratosis congenita undergoing allogeneic stem cell transplantation: implications for treatment Bouillon A.-S.1, Beier F.1, Ferreira M.S.1, Kersten A.2, Küpper M.K.1, Schemionek M.1, Crysandt M.1, Silling G.1, Panse J.1, Wilop S.1, Brümmendorf T.H.1, Jost E.1 Uniklinik RWTH Aachen, Klinik für Hämatologie, Onkologie, Hämostaseologie und Stammzelltransplantation, Aachen, Germany, 2Uniklinik RWTH Aachen, Klinik für Kardiologie, Pneumologie, Angiologie und Internistische Intensivmedizin, Aachen, Germany 1

Introduction: Telomere maintenance disorders (“telomeropathies”) such as Dyskeratosis congenita (DKC) are characterized by significantly shortened telomere length (TL < 1% of age-adjusted controls), skin manifestations, bone marrow (BM) failure, liver cirrhosis and an increased risk of acute myeloid leukemia (AML). Allogeneic stem cell transplantation (aSCT) is a treatment option for BM failure in DKC but it is associated with frequent pulmonary and hepatic complications.

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Case report: A 27-year-old patient with AML was admitted to our hospital in December 2014. He had been treated with 6 cycles BEACOPP for Hodgkin´s lymphoma (HL) in 2009. On admission, the patient presented clinical signs of premature aging with hair greying and lack of fully recovered hair growth after chemotherapy (Cx) for HL. Flow-FISH analysis revealed TL below the 1% percentile within leucocytes in line with the suspected diagnosis of telomeropathy. Retrospective TL analysis by confocal Q-FISH from BM at HL diagnosis confirmed short TL. He received standard AML induction Cx (3+7), but follow-up revealed persistence of AML without any regeneration of normal hematopoiesis. Salvage Cx with FLAG-Ida was applied resulting in partial remission. The patient received an aSCT after conditioning with 140 mg/m² melphalan and fludarabin from his HLA-matched brother whose TL was found to be normal. After aSCT, he developed sepsis and sinusoidal obstructive syndrome with progressive liver failure that was treated with defibrotide and lead to ICU admission. Leucocyte count showed sufficient engraftment on day 14, however liver function recovered only partially. During critical care treatment, the patient showed cardiomyopathy, renal failure and extensive wound healing problems without epithelial proliferation indicative of severe replicative exhaustion. Finally, he died due to sepsis with acute liver failure on day 91 after aSCT. Conclusion: AML arising from DKC is a rare, possibly underdiagnosed event with substantial impact on patients´ prognosis. Therapy remains challenging due to poor BM function and high risk of organ toxicity. TL screening can help to identify patients with suspected DKC related BM failure or AML and to identify family donors without telomeropathy. Physicians should be aware of possible DKC related AML, especially in familial cases, impaired or prolonged recovery following cytoreductive treatment or coincidence of solid (e.g. oral cavity carcinomas) or hematologic malignancies. Disclosure: No conflict of interest disclosed. P564

Conditioning with thiotepa, treosulfan, fludarabin and rituximab prior allogeneic stem cell transplantation in a patient with relapsed CD20+ oligosecretory multiple myeloma Nogai A.1, Kiewe P.2, Blau O.1, Hemmati P.1, Pezzutto A.1, Arnold R.1, Vuong G.L.1, Blau I.W.1 Charité – Universitätsmedizin Berlin / Charité Centrum Tumormedizin CC14 Medizinische Klinik mit Schwerpunkt Hämatologie, Onkologie und Tumorimmunologie, Berlin, Germany, 2Medizinisches Versorgungszentrum Onkologischer Schwerpunkt am Oskar-Helene-Heim, Berlin, Germany 1

Introduction: While use of high-dose melphalan followed by autologous stem cell transplantation (HSCT) is well established, the role of allogeneic transplantation (alloTx) remains debatable. If an alloStx is indicated, the usefulness of a melphalan-based conditioning in patients after several HSCT is not clear. Therefore, establishment of other efficient regimens is needed. Patient: A 49-year old female Patient suffered from a symptomatic oligosecretory multiple myeloma with positivity of CD20 and a hyperdiploidic karyotype. Indication for therapy was a severe anemia. 3 cycles VCD, 1 cycle CAD with stem cell apheresis and HSCT with melphalan was administered. A CR was reached. 14 months after HSCT, the patient progressed with a bone marrow involvement of 70%. A therapy with lenalidomide was started, but the patient suffered from fever and abdominal pain after just one dose of lenalidomide. 2 more attempts with lenalidomide did not lead to improvement, and pomalidomide showed similar symptoms. Because of the limited therapeutic options, a second induction of bortezomib, adriamycin and dexamethason plus rituximab was started, followed by a second HSCT and a potential curative attempt with an alloStx from the sister of the patient. Since the patient had already two HSCT with melphalan, a conditioning regimen with thiotepa (5mg/kg d-6 and d-5), treosulfan (10g/m² d-8 and

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-7) and fludarabin (30mg/m² d-5 to d-2) plus rituximab (375mg/m² d-9 and d-1) was used. Leukocyte recovery was day+10, recovery of platelets day+12. A bone marrow aspiration on day +27 revealed a CR and a 100% donor chimerism. The patient suffered from a mucositis IV°, a toxic hepathopathy 1° and a GvHD 1° of the skin. There were no other severe toxicities. On day +45, nausea with emesis occurred and the patient was hospitalized. Clinically, a GvHD of the upper intestinal tract was diagnosed, treatment with budenosid lead again to fast recovery. On day+131, the patient presented with limited chronic GvHD. The multiple myeloma is still in CR. The patient has recovered from the procedure, she suffers from a limited chronic GvHD (sicca symptomatic of eyes and mouth). Discussion: We present a patient with multiple myeloma and alloStx. The conditioning regimen of thiotepa, treosulfan, fludarabine and rituximab was feasible and may be an alternative to melphalan containing regimens especially in patients having already received one or more melphalan-comtaining regimens. Disclosure: No conflict of interest disclosed.

Posterdiskussion

Lungentumoren, Kopf-Hals-Tumoren P565

An inflamed gene expression signature and immune checkpoint expression is correlated with homologous repair gene methylation in squamous cell carcinomas of head and neck, lung and cervix Rieke D.1, Ochsenreither S.1, Klinghammer K.2, Klauschen F.3, Tinhofer I.4,5, Keilholz U.1,2 Charité Comprehensive Cancer Center, Berlin, Germany, 2Charité – Universitätsmedizin Berlin, Medizinische Klinik mit Schwerpunkt Hämatologie, Onkologie und Tumorimmunologie, Berlin, Germany, 3Charité – Universitätsmedizin Berlin, Institut für Pathologie, Berlin, Germany, 4Charité – Universitätsmedizin Berlin, Klinik für Radioonkologie und Strahlentherapie, Berlin, Germany, 5DKFZ Heidelberg, DKTK partner site Berlin, Berlin, Germany 1

Introduction: Immune checkpoints are emerging treatment targets. Microsatellite instability (MSI), mutational load and checkpoint expression are imperfect predictors of treatment efficacy. We describe a link between DNA repair gene methylation and expression of immune checkpoints and an immune signature in squamous cell carcinomas. Methods: A list of DNA repair (179 genes) and immune-associated genes was established from the literature. mRNA expression and methylation status were extracted from TCGA data (cbioportal.org). Correlation analysis of DNA repair gene methylation status and expression of immune checkpoint mRNA was performed in HNSSC (279 samples) and validated in lung squamous cell carcioma, cervical carcinoma as well as other cancer types. Statistical analyses were carried out in R. Results: In HNSCC, methylation of 15 DNA repair genes was identified to be positively correlated to expression of PD-L1 and CTLA4 (cutoff: Spearman rho > 0.3). Subgroup analysis revealed statistically significant enrichment for the homologous recombination pathway (p = 0.004). Methylation of candidate genes was associated with reduced expression. Among the candidate genes, RAD51B and XRCC3 methylation showed the highest correlation with PD-L1 (r = 0.471, r = 0.472) and CTLA4 (r = 0.525, r = 0.616) expression (p values < 2.2×10-16). Methylation of these DNA repair genes was associated with the expression of other checkpoints (CD28, PD-1, TIM-3, LAG-3, VISTA, ICOS, BTLA), ligands (PD-L2, CD80/86), MHC-class 1 and T-cell associated genes (FOXP3, CD8A) as well as an interferon-gamma signature that has been shown as predictive for clinical response to checkpoint inhibitors. These findings could be reproduced in independent datasets of lung squamous and cervical carcinoma and, to various extents, also in non-squamous cancer histologies.

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Conclusion: We propose for the first time a link between DNA repair gene methylation and an inflamed gene expression signature identified in HNSCC and validated for lung and cervical cancer with similar effects also seen in other cancer types. We hypothesize that homologous recombination deficiency (HRD) may be mediated by DNA repair gene hypermethylation and linked to an immune-evasive phenotype. Correlation of DNA repair gene methylation and clinical efficacy of checkpoint inhibitors should be investigated. Disclosure: Damian Rieke: No conflict of interest disclosed. Ulrich Keilholz: Advisory Role: Bristol-Myers Squibb, Merck Serono, Astra Zeneca, MSD Oncology, Pfizer; Financing of Scientific Research: Amgen, Bristol-Myers Squibb, Merck KGaA, MSD Oncology, Astra Zeneca, Novartis; Expert Testimony: Merck Serono, Pfizer; Other Financial Relationships: Amgen, MSD Oncology, Bristol-Myers-Squibb, MSD Oncology, Novartis, Merck Serono

gression free survival of 5.0 months (both preliminary results) are comparabe to the results in the phase III EXTREME trial. Conclusion: The interim analysis of the SOCCER trial (non-selected patient cohort) is in agreement with the published efficacy and safety data of Cetuximab in combination with platinum-based chemotherapy in 1st line treatment of recurrent and/or metastatic SCCHN1. Reference: 1 Vermorken JB et al.: N Engl J Med 2008; 359: 1116–11127. Disclosure: Markus Hecht: Financing of Scientific Research: Merck; Expert Testimony: Novartis Rainer Fietkau: Advisory Role: Merck, Novartis, Pierre Fabre; Financing of Scientific Research: Merck, BrainLab, Fresenius, Novartis, Roche, Pierre Fabre P567

P566

Cetuximab in combination with platinum-based chemotherapy or radiotherapy in recurent and/or metastatic SCCHN in a non-selected patient cohort (interim analysis of the phase IV SOCCER trial) Hecht M.1, Hahn D.2, Beutner D.3, Reichert D.4, Göhler T.5, Wurm R.6, Welslau M.7, Renziehausen L.8, Balermpas P.9, Bergmann T.10, Aßmann M.11, Belka C.12, Orlowski K.13, Finzsch M.13, Illerhaus G.2, Fietkau R.1 Universitätsklinikum Erlangen, Friedrich-Alexander-Universität ErlangenNürnberg, Strahlenklinik, Erlangen, Germany, 2Katharinenhospital Stuttgart, Hämatologie, Onkologie und Palliativmedizin, Stuttgart, Germany, 3University of Cologne, Department of Otorhinolaryngology, Head and Neck Surgery and Center for integrated Oncology Köln Bonn, Köln, Germany, 4Onkologie Westerstede Aurich Rhauderfehn, Medizinische Studiengesellschaft NordWest GmbH, Westerstede, Germany, 5Onkozentrum Dresden/Freiberg, Dresden, Germany, 6Klinikum Frankfurt (Oder), Klinik für Strahlentherapie und Radioonkologie, Frankfurt (Oder), Germany, 7Klinikum Aschaffenburg, Hämato-Onkologische Schwerpunktpraxis, Aschaffenburg, Germany, 8 Onkologische Schwerpunktpraxis Drs. Renziehausen, Chemnitz, Germany, 9 Universitätsklinikum Frankfurt, Goethe-Universität Frankfurt, Klinik für Strahlentherapie und Onkologie, Frankfurt (Main), Germany, 10SRH WaldKlinikum Gera, II. Medizinische Klinik, Gera, Germany, 11MVZ Elblandpolikliniken, Praxis Dr. Aßmann, Riesa, Germany, 12Klinikum der Universität München, Strahlentherapie und Radioonkologie, München, Germany, 13Merck Serono GmbH, Darmstadt, Germany 1

Background: Cetuximab in combination with platinum-based chemotherapy followed by cetuximab monotherapy until progression significantly prolonged overall survival (OS) in patients with recurrent and/ or metastatic squamous-cell carcinoma of the head and neck (SCCHN) (EXTREME trial)1. SOCCER is a prospective, non-interventional study to evaluate symptom control in patients with recurrent and/or metastatic SCCHN treated with Cetuximab in combination with platinum-based chemotherapy or radiotherapy. Methods: 219 patients with recurrent and/or metastatic SCCHN were enrolled. Full information on the concomitant therapy were available for 196 patients. In the preplanned interim analysis response rates for the different Cetuximab containing therapy regimes as well as reasons for treatment discontinuations were evaluated. Results: 82 and 76 patients received Cisplatin+Cetuximab or Carboplatin+Cetuximab, respectively. 38 patients were treated with radiotherapy+Cetuximab for recurrent disease. Compared to the EXTREME trial, this study included more patients with metastatic disease (62% vs 47%) and more patients with an ECOG score of ≥2 (23% vs 12%). Median treatment duration of Cetuximab was 4.8 weeks in combination with radiotherapy and 11 weeks in combination with chemotherapy. Main reasons for Cetuximab treatment discontinuation was tumor progression (44%), death (14%) and patients wish (12%). Treatment was discontinued due to toxicities in 7% of the patients. Response data were available in 103 patients. The overall response rate (ORR) and the disease conrol rate (DCR) were 50 / 70% in the radiotherapy group, 43 / 76% in the Cisplatin group and 31 / 71% in the Carboplatin group, respectively. The median OS of 8.8 months and the median pro-

Abstracts

Nivolumab induced an inflammatory swelling of submandibular glands in a patient with metastatic squamous non-small cell lung cancer Frille A.1, Oltmanns A.1, Wirtz H.1 Universitätsklinikum Leipzig, Pneumologie, Leipzig, Germany

Introduction: The immune-checkpoint inhibitor nivolumab, a fully human immunoglobulin G4 monoclonal antibody against programmed death 1 (PD-1) receptor, received European approval for patients with recurrent advanced or metastatic squamous non-small cell lung cancer (sqNSCLC) after initial platinum-based chemotherapy in July 2015. Immune-related adverse events (IrAEs) are associated with the targeted release of PD-1 pathway-mediated inhibition of anti-tumor immune response due to uncontrolled enhancement of T cell function. Case: A 57-year-old male Caucasian was administered nivolumab (3 mg/ kg) due to progression of metastatic sqNSCLC after six courses of cisplatin/vinorelbine. Within 13 days, the patient presented with a visible, progressively growing painless but inflamed swelling of the anterior cervical region. He denied shortness of breath, dysphagia, fever or night sweats. Clinical examination did not show a palpable stone at the sublingual fold or caruncle and was unsuspicious for the remaining organ systems. Immediate cervical ultrasound revealed a bilateral swelling of the submandibular glands (28 mm in diameter) without evidence of abscess formation, dilatation of submandibular duct or associated pathologic cervical lymph nodes. Since no common cause of submandibular gland swelling was found, we assumed a nivolumab-induced immunoreaction to the submandibular glands. We started an oral regimen of prednisolone (1 mg/kg daily) and cefuroxime (250 mg bid) for seven days. Prednisolone was subsequently tapered the following 7 days. Ultrasound re-examination on day 9 of prednisolone therapy showed decrease of swelling of both submandibular glands (23 mm in diameter). Nivolumab was reinitiated 28 days after first administration without any further similar irAEs ever since. Conclusion: Here, we report an uncommon nivolumab-mediated inflammatory swelling of the submandibular glands. IrAEs due to PD-1 inhibitor nivolumab may affect any organ system. Typically, skin (rash, mucosal irritation), gastrointestinal (diarrhea, colitis, hepatitis), and endocrine systems (hypophysitis, hypothyroidism with nausea, headache, and fatigue) are affected. Less commonly, pneumonitis, nephritis, pancreatitis, neurologic and hematologic syndromes may occur. Treatment recommendations consist of discontinuation of nivolumab and administration of corticosteroids until signs and symptoms are sufficiently resolved. Disclosure: No conflict of interest disclosed.

Oncol Res Treat 2016;39(suppl 3):1–192

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P568

Targeting HER2- and MET co-amplification in patients with EGFR mutation resistant to EGFR-TKI therapy Brandes V.1, Scheffler M.1, Kambartel K.2, Ortiz-Cuaran S.3, Scheel A.4, Michels S.5, Nogova L.5, Fischer R.5, Abdulla D.5, Merkelbach-Bruse S.4, Büttner R.4, Sos M.L.3, Wolf J.5, Lung Cancer Group Cologne Uniklinik Köln, Klinik I für Innere Medizin, Lung Cancer Group Cologne, Köln, Germany, 2Krankenhaus Bethanien, Moers, Germany, 3Uniklinik Köln, Department of Translational Genomics, Köln, Germany, 4Uniklinik Köln, Institut für Pathologie, Köln, Germany, 5Uniklinik Köln, Klinik I für Innere Medizin, Köln, Germany 1

Purpose: Both HER2 amplification and MET amplification have been described as modes of resistance to either first-generation EGFR-TKI therapy with and without EGFR T790M mutation or third-generation EGFR-TKI therapy. While both aberrations act as potential targets for subsequent therapies, i. e. afatinib plus cetuximab in HER2-cases and the addition of crizotinib for MET-amplified patients, little is known about a potential co-amplification of both HER2 and MET. We set out this study to identify patients with co-amplification and EGFR mutation and to analyze its impact on treatment and course of the disease. Finally, we propose preliminary treatment recommendation. Patients and methods: Three patients with EGFR mutation were detected so far with co-amplification of HER2 and MET, identified using fluorescence in-situ hybridization (FISH). In all cases, next-generation sequencing (NGS) was performed in order to detect concomitant driver aberrations. Functional assays to prove the findings are now ongoing. Results: One female patient with EGFR del19 became finally resistant to erlotinib, after a 6-year history of erlotinib and intermittent “cyberknife” radiation. In the absence of an EGFR T790M mutation, concomitant HER2 amplification and low-level MET amplification (present from the initial diagnosis) were found. We started treatment with afatinib monotherapy, which resulted in primary progression. The switch to afatinib plus cetuximab also led to PD, whereas the combination of afatinib and crizotinib led to an impressive partial response. A second patient with EGFR mutation, this time L858R, presented with progression after again 6 years of treatment history with erlotinib. Both amplifications could be found, beside an NRAS mutation and without T790M. The proposed combination with afatinib and crizotinib led do an impressive PR, too. A third male patient became progressive under treatment with AZD9291, having both amplifications beside the EGFR del19 and EGFR T790M mutation. So far, he is treated locally and has SD. Conclusion: Additional functional and patient-derived data is now collected. We propose that the combination of afatinib and crizotinib has promising activity in patients without T790M. Further examination will provide insights into the treatment options of patients progressing with these amplifications under third-generation EGFR TKIs with the presence of T790M. Disclosure: No conflict of interest disclosed. P569

EATON: An open-label, multicenter, phase I dose-escalation trial of EGF816 and trametinib in patients with non-small cell lung cancer and acquired EGFR p.T790M positive resistance to 1st or 2nd generation EGFR TKI therapy – a trial in preparation Michels S.1, Brandes V.1, Thurat M.1, Nogova L.1, Scheffler M.1, Fischer R.1, Merkelbach-Bruse S.2, Hellmich M.3, Büttner R.2, Wolf J.1, Lung Cancer Group Cologne Uniklinik Köln, Klinik I für Innere Medizin, Lung Cancer Group Cologne, Köln, Germany, 2Uniklinik Köln, Institut für Pathologie, Lung Cancer Group Cologne, Köln, Germany, 3Universität zu Köln, Institut für Medizinische Statistik, Informatik und Epidemiologie (IMSIE), Köln, Germany 1

Introduction: Multiple mechanisms of resistance to 1st and 2nd generation EGFR TKI therapy in EGFR mutated NSCLC have been described, most often including the acquisition of the gatekeeper mutation EGFR

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p.T790M. Preclinical models and clinical findings have also shown that co-occurring activation of the RAS/MEK pathway may result in reduced EGFR dependency and resistance to 3rd generation EGFR TKI therapy. Co-inhibition of MEK restores sensitivity in pre-clinical models. We thus hypothesize that the combined inhibition of EGFR and MEK may be effective in patients with acquired RAS/MEK activation and may as well prevent the acquisition of RAS/MEK mediated resistance to 3rd generation EGFR TKIs. Methods: EATON is an international, multicenter, phase I, dose escalation, investigator initiated trial investigating the recommended phase 2 dose (RP2D), safety and preliminary efficacy of the combination of the 3rd generation EGFR inhibitor EGF816 with the MEK inhibitor trametinib. Patients with advanced non-small cell lung cancer harbouring sensitizing EGFR mutations (EGFR del19 or EGFR p.L858R), with progression upon treatment with 1st or 2nd generation EGFR TKI and acquired resistance mutation EGFR p.T790M will be eligible for the trial providing high level amplification of MET has not been detected. Dose level escalation will be based on a modified traditional cumulative 3+3 design, i.e. “up and down” (dose level 1: 100 mg EGF816 QD + 1 mg trametinib QD). A total number of 24 patients will be enrolled in 7 trial sites in Germany and Spain. At a first stage, 18 (6×3) patients will be treated and evaluated. Based on this data, the “virtual MTD” is estimated and 6 further patients (2×3) will be treated on a dose level equal or below the virtual MTD. Exploratory endpoints aim at the identification of potential mechanisms of resistance to the trial treatment by massively parallel sequencing (MPS), FISH, phospho-protein analyses and whole exome/genome sequencing of baseline and PD biopsy tumour tissue. Additionally, blood samples for MPS of cell free DNA will be collected throughout the trial treatment. Results and conclusions: Since this is a trial in preparation, no formal results will be presented. The initiation of the trial is planned for Q3/2016. An interim analysis will be made after full recruitment of the trial for determination of the RP2D and the safety profile. Disclosure: Sebastian Michels: Advisory Role: Novartis, Pfizer, Boeringer Ingelheim; Financing of Scientific Research: Novartis, Pfizer, Boeringer Ingelheim Jürgen Wolf: Advisory Role: Novartis, Pfizer, Boeringer Ingelheim, Astra Zeneca; Financing of Scientific Research: Novartis, Pfizer, Boeringer Ingelheim, Astra Zeneca P570

Individualized sequential therapy in patients with metastatic lung cancer and actionable oncogenic alterations: two case reports of prolonged survival Peters K.1, Toenges R.2, Weiß S.2, Förster S.3, Krammer-Steiner B.1 Klinikum Südstadt Rostock, Klinik für Innere Medizin III, Rostock, Germany, Diagenom GmbH, Rostock, Germany, 3Klinikum Südstadt Rostock, Klinik für Allgemein-, Viszeral-, Thorax- und Gefäßchirurgie, Rostock, Germany 1 2

Introduction: Despite progress in the therapy of advanced lung cancer, mortality remains high. The discovery of actionable EGFR mutations and ALK translocations have identified subsets of patients with predictable tumor response to oral targeted drugs. Patients and methods: We present two advanced NSCLC patients with extended survival while undergoing targeted therapy according to the presence of oncogenic alterations. The first patient (female, 65 years old, diagnosis stage IV NSCLC 12/2011) demonstrated an EGFR deletion of exon 19 in primary tumor tissue. Initial therapy consisted of gefitinib (12/2011 to 06/2013). Brain metastases required whole brain irradiation therapy (03–04/2013). During disease progress she received four cycles of intravenous cisplatin/alimta chemotherapy followed by maintenance therapy with alimta until 04/2014. During further disease progress, therapy was changed to oral afatinib (from 04/2014). Increasing tumor marker concentrations indicate further tumor progress. EGFR T790M analysis is in work to evaluate further TKI treatment options. The second patient (male, 53 years old, diagnosis stage IV NSCLC 05/2013) demonstrated an ALK rearrangement in primary tumor tissue. Initial therapy consisted of intravenous cisplatin/vinorelbine accompanied by grade IV hematolog-

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ic and renal toxicity after two cycles and followed by crizotinib therapy (09/2013 to 03/2015). His individual clinical response allowed for atypical partial lung resection after PET imaging. Resected tumor tissue was again positive for the ALK rearrangement. Brain metastases required whole brain irradiation therapy (02/2015). During disease progress, ceritinib therapy was started as individualized treatment decision (prior to drug approval). Mucositis grade IV developed after two weeks of treatment and therapy was halted until 02/2016. PET imaging indicated tumor progress and crizotinib was again started followed by reduced dose ceritinib. Conclusions: Two advanced stage NSCLC patients with the presence of actionable oncogenic alterations in primary tumor tissue have been undergoing sequential therapy resulting in survival of 4.5 and 3 years, respectively. These cases demonstrate that in addition to guidelines and recommendations, individualized sequential treatment contributes to improving survival rates in advanced stage NSCLC patients. Disclosure: No conflict of interest disclosed. P571

Case report: Long-lasting remission (9+ months) was seen in a NSCLC adenocarcinoma patient with Nivolumab after 4 years of several chemotherapy regimen and TKI treatment Fröhling K.-P.1, Neumeister W.1 Kath.Klinikum Koblenz- Montabaur, Pneumologie, Koblenz, Germany

Introduction: Nivolumab is a human Immunglobuline-G4 (IgG4) monoclonal antibody binding to the Programmed Death-1-(PD-1) receptor which blocks the interaction of the receptor with the ligands PD-L1 and PD-L1. Nivolumab enhances the T-cell reactions including tumor.It is approved in July 2015 for the treatment of squamous histology NSCLC and in April 2016 for non-squamous NSCLC. Methods: We report here a case of a 73-year-old female with multiple pretreatments with adenocarcinoma of the lung. First diagnosis in 2010: right lower lobe PT2, pN0 , M0 G2.A lower lobe resection was performed followed by 2 cycles adjuvant chemotherapy (Paclitaxel +Cisplatin). A relapse was documented in July 2011 pulmonary mets) Cemotherapy was started and a stabilization of the disease with some periods of tumor response for 3 years. The Patient on almost continuous treatment (07.11–05.15)1st line: Pemetrexed (07.11- 03.12) best Response SD, 2nd line: Gemcitabine (03.12- 08.12) best Response PR, 3rd line: Erlotinib (12.12- 03.13) PD, 4th line Pemetrexed (05.13- 01.2014) PR, 5th line Navelbine oral 01.14–07.14 best Response SD, 6th line: 07.14 Docetaxel+Carboplatin best Response SD, stopped due to intolerance.After these 2 years a „Drug-Holiday“of 3 months was realized. Due to the previous responses to Pemetrexed we have been retreating the patient with Pem for 2 months starting in 09.14 leading to a Stable disease/minor progress only. After ongoing therapy unfortunately a progress was seen after 2 more months. Due to negative EGFR, MET or and ALK mutation status at this time point we initiated Docetaxel+Nintedanib in 03.15 and were not able to stop progress after 2 months. We started a therapy with Nivolumab in July 2015. At that time, off-label use as an individual therapeutic trial. After 3 months we saw a PR lasting 9 months. Please refer to the documented CT images. (remission in primary lung tumor and lung mets) The patients is in stable general condition. The therapy was well tolerated with mild GI toxicities (diarrhea grade 1–2). She is still on Nivolumab treatment (April 16). Conclusion: After several chemotherapy regimen and one TKI treatment approach (with short PR or SD benefits) we have seen with Nivolumab treatment a long-lasting remission of 9+ months in a 73-year old female patient. She has a good general condition and mild GI toxicities. She is on further Treatment (April 2015).

P572

RAPTOR, the global phase II, III clinical development program of the PD-L1 inhibitor Durvalumab (MEDI4736) in 2nd-line recurrent/metastatic (R/M) squamous cell carcinoma of the head and neck (SCCHN) - A German update on the HAWK, EAGLE and CONDOR trials Rüssel J.1, von Einem J.2, Keilholz U.3, Dietz A.4, Grünewald V.5, Maschmeyer G.6, Kasper S.7, Rupprecht M.8, Sasse B.8, Melillo G.9, Krauss J.10 University Hospital Halle, Halle/Saale, Germany, 2Ludwig-Maximilian-University Hospital Munich, Munich, Germany, 3Charité Berlin, Berlin, Germany, 4University Hospital Leipzig, Leipzig, Germany, 5Hannover Medical School, Hannover, Germany, 6Ernst-von-Bergmann Clinic, Potsdam, Germany, 7West German Cancer Center Essen, Essen, Germany, 8AstraZeneca GmbH, Wedel, Germany, 9 AstraZeneca PLC, Gaithersburg, United States, 10National Center for Tumor Diseases Heidelberg, Heidelberg, Germany 1

Introduction: SCCHN is the sixth most common cancer in the world as well as in Germany, with more than 500.000 and 15.000 newly diagnosed patients (pts) each year, respectively. The prognosis for R/M SCCHN is poor. Approved standard treatment is a platinum-based therapy (PLT) plus the anti-EGFR monoclonal antibody Cetuximab, providing a median overall survival (OS) of 10.1 months. Currently, there are no standard options used after failure of 1st line treatment. A significant number of SCCHNs are characterized by a highly immunosuppressive tumor microenvironment resulting in functional loss of activity of immune effector cells. Monoclonal antibodies (mAb) targeting inhibitory immune receptor interactions of T cells such as the PD-L1/PD-1 or B7/CTLA-4 axis have been shown to very effectively restore T cell immunity in several cancers including SCCHN. Durvalumab is a selective human IgG1 mAb that blocks PD-L1 binding to PD-1 and CD80, whereas Tremelimumab is a selective human IgG2 mAb inhibitor of CTLA-4. Durvalumab monotherapy has shown promising antitumor activity and a manageable safety profile in SCCHN in a Phase I/II study (NCT01693562) as well as in combination with Tremelimumab in a Phase Ib study in pts with NSCLC (NCT02000947). Methods: RAPTOR is a comprehensive global late phase clinical development program in 2nd line R/M SCCHN. The HAWK, CONDOR and EAGLE trials will enroll pts after failure of PLT for R/M with a PD-L1+, PD-L1- and PD-L1+/- SCCHN, respectively. Primary Endpoints of the phase II trials HAWK and CONDOR are ORR (RECIST v1.1) and OS for the phase III EAGLE study. Secondary Endpoints contain DCR, DoR, PFS, OS, Safety (CTCAE v4.03)/Tolerability & HRQoL. See table for exploratory endpoints.

Disclosure: No conflict of interest disclosed.

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–192

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Tab. 1 to P572. RAPTOR-Trials

Study

Phase N

Randomi- PD-L1-status Stratification zation (Cutoff: 25%) Factors

HAWK

112

n/a

PD-L1+

–

CONDOR

240

2:1:1

PD-L1-

HPV, smoking history

EAGLE

III

720

1:1:1

PD-L1+ PD-L1-

HPV, smoking history

Study-design Durvalumab (10 mg/kg IV q2w up to 12 mo.) monotherapy 1. Durvalumab (20 mg/kg IV) + Tremelimumab (1 mg/ kg IV) combination 2. Durvalumab monotherapy (10 mg/kg IV) 3. Tremelimumab monotherapy (10 mg/kg IV) 1. Durvalumab (10 mg/kg IV up to 12 months) monotherapy 2. Tremelimumab (1 mg/kg IV) plus Durvalumab (20 mg/kg IV for up to 12 months) combination 3. SoC (Cetuximab, Taxane, Methotrexate, or Fluoropyrimidine)

Results: To date, the participating German centres have enrolled/randomized 11/20/4 pts and 6/12/2 pts in HAWK, CONDOR, and EAGLE, respectively. Preliminary data confirm the manageable safety profile known from the previous trials. The trials are accompanied by translational research. Conclusions: Patients with R/M SCCHN have a poor overall prognosis, and there is an unmet need for new treatment options. The RAPTOR trials will determine the safety and efficacy of Durvalumab, +/- Tremelimumab, in this difficult-to-treat patient population. Disclosure: No conflict of interest disclosed. P573

Oral cavity carcinomas as primary manifestation of telomere maintenance disorders (telomeropathies) Beier F.1, Schmitt K.1, Bouillon A.-S.1, Hasenbank C.1, Vankann L.1, Ambrosch P.2, Hoffmann T.3, Fietkau R.4, Panse J.1, Brümmendorf T.H.1 Klinik für Hämatologie, Onkologie, Hämostaseologie und SZT, Med. Fakultät Uniklinik RWTH Aachen, Aachen, Germany, 2Klinik für Hals-NasenOhrenheilkunde, Universitätsklinikum Kiel, Kiel, Germany, 3Klinik für HalsNasen-Ohrenheilkunde, Universitätsklinikum Ulm, Ulm, Germany, 4Klinik für Strahlentherapie, Universitätsklinikum Erlangen, Erlangen, Germany 1

Introduction: Telomere maintenance disorders (“telomeropathies”) such as Dyskeratosis congenita (DKC) are characterized by mutations in the telomerase complex resulting in significantly shortened telomere length (TL) below the 1% percentile of age-adjusted controls. Bone marrow (BM) failure or skin manifestations are frequently the first signs of DKC in young adults. Radiation or cytoreductive treatment of DKC patients is associated with high toxicity and organ failure due to the impaired cellular regenerative capacity. Retrospective studies showed that DKC patients have an approx. 1000-fold increased risk for development of oral cavity carcinomas (OCC). However, no data are available about OCC as first manifestation of DKC. In this study, we performed TL screening of atypical OCC patients (< 50 years) without apparent risk factor (HPV negative, non-smokers) for possible telomeropathies. Patients and methods: TL screening was carried out using flow-FISH in patients with OCC from three different centers. All patients were included into the “Aachen Telomeropathy Registry”. Mean age of the first 10 patients presented in this interim analysis was 42.2 years. Results: TL screening revealed TL below the 1% percentile in the granulocytes and lymphocytes in one out of 10 patients while nine patients showed TL within normal range. Mutation analysis of genes from the telomerase complex of the patient with TL below the 1% revealed mutation in the TERC gene (r.50C>A).

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Endpoints

Clinical Trials. gov Identifier

1: ORR 2: DCR, DoR, PFS, OS & HRQoL

NCT02207530

1: ORR 2: DCR, DoR, PFS, OS , Safety/tolerability & HRQoL

NCT02319044

1: OS 2: ORR, PFS, DCR, DoR, proportion of patients alive and progression free at 6 & 12 months, Safety/Tolerability, HRQoL Expl. NCT02369874 PK, immunogenicity, potential biomarkers that may influence the progression of cancer and/ or prospectively identify patients likely to respond to treatment.

Conclusion: OCC as first manifestation of DKC is a rare incident. Nevertheless, we suspect significant underdiagnoses of DKC in patients with atypical presentation (i.e. young age, non-smokers, no excess alcohol consumption, HPV-negativity). In our highly selected, mixed prospective and retrospective cohort of “atypical” OCC patients, we found one out of 10 patients with a degree of TL shortening indicative of a telomeropathy. As cytotoxic treatment of OCC can be associated with excess toxicity in patients with telomeropathies, physicians should be aware of a possible DKC despite the rarity of the condition. This holds true especially in cases with concurrent family history of hematologic malignancies (acute myeloid leukemia or BM failure) or associated liver or pulmonary fibrosis. Our ongoing German multicenter investigation aims to establish TL screening to identify patients with a telomeropathy background in atypical OCC. Disclosure: No conflict of interest disclosed. P574

Nivolumab (Optivo®) in squamous cell carcinomas (SCC) in head and neck cancer – a case report Severin K.1, Stupp C.2, Schmitz S.1, Steinmetz T.1 Practice, Sachsenring Cologne, Hematologic and Oncologic Practice, Cologne, Germany, 2St Elisabeth Hospital Cologne, Department of ENT (Otolaryngology), Cologne, Germany 1

Introduction: The immuno-therapeutic drug nivolumab (optivo®) is nowadays widely accepted as a highly potent approach in therapy of squamous cell carcinomas of the lung. However little data is available on feasibility and efficacy in (SCC) in head and neck cancer Methods and Results: Case report of a 52 year old patient first diagnosed in 03/06 with (SCC) of the left tonsil, radical left side laser-tonsilectomy/ conventional right side tonsillectomy, modified radical neckdissection level III: G2, L1, HPV positive, pT2 (>2 cm), pN2b(3/15), M0, IVA postoperative radio-CTX(platinum/5FU). First relapse in 01/07, laser-surgical resection left lateral pharynx and palate, R1, neck exploration, additive radiotherapy. Second relapse in 12/2007, locally velum, palatal resection with plastic coverage, R0, PET/ CT scan 08/08 with no active malignancy, thus maintenance therapy with cetuximab mono weekly for 2.5 years. 03/15 after 5 year CR, local recurrence oropharynx, resection, pectoralisflap: rpT3, G2 08/15 local recurrence in oropharynx. In MRI suspicious infiltration in atlas, dens-axis, left lateral tongue, left hypopharynx. No option for surgery (decision ENT department), nor radiation (decision radiotherapist as 100 Gy total dose). Therefore 08/15–12/15: 16 cycles of a metronomic therapy weekly low-dose nabpaclitaxel(150 mg absolute dose)/carbopla-

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tin(100 mg absolute dose), no side effects other than mild alopecia but good response (regression up to 1/5 of the original tumor extent in oropharynx li pharyngeal wall. CT scan thorax/abdomen 11/15 showing no distant metastases. 01/16 local progression with infiltration into the deep neck muscles, thus 3 rd line palliative CTX with vinorelbine mono (50 mg/m2 d1,8,15 qd29) with SD 03/16 local progression in left oropharynx resulitng in initiation of (off-label use) immunotherapy with nivolumab (optivo®) as patient is still in excellent condition and tumor turned out to be highly positive for PDL1 (assumed effectiveness of checkpoint modulation). After 3 weeks local progression (no side-effects), continuation of therapy, currently after 4thcycle (week 8) considerable regression with merely slight ulceration. We will carry on treatment, clinical course has to be followed. Conclusion: Nivolumab (Optivo®) showed to be feasible and seems to be highly efficient in (SCC) in head and neck cancer. Although further investigation is needed to evaluate influence of HPV positivity and manifestation of PD-L1 on outcome Disclosure: No conflict of interest disclosed. P575

Treatment with checkpoint-inhibitor immunotherapy is associated with an increased rate of hypothyreodism: A single center evaluation of Pembrolizumab treatment in fifteen patients with metastatic malignant melanoma Stubbe F.1, Große-Thie C.1, Schäd-Trcka S.2, Freitag S.1, Timmer K.1, Lakner J.1, Henze L.1, Emmert S.2, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany, 2Department of Dermatology, Rostock University Medical Center, Rostock, Germany 1

Introduction: The introduction of checkpoint inhibitors led to an enormous progress in the immunotherapeutic treatment of advanced metastatic malignant melanoma. Pembrolizumab represents a PD1 antibody which was approved for malignant melanoma treatment in Germany in 2015. Herein, we report a first evaluation of Pembrolizumab treatment efficiency and side effects in malignant melanoma patients treated at our center. Patients and methods: Since July 2014 fifteen patients (8 females (53%) and 7 male (47%)) with advanced metastatic malignant melanoma were treated. All patients received the PD1 antibody Pembrolizumab (2 mg/ m²BSA, q 3 weeks) in our center. Responses to the treatment were evaluated regularly by clinical examinations, blood tests and CT, PET-CT or MRI. Results: One to 29 courses (median 11 courses) were administered. At the time of evaluation in April 2016 one complete remission was achieved (7%) while 2 patients (13%) achieved stable disease. A mixed response was observed in 5 patients (33%) and progressive disease in 7 patients (47%). Five patients died (33%) due to disease progression. The main observed adverse event was treatment-requiring hypothyreodism in 5 patients (33%). Hormon substitution therapy was initiated without interrupting the PD-1 therapy. Conclusion: The PD1 antibody Pembrolizumab resulted in objective response and disease stabilization in 53% of all patients. Hypothyreodism was more often observed than in the clinical trials performed. Disclosure: No conflict of interest disclosed.

P576

Clinical characteristics of HIV-associated Kaposi’s sarcoma in the era of combination antiretroviral therapy Meyer L.1, Stoehr A.2, Horst H.-A.1, Plettenberg A.2, Lorenzen T.2, Schewe K.3, Sabranski M.3, Hoffmann C.1,3 UKSH, Kiel, Germany, 2IFI Institute, Hamburg, Germany, 3ICH Study Center, Hamburg, Germany 1

Introduction: Although a substantial reduction in the incidence of Kaposi’s sarcoma (KS) has been seen with the introduction of combination antiretroviral therapy (cART), KS remains the most common malignancy in patients with HIV infection. The progression is very variable and the optimal therapy is undefined. Methods: Retrospective analysis of all cases of KS occurring in HIV-infected patients from 2000 to 2015 in three participating German centers. Clinical characteristics, staging (tumor-related criteria by AIDS Clinical Trial group), treatment, outcome and predictive factors were evaluated. Results: During the observation period, 230 patients (226 males, 4 females) were diagnosed with HIV-associated KS. The median age at KS diagnosis was 40 years (range, 21–75 years). Median absolute and relative CD4 T-cells were 155/µl (range, 1–1069/µl) and 11% (range, 0–57%), respectively. A profound immune deficiency of less than 100 CD4 T-cells/µl was found in 35.7% while 20.0% had only limited immune deficiency with more than 350 CD4 T-cells/µl. Median HIV RNA was 5.01 log copies/ml (range, < 1.20-6.72 log copies/ml). In total, 22/230 patients (9.6%) developed KS while on current cART, among them only 9 (3.9%) with an HIVRNA of less than 50 copies/ml (median duration of virus suppression 8 months). In all cases (n = 3) with virus suppression of > 12 months at KS diagnosis, manifestations resembled those of classic KS with stable and localized cutaneous lesions at the lower extremities. After a median follow up of 5.57 years, 39 patients have died (total mortality 16.96%). Main causes of death were other AIDS defining illnesses (n = 20), other cancers (n = 5) and cardiovascular events (n = 3). In only 5 patients, KS was considered to be among the main causes of death. Low CD4 T-cells and prior AIDS but not clinical KS stage were associated with poorer outcome. Conclusions: The vast majority of HIV-associated KS cases occurs in patients without antiretroviral therapy and without viral suppression. In the setting of a long lasting virus suppression KS is very rare. The high proportion of antiretrovirally untreated KS patients with severe immune deficiency suggests missed opportunities for an earlier initiation of cART. However, disease-related mortality of KS is low and prognosis of KS patients appears to be mainly determined by HIV-related factors such as the degree of immune deficiency. Disclosure: No conflict of interest disclosed.

Posterdiskussion

Palliativmedizin, Integrative Onkologie P577

Fulminant improvement in a patient with prostate cancerassociated-TTP (CA-TTP) after initiation of enzalutamide therapy in a Palliative Care Unit (PCU) Schuler U.S.1, Linné C.2, Kroschinsky F.3, Link C.4, Heller S.5 Universitätsklinikum Carl Gustav Carus, Medizinische Klinik I und PalliativCentrum, Dresden, Germany, 2Niedergelassener Urologe, Dresden, Germany, 3Universitätsklinikum Carl Gustav Carus, Dresden, Germany, 4 Universitätsklinikum Carl Gustav Carus, Medizinische Klinik I, Dresden, Germany, 5Universitätsklinikum Carl Gustav Carus, PalliativCentrum, Dresden, Germany 1

Introduction: CA-TTP is a rare syndrome with a frequently dismal prognosis. A recent review discussed only 168 documented cases in the world literature [1]. A patient with prostate cancer and a fulminant improvement of CA-TTP during enzalutamide therapy is presented.

Abstracts

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Case report: 80 yr old patient admitted to PCU with hematoma of 30– 40% of BSA, rectal bleeding, ECCOG 3–4 and cachexia. Metastatic prostate cancer for > 8yrs. THR-penia with 26 Gpt/l, Quick 38% (INR 2,03) and hypofibrinogenemia of 0,56 g/dl. Treatment with plasma, tranexamic acid and transfusion of 1U of THR had been given on an outpatient basis. Results: Fragmentocytes of 22/1000 and LDH of 14 µmol/(s*l) [ULN 3,75] confirmed CA-TTP. Haptoglobin and hemopexin were in the lower range of normal. Referral to PCU was based on the assumption of a demise of the patient within few days. After consultation with his urologist and informed consent enzalutamide was initiated despite only limited hope of success.However, prompt clinical and laboratory improvement was observed. Within 5d fibrinogen went from life-threatening-low to elevated levels and to THR >100 Gpt/l. Tranexamic acid was discontinued and anticoagulation was initiated. Due to the patient’s general condition, specialized palliative home care was initiated but had to be discontinued upon further improvement. The prompt remission of TTP was associated with a much slower and moderate improvement of PSA-levels. Coagulation profile was still normal after one year except for a moderate elevation of d-dimers. In the meantime manifestations of Parkinson’s Disease moved to the forefront of the patient’s condition. Conclusions: This patient’s course is remarkable in several ways: (a) CR-TTP carries a dismal prognosis, successful therapy is only possible occasionally [1], and more related to tumor-directed measures than to plasma administration or plasma exchange. (b) The degree and speed of recovery was regarded as absolutely unusual. Additionally it is of interest, that this effect on the coagulopathy was seen with only modest improvement of PSA. (c) According to current recommendations tumor directed therapies close to the patient’s death are registered as indicators of poor quality of care. For this patient we had reason to believe, that this criterium could be fulfilled. One has to keep in mind that this can negatively affect meaningful treatment decisions.

cepts. This setting allows for the identification of patients with advanced diseases and high needs for palliative care, qualifying for palliative complex treatment. In extension it enables to look at the symptom burden of patients and relatives not yet being too advanced and without unfavorable prognosis. On the basis of individual situations short inputs on palliative thoughts, communication aids and newest research can be given. By continuous optimization of the multiprofessional patient meeting, regular workshops for team members and internal continuing education for all staff members, and the installation of a research group for supportive and palliative care we aim at enlarging the present activities and to improve the quality of care. Integrated palliative care in Hematology requires commitment, continuing education and a structural change, which initiates a change of views and modern discussions in medicine as well as in society.

References: 1 Lechner (2012) Medicine 91 (4):195.

Introduction: Since several years SAPV is an established part of the palliative care in Rostock. In Rostock a cross-sectoral network consisting of resident oncologists, physicians of palliative care, the University Medical Center Rostock and the hospital (Südstadtklinikum) exists. This network represents a sustainable solution for the palliative care in Rostock. Exemplarily, the data of the year 2014 will be presented. The ambulant palliative care in Rostock is managed by the network of Ambulant Palliative Care (PNR, Palliativnetzwerk Rostock). In 2014, 288 patients were treated. 51.4% of patients were males and 48.6% of the patients were females. The mean and the median age of all patients was 75 and 77yrs, respectively. The most patients 54% (n = 157) were living at home together with their relatives. The other patients were living alone at home or in a care home. The mean Karnofsky index of all patients was 37.9%, the median of the Karnofsky index was 40%. The mean and median of the ECOG was 2.9 and 3, respectively. The mean time of palliative care was 39 days, and the median was 16 days. The minimum time of care was one day, whereas the maximum of care was 430 days. In 70% of the cases the patients were able to die at home or in a care home. The service frequency of the different members of the network was different, representing the voluntary aspect of this secondary occupation. One of the physicians of the team is still available for 24h to provide an excellent palliative care all the time. Conclusions: The Palliativnetz Rostock is an example for an excellent cross-sectoral, multiprofessional care in palliative medicine. An upcoming problem in ambulant palliative care is the increasing number of aged patients. This requires an optimation of the structure of care. Furthermore, the involvement of the University Medical Center Rostock will give the opportunity to perform health service research.

Disclosure: No conflict of interest disclosed. P578

Integration of palliative care in hematologic wards – Best practice model of an university clinic Letsch A.1, Ahn J.1, Erdmann-Schneider P.1, Schleusener A.1, Özöncel O.1, Sandner J.1, Baldus C.1, Janz M.1, Matthas S.1, Pezzutto A.1, Preisler M.1 Charité, Campus Benjamin Franklin, Med. Klinik III, Berlin, Germany

Hematological malignancies are characterized by heterogenous clinic symptoms, prognoses and therapeutic approaches. Often curative treatment intentions persist over a long period and prevent early integration of palliative care (PC) aspects. How do this conditions enable a early integration of palliative care? A selective literature search in parallel to analyses of clinical resources and structures was the basis for the extension of the weekly multiprofessional patient meetings to all wards of the department for Hematology, Oncology and Tumorimmunology. The professions involved are: Nurses, physicians, social workers, psycho-oncologists, physiotherapists, patient managers and as a innovation in addition a palliative care specialist, a pastoral worker, as well as if required nutrition experts, wound and ostomy care, ambulatory hospice service and voluntary visitation service. The implementation of the palliative complex treatment according to OPS 8–892 happened as smooth and progressive transition. In the setting of the weekly multiprofessional patient meeting all patients on the ward are discussed according to their multidimensional requirements, with specific consideration of their psycho-social and their spiritual supply needs. Careful team evaluations consider the inclusion of single patients into the palliative complex treatment. The attendance of all care team members enables a multidimensional perspective on the situation of patients and their relatives and the development of multiprofessional treatment con-

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Disclosure: Anne Letsch: Financing of Scientific Research: BMS, Merck, Novartis,; Expert Testimony: Celgene, Novartis Martina Preisler: No conflict of interest disclosed. P579

Analysis: SAPV (Specialized Ambulant Palliative Care) in Rostock – results of a successful cross-sectoral multiprofessional care Gläser D.1, Leithäuser M.2, Lakner V.2, Kriesen U.3, Lestin M.1, Große-Thie C.3, Junghanss C.3, Nemitz J.4, Leithäuser C.5, Zellmer L.1, Krammer-Steiner B.1 Klinikum Südstadt und Palliativnetz Rostock PNR, Hämatologie/Onkologie/ Palliativmedizin/Hämostaseologie und Komplementärmedizin, Rostock, Germany, 2Schwerpunktpraxis Hämatologie/ Onkologie/ Palliativmedizin und Palliativnetz Rostock PNR, Rostock, Germany, 3Universtätsmedizin Rostock und Palliativnetz Rostock PNR, Klinik für Hämatologie/Onkologie/Palliativmedizin, Rostock, Germany, 4Palliativnetz Rostock GbR PNR, Rostock, Germany, 5 Pflegeversorgungszentrum Rostock PVZ GbR, Rostock, Germany 1

Disclosure: No conflict of interest disclosed.

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Survey of palliative care patients about the “Patientenrechtegesetz”, about their state of knowledge, the implementation of the law and the patients’ wish for more self-determination Rehklau S.1, Torner J.1, Junghanss C.1, Kriesen U.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany 1

Introduction: In 2013, a new and more precise law on patients’ rights was approved by the German government (“Patientenrechtegesetz”, Gesetz zur Verbesserung der Rechte von Patientinnen und Patienten). The law regulates patients’ information about risks, possible and alternatives treatments and treatment goals. Furthermore documentation responsibilities are governed. Patients in palliative care (PC) setting are particular vulnerable and depending on the caring team in a very special way. We proposed that most patients in PC are not aware of their legitimate rights as people being treated in medical fields. Clarification on rights often takes place on palliative care wards. Here, we report on an ongoing survey study addressing this issue. Method: Survey and data collection are done with the help of a self-designed questionnaire. The questionnaire consists of 43 questions, of which 40 are closed and three open questions. Consecutive inpatients and their relatives on PC ward of University of Rostock were interviewed after clarification and written informed consent between 03/2015 and 4/2016 (ongoing study). The questionnaire is divided into 3 parts: The first part asks for the patient’s state of knowledge. Secondly the implementation of the law is asked for and finally about the patients’ wish for more self-determination. The answer options were designed with the help of the “Linkert Scale” or “YesNo-Answers”. Results: Currently the study includes 45 persons. Only 9% of the interviewed were aware of the law. The implementation of the law in reality varies: 73% asked persons felt that they were informed about their diagnosis in an adequate way. Only 67% stated that they received a good explanation of the expected development of their disease by their physician. 71% answered that they were told sufficiently about therapeutic alternatives. Concerning the prognosis related to the palliative diagnosis and therapy only 62% felt that they were informed adequately. Of the patients at the PC facility 11% said that they do feel restricted in their lifestyle because of medical treatment recommended. As a wish they expressed the need for a better involvement in medical decision making and the reduction of bureaucracy in the health care system. Conclusion: Although this study is ongoing, the preliminary results can be taken as strong indicator that a need for more information about the German legislation and the legal obligations of physicians is existent. Disclosure: No conflict of interest disclosed. P581

E-learning; palliative medicine – A Bavarian alternative to improve the general out-patient palliative provision Vehling-Kaiser U.1, Levin C.2, Christian A.3, Fröstl C.4, Beier M.5, Lodders S.6, Kaiser F.1 Netzwerk Landshut, Landshut, Germany, 2Hospiz München, München, Germany, 3SAPV Erlangen, Erlangen, Germany, 4HSW GmbH, München, Germany, 5Hausarztpraxis, Erlangen, Germany, 6KVB München, München, Germany 1

The latest survey of “Faktencheck Gesundheit” in Germany resulted in 76% of the Germans choosing their home as their favorite place to pass away. Unfortunately just every 5th German was granted this wish. Thanks to the Specialized Ambulant Palliative(Pain) Provision (abbr. SAPV) patients with serious malady can be taken care of without having to leave their homes, but only 10% of the patients (who are close to the end of their life) fulfill the requirements for a SAPV-provision. In 2014 only 30% of the deceased received a palliative medical provision. Additionally strong var-

Abstracts

iances in the preferred location to pass away appeared depending on the different regions and the number of ambulant-active palliative doctors in the nearby area. Even though numerous palliative basic courses are being offered in Germany the number of patients with insufficient palliative provision is relatively high. On the other hand participating at conferences for further education requires an absence of multiple days, which is not possible for most people working at a doctor’s office. On one side the cooperation of the SAPV and the Bavarian Health Ministry enabled the provision of patients at home even with complex symptoms, but the AAPV (General Amblulant Palliative(Pain) Provision) is neither funded nor rewarded accordingly. A new basic course for palliative medicine was set up by the “Kassenärztliche Vereinnigung Bayern” (KVB), “Bayerischen Hausärzteverband e.V”, members of the AAPV and in cooperation with the German Society for Palliative Medicine that is especially tailored to the everyday life in doctor’s offices. This course covers 40 hours consisting of three E-learning-modules, presence classes and a hospitation. The E-learning-modules are accessible by using the KVB’s platform Cura Campus. The presence classes take place on three Saturdays. The hospitation in a regional SAPV team is the aim of the further education training. The course takers are obligated to work on the E-learning-modules before each presence class. The newly acquired knowledge is examined by ten multiple-choice-questions. The presence classes are limited to 24 participants for each session and have the opportunity to hand in own case studies. There are two courses taking place parallel, - North and South Bavaria. Merely a few days after the announcement of the event date both courses were completely booked out. Disclosure: Ursula Vehling-Kaiser: Advisory Role: Advisory Board; AbbVie, Roche Pharma AG, Lilly Deutschland GmbH, Amgen GmbH, GILEAD Sciences GmbH; Financing of Scientific Research: Advisory Board; AbbVie, Roche Pharma AG, Lilly Deutschland GmbH, Amgen GmbH, GILEAD Sciences GmbH Florian Kaiser: No conflict of interest disclosed. P582

Inpatient and outpatient palliative care demands in patients with incurable malignancies Maleike H.1, Düring U.2, Roll C.1, Schmidt N.1, Borchert K.1, Kahl C.1 Klinikum Magdeburg gGmbH, Klinik für Hämatologie, Onkologie und Palliativmedizin, Magdeburg, Germany, 2Klinikum Magdeburg gGmbH, Klinik für Anästhesiologie und Intensivmedizin, Magdeburg, Germany 1

During the course of the disease patients (pts) with incurable malignancies suffer a significant symptom burden. Until now, it is unclear how palliative care is best to integrate in the management of the pts and their families, especially regarding the in- or outpatient setting. In a retrospective chart review we analyzed data of 184 pts in an inpatient (n = 123, 219 admissions) and outpatient (n = 61, 124 visits) setting regarding their palliative care needs. The median age was 67.0 years (yrs, 36–90). Most of pts had gastrointestinale tumors (n = 87), followed by gynecological (n = 30) and pulmonal (n = 15) tumors. Hematological malignancies were seen in 16 pts. Median number of sites of metastasis was 2.0 (1–5). At the inpatient setting, median duration of disease before first admission at the palliative care unit (PCU) was 18.8 month (0–167). An average of 1.64 (1–5) admissions was seen, with a mean duration of 9.5 days. Main symptom at admission was pain (n = 73), followed by deterioration of general health (n = 59) and dyspnea (n = 37). For the majority of pts non-invasive therapy was performed (80%), whereas 20% of pts received invasive procedures (endoscopy n = 20, surgical n = 17, pleural drainage n = 15). Antineoplastic therapy was given in 69% of the pts before first admission at the PCU compared to 41.1% after the end of hospital stay at the PCU, meaning adjustment of therapy goals. 78.8% (n = 97) of pts deceased during the observation period, for 87 pts place of death was known: hospice: n = 7, hospital: n = 40, PCU: n = 12, at home: n= 28. Time before death without chemotherapy was 10.9 weeks. Main reasons for consulting the outpatient setting was pain (n = 22), deterioration of general health (n = 18) and discussion of disease course (n = 15). At the majority of visits (65%) antineoplastic therapy was still given despite out-

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patients palliative care needs of the pts. In the outpatient setting invasive (peritoneal puncture, transfusion) and non-invasive (discussion of disease course) procedures were done. Palliative care needs of pts with incurable malignancies were high in an inpatient as well as in an outpatient setting. Whereas physical symptoms (pain, dyspnea) were the leading cause for admission, deterioration of general health and discussion of disease course were the main reasons for consulting the outpatient setting. The different needs in both settings should be taken into account for an adequate care of the pts and their families. Disclosure: No conflict of interest disclosed. P583

Comparison: Proportion of cancer patients in the SAPV (specialized outpatient palliative care) and the hospice with comparison of the length of stay Drolle H.1, Vehling-Kaiser U.1, Bäuml E.2, Flieser-Hartl M.3, Kaiser F.4 Onkologisch- Pallitivmedizinisches Netzwerk Landshut, Landshut, Germany, Hopsiz Vilsbiburg, Vilsbiburg, Germany, 3Achdorfer Krankenhaus, Landshut, Germany, 4Universitätsklinik Göttingen, Göttingen, Germany 1 2

Introduction: Since January 2012 the hospice Vilsbiburg provides terminally ill people in their last stage of life, according to the principles of palliative homecare. In our SAPV the proportion of cancer patients is at 87%. It should be clarified whether the proportion of patients underlying malignant (basic) diseases reflect the ones in the hospices. Because the clinical picture of oncological patients is complex and influenced by the underlying malignant disease, it is necessary take an internistic hemato-oncologist into account. Method: The number of patients was raised for the calendar years 2012, 2013, 2014 and 2015 the number of patients. There was a division based on gender and age. Furthermore, the patients were divided into different groups when they were registered such as; malignant tumor disease, nervous system disorders, chronic diseases of the kidney, heart, alimentary tract, or pulmonary disease and stroke. Results: Since the existence of the SAPV 422 patients were cared for, of which are/were 222 female and 200 male. The gender difference didn’t show any significance (p = 0.362). 56% over 70 years old, 23% 62–70 years, 13% 51–60 years, 8% below 51 years. Following admission diagnoses were available: 86% malignant tumor diseases, 2% Nervous system disorders, 3% of patients with chronic diseases in the kidney, heart, alimentary tract or lung disease, 2% stroke, 6% other illnesses. 394 (93%) died in the hospice, 15 (4%) were discharged into an environment at home and 5 (1%) were moved to the nursing home again. The average length of a stay in the SAPV was 25 days, in the hospice it was at 28.½ days. Conclusion: The proportion of malignant underlying diseases in the hospice is almost equal to that of the SAPV. So basically this shows that next to a medical palliative care an oncological care in hospices is desirable for patients to achieve optimal therapy concepts. Disclosure: No conflict of interest disclosed. P584

Local and regional case management in palliative medicine improves patient management towards the end of life Respondek-Dryba E.1, Weidner-Zellmer L.1, Gläser D.1, Haack L.1, KrammerSteiner B.1 Klinikum Südstadt Rostock, Klinik für Innere Medizin III, Rostock, Germany

Background: In 2010, a coordination centre for palliative care has been established at the City Hospital Rostock. The coordinator’s main tasks include supervision of the medical palliative consultation service (MPCS), its integration into hospital and hospice-based clinical structures as well as into regional networking structures. In addition, new case management

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tools were established including case finding and outreach strategies. The effectiveness of this approach was evaluated. Patients and methods: Between 10/2010 and 10/2015 the MPCS has been refined. In addition to a physician consultant, a palliative care coordinator is present in the MPCS. The requesting procedure for MPCS based on case finding criteria has been introduced and partners from different professions according to specific requirements have been recruited for integrated care. The key to success in case management is the clarification process resulting in recruitment of the right professional service at the right time for a given patient. Critical points of intersection deserve special attention. Outreach methods have been established to announce and offer MPCS locally (City Hospital Rostock) and within the region in networking structures. The present analysis is based on MPCS provided for 462 patients from 10/2010 to 10/2015. Results: A structured MPCS requesting form based on the presence of a) chronic progressive disease (oncology, internal medicine, neurology) with no curative treatment available, b) pain-related syndrome, and c) palliative signs and symptoms with impact on quality of life (nausea/ vomiting, dyspnoe, ascites). According to the case under consideration, the physician consultant and cordinator were supplemented by qualified nursing staff, physiotherapist, minister, ergotherapist, psycho-oncologist and hospice staff. The main outcome indicators of the new approach were improved quality of life mainly by sufficient pain control, improved overall symptom control, enhanced tolerance to and effect of medical therapy, prolonged survival, safer approach to disease by patient and family members, prolonged stay at home and shorter duration of hospital stays, less requirements of emergency care, and less spending for hospital stays. Conclusions: The establishment of MPCS resulted in integrative, patient-focused, effective and symptom-oriented provision of palliative care, adding to the options of hospital and ambulatory palliative care. Disclosure: No conflict of interest disclosed. P585

AML patients in palliative care setting: Influence on life span and transfusion habits, assessment on quality of life and previous treatment Riedel J.1, Kriesen U.1, Große-Thie C.1, Henze L.1, Gläser H.1, Murua Escobar H.1, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany 1

Introduction: Acute myeloid leukemia patients suffer from variable physical symptoms as well as psychological distress. Whereas in younger patients treatment concepts aim at cure, prognosis in older patients is grave and oncological treatments aim at life prolongation. Integration of palliative care concepts into oncology care of AML patients is not well defined. Here we report on a retrospective pilot study analyzing AML patients that were treated in the palliative care ward of the University of Rostock. Method: Consecutive AML patients that were treated on the interdisciplinary palliative care ward between 08/2008 and 12/2015 were analyzed Analysis included patients´ characteristics like age, type of AML, previous therapy, referral date, duration of hospitalization and place of death as well as transfusion patterns. Aspects of quality of life were assessed orally and by questionnaires including numeric rating scale (NRS). Results: A total of 33 AML patients were cared for. Median age was 77 years (range 54 - 89 years). Most patients were male (n = 20, 61%), less female (n = 13, 49%). Median number of PC ward admissions were 2 (range 1 to 4). Six patients (18%) were initially treated with curative intention, 20 patients (61%) received a palliative chemotherapy with hypomethylating agents and seven patients (21%) received best supportive care. Weakness, loss of appetite and fatigue were reported as the most important physical symptoms. Most patients died inpatient. Places of death were at PC ward (84,5%), at hospice (9%) and at home (6,5%).The time of survival depended on previous therapies, amount of transfusions and characteristics of AML. Of interest, most patients received RBC (20/33, 61%) and platelet transfusions (20/33, 61%) during their stay on PC ward. RBC transfusions

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significantly increased Hb-values as expected, there were no changes in physical well being (dyspnea, weakness, fatigue). Plt.-transfusions did reduce visible bleedings. Conclusions: Our study demonstrates the need for optimized early integration of standard palliative care into oncology treatment concepts of AML patients. In particular transfusion policies and the discussion on it with the patients and their relatives should be addressed in the future. Disclosure: No conflict of interest disclosed. P586

Complementary and alternative medicine (CAM) application in the gyn-oncology focus practice Rotmann A.R.1 Praxis Dr. Rotmann, Frauenheilkunde, Rodgau, Germany

Currently, each year approximately one-half of a million persons develop cancer in Germany, with over 200,000 of them succumbing to the disease. The number of mammary carcinomas developed during the same period of time stands at approximately 75,000. As well-founded studies show, the question an affected person poses first is: What can I do for myself? The question for oncologists to ask themselves should be: Which are the patient´s expectations placed on us?! It is precisely in the “gap” opening between the two that for years now complementary alternative medicine (CAM) has been emerging. Numerous studies reveal that the majority of the affected persons turn to complementary medical methods and that they do so deliberately, a fact evidenced by the results of recent studies. Unfortunately though, many clinically active oncologists ignore this trend. As a result, pacients yield to treatments which are in part questionable and not evidence-based. And yet, the economic potential this field holds is an enormous one, with sales of medicinal products, such as generated by over-the-counter herbal drugs for self-medication going into billions, both in Germany and in the US. Two years ago, AGO IMED was founded at the German Cancer Congress. Since then, integrative therapy methods have been clinically explored and used at various university hotspots throughout Germany. In Germany taken alone, over 15 CAM studies are currently in progress. The pertaining paper comes to give you a rewarding insight into the administration of CAM in a practice specialising in gynaecologic oncology. The objective of the participating oncologists is to raise awareness for the needs, apprehensions, and feelings of the patients, so as to be able to approach them in an empathic and thoughtful manner. Disclosure: No conflict of interest disclosed. P587

Mindfulness-based stress reduction for women diagnosed with breast cancer: A systematic review and meta-analysis Will A.1, Monsef I.1, Wöckel A.2, Skoetz N.1 University Hospital of Cologne, Department I of Internal Medicine, Cochrane Haematological Malignancies Group, Cologne, Germany, 2University Hospital of Würzburg, Department of Gynaecology and Obstetrics, Würzburg, Germany 1

Introduction: Breast cancer diagnosis and therapy is a radical and life-threatening experience affecting patients´ quality of life (QoL) drastically with psychological distress and symptoms such as sleep disorder, depression, anxiety and fatigue. This review is to assess the effects of mindfulness-based stress reduction (MBSR) on women diagnosed with breast cancer and it has been funded by the NIH Cochrane Complementary Medicine Bursary. Methods: We searched Cochrane Central Register of Controlled Trials and MEDLINE (1950 to 3/2016) and conference proceedings for randomised controlled trials (RCTs) including trials comparing MBSR in addition to standard care to standard care only in female breast cancer patients. Two review authors independently screened search results, ex-

Abstracts

tracted data and assessed the quality of trials. We contacted six authors for missing data, but only two sent additional data. Results: We identified nine RCTs involving 1153 patients presenting all stages of breast cancer, receiving active treatment or up to six months after therapy. Four RCTs (n = 519) assessed the efficacy of MBSR in terms of quality of life, but due to missing data only two trials (n = 255) were meta-analysed, resulting in a possible benefit for the MBSR group (SMD 0.31, 95% CI 0.06 to 0.56; p = 0.01). Four trials (n = 353) evaluated fatigue, two trials (n = 255) were meta-analysed and showed that MBSR may improve fatigue (SMD -0.43, 95% CI -0.66 to -0.20; p = 0.0003). Effects on depression (three trials) and anxiety (three trials) could not be pooled due to inconsistent reporting of data, but showed in single trials effect favouring MBSR. Quality of sleep was assessed in five trials (n = 597) and was meta-analysed in two trials (n = 163), however, did not show evidence for a difference (SMD -0.00, 95% CI -0.40 to -0.22; p = 0.56). None of the trials provided data on overall survival (OS) and adverse events (AEs). Conclusions: Only few data is available for being meta-analysed, therefore the quality of the evidence for a benefit in fatigue, quality of life, anxiety and depression is of low to very low quality. No studies were found that looked at OS and AEs. Hence, more raw data and longer follow-up periods are needed to evaluate if these effects also translate into long-term effects. Disclosure: No conflict of interest disclosed. P588

Complementary Medicine in Oncology. Are there indications? Kragl B.1, Große-Thie C.1, Henze L.1, Kriesen U.1, Murua Escobar H.1, Schaich M.2, Junghanss C.1, Lampe H.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany, 2Klinik für Hämatologie, Onkologie und Palliativmedizin, Rems-Murr-Klinikum, Winnenden, Germany 1

Introduction: Diseases or specific situations define indications in which the employment of a given treatment is useful. Complementary treatments are used self-reliant or recommended by 60–80% of tumour patients to improve tumour control or to reduce side-effects. Currently, there are no evidence-based indications when to recommended or avoid complementary approaches. Aim of the study was to develop a strategy when to advice patients to specificly use or avoid complementary treatments during their conventional therapy. Methods: Patients were asked orally about usage of complementary treatments and their motivation for the employment. In case of unusual therapeutic courses (short – long time to progression, unusual side-effects) during established treatment regimens we correlated their respective individual complementary treatment pattern to their course of diseases. Further, a literature review for similar cases and evidence was performed. Results: Our own experience as well as literature search did not identify indications for the use of complementary treatments in context of standard medical treatment. Single substances as well as medical systems show diverse results, when used in combination with chemotherapy (Examples 1, 2). Example 1: Male patient, 58 y with metastasised colon cancer with inadequate response to FOLFOX- and FOLFIRI- therapy. During the course of disease up to 36 different complementary treatments were used. After discontinuation of the complementary treatments the patient responded to Capecitabine and other 5-Fluorouracil combinations. Example 2: Male patient, 55 y, gastric cancer. considered the use of glutathione as complementary treatment in combination with FLOT-Regimen. Literature research revealed some evidence, that glutatione may increase effectiveness and reduce neurotoxicity caused by platinum derivates but preclinical data show increasing inactivation and excretion of platinum derivates and therefore preventing DNA damage. Conclusion: Indications for complementary treatments have to be evaluated and defined for specific treatment situations (diseases, treatments,

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state of the patient). Currently there is no indication to advice complementary treatments for a patient who responds to standard therapeutic protocols as expected and tolerates it with acceptable side-effects. Treatment resistance and severe side-effects are a strong indicator to discuss complementary treatments options and eventual risks with the patient. Disclosure: No conflict of interest disclosed. P589

Usage of Chinese Medicine in Western Oncology Lampe H.1, Feng Y.2, Leung A.W.3, Murua Escobar H.1, Große-Thie C.1, Henze L.1, Kriesen U.1, Schaich M.4, Junghanss C.1, Kragl B.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany, 2School of Chinese Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China, 3School of Chinese Medicine, The Chinese University of Hong Kong, Hong Kong, China, 4Klinik für Hämatologie, Onkologie und Palliativmedizin, Rems-Murr-Klinikum, Winnenden, Germany 1

Introduction: Tumorspecific treatments of Western Medicine (WM) has raised the cure rate for cancer to more than 50% during the last century. Individualised treatment of Chinese Medicine (CM) focuses on the interaction of cancer and treatment with the healthy body. Efficacy of CM has been proven by isolated monosubstances (Indirubin, Irinotecan, Topotecan). Efficiency can be estimated by meta-analyses from combination studies of standard treatment and Chinese decoctions. Method: We did use Chinese Medicine for patients with severe side effects or resistance to standard treatments. For severe side-effects we used Chinese decoction in between conventional treatment cycles. In case of chemotherapy resistance WM and CM were used simultaneously. Patients were evaluated for treatment tolerability and tumor response. The results were compared with the existing literature. Results: In single patients with colon cancer we can demonstrate, that the combination of standard therapy with Chinese decoctions allows continuation of treatment, when side-effects become intolerable for the patient (Example I). Example I: Female patient of 64 years presents to us because of intolerable side-effects with FOLFIRI plus bevacizumab for pulmonary metastases of colon cancer (general state, mucositis, dizziness). We first stabilise her with Chinese decoctions and 6 weeks late combined this with standard therapy (Capecitabine). With the combination of Capecitabine and Chinese decoction she could be stabilised for eleven months. Also in single patients with pancreatic cancer we can demonstrate, that Chinese decoctions are capable to reverse treatment resistance (Example II). Example II: Female patient with pancreatic carcinoma who did respond to Gemcitabine and Capecitabine for 3 months each. The combination of Capecitabine and Chinese decoctions she had clinical and tumor marker response for 9 months. Conclusion: The combination of WM and CM is effective in individual patients to reduce side-effect and to reverse treatment resistance. For colorectal cancer studies from China report reduced relapse rates for stage II and III as well as prolonged overall survival for stage IV cancer. For pancreatic cancer preclinical and clinical studies from China show effectiveness of Chinese decoctions alone and in combination with standard treatments. So far it is not understood if the effects are due to additive effects, interaction or influence on the microenvironment. Disclosure: No conflict of interest disclosed.

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P590

Comprehensive assessment of fatigue syndromes in patients after blood stem cell transplantation Rottorf M.1, Horneber M.1, Dressler S.1, Wendelin K.1, Cebulla S.1, Wilhelm M.1, Schäfer-Eckart K.1 Medizinische Klinik 5, Klinikum Nürnberg, Nürnberg, Germany

Introduction: Long-term complications after hematopoietic stem cell transplantation are common and related with a considerable burden of symptoms and distress. Among them, fatigue belongs to the most prevalent and burdensome. Objective: The aim of the study is to investigate physical, psychological and physiological factors associated with fatigue in patients after stem cell transplantation in a prospective longitudinal study. Methods: Patients (Pts) undergo first assessment in a time frame between 12 months and 3 years after allogeneic tx (allo-tx) or between six months and two years after autologous transplantation (auto-tx). The assessment consists of a standardized set of questionnaires including BFI (Brief Fatigue Inventory), EORTC QLQ-C30 and QLQ-FA13 (core questionnaire and fatigue module), HADS (Hospital Anxiety and Depression Scale) and RS-13 (Resilience Scale). Caseness for fatigue syndroms were defined as BFI average severity/impact-score ≥ 4. Thresholds for symptom burden and/or reduced functioning were: HADS: anxiety > 6, depression >6; QLQ-C30: pain ≥ 25, insomnia ≥ 33, cognitive functioning < 84. This assessement will be followed by a symptom-guided Intervention strategy and further follow-up assessments. Results: 47 pts were enrolled since August 2015, 20 after allo-tx and 27 after auto-tx. Diagnoses have been acute or chronic leukemia (15), myeloproliferative disorders (2), multiple myeloma (2) and malignant lymphoma (1) after allo-tx and multiple myeloma (23) and malignant lymphoma (4) after auto-tx. Eighteen out of 47 patients (38%) experienced fatigue with a moderate to severe impact on daily activities. 13 of them (72%) reported high measures of anxiety and 12 (67%) of depression, 12 significant levels of pain (67%), 12 had sleep disturbances (67%) and 13 cognitive impairments (72%). Patients with fatigue syndromes had a significantly diminished quality of life (mean score 42/100, SD±19). Conclusions: The data of this ongoing study demonstrate the high prevalence of fatigue syndromes, their association with other symptoms and distress and their impact on functioning and quality of life. It underlines the importance of a thorough symptom assessment and implementation of symptom-guided management strategies after transplantation. Data from follow-up assessments will be presented. Funding: Deutsche Fatigue-Gesellschaft, Köln Disclosure: No conflict of interest disclosed. P591

SENSe-Study interim analysis structured evaluation of sustainability of sports after cancer Roggenhofer S.1, Widmann T.2 ACURA Waldklinik, Onkologie, Dobel, Germany, 2GKB Klinikbetriebe GmbH Asklepios Klinik Triberg, Onkologie, Triberg, Germany 1

Introduction: In tumor patients who exercise 18–27 metabolic equivalents per week (MET), disease reccurency and potentially mortality after cancer may be reduced. The presented study adresses the question if patients achieve a prognostic relevant level of exercise during a 3 week inhouse medical rehabilitation and if an exercise diary/ training plan helps to maintain the correct “dosage” of activity thereafter? Methods: We included n = 340 patients in a prospective, randomized, controlled, blinded study. The cohort was divided into two groups (A/B). Both are interviewed about their previous physical activity (using a standardized questionnaire ;GermanPAQ50+). During in-house rehabilitation patients received an exercise diary. Furthermore Group A receives an exercise diary/training plan for the post in-house trial perios (6 months).

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Group A and B both are interviewed via questionnaires regarding their amount of exercise 3 and 6 months post rehabilitation. Results: Patients achieved a mean of 22.47 MET/week (SD = 25.71) of physical activity before rehabilitation. During rehabilitation, patients increased their amount of exercise to a mean of 38.52 MET/ week (SD = 15.90, p < 0.001). 3 months post rehabilitation (n = 236) a mean of 41.06 MET/week (SD = 31.56, p < 0.001) and after 6 months (n = 186) a mean of 43.37 MET/week (SD = 33.04, p < 0.001) and thus had a significantly higher score. Post-hoc comparisons revealed that after 6 months patients in group A manages to exercise a greater amount of MET per week (M = 55.69, SD = 37.02) than group B (M = 36.32; SD= 28.37; p < 0.001). Furthermore we compared paper and pencil documented MET to electronically recorded MET by an activity sensor (SenseWear®). Activity sensor recorded METs mimicked patients’ diaries to a great extend (p = 0.012). Conclusions: Patients do exercise before rehabilitation, however below recommended activity levels. During inhouse rehabilitation, patients reach prognostic relevant levels of physical exercise and can be guided to keep this level after rehabilitation. Patients receiving additional support by training plan/exercise diary can maintain higher activity level even 6 months after rehabilitation. Overall, the setting of an inhouse oncology rehabilitation is able educate and motivate patients to establish and maintain a prognostic relevant level of physical acitivy and therfore may help to reduced cancer morbidity and mortality. Disclosure: No conflict of interest disclosed.

Posterdiskussion

assays showed a significantly better response of ovarian cancer cells with a high SKI expression versus low to a treatment with HDACi or HDACi/ ATO. Moreover, SNON upregulation by ATO could be reverted via VPA co-treatment of the cells. Conclusion: We show that a high SKI expression is correlated with a significantly better response of ovarian cancer cells to HDACi. HDACi treatment also reversed the upregulation of SNON upon ATO treatment. Hence, HDACi offer a new option in the treatment of ovarian cancer overexpressing SKI. Moreover, a combined HDACi/ATO treatment may overcome the SNON-derived drug-resistance mechanism. Hence, SKI and SNON may be used as markers for the treatment of human ovarian cancer with HDACi. Disclosure: No conflict of interest disclosed. P593

Differential expression pattern of 3rd generation checkpoint molecules LAG3, TIM3, and VISTA in ovarian cancer with regard to grading and molecular subtypes Ochsenreither S.1,2, Sigler C.2, Arsenic R.3, Sengül B.3, Kühl A.A.4, Braicu E.5, Dietel M.3, Sehouli J.5, Keilholz U.1,2 Charité - Universitätsmedizin Berlin, Comprehensive Cancer Center, Berlin, Germany, 2Charité - Universitätsmedizin Berlin, Campus Benjamin Franklin, Klinik für Hämatologie, Onkologie und Tumorimmunologie, Berlin, Germany, 3Charité - Universitätsmedizin Berlin, Institut für Pathologie, Berlin, Germany, 4Charité Berlin, Campus Benjamin Franklin, Medizinische Klinik für Gastroenterologie, Infektiologie und Rheumatologie, Berlin, Germany, 5Charité Universitätsmedizin Berlin, Klinik für Gynäkologie, Berlin, Germany 1

Introduction: SKI and its paralogue SNON are oncogenes overexpressed in various solid tumors and leukemias (Deheuninck & Luo, C Res 2009; Bonnon & Atanasoski, CTR 2012). They are part of a repressor complex recruiting histone deacetylases (HDAC) (Stroschein, Science 1999; Nomura, Gen Dev 1999). We have previously shown that a block of myeloid differentiation due to a high SKI expression is partially revertable by an HDAC inhibitor (HDACi) and that SKI expression is inhibited by miR29a (Ritter, Leuk. 2006; Teichler, Blood 2011). In keeping, in ovarian cancer inhibition of miR29 is associated with increased cisplatin resistance (Yu, IJC 2013). In addition, SNON is associated with arsenic trioxide (ATO)-induced drug resistance in ovarian cancer (Kodigepalli, FEBS L 2013) Hence, HDACi could provide a treatment option in drug-sensitive and -resistant ovarian cancer with an increased SKI and SNON expression. Methods: In silico survival analyses of treated patients were evaluated via kmplot.com. SKI and SNON expression were compared via Westernblot in ovarian cell lines and primary cultures. Cells ± SKI were treated with HDACi (VPA, LBH589) and growth curves were performed. Viability assays ± VPA/ATO were performed with cells ± SKI. SKI and SNON were downregulated via RNAi and Annexin V apoptosis assays were performed. Results: In silico analyses showed a significantly worse survival of surgically and cisplatin-treated patients with a high SKI expression versus low expression. In vitro experiments showed ovarian cell lines and primary cultures with high or low SKI and SNON levels. In an ovarian cancer cell line with high SKI and SNON expression downregulation of SKI or SNON via RNAi resulted in an increased apoptosis. Growth curves and viability

Introduction: High-grade serous ovarian cancer (OC) is the most common cause of cancer-related death among women world-wide. Checkpoint inhibitors blocking the PD-1/PD-L1 interaction show activity in OC only in a subgroup of patients. New checkpoint molecules TIM3, LAG3 and VISTA are candidates to be targeted together with PD-1/PD-L1 to enhance clinical efficacy. Aim of the study was to characterize the expression of TIM3, LAG3 and VISTA in advanced OC with regard to biological and clinical features, and to compare their expression patterns with the ones of PD-1 and PD-L1. Methods: Immunohistochemistry was performed on samples of 72 patients with advanced OC, who underwent resection and platinum-based chemotherapy. For analysis, only tumor cells were considered. Staining intensity (weak, moderate, strong) and percentage of positive cells of TIM3, LAG3, VISTA, PD-1, PD-L1 were assessed independent of subcellular staining pattern. Microarray data of OC samples from the TCGA network was used as independent data set. Cox regression and non-parametric tests were used for statistics. Results: TIM3-positivity in 100% of tumor cells was associated with prolonged PFS after chemotherapy (p = 0.036, median PFS 14.7 vs 27.4 months). Staining intensities and percentages of positive cells of LAG3, VISTA, PD-L1 and PD-1 had no impact on PFS or OS. LAG3 staining intensity was highly correlated to PD-L1 (p < 0.001). Reanalyzing the TCGA data, we found differential expressions of TIM3, LAG3, VISTA, and PD-L1 among the molecular subtypes as defined by Tothill RW (Clin Cancer Res, 2008). TIM3 associated with subtype C1 (high stromal response), LAG3, VISTA and PD-L1 with C2 (high immune signature). VISTA showed significant higher protein expression (p = 0.017) in grade 3 compared to grade 2 OC. TCGA data revealed higher VISTA mRNA expression in serous compared to endometroid OC (p = 0.036). Conclusion: LAG3 expression is closely correlated to PD-L1, and an additional blockade of LAG3 might act synergistically to PD-1 or PD-L1 blockade. TIM3 is associated with a different molecular subtype than PDL1 and LAG3. The mechanism behind its predictive value on PFS after platinum based chemotherapy requires further investigation. The mode of action of VISTA differs from LAG3 and TIM3. Its expression is strongly associated with grade 3 serous OC. All three qualify for potential combination partners for anti-PD-1/PD-L1 immune therapy to improve its clinical efficacy or broaden its applicability.

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–192

Gynäkologische Tumoren, Mammakarzinom P592

Histondeacetylase inhibitors as an effective treatment for ovarian cancer cells with high expression of the oncoproteins SKI and SNON Frech M.1, Waanders M.1, Stabla K.1, Reinartz S.2, Neubauer A.1 Clinic for Hematology, Oncology, Immunology and Center for Tumor Biology and Immunology, Philipps University, Marburg, Germany, 2Clinic for Gynecology, Gynecological Oncology, Endocrinology and Center for Tumor Biology and Immunology, Philipps University, Marburg, Germany 1

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Disclosure: Sebastian Ochsenreither: Financing of Scientific Research: BMS, Novartis, MSD Ulrich Keilholz: No conflict of interest disclosed. P594

Small cell carcinoma of the ovary, hypercalcaemic type- a rare differential diagnosis of hypercalcaemia Lestin M.1, Reimer T.2, Gläser D.1, Dieterich M.2, Krammer-Steiner B.1 Klinikum Südstadt Rostock, Klinik für Hämatologie, Onkologie und Palliativmedizin, Rostock, Germany, 2Universitätsfrauenklinik und Poliklinik am Klinikum Südstadt Rostock, Rostock, Germany 1

Ovarian small cell carcinoma of the hypercalcaemic type is a very rare disease. This highly aggressive tumour is associated with hypercalcaemia in two-third of patients. The average age of presentation are woman in their early twenties. Long term survivial is not expected, most patients die within 2 years. SMARCA4 mutation appears to be a major driver of SCOCHT tumorigenesis. Case report: An 18-year old young woman presented with nausea and vomiting due to hypercalcaemia. Parathormone was suppressed. NSE and CA 125 values had been in normal range. The CT-scan showed large abdominal mass but no osteolytic bone lesions. Surgery (adnexextirpation, omentectomy, appendectomy) was performed. Histologically the diagnosis of ovarian small cell carcinoma of hypercalcaemic type was confirmed (pT1a (16cm) pNx V0 L0 Pn0 R0 G3 cM0). Immunhistochemical studies on the tumour cells showed a complete loss of SMARCA4/BRG1 expression. Additionally mutation analysis was done. There was no positive family history for this special tumour type. After surgery the patient was treated with VPCBAE (including cisplatin, cyclophosphamide, doxorubicin, etoposide) regimen. Under supportive therapy the regimen was well tolerated. There is only limited information on treatment and prognosis available. Up to date in early stages operation in combination with adjuvant chemotherapy is recommended. The limited data suggests combination of cisplatin and etoposide seem to be effective due to the fact that all long-term survivors were treated with this agents. The role of radiotherapy alone or in combination with chemotherapy is not clear yet. An international data base including all relevant information would be desirable to improve understanding and hopefully treatment in patients with SCOCHT. Disclosure: No conflict of interest disclosed. P595

DVL3 in breast cancer and tumor progression Schubert A.1, Bleckmann A.1,2, Pukrop T.3, Binder C.1 Universitätsmedizin Göttingen, Hämatologie und Medizinische Onkologie, Göttingen, Germany, 2Universitätsmedizin Göttingen, Institut für Medizinische Statistik, Göttingen, Germany, 3Universitätsklinikum Regensburg, Klinik und Poliklinik für Innere Medizin III, Regensburg, Germany 1

WNT signaling plays a crucial role in developmental processes and diseases such as cancer. We recently revealed activation of β-Catenin independent WNT signaling in cerebral metastases of breast cancer patients (Klemm F et al., Carcinogenesis 2011) and demonstrated prognostic relevance of a β-Catenin independent WNT signaling score including Dishevelled 3 (DVL3) in liver metastases (Bleckmann A et al., Clin Exp Metastasis 2016). DVL has been described as a key player in both β-Catenin dependent and independent WNT pathways. In humans three different subtypes have been identified containing three highly conserved domains: DIX (necessary for β-Catenin signaling), PDZ and DEP (conducting PCP and WNT/ Ca2+ signals). It is still largely unclear how DVL discriminates between these sub-pathways and how this influences breast cancer invasion and metastasis. To clarify this question, we overexpressed DVL3 wild type (wt) and various DVL3 knock-out constructs in the luminal A cell line MCF7. In Boyden

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chamber assays we could show DVL3’s pro-invasive potential. This was not reliant on the DIX dependent formation of dynamic aggregates. Interestingly, the overexpression of DVL3 (wt) even inhibited migration in the plane. DVL3 ΔDIX, though reducing the activation of β-Catenin, was still pro-invasive. Overexpression of DVL3 ΔDEP did not increase invasiveness indicating that the DEP domain is involved in this process. Next, we analyzed the amount and localization of DVL3 in immunohistochemical stainings of human cerebral metastases samples (N = 31). Part of these metastases showed DVL3 only in the cytoplasm, another part both in cytoplasm and nucleus of the tumor cells. However, there was no significant correlation between DVL3 localization and the overall survival. Our results underline the significance of DVL3 and its DEP domain in breast cancer progression. In vitro DVL3 is able to enhance the invasion of a weakly invasive breast cancer cell line independent of β-Catenin. The fact that DVL3 localization in brain metastases was not correlated with survival indicates that WNT signaling is governed by additional factors in vivo. Disclosure: No conflict of interest disclosed. P597

Efficacy and gene expression results from SOLTI1007, a phase II study evaluating neoadjuvant eribulin in hormone receptor (HR)-positive/HER2-negative breast cancer Wuerstlein R.1, Prat A.2, Ortega V.2, Pare L.2, Galvan P.2, Nuciforo P.2, Oliverira M.2, Amillano K.2, Lopez R.2, Lluch A.2, Morales S.2, Gonzales R.2, Manso L.2, Martinez J.2, Llombart A.2, de la Pena L.2, di Cosimo S.2, Rubio I.2, Baselga J.2, Cortes J.2, Harbeck N.3 Klinikum der Universität München, Brustzentrum, Muenchen, Germany, Klinik Vall d `Hebron, Barcelona, Spain, 3Klinikum der Universität München, Muenchen, Germany 1 2

Background: Eribulin is a non-taxane, microtubule dynamics inhibitor approved for the treatment of patients with metastatic breast cancer. To date, no biomarker exists to prospectively select patients who will derive the maximum benefit from this chemotherapeutic. In this study, we explored, the efficacy and the association of pre-treatment expression of mRNA with response in patients with HER2-negative breast cancer treated with neoadjuvant eribulin. Methods: SOLTI1007 is a phase II, open-label, single-arm, exploratory pharmacogenomic study of single agent eribulin as neoadjuvant treatment for stage I-II HER2-negative breast cancer (n = 100 HR+ and n = 100 HR-negative). Patients received 1.4 mg/m2 of eribulin mesylate intravenously on Days 1 and 8 every 21-day cycle, for 4 cycles. Baseline, C2D1, and post-treatment (surgical) tissue samples were collected and gene expression profiled. The PAM50 genes and 492 additional breast cancer-related genes were evaluated using the nCounter platform. Results: 92 evaluable patients were included. The overall pCRb rate was 5.4%. Distribution of the intrinsic subtypes was as follows: Luminal A (39.1%), Luminal B (37.0%), Basal-like (10.9%), Normal-like (10.9%) and HER2-enriched (2.2%). pCRb rates differed significantly by subtype (p = 0.009): Luminal B (11.7%, 4/34), HER2-enriched (50%, 1/2), Luminal A (0%, 0/36), Basal-like (0%, 0/10), Normal-like (0%, 0/10). pCRb rate differed significantly by ROR-P (p = 0.018): ROR-P high (16.7%, 4/24), ROR-P med (1.9%, 1/52), ROR-P low (0%, 0/16). Ki67% did not predict pCRb (p = 0.406). Subtype change at surgery occurred in 48.3% (14/29) of Luminal Bs, 13.8% (4/29) of Luminal As, 100% (2/2) of HER2-enriched and 12.5% (1/8) of Basal-like tumors. 78.6% (11/14) of subtype changes in Luminal B disease were to Luminal A. Compared to screening biopsies, 136- and 141- genes were found up- and down-regulated in surgical specimens. The up-regulated gene list was enriched for response to hormone stimulus, negative regulation of apoptosis and angiogenesis. The down-regulated gene list was enriched for cell cycle, DNA-repair and microtubule cytoskeleton organization. Conclusions: From a response and biological perspective (i.e. induction of a Luminal A phenotype), patients with Luminal B disease might benefit

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the most from eribulin therapy. Strategies combining eribulin with endocrine therapy seem warranted in Luminal B breast cancer. Disclosure: Rachel Wuerstlein: Financing of Scientific Research: für Vorträge und Beratertätigkeit Eisai Nadia Harbeck: Expert Testimony: Studien in Zusammenarbeit mit Eisai

P599

Endurance and resistance training with breast cancer patients during chemotherapy – a clinical intervention trial Schmidt T.1, Keller L.2, Dürkop J.2, Jonat W.2, Weisser B.3, Röcken C.1, Mundhenke C.2 Krebszentrum Nord, Universitätsklinikum Schleswig-Holstein, Kiel, Germany, Klinik für Gynäkologie und Geburtshilfe, Universitätsklinikum SchleswigHolstein, Kiel, Germany, 3Institut für Sportwissenschaft, Christian-AlbrechtsUniversität, Kiel, Germany 1 2

P598

Efficacy and safety of nab-paclitaxel in patients with metastatic breast cancer: Final results of the noninterventional study NABUCCO Marschner N. , Potthoff K. , Salat C. , Söling U. , Hansen R. , Grebhardt S. , Harde J.2, Nusch A.6 1

Praxis für interdisziplinäre Onkologie & Hämatologie, Freiburg i.Br., Germany, iOMEDICO AG, Freiburg i.Br., Germany, 3Gemeinschaftspraxis für Innere Medizin, Hämatologie u. Internistische Onkologie, München, Germany, 4 Onkologische Gemeinschaftspraxis Dres. Söling/Siehl, Kassel, Germany, 5 Onkologische Schwerpunktpraxis Dres. Hansen & Reeb, Kaiserslautern, Germany, 6Praxis für Hämatologie und internistische Onkologie, Ratingen, Germany 1 2

Introduction: One of the most effective chemotherapies for metastatic breast cancer (MBC) is nab-paclitaxel (nab-P) which is approved for the treatment of MBC after failure of 1st-line therapy and when anthracyclines are not indicated. Randomized clinical trials (RCT) have shown high efficacy and acceptable toxicity. Real world data of nab-P in MBC, however, are still limited. Methods: The prospective multicenter non-interventional study NABUCCO was designed to collect data on the routine treatment of 700 patients (pts) with MBC in approximately 100 sites across Germany. Primary objective was the time to tumor progression (TTP), secondary objectives were overall response rate (ORR), overall survival (OS), the dosage scheme of nab-P, time on treatment, safety parameters and quality of life. Descriptive statistics were used to analyze the data. TTP and OS were calculated using the Kaplan-Meier method. Results: Between 4/2012 and 4/2015 705 pts with MBC at 128 active sites had been enrolled. 697 patients were evaluable with a median follow-up of 17.7 months. Baseline characteristics: Median age 62.3 years (range 29.2– 89.3), age ≥ 65 years n = 291 (41.8%), ECOG PS ≥ 2 n = 49 (7.0%), prior taxanes 419 pts (60.1%). Pts were treated at the physician’s discretion. The application mode of nab-P was as follows: 260 mg/m² q3w (n = 153, 22.0%), ≥ 15% reduced dose q3w (n = 37, 5.3%), 150 mg/m² d1, d8, d15 q4w (n = 54, 7.7%), ≥ 15% reduced dose d1, d8, d15 q4w (n = 219, 31.4%), 100–125 mg/m2 d1,8,15 q3w (n = 90, 12.9%) and other (n = 144, 20.7%). Pts received nab-P as 1st-line (n = 280, 40.2%), 2nd-line (n = 169, 24.2%), 3rd-line (n = 141, 20.2%) or ≥ 4th line (n = 107, 15.4%) therapy. Median TTP for all pts was 5.9 months (95% CI 5.6–6.4), with a median TTP of 7.1 months (95% CI 6.0–8.6) for 1st-line, 6.0 months (95% CI 5.5–7.3) for 2nd line, 5.6 months (95% CI 4.6 -6.7) for 3rd line and 5.2 months (95% CI 4.2–5.6) for ≥ 4th line treatment. ORR was 37.2% for all pts, 46.1% for 1st line, 30.2% for 2nd line, 31.9% for 3rd line, and 29.0% for ≥ 4th line treatment, respectively. 34.3% of patients developed adverse events grade 3/4 which included leukopenia (7.9%), peripheral sensory neuropathy (4.3%) and infections (4.2%). Further subgroup analyses will be presented. Conclusions: The results of the NABUCCO study confirm the clinical trial outcomes and the favorable benefit-risk profile of nab-P in patients with MBC in a real-life setting. Disclosure: No conflict of interest disclosed.

Abstracts

Previous findings suggest that physical activity during breast cancer treatment can reduce side-effects and improve clinical outcome. In the present study, endurance (ET) and resistance training (RT) in 67 patients with breast cancer undergoing chemotherapy were compared with standard care (SC). Method: Study endpoints were muscular strength (NM) at latissimus pull down, bench press and leg press, endurance (Watt/kg/bodyweight in kg) and subjective perceived exertion during the endurance stress test (Borg scale) and quality of life (QoL) measured by standardized report form of the European Organization for Research and Treatment of Cancer (EORTC QLQ C30+BR23) before and after 12 weeks of treatments. Results: Patients in the RT and ET improved significantly in bench press (RT: p = 0.021; ET: 0.023), latissimus pull down (RT: p = 0.014; ET: n.s.) and there was a trend in leg press (RT: n.s.; ET: n.s.) strength compared to the SC. All groups decreased in the endurance-stress-test (RT: p: n.s; ET: p: n.s; SC: p: n.s.), however the maximum endurance lost in Watt appeared in the SC group (p: 0.001). The subjective perceived exertion at 100 watts remained stable in the RT (p: n.s.), decreased the most in SC (p: n.s.) and to a lesser extend in the ET group (p: 0.02 ). In the RT group QL improved significantly (p: 0.011). There is also a trend for improvement of QL in the ET group (p: >0.05; n.s.) whereas the SC group showed a decrease in QL. Conclusion: The results highlight improvements in strength, endurance and QL from exercise training and support its implementation into standard of care during chemotherapy for breast cancer patients. All authors declare no conflict of interest. Disclosure: No conflict of interest disclosed. P600

Integration of genomics and histology reveals diagnosis and effective therapy of refractory cancer of unknown primary with PDL1 amplification Gröschel S.1, Bommer M.2, Hutter B.3, Budczies J.4, Bonekamp D.5, Heining C.1, Fröhlich M.3, Hübschmann D.3, Geörg C.3, Richter D.3, Pfarr N.6, Pfütze K.3, Wolf S.3, Schirmacher P.7, Jäger D.1, von Kalle C.1, Weichert W.1, Stenzinger A.7, Fröhling S.1, DKTK NCT Heidelberg, Heidelberg, Germany, 2Klinikum am Eichert, Department of Hematology, Oncology and Infectious Diseases, Göppingen, Germany, 3DKFZ Heidelberg, Heidelberg, Germany, 4Institute of Pathology, Charité University Hospital, Berlin, Germany, 5Department of Radiology, DKFZ, Heidelberg, Germany, 6Institute of Pathology, Klinikum rechts der Isar, Technische Universität München, München, Germany, 7Institute of Pathology, Heidelberg University Hospital and NCT Heidelberg, Heidelberg, Germany 1

Identification of the tissue of origin in cancer of unknown primary (CUP) poses a diagnostic challenge and is critical for directing site-specific therapy. Currently, clinical decision making in patients with CUP primarily relies on histopathology and clinical features. Comprehensive molecular profiling has the potential to contribute to diagnostic categorization and, most importantly, guide CUP therapy through identification of actionable lesions. We here report the case of an advanced-stage malignancy initially mimicking poorly differentiated soft-tissue sarcoma that did not respond to multi-agent chemotherapy. Molecular profiling within a clinical whole-exome and transcriptome sequencing program revealed a heterozygous, highly amplified KRAS G12S mutation, compound-heterozygous TP53 mutation/deletion, high mutational load, and focal amplification of chromosomes 9p (including PDL1 [CD247] and JAK2) and

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10p (including GATA3). Integrated analysis of molecular data and conventional histopathology suggested a diagnosis of triple-negative breast cancer (TNBC) and provided a rationale for immune checkpoint inhibitor therapy with pembrolizumab, which resulted in rapid clinical improvement and a lasting partial remission. Analysis of 157 TNBC samples from The Cancer Genome Atlas revealed focal, high-level PDL1 amplification coinciding with excessive PDL1 mRNA expression in 24% of cases. Collectively, these results illustrate the diagnostic utility of multi-dimensional tumor profiling in cases with non-descript histology and immune phenotype, demonstrate the predictive power of genomic PDL1 amplification for immune checkpoint inhibition, and suggest a targeted therapeutic strategy in chromosome 9p24.1/PDL1-amplified cancers. Disclosure: No conflict of interest disclosed. P601

Effects of a phone-based follow-up care after indoorrehabilitation for breast cancer patients – A randomized controlled trial Hass H.G.1, Stepien J.1, Lerch J.1, Schröck R.1, Berger D.2, Tripp J.3 Paracelsus-Klinik, Internistische Onkologie, Scheidegg, Germany, Arbeitsgemeinschaft für Krebsbekämpfung NRW, Bochum, Germany, Universität Münster, Institut für Medizinische Psychologie, Münster, Germany

1 2 3

Background: To evaluate feasibility and long term effects of a telephone-based follow-up intervention after oncological rehabilitation. Methods: 172 breast cancer patients (age 27–54 years) were randomized after inpatient rehabilitation to a telephone-based intervention (phone calls every 4 weeks over 6 months) or control group. Patients were evaluated by standardized questionnaire at T1 (start of rehabilitation), T2 (end of rehabilitation) and T3 (six months after rehabilitation) including IRES24 and HADS questionnaire. Results: 2-way-ANOVAs were performed to evaluate long-term effects. Main effects of IRES-24 and HADS were significant depending on time (IRES-24 F(2,116)=40,49, p < .01 and HADS F(2,117)=31,50, p < .01; (F(2,11 6)=31,19, p < .01) but no significant differences between the intervention and control group were seen. Conclusions: Telephone-based follow-up care is feasible with high patient acceptance. However an improvement of long-term effects in the intervention group were not be detected by IRES-24 and HADS questionnaire. Potential explanations may be the low „dosage” (duration/quantity of phone calls) of the intervention or the fact that in the last years multimodal treatment interventions were established in german rehabilitation centers leading to a so-called “ceiling effect” without significant effects of additional follow-up interventions. Disclosure: No conflict of interest disclosed.

Disclosure: Hans Tesch: Employment or Leadership Position: Niedergelassener Onkologe im Centrum für Hämatologie und Onkologie Bethanien Frankfurt.; Advisory Role: Novartis, Roche; Financing of Scientific Research: Novartis, Roche, Pfizer, Eisai C. Kurbacher: Advisory Role: Amgen, NewCo, Riemser, Teva; Financing of Scientific Research: Amgen, Astra Zeneca, Axios, Hexal, Novartis, Roche, Teva, Zytoservice P603

Oncologists’ letters for breast cancer patients Baumann W.1, Schüssler L.2, Bertram M.3, Benser J.1, Kümpers S.2, Hermes-Moll K.1 WINHO GmbH, Köln, Germany, 2Hochschule Fulda, Fachbereich Pflege und Gesundheit, Fulda, Germany, 3Hämatologisch Onkologischer Schwerpunkt, Hamburg, Germany 1

P602

Health service reality of therapy management in hormone receptor positive advanced breast cancer patients: collection of retrospective data at 17 German sites Tesch H.1, Abenhardt W.2, Depenbusch R.3, Fischer D.4, Göhler T.5, Hempel D.6, Hielscher C.7, Hutzschenreuter U.8, Kaiser S.9, Körbel O.9, Kuhn T.10, Schichtl T.11, Ulshöfer T.12, Kurbacher C.13 Centrum für Hämatologie und Onkologie Bethanien, Frankfurt, Germany, Onkologie Elisenhof, München, Germany, 3Onkologische Schwerpunktpraxis, Gütersloh, Germany, 4Klinikum Ernst von Bergmann, Postdam, Germany, 5 Onkozentrum, Dresden, Germany, 6Onkologisches Zentrum, Donauwörth, Germany, 7G.SUND Gynäkologie Kompetenzzentrum, Stralsund, Germany, 8 Onkologische Gemeinschaftspraxis, Nordhorn, Germany, 9Novartis Pharma GmbH, Nürnberg, Germany, 10Gynäkologische Onkologie, Stuttgart, Germany, 11Abteilung Onkologie am MVZ Weiden GmbH, Weiden, Germany, 12 Schwerpunktpraxis für Hämatologie und Onkologie, Ludwigsburg, Germany, 13 Gynäkologische Onkologie, Medizinisches Zentrum, Bonn, Germany 1 2

Introduction: Therapy (TX) management of the new oral therapies for advanced breast cancer (aBC) is challenging for patients (pts) as well as

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physicians. TX management includes not only mitigation of side effects but optimal and holistic time consuming consultation and care of pts. Counseling options of interdisciplinary teams are available for pts to cope with daily life and oncological trained personnel like onco- or breast-carenurses are getting more important in supporting TX management. This study evaluates implementation and usage of additional counseling options in daily practice. Methods: Retrospective data of 639 pts with hormone receptor positive aBC treated in 2014 was collected at 17 German oncological centers including demographics, TX management, and compliance. The questionnaire on side effect management focused on incidence of stomatitis, rash, fatigue, emesis, and pneumonitis in pts treated with chemotherapy (CT), endocrine therapy (ET), and targeted therapy (TT), and education on prophylactic measures. To ensure data privacy aggregated data was collected from sites only. Results: 136 pts treated with systemic therapies suffered from stomatitis (CT: 55,9%, ET: 19,9%, TT: 27,9%), 138 from rash (CT: 45,7%, ET: 13,8%, TT: 11,6%), 225 from fatigue (CT: 56,2%, ET: 22,6%, TT: 8,1%), 179 from emesis (CT: 70,4%, ET: 14,5%, TT: 8,4%), and 46 from pneumonitis (CT: 19,6%, ET: 10,9%, TT: 28,3%). Education on prophylactic measures was performed in nearly 100% of pts. 94,1% of treating physicians planned a closer follow-up at therapy start. To support TX management, onco-nurses were employed at 15 sites and breast-care-nurses at 5 sites. Additional counseling options were offered to nearly all pts (psycho-oncology: 82,4%, physical exercise: 100%, self-help group: 94,1%, nutrition: 88,2%). For about one third of pts no information was available whether one of these measures was used by pts. Monitoring of compliance was mainly done by interview. New tools, like reminders via phone or apps are rarely being offered. Side effects and fear of side effects were reported to be the main reason for noncompliance. Conclusion: The results show the complexity of TX management in relation to therapy and side effects. Education on TX management and prophylactic measures is standard in routine practice; however, systematic monitoring of implementation is only done sporadically. The usage of new tools like apps, reminder SMS, or emails warrants further research.

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Introduction: Breast cancer patients have a distinct demand for information. Unmet needs depend on the patients’ age and health literacy. Our aim is to analyze acceptance, influence, benefit, and suitability of forwarding copy letters to breast cancer patients. Methods: Three office-based oncologists were invited to ask three of their breast cancer patients to participate who did not yet obtain a copy letter. Breast cancer patients with different educational background, disease severity, line of therapy, and age received a copy from their oncologists. All participants were queried either by means of problem-centered or by expert interviews, respectively. Qualitative content analysis after Philip Mayring was used for the evaluation. Results: The supplemental offer was accepted and rated positively by all nine breast cancer patients. Their individual benefit was e.g. the possibility to read independently and to compare at all times, and to show the copy letter to their families. Though not understanding some parts of it, the majority rated the content as predominantly comprehensible. The posi-

Abstracts

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tive attitude towards their office-based oncologists was stable throughout receiving the copy letter. Offer and request for copy letters were handled differently by the three participating oncologists. From the majority’s perspective, this tool may be insufficient to inform adequately their patients, e.g. because of medical terminology. In the opinion of some of the oncologists, benefit and suitability depend on the individual need for information and the way of dealing with it. Sharing copy letters with their patients improves transparency of the medical activities and can be beneficial for the health care process. Conclusion: While some oncologists assess the copy letters as inappropriate for supplemental patient-oncologist-communication, breast cancer patients regard this tool as predominantly gainful. Oncologists appear to stick to their traditional perspective which perceives the copy letter mainly as a communication tool from doctor to doctor. Due to their individual experience, patients seem to develop an emotional relationship to the copy letter containing information about their disease. Especially for patients dealing actively with their treatment process, copy letters could be a reasonable instrument. Disclosure: No conflict of interest disclosed. P604

Serum carbonic anhydrase IX as predictive marker for efficacy of bevacizumab: a biomarker analysis from the GeparQuinto phase III neoadjuvant breast cancer trial Janning M.1,2, Müller V.3, Vettorazzi E.4, Cubas-Cordova M.1,2, Gensch V.1,2, Ben Batalla I.1,2, zu Eulenburg C.4, Schem C.5, Fasching P.A.6, Schnappauf B.7, Karn T.7, Fehm T.8, Just M.9, Kühn T.10, Holms F.11, Overkamp F.12, Krabisch P.13, Rack B.14, Denkert C.15, Untch M.16, Tesch H.17, Rezai M.18, Kittel K.19, Pantel K.2, Bokemeyer C.1, Loibl S.20, von Minckwitz G.20, Loges S.1,2 Universitätsklinikum Hamburg-Eppendorf, II.Med Klinik, Abteilung für Onkologie, Hämatologie und Knochenmarktransplantation, mit Sektion Pneumologie, Hamburg, Germany, 2Universitätsklinikum Hamburg-Eppendorf, Institut für Tumorbiologie, Hamburg, Germany, 3Universitätsklinikum Hamburg-Eppendorf, Klinik und Poliklinik für Gynäkologie, Hamburg, Germany, 4 Universitätsklinikum Hamburg-Eppendorf, Medizinische Biometrie und Epidemiologie, Hamburg, Germany, 5Universitätsklinikum Schleswig-Holstein, Klinik für Gynäkologie und Geburtshilfe, Kiel, Germany, 6Universitätsklinikum Erlangen, Frauenklinik, Erlangen, Germany, 7Universitätsklinikum Frankfurt, Klinik für Frauenheilkunde und Geburtsmedizin, Frankfurt, Germany, 8Universitätsklinikum Düsseldorf, Klinik für Frauenheilkunde und Geburtsmedizin, Düsseldorf, Germany, 9Onkologische Schwerpunktpraxis Bielefeld, Bielefeld, Germany, 10Klinikum Esslingen, Klinik für Frauenheilkunde und Geburtshilfe, Esslingen, Germany, 11St. Barbara Klinik Hamm-Heessen, Abteilung für Gynäkologie und Geburtshilfe, Hamm, Germany, 12Praxis und Tagesklinik für internistische Onkologie und Hämatologie, Recklinghausen, Germany, 13Klinikum Chemnitz, Klinik für Frauenheilkunde und Geburtshilfe, Chemnitz, Germany, 14Klinikum der Ludwig-Maximilians Universität, Klinik und Poliklinik für Frauenheilkunde und Geburtshilfe, München, Germany, 15 Charité – Universitätsmedizin Berlin, Campus Mitte, Institut für Pathologie, Berlin, Germany, 16HELIOS Klinikum Berlin-Buch, Klinik für Gynäkologie und Geburtshilfe, Berlin, Germany, 17Onkologie Bethanien, Frankfurt, Germany, 18 Europäisches Brustzentrum am Luisenkrankenhaus, Düsseldorf, Germany, 19 Praxisklinik – Krebsheilkunde für Frauen, Berlin, Germany, 20German Breast Group, Neu-Isenburg, Germany 1

Introduction: There is an unmet clinical need for identification of biomarkers to guide selection of patients for treatment with bevacizumab. The purpose of this exploratory study was to investigate predictive potential of carbonic anhydrase IX serum levels (sCAIX) for efficacy of bevacizumab in the neoadjuvant GeparQuinto trial. Methods: sCAIX levels were determined by enzyme-linked immunosorbent assay at baseline and after 4 cycles of neoadjuvant chemotherapy with and without bevacizumab. Correlations between sCAIX level and pathological complete response (pCR;), disease-free and overall survival (DFS, OS;) were assessed with appropriate statistical methods using bootstrapping for robust estimates and internal validation. Results: N = 1189 samples of patients with HER2-negative tumors of whom 577 received bevacizumab were analyzed. sCAIX levels significant-

Abstracts

ly increased after 4 cycles neoadjuvant chemotherapy (NCT) plus bevacizumab indicating modulation by bevacizumab. When treated with NCTalone, patients with low sCAIX levels at baseline were less likely to achieve a pCR compared to those with higher sCAIX levels (12.1% vs. 20.1%, p = 0.012). In patients with low sCAIX levels the addition of bevacizumab significantly improved the chance to achieve a pCR (OR, 2.16; 95%CI, 1.24 to 3.88, p = 0.006; pCR-rates: 12.1% for NCT vs. 20.4% for NCT-B), while patients with high sCAIX levels did not benefit from bevacizumab (OR, 0.89; 95%CI, 0.58 to 1.36, p = 0.590; pCR-rates: 20.1% for NCT vs. 17.0% for NCT-B). DFS was significantly better in patients with higher sCAIX levels when they were treated with NCT alone compared to NCT-B treated patients (HR, 1.47; 95%CI, 1.04 to 2.08; p = 0.028; 5yearDFS: 81% for NCT vs. 73.6% for NCT-B). However, no significant differences in DFS were observed in patients with low sCAIX levels (HR, 0.80; 95%CI, 0.52 to 1.22; p = 0.304; 5yearDFS: 71.4% for NCT vs. 78.5% for NCT-B). There was no correlation for sCAIX levels on treatment results with regards to OS. Conclusion: Warranting further external validation, sCAIX may represent a novel biomarker candidate predicting response to bevacizumab and NCT in primary breast cancer. Disclosure: Melanie Janning: Advisory Role: Sanofi, Teva; Financing of Scientific Research: Takeda; Expert Testimony: BerGenBio, Roche; Other Financial Relationships: Travel Expenses: Amgen, Takeda Sonja Loges: Advisory Role: Lilly, BerGenBio, Sanofi; Financing of Scientific Research: Lilly, BerGenBio,; Expert Testimony: BerGenBio, Roche; Other Financial Relationships: BerGenBio, Lilly

Posterdiskussion

Kasuistiken – Hämatologie P605

Filamentous appearance of Enterobacter cloacae in pleural effusion Metzgeroth G.1, Becker K.-P.2, Gencer D.1, Hofmann W.-K.1, Hastka J.1 III. Medizinische Klinik, Hämatologie und internistische Onkologie, Universitätsmedizin Mannheim, Mannheim, Germany, 2Institut für Medizinische Mikrobiologie und Hygiene, Universitätsmedizin, Mannheim, Germany 1

Introduction: Gram-negative bacteria usually divide by forming a central septum across the cell. In this process, penicillin binding protein 3 (PBP3) plays a crucial role. Beta lactam antibiotics inhibit PBP-3 and prevent the formation of the dividing septum. When exposed to sub lethal antibiotic concentrations, the bacteria can elongate but do not divide and appear as long, slender filaments, resembling fungal hyphae. Methods: 59-year-old male suffered from a metastatic cholangiocarcinoma, diagnosed 14 months before. Stable disease was temporarily reached after four cycles of gemcitabine and cisplatin and lasted for about 6 months. Thereafter, progressive bilateral pleura effusions caused shortness of breath and repeated thoracenteses were necessary. The clinical condition of the patient could, however, not be stabilized and has further deteriorated by pneumonic infiltrates and fever. Antibiotic therapy with Tazobac® (fix combination of the b-lactam antibiotic Piperacillin and the beta-lactamase inhibitor Tazobactam) was started and another thoracentesis was performed 3 days later to manage patient´s dyspnea. Results: Cytological examination of a Pappenheim stained smear of the pleural fluid revealed a mixed cell population, predominantly consisting of neutrophils and a few scattered macrophages and mesothelial cells which correspond to a parapneumonic effusion. The smear was, however, dominated by numerous slender, unbranched and up to 100 µm long blue filamentous structures. The filaments mostly lay free between the neutrophils, some in isolation and some in dense clusters. Filaments were also found intracellularly, coiled within the neutrophils. At first glance, these findings closely resemble fungal hyphae and a fungal infection was initially assumed. Microbiological examination of the effusion, however, excluded a fungal infection and revealed a huge number of gram-negative

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bacteria, identified as Enterobacter cloacae group. Enterobacter cloacae is a rod-shaped, gram-negative, facultative pathogen bacterium, normally 0.3–0.6 µm wide and 0.8 - 2.0 µm long. Conclusion: Gram-negative bacteria treated by b-lactam antibiotics sometimes show an abnormal filamentous morphology, which may resemble fungal hyphae. It is important to be aware of this phenomenon in order to avoid misinterpretation and mistreatment. Filamentation suggests an insufficient antibiotic concentration at the site of infection. Disclosure: No conflict of interest disclosed. P606

Is this myeloma? A patient with elevated IgG serum levels and recurrent hypercalcemia Gundel D.1, Köchel C.1, Einsele H.1, Knop S.1 Universitätsklinikum Würzburg, Medizinische Klinik und Poliklinik II, Würzburg, Germany 1

Introduction: Hypercalcemia is a potentially life-threatening condition with variable clinical manifestation, etiology and treatment. Primary hyperparathyroidism and malignancy account for greater than 90% of cases of hypercalcemia. granulomatous diseases and intoxication with active vitamin D metabolites contribute to a diverse array of less frequent causes of calcium homeostasis derangements. Hypercalcemia in malignancy is often associated with elevated PTHrP-levels but is also commonly seen in patients with multiple myeloma due to secretion of osteoclast activating factors by the tumor cells. Case report: We present a case of a 63-year-old woman with recurrent oligosymptomatic hypercalcemia. The medical history was remarkable with the ingestion of high dose vitamin D for three weeks prior to admission. Physical examination showed splenomegaly. Extensive laboratory evaluation revealed no benign endocrine cause of hypercalcemia (suppressed PTH, normal TSH, low 25-OH-Vitamin D3 as a marker for vitamin D intoxication). There was no increase of PTHrP. Because of hypergammaglobulinemia due to elevated IgG serum levels, multiple myeloma was suspected. Yet, there were no osteolytic bone lesions seen in bone CT scan, bone marrow biopsy showed no monoclonal plasma cells and remarkably hypergammaglobulinemia proved to be oligoclonal. However bone marrow biopsy demonstrated CD20+ lymphocytosis consistent with small lymphocytic lymphoma that was found to have deletion of TP53. CT scans and sonography showed mediastinal and mesenterial lymphadenopathy and splenomegaly. Paraneoplastical elevation of 1,25-dihydroxy-vitamin D3 emerged as the culprit for causing hypercalcemia in this patient. After immediate management of hypercalcemia with hydration, loop diuretics and bisphosphonates, immunochemotherapy with Rituximab and bendamustine was initiated. Subsequently the patient was started on ibrutinib, a Bruton tyrosine kinase Inhibitor. With this therapy, the patient hasn´t experienced any further episodes of hypercalcemia and splenomegaly has resolved. Conclusion: This case illustrates the diagnostic challenges and pitfalls in patients presenting with hypercalcemia. Treating the underlying cause of hypercalcemia is crucial for preventing recurrent and detrimental electrolyte derangements and novel agents can extend the current armamentarium. Disclosure: No conflict of interest disclosed.

P607

Bing-Neel-Syndrome: A case report successful treatment using systemic combination chemotherapy Timmer K.1, Große-Thie C.1, Henze L.1, Freitag S.1, Stubbe F.1, Kriesen U.1, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany 1

Introduction: Bing-Neel syndrome (BNS) is an extremely rare neurologic complication of Waldenström macroglobulinemia (WM). It is associated with the central nervous system (CNS) infiltration by neoplastic lymphoplasmacytoid and plasma cells with or without cerebrospinal fluid (CSF) hyperglobulinemia. We report a case of a 47-year-old man with no history of WM but epilepsy and an intracerebral lesion of unclear genesis. He was diagnosed with Bing-Neel-syndrome based on histopathology of cerebrum biopsy. Case report: In 12/2013, a 45-year-old man with a 3-year history of epilepsy presented to the neurology department with complaints of dizziness, blurred vision, lacking in concentration, inside restlessness and distinct fear and dyspnea. Magnetic resonance imaging (MRI) of the brain showed a lesion intracerebral, right temporal with accumulation of contrast medium of unclear genesis and meningeal enhancement. In 12/2014 a follow-up MRI showed progression of the lesion. In 01/2015 a biopsy was taken by surgical intervention. The histopathology showed an infiltration of the brain by low-grade malignant proliferation of lymphoplasmacytoid cells. Neither biopsy of bone marrow, nor laboratory values showed other manifestation of B-cell-Lymphoma. A lumbar puncture was performed and analysis of cerebrospinal fluid (CSF) revealed an elevated white blood cell count of 10 Mpt/l (normal < 5 Mpt/l) with 100% lymphocytes, total CSF protein of 528 mg/l (normal range 150–450 mg/l), IgM level of 9.62 mg/l (normal < 1 mg/l) and IgG level of 69 mg/l (normal range 2.6–24 mg/l). A diagnosis of Bing-Neel-syndrome was made. Systemic chemotherapy with rituximab-high dose-methotrexate and AraC (analog IELSG 32-Studies; rituximab 375 mg/m² day 0, methotrexate 3500 mg/m² day 1, cytarabine 2000 mg/m² day 3 and 4; 6 cycles in total) was begun. All symptoms resolved quickly within weeks of initiation of systemic chemotherapy. A follow-up MRI scan after 3 cycles of therapy showed a regressive meningeal enhancement. All treatments were well tolerated and patient remained clinical stable since completing his treatments in 07/2015. Conclusions: Our case report demonstrates Bing-Neel syndrome without history of WM and therefore it presents with a different approach for the treatment of BNS: applying successfully 6 cycles of systemic chemotherapy analog the treatment of primary central nervous system lymphoma. The patient sustained complete remission 9 months after discontinuation of therapy. Disclosure: No conflict of interest disclosed. P608

Successful treatment of a patient with disseminated Mycobacterium abscessus infection and MonoMAC syndrome by allogeneic hematopoietic stem cell transplantation Simonis A.1, Müller A.M.1, Grassinger J.1, Müller N.J.2, Pabst T.3, Pachlopnik Schmid J.4, Güngör T.4, Manz M.G.1, Schanz U.1 Universitätsspital Zürich, Klinik für Hämatologie, Zürich, Switzerland, Universitätsspital Zürich, Klinik für Infektionskrankheiten und Spitalhygiene, Zürich, Switzerland, 3Universitätsspital Bern, Universitätsklinik für Medizinische Onkologie, Bern, Switzerland, 4Kinderspital Zürich, Abteilung für Stammzelltransplantation, Zürich, Switzerland 1 2

Introduction: The MonoMAC (monocytopenia and mycobacterium avium complex infection) syndrome is an extremely rare and life threatening genetic disorder caused by loss of function mutations in the GATA binding protein 2 gene, a zinc-finger transcription factor which is crucial in the regulation of genes involved in the proliferation and development of hematopoiesis to lymphatic formation. Thereby GATA2 dysfunction comes along with combined monocyte, dendritic cell, B- and natural

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killer lymphoid cell deficiency accompanied by increased susceptibility for mycobacterial, fungal and viral infections. Beside the eponymous monocytopenia and mycobacterial infections, patients have a high risk of developing mainly viral infection triggered malignancies and hematopoietic disorders such as hypoplastic myelodysplastic syndrome and acute myeloid leukemia. Case: Here we describe the case of a juvenile patient with refractory cytopenia with multilineage dysplasia (RCMD) caused by MonoMAC syndrome who underwent allogeneic hematopoietic stem cell transplantation (HSCT) after prolonged history of severe infection complications including ongoing disseminated Mycobacterium abscessus infections. After initial blaze of the inflammation in response to immune reconstitution, the immune system of the patient recovered sufficiently resulting in a significant improvement of the mycobacterial infection. Bone marrow analyses 18-month posttransplantation showed a complete donor chimerism and a normalization of the hematopoiesis. Besides microbiological testing, the infection surveillance was carried out efficiently by 18F-FDG PET-CT scans. Conclusion: Although the MonoMAC syndrome is a rare disease, severe or recurrent nontuberculous mycobacterial (NTM) and disseminated viral or fungal infections respectively during early adulthood combined with hematological abnormalities should lead to the consideration of a possible MonoMAC syndrome. This case shows that HSCT is an appropriate tool in the treatment of MonoMAC syndrome particularly with regard to the high incidence of myelodysplasia/acute myeloid leukemia in this patient collective. Disclosure: No conflict of interest disclosed. P609

Cerebral toxoplasmosis and probable invasive mucormycosis in an immunocompromised patient after hematopoietic stem cell transplantation Brueckner F.1, Freitag S.1, Wittke C.1, Lakner J.1, Große-Thie C.1, Henze L.1, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany 1

Introduction: Cerebral toxoplasmosis (CT) and invasive fungal disease (IFD) are rare opportunistic infections affecting patients undergoing hematopoietic stem cell transplantation (HSCT). Risk factors beneath individual patient-related are long-term depletion of leukocytes. We present a case of a 46-year old hodgkin’s lymphoma (HL) patient after allogenic HSCT affected by CT and probable IFD namely rhinocerebral mucormycosis. Case report: Our patient was initially diagnosed with HL in 2013, relapsed in 2014 and treated according corresponding standard recommendations. He suffered a second relapse in 2015 treated with brentuximab vedotin, a second autologous HSCT followed by an allogenic matched unrelated HSCT. Conditioning consisted of 12 Gy total body irritation, cyclophosphamide and antithymoglobuline. Graft versus host disease (GVHD) prophylaxis consisted of methotrexate and ciclosporin. Early posttransplant course was unremarkable. On day +113 the patient developed headache and paresthesia in his lower limbs. MRI revealed a cerebral lesion in his left basal ganglia including lowering of the three main metabolites choline, creatinine and especially N-acetylaspartate. Blood and liquor analyses remained negative leading to stereotactic biopsy from the left basal ganglia. DNA analysis revealed toxoplasma gondii infection which was treated with pyrimethamine and clindamycin. Immunosuppressive therapy was stopped. Subsequent MRIs showed a slow size reduction of the lesion including the perifocal edema. In addition to the cerebral lesion the patient developed a cheek swelling and a buccal ulcerative lesion on day +127, i.e. 14 after MRI diagnosis. Multiple microbiological and histopathological examinations remained negative. Clinical diagnosis of a probable rhinocerebral mucormycosis was made and liposomal amphotericin b was started. After 3 weeks of treatment the ulceration and swelling completely disappeared.

Abstracts

Conclusion: Opportunistic infections are challenging for the medical team. Despite modern techniques such as PCR and histopathological examination suspected clinical diagnosis might have to guide treatment. Next to application of broad antimicrobial agents urgent stop of immunosuppression remains a cornerstone of successful treatment. Disclosure: No conflict of interest disclosed. P610

Severe Gemcitabine associated vasculitis – a rare but fatal side effect Schmidt S.1, Henze L.1, Herlyn P.2, Wichelhaus A.2, Mittlmeier T.2, GroßeThie C.1, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany, 2Trauma-, Hand and Reconstructive Surgery, Rostock University Medical Center, Rostock, Germany 1

Introduction: Gemcitabine is a commonly used substance in systemic treatment for several solid cancers. It is usually administered in an intravenous pattern and rarely causes severe complications. Herein, we report a rare case of a patient who developed a rapid onset of Gemcitabine-related vasculitis in all of his digits with fatal outcome. Case report: The 77-year old male was diagnosed with metastatic carcinoma of unknown primary. Treatment with Cisplatin 50mg/m2 i.v. and Gemcitabine 1000mg/m2 i.v. (day 1 and 15, q3w) was started in palliative intend and was well tolerated. On day 18 of the first cycle the patient started to suffer from severe pain, coldness and paresthesia in all of the fingertips. Thus there was a suspicion of a vasculitis with Raynaud`s symptomatic caused by a paraneoplastic syndrome. As differential diagnosis a Gemcitabine induced vasculitis was considered. Therapy was continued with another identic cycle of Gemcitabine/Cisplatin with the intention of treating vasculitis as a paraneoplastic syndrome aiming at full recovery. However, symptoms of vasculitis worsened and physical examination revealed ischemic necrosis in all of the fingertips. Treatment of vasculitis was initiated with high-dose steroids (1mg Prednisolone/kg body weight), antibiotics and analgesics. Duplex sonography of the arteries showed no signs of stenosis, rheumatic diagnostics were negative. After negative steroid response Gemcitabine-associated severe vasculitis with ischemic necrosis was diagnosed. Paraneoplastic vasculitis was excluded, because the CT scan showed partial response after two complete cycles of Gemcitabine/Cisplatin. Treatment options with nitrates or bilateral stellate ganglion block were discussed. We decided to continue treatment with analgesia and antibiotics, as the ischemic necrosis was progressing rapidly. Because of his impairment the patient decided to have his fingers amputated. After the procedure was performed the patient recovered quickly, but underwent surgery again when an infection in one of his digits occurred. Therefore, chemotherapy has not been continued so far. Conclusion: This case illustrates the potential for severe vasculitis when using Gemcitabine and the importance of treatment discontinuation immediately when early signs of vasculitis occur, although the ischemic necrosis seen in this patient is rare. Recognition of this syndrome is critical to the provision of early treatment and prevention of ischemic necrosis. Disclosure: No conflict of interest disclosed. P611

Long-term survival after a tandem autologous and a consecutive allogeneic peripheral blood stem cell transplantation in refractory Burkitt´s lymphoma Klink A.1, Mügge L.-O.1, Schilling K.1, Schmidt V.1,2, Eigendorff E.1, Hilgendorf I.1, Hochhaus A.1, Sayer H.G.1,2 Jena University Hospital, Department of Internal Medicine II, Hematology and Oncology, Jena, Germany, 2HELIOS Klinikum Erfurt, Department of Internal Medicine IV, Hematology and Oncology, Erfurt, Germany 1

Introduction: Burkitt lymphoma (BL) is a highly aggressive non-Hodgkin`s lymphoma and requires primary intensive chemotherapy. 75–90%

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of adults reach complete remission, but relapse or refractory disease have a very poor long-term outcome. We report a case with therapy resistant BL in a 26-year old female patient. Patient and methods: Initially the patient presented with meningeosis leucemica and mediastinal, mesenteric and retroperitoneal lymphoma manifestations (07/2007). She was treated according the GMALL-B-ALL/ NHL 2002 protocol. After two blocks of therapy she achieved partial remission (PR). However, progressive disease occurred during the fifth course of chemotherapy. A salvage therapy with R-DHAP with only brief response was followed by myeloablative conditioning with BEAM and subsequent autologous peripheral blood stem cell transplantation (auto-SCT). Within four weeks she suffered again from progressive disease and infield right cervical irradiation was performed with 12 Gy prior to the scheduled allogeneic SCT (allo-SCT). Due to cancellation of the stem cell donor, cytoreductive therapy with vincristine, prednisolone, cytosine arabinoside and a second myeloablative chemotherapy after conditioning with TEAM (thiotepa, etoposide, cytarabine melphalan) with auto-SCT were performed. Only 22 days later she received unrelated allo-SCT (HLA-10/10 match) after reduced intensity conditioning with tresosulfan 3 x 8 mg/m², fludarabine 5 x 30 mg/m² and rabbit ATG 3 x 10 mg/m²in PR of BL in 04/2008. GVHD prophylaxis was performed with CsA alone. Results: On day +30 after allo-SCT significantly regressive lymphoma structures cervical, mediastinal and abdominally were noted. On day +94 she developed cutaneous acute GVHD (grade II) which was treated with systemic prednisolone and topical corticosteroids. Seven months after allo-SCT complete remission (CR) could be demonstrated. Eight years after allo-SCT, she is still alive with sustained CR of BL. A mild chronic cutaneous GVHD is ongoing, but has no significantly impact on her quality of life. The patient was able to successfully complete her university education and works full-time. Conclusion: This case demonstrates the value of intensive high-dose chemotherapy in controlling proliferative BL before allo-SCT. Long-term follow up of our patient underlines the curative immunotherapy treatment option from the allogeneic stem cells via graft-versus-lymphoma effect. Disclosure: No conflict of interest disclosed. P612

Extensive Russell bodies in a case of light chain multiple myeloma Hastka J.1, Schmutz M.1, La Meir F.1, Hofmann W.-K.1, Metzgeroth G.1 III. Medizinische Klinik, Hämatologie und internistische Onkologie, Universitätsmedizin Mannheim, Mannheim, Germany 1

Introduction: Russell bodies were first described by the pathologist William Russell in 1890, who thought that he had discovered the organism that caused cancer. Abundant Russell bodies within plasma cells define Mott cells, which are named after Frederick Walker Mott, who identified such cells in brains of monkeys with trapanosomiasis. Dutcher bodies, first described in patients with Waldenström macroglobulinaemia, are inclusions that invaginate into or overlay the nucleus. Meanwhile we learned that all these terms describe immunoglobulin inclusions derived from an excessive immunoglobulin overproduction. Methods: A 56-year-old woman was admitted to hospital because of an impaired general performance status, suffering from fatigue, night sweats and weight loss. Laboratory findings revealed anemia of 87 g/L and an acute renal insufficiency with a serum creatinine of 1.8 mg/dL. Immunofixation detected monoclonal IgG-lambda paraprotein in serum and urine. The urine protein excretion was excessively elevated to 9200 mg/ day. Free lambda light chains in serum were increased to 8590 mg/L. Magnetic resonance imaging revealed bone lesions in femur and pelvis. Results: May-Grünwald-Giemsa stained bone marrow smear showed a nearly complete infiltration by atypical, polymorphic plasma cells. About 75% of the latter contained spherical, cytoplasmic basophilic inclusions known as Russell bodies. In some of the plasma cells, the inclusions completely occupied the cytoplasm with an appearance of Mott cells. In numerous cells the globuli overlay the nucleus giving rise to the appearance

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of intranuclear inclusions, formerly referred to as Dutcher bodies. FISH analysis identified an unfavourable karyotype with a del13q14 in conjunction with a del17p13.1 and t(4;14). The patient was treated with lenalidomide-doxorubicin-dexamethasone induction and autologous stem cell transplantation followed by allogeneic transplantation. 12 months later the patient died on infectious complication while in complete haematological and cytogenetic remission. Conclusion: Russell bodies and Dutcher bodies are cytoplasmic inclusions, observed in plasma cells undergoing excessive synthesis of immunoglobulin. The immunoglobulin-containing globuli are embedded within the endoplasmic reticulum. These abnormalities in plasma cells are not specific for malignant disorders, e.g. multiple myeloma, lymphoma but are also seen in chronic inflammation, e.g. tuberculosis, rheumatoid arthritis. Disclosure: No conflict of interest disclosed. P613

Invasive Candida albicans infection with central nervous system manifestation in a patient with acute myeloid leukemia Galfetti E.1, Bögeholz J.1, Grassinger J.1, Müller A.M.1, Mueller N.J.2, Rüegg C.2, Kalias S.3, Schanz U.1 Universitätsspital Zürich, Division of Hematology, Zürich, Switzerland, Division of Infectious Diseases and Hospital Epidemiology, Zürich, Switzerland, 3 Universitätsspital Zürich, Division of Neuroradiology, Zürich, Switzerland 1 2

Introduction: Invasive fungal infection (IFI) remains an important cause of morbidity and mortality in patients with acute myeloid leukemia (AML). In these patients Candida spp is one of the most frequent pathogens causing IFI and the central nervous system (CNS) may rarely be involved. Candida infection can lead to different manifestations in the CNS such as abscess, meningitis and vascular changes. Neuroimaging mostly shows small and multiple micro abscesses. But macro abscess as CNS-Candida manifestation has also been previously described. Case: We present a case of a patient with newly diagnosed AML and initial probable pulmonary fungal infection, showing a rapid clinical evolution to invasive candidiasis with possible involvement of the CNS. Twenty days after starting the first induction chemotherapy, a new clinical manifestation of visual loss was investigated with a MRI of the brain which showed an intra cerebral abscess in the right parietal-occipital region. A cardio embolic etiology was ruled out by TEE. Further diagnostic imaging identified abscesses in the liver, spleen and kidneys as well as intramuscular abscesses in the left forearm and left lower leg. Based on the detection of Candida albicans in the biopsy performed on the abscess of the forearm, the radiologic pattern typical for disseminated candidiasis in the abdomen CT, we presumed that the CNS lesion was a manifestation of the same fungal pathogen. The antifungal therapy was extended to caspofungin and voriconazol with regression of the intraabdominal abscesses and no progression of the intracranial lesion. Conclusion: The prevalence of possible neurocandidiasis in patients with AML may be underestimated. A CNS involvement should be investigated at the presence of clinical features such as encephalopathy or focal neurological signs. Pathological and radiological manifestations are variable and may include micro abscesses, macro abscesses, meningitis and vascular complications. Due to the most common manifestation with micro abscess a MRI is preferred to a CT scan. A rapid and adequate diagnostic should be performed in these high risk patients by clinical presumptions of invasive candidiasis, with emphasis on CNS manifestation which has an important implication for therapy and prognosis. Disclosure: No conflict of interest disclosed.

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P614

Small cell neuroendocrine carcinoma of the right eye: A case report Hoya V.1, Stuschke M.2, Sauerwein W.2, Bornfeld N.3, Metz K.4, Schuler M.1, Kasper S.1 Universitätsklinikum Essen, Westdeutsches Tumorzentrum, Innere Klinik (Tumorforschung), Essen, Germany, 2Universitätsklinikum Essen, Westdeutsches Tumorzentrum, Klinik für Strahlentherapie, Essen, Germany, 3 Universitätsklinikum Essen, Westdeutsches Tumorzentrum, Klinik für Augenheilkunde, Essen, Germany, 4Universitätsklinikum Essen, Westdeutsches Tumorzentrum, Institut für Pathologie, Essen, Germany 1

Introduction: We present the first documented case to our knowledge of a neuroendocrine carcinoma of the right vitreous body. After multidisciplinary treatment with cisplatinum/etoposide based chemotherapy and radiation the patient reached visual improvement without relapse so far. Methods: A 46-year-old man with a past medical history of arterial hypertension and diabetes, working as a craftsman, positive family history for prostate carcinoma (father) and positive nicotine abuse, suffered from a progressive loss of vision of the right eye with left recognition of hand movement. Initial diagnosis was an inflammation of the right visual nerve and subsequently a high-dose steroid-therapy was initiated. Further ophthalmological examination was performed and uncovered a relative afferent pupillary defect and a prominent visual nerve with intraocular bleeding signs. A cranial magnetic resonance imaging (MRI) was without pathological finding. Finally a right eye vitrectomy was performed and a biopsy of the vitreous body was taken because of progressive loss of vision. The final pathologic examination showed a poorly differentiated small cell neuroendocrine carcinoma with positive immunostaining for CD56, CK18, synaptophysin, chromogranin and TTF-1 and a high proliferative index Ki67 of 100%. A whole body computertomographic scan and FDGPET scan, gastroscopy and coloscopy, bronchoscopy, thyroid sonography, lumbar puncture and bone marrow puncture revealed no further malignant manifestation or primary tumor. Results: After discussion in the multidisciplinary tumor board the patient received four cycles of chemotherapy with cisplatinum (50mg/m2 d1+d5) and etoposide (100mg/m2 d2-d4, q3w), followed by a local radiotherapy of the right orbital cavity and a prophylactic whole brain radiation. Vision of the right eye improved during chemotherapy and kept stable after treatment. The last follow-up six months after multidisciplinary treatment showed a complete remission. Conclusion: Small cell carcinoma is a type of highly malignant cancer that most commonly arises within the lung. Very rarely, the primary site for small-cell carcinoma is outside of the respiratory tract, mostly in the gastrointestinal tract. This is the first described case of a small cell carcinoma primary located in the right eye, which was successfully treated with combined radio-chemotherapy. Disclosure: No conflict of interest disclosed. P615

Ebstein-Barr-Virus (EBV) infection induced Pseudothrombocytopenia Austein T.1, Seevers R.1, Baumgardt H.2 St. Bernhard Hospital, Medizinische Klinik II-Abteilung für Hämatologie/ intern. Onkologie, Brake, Germany, 2Praxis für Allgemeinmedizin, Nordenham, Germany 1

examination were regular and there were no signs of neck stiffness, oedema or petechiae. An analysis of the blood count revealed a leukocytosis, severe thrombocytopenia, elevated liver enzymes and a notable increased lactat-dehydrogenase (table 1). Tab. 1. Selected laboratory parameter ad admission

Parameter Leucocytes Hemoglobin Thrombocytes Alanineaminotransferase Gammaglutamyltransferase Lactat-dehydrogenase

Reference lab 4.0-10-0 (*1000/µl) 12-16 (g/dl) 150-400 (*1000/µl)

Result 15,1 13,5 12

0-35 (U/l)

168

0-35 (U/l)

100-247 (U/l)

971

An immediately prepared peripheral blood smear (figure 1) exposed a lot of thrombocytes cluster (black arrow) and many reactive lymphocytes (white arrow) suitable for an acute Ebstein-Barr-Virus (EBV) infection, which was confirmed by following antibody assay.

Fig. 1. Blood smear (Pappenheim staining 1000x).

The black arrow point at a thrombocytes cluster and the white indicate a lymphoblastic lymphocyte. Under symptomatic therapy the patient recovered completely. Two weeks after infection of EBV the blood count was normalized and the pseudo-thrombocytopenia disappeared. Conclusion: An EBV-induced Pseudo-thrombocytopenia is an extremely rare symptom and mostly unknown. To avoid false therapy decision (e.g. thrombocyte infusion or cortisone application), it´s necessary to refer about this phenomenon. Reference: 1 A. T. Hsieh, T. Y. Chao, and Y. C. Chen (2003): Pseudothrombocytopenia Associated With Infectious Mononucleosis. Archives of Pathology & Laboratory Medicine: January 2003, Vol. 127,No. 1,pp:e17-e18. Disclosure: No conflict of interest disclosed.

Introduction: Ebstein-Barr Virus infection is typical in young adults. Usually the infection is self-limiting and complication e.g. splenic rupture, thrombocytopenia are seldom. A severe thrombocytopenia while EBV-infection is well known, but EBV induced Pseudo-thrombocytopenia is extremely rare [1]. Here we told about this phenomenon. Case report: An until now healthy 28-eight-year old female was admitted to the emergency room in course of fever and sore throat. There was no history of pre-existing illness.The clinical investigation showed a pharyngitis and a cervical lymphadenopathy. The heart, lung and abdominal

Abstracts

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P616

A case of primary myelofibrosis achieving transfusion independency with palliative megestrol acetate therapy Sreter L.1, Takats A.2 Endomedix Ambulance, Onkohaematology, Budapest, Hungary, 2Endomedix Ambulance, Internal medicine, Budapest, Hungary 1

Introduction: In the curative therapy of OMF for younger patients with advanced disease, allogeneic stem cell transplantation may be beneficial. Nonmyeloablative allogeneic stem cell transplantation has been successfully used even in older patients. Inhibitors of the JAK pathway appear to have a significant effect on splenomegaly and symptoms; ruxolitinib is currently available and others are being tested in clinical trials. JAK inhibitors are effective whether or not a JAK2 mutation is present. Routinely the therapy is directed at symptoms and complications. Androgens, splenectomy, corticosteroids, chemotherapy, and splenic embolization and radiation therapy are used for palliation. Objectives: The 66 years old patient has primary myelofibrosis since 8 years. The diagnosis was based on bone marrow core biopsy. He was on hydroxyurea therapy (1×500mg/day) for 4 years, than on interferon-a (3×3M NE/week sc. for 6 months. The IFN-a therapy was stopped because of severe thrombocytopenia and anemia. Darbopoietin was given every 3rd week at a dose of 500 mcg sc. Darbopoietin didn’t improve his erythrocyte count. We observed the effect of Darbopoietin for 3 months, than Epoetin beta, 30,000 IU per week in three divided doses was administered sc. for 12 weeks without any improvement. Low dose steroids were also given to symptomatic relief of abdominal pain. In 2014 the enlargement of the spleen increased and he got splenic irradiation (4,5Gy total). Between 2013 and 2015 he needed about 2 units of erythrocyte transfusion at every third week because of severe and symptomatic anemia. In February of 2015 we started to give him megestrol acetate in a dose of 800 mg/day because of fatigue and weight loss. After 3 weeks of this palliative therapy his erythrocyte count rised and slowly became normal. Since that time he doesn’t need any transfusions. His general performance is excellent, he is working 8 hours a day. His thrombocytopenia is unchanged, but he has no bleeding complications. Conclusion: Megestrole acetate has several impacts on oncology patients: impovement of the global quality of life, improvement of appetite and survival advantage in 30%. The most important adverse affect is thrombosis in 30% of the patients. Our patient became transfusion independent – as an unforseen and till now not registered effect of megestrol acetate (17α-acetoxy-6-dehydro -6-methylprogesterone). Disclosure: No conflict of interest disclosed. P617

Atypical clinical course with thromboembolic complications of an patient with acute promyelocyte leukemia Hiemer S.1, Nelk I.1, Dahlenburg J.2, Ohly A.2, Westphal S.3, Plauth M.4, Florschütz A.1 Städtisches Klinikum Dessau, Innere Medizin/ Hämatologie/Onkologie, Dessau, Germany, 2Städtisches Klinikum Dessau, Innere Medizin/ Angiologie, Dessau, Germany, 3Städtisches Klinikum Dessau, Labor, Dessau, Germany, 4Städtisches Klinikum Dessau, Innere Medizin, Dessau, Germany 1

Introduction: In February 2016 we saw the 42 year old female patient the first time to diagnose an unclear panzytopenia which already persisted for several weeks. The patient initially was in hospital because of recent thromboses and embolisms under therapeutical anticoagulation. Methods: To clearify the panzytopenia we made a bone marrow aspiration and we found atypical hypergranulated myeloblasts in zytology with about 20% infiltration. In the flow cytometry the cell population was negative for CD 34 and HLA-DR and positive for CD 117, 38, 33, 13 and POX. Suspecting an atypical APL, we started a therapy with ATRA. We saved the diagnosis by the evidence of t(15;17)(q22;q21) and PML/RARA rearrangement. Because she was a low risk patient (WBC 2.1 Gpt/l; thrombocytes 100 Gpt/l), we initiated an induction therapy with ATO. After two

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weeks of therapy the hemoglobine and the thrombocytes normalised, but the WBC has been increasing every day. Additionally the patients weight increased and she got edemas and dyspnoe. That was the reason to start a high dose therapy with dexamethasone due to a leucocyte differential syndrome. There was neither clinical improvement nor hematological normalisation. Because the leucocytes were still increasing we decided to initiate a classical 3+7 induction chemotherapy. Results: The clinical conditions were regredient and after a chemotherapy induced pancytopenia we saw a normal regeneration of the blood count. We did another bone marrow aspiration on day 15 after starting chemotherapy and saw a complete remission of the blasts. On March 2016 we started the conditional therapy with ATRA and ATO. Conclusion: It has been interesting, that this patient occured to thromboses and embolisms and not to bleeding complications. The morphology of the blasts has been atypical, but we saw typical constellations in the flow cytometry wich was the reason to suspect an APL and start the therapy with ATRA. In the further clinical course the therapy has been well tolerated by the patient and until now we are observing a complete hematological and molecular remission. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium

Biologie der chronischen myeloischen Leukämie V619

The role of the immune system for the achievement and maintenance of deep remission in CML Wolf D.1 Universitätsklinikum Bonn, Medizinische Klinik 3; Onkologie, Hämatologie, Immunonkologie und Rheumatologie, Bonn, Germany 1

The main treatment goal in CML is the achievement of a stable deep molecular response (DMR), in which disease progression is prevented and discontinuation of TKI therapy can be considered. Second generation TKIs induce a higher probability to achieve this goal when compared to imatinib. Since decades, researchers tried to hijack the immune-system to induce improved response rates in CML. Most trials evaluated vaccination strategies aiming at inducing BCR-ABL or leukemia-associated antigen (LAA) specific T cell responses. These active immunotherapy trials however had only limited clinical efficacy. The most promising immuno-activating agents tested in combination with TKIs is recombinant (pegylated) interferon-alpha (IFN-a), which in combination with TKIs induces superior response rates compared to imatinib therapy alone. Other valid clinical data using immo-therapy to improve remission rates are lacking. However, several lines of evidence from patients reaching and maintaining DMR upon TKI discontinuation suggest an important contribution of immune cells in CML control. IFN-a may induce long-term disease control upon disontinuation, which is linked to a specific pattern of CML-reactive CD8+ T cell clones, leading to future randomized testing of IFN-a versus placebo upon TKI disconituation. Moreover, correlative data from the EURO-SKI trial suggest that both, innate and adaptive immune cells contribute to CML-control upon TKI withdrawl, as increased NK cells and low CD86 positive plasmacytoid DC (pDC) are linked to an increased likelihood to maintain remission without TKI. Of note, high CD86+ pDC are linked to an exhausted T cell phenotype (i.e. PD-1 positive LAA-speific CD8 T cells), also suggesting that PD-1 is an attractive target for CML therapy. Indeed, a phase 1/2 clinical trial evaluates the safety and efficacy of combined PD-1 targeting by nivolumab with dasatinib. This is of particular interest, as various recent reports demonstrated immune-modulatory effects of dasatinib leading to LGL lymphocytosis, which is correlated to superior clinical outcome. Thus, the immunological safety of dasatinib with nivolumab is of importance for further development of this combination therapy. In summary, various immune-activating approaches are currently exploited to improve the depth and maintenance of DMR in

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CONTENTS AUTHOR INDEX

CML. However a deeper mechanistic understanding of immune-mediated MRD control is needed to further improve immunotherapy in CML. Disclosure: Dominik Wolf: Financing of Scientific Research: BMS, Novartis, Ariad, Pfizer; Expert Testimony: BMS, Novartis, Ariad, Pfizer V620

CML stem cell biology Schemionek M.1 Institut für Hämatologie, Onkologie, Hämostaseologie und Stammzelltransplantation, Aachen, Germany 1

Chronic myeloid leukemia (CML) is genetically defined by the acquisition of the t(9;22) translocation within the hematopoietic stem cell (HSC) compartment. The derived Bcr-Abl oncoprotein lacks autoinhibitory function and gains the Bcr oligomerization domain, both resulting in the constitutive activation of the Abl tyrosine kinase. The early insight into the molecular pathogenesis of CML paved the way for the implementation of a targeted cancer therapy using tyrosine kinase inhibitors (TKIs) and this has significantly increased therapy outcome. However, already early after TKI therapy start several lines of evidence suggested that a small subpopulation within the CD34+ cell compartment persists despite treatment. Subsequently, the utilization of primary human stem cells and mouse models validated persisting disease-initiating cells despite Bcr-Abl targeting. In line with these reports, the STIM trial has first demonstrated relapse upon therapy discontinuation, in a patient majority, even in the context of deep molecular response. Alluding to the stem cell specific characteristics of self-renew and differentiation potential, this disease-initiating cell population is specified as leukemic stem cells (LSCs). In chronic phase (CP) CML, LSCs are included within a population showing a phenotypic marker expression pattern of CD34+CD38-CD90+Lin-. It has been shown that TKIs maintain their efficacy against the Bcr-Abl tyrosine kinase in this CP-CML LSC compartment and therefore the mechanisms that support their Bcr-Abl kinase independent survival are in the focus of current research efforts that will be discussed here. These studies have tried to uncover critical information on the resistance mechanisms at the cellular (cell-intrinsic) or microenvironmental (cell-extrinsic) level. The variety of described altered signaling pathways in LSCs includes e.g. Wnt/β-Catenin, JAK-STAT, Hedgehog Pathways, ALOX5, PP2A, FOXO/ TGFβ, PI3K/AKT/mTor, Autophagy or CXCR4/CXCL12 activity. However, although our knowledge of LSC biology has increased, we still have not achieved the ultimate objective: The identification of critical and LSC specific pathways that are regulated by druggable targets would provide a curative strategy for CML patients that would free them from the burden of a lifelong pharmacological treatment with a risk of disease relapse. Disclosure: No conflict of interest disclosed.

disease (mAA) or patients without a suitable bone marrow donor should receive immunosuppression (IS) as first-line treatment. For patients with vSAA/SAA the standard regime is horse antithymocyte globuline (hATG) and cyclosporine A (CsA). There are several shortcomings of this gold standard: i) response rates are in the order of 70% and it takes 3–4 months (median) to response, ii) the risk of relapse is up to 40% during long-term follow up, iii) only a subgroup of responding patients achieves complete response. Several attempts have been made to improve response rate, speed of hematopoietic recovery and durability of response by combining hATG/CsA with additional immunomodulators (mycophenolatmofetil, sirolimus) or various hematopoietic growth factors, in particular G-CSF. However, all these attempts did not result in significant superior results in terms of trilineage response rate, relapse rate or survival compared to hATG /CsA. The thrombopoietin-receptor agonist eltrompag is a promising candidate to overcome these shortcomings. It has been demonstrated that it promotes hematopoiesis by stimulating hematopoietic stem and progenitor cells. A clinical trial in patients with refractory SAA achieved an overall response rate of 40% to eltrombopag. Another trial evaluated the addition of eltrombopag to hATG / CsA in treatment-naïve (v)SAA patients. This increased the overall response rates and the complete response rates. An increase of CD34+ cell, appearance of multipotent progenitor cells and rapid recovery of peripheral blood counts was observed. In the meantime two prospective controlled clinical trials have been launched: i) RACE trial, a randomized open label trial comparing hATG/ CsA vs. hATG/CsA eltrombopag in patients with vSAA/SAA who are not eligible for allogeneic stem cell transplantation. ii) EMAA trial, a randomized placebo-controlled trial comparing CsA+placebo vs. CsA+eltrombopag in patients with mAA. We will discuss mechanism of action, current results of clinical trials, potential risks (clonal evolution) and design of new trials with thrombopoietin-receptor agonists for treatment of AA. Disclosure: Hubert Schrezenmeier: Advisory Role: Novartis; Financing of Scientific Research: Novartis; Expert Testimony: Novartis Britta Höchsmann: Advisory Role: Novartis; Financing of Scientific Research: Novartis; Expert Testimony: Novartis

Fortbildung

Blutung – Gemeinsames Symposium mit der Gesellschaft für Thrombose- und Hämostaseforschung e. V. (GTH) V627

Congenital and acquired hemophilia: update 2016 Tiede A.1 Hämatologie, Hämostaseologie, Onkologie und Stammzelltransplantation, Hannover, Germany 1

Wissenschaftliches Symposium

Acquired aplastic anemia (AA) is a rare disorder which is characterized by bicytopenia or pancytopenia and hypoplastic bone marrow. Allogeneic bone marrow transplantation is the treatment of choice for young patients (<40–50 years) with severe AA (SAA) or very severe AA (vSAA) and an HLA-identical sibling donor. Older patients, patients with non-severe

Congenital haemophilia A and B results from inherited mutation of factor VIII (FVIII) or factor IX (FIX) genes, respectively. In contrast, acquired haemophilia results from neutralising autoantibodies, almost always directed against FVIII. Severe congenital haemophilia is commonly treated by regular prophylactic infusion of FVIII or FIX, thereby reducing bleeding into joints by about 90% and preventing haemophilic arthropathy. Because of the short half-life of FVIII (12 h) and FIX (18 h), frequent intravenous injection is required to achieve effective factor activity. Recombinant factor concentrates have recently been developed with 1.5fold (FVIII) or even 5fold (FIX) prolonged half-life due to Fc or albumin fusion technology or polyethylene glycol attachment. Novel non-replacement therapies are currently in phase 3 clinical trials, including FVIIIa mimetic antibody (ACE910, emicizumab), an inhibitory antibody against tissue factor pathway inhibitor (mAb 2021, concizumab), and small interfering RNA to suppress antithrombin biosynthesis (ALN-AT3, fitusiran). Potential advantages of these therapies include efficacy in the presence of neutralising antibodies against FVIII or FIX, subcutaneous application and long half-life. A porcine sequence, recombinant FVIII (susoctocog

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–192

Zytopenien V625

TPO agonists and bone marrow failure Schrezenmeier H.1, Körper S.2, Höchsmann B.2 Institut für Transfusionsmedizin, Universität Ulm, Institut für Klinische Transfusionsmedizin und Immungenetik Ulm, DRK-Blutspendedienst Baden-Württemberg – Hessen und Universitätsklinikum Ulm, Ulm, Germany, 2 Institut für Transfusionsmedizin, Universität Ulm, Institut für Klinische Transfusionsmedizin und Immungenetik Ulm, DRK-Blutspendedienst BadenWürttemberg – Hessen, Ulm, Germany 1

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alfa) was recently developed that can rescue haemostasis in the presence of autoantibodies against FVIII because of low or absent of cross-reactivity. The drug has recently been licensed for treatment of bleeds in acquired haemophilia. Long-term goal in acquired haemophilia is, however, the elimination of autoantibodies against FVIII by immunosuppressive therapy (IST). A study of the German, Austria and Swiss Society on Thrombosis and Haemostasis (GTH) studied efficacy and safety of a standardised IST protocol and described prognostic factors for remission and survival of acquired haemophilia. Low FVIII activity at the time of first diagnosis was associated with longer time to partial remission (FVIII < 1%, 43 days; FVIII >=1%, 24 days). Of note, FVIII autoantibodies of the IgA class were a risk factor for early recurrence (IgA negative, 12%; IgA >1:80, 50%). Complete remission after cessation of IST was maintained by 70% of IgA negative, but only 8% of IgA >1:80 patients. These prognostic factors may help to better tailor IST in future studies. Disclosure: Andreas Tiede: Advisory Role: Bayer, Baxter/Baxalta, Biogen Idec, Biotest, Boehringer Ingelheim, CSL Behring, Leo Pharma, Novo Nordisk, Octapharma, Pfizer, SOBI.; Expert Testimony: Bayer, Baxter/Baxalta, Biotest, CSL Behring, Novo Nordisk, Octapharma, Pfizer, SOBI. V628

Management of bleeding emergencies in patients treated with DOACs Jaschonek K.1 Universitätsklinikum Tübingen, Medizinische Klinik und Poliklinik, Onkologie, Hämatologie, Rheumatologie und Pulmologie, Tübingen, Germany 1

DOACs promise convenient fixed dosing with fewer interactions and quicker onset/offset of action compared with VKA. However, prescribing of these compounds remains challenging in daily practice for several reasons. There are multiple dose regimens for different indications and populations (e.g. older patients, patients with renal impairment, or patients on potentially interacting drugs). Indeed, DOACs are often prescribed incorrectly, and off-label use is common. Registry data show that DOACS are not used according to stroke risk scores and anticoagulation is still being widely underused in atrial fibrillation. One reason may be prescribers’ uncertainty when it comes to handling DOAC-associated bleeding. Although the rate of intracranial hemorrhage is low compared with VKA, about 1–4% of the patients receiving DOACs may experience major bleeding or require rapid reversal of anticoagulation for emergency surgery. In the event of life threatening hemorrhage, PCCs, aPCCs and rVIIa have been considered. However, no consensus is available for treatment protocols with these compounds. Monitoring and tailoring of the reversals remain a major issue. Specific reversal agents (antidots) for DOACs are currently under clinical investigation in Phase I/II studies. Tested substances include andexanet alfa (Portola Pharmaceuticals), idarucizumab (Boehringer Ingelheim), and PER977 (aripazine, Perosphere Inc.). Andexanet alfa is a modified, recombinant human fXa molecule (decoy) that retains high-affinity binding to direct fXa inhibitors and is an universal factor Xa inhibitor reversal agent. Idarucizumab is a fully humanized monoclonal antibody fragment (FAb) against dabigatran. PER 977 is a small molecule non-specific reversal agent. Further studies are needed to demonstrate clinical outcome benefits of these new compounds and high-quality postmarketing evaluation has been recommend to monitor their safety and appropriate use. Disclosure: Karl Jaschonek: Advisory Role: CSL Behring; Financing of Scientific Research: CSL-Behring, Bayer, Boehringer-Ingelheim; Immaterial Conflict of Interests: Vorsitzender der Arzneimittelkommission des UKT

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V629

From JAK2 to von WIllebrand: bleeding and thrombosis events in MPNs Koschmieder S.1 Universitätsklinikum der RWTH Aachen, Klinik für Onkologie, Hämatologie und Stammzelltransplantation, Aachen, Germany 1

Myeloproliferative neoplasms (MPNs) are clonal disorders of hematopoietic stem cells and their progeny, and they are characterized by an increase of one or more blood cell lineages in the peripheral blood, bone marrow hyperplasia, and splenomegaly due to extramedullary hematopoiesis. Clinically, patients with MPN are at risk of developing complications such as thromboembolic/thrombotic and severe hemorrhagic events. During this presentation, I will review the incidence, diagnosis, clinical presentation, and treatment of bleeding and thrombosis events in the classical Philadelphia negative MPN subtypes, i.e. polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). In particular, the current standards for molecular diagnosis, (i.e. JAK2V617F, MPL W515K/L, CALR mutations, and exclusion of BCR-ABL transcripts), clinical risk stratification (including risk factors for bleeding and thrombosis), and risk-directed hemostatic and antithrombotic therapy as well as molecularly targeted therapy will be presented. Finally, specific scenarios of bleeding and thromboembolism/thrombosis, such as portal and splanchnic vein thrombosis, acquired von Willebrand syndrome, and perioperative management in MPN will be discussed. Disclosure: Steffen Koschmieder: Advisory Role: Ariad, AOP, Baxalta, CTI, Novartis, Shire, Janssen-Cilag, Bristol-Myers Squibb, Pfizer, Sanofi; Financing of Scientific Research: Ariad, Alexion, AOP, Baxalta, Celgene, CTI, Pfizer, Sanofi, Novartis, Shire, Janssen-Cilag, Bristol-Myers Squibb; Expert Testimony: Novartis, Novartis Foundation, Bristol-Myers Squibb

Wissenschaftliches Symposium

Patentrechtliche Probleme bei der personalisierten Therapie V630

Patents on genes? Perspectives from a diagnostic laboratory Haferlach T.1 MLL Munich Leukemia Laboratory, München, Germany

Genes are important in a state of the art diagnostic approach in hematology. Our knowledge about involved genes, mutations, gains and losses, translocations and other structural changes have dramatically increased over the last 15 years. When detected, several institutes, universities, companies or researchers filed patents. They describe the new finding with respect to the genetically aberration, often combined with techniques to pick up this information at diagnosis and also may use the respective marker for targeted treatment or follow-up in minimal residual disease. If commercial laboratories offer the investigation of the respective gene, they need to know about the filed patents, the respective structures or assays or targets filed and to respect this intellectual property (IP). However, in most instances, the investigation of the respective genes follow home brew assays or individual laboratory approaches and in very few cases tests that are on the market (that include and respect the IP of the founder of the gene) can be purchased. Therefore, routine laboratories that need to be up to date with respect to their portfolio increasingly need to discuss, how they address the medical need to serve patients and doctors and in parallel respect intellectual property. The payment of royalties to the individual patent owner is one option. However, in times of next generation sequencing we already see panel designs including in some cases 50 but also up to 500 genes in some assays already in routine use and accredited in the United States of America. It seems to be very challenging to balance the medical need of patient´s care, patent rights for IP and commercial rules to make any kind of molecular service from a routine laboratory possible as needed for patients care. This is even more true for targeted treatment approaches and the detection of minimal residual disease for

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further treatment strategies. Many aspects in this field so far await answers and the legacy policy is different in US as well as in Europe and depends on the respective country as well. We urgently need some ground breaking decisions like the BRACA1 and BRACA2 decision from the US Supreme Court to allow also routine diagnostic laboratories a state of the art diagnostic approach without being automatically at risk to be sued for their services. Disclosure: Torsten Haferlach: Employment or Leadership Position: MLL Muich Leukemia Laboratory V632

Aspects of patent law for hematologic and oncologic diagnostics Birghan C.1 TBK Patent- und Rechtsanwälte, München, Germany

Numerous technical inventions and discoveries in the field of life sciences during the last decades have revolutionized the understanding of the function or malfunction of the human body at a molecular level, providing new and improved possibilities in hematologic and oncologic diagnostics. In turn, these scientific and technical developments have resulted in a steadily growing number of patents and patent applications directed at inventions in this technical field. The main aim of the worldwide patent system is the promotion of technical and industrial progress in all areas of technology including biotechnology and related fields. However, the enforcement of intellectual property rights including patents in the field of molecular diagnostics may result in practical and ethical problems for the patent proprietors, the diagnostic laboratories as well as the practitioners. The fundamental possibilities and the limits for obtaining patents concerning diagnostic methods involving the application of genes and corresponding methods in molecular diagnostics in the European the US and other relevant international patent systems will be presented. The political and legislative developments in this area as well as the concrete legal basis and the recent case law will be reviewed. Precautionary measures for avoiding patent infringement proceedings and the options for creating proactive strategies for patent seekers and owners, on the one hand, and diagnostic laboratories and practitioners, on the other hand, will be discussed. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium Immunphänotypisierung V634

Automated Approaches for Flow-Cytometric Immunophenotyping Dworzak M.N.1, Reiter M.1, Diem M.2, Kleber F.2, Rota P.2, Karawajew L.3 St. Anna Kinderkrebsforschung, Wien, Austria, 2TU Wien, Computer Vision Lab, Wien, Austria, 3Charité – Universitätsmedizin Berlin, Berlin, Germany 1

Multi-color flow cytometry (FCM) is a mainstay of leukemia diagnostics including assessment of minimal residual disease (MRD). Because of its high prognostic value for risk stratification and treatment planning especially in childhood acute lymphoblastic leukemia (ALL), the FCM-MRD methodology is applied world-wide. However, such assessments are still operator-dependent leading to substantial costs regarding training and quality control. The EU founded AutoFLOW project aims at the standardization and automation of FCM-MRD analysis by machine learning technology. Identifying biologically meaningful cell sub-populations automatically by FCM is essentially a clustering problem, however, standard clustering methods are impractical, because size, shape and location of correspond-

Abstracts

ing clusters may vary strongly between samples mainly due to phenotypic differences and inter-laboratory variations. The basic idea behind the AutoFlow approach is to learn a model from a set of training samples and allow it to adapt to the peculiarities of a new unseen sample in order to perform a more accurate automatic assessment. In the AutoFlow approach, a new input sample is reconstructed by a linear combination of artificial reference samples each represented by a Gaussian Mixture Model (GMM), in which for each Gaussian component the class label of the corresponding cluster of observations is known. The reference samples are calculated from a larger set of training samples by non-negative matrix factorization and can be regarded as the basis of a lower dimensional feature space, in which input samples are reconstructed. The feature space representation of the sample is then used to assign each observation to one of the specified sub-populations by a Bayes decision. Validation data of this approach will be shown, confirming that the model-adaptation is beneficial for this type of application and allows correct automated analysis of samples containing even small amounts of MRD (down to 100 in a million events). Notably, the AutoFlow approach is not limited to the exemplified application. It can be employed wherever analysis of large, multi-dimensional, numerical data of a specific class of samples with related structure has to be performed. Disclosure: No conflict of interest disclosed. V635

Flow cytometric immunophenotyping for the diagnosis of peripheral T/NK cell neoplasia Westermann J.1 Charité – Universitätsmedizin Berlin (CVK), Med.Klinik m.S. Hämatologie, Onkologie und Tumorimmunologie, Berlin, Germany 1

Peripheral T/NK cell neoplasia are a heterogenous group that represents approximately 10–15% of all newly diagnosed lymphoma in western countries. According to WHO 2008, T/NK cell neoplasia can basically be devided into specified and unspecified entities, peripheral T cell lymphoma (PTCL) - not otherwise specified (NOS) and angioimmunoblastic T cell lymphoma (AILD) belong to the most frequent ones. From a clinical point of view, T/NK cell neoplasia can be subdivided into leukemic (i.e. T-PLL, LGL), nodal (i.e. PTCL-NOS, AILD), extranodal and cutaneous (i.e. mycosis fungoides) subforms. Despite modern diagnostic tools including (molecular) histopathology, flow cytometry and molecular genetics, clinical diagnosis of T/NK cell lymphomas remains challenging in many cases. Flow cytometric diagnosis of T/NK cell neoplasia is primarily based on the detection of an aberrant phenotype within a T cell population. These aberrant T cell phenotypes mainly consist of 1) abnormal expression levels of pan T cell markers such as CD3, CD2, CD5, CD7; 2) quantitative or qualitative changes in T cell subsets (CD4+, CD8+, CD4+CD8+ double positive (DP), CD4-CD8- double negative (DN)); 3) expression of additional markers such as CD56, CD57, CD30, CD10, CD25; 4) diminished CD45 expression and 5) aberrant light scatter properties. In our lab, routine diagnostics of (leukemic) T/NK cell lymphoma usually comprise multiparameter 8 colour flow cytometry. The first step in the diagnostic workup is the identification of an aberrant T cell population by the above mentioned criteria and its categorization into four different subsets of T cells: 1) CD4+; 2) CD8+; 3) CD4+CD8 DP and 4) CD4-CD8- DN. Within these different subgroups, the expression pattern of additional markers such as CD10, CD7, CD56, CD57, CD16 and CD30 may help to distinguish between different subgroups of lymphoma. Clonality can either be assessed by using TCR-b-specific antibodies for flow cytometry or by PCR-based techniques for the detection of a TCR gene rearrangement. However, the distinction of T/NK cell lymphoma from reactive changes remains challanging in many cases. Therefore, the integration of flow cytometric results with cytology, (molecular) histopathology and clinical information is crucial. Disclosure: No conflict of interest disclosed.

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V637

Significance of flow cytometric and molecular genetic findings in patients with cytopenias and limited or no signs of myelodysplasia by cytomorphology Kern W.1, Meggendorfer M.1, Fasan A.1, Haferlach C.1, Haferlach T.1 MLL Munich Leukemia Laboratory, Munich, Germany

Introduction: Diagnosis of myelodysplastic syndromes (MDS) remains a challenge in many cases. We assessed the significance of cytomorphologically (CM) borderline dysplastic changes and of flow cytometric (FC) MDS-related findings in the absence of a clear-cut diagnosis of MDS by screening for molecular mutations and by diagnostic reassessment during follow-up (FU). Methods: 322 patients (pts) were assessed for suspected MDS by CM, FC and cytogenetics (CG) which 1) did not have MDS by CM, 2) had normal CG and 3) had ≥1 FU assessment. By CM 159 (49%) pts had borderline dysplastic findings while 163 (51%) had no sign of MDS. By FC 138 (43%) pts were in agreement with MDS according to ELN (≥3 aberrantly expressed antigens, AEA), 141 (44%) had borderline findings (1–2 AEA) and 43 (13%) had no signs of MDS. 699 FU samples were analyzed (median 2/pt). In 147/322 pts (46%) mutational screening was performed on initial samples targeting ASXL1, TET2, RUNX1, SRSF2, BCOR, DNMT3A, IDH2, NPM1, SF3B1, TP53, ZRSR2, CBL, CSF3R, ETV6, KDM6A, KRAS, MLL, SETBP1, SMC3, U2AF1 (median 4 genes/pt, range 1–20). Results: 145 pts (45%) were diagnosed MDS by CM during FU after a median of 3.4 years. More pts with initially borderline dysplastic findings by CM were diagnosed MDS at FU than those without (52% vs 39%, p = 0.025). More pts with initial findings by FC in agreement with MDS were diagnosed MDS at FU than those without (58% vs 35%, p < 0.001). Duration until MDS diagnosis significantly differed between the groups defined by FC and was shortest in pts in agreement with MDS at initial assessment and longest in those without any AEA (median 1.9 vs 5.6 years, p < 0.001). While initial CM results revealed no impact on overall survival (OS), pts with an initial FC result in agreement with MDS had shorter OS (5 year OS 70% vs 88%, p = 0.12). In 21/147 pts (14%) initial molecular screening revealed mutations in ASXL1 (6 pts), TET2 (6), RUNX1 (3), SRSF2 (3), 2 pts each for BCOR, DNMT3A, IDH2, NPM1, SF3B1, TP53 and ZRSR2 and 1 pt each for CBL, CSF3R, ETV6, KDM6A, KRAS, MLL, SETBP1, SMC3 and U2AF1. Significantly more pts with findings in agreement with MDS by FC had ≥1 mutation as compared to those with 1–2 AEA and to those with none (21% vs 10% vs 0%, p = 0.012). Conclusions: Present data argues for an integrated diagnostic approach for suspected MDS including FC together with CM and CG. Implementation also of molecular data on mutations may further improve the validity of MDS diagnostics. Disclosure: Wolfgang Kern: Employment or Leadership Position: Führungsposition Torsten Haferlach: Employment or Leadership Position: Führungsposition V638

Validation of flow cytometric tests in hematology Sack U.1, Dorn-Beineke A.2 University of Leipzig, Medical Faculty, Clinical Immunology and Transfusion Medicine, Leipzig, Germany, 2Helios Dr. Horst Schmidt Kliniken Wiesbaden, Institute of Laboratory Diagnostics and Hygiene, Wiesbaden, Germany 1

Flow cytometric methods are well established in hematological diagnostics, process control, and even preparation of cellular therapeutics. Every of these topics are subject to regulatory systems that are as far as possible harmonized in the European community and worldwide. By growing implementation of ISO 15189 based standardization in European laboratory diagnostics, flow cytometric laboratories are more and more challenged to introduce compliant quality management systems. Alternative quality assurance systems and guidelines request similar affords. So, proof of following regulatory rules is widely requested, namely in preclinical and clinical studies. Quality control in transfusion medicine and ATMP man-

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ufacturing is daily practice. In general, adherence to quality management systems is considered to be very hard in cytometry. Validation processes represent a central challenge in flow cytometric laboratories. Limitations in well defined samples, harmonized analysis processes, and experience in multicolor analysis make validation approaches a challenge for the laboratories. Therefore, we analyzed validation processes and regulatory requests for cytometric labs and investigated flow cytometrists’ attitudes and misgivings according these requirements. As major challenges, staff qualification, adaptation of multicolor antibody panels, and harmonization between various platforms has been identified. References: ISO 15189:2012. Medical Laboratories – particular requirements for quality and competence. Geneva: ISO 2012. Nebe T, Dorn-Beineke A, Braun P, Daniel V, Ilieva Z, Kuling G, Meisel C, Oelschlägel U, Sack U: Messunsicherheit und Qualitätssicherung im Bereich der Immunphänotypisierung der Lymphozytensubpopulationen im peripheren Blut. Laboratoriumsmedizin 37(2013):233–250. Sack U: Zelluläre Diagnostik und Therapie. Berlin/Boston: De Gruyter 2016. Disclosure: Ulrich Sack: Advisory Role: Fachgutachter der DAkkS und von INAB Alexandra Dorn-Beineke: Advisory Role: Fachgutachterin der DAkkS

Wissenschaftliches Symposium

Systemmedizinische Ansätze in der Hämatologie / Onkologie V639

Dynamics and topology of hematopoiesis inferred through genetic labelling of stem cells in situ Höfer T.1, Barile M.1, Busch K.2, Roessler J.1, Schuon A.-K.2, Flossdorf M.1, Pei W.2, Postrach D.2, Feyerabend T.2, Rodewald H.-R.2 Deutsches Krebsforschungszentrum (DKFZ), Abteilung für Theoretische Systembiologie, Heidelberg, Germany, 2Deutsches Krebsforschungszentrum (DKFZ), Abteilung für Zelluläre Immunologie, Heidelberg, Germany 1

The experimental study of hematopoiesis has largely relied on the transplantation of stem and progenitor cells into irradiated hosts and clonogenic assays in vitro. We have developed mathematical approaches for inferring the dynamics of hematopoiesis from unperturbed hematopoietic stem cells (HSCs), collaborating with Hans-Reimer Rodewald and colleagues who have devised novel experimental fate mapping tools for this purpose. Our results reveal fundamental differences between the hematopoietic dynamics in the normal bone marrow and post transplantation in mice. Under steady-state conditions, very many stem cells contribute to differentiated progeny but an individual HSC does so infrequently; by contrast, after transplantation only few engrafted HSC dominate the rebuilding of the system and are highly active. Moreover, our data allow us to determine differentiation and proliferation rates in the hematopoietic stem and progenitor cell compartments, thus yielding a quantitative picture of the cell flux in the myeloid and lymphoid branches of the system. With this combined experimental and theoretical approach, we have begun to study the coordinated responses of hematopoietic stem and progenitor cell compartments to challenges (such as bleeding and irradiation) and, using in situ genetic labeling of individual HSCs, to assess the topology and regulatory linkage of the hematopoietic differentiation tree in situ. Disclosure: No conflict of interest disclosed. V640

In-vivo imaging of normal and leukaemic stem cells Krause D.S.1 Georg-Speyer-Haus und Goethe Universität Frankfurt, Frankfurt am Main, Germany 1

Sophisticated technology now allows real time in-vivo imaging of the normal and haematopoietic stem cell niche providing novel insight into its structure, complexity and dynamics and into how it regulates haemato-

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poiesis. In addition, this talk will focus on the slowly emerging data on the differences between the normal and leukaemic stem cell niches and on differences in interaction with the bone marrow microenvironment even between myeloid leukaemias driven by different oncogenes. Disclosure: Daniela Krause: Advisory Role: Glycomimetics Inc., USA V641

BCR-ABL-Dynamics under TKI-therapy and in therapy-free remission Röder I.1 TU Dresden, Institut für Medizinische Informatik und Biometrie (IMB), Dresden, Germany 1

Introduction: The correct prediction of the proportion of BCR-ABL positive cells in patients with tyrosine-kinase-inhibitor(TKI)-treated chronic myeloid leukemia (CML) has a high clinical relevance. The reason for this is the fact that residual, BCR-ABL positive leukemic (stem) cells can potentially induce relapses or treatment resistances. The BCR-ABL level is usually determined in the peripheral blood of patients using PCR methods. These measurements allow a quantitative description of the treatment response as long as the detection threshold of the PCR is not reached. If BCR-ABL levels fall below the detection limit, the treatment kinetics cannot be described quantitatively anymore. Methods and results: To use the BCR-ABL time courses in the peripheral blood for a quantitative prediction of the long-term response (potentially also after treatment cessation), we propose a computational strategy that combines two modelling approaches. First, we describe the BCR-ABL kinetics by a regression model, which considers both, detectable and un-detectable (i.e. censored) BCR-ABL measurements. Based on this statistical description, we determine optimal parameters of a mechanistic, single cell-based mathematical model of CML, which allows to predict the proportion of residual leukemic cells in the peripheral blood and within the population of hematopoietic stem cells in the bone marrow, based on the peripheral blood measurements. Conclusion: The suggested strategy can be applied to different TKIs and it is suitable to support clinical decision-making using model-based predictions of the treatment response in TKI-treated CML. Disclosure: Ingo Röder: Financing of Scientific Research: Bristol-Myers Squibb; Expert Testimony: Bristol-Myers Squibb V642

Modelling combined immuno-chemotherapy of high-grade non-Hodgkin lymphoma Scholz M.1, Rösch K.1, Hasenclever D.1 IMISE/Universität Leipzig, Leipzig, Germany

Introduction: Moderate intensifications of CHOP-21 chemotherapy improved the outcome of high-grade non-Hodgkin lymphoma in elderly patients, but highly intense therapies are known to be inferior. Adding immunotherapy with Rituximab was a break-through, but levels observed differences in chemotherapy. Ongoing trials attempt to optimize R-CHOP type regimens. To explain these phenomena and to allow predictions of yet untested therapy options, we hypothesise that the immune system has a key role in controlling residual tumour cells after treatment and construct a mechanistic model of therapy outcome thereon. Methods: We propose a differential equations based model of the dynamics and interactions of tumour and immune cells under chemotherapy. Major model features are an exponential tumour growth, a modulation of the production rate of effector cells by the presence of the tumour (immunogenicity) and mutual destruction of tumour and immune cells. Chemotherapy causes damage to both, immune and tumour cells. Immuno-therapy by the monoclonal antibody Rituximab is modelled by a direct cell kill and an intensified immune response. A pharmacokinetic model of Rituximab applications is added. Growth rate, chemosensitivity, immunogenicity and initial size of the tumour are assumed to be patient-specific,

Abstracts

resulting in heterogeneity regarding therapy outcome. Maximum-entropy distributions of these parameters were estimated on the basis of data of randomized clinical trials. Results: The resulting model can explain the outcome of eight different chemotherapeutic regimens with and without Rituximab and corresponding hazard-ratios. Estimated parameters are biologically plausible. The model provides a mechanistic explanation why too intense chemotherapy could be detrimental. We demonstrate how the model can be used to make predictions regarding ongoing clinical trials and yet untested therapy options. We predict for example that current R-CHOP variants for elderly patients have only limited optimization potential. Conclusion: We conclude that our model explains observed effects in lymphoma therapy by the simple assumption of a relevant anti-tumour effect of the immune system. Heterogeneity of therapy outcomes can be traced back to heterogeneity of a few model parameters whose distribution can be estimated on the basis of clinical survival data. The model can be used to predict the performance of new therapy options. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Kolorektale Tumoren V643

FOLFOX plus Bevacizumab (Bev) vs. FOLFOXIRI plus Bev in advanced Colorectal Cancer (CRC): A randomized phase II trial (AIO KRK 0209, CHARTA) Stein A.1, Garlipp B.2, Junghanß C.3, Leithäuser M.4, Vogel A.5, Schaefers M.6, Kaiser U.7, Hoeffkes H.-G.8, Florschütz A.9, Rüssel J.10, Kanzler S.11, Edelmann T.12, Forstbauer H.13, Göhler T.14, Hannig C.15, Hildebrandt B.16, Roll C.17, Steighardt J.18, Meinert F.19, Cygon F.19, Schmoll H.-J.19 Universitätsklinikum Hamburg Eppendorf, Hamburg, Germany, 2Otto von Guericke University Hospital, Magdeburg, Germany, 3University Hospital Rostock, Department for Internal Medicine Clinic III, Hematology, Oncology, Palliative Care, Rostock, Germany, 4University Hospital Rostock, Rostock, Germany, 5Hannover Medical School, Hannover, Germany, 6Onkologische Schwerpunktpraxis, Düsseldorf, Germany, 7St Bernhard Krankenhaus Hildesheim, Hildesheim, Germany, 8Klinikum Fulda gAG, Fulda, Germany, 9 Städtisches Klinikum Dessau, Klinik f Innere Medizin, Hämatologie/Onkologie, Dessau, Germany, 10Martin Luther University Halle-Wittenberg Halle, Department of Oncology and Hematology, Halle, Germany, 11Leopoldina Krankenhaus, Schweinfurt, Germany, 12Schwerpunktpraxis für Hämatologie und Onkologie, Schkeuditz, Germany, 13Praxisnetzwerk Hämatologie und Internistische Onkologie, Troisdorf, Germany, 14Onkozentrum Dresden, Gemeinschaftspraxis Göhler & Dörfel, Dresden, Germany, 15Onkologische Praxis, Bottrop, Germany, 16Charité – Campus Virchow Klinikum (CVK), Medizinische Klinik mit Schwerpunkt Hämatologie und Onkologie, Berlin, Germany, 17Klinikum Magdeburg gGmbH, Klinik für Hämatologie und Onkologie, Magdeburg, Germany, 18Martin-Luther-Universität Halle-Wittenberg, Medizinische Fakultät, Koordinierungszentrum für Klinische Studien (KKS), Halle, Germany, 19Martin Luther University, University Clinic Halle, Halle, Germany 1

Background: FOLFOX or FOLFIRI plus Bevacizumab are standard firstline treatments in advanced CRC. The TRIBE Trial has investigated the role of adding Oxaliplatin to FOLFIRI / Bev. The CHARTA trial investigates the role of adding Irinotecan in addition to FOLFOX / Bev in a randomized Phase II trial. Methods: From 7/11 to 12/14 250 patients have been randomized to standard FOLFOX / Bev vs. FOLFOXIRI / Bev. The schedule of the 4-drug-regimen was identical to the TRIBE trial. In the first 2 cycles, 25% dose reductions have been allowed based on an individual decision in case of impaired performance status, e.g. older age or other individual factors. Inclusion criteria was ECOG 0–2, at least one measurable lesion with size >1 cm, life expectancy > 3 months. Randomisation was stratified according to ESMO Group 1–3 (Schmoll et. al. Ann Oncol 2012) Induction treat-

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ment was given for 6 months, followed by maintenance with Capecitabine/ Bevacizumab until progression or intolerable toxicity, for a maximum of 12 months (total treatment duration 18 months). At progression, the use of reinduction with the initial Oxaliplatin-based combination was due to the discretion of the investigator. Primary endpoint is PFS at 9 months, with COX-model to adjust for group 1–3 - differences. Secondary endpoint is response rate, secondary resection, PFS, OS, separated for group 1–3 and RAS/ or B-RAF status. Results: After a minimum follow-up of 20 months and a maximum follow-up of 58 months, the final results for efficacy and toxicity will be presented including subgroup analysis by ESMO groups, molecular subgroups etc. This is the first presentation of the results and the presented data will be final. The analysis will be finished 08/2016, the data presented at ESMO and the more mature data will be prepared for DGHO. Disclosure: No conflict of interest disclosed. V644

Preoperative chemoradiotherapy and postoperative chemotherapy with capecitabine and oxaliplatin vs. capecitabine alone in locally advanced rectal cancer: final analyses Stein A.1, Schmoll H.-J.2, Hofheinz R.D.3, Price T.J.4, Nordlinger B.5, Daisne J.-F.6, Janssens J.7, Brenner B.8, Schmidt P.9, Reinel H.10, Hollerbach S.11, Caca K.12, Fauth F.W.B.13, Hannig C.14, Zalcberg J.R.15, Marreaud S.16, Tebbutt N.C.17, Mauer M.E.18, Lutz M.P.19, Van Cutsem E.20, Haustermans K.21 Universitätsklinikum Hamburg Eppendorf, Hamburg, Germany, 2Martin Luther University, Division Clinical Oncology, University Hospital, Halle, Germany, 3University Medical Centre Mannheim, Department of Hematology and Medical Oncology, Mannheim, Germany, 4University of Adelaide, Queen Elizabeth Hospital, Adelaide, Australia, 5Hopital Ambroise Pare, Boulogne Billancourt, France, 6Clinique et Maternité Sainte Elisabeth, Namur, Belgium, 7 AZ Turnhout – Campus Sint-Elisabeth, Turnhout, Belgium, 8Rambam Medical Center & Bruce Rappaport Faculty Medicine, Haifa, Israel, 9Private Practice, Neunkirchen, Germany, 10Leopoldina Krankenhaus, Schweinfurt, Germany, 11 Allgemeines Krankenhaus Celle, Celle, Germany, 12Klinikum Ludwigsburg, Ludwigsburg, Germany, 13Onkologische Schwerpunktpraxis, Hanau, Germany, 14 Schwerpunktpraxis für Hämatologie und Onkologie, Bottrop, Germany, 15 Peter MacCallum Cancer Centre, East Melbourne, Australia, 16European Organisation for Research and Treatment of Cancer, Brussels, Belgium, 17 Heidelberg Repatriation Hospital Olivia Newton-John Cancer & Wellness Centre, Heidelberg, Germany, 18EORTC Headquarters, Brussels, Belgium, 19 EORTC, Brussels, Belgium, 20University Hospitals Leuven, Leuven, Belgium, 21 UH Gasthuisberg, Leuven, Belgium 1

Background: The PETACC-6 trial investigates whether the addition of oxaliplatin to preoperative oral fluoropyrimidine-based chemoradiation (CRT) followed by postoperative adjuvant fluoropyrimidine-based chemotherapy (CT) improves disease-free survival (DFS) in locally advanced rectal cancer. Methods: Between 11/2008 and 09/2011, patients with rectal adenocarcinoma within 12 cm from the anal verge, T3/4 and/or node-positive, with no evidence of metastatic disease and considered either resectable at the time of entry or expected to become resectable, were randomly assigned to receive 5 weeks of preoperative CRT with capecitabine, followed by 6 cycles of adjuvant CT with capecitabine with (arm 2) or without (arm 1) the addition of oxaliplatin before and after surgery. 440 DFS events were required to have 80% power to detect an improvement in 3-year DFS from 65% with capecitabine alone to 72% with capecitabine and oxaliplatin (HR = 0.763) using a two-sided alpha of 5% and owing for an interim analysis for early efficacy at 200 events. The primary analysis was intentto-treat, adjusted for stratification factors (clinical T category, nodal status, distance from the tumor to the anal verge and method of locoregional staging) except center. Results: 1094 patients randomized (547 each arm); 543 eligible patients (arm 1), 526 (arm 2) started treatment; 67.4% completed protocol treatment in arm 1 vs. 53.8% in arm 2. Early analysis driven by the IDMC did not show a difference in 3 years in DFS and OS between both arms.

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The final analysis 01/16 after a median follow-up of 52 months (86–55 months) again are not showing a difference. Subgroup analysis showed superiority for DFS in nongerman participants (33%), whereas for german participants (67%) showed an inferior outcome, cape vs. cape/ox; 78,2% vs. 73,6% (arm1); 73,2% vs. 81,1% (arm 2). Multivariate analysis for identification of the confounding factors is ongoing and will be presented. Conclusion: PETACC-6 does not show any benefit for the addition of oxaliplatin to capecitabine in contrast to the AIO / ARO / CAO trial and the Korean trial. Disclosure: No conflict of interest disclosed. V645

TAS-102 versus placebo plus best supportive care in patients with metastatic colorectal cancer refractory to standard therapies: Final survival results of the phase III RECOURSE trial Karthaus M.1, Van Cutsem E.2, Mayer R.J.3, Ohtsu A.4, on behalf of the RECOURSE Study Group Klinikum Neuperlach, Hämatologie und Onkologie, München, Germany, University Hospital Leuven, Leuven, Belgium, 3Dana Farber Cancer Center, Boston, United States, 4National Cancer Center Hospital East, Chiba, Japan 1 2

Introduction: TAS-102 is comprised of an antineoplastic thymidine-based nucleoside analog, trifluridine, and a thymidine phosphorylase inhibitor, tipiracil hydrochloride. Efficacy and safety of TAS-102 in patients with metastatic colorectal cancer refractory/intolerant to standard therapies were evaluated in the RECOURSE trial. Original results of RECOURSE based on the cut-off date of January 24, 2014, in which 72% of mortality events had occurred, demonstrated a significant improvement in overall survival (OS) with TAS-102 vs placebo. Here we report results ofan updated survival analysis from RECOURSE and a retrospective analysis of OS outcomes based on a clinical prognostic risk index. Methods: Patients were randomized 2:1 to receive TAS-102 (n = 534) or placebo (n = 266). Study treatment continued until disease progression, death, or unacceptable toxicity. The primary endpoint was OS; final survival data were collected on October 8, 2014. A clinical OS prognostic risk score was assessed to evaluate OS effect in patients from low to high prognostic score. Prognostic factors contributing to the risk index were from those prespecified in the multivariate modeling assessment of OS outcome. Results: At data cut-off for the final survival analysis, 89% of the 800 patients randomly assigned to TAS-102 or placebo had died, accounting for 138 events in addition to 574 (72%) events included in the original analysis. Survival at 1 year was 27.1% vs 16.6% for TAS-102 vs Placebo, respectively. Median OS was 7.2 mo in the TAS-102 arm vs 5.2 for pts receiving Placebo HR 0.69 (CI 0.59–0.81) with a p-value < 0.0001. Overall, the toxicity profile for these long-term treated patients did not appear different from that which has already established in the overall population. The updated results for OS are shown in the Table. Tab. 1.

Median OS, mo (95% CI) Hazard ratio (95% CI) P value (1-sided) 1-year survival, % (95% CI)

Original Analysis TAS-102 Placebo (n = 534) (n = 266)

Updated Analysis TAS-102 Placebo (n = 534) (n = 266)

7.1 (6.5–7.8)

5.3 (4.6–6.0)

7.2 (6.6–7.8)

5.2 (4.6–5.9)

0.68 (0.58–0.81)

0.69 (0.59–0.81)

<0.0001

26.6 (22.2–31.1)

17.6 (12.7–23.1)

27.1 (23.3–30.9)

16.6 (12.4–21.4)

Conclusions: An updated survival analysis confirmed that OS benefit with TAS-102 was maintained and increased to a full 2 months; improvement in 1-year survival surpassed 10% in these heavily pretreated patients. OS

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benefit appears to be maintained for all patients in the trial regardless of prognostic status at trial entry. Clinical trial information: NCT01607957 ( Reused with permission from the American Society of Clinical Oncology (ASCO).

Lack of oxaliplatin dose adjustment is correlated with an increased frequency of oxaliplatin-induced neuropathy in obese patients with metastatic colorectal cancer (mCRC)

This abstract was accepted and previously presented at the 2016 ASCO-GI symposium).

Stocker G.1, Giessen-Jung C.2, Schmiegel W.3, Lordick F.1, Hacker U.1, Heinemann V.2

Disclosure: Meinolf Karthaus: Advisory Role: Taiho, Servier; Financing of Scientific Research: Taiho, Servier; A. Ohtsu: Honoraria: Taiho

V646

Cost-effectiveness of systemic therapies for metastatic colorectal cancer according to ESMO: “Magnitude of clinical benefit score” Kaufmann J.1, Strehblow M.1, Wappl M.1, Maier B.1, Schaller G.1 KA-Rudolfstiftung, 4. Med.Abteilung FA Onkologie und Hämatologie, Wien, Austria 1

Introduction: The Magnitude of Clinical Benefit Scale (MCBS) evaluated the magnitude of the benefit of new drugs in solid tumors (Cherny et al, Ann Oncol, 2015). We evaluated the options from the MCBS and compared them to the current ESMO guidelines in metastatic colorectal cancer (mCRC) (Van Cutsem, 2014) for overall survival (OS) and cost. Methods: Costs were calculated based on the Austrian drugs price, body surface area of 1.79m² (Sacco, PLoSOne, 2009) and 75kg of weight. Progression-free survival (PFS) was used to calculate the treatment duration for each line. The OS survival data from Tournigand (Tournigand, 2004) were chosen as baseline. The drugs with a score ≥3 in mCRC in the MCBS were used. Therapeutic strategies with the same magnitude of benefit were applied. If not feasible, the OS from the trial was used and compared to the Tournigand data according to the chemotherapy (CT) backbone. The sequences from the ESMO guidelines were evaluated in term of OS (sum of the different PFS) and compared to the Tournigand data. For the sequences, PFS were used for the treatment duration and the associated treatment costs. ICER and cost/increased month in term of OS were evaluated for each approach. Results: Based on the MCBS, 3 combinations had a score ≥3 in 1st line mCRC: IFL+bevacizumab (B) (Hurwitz, 2004), FOLFIRI + cetuximab (C) (Van Cutsem, 2015) and FOLFOX + panitumumab (P) (Douillard, 2013). In 2nd line and 3rd line, only the FOLFIRI+ P(Peeters, 2015) and C (Karapetis, 2008), respectively, had a score ≥3. Based on the inclusion criteria from these trials, no sequencing strategy with a score ≥3 can be applied. None of the recommended strategy of the ESMO guidelines is scoring within the MCBS, nevertheless the cost/month gained in OS, when applying the ESMO guidelines, are in line with the gain from the MCBS Conclusions: The use of the MCBS with a score ≥ 3 does not allow a mCRC treatment strategy. Some recommended combinations (IFL + B) are less efficient than CT alone with major additional costs. For the ESMO strategies, it seems that costs/month gained for Ras wild type patients are more controlled when starting with anti-EGFR in 1st line. When an anti-angiogenic beyond progression strategy is applied, changing from B to aflibercept seems to reduce the costs/month gained (versus B beyond progression). Full costs data and ICER will be presented. Disclosure: Johann Kaufmann: Financing of Scientific Research: Amgen GmbH, Bayer GmbH, Cellgene GmbH, Eli Lilly GmbH, Gilead GmbH, Roche GmbH, Sanofi-Aventis GmbH, Servier; Other Financial Relationships: Amgen GmbH, Bayer GmbH, Cellgene GmbH, Eli Lilly GmbH, Roche GmbH, Sanofi-Aventis GmbH, Servier Georg Schaller: Financing of Scientific Research: Amgen Gmbh, Roche GmbH; Other Financial Relationships: Amgen Gmbh, Roche GmbH, Boehringer-Ingelheim Gmbh

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University Cancer Center Leipzig (UCCL), University Medicine Leipzig, Leipzig, Germany, 2Department of Medical Oncology, Klinikum Grosshadern and Comprehensive Cancer Center, University of Munich, Munich, Germany, 3 Knappschaftskrankenhaus, Medical Department, Ruhr University Bochum, Bochum, Germany

Introduction: Chemotherapy (Ctx.) doses are usually calculated according to the body surface area (BSA). In clinical practice, dose capping at 2m² BSA represents a widely used approach. ASCO-guidelines recommend to dose Ctx. according to the actual BSA and to avoid dose capping. We investigated if neuropathy in obese patients is more frequent when oxaliplatin was fully dosed. Methods: We performed a retrospective combined analysis of two prospective randomized controlled trials of first-line Ctx. in mCRC: Both trials compared capecitabine plus oxaliplatin (CAPOX) with capecitabine plus irinotecan (CAPIRI) combined with either cetuximab or bevacizumab. In total, 88 patients received CAPOX plus cetuximab and 127 patients received CAPOX plus bevacizumab. Oxaliplatin was given at a dose of 130mg/m² BSA every 3 weeks. We calculated BSA using the Mosteller formula (√height [cm] x weight [kg]/ 3600) and Body mass index (BMI), with body height and weight measured at the first day of Ctx. We categorized patients into 5 BMIgroups: < 20kg/m²; 20.0–21.9kg/m²; 22.0–24.9kg/m²; 25.0–27.9kg/m² and ≥28.0kg/m². The actually given oxaliplatin dose was compared with a theoretical absolute dose based on a calculation according to BSA without dose capping. Patients were categorized as fully dosed if 95–105% of the theoretical oxaliplatin dose was given. Less than 95% was considered as capped/ reduced dose. More than 105% of the theoretical dose was considered overdosed and patients were excluded from further analysis. Results: Of 215 patients who received CAPOX, 25 (12%) were categorized dose reduced, 181 (84%) fully dosed and 3 (1%) overdosed. In 6 (3%) patients the absolute oxaliplatin dose given was not available. 69 (32%) patients developed neuropathy CTC II° or III°. Patients with a BMI < 20kg/ m² showed a lower frequency of neuropathy (15% vs. 30–37%). Of the 40 patients with a BMI ≥ 28kg/m² and fully dosed oxaliplatin, 16 (40%) developed neuropathy. Of the 21 patients with a BMI ≥ 28kg/m² and dose reduced oxaliplatin, 6 (29%) developed neuropathy. Conclusions: Current recommendations to avoid Ctx. dose capping in obese patients is based on the fact that hematological and GI toxicity are not increased but survival outcomes may be worse. Our data suggest that a correlation between higher oxaliplatin doses and neuropathy exists in obese patients. More data is needed regarding Ctx. dosing in overweight and obese patients regarding distinct side effects in this group of patients. Disclosure: Gertraud Stocker: No conflict of interest disclosed. Volker Heinemann: Advisory Role: Merck, Roche,Sanofi, Amgen, SIRTEX, Baxalta, Lilly; Financing of Scientific Research: Merck, Roche, Sanofi, Amgen, SIRTEX, Baxalta, Lilly; Expert Testimony: Merck, Roche, Amgen, SIRTEX

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V648

Interim analysis of the non-interventional QoLiTrap study – Quality of life, therapy management and progression-free survival data with Aflibercept and FOLFIRI from clinical practice in Germany, Austria and Switzerland Scholten F.1, Derigs G.1, Losem C.2, Kröning H.3, Windemuth-Kieselbach C.4, Hoffmann C.M.5, Zaun S.5, Thaler J.6, von Moos R.7, Hofheinz R.-D.8 Klinikum Frankfurt Höchst, Frankfurt, Germany, 2Praxis für Hämatologie und Onkologie, Neuss, Germany, 3Schwerpunktpraxis für Hämatologie und Onkologie, Magdeburg, Germany, 4Alcedis GmbH, Gießen, Germany, 5Sanofi Aventis Deutschland GmbH, Berlin, Germany, 6Klinikum Wels-Grieskirchen, Wels, Austria, 7Universitätsklinikum Graubünden, Chur, Switzerland, 8 Universitätsmedizin Mannheim, Mannheim, Germany 1

Introduction: QoLiTrap is a non-interventional study to assess the quality of life (QoL) and clinical outcomes of patients with metastatic colorectal cancer (mCRC) treated with Aflibercept and FOLFIRI in Germany, Austria and Switzerland. In this interim analysis we report a data set of 402 patients. Methods: Patients included in QoLiTrap were treated with FOLFIRI and Aflibercept according to the label. Quality of life (QoL) was assessed by EORTC-QLQ C30 questionnaires at baseline and before every therapy cycle. Clinical data (overall survival, progression free survival, response to treatment, therapeutic sequences) were reported by participating physicians. Results: Patients eligible for this analysis (n = 402) had filled out at least the baseline and 2 additional QoL-questionnaires. These patients had a median age of 65 years and over 80% had an ECOG PS of 0 or 1. Around half of the patients had acquired a RAS-mutation (49% RAS mut., 40% RAS wt., 11% N/A). Aflibercept was predominantly given in 2nd line treatment (48%) but also in 1st (9%), 3rd (20%) and later therapy lines with a mean cycle number of 8.26 over all therapy lines. In 20% of all patients with response documentation (n = 221) a complete or partial response was achieved, while stable disease was reported in 52% and progressive disease in 28%, resulting in a tumor control rate of 72%. Median progression-free survival was similar in anti-EGFR- (mPFS: 7.11; CI 3.72- 12.47; n = 42) and Bevacizumab-pretreated patients (mPFS: 7.30; CI 5.79- 8.75; n = 169). Side effects were manageable, most frequent grade 3/4 events were hypertension (12.4% of patients) and diarrhea (5% of patients). Global health status declined moderately (mean change in EORTC-score from baseline to week 12: -10 (n = 378). Conclusion: This interim analysis of the non-interventional QoLiTrap study provides deep insights into the clinical practice with Aflibercept in Germany, Austria and Switzerland. Aflibercept and FOLFIRI are embedded in varying therapeutic sequences and achieve a good tumor control rate while conserving patients’ quality of life due to manageable side effects. The data indicate that the effect of treatment is independent of a pre-treatment with either Bevacizumab or anti-EGFR agents. This study is funded by Sanofi. Disclosure: Felicitas Scholten: No conflict of interest disclosed. Ralf-Dieter Hofheinz: Financing of Scientific Research: Sanofi-Aventis, Roche, Merck, Amgen, Bayer, medac; Expert Testimony: Sanofi, Roche, Merck, Amgen, medac

Fortbildung

Keimzelltumoren V651

Refractory germ cell tumors: Molecular mechanisms of resistance and targeted treatment approaches Oing C.1, Bokemeyer C.1 University Medical Center Hamburg-Eppendorf, Department of Oncology, Hematology and Stem Cell Transplantation with Section of Pneumology, Hamburg, Germany 1

Introduction: Germ cell tumors are the most common malignancy among young men aged between 15 to 45 years. Despite overall high cure rates of >90%, 5% to 10% of all patients develop platinum-resistance and die of their cancer. Cisplatin-resistance appears to be of multifactorial origin, rather than a consequence of distinct aberrations. Methods: PubMed, Medline, and ASCO and ESMO conference proceedings were comprehensively searched to identify articles on resistance mechanisms, targetable structures and targeted treatment approaches in cisplatin-resistant germ cell tumors. Results: Patients suffering platinum-refractory disease and/or multiple relapses have a dismal prognosis with an overall survival of usually a few months, only. Cisplatin-sensitivity has been linked to the expression of wild-type p53, whereas TP53 mutations or amplification of the wtp53 suppressor MDM2 occur in 25% of resistant tumors. Moreover, the BRAF V600E mutation was exclusively found in 26% of resistant tumors, which highly correlated with mismatch repair deficiency. Another elicitor of cisplatin-resistance may be over-activation of the PIK3CA-AKT signaling pathway, either by PIK3CA or AKT mutations or expression loss of their negative regulator PTEN. To date, molecularly targeted treatments failed to display clinically meaningful efficacy in small clinical trials, since most of the trials did not select patients by drugable biomarkers. Individual patients achieved objective responses undergoing treatment with the two most active agents so far: sunitinib (13%), or the CD30-directed antibody-drug conjugate brentuximab vedotin (22%), but responses were short-lived. Notably, systemic targeting of CDK 4/6 by palbociclib yielded disease stabilizations in a relevant proportion of patients with retinoblastoma protein-expressing teratoma and teratoma with malignant transformation. Conclusion: There is a need to further improve long-term outcomes by new systemic treatment approaches in the refractory setting. However, clinical trials evaluating targeting agents in refractory germ cell tumors have been disappointing, so far. The design of larger clinical trials is difficult due to the high curability by standard chemotherapy and the heterogeneity of the disease. Identification of drugable biomarkers and marker-stratified treatment approaches may aid to improve outcomes. Particularly, the potential of novel immune therapeutic agents, i.e. immune-checkpoint inhibitors, remains open yet. Disclosure: No conflict of interest disclosed.

Expertenseminar

Naturheilverfahren – Integrative Onkologie V654

From naturopathy to integrative oncology Hübner J.1 Deutsche Krebsgesellschaft, Berlin, Germany

Methods of complementary (accompanying and supplementary methods) and alternative (instead of conventional methods) medicine are widely used in oncology. Complementary medicine often is accepted without critical appraisal as it allegedly is in agreement with patients´ desire for a “soft” treatment, may

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support mentally and will for sure do no harm. Most often, oncologists leave the field to other “specialists” for naturopathy, without there being any consent on quality of the counseling or treatment. In contrast, with evidence based knowledge on the most often used methods of complementary and alternative medicine, oncologists may integrate counseling in regular communication without much more need of time. This would lead to more safety in using complementary methods as supportive treatment. Side effects as well as interactions in fat are the main risks of complementary medicine. The decisive problem is, that evidence of the benefits and risks most often is scarce. Therefore, physicians and nurses should be attentive to all effects and symptoms of their patients. Risks of using complementary and more so alternative methods are costs, time spent for the treatment and even raising false hope. If we conceive complementary medicine as the answer to the question of patients and caregivers, what they may contribute to improving the situation, complementary medicine has large potential. Valuable complementary methods may be applied by the patient himself/herself, on his own decision and in his/her own responsibility, thus allowing for autonomy. Disclosure: Jutta Hübner: Employment or Leadership Position: Deutsche Krebsgesellschaft; Advisory Role: Verschiedene Sozialgerichte; Expert Testimony: Fa. Loges; Immaterial Conflict of Interests: Vorsitzende der Arbeitsgemeinschaft Prävention und Integrative Onkologie

Freier Vortrag Akute Leukämien V655

Mutant p53 as a novel therapeutic target in Acute Lymphoblastic Leukemia Demir S.1,2, Selivanova G.3, Tausch E.4, Wiesmüller L.5, Stilgenbauer S.4, te Kronnie G.6, Debatin K.-M.1, Meyer L.H.1 Universitätsklinikum Ulm, Klinik für Kinder- und Jugendmedizin, Ulm, Germany, 2Internationale Graduiertenschule für Molekulare Medizin, Universität Ulm, Ulm, Germany, 3Karolinska Institute, Department of Microbiology, Tumor and Cell Biology, Stockholm, Sweden, 4Universitätsklinikum Ulm, Klinik für Innere Medizin III, Ulm, Germany, 5Universitätsklinukum Ulm, Klinik für Frauenheilkunde und Geburtshilfe, Ulm, Germany, 6University of Padova, Department of Women`s and Children`s Health, Padova, Italy 1

Introduction: Mutations of the tumor suppressor gene TP53 (TP53mut) are present in acute lymphoblastic leukemia (ALL) at lower frequencies but associated with therapy resistance and relapse. The small molecule APR-246, which targets mutated p53, has shown activity in different TP53mut malignancies. However, targeting mutant TP53 has not been addressed in ALL so far. Methods: A large cohort of patient-derived xenograft samples established in our NOD/SCID/huALL mouse model from pediatric B cell precursor (BCP)-ALL cases was analyzed for TP53mut (chromatography and Sanger sequencing). Activity of APR-246 (kindly provided by Aprea, Stockholm, Sweden) was evaluated by half maximal inhibitory concentrations (IC50) and combination indices (CI). Apoptosis was assessed by caspase-3 activation and Annexin-V/propidium iodide positivity, p53 activation by expression of p53 downstream molecules by western blot analysis. p53 deficient cell lines were generated by lentiviral shRNA mediated knock-down. Results: Of 62 BCP-ALL samples, 4 were TP53mut (6.4%), corresponding to frequencies reported in patient cohorts. Of 6 BCP-ALL cell lines investigated, 2 were TP53mut. Exposure to the DNA damaging agent doxorubicin showed insensitivity for TP53mut but clear cell death in wild type (TP53wt) leukemias (cell lines and primografts), corroborating p53’s tumor-suppressive function in ALL. Most interestingly, TP53mut but not TP53wt ALL showed a high sensitivity for APR-246. Induction of caspase-3 and positivity for Annexin-V/propidium-iodide revealed apoptosis as mechanism of APR-246 mediated cell death. Importantly, silencing of TP53 abrogated APR-246 induced cell death, indicating that

Abstracts

the activity of APR-246 is dependent on p53. Moreover, induction of p53’s transcriptional targets NOXA and PUMA was observed in response to APR-246 in TP53mut but not TP53wt ALL, indicating functional restoration and activation of p53. Based on these findings, we investigated the ability of APR-246 to re-sensitize TP53mut, DNA-damage resistant ALL. Combinations of APR-246 and doxorubicin led to strong synergism and re-sensitization for DNA-damage induced cell death. Conclusions: Taken together, APR-246 specifically targets TP53mut BCPALL leading to re-activation of p53, NOXA and PUMA mediated apoptosis, and re-sensitization of DNA-damage resistant TP53mut ALL. Thus, targeting and re-activation of mutated p53 provides a promising novel therapeutic strategy in this high-risk subtype of BCP-ALL. Disclosure: No conflict of interest disclosed. V656

Intrinsic and synergistic activity of ABT-199 in B-cell precursor acute lymphoblastic leukemia Seyfried F.1, Demir S.1, Hörl R.1, Zinngrebe J.1, Scheffold A.2, Köhrer S.1, Stilgenbauer S.2, Debatin K.-M.1, Meyer L.H.1 Ulm University Medical Center, Department of Pediatrics and Adolescent Mendcine, Ulm, Germany, 2Ulm University Medical Center, Department of Internal Medicine III, Ulm, Germany 1

Introduction: In acute lymphoblatic leukemia (ALL), deficient cell death pathways are associated with therapy resistance and treatment failure. Anti-apoptotic Bcl-2 family proteins regulate apoptosis and therefore serve as promising targets for novel, directed therapies. ABT-199 binds to Bcl-2, releases pro-death Bcl-2 family molecules, leads to apoptosis induction, and demonstrated anti-leukemia activity. Here, we investigated the activity of ABT-199 on B cell precursor (BCP)ALL including pre-clinical in vivo evaluation and investigated synergistic effects upon combination with conventional chemotherapy. Methods: The effects of different drugs or combinations on BCP-ALL (n = 6 cell lines, n = 16 patient-derived xenografts, pdx) were investigated by half maximal inhibitory concentrations (IC50) and combination indices (CI). Anti-leukemia activity of ABT-199 was analyzed in vivo. Expression of apoptosis regulators was detected by western blot analysis. Mcl-1 deficient cell lines were generated by CRISPR/Cas gene editing. Results: The majority of BCP-ALL samples (4 of 6 BCP-ALL cell lines and 14 of 16 pdx samples) showed high sensitivity to ABT-199 with nanomolar IC50 values. Of note, PBMCs from healthy donors were insensitive to ABT-199. High levels of Bcl-2 and low Mcl-1 expression was detected in ABT-199 sensitive leukemias, whereas ABT-199 resistant leukemias showed high Mcl-1 but low Bcl-2 expression. Interestingly, CRISPR/Cas9-mediated knockout of Mcl-1 clearly sensitized ABT-199 insensitive cell lines for ABT-199 mediated cell death, showing Mcl-1’s role as mediator and marker for ABT-199 resistance. Upon co-exposure with ABT-199 and chemotherapeutic agents used for remission induction (vincristine, asparaginas and dexamethasone) a clear synergistic effect was observed, both in cell lines and pdx ALL. Moreover, in a preclinical setting, recipient animals bearing a high-risk, t(11;19) leukemia were treated with ABT-199 leading to a clear reduction of leukemia load upon ABT-199 in vivo therapy compared to vehicle treated recipients (p < 0.01). Conclusion: Taken together, our data show apoptosis induction and high efficacy of ABT-199 as single compound, both ex vivo and pre-clinically in vivo. Mcl-1 was identified as mediator and indicator for ABT-199 resistance. Importantly, we observed synergy upon combination with conventional induction chemotherapy. Thus, ABT-199 is a promising addendum sensitizing leukemia cells for conventional chemotherapy. Disclosure: No conflict of interest disclosed.

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V657

V658

Frequency and prognosis of BCR-ABL1-like ALL among adult ALL cases

Inotuzumab ozogamicin (InO) for relapsed/refractory (r/r) acute lymphoblastic leukemia (ALL) in the global phase 3 randomized controlled ino-vate trial: efficacy and safety by age and prior therapy

Fasan A.1, Kern W.1, Haferlach T.1, Haferlach C.1 MLL Münchner Leukämielabor, München, Germany

Introduction: In the 2016 revision to the World Health Organization (WHO) classification of myeloid neoplasms and acute leukemia (Arber et al., Blood 2016), Philadelphia chromosome-like acute lymphoblastic leukemia (BCR-ABL1-like ALL) has been included as new provisional entity within B-cell ALL (B-ALL). It is associated with activating lesions of either the ABL1 or JAK signaling pathways and an adverse prognosis. Diagnosis of BCR-ABL1-like ALL is challenging and frequency and outcome in older adults is largely unknown. Methods: The study cohort consisted of 215 adult precursor B-ALL patients (pts) with 132 of these cases lacking the recurrent aberrations BCRABL1, MLL-AF4 and E2A-PBX1 (age 18–85 years, median 54 years). Precursor B-ALL was diagnosed by immunophenotyping in all cases. Gene expression profiles were generated using Affymetrix HG-U133 plus 2.0. BCR-ABL1-like cases were identified according to Roberts et al, NEJM 2014. Results: Overall, 33/215 (15%) B-ALL cases were identified as BCRABL1-like ALL according to gene-expression profiles. Incidence of BCRABL1-like ALL according to age was detected as follows: 1% adolescents (age: 16–20 years); 4% young adults (age: 21–39 years), 2% adults (age: 40–55 years) and 8% older adults (age: 56–90 years), indicating an incidence of BCR-ABL1-like ALL of 10% in adults aged ≥40 years. Regarding genetic alterations, a total of 25/33 cases with BCR-ABL1-like ALL had CRLF2 overexpression (76%). 16/25 cases (64%) with elevated CRLF2 expression harbored IGH-CRLF2 rearrangement. Additionally, 4 pts with rearrangements involving TKI sensitive genes ABL1 (SNX2-ABL1; n = 1), and PDGFRB (EBF1-PDGFRB; n = 3) were identified. Mutations in JAK2 were detected in 16/33 (48%) BCR-ABL1-like ALL pts, all of which were characterized by CRLF2 rearrangements and/or overexpression of CRLF2. NRAS mutations were found in three pts, one pt harbored a KRAS mutation. Furthermore, we found a high frequency of IKZF1 deletions in BCRABL1-like ALL pts (n = 22/33; 67%). Regarding outcome, BCR-ABL1-like ALL pts as compared to BCR-ABL1 positive ALL pts had significantly shorter event-free survival (median 11 vs 28 months; p = 0.040) and overall survival (median 24 months vs n.r.; p = 0.013) Conclusions: 1) BCR-ABL1-like ALL can be detected in adult B- ALL at a frequency of ~10%; 2) BCR-ABL1-like ALL is characterized by a high frequency of kinase-activating lesions; 3) Adult BCR-ABL1-like ALL is correlated with poor prognosis. Disclosure: No conflict of interest disclosed.

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Stelljes M.1, Kreuzer K.-A.2, Deangelo D.J.3, Advani A.S.4, Gökbuget N.5, Liedtke M.6, Stock W.7, Martinelli G.8, O’Brien S.9, Wang K.10, Wang T.11, Paccagnella M.L.11, Sleight B.11, Vandendries E.12, Jabbour E.J.13, Kantarjian H.M.13 Universitätsklinikum Münster, Münster, Germany, 2University of Cologne, Cologne, Germany, 3Dana-Farber Cancer Institute, Boston, United States, 4 Cleveland Clinic, Cleveland, United States, 5Goethe University, Frankfurt, Germany, 6Stanford Cancer Institute, Stanford, United States, 7University of Chicago, Chicago, United States, 8Institute Seràgnoli, DIMES, University of Bologna, Bologna, Italy, 9University of California, Irvine, Orange, United States, 10 Pfizer Inc, Pearl River, United States, 11Pfizer Inc, Groton, United States, 12Pfizer Inc, Cambridge, United States, 13MD Anderson Cancer Center, Houston, United States 1

Introduction: InO, an anti-CD22 antibody-calicheamicin conjugate, showed superior response vs standard care for R/R ALL in the phase 3 INO-VATE trial (complete remission [CR]/CR with incomplete hematologic recovery [CRi], 81% [95% CI; 72–88]; minimal residual disease [MRD] negativity in responders, 78% [68–87]; median remission duration [DoR], 4.6 [3.9–5.4] mo). Here, we assess the efficacy and safety of InO in patients (pts) aged ≥55 vs < 55 y and by prior therapy (salvage [S] 1 vs S2; with vs without (w/o) prior stem-cell transplant [SCT]). Methods: Per protocol, analyses of CR/CRi included the first 218 of 326 pts randomized (ITT218). The safety population included 139 pts who received ≥1 InO dose (max 1.8 mg/m2/cycle [0.8 mg/m2 on d 1; 0.5 mg/m2 on d 8 and 15 of a 21–28 d cycle for ≤6 cycles]). Results: As of October 2, 2014 (trial ongoing), 109 pts in the ITT218 received InO (median age, 47 [range, 18–78] y; pts ≥55 y, n = 43 [39%]; 67% and 32% of pts received InO as S1 and S2 [missing, n = 1]). No statistically significant differences were found in CR/CRi and MRD negativity rates, or DoR between age groups and S1 vs S2 (Table). CR/CRi rates in pts with (n = 17) and w/o (n = 92) prior SCT were 77% (95% CI, 50–93) and 82% (72–89). In the safety population, gr ≥3 adverse events (AEs) were most frequently cytopenias which were more common in pts ≥55 vs < 55 y. Any grade hepatobiliary AE rates were similar in pts ≥55 vs < 55 y (both 26%), significantly higher in S2 vs S1 (47 vs 17%; P < 0.001) and higher in pts with vs w/o prior SCT (42 vs 23%). Veno-occlusive liver disease (VOD), including post-SCT VOD, occurred in 11% each of pts ≥55 vs < 55 y, 8% of S1 vs 16% of S2 pts and 21% with vs 9% w/o prior SCT. More mature DoR data as well as survival data for the overall population and by randomization stratification factors (age < 55 vs ≥55 y, duration of first CR < 12 vs ≥12 months, S1 vs S2) will also be presented. Conclusion: InO was effective in older pts with R/R ALL and as both S1 and S2 therapy. Safety profiles were similar in older vs younger pts; however, hepatotoxicity risk increases with number of prior therapies and prior SCT.

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Tab. 1.

Conclusion: These preclinical data indicate that concomitant administration of antibody therapy and chemotherapy or the administration of the antibody in an MRD-situation may represent promising approaches and suggest a high therapeutic potential of Fc-engineered CD19 antibodies in the therapy of ALL. Disclosure: No conflict of interest disclosed. V660

miR-143 targets ERK5 in granulopoiesis and predicts outcome of Acute Myeloid Leukemia patients Hartmann J.-U.1, Bräuer-Hartmann D.1, Kardosova M.2, Wurm A.A.1, Gerloff D.1, Katzerke C.1, Bill M.1, Schwind S.1, Niederwieser D.1, AlberichJorda M.2, Behre G.1 University Leipzig, Division of Hematology and Medical Oncology, Leipzig, Germany, 2Institute of Molecular Genetics AS CR, v.v.i., Laboratory of Haematooncology, Prague, Czech Republic 1

Disclosure: Matthias Stelljes: Advisory Role: Pfizer; Financing of Scientific Research: Pfizer; Expert Testimony: Pfizer; Hagop Kantarjian: Expert Testimony: Pfizer, Amgen, Astex, Bristol-Myers Squibb, Novartis V659

An Fc-engineered CD19 antibody engages NK cells and macrophages and is effective in Xenograft models of acute lymphoblastic leukemia Kellner C.1, Alsadeq A.2, Cario G.2, Vieth S.2, Roßkopf S.1, Burmeister Y.1, Klausz K.1, Valerius T.1, Schrappe M.2, Gramatzki M.1, Peipp M.1, Schewe D.M.2 Christian Albrechts University of Kiel, 2nd Department of Medicine, Division of Stem Cell Transplantation and Immunotherapy, Kiel, Germany, 2Christian Albrechts University of Kiel, Pediatric Hematology/Oncology, ALL-BFM Study Group, Kiel, Germany 1

Introduction: CD19 represents a promising target antigen for therapeutic antibodies in the treatment of B-lineage acute lymphoblastic leukemia (ALL). However, conventional CD19 antibodies exert limited effector functions. These shortcomings may be overcome by next generation CD19 antibodies with an enhanced potency to trigger effector cell functions as antibody-dependent cell-mediated cytotoxicity (ADCC) or phagocytosis. Here, we tested an Fc engineered CD19 antibody for its ability to engage effector cells and analyzed it in preclinical minimal residual disease (MRD) models of pediatric ALL. Methods: An Fc engineered CD19 antibody and its native IgG1 variant were generated according to published sequences. ADCC was analyzed in 51Cr release assays with natural killer (NK) cells. Phagocytosis was determined by fluorescence microscopy analysis. Patient-derived xenografts of 2 infants with highly aggressive MLL-rearranged ALL were established by intrafemoral transplantation in NOD-SCID-gamma-/- (NSG) mice. The CD19 antibody was applied by consecutive intraperitoneal injections (1mg/kg body weight). Antibody therapy was initiated alone or in combination with a regimen mimicking standard ALL induction chemotherapy (Dexamethasone, Vincristine, L-Asparaginase). Results: The Fc engineered CD19 antibody induced NK-cell mediated lysis of patient-derived ALL cells and efficiently triggered phagocytosis by macrophages. The native CD19 IgG1 antibody was not efficient. Importantly, the Fc engineered CD19 antibody prolonged survival of ALL xenograft mice when low cell numbers were transplanted and therapy was immediately initiated. Analysis of bone marrow from treated surviving mice for patient-specific Ig-rearrangements and the respective MLL-fusion revealed that 3/5 mice in each patient were MRD-negative. When antibody therapy was initiated after leukemia development, the Fc engineered antibody was still efficient, but the effects were less pronounced. Remarkably, the combination of the antibody and cytoreductive chemotherapy resulted in dramatically improved survival rates of 90% as compared to mice treated with the antibody or chemotherapy alone.

Abstracts

Introduction: MicroRNAs, a class of small non-coding RNAs, have been reported to play an important role in the regulation of cellular processes. Deregulation of certain microRNAs may lead to disrupted cellular pathways and thereby to the promotion of leukemic phenotypes. Methods: In this study, we used primary cells, AML-patient samples and AML cell lines to analyze microRNA and protein levels. In vitro studies were performed by transient and stable overexpression or knock down of miR-143. Analysis was performed by PCR, western blot, proliferation assay and measurement of granulocytic surface markers. AGO-RIP was performed to identify and prove microRNA binding to 3`UTRs of target mRNAs. Results: By treatment of CD34+ progenitor cells with G-CSF, we identified miR-143 among others as strongly upregulated during granulocytic differentiation. We could also show the upregulation of miR-143 in APL patients upon ATRA treatment as well as in AML cell lines undergoing granulocytic differentiation. Analysis of the TCGA database of 188 AML patient samples showed that high miR-143 expression is associated with better overall survival. In addition, QPCR analysis of pri-miR-143 expression in reduced intensity conditioning (RIC) allogenic transplanted AML patients revealed that high expression of miR-143 is associated with better event-free survival in this group of patients. Functional studies showed that overexpression of miR-143 in K562-C/EBPα-p42 cells enhances C/ EBPalpha induced granulocytic differentiation, significantly. In contrast, stable knock down of miR-143 reduced the ability of K562-C/EBPα-p42 cells to undergo granulocytic differentiation upon β-estradiol treatment. Interestingly, stable knock down of miR-143 in mice showed significantly reduced number of mature granulocytes in peripheral blood. By in silico prediction, we found that miR-143 targets MAPK protein family members (e.g. MAPK7). Western blot analysis of AML patient samples and G-CSF stimulated CD34+ cells clearly showed an inverse correlation of miR-143 and MAPK7 (ERK5) protein expression. Immunoprecipitation of miR143 mRNA targets showed a strong enrichment of ERK5-mRNA upon overexpression of miR-143. Conclusion: Our results highlight miR-143 as an important factor in granulocytic differentiation, whose deregulation may play an important role in the development of AML. Finally, our data propose miR-143 as a good prognostic marker in Acute Myeloid Leukemia. Disclosure: No conflict of interest disclosed.

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Freier Vortrag

Multiples Myelom klinisch V661

Applicability of AL amyloidosis cardiac staging systems in patients with impaired renal function Dittrich T.1, Hegenbart U.1, Kimmich C.1, Kristen A.2, Ho A.D.1, Schönland S.O.1 Universitätsklinikum Heidelberg, Medizinische Klinik V, Heidelberg, Germany, Universitätsklinikum Heidelberg, Medizinische Klinik III, Heidelberg, Germany

1 2

Introduction: Systemic light chain amyloidosis (AL) is a rare and life-threatening disease characterized by systemic deposition of misfolded free light chains. Current staging systems of AL (MAYO2004: Dispenzieri et al. 2004, JCO and MAYO2012: Kumar et al. 2012, JCO) facilitate cardiac biomarkers (troponins, NT-proBNP) and serum free light chains. However, those biomarkers are known to depend on renal function, which is frequently impaired in AL patients due to heart and kidney involvement. Our hypothesis was that the aforementioned widely used staging systems are inappropriate in patients with advanced renal insufficiency. Methods: We retrospectively analyzed 610 patients with pathologically confirmed AL that were treated and followed at the Heidelberg Amyloidosis Center between July 2002 and March 2015. Results: Median overall survival significantly differed in the staging groups of the MAYO2004 (111 vs. 72 vs. 10, months, all p < 0.001) and MAYO2012 (130 vs. 95 vs. 20 vs. 6, months, highest p =0.003) classifications. As expected, reduced estimated glomerular filtration rate (eGFR) was highly associated with increased cardiac troponins (TNI, cTNT, hsTNT) and NT-proBNP (data not shown), but had no effect on serum dFLC levels. Using a threshold of 50 ml/min, stages I and II of the MAYO2004 and MAYO2012 systems were significantly less frequent in patients with reduced eGFR (40% vs. 62% and 30% vs. 50%, both p < 0.001). Moreover, reduced renal function resulted in poor discrimination of stages I and II in both MAYO2004 (p =0.309) and MAYO2012 (p =0.606), while the upper two stages still predicted different overall survival in both MAYO2004 and MAYO2012 (both p < 0.001). Of note, reduced eGFR < 50 ml/min itself was associated with inferior median overall survival (22 vs. 53 months, p < 0.001), while patients with proteinuria >5 g/ day as a hallmark of kidney organ involvement showed longer median overall survival (68 vs. 24 months, p < 0.001). Conclusion: Reduced eGFR results in higher MAYO stages of the respective patients due to accumulation of cardiac biomarkers. However, those higher stages are predictive for inferior overall survival, as impaired renal function itself is associated with inferior overall survival. Notably, proteinuria has no adverse impact on overall survival. As a next step, validation of our data in a different cohort of AL patients is warranted to decide if eGFR should be incorporated in a new staging system of AL amyloidosis. Disclosure: No conflict of interest disclosed. V662

Results of European trial of free light chain removal by extended haemodialysis in cast nephropathy (EULITE) Weisel K.C.1, Cook M.2, Hutchison C.3, Fifer L.4, Bradwell J.5, Heyne N.6, Cockwell P.4 University of Tübingen, Medical Center, Hematology, Oncology and Immunology, Tübingen, Germany, 2Queen Elizabeth Hospital Birmingham, Department of Haematology, Birmingham, United Kingdom, 3Hawke’s Bay District Health Board, Department of Nephrology, Hawke’s Bay, New Zealand, 4 Queen Elizabeth Hospital Birmingham, Department of Renal Medicine, Birmingham, United Kingdom, 5University of Birmingham, College of Medical and Dental Sciences, Birmingham, United Kingdom, 6University of Tübingen, Section of Nephrology and Hypertension, Tübingen, Germany 1

Background: Severe acute kidney injury (AKI) secondary to myeloma cast nephropathy (MCN) and requiring acute dialysis is caused by the

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pathogenic (involved) immunoglobulin serum free light chain (sFLC). High cut-off haemodialysis (HCO-HD) can remove large quantities of sFLC and retrospective uncontrolled clinical trials using HCO-HD have better renal recovery rates than those historically reported without HCOHD. However, it is not known if these better outcomes were associated with HCO-HD or novel chemotherapy regimens. Methods: We carried out a prospective multi-centre RCT in patients with newly diagnosed MM and associated MCN who required acute dialysis treatment, comparing HCO-HD to standard high flux (HF)-HD, and using bortezomib based chemotherapy as the standard of care. Ninety (90) patients with a first diagnosis of MM, MCN confirmed by renal biopsy, sFLC levels of the involved LC ≥500 mg/l (Freelite™), and a requirement for acute dialysis, were randomised to receive HCO-HD or HF-HD. Primary outcome was independence of dialysis at 3-months. Patients were followed for 2-years. Results are reported as intention to treat (ITT). Results: There was no difference at randomisation between groups for age, ethnicity, gender, myeloma type (light chain only vs intact Ig), and sFLC by isotype. There was no difference at 3 weeks for sFLC levels between the two groups. 24/43 patients (55.8%) in the HCO-HD group and 24/47 patients (51.6%) in the HF-HD group recovered renal function by 3 months (not significant, NS). Overall recovery of renal function was 62%; 58.1% in the HCO-HD group, 66% in the HF-HD group (NS). 35% in the HCO-HD group and 32.2% in the HF-HD group received a stem-cell transplant (NS). Overall survival (patients alive and under follow-up) at 2-years was 24 (55.8%) in the HCO-HD group and 36 (76.6%) in the HFHD group. There were more lung infections in the first 3-months in the HCO-HD group (12 vs 3, P = 0.014). Conclusion: In this RCT, where all patients received bortezomib based chemotherapy, 62.2% of patients with MCN recovered renal function and overall survival at 2-years was 66.7% (ITT). HCO-HD, compared to HFHD, did not improve renal recovery rates in patients requiring acute dialysis for severe AKI secondary to MCN. Further data will be presented. Disclosure: Katja Weisel: Advisory Role: Janssen; Financing of Scientific Research: Janssen; Expert Testimony: Janssen Paul Cockwell: No conflict of interest disclosed. V663

Update on an outpatient autologous stem cell transplantation program for patients with multiple myeloma Lisenko K.1, Sauer S.1, Egerer G.1, Goldschmidt H.1,2, Hillengass J.1, Schmier J.-W.1, Witzens-Harig M.1, Ho A.D.1, Wuchter P.1 Universitätsklinikum Heidelberg, Abteilung Innere Medizin V, Heidelberg, Germany, 2Nationales Centrum für Tumorerkrankungen, Heidelberg, Germany 1

Introduction: High-dose (HD) chemotherapy with melphalan and autologous blood stem cell transplantation (ABSCT) is an established treatment option for eligible patients aged < 70 years with advanced stage multiple myeloma (MM). While patients in the USA and Canada undergo this procedure mostly on an outpatient basis, in Germany and Western Europe an inpatient setting has been the standard. We present a follow-up analysis on an outpatient transplant program established in 2012 at our center. Methods: Major inclusion and exclusion criteria for eligibility were defined as follows: patients had to be able to reach the hospital within 45 minutes, had reliable support from their family at home, had an ECOG performance score of 0–1 and were willing and able to comply with the demands of the program. Patients with amyloidosis or severe co-morbidities were not included. All patients were treated and examined on daily visits by a team of physicians. Results: From September 2012 until April 2016, 21 patients diagnosed with MM stage IIIA underwent HD chemotherapy and ABSCT on an outpatient basis. All patients engrafted within the expected time range (neutrophil engraftment 12–20 days, median 14 days) and no severe adverse events occurred. 14 patients (67%) had an episode of fever ≥38.5°C, but only in four patients (19%) blood cultures were found to be positive. Based on clinical findings, either an intravenous antibiotic therapy with ertapenem was initiated and treatment in the outpatient clinic was con-

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tinued or the patient was admitted to the hospital for inpatient i.v. antimicrobial treatment. Although rather liberal criteria for hospital admission were applied, 14 of 21 patients (67%) were treated entirely on an outpatient basis. Seven patients (33%) were temporarily admitted for inpatient treatment with a median duration of five days (range 2–18 days), mainly because of infectious complications. Feedback from patients was obtained by means of a questionnaire and revealed a high level of satisfaction with the outpatient setting. Conclusions: HD chemotherapy and ABSCT on an outpatient basis is safe and feasible if conducted in a comprehensive surveillance program. Feedback from patients was very positive, which led to further continuation and expansion of the program. Disclosure: Katharina Lisenko: No conflict of interest disclosed. Patrick Wuchter: Advisory Role: Teilnahme an Advisory Boards von Sanofi-Aventis.; Financing of Scientific Research: Honorare von Sanofi-Aventis. Travel Grants von Hexal AG. V664

Chemotherapy before and after heart transplantation for patients with very advanced cardiac AL amyloidosis: single centre results with long-term follow-up Haack K.1, Schönland S.2, Kristen A.3, Blum P.3, Kimmich C.2, Katus H.3, Ho A.4, Ruhparwar A.5, Raake P.3, Hegenbart U.2 Universitaetsklinikum Heidelberg, Heidelberg, Germany, 2Universitaetsklinikum Heidelberg, Amyloidose-Zentrum, Heidelberg, Germany, 3Universitaetsklinikum Heidelberg, Kardiologie, Heidelberg, Germany, 4Universitaetsklinikum Heidelberg, Hämatologie/Onkologie/Rheumatologie, Heidelberg, Germany, 5 Universitaetsklinikum Heidelberg, Herzchirurgie, Heidelberg, Germany 1

Introduction: Survival rates for patients with light-chain (AL) amyloidosis are gravely reduced by advanced cardiac involvement with a median survival of 3,5 months in patients with very advanced cardiac involvement. High-dose Melphalan (HDM) and autologous stem cell transplantation (ASCT) cannot be applied to those patients because of a very high risk of therapy-related morbidity and mortality. Heart transplantation (HTx) is a treatment option for those patients. Our aim was to examine the cases of cardiac AL patients who received HTx, combined with chemotherapy and/or HDM/ASCT in our institution. Patients and methods: Data from 38 patients (19 males, 19 females) with very advanced cardiac AL who were treated between 2002 and 2015 were retrospectively analyzed. The patients were high-urgency listed for orthotopic HTx. All data are derived as medians with range or absolute numbers. Results: Median age at diagnosis was 50 years (35–62). Median differential free light chain (dFLC) was 564 (69 - 2752). Median NT-proBNP at diagnosis was 6780 ng/l (1500 - 53194). Patients stayed on the high-urgency waiting list for a median of 26 days (range 3–54) before 2009, and a median of 63 days (15–259) after 2009. Thirty-three patients were treated with chemotherapy prior to HTx (mostly dexa w/o Bortezomib) to reduce dFLC during the waiting time. Within a median time of 5 months after HTx (4–29), 18 patients received ASCT. HDM was used with either 200 mg/m2 (N = 10) or reduced dosage (N = 8) in patients with reduced kidney function. Complete remission (CR) was achieved in 7 patients, (24% of all 29 transplanted pts; very good partial remission (VGPR) in 6 patients (21%) and partial remission (PR) in 7 patients (24%). Twenty-four 24 patients died. Cause of death was either progression of AL (N = 15), sepsis (n = 4), heart transplant rejection (n = 3) or other (n = 2). Seven patients died on the waiting list with a median survival (start point: HU listing) of 16 days. Patients who underwent HTx had a median survival of 46 months (3–175, 1-year survival: 77%). Discussion and conclusion: HTx in combination with chemotherapy is a feasible treatment approach in patients with very advanced cardiac amyloidosis who cannot be treated with a standard chemotherapy. Patients who reach HTx have later on a nearly 50% chance for a very good hematologic remission (VGPR or better) and consecutively a favorable survival probability with a median OS of nearly 4 years in our series.

Abstracts

Disclosure: Katharina Haack: No conflict of interest disclosed. Ute Hegenbart: Financing of Scientific Research: Janssen, Celgene, Binding Site V665

Revlimid, Bendamustine and Prednisolone (RBP) in relapsed/ refractory multiple myeloma: final results of a phase II clinical trial OSHO - #077 Pönisch W.1, Beck J.2, Heyn S.1, Schliwa T.1, Lange T.1, Andrea M.1, Jentzsch M.1, Schwind S.1, Bill M.1, Mohren M.3, Hoffmann F.-A.4, Kreibich U.5, Gläser D.6, Uhlig J.7, Mügge L.-O.8, Kragl B.9, Zehrfeld T.10, Schwarzer A.11, Winkelmann C.12, Edelmann T.13, Niederwieser D.W.1, OSHO – Ostdeutsche Studiengruppe Hämatologie und Onkologie Universitätsklinikum Leipzig, Hämatologie und internistische Onkologie, Leipzig, Germany, 2Martha-Maria Krankenhaus, Hämatologie und Onkologie, Halle / Saale, Germany, 3Johanniter Krankenhaus Stendal, Hämatologie und internistische Onkologie, Stendal, Germany, 4Hematology Practice Leipzig, Leipzig, Germany, 5Heinrich-Braun Krankenhaus Zwickau, Zwickau, Germany, 6 Klinikum Süd Rostock, Hämatologie, Rostock, Germany, 7Hematology Practice Naunhof, Naunhof, Germany, 8Universitätsklinikum Jena, Hämatologie und internistische Onkologie, Jena, Germany, 9Universitätsklinikum Rostock, Rostock, Germany, 10Kreiskrankenhaus Torgau, Hämatologie und internistische Onkologie, Leipzig, Germany, 11Hematology Practice Leipzig, Leipzig-Dösen, Germany, 12Paul Gerhard Stift Wittenberg, Wittenberg, Germany, 13Hematology Practice Schkeuditz, Schkeuditz, Germany 1

Purpose: While the role of lenalidomide monotherapy in the treatment of relapsed/refractory patients with multiple myeloma (MM) is well established, combination therapies with lenalidomide are still under investigation in many phase 2/3 studies. In the current study, a combination therapy of lenalinomide (Revlimid®), bendamustine and prednisolone (RBP) was tested in patients with relapsed or refractory MM. Methods: In the previously completed phase 1 study RPB with a dose of 75 mg/m² bendamustine d 1–2, prednsolone 100 mg d 1–4 and 25 mg lenalidomide d 1–21 was well tolerated in patients with relapsed/refractory MM. Cycles were repeated every 28 days for a maximum of eight cycles. Thereafter, patients received maintenance therapy with 10 mg lenalidomide over 10 cycles. Results: Between July 2011 and September 2013, 25 patients (19 patients of the phase 2 study and additional 6 patients from the highest dose level of the phase 1 study) were included in this analysis. Sixteen patients (64%) had previously undergone autologous SCT, 7 of these patients had previously received a second transplant. The median time from first diagnosis to start of RBP therapy was 27 (range 4–120) months, the median number of previous treatments was 1 (range 1–2), the median duration of the last response before initiation of RBP therapy was 10 (range 1–70) months. Twenty two patients (88%) responded after at least two cycles of RBP with 1 sCR, 5 nCR, 8 VGPR and 8 PR. The RBP regimen achieved a rapid decrease of the M-protein, reaching the best response already after the first cycle in four (16%) patients and after the second cycle in further eight (32%) patients. The median time to first haematological response was 28 days, and median time to best response was 56 days. Due to increased haematological toxicity a dose reduction in most patients required in subsequent cycles of therapy. 19 patients discontinued the treatment prematurely of stem cell transplantation (n = 7), hematological toxicity (n = 7), non hematological toxicity (n = 3) or progress (n = 2). The median progression-free and overall survival was 22 and 38 months, respectively. Conclusions: RBP is a highly effective therapy for patients with relapsed or refractory MM. However, dose reduction is necessary in many patients because of haematological toxicity Disclosure: No conflict of interest disclosed.

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V666

Independent factors influencing the outcome after allogeneic stem cell transplantation in patients with relapsed multiple myeloma: A retrospective single-center analysis in 98 patients Mielke S.1, Schemmel L.-K.1, Schreder M.1, Grigoleit G.-U.1, Rasche L.1, Bönig H.1, Stuhler G.1, Einsele H.1, Knop S.1 Universitätsklinikum Würzburg, Med. Klinik und Poliklinik II, Würzburg, Germany 1

Introduction: Standard treatment for fit patients with multiple myeloma (MM) includes induction therapy followed by autologous stem cell transplantation (SCT) providing substantial treatment-free intervals. With the vast majority of patients facing relapse or progression allogeneic SCT may be regarded as one treatment option aiming for long-term MM control by allo-immuno-surveillance. Patients: We have retrospectively analyzed all patients with MM relapsed after prior autoSCT receiving an alloSCT between 03/2005 and 12/2013 at our center. The data base was closed 6/2014. Patients with up front auto-allo or second allo SCT were excluded. Results: In total, 98 patients with a median age of 56 (36–68) years were assessable for final analysis with a median follow-up of 192 (3–2573) days from alloSCT; 18 had high risk cytogenetics with del17p and/or translocation (4;14) while 37 had extramedullary MM (eMM). Sixty-four had up to 3 and 34 had 4 to 9 treatment lines prior to alloSCT. Remission before alloSCT was CR in 4, VGPR in 12, PR in 37, SD in 25 and PD in 20. The majority of patients received RIC and matched PBMCs. Twenty-one patients received a primary “post-transplant treatment strategy” (PTTS). Overall 1- and 5-year survival (OS) was 36% and 22%. One and 4-year progression-free survival (PFS) was 24% and 7%. Non-relapse mortality was 16% at day 100 and 27% at one year (PD excluded as competing risk). Cumulative incidence of GvHD was 23% for grade II-IV aGvHD at day100 and 60% for cGvHD at three years (death excluded as competing risk). In multivariate analysis eMM (Hazard ratio 2.2), del17p (HR 2.2), 4–9 lines of therapy prior to alloSCT (HR 2.4) and Sorror-Score 4–6 (HR 2.3) evolved as a significant and negative impact on OS. PFS was negatively and significantly influenced by cGvHD (HR 2.9), eMM (HR 1.9), PD prior to alloSCT (HR 4.1) and Sorror Score 4–6 (HR 2.4). However, patients with absence of eMM, del17p, less than 4 prior lines of therapy and a Sorror score below 4 achieved a 5y-OS of 51%. Conclusions: OS in patients heavily pretreated or with higher Sorror Score was low with no beneficial effect of cGvHD and PTTS visible whilst del17p and eMM remained as independent negative prognosticators. Nevertheless, patients with less prior therapies and reduced disease burden in good condition may benefit from alloSCT. We are preparing to investigate the optimal treatment strategy for these patients in a prospective trial (DSMMXVI) including new agents. Disclosure: Stephan Mielke: Financing of Scientific Research: Celgene, Cellex; Other Financial Relationships: Celgene Stefan Knop: Financing of Scientific Research: Amgen, Takeda, Celgene; Other Financial Relationships: Takeda

Freier Vortrag

Translationale Forschung V667

Overcoming T cell exhaustion by targeting tumor suppressor p53 isoforms as a novel approach to improve T-cell based cancer immunotherapy Legscha K.J.1, Atunes E.1, Amann E.1,2, Theobald M.1,2,3, Echchannaoui H.1 III. Medizinische Klinik, Universitätsmedizin, Mainz, Germany, 2German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Frankfurt/Mainz, Germany, 3Research Center for Immunotherapy (FZI), Mainz, Germany 1

Introduction: Adoptive transfer of genetically modified T lymphocytes with tumor antigen-specific T-cell receptor (TCR) or chimeric antigen

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receptor (CAR) has proven efficacy in cancer immunotherapy. However, in many patients the overall benefit is still limited due to various tumor escape mechanisms. Cell damage and metabolic/hypoxic stress in the tumor microenvironment can lead to T cell exhaustion or senescence, a state characterized by proliferation arrest, cell dysfunction and change in phenotype. The tumor suppressor p53 is a master molecule in controlling the cell cycle and also regulates cell damage response and senescence. Here we show that genetic modification of p53 isoforms represents a novel approach to circumvent the tumor-derived immunosuppression of TILs and therefore could improve anti-tumor response after adoptive T cell transfer. Methods: Human T cells form healthy donors were retrovirally co-transduced with either p53β or Δ133p53 isoforms, together with a tumor antigen-specific TCR. Modified T cells were characterized for the expression of key activating/inhibitory molecules, homing markers and their proliferation capacity by flow cytometry. The effector functions (i.e. cytokine secretion and antigen-specific killing capacity) were assessed by Luminex immunoassay and long-term tumor colony-forming assay, respectively. Results: T cells overexpressing the Δ133p53 isoform revealed reduced levels of the inhibitory receptor Programmed cell death 1 (PD-1), and T-cell Ig and ITIM domain (TIGIT) expression and upregulation of the chemokine receptor CXCR3 while the p53β-modified cells showed increased expression of PD-1 and downregulation of CXCR3. Furthermore, after 15–18 weeks in culture, control T cells reached cellular senescence, characterized by poor proliferation, while, Δ133p53 cells remained proliferative and showed superior cytokine secretion levels. Importantly, Δ133p53 cells demonstrated an enhanced tumor-specific killing capacity. We are currently testing the therapeutic efficacy of p53 isoform-modified human T cells in mouse xenograft models. Conclusion: Genetic modulation of p53 isoforms in human T lymphocytes represents a novel approach to circumvent tumor-mediated T-cell replicative senescence and most importantly preserves long-term effector function of tumor reactive T cells. We believe that this new approach could improve current adoptive T-cell-based therapies. Disclosure: No conflict of interest disclosed. V668

Effects of the multi-kinase inhibitor DCC-2618 on proliferation and survival of neoplastic mast cells and comparison with other cell types Schneeweiss M.1,2, Peter B.1,2, Blatt K.1,2, Berger D.2, Stefanzl G.2, Hadzijusufovic E.1,2, Gleixner K.V.2, Valent P.1,2 Ludwig Boltzmann Cluster Oncology, Medizinische Universität Wien, Wien, Austria, 2Universitätsklinik für Innere Medizin I, Medizinische Universität Wien, Abteilung für Hämatologie und Hämostaseologie, Wien, Austria 1

Systemic mastocytosis (SM) is a myeloid neoplasm defined by excessive proliferation and accumulation of neoplastic mast cells (MC) in diverse organs. Most patients with SM express a D816V-mutated variant of KIT, which confers resistance against several tyrosine kinase inhibitors (TKI), including imatinib. Midostaurin (PKC412) is a TKI which is effective against KIT D816V. However so far, in most patients with advanced SM treated with PKC412, no continuous complete remission is obtained. One problem in advanced SM is that the associated AML or CMML component of the disease is often drug-resistant. We have investigated the effects of DCC-2618, a novel TKI directed against KIT, FLT3, FMS, and PDGFR, on proliferation and survival of neoplastic MC and other cell types. As assessed by 3H-thymidine-uptake, DCC-2618 inhibited proliferation of human MC leukemia cell lines, with lower IC50 values obtained with HMC1.1 cells (7.5–25 nM) and ROSAKIT WT cells (50–100 nM) compared to KIT D816V+ HMC-1.2 cells (100–500 nM) and ROSAKIT D816V cells (100–500 nM). In addition, DCC-2618 produced growth inhibition in the multi-resistant MCL line MCPV-1 (IC50: 50–500 nM). We were also able to show that DCC-2618 induces apoptosis in HMC-1 cells and ROSA cells, and less effectively in MCPV-1 cells. DCC-2618 was found to inhibit phosphorylation of KIT in all MC cell lines tested. In a next step we explored the effects of DCC-2618 on growth of other leukemias and on vascular en-

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dothelial cells. In these experiments, we were able to show that DCC-2618 inhibits the proliferation of the FIP1L1-PDGFRA+ eosinophilic leukemia cell line EOL-1 (0.1–1.0 nM) and the FLT3 ITD-mutated AML cell lines MV-411 and MOLM-13 (IC50: 50–250 nM). In these cells, DCC-2618 was also found to induce apoptosis as defined by light microscopy. Moreover, DCC-2618 was found to inhibit the growth of cultured human vascular endothelial cells. DCC-2618 did not inhibit the proliferation of the immature AML cell line KG1 and the monoblastic cell line U937, but was found to block growth and proliferation in primary leukemic monocytes in patients with AML-M4 and CMML. Together, our data show that DCC2618 is a potent multi-targeted TKI that counteracts growth of neoplastic cells in various myeloid leukemias in vitro. Whether DCC-2618 is also able to act anti-neoplastic on leukemic cells in vivo in patients with advanced SM, SM-AML, SM-CMML, or AML/CMML without SM remains to be determined in clinical trials. Disclosure: Mathias Schneeweiss: No conflict of interest disclosed. Peter Valent: Expert Testimony: Research Support von Deciphera Pharmaceuticals V669

Targeting NOTCH2 in human solid tumors in vitro and in a melanoma xenotransplant mouse model by aspergillus mycotoxin Hubmann R.1, Sieghart W.2, Schnabl S.2, Araghi M.2, Hilgarth M.2, Reiter M.2, Ponath E.2, Demirtas D.2, Zielinski C.2, Jäger U.2, Shehata M.2 Medical University of Vienna, Comprehensive Cancer Center Vienna, Wien, Austria, 2Medical University of Vienna, Vienna, Austria 1

Introduction: The highly conserved NOTCH gene family (NOTCH1–4) encodes transmembrane receptors that regulate embryonic development and adult issue homeostasis. Deregulation of NOTCH2 signaling is implicated in a wide variety of human solid tumors including hepatocellular carcinoma (HCC), pancreas carcinoma, bladder cancer, medulloblastoma, glioblastoma, and melanoma. The current concept of targeting NOTCH is based on using γ-secretase inhibitors (GSI) to regulate the release of the active NOTCH intracellular domain. However, the clinical outcome of GSI remains unsatisfactory. Methods: We analyzed the expression of nuclear NOTCH2 in several solid tumor cell lines and evaluated their sensitivity to GSI and to the recently identified NOTCH2 transactivation inhibitor gliotoxin, an Aspergillus derived mycotoxin (Hubmann et al., BJH2013). We also investigated the in vivo effect of gliotoxin in a melanoma xenograft model. Results: EMSA, as a surrogate method for the detection of nuclear NOTCH activity, showed that cell lines derived from melanoma (518A2), HCC (Hep3B, SNU398), and pancreas carcinoma (PANC1) express high amounts of DNA-bound NOTCH2 complexes. Gliotoxin efficiently targeted nuclear NOTCH2 and induced apoptosis in these cells whereas GSI were ineffective. The GSI resistance might be attributed to the expression of truncated NOTCH2 forms which do not depend on γ-secretase for processing and function.The induction of apoptosis by gliotoxin correlated with the nuclear NOTCH2 activity and was not associated with effects on NFκB signaling, and the redox status of the treated cells. The specificity of gliotoxin was demonstrated in the well differentiated nuclear NOTCH2 negative HCC cell line Huh7, which was found to be resistant to gliotoxin treatment in vitro.In xenotransplanted human 518A2 melanomas, a single day dosing schedule of gliotoxin was well tolerated and the mice did not show any study limiting side effects. Gliotoxin significantly decreased the mean tumor volume in early stage (83 mm3 vs 115 mm3, P = 0.008) as well as in late stage (218 mm3 vs 576 mm3, P = 0.005) tumors. Conclusion: The data show that targeting NOTCH2 by gliotoxin is a promising therapeutic approach in NOTCH2 associated neoplasias particularly those which do not respond to GSI treatment. We also show that Gliotoxin has a remarkable antitumor effect in vivo within a physiologically relevant and well tolerated concentration.

Abstracts

Disclosure: Rainer Hubmann: Honoraria: Gliotoxin for the treatment of NOTCH2 associated neoplasias. Medhat Shehata: Honoraria: Gliotoxin for the treatment of NOTCH2 associated neoplasias. V670

Phase 1 multicenter, open-label study to establish the maximum tolerated dose (MTD) of two administration schedules of E7389 (eribulin) liposomal formulation in patients (pts) with solid tumors Zubairi I.1, Dean E.2, Molife L.R.3, Lopez J.3, Ranson M.4, El-Khouly F.5, Savulsky C.6, Reyderman L.7, Jia Y.7, Hutton E.6, Morrison R.1,8, Sweeting L.1, Greystoke A.9, Barriuso J.2, Kristeleit R.5, Evans T.R.1,8 Beatson West of Scotland Cancer Centre, Glasgow, United Kingdom, 2The Christie NHS Foundation Trust, Manchester, United Kingdom, 3The Royal Marsden Hospital NHS Foundation Trust and The Institute of Cancer Research, London, United Kingdom, 4The Christie NHS Foundation Trust (retired), Manchester, United Kingdom, 5UCL Cancer Institute, London, United Kingdom, 6 Eisai Ltd, Hatfield, United Kingdom, 7Eisai Inc, Woodcliff Lake, United States, 8 University of Glasgow, Glasgow, United Kingdom, 9Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom 1

Background: Eribulin, a microtubule dynamics inhibitor, is approved in the US for the treatment of pts with metastatic breast cancer who have previously received ≥2 chemotherapeutic regimens for metastatic disease including an anthracycline and a taxane in the adjuvant/metastatic setting. Preclinical evidence suggested improved efficacy and lower toxicity with a liposomal formulation of eribulin (eribulin-LF). The primary objective of this study was to determine the MTD and dosing schedule for eribulin-LF. Secondary objectives included determining safety, pharmacokinetics (PK), and efficacy. Methods: Using a conventional 3 + 3 study design, eligible pts received escalating doses (1.0 to 3.5 mg/m2) of eribulin-LF (60-min, i.v. infusion) on day (D) 1 of a 21-day cycle (schedule [S] 1) or on D1 and D15 of a 28day cycle (S2). An expansion phase confirmed the safety of eribulin-LF in pts with endometrial, ovarian, or HER2-negative breast cancer. Results: Of 58 pts (S1: n = 20; S2: n = 38): 65% were female, median age was 62 yrs (range 33–75), 65% had an ECOG PS of 1, and 10 pts had metastatic breast cancer. Dose-limiting toxicities were: S1, grade (G) 4 hypophosphatemia (n = 1), G4 increased ALT/AST (n = 1), both at eribulin-LF 1.5 mg/m2; S2, G4 febrile neutropenia (n = 1; 1.5 mg/m2); G3 increased ALT/AST (n = 1), and G4 neutropenia (n = 1), both at 2 mg/m2. MTD was 1.4 mg/m2 in S1 and 1.5 mg/m2 in S2; the latter dose and schedule were used in the expansion phase (n = 23). Treatment-emergent adverse events (TEAEs) included peripheral neuropathy (S1: 35%, S2: 8%) and neutropenia (S1: 20%, S2: 34%). TEAEs of G3 or higher occurred in 55% (S1) and 39% (S2). Eribulin-LF exhibited single-phase PK, and was quantifiable ~18 days postdose. Exposure was dose-dependent and t1/2 was ~35 hrs. Promising clinical activity was observed in pts with breast cancer (5/10 partial responses, 3/10 stable disease). Conclusions: In this phase 1 study, eribulin-LF was well tolerated, with promising activity in pts with breast cancer. ClinicalTrials.gov identifier: NCT01945710 Funding Source: Eisai Inc. Disclosure: Ishtiaq Zubairi: Stock Ownership: Roche, GSK Thomas Evans: Advisory Role: Bristol-Myers Squibb, Baxter, Karus Therapeutics, Eisai; Financing of Scientific Research: Bristol-Myers Squibb; Expert Testimony: AstraZeneca, Celgene, Eisai, GSK, Roche, Basilea, e-Therapeutics, Immunocore, Vertex, Merck, Bristol-Myers Squibb, Daiichi

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Dual Fc-optimization of therapeutic antibodies for improved lymphoma therapy – enhanced complement-dependent and cell-mediated cytotoxicity by combined glyco- and proteinengineering Roßkopf S.1, Wirth T.1, Lutz S.1, Gramatzki M.1, Kellner C.1, Peipp M.1 Christian Albrechts University of Kiel, 2nd Department of Medicine, Division of Stem Cell Transplantation and Immunotherapy, Kiel, Germany 1

Introduction: Clinical data and animal models demonstrated the importance of Fc mediated effector functions including CDC and ADCC for the therapeutic activity of clinically approved monoclonal antibodies. Thus, to improve the efficacy of antibodies, current engineering strategies focus on modifying the Fc domain to selectively improve the interaction with either Fcγ receptors or complement to enhance ADCC and CDC, respectively. Improved antibody variants are designed either by protein-engineering or by glyco-engineering. However, whether both ADCC and CDC could be improved simultaneously by combining both technologies has to be determined. Methods: An Fc variant with improved C1q binding affinity (EFTAE-Fc) was introduced in the background of CD19 and CD20 antibodies. Fucosylated and afucosylated antibodies were expressed in CHO and Lec13 cells, respectively, and purified by affinity chromatography. Binding characteristics were analyzed by flow cytometry. The ability of the antibody variants to trigger CDC or ADCC in vitro was compared in 51Cr release assays using human plasma or mononuclear cells. Results: Two sets of antibodies targeting CD19 or CD20 were generated: native IgG1, their afucosylated variants, as well as fucosylated and afucosylated antibodies carrying EFTAE-Fc. Whereas native CD20 antibodies killed lymphoma cells by ADCC and CDC, native CD19 antibodies were not efficient. Interestingly, CD19 antibodies were enabled to mediate CDC by introduction of EFTAE-Fc. This mutation also enhanced CDC of CD20 antibodies, especially of target cells resistant to complement lysis triggered by native IgG1. In contrast, both afucosylated CD19 and CD20 antibodies had a higher ADCC capacity than their native counterparts. The higher ADCC activity achieved by glyco-engineering was found irrespective of the introduction of the EFTAE mutation, which selectively enhanced CDC. Importantly, double engineered variants without fucose and carrying the EFTAE mutation had both a higher CDC as well as ADCC activity in comparison with native IgG1 antibodies. Conclusion: Combining glyco- and protein-engineering technologies offer the opportunity to simultaneously enhance ADCC and CDC activities of therapeutic antibodies. This strategy may be particularly attractive to improve antibodies, such as CD19 antibodies with weak effector functions as native IgG1 antibodies, and may allow designing antibodies with tailor-made effector functions for improved lymphoma therapy. Disclosure: No conflict of interest disclosed. V672

Improved recovery of CD45RAnegCD8pos T lymphocytes and Induction of CD8-mediated antiviral reactivity following positive selection versus depletion of CD45RApos cells using CD45RA MicroBeads Teschner D.1, Kahlert L.1, Frey M.1, Khan S.A.1, Wagner E.M.1, Theobald M.1, Herr W.2, Hartwig U.F.1 Universitätsmedizin der Johannes Gutenberg-Universität Mainz, III. Med. Klinik & Poliklinik – Hämatologie, Internistische Onkologie & Pneumologie, Mainz, Germany, 2Universitätsklinikum Regensburg, Medzinische Klinik 3 Hämatologie und Internistische Onkologie, Regensburg, Germany 1

Introduction: Donor lymphocyte infusion (DLI) post allogeneic stem cell transplantation was shown to improve adoptive immunity to common infectious pathogens (CIP). However, alloreactive precursors are found particularly within naive CD45RA+CD4+ T cells as compared to memory, CD45RA- T cells.1 Moreover, previous studies revealed low numbers of CD8+ cells after CD45RA+ depletion.1 In this study we thus compared

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purity, recovery and anti-viral/fungal responses of CD45RA- T cells obtained either by depletion or positive selection of CD45RA+ cells followed by CD8-selection to improve immunity to CIP while reducing potential graft-versus-host reactivity. Methods: CD45RA+ T cells from HLA-A2+ PBMC of healthy donors (n = 5) were first removed either by depletion or positive selection using CD45RA MicroBeads. The CD45RA- subset was then further enriched for CD45RA-CD8+ cells. All fractions (i.e. unseparated, CD45RA-, CD45RACD8+) were analyzed by phenotypic and functional assays. Results: CD45RA+ cells were effectively removed by both depletion or positive selection (median 2.8 log depletion) and an 1,7fold increase of CD45RA-CD4+ T cells compared to untreated PBMC was observed by both approaches. However, significantly more CD45RA-CD8+ T cells were obtained following elimination of CD45RA+ cells by positive selection (36,8% vs. 3,0%). Accordingly, subsequent CD8 enrichment was 10fold higher when compared to the amount of CD8+ cells obtained after CD45RA depletion (4,7% vs. 0,5%) while purity was comparable (88% vs. 90%). Both strategies resulted in nearly complete elimination of NK cells and B cells. Interestingly, CD15+ granulocytes were only enriched upon CD45RA+ depletion. In IFN-g ELISpot assays, persistent and primarily CD4-mediated reactivity to candida, and aspergillus antigens was observed in the CD45RA- subset after both selection strategies. In contrast, responses to cytomegalovirus and Epstein-Barr virus were mainly CD8-driven and thus elevated in the CD45RA-CD8+ subset. Studies on residual alloreactivity within the CD45RA- fractions are in progress and will be reported. Conclusion: CD45RA depleted DLI contains memory T cells that show preserved reactivity to CIP. Removal of CD45RA+ T cells by positive selection leads to better recovery of CD45RA-CD8+ cells and might thus be an interesting approach to study modified DLI for improved adoptive immunity but reduced risk of GvHD. Reference: 1 Teschner D et al.: BMT. 2014.49:138–144. First and second author and the last two authors shared authorship Disclosure: Daniel Teschner*: Advisory Role: Pfizer Deutschland GmbH, MSD Sharp & Dohme GmbH; Financing of Scientific Research: Gilead Sciences GmbH, MSD Sharp & Dohme GmbH; Other Financial Relationships: Astellas Pharma GmbH , Gilead Sciences GmbH, Jazz Pharmaceuticals Germany, MSD Sharp & Dohme GmbH (Reisekostenerstattungen) Udo Hartwig°: No conflict of interest disclosed.

Plenarsitzung Best Abstracts V673

Target expression, preclinical activity and mechanism of action of EM801: a novel first-in-class IgG-based BCMA T-cell bispecific antibody for the treatment of multiple myeloma Seckinger A.1, Delgado J.A.2, Moreno L.2, Neuber B.1, Grab A.L.1, Lipp S.1, Merino J.2, Vu M.D.3, Strein K.3, Prosper F.2, Hundemer M.1, San Miguel J.2, Hose D.1, Paiva B.2 Universitätsklinikum Heidelberg, Medizinische Klinik V, Heidelberg, Germany, Clinica Universidad de Navarra, Department of Hematology and Immunology, Pamplona, Spain, 3EngMab AG, Wilen, Switzerland 1 2

Introduction: T-cell bispecific antibodies (TCBs) couple T-cells with antigens on tumor cells conferring T-cell mediated tumor cell death. Our aims were i) to assess expression of B-cell maturation antigen (BCMA) as potential target in multiple myeloma (MM); ii) to evaluate activity of EM801 BCMA-TCB; and iii) to delineate its mechanism of action. Methods: BCMA expression was investigated in plasma cells (PCs) from newly diagnosed (n = 630) and relapsed (n = 86) MM patients, as well as bone marrow (BM) subsets from healthy donors using DNA-microarrays, RNA-sequencing and flow cytometry. Crosslinking of MM- and T-cells

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was assessed using atomic force and electron scanning microscopy. T-cell activation was studied using flow cytometry and ELISA. Activity of EM801 in vitro was investigated in 43 BM samples from MM patients, in vivo activity was studied in a subcutaneous H929 myeloma cell line xenograft model in NOG (NOD/Shi-scid/IL-2Rγnull) mice reconstituted with human PBMCs. Results: BCMA expression was observed in malignant PCs from all 716 MM patients, 10/10 normal PCs and 5/5 plasmablasts while being absent in other subsets. Data were confirmed using flow cytometry (n = 43). BCMA is thus a potential TCB target in all previously untreated and relapsed MM patients including those with highly proliferative disease or high-risk features according to chromosomal aberrations or gene expression-based risk scores. EM801 concentration-dependently couples T-cells and PCs up to 7-fold of control (P < .001) resulting in higher T-cell activation with significantly increased expression of CD69, CD25 (both P < .001) and HLADR (P = 0.001) in both CD4+ and CD8+ T-cells, and secretion of interferon-γ, granzyme B (both P < .001), and perforin (P < .01).EM801 at 10pM-30nM induced significant cell death in PCs of 24/31 (77%) newly diagnosed and 10/12 (83%) relapsed MM patients including samples from heavily pretreated patients. In vivo, complete tumor regression was observed in 6/9 mice while an unspecific TCB did not show any efficacy. Toxicity on BM subsets was negligible. Conclusions: BCMA is an excellent target in newly diagnosed and relapsed MM patients. Yet increasing the binding strength between MMand T-cells by EM801 overcomes inhibitory mechanisms resulting in high single agent activity in vitro and in vivo. Given the elegant application schedule (i.v. or s.c. 1x/week), EM801 is perfectly suited for non-cross-resistant single agent, combination, or long-term treatment.

Methods: Data on cancer patients with influenza were collected from medical records from 8 university hospitals using a predefined short questionnaire. Patients (outpatients and inpatients) with active cancer or a history of cancer were included from November 2014 to June 2015. Results: Two-hundred and three patients with haematological malignancies and solid tumours were identified; 109 patients (54%) had active disease. Influenza A was detected in 155 (76%) and influenza B in 46 patients (23%) by PCR from respiratory samples (unknown subtype in 2 patients). Symptoms at clinical presentation included upper-respiratory tract infection in 55 (27%), influenza-like illness in 82 (40%) and pneumonia in 57 (28%). Another 10 patients developed pneumonia later on. Fifty-nine percent (116/195) received oseltamivir. Superinfections occurred in 37 (18%) and ICU-treatment was required in 26 (13%). Sixteen patients (8%) died. Multivariate analysis revealed superinfection (HR 3.4 95%, confidence interval (CI) 1.09–10.6) and duration from onset of symptoms to diagnosis (HR 1.1 95%, CI 1.01–1.2) as independent prognostic factors. Conclusion: Influenza-infections have a high rate of pneumonia and a high mortality-rate in patients with cancer. Early diagnosis should be attempted. Superinfections need to be addressed as soon as possible since this is the most dangerous complication. Disclosure: Daniel Teschner: Advisory Role: Pfizer Deutschland GmbH, MSD Sharp & Dohme GmbH; Financing of Scientific Research: Gilead Sciences GmbH, MSD Sharp & Dohme GmbH; Other Financial Relationships: Astellas Pharma GmbH , Gilead Sciences GmbH, Jazz Pharmaceuticals Germany, MSD Sharp & Dohme GmbH (Reisekostenerstattung) Marie von Lilienfeld- Toal: Advisory Role: Consultant of advisory board for MSD; Financing of Scientific Research: Honoraria, travel grant from MSD, Celgene, Gilead, Janssen Cilag and Astellas; Expert Testimony: research grant from MSD and Celgene

Disclosure: Anja Seckinger: Expert Testimony: EngMab AG Bruno Paiva: Expert Testimony: EngMab AG V674

Influenza virus infections in patients with malignancies: Characteristics and outcome of the spring season 2015 - final analysis by the infectious diseases working party (AGIHO) of the German Society for Hematology and Medical Oncology (DGHO) Teschner D.1, Hermann B.2, Lehners N.3, Brodhun M.4, Hochhaus A.4, Boden K.5, Kochanek M.6, Meckel K.4, Mayer K.7, Rachow T.4, Rieger C.8, Schalk E.9, Schmeier-Jürchott A.1, Weber T.10, Schlattmann P.4, von Lilienfeld-Toal M.4, AGIHO III. Medizinische Klinik, Universitätsklinikum Mainz, Mainz, Germany, Universitätsklinikum Jena, Jena, Germany, 3Medizinische Klinik V der Ruprecht-Karls-Universität, Heidelberg, Germany, 4Klinik für Innere Medizin II, Universitätsklinikum, Jena, Germany, 5Universitätsklinikum Jena, Institut für Klinische Chemie, Jena, Germany, 6Universitätsklinikum Köln, Klinik I für Innere Medizin, Köln, Germany, 7Universitätsklinikum Bonn, Medizinische Klinik und Poliklinik III Abteilung für Onkologie, Hämatologie, Immunonkologie und Rheumatologie, Bonn, Germany, 8Ludwig-Maximilians-Universität München, Medizinische Klinik und Poliklinik III, München, Germany, 9University Hospital Otto-von-Guericke University Magdeburg, Medical Centre, Department of Haematology and Oncology, Magdeburg, Magdeburg, Germany, 10 Universitätsklinikum Halle, Klinik für Innere Medizin IV -Hämatologie und Onkologie-, Halle, Germany 1 2

Introduction: Infections are the main cause of treatment-related mortality in cancer patients. Whereas bacterial and fungal infections are common and well-known, infections by community acquired viruses such as influenza have been neglected in the past. Furthermore, most studies that investigate infections with influenza focus on patients with hematological malignancies and hematologic stem cell transplantation, but information on other cancers are scarce. We sought to understand the clinical epidemiology and outcome of cancer patients who had documented influenza in the 2015 season in order to identify patients at risk for severe course of infection and mortality.

Abstracts

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V675

Stopping tyrosine kinase inhibitors in a large cohort of European chronic myeloid leukemia (CML) patients: results of the EURO-SKI trial Saußele S.1, Mahon F.-X.2, Guilhot J.3, Dengler J.4, Waller C.T.5, Brümmendorf T.H.6, Herbst R.7, Burchert A.8, Goebeler M.-E.9, Freunek G.10, Prümmer O.11, Jost P.J.12, Prange-Krex G.13, Illmer T.14, Janzen V.15, Schafhausen P.16, Klausmann M.17, Kiani A.18, Eckert R.19, de Wit M.20, Janssen C.21, Müller M.C.22, Hochhaus A.23, Richter J.24, Pfirrmann M.25, EURO-SKI Investigators Medizinische Fakultät Mannheim, Universität Heidelberg, III. Medizinische Klinik, Mannheim, Germany, 2University of Bordeaux, Bergonié Cancer Institute, INSERM Unit 916, Bordeaux, France, 3CHU de Poitiers, Inserm CIC 1402, Poitiers, France, 4Onkologische Schwerpunktpraxis Heilbronn, Heilbronn, Germany, 5Universitätsklinikum Freiburg, Klinik für Innere Medizin I, Freiburg, Germany, 6Uniklinik RWTH Aachen, Medizinische Klinik IV, Aachen, Germany, 7 Klinikum Chemnitz, Klinik für Innere Medizin III, Chemnitz, Germany, 8 Universitätsklinikum Gießen und Marburg, Standort Marburg, Philipps Universität, Klinik für Hämatologie/Onkologie und Immunologie, Marburg, Germany, 9Universitätsklinikum Würzburg, Medizinische Klinik und Poliklinik II, Würzburg, Germany, 10Medizinischen Versorgungszentrums am Klinikum St. Elisabeth, Fachbereich Onkologie und Hämatologie, Straubing, Germany, 11 Klinikum Kempten, Klinik für Hämatologie, Onkologie und Palliativmedizin, Kempten, Germany, 12Klinikum rechts der Isar, III. Medizinischen Klinik, München, Germany, 13Fachärztliche Gemeinschaftspraxis für Innere Medizin, Hämatologie und Onkologie, Dresden, Germany, 14Fachärztliche Gemeinschaftspraxis, Schwerpunkt Hämatologie und Onkologie, Dresden, Germany, 15Universitätsklinikum Bonn, Medizinische Klinik und Poliklinik III, Bonn, Germany, 16Universitätsklinikum Hamburg Eppendorf, II. Medizinische Klinik und Poliklinik, Hamburg, Germany, 17Onkologische Gemeinschaftspraxis Drs. Klausmann, Aschaffenburg, Germany, 18Klinikum Bayreuth, Klinik für Onkologie und Hämatologie, Bayreuth, Germany, 19Onkologische Schwerpunktpraxis, Esslingen, Germany, 20Vivantes Klinikum Neukölln, Klinik für Hämatologie, Onkologie und Palliativmedizin, Berlin, Germany, 21Onkologie Unter Ems, Leer, Germany, 22IHO Institute for Hematology and Oncology, Mannheim, Germany, 23Universitätsklinikum Jena, Abteilung Hämatologie/ Onkologie, Klinik für Innere Medizin II, Jena, Germany, 24Skåne University Hospital, Department of Hematology, Oncology and Radiation Physics, Lund, Sweden, 25Ludwig-Maximilians-Universität, Institut für Medizinische Informationsverarbeitung, Biometrie und Epidemiologie, München, Germany 1

In several studies, the possibility of treatment-free remission in CML could be shown (Mahon 2010). However, prognostic factors and commonly acknowledged procedures for a successful treatment stop in CML are not yet defined. The EURO-SKI study (European stop TKI (tyrosine kinase inhibitors) study) was set up to estimate the proportion of patients in major molecular remission (MMR) at 6 months (m) after stopping TKIs. Secondary endpoints included the identification of prognostic factors and safety aspects. Adult CML patients in chronic phase (CP) CML on TKI treatment for at least 3 years and in MR4 (BCR-ABL according to international scale < 0.01%) for at least one year were eligible. Patients with prior TKI failure were excluded. Here, we report results of all patients, with a minimum follow-up of 6m, additionally German patients were analyzed for safety aspects. 868 patients in CP CML from 11 countries were included, 772 patients were eligible, 168 from Germany; 46.6% were female. Median age at stop was 60.3 years (range 19 to 89). 10% were at high risk according to the EUTOS Score. First-line TKI was imatinib in 94%, dasatinib in 2% and nilotinib in 4%. 115 patients switched to second-line TKI due to intolerance. Time from diagnosis of CML to first day of stopping TKI varied from 36 to 270m, median time was 92m. Median duration of TKI treatment was 91m (range 36–170m) and median duration of MR4 prior to TKI stop was 56m (12–160m). 717 patients had assessable molecular data for the estimation of molecular relapse-free survival (mRFS). Of these patients, 331 lost MMR, 4 died in remission and 381 are still in MMR at last-follow-up (range 1–36 m). This resulted in a mRFS of 62% (95% confidence interval (CI): 58–65%) at 6 m and 56% (CI: 52–59%) at 12 m. In Germany, 84 patients lost MMR. 14 of them were diagnosed with BCRABL levels >1% (range 1–16%). In 6 of these cases the rise up to 1% oc-

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curred between detection of loss of MMR and restart of TKI therapy. Nine of 72 patients had BCR-ABL levels between MR4 and MMR at month 12. Of those, 4 relapsed later on. In the setting of the EURO-SKI study, high mRFS rates are achievable. German experience suggests that logistics for restart of TKI treatment have to be organized more stringent as BCR-ABL levels >1% were observed. In patients with undulating PCR results one year after TKI stop, PCR control intervals of 3 months have to be discussed. Disclosure: Susanne Saußele: Financing of Scientific Research: Novartis, BMS, Pfizer, Ariad; Expert Testimony: Novartis, BMS Markus Pfirrmann: No conflict of interest disclosed. V676

EUCROSS: A European phase II trial to evaluate efficacy and safety of crizotinib treatment in advanced adenocarcinoma of the lung harbouring ROS1 translocations – Preliminary results Michels S.1, Thurat M.1, Schmalz P.2, Pereira E.3, Scheffler M.1, Fischer R.1, Sebastian M.4, Abreu D.R.5, Carcereny E.6, Corral J.J.7, Felip E.8, Grohé C.9, Insa A.10, Thomas M.11, Reck M.12, Rothschild S.13, Brandes V.1, Nogova L.1, Merkelbach-Bruse S.14, Massuti B.15, Büttner R.14, Rosell R.16, Wolf J.1, Lung Cancer Group Cologne and Spanish Lung Cancer Group Uniklinik Köln, Klinik I für Innere Medizin, Lung Cancer Group Cologne, Köln, Germany, 2Universität Köln, Zentrum für klinische Studien, Köln, Germany, 3 Spanish Lung Cancer Group, Barcelona, Spain, 4Uniklinik Frankfurt, Frankfurt, Germany, 5Hospital Universitario Materno-Infantil De Canarias, Spanish Lung Cancer Group, Las Palmas, Spain, 6Hospital Universitari Germans Trias i Pujol, Spanish Lung Cancer Group, Barcelona, Spain, 7Hospital Universitario Virgen del Rocío, Spanish Lung Cancer Group, Sevilla, Spain, 8Val d’Hebron Hospital Barcelona, Spanish Lung Cancer Group, Barcelona, Spain, 9Evangelische Lungenklinik Berlin, Berlin, Germany, 10Hospital Clínico Universitario de Valencia, Spanish Lung Cancer Group, Valencia, Spain, 11Thoraxklinik im Universitätsklinikum Heidelberg, Internistische Onkologie der Thoraxtumoren, Translational Lung Research Center, Heidelberg, Germany, 12Lungen Clinic Grosshansdorf, Grosshansdorf, Germany, 13Universitätsspital Basel, Basel, Switzerland, 14Uniklinik Köln, Institut für Pathologie, Lung Cancer Group Cologne, Köln, Germany, 15University Hospital Miguel Hernandez Alicante, Spanisch Lung Cancer Group, Alicante, Spain, 16Catalan Institute of Oncology, Spanish Lung Cancer Group, Badalona, Spain 1

Introduction: Translocations of ROS1 account for 1–2% of all oncogenic events in lung adenocarcinomas and predominantly occur in non-smokers. The ROS1 inhibitor crizotinib has shown to be highly effective in this patient subgroup in the PROFILE1001 study. EUCROSS is an investigator-initiated trial of the Lung Cancer Group Cologne and the Spanish Lung Cancer Group and the first prospective European trial to evaluate the efficacy of crizotinib in ROS1 positive NSCLCs. Here, we present preliminary data on efficacy and molecular characteristics. Methods: Patients with advanced lung adenocarcinomas harbouring ROS1 translocations as confirmed by central FISH analysis were eligible for the trial, irrespectively of the number of prior treatment lines. Patients initially received treatment with crizotinib 250 mg BID. The primary endpoint of this phase II trial is the overall response rate (ORR). Among the secondary endpoints are PFS, OS and safety. All efficacy endpoints will be assessed by investigator’s RECIST analysis and IRB. The baseline tumor tissue is analysed by CAGE sequencing to identify the translocation partners of ROS1, to validate the FISH analysis and to identify additional biomarkers for prediction of response. At time of progression an optional fresh frozen rebiopsy is scheduled to identify mechanisms of resistance to crizotinib by MPS, WES or WGS. Results: In total, 33 patients have been enrolled within EUCROSS and sequencing results of 20 patients were available at the time of data cut-off for this abstract (January 2016). Twenty-five (N = 25) patients had sufficient response assessments for efficacy evaluation at time of data cut-off. The ORR was 68% (N = 17; 95% confidence interval [CI] 49.7 to 86.3) in the overall trial population and 85% (N = 17; 95% confidence interval [CI] 69.3 to 100.6]) in the population with confirmed ROS1 rearrangement by sequencing. Two patients (8%; confidence interval [CI] 0.0 to 18.6) exhibited primary progression on treatment and were ROS1 negative

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by sequencing. The most common fusion partners of ROS1 were CD74 (N = 11; 52%), EZR (N = 4; 19%) and SCL34A2 (N = 4; 19%). Conclusions: Crizotinib is highly effective and safe in ROS1 translocated NSCLC patients. Although, the number of patients with ROS1 positivity by sequencing was low at the time of data-cut-off, the high ORR and the lack of primary progressing patients argues in favor for DNA-sequencing as the gold-standard for identification of true rearrangements. Disclosure: Sebastian Michels: Advisory Role: Pfizer, Novartis, Boehringer Ingelheim; Financing of Scientific Research: Pfizer, Novartis, Boehringer Ingelheim; Expert Testimony: Pfizer Jürgen Wolf: Advisory Role: Pfizer, Novartis, Boehringer Ingelheim, Astra Zeneca; Financing of Scientific Research: Pfizer, Novartis, Boehringer Ingelheim, Astra Zeneca; Expert Testimony: Pfizer V677

The role of CBX proteins in human benign and malignant hematopoiesis Jung J.1, Schepers H.2, Lazare S.1, Buisman S.1, Weersing E.1, Dethmers B.1, Klauke K.1, Bystrykh L.1, de Haan G.1 University Medical Center Groningen, European Research Institute for the Biology of Ageing (ERIBA), Groningen, Netherlands, 2University Medical Center Groningen, Experimental Hematology, Groningen, Netherlands 1

Introduction: Hematopoietic stem cells (HSCs) need to properly balance self-renewal and differentiation, in order to prevent malignant transformation or HSC exhaustion. We recently showed that the presence of specific Cbx proteins in the Polycomb Repressive Complex 1 (PRC1) plays a crucial role in balancing self-renewal and differentiation of murine HSCs. Whereas Cbx7 induces self-renewal, Cbx4 and Cbx8 instead induce differentiation. In this project, we investigate whether this role is evolutionary conserved in human CD34+ stem and progenitor cells (HSPCs), we aim to identify genes which are controlled by CBX7 and CBX8 containing PRC1 in human CD34+ HSPCs and try to explore the role of CBX7 in malignant hematopoiesis. Methods: We overexpressed CBX2,-4,-6,-7 and CBX8 in human CD34+ HSPCs. We assessed functional consequences by measuring cobblestone area-forming cell frequencies (CAFC), colony-forming unit (CFU) frequencies and stem cell xenotransplantation studies in immunodeficient NSG mice. For identifying genes, which are controlled by CBX7 or CBX8 containing PRC1, we are actually performing RNA- and Chip-seq in CD34+ HSPCs. For exploring the role of CBX7 in malignant hematopoiesis, we used a short-hairpin approach in human leukemic cell lines. Results: Overexpression of both, CBX7 and CBX8, in huCD34+ HSPCs led to a significant increase of week 5 CAFC frequencies (fold increase: 9,62 for CBX7; 4,92 for CBX8) and higher CFU frequencies after 14 days (fold increase: 1,32 for CBX7; 1,41 for CBX8). In contrast, the overexpression of CBX4 and CBX2 led to equal or decreased number of CAFC and CFU frequencies. Transplantation of CBX7 overexpressing human CD34+ HSPCs in sublethally irradiated NSG mice led to higher engraftment in comparison to the empty vector control (p < 0,05). Even after one week of in vitro culture, transplantation of CBX7-overexpressing cells resulted in significant higher engraftment levels (p < 0,05). To explore whether CBX7 is also required for self-renewal of leukemic cells, we knocked down CBX7 mRNA by short hairpins in the human promyelocytic leukemia cell line HL60 and in the CML cell line K562, and found that this induced markedly decreased proliferation. Conclusions: Together, our study indicates that CBX7 and CBX8 plays a role in maintenance or self-renewal of human HSC, while their close homologs CBX2 and -4 induce differentiation. Furthermore, our data suggest that CBX7 is required for self-renewal of leukemic cells. Disclosure: No conflict of interest disclosed.

V678

Outcome of allogeneic stem cell recipients admitted to the ICU – a single center analysis of 942 patients Lueck C.1, Stadler M.1, Koenecke C.1, Hoeper M.M.2, Dammann E.1, Schneider A.S.3, Kielstein J.T.4, Ganser A.1, Eder M.1, Beutel G.1 Medizinische Hochschule Hannover, Hämatologie, Hämostaseologie, Onkologie und Stammzelltransplantation, Hannover, Germany, 2Medizinische Hochschule Hannover, Pneumologie, German Center for Lung Research (DZL), Hannover, Germany, 3Medizinische Hochschule Hannover, Gastroenterologie, Hepatologie und Endokrinologie, Hannover, Germany, 4Medizinische Hochschule Hannover, Nieren- und Hochdruckerkrankungen, Hannover, Germany 1

Introduction: In 10%-40% of allogeneic hematopoietic stem cell transplant (HSCT) recipients life-threatening complications lead to intensive care unit (ICU) admission. Despite improved ICU treatments and use of reduced intensity regimens, mortality in critically ill HSCT recipients remains high, in particular in case of acute respiratory failure. However, analyses of large cohorts of critically ill HSCT recipients are rare. Methods: Between January 2000 and December 2013 942 patients underwent HSCT at our institution. Of those, 330 patients were at least once admitted to the ICU and included in this analysis. To identify differences over time the study group was split into two periods, 2000 - 2006 (n = 125) and 2007 - 2013 (n = 205). Results: Reasons for ICU admission were respiratory failure in 32% (n = 105), severe sepsis in 21% (n = 70), and cardiac events in 16% (n = 53). Another 15% (n = 50) were admitted for operative procedures. Other reasons like bleeding, impaired consciousness, and acute renal failure occurred in 13% (n = 42). Graft-versus-host disease accounted for only 3% (n = 10). Short-term mortality was 62%, 53% and 32% for patients admitted with respiratory failure, sepsis and cardiac events (p < 0.01). Longitudinal analysis revealed no difference in clinical reasons leading to ICU admission over time (p = 0.1). However, as the most important finding of this analysis the percentage of respiratory failure as a leading cause for ICU admission dropped from 40% to 27% (p < 0.01). Overall ICU and hospital mortality decreased from 52% to 37% (p < 0.01) and from 72% to 53% (p = 0.001), respectively. After hospital discharge 1- and 2-year-mortality rates declined not significantly from 51% to 27% and from 54% to 42% (p = 0.3). Remarkably, overall survival after hospital discharge was comparable between ICU and non-ICU patients (52% vs 60%, p = 0.08). Conclusion: To our knowledge this is the largest outcome analysis of critical ill HSCT recipients consecutively transplanted in a single center in Germany. This analysis identified respiratory failure, septic shock, and cardiac events as the major causes of ICU admission. The latter was associated with significantly better outcome. In general, short-term survival of critically ill HSCT recipients has improved over time. Long-term survival and mortality rates after hospital discharge were comparable regardless of any need for intensive care supporting the strategy of full code management strategy in HSCT patients. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium

Pathogenese und innovative Therapiekonzepte BCR-ABLnegativer myeloproliferativer Neoplasien V680

The role of non-driver mutations in myeloproliferative neoplasms Döhner K.1, Stegelmann F.1 Universitätsklinik Ulm, Klinik für Innere Medizin III, Ulm, Germany

Myeloproliferative neoplasms (MPN) are a group of diseases characterized by increased proliferation of erythroid, granulocytic and/or megakaryocytic cells. In the last decade, major progress has been made in un-

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derstanding the underlying molecular mechanisms, in particular for the three disease entities polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). Here, mutations in the genes JAK2, MPL or CALR occur in the vast majority of the patients (pts), and data from in vitro and in vivo studies showed that each of these mutations can induce the MPN phenotype with hyperactive JAK/STAT signaling as the common pathogenic basis. Based on these findings mutations in JAK2, MPL and CALR are considered as driver mutations in MPN. With the development of novel technologies (e.g. high-resolution genomic profiling and next-generation sequencing [NGS]), a growing list of additional mutations has been identified in MPN pts in recent years (e.g. ASXL1, SRSF2, EZH2, TET2, DNMT3A, IDH1/2). The affected genes belong to different functional categories encompassing e.g. epigenetic regulation, transcription or cell signaling acting via modification of effects mediated by the driver mutations. However, none of these gene alterations can cause a MPN phenotype; moreover, they occur frequently in other myeloid malignancies such as myelodysplastic syndrome or acute myeloid leukemia. Based on these features they are termed non-driver or important passenger mutations in MPN, with their exact biological function to be defined. Recently, Vannucchi et al. (Leukemia 2013) evaluated the clinical impact of non-driver mutations in 879 PMF pts. Here, ASXL1, SRSF2 and EZH2 mutations were independently associated with poor survival, and ASXL1 mutations remained significant in the context of the international prognostic Scoring System (IPPS) or the Dynamic IPSS (DIPSS)-plus model. In addition, IDH1/2, SRFS2 and ASXL1 mutations revealed as negative predictors for leukemia-free survival. In another study presented by Tefferi A et al. (ASH 2015, abstract #350, #354) a 27-gene NGS panel was used to analyze 494 MPN pts. Non-driver mutations occurred in >80% of PMF pts and in >40% of pts with PV or ET. The number of mutations and the presence of distinct ones were likely to predict clinical outcome. In this session, we will discuss the most recent data on the clinical relevance of non-driver mutations and their possible function in MPN pathogenesis. Disclosure: Konstanze Döhner: Advisory Role: Novartis Pharma GmbH; Financing of Scientific Research: Vortragshonorare Novartis Pharma GmbH Frank Stegelmann: Financing of Scientific Research: Vortragshonorare Novartis Pharma GmbH V682

Genetic predisposition to myeloproliferative neoplasms Cross N.C.P.1 University of Southampton, Wessex Regional Genetics Laboratory, Salisbury, United Kingdom 1

Evidence from epidemiological, familial and genetic studies indicate that common, low penetrance variants present in the general population contribute to the risk of developing MPN and also contribute to the phenotypic pleiotropy observed in these disorders. Although such variants are unlikely to be of direct clinical relevance, their identification may provide novel biological insights into the development of these disorders. For example, variants that predispose to MPN may confer a clonal advantage to mutant stem cells whereas variants that differentially predispose to PV or ET may influence differentiation decisions of megakaryocyte-erythroid progenitor cells. Several years ago, we and others identified a specific JAK2 haplotype, termed 46/1, that is strongly associated with acquisition of the JAK2 V617F mutation. The molecular basis for this association is not understood, but circumstantial evidence lends some support to this haplotype having a higher mutation rate (hypermutability hypothesis) and functional differences compared to other haplotypes (fertile ground hypothesis). In a recent genome wide association study (GWAS) we found that genetic variation at MECOM, TERT, JAK2 and HBS1L-MYB predisposes to JAK2-unmutated MPN. Furthermore, focused analysis of these four loci demonstrated that rs9376092 at HBS1L-MYB and the JAK2 46/1 haplotype specifically influence whether JAK2 V617F mutated cases present with PV or ET. To explore the possibility that variation at other genetic

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loci influence MPN phenotype, we undertook a two stage genome-wide association study of JAK2 V617F-mutated MPN. At stage 1, 556 ET and 556 PV patients with JAK2 V617F were genotyped using Illumina Human OmniExpressExome v1.2 BeadChips. At stage 2, significant SNPs from stage 1 were genotyped in four additional JAK2 V617F-positive ET/ PV cohorts from the UK (n = 180), Spain (n = 664), Italy (n = 547) and Germany (n = 74). We found that germline variation in the intergenic regions between HBS1L and MYB exerts the strongest influence on JAK2 V617F mutated MPN phenotype. In addition, variation between GFI1B and GTF3C5 predisposes to PV and in particular to cases with high JAK2 V617F mutation burden associated with 9p aUPD. Our findings indicate a complex interplay between somatic and germline genetics in MPN, and provides pointers towards the functional basis of the phenotypic pleiotropy of these disorders. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium Young investigators award V683

Naturally presented CML-associated HLA class II antigens elicit spontaneous CD4+ T-cell responses in CML patients Bilich T.1,2, Kowalewski D.J.1, Schemionek M.3, Kanz L.2, Salih H.R.2,4, Brümmendorf T.3, Vucinic V.5, Niederwieser D.5, Rammensee H.-G.1,6, Stevanovic S.1,6, Stickel J.S.2 Universität Tübingen, Abteilung für Immunologie, Tübingen, Germany, Universitätsklinik Tübingen, Hämatologie und Onkologie, Tübingen, Germany, 3 Universitätsklinikum RWTH Aachen, Abteilung für Hämatologie, Onkologie, Hämostaseologie und SCT, Aachen, Germany, 4Clinical Cooperation Unit Translational Immunology, German Cancer Consortium (DKTK), DKFZ partner site Tübingen, Tübingen, Germany, 5Universitätsklinikum Leipzig, Abteilung für Hämatologie und Onkologie, Leipzig, Germany, 6German Cancer Consortium (DKTK), DKFZ partner site Tübingen, Tübingen, Germany 1 2

While the discovery of BCR-ABL and the respective tyrosine kinase inhibitors (TKI) resulted in a significant prolongation of patient survival, there still is no curative treatment for chronic myeloid leukemia (CML) except for allogenic stem cell transplantation. The concept of T-cell based immunotherapy, such as for example with peptide-based vaccination, is a promising and low side effect approach in particular to eliminate residual leukemic cells, which might promote disease relapse after TKI discontinuation. As effective antigen-specific T-cell based cancer immunotherapy requires exact knowledge of tumor-associated epitopes that can act as rejection antigens, we have recent years developed in a mass spectrometry-based tech platform that allows for the direct identification of naturally presented tumor-associated HLA ligands in hematological malignancies including CML. Because of the important indirect and direct roles of CD4+ T-cells in anti-cancer immune responses, an optimal vaccine in our view requires the inclusion of HLA class II epitopes. In the present study we analyzed the HLA class II ligandome of primary CML cells (n = 15, 5,638 HLA ligands, 2,776 source proteins). Comparative ligandome profiling using a HLA class II database, which includes various normal tissues (n = 44, 13,783 HLA ligands, 7,758 source proteins including peripheral blood, bone marrow, kidney, liver, colon and bile duct), revealed 7 CML-associated antigens (50 HLA ligands) showing CML-exclusive representation in ≥25% of the CML patients. To evaluate the immunogenicity and specificity of these HLA class II CML-associated peptides, we performed IFNγ-ELISPOT assays after 12-day recall stimulation. A panel of 7 peptides was implemented for stimulation of PBMCs obtained from CML patients and healthy volunteers (HV). IFNγ-ELISPOT revealed peptide-specific immune recognition of 7 out of 7 (100%) CML-exclusive peptides in CML patients. The frequencies of the detected immune responses ranged from 17% (4/23) to 4% (1/23) of tested CML patient samples. These immune responses were mediated by functional CML patient derived CD4+ T-cells and were strictly CML-directed, as no

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immune response against CML-associated peptides was detected in HV (0/10). Taken together, these results are a first step towards validation and implementation of these newly defined CML-associated HLA class II antigens as targets for off-the-shelf peptide vaccination in CML patients. Disclosure: No conflict of interest disclosed. V684

Amplicon based next-generation-sequencing to quantify minimal residual disease in B-cell precursor acute lymphoblastic leukemia Koopmann J.1, Knecht H.1, Füllgrabe M.1, Pott C.1, Gökbuget N.2, Kneba M.1, Brüggemann M.1 2. Medizinische Klinik, Universitätsklinikum Schleswig-Holstein, Hämatologie Labor, Kiel, Germany, 2Medizinische Klinik II, Universitätsklinikum, Frankfurt am Main, Germany 1

Introduction: Minimal residual disease (MRD) has been proven to be the most powerful prognostic factor in ALL. The current molecular gold standard for MRD quantification is the allele specific real-time quantitative PCR (ASO RQ-PCR) of clonal immunoglobuline (IG) and T-cell receptor (TR) gene rearrangements. However, quantification is mostly limited to 1xE-04 and PCR signals outside the quantitative range (QR) are regarded as less specific and neither judged as complete MRD-response nor -persistence. Next-generation sequencing (NGS) might overcome this limitation and potentially allows a quantification of low-level MRD. We have thus performed a head-to-head comparison of ASO RQ-PCR and NGS on patients within the German Multicenter ALL Trial (GMALL). Methods: 128 samples (58 diagnostic and 70 follow-up) of 70 patients were analyzed using the Illumina MiSeq-platform. Using universal primer sets IGH V-, D- and J-gene segments were amplified. Amplicons were sequenced and analyzed using standardized algorithms for clonotype determination. Leukemia specific clonotypes were identified based on their frequency in the diagnostic sample and quantified in positive, not quantifiable RQ-PCR samples at therapy week 16 using internal reference DNA as a measure of clone level among all leukocytes. Results: A clonal rearrangement suitable for further MRD-analysis was identified in 57/58 diagnostic samples. In each sample the rearrangement used for RQ-PCR-diagnostics was re-identified by NGS, with 95% of the NDN-sequences matching. The Sanger-sequenced routine marker represented the major NGS clone in 52 cases, it correlated to a minor NGS clone in 6 cases, where an additional more dominant clonal rearrangement could be identified. In 25/70 follow up-samples NGS detected MRD, whereas in 42/70 samples no leukemia-specific read was identified, thus leading to NGS-MRD-negativity. 3 samples showed specific reads, but below the prior defined sensitivity level, and were classified as positive< QR. Conclusion: Our results indicate that NGS is a promising tool to screen diagnostic samples for clonal IGH markers in ALL patients, with a high concordance to routine Sanger-sequencing. In follow-up samples with questionable low RQ-PCR positivity NGS specifically re-identified the leukemia specific clone in 36% of cases. Whether discrepant NGS-/RQPCR+ results are attributable to false RQ-PCR positivity or reflect a higher sensitivity of RQ-PCR has to be clarified by clinical correlation. Disclosure: No conflict of interest disclosed.

Abstracts

V685

Impact of gender on outcome after chemoimmunotherapy with fludarabine, cyclophosphamide and rituximab (FCR) or bendamustin plus R (BR) in CLL patients: a pooled analysis of the German CLL Study Group (GCLLSG) Al-Sawaf O.1, Kluth S.1, Bahlo J.1, Hopfinger G.2, Fink A.M.1, Cramer P.1, Maurer C.1, Bergmann M.3, Dreyling M.4, Lange E.5, Kneba M.6, Stilgenbauer S.7, Kiehl M.G.8, Jäger U.9, Wendtner C.M.3, Fischer K.1, Hallek M.1,10, Eichhorst B.1 Uniklinik Köln, Centrum für Integrierte Onkologie Köln Bonn, Klinik 1 für Innere Medizin, Deutsche CLL Studiengruppe, Köln, Germany, 2Medizinische Universität Wien, Universitätsklinik für Innere Medizin 1, Wien, Austria, 3 Klinikum München-Schwabing, Klinik für Hämatologie, Onkologie, Immunologie, Palliativmedizin, Infektiologie und Tropenmedizin, München, Germany, 4Klinikum der Universität München / Medizinische Klinik und Poliklinik III, München, Germany, 5Evangelisches Krankenhaus Hamm, Klinik für Hämatologie, Onkologie und Palliativmedizin, Hamm, Germany, 6 Universitätsklinikum Schleswig-Holstein, Klinik für Hämatologie, Onkologie und Palliativmedizin, Kiel, Germany, 7Uniklinik Ulm, Klinik für Innere Medizin III, Ulm, Germany, 8Klinikum Frankfurt (Oder), Medizinische Klinik 1, Frankfurt (Oder), Germany, 9Allgemeines Krankenhaus Wien, Klinische Abteilung für Hämatologie und Hämostaseologie, Wien, Austria, 10Uniklinik Köln, CECAD Cluster of Excellence Cellular Stress Responses in Aging-Associated Diseases, Köln, Germany 1

Introduction: Chemoimmunotherapy with FCR is the current standard for fit CLL patients without unfavourable TP53 deletion/mutation. Due to better tolerability BR might be used in pts aged 65 or older though it is less effective. In CLL women have a better overall survival (OS) than male pts. The aim of this study was to systematically evaluate the impact of gender on the progression-free (PFS) and OS after therapy with FCR or BR. Methods: Data from three clinical trials (phase II and III) of the GCLLSG including 1078 pts were analysed. 683 pts received frontline therapy with FCR (404 within CLL8 (FCR vs FC), 279 within CLL10 (FCR versus BR)) and 395 received BR (278 within CLL10 and 117 within CLL2M). Laboratory markers, genetic parameters as well as event related data were pooled and compared with regard to gender. Kaplan-Meier survival curves were calculated and compared by non-stratified log-rank test. Hazard ratios (HR) and 95% confidence intervals (CI) were calculated using Cox regression modelling. Results: 683 pts received FCR and 395 pts received BR. Median age was 61 years (30–81). 291 (27%) pts were female, 787 (73%) were male. The distribution of Binet stages was similar between women and men, as well as, frequency of unfavourable mutations (17p deletion, 11q deletion, unmutated IGHV) or elevated serum parameters (thymidine kinase, ß2 microglobuline) except of trisomy 12 (18.7% in female vs 9.8% in male). Frequency of dose reductions and median number of treatment cycles did not differ between genders. 5-year- with BR was 89.7% in women vs 57.6% in men (HR:1.99, 95%CI = 0.96–4.13; p = 0.06) and 81.9% vs 79.2% with FCR (HR 1.26, 95%CI = 0.86–1.84; p = 0.24). 5-year-PFS was significantly longer in women after FCR (55.8% vs 44.5%; HR:1.28, 95%CI = 1.00–1.646; p =0.050) as well as after BR (29.0% vs 23.3%; HR:1.46, 95%CI = 1.043– 2.052; p = 0.028). While BR was significantly inferior to FCR in male pts (5 years PFS 23.3% vs. 44.5%; HR 1.658, CI 1.338–2.056; p < 0.001) the difference was statistically not significant in females (5 years PFS 29.0% vs. 55.8%; HR 1.381,CI 0.941–2.026; p = 0.099). Conclusion: According to this pooled analysis, women had a longer PFS after chemoimmunotherapy either with FCR or BR confirming the better prognosis of female CLL pts also regard to efficacy of treatment. Additional data on the frequency and type of toxicities in male and female pts will be presented at the meeting.

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Clonal hierarchies and dynamic evolution in Myelodysplastic Syndromes (MDS) upon therapy Mossner M.1, Jann J.C.1, Wittig J.1, Nolte F.2, Fey S.1, Nowak V.1, Obländer J.1, Pressler J.1, Palme I.1, Xanthopoulos C.1, Boch T.1, Metzgeroth G.1, Röhl H.3, Witt S.H.4, Dukal H.4, Klein C.5, Schmitt S.6, Gelß P.7, Platzbecker U.8, Bulycheva E.8, Fabarius A.1, Blum H.9, Schulze T.J.10, Meggendorfer M.11, Haferlach C.11, Trumpp A.5,12, Hofmann W.K.1, Medyouf H.13, Nowak D.1 Medizinische Fakultät Mannheim der Universität Heidelberg. III. Medizinische Klinik, Hämatologie und Onkologie, Mannheim, Germany, 2St. Hedwig Krankenhaus, Berlin, Germany, 3Orthopädische und Unfallchirurgische Klinik, Universitätsmedizin Mannheim, Mannheim, Germany, 4Zentralinstitut (ZI) für Seelische Gesundheit, Mannheim, Germany, 5Abteilung für Stammzellen und Krebs, Deutsches Krebsforschungszentrum (DKFZ), Heidelberg, Germany, 6 Zytometrie Core Facility, Deutsches Krebsforschungszentrum (DKFZ), Heidelberg, Germany, 7Department of Mathematics and Computer Science, Freie Universität Berlin, Berlin, Germany, 8Klinik für Hämatologie und Onkologie, Universitätsklinik Carl Gustav Carus, Dresden, Germany, 9Laboratory for Functional Genome Analysis, Gene Center of the LMU, München, Germany, 10 Institut für Transfusionsmedizin und Imunologie, Medizinische Fakultät Mannheim der Universität Heidelberg, Mannheim, Germany, 11Müncher Leukämielabor (MLL), München, Germany, 12HiStem GmbH, Heidelberg, Germany, 13Georg-Speyer-Haus, Frankfurt a.M., Germany 1

Fig. 1. Disclosure: Othman Al-Sawaf: Other Financial Relationships: Travel grant from Gilead. Barbara Eichhorst: Advisory Role: Janssen, Gilead, AbbVie, GSK, Celgene, Roche, and Mundipharma; Other Financial Relationships: Janssen, Gilead, AbbVie, GSK, Celgene, Roche, and Mundipharma

Introduction: Clonal evolution is believed to be a main driver for progression of cancer and implicated in facilitating resistance to drugs. In this context the hierarchical organization of malignant clones in the hematopoiesis of myelodysplastic syndromes (MDS) and its impact on response to drug therapy remain poorly understood. Methods: Using next generation sequencing (NGS) of patient and xenografted cells we evaluated the intratumoral heterogeneity in n = 47 patients and reconstructed mutational trajectories in n = 39 cases suffering from MDS (n = 37) and CMML-1 (n = 2). Genomic DNA was isolated from patient bone marrow (BM) / CD34+ cells and patient-matched BM-derived mesenchymal stromal cells (MSCs) as non-hematopoietic germline controls. Matched DNA pairs from at least 2 time points per patient were subjected to whole exome sequencing (WES) with an average coverage of 100x on an Illumina HiSeq 2000 sequencing device. In addition, a custom amplicon-based MiSeq panel was applied to BM samples for deep sequencing of 24 commonly mutated genes in MDS. For validation of mutational events we performed targeted quantitative pyrosequencing for point and small INDEL mutations and capillary fragment analysis for longer INDEL variants. Cytogenetic lesions were quantified using interphase-FISH and a newly developed multiplex microsatellite assay for measurement of loss of heterozygosity. Results: We identified linear but also branching evolution paths and confirmed on a patient-specific level that somatic mutations in epigenetic regulators and RNA splicing genes frequently constitute isolated disease-initiating events. Using high-throughput exome- and deep sequencing we analyzed 77 chronologically acquired samples from 13 patients covering a cumulative observation time of 66 years of MDS disease progression. Our data revealed highly dynamic shaping of complex oligoclonal architectures specifically upon treatment with lenalidomide and other drugs. Despite initial clinical response to treatment, patients’ marrow persistently remained clonal with rapid outgrowth of founder-, sub- or even fully independent clones indicating an increased dynamic rate of clonal turnover. The emergence and disappearance of specific clones frequently correlated with changes of clinical parameters highlighting their distinct and far-reaching functional properties. Conclusion: Our data substantiate a need for regular broad molecular monitoring to guide clinical treatment decisions in MDS. Disclosure: No conflict of interest disclosed.

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Cytotoxic activity and molecular mechanisms of action of the marine triterpene glycoside frondoside A in germ cell tumors Alsdorf W.H.1, Dyshlovoy S.1, Otte K.1, Hausschild J.1, Bokemeyer C.1, Honecker F.2, von Amsberg G.1 II. Medizinische Klinik Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany, 2ZeTuP St. Gallen, St. Gallen, Switzerland 1

Introduction: Metastatic germ cell tumors (GCT) are curable by cisplatin (Cis) based combination chemotherapy. However, a subset of patients develops Cis resistant disease resulting in a very poor prognosis. Frondoside A (FrA) is a marine triterpene glycoside molecule derived from the sea cucumber Cucumaria frondosa. It has shown substantial anti-proliferative activity in several malignancies in preclinical experiments. Methods: Cytotoxic activity of FrA was measured by MTT and colony formation assays. In vitro combination treatment (FrA+Cis) was analyzed by the Chou-Talalay method. Apoptosis and Cell cycle analysis were performed by FACS propidium iodide (PI) and Annexin V/FITC PI staining. Apoptosis and autophagy signaling was investigated by western blotting. Cellular subfractionation was done to examine nuclear translocation of apoptosis inducing factor (AIF). Results: In vitro treatment of GCT cell lines NCCIT and 2102EP with FrA exhibited IC50 values of 0.5 µM. Importantly, Cis resistant sublines NCCIT-R and 2102EP-R were equally sensitive to FrA (IC50 0.4–0.5 µM) Colony formation was significantly inhibited by FrA at 0.125 µM. Combination therapy with FrA and Cis led to an additive cytotoxic effect. Contrary to Cis, FrA induces caspase-independent apoptosis. Thus, cleaved Caspase 3 and cleaved PARP proteins were only upregulated after exposure to Cis but not FrA. FACS analysis of GCT cells after pretreatment with the caspase inhibitor z-VAD(OMe)-fmk confirmed this finding as tumor cells treated with FrA still exhibited extensive apoptosis despite caspase inhibition. Unlike Cis, FrA did not induce DNA damage as measured by induction of phospho-H2AX formation. To further examine the mechanism of caspase-independent apoptosis, cellular subfractions of GCT cells were investigated for accumulation of apoptosis-inducing factor (AIF), a mediator of caspase-independent apoptosis. FrA treatment led to increase of AIF within the nuclear fraction. In addition, FrA was identified as an inhibitor of pro-survival autophagy demonstrated by increase of LC3B-II and SQSTM/p62 levels. Conclusions: FrA has distinct molecular effector mechanisms as compared to Cis and displays substantial anti-tumor efficacy in vitro against Cis resistant GCT cells. FrA was identified as inhibitor of pro-survival autophagy and induces caspase-independent apoptosis by activation of AIF. These findings highlight autophagy and AIF pathways as novel therapeutic targets in GCT. Disclosure: No conflict of interest disclosed.

organogenesis and proper function of the hematopoietic stem cell niche. However, CXCR4 is also involved in survival, proliferation and dissemination of cancer cells and its expression is associated with adverse prognosis in a broad range of malignancies, including acute leukemia (AML, ALL). Acute leukemias are often difficult to treat and highly active therapies are needed in particular in the relapsed/refractory setting to effectively reduce disease burden before allogeneic stem cell transplantation (alloSCT). Methods: We used patient-derived xenograft (PDX) mouse models of acute leukemia to evaluate the efficacy and toxicity of a CXCR4-targeted peptide receptor radiotherapy (PRRT) theranostic approach with the CXCR4 PET tracer 68Ga-Pentixafor and its b-emitting therapeutic counterpart 177Lu-Pentixather. Results: 68Ga-Pentixafor enabled visualization of CXCR4 positive leukemic burden in spleen and bone marrow of acute leukemia PDX by PET imaging. In T-ALL PDX, CXCR4-directed PRRT with 177Lu-Pentixather rapidly and effectively distributed to leukemia harboring organs. 177 Lu-Pentixather therapy resulted in efficient reduction or eradication of leukemic infiltration in peripheral blood, spleen and bone marrow. Spleen size was reduced dramatically as early as 24h after initiation of PRRT. Treated mice suffered severe suppression of bone marrow function as evidenced by therapy induced severe cytopenia affecting mature CD45+ blood cells and colony-forming unit potential of progenitors in the bone marrow. Our data indicate that CXCR4 targeting and the inevitable and desired cross-fire effect of 177Lu-Pentixather PRRT alone may be sufficient to induce myeloablation and could serve as a valuable addition to conditioning protocols in alloSCT. Evaluation of this concept within host conditioning in alloSCT is under investigation. Conclusion: In conclusion, our work provides first evidence for the efficacy of the novel CXCR4-directed agent 177Lu-Pentixather in an acute leukemia PDX model. Importantly, these findings can directly be translated into clinical studies in patients and will help to design phase 1 protocols. Disclosure: No conflict of interest disclosed.

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Metastasiertes Mammakarzinom. Gemeinsames Symposium mit der Deutschen Gesellschaft für Senologie V693

Ribociclib + letrozole for postmenopausal women with hormone receptor-positive (HR+), HER2-negative, advanced breast cancer (ABC) who received no prior therapy for advanced disease Janni W.1, Nusch A.2, Grischke E.-M.3, Marschner N.4, Wilke J.5, Decker T.6, Abenhardt W.7, Kurbacher C.8, Overkamp F.9, Just M.10, Kümmel S.11, Marmé F.12,13, Xuan F.14, Miller M.14, Sonke G.S.15 Universitätsklinikum Ulm, Department of Obstetrics and Gynecology, Ulm, Germany, 2Medical Center for Hematology and Internal Oncology, Velbert, Germany, 3Universitätsklinikum Tübingen, Department of Obstetrics and Gynecology, Tübingen, Germany, 4Medical Center for Interdisciplinary Oncology and Hematology, Freiburg i.Br., Germany, 5Medical Center for Oncology, Fürth, Germany, 6Medical Center for Oncology, Ravensburg, Germany, 7Münchner Onkologische Praxis im Elisenhof,MVZ, München, Germany, 8Department of Gynecological Oncology, Medical Center Bonn Friedensplatz, Bonn, Germany, 9 Oncologianova GmbH, Recklinghausen, Germany, 10Medical Center for Oncology, Bielefeld, Germany, 11Breast Unit, Kliniken Essen-Mitte, Essen, Germany, 12National Centre for Tumour Diseases, Heidelberg, Germany, 13 Universitätsklinikum Heidelberg, Department of Gynecologic Oncology, Heidelberg, Germany, 14Novartis Pharmaceuticals Corporation, East Hanover, United States, 15The Netherlands Cancer Institute, Amsterdam, Germany 1

V688

CXCR4-directed peptide receptor radiotherapy with 177LuPentixather effectively clears leukemia in patient-derived xenografts Habringer S.1,2, Herhaus P.1, Schottelius M.3, Istvanffy R.1, Götze K.1,2, Oostendorp R.1, Grigoleit G.U.4, Lapa C.5, Jeremias I.6, Schwaiger M.7, Peschel C.1,2, Wester H.-J.3, Keller U.1,2 3. Medizinische Klinik, Klinikum rechts der Isar der TUM, München, Germany, DKFZ Heidelberg, DKTK, München, Germany, 3Pharmazeutische Radiochemie, Technische Universität München, München, Germany, 4Medizinische Klinik und Poliklinik II, Universitätsklinikum Würzburg, Würzburg, Germany, 5 Klinik und Poliklinik für Nuklearmedizin, Universitätsklinikum Würzburg, Würzburg, Germany, 6Helmholtz Zentrum München, München, Germany, 7 Nuklearmedizinische Klinik, Klinikum rechts der Isar, Technische Universität München, München, Germany 1 2

Introduction: CXC-motif Chemokine Receptor 4 (CXCR4) is a transmembrane receptor with pivotal roles in physiologic processes like cell migration,

Abstracts

Introduction: Cyclin-dependent kinase (CDK) 4/6 inhibition can block cell proliferation and growth in endocrine-resistant cells and may help overcome ET resistance. Ribociclib, an orally bioavailable, selective, small-molecule inhibitor of CDK4/6, has exhibited antitumor activity as part of doublet or triplet therapy (ribociclib + ET ± PI3K inhibitor or everolimus) in estrogen receptor-positive breast cancer models, and in the

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clinical setting (Munster et al ASCO Breast 2014: 143; Bardia et al EORTC 2014: P072; Juric et al SABCS 2015: P3-14-01). The phase 3, randomized, double-blind, international, MONALEESA-2 study is evaluating the efficacy and safety of ribociclib + letrozole vs placebo + letrozole in patients with HR+, HER2-negative ABC who have received no prior systemic therapy for advanced disease. Methods: Postmenopausal women (n = 668) with confirmed HR+, HER2-negative ABC with measurable disease or ≥1 predominantly lytic bone lesion, ECOG performance status ≤1, and adequate bone marrow and organ function were enrolled. No prior systemic therapy for ABC or prior CDK4/6 inhibitor treatment were allowed; (neo)adjuvant therapy was allowed if disease free interval was >12 months after non-steroidal aromatase inhibitor treatment. Patients were randomized 1:1 to receive ribociclib (600 mg/day on Days 1–21 of each 28-day cycle) + letrozole (2.5 mg/day continuously) or placebo + letrozole, stratified by the presence of liver and/or lung metastases. Primary endpoint is progression-free survival (PFS) by local radiological assessment. The key secondary endpoint is overall survival; other secondary endpoints include overall response rate, clinical benefit rate, and safety. Expected results: Independent Data Monitoring Committee recommended stopping the trial early as it met the primary endpoint, significantly extending PFS compared to letrozole alone, at the pre-planned interim analysis. Data from the interim efficacy analysis, including PFS and supportive secondary endpoints, will be presented. Expected conclusions: Ribociclib + letrozole significantly improved PFS vs letrozole monotherapy in postmenopausal women with HR+, HER2-negative ABC who had received no prior therapy for advanced disease. Disclosure: Wolfgang Janni: Financing of Scientific Research: Ja, Honorare für Teilnahme an Novartis Steering Committee Pipeline Mammakarzinom und Honorare für Vorträge in Satellitensymposien; Expert Testimony: Ja, teilnehmendes Zentrum an MONALEESA-2, Unterstützung Detect-Studien Gabe Sonke: No conflict of interest disclosed.

Wissenschaftliches Symposium

Allogene Stammzelltransplantation: aktuelle Konzepte V694

Tailoring conditioning intensity according to comorbidities and risk of relapse Bornhäuser M.1, Stölzel F.1, Schetelig J.1 Med. Klinik und Poliklinik I, Dresden, Germany

The past two decades have witnessed the development of various conditioning therapies whch can be successfully applied before allogeneic hematopoietic cell transplantation (HCT) in older patients and those with reduced organ function. These regimens typically combine Purine-analogues (e.g. fludarabine) with classical alkylators (busulfan, melphalan etc.). Minimal conditioning therapies with 2 Gy total-Body Irradiation and fludarabine and/or cyclophosphamide seem to be attractive for patients with non-malignant diseases. As patients with AML and MDS face a considerable risk of relapse even in the context of allogeneic HCT, intensive conditioning regimens with tolerable toxicity seem to be preferable. Recent randomized trials suggest that patients with AML and MDS are at an increased risk of treatment failure if the intensity of the preparative regimen is below a certain threshold. On the other Hand, comparisons of classical double alkylator regimens with Purine analogue-based but still intensive regimens in HLA-matched settings suggest a significant reduction in non-hematologic toxicity and non-relapse mortality with comparable long-term outcomes after RIC. Furthermore, strategies combining salvage chemotherapy with early transplantation during aplasia (e.g FLAMSA-RIC; 3+7 +RIC etc.) have been established for patients with high-risk disease. Future Trials will have to identify the ideal conidtioning intensity used before haploidentical transplantation where high-dose cyclophosphamide is applied post-Transplantation.

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In summary, patients up to the Age of 65 who are likely to tolerate classical conditioning intensity and have a considerable risk of disease relapse should receive a more initensive regimen. With the advent of optimized supportive care, the use of intensive but moderately toxic regimens can be envisioned even in patients > 65. Disclosure: Martin Bornhäuser: Advisory Role: Jazz Pharamaceuticals, Alexion; Financing of Scientific Research: Celgene, GWT Dresden, Novartis, Jazz, Alexion, MSD, Cellex GmbH Johannes Schetelig: Employment or Leadership Position: DKMS

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Mikroskopierkurs I V697

WHO classification 2016 for the myelodysplastic syndromes (MDS): how to proceed Germing U.1, Strupp C.1 Heinrich-Heine University Düsseldorf, Department of Hematology, Oncology and Clinical Immunology, Düsseldorf, Germany 1

A refinement of the classification of the MDS has been proposed by the WHO working group in 2016 including new names of the categories, the reintroduction of a category multilineage dysplasia with ring sideroblasts, the introduction of SF3B1 mutations as a classifier for patients with < 15, but >5% ring sideroblasts, the inclusion of MDS with del(5q) and one additional non chromosome 7 anomaly in to the MDS del(5q) group, and the definition to assess the medullary blast count, namely the blasts should be based on all nucleated cells neglecting the amount of erythroid cells. Besides that, cell counts influencing the categorization have been introduced. As a result of the new WHO proposals, haematologists, pathologists, human geneticists, and laboratorists should be taken into account the following procedures for a proper diagnosis of MDS: 1. cell counts (number of cytopenias, MDS-U or other MDS type?) 2. at least 2 full differential counts (monocytosis, CMML 0, 1, 2 ?, percentage of peripheral blasts, MDS-U, MDSEB 1,2 ?) 3. assessment of the number of dysplastic lineages by single cell morphology (>10% dysplastic cells per lineage required, assessment of presence and percentage of ring sideroblasts (iron staining) (MDSSLD or MDSMLD, MDSSLD-RS or MDSMLD-RS?) 4. assessment of the percentage of medullary blasts based on all nucleated cells in the marrow (exact percentage needed for IPSS-R, 0–2, 3–4, 5–9, 10–19%) 5. analysis of SF3B1 at least in cases with some, but less than 15% ring sideroblasts 6. karyotype banding of at least 20, better 25 metaphases (MDS del(5q)?, aberration in case of weak signs of dysplasia, MDS-U?) After producing or collecting these parameters, a categorization of the MDS cases according to the WHO 2016 proposals is possible, providing important information with regard to the biology of the disease, prognostic behavior, and therapeutic intervention. Disclosure: No conflict of interest disclosed. V698

WHO classification 2016 for AML Haferlach T.1 MLL Munich Leukemia Laboratory, München, Germany

In April 2016 Arber et al. (Blood, prepublished online April 11, 2016) published the revision of the WHO classification of myeloid neoplasms and acute leukemia. This is an important step forward to better describe and use newest clinical but especially cytogenetic and molecular genetic information. We first have to separate patients with “therapy-related myeloid neoplasms” following chemotherapy and / or radiotherapy. The next step includes patients with “AML and recurrent genetic abnormalities”, defined

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by cytogenetic translocations in most cases and also by the respective involved genes. In addition, NPM1 mutated AML and CEBPA double mutated AML are now full entities. Two new provisional entities have been implemented: AML with BCR-ABL1 and AML with mutated RUNX1. The second category defines patients with “AML and myelodysplasia-related changes”. Those cases are defined by specific cytogenetic abnormalities, in many cases by complex karyotypes defined as 3 or more abnormalities. More than 10 other specific cytogenetic aberrations are listed here. Further, those patients can be classified into the category of “AML with myelodysplasia-related changes” if 2 or 3 cell lines demonstrate dysplasia (50% or more of the cells show dysplastic features). Of note, cases with NPM1 mutations or CEBPA double mutations showing dysplasia in 2 or 3 cell lines (multi lineage dysplasia, MLD) will still be categorized according to their molecular aberration and not in the group of AML with myelodysplasia-related changes. All the rest AML cases (around 15%, depending on age of the cohort) will then be classified according to their morphological appearance only, following somehow the old FAB classification. A new classification has been defined for the former AML M6 group, now pure erythroid leukemia. The erythropoiesis still has to be 50% or more of the bone marrow nucleated cells but the myeloid blasts will be counted in an absolute count and not relative to all the rest of the non-erythroid cells anymore. Finally, the WHO 2016 defines a new group of “myeloid neoplasms with germline predisposition”, including patients with germline CEBPA mutation or germline DDX41 mutation or germline ANKRD26 mutation or germline ETV6 mutation or germline GATA2 mutation. The inclusion of the category “with germline predisposition” is a very important step forward for our understanding of AML and makes it necessary that clinicians much more than before investigate the patients history. Disclosure: Torsten Haferlach: Employment or Leadership Position: MLL Muich Leukemia Laboratory

Wissenschaftliches Symposium

Survivorship, Rehabilitation – psychosoziale Belastungen V700

Psychological and social side effects in young or adolescent cancer patients Seifart U.1 Klinik Sonnenblick, Marburg, Germany

In Germany 18000 pts fall ill at the age of 18-39 years, per year with cancer. From these approximately 80%, on account of improved oncological therapy, survive their cancer illness. For this “longterm survivor” long time side effects caused by therapy pose a considerable problem, or a considerable restriction of quality of life. These enclose beside the known symptoms like Fatigue, chemotherapy induced polyneuropathy or cardiotoxicity, the question of infertility for patients who does not fixed their family planning as a considerable problem. Beside the psychological effects the patients may face financial problems also. Thus cryopreservation is still not financed by health insurance. On account of not cleared mechanisms disturbances of concentration and retention may occur. A frightening fact for many young patients. In addition, these aspects, as well as the changed body picture can complicate the integration in the circle of acquaintances or peer group considerably what can lead to psychic load situations again. However, these somatic and psychological side effects of oncological therapies cause, on account of the achievement decrease, financial and social restrictions due to difficulties in return to work. Young adults may have the additional problem that they have not concluded their professional training or study. The menace of the financial existence of the patient and sometimes also of the family, cause a considerable load of quality of life and shows an often underestimated problem for young cancer patients. Disclosure: No conflict of interest disclosed.

Abstracts

V701

Occupational perspective and return to work in cancer survivors Rick O.1 Klinik Reinhardshöhe, Onkologische Rehabilitation, Bad Wildungen, Germany

Unfavorable cancer and treatment consequences include psychological distress and different physical and functional disabilities that may adversely affect work ability and employment status. Compared to healthy controls previous studies suggested that cancer increases the risk of unemployment among survivors. A combined assessment of reports from various countries reveals that 63% (range: 24% to 94%) of cancer survivors who are of working age return to work after being unable to work for an average of five month. Between 26% and 53% of patients lost their job or gave up work in the first 6 years after diagnosis. Another meta-analysis of 36 controlled studies showed that 34% of the long term survivors were unemployed compared with 15% of healthy controls, corresponding to a relative risk of 1.37 (95% CI 1.21 to 1.55). Therefore, many survivors changed jobs, worked a reduced number of hours per day, and experienced a drop in pay. Furthermore, in Norway 31% of the cancer patients work with reduced physical and 23% with reduced mental function. The percentages of patients who return to work varies considerable in the different social systems. Whereas in Germany nearly 60% of patients with breast cancer are employable, around 73% of the patients in the USA are back at work. The most likely reason for this fact is that the German social system makes it possible to make a special allowance for the health sequelae of cancer, with up to 78 weeks sick pay from the health insurance, and a partial or complete disability pension. In contrast, after medical rehabilitation 76% of the cancer patients in Germany return to work. Unfavorable prognostic factors for return to work are tumor type, cancer therapy, physical and mental impairments, sociodemographic and work-related conditions. The immediate conditions in the workplace and the support provided by the employer and the social framework are crucial promoting factors for return to work. A pooled study of 15 randomized clinical trials with 1835 cancer patients showed that multidisciplinary interventions involving physical, psycho-educational and vocational components led to higher return to work rates than usual care. In conclusion, the awareness of health professionals, employer and the social framework regarding barriers of return to work may improve the early identification of cancer survivors at risk of prolonged time to work ability and may allow early supportive interventions. Disclosure: No conflict of interest disclosed.

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Therapieoptimierung der akuten lymphatischen Leukämie V709

Mutation analysis and minimal residual disease detection within Philadelphia-Chromosome positive (Ph+) acute lymphoblastic leukemia – clinical relevance Pfeifer H.1 Universitätsklinikum Frankfurt, Goethe-Universität, Medizinische Klinik II, Abt. für Hämatologie/Onkologie, Frankfurt, Germany 1

Tyrosine kinase inhibitors (TKI) in combination with chemotherapy are the gold standard for front-line therapy of Ph(+) ALL, with remission rates exceeding 90%. These treatment regimens led to a better outcome compared with historic controls, but most patients who do not undergo allogeneic stem cell transplantation (SCT) relapse. Several studies have demonstrated that minimal residual disease (MRD) assessment in childhood and adult acute lymphoblastic leukemia (ALL) significantly correlates with clinical outcome. Whereas molecular monitoring of minimal residual disease levels is prognostic relevant for patients with Ph-negative ALL and should be used to guide treatment, the role of MRD assessment

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in Ph(+) ALL is still unclear. MRD detection is particularly useful for evaluation of early treatment response, but also to monitor disease before and after stem cell transplantation, for early assessment of an impending relapse and in the setting of salvage treatment. E.g. within the GMALL trials, intermediate and low MRD positivity pre SCT showed no prognostic impact on Overall Survival and Disease Free Survival. After SCT regular MRD measurement may predict a relapse and is already used to guide therapy including change of tyrosine kinase inhibitor. In addition, international standardization and quality control efforts are ongoing to ensure comparability of results. If a relapse occurs – acquired resistance on TKI treatment is often associated with mutations in the bcr-abl tyrosine kinase domain. This may be detected at low frequency during/prior to TKI treatment in a subset of patients. Mutation analysis during treatment relies increasingly on highly sensitive PCR techniques and may assist in treatment decisions, eg. in case of molecular relapse. Second and third generation TKIs, show activity against most of the bcr-abl tyrosine kinase domain mutations involved in acquired imatinib resistance, but clinical benefit is generally short-lived. Therefore, still SCT in first complete remission (CR) is considered to be the best curative option. Disclosure: Heike Pfeifer: Advisory Role: Ariad; Financing of Scientific Research: Novartis, BMS, Amgen; Expert Testimony: Novartis, BMS; Other Financial Relationships: Reisekostenerstattung von Novartis, Amgen V710

Innovative approaches to stem cell transplantation Handgretinger R.1 Universitätskinderklinik, Hämatologie/Onkologie, Tübingen, Germany

In contrast to T-cell replete allogeneic transplantation, the ex vivo-depletion of T-cells from the graft allows the transplantation of HLA-identical and haploidentical stem cells with minimal or with the complete omission of posttransplant immunosuppression for prevention of Graft-versusHost Disease (GvHD). While CD34+ positive selection was used initially for indirect depletion of T-lymphocytes, the negative depletion of T-cells from mobilised peripheral stem cells (PBSCs) is increasingly used. Especially the depletion of alpha beta T-cell receptor (TcRαβ) positive T-cells and/or the depletion of CD45RA positive naive T-cells has been shown to be associated with a faster immune reconstitution compared to transplantation of purified CD34+ stem cells. In clinical studies of haploidentical transplantation of TcRαβ depleted stem cells, a low incidence of GvHD and a low rate of transplant-related mortality (TRM) was seen. With negatively depleted grafts, high numbers of Natural Killer (NK) cells and γδ+ T-lymphocytes are cotransplanted together with the stem cells, which leads to a rapid posttransplant expansion of NK cells and γδ+ T-cells. These cells are not alloreactive and might protect the patients in the early posttransplant phase from severe infections. There is evidence that NK cells as well as γδ+ T-cells have anti-leukemic effects and might also protect the patients from relapse. The rapid expansion of NK- and γδ+ cells facilitate further posttransplant strategies to prevent or treat severe viral infections with donor-derived virus-specific T-cells and promising clinical responses have been seen with virus-specific T-cells. Since a major cause for therapy failure is relapse of the underlying disease, anti-leukemic strategies in the early posttransplant phase can be employed. Such strategies include the use of zoledronic acid to activate γδ T-cells, the use of monoclonal antibodies to stimulate the antibody-dependent cellular cytotoxicity (ADCC) of NK cells and γδ+ T-cells or the application of bispecific T-cell engagers (BiTEs). In additon, cellular therapeutic strategies include the adoptive transfer of donor-derived NK cells, tumor-specific T-cells or in the future the use of donor-derived chimeric antigen receptor (CAR) T-cells. In summary, allogeneic transplantation of ex vivo T-cell depleted grafts is a platform for further posttransplant humoral and cellular therapy strategies after HLA-identical and haploidentical transplantation.

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Disclosure: Rupert Handgretinger: Advisory Role: ALL Steering Comittee, Amgen.; Honoraria: Patentanteil an der TcRalpha/beta Depletionsmethode; Financing of Scientific Research: Honorare für Vorträge von Amgen, Miltenyibiotec; Other Financial Relationships: Reisekostenerstattung von Amgen, Miltenyibiotec

Wissenschaftliches Symposium Liquid Biopsy V713

Chemosensitivity testing: a critical perspective and review Zenz T.1 Molekulare Therapie in der Hämatologie und Onkologie / NCT / DKFZ / Uniklinik Heidelberg, Heidelberg, Germany 1

Response to anti-cancer agents is often restricted to subsets of patients, but the recognition of factors underlying this heterogeneity and the identification of biomarkers is incomplete. There is a need for platforms that can comprehensively map drug responses, identify associated biomarkers and provide hypotheses for mechanisms underlying variable response. Determinants of drug response have been mapped in immortalized cancer cell lines (http://www.cancerRxgene.org; http://www.broadinstitute. org/ccle; http://www.broadinstitute.org/ctrp), and recent technology improvements allow increased throughput. However, suitable cell lines are not available for all cancers, including chronic lymphocytic leukaemia (CLL), the most common adult leukaemia in Western countries. Shortterm cultures of primary cells have advantages over immortalized cells, as clonal selection can be avoided. Cohorts of such patient-derived samples represent the natural genetic and phenotypic diversity of cancers, including rarer mutations (the “long tail”) and combinatorial patterns of mutations. Responses of primary tumour cells to panels of inhibitors ex vivo have recently been used to derive individualized therapeutic options for the donating patients. 20 years after an initial surge of chemosensitivity testing, there is now growing evidence that drug response phenotypes probed ex vivo can recapitulate clinical biomarkers, identify novel disease-specific sensitivities and provide the comprehensive maps linking drug response phenotypes to molecular features e.g. of leukemia. The presentation will review the current role for drug sensitivity testing for a novel generation of precision medicine tools. Disclosure: No conflict of interest disclosed.

Freier Vortrag

B-Zell-Lymphome, experimentell V715

Optimizing 25-OH-Vitamin D3 serum levels for Rituximaband Obinutuzumab-mediated antibody-dependent cellular cytotoxicity Acker F.1, Bittenbring J.T.1, Pfreundschuh M.1, Neumann F.1 Universitätskliniken des Saarlandes, Innere Medizin, Homburg/Saar, Germany

Vitamin D3 deficiency impairs the rituximab-mediated antibody-dependent cellular cytotoxicity (ADCC) and the outcome of elderly patients with diffuse large B-cell lymphoma (Bittenbring et al., J Clin Oncol. 2014 Oct 10; 32(29):3242-8) treated with R-CHOP. The aim of this study was to determine the optimal 25-OH-vitamin D3 (VD3) level for rituximab- and obinutuzumab-mediated ADCC. Ten individuals (5 males, 5 females; mean age: 67.7 years, range 41-79) without malignant disease or immunosuppression were included in this study after written informed consent. PBMC were isolated and CD16+ NK cells were separated by depleting all non-NK-cells magnetically. ADCC activity of the NK cells was tested against CD20-expressing Daudi cells without or after anti-CD20-antibody treatment with rituximab and obi-

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nutuzumab, respectively. Cytotoxic activity was assessed by LDH release by the target cells. NK cells were studied at four different VD3 serum levels: : < 20 ng/ml; 30 ng/ml, 65 ng/ml, and 90 ng/ml). To achieve these levels the probands were substituted with cholecalciferol. The median VD3 serum level before substitution was 10 ng/ml. The only formula for achieving predefined VD3 serum levels published by van Groningen et al. (Eur J Endocrinol. 2010; 162:805-11) failed to achieve the predetermined VD3 levels. The double dose achieved the preset dose levels with high precision with median levels of 32.6 ng/ml, 66.4 ng/ml and 92.3 ng/ml respectively. 2/10 test persons had starting VD3 levels above 30 ng/ml, but 6 of the remaining 8 individuals showed a significantly increased ADCC after VD3 substitution to > 30 ng/ml. Further substitution to 65 ng/ml significantly increased ADCC activity in all 10 persons. 8/10 individuals were further substituted to achieve 90 ng/ml. However, in 7/8 of these probands the NK-cell-mediated ADCC was significantly reduced. The extent of the substitution-induced ADCC improvement varied individually and depended on the antibody concentration used to treat the target cells. Rituximab-mediated ADCC was significantly more affected by VD3 levels than obinutuzumab. Our study demonstrates that the ADCC of NK cells is optimal at median VD3 serum levels around 65 ng/ml and was chosen as the target VD3 level in the ongoing OPTIMAL>60 study of the German High-Grade Non-Hodgkin Lymphoma Study Group (DSHNHL) in elderly patients with DLBCL. This study is supported by the Eva Mayr-Stihl-Stiftung (Waiblingen, Germany). Disclosure: No conflict of interest disclosed. V716

A study of Myc-regulated metabolic changes in transformed B cells Jacobs L.A.1, Slawska J.1, Höllein A.1, Prehn C.2, Adamski J.2, Keller U.1 Klinikum rechts der Isar der Technischen Universität München, München, Germany, 2Helmholtz Zentrum München, München, Germany 1

consistent with deregulated cancer cell metabolism could be observed, supporting that therapeutic targeting of such pathways holds promise for treatment of aggressive cancers. Conclusion: These results provide novel insights into Myc-driven metabolism in transformed B-cells and may lead to exploitation of specific pathways to tackle a variety of Myc-mediated cancer cell liabilities. Disclosure: No conflict of interest disclosed. V717

B cell-specific conditional expression of Myd88 p.L252P leads to the development of diffuse large B cell lymphoma in mice Knittel G.1, Liedgens P.1, Korovkina D.1, Seeger J.M.2, Al-Baldawi Y.3, Al-Maarri M.4, Fritz C.1, Vlantis K.5, Bezhanova S.6, Scheel A.H.7, Wolz O.-O.8, Reimann M.9, Möller P.10, Lopéz C.11, Staudt L.M.12, Ortmann M.7, Pasparakis M.5, Siebert R.11, Schmitt C.A.9, Klatt A.R.13, Wunderlich F.T.4, Schäfer S.C.7, Persigehl T.3, Montesinos-Rongen M.14, Odenthal M.7, Büttner R.7, Frenzel L.P.1, Kashkar H.2, Reinhardt H.C.1 University Hospital of Cologne, Department I of Internal Medicine, Köln, Germany, 2University Hospital of Cologne, Institute for Microbiology and Hygiene, Cologne, Germany, 3University Hospital of Cologne, Department of Radiology, Cologne, Germany, 4Max-Planck-Institute for Metabolism Research, Cologne, Germany, 5University of Cologne, Cologne Excellence Cluster on Cellular Stress Response in Aging-Associated Diseases (CECAD), Cologne, Germany, 6N.N.Blokhin Russian Cancer Research Center, Moscow, Russian Federation, 7University Hospital of Cologne, Institute of Pathology, Cologne, Germany, 8University of Tübingen, Interfaculty Institute for Cell Biology, Department of Immunology, Tübingen, Germany, 9Charite – University Medical Center, Department of Hematology/Oncology, Berlin, Germany, 10Medical Faculty of the Ulm University, Institute of Pathology, Ulm, Germany, 11ChristianAlbrechts-University Kiel & University Hospital Schleswig Holstein, Institute for Human Genetics, Kiel, Germany, 12National Cancer Institute, NIH, Metabolism Branch, Center for Cancer Research, Bethesda, United States, 13University Hospital of Cologne, Institute for Clinical Chemistry, Cologne, Germany, 14 University Hospital of Cologne, Institute of Neuropathology, Cologne, Germany 1

Introduction: The transcription factor and proto-oncogene c-Myc (Myc) is well-established as activated across a broad range of cancers and is also known for its ability to drive malignant transformation of cells, including those of the hematopoietic system. Myc’s role in driving energy consumption and re-wiring cellular metabolism is also likely to contribute to the aggressive phenotype observed in diseases such as Burkitt’s lymphoma. Despite the apparent benefits of increased cellular metabolism in facilitating increased growth, proliferation and production of macromolecule precursors required for the rapidly dividing cell, the increased rate of metabolism can also render cells vulnerable to synthetic lethality approaches. As an example, dependency of Myc-driven cells upon glutaminolysis as a key energy and macromolecule precursor source renders cells glutamine-dependent and in the absence of glutamine cell death ensues. Additionally, as there is currently no clinical inhibitor of Myc, discovery of novel and targetable candidates is of significant value. Methods: To address this issue and further explore the role of oncogenic Myc in the deregulation of metabolism we performed targeted metabolic screens both in vitro and in vivo. Using the tetracycline-regulated conditional MYC gene in the P493-6 B-cell line we uncovered characteristic in vitro profiles of metabolites that change in response to Myc induction at several time points. Results: Altered profiles were observed for a variety of metabolites including biogenic amines and also for a number of amino acids. Functional testing of the metabolite candidates of interest confirmed altered levels of these metabolites in Myc-driven cells. Next we analyzed P493-6 xenografts in vivo. Metabolic imaging of the xenografts by means of FDG-PET demonstrated a reduction in tumour growth and glucose uptake upon Myc repression. Targeted metabolomic analysis revealed significant alterations in glucose utilisation and changes in metabolic profiles when Myc was repressed. Upon correlation of the in vitro and in vivo data, themes

The adaptor protein MYD88 is critical to relay activation of Toll-like receptor signaling to NF-κB activation. MYD88 mutations, particularly the p.L265P mutation, have been described in numerous distinct B cell malignancies, including diffuse large B cell lymphoma (DLBCL). 29% of activated B cell (ABC)-type DLBCL, which is characterized by constitutive activation of the NF-κB pathway, carry the p.L265P mutation. In addition, ABC-DLBCL frequently displays focal copy number gains affecting BCL2. We generated a novel mouse model (termed Myd88c-p.L252P), in which Cre-mediated recombination leads to the conditional expression of Myd88 p.L252P (the orthologous position of the human MYD88 p.L265P mutation) from the endogenous locus.  We activated our allele B cell-specifically by crossing it to the Cd19-Cre mouse. The resulting Myd88c-p. L252P/wt;Cd19Cre/wt animals develop splenomegaly and lymphadenopathy and succumb to a lymphoproliferative disease at a median of 500 days. Histological and immunohistochemical investigation revealed the sporadic emergence of large B cell lymphoma with an Bcl6-/Mum1+/ B220+/Cd138- immunotype. To potentially enhance lymphomagenesis in our mice, we aimed to combine the Myd88 p.L252P mutation with Bcl2 overexpression by making use of a newly generated R26LSL.BCL2 allele, where human BCL2 expression is driven by the CAGGs promoter. Indeed, all Myd88c-p.L252P/ wt;R26LSL.BCL2/wt;Cd19Cre/wt mice die of aggressive large B cell lymphoma at a median of 26 weeks. All animals showed splenomegaly and/ or lymphadenopathy, accompanied by infiltration of the liver. Clonality analysis by southern blot showed oligoclonality, suggesting a strong oncogenic potential of Myd88 p.L252P in combination with BCL2 overexpression. Interestingly, lymphoma cells were of a Bcl6-/Mum1+/B220-/ Cd138+ immunotype, in accordance with the plasmoblastic morphology that was histologically observed. In summary, we generated a mouse model that enables Cre-mediated expression of Myd88 p.L252P from the endogenous locus. Myd88c-p.L252P/ wt;Cd19Cre/wt mice develop and eventually die of lymphoproliferative

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disease. The emergence of diffuse large B cell lymphoma (DLBCL) was observed.  Combination of B cell-specific Myd88 p.L252P with BCL2 overexpression results in the development of an aggressive lymphoma with plasmoblastic features, most reminiscent of ABC-type DLBCL.

V719

Elucidation of tonic and activated B cell receptor signaling in Burkitt’s lymphoma reveals insights into non-oncogene addiction

Disclosure: No conflict of interest disclosed.

Walter R.1, Corso J.2, Pan K.-T.2, Doebele C.1, Yepes D.1, Sellner L.3, Tomska K.3, Bohnenberger H.4, Zenz T.3, Urlaub H.2, Serve H.1, Oellerich T.1

V718

HAX1 regulates cell surface expression and activity of CXCR4 in B-Cell Lymphoma Baumann U.1, Schmidt-Supprian M.1, Keller U.1,2, Peschel C.1,2, Bassermann F.1,2 Klinikum rechts der Isar der Technischen Universität München, München, Germany, 2German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), München, Germany 1

Introduction: HS-1-associated protein X-1 (HAX1) is a key regulator of cellular processes such as programmed cell death and migration, and has been implicated in B-cell development and survival. Recently, we characterized HAX1 as an oncogene in MCL that promotes lymphomagenesis and resistance to chemotherapy. Mechanisms of HAX1 deregulation include genomic deletion of its E3-ubiquitin-ligase FBXO25 or stabilizing phospho-degron mutations. C-X-C chemokine receptor 4 (CXCR4) is a major regulator of hematopoietic stem cell homing and migration and is overexpressed in various B-cell malignancies. WHIM-like mutations of CXCR4, derived from patients with Morbus Waldenström macroglobulemia, prevent the internalization of CXCR4 and induce constitutive downstream signaling, which has been linked to resistance to inhibitors of Brutons tyrosine kinase (BTK). The mechanism of how CXCR4 activity is regulated and CXCR4WHIM mutants are stabilized has remained unclear. Methods: Assays for interaction studies were performed by immunoprecipitations. Human and murine cell lines +/- shRNA constructs were treated with Plerixafor or Ibrutinib. Protein levels were determined by western blot, while CXCR4 surface expression was measured by flow cytometry. Cellular protein patterns were analyzed by proximity ligation assays and immunofluorescence. Results: Here we show that CXCR4 surface expression and activity are regulated by HAX1. CXCR4 and HAX1 directly interact in a BTK dependent manner. Treatment of cells with the CXCR4 antagonist Plerixafor leads to dissociation of the HAX1-CXCR4 complex, while CXCR4WHIM mutants remain associated with HAX1, thus revealing HAX1 as regulator of CXCR4 surface expression. Cells derived from Hax1 knock-out mice demonstrate nearly complete loss of Cxcr4 expression and unresponsiveness to the natural Cxcr4-ligand Sdf1. Moreover, re-expression of Hax1 in the Hax1-/- background restores Cxcr4 expression and activates Erk1/2 signaling. Studies in human B-NHL cells show that high expression levels of HAX1 promote resistance to Plerixafor, but also account for increased sensitivity to BTK inhibition. Moreover, knock-down of HAX1 in B-NHL cell lines induces an immediate reduction of CXCR4 surface expression and subsequent loss of overall CXCR4 levels, leading to increased Plerixafor sensitivity. Conclusion: We identify HAX1 as regulator of CXCR4 surface expression and activity, and suggest HAX1 as novel biomarker for response to CXCR4- and BTK-inhibition. Disclosure: No conflict of interest disclosed.

Goethe University Hospital, Department of Medicine II, Hematology/ Oncology, Frankfurt, Germany, 2Max Planck Institute for Biophysical Chemistry, Goettingen, Germany, 3Department of Translational Oncology, Heidelberg, Germany, 4Pathology, University Medical Center Göttingen, Goettingen, Germany

Burkitt’s lymphoma (BL) is treated with intensive chemotherapy in combination with anti-CD20 antibodies. Due to their toxicity, current treatment regimens are often not suitable for elderly patients and patients in developing countries where BL is endemic. BL cell survival was recently shown to rely on signals transduced by B cell antigen receptors (BCRs). However, tonic as well as activated BCR signaling networks and their functional relevance in BL remain elusive. Pharmacological interference with critical BCR signaling pathways might open up the possibility for targeted therapies in BL. In order (i) to elucidate BCR induced processes, (ii) to systematically compare tonic and activated BCR signaling and (iii) to identify drug targets in BL, we characterized BCR signaling in BL by quantitative phosphoproteomics. We identified and quantified about 16.000 phospho-sites in various BL cell lines, of which almost 1000 phospho-sites in about 600 proteins were regulated upon BCR stimulation as well as upon interference with tonic BCR signaling. The majority of the BCR signaling effectors identified has not been described in the context of B cell lymphomas yet. We identified commonalities between tonic and activated BCR signaling, but we also identified distinct events in tonic BCR signaling, including processes that are known to regulate survival signals. For instance, we found that phosphorylation of heat shock protein 90 (HSP90) is regulated by tonic BCR signaling. Inhibition of HSP90 by small molecule inhibitors such as AT13387 and STA-9090 as well as HSP90 knockdown induced apoptosis in most analyzed BL cell lines. Moreover, HSP90 inhibitors showed promising anti-tumor effects in BL xenotransplantation models. The induction of apoptosis by HSP90 inhibitors was caused by disruption of tonic BCR signaling that turned out to critically depend on HSP90 function as revealed by a phosphoproteomic pYome analysis. We found that upon BCR-dependent phosphorylation HSP90 is able to interact with important BCR effector proteins and that this interaction is crucial for their stabilization. Notably, expression of constitutively active forms of BCR-proximal kinases rendered BL cells resistant to HSP90 inhibitors. Taken together our study is a considerable complement to recent genetic studies that elucidated the mutational landscape in BL as it reveals mechanisms of addiction to non-oncogenes such as HSP90, ARFGEF2 and ACTN4. Disclosure: No conflict of interest disclosed. V720

LRPAP1, the predominant antigen of mantle cell lymphoma B cell receptors, induces proliferation and serves as a basis for a novel therapeutic strategy with ultimate specificity Thurner L.1, Hartmann S.2, Preuss K.-D.1, Bewarder M.1, Fadle N.1, Kemele M.1, Regitz E.1, Kim Y.-J.3, Pott C.4, Bohle R.-M.3, Hansmann M.-L.2, Pfreundschuh M.1 José Carreras Center for Immuno- and Gene Therapy and Internal Medicine I, Saarland University Medical School, Homburg/Saar, Germany, 2Dr. Senckenberg Institute of Pathology, Goethe University Hospital of Frankfurt Main, Frankfurt a. Main, Germany, 3Institute of Pathology, Saarland University Medical School, Homburg/Saar, Germany, 4Second Department of Medicine, University Hospital Schleswig-Holstein, Kiel, Germany 1

Background: Chronic antigenic stimulation may play an important role in the pathogenesis of malignant lymphomas. Although most MCL cas-

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es are believed to have an antigen-naive B cell as cell of origin, overrepresentation of certain VH genes and stereotyped CDR3s have been reported. Therefore we screened BCRs from MCLs for possible antigens. Methods: BCRs were expressed as recombinant Fabs based on corresponding pairs of functional variable region heavy and light chain genes, which had been amplified from isolated genomic DNA of snap-frozen MCL specimens and established MCL cell lines. The purified BCR-Fabs were checked for binding to proteins expressed on macroarrays of human cDNA expression libraries and on bacterial lysates. In addition, sera from patients with MCL were screened for antibodies against respective BCR antigens. Results: The recombinant MCL derived Fabs from 9 patients and of four established MCL cell lines were tested on proteinarrays. 4/9 of the recombinant MCL derived Fabs and Fabs derived of 1 of 4 of the stablished MCL cell lines reacted with human low density Lipoprotein Receptor-Related Protein associated Protein 1 (LRPAP1). Additionally we analyzed sera of patients with MCL and identified Anti-LRPAP1-autoantibodies with relevant titres in 8/28 patients. Anti-LRPAP1-antibodies were only detected in 1/200 sera of healthy controls. LRPAP1 induced proliferation, while a fusion protein of the LRPAP1 epitope and Pseudomonas exotoxin A induced specific apoptosis of a MCL cell line with a LRPAP1 reactive BCR (Maver1) without significantly affecting the proliferation of cell lines without reactive BCRs. Conclusion: LRPAP1 is the first identified frequent BCR target antigen of MCL. The high frequency of LRPAP1-reactive BCRs in MCL points strongly towards an important role of chronic autoreactive BCR stimulation against LRPAP1 in early steps of lymphomagenesis of MCL. Moreover transformed MCL cells with LRPAP1-reactive BCR still gain a growth advantage by the interaction with the cognate antigen suggesting, that LRPAP1 provides a proliferative environment for transformed MCL-cells with LRPAP1-reactive BCRs. The reason for the breakdown of self tolerance against LRPAP1 remains to be identified. Finally, retrograde targeting of LRPAP1-reactive MCL clones by administration of recombinant conjugates comprising the epitope of LRPAP1 and an immunotoxin, represents a new and highly specific therapeutic approach in MCL. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Chronische myeloische Leukämie – experimentell V723

Specific phosphorylation of Beclin 1 by BCR-ABL plays a crucial role in CML leukemogenesis by suppression of autophagy Yu C.1, Gorantla S.P.1, Müller T.1, Lippert L.J.1, Albers C.2, Huber T.3, Yue Z.4, Follo M.1, Duyster J.1, Illert A.L.1 Universitätsklinik Freiburg, Innere Medizin 1, Freiburg, Germany, 2Technical University Munich, III Medical Department, Munich, Germany, 3Universitätsklinik Freiburg, Renal Division, Freiburg, Germany, 4Mount Sinai School of Medicine, Departments of Neurology and Neuroscience, New York, United States 1

BCR-ABL tyrosine kinase inhibitors (TKIs) are used as standard treatment in chronic myelogenous leukemia (CML). However, CML stem cells are partly insensitive to TKI-induces cell death, and can thereby determine disease relapse. Autophagy, a genetically-regulated process of adaptation to metabolic stress, is involved in TKI-induced cell death and it has been hypothesized, that autophagy processes allow CML stem cells to become metabolically dormant thereby antagonizing TKI-induced cell death. However, the crucial molecular mechanisms of TKI-induced autophagy are poorly understood. Our study uncovers a crucial role of Beclin 1 phosphorylation in BCRABL-driven leukemia. We show, that active BCR-ABL suppresses autophagy by direct phosphorylation of Beclin 1 at tyrosine residues 233 and 352. Phosphorylated Beclin 1 negatively regulates autophagy by diminished binding of the positive regulators UVRAG, VPS15, ATG14 and

Abstracts

VPS34 and enhanced binding of the negative regulator Rubicon to the Beclin 1-complex, suggesting a critical role of Beclin 1 phosphorylation for complex stabilization and autophagy induction. Targeting Beclin 1 by a specific genetic approach in a murine bone morrow transplantation model substantiates the key role of the protein in BCR-ABL induced leukemia with significant prolonged survival of diseased Beclin 1-knockdown animals. Our findings highlight the importance of Beclin 1-mediated autophagy in BCR-ABL+ leukemia and provide a novel and promising treatment strategy for resistant or intolerant CML patients. Disclosure: No conflict of interest disclosed. V724

Impact of the Indoleamine 2,3-dioxygenase (IDO) system and its metabolites on molecular response to Nilotinib therapy in early Chronic Phase Chronic Myelogenous Leukemia (CML-CP) Sopper S.1, Mustjoki S.2, Gastl G.1, Trajanoski Z.3, Giles F.4, Hochhaus A.5, Janssen J.6, Geissler S.7, Fuchs D.7, Wolf D.8 Department of Hematology and Oncology, Innsbruck Medical University, Innsbruck, Austria, 2UCL Cancer Institute, Hematology Research Unit, University of Helsinki and Department of Hematology, Helsinki University Hospital Comprehensive Cancer Center, Helsinki, Finland, 3Divison for Bioinformatics, Biocenter, Innsbruck, Austria, 4NMDTI, Northwestern University, Chicago, United States, 5Klinik für Innere Medizin II, Universitätsklinikum, Jena, Germany, 6 Department of Hematology, VU University Medical Center, Amsterdam, Netherlands, 7Division of Biological Chemistry, Biocenter, Innsbruck Medical University, Innsbruck, Austria, 8Universitätsklinikum, Bonn, Germany 1

Introduction: Interferons induce the transcription of indoleamine 2,3 dioxygenase (IDO) in many cell types resulting in reduced T cell activation and proliferation by depletion of the essential amino acid tryptophan as well as by favoring immunosuppressive Treg expansion. Deregulation of IDO activity is linked to cancer immune evasion and the pathogenesis of autoimmune diseases. The role of IDO in CML remains elusive so far. Methods: A large panel of circulating pro-inflammatory cytokines and components of the IDO-pathway (soluble IDO=sIDO and kynurenine/ tryptophan ratio=KYN/TRP as a product of IDO activity) were analyzed alongside the prospective pan-european ENEST1st clinical study (NCT01061177). This substudy included 52 nilotinib-naïve Chronic Phase (CP)-CML patients that were subsequently treated with 300 mg BID nilotinib and analyzed at months 6 and 12. Molecular data were determined in central EUTOS reference laboratories. Results: Soluble IDO (sIDO) levels and KYN/TRP ratio are significantly up-regulated at diagnosis of CML and drop during nilotinib therapy. sIDO levels significantly correlate with increased KYN/TRP, suggesting increased IDO activity at diagnosis. Increased sIDO is linked to a pro-inflammatory status in CML patients, as it positively correlates to increased serum neopterin levels as well as various other pro-inflammatory markers, such as IFN-g, IL-8, IL-10, IL-17A, sVEGF-A, sVCAM-1 and sTNFR-1. Interestingly, as IDO is an IFN-regulated gene, we focused on the endogenous IFN-a producing plasmacytoid dendritic cells (pDC), which negatively correlated to KYN/TRP. A higher KYN/TRP is linked to superior molecular response, as demonstrated by a significant correlation to BCR-ABL/ABLIS levels. Patients having a high KYN/TRP ratio (> mean +2SD of post therapy levels) attain significantly faster and higher deep molecular response (i.e. MR4.5) rates. Conclusions: CML diagnosis in CP is linked to an inceased inflammatory status, as shown by increased levels of sIDO and its metabolites kynurenine leading to an increased KYN/TRP ratio. In solid cancer increased IDO expression/activity is linked to inferior outcome by favoring immune evasion. In contrast, in CML an increased KYN/TRP ratio is linked to improved molecular outcome during nilotinib 1st-line therapy. One reason could be that IDO may reflect endogenous IFN-α προδυλτιον, α known factor favoring immune-mediated CML-control. Disclosure: Sieghart Sopper: Expert Testimony: Novartis Dominik Wolf: Advisory Role: Novartis; Financing of Scientific Research: Novartis

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V725

V726

Comparison of survival between patients with and without switch to second-line tyrosine kinase inhibitor in a trial on chronic myeloid leukaemia with randomisation to first-line imatinib

Assessment of molecular response in CML patients with atypical BCR-ABL1 transcripts. Recommendations by the EUTOS collaboration

Pfirrmann M.1, Hasford J.1, Saussele S.2, Baerlocher G.M.3, Krause S.4, Hochhaus A.5, Hehlmann R.2, Lauseker M.1 Institut für Medizinische Informationsverarbeitung, Biometrie und Epidemiologie, Ludwig-Maximilians-Universität, München, Germany, 2 Medizinische Fakultät Mannheim, Universität Heidelberg, III. Medizinische Klinik, Mannheim, Germany, 3Universitätsklinik für Hämatologie, Inselspital, Bern, Switzerland, 4Medizinische Klinik 5, Universitätsklinik, Erlangen, Germany, 5 Klinik für Innere Medizin II, Universitätsklinikum, Jena, Germany 1

Introduction: In the German chronic myeloid leukaemia (CML) study IV, 1.408 patients were randomized to treatments starting with imatinib. During the course of the study, 342 switched to a second-line tyrosine kinase inhibitor (TKI). Aim of this analysis was to investigate the influence of treatment switch on survival. Methods: Patients in chronic phase with standard transcript types were considered, if actually receiving imatinib within half a year after diagnosis. The latest result on molecular response status within 3 months before switching while still in chronic phase as well as data on prognostic factors included in any established clinical score had to be known. Using a propensity score method, among non-switching patients, a matching partner was sought for each switching patient. To identify matching candidates, the influence of the prognostic factors on the probabilities of switching was estimated with the Fine-Gray model, considering progression and death as competing risks. The propensity score evaluated from the FineGray model had to be in the range of 0.2 times the standard deviation of all propensity scores. In addition, an appropriate matching candidate had still to be in first-line treatment at the time of the switch of his switching partner and the molecular remission status had to be the same. This supported the identification of two samples with similar switching probabilities and thus, similar distributions of prognostic factors. Results: In total, 150 switchers were eligible for the analysis. Median time to switch was 30 months after imatinib treatment start [range: 2-127]. Matching partners were identified for 143 switchers. Due to matching, in both samples, 77% of patients were without major molecular remission (MMR), 15% with MMR, and 8% better than MMR, showing the expected association between unsatisfactory remission and the inclination to switch. The hazard ratio for dying after matched “switching” times was 1.8 in case of switching [95%-confidence interval: 0.6-5.3], p = 0.3. Conclusions: Propensity score matching enabled finding two samples with similar distributions of prognostic variables and thus fostered comparability. Selection bias for switching but also power was reduced. There was no hint at possible survival differences between switchers and non-switchers necessitating immediate clinical action in the study when patients are still in chronic phase. A prospectively designed study on switching would be worthwhile. Disclosure: No conflict of interest disclosed.

Schaefer V.1, Ernst T.1, Saussele S.2, Lange T.3, Gerrard G.4, Cross N.C.P.5, Hochhaus A.6 Klinik für Innere Medizin II, Universitätsklinikum, Jena, Germany, 2III. Medizinische Klinik, Universitätsmedizin, Mannheim, Germany, 3Klinik für Hämatologie und internistische Onkologie, Asklepios-Klinikum, Weissenfels, Germany, 4Imperial Molecular Pathology, Hammersmith Hospital,, London, United Kingdom, 5Wessex Regional Genetics Laboratory, Salisbury District Hospital, Salisbury, United Kingdom, 6Universitätsklinikum Jena, Abt. Hämatologie/Onkologie, Jena, Germany 1

Introduction: Approximately 2‐3% of CML patients (pts) have variant BCR‐ABL fusions that cannot be monitored by RQ‐PCR using standard methodologies. Most rare variants are accounted for by seven variant fusions (e1a2/3, b2/3a3, e6a2, e8a2, e19a2). Within the European Treatment and Outcome Study (EUTOS) we sought to establish and validate robust RQ-PCR methods for each of these abnormalities. Methods: RNA was extracted from leukocytes obtained from 20 mL peripheral blood. Samples were received either locally or by mail and spent between 1 to 3 days in transit. Multiplex reverse transcription polymerase chain reaction (RT-PCR) experiments were performed to identify BCR-ABL1 transcripts (Cross et al., Leukemia 1994). In all cases, gel electrophoresis showed an unusual band. PCR products were then sequenced to characterize BCR-ABL1 rearrangements. Monitoring of residual disease was carried out by RQ-PCR with specific forward and/or reverse primers and probes using a serial dilution of individual BCR-ABL or GUSB plasmid DNA calibrators. GUSB transcripts were quantified as internal control and results were expressed as the ratio BCR-ABL1/GUSB with no conversion according to the international scale (IS) as it is recommended for b2a2 or b3a2 BCR-ABL1 only and cannot be applied to other transcripts in CML. Results: 43 pts from 8 prospective studies and pts outside of studies were investigated. A total of 435 samples (1-72 per patient, median 8) were analyzed. Puts expressed 5 different atypical BCR-ABL1 transcripts (e19a2, n = 18; e1a2, n = 10; b2a3, n = 5; b3a3, n = 5; b2a3&b3a3, n = 2, e8a2, n = 3). At the start of the observation period, ratios BCR-ABL1/ GUSB ranged between 0.01% and 179.1% (median 13.8%). Follow-up results were determined in comparison to the starting level and expressed as relative log reduction. Nine pts were in complete molecular remission considering RT-PCR negativity with a minimum GUSB transcript level of 20,000/reaction. During the observation period, 6 pts relapsed with a significant increase of the ratio BCR-ABL1/GUSB. Conclusions: Very few CML pts are diagnosed with unusual BCR-ABL1 transcripts, but the characterization of these rare BCR-ABL1 fusions is absolutely necessary for proper assessment of treatment response in the presence of these rare transcripts. Based on the individual PCR strategies, results cannot be expressed on the IS. The common molecular milestones and triggers for treatment discontinuation should not be applied. Disclosure: Vivien Schaefer: No conflict of interest disclosed. Andreas Hochhaus: Financing of Scientific Research: Novartis, BMS, Pfizer, Ariad; Expert Testimony: Novartis, BMS, Pfizer, Ariad

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V727

V728

The use of the EAC primers in digital PCR for monitoring minimal residual disease in CML generates background noise and requires a cut-off

The 3-month velocity of BCR-ABL decline: a tool for predicting deep molecular response in CML patients

Maier J.1, Franke G.-N.1, Cross M.1, Wildenberger K.1, Niederwieser D.1, Lange T.1,2 Universität Leipzig AöR, Leipzig, Germany, 2Asklepios Klinikum Weißenfels, Weißenfels, Germany 1

Background: Digital PCR (dPCR) is a highly sensitive method for the quantification of minimal residual disease and particularly relevant for monitoring response to treatment in CML. Aims: To define the technical limitations of BCR-ABL dPCR using primers and probes according to the Europe Against Cancer (EAC) protocol for monitoring deep molecular response in CML. Methods: The limit of detection was determined by analysing ten-fold dilutions of BCR-ABL from 100.000 to 1 copy in a background of wild type ABL. Specificity was determined in 3 independent runs of wild type (wt, Ba/F3 cDNA) and non template control (NTC, aqua dest) dPCR reactions. The sensitivity for BCR-ABL was determined in 5 dilutions of BCRABL positive cDNA from K562 cells (0.25, 2.5, 5, 10, 25 BCR-ABL copies per assay) in wild type ABL using a cut-off of 3 droplets. Results: The detection limit for BCR-ABL was 1 copy per assay, which was detected in 5/10 replicates. BCR-ABL to ABL dPCR was linear up to 120.000 copies/assay. Using a cut-off of >=1 positive drop, the specificity of BCR-ABL dPCR ranged from 78% to 93% (mean 85%) for NTCs and 71% to 89% (mean 79%) for wt DNA. The mean false-positive-rate was 15% (range 7-22%) for NTCs and 21% (range 11-29%) for wt. Increasing the cut-off resulted in specificities of 97% for >=2 droplets and 100% for >=3 droplets. Therefore the limit of quantitation (LOQ) was defined as a minimum of 3 positive droplets for sample positivity. For ABL, a cut-off of >=1 positive droplet resulted in a high specificity of 98%. A sensitivity of 100% was obtained for dilutions of 25, 10 and 5 copies per replicate when assessing the samples in duplicates using the cut-off of 3 positive droplets, even though 43% of the replicates containing 5 copies tested negative. 2.5 and 0.25 copies were detected in 79% and 13% of the samples when using duplicates, emphasizing the importance of the LOQ of 3 copies predicted by the Poisson distribution. Summary/Conclusion: 1) The M-BCR-ABL primers in widespread use in the EAC dPCR protocol are suboptimal, since the 3 droplet cut-off required to avoid false positives results in a loss of sensitivity. 2) The FPR observed in BCR-ABL dPCR seems to be assay specific. Further optimization of the primer/probe system is currently underway to enable reliable assessment of deep molecular responses in CML patients by dPCR. 3) Sensitivity can be greatly improved by assessing more replicates with a primer/probe system without background. Disclosure: No conflict of interest disclosed.

Rinaldetti S.1, Lauseker M.2, Fabarius A.1, Hehlmann R.1, Khaled N.3, Dietz C.T.4, Proetel U.1, Krause S.5, Schubert J.6, Einsele H.7, Hänel M.8, Dengler J.9, Falge C.10, Kanz L.11, Neubauer A.12, Heydrich B.13, Stegelmann F.14, Pfreundschuh M.15, Waller C.F.16, Spiekermann K.17, Baerlocher G.M.18, Pfirrmann M.2, Hofmann W.-K.1, Hochhaus A.19, Müller M.C.4, Saußele S.1, for the SAKK and the German CML Study Group Medizinische Fakultät Mannheim, Universität Heidelberg, III. Medizinische Klinik, Mannheim, Germany, 2Ludwig-Maximilians-Universität, Institut für Medizinische Informationsverarbeitung, Biometrie und Epidemiologie, München, Germany, 3Medizinische Fakultät Mannheim der Universität Heidelberg, III. Medizinische Klinik, Hämatologie und Onkologie, Mannheim, Germany, 4IHO Institute for Hematology and Oncology, Mannheim, Germany, 5Universitätsklinikum Erlangen, Medizinische Klinik 5, Erlangen, Germany, 6Elblandklinikum Riesa, Klinik für Innere Medizin II, Riesa, Germany, 7 Universitätsklinikum Würzburg, Medizinischen Klinik und Poliklinik II, Würzburg, Germany, 8Klinikum Chemnitz, Klinik für Innere Medizin III, Chemnitz, Germany, 9Onkologische Schwerpunktpraxis, Heilbronn, Germany, 10Klinikum Nürnberg Nord, Ambulantes Behandlungszentrum, Onkologische Praxis, Nürnberg, Germany, 11Universitätsklinikum Tübingen, Medizinische Universitätsklinik, Innere Medizin II, Tübingen, Germany, 12 Universitätsklinikum Gießen und Marburg, Standort Marburg, Philipps Universität, Klinik für Hämatologie/Onkologie und Immunologie, Marburg, Germany, 13Universitätsklinikum Schleswig-Holstein, Klinik für Innere Medizin II, Kiel, Germany, 14Universitätsklinikum Ulm, Klinik für Innere Medizin III, Ulm, Germany, 15Universitätsklinikum des Saarlandes, Klinik für Innere Medizin I, Homburg, Germany, 16Universitätsklinikum Freiburg, Klinik für Innere Medizin I, Freiburg, Germany, 17Klinikum der Universität München, Campus Großhadern, Medizinische Klinik und Poliklinik III, München, Germany, 18Inselspital, Universitätsklinik für Hämatologie und Hämatologisches Zentrallabor, Bern, Switzerland, 19Universitätsklinikum Jena, Abteilung Hämatologie/Onkologie, Klinik für Innere Medizin II, Jena, Germany 1

Introduction: Rationales for the prediction of molecular response gain interest as deep molecular response (DMR) is increasingly used for subsequent therapy discontinuation. Here we correlate the 3-month velocity of BCR-ABL decline with DMR (MR4 and MR4.5 as defined by Cross, Leukemia, 2015) over a follow-up period of 5 years. In a further step, we address the question whether ABL may also be used as a reference gene (RG). Methods: The 3-month velocity was calculated for a cohort of 239 treatment-naïve patients from the CML IV study using GUS as reference genes. Gene expression analysis has been performed by qRT-PCR and the velocity was calculated as described previously (Hanfstein, Leukemia, 2014). Cumulative incidence (CI) curves were estimated for the achievement of DMR considering death and disease progression as competing risks and compared using the Gray test over a follow-up period of 5 years. Probabilities of overall survival (OS) and progression free survival (PFS) were estimated by the Kaplan-Meier method over a follow-up period of 8 years. In parallel, BCR-ABL/ABL ratios were calculated at diagnosis. Results: The 3-month velocity calculated with GUS as RG showed a distinct cut-off for a 3 month reduction ratio of 0.35 permitting a risk stratification for OS (p < 0.001, 8-year OS: 77% vs 96%) and for PFS (p < 0.001, 8-year PFS: 78% vs 96%). In addition, the velocity of BCR-ABL decline yielded CI curves that discriminate between 2 groups with significantly different probabilities of achieving MR4 (p < 0.001, 5-years MR4: 51% vs 79%) and MR4.5 (p < 0.001, 5-years MR4.5: 33% vs 62%). We show that the distribution of BCR-ABL/ABL levels at diagnosis forms a plateau over an expression of 10%, so that the determination of the velocity with ABL, by means of response prediction, may be questionable. Conclusion: The 3-months velocity calculated with GUS as RG is also a critical prognostic discriminator for DMR. In contrast to the RG GUS, the BCR-ABL/ABL quotient showed no linearity at time of CML diagnosis because the ABL amplicon is also included in the BCR-ABL transcript. Disclosure: Sébastien Rinaldetti: No conflict of interest disclosed. Susanne Saußele: Financing of Scientific Research: Novartis, BMS, Pfizer, ARIAD; Expert Testimony: Novartis, BMS

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Freier Vortrag

V730

Multiples Myelom experimentell V729

Immunomodulatory drugs mediate both anti-tumor activity and teratotoxocity via destabilization of CD147 and MCT1 Eichner R.1, Heider M.1, Fernández-Sáiz V.1,2, van Bebber F.3, Garz A.-K.1,2, Lemeer S.4, Rudelius M.5, Targosz B.-S.1, Jacobs L.1, Platzbecker U.6, Germing U.7, Langer C.8, Knop S.9, Einsele H.9, Peschel C.1, Keller U.1, Schmid B.3, Götze K.S.1,2, Küster B.2,4,10, Bassermann F.1,2 III. Medizinische Klinik, Klinikum rechts der Isar, München, Germany, German Cancer Consortium (DKTK), Heidelberg, Germany, 3German Center for Neurodegenerative Diseases (DZNE), München, Germany, 4Department of Proteomics and Bioanalytics, Technische Universität München, Freising, Germany, 5Institute of Pathology and Comprehensive Cancer Center Mainfranken, Würzburg, Germany, 6Medizinische Klinik und Poliklinik 1, Medizinische Fakultät Carl Gustav Carus, TU Dresden, Dresden, Germany, 7 Department of Hematology, Oncology and Clinical Immunology, HeinrichHeine University, Düsseldorf, Germany, 8Department of Internal Medicine III, University of Ulm, Ulm, Germany, 9Division of Hematology and Medical Oncology, Department of Internal Medicine, University Medical Center, Würzburg, Germany, 10Center for Integrated Protein Science Munich, Freising, Germany 1 2

Introduction: Immunomodulatory drugs (IMiDs) as thalidomide, lenalidomide and pomalidomide, have immunomodulatory, anti-proliferative, anti-angiogenic and teratotoxic effects and are key treatment components for hematologic malignancies such as multiple myeloma (MM) and del(5q) myelodysplastic syndrome (MDS). IMiDs bind to Cereblon (CRBN), a substrate receptor of CRL4 ubiquitin ligases, to mediate anti-cancer and teratotoxic effects. Recently, IKZF1/3 and CK1-alpha were identified as substrates, which are degraded upon IMiD-induced binding to CRBN. However, both the physiological function of CRBN, as well as a common mechanism by which IMiDs exert their versatile effects, remain largely elusive. Methods: Tandem-affinity purification followed by mass spectrometry identified CRBN interactors. Binding assays included immunoprecipitations, GST-pulldowns and chromatography experiments. MM and MDS cell lines were treated with IMiDs or shRNA constructs, and proliferation, lactate levels and VEGF/MMP7 secretion was evaluated. Patient-derived primary MM and MDS cells were treated with IMiDs in-vitro. Protein expression was analyzed by western blot and flow cytometry. For teratotoxicity studies, zebrafish larvae were treated with IMiDs or morpholinos targeting CD147. Results: Here we identify a novel, ubiquitin-independent physiological chaperone-like function of CRBN, which promotes maturation of CD147 and MCT. The CD147/MCT1 transmembrane complex is involved in various biological functions including angiogenesis, proliferation, invasion and lactate export. We found that IMiDs outcompete CRBN binding to CD147/MCT1, leading to CD147/MCT1 destabilization and decreased proliferation, lactate export and VEGF/MMP7 secretion. Both IMiD-sensitive MM cell lines and primary patient-derived MM cells lose CD147/ MCT1 expression upon IMiD exposure, while resistant cells retain expression. Furthermore, del(5q) MDS cells demonstrate elevated CD147/ MCT1 expression levels, which are attenuated to the levels of non-del(5q) MDS cells upon IMiD treatment. Finally, thalidomide destabilizes CD147 in zebrafish, a common teratotoxicity model, and knockdown of CD147 fully phenocopies the teratotoxic effects of thalidomide in a dose-dependent manner. Conclusion: We describe a novel chaperone-like function of CRBN, distinguish CD147 and MCT1 as major mediators of IMiD function and provide a common mechanism explaining both the anti-tumor and the teratotoxic effects of IMiDs.

Elotuzumab plus lenalidomide and dexamethasone (ELd) in relapsed/refractory multiple myeloma (RRMM): ELOQUENT-2 post-hoc analysis of progression-free survival (PFS), tumor regrowth by time from diagnosis and prior lines of therapy, and median duration of response (DOR) Weisel K.1, Dimopoulos M.A.2, Palumbo A.3, Richardson P.4, Mateos M.-V.5, Moreau P.6, Gupta M.7, Sheng J.7, Passey C.7, Sy O.7, Katz J.7, Lonial S.8 University of Tubingen, Tubingen, Germany, 2National and Kapodistrian University of Athens, Alexandra Hospital, Athens, Greece, 3A.O.U. San Giovanni Battista di Torino – Ospedale Molinette, Torino, Italy, 4Dana-Farber Cancer Institute, Boston, United States, 5Complejo Asistencial Universitario de Salamanca-IBSAL, Salamanca, Spain, 6University Hospital, Nantes, France, 7 Bristol-Myers Squibb, Princeton, United States, 8Winship Cancer Institute, Emory University, Atlanta, United States 1

Introduction: ELOQUENT-2 (NCT01239797) is a ph 3 randomized study to assess efficacy/safety of ELd vs lenalidomide/dexamethasone (Ld) in patients (pts) with RRMM. Median time from diagnosis was 3.5y. 3y follow-up showed 27% reduction in risk of disease progression/death with ELd (HR 0.73; 95% CI 0.60−0.89; p = 0.0014), with durable efficacy across key subgroups.1 There was a 1y delay in time to next treatment with ELd vs Ld: median (95% CI) of 33 (26.2−40.2) mo vs 21 (18.1−23.2) mo (HR 0.62; 95% CI 0.50−0.77).1 Interim overall survival (OS) results showed a trend toward long-term benefit of ELd vs Ld. Methods: RRMM pts with 1−3 prior therapies were randomized to ELd or Ld in 28-day cycles until disease progression/unacceptable toxicity. Coprimary endpoint: PFS. Exploratory analyses assessed effect of time from diagnosis and number of prior lines of therapy on PFS. Dynamic modelling evaluated rate of serum M-protein suppression and tumor regrowth. Results: 646 pts were randomized (ELd, 321; Ld, 325). At baseline median age was 66y; 32% of pts had del(17p), 9% t(4;14); median 2 prior therapies; 35% of pts were refractory to last therapy. Median DOR: 20.3 mo with ELd vs 16.6 mo with Ld. In pts with ≥median time from diagnosis, median PFS was 26.0 mo with ELd vs 17.3 mo with Ld (HR 0.60; p = 0.0004). Pts in the ELd group with ≥median time from diagnosis and 1 prior line of therapy (n = 48) had 53% reduction in the risk of progression/death (HR 0.47; p = 0.013) vs Ld group (n = 61). PFS HR in pts with ≥median time from diagnosis and >1 prior line of therapy was 0.59 (p = 0.004; ELd n = 111; Ld n = 96). Serum M-protein dynamic modelling (n = 629) showed slower regrowth rate for pts in the ELd vs Ld group regardless of time since diagnosis. Slowest rate (g/dL.day-1) of tumor regrowth (0.00145) was in pts in the ELd group with ≥median time from diagnosis (vs Ld, 0.00234; < median time from diagnosis: ELd, 0.00279; Ld, 0.00389). Conclusions: ELOQUENT-2 shows PFS benefit with ELd vs Ld across key subgroups. In this post-hoc analysis, PFS benefit was seen with ELd vs Ld across key subgroups by time from diagnosis and number of prior therapies. This is supported by serum M-protein dynamic modelling, which suggests that ELd may differentially slow tumor regrowth rate, providing sustainable and long-term survival benefits as an immuno-oncology agent. Further analyses to identify pts with increased PFS and OS benefit from elotuzumab are ongoing. Reference: 1 Dimopoulos M et al.: ASH 2015;Oral 28. Disclosure: Katja Weisel: Advisory Role: Bristol-Myers Squibb, Celgene, Amgen, Onyx, Novartis, Takeda, Janssen; Financing of Scientific Research: Bristol-Myers Squibb, Celgene, Amgen, Onyx, Janssen, Novartis, Takeda; Expert Testimony: Janssen, Celgene Sagar Lonial: Advisory Role: Millennium, Celgene, Novartis, Bristol-Myers Squibb, Janssen, Onyx; Expert Testimony: Millennium, Celgene, Novartis, Bristol-Myers Squibb, Janssen, Onyx

Disclosure: Ruth Eichner: No conflict of interest disclosed. Florian Bassermann: Expert Testimony: Celgene

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V731

Bone marrow mesenchymal stromal cells of multiple myeloma patients are characterized by metabolic aberrations during mitochondrial stress Berenstein R.1, Waechter M.1, Nogai A.1, Blau O.1, Kunitz A.1, Pezzutto A.1, Doerken B.1, Blau I.W.1 Charité University Berlin, Department of Hematology, Oncology and Tumorimmunology, Berlin, Germany 1

Introduction: Proliferation of multiple myeloma (MM) cells is strongly depending on interaction with bone marrow mesenchymal stromal cells (BMMSCs). In this study, we investigated the role of NAD-dependent deacetylase sirtuin-3 (SIRT3) in BMMSCs and analyzed effect of monocarboxylate transporter (MCT) on SIRT3 expression in BMMSCs obtained from myeloma patients (MM-BMMSCs). Methods: BMMSCs from healthy donors (HD-BMMSCs) were transfected to transient knock-down of SIRT3 using siRNAs (Qiagen). Mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) were analyzed by FACS. Cell cycle analysis was done using flow cytometry cell cycle analysis assay (Abcam) and senescence was examined using FACS and senescence-associated β-galactosidase activity. For apoptosis, Annexin V-FITC Kit (Miltenyi) was used. Protein expression was examined by western blot. The effect of MCT transporter interaction was done using α-cyano-4-hydroxycinnamic acid. Results: SIRT3 knock-down in HD-BMMSCs induced 1.4 - 1.9-fold increasing of ROS levels (p < 0.05). This was associated with dissipation of MMP (1.4-fold - 1.8-fold, depending on type of siRNA) (p < 0.04). Inhibition of SIRT3 created cell cycle arrest in S phase in MM-BMMSCs. Percentage of BMMSCs in S phase increased upon SIRT3 knock-down between 6.7%-9.6% (p < 0.039). Reduction of SIRT3 led to 1.5-fold increased senescence-associated β-galactosidase activity in transfected HDBMMSCs indicating that this protein could be responsible for premature senescence of BMMSCs of myeloma patients (p < 0.03). Interaction of BMMSCs with MM cells via MCT transporters could influence SIRT3 levels in BMMSCs of MM patients. Inhibition of MCT interaction induced apoptosis in MM cells but not BMMSCs (p < 0.04). Furthermore, suppression of MCT interaction between MM cells and MM-BMMSCs reduced activation of SIRT3 around 1.7-fold in MM-BMMSCs. Conclusion: Reduced expression of SIRT3 in MM-BMMSCs could be the reason for increased ROS levels, cell cycle arrest in S phase and premature senescence-like state of MM-BMMSCs. Inhibition of MCT transporters in MM cells and MM-BMMSCs induces apoptosis in MM cells and inhibits increased expression of SIRT3 in MM-BMMSCs. Disabling of MCT transporter interaction could possibly inhibit sustained induction of mitochondrial stress response in MM-BMMSCs and circumvent induction of the premature senescence-like state. SIRT3 seems to be a major regulator of mitochondrial functions in BMMSCs. Disclosure: Rimma Berenstein: No conflict of interest disclosed. Igor Wolfgang Blau: Expert Testimony: Celgene V732

Notch/miR-223 modified VEGF/IL-6 Levels and osteogenic potential in multiple myeloma bone marrow stromal cells Berenstein R.1, Waechter M.1, Blau O.1, Nogai A.1, Kunitz A.1, Pezzutto A.1, Doerken B.1, Blau I.W.1 Charité University Berlin, Department of Hematology, Oncology and Tumorimmunology, Berlin, Germany 1

Introduction: Bone marrow mesenchymal stromal cells (BMMSCs) represent a crucial component of multiple myeloma (MM) microenvironment supporting its progression and proliferation. Recently, microRNAs have become an important point of interest for research on micro-environmental interactions in MM with some evidence of tumor supportive roles in MM. Methods: In this study, we examined the role of miR-223 for MM support in BMMSCs of 56 patients with MM (MM-BMMSCs).

Abstracts

Results: miR-223 expression in MM-BMMSCs was reduced by the presence of MM cells in vitro in a cell-contact dependent manner compared to mono-cultured MM-BMMSCs. Co-cultivation of MM cells and MMBMMSCs induced activation of notch amongst others via jagged-2/ notch-2 leading to increased expression of Hes1, Hey2, or Hes5 in both cell types. Cultivation of MM-BMMSCs with increasing levels of recombinant jagged-2 reduced miR-223 and increased Hes1 levels in a concentration-dependent manner. Transient reduction of miR-223 levels increased VEGF and IL-6 expression and secretion by MM-BMMSCs. In addition, reduction of miR-223 degraded the osteogenic differentiation potential of MM-BMMSCs. Inhibition of notch signaling induced apoptosis in both MM cells and MM-BMMSCs. Furthermore, it increased miR-223 levels and reduced expression of VEGF and IL-6 by both cell types. Conclusion: These data provide first evidence that miR-223 participates in different MM supporting pathways in MM-BMMSCs including regulation of cytokine secretion and expression as well as osteogenic differentiation of MM-BMMSCs. More insights on the role of miR-223 in MM-BMMSCs and in cellular interactions between MM cells and MMBMMSCs could provide starting points for a more efficient anti-myeloma treatment by targeting of notch signaling Disclosure: Rimma Berenstein: No conflict of interest disclosed. Igor Wolfgang Blau: Expert Testimony: Grant Celgene V733

PIM1-inhibition enhances and prolongs Plerixafor-induced mobilization of HSCs Müller T.A.1, Zwick A.2, Decker S.1, Klein C.1, Rister B.1, Kissel S.1, Follo M.1, Wäsch R.1, Illert A.L.1,3, Dierks C.1, Duyster J.1,3 Universitätsklinik Freiburg, I. Medizinische Klinik, Freiburg, Germany, 2III. Medizinische Klinik (Hämatologie/Onkologie), Klinikum rechts der Isar, Technische Universität München, München, Germany, 3Deutsches Konsortium für Translationale Krebsforschung (DKTK), Heidelberg, Germany 1

Introduction: The CXCL12/CXCR4 axis was shown to be a major regulator for the interaction of hematopoietic stem cells (HSCs) with the niche and interruption of this pathway mobilizes HSCs from the bone marrow (BM). Therefore CXCR4 antagonists like AMD3100 (Plerixafor) are used in the clinic for the collection of HSCs from patients who fail to mobilize HSCs in response to G-CSF. In our previous studies, we could show that the serine/threonine kinase PIM1 is regulating the surface expression of the CXCR4 receptor on HSCs. In addition, PIM1-deficient HSCs fail to home to a wild type (WT) BM niche. Based on these results, we aimed to improve HSC mobilization by combining CXCR4 and PIM inhibition. Methods: In order to study the mobilization efficiency in the murine model, mice were treated with AMD3100 alone or in combination with LGB321, a novel pan-PIM inhibitor. The mice were sacrificed at various timepoints and peripheral blood (PB) was isolated. The percentage of HSCs was then determined by flow cytometry. The mechanism of HSC mobilization was studied in isolated HSC and stroma populations by analyzing mRNA levels and surface expression of CXCR4, its ligand CXCL12 and other factors, which are crucial for the interaction of HSCs and stromal cells. Results: We found that CXCR4 inhibition using AMD3100 leads to a compensatory upregulation of CXCR4 surface expression on total BM cells as well as HSCs. This effect can be reverted by deficiency or inhibition of PIM1. As a consequence, HSC mobilization using AMD3100 is strongly enhanced and prolonged in Pim1-deficient mice compared to WT animals. Likewise, treatment of WT animals with AMD3100 in combination with LGB321 leads to increased and prolonged HSC mobilization compared to animals treated only with AMD3100. Besides the downregulation of CXCR4 on HSCs, we found that the Cxcl-12 expression as well as CXCR4 surface expression in CXCL12-abundant reticular (CAR) cells is dramatically decreased in Pim1-deficient mice, which even further increases the mobilization capacity of Pim1-deficient mice. Conclusion: Our findings indicate, that PIM1 inhibition counteracts the compensatory upregulation of the CXCR4 receptor on HSCs after Plerix-

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afor treatment and decreases CXCL12 levels within the bone marrow niche. Therefore, targeting PIM kinases in combination with CXCR4 inhibition could improve the collection of stem cells in patients at risk for poor mobilization. Disclosure: No conflict of interest disclosed. V734

Peripheral B cells from patients with MGUS and Multiple Myeloma (MM) can be stimulated by Paraprotein-target specific T-helper cells to produce Paraprotein-identical monoclonal antibodies: Rationale for PARs, a novel therapeutic approach with ultimate specificity in MGUS/MM Bewarder M.1, Kubuschok B.2, Preuss K.D.1, Schormann C.1, Pfreundschuh M.1, Neumann F.1 Universitätsklinikum des Saarlandes, Innere Medizin I, Homburg, Germany, Klinikum-Augsburg, II. Medizinische Klinik, Augsburg, Germany

1 2

Introduction: Paratarg-7 (P-7) is the paraprotein target from 15% of European and 37% of African-American MGUS/MM patients, supporting a role of P-7 in the pathogenesis of MGUS/MM via chronic auto-antigenic stimulation. All patients with such paraproteins are carriers of a hyperphosphorylated form of P-7 (pP-7). We identified pP-7-specific T-helper cells which were restricted by certain “permissive” HLA-DR haplotypes. These haplotypes are overrepresented among the corresponding patients compared to the normal population. Thus, there are two prerequisites for the development of MGUS/MM with a P-7-specific paraprotein: 1st carriership of pP-7 and 2nd a permissive HLA-DR subtype. We now investigated the interaction of pP-7-specific T-helper cells and peripheral B cells with cognate specificity. Methods: B cells of nine MGUS/MM patients with a P-7-specific paraprotein and from one healthy pP-7-carrying son of one of these patients were analyzed. For in vitro stimulation B and T cells were magnetically isolated from the PBMC. T cells were replaced by pP-7-specific T-helper cell clones. Results: In all patients pP-7-specific T-helper cells stimulated peripheral B cells to produce P-7-specific antibodies. These B-cell responses were monoclonal and the immunoglobulin type was identical with the paraprotein of the corresponding patient. B-cell stimulation with CMV-specific T-helper cells always induced an antigen-specific, yet polyclonal response. As against the diseased mother, the peripheral B cells of her pP-7-carrying healthy son secreted polyclonal P-7-specific antibodies. Conclusion: In patients with MGUS/MM peripheral B cells can be induced to produce monoclonal antibodies identical to the paraprotein by T-helper cells with specificity for the paraprotein target. This supports an indispensable role of these T-helper cells in the pathogenesis of MGUS/ MM via chronic antigenic stimulation. Our results prove that precursors of the malignant plasma cells can be found in the peripheral blood that might fuel the development of malignant plasma cells. These B cells can now be targeted by PARs (paraprotein antigens for reverse targeting) conjugated to toxins, as parts of bispecific constructs (PAR/CD3 or PAR/ CD16) or PAR/CAR T cells. PARs can be envisaged prophylactically for carriers of modified autoantigens with a permissive MHC-II haplotype and a monoclonal B-cell response in vitro or in MM patients achieving a VGPR or CR after treatment for relapse prevention. Disclosure: No conflict of interest disclosed.

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Wissenschaftliches Symposium Epigenetik myeloischer Neoplasien V737

Mutant IDH as therapeutic target in AML/MDS: where Epigenetics meets metabolomics Heuser M.1 Medizinische Hochschule Hannover, Hannover, Germany

Mutations in the metabolic enzymes isocitrate dehydrogenase 1 (IDH1) and isocitrate dehydrogenase 2 (IDH2) are frequently found in several tumors, including acute myeloid leukemia (AML), glioma, chondrosarcoma and intrahepatic cholangiocarcinoma. Clinical data show that inhibition of mutant IDH2 induces differentiation of blast cells. However, preliminary data indicate that the mutant allele burden remains high, suggesting that the leukemic stem cell is not depleted with current IDH inhibitors. Therefore, a closer look at the role of mutant IDH1/2, its oncometabolite 2-hydroxyglutarate (2HG) and their interplay with epigenetic regulation is warranted. Mutations in IDH1 and IDH2 occur almost exclusively at conserved amino acids that affect the active sites where IDH1/2 substrates isocitrate and NADP+ bind. The conversion of isocitrate to αKG is impaired in the mutant proteins, resulting in reduced production of αKG and NADPH. (11) Furthermore, mutant IDH1 proteins gain a neomorphic ability to convert αKG to R-2-hydroxyglutarate (R-2HG). Almost all patients with canonical IDH1/2 mutations express high levels of intracellular R-2HG, while an increase of the S-enantiomer of 2HG has not been described in AML, glioma, chondrosarcoma or intrahepatic cholangiocarcinoma patients. While αKG is a cofactor for many dioxygenases involved in diverse processes like chromatin modification, hypoxic response, nucleic acid repair and modification, and fatty acid metabolism, it was shown that 2HG is an inhibitor of these dioxygenases. Surprisingly, S-2HG inhibits most dioxygenases more potently than R-2HG. I will discuss recent findings on the role of R-2HG and S-2HG and its implications on the pathophysiology of mutant IDH1/2 proteins in AML. Disclosure: Michael Heuser: Honoraria: Der IDH1 Inhibitor HMS-101 wurde als Patent angemeldet.

Debatten

MDS meets MPN V738

MDS and MPN – a historical view on two disease groups Aul C.1, Germing U.2, Bennett J.M.3 HELIOS St. Johannes Klinik, für Hämatologie, Onkologie u. klinische Immunologie, Duisburg, Germany, 2Universitätsklinikum Düsseldorf, Klinik für Hämatologie, Onkologie u. klinische Immunologie, Düsseldorf, Germany, 3 University of Rochester Medical Center, Rochester, United States 1

Introduction: Precise definition and classification of haematological disorders are essential for correctly diagnosing, treating and studying patients with these entities. With the advent of new diagnostic tools e.g. cytogenetics, immunophenotyping and molecular analyses, tumours of haematological origin could be more effectively classified. In 1982, the world of the haematologist was relatively simple and almost entirely based on morphological techniques. Results and conclusions: Progress in the definition and categorization of haematological diseases primarily stemmed from conceptual innovations which created a new framework for further studies. Dameshek, Block and Bennett were the first to realize that patients with refractory anemia and smouldering leukemia belonged to the same group of diseases, namely the myelodysplastic syndromes (MDS). They also played a fundamental role for the definition and grouping of the chronic myeloproliferative diseases. In 1982, this disease group included 4 entities. The group of myelodysplastic/myeloproliferative neoplasms with overlapping clinical, laboratory and

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morphological features was not yet recognized, although it was already known that cases of myeloproliferative diseases without presence of the Philadelphia chromosome existed. The nosological position of the chronic myelomonocytic leukemia (CMML) which was included into the MDS category by the FAB group was controversely debated as it showed both myelodysplastic and myeloproliferative features (MDS/MPN). Besides CMML, this group of diseases now includes juvenile myelomonocytic leukemia (JMML), atypical chronic myeloid leukemia (aCML) and other disorders which are negatively defined by the absence of a BCR-ABL1 fusion gene or rearrangements of the PDGFRA, PDGFRB and FGFR1 genes. The clonal myeloid disorders which could be distinguished in 1982 are now included in various editions of the WHO classification which incorporates new morphological, immunophenotypical, cytogenic and molecular features which, in addition to a more precise definition of morphological categories, also carry relevant prognostic information. Disclosure: No conflict of interest disclosed. V739

Today´s view on MDS and MPN – common features and different interpretations Hofmann W.-K.1, Reiter A.1 Universitätsmedizin Mannheim, Hämatologie und Onkologie, Mannheim, Germany 1

Chronic myeloid neoplasms comprise MDS, MPN, MDS/MPN and myeloid/lymphoid neoplasms with eosinophilia and rearrangement of PDGFRA, PDGFRB or FGFR1 (MLN-eo). MPN include the “classical” subtypes CML, PV, ET, and PMF and the “rare” subtypes CNL, CELNOS and SM while MDS/MPN comprise CMML, atypical CML, MDS/ MPN unclassified and JMML. For MDS, a total of 10 different sub-entities are established including MDS with: single lineage dysplasia (MDSSLD), multilineage dysplasia (MDS-MLD), ring sideroblasts (MDS-RS), excess blasts (MDS-EB), and del(5q). For decades, the various subtypes have mainly been differentiated on basis of clinical and morphological characteristics. The identification of BCR-ABL in CML and more than 50 structurally related tyrosine kinase (TK) fusion genes in MLN-eo and MDS/MPN-eo and their successful treatment with tyrosine kinase inhibitors such as imatinib has opened an unprecedented era on the search for potential targets in other chronic myeloid neoplasms. Further milestones were set by the identification of mutations in JAK2, MPL and CALR in PV, ET and PMF and the subsequent development of JAK inhibitors. In the meantime, modern DNA/RNA-based high throughput sequencing technologies identified in MDS and MDS/MPN patients a huge number of point mutations or small deletions/insertions in a large variety of genes, e.g. epigenetic regulators, splicing machinery components, or transcription factors. However, it also became clear that the molecular pathogenesis is far more complex than expected as numbers and specific patterns of molecular aberrations differently influence the clinical course, response/ resistance to treatment and prognosis. Surprisingly, the same molecular aberrations, which were initially identified in MDS and MDS/MPN, have meanwhile also been identified in classical and rare MPN. In MPN, accumulating evidence suggests that mutations in JAK2 or KIT may mainly represent phenotype-modifying mutations while additional mutations, e.g. in ASXL1, SRSF2, EZH2, RUNX1 etc., and their number have a similar negative impact on prognosis like in MDS or MDS/MPN, irrespective of the clinical phenotype. In conclusion, recently identified molecular and disease-mechanistic parameters give evidence that a further discrimination of MPN and MDS may be overcome by a more stringent classification according to genetic and functional data retrieved from high throughput analyses of large cohorts of patients with chronic myeloid neoplasms. Disclosure: Wolf-Karsten Hofmann: Advisory Role: Novartis Pharma, Celgene; Financing of Scientific Research: Novartis Pharma, Celgene Andreas Reiter: Advisory Role: Novartis Pharma; Financing of Scientific Research: Novartis Pharma

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Wissenschaftliches Symposium

Biologie der chronischen lymphatischen Leukämie V741

Signaling pathways in CLL: B cell receptor and BCL2 Stilgenbauer S.1 Universität Ulm, Innere Medizin III, Ulm, Germany

Chronic lymphocytic leukemia (CLL) is the most common leukemia in the western world and affects mainly elderly patients. In clinical trials, standard treatment options were established that differ mainly based on the fitness and age of the patient. The combination of fludarabine, cyclophosphamide and the CD20 antibody rituximab (FCR) is recommended for young patients without relevant comorbidity, while bendamustine and rituximab (BR) should be favored for elderly (ca. >65 years) individuals. Patients with relevant comorbidities should receive chlorambucil combined with anti CD20 antibody (Obinutuzumab or Ofatumumab). However, in 2014 new compounds were approved for patients with adverse genetic risk factors (17p-, TP53mut) and for relapsed/refractory CLL: both Idelalisib and Ibrutinib are orally bioavailable kinase inhibitors that block key regulators of central pathways in tumor cells. With both agents very impressive data are available with regard to tolerability and efficacy that change the treatment paradigm of CLL also in the front line setting. With Venetoclax a direct apoptosis inducer is in development that produces high remission rates combined with good tolerability in clinical trials. Combinations of the “novel” compounds Obinutuzumab, Ofatumumab, Idelalisib, Ibrutinib and Venetoclax will be studied over the next years in clinical trials to prolong remission duration and reduce side effects with the eventual aim to cure CLL. Disclosure: Stephan Stilgenbauer: Advisory Role: AbbVie, Amgen, Boehringer-Ingelheim, Celgene, Genentech, Genzyme, Gilead, GSK, Janssen, Mundipharma, Novartis, Pharmacyclics, Hoffmann La-Roche, Sanofi; Financing of Scientific Research: AbbVie, Amgen, Boehringer-Ingelheim, Celgene, Genentech, Genzyme, Gilead, GSK, Janssen, Mundipharma, Novartis, Pharmacyclics, Hoffmann La-Roche, Sanofi; Expert Testimony: AbbVie, Amgen, Boehringer-Ingelheim, Celgene, Genentech, Genzyme, Gilead, GSK, Janssen, Mundipharma, Novartis, Pharmacyclics, Hoffmann La-Roche, Sanofi

Fortbildung

Akute myeloische Leukämie V745

Management of patients with APL Platzbecker U.1, SAL und German APL Intergroup Universitätsklinikum Dresden, Medizinische Klinik und Poliklinik I, Dresden, Germany 1

Acute promyelocytic leukemia (APL) is considered a rare disease with estimated 200 to 300 newly-diagnosed cases per year in Germany. According to the FAB classification APL is designated as “M3 leukemia” and assigned to the WHO defined type of AML with recurrent cytogenetic abnormalities, “acute promyelocytic leukemia with t(15;17)(q22;q12), (PML/RARα) and variants”. In addition to age, the most important prognostic factor is the leucocyte count at diagnosis which divides patients into a high risk (> 10 Gpt / l) and a non-high-risk group (< = 10 Gpt / l). Through use of a risk-guided therapeutic approach the outcome of patients within both groups including risk of relapse has become almost comparable. Standard therapy of acute promyelocytic leukemia has long since relied on the combination of All-trans-retinoic acid (ATRA) and chemotherapy. The introduction of arsenic trioxide (ATO) in APL treatment has allowed to obtaining similarly high remission and survival rates coupled to significantly reduced myelosuppression. Recent results of the APL0406 trial by the GIMEMA-AMLSG-SAL study groups showed that the combination of ATRA and ATO is superior to standard ATRA and chemotherapy in

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front-line therapy of non-high-risk APL. Still, mainly early management of these patients remains a clinical challenge. Disclosure: Uwe Platzbecker: Advisory Role: Amgen, Celgene, Novartis, Janssen, Teva; Financing of Scientific Research: Amgen, Celgene, Novartis, Janssen, Teva; Expert Testimony: Amgen, Celgene, Novartis, Janssen, Teva V746

“Epigenetic therapy: the right timepoint, the right patient” Lübbert M.1 Med. Klinik I, Univ.-Klinik, Freiburg, Germany

DNA hypermethylation is one of the hallmarks of AML/MDS, and two azanucleoside hypomethylating agents (HMAs) have been approved by the EMA for treatment of these usually older patients: 5-azaCR (5-azacytidine, Vidaza) for higher risk MDS, and 5-azaCdR (Decitabine, DAC) as well as Aza for AML patients >65 years of age ineligible for standard chemotherapy, irrespective of the blast percentage in the bone marrow. Treatment is given in repeated courses and not stopped unless resistance/ failure sets in, and cannot be considered curative. Objective response rates are lower than with standard chemotherapy. Already at time of treatment initiation, it is recommended to discuss with the patients (provided they are eligible for allogeneic blood stem cell transplantation) this curative option as subsequent treatment. Thus HMAs have evolved as a frequently used treatment for “bridging” to allografting. In older, non-fit AML patients, HMAs have emerged as frontline treatment even when adverse cytogenetics are present (including patients with monosomal karyotypes which remarkably can also respond, albeit not as long as karyotypically normal patients). In initially hyperleukocytotic AML patients, the indication for an HMA, and differences in activity between the available HMAs in this not infrequent setting is as yet not firmly established (in part due to different inclusion criteria in the pivotal studies as regards WBC). Since HMAs are slow-acting drugs, initial cytoreduction of high WBCs e.g. by hydroxyurea is recommeded prior to initiation of HMA treatment. The role of HMAs as frontline treatment in fit AML patients who until now are treated aggressively is subject to clinical investigation. Specifically, the EORTC/GIMEMA/GMDS-SG are conducting an Intergroup trial 1301, comparing standard induction/consolidation (by chemotherapy or allografting) with 10-day DAC followed by allografting or continued DAC treatment (“inDACtion vs. induction”) in AML patients 60 years and older. The role of HMAs in AML relapse, particularly with “smouldering” kinetics, is not well established but may also have an important role as pre-treatment prior to allografting. In conclusion, HMA treatment of older patients can often provide well-tolerated disease control, and can be very useful as bridging treatment prior to allografting. Disclosure: Michael Lübbert: Financing of Scientific Research: Janssen-Cilag V747

Indications for allogeneic stem cell transplantation in patients with AML Passweg J.1 Universitätsspital Basel, Hämatologie, Basel, Switzerland

In 2016 the utility of allogeneic stem cell transplantation (HSCT) is clearer than in the past as several studies have shown superior results of allogeneic HSCT compared to other treatment approaches in high risk AML in 1st complete remission. Similarly it is clear that low risk AML is not benefitting from early allogeneic HSCT and these patients may undergo allogeneic HSCT after relapse although data for these indications are less solid. The situation in intermediate risk AML is more controversial and no uniform generally accepted criteria do exist. The risk stratification of AML based on genetic and mutational data has been progressively refined and this has lead to disease subgroups being moved to higher or lower risk status. E.g. FLT3 may harbour ITD mu-

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tation at a high allelic or a low allelic ratio and kinase domain mutations and it is only the high allelic ratio diseases that are considered for early allogeneic HSCT. Minimal residual disease measurement (MRD) is gaining importance in the field of AML HSCT in a similar manner to ALL. The MRD field is progressing rapidly and is complex as different types of technologies exist including cytometry, allelic burden measurement using different technologies. Some studies have shown AML with measurable MRD to be prognostically similar to chemorefractory AML and these patients would be generally selected for early HSCT. Finally there is considerable development in the field of allogeneic HSCT where protocols have been developed to accept donors other than HLA matched siblings or well matched unrelated donors. First results of unrelated donor transplantation have improved and are considered to equivalent to sibling donor HSCT and the use of haploidentical donors has increased over the last few years. Last selection of patients for HSCT has become more complex as upper age limits have been progressively removed and assessment of co-morbidities is instrumental to guide patients to the most appropriate treatment options. Therefore many issues remain: Which patients to transplant upfront, which patients to transplant after relapse, which patients not to transplant at all, who should be the best donor and this in the light of minimizing transplant toxicity and maximizing exploiting beneficial graft versus leukemia effects. Disclosure: No conflict of interest disclosed.

Fortbildung

Klug entscheiden V749

“Do” recommendations of the DGHO “Choosing Wisely” Task Group Krause S.W.1 Universitätsklinikum, Medizinische Klinik 5, Erlangen, Germany

Inspired by the “choosing wisely” campaign of medical societies in the USA, the DGHO joined a similar program of the German society of Internal medicine (DGIM). A task group within the DGHO was commissioned to select 5 topics of over-supply und 5 topics of shortage in hematology and oncology in Germany. Five recommendations to avoid unnecessary procedures were adapted and modified from ASH and ASCO and are reported separately. Recommendations for procedures that are perceived not to be performed to a sufficient degree in German practice were derived from a survey among DGHO members. The following suggestions were selected to be included in the top 5 “do” recommendations: 1) Patients with neoplastic diseases in a non-curative therapy setting should be offered dedicated palliative care. Contact to a palliative care team should be provided in time, where appropriate in parallel with tumor-specific therapy. 2) The need of psychooncological support should be evaluated in every patient with a neoplastic disease. In case of need, such support should be provided. 3) Tumor pain should be treated according to the WHO pain relief ladder. This includes pain history, individual titration of analgetics, on demand medication for breakthrough pain and treatment of side effects of opioids. 4) Possible benefits and risks have to be explained to the patient in a comprehensible way. The treatment strategy has to be negotiated considering the patients individual preferences. 5) Molecular diagnostics have to be performed in all cases in which the results may lead to meaningful implications for treatment. Interestingly, it turned out that the majority of suggestions coming up in this survey were directed toward a plus in “talking medicine” and individual patient care and not asking for a plus in high tech medicine.

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Disclosure: Stefan Krause: Advisory Role: MSD; Financing of Scientific Research: Takeda, MSD, Novartis; Expert Testimony: Affimed; Other Financial Relationships: Reiseunterstützung: Gilead, Amgen, Alexion

Wissenschaftliches Symposium Translationale Forschung V753

Prediction of immune response in hematologic and solid tumors

concern (i) the number of patients that can be offered molecular analysis in a clinical setting; (ii) the turnaround time in cases of pressing medical need; (iii) advanced bioinformatics approaches to separate driver mutations from biologically neutral passenger alterations; and (iv) the translation of molecular findings into clinical application, with limited access to drugs and a lack of molecularly stratified clinical trials being important obstacles. Furthermore, a chief priority will be exploring ways in which the use of technologies beyond genomics can help address gaps within current precision oncology initiatives to best realize the promise of informed, personalized cancer medicine. Disclosure: No conflict of interest disclosed.

Herr W.1 Universitätsklinikum Regensburg, Klinik und Poliklinik für Innere Medizin III, Regensburg, Germany 1

Spontaneous and therapeutically induced immune responses have a clear impact on the clinical course in several tumor entities. Both innate and adaptive immune cells can be effectors but also modulators of antitumor immune responses. It has been well established to analyze the expression of defined antigens (such as CD20, HER2/neu) on tumor target cells or tissues before a decision concerning the therapeutic use of a specific tumor-directed antibody is made. With the emerging availability of powerful antibodies that target immune checkpoint receptors (e.g. programmed cell death (PD)-1), more complex information is needed wether these receptors and their corresponding ligands (such as PD-L1) are expressed by immune cells and tumor cells in the tumor microenvironment. Furthermore, the impressive response rates of immune checkpoint agents in patients whose tumors carry a high mutational load constitutes a strong rationale to analyze individual tumor specimens for the presence of somatic mutations and derived neoantigens as potential predictors of treatment response. Overall this increasing interest in biomarkers and parameters that allow the prediction of immune responses in hematologic and solid tumor patients is based on major reasons. First, the new checkpoint modulating agents function systemically and can cause severe adverse reactions, making it very important to better select individual patients who will most likely benefit from treatment. Second, there is also an important ecomomic need because these modern drugs are very expensive and their tailored use including rational stratification promises to increase the cost effectiveness of treatment. Therefore, identifying relaible biomarkers for patient selection would be of great value in optimizing and personalizing tumor immunotherapy. This talk will review and discuss current biomarkers and parameters that may allow a better prediction of therapeutically induced immune responses against hematologic and solid tumors. Disclosure: Wolfgang Herr: Financing of Scientific Research: Amgen, BMS, Novartis, MSD, Celgene V754

Genomics-based individualized cancer therapy – Hype or hope? Fröhling S.1 NCT Heidelberg, Heidelberg, Germany

Precision oncology is centered on the ability to predict which patients are likely to respond to specific cancer therapies. This stratification can be achieved through different means, such as comprehensive genomic analyses of either large patient cohorts from conventional, histology-based disease categories or individual patients (often referred to as “N = 1 studies”), which can reveal alterations with immediate implications for personalized cancer therapy. Examples include actionable mutations in lung cancer, melanoma, and many other entities, which can be targeted by small-molecule kinase inhibitors, or genetic determinants of clinical benefit from immune checkpoint blockade. As an important requisite for genomics-guided oncology, next-generation DNA/RNA sequencing enables the detection of somatic alterations in cancer genomes at increasing throughput and decreasing costs. However, key areas for improvement

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Wissenschaftliches Symposium

Intensivmedizin nach allogener Stammzelltransplantation V755

HSCT and ICU – Analysis of risk factors for admission and outcome Beutel G.1, iCHOP – Intensive Care in Hematologic and Oncologic Patients Medizinische Hochschule Hannover, Hämatologie, Hämostaseologie, Onkologie und Stammzelltransplantation, Hannover, Germany 1

Background: Intensive care unit admission (ICU) may be required in stem cell transplant (HSCT) recipients who present acute illness directly or indirectly caused by transplantation procedure or complications. At this, organ involvement as respiratory failure, renal failure, or septic shock are associated with an unfavorable prognosis. While up to 35% of the HSCT recipients develop such complications and were transferred to an ICU, specific outcome parameter for HSCT recipients are underreported. Highest mortality rates were determined by the nature and number of organ failures as well as invasive mechanical ventilation. Particular in case of invasive mechanical ventilation the mortality rates range from 40% to 100%. However, recent data suggest that HSCT recipients may have a better prognosis than initially reported and mortality rates in septic patients trend close to the rate of critically patient in the general population. This observation might be an effect potentially based on the early recognition of HSCT recipients requiring intensive care. Methods: To identify risk factor for ICU admission and outcome 942 HSCT recipients transplanted between 01/2000 and 12/2013 were retrospectively analyzed. Of those 330 patients were at least once admitted to the ICU. Results: Main reasons for ICU admission were respiratory failure (n = 105, 32%), severe sepsis (n = 70, 21%), cardiac events (n = 53, 16%), and surgical procedures (n = 50, 15%). Corresponding short-term mortality was 62%, 53% and 32% for patients admitted with respiratory failure, sepsis, or cardiac events (p < 0.01). Over time the percentage of respiratory failure as a leading cause for ICU admission dropped from 40% to 27% (p < 0.01). Remarkably, overall survival after hospital discharge was comparable between ICU and non-ICU patients (52% vs. 60%, p = 0.08). Conclusion: In our cohort respiratory failure, septic shock, and cardiac events were identified as major causes of ICU admission. Long-term survival and mortality rates after hospital discharge were comparable regardless of any need for intensive care support. Comment: An overview of the recent literature and the final analysis of our data will be presented during the session. Disclosure: No conflict of interest disclosed.

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V756

Peri-procedural risk stratification in allogeneic HSCT with serial biomarkers and bed-side pulmonary function test Braun S.1, Kuhn M.2, Bergmann S.3, Alakel N.1, Schetelig J.1, Weise M.4, Kroschinsky F.1, Ehninger G.1, Höffken G.1, Bornhäuser M.1 Universitätsklinik Carl Gustav Carus, Medizinische Klinik I und Poliklinik I, Dresden, Germany, 2Institut für Medizinische Informatik und Biometrie, Dresden, Germany, 3Universitätsklinik Carl Gustav Carus, Institut für klinische Labormedizin, Dresden, Germany, 4Universitätsklinik Carl Gustav Carus, Medizinische Kliniken- Intensivstation, Dresden, Germany 1

Introduction: To evaluate the predictive utility of serial Copeptin, MRproADM, MR-proANP, NT-proBNP, IL-6, PCT, D-dimere and bed-side pulmonary function testing (PFT) for treatment failure (defined as ICU admittance or NRM) during the transplantation course in adult patients undergoing allogeneic hematopoetic stem cell transplantation (allo-HSCT). Methods: Prospective, single-center, double-blinded, thrice weekly analysis of seven biomarkers and bed-side PFT in 105 allo-HSCT patients. To avoid overestimation everyday clinical parameters, routine laboratory tests and the use of ATG were considered as confounders. Interbiomarker correlations, univariate C-statstics, best cut-offs, multivariate AIC-modelling and the continuous net reclassification improvement (cNRI) for risk prediction were calculated for the two different time windows: a) d+3 until clinical worsening and b) d +3 until ICU-transfer. Clinical worsening was defined as body temperature ≥ 38.5 °C or body weight ± 5 kg or a shock index ≥ 1. Results: High inter-biomarker correlations were found for NT-proBNP/MR-proANP (r = 0.82), IL-6/CRP (r = 0.75), MR-proADM/MRproANP (r = 0.74), MR-proADM/Copeptin (r = 0.7), MR-proADM/ IL-6 (r = 0.64). In the time frame d+3 to clinical worsening the AUCs of univariate C-statistics for IL-6 (86.69), MR-proADM (86.17) and Copeptin (82.02) were much higher compared to CRP (77.31), WBC (75.62) or creatinine (62.50). Of the seven investigated biomarkers the multivariate AIC-model selected MR-proADM (p < 0.01) and D-Dimer (p < 0.1) for risk prediction. cNRI compared to the best model of the standard biomarkers (CRP/platelets/GGT) plus patient’s age revealed high reclassification numbers for Copeptin (0,898), D-Dimers (0,918) and NT-proBNP (0,516). Despite high AUC-values the serial PFTs were not tolerated by several patients, resulting in high drop-out rates. Conclusion: Serial analysis of MR-proADM, Copeptin, D-Dimer and NTpro-BNP seem to have a predictive valueI in the peri-procedural course of adult patients undergoing allo-HSCT. These biomarkers open the door for very-early risk prediction before clinical worsening. The additional predictive value of IL-6 is limited if CRP is already increased, while Copeptin, D-Dimer and NT-proBNP still provide clinical relevant information. The WBC correlated hardly with any of the most predictive biomarkers. Disclosure: No conflict of interest disclosed. V758

Graft-versus-host-disease and Intensive Care Medicine Liebregts T.1 Universitätsklinikum Essen, Westdeutsches Tumorzentrum, Klinik für Knochenmarktransplantation, Essen, Germany 1

For many years the mortality rate of patients with hematologic malignancies requiring intensive care unit admission exceeded 90%. In recent years overall prognosis of critically ill patients with malignancies has improved with mortality rates below 50%. However Hematopoietic stem cell transplantation (HSCT) recipients are still in particular of risk for dying in ICU. HSCT is associated with multiple infectious and non-infectious complications that often lead to organ failures. Allogeneic HSCT is frequently complicated by graft-versus-host disease (GVHD) in more than half of the patients. GVHD requires reinforcement of immunosuppression resulting in sustained immunodeficiency. Pulmonary complications are particularly frequent and up to 30% of HSCT recipients require

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mechanical ventilation for acute respiratory failure. Multi organ failure, uncontrolled GVHD and invasive mechanical ventilation are associated with a poor prognosis. Although response to corticosteroids and remission status of acute GVHD at ICU admission are considered key factors, so far these factors are insufficiently assessed in present studies. Furthermore data on chronic GVHD and ICU outcome are lacking. Thus there is an urgent need to further elucidate the role of GVHD in critically ill HSCT recipients. Denying ICU access does not appear to be reasonable and broad ICU admission and extensive unlimited intensive care support is justified for patients admitted early in ICU with one isolated organ dysfunction regardless of disease and transplant characteristics. It may also be speculated that specialized HSCT-ICU may improve outcome although whether transplantation units providing critical care management attain superior outcomes need to be elucidated. Disclosure: No conflict of interest disclosed.

Freier Vortrag

AYA – Heranwachsende und junge Erwachsene V759

Germline analysis for tumor susceptibility – the next generation must go beyond organ borders: Why hereditary breast cancer is a challenge not only for gynecologists Decker J.1,2, Wohlleber E.1, Spittau G.1, Knoll N.1, Brauner M.1, Lengfeld T.1, Ritthaler M.1, Hartmann B.1, Borozdin W.1, Kohlhase J.1 Humangenetik Freiburg, Freiburg, Germany, 2III. Medizinische Klinik, Universitätsklinikum Mainz, Mainz, Germany 1

Introduction: Data of a single institution’s experience over a period of 2 years will be presented. More than 700 families met the S3 guidelines for hereditary breast and ovarian cancer (HBOC). Methods: Index patients were analyzed for BRCA1, BRCA2, and other relevant genes. In case of clear clinical indications, other genes were analyzed in addition – though not part of the canonical breast cancer genes. The pathological findings, and their clinical implications will be shown and be discussed with respect to the usefulness of established guidelines and the need for adaptive changes of these rules – beyond organ borders. Results: Family A: 2 not affected sisters were referred for BRCA testing as their sister died from breast cancer at age of 48. In accordance to the German S3 guidelines, their aunt (sister of the not affected father) who had also fallen ill with breast cancer, was tested as index patient and found to be negative for all relevant genes. The mother of the 2 not affected sisters died from pancreatic cancer at age 74, the age of onset was 74. Interestingly, there were more cases of pancreatic cancers in the lineage of the mother. Despite of age of onset being over 50 years, the molecular genetic testing for hereditary pancreatic cancer was performed in a sequential approach, including more than 10 genes. A mutation was found in the CDKN2a gene. As one of the sisters (33 years of age) tested also positive for this mutation, a thorough clinical work-up revealed an amelanotic melanoma which could be removed in sano. Family B: A woman was referred for BRCA testing as she fell ill with breast cancer at age 56, and the father’s mother had also suffered from breast cancer, however at age 88. In accordance to the guidelines, no testing for BRCA was performed. However, as the histology of the breast cancer was lobular and the patient’s father had suffered from a gastric cancer of the diffuse subtype at age 48, molecular genetic testing for CDH1 was performed and detected a clearly pathogenic mutation. Prophylactic gastrectomy was performed and five incidental cancers of a diffuse subtype were detected. Conclusion: These families and other data of our presentation clearly demonstrate that it is highly advisable to test beyond the guidelines for HBOC. This approach is lifesaving and should be considered – beyond

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the scope of gynecological tumors – particularly in the new era of next generation sequencing. Disclosure: No conflict of interest disclosed. V760

“Rather one more chemo than one less...” Young adult’s patients and chemotherapy in advanced cancer. A study of oncologists’ views Laryionava K.1, Mehlis K.1, Hiddemann W.2, Heußner P.2, Winkler E.C.1 Nationales Centrum für Tumorerkrankungen (NCT), Heidelberg, Germany, Department of Internal Medicine III, University Hospital Grosshadern; LudwigMaximilians University, Munich, Germany 1 2

Introduction: Empirical research demonstrates that there is an increasing tendency to administer cancer-specific therapy to patients with advanced cancer close to the patient’s death. The aim of this qualitative study was to understand the oncologists’ decisions to administer therapy to young adult patients at the End of Life (EoL) and to investigate the extent to which young age was a factor in cancer treatment decisions. Methods: We conducted 29 face-to face qualitative interviews with physicians and nurses working at the Department of Hematology and Oncology at the LMU University hospital in Munich, Germany. The interviews were analyzed according to the grounded theory approach. The normative implications of the results were assessed against the background of ethical principles and the concept of a fair share of life. Results: Asked about the most challenging situations in oncology, physicians and nurses frequently reported decisions and discussion about limiting cancer treatment with young adult patients and their families. “Young age” was often used as a justification for trying further aggressive treatments at the EoL. We found out that decisions were not only influenced by medical reasons and patient preferences for further therapy, but also by oncologist’s identification with patient (countertransference) and by physicians’ desire to restore a fair share of life time to young patients (fair innings argument). Conclusions: “Struggling” against the injustice together with young patient for more life-time can have quite opposite results for the patient. This perception might run the risk to neglecting ethical aspects such as the principle of beneficence and nonmaleficence and consequently lead to overtreatment of young patients at the EoL.

ing lymphoma (31.4%) and breast cancer (26.0%). Physical fatigue was scored highest (M = 45.6; SD = 28.2), followed by emotional (M = 26.7; SD = 28.8) and cognitive fatigue (M = 19.7; SD = 22.7). Women were burdened with higher fatigue in all three subscales than men (Cohens d range .35 to .44; p < .001) with the highest differences in emotional fatigue (Mean Difference = 12.7). The older age group reported higher physical (d=.22; p < .01) and emotional fatigue (d=.19; p < .05) than the younger age group. Participants who were on sick leave at time of survey reported higher emotional fatigue (d=.22; p < .05) than those who were not. Relationship status was not found to affect CRF in AYA. Conclusion: Physical fatigue seems to be the most important dimension of CRF in AYA. Women appear to be appreciably higher burdened than men and older AYA reported higher fatigue than younger AYA. However, gender seems to be the most important sociodemographic factor. Further research should examine essential medical variables associated with CRF and point out the impact of CRF on AYAs life and cancer outcome. Disclosure: No conflict of interest disclosed. V762

Adolescent and young adults versus older adults in the setting of allogeneic stem cell transplantation – two cohorts with special needs Hilgendorf I.1, Pulewka K.2, Herzberg P.Y.3, Greinix H.4, Heußner P.5, Mumm F.5, von Harsdorf S.6, Rieger K.7, Hemmati P.8, Holler E.9, Hochhaus A.1, Wolff D.9 Universitätsklinikum Jena / Klinik für Innere Medizin III, Jena, Germany, Universitätsklinikum Jena / Institut für Psychosoziale Medizin und Psychotherapie, Jena, Germany, 3Helmut-Schmidt-Universität/ Universität der Bundeswehr Hamburg Fakultät für Geistes- und Sozialwissenschaften, Hamburg, Germany, 4LKH Universitätsklinikum Graz / Universitätsklinik für Innere Medizin, Graz, Austria, 5Klinikum der Universität München / Medizinische Klinik und Poliklinik III, München, Germany, 6Universitätsklinikum Ulm / Innere Medizin III, Ulm, Germany, 7Charité – Universitätsmedizin Berlin / Charité Centrum Tumormedizin CC14 Medizinische Klinik mit Schwerpunkt Hämatologie, Onkologie und Tumorimmunologie, Berlin, Germany, 8Charité – Universitätsmedizin Berlin (CVK), Berlin, Germany, 9Universitätsklinikum Regensburg / Klinik für Innere Medizin III, Regensburg, Germany 1 2

Introduction: Cancer patients diagnosed in adolescence and young adulthood (AYA) nowadays are conceived as a specific age cohort dealing with cancer in a stage of life characterized by development, upheavals and establishment. Although research in other age cohorts pointed out that cancer-related fatigue (CRF) is one of the most common and most burdening symptoms in cancer patients, CRF in AYA is sparsely investigated and predictors are still unknown. Methods: AYA cancer patients aged 18 to 39 years at time of diagnosis who had completed acute cancer treatment (range 0 to 48 months) were surveyed with regard to their life situation and psychosocial care. CRF had been investigated using the EORTC Quality of Life Questionnaire Fatigue Module 12 (EORTC QLQ-FA12). This module consists of three subscales (physical, emotional and cognitive fatigue). The scale ranges from 0 to 100. The CRF value and the impact of sociodemographic factors on CRF in AYA were analysed. Results: A total of 577 AYA were surveyed (73.7% female, 53.0% aged between 18 to 29 years, 47.0% aged between 30 to 39 years at time of diagnosis). The most common diagnoses are haematological cancer includ-

Background: Adolescent and Young Adults (AYA) have special psychosocial needs, but little is known about these aspects in AYA after allogeneic hematopoietic stem cell transplantation (alloHSCT) in comparison to non-AYA. We therefore reevaluated data of a previously published multicenter study1 regarding this question. Methods: Patients were enrolled between 3 and 12 months after alloHSCT or in the presence of active GVHD without time limit. The 24AM questionnaire, SF-36, Human Activity Profile (HAP) and Karnofsky Index were evaluated to address these aspects in AYA and non-AYA. Groups were compared using the Mann-Whitney U-Test. A p-value < 0.05 was considered statistically significant. Results: 205 patients (66 AYA, 139 non-AYA) were investigated. The median age of the AYA and non-AYA cohorts was 29 (range, 23-36) and 52 (range, 44-57) years, respectively. On a 1 to 7 grading Likert scale non-AYA reported to be significantly more reticent (5 (4-6) vs. 4 (3-5), p = 0.04) and traditional (5 (4-6) vs. 4 (4-5), p = 0.001). Although not differing in the median, they were also more conscientious (6 (6-7), p = 0.05), reliable (6 (6-7), p = 0.02) and tidier (2 (1-4), p = 0.05). In contrast, AYA described higher impact on physical role functioning (48+41.4 vs. 28+38.1, p = 0.001), physical functioning (68+28.5 vs. 56+28.8, p = 0.002), bodily pain (73+26.3 vs. 62+28.5, p = 0.023) and emotional role function (81+35.3 vs. 66+44.3, p = 0.027) in the SF-36. General health perception, vitality, social role functioning and mental health were comparable among both groups. On HAP scale AYA reported significantly higher maximum (78+13.3 vs. 73+14.7, p = 0.003) and adjusted activity scores (67+20.1 vs. 58+19.6, p = 0.002) compared to non-AYA, but showed similar restrictions regarding Karnofsky Index. Conclusion: To gain first insights into the necessities of AYA and nonAYA, questionnaires investigating psychosocial needs and physical activ-

Abstracts

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Disclosure: No conflict of interest disclosed. V761

Cancer-related fatigue in young adult cancer patients Nowe E.1, Friedrich M.1, Leuteritz K.1, Sender A.1, Stöbel-Richter Y.1, Geue K.1 Universitätsklinikum Leipzig, Abteilung für Medizinische Psychologie und Medizinische Soziologie, Leipzig, Germany 1

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ities may help to identify specifics and create individual treatment concepts. A prospective multicenter study is ongoing to evaluate the special needs of AYA in the setting of alloHSCT. Reference: 1 Herzberg PY et al.: Bone Marrow Transplant. 2013 Jan;48(1):129–134. Disclosure: No conflict of interest disclosed. V763

Improved nutrition in Adolescents and young adults after childhood cancer. INAYA – a proof-of-concept study Quidde J.1, von Grundherr J.1, Koch B.1, Bokemeyer C.1, Escherich G.2, Valentini L.3, Buchholz D.3, Schilling G.4, Stein A.1 Universitätsklinikum Hamburg-Eppendorf, Universitäres Cancer Center Hamburg, Hamburg, Germany, 2Universitätsklinikium Hamburg-Eppendorf, Pädiatrische Hämatologie und Onkologie, Hamburg, Germany, 3Hochschule Neubrandenburg – University of Applied Sciences, Neubrandenburg, Germany, 4 Hamburger Krebsgesellschaft, Hamburg, Germany 1

Background: Multimodality treatment improves the chance of survival but increases the risk for long-term side effects in young cancer survivors, so-called ”Adolescents and Young Adults“ (AYAs). Compared to the general population AYAs have a 5 to 15-fold increased risk of cardiovascular morbidity. Thus, improving modifiable lifestyle risk factors is of particular importance. Methods: The INAYA trial included AYAs between 18-39 years receiving an intensified individual nutrition counseling at four time points in a 3-month period based on a 3-day dietary record. At week 0 and 12 AYAs got a face-to-face counseling, at week 2 and 6 by telephone. Primary endpoint was change in nutritional behavior measured by Healthy Eating Index – European Prospective Investigation into Cancer and Nutrition (HEI-EPIC). Results: 23 AYAs (11 female, 12 male, median age 21.9 ± 4.3, median BMI: 20.4 kg/m²) after completion of cancer treatment for sarcoma (n = 2), carcinoma (n = 2), blastoma (n = 1), hodgkin lymphoma (n = 12), or leukemia (n = 6) were included. The primary endpoint was met, with an improvement of 20 points in HEI-EPIC score in 52.2% (n = 12) of AYAs. At baseline, median HEI-EPIC score was 47.0 points (range from 40.0 to 55.0 points) and a good, moderate and bad nutritional intake was seen in 4.3%, 73.9% and 21.7% of AYAs. At week 12, median HEI-EPIC improved significantly to 65.0 points (range from 55.0 to 76.0 points) (p ≤ 0.001) and a good, moderate and bad nutritional intake was seen in 47.8%, 52.2% and 0% of AYAs. No change was seen in quality of life, waist-hip ratio and blood pressure. Conclusion: Intensified nutrition counseling improves nutritional behavior of AYAs. Further studies will be required to demonstrate long-term sustainability and confirm the results in larger cohorts. Trial registration: Clinical trial identifier DRKS00009883 on DRKS Disclosure: No conflict of interest disclosed. V764

Exercise induces antioxidative capacity in the muscle of BRCA mutation carriers (BIJOU study) Otto S.1, Schumann U.1, Schulz S.1, Andreß S.1, Graf T.1, Trájer E.2, Ebner F.3, Huober J.3, Janni W.3, Zügel M.1, Steinacker J.M.1 Ulm University Medical Center, Division of Sports and Rehabilitation Medicine, Ulm, Germany, 2University of Physical Education, Division of Health Sciences and Sport Medicine, Budapest, Hungary, 3University Ulm, Department of Gynecology and Obstetrics, Ulm, Germany 1

Introduction: BRCA mutation carriers (BRCA-MCs) have an increased lifetime risk for developing cancer and therefore are interested in adjuvant approaches of tumor prevention strategies. It was shown that physical activity during adolescence results in beneficial effects in adults on tumor events and BRCA1 expression rates. ROS is increased in BRCA mutation carriers; the production of oxygen-centered free radicals can lead to al-

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tered genetic expression, disease and ageing of humans. In the ongoing BIJOU study we are examining the effects of physical exercise on DNA repair complex mechanisms necessary to mitigate DNA-damaging molecules. Methods: In this two-arm, controlled trial BRCA-MCs (m/f; 18 to 69 years) are enrolled in a 6-week supervised high-intensity training intervention group (IG) versus a supervised low-intensity control group (CG), each with a frequency of three times a week at 60 min per session. Muscle biopsies and blood samples were used to evaluate the changes in energy metabolism before and after interventions using qPCR for RNA expression levels and ELISA to test marker of interests on protein level. Secondary endpoints include gene regulation of the lipid metabolism, markers of inflammation, QoL (SF-36), Life-Orientation-Test (LOT-R), anthropometric data and physical performance. Results: Preliminary results of 14 male and female participants show a significant increase of strength performance at the rowing pull machine of 39.25% in the IG (n = 9) and an increase of 8.28% in the CG (n = 5). We observed improvements in maximal aerobic power (VO2max) by 3.2% in the IG versus decreases (- 4.4%) in the CG (n = 5). Serum inflammation marker CRP decreased significantly in the IG while it was unaffected in the CG. Serum Thiol status increased significantly from 611.27± 114.9 µmol/l to 667.89± 85.3 µmol/l in the IG while it decreased in the CG (mean ± SD = -34.58 µmol/l ± 21.53). Conclusion: Preliminary data support an upregulation of thiol as a ROS scavenger. Exercise induces increases of the serum protein thiol status, which is low in several disease patterns like cancer reflects the total DNA repair capacity of the organism by detoxifying oxygen containing free radicals. By improving the cellular metabolome in this high risk group the tumor initiation in BRCA mutation carriers might be actively suppressed and conclusively an improvement of quality of life is expected. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium

Zell-basierte Immunmodulation – Gemeinsames Symposium mit der Deutschen Gesellschaft für Transfusionsmedizin und Immunhämatologie (DGTI) V766

Extracorporeal photopheresis (ECP) to treat GvHD Hackstein H.1 Universitätsklinikum Gießen und Marburg, Klinische Immunologie und Transfusionsmedizin, Gießen, Germany 1

Extracorporeal photopheresis (ECP) is an important cell-based therapy for treatment of heterogenous immune-mediated diseases such as graft versus host disease (GVHD), cutaneous T cell lymphoma and organ allograft rejection. Despite its documented clinical efficacy major mechanistic aspects are still unknown, such as the possible induction of immunostimulatory mediators by ECP as well as the blood volume required per treatment to achieve a clinical response. Recently we have reported that ECP promotes marked release of the prototypic immunostimulatory cytokine IL-1β. ECP primes IL-1β production and activates IL-1β maturation and release in the context of caspase-1 activation in monocytes and myeloid dendritic cells. Interestingly, IL-1β maturation by ECP was fully intact in murine cells deficient for caspase-1, suggesting the predominance of an inflammasome-independent pathway for ECP-dependent IL-1β maturation. Clinically, patient analysis revealed significantly increased IL-1β production in stimulated leukapheresis concentrates and peripheral blood samples after ECP. With respect to the minimal blood volume to achieve a clinical response in GVHD therapy we have developed a mini ECP technique (mini-ECP) using only 100-200 ml of whole blood for patients with contraindications for apheresis or low body weight. Mini-ECP is well tolerated and acute and chronic GVHD resolved in a significant amount of patients.

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CONTENTS AUTHOR INDEX

The identification of IL-1β after ECP provides new insight into the immunostimulatory capacity of photopheresis. The clinical results of mini-ECP indicate that processing of low volumes of blood may be sufficient for successful GVHD-therapy at least in pediatric patients with contraindications for apheresis. Disclosure: Holger Hackstein: Advisory Role: Member Scientific Advisory Board Macopharma

Freier Vortrag

Myeloproliferative Neoplasien – Klinik, Prognose V768

Splenomegaly, elevated alkaline phosphatase and mutations in the SRSF2/ASXL1/RUNX1 gene panel are strong adverse prognostic markers in patients with systemic mastocytosis Jawhar M.1, Schwaab J.1, Hausmann D.2, Clemens J.1, Naumann N.1, Henzler T.2, Horny H.-P.3, Sotlar K.3, Schoenberg S.O.2, Cross N.C.P.4,5, Fabarius A.1, Hofmann W.-K.1, Valent P.6, Metzgeroth G.1, Reiter A.1 University Medical Centre, Department of Hematology and Oncology, Mannheim, Germany, 2University Medical Centre, Institute of Clinical Radiology and Nuclear Medicine, Mannheim, Germany, 3Ludwig-Maximilians-University, Institute of Pathology, Munich, Germany, 4Wessex Regional Genetics Laboratory, Salisbury, United Kingdom, 5Faculty of Medicine, Southampton, United Kingdom, 6Division of Hematology and Ludwig Boltzmann Cluster Oncology, Medical University, Department of Internal Medicine I, Vienna, Austria 1

Introduction: Systemic mastocytosis (SM) comprises a group of rare myeloid neoplasms characterized by accumulation of mast cells (MC) in various tissues, predominantly skin, bone marrow (BM), and visceral organs. Depending on the affected organ(s), SM-related organ damage, and involvement of other hematopoietic lineages, SM can be divided into indolent SM (ISM) and advanced SM (advSM). The KIT D816V mutation is a primary driver of disease manifestations while the presence of additional mutations, e.g. in SRSF2, ASXL, or RUNX1 (S/A/R gene panel), have a strong adverse impact on disease phenotype and prognosis. Methods: Here, we evaluated the impact of clinical and molecular characteristics on overall survival (OS) in 108 patients with ISM (n = 41) and advSM (n = 67). Organomegaly was measured by magnetic resonance imaging (MRI)-based volumetry of liver and spleen. Results: In multivariate analysis of all patients, an increased spleen volume ≥450mL (hazard ratio [HR], 5.2; 95% confidence interval [CI], [2.1-13.0]; P = 0.003) and an elevated alkaline phosphatase (AP; HR 5.0 [1.1-22.2]; P = 0.02) were associated with adverse OS. The 2-year OS was 100%, 81%, and 56%, respectively (P < 0.001), for patients with 0 (low-risk, n = 37), 1 (intermediate-risk, n = 32) or 2 (high-risk, n = 39) parameters. For advSM patients with fully available clinical and molecular data (n = 60), univariate analysis identified splenomegaly ≥1200mL, elevated AP and mutations in the S/A/R gene panel as significant adverse prognostic markers. In multivariate analysis, mutations in S/A/R (HR, 3.2 [1.1-9.6]; P = 0.01) and elevated AP (HR 2.6 [1.0-7.1]; P = 0.03) remained predictive adverse prognostic markers for OS. The 2-year OS was 79% and 53%, respectively (P = 0.0003), for patients with 0-1 (intermediate-risk, n = 37) or 2 (highrisk, n = 39) parameters (Figure). Conclusion: We conclude that splenomegaly, elevated AP and mutations in the S/A/R gene panel are independent of the WHO classification and provide the most relevant prognostic information in SM patients.

Abstracts

Fig. 1. Disclosure: No conflict of interest disclosed. V769

Clinical characteristics and treatment with tyrosine kinase inhibitors in four patients with ETV6-ABL1 positive eosinophilia-associated myeloproliferative neoplasms Ahmadova G.1, Kreil S.1, Jost P.J.2, Keller P.3, Wolfensberger N.3, Maschke A.4, Schönberger B.5, Haferlach T.6, Schwaab J.1, Naumann N.1, Jahwar M.1, Cross N.C.P.7, Fabarius A.1, Hofmann W.-K.1, Haferlach C.6, Reiter A.1, Metzgeroth G.1 III. Medizinische Klinik, Hämatologie und internistische Onkologie, Universitätsmedizin Mannheim, Mannheim, Germany, 2III. Medizinische Klinik, Klinikum rechts der Isar der Technischen Universität München, München, Germany, 3Universitätsklinik für Hämatologie und Hämatologisches Zentrallabor Inselspital Bern, Bern, Switzerland, 4Hämatologie und Onkologie, Helios Klinikum Berlin-Buch, Berlin, Germany, 5Onkologisches Zentrum, Innere Medizin und Hämatologie und Onkologie, Neuburg a.d. Donau, Germany, 6MLL Münchner Leukämielabor, München, Germany, 7Wessex Regional Genetics Laboratory, Salisbury; Faculty of Medicine, University of Southampton, Southampton, United Kingdom 1

Introduction: In BCR-ABL1 positive chronic myeloid leukemia (CML), ABL1 is well known for its exquisite sensitivity to tyrosine kinase (TK) inhibitors (TKI) such as imatinib, nilotinib or dasatinib. ETV6 is a recurrent partner gene of disparate TKs, e.g. ETV6-PDGFRB, ETV6-JAK2 or ETV6-ABL1, in eosinophilia-associated myeloproliferative neoplasms (MPN-eo). Beside MPN-eo, the ETV6-ABL1 fusion gene, as consequence of rearrangements of chromosomal bands 9q34 and 12p13, has also been reported in acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML) in overall approximately 30 patients of which 12 have been treated with TKI.

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Methods: We have identified 4 male patients (median age 39 years, range 20-61) with an ETV6-ABL1 positive MPN-eo who have been treated with dasatinib (n = 2), nilotinib (n = 1) or imatinib (n = 1). Results: All 4 patients presented in chronic phase (CP) with splenomegaly, left shifted leukocytosis (median 85,000/µL, range 39,000-197,000) and significant eosinophilia (median 6,600/µL, range 6,500-9,800). Bone marrow histology was consistent with MPN-eo in CP in all 4 patients. Cytogenetics revealed a t(9;12)(q34;p13) and an ins (12;9)(p13;q34q22) in one patient each and a complex karyotype in two cases. All three patients with more than 3 months on a TKI achieved a complete hematologic remission (CHR). Complete cytogenetic remission (CCR), as assessed by cytogenetics and Fluorescence-In-Situ-Hybridisation (FISH) analysis, was achieved in 2 of 2 evaluated patients after 5 (nilotinib) and 13 (dasatinib) months, respectively. The nilotinib-treated patient also became ETV6-ABL1 negative by nested-RT-PCR at month 5, while an additional dasatinib-treated patient achieved a complete molecular remission at month 17. All pts are currently alive after a median treatment time of 13 months (range, 3-36). Conclusion: In MPN-eo, cytogenetic analysis, FISH and/or RT-PCR is a mandatory tool for the identification of potential TK fusion genes, e.g. ETV6-ABL1 in the presence of rearrangements of chromosomal bands 9q34 and 12p13. On treatment with TKI, ETV6-ABL1 positive MPN-eo patients can achieve CHR and CCR and possibly also deep molecular remissions. Close monitoring by RT-PCR is recommended for early identification of inadequate response or resistance which has been reported in the literature.

Results: Median age was 67 years and mean palpable spleen length was 13 cm below the costal margin. At data cutoff, 63% of pts remained on treatment or completed the study as per protocol. The most common hematologic grade ≥3 AEs were anemia (34%) and thrombocytopenia (15%), which led to the discontinuation of the drug in 2.2% and 3.3% pts, respectively. Nonhematologic AEs (in ≥5% of pts) were primarily grade 1/2; grade ≥3 AEs were low overall (< 2%), except pneumonia (3.9%; n= 73, 7 discontinuations) and pyrexia (2.0%; n= 37, 6 discontinuations). At weeks 24 and 48, 57% (742/1297) and 62% (588/949) of pts achieved a ≥50% reduction from baseline in palpable spleen length. 71% of pts experienced ≥50% reduction in spleen length from baseline at any time by week 72. At weeks 24 and 48, 97% (57/59) and 92% (43/47) of evaluable pts without splenomegaly at baseline continued to have a nonpalpable spleen. Clinically meaningful improvements in symptoms were seen as early as week 4 and were maintained overtime. Overall, RUX resulted in significant improvements in symptoms in 87 pts without splenomegaly at baseline. Conclusions: Reductions in spleen length and clinically meaningful improvements in were seen in majority of the pts in the JUMP study, including pts with no palpable spleen length at baseline. Overall, safety and efficacy profile of RUX in the JUMP patient population was consistent with the 2 COMFORT studies. Disclosure: Haifa Al-Ali: Employment or Leadership Position: Angestellter Arzt; Financing of Scientific Research: Novartis; Expert Testimony: Novartis Philipp Le Coutre: Employment or Leadership Position: Angestellter Arzt; Financing of Scientific Research: Novartis, BMS, Ariad, Pfizer

Disclosure: No conflict of interest disclosed. V771 V770

Safety and efficacy of Ruxolitinib in expanded-access JUMP study: an update from a cohort of 1869 patients Al-Ali H.K. , Tavares R. , Palumbo G.A. , Palandri F. , Martino B. , Liberati A.M.6, Ullrich J.G.7, Breccia M.8, Brittain D.9, Foltz L.10, Griesshammer M.11, Raanani P.12, Gupta V.13, Giraldo P.14, Ghosh J.15, Tannir B.16, Ronco J.P.16, Vannucchi A.M.17, Le Coutre P.18 1

Universitätsklinikum Leipzig AöR, Leipzig, Germany, Universidade Federal de Goiania, Goias, Brazil, 3Division of Hematology, Ospedale Ferrarotto, University of Catani, Catania, Italy, 4University of Bologna, Bologna, Italy, 5Az. Osp. Bianchi-Melacrino’-Morelli, Reggio Calabria, Italy, 6University of Perugia, Azienda Ospedaliera S. Maria, Terni, Italy, 7Unidad de Hematología, Laboratorio Doctor Grasa, Zaragoza, Spain, 8Dipartimento di Ematologia e Biotecnologie, Università “La Sapienza”,, Rome, Italy, 9Albert Alberts Stem Transplantation Unit, Pretoria East Hospital, Pretoria, South Africa, 10St. Paul’s Hospital, University of British Columbia, Vancouver, Canada, 11Klinik für Hämatologie, Onkologie und Palliativmedizin, Mühlenkreiskliniken, Johannes Wesling Klinikum Minden, Minden, Germany, 12Rabin Medical Center, Beilinson Hospital and Sackler Faculty of Medicine, Tel Aviv University, Petah Tikva, Israel, 13Princess Margaret Cancer Centre, Toronto, Canada, 14Miguel Servet University Hospital and Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Zaragoza, Spain, 15Novartis Pharmaceuticals Corporation, East Hanover, United States, 16Novartis Pharma AG, Basel, Switzerland, 17University of Florence, Florence, Italy, 18Charité – Universitätsmedizin Berlin, Berlin, Germany 1

Introduction: Ruxolitinib (RUX) is an oral, selective inhibitor of the Janus kinases JAK1 and JAK2, indicated for the treatment of disease related splenomegaly or symptoms in adult patients with primary or secondary myelofibrosis (MF). The JUMP (JAK Inhibitor Ruxolitinib in Myelofibrosis Patients) trial is a phase 3b, expanded-access trial designed to assess the safety and efficacy of RUX in MF pts prior to the commercial availability of the drug. In this analysis, we present data from 1869 MF pts treated with RUX in the JUMP study (376 pts in Germany). Methods: MF pts (intermediate-1 with palpable spleen ≥ 5 cm from costal margin, intermediate-2, and high IPSS risk category) received starting doses of RUX based on platelet counts at baseline (5 mg BID [≥50-< 100×109/L], 15 mg BID [100-200×109/L], 20 mg BID [>200×109/L]). Primary endpoint was assessment of safety and tolerability of RUX. Other endpoints included proportion of pts with a >50% reduction in palpable spleen length and patient-reported outcomes.

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Ruxolitinib / Pomalidomide combination therapy in myelofibrosis: Current data from the MPNSG-0212 trial Stegelmann F.1, Hebart H.2, Bangerter M.3, Wolleschak D.4, Griesshammer M.5, Koschmieder S.6, Hochhaus A.7, Heidel F.7, Möhle R.8, Reiter A.9, Scheid C.10, von Bubnoff N.11, Kirschbaum R.1, Reim R.1, Sutter U.1, Vetter K.1, Döhner H.1, Schlenk R.F.1, Döhner K.1 University Hospital of Ulm, Ulm, Germany, 2Stauferklinikum Schwaebisch Gmuend, Mutlangen, Germany, 3Hematology and Oncology Practice, Augsburg, Germany, 4Otto-von-Guericke University Medical Center Magdeburg, Magdeburg, Germany, 5Department of Oncology, Klinikum Minden, Minden, Germany, 6RWTH Aachen University Hospital, Department of Haematology, Oncology, Haemostaseology, and Stem Cell Transplantation, Aachen, Germany, 7 Jena University Hospital, Jena, Germany, 8University Tübingen, Hematology and Oncology, Tübingen, Germany, 9University Medical Centre Mannheim, Mannheim, Germany, 10University of Cologne, Köln, Germany, 11University Hospital of Freiburg, Freiburg, Germany 1

Introduction: We recently reported data on 28 myelofibrosis (MF) patients (pts) treated within our phase-Ib/II MPNSG-0212 trial (NCT01644110) (Stegelmann et al., ASH 2015, abstract #826). The study evaluates the combination of ruxolitinib (RUX) plus pomalidomide (POM) in MF with anemia. Methods: Primary endpoints are response after 12 28-day treatment cycles according to IWG-MRT criteria and achievement of red blood cell transfusion independence (RBC-TI). Main inclusion criterion is MF with anemia (Hb < 10 g/dL and/or RBC transfusion dependence). Pts with platelets < 100 /nL were excluded. POM dose is 0.5 mg QD, RUX starts at 10 mg BID. The first cohort comprises 37 pts. Results: Follow-up data from 31 pts are reported. Median age was 74 years (range 49-81); 55% (17 pts) had ≥1 prior therapy; 9% (3 pts) were low/ intermediate-1, 68% (21 pts) intermediate-2, and 23% (7 pts) high-risk according to DIPSS. Mutation frequencies of JAK2, MPL, and CALR were 68%, 10%, and 19%, respectively; 58% (18 pts) were at ´high molecular risk´ defined by mutations in SRSF2 (26%), ASXL1 (22%), EHZ2 (16%), and/or IDH1/2 (3%). 468 adverse events (AE) CTCAE °1-5 were recorded. Worsening of anemia within the first 6 cycles was the most frequent AE (°2-4) in 42% (13 pts), followed by fatigue (°1-2) in 35% (11 pts); 18 AEs in 12 pts were serious (SAE °2-5); 3/18 SAE were fatal (cardiac decompensation, pneumonia, septic shock); only 2/18 SAE were considered therapy-related (hemo-

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globin °4 and neuropathy °3). Treatment interruptions or dose reductions were uncommon. Median time on treatment was 12 cycles (range 2-23); 55% (17 pts) are currently on study treatment; 45% (14 pts) stopped due to no response (n = 9), death (n = 3), or leukemic transformation (n = 2). Regarding efficacy, 19% (6 pts) experienced ´clinical improvement´ in terms of ≥2 g/dL Hb increase / RBC-TI (n = 3) or spleen reduction (n = 3). Mean Hb in the total population increased from 8.8 g/dL at baseline to 9.5 g/dL at cycle 12; 31% (9 pts) continued treatment beyond cycle 12 due to Hb increase and/ or improvement of symptoms. Conclusions: Combination of RUX and POM was safe and feasible. Taking into account that the majority of pts was pre-treated and at advanced risk, a significant number experienced clinical benefit and remained on study after cycle 12. To further improve anemia response, an increase of POM to 2 mg QD is intended for the second study cohort. Disclosure: No conflict of interest disclosed. V772

Characteristics and treatment of Polycythemia vera patients in clinical practice – a multicenter outpatient survey on 1476 individuals Jentsch-Ullrich K.1, Eberhardt J.2, Zeremski V.2, Koehler M.2, Wolleschak D.2, Heidel F.H.3,4 Praxis für Hämatologie und internistische Onkologie, Magdeburg, Germany, Universitätsklinikum Magdeburg, Hämatologie und Onkologie, Magdeburg, Germany, 3Universitätsklinikum Jena, Hämatologie und Onkologie, Jena, Germany, 4Leibniz-Institut on Aging Research (FLI), Jena, Germany 1 2

Purpose: Treatment recommendations for patients with polycythemia vera (PV) are well established. While most multicenter trials investigating novel therapeutic strategies for PV are conducted at specialized academic centers, the majority of patients is treated in an outpatient setting. The aim of this study was to evaluate the ‘real live’ situation in a cohort of 1476 patients by analyzing data from a survey conducted at private practices and primary care centers. Methods: Eligible private practices and primary care centers treating patients with MPN were recruited to participate in a paper-pencil based survey conducted from March 2015 until January 2016 in Germany. Hematologists were asked to report from patient charts. Descriptive analyses were conducted to assess for outcomes and examined by reported prognostic risk scores, symptom scores, and clinical response criteria. Results: In total 34 centers participated in our retrospective survey and provided data on 1476 patients. Most patients were of older age (66.7% older than 66 years of age), which was the main risk factor. Molecular status was lacking in 23% of patients. Low rates of constitutional symptoms were reported in this physician-based survey with concentration problems, fatigue and itching being the main PV-related symptoms. Phlebotomy and hydroxyurea were the main cytoreductive measures for hematocrit control. The majority of patients were responsive (67.8%) and tolerant (77.3%) to hydroxyurea (HU) therapy. Interferon and JAK-inhibitor therapy were used in less than 10% of patients, respectively. Overall, leukocytosis, thrombocytosis and hematocrit could be effectively controlled by any therapy applied. Lack of efficacy was reported on reduction of constitutional symptoms and splenomegaly. Conclusions: The population investigated was older than participants in large multicenter trials. Age was the main risk factor accounting for the majority of patients being categorized as ‘high risk’. The physician-based survey reported on significantly lower rates of constitutional symptoms than patient-based surveys in the literature. Consistent with previously published reports, HU is the main agent used for PV therapy in an outpatient setting resulting in efficient control of hematopoietic parameters. Constitutional symptoms and splenomegaly, however, may not be reduced as efficiently sufficiently, which may be more efficiently addressed by the use of Jak-inhibitor treatment for high-risk patients.

Abstracts

Disclosure: Kathleen Jentsch-Ullrich: Financing of Scientific Research: Vortragshonorar Novartis Florian Heidel: Advisory Role: Advisory Board Novartis; Financing of Scientific Research: Vortragshonorar Novartis V773

Sequential intensified RIC versus standard RIC conditioning followed by allogeneic SCT for high risk myelofibrosis patients Christopeit M.1, Zabelina T.1, Alchalby H.1, Zeck G.1, Ayuk F.A.1, Wolschke C.1, Kröger N.1 Universitätsklinikum Eppendorf, Klinik für Stammzelltransplantation, Hamburg, Germany 1

Introduction: Patients suffering DIPSS high risk myelofibrosis are at significant risk of relapse and death. In order to improve the results of Busulfan-Fludarabine-based conditioning, we compared outcomes after conditioning with sequential FLAMSA-chemotherapy followed by Busulfan-Fludarabine-based conditioning in 17 patients with DIPSS “high risk” to the outcomes of 21 historical patients who received RIC Busulfan-Fludarabine with DIPSS “high risk”. Patients and methods: Median age of 11 female/ 27 male patients was 64 (37-75) years. Myelofibrosis was post-PV/ -ET myelofibrosis in 13 patients (34%). Conditioning consisted of iv Busulfan (10×0.8mg/kgBW) and Fludarabine (6×30mg/m²) for 21 patients. The remaining 17 patients were conditioned with Fludarabine 30mg/m², Amsacrine 100mg/m², Cytarabine 2000mg/m² on 4 consecutive days, briefly “FLAMSA”, followed by iv Busulfan (10×0.8mg/kgBW) and Fludarabine (additional 2×30mg/m²). Both regimens were combined with ATG at 30-90 mg/kg. All patients received PBSC with a median of 6(3.1-14)x106 CD34+/kgBW. A matched related donor was chosen for 5 patients (13%), 20 patients (53%) were transplanted from a matched unrelated donor, and 13 patients (34%) from a mismatched unrelated donor. Immunosuppression mainly consisted of cyclosporine and mycophenolic acid. Results: Leucocyte engraftment was reached after a median of 12 (9-32) days (FLAMSA-Busulfan), and 14 (10-30) days (Busulfan-Fludarabine, p = 0.163). Platelet recovery occurred after 20 (9-118) days (FLAMSA-Busulfan), and 22 (10-293) days (Busulfan-Fludarabine, p = 0.560). Acute GVHD grades 2-4 was experienced by 6 patients (FLAMSA-Busulfan) and 10 patients, respectively (Busulfan-Fludarabine, p = 0.444). Eleven patients in the FLAMSA-Busulfan group versus 8 patients in the Busulfan-Fludarabine group experienced chronic GVHD to some extent (p = 0.103). Cumulative incidence (CI) of relapse at 3 years was 18 ± 10% (FLAMSA-Busulfan) and 22 ± 10% (Busulfan-Fludarabine, p = 0.958). CI of non-relapse mortality (NRM) at 1 year was 26 ± 12% (FLAMSA-Busulfan) and 27 ± 11% (Busulfan-Fludarabine, p = 0.726). Kaplan-Meier estimated OS at 3 years was 63 ± 11% (Busulfan-Fludarabine) and 62 ± 12% (FLAMSA-Busulfan, p = 0.863). Conclusion: In this series of patients treated in one institution, OS, CI of relapse and CI of NRM of patients with myelofibrosis after FLAMSA-Busulfan-based conditioning was comparable to corresponding outcomes following Busulfan-Fludarabine-based conditioning. Disclosure: No conflict of interest disclosed.

Fortbildung

Publish or perish? Meet the editor V774

What is the (p-) value of the p-value? Gale R.P.1 Imperial College of London, Hematology Research Centre, Division of Experimental Medicine, London, United Kingdom 1

Inferential processes including notions of cause-and-effect are increasingly dominated by statistical analyses, a process in which the concept

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of the p-value has become central. Use of p-values in scientific reports is increasing. About 16 percent of articles in MEDLINE in 2014 mentioned a p-value including about 35 percent of meta-analyses, 40 percent of clinical trials and about 55 percent of randomized clinical trials. Most reported p-values reported cluster around p = 0.05 or p = 0.01. Interestingly, statisticians are less certain what precisely a p-value is, means and implies about reproducibility of scientific data. Some researchers think the p-value indicates the probability the null hypothesis is true or that a high p-value indicates there is no association between a variable and an outcome. Both notions are incorrect. Worries over uses and abuses of p < 0.05 as the basis for rejecting the null hypothesis has been described as having more flaws than Facebook’s privacy policy J. Some journals have gone so far as to ban use of p-values. A p-value as the probability under a specified statistical model a statistical summary of the data would be equal or more extreme than the observed value(s). When we calculate a p-value we are not testing whether the difference between groups or cohorts occurred by chance but rather consistency of the data with a proposed statistical model. Another consideration is the p-value does not reflect effect size. Clinically unimportant effects can have very low p-values when the sample size is large whereas clinically more important effects can have higher p-values because of small sample size. Estimated clinically-important effects with confidence intervals/bands and p-values should be transparently reported. It is inadvisable to focus on an arbitrary point such as p < 0.05 to justify scientific claims. Considering p = 0.05 as a point for deciding on statistical significance is arbitrary and without a sensible mathematical rationale. Other factors need consideration in considering whether an outcome is statistically significant such as study-design, measurement accuracy, evidence external to the study, accuracy of measurements and validity of assumptions underlying the data analyses. These and other issues related to using p-values in scientific publications will be discussed. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Myelodysplastisches Syndrom – Prognose, Therapie, Monitoring V779

Cytogenetic monitoring from peripheral blood in patients with myelodysplastic syndrome: First results from the German LEMON-5 trial Braulke F.1, Germing U.2, Schuler E.2, Schulz X.3, Platzbecker U.4, Nolte F.5, Hofmann W.-K.5, Giagounidis A.6, Götze K.7, Lübbert M.8, Schlenk R.F.9, Schanz J.10, Bacher U.10, Shirneshan K.10, Ganser A.11, Letsch A.12, Schafhausen P.13, Bug G.14, Brümmendorf T.H.15, Haas R.2, Trümper L.10, Haase D.10 Universitätsmedizin Göttingen, Hämatologie und medizinische Onkologie, Göttingen, Germany, 2Universität Düsseldorf, Hämatologie / Onkologie, Düsseldorf, Germany, 3Universitätsmedizin Göttingen, Medizinische Statistik, Göttingen, Germany, 4Universität Dresden, Hämatologie / Onkologie, Dresden, Germany, 5Universitätsklinik Mannheim, Hämatologie / Onkologie, Mannheim, Germany, 6Marienhospital Düsseldorf, Hämatologie / Onkologie, Düsseldorf, Germany, 7Technische Universität München, Hämatologie / Onkologie, München, Germany, 8Universitätsklinikum Freiburg, Hämatologie / Onkologie, Freiburg, Germany, 9Universitätsklinikum Ulm, Hämatologie / Onkologie, Ulm, Germany, 10Universitätsmedizin Göttingen, Hämatologie / Onkologie, Göttingen, Germany, 11Medizinische Hochschule Hannover, Hämatologie / Onkologie, Hannover, Germany, 12Charité Universitätsmedizin Berlin, Hämatologie / Onkologie, Berlin, Germany, 13Universitätsklinikum HamburgEppendorf, Hämatologie / Onkologie Hubertus Wald Cancer Center, Hamburg, Germany, 14Universitätsmedizin Frankfurt, Hämatologie / Onkologie, Frankfurt/ Main, Germany, 15Rheinisch-Westfälische Technische Hochschule Aachen, Hämatologie / Onkologie, Aachen, Germany 1

Introduction: Transfusion-dependent patients with myelodysplastic syndrome (MDS), less than 5% bone marrow (BM) blasts and an isolated deletion of the long arm of chromosome 5 (del(5q)) can be treated with

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the immunomodulatory drug lenalidomide. Within the German clinical phase II trial LEMON-5 (EudraCT-Nr.: 2008-001866-10) of the University of Duesseldorf, Germany, low- and intermediate-1-risk transfusion-dependent MDS patients with isolated del(5q) received lenalidomide and were followed by sequential cytogenetic analyses from bone marrow (BM) and peripheral blood (PB). Methods: Informations about transfusion history, BM cytomorphology, histopathology and conventional chromosome banding analyses (CBA) were documented. Additionally, patients were monitored by sequential Fluorescence-in-situ-Hybridization (FISH) analyses from immunomagnetically enriched circulating CD34+ myeloid progenitor cells (CD34+FISH) every 2-3 months from PB using comprehensive probe panels of up to 13 probes to detect the most common chromosomal aberrations typical for MDS. According to international consensus criteria the cut-off values for each probe were evaluated in our lab. Results: In 144 patients screened at study entry cytogenetic analyses were performed, 89 patients received study medication with lenalidomide (Schuler et al., 2015). Twenty percent of patients could not be enrolled in the trial because of further aberrations beside del(5q) detectable by CBA, CD34+PB-FISH or both. Hematologic response developed very fast, followed by FISH-response and CBA- or morphologic response. Cytogenetic response was detected in 74% by CD34+FISH and in 64% by CBA. Nearly all patients with cytogenetic response achieved transfusion independence (TI), but only half of the cytogenetic non-responders reached TI. Overall and AML-free survival were significantly better in cytogenetic responders than in non-responders. Conclusions: For the first time a cytogenetic monitoring from peripheral blood was performed in a prospective setting in lenalidomide treated low-risk MDS patients. Our data show that the results of CD34+PB-FISH correlate well with CBA results of BM blood, have prognostic relevance and can be used for better treatment control. Disclosure: Friederike Braulke: Financing of Scientific Research: Novartis, Celgene; Expert Testimony: Diese Studie wurde von Celgene unterstützt. Detlef Haase: Financing of Scientific Research: Novartis, Celgene; Expert Testimony: Diese Studie wurde von Celgene unterstützt. V780

Molecular characterization of therapy-related MDS after curative intensive therapy for de novo acute myeloid leukemia Böttner A.1, Herold S.1, Helas S.1, Sayehli C.2, Dührsen U.3, Hindahl H.4, Kullmer J.5, Sauer M.1, Mohr B.1, Bornhäuser M.1, Ehninger G.1, Röllig C.1, Thiede C.1, Platzbecker U.1 Universitätsklinikum Carl Gustav Carus Dresden, Medizinische Klinik und Poliklinik I, Dresden, Germany, 2Universitätsklinikum Würzburg, Zentrum für Innere Medizin, Würzburg, Germany, 3Universitätsklinikum Essen, Klinik für Hämatologie, Essen, Germany, 4St.-Johannes-Hospital, Dortmund, Germany, 5 DIAKO Bremen, Medizinische Klinik II, Bremen, Germany 1

Introduction: Secondary therapy-associated myelodysplastic syndromes (tMDS) are a very common complication following intensive chemotherapy in solid tumors and they have also been described in patients with previous acute promyelocytic leukemia (APL) several years after curative treatment. It is currently unknown whether tMDS also evolves after treatment in patients with other subtypes of acute myeloid leukemia (AML). Methods: We have identified 15 patients developing tMDS after curative therapy for de novo non-APL AML. All patients were initially diagnosed with AML and were mostly treated with conventional double induction chemotherapy using standard anthracycline/cytarabine combinations (according to the protocols of the Study Alliance Leukemia); five patients received allogeneic SCT. Patients had a median age of 56 years (range 3574) at the time of AML diagnosis and developed tMDS at a median of 46 months (range 13-132) after diagnosis. Molecular alterations in AML and tMDS counterparts were evaluated in five patients by NGS-based resequencing of 54 genes related to myeloid neoplasms (TruSight Myeloid Panel, Illumina).

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Results: Only two patients had an aberrant karyotype at diagnosis of their AML. The respective karyotypes at the time of diagnosis of tMDS are shown in Table 1 with 7 patients displaying a chromosome 7 aberration. Interestingly, in all 8 NPM1+ AML patients the mutation was not detectable at the time of tMDS diagnosis. Resequencing revealed that in two patients none of the mutations found at AML diagnosis (FLT3, NPM1; FLT3, WT1 respectively) were present at tMDS diagnosis. In two patients a typical DNMT3A mutation was present both at the time of AML and tMDS, in one patient no mutations were detected in the AML and tMDS sample. Of note, common recurrent mutations in genes such as SF3B1, ASXL1, SRSF2 and RUNX1 were not found in any of the tMDS cases. Conclusions: In summary, we describe here a series of tMDS occurring after treatment of de novo AML. The different mutational profiles of both diseases underline the existence of such a clinical entity. More studies are needed to further understand the molecular mechanisms behind this phenomenon.

chromosomal banding analysis (CBA), interphase fluorescence in situ hybridization (FISH) including a 17p13/TP53-covering probe, multicolor FISH (mFISH), sequencing of TP53 and molecular karyotyping (MK) by SNP-array analysis. The number of CA, the number of cytogenetic fusions as shown by mFISH and the size of total genomic aberrations (TGA) measured by MK quantified the extent of genetic imbalances. Results: In 58/120 (48.3%) pts. we found molecular TP53 mutations. FISH/CBA revealed deletions of TP53/17p in 27/87 pts. (31.0%). Molecular and/or cytogenetic changes of TP53 could be observed in 47/87 pts. (54.0%). In detail, a total of 29/87 pts. (33.3%) showed one alteration (chromosomal or molecular). Two TP53 aberrations were found in 18/87 pts (20.7%). Comparing pts. with and without molecular TP53 mutations we detected a higher number of CA (median 7 (range 3-41); vs 4 (3-21); p = 0.005), a larger TGA size (341 Mb (97-511); vs 105 Mb (64-335); p = 0.006) and a higher number of fusions (2 (0-9) vs 4 (0-13); p = 0.02). Likewise the number of CA and fusions was significantly higher in pts. with one or two TP53 aberrations in comparison to pts. with no TP53 aberration (CA: 8 (3-41); 8 (3-25); 4 (3-13); p = 0.00002; fusions: 5 (0-13); 3.5 (0-10); 2 (0-9); p = 0.004). Preliminary follow-up analysis suggests an additional impact of the number of CA on survival outcomes of complex aberrant pts. with molecular TP53 mutations. Conclusion: A high proportion of patients (54.0%) with complex aberrant MDS/sAML show molecular and/or cytogenetic aberrations of TP53. They show higher levels of genetic instability compared to pts. without any TP53 alteration. The extent of cytogenetic complexity may be prognostically relevant for complex aberrant MDS/AML pts. with molecular TP53 mutations. We will collect further data to prove this and complete the missing analysis. Disclosure: No conflict of interest disclosed. V782

Identification of mutations in MDS patients using next generation sequencing – Comparison of peripheral blood, CD34+ blood cells and bone marrow aspirate Martin R.1, Ganster C.1, Dierks S.1, Haase D.1 University Medical Center Göttingen, Clinics of Haematology and Medical Oncology, Göttingen, Germany 1

Fig. 1. Patient characteristic. Disclosure: No conflict of interest disclosed. V781

Interdependency between TP53 mutational status, genetic instability and prognosis in MDS and secondary AML Schaab R.1, Ganster C.1, Dierks S.1, Tallo Parra M.1, Martin R.1, Germing U.2, Platzbecker U.3, Shirneshan K.1, Lange F.4, Kröger N.5, Döhner K.6, Glaß B.7, Bacher U.1, Haase D.1 Universitätsmedizin Göttingen, Klinik für Hämatologie und Medizinische Onkologie, Göttingen, Germany, 2Universitätsklinikum Düsseldorf, Klinik für Hämatlogie, Onkologie und Klinische Immunologie, Düsseldorf, Germany, 3 Universitätsklinikum Carl Gustav Carus der TU Dresden, Medizinische Klinik I, Dresden, Germany, 4Hufeland Klinikum, Mühlhausen, Germany, 5 Universitätsklinikum Hamburg Eppendorf, Klinik für Stammzelltransplantation, Hamburg, Germany, 6Universitätsklinikum Ulm, Klinik für Innere Medizin III, Ulm, Germany, 7Asklepios Klinik St. Georg, Klinik für Hämatologie, Onkologie und Stammzelltransplantation, Hamburg, Germany 1

Introduction: Alterations of the TP53 gene, either by molecular mutations or by cytogenetic deletions of the TP53 locus are frequent in patients (pts.) with myelodysplastic syndromes (MDS) with complex aberrant karyotypes (≥3 clonal cytogenetic aberrations (CA)). Both, TP53 alterations and increased numbers of CA are prognostically adverse in MDS. This study analyzes the interdependency between the TP53 mutational status and the extent of (cyto)genetic instability as well as the pat. prognosis. Methods: We comprehensively characterized 120 pts. with MDS (N = 100) and sAML after MDS (N = 20) with complex aberrant karyotypes by

Abstracts

Introduction: Myelodysplastic syndromes (MDS) are clonal stem cell diseases of the bone marrow (BM) characterized by a dysfunction of hematopoiesis commonly resulting in peripheral cytopenias, dysplasia and an increased risk of acute myeloid leukemia (AML) development. In up to 90% of MDS cases acquired somatic mutations account for MDS formation and constitute important markers for diagnosis. Currently BM aspirates serve as gold standard for cytogenetic and molecular genetic analysis of MDS patients. The aim of this study was to investigate weather the high sensitivity of next generation sequencing (NGS) allows reliable genetic diagnostics also from peripheral blood (PB) samples. Methods: Thirteen patients (6x RCMD, 3x RAEB-2, 2x MDS-U, 1x MDSRA, 1x CMML) were included in our pilot study. For all patients genomic DNA was extracted from mononuclear bone marrow (BMMC), peripheral blood (PBMC) and immunomagnetically enriched CD34+ peripheral blood cells (CD34+). Sequencing was performed using the TruSight Myeloid Sequencing-Panel and a MiSeq instrument (Illumina). Reads were aligned with BWA (hg19) and further processed using the software packages Picard, GATK and Annovar. T-cells were used to prove somatic origin. Results: Somatic mutations were discovered in 12 patients. Overall we found 42 aberrations in our cohort (10x TET2, 6x ASXL1, 5x SRSF2, 2x U2AF1, 2x RUNX1, 1x DNMT3A, 1x TP53, 1x NRAS, 1x ETV6, 1x STAG2, 1x BCOR, 1x IKZF1) of which 32 mutations were detectable in all and 5 mutations were recovered in at least two tested sample types. Inconsistent recovery was apparent mostly for Indels. The median number of mutations found in individual patients correlated with bone marrow blast counts (≥5%: n = 3.6, < 5%: n = 2.4). Interestingly, the determined allele frequencies (AF), which correlate with the clone size of the malignant

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cells, were similar across all sample types (median AF: BMMC = 32%, PBMC = 25%, CD34+=34%; p = 0.55). Conclusion: Our findings indicate that NGS enables adequate detection and quantification of somatic mutations also from PB preparations. With exception of Indels, the concordance for the detection of a mutation between the analyzed materials was very high. The AF of individual mutations in PBMC did not significantly deviate from the AF determined in BMMC or CD34+ cells. NGS of PB would significantly reduce invasive clinical interventions and could therefore improve diagnostics and disease monitoring. Disclosure: No conflict of interest disclosed. V783

The surface antigen profile in patients (pts) with Acute Myeloid Leukemia (AML) identifies subgroups characterized by distinct biological and clinical features Jentzsch M.1, Bill M.1, Schumann L.1, Leiblein S.1, Schubert K.1, Grimm J.1, Bergmann U.1, Pleß M.1, Schmalbrock L.1, Schulz J.1, Knyrim M.1, Franke G.N.1, Behre G.1, Pönisch W.1, Vucinic V.1, Niederwieser D.1, Schwind S.1 Universitätsklinik Leipzig, Hämatologie und internistische Onkologie, Leipzig, Germany 1

Despite being a long-established part of the standard diagnostic workup, the biological & clinical implications of antigen expression profiles at AML diagnosis have not been studied using a larger antigen panel. We analyzed 232 AML pts (age 61 [14-76] years) receiving myeloablative or non-myeloablative allogeneic stem cell transplantation. Mutation (mut) status of NPM1, CEBPA, IDH1, IDH2, TET2 &DNMT3A genes, presence of FLT3-ITD & -TKD & BAALC, ERG, MN1, EVI1 & GPR56 expression were assessed. Flow cytometric analysis of mononuclear bone marrow cells utilizing a standard panel was performed at diagnosis. Unsupervised hierarchical clustering revealed 4 subgroups with distinct surface antigen profiles (Fig 1).

Fig. 1. IP-Cluster.

Cluster 1 was characterized by high lymphoid, erythroid & thrombocytic differentiation antigens. Cluster 1 pts were less likely to have de novo AML (P < .01), a CBF-AML (P = 0.03) & to be TET2mut (P = 0.05) & more likely to have del5/5q (P = 0.01), del7/7q (P < .01), monosomal karyotype (KT, P < .01) & European LeukemiaNet (ELN) adv risk (P < .01) & had higher GPR56 (P < .01) expression. Cluster 2 was characterized by high monocyte & neutrophil differentiation antigens. Cluster 2 pts were more likely to have de novo AML (P < .01), to be NPM1mut (P = 0.02),

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less likely to have del7/7q (P = 0.01) & had lower BAALC (P < .01), ERG & MN1 expression (P < .01 each). Cluster 3 was characterized by the lowest CD34 & highest CD33 expression. Cluster 3 pts were more likely to have de novo AML (P = 0.02), ELN fav risk (P = 0.03), normal KT (P < .01), to be NPM1mut (P < .01), to have FLT3-ITD (P = 0.04) & had lower BAALC (P < .01), MN1 (P < .01) & GPR56 expression (P = 0.01). Cluster 4 was characterized by an immature phenotyp. Cluster 4 pts were less likely to be NPM1mut (P < .01) & had higher BAALC, ERG & MN1 expression (P < .01 each). While no outcome impact of the clusters was seen in the entire set, when only pts with a normal KT were considered, cluster 4 pts had a significantly higher cumulative incidence of relapse (P = 0.04). The surface antigen profile at diagnosis obtained by unsupervised cluster analysis associated with distinct biological & clinical features, indicating an underlying biologic connection. Disclosure: No conflict of interest disclosed. V784

Rigosertib activates canonical Wnt pathway in osteoblasts of patients with MDS but has no influence on bone turnover in healthy mice Bulycheva E.1, Rauner M.2, Weidner H.1, Hofbauer L.2,3, Platzbecker U.1,3 Universitätsklinikum Carl Gustav Carus Dresden, Medizinische Klinik und Poliklinik I, Dresden, Germany, 2Universitätsklinikum Carl Gustav Carus Dresden, Medizinische Klinik III, Dresden, Germany, 3German Cancer Consortium (DKTK), partner site Dresden and German Cancer Research Center (DKFZ), Heidelberg, Germany 1

Introduction: Rigosertib is a novel drug under investigation in clinical trials in patients with myelodysplastic syndromes (MDS). Acting as a “Ras mimetic” and affecting the PI3K and other signaling pathways, rigosertib induces cell death in MDS-derived cell lines. Healthy cells, including normal hematopoietic stem cells, are thought to be unaffected by rigosertib. Its effect on bone marrow microenvironment, particularly on osteoblasts (OB), has not been yet investigated. Methods: Mesenchymal stromal cells (MSC) were collected from MDS patients (45-78 years), differentiated towards OB and treated with 0.1µM rigosertib. Mineralization capacity was determined using Alizarin Red staining; cell viability was tested by Cell Titer Blue Assay, apoptosis – by Caspase-Glo 3/7 Assay. Alkaline phosphatase (ALP) expression was determined by RT-PCR and normalized to β-actin. Wnt signaling array from SABioscience of rigosertib-treated OB was performed. A TCF/LEF-reporter assay was done using rigosertib-treated murine C2C12 cell line. Micro-computed tomography was performed using femurs of 10-weekold male C57BL/6 mice, treated with rigosertib (250 mg/kg 3x/week i.p. for 3 weeks) or vehicle. In addition, serum was analyzed by ELISAs to assess bone turnover markers (CTX-I and P1NP). Results: Rigosertib significantly reduced viability of MDS-MSC (-8%,p  <  0.05), at least partially due to increased apoptosis (+14%,p  <  0.05). OB differentiation was affected neither in Alizarin staining, nor by ALP expression (fold increase, 1.29 vs. 1.04, p = 0.63). Rigosertib significantly reduced the expression of genes inhibiting canonical Wnt pathway (APC, FRZB, PRICKLE1). Treatment of C2C12 cell line with rigosertib also demonstrated dose-dependent activation of TCF/LEF (max. at 5 µM, 4-fold, p < 0.001). In vivo experiments showed that rigosertib did not affect bone parameter in healthy mice (trabecular bone volume 7.2% vs. 6.6%, p = 0.6; bone mineral density 119.6 vs. 105.6 mg/cm3, p = 0.45). In parallel, serum markers of bone turnover did not change significantly. We observed anemia and thrombocytopenia as side effects in mice. Conclusions: Rigosertib activates canonical Wnt pathway in MDS-OB, however, it is not accompanied by changes in cell differentiation in vitro or bone turnover in vivo. Whether modulation of the niche cells might be at least in part responsible for the therapeutic activity of rigosertib in MDS or the development of peripheral cytopenias in mice remains to be investigated.

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Disclosure: Ekaterina Bulycheva: Expert Testimony: Onconova; Other Financial Relationships: Alexion Uwe Platzbecker: Advisory Role: Celgene; Expert Testimony: Onconova

Freier Vortrag

Lymphome experimentell V785

The JAK inhibitor Ruxolitinib offers an effective treatment strategy for peripheral T-cell lymphomas Proske A.1, Kissel S.1, Zwick A.1, von Bubnoff N.1, Duyster J.1, Dierks C.1 Universitätsklinik Freiburg, Klinik für Innere Medizin I, Freiburg, Germany

Introduction: Peripheral T-cell lymphomas (PTCLs) carry a dismal prognosis with high relapse rates and high mortality due to the lack of specific therapies and resistance to classical chemotherapies. About 20% of PTCLs carry the ITK-SYK fusion oncogene, which mimics constitutive T-cell receptor signaling. ITK-SYK overexpression in mice induces a peripheral T-cell lymphoma with malignant T-cells infiltrating bone marrow, spleen, skin and other organs, mimicking the human disease. In addition the malignant T-cells induce an inflammatory phenotype with upregulation of pro-inflammatory cytokines as well as the recruitment of granulocytes into various organs. Furthermore, we find a strong activation of JAK/STAT signaling in the malignant cells. Results: Ruxolitinib is a kinase inhibitor targeting JAK1, JAK2 and TYK2 and is broadly used for the treatment of myeloproliferative neoplasms. Due to hyperactivation of JAK/STAT signaling in our mouse model, we aimed to investigate the effects of Ruxolitinib in T-cell lymphomas. In vitro treatment of human and murine T-cell lymphomas showed reduced proliferation and enhanced apoptosis. In vivo experiments were performed and mice carrying the ITK-SYK oncogene were treated with Ruxolitinib for 5 weeks. While all control mice developed lethal T-cell lymphomas within 60 days after transplantation with massive weight loss, visible skin infiltrates and splenomegaly, all Ruxolitinib treated mice survived with only mild signs of disease and stable weight during the whole treatment period. Organ analysis revealed that Ruxolitinib treatment normalized the T-cell distribution and differentiation of the ITK-SYK+ malignant T-cells within thymus and bone marrow. In addition, we found a strong reduction of inflammatory cytokines in Ruxolitinib treated mice, and granulocytosis in the peripheral blood as well as granulocytic infiltrates in different organs were strongly reduced. In contrast to Ruxolitinib, Pacritinib which targets JAK2 and TYK2, had only minor effects on disease development, indicating JAK1 as the major mediator of malignant transformation. In addition, only Ruxolitinib, but not Pacritinib was able to normalize STAT3 and STAT5 phosphorylation levels in malignant T-cells. Conclusion: Our results demonstrate the requirement of an intact JAK/ STAT signaling pathway for T-cell lymphoma development and implicate Ruxolitinib as a highly effective treatment strategy for peripheral T-cell lymphomas. Disclosure: No conflict of interest disclosed.

V786

Genomic profiling of T-cell Prolymphocytic Leukemia (T-PLL) identifies novel recurrent mutations in SAMHD1 and PTPRC Johansson P.1,2, Klein-Hitpass L.2, Bergmann A.3, Siebert R.3, Scholtysik R.2, Przekopowitz M.2, Seifert M.2, Zenz T.4,5, Dührsen U.1, Küppers R.2,5, Dürig J.1,5 Klinik für Hämatologie, Universitätsklinikum Essen, Universität DuisburgEssen, Essen, Germany, 2Institut für Zellbiologie (Tumorforschung), Medizinische Fakultät, Universität Duisburg-Essen, Essen, Germany, 3Institut für Humangenetik, Christian-Albrechts-Universität Kiel und Universitätsklinikum Schleswig-Holstein, Kiel, Germany, 4Abteilung Translationale Onkologie, Nationales Zentrum für Krebserkrankungen, Deutsches Krebsforschungszentrum (DKFZ), Heidelberg, Germany, 5Deutsches Konsortium für Translationale Krebsforschung (DKTK), Heidelberg, Germany 1

Introduction: T-PLL is an aggressive leukemia, which derives from mature T-lymphocytes. It is characterized by resistance to chemotherapy and associated with a poor prognosis. Disease defining genetic alterations resulting in TRD/TRA-TCL1A or TRD/TRA-MTCP1 fusion proteins are not sufficient for the development of the disease, additional genetic lesions like mutations or copy-number variations are needed. Inactivating mutations or deletions of ATM and gain-of-function mutations of JAK1/3 and STAT5B are recurrently involved in T-PLL pathogenesis. PTPRC, encoding the CD45 molecule and involved in the JAK/STAT pathway, is mutated in CD45-negative T-ALL. SAMHD1 has been described as a tumor suppressor gene, which is mutated in about 11% of CLL patients. Methods: We isolated CD3/CD4 positive tumor cells from peripheral blood samples of T-PLL patients by sort purification and performed single nucleotide polymorphism (SNP) microarray analysis, next-generation RNA sequencing and targeted capture sequencing. CD3-positive cells from five healthy donors enriched by magnetic cell separation were included as controls. SNP analysis and RNA sequencing were performed for 11 patients and five healthy donors. Validation of sequencing data was performed on DNA level for 51 candidate genes using a capture approach (Roche NimbleGen seqcap choice) in a cohort of 31 patients. RNA and targeted sequencing were performed on Illumina HiSeq 2500. Results: SNP chip analysis in 11 patients revealed both gains and losses (incl. uniparental disomies) on various chromosomes. We confirmed previously described copy number variations (CNVs) on chromosomes 8, 11, and 22. Additionally, we observed recurrent CNVs on chromosomes 1q and 20q (regions for PTPRC and SAMHD1) in 25% and 17% of patients, respectively. RNA sequencing results, confirmed by targeted capture sequencing, revealed mutations for PTPRC and SAMHD1 in further 13% and 16% of patients, respectively. Conclusions: Genetic profiling of T-PLL identified novel CNVs and mutations in PTPRC and SAMHD1, revealing additional mechanisms in the molecular pathogenesis of T-PLL. Disclosure: No conflict of interest disclosed. V787

Nuclear Interaction Partner of ALK (NIPA) is involved in NPMALK driven lymphomagensis Lippert L.J.1, Shlyakhto V.1, Kreutmair S.1, Klingeberg C.1, Albers C.2, Miething C.1, Duyster J.1,3, Illert A.L.1,3 Uniklinik Freiburg, Klinik für Innere Medizin I, Hämatologie, Onkologie und Stammzelltransplantation, Freiburg, Germany, 2III. Medizinische Klinik, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany, 3Deutsches Konsortium für Translationale Krebsforschung (DKTK) und Deutsches Krebsforschungszentrum (DKFZ), Heidelberg, Germany 1

Anaplastic large cell lymphoma (ALCL) is a high grade lymphoma with T- or null cell phenotype. Translocation t(2;5)(p23;q35) in 60% of ALCL leads to expression of the oncogenic tyrosine kinase NPM-ALK. NIPA is an F-Box-Protein contributing to the timing of mitotic entry by defining an oscillating ubiquitin E3 ligase. Co-expressed with NPM-ALK, NIPA is constitutively phosphorylated and functionally inactivated. However, the

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functional consequences of this interaction as well as the role of NIPA in NPM-ALK induced lymphomagenesis remains unknown. In this study, we identified NIPA as a crucial protein for effective ALCL induction and lymphoma progression. Softagar-Assays of primary Nipa/MEFs showed a significant decrease in colony formation upon NPMALK infection (38 vs. 79 CFUs; p < 0.001). Targeted genetic approaches substantiated these results with prolonged transformation and reduced proliferation of NPM-ALK infected Ba/F3 upon NIPA downregulation (74% of wt growth; p < 0.001). Infection of Karpas299 with NIPA miR led to a significant reduction of the proliferation ability compared to ctrl miR cells (66% of wt growth; p < 0.01). Interestingly, treatment with the ALK inhibitor TAE-684 suggested synergistic effects of ALK inhibition and NIPA knockdown. To investigate the impact of NIPA deletion in vivo, we used a retroviral bone marrow (BM) TX model resembling human ALCL. Based on a Cre/loxP system under the LCK-Promotor, NPM-ALK expression and NIPA-deletion are restricted to early T cells. Within this model, wt mice develop Thy1.2+ lymphoma with a latency of 4-6 months and die from neoplastic T cell infiltration of bone marrow and lymphatic organs. First results from mice transplanted with Lck-CreTG/wtNIPAflox/floxBM show a trend to impaired disease induction and progression. Secondary transplantation of CD4-/CD8-lymphomas led to a distinct deceleration of disease development upon NIPA deletion. Moreover, a long-latency model of NPM-ALK expression in enriched HSCs induced a significantly prolonged survival (110 vs. 80 days; p < 0.01) and reduction of spleen colonies (10 vs. 28 colonies/spleen; p < 0.001) in mice transplanted with MigNPM-ALKNIPAko/ ko BM compared to control animals, suggesting a crucial role of NIPA in NPM-ALK driven lymphomagenesis. Taken together, our data highlight the importance of the NIPA/NPMALK axis in ALCL development and progression, and may thereby elucidate a novel molecular target for therapeutic intervention. Disclosure: No conflict of interest disclosed. V788

The TYK2-STAT1 pathway in aggressive T-Cell Lymphoma: A new therapeutic intervention site? Prutsch N.1, Sanda T.2, Wu L.3, Turner S.4, Jaeger U.5, Staber P.5, Steinberger P.6, Moriggl R.7, Mueller M.7, Kenner L.1, Merkel O.1 Medizinische Universität Wien, Institut für experimentelle Pathologie, Wien, Austria, 2Cancer Science Institute, Singapore, Singapore, 3Department of Immunology, Genentech Inc, San Francisco, United States, 4Division of Molecular Histopathology, Department of Pathology, University of Cambridge, Cambridge, United Kingdom, 5Medizinische Universität Wien, Department of Medicine I, Clinical Division of Hematology and Hemostaseology and the Comprehensive Cancer Center, Wien, Austria, 6Medizinische Universität Wien, Institut für Immunologie, Wien, Austria, 7University of Veterinary Medicine, Institute of Animal Breeding and Genetics, Wien, Austria 1

Whole genome sequencing of T-cell lymphomas has revealed mutations in Januskinase family genes (JAK1, JAK2, JAK3, TYK2), known to have essential functions in cytokine signalling, motility and proliferation. Targeting Januskinases has opened new therapeutic intervention sites in cancer and autoimmune diseases. Tyrosinekinase 2 (TYK2) the first JAK kinase to be identified has important roles in inflammation and anti tumor immunity. Here we report pathway dependence on TYK2 in anaplastic large cell lymphoma (ALCL), a CD30 positive, aggressive Non-Hodgkin T-cell lymphoma. The importance of TYK2 is underlined by the recent description of TYK2-fusions driving oncogenic signalling in ALCL. Interestingly, using published RNAseq data of 23 primary ALCL cases, TYK2 expression was generally high as compared to PBMC independent of the presence of TYK2-fusions, suggesting active TYK2 signalling in large fraction of patients. Ablating TYK2 in ALCL cell lines, using lentivirally transduced shRNAs as well as CRISPR/CAS9 technology, led to rapid cell death induction. To test the therapeutic relevance of our findings, ALCL cell lines and PBMCs were treated with small molecule inhibitors of TYK2 (TYK2#2,Bayer-18) leading to apoptosis induction exclusively in tumor cells. Immunoblotting and shRNA mediated knockdown of potential

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downstream targets of TYK2 revealed an essential function for STAT1 in ALCL cell survival. Flow cytometric analyses revealed active regulation of PD-L1 in TYK2 depleted cells. Tumor intrinsic and immune-ceckpoint relevant consequences of TYK2 knockout are currently being assessed using an ALCL mouse model with T-cell specific TYK2 knock-out and experiments employing T-cell activation reporter cell lines with and without PD-1 expression. Disclosure: No conflict of interest disclosed. V789

Pharmacological restoration and therapeutic targeting of the B-cell phenotype in classical Hodgkin’s lymphoma Neuendorff N.R.1,2, Du J.2,3, Yu Y.4, von Kries J.P.5, Mathas S.1,4, Dörken B.1,4, Lee S.2,4, Schmitt C.A.1,2,4 Charité Universitätsmedizin Berlin, Klinik für Hämatologie, Onkologie und Tumorimmunologie, Campus Virchow Klinikum, Berlin, Germany, 2Molekulares Krebsforschungszentrum (MKFZ), Charité Universitätsmedizin Berlin, Berlin, Germany, 3Center of Cancer Research, Binzhou Medical University Hospital, Binzhou, China, 4Max-Delbrück-Center for Molecular Medicine, Berlin, Germany, 5 Leibniz-Institut für Molekulare Pharmakologie, Screening Unit, Berlin, Germany 1

Introduction: Classical Hodgkin’s lymphoma (cHL), although originating from B-cells, is characterized by the virtual lack of gene products whose expression constitutes the B-cell phenotype. Epigenetic repression of B-cell-specific genes via promoter hypermethylation and histone deacetylation as well as compromised expression of B-cell-committed transcription factors were previously reported to contribute to the lost B-cell phenotype in cHL. Restoring the B-cell phenotype may not only correct a central malignant property, but render cHL susceptible to clinically established antibody therapies targeting B-cell surface receptors as well as small compounds interfering with the B-cell receptor (BCR) signaling. Methods: We conducted a high-throughput pharmacological screening based on more than 28,000 compounds in cHL cell lines carrying a CD19 reporter to identify drugs that promote re-expression of the B-cell phenotype, and exploited the co-therapeutic potential of B-cell-targeting antibodies and small compounds. Results: Our high-throughput pharmacological screening retrieved three chemicals that robustly enhanced CD19 transcription. Subsequent chromatin immunoprecipitation (ChIP)-based analyses indicated that two of these compounds exerted their action by lowering transcriptionally repressive lysine 9-trimethylated histone H3 (H3K9me3) levels at the CD19 promoter. Furthermore, the anti-leukemia agents all-trans retinoic acid (ATRA) and arsenic trioxide (ATO) were found to reconstitute the silenced B-cell transcriptional program and reduce viability of cHL cell lines. When applied in combination with a screening-identified chemical, ATO evoked re-expression of the CD20 antigen, which could be further therapeutically exploited by enabling CD20 antibody-mediated apoptosis of Hodgkin cells. Moreover, restoration of the B-cell phenotype also rendered cHL cells susceptible to the B-cell Non-Hodgkin’s lymphoma (B-NHL)-tailored small compound inhibitors Ibrutinib and Idelalisib. Conclusions: Based on a high-throughput pharmacological screening, we identified different compounds that induced a re-expression of B-cell specific genes, thereby rendering cHL cells susceptible to B-cell targeting therapeutics as a conceptually novel and promising treatment strategy for this lymphoma entity. Disclosure: Nina Neuendorff: Other Financial Relationships: Stipendium der Firma Medac Clemens Schmitt: Advisory Role: Roche; Financing of Scientific Research: Roche

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V790

Proteasome inhibition and induction of cell death by EGCG [(-)-Epigallocatechin-3-gallate] in chronic lymphocytic leukaemia Ponath E.1, Schnabl S.1, Hilgarth M.1, Demirtas D.1, Reiter M.1, Hubmann R.1, Zielinski C.1, Jäger U.1, Shehata M.1 Medical University of Vienna, Vienna, Austria

Introduction: There is growing evidence that EGCG [(-)-Epigallocatechin-3-gallate] has anti-neoplastic effect in solid tumours and haematological malignancies including chronic lymphocytic leukaemia (CLL) particularly in the early stages of the disease. EGCG has a wide range of biological effects including the regulation of proteasome activity. However, no information is available on the direct effect of EGCG on the proteasome activity or its interaction with other proteasome inhibitors in CLL. Therefore, the aim of this study was to evaluate the effect of EGCG on cell viability and proteasome activity as a single agent or in combination with other proteasome inhibitors including Bortezomib, (boronic acid-based) and MG132 (non-boronic acid-based). Methods: Peripheral blood mononuclear cells (PBMC) of CLL patients were exposed to a wide range of concentrations of EGCG (1-200µM) and cell viability was evaluated by CellTiter-Blue (CTB) assays and Anx/PI staining. In parallel, the effect on proteasome activity was evaluated by Proteasome-Glo Chymotrypsin-Like Assays. Results: CTB viability assay, which reflects the cellular metabolic capacity, demonstrated a dose and time dependent decrease in the cell viability with an IC50 ranging between 50-80µM EGCG (25-50µg/ml). Apoptosis assay using Annexin V/propidium iodide staining showed a moderate increase in early (Anx+/PI-) and late apoptotic/necrotic (Anx+/PI+) after 2-3 days exposure to EGCG. Proteasome activity assay showed a remarkable inhibition of the chymotrypsin-like activity by EGCG after 4h and this inhibitory effect was more prominent after 24h. Comparative studies showed that EGCG has a moderate proteasome inhibition activity compared to Bortezomib and MG132. However, EGCG had an additive effect in combination with MG132 but an antagonistic effect against Bortezomib. Conclusion: These data demonstrate that EGCG induces cell death in leukemic CLL cells through a complex mechanism which involves the inhibition of the proteasome activity as an early event. The results substantiate the therapeutic value of EGCG and points to its potential beneficial effect in combination with non-boronic acid-based proteasome inhibitors or other drugs which target the proteasome activity in CLL. Disclosure: No conflict of interest disclosed.

Freier Vortrag Palliativmedizin V791

First results of the prospective „Palliative Care in Rostock: Focus on quality of Life (PaRoLi)” study – prevalence of anxiety and depression Sewtz C.1, Hlawa W.1, Kriesen U.1, Große-Thie C.1, Kragl B.1, Glaeser D.1,2, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany, 2Clinic of Internal Medicine III – Hematology/Oncology/Hemostaseology, Palliative Care, Complementary Medicine, Klinikum Südstadt Rostock, Rostock, Germany 1

Introduction: Health-related quality of life (HRQoL) is an important outcome measure in palliative care (PC) given that maximizing QoL is the overall goal of PC. The individuality of QoL as well as the PC setting itself makes an evaluation of HRQoL difficult. A continuous quality control is necessary, as studies have shown that the assessment by doctors differ from the patient self-assessment. The individual mood is an important part of QoL being negatively influenceable by depression and anxiety.

Abstracts

Here, we report on results of screening for anxiety and depression in a large German PC cohort. Method: From 5/2013 to 12/2013 consecutive PC patients being admitted to the interdisciplinary PC unit of Rostock University Medical Center were enrolled. QoL and mood were assessed 24h after admittance, within 24h before discharge and 2 weeks after discharge using the Hospital Anxiety and Depression Scale (HADS-D) and the Functional Assessment of Chronic Illness Therapy (FACIT-Pal). Results: Of the 206 inpatients, 102 consented to participate in the study, 104 were not included. Reasons for non-participation will be described. The study cohort consisted of 46 (45%) females and 56 (55%) males with a median age of 69 yrs (range: 44-89 yrs). Underlying diseases were in 99% of all cases oncological (hematological 15%, solid tumor 85%). Most important results were: 176 HADS-D questionnaires were completed with a mean anxiety score of 7.21 ± 3.71 and a mean depression score of 9.27 ± 4.80. Probable depression or anxiety (≥11) was present in 46% of the pts. In longitudinal analysis both subscales showed little improvement of anxiety and depression during hospitalization (p = n.s.) but significant deterioration between discharge and 2 weeks after (anxiety subscale (p = 0,046), depression subscale (p = 0,003)). Anxiety and depression correlated with each other slightly at baseline and high (r = 0.84) at home. Answers of the HADS-D questionnaire were partly influenced by patient characteristics. Further detailed data will be presented. Conclusion: QoL and satisfaction of care include many different and individually weighted aspects. They are important parameters in PC that have to be evaluated frequently. Palliative patients suffer more likely from depression than other patient groups. There is slight improvement of anxiety and depression during hospitalization but significant deterioration 2 weeks after discharge. Disclosure: No conflict of interest disclosed. V792

Assessment of psychic burden and palliative care needs of patients after first diagnosis of incurable cancer – Arbeitsgemeinschaft Palliativmedizin (APM) Project of the Section B of the German Cancer Society Vogt J.1, Sistermanns J.2, Kuon J.3, Kahl C.4, Alt-Epping B.5, Stevens S.6, Ahlborn M.7, Heider A.8, Tienken M.1, Kubin T.9, Loquai C.10, Boehm A.11, Stahlhut K.12, Oechsle K.13, Mehnert A.14, van Oorschot B.15, Thomas M.3, Ortmann O.16, Lordick F.1 Universitätsklinikum Leipzig, Universitäres Krebszentrum, Leipzig, Germany, 2Kliniken Maria Hilf Mönchengladbach, Klinik für Strahlentherapie, Mönchengladbach, Germany, 3Thoraxklinik im Universitätsklinikum Heidelberg, Internistische Onkologie der Thoraxtumoren, Translational Lung Research Center, Heidelberg, Germany, 4Klinikum Magdeburg gGmbH, Klinik für Hämatologie, Onkologie und Palliativmedizin, Magdeburg, Germany, 5 Universitätsmedizin Göttingen, Klinik für Palliativmedizin, Göttingen, Germany, 6Kliniken Essen Mitte, Klinik für Internistische Onkologie/ Hämatologie, Essen, Germany, 7Städtisches Klinikum Braunschweig, Klinik für Hämatologie und Onkologie, Braunschweig, Germany, 8Klinikum Leverkusen, Klinik für Hämatologie/ Onkologie/ Palliativmedizin/ Schmerztherapie, Leverkusen, Germany, 9Klinikum Traunstein, Abteilung für Hämatologie/ Onkologie/ Palliativmedizin, Traunstein, Germany, 10Universitätsmedizin Mainz, Hautkrebszentrum Rhein-Main, Mainz, Germany, 11Universitätsklinikum Leipzig, Klinik und Poliklinik für Hals-, Nasen-, Ohrenheilkunde, Leipzig, Germany, 12Immanuel Klinik und Poliklinik Rüdersdorf, Praxis für Hämatologie/ Onkologie/ Palliativmedizin, Rüdersdorf, Germany, 13Universitätsklinikum Hamburg-Eppendorf, Klinik und Poliklinik für Onkologie und Hämatologie, Hamburg, Germany, 14Universitätsklinikum Leipzig, Abteilung für Medizinische Psychologie und Medizinische Soziologie, Leipzig, Germany, 15Comprehensive Cancer Centre Main-Franken, Klinik und Poliklinik für Strahlentherapie, Würzburg, Germany, 16Caritas-Krankenhaus St. Josef Regensburg, Klinik für Frauenheilkunde und Geburtshilfe, Lehrstuhl der Universität, Regensburg, Germany 1

Introduction: Current data as well as medical experience suggest that palliative aspects of patient care needs should be addressed early in patients (pts) with incurable cancer. However, it has not been systematically inves-

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tigated which symptoms and specific burden exist at the time point of first diagnosis of an incurable malignant disease and how care needs change over time. Moreover, information is sparse on patients´ current needs as well as which needs are fulfilled and what demands exist for general and specialized palliative treatment although this information is required for early integration of palliative care. Methods: The APM project was initiated to assess care needs and preferences of patients suffering from incurable cancer using validated questionnaires (FACT, SEIQoL-Q, PHQ-4, modified SCNS-SF-34, Distress Thermometer) at the time of first diagnosis and before any specific anti-cancer treatment. Patients are re-assessed in the further course of disease 3, 6, and 12 months after inclusion. Medical information from the treating physicians are collected at the same time points. Results: 300 cancer patients were included into this ongoing study from October 2014 to January 2016. Median age of patients is 63 years (range 29 - 89). ECOG performance status at first visit ranges between 0 - 4. Underlying diagnoses are: lung cancer (121 pts), gastrointestinal cancer (99 pts), head and neck cancer (37 pts), gynecological cancers (35 pts) and malignant melanoma (8 pts). Preliminary results show a broad spectrum of patients´ needs and disease burden including psychological distress even at the very beginning of an incurable course of disease which will be presented in detail. Conclusion: Patients’ needs and burden including psychological distress comprise a broad spectrum even at first diagnosis of an incurable cancer. These findings emphasize the need for early individualized provision of multi-faceted support, including palliative care services with psychological support, beginning already at the time of diagnosis of incurability. Disclosure: No conflict of interest disclosed. V793

Glioblastoma multiforme in palliative care setting – life expectancy, life span and quality of life Jacobs S.1, Junghanss C.1, Große-Thie C.1, Henze L.1, Gläser H.1, Murua Escobar H.1, Kriesen U.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany 1

Introduction: Glioblastoma multiforme (GM) is an aggressive brain tumor with a poor prognosis. Due to rapid growth paterns patients often suffer from symptoms caused by intracranial pressure and localized disturbances. As cognitive function is often altered during disease course not only patients but also relatives might benefit from palliative care approaches. This study aims at characterization of glioblastoma patients who were referred to an academic palliative care ward. Method: This retrospective study included patients with glioblastoma who were hospitalized between 8/2009 and 12/2015 in the palliative care ward of the University of Rostock. Patients’ characteristics including age at time of diagnosis, reasons for as well time referral and hospitalization, advance directives, social care level, main symptoms and therapy of the glioma were analyzed. Results: The median patients age at diagnosis was 61 years (range 46-86 years). The median time between diagnosis and contact to the palliative care was 31 weeks and overall survival 37 weeks. Social decompensation (54%), central neurological disorders (57%) and psychological and mental changes (40%) were the most common reasons for referral to palliative care. 46% of the patients were initially treated with a combined therapy consisting of surgery, radiotherapy and chemotherapy while 11% received no GM specific therapy. According to the HOPE questionnaire most important physical symptoms were weakness and fatigue. Non-physical symptoms were supply problems, need for help and excessive demands on relatives. Patients were discharged to hospice (31%), their homes (family or nursing homes (40%)) or died at palliative care ward (20%). Conclusion: Glioblastoma patients and their relatives often suffer from disease associated neurological and mental symptoms causing palliative care referral. As social decompensation was also among the main reasons

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for admission early integration of PC into oncological treatment concepts seem advisable. Disclosure: No conflict of interest disclosed. V794

Development of an ethics policy for advanced care planning and limiting treatment (EPAL-Study) Jaeger E.1, Mehlis K.2, Mumm F.1, Laryionava K.2, Hiddemann W.1, Winkler E.C.2, Heußner P.1 Klinikum der Universität München, Medizinische Klinik III, München, Germany, Nationales Centrum für Tumorerkrankungen (NCT), Heidelberg, Germany

1 2

Introduction: End-of-life (EOL) decision-making is often surrounded by clinical, ethical and psychological conflicts and is therefore a challenging process for patients, relatives and their medical team. The aim of the EPAL study (Ethics policy for advanced care planning and limiting treatment) is to develop a clinical practice guideline about limiting treatment which focuses on the identification of conflicts during the process of EOL decision making, the timing of consultations, the effort of common decision-making regarding end of life as well as legal and ethical issues. Further the impact of the guideline on medical practice, patient’s information needs, patient involvement and timing of advanced care planning is evaluated in a pre-post-design. Methods: A pre-implementation study was conducted involving the medical team and the patients (with relatives) of the Department of Haematology/Oncology at the LMU Hospital (Munich). The results of this study were fed into the process of guideline development which lasted from May 2014 to July 2015 and followed the Appraisal of Guidelines for Research & Evaluation (AGREE). After the discussion of drafts by all stakeholders concerned in five different meetings, and after the review by external experts, the guideline was approved and enacted in a final clinic wide consensus conference. Results: The approved guideline does not only contribute through definitions and legal information to clarifying end-of-life situations but also provide practical support to the professionals involved through its 3 statements and 20 recommendations. With the enactment of the guideline the Department´s documentation forms had to be adapted accordingly. Simultaneously, the introduction of the guideline could be officially declared as part and parcel of the clinic´s quality management. Already during the development of the guideline (the talks, interviews, discussions) a growing awareness of the issue and an increased readiness to cooperate could be ascertained. Conclusion: The relevance of the guidelines´ concentration on the decision-making process of limiting treatment for advanced cancer patients has been underlined by a wide interest for the subject area and the guideline among other Departments. Expectantly, the post-implementation study of the guidelines´ impact, begun in January 2016, will corroborate this in a more detailed manner. Disclosure: No conflict of interest disclosed. V795

End-of-life care in patients after allogeneic stem cell transplantation Schmidt V.1, Jülich A.1, Buchhold B.2, Busemann C.1, Neumann T.1, Krüger W.H.1 Universitätsmedizin Greifswald, Klinik für Innere Medizin C – Hämatologie und Onkologie, Stammzelltransplantation, Palliativmedizin, Greifswald, Germany, 2 Universitätsmedizin Greifswald, Institut für Medizinische Psychologie, Greifswald, Germany 1

Introduction: Allogeneic stem cell transplantation (alloSCT) is a curative approach for a variety of hematological diseases with an otherwise fatal course. Development of modern conditioning protocols and improvement of supportive therapy have made alloSCT available for elderly and pre-treated patients. Survival after alloSCT depends from factors such as

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diagnosis, biology of disease, stem cell donor, medical pre-condition of the recipient and shows a wide variation from 5% up to >80%. However, the long-term overall survival of all patients is approximately 50%. Vice versa, the other half of the patients could benefit from need palliative care at the end of their disease course. The most awesome break for the patient after alloSCT is the disruption when it becomes clear, that the curative concept of transplantation needs abruptly to be abandoned and a palliative concept is necessary for further management of disease. Patients and methods: Data from 123 patients, who had undergone alloSCT and have died after grafting, were extracted from the charts. Parameters collected were diagnoses, details about transplantation, complications such as infections, GvHD and relapse of underlying disease, and symptoms at the end of life. Additionally, measures, drugs and care at the end of life were interpreted. Results: In case of fatal course after alloSCT the majority of patients died within the 1st year (69%) and 50% within five months after transplantation. Main cause of death was relapse of underlying disease followed by infections. Bleedings, GvH-related complications, reactivation of viral infections and secondary tumours were less frequent. Main symptoms prior to death or last discharge were weakness, poor appetite, fatigue, need for aid (daily activities) and fever/infection related to pancytopenia. Majority of patients were free of pain (76%), nausea (76%), emesis (93%), dyspnea (77%), obstipation (96%), and depression (85%). Opioids, chosen as indicator drug class for admission route, were given i.v. in more than 50%. Conclusion: There is a relevant demand for high quality palliative care of patients not cured by alloSCT at their end of life. Symptoms and requirements of these patients differ from those of patients with other fatal diseases and this should be considered in patients care concepts. In addition, the abrupt change from a curative concept to a short-term infaust prognosis may raise specific psychological and spiritual requirements in these patients. Disclosure: No conflict of interest disclosed. V796

Family caregiver’s satisfaction with care within specialized inpatient palliative care – findings from a pilot study Ullrich A.1,2, Ascherfeld L.1, Marx G.3, Bokemeyer C.1, Bergelt C.2, Oechsle K.1 Universitätsklinikum Hamburg-Eppendorf, 2. Medizinische Klinik, Bereich Palliativmedizin, Hamburg, Germany, 2Universitätsklinikum HamburgEppendorf, Institut für Medizinische Psychologie, Hamburg, Germany, 3 Universitätsmedizin Göttingen, Klinik für Palliativmedizin, Göttingen, Germany 1

Introduction: Perceived dissatisfaction with care is described as a main factor for burden in family caregivers of advanced cancer patients. However, systematic literature on specific aspects determining satisfaction with specialized inpatient palliative care (SPC) is limited. This pilot study aimed to evaluate caregiver’s satisfaction with SPC to further minimize potential burden and to improve the responsiveness of SPC to caregiver’s needs. Methods: A consecutive cohort of 51 main caregivers of advanced cancer patients receiving SPC completed a questionnaire including the FAMCARE-2 (satisfaction), DT (distress), GAD-7 (anxiety), PHQ-9 (depression), FIN (needs) and socio-demographic variables. The FAMCARE-2 is a validated 17-item instrument used to measure caregiver’s perceived satisfaction with the multi-professional SPC team’s behaviors (scale: 1785, higher values reflect greater satisfaction). Four subscales are suggested: management of physical symptoms and comfort, provision of information, family support and patient psychological care. We used Spearman correlations to explore factors potentially related to the degree of care satisfaction. Results: The mean age of caregivers was 56 years (SD 15.5, range: 20-87), 56% were male, and 53% patients’ spouses or partners. With respect to the total FAMCARE-2 mean, the overall satisfaction with palliative care provision was 73.4 points (SD 8.3, range: 51-85). On average, caregivers reported high satisfaction across the subscales: 22.4 points in the physical symptoms and comfort scale (scale: 5-25), 16.5 in the information scale

Abstracts

(scale: 5-20), 17.0 in the family support scale (scale: 5-20) and 17.5 in the patient psychological care scale (scale: 5-20). We found no significant associations between the total FAMCARE-2 mean and socio-demographic factors (caregiver’s age, gender, educational level, working status, relationship to patient, prior involvement in caring for patient), psychological burden (distress, anxiety, depression), or the degree of caregiver’s needs. Conclusions: Caregiver’s were highly satisfied with the care provided within SPC, and the level of overall satisfaction was unaffected by the investigated factors. Despite of the exploratory character of the study, our findings suggest a high correspondence of the provided SPC and caregiver’s expectations, and appropriate quality of care regardless of caregiver’s characteristics. Acknowledgement: The study was funded by the Hamburg Cancer Society. Disclosure: No conflict of interest disclosed.

Debatten

Risikoadaptierte Therapie des multiplen Myeloms V800

Risk driven decision for high-dose therapy in multiple myeloma – No Goldschmidt H.1 Universitätsklinikum Heidelberg, Medizinische Klinik, Innere Medizin V, Heidelberg, Germany 1

High dose therapy of multiple myeloma (MM) is the standard of care for younger patients. There is an ongoing discussion about the age which is correlating with the term “younger MM patients”. In many MM clinical trials the patients’ upper age limit is 65 years for autologous blood stem cell transplant (ABSCT). In the GMMG study group the upper age limit for ABSCT candidates is 70 years. In single center approaches the maximum age for a MM-ABSCT candidate is most often 75 years. The results from the Italian and French randomized prospective clinical trials, comparing new drugs plus/minus ABSCT, confirmed the prolongation of PFS in the ABSCT cohort. New drugs or new drug combinations before and after ABSCT induce deeper remission. The results from the French IFM 2009 trial show that the deepness of minimal residual disease, measured with the next generation sequencing (NGS) or flow activated cell sorting, correlates significantly with prolonged progression (PFS) and overall survival (OS). The best predictor for improved PFS and OS based on the multivariate analysis used in the IMF trial was especially those MRD with negativity of greater than 1 positive cell in 1 million cells in NGS. New immune based approaches for multiple myeloma are being investigated. A specific autologous cell therapy like CAR-T Cells will further improve the MM treatment results. Disclosure: No conflict of interest disclosed. V801

Risk-adapted strategy for autologous stem cell transplantation (ASCT) in multiple myeloma (MM) - Pro Engelhardt M.1, Ihorst G.2, Waldschmidt J.1, Naegele M.1, Dold S.1, Zober A.1, Kiote-Schmidt C.1, Bertz H.1, Einsele H.3, Goldschmidt H.4, Straka C.5, Duyster J.1, Wäsch R.1 University of Freiburg Medical Center, Hematology, Oncology & Stem Cell transplantation; Comprehensive Cancer Center Freiburg (CCCF), Freiburg, Germany, 2Clinical Trials Center, University of Freiburg, Freiburg, Germany, 3 Department of Internal Medicine II, CCC Mainfranken, University Hospital Würzburg, Würzburg, Germany, 4Hematology, Oncology & Rheumatology, National Center for Tumor Diseases, Heidelberg University Hospital, Heidelberg, Germany, 5Schön Klinik Starnberger See, Starnberg/München, Germany 1

Introduction: Evidence-based medicine and reliable data from well conducted randomized clinical trials (RCT) support the routine use of high-

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dose therapy (HDT) and ASCT in eligible patients (pts) ≤70 years (yrs) of age. Data on ASCT have been generated before and after the introduction of novel agents (NAs), the latter initially non-RCTs leading some physicians to believe that a risk-adapted strategy should be routinely used, with biological parameters guiding treatment and ASCT being postponed to 2nd-line therapy. Methods: We performed an extensive literature review, accumulating all available data in MM, assessing current ASCT-efficacy, ongoing trials, SAEs, cost issues and future perspectives. Results: 7 randomized CTs have investigated conventional chemotherapy vs. ASCT; all of which were in favor of ASCT in terms of response, and in 6/7 and 3/7 in EFS and OS, respectively. With NAs, induction responses increased to >70%, further upgraded with melphalan-conditioning and NA-based consolidation and maintenance, leading to PCR-/ MRD-negativity. Optimal strategies may thus result in unprecedented 5-yr OS rates of 80%. The high efficacy of NAs have led to meta-analyses and RCTs testing ASCT vs. NA-treatment alone; these meta-analyses, EMN- (NCT00551928/01208766) and IFM/US-RCTs (NCT01208662) confirming, that ASCT improves responses, PFS and OS, and that 2nd-line ASCT is possible in only ~60%. Thus, delaying ASCT carries the risk of the procedure becoming unfeasible due to refractory disease, deterioration of performance status (PS) or comorbidity. Current risk tools involve pts´ biological age, PS, organ function, cytogenetics, GEP, plasma cell labeling index, R-ISS and others; albeit risk-adapted strategies for or against ASCT are rarely tested, rather than the latter considered via valid assessment tools, testing ASCT eligibility. World-wide performed RCTs challenge best NA-based induction, consolidation and maintenance vs. ASCT (GMMG, DSMM), in elderly pts (DSMM XIII), best comorbidity/ risk-tools for fitness/ASCT-eligibility and pt-related health programs with special nursing and training support to alleviate ASCT symptoms. Efficacy calculations show that ASCT remains a cost effective procedure in 90% of pts, bears low mortality rates, and remains the standard of care, with NA-based induction and consolidation. Conclusion: In pts ≤70yrs, tested fit for ASCT, HDT/ASCT remains the standard of care associated with prolongation of PFS and OS. Disclosure: Monika Engelhardt: Advisory Role: Janssen, Celgene, MSD, Amgen, Novartis; Financing of Scientific Research: Janssen, Celgene, MSD, Amgen, Novartis; Expert Testimony: Janssen, Celgene, MSD, Amgen, Novartis Ralph Wäsch: Advisory Role: Celgene, MSD, Amgen, Novartis; Financing of Scientific Research: Celgene, MSD, Amgen, Novartis

Wissenschaftliches Symposium

Biologie des myelodysplastischen Syndroms

Disclosure: No conflict of interest disclosed. V803

CHIP, ICUS, IDUS et al.: New diseases or just new classifications? Heuser M.1 Medizinische Hochschule Hannover, Hannover, Germany

The increased use of molecular diagnostics in hematology often reveals gene mutations, while clinical or morphologic signs of a hematologic malignancy are missing. In cytopenic patients one is likely to diagnose myelodysplastic syndrome (MDS). However, to prevent unnecessary overdiagnosis of MDS a new entity has been defined, which may precede a myeloid malignancy as monoclonal gammopathy of undetermined significance (MGUS) does for multiple myeloma. Clonal hematopoiesis of indeterminate potential (CHIP) represents a new disease entity, which requires a somatic mutation in blood or bone marrow, but does not fulfill other criteria of known hematologic malignancies. The prevalence of CHIP increases with age and is found in 10% of healthy people in their seventies. We estimate that 2.75 million people are affected in Germany. The most frequently mutated genes are DNMT3A, TET2 and ASXL1. The transformation rate to a hematologic malignancy is 13-fold increased and reaches 0.5-1% per year. CHIP should be considered in the differential diagnosis of patients with cytopenias and will contribute to a better understanding of the clinical significance of clonal hematopoiesis. Disclosure: No conflict of interest disclosed. V804

V802

Molecular pathophysiology in diagnostics and treatment of myelodysplastic syndromes

Novel targets and innovative therapeutic agents in MDS Wermke M.1 Uniklinikum Dresden, Medizinische Klinik I / UCC Early Clinical Trial Unit, Dresden, Germany 1

Schneider R.K.1 Klinik für Hämatologie, Onkologie, Hämostaseologie und SZT, Med. Fakultät Uniklinik RWTH Aachen, Aachen, Germany 1

Myelodysplastic syndromes (MDS) are often challenging to diagnose for several reasons, e.g. the wide range of clinical presentations and the quantification of morphologic features. A more objective mechanism for diagnosing MDS, in particular in confounding cases, would be of great clinical benefit. The clinical heterogeneity of MDS reflects the diversity of molecular abnormalities that drive disease pathogenesis. Technological developments have enabled the identification of many new genetic lesions in MDS patients, providing profound insights into disease pathogenesis. However, even with these advances, our understanding of how these mutations contribute to the etiology, pathogenesis, and therapeutic responses of MDS remains largely unknown. Improved animal models provide greater insights into the disease biology and will hopefully accelerate the development of new therapeutic agents.

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Using somatic mutations to diagnose MDS is complicated by several issues: (1) the absence of mutations in most cases, (2) mutated genes are not highly specific for MDS and (3) clonal hematopoiesis is common in older individuals and we lack outcome data for patients with clonal cytopenias of undetermined significance (CCUS). The full impact of genetics on MDS, however, has not yet been fully realized. As with the del(5q) chromosomal abnormality some gene mutations may be strongly associated with clinical features and thus be used to help classify MDS subtypes in patients that meet the classical diagnostic criteria. Molecular factors may soon be integrated into diagnostic criteria and clinical management algorithms, as specific mutations are shown to distinguish disease subtypes and provide prognostic information not captured by current risk assessment models. In the future, clinical variables, pathologic features, and genetic information will be integrated for prospective risk stratification and the choice of specific therapies for the treatment of MDS.

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With only three licensed drugs and almost a decade without the approval of a novel therapeutic agent, there is still a high unmet clinical need in MDS. Amelioration of disease-inherent cytopenias remains the main therapeutic target in lower risk MDS and may be achieved by erythropoietin and its derivatives in a subset of patients. It has become increasingly clear that GDF11 controlled late stage erythropoiesis represents another attractive therapeutic target in MDS as well and Luspatercept, a selective GDF11 antagonist, has demonstrated remarkable erythropoietic responses especially in MDS with ring sideroblasts. The development of thrombopoiesis stimulating agents (TSA) has been more challenging due to initial concerns regarding a pro-leukemic potential of these drugs. However, mature data on romiplostim and recent results regarding eltrombopag demonstrated safety and efficacy of TSA in lower risk MDS Hypomethylating agents (HMA) are the mainstay of higher risk MDS treatment, although responses are usually not durable. Therefore, novel

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approaches to either increase HMA efficacy and/or to tackle HMA-resistance are actively pursued. Clinical trials assessing HMA in combination with several histone-deacetylase inhibitors have, however, largely failed to substantially improve outcome. Novel, and potentially more efficient strategies include the combination of HMA with inhibitor of aberrant RAS-signaling and inducers of apoptosis. Reactivation of anti-tumor immune-activity by targeting PD1/PD-L1 or CTLA4 immune checkpoints might be another way to overcome HMA resistance, which is currently tested in clinical trials. Another, more direct way to re-induce anti-tumor immune activity are bispecific T-cell engaging antibodies and receptor-modified T-cells as well as vaccination strategies, which are actively developed for late-stage MDS and AML patients. In conclusion, although therapeutic advances in MDS are coming slowly therapeutic progress is evident and may change our treatment algorithms in the near future. Disclosure: No conflict of interest disclosed. V805

Diagnostic and prognostic parameters in therapy-related MDS Kuendgen A.1 Heinrich-Heine University Düsseldorf, Department of Hematology, Oncology and Clinical Immunology, Düsseldorf, Germany 1

Therapy-related myeloid neoplasms (t-MN) can occur in susceptible individuals as a late effect of either cytotoxic or radiation treatment of a primary malignant or autoimmune disease. Until the last WHO classification all t-MN were lumped together independent whether they represented t-MDS, t-MPD, or t-AML. The only differences made were due to the causative compound (alkylating agent vs topoisomerase inhibitor). This categorization was due to early publications describing a frequency of clonal abnormalities >90%, including mostly high-risk cytogenetic subtypes with a predominance of chromosome 5 and 7 abnormalities. T-MN appeared to have a uniformly poor prognosis. It is now widely recognized that t-MDS and t-AML, although having a lot in common, are separate entities and should be treated as such. Prognostic research with a focus on t-MDS, however, is scarce. Many colleagues apply the IPSS or IPSSR to patients with t-MDS, although both scoring systems have been developed excluding such patients. The only scoring system specific for t-MDS has recently been published by Quintas-Cardama et al. Since further international data is needed, our Austrian-German-Swiss study group (DACH) together with the Spanish MDS group and other European as well as US centers supported by the IWG for MDS (International Working Group) have now put together a database including 1837 pts with t-MDS diagnosed 1975-2015. Complete data to calculate the IPSS/-R was available in 1511 patients. In a first step the impact of prognostic features was analyzed by uni- and multivariable models and estimated by a measure of concordance for censored data. By these methods we could show that IPSS and IPSSR, can be applied in t-MDS. However, both scoring systems perform inferior when compared to their performance in p-MDS. This opens the way for further analyses based on our dataset aiming at the establishment of a t-MDS specific score, since it is obvious from existing data that treatment decisions must be individualized and not all t-MDS patients are poor-risk and will benefit from intensive treatment approaches. In addition to conventional prognostic data, more and more molecular data will be collected and improve our knowledge on t-MDS development and prognosis. Currently TP53 mutations appear to be the most frequent molecular subtypes, existing at least in a subgroup of patients already before treatment of the primary disease and representing one possible new mechanism of t-MN development. Disclosure: Andrea Kuendgen: Financing of Scientific Research: Vorträge für Celgene und Novartis; Expert Testimony: Celgene

Abstracts

Wissenschaftliches Symposium Biologie des Multiplen Myeloms V806

The genetic machinery of multiple myeloma Driessen C.1 Kantonsspital St Gallen, St.Gallen, Switzerland

A better understanding of the genetic basis of multiple myeloma, its variability, cloncal and subclonal evolution and its link to MM cell biology is likely to strongly influence a rational approach to MM therapy. Key areas to address include the clonal heterogeneity of MM in individual patients as well as between them, the cloncal and sub-clonal evolution of such changes and to what extent this is shaped by therapy and/or different biological MM subtypes, and ultimately how such changes are related to drug sensitivity, resistance and clinical outcome of MM patients. Recent technological advantages allow to dissect the genetic machinery of maligant diseases, including multiple myeloma (MM), in considerable detail. They currently demonstrate for MM the presence of on average 2-7 clones with 20- > 100 subclonal variants in individual patients, so that 20-80% of MM cells carry subclonal genetic variations. In particular increased activities in the RAS, MYC and NFkB pathways have been identified as MM drivers. While there is no single dominant mutation found in MM, > 75% of MM cells carry mutations in a small number of hot spot genes including KRAS, NRAS, FAM46C, DIS3, TP53, BRAF, TRAF3, CYCLD or RB1. This cloncal and subclonal heterogeneity is present already at the M-GUS stage. The clonal evolution of MM during the lifetime of the disease may follow stable, alternating, linear or branched patterns, which may have implications for the choice of therapy. To directly connect genetic changes to functional biology and therapeutic options, we shall as an example dissect the genetics and functional biology of proteasome inhibitor resistant MM. It shall be demonstrated how a complex pattern of well-tailored changes in gene and protein expression in MM cells undergoing proteasome inhibitor treatment drives a highly specialized network of functional changes in maturation status, drug export proteins and metabolic homeostasis. This results in drug resistance and ultimately clonal advantage and selection, and at the same time offers novel therapeutic targets specifically addressing the biology of this newly selected clone. Disclosure: Christoph Driessen: Advisory Role: Celgene, Amgen; Financing of Scientific Research: Celgene, Amgen, Bristol-Meyers Squibb, Novartis, Takeda, Mundipharma; Expert Testimony: Amgen, Novartis, Celgene, Mundipharma V807

Minimal residual disease (MRD) investigation in multiple myeloma – hype or real progress? Hose D.1, Seckinger A.1 Medizinische Klinik V, Universitätsklinikum Heidelberg, Heidelberg, Germany

Introduction: Assessment of minimal residual disease (MRD) aims at detection of low numbers of myeloma cells remaining in the bone marrow after successful treatment, with sensitivity above “conventional” methods (e.g. immunofixation). Methods: Three main methods are used: i) “Next generation” flow cytometry, typically with 8-colour panels, e.g. the EuroFlow (two tubes, (1) CD45, CD38, CD138, CD19, CD56, B2M, cyIgκ, cyIgλ (detection of aberrant plasma cells), (2) CD45, CD38, CD138, CD19, CD27, CD28, CD117, CD81 (further phenotypic characterization)), ii) allele-specific oligo nucleotide polymerase chain reaction (ASO-PCR) detecting rearranged and thus myeloma cell specific hypervariable heavy- or light chain regions, and iii) next generation sequencing (NGS), detecting myeloma cell specific DNA-alterations. For PCR and NGS-based methods, the malignant phenotype must, for flow cytometry should be characterized up-front. Results: Sensitivity varies from 1:5×105 - 1:106 (flow cytometry), to 1:106 (ASO-PCR and NGS), with a theoretical limit given by the number of

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cells aspirated (typically 2-5×107) and Poisson-distribution. The feasibility varies from 95% of patients in case of flow cytometry (e.g. GMMG-MM5 trial) to >90% for NGS and ASO-PCR, if heavy and light chains are investigated using up-front CD138 purification, in clinical routine and multicenter trials. MRD only assesses the response level at the site of bone marrow aspiration and thus needs to be complemented with whole body imaging techniques (e.g. whole body MRI). MRD-negativity thus does not mean absence of remaining myeloma cells. The potential clinical use of MRD is twofold: i) prognostication (deeper remission is associated with longer progression free and overall survival), and ii) guiding treatment, in terms of a) stopping (aggressive) treatment in MRD-negativity, or b) intensifying treatment to reach MRD-negativity. Conclusion: The ability to detect myeloma cells after successful treatment at a level of 1:106 or beyond in clinical routine or multicenter trials in almost all patients is a real progress. It should be considered standard for response assessment and evolves as powerful tool for guiding treatment within clinical trials. One of the first such trials will be the GMMG-CONCEPT-trial starting in fall 2016. Outside clinical trials or scientific research programs, there is today no indication for MRD-diagnostics in multiple myeloma. Disclosure: Dirk Hose: Advisory Role: Celgene; Expert Testimony: Celgene, Jansen, Novartis, EngMab, Sanofi Anja Seckinger: Expert Testimony: Celgene, Jansen, Novartis, EngMab, Sanofi V808

Immunotherapy in Multiple Myeloma Knop S.1 Med. Klinik und Poliklinik II, Würzburg, Germany

Most likely, the first systematic investigation into immunotherapy of multiple myeloma (MM) was performed with the administration of interferon alpha (IFN). A meta analysis, published in 2001, reported on more than 4.000 myeloma subjects who had been treated with IFN during induction and maintenance. A marginal increase in event-free and overall survival was documented for those studies. However, adverse effects along with the introduction of high-dose chemotherapy and the initial compounds of the two novel classes of drugs (the proteasome inhibitors [PIs] and the immunomodulatory compounds [IMiDs]) led to the abrogation of IFN use in MM. The widely used IMiD, lenalidomide, is known for its anti-myeloma effects. However, more recently its immunomodulatory properties have been evaluated. Lenalidomide was shown to induce formation of immunological synapses (via cortical actin); to enhance NK cell response; and to expand active T lymphocyte subsets. This makes the compound particularly attractive as an enhancer of different immunotherapeutic approaches in MM. Besides allogeneic stem cell transplant (allo SCT), use of cellular immunotherapy (adoptive T cell transfer and chimeric antigen receptor-modified T cells [CARTs]) and myeloma-targeting monoclonal antibodies (moAbs) are of particular interest. Despite the introduction of next-generation PIs and IMiDs, myeloma patients continue to relapse, rendering allo SCT (followed by lenalidomide in some instances) an attractive option. The evaluation of CARTs is ongoing in multiple hematologic cancers due to their outstanding efficacy. Those autologous, genetically modified T cells express the antigen-binding domain from a B cell receptor fused to the signaling elements associated with a T cell receptor. Thereby, the engineered T cell is redirected to its antigenic target and is activated, resulting in tumor cell killing. Clinical experience is scarce in MM and restricted to a few patients that have received anti-BCMA (and anti-CD19) CARTs. In contrast, moAbs are about to enter daily practice with potentially huge impact on MM care. The anti-SLAMF7 moAb, elotuzumab, is administered in conjuction with lenalidomide and was shown to significantly prolong event-free survival in relapsed MM in a randomized study. The anti-CD38 moAb, daratumumab, exerts remarkable single agent activity. It may be combined with various other drugs in order to enhance its anti-MM response and is undergoing a comprehensive trial program. Disclosure: Stefan Knop: Advisory Role: Celgene, Janssen, Bristol-Myers Squibb; Financing of Scientific Research: Celgene, Janssen, Bristol-Myers Squibb

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Wissenschaftliches Symposium

Stammzellbiologie als Grundlage für therapeutische Interventionen V809

Progressive epigenetic programming during B cell maturation yields a continuum of disease phenotypes in chronic lymphocytic leukemia Oakes C.1,2, Seifert M.3, Assenov Y.1, Gu L.4, Przekopowitz M.3, Ruppert A.2, Wang Q.5, Serva A.6, Koser S.5, Brocks D.1, Lipka D.1, Bogatyrova O.1, Zapatka M.6, Mertens D.7,8, Lichter P.6, Döhner H.8, Küppers R.3, Zenz T.9, Stilgenbauer S.8, Byrd J.2, Plass C.1 DKFZ Heidelberg, Epigenomik und Krebsrisikofaktoren, Heidelberg, Germany, The Ohio State University, Department of Internal Medicine, Division of Hematology, Columbus, United States, 3Universität Duisburg-Essen, Institut für Zellbiologie, Essen, Germany, 4Broad Institute, Boston, United States, 5 DKFZ Heidelberg, Angewandte Bioinformatik, Heidelberg, Germany, 6DKFZ Heidelberg, Molekulare Genetik, Heidelberg, Germany, 7DKFZ Heidelberg, Mechanismen der Leukämogenese, Heidelberg, Germany, 8Universitätsklinikum Ulm, Innere Medizin III, Ulm, Germany, 9DKFZ und NCT Heidelberg, Molekulare Therapie in der Hämatologie und Onkologie, Heidelberg, Germany 1 2

Charting differences between tumor and normal tissues is a mainstay of cancer research. Yet, as malignant cells typically expand clonally and normal tissues retain complex mixtures of various cell types, reported cancer-specific events are potentially inaccurate. Using whole genome bisulfite sequencing and other methods, we assess the dynamic DNA methylation events that occur as part of a broad epigenetic program established during the maturation of B cells. In comparison to malignant B cells from 268 chronic lymphocytic leukemia (CLL) patients, we reveal that tumors have the potential to derive from a continuum of possible maturation states that are reflected in the maturation stages of normal cells. The degree of maturation achieved in tumors closely associates with the acquisition of a more indolent pattern of gene expression and increasingly favorable clinical outcomes. We further uncover that nearly all methylation differences previously reported between tumor/normal or between defined disease subtypes are naturally present in healthy, non-malignant B cells. Instead, we expose a novel pathogenic role of transcription factor (TF) dysregulation in CLL, where excess activity of NFAT and EGR and loss of EBF and AP-1 TF families imbalances the normal B cell epigenetic program. Disclosure: No conflict of interest disclosed. V810

Genomic profiling of (leukemic) stem cells Bullinger L.1 Klinik für Innere Medizin III, Universitätsklinikum, Ulm, Germany

Introduction: Over the last years, we have learned that with increasing age there is a growing incidence of clonal hematopoiesis, which involves genes known to play a role in hematological malignancies. In addition, it is already known for several years that leukemia is characterized by the sequential acquisition of gene mutations in a single clone of cells and that in acute myeloid leukemia (AML), the acquisition of the first mutation must have occurred in functionally normal hematopoietic stem/progenitor cells (HSPCs). Methods: Within this review, an overview will be provided of the current state of knowledge on genomic profiling of both normal and leukemic HSPCs and findings will be put into perspective with regard to mechanisms of leukemogenesis and future therapeutic relevance. Results: In HSPCs, the majority of somatic mutations occurs in genes such as DNMT3A, TET2, and ASXL1, well-established drivers of leukemogenesis, and the presence of clonal hematopoiesis was shown to be associated with an increased risk of developing a hematologic malignancy. In accordance, respective preleukemic DNMT3A mutant HSPCs were

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shown to have a multilineage repopulation advantage over non-mutated HSCs and their detection in complete remission samples indicated their potential to survive chemotherapy. Furthermore, detailed longitudinal genomic analyses of matched MDS/secondary AML or AML/normal HSPC/relapse AML cases have provided additional insights into the clonal architecture and clonal evolution of leukemia. Conclusion: Future studies will now have to pay further attention to somatic mutations acquired during the process of HSPC aging, and it will be very interesting to see, whether additional preexisting age-related mutations can be identified, which confer a competitive advantage to HSPCs following selection pressure. Finally, in order to answer more open questions, novel genome-wide single cell sequencing based approaches will be of great importance, especially to further unravel the sequence and complex interplay of gene mutations in HSPCs. Disclosure: No conflict of interest disclosed.

Fortbildung

Strahlung und Hämatologie V812

Radiation and Hematology Gale R.P.1 Imperial College of London, Hematology Research Centre, Division of Experimental Medicine, London, United Kingdom 1

Tschernobyl and Fukushima nuclear power facility (NPF) accidents released large amounts of radionuclides into the environment. However, because specifics of the accidents differ substantially, potential impacts on human health are dissimilar. 2 radionuclides are of special concern for human health: 131I and 137Cs. Releases of these radionuclides were 5-10fold greater from Tschernobyl than Fukushima accidents. Howvere, >90% of released radionuclides from Tschernobyl was deposited in populated areas whereas >80% of radionuclides released from Fukushima were deposited in the sea. Tschernobyl accident caused large amounts or 131I to enter the food chain contributing to about 7000 cases of thyroid cancer, predominately in persons < 16 years. There was little food chain exposure to 131I after Fukushima so few if any cases of thyroid cancer are expected. Other heath issues potentially related to radiation exposure are other cancer, cardio-vascular disease, genetic abnormalities and birth defects. There is 1 report of an increased risk of CLL after the Tschernobyl NPF accident amoungst the about 100,00 persons involved in mitigating the accident. These data are not convincing and may reflect surveillance biases. There is no reported increase in the other leukaemias more-convincingly radiation-related such as AML, CML and ALL. Also, there are no convincing data of an increase in any other cancer but it may be too early to exclude this possibility. Using statistical modeling, which may be inappropriate, one might anticipate 11,000-25,000 excess cancers over 70 years in the Northern hemisphere, an incidence undetectable given the background risk of cancer in normals of about 40 percent. There are no convincing data of an increased risk of leukaemia or any other cancer after Fukushima but again this is too early to evaluate critically. On a statistical modeling basis 0-1,500 excess cancers might be anticipated over the next 70 years which would again be undetectable. No persons at Tschernobyl or Fukushima received a radiation dose able to increase risk of cardio-vascular disease and there are no convincing data of genetic abnormalities or birth defects in exposed persons or their progeny from either accident. These relatively mild direct medical outcomes do not consider the considerable physical, psychological, economic and environmental impacts of these accidents which are to be avoided or minimized if possible. Disclosure: No conflict of interest disclosed.

Abstracts

V813

Therapy-related myeloid neoplasms following treatment with radioiodine Schroeder T.1, Kuendgen A.1, Strupp C.1, Haas R.1, Germing U.1 Heinrich-Heine University Düsseldorf, Department of Hematology, Oncology and Clinical Immunology, Düsseldorf, Germany 1

Therapy-related myeloid neoplasms (t-MN) are considered a separate entity within the myeloid stem cell disorders. They have in common the history of an exposition to either mutagenic compounds and/or ionizing radiation. In comparison with de-novo myeloid malignancies the t-MN present more often with chromosomal aberrations and a worse prognosis. The majority of the patients has been exposed to alkylating agents, topoisomerase II inhibitors and antimetabolites with or without radiation, but a subset of the patients was exposed to treatment with radioiodine for a maligne or benign disease only. Numerous case reports were published, summarized by Oluwasnajo et al (Cancer causes control, 2016) and another 39 pts including tMDS, tAML, and tMPN/MDS were published in 2011 by the German MDS study group (Schroeder et al, Haematologica, 2011). 68% of the patients presented with abnormal karyotypes, showing a large variability of aberrations, including mutations of FLT3, NPM1, CEBPA. The median time from iodine exposure to diagnosis of the t-MN was 79 months, which is longer as compared to chemotherapy induced t-MN. The median survival from diagnosis of the t-MN was 29 in tMDS and 12 months in tAML, only, although about 2/3 of the patients were treated with induction chemotherapy or allografting. The prognosis of t-MN primarily influenced by the type of chromosomal findings. Patients with therapy-related promyelocytic leukemia for example have a similar prognosis as compared to the primary counterparts. The large data set of the International MDS study group studying t-MN hopefully will develop prognostic tools that are useful to better stratify patients for different therapeutic interventions. In summary, therapy-related myeloid neoplasms comprise a heterogeneic group of patients with a poor prognosis. Treatment with radioiodine is associated with a certain risk to induce stem cell malignancies and therefore has to be considered carefully. Disclosure: No conflict of interest disclosed. V815

Health effects in German uranium miners: The Wismut cohort study Kreuzer M.1, Sobotzki C.2, Schnelzer M.2 Bundesamt für Strahlenschutz, Fachbereich Strahlenschutz und Gesundheit, Neuherberg, Germany, 2Bundesamt für Strahlenschutz, Neuherberg, Germany 1

Introduction: The aim of the German uranium miner cohort study, which includes 58.972 former workers of the “WISMUT” Company, is to investigate health effects related to occupational exposure to ionizing radiation. A particular focus is given here on radiation-related risk of leukemia. Methods: In the mortality follow-up period from 1946 to 2013, 29.751 miners died, 27.272 are alive and 1.951 lost to follow-up. Overall, 9.066 cancer deaths occurred. Annual exposure to radon, external gamma radiation and long-lived radionuclides was estimated by a comprehensive job-exposure-matrix. In order to evaluate the radiation-related risk of leukemia, additionally absorbed doses to the red bone marrow (RBM) from low-LET (Linear energy transfer) (mainly external gamma radiation) and high-LET (mainly radon gas) radiation in mGy was estimated by dosimetric/biokinetic models. Linear excess relative risks (ERR) per unit of cumulative exposure with 95% confidence limits were estimated by Poisson regression models. Results: There is clear evidence of a linear exposure-response relationship between lung cancer death (n = 3.942) and cumulative radon exposure, which is strongly modified by time since exposure, age at exposure and exposure rate. There is suggestive evidence for an excess of deaths from cancers of the extra-thoracic airways (n = 280) in relation to radon. Overall 203 leukemia deaths occurred, among them 120 deaths from leukemia excluding chronic lymphatic leukemia (CLL), 70 from CLL and 13 from

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unspecified leukemia. The mean absorbed doses to the RBM from lowand high-LET radiation were 47.9 mGy and 8.9 mGy, respectively. There was a positive non-significant dose-response relationship for mortality from non-CLL in relation to low-LET (ERR/Gy = 2.18; 95% CI: -0.41; 6.36) and high-LET radiation (ERR/Gy = 16.69; 95%: -1.10; 46.82). A statistically significant excess was found for the subgroup chronic myeloid leukemia (n = 31) in relation to low-LET radiation (ERR/Gy = 7.20; 95% CI: 0.48; 24.54) and the subgroup myeloid leukemia (n = 99) for high-LET radiation (ERR/Gy = 26.07; 95% CI: 2.58; 69.08). Results indicate no association of death from CLL with neither type of radiation. Conclusion: An increased risk of lung cancer by radon was confirmed in the Wismut cohort. In addition, findings support an increased risk of death from non-CLL in relation to chronic low-LET and high-LET radiation, particularly for myeloid leukemia subtypes and no such relation for CLL. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium

Betreuung und Rehabilitation chronischer Krebspatienten V817

Longterm Survivor and return to work. Do we need new ways? Seifart U.1 Klinik Sonnenblick, Marburg, Germany

Due to improving treatment options patients survive their cancer. Therefore the number of so-called long-term survivors is steadily increasing. Current studies assume that more than 60% of adults survive their cancer. These patients have a variety of problems (chemotherapy-induced neuropathy, fatigue, etc.) that require very special care. In addition to somatic and psychic side effects of the cancer or its treatment, these patients also suffer from the financial and social consequences of the disease. Thus, for long-term survivor in working age, the question of the “return to work” represents a significant problem, since the financial existence of the patient and often the family may substantially dependent. Studies show that cancer patients classify financial worries with regard to the quality of life for more significant than physical or psychological side effects of the cancer or its treatment. Furthermore, there are initial studies which show that the social descent due to the disease, may also forecast relevant for cancer prognosis. In contrast, it was shown in studies and in a Cochrane analysis that professional assistance and support services are suitable for keeping patients in their professional lives.The previously standard practice that patients will be advised at the moment in which they apply for a pension or lose their jobs, possible assistance will take place with a significant time delay.For this reason it seems important to identify “risk-patients” with special professional problems at an early stage of treatment and to offer assistance. Solutions will be discussed during the lecture. Disclosure: No conflict of interest disclosed.

Fortbildung

Betreuung von Heranwachsenden und jungen Erwachsenen (AYA) V820

Long-term toxicities after treatment of childhood cancer Quidde J.1 Universitätsklinikum Hamburg-Eppendorf, Universitäres Cancer Center Hamburg, Hamburg, Germany 1

Multimodality and new treatment approaches offer an increasing number of patients the chance of cure and survival. The last decades have seen tremendous improvements in survival of children, adolescents and young

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adults diagnosed with cancer, with 5-year survival rates approaching 80%. Therefore, the population of cancer survivor is growing. Besides improved efficacy, multimodality treatment increases the risk for physiological, psychological and social long-term sequelae. Complication of cancer treatment may not become apparent until years later. Cancer therapies may have a lasting effect on any organ, e.g. heart, lungs, gastrointestinal tract, kidneys and bladder, skin, eyes, brain, bones and the endocrine and reproductive systems. Adolescents and young adults (AYAs) receiving their cancer treatment during childhood are at particular risk for long-term side effects. According to the Childhood Cancer Survivor study two of three AYAs have treatment related long-term toxicities including learning and memory difficulties, anxiety, depression, hearing loss, cardiac dysfunction, cataracts, obesity, thyroid problems, infertility or second cancer. In addition, an amount of psychosocial issues, like long-term educational, social, behavioural difficulties and facing barriers related to obtaining ongoing medical care, support, and surveillance for late effects are all common. Oeffinger et al compared the health status of 10,397 childhood cancer survivors treated from 1970 to 1987 with 3,034 of their siblings. 62% of the survivors had at least one chronic health condition, and 27% had a serious or life-threatening condition. The major long-term toxicity and cause of mortality after treatment of childhood cancer are cardiovascular diseases like cardiomyopathy, chronic heart failure and valvular problems. Compared to general population AYAs have a 5 to 15-fold increased risk of cardiovascular diseases. Individual risk is determined by treatment related factors (e.g. kind of chemotherapy, way and time of application, number of cycles, cumulative dose of chemotherapy, and combination with radiotherapy) and non-treatment related factors (nicotine abuse, diabetes mellitus, dyslipoproteinemia and hypertension. The talk about “Long-term toxicities after childhood cancer” will be focused on the AYAs at high risk for late effects include patients treated for bone tumors, brain tumors and Hodgkin lymphoma. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium HPV-assoziierte Malignome V823

Basics and diagnostic aspects Dürst M.1 Klinik für Frauenheilkunde, Universitätsklinikum Jena, Jena, Germany

To date over 150 different HPV-genotypes have been identified of which 30 infect the genital tract. On the basis of epidemiological and experimental investigations anogenital HPV types are divided into high risk (hrHPV) and low risk (lrHPV) types. The most prominent representatives are HPV16 and HPV18, which are causative for 70% of all cervical carcinomas. The International Agency for Research on Cancer (IARC) has classified HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59 as carcinogenic for humans (group 1) whereas HPV 68 is probably (group 2A) and HPV26, 53, 66, 67, 70, 73 and 82 are possibly carcinogenic (group 2B). Several other anogenital HPV types such as HPV6 and HPV11 do not possess carcinogenic potential (group 3). Besides cervical carcinomas which are almost exclusively HPV-induced, HPV-infections are causative for up to 50% of further anogenital cancers (vulva, vagina, anus and penis) and for up to 40% of oropharyngeal carcinomas. Independent of the tumor location only a fraction of HPV infections actually result in cancer. This is a clear indication that genetic and epigenetic alterations are also required for carcinogenesis. Thus, a persistent hrHPV infection is a prerequisite but not sufficient cause of cancer. Normal cells underlie numerous, finely tuned concerted growth stimulating and inhibitory signals. The viral oncoproteins E5, E6 and E7 interact with or indirectly influence many cell regulatory proteins and processes. These interactions

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result in genetic instability of the host cell and in phenotypic alterations such as loss of contact inhibition, deregulated proliferation and differentiation, invasive growth, angiogenesis and the ability to metastasize. From a diagnostic point of view, HPV infections can readily be detected by the use of DNA-, RNA- and protein assays. However, when using PCR-based technologies clinically relevant cut-off levels are essential. The detection of only a few HPV molecules in a sample needs to be interpreted critically. This is particularly the case when considering a role of hrHPV in cancers of the oesophagus, prostate, bladder, breast and lung. Disclosure: No conflict of interest disclosed. V825

HPV – Impact and therapeutic implications in head and neck tumors Maschmeyer G.1 Klinikum Ernst von Bergmann, Hämatologie, Onkologie und Palliativmedizin, Potsdam, Germany 1

Among squamous cell carcinomas of the head and neck (HNSCC), human papilloma virus (HV)-induced oropharyngeal cancers represent a distinct entity. At present, around 30 to 40% of oropharyngeal carcinomas are HPV-associated, however, their proportion is increasing in many developed countries due to the declining number of heavy tobacco smokers. HPV-induced HNSCC have a molecular signature different from HPV-negative HNSCC, showing lower mutational load along with specific aberrations, the latter potentially representing the basis of molecular targeted treatment approaches. HPV-induced HNSCC have a better prognosis than HPV-negative HNSCC, as they are more sensitive to chemotherapy and to radiation. First results of studies using a reduced-intensity radiotherapy and cetuximab show a progression-free survival of more than 90% in subgroups of patients with excellent response to chemotherapy. Intermediate- and longterm sequelae of intensive radiotherapy and platinum-based chemotherapy are likely to be significantly attenuated as well. Current (May 2016) evidence-based guidelines do not yet recommend to separate HPV-induced from HPV-negative HNSCC, so that treatment approaches aiming at less intensive treatment in HPV-positive HNSCC are still restricted to controlled clinical studies. Disclosure: No conflict of interest disclosed.

Methods: To analyze the oncogenic potential of Gpr56 and its cooperative affect with Hoxa9, a retroviral overexpression system beside shRNA mediated depletion in combination with a murine bone marrow transplantation model was used. Additionally, to study GPR56 for its potential as a therapeutic target in the human system a xenograft transplantation model was employed. Results: This study demonstrates particularly high expression of GPR56 in patients with NPMc+/FLT3-ITD+ AML and lowest expression in CBF leukemia. Also, high GPR56 expression was associated with inferior prognosis in a large patient cohort treated in two independent multicenter phase III trials. Functional relevance of GPR56 expression was validated in mice. For this, Gpr56 and Hoxa9 were co-overexpressed in bone marrow progenitors and transplanted into mice. After a median time of 148 days post transplantation (range 93 - 264) the mice overexpressing Hoxa9 and Gpr56 developed leukemia. In contrast, the Hoxa9 control did not develop any sign of disease in the observation period of up to 380 days post transplantation. Vice versa, the onset of leukemia was significantly delayed by shRNA-mediated knockdown of Gpr56 in Hoxa9/Meis1 transduced cells, indicating functional relevance of Gpr56 expression in AML. As GPR56 is a surface protein and therefore accessible for blocking antibodies we analyzed the impact of antibody -mediated GPR56 blockage on engraftment of primary human AML cells: treatment with the antibody reduced engraftment by up to 68% in primary AML patients compared to a control isotype antibody. Conclusion: Taken together, GPR56 expression is highly expressed in NPMc+/FLT3-ITD+ AML, is associated with poor prognosis and is functionally relevant. First data demonstrate that antibody-mediated blockage of surface GPR56 interfere with AML growth in vivo. Disclosure: No conflict of interest disclosed. V831

Disturbance of the C/EBPα-miR-182 balance impairs granulocytic differentiation and promotes development of acute myeloid leukemia Wurm A.A.1, Zjablovskaja P.2, Kardošová M.2, Gerloff D.1, BräuerHartmann D.1, Katzerke C.1, Hartmann J.-U.1, Fricke S.3, Hilger N.3, Müller A.-M.3, Bill M.1, Schwind S.1, Tenen D.G.4,5, Niederwieser D.1, Alberich-Jorda M.2, Behre G.1 University of Leipzig, Division of Hematology and Oncology, Leipzig, Germany, Academy of Sciences of the Czech Republic, Institute of Molecular Genetics, Praque, Czech Republic, 3Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany, 4National University of Singapore, Cancer Science Institute, Singapore, Singapore, 5Harvard Medical School, Harvard Stem Cell Institute, Boston, United States 1 2

Freier Vortrag

Akute myeloische Leukämie – experimentell V830

GPR56 contributes to the development of acute myeloid leukemia in mice Kirsten N.1, Daria D.1, Muranyi A.1, Mulaw M.1, Ihme S.1, Kechter A.1, Bullinger L.2, Döhner K.2, Döhner H.2, Feuring-Buske M.2, Buske C.1 University Hospital Ulm, Institute for Experimental Cancer Research, Comprehensive Cancer Center, Ulm, Germany, 2University Hospital Ulm, Department of Internal Medicine III, Ulm, Germany 1

Introduction: G-protein coupled receptor 56 (Gpr56) is an adhesion molecule which collaborates with the extracellular matrix. In the hematopoietic hierarchy, GPR56 expression is highest in lymphoid-primed multipotential progenitors (LMPPs) and decreasing in expression with differentiation. Patients with high GPR56 expression show an inferior overall survival in cytogenetically normal acute myeloid leukemia (CNAML). GPR56 was identified as part of the molecular signature of functionally validated leukemic stem cells isolated from patients with acute myeloid leukemia. The aim of this study was to determine the role of Gpr56 in AML.

Abstracts

Introduction: Silencing of major myeloid transcription factor C/EBPα by gene mutation, promoter hypermethylation or posttranslational modifications is well described and occurs in ~50% of acute myeloid leukemia (AML) cases. Deregulation of C/EBPα by microRNAs, a class of small non-coding RNAs, as a substantial event during AML development or during myeloid differentiation has not been studied yet. Methods: We screened for C/EBPα dependent miRNAs in inducible K562-C/EBPα-ER cells using Illumina’s Next Generation Sequencing. For in vitro functional studies including gain-of-function and loss-offunction experiments, we utilized common acute myeloid leukemia cell lines, human hematopoietic stem cells from umbilical cord blood, murine hematopoietic stem cells from mouse bone marrow and primary human cells from patients with acute myeloid leukemia. For in vivo investigations, we manipulated Lin-Sca-1+c-Kit+ (LSK) murine hematopoietic progenitor cells by lentiviral infection and transplanted them into lethally irradiated littermates. The resulting phenotype was analyzed by flow cytometry and morphological staining of blood and bone marrow. Results: We identified oncogenic miR-182 as strong regulator of C/EBPα during myeloid differentiation and in AML. Moreover, we uncovered a novel regulatory loop between C/EBPα and miR-182. While C/EBPα blocks miR-182 expression by direct promoter binding during myeloid

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differentiation, enforced expression of miR-182 leads to reduced C/EBPα protein levels and impairs granulopoiesis in vitro and in a transplantation based mouse model in vivo. In contrast to this, a knockdown of miR-182 expression enhances C/EBPα protein levels in human AML. Furthermore, we observed highly elevated miR-182 expression levels particularly in AML patients with C-terminal CEBPA mutations and thereby uncovered a mechanism how C/EBPα blocks miR-182 expression. Finally by evaluation of TCGA database, we identified miR-182 expression as a strong adverse prognostic factor in cytogenetically high risk AML patients. Conclusions: Taken together, our data demonstrate the importance of a controlled balance between C/EBPα and miR-182 for the maintenance of healthy granulopoiesis and might uncover a novel mechanism for potential treatment strategies in AML. Disclosure: No conflict of interest disclosed. V832

Different impact of expression levels of IGFBP2 and IGFBP7 on survival and relapse rate in acute promyelocytic leukemia Doll S.1, Nowak D.1, Nowak V.1, Obländer J.1, Xanthopoulos C.1, Büchner T.2, Spiekermann K.3, Hofmann W.-K.1, Lengfelder E.1, Hecht A.1 Universitaetsklinikum Mannheim, Hämatologie und Internistische Onkologie, Mannheim, Germany, 2Universität Münster, Hämatologie/Onkologie, Münster, Germany, 3Ludwig-Maximilians-Universität München, Hämatologie/Onkologie, München, Germany 1

Introduction: High levels of Insulin-like growth factor binding proteins (IGFBP) 2 and 7 are associated with inferior prognosis in different acute leukemias. For acute promyelocytic leukemia (APL), a distinct subtype of acute myeloid leukemia, increased expression of IGFBP2 was reported but no data exist on its impact on prognosis. In addition, no data exist on IGFBP7 in APL. The aim of this study was to elucidate the impact on prognosis of both genes in APL patients. Methods: Expression levels of IGFBP2 and 7 were analysed in bone marrow cells from 22 healthy volunteers and 69 APL patients (time: initial diagnosis; 42 female, 27 male; median age 46 years) that were diagnosed and treated uniformly in the German AML Cooperative Group (AMLCG) studies. Multiplex reverse transcriptase quantitative real-time PCR was performed on a LightCycler® 480 (Roche) PCR system using Glucose-6-phosphat isomerase as a housekeeping gene and K562 cell cDNA as a calibrator. Results were calculated using Pfaffl’s delta-delta CT calculation model. IGFBP2 expression groups were defined as follows: IGFBP2 expression below or equal the 25th percentile (IGFBP2low) versus higher IGFBP2 expression (IGFBP2high). Overall survival (OS), relapse free survival (RFS) and the cumulative incidence of relapse (CIR) were compared using the Kaplan-Meier method and a log-rank test (p < 0.05). Results: Expression levels of IGFBP2 did not differ between APL patients and healthy controls. However, there was a significantly higher relapse rate in APL patients with low IGFBP2 expression (CIR: 25% in the IGFBP2low group vs. 5% in the IGFBP2high group; p = 0.04) and accordingly RFS of patients in the IGFBP2low group was inferior (41% vs. 81% in the IGFBP2high group; p = 0.0002). There was also an impact on OS of therapy responders (61% for IGFBP2low vs. 83% for IGFBP2high; p = 0.02). In contrast, IGFBP7 showed significantly lower expression in APL patients compared to healthy controls (p < 0.0001) but using different cutoffs there was no association with patients’ outcome. Conclusion: We identified low IGFBP2 expression as a novel prognostic marker with strong impact especially on relapse rate and RFS in APL. This is surprising as for leukemias only high IGFBP2 expression has been reported to be a negative factor. However, IGFBP2 has been proposed to act as a tumor suppressor through preventing IGF-receptor driven tumorigenesis. Therefore, further investigations are ongoing to clarify its role in acute leukemia. Disclosure: No conflict of interest disclosed.

V833

FLT3-D835Y-expression in NPM1c bone marrow cells induces an aggressive MPN in mice Rudorf A.1, Mueller T.A.1, Klingeberg C.1, Vassiliou G.S.2, Bradley A.2, Duyster J.1,3, Illert A.L.1,3 Uniklinik Freiburg, Innere Medizin/ Hämatologie, Onkologie und Stammzelltransplantation, Freiburg, Germany, 2Haematological Cancer Genetics, Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom, 3Deutsches Konsortium für Translationale Krebsforschung (DKTK), Heidelberg, Germany 1

Mutations in FLT3 and NPM1 are the most common co-occuring alterations in AML. The Npm1 mutation (NPM1c) leads to a disruption of the nuclear localization signal, dislocating NPM1c to the cytoplasm. In FLT3, two types of mutations are present: Tandem duplications of the juxtamembraneous domain (ITD) and point mutations of the tyrosine kinase domain (TKD). Both NPM1c and FLT3-ITD induce an MPN in murine models, while FLT3-TKD is not sufficient to cause a myeloid disease. Co-expression of NPM1c and FLT3-ITD rapidly induces an AML in C57Bl/6 mice. Here we investigated the impact of co-expression of NPM1c with the TKD mutation FLT3-D835Y in a mouse model. For this purpose, a heterozygous conditional Npm1c knockin mouse model was used. Bone marrow cells from these mice were harvested, retrovirally infected with the MiG-Flt3-D835Y vector and injected into lethally irradiated C57Bl/6 wt mice. Npm1c expression was induced after transplantation and moribund mice were analyzed for their immune phenotype using lineage-specific cell surface markers by flow cytometry. Strikingly, Npm1c Flt3-D835Y mice rapidly developed an MPN with a median latency of 33 days, whereas Flt3-D835Y Npm1 wt mice did not develop a leukemic disease. Detailed analysis of Npm1c Flt3-D835Y mice revealed a high leukemic burden in the peripheral blood (PB) and bone marrow (BM) as well as a pronounced splenomegaly. Flow cytometry analysis indicated a high frequency of FLT3-D835Y+/CD11b+ and Gr1+ cells in the PB, spleen and BM, whereas Flt3-D835Y control mice showed a normal immune phenotype. Further analyses of the Npm1c Flt3-D835Y induced MPN revealed the retransplantability of the disease in < 80% of the recipient mice transplanted with 1×105 and 1×106 Npm1c Flt3-D835Y donor spleen cells. Interestingly, immunoblot analysis of spleen extracts from primary transplanted, moribund mice indicated that FLT3-D835Y in combination with NPM1c induces STAT5 but not STAT3 signaling. These results suggest that the MPN induction in this model might be due to STAT5 activation. Our data thus show that co-expression of Npm1c and Flt3-D835Y results in a rapidly fatal MPN whereas Flt3-D835Y expression alone is not sufficient to cause a myeloproliferative disease in C57Bl/6 mice. Moreover, the myeloid phenotype is retransplantable with 1×106 donor cells. Interestingly, the phenotype switch leading to a myeloproliferative disease might be due to aberrant STAT5 activation by FLT3-D835Y in the NPM1c background. Disclosure: No conflict of interest disclosed. V834

PML/RARα-regulated miR-181a/b-cluster targets the tumor suppressor RASSF1A in acute promyelocytic leukemia Bräuer-Hartmann D.1, Hartmann J.-U.1, Wurm A.A.1, Gerloff D.1, Katzerke C.1, Verga Falzacappa M.V.2, Pelicci P.G.2, Müller-Tidow C.3, Tenen D.G.4,5, Niederwieser D.1, Behre G.1 University Leipzig, Division Onkology/Hematology, Leipzig, Germany, European Institute of Oncology, Department of Experimental Oncology, Milan, Italy, 3University Hospital Halle, Department of Internal Medicine IV, Hematology/Oncology, Halle, Germany, 4Harvard Medical School, Harvard Stem Cell Institute, Boston, United States, 5National University of Singapore, Cancer Science Institute, Singapore, Singapore 1 2

Introduction: In acute promyelocytic leukemia (APL), carrying the PML/ RARα oncogene, all-trans retinoic acid (ATRA) treatment induces gran-

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ulocytic maturation and complete remission of leukemia. Several studies identified microRNAs as critical players in the formation of the leukemic phenotype. Methods: In this study, we used AML cell lines, AML patient samples and a PML/RARα-knock in-mouse model for analyzing microRNA and protein levels. In vitro functional studies were performed by lentiviral knock down, gene overexpression and microRNA mimic transfection. Analysis was done by replating assay, apoptosis assay, proliferation assay, cell cycle analysis and FACS measurement of granulocytic surface markers. Luciferase assay was used to prove direct microRNA binding to the 3´UTR of the target mRNA. Results: Our data show the downregulation of the miR-181a/b gene cluster in APL blasts and NB4 leukemia cells upon ATRA treatment as key event in the drug response, which seems highly specific for APL. We found that miR-181a/b expression was activated by the PML/RARα oncogene in cells and transgenic knock-in mice. This observation was confirmed and extended by evidence of enhanced expression of miR-181a/b in APL patient specimens. Functional studies showed that miR-ZIP-mediated attenuation of miR-181a/b expression in NB4 cells caused reduced colony forming capacity, proliferation and survival. Mechanistic investigations revealed that miR-181a/b targets the novel ATRA-regulated tumor suppressor gene RASSF1A by direct binding to its 3´UTR. Enforced expression of miR-181a/b or RNAi-mediated attenuation of RASSF1A inhibited ATRA-induced granulocytic differentiation via regulation of the cell cycle regulator cyclin D1. Conversely, RASSF1A overexpression leads to enhanced apoptosis in NB4 cells. Lastly, RASSF1A levels were reduced in PML/RARα knock-in mice and APL patient samples. Conclusion: Taken together, our results could define miR-181a and miR181b as oncomiRs in PML/RARα-associated APL. Furthermore, the miR181a/b target RASSF1A could be identified as a pivotal element in the ATRA-induced granulocytic differentiation program in APL. Disclosure: No conflict of interest disclosed. V835

The homeobox gene VENTX contributes to the development of acute myeloid leukemia with erythroid features in mice Gentner E.1,2, Vegi N.1, Mulaw M.A.1, Quintanilla-Fend L.3, Döhner H.4, Döhner K.4, Buske C.1, Rawat V.P.S.1, Feuring-Buske M.4 University Hospital of Ulm, Institute of Experimental Cancer Research, Ulm, Germany, 2Ulm University, International Graduate School in Molecular Medicine Ulm, Ulm, Germany, 3Institute of Pathology, Eberhard Karls University, Tuebingen, Germany, 4University Hospital of Ulm, Department of Internal Medicine III, Ulm, Germany 1

Introduction: Acute myeloid leukemia (AML) with erythroid features is rare and molecular events inducing this AML phenotype are poorly characterized. Our group has previously shown that the Vent-like homeobox gene VENTX shows an aberrantly high expression in human AML characterized by the AML1-ETO (AE) fusion (Rawat et al., PNAS 2010). We now demonstrate that overexpression of VENTX expands primitive erythroid cells, has leukemogenic potential and the ability to induce AML with erythroid features in transplanted mice. Methods: For this purpose mice were transplanted with bone marrow progenitor cells retrovirally transduced to express VENTX and/or AML1ETO. Results: When mice were transplanted with bone marrow cells overexpressing VENTX, massive expansion of the primitive erythroid compartment was observed (8 of 8 transplanted mice) with a conversion into an acute erythroleukemia in 4 of 8 mice. All recipient mice transplanted with AE+VENTX double transduced cells succumbed to rapidly re-transplantable AML after a median latency of 354 (80 - 403) days post transplantation (n = 20). In contrast, none of the AE mice developed any disease (n = 10). In line with the observations in VENTX diseased mice AE+VENTX AML expressed the erythroid markers CD71 and Ter119 and showed an erythroblastic phenotype. Incubation of this cell population with EPO or hemin led to a differentiation of erythroblasts, as shown

Abstracts

by a loss of Ter119 and an acquisition of CD71 expression measured by flow cytometry. RNA-Seq of human cord blood (CB) cells retrovirally engineered to express VENTX (n = 3) showed an enrichment for pathways such as “Jak-STAT signaling pathway”, “Hematopoietic cell lineage”, “Cytokine-cytokine receptor interaction” and “Hemoglobin’s chaperone” (KEGG analysis). Genes necessary for terminal erythroid differentiation such as the EPO-receptor, GATA1, GATA2, KLF4 were significantly downregulated upon constitutive VENTX expression. VENTX was highly expressed in patients with primary human erythroleukemias (n = 10) and shRNA mediated knockdown of VENTX impaired in vitro and in vivo growth in NSG mice of the erythroleukemic HEL cell line. Conclusion: In summary, these data indicate that aberrant expression of VENTX induces expansion of primitive erythroid cells and is able to induce AML with erythroid features in mice. Ongoing analyses will dissect the role of this non-clustered homeobox gene in human acute erythroleukemia. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Tumor-/Zellbiologie V838

Mast cell-derived granzyme b mediates resistance against anti-angiogenic therapy Wroblewski M.1,2, Bauer R.1,2, Cubas Córdova M.1,2, Udonta F.1,2, Ben Batalla I.1,2, Gensch V.1,2, Sawall S.1,2, Waizenegger J.1,2, Pardo Jimeno J.3, Pantel K.2, Bokemeyer C.1, Loges S.1,2 Department of Hematology and Oncology with Sections BMT and Pneumology, Hubertus Wald Tumorzentrum, University Comprehensive Cancer Center Hamburg, University Medical Center Hamburg-Eppendorf, Hamburg, Germany, 2Department of Tumor Biology, Center of Experimental Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany, 3Instituto de Nanociencia de Aragón, Zaragoza, Spain 1

Introduction: Targeted therapies have revolutionized the treatment of cancer. However, efficacy of anti-angiogenic therapies is limited due to significant resistance. Based on the correlation of mast cell (MC) density with tumor growth and angiogenesis we put forward the hypothesis that MC might be implicated in resistance against anti-angiogenic therapy. Methods: C57BL/6J, NSG or MC-deficient KitW-sh (Wsh) mice were subcutaneously injected with 5×105 (Panc02 and EL4) or 1×106 (TD2) cells ± bone marrow derived MC. Tumors were treated with 20 mg/kg anti-VEGFR2 antibody (DC101) or 25 mg/kg cromoglicic acid. BrdU was injected 12 h before sacrifice. Stainings were performed on FFPE sections. Results: We showed that MC alter the proliferative and organizational state of endothelial cells (EC). MC dose-dependently induced the proliferation, migration and tube formation of EC in vitro. In different tumor models, MC-deficiency sensitized tumors for anti-angiogenic therapy, resulting in reduced microvessel density and tumor cell proliferation. In WT mice, anti-angiogenic therapy only transiently reduced the proliferation of tumor vessels, a process that got reverted after long-term treatment as a result of acquired therapy resistance. Intriguingly, this pro-angiogenic rescue did not occur in MC-deficient mice. The observed “angiosensitization” phenotype was causally related to the absence of mast cells, as adoptive transfer of MC into MC-deficient mice restored resistance. Microarray analysis of tumor-resident MC revealed increased expression of matrix-degrading granzyme b in response to therapy. We showed that MC-specific knock down of granzyme b lowered the levels of alternative, pro-angiogenic mediators beside the VEGF-VEGFR2-axis in the tumor microenvironment, which rendered tumors more susceptible for anti-VEGFR2 therapy. By blocking MC degranulation with the approved mast cell stabilizer cromoglicic acid, we were able to counteract MC-mediated resistance, which

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sensitized tumors for anti-angiogenic therapy and resulted in reduced vessel proliferation, microvessel density and tumor growth. Conclusion: Mast cell-derived granzyme b liberates matrix-bound pro-angiogenic factors besides the targeted VEGF-VEGFR2 axis, which confers resistance to anti-angiogenic therapy. Blockade of mast cell function might therefore represent a novel approach to improve the efficacy of anti-angiogenic drugs in the clinic. Disclosure: Mark Wroblewski: Employment or Leadership Position: Anstellungsverhältnis UKE; Advisory Role: Eli Lilly Sonja Loges: Employment or Leadership Position: Führungsposition UKE; Advisory Role: Eli Lilly; Financing of Scientific Research: Eli Lilly; Expert Testimony: Eli Lilly V839

The niche protects hematopoietic stem cells through cytokines which regulate DNA repair and cytoskeleton dynamics Oostendorp R.1, Schreck C.1, Ziegenhain C.2, Sippenauer T.1, Ruf F.1, Vilne B.1, Peschel C.1, Enard W.2, Götze K.1, Istvanffy R.1 Klinikum rechts der Isar der Technischen Universität München, III. Medizinische Klinik und Poliklinik, München, Germany, 2Ludwig-Maximilians Universität München, Anthropology and Human Genetics, Department Biology II, Martinsried, Germany 1

Introduction: The niche is required for the lifetime maintenance of hematopoietic stem cells (HSCs). The main function of HSCs appears to restore the hematopoietic hierarchy after hematopoietic stress, such as toxic insult or transplantation. In addition, alterations in the niche have been shown to collaborate in leukemogenesis and are involved in the diminished HSC function in aging. Methods: In studies of the niche gene products involved in HSC regulation, we determined the effects of deletion of Ctgf, Sfrp1, or Sfrp2 in the microenvironment on hematopoiesis in vitro and in vivo under steady state or stressed conditions. Results: The deletion of Ctgf, Sfrp1, or Sfrp2 in stromal cell line UG261B6, leads to inability to maintain repopulating HSCs in vitro. In respective mutant mice, we show that deletion of these genes shows no major alterations in steady-state hematopoeisis. Nonetheless, transplantation of wild-type HSCs in Sfrp1-/- or Sfrp2-/- mice shows diminished repopulation of donor HSCs. In gene expression studies designed to determine the interplay between HSCs and the niche, we showed upregulation of stromal Ctgf induced by co-cultured HSCs. Interestingly, clonal cell division of HSCs in media from stromal cells deficient in CTGF was defective, which could be rescued by addition of rCTGF. This indicates a direct effet of CTGF on HSCs. Similar effects were found in clonal cultures with conditioned media from stromal cells lacking SFRP1. To find out underlying mechanisms of HSC growth arrest, we found that HSCs grown in CTGF- or SFRP1-deficient media show an accumulation of gH2AX+ heterochromatin foci and acetylated H4K16. In Ctgf-deficient stromal cell media, this is also accompanied by activation of SMAD2/3 and PTEN in HSCs. These markers are frequently found in cellular senescence. In vivo, HSCs from a Sfrp2 deficient environment show similar changes in senescence marker expression after transplantation, 5-fluorouracil treatment or upon aging. Conclusion: Taken together, our studies show that the secreted niche factors CTGF, SFRP1, and SFRP2 are required to maintain DNA stability. Lack of these molecules results in increased DNA damage, which is associated with delayed cell division and HSCs exhaustion. Our studies offer new targets in CTGF and SFRP1/SFRP2 signaling to prevent cellular senescence in vitro (expansion), or, alternatively, reactivate senescence in malignancies in which alterations allowing senescence evasion have occurred. Disclosure: No conflict of interest disclosed.

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V840

Targeted multiplex re-sequencing identifies loss-of-function uORF mutations in human cancer Wethmar K.1,2, Schulz J.2, Mah N.3, Kischka T.4, Ratei R.5, Schlag P.M.6, Castaños-Vélez E.6, Fichtner I.7, Tunn P.-U.8, Denkert C.9, Klaas O.1, Berdel W.E.1, Makałowski W.4, Andrade-Navarro M.A.10, Leutz A.2,11 University Hospital Muenster, Department of Medicine A, Hematology, Oncology and Pneumology, Muenster, Germany, 2Max-Delbrueck-Center for Molecular Medicine, Department of Cell Differentiation and Tumorigenesis, Berlin, Germany, 3Charité University Medicine Berlin, Campus Virchow-Klinikum, Berlin-Brandenburg Center for Regenerative Therapies, Berlin, Germany, 4 University of Muenster, Faculty of Medicine, Institute of Bioinformatics, Muenster, Germany, 5Carl-Thiem-Klinikum, 2. Medizinische Klinik, Cottbus, Germany, 6Charité Comprehensive Cancer Center, Berlin, Germany, 7MaxDelbrueck-Center for Molecular Medicine, Berlin, Germany, 8Helios Kliniken Berlin, Berlin, Germany, 9Charité University Medicine Berlin, Institute of Pathology, Berlin, Germany, 10Johannes-Gutenberg University of Mainz, Institute of Molecular Biology, Mainz, Germany, 11Humboldt-University, Department of Biology, Berlin, Germany 1

Translational control by upstream open reading frames (uORFs) is a widespread mechanism of regulated gene expression with more than 60% of human transcripts containing one or more AUG-initiated uORF(s). Upstream ORFs commonly repress translation from the associated downstream main protein coding sequence and mutational changes in uORFs have been linked to various diseases, including cancer. Nevertheless, a systematic experimental search for cancer-associated genetic alterations in uORFs has not been performed. Here, we applied a PCR-based, multiplex identifier-tagged deep sequencing approach to screen 404 uORF translation initiation sites (uAUGs) of 83 human tyrosine kinases and 49 other proto-oncogenes in 308 hematologic and solid tissue malignancies. We identified several novel mutations that disrupted uORF initiation codons or altered uORF-related Kozak consensus sequences. Upstream AUG-deleting mutations were detected in the ephrin receptor B1 (EPHB1) in breast and colon cancer and in the mitogen-activated protein kinase kinase 6 (MAP2K6) in a colon adenocarcinoma. Mutant transcripts showed enhanced translation of downstream protein coding sequences by approximately one third for EPHB1 and three-fold for MAP2K6, respectively. Presence of the MAP2K6 transcript was validated in the primary tumor sample and the translational effect of the MAP2K6 loss-of-function uORF mutation was confirmed in colon cancer cells. The data suggest that mutations causing a loss of uORF initiation codons in onco-developmental genes may contribute to malignant transformation and cancer development. In a current genome-wide approach we explore cancer-derived whole genome sequencing data to define the genetic variability for all of the more than 25.000 annotated uAUGs in the human genome. Ultimately, these efforts may uncover recurrent uORF-associated oncogenic driver mutations and may identify novel molecular targets for cancer therapy. Disclosure: No conflict of interest disclosed. V841

Exploring p53-governed pro-senescent mTOR signaling for lymphoma therapy in vitro and in vivo Gerhardt A.1, Dörr J.R.1, Milanovic M.1, Leser U.2, Zimmermann K.2, Dörken B.1,3, Lee S.1,3, Schmitt C.A.1,3 Charité – Universitätsmedizin Berlin (CVK), Berlin, Germany, 2Institute for Computer Science, Humboldt-Universität zu Berlin, Berlin, Germany, 3MaxDelbrück-Center for Molecular Medicine, Berlin, Germany 1

Introduction: Cellular senescence is a permanent G1 cell-cycle arrest which occurs in ageing cells or prematurely in response to acute stresses, e.g. chemotherapy. Therapy-induced senescence (TIS) depends on the tumor suppressor p53, whose target genes Sestrin2 and phlda3 (Pleckstrin homology-like domain family A member 3) modulate mTOR (mammalian target of Rapamycin) activity via activation of AMPKa (AMP-activated

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kinase) and via inhibition of AKT. Here we use the Eµ-myc mouse lymphoma model to investigate whether Sestrin2- and Phlda3-mediated inhibition of mTOR represents important components of the p53-governed senescence program. Methods: p53-proficient (control) and p53-deficient lymphoma cells (LC) and LCs with regulatable p53 activity (p53ERTam) were treated with adriamycin or tamoxifen to induce senescence in vitro. p53 target genes were analyzed by RQ-PCR. Signaling pathways were assessed by Western Blotting. Pharmacological agents, e.g. the AMPK activator Metformin, the AKT inhibitor Triciribine, the mTOR inhibitor Rapamycin, and stable knockdown of the Tuberosis Sclerosis 2 (TSC2) gene product by shRNA were used to dissect the role of the mTOR pathway in senescence. Results: Besides senescence-associated secretory factors, cell-cycle and apoptosis regulators, modulators of the mTOR signaling axis, namely Sestrin2 and Phlda3, were induced by p53 in TIS. They operate in parallel cascades converging at the level of the TSC2 gene product, a negative regulator of mTOR signaling. Consistent with their action downstream of p53, we found that only combined pharmacological activation of AMPKa and inhibition of AKT induced a senescent phenotype in LCs, irrespective of their p53 status. Treatment with Rapamycin induced senescence in both p53-proficient and -deficient settings, suggesting that the above described pathways orchestrate senescence downstream, and, hence, even in the absence of p53. Strikingly, Rapamycin in vivo-administration significantly extended the overall survival of both control and p53null lymphoma-bearing mice with senescence detectable in lymph nodes of Rapamycin treated mice, but not their control counterparts. Conclusion: Senescence is characterized by the activation of a p53-dependent target gene network. Two key components of this network – Sestrin2 and Phlda3 - modulate mTOR activity via TSC2 and thereby operate as critical mediators to senescence distal of p53, rendering them attractive targets also in p53-compromised lymphomas. Disclosure: No conflict of interest disclosed. V842

Deciphering interaction networks of AKT and SOX2 in oncogenesis Schaefer T.1, Candido S.1, Wang H.1, Bock T.2, Schmidt A.2, Lengerke C.1,3 University Hospital Basel, Department of Biomedicine, Basel, Switzerland, University of Basel, Biozentrum, Basel, Switzerland, 3University Hospital Basel, Division of Hematology, Basel, Switzerland 1 2

Introduction: The pluripotency-associated transcription factor SOX2 (sex determining region Y – box 2) is a key regulator of embryonic stem cell fate and fosters reprogramming of terminally differentiated somatic cells into a pluripotent condition. More recently, SOX2 expression was documented also in several cancers and linked to the cancer stem cell (CSC) compartment. We and others have shown that SOX2 expression heavily relies on canonical PI3K/AKT signaling involving direct physical contact and phosphorylation by AKT. Noteworthy, co-factors regulating this interaction have not yet been analyzed. Methods: Various breast (BC: MCF7, T47D, BT474), ovarian (OC: OVCAR3, OVCAR5) and head and neck squamous cell carcinoma (HNSCC: FadU, HSC3, HN, SCC25), as well as glioblastoma cell lines (GB: U373, LN319) were treated with different PI3K/AKT pathway inhibitors (anti-PI3K: Wortmannin, GDCO941, BYL719, BKM120, and anti-AKT: MK-2206, AKTi-1/2) and the effects on expression, localization and turnover of SOX2 protein analyzed by qRT-PCR, IHC, and immunoblot. SOX2 pull-down experiments were performed and co-enriched factors identified by quantitative mass spectrometry, followed by computational network analysis. Results: We found that in BC and HNSCC but not in OC cells, PI3K/ AKT pathway inhibition blocks SOX2 nuclear entry and fosters proteasomal degradation of cytoplasmic SOX2. Glioblastoma samples gave mixed results with an overt dependence of SOX2 stability on AKT activity in LN319, but not in U373 cells. Mirroring these phenotypic differences, IP-analyses confirmed a direct physical association of SOX2 protein (bait)

Abstracts

with AKT and a number of further canonical interaction partners. More interestingly though, a large number of cell-type specific co-factors were pulled down, whose functional relevance is currently tested using small cpd inhibitors and knock-down technology. Conclusion: Our synoptic investigation of cell lines derived from various human tumors identified tissue-immanent differences in the regulation of SOX2 expression, subcellular localization and turnover in dependence of PI3K/AKT signaling. We hypothesize that these phenotypic differences are due to selective expression of a decisive set of tissue-specific co-factors that physically associate with SOX2 and modulate its interaction with AKT. These investigations may help identify tumor entities in which inhibition of AKT signaling can effectively suppress SOX2 and thereby the CSC compartment. Disclosure: No conflict of interest disclosed. V843

Mechanisms of primary and acquired resistance to 3rd generation EGFR TKI treatment in EGFR mutated lung adenocarcinoma Michels S.1, Fischer R.1, Heydt C.2, Ihle M.2, Scheffler M.2, Scheel A.2, Ortiz-Cuaran S.3, Nogova L.1, Brandes V.1, Sos M.2, Büttner R.2, MerkelbachBruse S.2, Wolf J.1, Lung Cancer Group Cologne and Network Genomic Medicine Uniklinik Köln, Klinik I für Innere Medizin, Lung Cancer Group Cologne, Köln, Germany, 2Uniklinik Köln, Institut für Pathologie, Lung Cancer Group Cologne, Köln, Germany, 3Uniklinik Köln, Institut für translationale Genomik, Köln, Germany 1

Introduction: Activating mutations (mut) in EGFR are found in 10-15% of lung cancer patients and treatment with 1st/2nd generation (gen) EGFR TKIs is highly effective in this subgroup. However, resistance to treatment inevitably develops due to selection of drug-desensitized tumour cells. In 60% of cases acquired resistance is mediated through the gate-keeper mutation EGFR p.T790M. Third gen TKIs such as osimertinib, rociletinib and EGF816 have been developed to overcome resistance due to T790M. However, cases of primary resistance are frequent and acquired resistance limits PFS. Methods: At the time-point of data cut-off for this abstract efficacy data was available for 24 patients with EGFR mutant NSCLCs in the acquired resistance setting treated with osimertinib (NCT01802632), rociletinib (NCT02147990) and EGF816 (NCT02108964) within clinical trials. Pre-treatment tumor tissue was analysed for multiple resistance-related genes by MPS and FISH. At time of progression rebiopsies were obtained of 10 patients and analysed for potential mechanisms of resistance to 3rd gen EGFR TKI treatment. Results: The confirmed ORR in all patients was 50% (N = 12; 95% CI 30.869.2). Four patients suffered from primary progression (17%; CI 3.9-34.1). Of those, three had a high level (hl) MET amplification (a), PFS of this patient subgroup was 1.0 months (95% CI 0.5-1.5). PFS of patients with low level MET a was 4.2 months (95% CI 3.8-10.7) as compared to 8.2 (95% CI 2.4-13.9) months in patients with no/undetermined MET a. No difference was seen in the PFS regarding the number of prior EGFR TKIs. Of the 10 patients rebiopsied upon progression, we have found changes of the molecular pattern as compared to baseline in 7. Isolated MET a were observed in four patients, one patient exhibited a KRAS mut and loss of T790M. Polygenetic mechanisms of resistance were found in two patients: EGFR p.C797S plus intermediate level MET a, hl MET plus HER2 a. Conclusions: Molecular-pathological analyses of rebiopsies have revealed a complex pattern of acquired resistance to all classes of EGFR TKIs, involving aberrations in MET, HER2 and MEK/RAS. Our data suggest that NSCLCs harbouring multiple mechanisms of resistance may not respond to treatment with 3rd gen EGFR TKI alone. Most notably, hl MET a is a major factor driving primary and acquired resistance. These findings triggered approaches to treat patients with combinations of TKIs according to the distinct pattern of mechanisms of resistance.

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Disclosure: Sebastian Michels: Advisory Role: Novartis; Financing of Scientific Research: Novartis, AstraZeneca Jürgen Wolf: Advisory Role: Novartis, AstraZeneca, Clovis; Financing of Scientific Research: Novartis, AstraZeneca, Clovis

Posterdiskussion

Debatten

MRD-Negativität: Ein klinisch relevantes Therapieziel bei der CLL?

Targeting Kv1.3 potassium channels as a novel strategy to enhance the efficacy of cytarabine treatment in acute leukemia

V846

Lowinus T.1, Heidel F.2,3,4, Bose T.5,6, Schnöder T.3,4, Schmitz I.1,7, Seifert U.1,8, Fischer T.2, Schraven B.1,9, Bommhardt U.1

Akute lymphatische Leukämie, chronische lymphatische Leukämie P847

Contra: MRD-negativity as goal of treatment in CLL Knauf W.

Centrum für Hämatologie und Onkologie Bethanien, Frankfurt, Germany

Much progress has been achieved in the treatment of chronic lymphocytic leukaemia. Along with advantages in supportive care, e.g. the availability of blood products and effective anti-infectious agents, it is the development of new drugs that provides the patients with a better control over the disease. Coming from mono-therapy with Chlorambucil (CBL), nowadays, combined regimens based on Fludarabine (F) or Bendamustine (B) with antiCD20 monoclonal antibodies result in high overall remission rates including complete remissions (CR) even at the molecular level. Currently, assessment of minimal residual disease (MRD) is part of the protocol in many trials, however, methods are not standardized and therefore, CR may act as surrogate parameter. Prolonged progression free survival (PFS) times were shown to be linked with CR rates and gave the reason to approve a number of drugs and to define so-called standards of care. But, what does this mean with respect to overall survival (OS) which is the ultimate goal of any therapy? In a British land-mark study, OS was not shorter with CBL as compared with F or FC even though CR rates were remarkably greater with the latter treatments. In contrast, depth of remission was associated with prolonged OS in the CLL 8 trail that compared FCR with FC. While FCR showed higher CR rates and had a probability to achieve MRD negativity higher than with BR, OS was not different between the two regimens. Surprisingly, the inhibitors of signal transduction pathways like ibrutinib or idelalisib were shown to prolong significantly PFS in high risk patients without inducing relevant CR rates. Do we need, in consequence, achieve a CR (more or less equivalent to MRD negativity) at all ? The average CLL patient is aged 70-72 years at first diagnosis. Approximately 30% of the pts will never be in need for specific treatment while co-morbidities are likely to define the life span for the majority. Meanwhile, second line treatments with high efficacy are available. Rendering CLL in a manageable condition with continuous treatment or with repetitive various therapies is a reasonable goal to achieve a life expectancy which is defined by events other than CLL, but does not depend on MRD negativity. Disclosure: Wolfgang Knauf: Advisory Role: Gilead, Janssen, Mundipharma, Roche; Financing of Scientific Research: Gilead, Janssen, Mundipharma, Roche

Institute of Molecular and Clinical Immunology, University Magdeburg, Magdeburg, Germany, 2Department of Hematology and Oncology, University Magdeburg, Magdeburg, Germany, 3Internal Medicine II, Hematology and Oncology, University Jena, Jena, Germany, 4Leibniz-Institute on Aging, Fritz-Lipmann-Institute, Jena, Germany, 5Leibniz-Institute for Neurobiology, Magdeburg, Germany, 6Lee Kong Chian School of Medicine, Singapore, Singapore, 7Systems-oriented Immunology and Inflammation Research Group, HZI, Braunschweig, Germany, 8Friedrich Loeffler Institute of Medical Microbiology, University Medicine Greifswald, Greifswald, Germany, 9 Department of Immune Control, HZI, Braunschweig, Germany 1

Treatment of acute leukemia is still challenging and long-lasting remissions are difficult to induce, especially in elderly or frail patients. Key signaling pathways, such as PI3-K-AKT-mTOR and ERK1/2 cascades, are frequently overactive in acute leukemia, and their inhibition may create synergistic effects with standard chemotherapy. However, compensatory signaling pathways are often induced leading to drug resistance and relapse. Here, we provide first evidence that genetic inactivation or pharmacologic blockade of Kv1.3 potassium channels synergistically enhances cytarabine-induced cell cycle arrest and cell death in lymphoid and myeloid leukemia. Inhibition of Kv1.3 channels by memantine, a clinically approved antagonist of neuronal N-methyl-D-aspartate receptors, reinforces cytarabine-induced down-regulation of AKT-mTOR-S6 and ERK1/2 signaling, fosters silencing of nuclear c-MYC and promotes the release of cytochrome C and activation of Caspase-9. Hence, inactivation of Kv1.3 channels in combination with cytarabine induces simultaneous blocking of central signaling pathways and oncogenic effector molecules, thereby providing an attractive strategy for combination therapy in acute leukemia. Disclosure: No conflict of interest disclosed. P848

Comparative cellular and molecular analysis of single and combined drug application of cytarabine, dexamethasone and idelalisib in B-lymphoblastic leukemia cells Sklarz L.1, Ernst M.2, Roolf C.1, Sekora A.1, Knübel G.1, Struckmann S.2, Du Y.2, Beck J.3, Schütz E.3, Fuellen G.2, Murua Escobar H.1, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany, 2Institute for Biostatistics and Informatics in Medicine and Ageing Research, Rostock, Germany, 3Chronix Biomedical GmbH, Göttingen, Germany 1

Introduction: Treatment of patients with acute lymphoblastic leukemia (ALL) comprises of several cytostatic agents in different application schemes in sequence. The respective combination partners differ between regimens. New drugs as tyrosine kinase inhibitors are introduced thus attention might be necessary to identify best partners and application time points. Combined ALL culture and gene expression based experimental design was used to identify effects of single and combined applications. Along with known drugs such as cytarabine (AraC) and dexamethasone (Dexa) novel drugs as idelalisib (Idel) were tested. Methods: Pre B-ALL cell lines (RS4;11, SEM) were incubated with single AraC, Dexa, Idel or a combination of two (72 h). Drug concentrations were selected allowing a threshold of above 30% living cells after 72 h or a maximum of 10 µM. Effects on proliferation (trypan blue staining) and metabolism (WST-1), apoptosis/necrosis (AnnexinV/PI staining) were

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analyzed. Whole transcriptome analyses were performed with a least 30×106 reads per library. Bioinformatical analyses were carried out via an in-house pipeline (including DeSeq2, WikiPathways, igraph). Experiments were performed at least in biological replica. Results: Single and combined drug application usually resulted in significantly decreased proliferation and metabolic activity. Apoptosis/necrosis rates increased compared to controls. In RS4;11 cells AraC+Idel and Dexa+Idel, and in SEM cells AraC+Dexa showed stronger antiproliferative effects compared to mono application. AraC, Dexa or Idel exposed cells showed characteristic gene expression patterns. Combinations resulted in expression values similar to the higher expression level of the respective mono application treatment groups. Incubation of SEM with AraC+Dexa and RS4;11 with AraC or Dexa with Idel resulted in extraordinary upregulation of selected genes compared to single application values. Some combinations resulted in upregulation of senescence-autophagy pathways as well as downregulation of pathways involved in glycolysis, gluconeogenesis, cell cycle and DNA replication. Conclusion: AraC, Dexa and Idel affect the expression of different genes in pre-B-ALL. In SEM cells, the combination of AraC+Dexa leads to synergy-like effects in gene expression and shows an enhanced antiproliferative effect compared to single drug application in vitro. Similar effects were detected in RS4;11 cells incubated with AraC or Dexa with Idel. Disclosure: No conflict of interest disclosed. P849

Synergistic inhibitory effects of demethylating and cytostatic agents on T-lymphoblastic leukemia cell proliferation Khodamoradi Y.1, Roolf C.1, Richter A.1, Sklarz L.1, Sekora A.1, Knübel G.1, Murua Escobar H.1, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany 1

Introduction: Aberrant methylation of tumorsuppressor gene promotors (e.g. DBC1, ZNF462 and CPNE7) are frequently observed in acute T-lymphoblastic leukemia (T-ALL) cells. Methylation status has been shown to be an independent prognostic factor regarding disease-free survival and plays an important role in drug-resistance. Cytosine analoga Azacitidine (AZA) and Decitabine (DAC) are methyltransferase inhibitors with clinically activity in myelodysplastic syndrome and acute myeloid leukemias. However, it is unknown if aforementioned agents have biological effects on T-ALL cells. Methods: T-ALL cell lines CEM and Jurkat were incubated for 72 h with increasing concentrations of AZA and DAC ranging between 100 - 1000 nM. Cell proliferation and metabolism were examined by trypan blue staining and WST-1 assay, respectively. Methylation status of bisulfite converted DNA samples were examined using methylation specific qPCR. Further, combined applications of DAC (100 nM or 500 nM) with IC80 values of cytarabine, dexamethasone and doxorubicin were analysed. Three application sequences were used: respective cytostatic 24 h before (A), simultaneously (B) or 24 h after DAC application (C). Results: Mono application of DAC starting at concentrations of 500 nM reduced metabolic activity and proliferation significantly in both cell lines (CEM + DAC: 73.0 ± 5.8%, Jurkat + DAC: 74.2 ± 4.8%). Furthermore, DAC reduced the methylation status on LINE 1 element in Jurkat cells: 55.0 ± 4.7% after 48 h and 56.5 ± 2.3% after 72 h compared to DMSO control group (100%). In both cell lines DAC showed synergistic effects with dexamethasone and doxorubicin. In CEM cells, proliferation and metabolic activity were reduced significantly when DAC was combined with dexamethasone: 26.7 ± 1.0% compared to DAC: 65.6 ± 0.8% and dexamethasone: 76.6 ± 4.6%, after 72 h simultaneously. Conclusion: The methyltransferase inhibitors AZA and DAC show antiproliferative and antimetabolic effects on T-ALL cells in vitro. Effects are further increased by combination with conventional cytostatics. Simultaneous application seem to be showing the strongest synergistic effects. Disclosure: No conflict of interest disclosed.

Abstracts

P850

GIMEMA ALL-rescue protocol as salvage regimen in relapsed/ refractory acute lymphoblastic leukemia/aggressive lymphoma: a single center retrospective analysis Nörenberg D.1, Arnold R.1, Flörcken A.1, Schmitt C.1, Dörken B.1, Westermann J.1 Medizinische Klinik m.S. Hämatologie, Onkologie und Tumorimmunologie, Charité Berlin – Universitätsmedizin Berlin, Berlin, Germany 1

Introduction: Relapsed or refractory acute lymphoblastic leukemia/ lymphoma remains a therapeutic challenge with allogeneic stem cell transplantation being the only option for long term survival. No salvage chemotherapy regimen has yet emerged as standard of care. Here we report on 18 heavily pretreated patients with relapsed/refractory acute lymphoblastic leukemia or aggressive lymphoma who received combination chemotherapy according to the GIMEMA ALL – rescue 97 protocol (3 g/m² HDAC d1-d5, 40 mg/m² idarubicine d3). Methods: In our database, we identified 18 patients that had received the GIMEMA rescue protocol: 14 patients with B-cell neoplasms (11 B-lineage ALL, including 2 Ph+ ALL, 2 Burkitt-lymphoma, 1 double hit DLBCL) and 4 patients with T-cell malignancies (3 T-ALL, 1 hepatosplenic TCL). 8/18 patients were in first relapse, 7 patients were primary or secondary refractory and 3 patients had relapsed after allogeneic SCT. In addition, 4 patients had CNS involvement. Patient records were analyzed for clinical characteristics, response to therapy, adverse events and subsequent allogeneic stem cell transplantation. Results: Median age of the patients was 34 (22-59) years with a mean Karnofsky performance status of 80 (60-100) %. On day 30-35, 61% (11/18) of the patients had achieved CR/CRi. 2 had a partial response, 4 had persistent or progressive disease and 1 patient died before response evaluation. Treatment-related mortality was 16% due to infectious complications (2 bacterial, 1 fungal sepsis). Other serious adverse events were manageable with no further ICU transferals. Notably, 66% of the patients were able to proceed to allogeneic stem cell transplantation (median time to transplant 9 weeks). Median progression-free survival (PFS) was 4 months, median overall survival (OS) 9 (1-60) months. Finally, 2/18 patients achieved long term survival (>4,5 years, >1,9 years), both had received allo-HSCT. Conclusion: The CR/CRi rate achieved by the GIMEMA ALL-rescue protocol was comparatively high. Therefore, the protocol can be considered a valuable option to induce remission before allo-HSCT. However, treatment-related toxicity/mortality is considerable and long-term survival was only observed in 2/18 patients. Thus, more effective and less toxic regimens are urgently needed. Immunotherapeutic strategies such as bispecific antibodies and adoptive T cell transfer offer interesting perspectives as a part of a multimodality therapy including allo-HSCT. Disclosure: No conflict of interest disclosed. P851

Plasmapheresis during acute liver failure after pegylated asparaginase in a patient with B-cell acute lymphoblastic leukemia Göpel W.1, Schnetzke U.1, Hochhaus A.1, Scholl S.1 Klinik für Innere Medizin II, Universitätsklinikum Jena, Hämatologie/ internistische Onkologie, Jena, Germany 1

Introduction: Treatment with asparaginase may induce apoptosis of acute lymphoblastic leukemia (ALL) cells by depletion of asparagine. The introduction of asparaginase has contributed to the improvement of the outcome of ALL patients. In children, long-term survival of ALL can be achieved in about 85% of patients. Half-life of asparaginase is significantly affected by additional modifications, e.g. pegylated asparaginase (PEG-asparaginase) has a prolonged half-life of about six days. Important side effects of asparaginase comprise hypersensitivity, pancreatitis, coagulopathy or liver dysfunction. Acute liver failure represents a rare complication following asparaginase treatment that is regularly associated with acute hepatitis.

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Methods: We report on a 44-year-old man with standard risk B-cell acute lymphoblastic leukemia (common ALL) who was treated according to the German ALL consensus protocol. Following induction chemotherapy phase I that was completed with the application of PEG-asparaginase (3750 IU absolute) our patient developed a functional acute liver failure with critical hyperbilirubinemia, hypoalbuminemia and especially a severe coagulopathy. Hyperbilirubinemia indicated a severe impairment of bilirubin secretion without primary evidence of cholestasis or acute hepatitis. Results: In this clinical constellation we initiated additional treatment with plasmapheresis that was performed for three consecutive days. Following plasmapheresis a continuous stabilization of fibrinogen and antithrombin III, an increase of cholinesterase and a slow decrease of bilirubin could be observed and was accompanied by a rapid improvement of clinical symptoms. The patient fully recovered from this severe complication after asparaginase treatment and achieved complete remission prior to phase II of induction chemotherapy. Conclusions: To our best knowledge, this is the first case of functional acute liver failure induced by PEG-asparaginase application. We hypothesise that plasmapheresis has contributed to the favorable outcome of this severe complication. Disclosure: Wibke Göpel: No conflict of interest disclosed. Sebastian Scholl: Expert Testimony: Förderprogramm Infektiologie der Fa. Gilead P852

Chromatin accessibility maps of chronic lymphocytic leukemia identify subtype-specific epigenome signatures and transcription regulatory networks Schmidl C.1, Rendeiro A.F.1, Strefford J.C.2, Walewska R.3, Davis Z.3, Farlik M.1, Oscier D.3, Bock C.1,4,5 CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria, 2University of Southampton, Faculty of Medicine, Southampton, United Kingdom, 3Royal Bournemouth Hospital, Department of Molecular Pathology, Bournemouth, United Kingdom, 4Medical University of Vienna, Department of Laboratory Medicine, Vienna, Austria, 5Max Planck Institute for Informatics, Saarbrücken, Germany 1

Introduction: Chronic lymphocytic leukemia (CLL) is characterized by substantial clinical heterogeneity, despite relatively few genetic alterations. This notion, and the recent discovery of recurring mutations in non-coding regulatory elements, chromatin remodelling factors, and CLL-subgroup-specific DNA methylation signatures suggest epigenetic contributions to the observed clinical heterogeneity. While prior studies of epigenome deregulation in primary cancer samples mostly focused on DNA methylation, recent technological advances make it possible to map chromatin landscapes in large patient cohorts. Most notably, the assay for transposase-accessible chromatin using sequencing (ATAC-seq) facilitates open chromatin mapping in scarce clinical samples, and ChIPmentation provides a streamlined, low-input workflow for genome-wide mapping of histone marks and transcription factors. Methods: To provide a basis for studying epigenome deregulation in CLL, we established genome-wide chromatin accessibility maps for 88 CLL samples from 55 patients using the ATAC-seq assay, and we also performed ChIPmentation and RNA-seq profiling for ten representative samples. Results: Based on the resulting dataset, we devised and applied a bioinformatic method that links chromatin profiles to clinical annotations. Our analysis identified sample-specific variation on top of a shared core of CLL regulatory regions. IGHV mutation status – which distinguishes the two major subtypes of CLL – was accurately predicted by the chromatin profiles, and gene regulatory networks inferred for IGHV-mutated vs. IGHV-unmutated samples identified characteristic differences between these two disease subtypes. Conclusion: We found widespread heterogeneity in the CLL chromatin landscape, we could associate distinct open chromatin patterns with clinical sample annotation and discover associated molecular and gene-regula-

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tory pathways. Finally, we established a community resource for studying epigenome deregulation in leukemia, and demonstrated the feasibility of chromatin accessibility mapping in cancer cohorts and clinical research. Disclosure: No conflict of interest disclosed. P853

Metabolic reprogramming of mesenchymals stem cells upon co-cultivation with primary CLL cells determined by mass spectrometry-based proteomics Schwarzmeier J.1, Mayer R.2, Slany A.1, Bileck A.2, Gerner C.2 K. Landsteiner Inst. f. Bioanalytische Chemie, Wien, Austria, 2Universität Wien, Abt. f. Analytische Chemie, Wien, Austria 1

Chronic lymphocytic leukemia (CLL) displays a strong dependency on tumor-stroma interactions. CLL cells were found to rapidly die when cultivated in vitro, whereas co-cultivation with mesenchymal stromal cells promotes cell proliferation and ensures leukemia cell survival. It is therefore of great interest to obtain a thorough picture of the exact molecular players involved in this critical interplay. While a number of protagonists are already well described, there are still unresolved issues concerning the effect of CLL cells on the metabolism of stromal cells. In this study, we co-cultivated for one week primary CLL cells from peripheral blood with mesenchymal stem cells (hMSC). Subsequently, hMSC were lysed and fractionated to obtain cytoplasmic, nuclear and secreted proteins. To allow comparative analyses B-cells from healthy individuals and hMSC were co-cultivated. Mass spectrometry-based proteomic analysis resulted in the identification and relative quantification of more than 3700 proteins. Comparative analyses between co-cultures with CLL and healthy B-cells showed that already at a ratio of 0.25% CLL cells could induce a significant proteomic alterations in hMSC. Furthermore, hMSC were found to undergo a metabolic change upon co-cultivation with CLL cells, an effect which was most pronounced in the secreted protein fraction. Annotation analyses according to gene ontology revealed that proteins related to “generation of precursor metabolites” were significantly up-regulated in hMSC upon co-culture with CLL cells. This indicates that these stromal cells “feed” the leukemia cells by producing and secreting high energy metabolites, satisfying the elevated metabolic demands of the CLL cells. Revealing the metabolic interactions between CLL cells and surrounding stromal cells by mass spectrometry-based methods should contribute to the development of innovative treatment strategies. Disclosure: No conflict of interest disclosed. P854

The effects of CDK-Multikinase inhibition in Chronic Lymphocytic Leukemia on proliferation and survival: In vitro characterization of involved pathways Leiser L.1, Ölsner M.1, Bogner C.1 Klinikum rechts der Isar der Technischen Universität München, Hämatologie/ Onkologie, München, Germany 1

Introduction: Despite emerging novel therapeutic options, up to now, there is still no cure for the majority of patients with Chronic Lymphocytic Leukemia, the most common leukemia in the western world. For the last years many anti tumor compounds have been developed including cyclin-dependent kinase (CDK)-Inhibitors. CDK inhibition plays an important role in interfering with cell cycle and programmed cell death, key hallmarks of cancer. Methods: We incubated primary cells of CLL patients with the multiple CDK target inhibitor (Bay 1000394) in vitro and analyzed proliferation and survival by flow cytometry and Western blotting in resting and stimulated CLL cells as well as in CLL cells in contact with supportive stroma. Results: Bay 1000394 induced distinct programmed cell death in resting and proliferating CLL cells at early time points at low nanomolar concentrations. In proliferating CLL cells cell cycle progression was inhibited.

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The high rate of cell death we observed was accompanied by phosphatidylserin exposure on the outer cell membrane, loss of mitochondrial membrane potential as well as the detection of DNA strand breaks. Caspase 8, Caspase 9 and 3 activation reflect intrinsic and extrinsic apoptosis induction. We observed the activation of proapoptotic Bcl-2 family members Bid, Bim and Bax, and down regulation of several anti-apoptotic proteins. A broad-spectrum caspase inhibitor (Z-VAD-FMK) was only in part able to inhibit cell death, suggesting that there are other pathways involved distinct from apoptosis. Conclusion: The multikinaseinhibitor Bay 1000394 shows potent activity in B-CLL cells in vitro and may add to the repertory of targeted therapies that should overcome the need of classic chemotherapy in this still challenging disease. Disclosure: No conflict of interest disclosed. P855

Three years after the start of enrolment: Current data from the prospective CLL-registry of the German CLL Study Group Fink A.M.1, Maurer C.1, Bahlo J.1, Kluth S.1, Linde H.2, Aldaoud A.3, Illmer T.4, Dengler J.5, Klausmann M.6, Lathan B.7, Al-Sawaf O.1, Fischer K.1, Wendtner C.-M.8, Eichhorst B.1,9, Hallek M.1,9 Uniklinik Köln, Centrum für Integrierte Onkologie Köln Bonn, Klinik 1 für Innere Medizin, Köln, Germany, 2Medizinisches Versorgungszentrum für Blut- und Krebserkrankungen, Potsdam, Germany, 3Gemeinschaftspraxis für Hämatologie und Onkologie, Leipzig, Germany, 4Gemeinschaftspraxis Hämatologie-Onkologie, Dresden, Germany, 5Onkologische Schwerpunktpraxis Heilbronn, Heilbronn, Germany, 6Gemeinschaftspraxis Hämatologie/ Onkologie, Aschaffenburg, Germany, 7Gemeinschaftspraxis für Hämatologie und Onkologie, Dortmund, Germany, 8Klinikum Schwabing, Department of Haematology, Oncology, Immunology, Palliative Care, Infectious Diseases and Tropical Medicine, München, Germany, 9Cluster of Excellence in Cellular Stress Responses in Aging-associated Diseases, Cologne, Germany, Köln, Germany 1

Introduction: The registry of the German CLL-Study Group (GCLLSG) was established to collect data from patients (pts) in and outside of clinical trials. Methods: Pts with confirmed diagnosis of CLL or related entities (B-PLL, T-PLL, SLL, T/NK-LGL, HCL or Richter’s transformation) were included since September 2013. Comprehensive information on the disease status (stage, molecular genetics and cytogenetics, treatments, responses, secondary malignancies, quality of life and patients’ compliance were collected. Results: As of May 2016 a total of 2258 pts were enrolled in the registry. 85% (1926) of pts did not participate in any of the GCLLSG studies before. The vast majority of the pts were diagnosed with CLL (2139). At the time point of enrollment 64% (1467) pts were treatment-naïve, whereas 36% (791 pts) had at least received a median number of 1 treatment ( range 1-13). 650 CLL-pts enrolled until October 2015 were analysed. 62% (403) of the pts were male, median age at registration was 72 years (range 38-92). Median observation time between initial diagnosis and last survival visit was 103.4 months (range 4.4-459.4) and 61.4 months (range 0.0-340.9) between start of first treatment and last survival visits. At time point of initial diagnosis 57% had Binet stage A, 29% Binet B and 14% Binet C. 94% of the pts had a CIRS score lower than 6 (median 2 points (range 0-12). 25%/159 pts were treated with fludarabine, cyclophosphamide and rituximab (FCR) and 33%/214 pts were treated with bendamustine and rituximab (BR) in the frontline setting, less frequent, pts received chlorambucil (11%/71); FC (9%/56) and bendamustine mono (5%/35). Also, a wide variety of other treatment approaches was documented. Complete Remission (CR) or complete remission with incomplete bone marrow recovery (CRi) was reported for FCR with 28,5% and 3,3% respectively. Results for BR were comparable with 25,1% CR and 1,5% CRi respectively. The rate for non-responders was 1,3% for FCR and 3,1% for BR. Response was unknown for 5% of pts treated with FCR and 8,9% treated with BR. So far, 43 cases of death have been reported, 16 due to progressive disease, infections (14), concomitant diseases (7) and other malignancies (4), cause of death was unknown in 6 cases.

Abstracts

Conclusions: More than 2250 pts were enrolled in the DCLLSG-registry so far. An updated data set including Kaplan Meier curves for survival will be presented at the meeting. Disclosure: Anna Fink: Financing of Scientific Research: ja; Expert Testimony: ja Michael Hallek: Advisory Role: ja; Financing of Scientific Research: ja; Expert Testimony: ja P856

CLL – patient registry (cumulative dose of rituximab) Linde H.1, Blumenstengel K.2, Göhler T.3, Spohn C.4, Göttel R.5, Tessen H.W.6, Projektgruppe Internistische Onkologie (PIO) Medizinisches Versorgungszentrum für Blut- und Krebserkrankungen, Potsdam, Germany, 2Onkologische Schwerpunktpraxis, Eisenach, Germany, 3 Onkologische Schwerpunktpraxis, Dresden, Germany, 4Onkologische Schwerpunktpraxis, Halle, Germany, 5rgb Onkologisches Management GmbH, Sarstedt, Germany, 6Onkologische Kooperation Harz, Goslar, Germany 1

Introduction: The chronic lymphocytic leukaemia (CLL) is the most common leukemic disease in Central Europe. The median age of onset is between 70 and 75 years. The combination of bendamustine and rituximab has proven effective for the treatment of this disease in clinical trials and everyday use. Does the cumulative dose of rituximab influence the response rates and the survival? Methods: Since 2008, 61 hemato-oncological practices from 16 federal states within the project team of internal oncology (PIO) have been documenting disease histories of patients with a chronic lymphocytic leukaemia in the registry ONCOReg. 812 patients received a bendamustine-based therapy, 569 (70.1%) as first-line therapy, 127 (22.3%) of them bendamustine mono and 442 (77.7%) in combination with rituximab. Results: Tab. 1. Patients`characteristics

n = 442 Number Gender (m/f in %) Median age Without B-symptoms ECOG 0/1/2/3 (%) BINET A/B/C (%) Median number of cycles Average dose of bendamustine Average dose of rituximab

Cumulative dose of rituximab <2875 mg/m² 274 (62.0%) 61.2/38.8 73 (42–92) years 173 (63.1%) 28.1/58.0/13.5/0.4 13.1/53.3/33.6

Cumulative dose of rituximab ≥2875 mg/m² 168 (38.0%) 67.9/32.1 70 (36–84) years 102 (60.7%) 28.6/56.5/14.9/0 7.7/56.5/35.7

5 (1–8)

6 (3–8)

700 mg/m²

973 mg/m²

1,624 mg/m²

2,894 mg/m²

Response: The objective remission rate is at 88.9% (47.1% CR/41.8% PR). Given a cumulative dose of rituximab of ≥2,875 mg/m², the rate of complete remissions was at 60.7% compared to 38.7% (rituximab < 2,875 mg/ m²). The highest CR rate was reached with BRP and a cumulative dose of rituximab ≥2,875 mg/m² (64.9%). Survival: The progression-free survival of the overall population is at 48.0 months at median and differs depending on the cumulative dose of rituximab: 44.5 months < 2,875 mg/m²; 50.5 months ≥2,875 mg/m². After a median follow-up of 30.4 months, the median overall survival of the overall population has not been reached yet. The 3-year survival rate is at 84% and has reached 90% with patients who received ≥2875 mg/m² of rituximab (compared to 81% < 2875 mg/m² of rituximab). Conclusion: Response rate, progression free and overall survival depend on the cumulative dose of rituximab and improve when at least at 2,875 mg/m² Further data will be presented.

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Disclosure: Hartmut Linde: Financing of Scientific Research: Roche, Mundipharma; Other Financial Relationships: Fortbildungskosten durch Mundipharma Hans-Werner Tessen: No conflict of interest disclosed. P857

Survival improvement of patients with chronic lymphocytic leukaemia (CLL) in routine care. A retrospective analysis of all patients with CLL who were treated in an oncology group practice in Germany between 1995-2015 Weide R.1, Feiten S.2, Chakupurakal G.1, Friesenhahn V.2, Kleboth K.2, Köppler H.1, Lutschkin J.2, Thomalla J.1, van Roye C.1, Heymanns J.1 Praxisklinik für Hämatologie und Onkologie, Koblenz, Germany, 2Institut für Versorgungsforschung in der Onkologie, Koblenz, Germany 1

Introduction: Progress has been made in the diagnosis and treatment of patients with CLL who receive their treatment within prospective clinical trials. Clinical trials only include patients who fulfill the selection criteria and hence do not reflect outcomes in routine practice. Data with regards to the diagnosis, treatment and survival of patients in routine care reflecting the improvements in survival following implementation of newer therapy strategies and agents are not available. Methods: A retrospective analysis of all patients with CLL who were treated in an oncology group practice in Germany between 1995-2015. Relevant clinical data concerning diagnosis, treatment and survival were transferred from clinical files into a database and analysed statistically using SPSS. Results: 676 CLL patients with a median age of 67 (35-92) were identified. At initial diagnosis 521 patients (77.1%) were in Binet stage A, 83 (12.3%) Binet stage B and 36 (5.3%) Binet stage C. Due to external diagnosis of 36 patients (5.3%) the stage at initial diagnosis couldn’t be retrieved. 361 patients (53.4%) never received any treatment. 315 patients (46.6%) needed therapy with a median of 2 therapy lines (1-13). Regimens most frequently applied were: Bendamustine-containing (68.9%), Rituximab-containing (65.4%), Chlorambucil-containing (55.9%), Bendamustine+Rituximab combinations (48.3%) and Fludarabine-containing (39.4%). Regimens containing recently approved drugs could be found as well. 5.1% received Ibrutinib, 2.9% Obinutuzumab and 2.2% Idelalisib. 17.8% of patients were treated within a clinical trial. 5 and 10 year overall survival was 84.5% and 60.4%. Median overall survival according to Binet stage was 13 years for Binet A, 9 years for Binet B and 9 years for Binet C. Patients who needed therapy had a median overall survival of 11 years (0-42) compared to 17 years (0-27) of patients who never needed any therapy. Conclusions: 53.4% of CLL-patients never needed any therapy. Patients who needed therapy had a much lower life expectancy compared to patients who never needed therapy. Treatment consisted mainly of Bendamustine, Rituximab, Chlorambucil, Bendamustine+Rituximab combinations and Fludarabine leading to a marked prolongation of survival compared to historical controls and registry data. Disclosure: No conflict of interest disclosed. P858

Autoimmune myelitis in a CLL patient undergoing treatment with ibrutinib Wanner D.1, Bohn J.-P.1, Rudzki J.1, Stockhammer G.2, Steurer M.1 Universitätsklinik Innsbruck, Innere Medizin V, Innsbruck, Austria, Universitätsklinik Innsbruck, Universitätsklinik für Neurologie, Innsbruck, Austria 1 2

Background: Myelitis is a neurological disorder caused by inflammation of the spinal cord, mostly in association with infectious or autoimmune diseases. Symptoms, including paraesthesia and autonomic dysfunction, develop within hours/days and immediate therapy is required. Association with CLL has not been described yet. Case Presentation: A 44-year-old man was diagnosed with CLL 12 years ago. Previous therapies consisted of R-CHOP, FCR and R-Bendamustine

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(+ NOX-A-12). However, only two months after 3rd-line therapy he required further treatment. Of note, the patient has never had any neurological symptoms, CLL associated autoimmune phenomena or other autoimmune diseases before. Ibrutinib was started and the patient responded immediately in terms of vanishing B-symptoms and regression of massive lymphadenopathy. After one month the patient complained of paraesthesia in the left foot spreading within hours to the right leg, the pelvic floor, the genital area and onset of a voiding dysfunction. CT, CSF, infectiological and microbiological examinations were without any pathological findings. MRI revealed signal enhancements of the spinal cord at the thoracolumbar transition, at TH7 and several intracranial subcortical lesions. Based on these findings and the clinical presentation, the patient was diagnosed with an autoimmune myelitis. No other autoimmune symptoms were detectable. Treatment with methylprednisolone was induced and ibrutinib was continued due to ultra-high-risk CLL. Two-month follow-up showed persistent paraesthesia, though the voiding dysfunction vanished completely. 10-months follow-up revealed further improvement of the clinical symptoms, only a hypaesthesia of the pelvic floor and the genital area was still present. MRI showed a regression of enhancements. Ibrutinib was otherwise well tolerated and the patient achieved an excellent and ongoing partial remission under continuous ibrutinib treatment. Conclusion: The role of ibrutinib in the setting of CLL-associated autoimmunity needs to be clarified as both positive and negative effects on autoimmune phenomena have been reported. To the best of our knowledge autoimmune myelitis has never before been reported in CLL. Thus, our case suggests, that it might be an autoimmune adverse event, possibly triggered by ibrutinib. Future investigations and post-marketing surveillance are necessary to provide a more detailed safety profile of ibrutinib including its immunomodulating properties. Disclosure: David Wanner: No conflict of interest disclosed. Michael Steurer: Advisory Role: Janssen-Cilag; Financing of Scientific Research: Janssen-Cilag; Expert Testimony: Janssen-Cilag P859

Bendamustine triggered paraneoplastic pemphigus in a patient with chronic lymphocytic leukemia Föhring D.1, Dührsen U.1, Hammers C.M.2, Dürig J.1 Klinik für Hämatologie, Universitätsklinikum Essen, Universität DuisburgEssen, Essen, Germany, 2Universität zu Lübeck, Klinik für Dermatologie, Lübeck, Germany 1

Background: Paraneoplastic pemphigus (PNP) is a rare and critical mucocutaneous autoimmune disorder characterized by painful mucosal eruptions. It is generally associated with lymphoproliferative diseases including chronic B-cell lymphocytic leukemia. Patients with PNP often show serum autoantibodies against various cell adhesion proteins such as plakins and/or cadherins. The primary cause of PNP is unknown in most cases, it could however be triggered by specific drugs used in lymphoma treatment. Treatment of PNP remains difficult because of its resistance to conventional therapies. Patient / Methods: Here we report about an 64-year-old male patient with CLL being diagnosed in January 2013. He required therapy two years later in January 2015 and was treated with rituximab/bendamustine (R-B). After the second cycle he suffered from severe painful mucosal erosive lesions that could not be influenced by antibiotics or aciclovir. Furthermore he developed skin involvement with fragile blisters. We detected serum autoantibodies against envoplakin and desmoglein, compatible with the diagnosis of a PNP. Interestingly the autoantibodies were not detectable in serum samples collected prior to the initiation of R-B therapy. Immunsuppressive treatment with rituximab, cyclophosphamide and dexamethasone was administered for six cycles which resulted in partial remission of CLL but unfortunately no improvement of PNP lesion was observed. Due to PNP the general condition of the patient and his quality of life continued to deteriorate. Thus we performed protein A-immunoadsorption therapy in combination with rituximab and intravenous dexamethasone pulse therapy. As a result mucosal eruptions were alleviated and weight

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gain of 10 kg was achieved. Serological follow up revealed a decrease of autoantibodies for some tissue antigens and complete disappearance for other antigens targeted in PNP Results / Conclusions: Here we present the first case of bendamustine triggered PNP in a B-CLL patient. This case also emphasizes the difficult treatment and shows that immunoadsorption therapy can be effective in patients with an incomplete response to immunsuppressives alone. Disclosure: No conflict of interest disclosed. P860

Profit center analysis of chronic lymphatic leukemia cases: a hospital management perspective Kron F.1, Nadine K.1, Kostenko A.1, Dohle I.2, Müller D.3, Bahlo J.1, Bergwelt M.1, Eichhorst B.1, Hallek M.1, Zander T.1 Uniklinik Köln, Klinik I für Innere Medizin, Köln, Germany, 2Uniklinik Köln, Controlling, Köln, Germany, 3Uniklinik Köln Institut für Gesundheitsökonomie und Klinische Epidemiologie, Köln, Germany 1

Objectives: Chronic lymphocytic leukemia (CLL) is a disease specially afflicted by demographic and treatment changes. Here we analyzed economic risks in hospitalized CLL patients from a management perspective in order to support strategic controlling. Methods: 112 CLL patients hospitalized in 2013 and 2014 at the university hospital of cologne were analyzed. To assess profit margins (PM) per case (PC), Diagnosis Related Groups (DRG) reimbursement data were merged with an internal DRG cost accounting scheme based on the Institute for the Hospital Remuneration System (InEK) depending on age, prognostic factors and DRG key performance indicators. Results: 294 patient cases coded by 19 different DRG with strongly fluctuating cost-revenue ratios were found with an overall negative profit margin of € -137,147 (total €2.9 million) .The DRG R61H (`lymphoma and non-acute leukemia`) was identified as the most frequented case category (174 cases, 61.3%) with a mean deficit PC of € -814. Subanalysis of these cases demonstrated, the payments were not cost-covering primary caused by drug costs and length of stay. Statistical significant differences in PM per DRG case concerning age at treatment, length of stay and number OPS codes (p < 0.05) were found, not, however, with regard to prognostic factors and comorbidities. Conclusion: Treatment of CLL patient in a research driven tertiary care hospital is not cost-effective. This observation demonstrate the need for novel care and or reimbursement structures in research-driven CLL treatment, to allow for cost covering for health care providers. Effective cost-revenue controlling is crucial to avoid mayor economic risk from a hospital management perspective.

ond line starting in 2015 in comparison to 2012. Therefore, data of 118 first line and 32 second line therapies in 2015 and in contrast, 81 first line and 33 second line therapies in 2012 were included. Results: 199 B-CLL patients in first line (2015: 118 patients, 2012: 81 patients) were included in the statistical analysis. The median age of both groups was nearly identical (2015: 74 years, 2012: 73 years). In contrast, there was a slight difference in the performance status ECOG (2015: 83.1%, 2012: 74.1% with ECOG 0) which is not statistically significant. Patients in second line therapy in 2015 were in median apparently older than second line patients in 2012 (2015: 78 years, 2012: 71 years). Also the performance status ECOG of patients in 2012 was marginally better than of those in 2015 (2015: 59.4%, 2012: 75.8% with ECOG 0). In the analysis, an obvious reduction of former frequently used therapies was seen from 2012 to 2015 in both therapy lines. Exemplary the use of Chlorambucil single therapies in first line diminished from 12.4% in 2012 to 2.5% in 2015 and of R-CHOP in second line from 12.1% in 2012 to 3.1% in 2015. On the other hand, the portions of new therapy regimen like Idelalisib+Rituximab in first line or Ibrutinib in second line increased from 2012 to 2015 in a rate of 7.6% in the first line and 21.9% in the second line. Conclusions: The results of this analysis showed a change of the prescriptions of former frequently used therapies to regimen with new drugs from 2012 to 2015. The changes concerned mostly older regimen like Chlorambucil single or R-CHOP whereas other established therapies like R-Bendamustine or R-FC were prescribed in comparable frequencies. Disclosure: No conflict of interest disclosed.

Posterdiskussion

Gastrointestinale Tumoren, Hepatozelluläres Karzinom, Pankreaskarzinom P862

CXCR4, CXCR7 and CXCL12 expression is not a predictive factor in patients with resected pancreatic cancer – results from the CONKO-001 trial Striefler J.K.1, Wislocka L.1, Sinn M.1, Denkert C.2, Juehling A.1, Pelzer U.1, Bischoff S.3, Bahra M.4, Oettle H.1, Bläker H.2, Riess H.1, Lohneis P.2 Charité Universitätsmedizin Berlin, CC14 Med. Klinik m. S. Hämatologie, Onkologie und Tumorimmunologie, Berlin, Germany, 2Charité Universitätmedizin Berlin, Campus Mitte, Institut für Pathologie, Berlin, Germany, 3Charité Universitätsmedizin Berlin, CONKO Studiengruppe, Berlin, Germany, 4Charité Universitätsmedizin Berlin, Klinik für Allgemein-, Viszeralund Transplantationschirurgie, Berlin, Germany 1

Introduction: B-CLL is one of the most occuring hematological diseases and the pharmaceutical development of new drugs in recent years resulted in a broad range of new therapy regimen. This retrospective, observational study focused on a comparison and an analysis of possible changes in the prescriptions of regimen from 2012 to 2015. Methods: Data about the used therapy regimen in B-CLL were collected under real-world conditions in 26 German oncological practices. For this analysis, it was determined to regard the therapy regimen of first and sec-

Introduction: Pancreatic cancer is a disease with a dismal prognosis. Several biomarkers have been tested in heterogeneous patient groups. Our aim was to evaluate the expression of the chemokine receptors CXCR4, CXCR7 and their common ligand CXCL12 as predictive and/or prognostic factors of adjuvant chemotherapy with gemcitabine and to evaluate their association with type of metastases. Methods: CXCR4, CXCR7 and CXCL12 expression was analyzed in resected pancreatic cancer tissues by immunohistochemistry from 160 of 368 patients enrolled in the CONKO-001 trial, which established gemcitabine as a standard of adjuvant therapy. The correlation of CXCR4/ CXCR7/CXCL12 expression with survival was analyzed by fitting a Cox proportional hazards model with respect to the type of recurrence (local and/or distant) and the location of metastases. Kaplan-Meier analyses for median disease-free survival (DFS) and overall survival (OS) were performed in dependence on CXCR4, CXCR7, and CXCL12 expression. Results: CXCR4 was positive in 128/160 patients (80%), CXCR7 in 63/157 (40%) and CXCL12 in 110/158 patients (70%). There was no correlation of CXCR4/CXCR7/CXCL12 expression with type of recurrence (local and/ or distant) or the location of metastases. Expression of CXCR4 (median DFS low vs. high 10.3 vs 11.0, p = 0.31), CXCR7 (median DFS low vs. high 11.2 vs 9.5, p = 0.87), and CXCL12 (median DFS low vs. high 9.0 vs 11.8, p = 0.89) did not show a difference in DFS and OS neither in the overall

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–294

Disclosure: No conflict of interest disclosed. P861

Distribution of therapy regimen in patients with B-cell chronic lymphocytic leukemia (B-CLL) in 2015 in comparison to treatment strategies in 2012 based on real-world-evidence data of German oncological practices Feuerbach M.1, Freigang F.1, Schulte C.2, Hurtz H.-J.3, Schwarzer A.4, Lipp R.1 GermanOncology GmbH, Hamburg, Germany, 2Gemeinschaftspraxis Hämatologie-Onkologie, Dortmund, Germany, 3Gemeinschaftspraxis und Tagesklinik, Halle (Saale), Germany, 4Gemeinschaftspraxis für Hämatologie und Onkologie, Leipzig, Germany 1

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study population nor in the stratified analysis according to treatment arm. The multivariable analysis including covariates treatment arm, resection status, grading, Karnofsky performance status, nodal involvement, tumor stage, and expression of CXCR4 did not show a significant correlation with disease-free survival (HR 0.92, CI 0.62-1.36). Conclusion: Our data show neither a predictive nor a prognostic role of CXCR4, CXCR7, and CXCL12 in patients with pancreatic cancer. In contrast to previously reported studies, our results do not confirm an association between the expression of CXCR4, CXCR7, and their common ligand CXCL12 with the type of recurrence. Nevertheless, further studies are needed to validate their role in predicting recurrence patterns. Disclosure: No conflict of interest disclosed. P863

Effects of nanoliposomal irinotecan (nal-IRI;MM-398) ± 5-Fluorouracil und leucavorin (5-FU/LV) on quality of life (QoL) in NAPOLI-1: a phase 3 study in patients (pts) with metastatic pancreatic adenocarcinoma (mPAC) previously treated with gemcitabine-based therapy Siveke J.T.1, Cubillo A.2, Blanc J.-F.3, Melisi D.4, Von Hoff D.D.5, WangGillam A.6, Chen L.-T.7, Becker C.8, Mamlouk K.8, Belanger B.8, Yang Y.9, de Jong F.10, Hubner R.11 West German Cancer Center, University Hospital Essen, Essen, Germany, Clara Campal-Hospital, Madrid, Spain, 3Hôpital Saint-André, Bordeaux, France, University of Verona, Verona, Italy, 5Virgina G. Piper Cancer Center, Scottsdale, United States, 6Siteman Cancer Center, Washington University, Saint Louis, United States, 7NHRI, National Institute of Cancer Research, Taipei, Taiwan, Republic of China, 8Merrimack Pharmaceuticals, Inc., Cambridge, United States, 9 Baxalta US Inc, Cambridge, United States, 10Baxalta GmbH, Zϋrich, Switzerland, 11 Christie Hospital NHS Foundation Trust, Manchester, United Kingdom 1 2 4

Introduction: Here we report QoL results from the NAPOLI-1 trial: a randomized phase 3 study of nal-IRI plus 5-FU/LV vs 5FU/LV in pts with mPDAC previously treated with gemcitabine-based therapy. Methods: QoL was assessed using the EORTC-QLQ-C30, which includes functional and symptom scales, and a global health and QoL scale. Pts completed the questionnaire at treatment start, every 6 weeks (wks), and 30 days post-follow-up visit. Pts who provided baseline and ≥1 subsequent assessment were included. Linear transformations were applied to raw scores to produce reported scores in the 0-100 range. Pts were classified as improved (≥10% increase in scale of breadth at a post-baseline time point and remained above baseline for ≥6 wks), worsened (did not meet improvement criteria and died, or had ≥10% decrease from baseline in scale of breadth at a post-baseline time point), or stable (did not meet criteria for improvement or worsening) for each subscale. Pairwise treatment group comparisons on response classification were performed for each subscale and adjusted for multiplicity to control false discovery rate at 0.05 level for the 15 comparisons. Results: 154 pts were included in the analysis; 69% (49/71) in the nalIRI+5-FU/LV group and 53% (44/83) in the 5-FU/LV group had evaluable data at 12 wks. At baseline, median Global Health Status scores were near the midpoint of the scoring range, median Functional Scale scores were high, and Symptom Scale scores were low, with baseline values similar between groups. The observed median change in score at 12 wks was 0 for both treatment groups for Global Health Status and for the following subscale scores: role functioning, emotional functioning, cognitive functioning, social functioning, nausea and vomiting, pain, dyspnea, insomnia, appetite loss, constipation, diarrhoea, and financial difficulties. For subscale scores where the median change was not 0 (nal-IRI+5-FU/LV: physical functioning and fatigue), the between-group differences were not substantial. There were no significant differences in the proportion of pts classified as improved, worsened, or stable between the treatment groups. Across subscales, adjusted P values for the comparisons were >0.05 (NS). Conclusion: While limited by pt numbers, in this analysis nal-IRI+5-FU/ LV-treated pts tended to maintain baseline QoL over 12 wks. There were

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no significant differences versus 5-FU/LV-treated pts in QoL response despite the addition of a second cytotoxic agent Disclosure: Jens T. Siveke: Advisory Role: Baxalta adivsory boards Richard Hubner: Advisory Role: Baxalta advisory boards P864

Nab-paclitaxel/Gemcitabine first line therapy in patients with metastatic pancreatic carcinoma and high-bilirubin values – data from the German QoliXane pancreatic cancer registry zur Hausen G.1, Waidmann O.2, Woerns M.-A.3, Höffkes H.-G.4, Doerfel S.5, Zahn M.-O.6, Aldaoud A.7, Stauch M.8, Springfeld C.9, Haertel N.10, Reichart A.1, Götze T.-O.1, Schwarz S.1, Pauligk C.1, Römmler-Zehrer J.11, Hofheinz R.-D.12, Al-Batran S.-E.1 Institute of Clinical Cancer Research, Krankenhaus Nordwest gGmbH, Frankfurt am Main, Germany, 2University Hospital Frankfurt, Medical Hospital 1, Frankfurt am Main, Germany, 3University Hospital Mainz, 1. Medical Hospital, Mainz, Germany, 4MVZ Osthessen, Fulda, Germany, 5Onkozentrum Dresden/ Freiberg, Dresden, Germany, 6MVZ Onkologische Kooperation Harz, Goslar, Germany, 7Forschungsgesellschaft Dr. Aldaoud/Dr. Schwarzer, Leipzig, Germany, 8 Practice Dr. Stauch, Kronach, Germany, 9NCT Heidelberg, Heidelberg, Germany, 10 University Medicine Mannheim, II. Medical Hospital, Mannheim, Germany, 11 Celgene GmbH, München, Germany, 12University Medicine Mannheim, III. Medical Hospital, Mannheim, Germany 1

Introduction: Hyperbilirubinaemia is a common disease effect in patients (pts) with metastatic pancreatic cancer (mPC). As clinical trials often exclude them, data on management of these pts are rare. In the framework of a German observational multicenter study (QoliXane), quality of life and therapy data are currently being collected in pts with mPC receiving a combination of nab-paclitaxel and gemcitabine. This is an interim analysis on hyperbilirubinaemia management. Methods: Pts were included to this analysis if they entered the trial with a bilirubin level ≥ 1.2 mg/dl and completed at least 2 cycles. Bilirubin levels were documented for up to 4 cycles and methods of hyperbilirubinaemia management have been assessed. A both descriptive and explorative analysis was performed using IBM SPSS V 23. Results: 25 of 294 pts (8.5%) were included. Mean bilirubin level was 2.96 mg/dl (range 1.2-12.3) at baseline and dropped considerably by the 2nd cycle to 0.84 (range 0.29-3.9; p = 0.0001). Bilirubin levels decreased in 24 (96%) and increased in 1 (4%) pts upon treatment start. 18 (72%) pts started treatment with standard dosage, 7 (28%) with a reduced regime. 10 (40%) pts underwent additional intervention: either stenting (7 pts, 28%) or bile duct anastomosis (3 pts, 12%). Mean bilirubin values dropped from 4.59 to 1.09 in pts with and from 1.87 to 0.68 in pts without additional intervention. Grade 3/4 toxicity was seen in 60% of pts and most common 3/4 events were anemia, nausea, and fever. Tab. 1. Mean Bilirubin Levels (1)

All pts (n = 25) no add. intervention (n = 15) any add. intervention (n = 10) add. stenting (n = 7) add. bile duct anastomosis (n = 3) p (all pts)(2)

Baseline 2.96 1.87 4.59 4.52 4.80

Cycle 2 0.84 0.68 1.09 1.19 0.87 0.0001

Cycle 3 0.83 0.92 0.68 0.84 0.47 0.002

Cycle 4 0.53 0.61 0.46 0.35 0.53 0.008

(1) n´s indicate # of pts at baseline (2) Wilcoxon test to baseline (related samples), level of significance p = 0.05

Conclusions: Data show that bilirubin levels drop considerably after start of nab-pac/gem therapy. The treatment seems to be feasible, although considerable frequencies of Grade 3/4 toxicities were observed. Disclosure: Gerrit zur Hausen: Other Financial Relationships: Reisekosten: Celgene, Lilly Salah-Eddin Al-Batran: Advisory Role: Merck, Roche, Celgene, Lilly, Nordic Pharma; Financing of Scientific Research: Merck, Roche, Celgene, Lilly, Nordic Pharma; Expert Testimony: Merck, Roche, Celgene, Vifor, Medac, Hospira, Lilly

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P865

P866

Quality-adjusted time without symptoms or toxicity (Q-TWiST) of nanoliposomal irinotecan (nal-IRI;MM-398) plus 5-fluorouracil and leucavorin (5-FU/LV) vs 5-FU/LV alone in patients with metastatic pancreatic adenocarcinoma (mPAC) previously treated with gemcitabine-based therapy

Nabpaclitaxel plus gemcitabine in patients with advanced pancreatic cancer suffering hyperbilirubenemia

Pelzer U. , Blanc J.-F. , Melisi D. , Cubillo A. , Von Hoff D.D. , WangGillam A.6, Chen L.-T.7, Siveke J.T.8, Wan Y.9, Solem C.T.9, Botteman M.9, Yang Y.10, de Jong F.11, Hubner R.12 1

Charite-Unversitätsmedizin, Berlin, Germany, 2Hôpital Saint-André, Bordeaux, France, 3University of Verona, Verona, Italy, 4Clara Campal-Hospital, Madrid, Spain, 5Virgina G. Piper Cancer Center, Scottsdale, United States, 6Siteman Cancer Center, Washington University, Saint Louis, United States, 7NHRI, National Institute of Cancer Research, Taipei, Taiwan, Republic of China, 8West German Cancer Center, University Hospital Essen, Essen, Germany, 9Pharmerit International, Bethesda, United States, 10Baxalta US Inc, Cambridge, United States, 11Baxalta GmbH, Zϋrich, Switzerland, 12Christie Hospital NHS Foundation Trust, Manchester, United Kingdom 1

Introduction: In the primary analysis of the phase 3 NAPOLI-1 trial, nalIRI+5-FU/LV significantly improved median overall survival (OS; 6.1 vs 4.2 months; hazard ratio [HR]=0.67; P = 0.012) and progression-free survival (PFS; 3.1 vs 1.5 months; HR = 0.56; P = 0.0001) vs 5-FU/LV alone in mPAC patients previously treated with gemcitabine-based therapy. Here we report between-treatment differences in quality-adjusted survival using the Q-TWiST methodology, commonly used in oncology. Methods: The total 12-month survival in NAPOLI-1 intent-to-treat cohort was partitioned into time before disease progression without toxicity grade ≥3 (TWiST), time with adverse event grade ≥3 (TOX), and time of disease progression (REL). Mean Q-TWiST was calculated by multiplying mean time spent in each health state by its respective utility. In the base case, the utility for TWiST (uTWiST), toxicity (uTOX), and post-progression (uREL) were set to 1.0, 0.5, and 0.5, respectively. The relative gain in Q-TWiST in nal-IRI+5-FU/LV over 5-FU/LV was calculated as the difference in Q-TWiST divided by the OS of the 5-FU/LV group. Non-parametric bootstrapped 95% confidence intervals (CIs) were derived around estimates. Threshold analyses varied uTOX and uREL between 0.0 and 1.0. An additional scenario analysis was conducted using the per protocol (PP) population. Results: Compared with patients receiving 5-FU/LV (n = 119), those receiving nal-IRI+5-FU/LV (n = 117) spent significantly more time in TWiST (mean 3.4 vs 2.4 months) and TOX (1.0 vs 0.3 months), but similar time in REL (2.5 vs 2.7 months). After weighing time spent in TWiST, TOX, and REL with their respective base-case utilities, nal-IRI+5-FU/LV resulted in 1.3 months (95% CI: 0.4-2.1; 5.1 vs 3.9) greater Q-TWiST, with relative Q-TWiST gain of 24%. In the threshold analyses varying uTOX and uREL, the Q-TWiST ranged from 0.9 to 1.7 months, and the relative Q-TWiST gain ranged from 17% to 31%. In the PP population, Q-TWiST was also significantly superior in patients recieving nal-IRI+5-FU/LV (Q-TWiST gain = 1.8 months; 95% CI: 0.7-3.0). Conclusion: In NAPOLI-1, nal-IRI+5-FU/LV resulted in statistically significantly and clinically important gains in quality-adjusted survival vs 5-FU/LV alone. This confirms the clinical outcome benefit of nal-IRI+5FU/LV in patients with mPAC. Disclosure: Uwe Pelzer: Advisory Role: Baxalta adivsory boards Richard Hubner: Advisory Role: Baxalta Advisory boards

Abstracts

Pelzer U.1, Wislocka L.1, Bischoff S.2, Jühling A.1, Sinn M.1, Striefler J.1, Klein F.3, Ghadjar P.4, Bahra M.3, Doerken B.1, Riess H.1, CONKO-SG Charité – Universitätsmedizin Berlin, Hämatologie/Onkologie/ Tumorimmunologie, Berlin, Germany, 2Charité – Universitätsmedizin Berlin, CONKO-SG, Berlin, Germany, 3Charité – Universitätsmedizin Berlin, Allgemein-, Visceral- und Transplantationschirurgie, Berlin, Germany, 4Charité – Universitätsmedizin Berlin, Radio-Onkologie und Strahlentherapie, Berlin, Germany 1

Introduction: The combination of nabpaclitaxel/gemcitabine (AG) is one standard of care for palliative therapy in patients (pts) with advanced pancreatic adenocarcinoma (APC). Only rare clinical data according feasibility and safety of AG in pts with elevated bilirubin levels are exist. This investigation focused on providing clinical data for those pts. Methods: Pts with APC treated with AG at our department between June 2012 and June 2015 were retrospectively screened according elevated total bilirubin (TB) levels. Other obligatory characteristics were the sufficient documentation of (non-) haematological toxicities, biliary tract stents, cancer resection, treatment sequence and overall survival (OS). Pts were primarily assigned to 3 groups according the TB level (I:TB ≥1.2-3xULN, II:TB>3-5xULN,III:TB:>5xULN). Results: 29/139 pts had elevated TB levels. Median age was 62[range 4278] years. 3 pts (10%) had primary curative resection, 13 (45%) had biliary tract stents. 26 pts (90%) had metastases at the time of diagnosis. Median treatment duration was 20 [1-442] days. Nab-paclitaxel was 1st/2nd line therapy for 10 pts (34%) respectively, 3rd line for 6pts (21%) and 4th line for 4pts (14%). 27 pts (93%) were treated in last line therapy. Median OS was 11.7 [95%CI: 7.9-14.0] months (m). Partitioned into the subgroups the mOS was for I: 11.8 [95%CI: 6.5-16.7] m, II: 9.2 [95%CI: 1.1-NA], III: 11.8 [95%CI: 5.9-20.0], p = 0.825. Median OS from the first application of nab-paclitaxel in these high-risk pts was for I: 2.6 [95%CI: 1.3-6.2], II: 1.1 [95%CI: 0.9-NA], III: 5.8[95%CI: 1.5-7.8], p = 0.129. Conclusions: The treatment with AG in pts with elevated serum bilirubin levels was feasible and safe in our cohort, if dosages were adapted on an individual manner regulated by caution of occurring toxicities. If these conditions are considered, the potential benefit of the treatment with AG also can be offered pts with hyperbilirubinemia. Disclosure: No conflict of interest disclosed. P867

Detection of cell-free mutated tumor DNA in patients with pancreatic cancer: a single center experience Hussung S.1, Hipp J.2, Köhler K.1, Michalczyk S.1, Fritsch K.1, Philipp U.1, von Bubnoff N.1, Follo M.1, Wittel U.2, Fritsch R.1 Uniklinik Freiburg/Innere Medizin/Hämatologie, Onkologie und Stammzelltransplantation, Freiburg, Germany, 2Uniklinik Freiburg, Abteilung für Allgemein- und Viszeralchirurgie, Freiburg, Germany 1

Introduction: Despite some recent progress through the introduction of more efficacious combination chemotherapy regimens, the clinical management of patients with pancreatic ductal adenocarcinoma (PDAC) has remained extremely challenging. Relapse rates following surgery for limited stage disease remain unacceptably high, and no predictive biomarkers exist to guide the optimal choice of systemic therapy for patients with advanced disease. The analysis of cell-free mutated tumor DNA (ctDNA) from plasma samples has been hailed as a breakthrough for the clinical management of difficult-to-treat tumor entities including PDAC. We therefore aim to explore the clinical utility of ctDNA measurement in PDAC patients presenting with limited or advanced stage disease at our university hospital center. Methods: We developed highly sensitive mutation-specific ddPCR assays for the 9 most common KRAS mutations in PDAC. We optimized extraction of cell-free DNA through adaption of commercially available systems.

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We obtain informed consent and plasma samples from patients during routine visits. Total cell-free DNA content, fragment length distribution, and the abundance of mutated vs. wild type KRAS are analyzed for each sample. Clinical and radiological parameters are analyzed in parallel. Results: We here report first preliminary data from an ongoing investigation. Out of 14 patients with resectable PDAC, eight patients (57%) were positive for circulating KRAS mutations. 5/6 patients with metastatic disease were highly positive for plasma KRAS mutations prior to treatment. 7 out of 12 patients undergoing treatment were or became positive during disease course. An increase of plasma KRAS mutations preceded clinical and radiological disease progression in several cases. ctDNA positivity also occurred in CA19-9 negative patients. Conclusion: Analysis of circulating KRAS mutations in the plasma of pancreatic cancer patients by ddPCR is a novel and inexpensive molecular monitoring tool. Integration into clinical routine is straightforward even without prior KRAS tumor sequencing. Circulating KRAS mutations can be readily detected in a high proportion of patients across all stages of disease. Early data indicate good correlation with clinical parameters and utility as monitoring tool for minimal residual disease. Prospective clinical trials will be required to establish whether and how ctDNA measurement can guide clinical decision-making and improve outcome for patients. Disclosure: No conflict of interest disclosed. P868

Treatment and survival of patients with advanced pancreatic cancer in community-based oncology group practices 2012 2015 Weide R.1, Burkhard O.2, Reiser M.3, Ehscheidt P.4, Feiten S.5 Praxisklinik für Hämatologie und Onkologie, Koblenz, Germany, 2Internistische Gemeinschaftspraxis Hämatologie, Onkologie, Palliativmedizin, Worms, Germany, 3pioh – Praxis Internistische Onkologie und Hämatologie, Frechen, Germany, 4Praxis für Hämatologie und Onkologie, Neuwied, Germany, 5Institut für Versorgungsforschung in der Onkologie, Koblenz, Germany 1

Introduction: Evaluation of treatment strategies and outcome of patients with advanced pancreatic cancer who received their treatment in oncology group practices in Germany since the introduction of FOLFIRINOX and gemcitabine+nab-paclitaxel. Comparison of overall survival between different treatment periods (1995 - 2004 versus 2012 - 2015). Methods: All consecutive patients with advanced pancreatic cancer who were treated between 01/2012 - 12/2015 in 4 oncology group practices were analysed retrospectively concerning treatment and outcome. Data were collected from patient files into a database and analysed statistically using SPSS. Results: 334 patients were analysed. Median age at diagnosis was 70 (32 - 94). 44% were female, 56% were male. 94 (28%) had locally advanced disease, 240 (72%) had distant metastases. 273 patients (82%) received some form of cytoreductive therapy, 61 (18%) best supportive care only. First line therapy was gemcitabine mono in 88 patients (32%), gemcitabine+nab-paclitaxel in 69 (25%), gemcitabine+erlotinib in 52 (19%) and FOLFIRINOX in 40 (15%). Radiotherapy was applied in 1 (0.4%). 105 patients (31%) received second line therapy consisting of gemcitabine+nab-paclitaxel in 23%, FOLFOX in 22%, gemcitabine mono in 19% and FOLFIRINOX in 9%. Third line therapy was given to 45 patients (13%) consisting of gemcitabine+nab-paclitaxel in 29%, FOLFOX in 18%, FOLFIRI in 13% and gemcitabine mono in 9%. 13 patients (4%) received some form of fourth line therapy. The median overall survival of the whole cohort is 36 weeks (1.7 - 365.6+). Median overall survival of patients receiving cytoreductive therapy is 37 weeks (3 - 209.3) compared to 25 weeks (1.7 - 365.6+) for patients receiving best supportive care only. Median overall survival for patients with locally advanced disease is 59.3 weeks (4.1 - 365.6+) compared to 30.4 weeks (1.7 - 166.6) for patients with distant metastases. Compared to our previous report 1995 - 2004 (J Support Oncol, 2004:159-163) the median overall survival for patients receiving cytotoxic treatment was reduced from 42 weeks to 37 weeks (3 209.3) for patients treated between 2012 - 2015.

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Conclusion: 82% of patients with advanced pancreatic cancer treated in community-based oncology group practices receive modern cytotoxic therapy, leading to an overall survival that compares favourably with prospective randomized trials. Survival seems to have not improved over time during the last 20 years in routine care. Disclosure: No conflict of interest disclosed. P869

Parenteral nutrition support for patients with advanced pancreatic cancer – interim results from the sub-project „PaNut“ of the Pancreatic Cancer Registry (TPK) Hegewisch-Becker S.1, Wolf T.2, Reiser M.3, Scheiner-Sparna R.4, Hamm D.4, Marschner N.5, for the TPK Registry Group Onkologische Schwerpunktpraxis Hamburg Eppendorf, Hamburg, Germany, BAG/Onkologische Gemeinschaftspraxis, Dresden, Germany, 3Praxis internistischer Onkologie und Hämatologie, Köln, Germany, 4iOMEDICO, Freiburg i. Br., Germany, 5Praxis für Interdisziplinäre Onkologie & Hämatologie, Freiburg i. Br., Germany 1 2

Introduction: In patients with pancreatic cancer weight loss and malnutrition are frequent and serious complications that may be associated with decreased quality of life (QoL) and a poorer prognosis. Malnutrition may be caused by loss of appetite and subsequent inadequate nutrient intake, exocrine pancreatic insufficiency, side effects of the anticancer therapy and/or the cancer-related anorexia-cachexia-syndrome induced by the tumor itself. Parenteral nutrition (PN) can be an option to counteract malnutrition Methods: The Tumor Registry Pancreatic Cancer (TPK) is a prospective, multicenter, observational study of pts with locally advanced or metastatic pancreatic cancer receiving systemic antineoplastic palliative therapy. Begin of pts recruitment was February 2014. Pts are recruited at the start of their palliative first-line treatment. A broad set of data regarding pts and tumor characteristics, comorbidities, factors affecting therapeutic decision making, previous and ongoing systemic treatments as well as outcome data are recorded. Within the sub-project “PaNut” additional data on PN are collected. Results: As of data cut-off 30.06.2015 451 pts were enrolled into the registry and 36 (8%) of these pts received additional PN, for 102 pts (23%) decision about PN was still not documented, 67% did not receive PN (2% data missing). Pts receiving PN had a lower BMI than the other pts with pancreatic cancer (mean BMI 22.2kg/m2 vs. 24.5kg/m2) and they were underweight for their age group. 53% of pts receiving PN were female. In 84% of pts PN started during first-line therapy. PN was initiated in median 3.6 weeks (25% quartile: 1.4 weeks, 75% quartile: 10.2 weeks) after start of therapy. Median duration was 13weeks (95% CI 4-17 weeks) and in most cases PN was continued until death. 18% of the pts received a mean energy supply of < 1,000kcal/day, 59% of 1,000-1,499kcal/day and 29% of >1,500kcal/day (multiple answers possible). About one third of the physicians stated that the application of a PN had lead to a better tolerability of the chemotherapy. One third of the physicians could not attest any influence of PN on chemotherapy tolerability. Conclusion: In routine clinical practice, at least 8% pts with locally advanced or metastatic pancreatic cancer receive PN. In pts receiving PN one third of the treating physicians stated a better tolerability of the chemotherapy. An update and further subgroup analyses will be presented. Disclosure: No conflict of interest disclosed.

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P870

Impact of immunohistochemistry for differentiation of primary liver cancer in clinical routine – Single center experience of 156 cases Hass H.1, Smith U.2, Schäffer M.3, Wellhäusser U.4, Denzlinger C.5, Scheurlen M.6, Markmann H.-U.7 Paracelsus-Klinik, Internistische Onkologie, Scheidegg, Germany, Onkologischer Schwerpunkt Stuttgart (OSP), Stuttgart, Germany, 3 Marienhospital Stuttgart, Viszeral- und Allgemeinchirurgie, Stuttgart, Germany, 4Marienhospital Stuttgart, Gastroenterologie, Stuttgart, Germany, 5 Marienhospital Stuttgart, Onkologie und Hämatologie, Stuttgart, Germany, 6 Universitätsklinik Würzburg, Gastroenterologie, Hepatologie und Onkologie, Würzburg, Germany, 7Marienhospital Stuttgart, Pathologie, Stuttgart, Germany 1 2

Background: Primary liver cancer (HCC / CCC) is characterized by a still unfavorable prognosis and reduced survival in most cases reasonable to a high recurrence rate. Despite of the different treatment options a fast and secure distinction between HCC and CCC is crucial for the affected patients. Reasonable to difficulties and pitfalls by diagnosis of primary liver cancer further analysis using immunohistochemistry (IHC) is needed in numerous cases. In this retrospective study the impact of IHC for discrimination of primary liver tumors were analyzed. Materials and methods: 156 primary liver tumors (120 HCC, 36 intrahepatic CCC) were analyzed by hematoxylin-eosin and immunohistochemical staining using molecular markers as HepPar-1, AFP, CD34, CK7, CK20, CA19-9 and CDX2 routinely. Significance of marker expression, sensitivity, specify and positive predictive value of the analyzed markers in relation to histological subtype were estimated using SPSS 10.0 Results: HepPar-1 was the most expressed marker in HCC and positive in 71.8% compared with 3% of cholangiocarcinoma (P < 0.0001). AFP staining was positive in 48.7% and again in the same patient (3%) with CCC (P < 0.0001). In this case histological examination showed a combined hepatocellular and cholangiocarcinoma with >50% of malignant cholangiocytes. The CD34 protein as a marker for vascular-associated tissue showed a positive reaction in 54.1% with HCC in comparison to 2 patients (6%) with cholangiocarcinoma (P < 0.0001). 86.9% with intrahepatic cholangiocarcinoma expressed CK7 and 50% expressed CK20. The results were significantly different from classical hepatocellular carcinoma (24.3% and 0%, respectively; P < 0.001 and P < 0.0001). Despite of one patient with combined HCC-CC (see above) AFP and HepPar-1 were in all cases negative. The carbohydrate antigen 19-9 (CA19-9) was positive in 55.5% of CCC and in 8% of hepatocellular carcinoma (P < 0.0001). Conclusions: Best predictive markers for differentiation between HCC and CCC were HepPar-1, CK7 and CA19-9. Using this panel a fast and secure differentiation by IHC was possible in over 95% in all analyzed cases. Disclosure: No conflict of interest disclosed. P871

Prognostic factors for tumor progress and recurrence of hepatocellular carcinoma (HCC) - A retrospective, single center study of 145 cases Hass H.1, Markmann H.-U.2, Schäffer M.3, Wellhäusser U.4, Scheurlen M.5, Smith U.6, Denzlinger C.7 Paracelsus-Klinik, Internistische Onkologie, Scheidegg, Germany, Marienhospital Stuttgart, Pathologie, Stuttgart, Germany, 3Marienhospital Stuttgart, Viszeral- und Allgemeinchirurgie, Stuttgart, Germany, 4Marienhospital Stuttgart, Gastroenterologie, Stuttgart, Germany, 5Universitätsklinik Würzburg, Gastroenterologie, Hepatologie und Onkologie, Würzburg, Germany, 6Onkologischer Schwerpunkt Stuttgart (OSP), Stuttgart, Germany, 7 Marienhospital Stuttgart, Onkologie und Hämatologie, Stuttgart, Germany 1 2

Materials and methods: Clinical data, tumor characteristics and the primary mode of management applied to our patients were analyzed using univariate and multivariate statistics. Results: During the period January 2004 till February 2014, 145 HCC patients presented to our multidisciplinary clinic (males 75%, mean age 68.8 ± 9.8 years). In 73.3% HCC developed on top of cirrhosis that was mainly due to chronic alcohol consumption (43.7%). In 32.2% of all patients an operative tumor resection in curative intention was performed. Median overall survival (OS) was 15 ± 19.2 months. Multivariate analysis identified tumor size (P = 0.001), grading (P = 0.002), proliferative activity (Ki67 level; P = 0.032), multifocal tumor (P = 0.045) and performed tumor resection (P < 0.0001) as independent prognostic factors for survival. Conclusions: Our data underline factors of tumor stage, tumor biology and performed surgical therapy in most cases as a consequence of liver function as independent prognostic factors for long-term OS in HCC. Disclosure: No conflict of interest disclosed. P872

Individualized therapy of advanced hepatocellular carcinoma by TACE and/or sorafenib: retrospective analysis using single center data Gläser D.1, Bartolomäus A.2, Lux C.2, Krammer-Steiner B.1 Klinikum Südstadt Rostock, Klinik für Innere Medizin III, Rostock, Germany, Klinikum Südstadt Rostock, Institut für Diagnostische Radiologie, Rostock, Germany 1 2

Introduction and objectives: Hepatocellular carcinoma (HCC) is globally the fifth most frequent malignancy and the third most frequent cause of malignancy-related death. Nevertheless, therapeutic options have been evoling over time. In addition to surgical and local treatment of early HCC stages, newer treatment options target advanced stage HCC. In addition, multimodal treatment strategies are being explored in clinicail trials. We aimed to retrospectively analyse patients from a single center who underwent treatment with systemic chemotherapy (sorafenib) or transarterial chemoembolization (TACE), or a combination thereof. Patients and methods: From data files covering 1/2009 to 12/2015, 56 patients with advanced HCC were identified who underwent treatment. TACE (doxorubicine/lipiodol) was performed in 16 patients, 26 patients underwent chemotherapy including sorafenib (2 x 400mg p.o. daily) and 14 patients underwent combined therapy consisting of TACE and chemotherapy. In these patients sorafenib was paused for one week before/after the intervention. Results: Treatment was administered until disease progression was evident or until worsening to ECOG < 2 in the supportive regimen. Median overall survival was 20.4 months with six patients continuing therapy at present. Adverse effects included weight loss exceeding 10 kg (21/56 patients), diarrhea (32/56 patients including three patients grade 3), mucositis (26/56 patients including two patients grade 3) and sorafenib-induced hand-foot syndrome (17/56 patients including one patient grade 3). Conclusions: Sequential treatment of advanced HCC employing local and systemic therapy is a feasible approach. Symptom control can be achieved at tolerable toxicity levels. Our retrospective analysis documented prolonged overall survival using the combined approach of TACE and chemotherapy compared to best supportiv care. Disclosure: No conflict of interest disclosed.

Background: Hepatocellular carcinoma (HCC) is a tumor with still unfavorable prognosis and reduced survival in most cases reasonable to a high recurrence rate. In this retrospective study prognostic factors were analyzed that affected long-term survival to improve future HCC management.

Abstracts

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P873

HER3-dependent resistance of gastric cancer cells against anti-proliferative effects of c-met inhibitors Jenke R.1,2, Rein M.1,2, Lordick F.2, Aigner A.1, Buech T.1 Universität Leipzig, Rudolf-Boehm-Institut für Pharmakologie und Toxikologie, Selbständige Abteilung für Klinische Pharmakologie, Leipzig, Germany, 2 University Cancer Center Leipzig (UCCL), Leipzig, Germany 1

Introduction: Inhibitors of the receptor tyrosine kinase (RTK) c-met are used as molecular-targeted therapeutics in gastric cancer. However, resistance may occur owing to the compensatory stimulation of other oncogenic RTKs. The aim of the present study was to delineate the role of RTKs of the HER family and their ligands in mediating resistance of gastric cancer cells against c-met inhibition. Methods: We used gastric cancer cell lines with a c-met amplification and evaluated the anti-proliferative effects of pharmacological or siRNA-mediated c-met inhibition as well as expression of HER family receptors and ligands after inhibitor treatment. Furthermore, we characterized the role of distal signaling molecules of the MAPK family by using phospho-antibody arrays and immunoblots. Results: RNAi-induced downregulation of c-met or the use of c-met kinase inhibitors led to a blockade of cellular proliferation and the disruption of tumor spheroid formation in anchorage-independent growth conditions. Without addition of c-met inhibitors, the investigated gastric cancer cells expressed HER1, HER2, and HER3 receptors and several HER1 ligands, but no HER3 ligands. After pretreatment with c-met inhibitors, no significant changes in expression of HER1 ligands or HER1 or HER2 receptors occurred. In contrast, HER3 expression was markedly upregulated after c-met inhibition. Of note, treatment of gastric cancer cells with the HER3 agonist heregulin restored the activity of p44/42 MAPK and Akt and partially reversed the anti-proliferative effects of c-met inhibitors. To further define the mechanism of HER3 upregulation upon c-met inhibition, we investigated the role of the transcriptional regulator protein SATB1 which in other tumor entities is known to be involved in the expression of HER receptors. Interestingly, downregulation of SATB1 abrogated the HER3 induction upon c-met inhibition and impaired the rescue effect of heregulin. Conclusions: Upon c-met inhibition, SATB1 is involved in the compensatory upregulation of HER3. Thus, targeting SATB1 may help to overcome resistance against c-met inhibition in gastric cancer. Disclosure: No conflict of interest disclosed. P874

Gene expression profiles of potential prognostic and predictive markers in disseminated gastric cancer Kipkeeva F.1, Muzaffarova T.1, Glubokova E.1, Narimanov M.2, Malehova O.2, Karpukhin A.1 Research Centre for Medical Genetics, Moscow, Russian Federation, 2NN Blokhin Russian Cancer Research Centre, Moscow, Russian Federation 1

Patients with gastric cancer have a poor prognosis since it is detected on the late stages in most cases. Therefore searching predictive and prognostic markers is important for understanding of tumor progression and developing personalized systematic therapy. In present study the selected clinical cases of disseminated gastric cancer was used. We investigated expression profiles of the genes :VEGF, VEGFR1, VEGFR2, NRP-1, bFGF, FGFR2, TGF-β, HER2/neu, TUBB, BRCA1, Ki67, PCNA by quantitative real-time PCR. Tumor and corresponding normal tissue was used in each case. Spirman coefficient was employed for data evaluation. An objective response to therapy with Herceptin was observed under increased expression of HER2 gene. Tubulin gene expression was associated with the effective action of taxanes (p = 0.036). The FGFR2 gene is under consideration as a new therapeutic target. However, the incidence of increased expression of FGFR2 was only 5%, which corresponds to the studied elsewhere its frequency amplification at gastric cancer.

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Most often elevated mRNA level in the tumor relative to the control was observed for NRP-1, VEGFR2 and TGF-β genes (32% - 41% ). The correlation of these gene expression levels was found for the first time (R = 0.63; p = 0.002). Survival more than 1 year was associated with simultaneously increased expression level of these genes (OR = 10.6; 95%CI =1.48 to 76.08; p = 0.018). An inverse correlation VEGF and bFGF gene expression was revealed also (R=-0.73; p = 0.005).This effect probably reflects alternative ways to stimulate the gastric cancer development and must be taken into account in the choice of means of targeted therapy for this disease. Disclosure: No conflict of interest disclosed. P875

Summary of biomarker data from 2 randomized, double-blind placebo-controlled phase 3 trials (RAINBOW and REGARD) of second-line Ramucirumab for advanced gastric or gastroesophageal junction carcinoma Lorenzen S.1, Thuß-Patience P.2, Hofheinz R.-D.3, Brück P.4, Emig M.4, Hozak R.R.5, Ferry D.6, Melemed S.7, Ouyang H.7, Fuchst C.S.8, Wilke H.-J.9 III. Medizinische Klinik, Klinikum rechts der Isar, München, Germany, Medizinische Klinik m.S. Hämatologie, Onkologie und Tumorimmunologie, Charité Berlin – Universitätsmedizin Berlin, Berlin, Germany, 3 TagesTherapieZentrum, Interdisziplinäres Tumorzentrum, Universitätsmedizin Mannheim, Mannheim, Germany, 4Medizinische Abteilung – Onkologie, Lilly Deutschland GmbH, Bad Homburg, Germany, 5Global Statistics, Eli Lilly and Company, Indianapolis, United States, 6Eli Lilly and Company, Oncology, Bridgewater, United States, 7Eli Lilly and Company, Oncology, Indianapolis, United States, 8Dana Farber Institute, Harvard Medical School, Department of Medical Oncology, Boston, United States, 9Klinikum Essen-Mitte, Medizinische Onkologie/Hämatologie, Essen, Germany 1 2

Introduction: Ramucirumab (RAM) is a human IgG-1 monoclonal antibody that targets the extracellular domain of vascular endothelial growth factor receptor 2 (VEGFR2). In 2 phase 3 trials, second-line RAM, given either as add-on to paclitaxel (RAINBOW, n = 665) or in monotherapy (REGARD, n = 355), significantly improved overall survival (OS) of patients (pts) with advanced gastric or gastroesophageal junction carcinoma compared to placebo (Wilke et al., Lancet Oncol 2014; Fuchs et al., Lancet 2014). This analysis summarizes the results of the angiogenic biomarker program from both trials in patients with evaluable plasma, serum or tissue samples. Methods: In RAINBOW, 29 biomarkers (incl. VEGF-C and -D, soluble (s) VEGFR1,2,3) were assessed in plasma from pts with evaluable samples (n = 257-380 per marker). In REGARD, 2 candidate tumor biomarkers (HER2 and VEGFR2) and 4 circulating biomarkers (VEGF-C and -D, sVEGFR1 and -R3) were assessed in pts with evaluable tissue (n = 152) or serum samples (n = 32). Due to small sample size, REGARD plasma markers were assessed by scatterplot. For other markers, pts were dichotomized into low and high expression subgroups (cutpoint: lower limit of quantification (LLOQ) for markers with >20% below LLOQ; median for all other). Additional exploratory cutpoints were assessed. Markers were correlated with clinical efficacy parameters and analyzed for predictive effects by interaction models, and for prognostic effects by main effect models. Results were correlated with patient clinical data (OS, progression-free survival [PFS]). Results: Neither RAINBOW nor REGARD identified strong predictive associations between the markers assessed and OS or PFS; REGARD serum results are limited by the small sample numbers. RAINBOW identified several pharmacodynamic trends: In the RAM arm, VEGF-D and placental growth factor levels increased and angiopoietin 2 levels decreased after start of RAM; these markers changed minimally in the placebo arm. Potential prognostic markers (low baseline levels associated with longer OS and PFS at alpha < 0.05) included: CRP, HGF, ICAM-3, IL-8, serum amyloid A, VCAM1. Conclusions: Although these exploratory biomarker analyses on tumor biopsies and plasma or serum samples available from RAINBOW and REGARD patients did not identify a significant predictive biomarker for RAM efficacy, several pharmacodynamic trends and potential markers for

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prognostic effects were observed, warranting further examination in the future. Disclosure: Sylvie Lorenzen: No conflict of interest disclosed. Hans-Jochen Wilke: Financing of Scientific Research: Eli Lilly and Company P876

Peritonealcarcinosis – Cytoreduktive surgery (CRC) and hyperthermic intraperitoneal chemotherapy (HIPEC) management and results in a specialized hospital Rudolph S.1, Meißner C.1, Borchert K.2, Kahl C.2, Ridwelski K.1 Klinikum Magdeburg gGmbH, Klinik für Allgemein- und Viszeralchirurgie, Magdeburg, Germany, 2Klinikum Magdeburg gGmbH, Klinik für Hämatologie und Onkologie, Magdeburg, Germany 1

Introduction: The peritoneal carcinosis is a sign of an advanced tumor disease with poor prognose. For the treatment of peritoneal specific treatment has been developed since 1980th. This includes a combination of the most complete possible removal of the tumor (cytoreductive surgery – CRS) and a directly subsequent chemotherapy in the abdomen (HIPEC). In selected cases, surgical treatment in combination with HIPEC is possible. Methods: 1. First we need is a patient selection (general condition: ECOG < 2, no extraabdominal metastasis, max. 3 respectable liver metastases, not biliary obstruction, not small bowel involvement with multiple partial obstruction, peritoneal carcinosis in colon cancer, gastric cancer, mesothelioma, Pseudomyxoma the peritoneum, (ovarian cancer)). 2. diagnostic treatment (CT-Thorax-Abdomen, Laparoscopy to confirm the diagnosis and assessment of peritoneal cracinosis index). 3. Implementation procedure (exploration of the abdomen, determining the peritoneal index, possibly resection of the primary tumor, resektion oft he peritoneum and the omentum, electrosurgical destruction not resectable tumor nodules in the peritoneum visceral, resection of affected organs (colon, spleen, pancreas tail, stomach, hepatectomy), closed HIPEC about 4 - 5 drainages at 40-42 °C) Results: 11/2011 to 04/2016, we performed 22 interventions. 59 years (3776) was the average age of the patients. ECOG 0-1. (Colon 12x, Stomach 3x, Psydomyxoma 3x, Ovar-Tube-peritoneum 3x, Mesotheliom 1x). The 30 Days-legality of the colon patients n = 1. The group of colon cancers have a current survival rate of 21.7 months (still alive). Summary: CRC and HIPEC is a therapeutic option in which also longterm survival is possible in selected patients. Decisive for the results is the selection of patients and the experience in operative and perioperative management. Disclosure: No conflict of interest disclosed.

Methods: With the aim to receive additional information on mechanism of clear cell renal cell carcinoma (ccRCC) progression and development biomarkers we studied two groups of genes. The first was made up of genes with the most expression. By bioinformatic analysis of gene expression databases 200 genes were selected. After quantitative RT-PCR of ccRCC sample in comparison with normal renal tissue 21 genes with the most frequent increased expression were identified. The majority of these genes were direct targets of HIF1α. Results: It was found an association of six gene decreased expression (CA9, NDUFA4L2, VWF, IGFBP3, EGLN3, BHLHE41) with overall survival (OS) and genes VWF, IGFBP3,STC2 with metastasis (p = 0,008 0,044). Among these genes were as novel, without established significance in ccRCC, as NDUFA4L2 (OR = 0,048; 95% CI: 0,005 - 0,444), so well known biomarkers, as CA9. The second group was formed of genes participate in angiogenesis (5 genes), PI3K-AKT-mTOR pathway (10 genes) and EGFR. Only VEGFR1 and VEGFA decreased expression were interconnected with both OS and metastasis (p = 0,009 - 0,023). Increased expression in significant association with OS was observed for SAA1 and three genes – CSF1R, Fn1 and C1QA tended to be increased. All these genes are associated with inflammatory response. Conclusions: The data give a possibility for proposition of consistent involvement HIF1A -> HIF2A -> NF-kB/STATs modules, which can lead to the development of metastasis and reduced OS. Thus, for the first time shown coordinated expression decreasing of number of regulated HIF1α genes in connection with OS and metastasis. Disclosure: No conflict of interest disclosed. P878

Correlation of response with overall survival (OS) for nivolumab vs everolimus in advanced renal cell carcinoma (aRCC): Results from the phase III CheckMate 025 study Grünwald V.1, Motzer R.J.2, Sharma P.3, Escudier B.4, McDermott D.F.5, George S.6, Srinivas S.7, Tykodi S.S.8, Sosman J.A.9, Plimack E.R.10, Nathan P.11, Tomita Y.12, Zhao H.13, Waxman I.M.14, Hammers H.J.15 Hannover Medical School, Hannover, Germany, 2Memorial Sloan Kettering Cancer Center, New York, United States, 3MD Anderson Cancer Center, University of Texas, Houston, United States, 4Institut Gustave Roussy, Villejuif, France, 5Beth Israel Deaconess Medical Center, Dana-Farber/Harvard Cancer Center, Boston, United States, 6Roswell Park Cancer Institute, Buffalo, United States, 7Stanford Cancer Institute, Stanford, United States, 8University of Washington and Fred Hutchinson Cancer Research Center, Seattle, United States, 9Vanderbilt University Medical Center, Nashville, United States, 10Fox Chase Cancer Center, Philadelphia, United States, 11Mount Vernon Cancer Centre, Middlesex, United Kingdom, 12Niigata University, Niigata, Japan, 13 Bristol-Myers Squibb, Pennington, United States, 14Bristol-Myers Squibb, Lawrenceville, United States, 15Johns Hopkins Sidney Kimmel Comprehensive Cancer Center, Baltimore, United States 1

Introduction: Gene expression profiling is a promising technique for refining the diagnosis and staging of RCC, as well as for highlighting potential therapeutic targets.

Introduction: In CheckMate 025 (NCT01668784), nivolumab demonstrated superior OS (25.0 mo; 95% CI, 21.8-not estimable [NE]) vs everolimus (19.6 mo; 95% CI, 17.6-23.1) and a higher objective response rate (25% vs 5%, P < 0.001) in previously treated patients with aRCC (N Engl J Med 2015;373:1803-13). Here, we investigated OS benefit with nivolumab vs everolimus by tumor response. Methods: Patients were randomized 1:1 to nivolumab 3 mg/kg intravenously every 2 weeks or everolimus 10 mg orally once daily. We conducted a landmark analysis of OS by best response (complete/partial response [responders], stable disease [SD], and progressive disease [PD]) within 4 months of randomization. OS was assessed using the Kaplan-Meier method. Results: For the 410 and 411 patients randomized to nivolumab and everolimus, median time to response was 3.5 months (range: 1.4-24.8) and 3.7 months (1.5-11.2), respectively. By month 4 with nivolumab, 19% of patients were responders, 30% had SD, and 40% had PD; with everolimus, 3% of patients were responders, 45% had SD, and 31% had PD. The remainder of patients had discontinued, died, or had an unevaluable response by 4 months. Median (95% CI) OS is shown (table). For nivolum-

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–294

Posterdiskussion

Nierentumoren, Prostatakarzinom P877

The features of quantitative expression profiles of gene cluster in renal cell cancer Apanovich N.V.1, Poyrkov S.V.1, Peters M.V.2, Korotaeva A.A.1, Markova A.S.2, Kamolov B.S.2, Pronina I.V.1, Braga E.A.1, Matveev V.B.2, Karpukhin A.V.1 Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Moscow, Russian Federation, 2Cancer Research Centre, Russian Academy of Medical Sciences, Moscow, Russian Federation 1

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ab, OS rates at 12 and 18 months were 96% and 89% in responders, 91% and 81% in patients with SD, and 70% and 50% in patients with PD by 4 months. For everolimus, OS rates at 12 and 18 months were 93% and 77% in responders, 90% and 79% in patients with SD, and 54% and 35% in patients with PD by 4 months. Analyses of each subgroup by disease features will be presented. Conclusions: Patients who had a best response of SD within 4 months of starting treatment had a similar survival trajectory compared with responders. Patients with a best response of progression had an improved OS with nivolumab vs everolimus. Tab. 1.

Nivolumab, n (%) Median OS (95% CI) 12-month rate, % (95% CI) 18-month rate, % (95% CI) Everolimus, n (%) Median OS (95% CI) 12-month rate, % (95% CI) 18-month rate, % (95% CI)

Responders SD 77 (19) 121 (30) NE (24.1-NE) NE (22.7-NE)

PD 164 (40) 14.0 (11.3-16.9)

96 (88–99)

91 (84–95)

70 (63–77)

89 (80–95)

81 (73–87)

50 (42–57)

Results: Baseline QoL was collected for 86% of all randomized patients; 361/410 (nivolumab) and 343/411 (everolimus). Patients with FKSI-DRS improvement and time to improvement are shown (table). A positive correlation was observed between FKSI-DRS baseline scores and OS (correlation coefficient, 0.27; P ≤ 0.001) in the overall population (n = 705). A high baseline QoL (FKSI-DRS score above median) and 2-point QoL improvement from baseline was associated with improved OS (median OS not estimable [NE] [95% CI, 28.1-NE]) up to 12 weeks. Results using the EQ-5D questionnaire of general health status will be presented. Conclusions: In this global study of patients with aRCC, treatment with nivolumab vs everolimus resulted in a significantly greater proportion of patients experiencing meaningful QoL improvement. These results suggest that assessment of QoL may be helpful to evaluate the extent of OS benefit in clinical practice. Tab. 1.

14 (3) 186 (45) 127 (31) NE (12.4-NE) 25.0 (22.9-NE) 9.8 (6.1-12.2)

93 (59–99)

90 (85–93)

54 (45–63)

77 (45–92)

79 (72–84)

35 (27–43)

FKSI-DRS

Disclosure: Viktor Grünwald: Advisory Role: Bristol-Myers Squibb, Pfizer, Novartis; Financing of Scientific Research: Bristol-Myers Squibb, Pfizer, Novartis; Other Financial Relationships: Bayer, Bristol-Myers Squibb, Pfizer, Novartis Hans Hammers: Advisory Role: Bristol-Myers Squibb, Exelixis, Pfizer, Cerulean; Expert Testimony: SFJ, Bristol-Myers Squibb, Exelixis, Newlink, Pfizer, GSK, Tracon

Nivolumab

Everolimus

Patients Time to with improveimprovement ment mo n/N (%) (95% CI) 200/361 4.7 (55.4) (3.7–7.5)

Patients Time to with improveimprovement ment mo n/N (%) (95% CI) 126/343 NE (36.7) (NE-NE)

Nivolumab vs Everolimus HR (95% CI)

P value 1.66 (1.33– 2.08) P < 0.001

Disclosure: Viktor Grünwald: Advisory Role: Bristol-Myers Squibb, Pfizer, Novartis; Financing of Scientific Research: Bristol-Myers Squibb, Pfizer, Novartis; Other Financial Relationships: Bayer, Bristol-Myers Squibb, Pfizer, Novartis Robert Motzer: Advisory Role: Novartis, Eisai, Pfizer; Expert Testimony: Exelixis, Bristol-Myers Squibb, Novartis, GSK, Eisai, Pfizer P880

P879

Improvements in quality of life and overall survival in patients with advanced renal cell carcinoma treated with nivolumab vs everolimus in the phase III CheckMate 025 study Grünwald V.1, Cella D.2, Nathan P.3, Doan J.4, Dastani H.5, Taylor F.6, Bennett B.7, DeRosa M.6, Berry S.8, Broglio K.8, Berghorn E.9, Motzer R.J.10 Hannover Medical School, Hannover, Germany, 2Northwestern University, Chicago, United States, 3Mount Vernon Cancer Centre, Middlesex, United Kingdom, 4Bristol-Myers Squibb, Wallingford, United States, 5Bristol-Myers Squibb, Lawrenceville, United States, 6Adelphi Values, Boston, United States, 7 Adelphi Values, Bollington, United Kingdom, 8Berry Consultants, Austin, United States, 9Bristol-Myers Squibb, Wallington, United States, 10Memorial Sloan Kettering Cancer Center, New York, United States 1

Introduction: In the CheckMate 025 study (NCT01668784), previously treated patients with advanced renal cell carcinoma (aRCC) randomized to nivolumab had an overall survival (OS) benefit vs everolimus (hazard ratio [HR], 0.73; 98.5% CI, 0.57-0.93). Median scores on the Functional Assessment of Cancer Therapy-Kidney Symptom Index-Disease Related Symptoms (FKSI-DRS) scale increased from baseline and were significantly (P < 0.05) improved with nivolumab vs everolimus through week 104 (N Engl J Med 2015;373:1803-13). Here we compare quality of life (QoL) between treatment arms and explore the relationship with OS. Methods: QoL was measured by the kidney-specific FKSI-DRS questionnaire at baseline and every 4 weeks. Descriptive statistics and multivariate analyses were used to analyze between-arm differences. Chi-square analyses were used to assess differences in the proportion of patients who experienced meaningful improvement (2-point change) with nivolumab vs everolimus. Association between baseline QoL and OS was assessed using the Pearson correlation coefficient method, and change from baseline QoL and OS using a previously described method (J Clin Oncol 2003;21:1536-43).

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Evaluation of safety, tolerability and activity of Axitinib in patients (pts) with advanced or metastatic renal cell carcinoma (a/mRCC) in routine clinical practice: the STAR-TOR registry Bergmann L.1, Woike M.2, Krekeler G.2, Steiner T.3, Goebell P.J.4, Göhler T.5, Harich H.-D.6, Herrmann E.7, Rebmann U.8, Kalanovic D.2 J.-W.-Goethe-Universität, Medizinische Klinik II, Frankfurt, Germany, 2Pfizer Pharma GmbH, Berlin, Germany, 3Helios-Klinikum, Urologie, Erfurt, Germany, 4 AURONTE (Urologische Klinik und Hämatologie/Onkologie (Med. 5)), Universitätsklinikum Erlangen, Erlangen, Germany, 5Onkozentrum, Dresden, Germany, 6Onkologie Hof – Medizinisches Versorgungszentrum GmbH, Hof, Germany, 7Universitätsklinikum Münster, Urologie, Münster, Germany, 8 Diakonissenkrankenhaus Dessau, Urologie, Dessau, Germany 1

Introduction: Axitinib (AXI), an oral tyrosine kinase inhibitor, is approved in the EU for the treatment of pts with a/mRCC after failure of prior therapy with sunitinib or a cytokine. A pivotal study had demonstrated significantly increased progression-free survival (PFS) with AXI compared to Sorafenib (6.7 vs. 4.7 months (mo); one-sided p < 0.0001, log-rank). Here we present first preliminary data from a non-interventional post-approval study. Methods: A German multicenter registry for pts with a/mRCC (NCT00700258) was amended in October 2012 with regulatory and ethic committee´s approval to include pts treated with AXI. Objectives are the evaluation of the safety profile, the tolerability and anti-tumor activity of AXI as well as the profile, comorbidity and characteristics of pts and the sequence of systemic therapies in pts with a/mRCC. Inclusion criteria are histologically confirmed a/mRCC treated with AXI and written informed consent. Results: From November 2012 to February 2016, 64 study sites recruited 154 AXI pts. Characteristics: 70.4% male, median age 67.5 years (28.084.0), median Karnofsky index 80% (50-100%). Histological subtype: 77.3% clear cell only, 22.7% other histologies. Median duration of disease

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until onset of AXI treatment was 3.0 years. In 48.7% of pts AXI was used as second-line therapy, in 26.6% as third-line treatment and in 22.0% as fourth line and beyond. 4 pts were treated in first line. Median number of prior therapies was 1. 85 pretreated pts were evaluable with regard to MSKCC criteria: 10.6% favorable, 34.1% intermediate and 55.3% poor risk. For all 154 pts, adverse and serious adverse events were observed in 73.4% and 38.3% of pts (drug related in 55.2% and 14.9% of pts), respectively. Drug-related toxicities (incidence ≥ 10%) of any grade were observed in the following categories: gastrointestinal disorders (36.4%), general disorders (23.4%), skin and subcutaneous tissue disorders (12.3%), metabolic and nutritional disorders (11.0%). Median PFS of the total study cohort was 6.4 mo, median PFS for the subgroup of second line pts (n = 75) was 5.7 mo. Overall survival for all 154 pts was 18.3 mo. Conclusions: These preliminary data show that AXI seems to be a an effective therapy option for a/mRCC pts in second and higher therapy lines in the routine clinical setting. Toxicities were generally manageable, their incidence being lower than in published phase III data. PFS and OS were in the expected range. Disclosure: Lothar Bergmann: Advisory Role: Pfizer, Novartis, Bristol Myers Squibb; Expert Testimony: Pfizer, Novartis, Roche, Exelixis Daniel Kalanovic: Employment or Leadership Position: Pfizer Pharma GmbH P881

Evaluation of safety, tolerability and activity of Sunitinib in patients (pts) with advanced or metastatic renal cell carcinoma (a/mRCC) in routine clinical practice: the STAR-TOR registry Bergmann L.1, Woike M.2, Krekeler G.2, Steiner T.3, Goebell P.J.4, Göhler T.5, Harich H.-D.6, Herrmann E.7, Rebmann U.8, Kalanovic D.2 J.-W.-Goethe-Universität, Medizinische Klinik II, Frankfurt, Germany, 2Pfizer Pharma GmbH, Berlin, Germany, 3Helios-Klinikum, Urologie, Erfurt, Germany, 4 AURONTE (Urologische Klinik und Hämatologie/Onkologie (Med. 5)), Universitätsklinikum Erlangen, Erlangen, Germany, 5Onkozentrum, Dresden, Germany, 6Onkologie Hof – Medizinisches Versorgungszentrum GmbH, Hof, Germany, 7Universitätsklinikum Münster, Urologie, Münster, Germany, 8 Diakonissenkrankenhaus Dessau, Urologie, Dessau, Germany 1

Introduction: Sunitinib (SUN), an oral tyrosine kinase inhibitor, is approved in the EU for the treatment of pts with a/mRCC. A pivotal study had demonstrated significantly increased progression-free survival (PFS) with SUN compared to the former standard interferon alpha (11 vs. 5 months); p < 0.001). To evaluate the safety profile and efficacy of SUN in a clinical routine setting, data collection in a post-approval non-interventional trial seems to be appropriate. Methods: A German multicenter registry for pts with a/mRCC (NCT00700258) was amended to include SUN pts in June 2010 with regulatory and ethic committee´s approval. Objectives are the evaluation of the safety profile, the tolerability and anti-tumor activity of SUN as well as the profile, comorbidity and characteristics of pts and the sequence of systemic therapies in pts with a/mRCC. Inclusion criteria are histologically confirmed a/mRCC treated with SUN and written informed consent. Results: From July 2010 to February 2016, 105 active study sites recruited 436 pts for this interim analysis. Characteristics: 72.8% male, median age 67.8 years (31.0-87.7), median Karnofsky index 90% (40-100%). Histological subtype: 81.9% clear cell only, 18.1% other histologies. In 78.4% of pts SUN is used as first-line therapy. 280 pts were evaluable with regard to MSKCC criteria: 11.8% favorable, 62.9% intermediate, 25.4% poor risk. For all 436 pts, adverse and serious adverse events were observed in 76.1% and 35.6% of pts (drug related in 60.3% and 14.0% of pts), respectively. Most common drug-related toxicities (incidence ≥ 10%) of any grade were gastrointestinal disorders (36.0%), general disorders (24.8%), skin and subcutaneous tissue disorders (19.5%), blood and lymphatic system disorders (17.4%), and nervous system disorders (14.9%). Median PFS for clear-cell a/mRCC pts (n = 363) was 8.6 months (mo), for pts with other histologies 4.2 mo. The subgroup of clear-cell a/mRCC pts for which SUN was first-line therapy (n = 285) had a PFS of 9.2 mo, for second line clear-

Abstracts

cell pts (n = 49) 7.0 mo, ≥ third line (n = 28) 5.6 mo. Overall survival (OS) for the entire study cohort was 22.1 mo. Conclusions: The patient population in the registry represents the expected pattern in pts with a/mRCC regarding distribution of age, sex, and histology. Efficacy and tolerability of SUN in routine clinical practice confirms published data Disclosure: Lothar Bergmann: Advisory Role: Pfizer, Novartis, Bristol Myers Squibb; Expert Testimony: Pfizer, Novartis, Roche, Exelixis Daniel Kalanovic: Employment or Leadership Position: Pfizer Phama GmbH P882

Evaluation of safety, tolerability and activity of Temsirolimus in patients (pts) with advanced or metastatic renal cell carcinoma (a/mRCC) in routine clinical practice: the STAR-TOR registry Bergmann L.1, Woike M.2, Krekeler G.2, Steiner T.3, Goebell P.J.4, Göhler T.5, Harich H.-D.6, Herrmann E.7, Rebmann U.8, Kalanovic D.2 J.-W.-Goethe-Universität, Medizinische Klinik II, Frankfurt, Germany, 2Pfizer Pharma GmbH, Berlin, Germany, 3Helios-Klinikum, Urologie, Erfurt, Germany, 4 AURONTE (Urologische Klinik und Hämatologie/Onkologie (Med. 5)), Universitätsklinikum Erlangen, Erlangen, Germany, 5Onkozentrum, Dresden, Germany, 6Onkologie Hof – Medizinisches Versorgungszentrum GmbH, Hof, Germany, 7Universitätsklinikum Münster, Urologie, Münster, Germany, 8 Diakonissenkrankenhaus Dessau, Urologie, Dessau, Germany 1

Introduction: Temsirolimus (TEM), an i.v. mTOR inhibitor, is approved in the EU for the first-line treatment of pts with a/mRCC who have at least 3 of 6 prognostic risk factors. A pivotal study had demonstrated significantly increased overall survival (OS) with TEM in poor risk pts compared to the former standard interferon alpha (10.9 vs. 7.3 months (mo); p = 0.008). To evaluate the safety profile and efficacy of TEM in a clinical routine setting, collection of data in a post-approval non-interventional trial is useful. Methods: A German multicenter registry for pts with a/mRCC treated with TEM was started in January 2008 (NCT00700258) with regulatory and ethic committee´s approval. Objectives are the evaluation of the safety profile, the tolerability and anti-tumor activity of TEM as well as the profile, comorbidity and characteristics of pts and the sequence of systemic therapies in pts with a/mRCC. Inclusion criteria are histologically confirmed a/mRCC treated with TEM and written informed consent. Results: From February 2008 to February 2016, 118 active study sites recruited 568 pts. Characteristics: 68.1% male, median age 68.7 years (34.987.0), median Karnofsky index 80% (40-100%). Histological subtype: 74.3% clear cell, 11.4% papillary, and 1.9% chromophobe RCC. 196 pts were evaluable with regard to modified MSKCC criteria. 78.6% of these pts are classified as poor risk. Median number of metastatic sites is 2 (0-6). Adverse and serious adverse events are observed in 74.8% and 43.8% of pts (drug related in 42.1% and 9.2% of pts), respectively. Drug-related toxicities (incidence ≥ 10%) of any grade were observed in the following categories: general disorders (17.4%), skin and subcutaneous tissue disorders (15.7%), gastrointestinal disorders (13.6%), blood and lymphatic system disorders (11.4%). Median progression-free survival for the total patient population is 4.2 mo, for the subgroup of first line poor risk pts (n = 154) 3.2 mo. A subgroup analysis of pts with baseline LDH ≤ 300 U/l (n = 316) vs > 300 U/l (n = 115) shows significant longer median OS for the low LDH group (12.1 vs. 7.2 mo, p = 0.007). Median OS for all pts is 11.1 mo. Conclusions: The patient population in the registry represents the expected pattern in pts with a/mRCC regarding distribution of age, sex, and histology. Safety and efficacy of TEM in routine clinical practice confirm current phase III data. Baseline LDH > 300 U/l seems to be a negative prognostic parameter for OS. Disclosure: Lothar Bergmann: Advisory Role: Pfizer, Novartis, Bristol Myers Squibb; Expert Testimony: Pfizer, Novartis, Roche, Exelixis Daniel Kalanovic: Employment or Leadership Position: Pfizer Pharma GmbH

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The PAZOREAL study: A non-interventional study to asses efficacy and safety of Pazopanib and everolimus in a real life setting: reflecting a changing mRCC treatment landscape Goebell P.J.1, Doehn C.2, Grüllich C.3, Grünwald V.4, Steiner T.5, Welslau M.6 University Erlangen, Department of Urology, Erlangen, Germany, 2Urologikum Lübeck, Lübeck, Germany, 3National Center for Tumor Diseases (NCT), Department of Medical Oncology, Heidelberg, Germany, 4Medical School of Hannover, Department of Hematology, Hemostaseology, Oncology and Stem Cell Transplantation, Hannover, Germany, 5HELIOS Klinikum Erfurt, Department of Urology, Erfurt, Germany, 6Medical Center – Aschaffenburg, Aschaffenburg, Germany 1

Introduction: Renal cell carcinoma (RCC) is diagnosed in about 15,500 patients per year in Germany, with up to 60% of patients requiring systemic treatment in the metastatic setting. VEGF inhibition as well as mTOR inhibition are promising therapeutic methods and are broadly used in metastatic renal cell carcinoma (mRCC) therapy. Pazopanib 1st line, followed by Everolimus 2nd line is a well-established standard. Recently, Nivolumab has been approved in Europe for use after previous therapy. This non-interventional study is designed to closely observe real-life use of the evolving therapy environment in 1st, 2nd and 3rd line in patients starting with Pazopanib in 1st line. Methods: PAZOREAL is a prospective, multi-center, non-interventional study to evaluate efficacy, tolerability, safety, and quality of life in patients with mRCC treated with Pazopanib in 1st line, Nivolumab or Everolimus in 2nd line, or Everolimus in 3rd line after Nivolumab. Adults with a histologically confirmed diagnosis of mRCC of any subtype , who have a life expectancy of at least 6 months, and whose systemic treatment is decided to start on 1st line Pazopanib or to continue on 3rd line Everolimus after 2nd line Nivolumab, are eligible. Treatment has to be planned according to the respective German drug labels. Patients’ history, treatment, disease assessment, concomitant medication, adverse events, follow-up treatments and survival are documented. Variables include the time on drug (TD) for documented therapies, overall survival (OS), dosing parameters, safety and QoL (assessed by EQ-5D-5L questionnaire). Recruitment of patients has started in December 2015 and is planned to end in December 2018, with a goal of 450 documented patients at about 150 sites in Germany. Four interim analyses are planned; the first will be performed in April 2017. Immediately after approval of Nivolumab in Germany, the study was opened for documentation of therapy sequences including in-label Nivolumab treatment in 2nd line. Disclosure: Peter Goebell: Advisory Role: Teilnahme an Advisory-boards: Bayer, Bristol-Myers-Squibb, Novartis, Pfizer, Janssen, Sanofi-Aventis; Financing of Scientific Research: Vortragshonorare: Astellas, Bayer, Bristol-Myers-Squibb, Novartis, Pfizer, Janssen, Sanofi-Aventis; Other Financial Relationships: Reisekostenunterstützungen: Astellas, Bayer, Bristol-Myers-Squibb, Novartis, Pfizer, Janssen, Sanofi-Aventis Manfred Welslau: Advisory Role: Novartis, GSK, Pfizer, BMS P884

EVI1 - a novel oncogene in prostate carcinoma Wang H.1,2, Queisser A.3, Hagedorn S.3, Konantz M.1, Schäfer T.1, Alavi S.3, Vogel W.4,5, Mässenhausen A.V.3, Kristiansen G.3, Duensing S.6, Kirfel J.3, Perner S.4,5, Lengerke C.1,2,7 University Hospital Basel, Department of Biomedicine, Basel, Switzerland, University of Tuebingen Medical Center II, Department of Hematology & Oncology, Tuebingen, Germany, 3University Hospital of Bonn, Section for Prostate Cancer Research, Institute of Pathology, Center for Integrated Oncology Cologne/Bonn, Bonn, Germany, 4Pathology of the University Hospital of Luebeck, Luebeck, Germany, 5The Leibniz Research Center Borstel, Borstel, Germany, 6University of Heidelberg School of Medicine, Section of Molecular Urooncology, Department of Urology, Heidelberg, Germany, 7University Hospital Basel, Clinic for Hematology, Basel, Switzerland 1 2

Introduction: Prostate cancer (PCa) is the most commonly diagnosed non-cutaneous cancer in men in the western world. Ecotropic Viral Integration site 1 (EVI1) is a transcriptional regulator mainly known as a

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hematopoietic stem cell marker and for its role in acute myeloid leukemia aggressiveness. However, EVI1 expression has been also documented in various solid tumors. Here we investigate expression and roles of EVI1 in prostate carcinogenesis. Methods: The expression profile of EVI1 was investigated in benign prostatic glands and a PCa progression cohort containing 148 primary PCa samples, 39 lymph node metastases and 32 castration resistant prostate carcinomas (CRPCs) by immunohistochemistry (IHC). Cells from the human prostate carcinoma cell line PC3 were stably transduced with lentiviral plasmids containing shRNAs against EVI1. Knockdown and corresponding control cells were then analyzed in cell proliferation (xCELLigence system, Ki67 staining), cell cycle, EdU, migration, tumor sphere, apoptosis (PI and Annexin-V/7-AAD staining), and in vivo zebrafish xenotransplant tumorigenicity assays and analyzed by qRT-PCR and immunoblot. Docetaxel resistant PC3 cells were generated by long-term in vitro treatment (6 months). Results: In healthy prostatic tissue, EVI1 expression was confined to the prostate stem cell compartment located at the basal layer and identified by the stem cell marker CD44. In the PCa progression cohort, heterogeneous EVI1 expression was found but EVI1 staining strongly increased with tumor progression (p < 0.001), suggesting EVI1 activation as a driver event. Functionally, EVI1 knockdown inhibited proliferation and cell cycle progression of PC3 cells. EVI1 knockdown cells accumulated in the G0/G1 phase and showed suppression of pMAPK, CDK2, Cyclin E, and Ki67 positivity. Furthermore, EVI1 knockdown inhibited migratory capacity and anchorage independent growth of human PCa cells. Noteworthy, EVI1 knockdown furthermore suppressed in vitro tumor sphere and in vivo tumor initiation capacity of human PCa cells, suggesting a role as a prostate CSC protein. Finally, EVI1 expression was induced in experimentally derived docetaxel-resistant PCa cells and knockdown of EVI1 in these cells was able to restore their sensitivity to docetaxel. Conclusion: In summary, these data indicate EVI1 as a novel gene involved in prostate PCa progression and therapy resistance that may control prostate carcinogenesis at the stem cell level. Disclosure: No conflict of interest disclosed. P885

Age and prostate cancer treatment: Results from the Cancer Registry Baden-Württemberg Friedrich S.1, Hermann S.1, Arndt V.1 Deutsches Krebsforschungszentrum, Epidemiologisches Krebsregister BadenWürttemberg, Heidelberg, Germany 1

Introduction: Prostate cancer is the most frequent incident cancer in Baden-Württemberg and affects predominantly older men. The aim of this evaluation is to give an overview of the treatment pattern in prostate cancer patients in Baden-Württemberg with a focus on age-specific differences. Method: Treatment data of prostate cancer cases (PCC) (ICD-10 C61), diagnosed 2013 and reported to the Cancer Registry Baden-Württemberg (CRBW), was evaluated. The frequency of different treatment schemes [radiotherapy (RT), operation (OP), hormone-therapy (HT) or a combination] are stratified by age group (≤69 and ≥70 years) and Gleason score (≤6 and ≥7) at time of diagnosis. Results: 6.655 newly diagnosed PCC were documented at the CRBW 2013 (UICC-Stadium n = 2.673 I: 24,2%, II: 36,4, III:20,3%, IV:19,2%). Median age was 71,2 years. According to estimations more than 90% of the expected PCC in Baden-Württemberg were reported to the CRBW. However, 47% of the PCC were reported without treatment information and for 99 men ´wait and see´ was reported. Overall, RT was reported in 12%, OP in 23%, HT in 4% and multimodal therapy in 14%. The Gleason score was documented for 83% of the PCC, of these 1.357 patients having a Gleason score ≤6 and 4.142 >=7. Stratifying data by age group showed that an OP as primary treatment was reported less frequently for older PCC (age 70+) than for younger patients (21%, 39%). Older PCC received HT (12%, 6%) or RT more often

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(16%, 9%). However, 51% (70+ years) and 45% (< 70) of PCC were reported stating ´other therapy´ or without treatment information. Stratifying the data according to Gleason score showed that a sole hormone-therapy and a combination of different treatments was reported more frequently for patients with Gleason score ≥7 than for those with Gleason score ≤6 (3,4%; 18,1%). 60% with Gleason score ≤6 and 42% with a Gleason score ≥7 were reported without treatment information. Conclusion: Our data show that primary therapy is associated with the age at diagnosis: men, older than 70 years of age, receive OP less and RT as well as HT more frequently than younger men. PCC with a Gleason score ≥ 7 at diagnosis receive more often multimodal therapy. A limiting factor is that it is currently not possible to differentiate between “not reporting” and “not receiving” therapy. For meaningful analyzes, it is essential that therapies are reported fully to the cancer registry and that “watchful waiting and active surveillance” can be reported. Disclosure: No conflict of interest disclosed. P886

Analysis of efficacy of Cabazitaxel treatment in metastatic castration resistant prostate cancer in second and later lines. Single center experience Zschäbitz S.1, Vallet S.1, Hadaschik B.2, Pahernik S.3, Duensing S.2, Spath C.1, Jäger D.1, Hohenfellner M.2, Grüllich C.1 Universitaetsklinikum Heidelberg, Med. Klinik VI, Med. Onkologie, Heidelberg, Germany, 2Universitaetsklinikum Heidelberg, Urologie, Heidelberg, Germany, 3 Klinikum Nürnberg Nord, Urologie, Nürnberg, Germany 1

Introduction: Several new treatment options for patients with metastatic castration resistant prostate cancer (mCRPC) have been approved within the last years – among them cabazitaxel (CAB), abiraterone acetate, enzalutamide, and radium-223. No factors predictive for efficacy are known and the optimal treatment sequence has yet to be defined. Methods: We analyzed all patients with mCRPC treated with CAB at our institution between 2011 and 2016. Data were retrieved retrospectively from the prospective electronical patient chart. Results: 40 patients received CAB (27.5% 2nd line, 35.0% 3rd line, 37.5% >3rd line). Median progression free survival (PFS) was 4.0 months (95%CI 2.6-5.5). Median overall survival (OS) was 12.0 months (95%CI 8.3-15.7). There were no differences in OS and PFS regarding treatment line. Duration of remission on 1st line treatment (> 6 months vs. < /= 6 months) was associated with a longer PFS with subsequent CAB treatment (5.2 months (95%CI 2.4-8.0) vs. 3.0 months (95%-CI 2.1-3.9); p = 0.02). Patients with visceral metastases had a shorter PFS (3.0 months; 95%CI 1.7-4.3) and OS (8.0 months; 95%CI 4.9-11.1) on CAB compared to patients who had bone or lymph node lesions only (PFS: 8.0 months; 95%CI 2.8-13.2; p = 0.006; OS: 25.0 months; 95%CI 10.6-39.4; p < 0.001). Conclusions: Results from our patient cohort suggest that a longer PFS to any 1st line treatment for mCRPC is correlated with a longer PFS to CAB for any later line treatment. Patients with nodal and bone metastases only had a significantly superior PFS and OS with CAB treatment than patients with visceral metastases. Disclosure: No conflict of interest disclosed.

Abstracts

P887

Effects of concomitant medication and baseline parameters on overall survival and safety in German metastatic castration-resistant prostate cancer patient subset treated with Radium-223 dichloride within the international early access program (iEAP) Heidenreich A.1, Saad F.2, Carles J.3, Gillessen S.4, Heinrich D.5, Gratt J.6, Miller K.7, Nilsson S.8, O‘Sullivan J.9, Tucci M.10, Wirth M.11 University Hospital Cologne, Department of Urology, Cologne, Germany, University of Montreal Hospital Centers, Dept. of Urology, Montreal, Canada, 3 Vall d’Hebron University Hospital, Institute of Oncology, Barcelona, Spain, 4 Kantonsspital St. Gallen, Dept of Oncology, St. Gallen, Switzerland, 5Akershus University Hospital, Dept. of Oncology, Lørenskog, Norway, 6Bayer Healthcare Pharmaceuticals, Whippany, United States, 7Charité University Medicine Berlin, Dept. of Urology, Berlin, Germany, 8Karolinska University Hospital, Dept. of Oncology, Stockholm, Sweden, 9Queen’s University Belfast and the Northern Ireland Cancer Centre, Centre for Cancer Research and Cell Biology, Belfast, United Kingdom, 10San Luigi Hospital, Dept. of Oncology, Orbassano, Italy, 11 University Hospital Carl Gustav Carus, Dept. of Urology, Dresden, Germany 1 2

Background: Results from the pivotal ALSYMPCA study reported an improved overall survival (OS) in mCRPC patients (pts) with symptomatic bone lesions treated with radium-223 (Ra-223) vs placebo (median 14.9 vs 11.6 months [mos], HR = 0.70). These results were supported by a phase IIIb trial (iEAP) including data from 696 pts recruited from 14 countries previously presented at ECC 2015 (1). Here we present the data of the German patient subset. Methods: Patients received Ra-223, 50 kBq/kg (iv injection) every 4 weeks for 6 cycles. Primary endpoints were safety and OS. The effects of concomitant medications and baseline (BL) factors on OS were assessed. Results: 124 German pts were treated; 61% received all 6 Ra-223 injections. At BL: median age was 69.4 years; 91% of pts were ECOG PS 0-1; pain was reported as: no pain, mild/moderate, and severe in 26%, 58%, and 14% respectively. 56% of pts received prior therapy with docetaxel. For pts treated with concomitant therapy: 28% received abiraterone (n = 35); 18% received denosumab (n = 23); 32% had bisphosphonates (n = 40) and 12% received enzalutamide (n = 15). Grade 3/4 AEs were reported in 32% of pts; 23% discontinued Ra-223 due to AEs. At the time of analysis median OS was 13 mos. In patients without prior use of docetaxel median OS was not evaluable vs. 10 mos for patients with prior use of docetaxel. OS appeared to be longer in those patients with the following BL characteristics: ALP < 220 U/L vs ≥ 220 U/L; ECOG PS 0-1 vs ≥ 2; no pain vs mild-moderate vs severe; and in those who received concomitant abiraterone/enzalutamide. Conclusions: Results from German patient population are similar to the data of the whole study population. References: 1 O’Sullivan et al., ECC 2015, Abstract No. 2561. Disclosure: Axel Heidenreich: Advisory Role: Astellas Pharma; Bayer; Janssen; Sanofi; Financing of Scientific Research: Amgen; Astellas Pharma; Bayer; Dendreon; Ferring; Ipsen; Janssen; Pfizer; Sanofi; Takeda; Expert Testimony: Astellas Pharma; Bayer; Sanofi Manfred Wirth: Advisory Role: Bayer; Dendreon; Ferring; Janssen; Merck; Takeda; Financing of Scientific Research: Apogepha; Astellas Pharma;Orion Pharma; Sanofi; Expert Testimony: Apogepha (Inst); Ferring (Inst); Sanofi (Inst); Takeda (Inst)

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Nivolumab monotherapy in metastatic urothelial cancer (mUC): efficacy (by PD-L1 expression) and safety results from the CheckMate 032 study Jaeger D.1, Sharma P.2, Bono P.3, Kim J.4, Spiliopoulou P.5, Calvo E.6, Pillai R.N.7, Ott P.A.8, de Braud F.9, Morse M.10, Le D.11, Chan E.12, Harbison C.13, Lin C.-S.13, Tschaika M.13, Azrilevich A.13, Rosenberg J.14 Heidelberg University Hospital, Heidelberg, Germany, 2MD Anderson Cancer Center, University of Texas, Houston, United States, 3Comprehensive Cancer Center, Helsinki University Hospital, Helsinki, Finland, 4Yale Cancer Center, New Haven, United States, 5Beatson West of Scotland Cancer Centre, Glasgow, United Kingdom, 6START Madrid, Centro Integral Oncológico Clara Campal, Madrid, Spain, 7Emory Winship Cancer Institute, Atlanta, United States, 8DanaFarber Cancer Center, Boston, United States, 9Istituto Nazionale dei Tumori, Milan, Italy, 10Duke University Medical Center, Durham, United States, 11Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, United States, 12Vanderbilt University, Nashville, United States, 13Bristol-Myers Squibb, Princeton, United States, 14Memorial Sloan Kettering Cancer Center, New York, United States 1

Introduction: Nivolumab, a fully human IgG4 programmed death-1 immune checkpoint inhibitor antibody, has recently demonstrated promising efficacy and acceptable safety in an open-label, multicenter phase I/II study in unselected patients with mUC after ≥1 prior lines of platinum-based therapies (NCT01928394). Here, we report efficacy by differing levels of programmed death ligand 1 (PD-L1) expression and safety of nivolumab. Methods: Patients with mUC, unselected by PD-L1 expression status, received nivolumab 3 mg/kg intravenously every 2 weeks until progression or discontinuation. Patients were allowed to continue treatment beyond progression and to cross over to nivolumab combined with ipilimumab when they met prespecified protocol criteria. Tumor PD-L1 membrane expression was assessed with Dako PD-L1 immunohistochemical staining. Primary endpoint: objective response rate (ORR; RECIST 1.1); other endpoints: safety, progression-free survival (PFS), overall survival (OS), and duration of response (DOR). Results: Of 78 treated patients (median age 65.5 years; range, 31-85), 51 had received ≥2 prior therapies. At a median follow-up of 213 days (range, 22-499), 33.3% of patients remained on therapy; treatment discontinuation was mainly due to disease progression. Median number of doses was 8.5 (range, 1-34); 70.5% received >4 doses. PD-L1 was evaluable in 66 patients (85%). Overall efficacy results are shown in the table. In patients with PD-L1 expression ≥1% (n = 25) vs those with < 1% (n = 42), ORR (95% confidence interval [CI]) was 24% (9.4-45.1) vs 26.8% (14.2-42.9); median (95% CI) PFS, 5.5 months (1.4-not estimable [NE]) vs 2.8 months (1.5-6.5). Grade 3 or 4 treatment-related adverse events (TRAEs) occurred in 20.5% of patients; most frequent were ↑ lipase, ↑ amylase (3.8% each), and fatigue, ↓ neutrophils, and dyspnea (2.6% each). Grade 5 TRAEs occurred in 2.6% of patients (pneumonitis [n = 1] and thrombocytopenia [TTP; n = 1]). No grade 3 or 4 pneumonitis or TTP was reported. Conclusions: Nivolumab monotherapy showed encouraging efficacy and acceptable safety regardless of PD-L1 expression in previously treated, unselected patients with mUC. Tab. 1.

Parameter ORR (confirmed), % (95% CI) Median PFS, months (95% CI) Median OS, months (95% CI) Median time to response, months (SD) Median DOR, months (95% CI)

Nivolumab (n = 78) 24.4 (15.3−35.4) 2.8 (1.5−5.5) NE (7.0−NE) 1.5 (2.1) NE (5.5–NE)

Disclosure: Dirk Jaeger: Advisory Role: Roche, BMS Bayer Jonathan Rosenberg: Expert Testimony: Genentech, Oncogenex, Agensys, Mirati Therapeutics

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Posterdiskussion

Tumorbiologie, Zellbiologie 1 P889

Functional upstream open reading frame – encoded peptides in human proto-oncogenes Wethmar K.1,2, Schulz J.2, Leutz A.2,3 University Hospital Muenster, Department of Medicine A, Hematology, Oncology and Pneumology, Muenster, Germany, 2Max-Delbrueck-Center for Molecular Medicine, Department of Cell Differentiation and Tumorigenesis, Berlin, Germany, 3Humboldt-University, Department of Biology, Berlin, Germany 1

Human transcript leader sequences (TLSs) contain approximately 26.500 translation initiation codons (uAUGs) preceding the canonical main protein coding sequences (CDSs). Numerous uAUGs serve to initiate upstream open reading frames (uORFs) that may be translated into small peptides (uPeptides). Due to the predominantly small size and rapid turnover, only a minor fraction of endogenous uPeptides has been detected by current mass spectrometric approaches. The translation of uORFs is generally associated with repressed protein production from related downstream CDSs, yet the function of uORF-encoded peptides is mostly unknown. Here, we investigated the expression, localization and translational regulatory impact of uPeptides for a set of human proto-oncogenes. We readily detected uPeptide expression for transcripts of HCK, MAP2K3, KDR, MET and MDM2 using a uPeptide expression system that permitted insertion of 5´-partial endogenous TLSs. Immunofluorescent staining of HA-tagged uPeptides for HCK, MAP2K3 and KDR demonstrated a mostly ubiquitous cellular distribution, yet occasionally, (peri)-nuclear localization was detected for HCK and MAP2K3 uPeptides, suggesting a potential trans-regulatory role. Several uPeptides were selected to investigate the effect of experimentally introduced alterations in their amino acid sequences. Mutant versions of the MAP2K3, HCK and MDM2 uPeptides showed enhanced translation of the associated downstream protein coding regions. Furthermore, sequence alterations caused changes in the expression level of the HCK uPeptide itself, underlining the importance of conserved amino acid residues for the peptide. We conclude that uPeptides are readily detectable and functional in a number of proto-oncogenes. Our data imply that naturally occurring mutations in conserved uPeptide coding sequences may cause changes in both, the abundance of uPeptides and associated downstream proteins. Previous studies in yeast and plants support the notion that uPeptides can serve as translational `pepto-switches´ controlling downstream protein synthesis in response to various ligands. Accordingly, our evidence for the translation of functional uPeptides from proto-oncogenic transcripts inspires the idea that uPeptides may be pharmacologically targetable to enable drug-mediated regulation of gene expression in cancer therapy. Disclosure: No conflict of interest disclosed. P890

Characterization of CD98hc in pancreatic ductal adenocarcinoma Bianconi D.1, Herac M.2, Gleiss A.3, Unseld M.1, Weigl R.1, Schindl M.2, Scheithauer W.1, Zielinski C.1, Prager G.1 Medizinische Universität Wien, Innere Medizin I, Vienna, Austria, 2Klinisches Institut für Pathologie, Medizinische Universität Wien, Vienna, Austria, 3Institut für Klinische Biometrie, CeMSIIS, Medizinische Universität Wien, Vienna, Austria 1

Introduction: Despite recent advances in medical oncology, pancreatic ductal adenocarcinoma (PDAC) represents one of the most aggressive cancers with limited prognosis. With less than 5% five-year-overall survival, the characterization of new therapeutic targets is urgently needed. CD98hc, a transmembrane type II protein, is expressed in some tumor cells and therefore, it represents a promising novel therapeutic target.

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Thus, the aim of the study is to characterize the putative malignant role of CD98hc in PDAC. Methods: CD98hc protein expression level was analyzed in human pancreatic cancer tissue (222 cases) and matched adjacent tissue (141 cases) via immunohistochemistry. Additionally, pancreatic cancer cells were isolated from fresh human tumor tissue and CD98hc positive cells were phenotypically analyzed via flow cytometry. To characterize the functional role of CD98hc in PDAC, CD98hc was downregulated in the pancreatic cancer cell lines BxCP3 and PANC1 by lentiviral transduction. Cell proliferation, cell cycle progression and tumorsphere formation capacity were analyzed in vitro. Results: Using immunohistochemistry, we demonstrated that CD98hc is aberrantly expressed in human pancreatic cancer tissue and matched adjacent tissue. However, there was no significant difference in CD98hc expression in the different grades and stages. Although Kaplan-Meier survival curves and Cox analyses did not revealed a significant association between CD98hc expression and overall survival, we observed a trend that suggests a possible association between absence of CD98hc expression and prolonged survival. Additionally, our results showed that CD98hc positive cells isolated from fresh tumor tissue co-expressed other known markers of PDAC such as MUC4 and MUC1.To determine the role of CD98hc in PDAC, cell behavior between CD98h positive cells and CD98hc low expressing cells was compared. We found that CD98hc downregulation significantly inhibited tumorsphere formation and cell proliferation by arresting cell cycle. Conclusion: In this study, we demonstrated that CD98hc is overexpressed in resected tumor tissue as well as in matched adjacent tissue of patients with pancreatic cancer. Moreover, our data suggests that CD98hc expression increases tumorigenesis by enhancing cell proliferation and promoting anchorage-independent growth in vitro. Although further studies are needed, our results suggest that CD98hc might be a novel therapeutic target of PDAC. Disclosure: No conflict of interest disclosed. P891

Targeting mitotic exit for cancer treatment Greil C.1, Schnerch D.1, Felthaus J.1, Wider D.1, Schüler J.2, Duyster J.1, Engelhardt M.1, Wäsch R.1 Uniklinik Freiburg, Freiburg, Germany, 2Oncotest, Freiburg, Germany

Introduction: Targeting mitosis of cancer cells with spindle assembly checkpoint (SAC) activating agents is a common therapeutical approach. In different tumors residual anaphase-promoting complex/cyclosome (APC/C) acitivity and continuous background cyclin B degradation may lead to mitotic exit and cells may survive antimitotic treatment. Targeting the APC/C or its activator Cdc20 can prevent mitotic slippage. Therefore, inhibition of cyclin B degradation via APC/C inhibition in cells treated with antimitotic drugs should lead to more efficient tumor cell eradication. Proteasome inhibitors indirectly reducing cyclin B degradation may have a similar effect as APC/C-inhibitors in this mechanistic context. Methods: Cell lines and primary tumor cells from different neoplasms cultivated according to established protocols were treated with combinations of antimitotic agents or the Plk1-inhibitor volasertib and the proteasome inhibitor bortezomib or the APC/C-inhibitors proTAME/Apcin. FACS analysis was conducted to determine G2/M-arrest and apoptotic rate, soft agar colony assays were used to evaluate cell growth and determine lasting tumor eradication. A newly established xenotransplant AML mouse model was treated with the different combinations and tumor growths and survival were analyzed. Results: Sequential treatment with paclitaxel and bortezomib was equally effective as the combination of paclitaxel with proTAME/Apcin in HeLa cells. The combination of paclitaxel with bortezomib was more effective than treatment with paclitaxel alone in patient-derived NSCLC cells leading to higher rates of G2/M-arrest and reduced cell growth. Combined treatment with volasertib and bortezomib significantly enhanced the mitotic block and subsequent mitosis in Kasumi-1 cells and primary AML

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samples. Expression of the anti-apoptotic Bcl2-family member Mcl1 was reduced in AML cell lines. Its overexpression enhanced G2/M-block and reduced apoptosis following treatment with volasertib and bortezomib. Combined treatment with bortezomib and volasertib prolonged survival in our xenograft AML mouse model. Conclusions: The most efficient anti-proliferative effect due to mitotic arrest may be obtained by combining SAC-activating agents and agents blocking cyclin B proteolysis using low doses achievable in-vivo. Our data provide a strong rationale for clinical phase I/II trials with approved drugs as well as experimental substances to further exploit this highly promising concept. Disclosure: No conflict of interest disclosed. P892

Impaired and delayed response of neutrophils of immunosuppressed HSCT patients against Aspergillus fumigatus conidia Jahreis S.1,2, Hartung S.1,2,3, Rauh C.1,2, Wagner K.1,2,3, Hilgendorf I.1, Rummler S.4, Hochhaus A.1, von Lilienfeld-Toal M.1,2,3 Universitätsklinikum Jena, Klinik für Innere Medizin II, Jena, Germany, 2LeibnizInstitut für Naturstoff-Forschung und Infektionsbiologie, Infektionen in der Hämatologie und Onkologie, Jena, Germany, 3Universitätsklinikum Jena, Integriertes Forschungs- und Behandlungszentrum Sepsis und Sepsisfolgen, Jena, Germany, 4Universitätsklinikum Jena, Institut für Transfusionsmedizin, Jena, Germany 1

Introduction: The opportunistic human pathogen Aspergillus fumigatus is the most important cause of fatal fungal infections in immunocompromised individuals, such as chemotherapy patients or solid organ transplant recipients. Generally, infection is established by fungal conidia which are ubiquitiously present in the air. Phagocytes, i.e. monocytes and neutrophil granulocytes, are in the first line of defense against invading fungi. We aim at elucidating potentially different responses of human phagocytes towards A. fumigatus from immunosuppressed patients versus healthy controls. Methods: Leukocytes from healthy blood donors or from immunosuppressed hematopoietic stem cell transplantation (HSCT) patients were co-incubated with resting or pre-swollen FITC-labelled A. fumigatus conidia. Leukocytes without conidia served as control. Afterwards, cells were analysed by flow cytometry for phagocytosis as well as neutrophil marker expression. Results: Up to 63% of neutrophils from healthy donors performed phagocytosis of A.fumigatus conidia after 2 and 4 hours. Patient-derived neutrophils revealed a massive impairment of phagocytosis (35% or lower) at all time points. Cell activation, measured by the neutrophil-specific degranulation marker CD66b, was significantly lower in patients than in healthy samples. In addition, pro-inflammatory IL-1β release from patient leukocytes could be only detected at a concentration of 4.1 (± 4.0) pg/ml compared to 19.8 (± 12.2) pg/ml in healthy individuals using ELISA. Cell adhesion molecules CD11b and CD62L were dysregulated in immunosuppressed neutrophils. In healthy neutrophils, CD11b was upregulated after 0.5 hours of co-incubation compared to conidia-free controls and returned to basal levels within 4 hours. By contrast, CD11b expression in patient-derived neutrophils remained at elevated levels after an initial increase. CD62L expression in immunosuppressed samples was significantly higher than in healthy samples at basal levels as well as after co-incubation. Conclusion: Dysregulation of CD11b and CD62L adhesion molecule expression in patient neutrophils compared to healthy individuals may hint at impaired migration abilities. This would explain the decreased phagocytosis rate and at the same time suggest a general delay of the response of neutrophils against A. fumigatus conidia in immunosuppressed patients. Disclosure: Susanne Jahreis: No conflict of interest disclosed. Marie von Lilienfeld-Toal: Advisory Role: MSD, Celgene; Financing of Scientific Research: MSD, Celgene, Janssen Cilag, Gilead; Expert Testimony: MSD, Pfizer; Other Financial Relationships: Celgene, Janssen Cilag, Gilead, Astellas

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B cells derived from patients with von Hippel-Lindau (VHL) disease display impaired antigen presentation upon CD40 activation Becker H.J.1, Reuter S.1, Garcia Marquez M.1, Schlößer H.A.1,2, von BergweltBaildon M.1, Theurich S.1 Uniklinik Köln, Klinik I für Innere Medizin, Interventionelle Immunologie, Köln, Germany, 2Uniklinik Köln, Allgemein-, Visceral- und Tumorchirurgie, Köln, Germany 1

Introduction: Germline mutations in the von Hippel Lindau (VHL) gene result in VHL disease, which puts patients at high risk for developing characteristic neoplasias such as pheochromocytomas, hemangioblastomas, and renal cell carcinomas. Molecularly, the VHL protein plays a pivotal role in the VHL/HIF axis, disruption of which has been reported to contribute to tumor progression via immunomodulatory mechanisms. However, most reports have focused on T cell function and the tumor microenvironment. Using CD40 activated B cells (CD40B) as an in vitro model for B cell activation, we aimed at elucidating phenotypical and functional characteristics of activated B cells in this patient collective. Methods: Primary blood cells from 15 individuals with VHL disease were obtained. B cells were isolated and activated with soluble CD40L. Immune phenotyping was carried out using flow cytometry, and phosphorylation profiles of CD40 signaling pathway molecules were analyzed with the phosphoflow assay. 5-day allogenic mixed lymphocyte reactions (MLR) were performed with CD40B cells as antigen presenting cells and CD3+ T cells as responder cells. Results: After 4 days of CD40 stimulation, expression of co-stimulatory molecules CD80 and CD86 in VHL B cells was attenuated by 46% and 59%, respectively. Hypophosphorylation of CD40 signaling proteins pAKT, pERK1/2, pNFkB, and pp38 was evident in VHL B cells when compared to healthy controls. In allogenic mixed lymphocyte reaction experiments, VHL CD40B cells triggered significantly less T cell activation and proliferation in both CD4+ (63%, 28%) and CD8+ (56%, 28% at B:T ratios 1:1, 1:5, respectively; relative to healthy CD40B cells) subsets. No association between the degree of phenotypical and functional alteration and clinical VHL subtype was observed. Conclusions: Our findings suggest that mutations in the VHL gene confer a significantly less immunostimulatory phenotype to B cells upon CD40-mediated activation. Impaired immune function of these cells might contribute to the increased incidence of neoplasms in VHL disease patients. Further investigation focusing on this patient collective might provide new aspects to tumorigenesis and immunotherapy. Disclosure: No conflict of interest disclosed. P894

Exploiting proteasome function in senescence-associated proteotoxicity as target principle in lymphoma therapy Anell Rendon D.1, Schmitt C.A.1,2 Max-Delbrück-Center for Molecular Medicine, Berlin, Germany, 2Charité – Universitätsmedizin Berlin (CVK), Hematology, Oncology, and Tumor Immunology, Berlin, Germany 1

Cellular senescence is a cell-cycle arrest fail-safe mechanism triggered by stress signals like oncogene activation and DNA damage induced by chemotherapy. Previously, we were able to show that cells undergoing therapy-induced senescence (TIS) undergo massive proteotoxic stress as a consequence of the increased protein production associated with the senescence-associated secretory phenotype (SASP). Although considered to be terminal, tumor cells locked into senescence may be harmful via their largely pro-inflammatory secretome, or might be of particular biological aggressivity if they would manage to re-enter the cycle. Accordingly, we already demonstrated that therapeutic strategies that secondarily eliminate drug-senescent cells extend long-term survival after lymphoma chemotherapy in pre-clinical models.

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Here, we address the role of protein degradation by the proteasome in senescent vs. non-senescent aggressive B-cell lymphoma cells. We now present that drug-senescent Eµ-myc transgenic, Bcl2-expressing mouse lymphoma cells display, as previously shown for autophagy inhibition, enhanced vulnerability to proteasome blockade in vitro compared to their equally treated but genetically senescence-impaired counterparts. Importantly, when we simultaneously inhibited proteasome and autophagy activity with moderate doses of both agents, TIS lymphomas exhibited additive susceptibility and rapid loss of viability. Adding to our previous findings that senescent cells rely on a hypercatabolic metabolic state to fuel energy-consuming autophagy as a mechanism to cope with toxic senescence-associated peptides, we now demonstrate a similar dependency with respect to protein degradation by an intact proteasomal degradation machinery. Our therapeutic approach converts the rather detrimental implications of largely pro-inflammatory SASP factors into a novel strategy to eliminate malignant cells in a synthetically lethal manner. Disclosure: No conflict of interest disclosed. P895

MTA (5´-deoxy-5´-methylthioadenosine) suppresses function of human mDC and T cells and represents a mechanism of tumor immune escape Strobl C.1, Henrich F.1, Peter K.2, Singer K.2, Kreutz M.2, Kremer A.1, Mackensen A.1, Aigner M.1 University Hospital of Erlangen-Nuremberg, Department of Internal Medicine 5 - Hematology/Oncology, Erlangen, Germany, 2University Hospital Regensburg, Deptartment of Internal Medicine 3 - Hematology/Oncology, Regensburg, Germany 1

Creation of an immunosuppressive microenvironment is one mechanism of tumor immune escape. Amongst others, tumors can take advantage of their metabolic alterations to inhibit immune responses. In recent years, methylthioadenosine phosphorylase (MTAP) deficiency gained more and more interest as putative immuno-inhibitory metabolic dysregulation. Loss of MTAP is found in various tumor entities and leads to accumulation of 5’-deoxy-5’-methylthioadenosine (MTA). Lack of MTAP is associated with an inferior response towards adjuvant interferon-α therapy and a higher risk for metastatic disease in malignant melanoma. Here we examined the role of tumor-secreted MTA in suppression of anti-tumor T cell responses. We were able to show that MTA has immuno-inhibitory functions upon human T cell proliferation, viability, activation, differentiation, clonal expansion and effector functions. We therefore sought to use retroviral transduction to generate MTAP-overexpressing T cells as a putative strategy to overcome MTA-mediated inhibitory effects. In addition, we investigated the effect of MTA upon the differentiation and maturation of human monocyte derived mDC. We successfully generated stable MTAP-overexpressing primary human T cell as well as T cell lines. Experiments revealed influence of MTAP overexpression on T cell survival and proliferation when co-cultured with MTA compared to mock control suggesting a promising approach to reconstitute inhibition by MTA. In addition we found that presence of MTA also interferes with DC maturation and their potential to induce cytokine secretion from T cells. Our data emphasizes the importance of tumor metabolites such as MTA in the tumor milieu and provides a strategy of tumor immune escape. Further characterization of the molecular mechanisms of MTA-induced immunosuppression will help to develop more effective immune-based therapies against MTAP-deficient tumors. Disclosure: No conflict of interest disclosed.

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Autonomous processing of microRNA in exosomes produced by chemoresistant colon cancer cells in vitro Rösinger S.1, Kalipciyan M.1, Steger G.G.1, Mader R.M.1 Univ.Klinik für Innere Medizin I, Klinische Abteilung für Onkologie, Wien, Austria 1

Introduction: microRNA (miR) are small non-coding RNA, which control gene expression by targeting mRNA. When deregulated, microRNA play functional roles in the pathophysiology of malignant processes. Remarkably, microRNA are selectively transferred from the cancer cell into exosomes. Circulating in the organism, these microvesicles play an important role in cell-to-cell communication. In this study, we investigated the expression of microRNA processing enzymes and their functionality in exosomes obtained from three different chemoresistant cancer cells. Methods: As a model, we used colon cancer cell lines with progressing stages of chemoresistance against 5-fluorouracil (FU): SW480 (primary adenocarcinoma), SW620 (lymph node metastasis) and three resistant subclones of SW620. From isolated exosomes, the expression of microRNA processing enzymes was analysed by immunoblotting and the processing of miR-precursors to mature microRNA by qPCR. Results: In our chemoresistant model, several microRNA processing enzymes were detected in exosomes from cancer cells with different degrees of resistance, some of them differentially regulated. As an example, there was an overexpression the enzymes exportin-5 (XPO5), Argonaute-2 (AGO2) and DGCR8 in the low resistant phenotype (resistant against 5 µM FU). To test the functionality of these exosomal enzymes, precursor of miR-4653-5p and miR-4653-3p were transfected into exosomes. After 48 hours, an almost quantitative maturation of both microRNA could be detected by qPCR, whereas the transfected precursor was below 5% of the baseline value. This exosomal processing was independent of the degree of chemoresistance of the parent cell line. Conclusions: Exosomes effectively process microRNA as shown for miR4653, which was hardly detected in our chemoresistant colon system at baseline. The differential expression of microRNA processing enzymes in exosomes did not impact on the conversion of pre-miR into mature microRNA, which was independent of the degree of chemoresistance. These data strongly emphasise the autonomous character of circulating exosomes in cancer with the ability not only to transport information from cell to cell, but also to effectively process information based on microRNA. Disclosure: No conflict of interest disclosed. P897

Comparison of phenotype and gene expression analysis of circulating epithelial tumor cells (CETCs) and tumorspheres in colorectal cancer patients Schott D.1, Pizon M.1, Pachmann U.1, Pachmann K.1 Transfusionsmedizinisches Zentrum Bayreuth, Bayreuth, Germany

Background: Colorectal cancer is one of the most commonly diagnosed and lethal cancers worldwide. Tumor recurrence and metastasis are two critical survival-influencing factors. Circulating cancer stem cells are a rare subpopulation of tumor cells circulating in the blood of cancer patients. They are suspected to be responsible for tumor growth, metastasis, resistance towards therapy and thus progression. Culture of tumorspheres has been used to detect cancer stem cells from primary tumor material or cell lines. In contrast, we established an effective method for identification of cancer stem cells from circulating tumor cells in the blood of patients with colorectal cancer with subsequent phenotypic and genotypic characterization. Methods: Metastatic and non-metastatic colorectal cancer patients with detectable CETCs were included in the present study. CETCs were enumerated and subsequently cultured under conditions favoring growth of

Abstracts

epithelial tumorspheres. Immunofluorescence and qRT-PCR were applied to examine the metastatic ability of tumorspheres in vitro. Results: Only a small fraction of CETCs had the ability to form tumorspheres. Patients with metastatic disease had more tumor spheres, than non-metastatic patients (median 15 vs 48 spheres per 100 µl blood) indicating that the number of tumor spheres is dependent on stage of disease. Analysis of surface markers showed high levels of EpCAM and CD 133 expression. Furthermore, spheres had high enzymatic activity for ALDH 1. Array qRT-PCR analysis revealed that tumorspheres express genes of pluripotency such as SOX2, OCT4 and NANOG. There was no sphere formation in 50 healthy subjects. Conclusion: This study demonstrates that tumor stem cells are present in peripheral blood from metastatic and non-metastatic colorectal cancer patients. A more complete understanding of the biology of cancer stem cells will translate into the development of better chemotherapeutic and biological agents for the treatment of colorectal cancer. Disclosure: No conflict of interest disclosed. P898

Extracellular vesicles in plasma and urine of patients and healthy donors: loss of TFPI and increased tenase activity after kidney passage Freund A.1, Plattfaut C.1, Quecke T.1, Riemekasten G.2, Haas C.3, Gieseler F.1 1 University of Lübeck, Experimental Onkology, Lübeck, Germany, 2University of Lübeck, Department of Rheumatology, Lübeck, Germany, 3University of Lübeck, Experimental Nephrology, Lübeck, Germany

Background and aims: Extracellular vesicles (EVs) released by tumors and inflammatory cells seem to play a crucial role in tumor-associated thrombosis and metastasis. Increased levels have been found in several inflammatory diseases as well as advanced tumor stages. Based on their biogenesis, size and structure, EVs are classified into exosomes, ectosomes or microparticles (MPs), and apoptotic bodies. We compared EVs from plasma and urine (pre- and post-kidney passage) from healthy donors and patients. EV-populations were characterized by size and functional activities such as tissue-factor-related tenase activities, association of tissue factor pathway inhibitor (TFPI), and their potency to induce tumor cell migration. As we have described before, TF/FXa complexes are able to induce tumor cell migration via PAR/ERK-activation, whereas TFPI is the most potent physiological TF/FXa inhibitor. Methods: EVs were isolated using sequential centrifugation steps (2 x 2,500 x g), to accumulate EVs and reduce cell membrane fragments and larger apoptotic bodies. Phosphatidylserine (PS)-presenting MPs were captured with annexin V-coated magnetic beads as described before. EVs were characterized by size using high-resolution flow cytometry after calibration with “Megamix”. Tenase- and TFPI-activity, as well as PAR-activating potency, were examined by ELISA-based assays. For EV-induced tumor cell migration, the “Oris Migration Assay” was used. Results: EVs could be isolated as well from plasma as from urine and were characterized as exosomes, MPs and apoptotic bodies. TFPI was found to be associated with plasma EVs (pre-kidney passage), but not to the urinary EVs (post-kidney passage). Consequently, urinary EVs had significantly higher tenase activities than those from plasma. PS-presenting MPs, as captured by annexin-coated magnetic beads, were found to be the major fraction to induce tumor cell migration through the PAR/ERK pathway. Conclusion: EVs can be isolated and characterized as well from plasma as from urine. During kidney passage, EVs loose their TFPI-association resulting in significantly higher tenase activity as well as PAR-activating and migration-inducing potency. Urinary EVs are an additional source of material and are well suited for investigating the possible patho-physiological and diagnostic relevance of EVs released by tumor cells and inflammatory cells. Disclosure: No conflict of interest disclosed.

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Bone morphogenetic protein 4 affects invasive tumor cell subpopulation Mihajlović J.1, Fischer M.1, Hochhaus A.1, Clement J.1 Universitätsklinikum Jena, Hämatologie und internistische Onkologie, Jena, Germany 1

Background: Bone morphogenetic proteins (BMPs) are multifunctional growth and differentiation factors that play essential roles in embryo and adult for tissue development and homeostasis. Numerous studies describe divergent roles of BMPs in cancer development. It remains still unclear whether they have uni- or bi-directional role in pathology of cancer. With this study we aim to investigate the role of BMP-4 on the formation and maintenance of the metastatic cell phenotype in cancer cell lines. Methods: The study included a hepatocellular carcinoma cell line HepG2, and two breast carcinoma cell lines MCF-7 and BT-20. The cells were treated with 100ng/ml BMP-4 and 5µM dorsomorphin under standard incubation conditions. Treatment-induced changes were observed at 10, 30, 60 and 180 minutes, and 48, 72 and 96 hours following the incubation. Phosphorylation of receptor-regulated SMADs 1/5/8(9) (R-SMADs) after the treatment was detected using immunofluorescence-immunocytochemistry and Western blot. The influence of BMP-4 on cell viability and the expression of epithelial (EpCAM) and stem cell (CD44, ALDH1) markers was analyzed via flow cytometry. Results: BMP-4 induced R-SMADs’ phosphorylation in all investigated cell lines, confirming their responsiveness. The phosphorylation event was visible shortly after the incubation with BMP-4 and, depending on the cell line, persisted for at least 24 hours. The presence of dorsomorphin inhibited R-SMADs’ phosphorylation completely. The subpopulation of cells with the highest metastatic potential is supposed to have stem cell-like characteristics and an intermediate epithelial-mesenchymal phenotype. This subpopulation, defined as CD44+/ALDHhi, was reduced by BMP-4 in HepG2 and MCF-7, but not in BT-20 cells. 96 hours following BMP-4 treatment CD44+/ALDHhi cell count in HepG2 and MCF-7 was decreased to 12% and 20% of the control, respectively. However, the BMP-4 treatment did not affect the viability of investigated cells. Conclusion: BMP-4 has a negative effect on CD44+/ALDHhi subpopulation in a cell line- and time-dependent manner. This reduction of the cell subpopulation over time may be explained by intracellular processes driving the cells towards a more differentiated phenotype. Acknowledgement: First author has a scholarship from the IZKF Jena. Disclosure: No conflict of interest disclosed. P900

Multicellular spheroids: A model for nanoparticle-tumor cell interaction studies Demut J.1, Gräfe C.1, Hochhaus A.1, Clement J.H.1 Universitätsklinikum Jena, Klinik für Innere Medizin II, Abteilung Hämatologie & Internistische Onkologie, Jena, Germany 1

Introduction: Nanomaterials are increasingly used for clinical and biomedical applications as for contrast agents in magnetic resonance imaging. Especially superparamagnetic iron oxide nanoparticles (SPIONs) are developed for this purpose. The penetration into tissue and the interaction with cells are of particular interest. Multicellular spheroids (MCS) represent a model to investigate these effects on tissue-like cell structures and display an advanced in vitro test system. This study aims to compare MCS of the epithelial breast cancer cell line BT-20 with such consisting of human brain microvascular endothelial cells (HBMEC) regarding their applicability on nanoparticle-cell interaction studies. Methods: MCS composed of BT-20 cells and HBMEC were prepared with the IncuCyte Kinetic 3D Spheroid Assay and the hanging-drop method. The latter were incubated with SPIONs coated with different polymers (positive polyethylenimine, neutral starch and negative carboxymethyldextran). Their interaction with the MCS was investigated via laser scan-

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ning microscopy. To analyse the penetration depth of the SPIONs into the MCS two methods were utilized, in particular serial trypsination and histological staining upon sectioning of the spheroids. Results: There is a distinct difference between the phenotypes of both types of multicellular spheroids. BT-20 form dense packed cell aggregates, whereas HBMEC spheroids exhibit gaps in between. Interaction studies show a charge dependent patterning. Cationic particles interact strongly; neutral particles show moderate and anionic particles almost no interaction with the MCS. Serial trypsination was applied to investigate the distribution of the nanoparticles within MCS. In our hands the procedure was not suitable for accurate and reproducible preparation of cell layers. This issue could be overcome by applying embedding and sectioning of the MCS. The results reveal similar charge dependent effects as observed via the interaction studies. Furthermore the observed penetration depth differs between BT-20 and HBMEC spheroids. In case of the HBMEC MCS nanoparticles were found in deeper layers attributed to the loose package of the cells. Conclusion: Multicellular spheroids as a complex cell culture system are suitable to investigate the interaction and penetration of SPIONs with/ into tissue. This work was supported in part by the DFG high priority program 1681 (Grant CL202/3-1) and the BMBF joint project NanoBEL (grant 03XP0003E). Disclosure: No conflict of interest disclosed. P901

Influence of Aspergillus fumigatus on natural killer cells of healthy subjects and patients with acute myeloid leukemia Ritter E.1,2, Hartung S.1,2,3, Jahreis S.1,2, Rummler S.4, Hochhaus A.1, von Lilienfeld-Toal M.1,2,3 Klinik für Innere Medizin II, Universitätsklinikum Jena, Hämatologie und Internistische Onkologie, Jena, Germany, 2Leibniz-Institut für NaturstoffForschung und Infektionsbiologie, Hans-Knöll-Institut, Jena, Germany, 3 Integriertes Forschungs- und Behandlungszentrum Sepsis und Sepsisfolgen (CSCC), Jena, Germany, 4Institut für Transfusionsmedizin, Universitätsklinikum Jena, Jena, Germany 1

Introduction: Invasive fungal disease (IFD) is a major threat for patients with immunosuppression after myelosuppressive chemotherapy. In the majority of cases, IFD results from infections with the mold Aspergillus fumigatus (A. fumigatus). Previous data point to an involvement of natural killer (NK) cells in the innate immune response to molds. Currently, however, the immune response of NK cells to A. fumigatus remains incompletely understood and it is not clear, how the mold affects the effector cell. In the present study, we aim at elucidating as to which extent A. fumigatus conidia modulate the expression of immunorelevant surface antigens and cytotoxicity of NK cells. Methods: Leukocytes and MACS-purified NK cells were isolated from either healthy donors (n = 10) or patients (n = 5) with acute myeloid leukemia after chemotherapy and regeneration of blood cell counts and co-incubated with either swollen A. fumigatus conidia or supernatant of swollen A. fumigatus conidia for 2 hours. Incubation in cell culture medium only served as control. Quantification of surface antigen expression in CD3-CD56+ NK cells was performed using flow cytometry in both the leukocyte and purified NK cell model. A CD107a mobilization assay was used to assess the effect of A. fumigatus conidia on NK cell cytotoxicity against K562 tumor cells (healthy donors: n = 5, patients: n = 5). Results: For healthy subjects, co-incubation with swollen A. fumigatus conidia resulted in a significant reduction in the fraction of NK cells expressing the natural cytotoxicity receptors (NCRs) NKp30, NKp44 or NKp46 or the activation antigen CD69. However, NK cell cytotoxicity (CD107a+) against the tumor cell line K562 was not suppressed by conidia. Additionally, co-incubation with supernatant significantly reduced the expression of NKp46, but not the other NCRs, in the leukocyte model. Of note, patient-derived NK cells exhibited a more pronounced expression of NCRs in general. A reduction of NKp30+ NK cells derived from patients could be observed after co-incubation with conidia or supernatant. In contrast

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to the observation in healthy donors, the other NCRs were not downregulated. Conclusions: In summary, our data point to an immunosuppressive effect of A. fumigatus conidia on NK cells from healthy subjects. In contrast, conidia-induced changes in the expression profile of immunorelevant surface antigens appear to be markedly reduced in NK cells from leukemic patients following chemotherapy. Disclosure: Ellen Ritter: Other Financial Relationships: Pfizer, Gilead Marie von Lilienfeld-Toal: Advisory Role: MSD, Celgene; Financing of Scientific Research: MSD, Celgene, Janssen Cilag, Gilead; Expert Testimony: MSD, Pfizer; Other Financial Relationships: Celgene, Janssen Cilag, Gilead, Astellas P902

The role of L1 retrotransposon in circulating tumor cells Apostolou P.1, Papasotiriou I.1 Research Genetic Cancer Centre Ltd (R.G.C.C. Ltd), Florina, Greece

Introduction: LINE1 (Long interspersed nuclear element 1) or L1 belongs to the family of non-long terminal repeat (non-LTR) retrotransposons. It comprises about 17% of the human genome and consists of two different open reading frames (ORFs). The ORF1 has nucleic acid chaperone activity while the ORF2 contains both endonuclease and reverse transcriptase activities. Retrotransposons can induce mutations or regulate the expression of specific genes. The present study aimed to analyze the epigenetic profile of L1 between circulating tumor cells (CTCs) and non-cancerous samples. Methods: Blood samples were randomly collected from 18 patients, representing prostate, lung, breast, colon and pancreatic cancer and 5 healthy individuals. Cells were isolated by using enrichment protocols, including CD45 negative selection for normal samples and pancytokeratin positive selection for cancer samples. DNA and RNA were removed from the above cells and qPCR experiments were performed. The primers were designed to amplify specific regions of ORF1 and ORF2. The relative quantification was performed according to Livak method, by using 18SrRNA as housekeeping gene. All the reactions were performed in triplicates. Finally, oneway ANOVA was performed to compare the cancer and normal samples. Results: The analysis of DNA revealed the presence of both ORF1 and ORF2 in all samples. In the qPCR experiments, it is noteworthy that ORF1 RNA is not expressed in the majority of the samples in both groups. On the contrary, ORF2 RNA was expressed in all samples, however with different rates. The ORF2 was over-expressed in CTCs and the statistical analysis demonstrated a significant difference between CTCs and normal samples (p = 0.00089< 0.5). Conclusions: The L1 retrotransposon is found in all mammals and is the only active element in the human genome. According to literature and experimental data, the ORF2 is involved in stemness transcription factors. The present study demonstrated that ORF2 RNA expression is higher in CTCs than in normal cells, indicating once again the role of L1 in carcinogenesis. Further experiments, in more samples, should be performed to confirm the above and then to identify potential pattern of retrotransposition.

and glioblastoma cells as well as glioblastoma stem cells for anticancer treatment or radiation overexpressing opioid receptors. In this study, we analyzed different solid tumors in vitro, in mouse models and in cancer patients in vivo after D,L-methadone treatment in addition to conventional therapies. Methods: Different cancer cells of breast, ovary, pancrease, prostate, liver, lung, brain, colon, gastric cancer were treated with D,L-methadone alone or in combination with conventional therapies. Cell death and molecular cell death mechanisms were analyzed after D,L-methadone and/or anticancer drug treatment. Different cancer patients were treated with conventional therapies in addition to D,L-methadone. D,L-methadone was normally used in these patients for treatment of pain. In patients, tumor markers were measured and tumor growth was analyzed by computer tomography, magnetic resonance imaging, FDG-PET and FET-PET. Results: A strong induction of cell death was observed after combination therapy with D,L-methadone and anticancer drugs in vitro. Blocking of µ-opioid receptor signalling pathway inhibited the sensitizing effect of D,L-methadone to conventional therapies, inidicating that D,L-methadone sensitized tumor cells for anticancer treatment via triggering opioid receptor signalling pathways. In addition, strong inhibition of tumor growth, reduction of metastases, reduction of tumor size and reduction of tumor markers were observed after treatment with D,L-methadone in addition to conventional therapies in patients with breast, ovarian, pancreatic, lung, prostate, liver, colon and brain cancer. The tumors were progressive and/or refractory to conventional therapies. The tumor growth inhibition in patient were observed after substitution from morphine to D,L-methadone using for pain treatment . Conclusions: D,L-methadone triggering µ-opioid receptors seems to be a promising strategy for improvement of current conventional therapies in breast, ovarian, pancreatic, lung, prostate, liver, colon and brain cancer, resulting in a higher clinical outcome. Disclosure: No conflict of interest disclosed.

Posterdiskussion

Stammzellmobilisierung; Zellbiologie 2 P904

Cryopreservation of peripheral hematopoietic stem cells with uncontrolled freezing and 5% DMSO enhances CD34+ and CD45+ cell outcome compared to 10% DMSO Aurich K.1, Oergel T.1, Waterstradt M.2, Althaus K.1, Greinacher A.1, Krüger W.2 Universitätsmedizin Greifswald, Transfusionsmedizin, Greifswald, Germany, Universitätsmedizin Greifswald, Klinik für Innere Medizin C – Hämatologie und Onkologie, Greifswald, Germany 1 2

Introduction: A major limitation for successful treatment of cancer is development of resistance to conventional therapies. Therefore, novel treatment strategies are needed to improve therapeutic success. The therapeutic opioid D,L-methadone binds to opioid receptors. Cancer cells strongly overexpress opioid receptors. D,L-methadone sensitizes leukemias cells

Background: Controlled-rate freezing and storage in the vapour phase of liquid nitrogen is the standard technique for cryopreservation of peripheral hematopoietic progenitor cells (PHSCs). However, it is associated with high costs and dimethyl sulfoxide (DMSO) toxicity. Uncontrolled freezing, starting with 24 h at -80°C and subsequent transfer into the vapour phase of liquid nitrogen with only 5% DMSO instead of 10% DMSO preserves the functional capacities of PHSCs, and reduces toxicity during infusion. Especially in pediatric applications this is of importance, due to the lower bodyweight of the patients. In the present work, we have evaluated the impact of uncontrolled freezing and decreased DMSO concentration of 5% on CD34+ cell count, CD34+ and CD45+ cell viability and colony forming units (CFU)of ten autologous PHSCs compared to 10% DMSO. Methods: Autologous PHSCs from ten patients median age 59.0 ± 14.3 years, 8 male, 2 female) were produced by apheresis with Spectra Optia (Terumo, Germany) devices following volume reduction (320 g, 6 min) and addition of cryopreservation solution (autologous plasma/DMSO; 5 or 10% DMSO final concentration). Each PHSC was divided into 4 cryopreserved products, quick-freezed on dry ice, stored at -80 °C in a

Abstracts

Oncol Res Treat 2016;39(suppl 3):1–294

Disclosure: No conflict of interest disclosed. P903

D,L-Methadone increases the therapeutic success of conventional cancer therapies Friesen C.1,2, Schmidt R.1,2, Erhart D.1,2, Stalmach M.1,2, Alt A.2, Miltner E.1,2 Zentrum für Biomedizinische Forschung, Ulm, Germany, 2Institut für Rechtsmedizin, Ulm, Germany 1

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mechanical freezer for 24h before transferring into vp-LN2 for 28 days. Measurements of CD34+ and CD45+ cell counts and viabilities were performed with aliquots and determined by flow cytometry and the viability marker 7-aminoactinomycin D according to the ISHAGE protocol. Results: The impact of different DMSO concentrations on CFU, CD34+ and CD45+ cells during uncontrolled freezing procedure is shown in table 1. In summary, the use of only 5% DMSO was non-inferior to the standard using 10% DMSO. Tab. 1. Impact of different DMSO concentrations

5% DMSO

relative CD34+ cell count (% of 86.1 ± 14.3 initial cell count before freezing) CD34+ vitality (%) 99.1 ± 0.3 CD45+ vitality (%) 65.1 ± 9.7 CFU (×10 4/kg)

8.5 ± 4.5

10% DMSO p 67.6 ± 13.5

0.00005

97.9 ± 1.3 53.8 ± 14.9

n.s. 0.0006

9.4 ± 9.6

n.s.

Conclusion: Uncontrolled freezing in compliance with decreased DMSO concentration of 5% is practicable and results in enhanced CD34+ cell count, CD34+ and CD45+ cell viability as well as reduced DMSO related side effects. Disclosure: No conflict of interest disclosed. P905

Phenotypic assays using zebrafish haematopoiesis models for elucidation of hematopoietic toxicity Sauteur L.1, Lenard A.1, McGinnis C.2, Lenerke C.1,3 University Hospital Basel, Department of Biomedicine, Basel, Switzerland, Roche Pharma AG Innovation Center Basel, Basel, Switzerland, 3University Hospital Basel, Clinic for Hematology, Basel, Switzerland 1 2

Introduction: Hematologic findings are one of the most common side effects encountered in preclinical safety testing of new drug candidates. The consequences of direct or indirect damage to blood cells and their precursors can be potentially life-threatening, and hence, hematotoxicity can lead to the termination of a promising drug candidate. Current hematotoxicity testing employs in vitro models with a cell viability read-out. However, this approach only allows a limited read-out and, for example, does not capture effects on later maturation stages of blood cell progenitors. Methods: Here, we propose the zebrafish as an alternative animal model that captures the full range of hematopoietic lineages and maturation stages in an in vivo setting. Therefore, we use different transgenic zebrafish lines that specifically mark subsets of different hematopoietic lineages, flow cytometry, high content live-imaging and automated drug administration and image analysis. Results: We have recently established a drug-screening platform that assays the potential of chemical compounds on the erythroid lineage. We validated the antimitotic capacity of several cancer drugs, which prevented repopulation of mature erythrocytes after haemolytic anaemia. Now, we are applying this procedure to a large size compound library of FDA-approved chemicals. In addition, we are expanding the applicability of our technique to other zebrafish transgenic lines. In particular, we focus on macrophage populations and will test the effects of chemical compounds on this cell population after embryonic tail injury. Conclusions: We are now one step closer in introducing this novel multiplex in vivo model to predictive and mechanistic preclinical safety testing, which eventually will complement existing in vitro hematopoietic approaches. Disclosure: Loïc Sauteur: Employment or Leadership Position: Anstellung durch Roche; Expert Testimony: Ja Claudia Lenerke: No conflict of interest disclosed.

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P906

Successful peripheral blood stem cell mobilization using a cost-efficient single fixed-dose plerixafor schedule in poor mobilizers Greil C.1, Kiote-Schmidt C.1, Fink G.1, Hildenbeutel S.1, Bosse R.1, Duyster J.1, Engelhardt M.1, Wäsch R.1 Uniklinik Freiburg, Freiburg, Germany

Introduction: Collection of hematopoietic stem cells from peripheral blood (PB) is routinely conducted prior to high-dose chemotherapy and autologous transplantation. Despite safety and efficiency of current apheresis procedures there is still a significant rate of mobilization failures due to different patient-dependent factors necessitating additional agents like plerixafor. A standardized, cost-efficient strategy for its application in poor mobilizers is still lacking. Methods: As the number of patients with extensive pretreatment and long disease courses will even increase in the future, it is important to develop selection criteria for patients with expected benefit of additional plerixafor application, also with regard to cost efficiency. We analyzed 46 consecutive patients who received additional plerixafor for mobilization with the goal of a single fixed-dose at our academic center between 2011 and 2015. These patients were expected to be poor mobilizers due to low CD34+cell counts in PB prior to apheresis or after a first apheresis day with insufficient yield or after insufficient harvest with previous mobilizing chemotherapy (PB CD34+cells < 20/µl). We examined CD34+cell counts in PB and in apheresis products to identify patients who were able to collect a sufficient CD34+cell count for transplantation after a single application of plerixafor. Results: Plerixafor could be safely administered, leading in 83% and 48% to apheresis yields of >2 and >4×106 CD34+ cells/kg body weight (bw) and correlating with median PB CD34+cell counts of 8.0 and 17.8/ µl, respectively. 76% of patients showed a substantial benefit of plerixafor vs. colony-stimulating factor alone, with increased PB CD34+cells (13.3 vs. 4.7/µl, p = 0.0001) prior to apheresis and 4-fold higher CD34+cell numbers per single apheresis (1.2 vs. 0.3×106 CD34+/kg bw, respectively, p = 0.00005). A patient subset of 24% had < 5/µl PB CD34+cells before plerixafor application and profited less from additional administration. In 68% of patients plerixafor was administered only once, as planned. Conclusion: Our data suggest that most patients in preemptive and rescue settings with >5/µl PB CD34+cells can greatly benefit from a single fixed-dose application of plerixafor with median CD34+apheresis yield increases of 2.9 and median total apheresis collections of 4.6 x 106/kg bw whereas those with < 5/µl CD34+cells in PB may fail to substantially benefit and should therefore be carefully selected. Disclosure: No conflict of interest disclosed. P907

Benchmarking of the new Spectra Optia continuous apheresis system (cMNC) for peripheral blood stem cell collections Lisenko K.1, Pavel P.2, Schmitt A.1, Hundemer M.1, Shah S.1, WitzensHarig M.1, Ho A.D.1, Wuchter P.1 Universitätsklinikum Heidelberg, Abteilung Innere Medizin V, Heidelberg, Germany, 2IKTZ Heidelberg GmbH, Stem Cell Laboratory, Heidelberg, Germany 1

Introduction: The Spectra Optia system is a recently introduced apheresis system (Terumo BCT Garching, Germany) for mononuclear cell (MNC) collection equipped with a redesigned disposable kit and a new software program (version 11.2). The system enables a continuous collection of MNCs during the process, unlike the original Spectra Optia system (version 7.2), which included a chamber for two-step cell separation. The aim of this study was to comparatively analyze both apheresis systems according to specific performance parameters and the loss in platelet and hemoglobin during apheresis. Methods: A retrospective data analysis of 150 patients who underwent peripheral blood stem cell collection at our institution between March

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of 2014 and May of 2015 was performed. For the matched comparison, patients were grouped according to diagnosis and previous therapy: a homogeneous group of patients with multiple myeloma (MM) that had received first line therapy (“MM” group, n = 88) and a heterogeneous group of all other patients, including healthy allogeneic stem cell donors (“other” group, n = 62). To benchmark every LP session, we have compared the number of CD34+ cells collected with the number that was projected, and have assessed the performance ratio (collected/predicted CD34+ cells in %). Results: No significant differences in CD34+ collection yields between the two groups were found (pMM = 0.19, pother= 0.74). Moreover, similar performance ratios were observed (pMM = 0.89, pother = 0.1) and no relevant differences in platelet or hemoglobin loss were found between the two systems. Conclusion: We have demonstrated that for harvesting of CD34+ cells, the new version 11.2 Spectra Optia continuous collection system is as efficient as the original version 7.2 two-step separation system. No significant difference was found in terms of platelet or hemoglobin loss. We conclude that the new Spectra Optia cMNC system can be implemented with equal collection efficiency in a broad variety of patients intended for autologous transplantations, as well as in allogeneic stem cell donors. Disclosure: Katharina Lisenko: No conflict of interest disclosed. Patrick Wuchter: Advisory Role: Teilnahme an Advisory Boards von Sanofi-Aventis.; Financing of Scientific Research: Honorare von Sanofi-Aventis. Travel Grants von Hexal AG. P908

Platelet count before stem cell mobilization is associated with the need for plerixafor but not with CD34+ collection result Baertsch M.-A.1, Lisenko K.1, Pavel P.2, Bruckner T.3, Hundemer M.1, Ho A.D.1, Goldschmidt H.1,4, Wuchter P.1 Universitätsklinik Heidelberg, Hämatologie, Onkologie und Rheumatologie, Heidelberg, Germany, 2Institut für Klinische Transfusionsmedizin und Zelltherapie (IKTZ) Heidelberg, Stammzell-Labor, Heidelberg, Germany, 3 Universitätsklinik Heidelberg, Medizinische Biometrie, Heidelberg, Germany, 4 Universitätsklinik Heidelberg, Nationales Centrum für Tumorerkrankungen (NCT), Heidelberg, Germany 1

Introduction: Reports on peripheral blood stem cell (PBSC) mobilization have recurrently identified a low platelet count before mobilization as a significant risk factor for poor mobilization. To evaluate the relevance of this finding for PBSC mobilization with pre-emptive (PE) or rescue (R) plerixafor in case of poor mobilization, we performed a retrospective analysis of all patients undergoing PBSC collection at our institution between 01/2014 and 12/2015 (n = 380). Methods: Clinical and disease-related parameters, blood counts before mobilization, mobilization regimens, plerixafor administration and PBSC collection parameters were evaluated retrospectively. Data were evaluated for the overall cohort and for disease entities (multiple myeloma, lightchain amyloidosis, lymphoma, sarcoma and germ cell tumor), peripheral blood CD34+ cell count on day one of leukapheresis (≤/> 20 /µl) and need for PE/R plerixafor. A linear regression model was applied to analyze the relationship between the platelet count before mobilization and mobilization/collection results. Differences in blood counts among subgroups according to the need for PE/R plerixafor and mobilization success were analyzed by two-tailed Student´s t test. Results: 99% of patients (377/380) successfully collected ≥2×106 CD34+ cells/kg body weight sufficient for a single transplant. PE/R plerixafor was administered in 11% of patients (42/380). No correlation between the platelet count before mobilization and the CD34+ mobilization or collection result were detected, neither in the whole population nor in subgroups according to disease entities. However, patients requiring PE/R plerixafor had a significantly lower platelet count (217/nl vs. 245/nl; p = 0.004) and absolute neutrophil count (2.6/nl vs. 4.3/nl, p = 0.048) before mobilization. Patients with peripheral blood CD34+ cells on day one of leukapheresis ≤20/µl showed no significant difference in the platelet count before mobilization but had a significantly lower hemoglobin level before mobilization (10.9 g/dl vs. 11.6 g/dl, p = 0,004).

Abstracts

Conclusions: PBSC mobilization and collection was highly efficient using mobilization regimens incorporating PE/R plerixafor in case of poor mobilization. The platelet count before mobilization did not correlate with CD34+ mobilization and collection results but a low platelet count before mobilization was associated with the need for PE/R administration of plerixafor. MAB and KL contributed equally to this work. Disclosure: Marc-Andrea Baertsch: No conflict of interest disclosed. Patrick Wuchter: Advisory Role: Sanofi-Aventis; Financing of Scientific Research: Sanofi-Aventis; Other Financial Relationships: Hexal (travel grants) P909

Mobilization of autologous peripheral blood stem cells with the Spectra Optia® cell separator by continuous mononuclear cell collection protocol after cyclophosphamide and G-CSF in patients with multiple myeloma Klink A.1, Kunert C.1, Trinks U.1, Mügge L.-O.1, Sayer H.G.1,2, von LilienfeldToal M.1, Hochhaus A.1, Hilgendorf I.1 Universitätsklinikum Jena, Klinik für Innere Medizin II, Hämatologie und Onkologie, Jena, Germany, 2HELIOS Klinikum Erfurt, 4. Medizinische Klinik, Hämatologie, internistische Onkologie und Hämostaseologie, Erfurt, Germany 1

Introduction: Peripheral blood stem cells mobilized by chemotherapy with cyclophosphamide and G-CSF are often used in patients (pts) with multiple myeloma for autologous blood stem cell transplantation. The cell separator Cobe Spectra® MNC was recently replaced by the manufacturer with the succession type Spectra Optia®. The continuous flow process of MNC collection of this new cell separator offers the continuous mononuclear cell collection protocol (CMNC). Automated Interfaced Management system is designed to produce more consistent results through stability. However, the validation of the new production procedure for stem cell apheresis in the clinical setting is necessary. We compared the two cell separators by analyzing criteria of the stem cell product quality including the following cryopreservation procedures in two matched groups of pts. Patients and methods: In a validation study, 24 pts with multiple myeloma (median age 55 years, range 33-72, female n = 11, apheresis campaigns n = 43) were subjected to apheresis with Spectra Optia® to test feasibility and effectiveness from 12/2014 to 02/2016. Only stem cell mobilization regimens with high dose cyclophosphamide chemotherapy and application of stem cell growth factor G-CSF were selected and analyzed. Results were compared to a historical group with 24 pts (median age 59 years, range 48-70, female n = 11, apheresis campaigns n = 43) whose autologous apheresis had been performed with COBE Spectra MNC. Results: Most collection parameter, including collection efficiency and collection ratio, were similar. Platelet loss with Spectra Optia® CMNC (meanOptia14.9% ± 2.8) was significantly less than with COBE Spectra® MNC (meanSpectra 20.3% ± 2.7, p < 0.01). Products contained a lower hematocrit (meanOptia 3.2% ± 0.71 versus meanSpectra 4.1% ± 0.99, p < 0.001). No relevant differences were found with regard to quality features of cell products in both comparative groups, also including the viability of cells after cryopreservation, see table 1. Tab. 1.

Parameter CD34 x 10 total CD34 x 10 6 /kg body weight CFU-GM x 10 4/kg body weight Viability fresh (NC) trypan blue % Viability cryopres(NC) trypan blue % 6

Cobe Spectra (mean ± SEM) 344.8 ± 191.8

Spectra Optia (mean ± SEM) 328.5 ± 182.6

4.34 ± 2.61

4.58 ± 2.95

p = 0.69

64.6 ± 81.2

84.6 ± 62.7

p = 0.21

99.8 ± 0.28

99.9 ± 0.06

p = 0.19

97.6 ± 2.07

97.1 ± 1.83

p = 0.25

Oncol Res Treat 2016;39(suppl 3):1–294

p p = 0.69

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Conclusion: The procedure with the succession type of cell separator machine Spectra Optia® CMNC is estimated adequate and sure with similar efficiencies as COBE Spectra® MNC.

P911

A standardized in vitro blood-brain barrier model for studying nanocarrier transport at biological barriers

Disclosure: No conflict of interest disclosed.

Gräfe C.1, Slabu I.2, Müller R.3, Hochhaus A.1, von Eggeling F.3,4,5, Wiekhorst F.6, Clement J.H.1

P910

Comparison of two dose levels of cyclophosphamide for successful stem cell mobilization in myeloma patients Winkelmann N.1, Desole M.1,2, Hilgendorf I.1, Ernst T.1, Sayer H.G.1,3, Kunert C.1, Mügge L.-O.1, Hochhaus A.1, Scholl S.1 Klinik für Innere Medizin II, Universitätsklinikum Jena, Hämatologie/ internistische Onkologie, Jena, Germany, 2Helios Kliniken, Berlin, Germany, 3 HELIOS Klinikum, Erfurt, Germany 1

Introduction: Even in the era of bortezomib and lenalidomide, stem cell mobilization and harvest of CD34-positive cells for subsequent autologous stem cell transplantation continues to represent a standard approach for myeloma patients in first remission. Several mobilization regimens have been published while high-dose cyclophosphamide at a dose level of 4.0 g/m2 and subsequent stimulation with G-CSF has been intensively evaluated and is therefore commonly applied in patients with multiple myeloma. In contrast, intermediate-dose cyclophosphamide using 1.5 g/ m2 was not able to achieve a sufficient collection of stem cells. So far, the dosage of cyclophosphamide varies between different protocols affecting both the risk of chemotherapy-associated complications and the probability of successful stem cell harvest. Methods: We retrospectively analyzed the impact of 2.5 g/m2 (48 pts.) versus 4.0 g/m2 (53 pts.) cyclophosphamide in 101 consecutively evaluable myeloma patients (median age 59 years, range 32-72 years) who underwent stem cell mobilization in first remission. Successful mobilization chemotherapy was defined as a stem cell harvest of at least 5 million CD34 cells per kg bodyweight. Evaluation of toxicity especially considered infectious complications and hematological toxicity in both subgroups. Results: Successful stem cell mobilization was achieved in 40 of 48 (83%) and 44 of 53 (83%) patients, respectively. The median time to apheresis (11 vs. 12 days) and the median CD34 content of stem cell harvest (8.3 vs. 7.6 million CD34 cells per kg bodyweight) did not differ significantly between both groups. There was a significant difference of WBC nadir in favor of the cyclophosphamide regimen with 2.5 g/m2 (0.8 vs. 0.3 GPT/L, p = 0.021) and neutropenic fever was more often observed in patients who received 4 g/m2 cyclophosphamide (15% vs. 34%, p = 0.078). Importantly, after induction chemotherapy using the VCD regimen (bortezomib, cyclophosphamide, dexamethasone), successful stem cell mobilization was achieved in 26 of 29 (90%) patients treated with 2.5 g/m2 and 21 of 25 (84%) patients receiving 4 g/m2 cyclophosphamide, respectively. Conclusions: Cyclophosphamide at a dosage of 2.5 g/m2 is safe and highly effective for stem cell mobilization in patients with newly diagnosed myeloma in first remission after induction chemotherapy. Disclosure: Nils Winkelmann: No conflict of interest disclosed. Sebastian Scholl: Expert Testimony: Förderprogramm Infektiologie der Firma Gilead

Universitätsklinikum Jena, Hämatologie & Internistische Onkologie, Jena, Germany, 2RWTH Aachen Universität, AME – Institute of Applied Medical Engineering, Aachen, Germany, 3Leibniz-Institut für Photonische Technologien, Jena, Germany, 4Universitäsklinikum Jena, Klinik für Hals-, Nasen-, Ohrenheilkunde, Jena, Germany, 5Friedrich-Schiller-Universität Jena, Institut für Physikalische Chemie, Jena, Germany, 6Physikalisch-Technische Bundesanstalt, Berlin, Germany

Introduction: In general biological barriers such as the blood-brain barrier (BBB) constitute an obstacle for numerous drugs making sufficient and targeted drug delivery a mayor challenge. Thus, dedicated strategies and specially-designed drug delivery systems are developed. Based on their huge functionalization capacity superparamagnetic iron oxide nanoparticles (SPIONs) are highly attractive candidates for accomplishing not only drug delivery but also hyperthermal therapy and others. In order to study SPION passage through the BBB a well-defined test system is essential for generating comparative data. That is why this study aimed to establish a standardized in vitro test system to investigate the passage of SPIONs through the BBB. Methods: BBB-representing human brain microvascular endothelial cells (HBMEC) were used to establish cellular layers on porous transwell membranes. Barrier integrity was evaluated by transendothelial electrical resistance (TEER) measurements, molecular permeability assays, and immune-staining of tight junction protein ZO-1. Upon addition of differently coated SPIONs (140 nm) to tight cell layers SPION transport and permeation were assessed by confocal laser scanning microscopy, magnetic particle spectroscopy (MPS), and atomic absorption spectroscopy (AAS). Results: Barrier integrity studies reveal that HBMEC layer tightness can be modulated according to media supplements. Incubation of cell layers with cationic or anionic SPIONs compromised layer’s integrity in a concentration- and time-dependent manner indicating a layer-disrupting effect. In contrast, starch-coated SPIONs did not affect tightness parameters significantly. Additionally, SPION-specific iron quantification data showed that starch-SPIONs easily traverse cell-free membranes whereas the presence of cells compromised SPION crossing confirming high cell layer tightness. Moreover, both MPS and AAS indicate that starch-SPIONs are capable to overcome the BBB-representing layer in a time- and concentration-dependent manner without diminishing cell layer integrity. Conclusion: The presented workflow seems appropriate for further SPION transport- and nanoparticle-cell-interaction studies. Furthermore, the cell-mediated SPION transport and the SPION-mediated transport of immune cells across the BBB will be analysed. This work was supported by the DFG priority program 1681 (CL202/31, TR408/8-2, WI4230/1-2, MU2382/4-1) and Clinical Research Group KFO213 (TR408/4-3). Disclosure: No conflict of interest disclosed. P912

The siRNA – mediated MCM4 gene silencing for colorectal cancer treatment Kuzevanova A.1, Korotaeva A.1, Alimov A.1, Karpukhin A.1 Research Centre for Medical Genetics, Moscow, Russian Federation

Introduction: The RNAi based approaches for cancer treatment implies searching for new targets genes. The МСМ4 gene encoding the subunit of helicase, is involved into modulated tumor cell response to oxaliplatin. The purpose of this study to increase knowledge on MCM4 gene expression, evaluate the possibility of its use for new substance development. Materials and methods: The study conducted using colorectal cancer cells line HT-29.

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Real-time quantitative PCR was used to measuring gene expression. Lipid- based siRNA delivery was employed for gene silencing. Flow cytometry was applied for cell cycle analysis. Antibodies against AIF protein and fluorescent inhibitor of caspases-3 and -7 used for apoptosis study. Results: It was shown that low dose of oxaliplatin (1-5µM) up-regulates MCM4 gene mRNA expression more than 3-5 folds. The gene silencing induces apoptotic HT-29 cell death. The event is time- and dose-dependent. It was found that MCM4 silencing lead to accumulation of cells in G1 phase after 48 hours of exposure. Apoptotic cell death reached 60%. The analysis of apoptotic pathways revealed that apoptosis-induction factor is not involved in the activation of programing cell death. The main mechanism of cell death is associated with identified activation of caspases -3 and -7. Adding to the cell culture inhibitor of caspases-3 and -7 (Z-DEVD-FMK) significantly reduces the number of cells undergoing apoptosis. Conclusion: Thus, the activation of caspase-dependent pathway is the main cause of cell death after MCM4 gene silencing in presence of oxaliplatin. This opens up new opportunities to create target drugs, which should help to avoid the problem of resistance during oxaliplatin treatment. Disclosure: No conflict of interest disclosed. P913

Implementation of a Molecular Tumor Board in clinical decision making at the Medical Center University of Freiburg Claus R.1, Lutz L.2, Busch H.3,4, Csanadi A.2, Weddeling B.2, Fritsch R.1, Brummer T.3,4,5, Wehrle J.1, Miething C.1,4, Erbes T.6, Hettmer S.7, Brass V.8, Oehlke O.9, Demmer P.10, Meiss F.11, Peters C.3,4,5, Werner M.2,4, Börries M.3,4, Lassmann S.2,4, Duyster J.1,4, von Bubnoff N.1,4 Universitätsklinikum Freiburg, Klinik für Innere Medizin I, Hämatologie, Onkologie und Stammzelltransplantation, Freiburg, Germany, 2 Universitätsklinikum Freiburg, Institut für Klinische Pathologie, Freiburg, Germany, 3Institut für Molekulare Medizin und Zellforschung, Freiburg, Germany, 4DKTK, Standort Freiburg, Freiburg, Germany, 5BIOSS Centre for Biological Signalling Studies, Freiburg, Germany, 6Universitätsklinikum Freiburg, Klinik für Frauenheilkunde, Freiburg, Germany, 7Universitätsklinikum Freiburg, Zentrum für Kinder- und Jugendmedizin, Freiburg, Germany, 8 Universitätsklinikum Freiburg, Medizinische Klinik II, Gastroenterologie und Hepatologie, Freiburg, Germany, 9Universitätsklinikum Freiburg, Klinik für Strahlenheilkunde, Freiburg, Germany, 10Universitätsklinikum Freiburg, Institut für Humangenetik, Freiburg, Germany, 11Universitätsklinikum Freiburg, Klinik für Dermatologie und Venerologie, Freiburg, Germany 1

Introduction: In-depth knowledge about molecular pathogenesis of malignant diseases and rapidly increasing availability of targeted treatment options requires molecularly guided decision-making. We have established a Molecular Tumor Board (MTB) that focuses on patient management based on molecular data at the individual patient level. Methods: The biweekly MTB supports the work of organ-specific boards. Diagnostic and therapeutic recommendations are based on customized diagnostics and a case-by-case literature review. Diagnostic and therapeutic recommendations are communicated back to referring organ-specific boards. The MTB multidisciplinary team consists of expert physicians from Medical Oncology, Gynecological, Dermatological, Pediatric and Radiation Oncology as well as Pathology, Molecular biology, Computational Biology and Genetics. Results: The MTB was implemented in March 2015. Since then, a total of 100 pts have been discussed in 175 case discussions during 27 MTB meetings. Referred patient cases covered the entire range of malignancies seen by the organ-specific boards. In addition to routine diagnostics and panel sequencing with diagnostic reporting, customized diagnostics included exome, genome, transcriptome and methylome data. An in-detail analysis for 68 pts in 20 MTB meetings revealed a treatment recommendation rate of 43% (29/68 pts) and an implementation rate of 47% (15/32 recommendations). Treatment recommendations mainly comprised off-label antibody and tyrosine kinase inhibitor (TKI) therapy (41%) and trial inclusions (31%). Major reasons for non-adherence to recommendations

Abstracts

included patient will, death of pts and medical reasons. Based on MTB recommendations, off-label use of PD-(L)1 inhibiting antibodies was suggested in 11 cases. Five applications for insurance approval were filed and all were successfully approved. To date, in 5/15 (33%) pts with implemented treatment recommendations (TKI in- and off-label, antibody off-label), objective responses were observed, 2/15 (13%) pts achieved disease stabilization. Conclusion: Implementation of a MTB providing individualized treatment recommendations is feasible for a substantial proportion of patients in challenging clinical situations. The Freiburg MTB supports the work of organ-specific tumor boards. Particularly, rational off-label use of drugs and access to early clinical trials is enabled. Updated results will be presented at the meeting. Disclosure: Rainer Claus: No conflict of interest disclosed. Nikolas von Bubnnoff: Financing of Scientific Research: Fa. Novartis, Fa. BMS; Expert Testimony: Fa. Novartis P914

What does anakoinosis, communicative reprogramming of refractary tumors, accomplish? Reichle A.1, Ghibelli L.2, Gerner C.3, Herr W.4, Haegeman G.5 Universitätsklinikum Regensburg, Medizinische Klinik III, Regensburg, Germany, 2University Tor Vergata, Roma, Italy, 3Institut für analytische Chemie, Wien, Austria, 4Universitätsklinikum Regensburg, Regensburg, Germany, 5 Department of Physiology, Ghent University, Gent, Belgium 1

Introduction: The ancient Greek term “anakoinosis”, means “communication”. Communicative reprogramming, i.e. induction of anakoinosis in tumors aims at re-establishing novel communicative behavior of tumor tissues or between tumor tissue and hosting organism by re-modulating gene expression. Methods: Endogenous or therapeutically induced anakoinotic processes attribute novel validity and denotation to participators of biologic systems and facilitate in tumors by-passing so called undruggable oncogenic events with epigenetically (transcriptional modulators, epigenetic modifiers) and indirectly epigenetically modifying drugs (metronomic lowdose chemotherapy). The technical possibility and clinical tolerability of anakoinosis induction in refractory metastatic tumors could be shown in a large series of histologically quite different malignancies (Hart C et al. Cancer Microenvironment 2015). Results: In search for common denominators explaining results derived from intrinsically or extrinsically mediated anakoinosis, three main aspects could be separated, (1) the deduction of ubiquitously available communication rules (the metabolism of evolution, modular events, the tool of rationalizations for hallmarks in biologic systems, also in cancer, and the holistic communicative context), (2) the identification of cellular targets promoting communicative reprogramming (transcriptional modulators, epigenetically modifying drugs), and (3) among the effectors following anakoinosis induction with epigenetically modifying drugs, tumor suppressors, which dose-dependently contribute to anakoinotically constituting malignant behavior. Conclusions: The feasibility of anakoinosis induction, generally in biologic systems, demonstrates that ‘nature’, commonly investigated from a reductionist point of view, is now being reintegrated in a scientifically accessible social systems context: Targeted redemption of distinct and selectable communication processes proves to be the non-genetic therapeutic counterpart of therapies targeting oncogenic events. Pro-anakoinotic therapies integrate palliative care in the trajectory of cancer care. Disclosure: No conflict of interest disclosed.

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P915

Nicotinamide phosphoribosyltransferase (NAMPT) activates HCLS1 protein by deacetylation in a chronic phase CML (CPCML) Samareh B.1, Kuznetsova I.1, Loghmani-Khouzani H.1, Zikic A.1, Klimenkova O.1, Kanz L.1, Suttorp M.2, Welte K.1, Skokowa J.1 UniversitätsklinikumTübingen, Hämatologie, Onkologie und Immunologie, Tübingen, Germany, 2University Hospital Carl Gustav Carus, TU Dresden, Department of Pediatric Hematolgy and Oncology, Dresden, Germany 1

Introduction: Chronic myeloid leukemia (CML) is caused by BCR/ABL translocations and can be successfully treated with the BCR-ABL tyrosine kinase inhibitor (TKI) imatinib mesylate, or next-generations TKIs. While TKI- treatment inhibits the growth of CML cells, it does not eliminate them completely. In most patients, residual CML cells remain present and relapse is observed upon therapy discontinuation. CML patients must be continuously treated with TKI`s with risks of toxicity and a lack of compliance. CML cells may acquire mutations in BCR-ABL causing therapy resistance, blast crisis and mortality. Improved therapy for CML patients to make CML cells more susceptible to TKIs are necessary. Myeloid leukemia cells produce hematopoietic cytokines that induce proliferation and growth advantage. A better understanding of the pathways activated downstream of cytokine receptors in CML cells may help to efficiently target CML cells. We recently found that G-CSF activates protein deacetylation via Nicotinamide phosphoribosyltransferase (NAMPT). Furthermore, we described, that G-CSF phosphorylates and activates the hematopoietic-specific lyn-substrate 1 (HCLS1) protein. Methods: We investigated if HCLS1 is activated by the NAMPT-triggered deacetylation in CML. If this would be the case, targeting of NAMPT-triggered activation of HCLS1 may be a potential treatment of CML. Due to the specificity of HCLS1 protein for hematopoietic cells, HCLS1 inhibition will have no toxic effects in non-hematological tissues. We performed in vitro study of myeloid cell lines and primary CML cells in chronic phase (CP-CML) at diagnosis. Results: We found that NAMPT leads to deacetylation and activation of the hematopoietic-specific lyn-substrate 1 (HCLS1) protein. The specific NAMPT inhibitor FK866 markedly suppressed HCLS1 functions that led to the reduced proliferation and clonogenic capacity of CP-CML cells , unraveling a NAMPT/HCLS1 axis as a potential target to improve CPCML therapy. The effects of NAMPT inhibition on CP-CML cells were amplified by combination with Imatinib and had no toxic effects on hematopoietic cells of healthy individuals. Conclusions: We reported here a novel mechanism of the malignant evolution of CML cells via HCLS1 deacetylation by NAMPT. Our results raise opportunities to explore combination therapies of CML and other malignancies aimed at regulating protein functions by specific targeting of NAMPT-triggered protein deacetylation.

Posterdiskussion

Neue Substanzen, sonstige Onkologie P916

Patient-directed intervention to improve Quality of Life (QoL) for patients with soft tissue sarcoma (STS) undergoing palliative treatment: a multicenter, cluster-randomized, controlled trial of the German Interdisciplinary Sarcoma Group Hentschel L.1, Schuler U.2, Hornemann B.1, Schilling A.3, Freitag J.4, Richter S.4, Schuler M.K.4 Universitätsklinikum Carl Gustav Carus, Universitäts KrebsCentrum, Dresden, Germany, 2Universitätsklinikum Carl Gustav Carus, Universitäts PalliativCentrum, Dresden, Germany, 3Universitätsklinikum Carl Gustav Carus, Sozialdienst, Dresden, Germany, 4Universitätsklinikum Carl Gustav Carus, Medizinische Klinik I, Dresden, Germany 1

Introduction: In spite of evolving therapeutic options for the treatment of advanced STS, the balance between efficacy and toxicity still remains a major concern. Therefore, it is crucial to assess treatment effectiveness both as disease-related and Patient Reported Outcomes (PRO). Growing experience demonstrates the feasibility of electronic data capturing of PRO. Furthermore, the role of a structured multi-professional intervention for supportive care remains undefined in STS. YonLife trial (NCT02204111) is the first prospective trial assessing tailored palliative intervention in adult STS patients under palliative treatment with trabectedin. Methods: The YonLife trial is a multi-center, cluster-randomized, controlled, proof of concept study of patients with advanced STS to explore the effect of multi-professional expert-consensus derived treatment proposals on QoL. Adult patients undergoing treatment with trabectedin are being recruited in seven German hospitals. A 1:1 cluster randomization of the hospitals for multi-professional intervention or standard of care is applied. The seventh hospital serves as a reference center in order to pretest the planned study design. Patients are requested to answer standardized PRO-measures on a tablet-PC four times during nine weeks of treatment, which enables immediate scoring, adaptive testing and full Web access to the obtained data. The primary objective is the explorative analysis of the change of QoL-score after 9 weeks. Secondary objectives include feasibility aspects and changes of further PRO. Results: Enrollment began in October 2014 and so far 62 out of 77 planned patients have been included. Conclusion: In due course, the YonLife trial will provide more knowledge about QoL in patients with advanced STS. For the first time, PRO are electronically assessed and included in a prospective trial in this disease. Additionally, the benefit of tailored palliative intervention also will be investigated. Disclosure: No conflict of interest disclosed.

Disclosure: No conflict of interest disclosed. P917

Characteristics and treatment of sarcoma patients in a singlecenter analysis Glade J.1, Krogel C.1, Roessner A.2, Fischer T.1, Heidel F.3, Wolleschak D.1 Otto-von-Guericke University Magdeburg, Medical Center, Klinik für Hämatologie und Onkologie, Magdeburg, Germany, 2Otto-von-Guericke University Magdeburg, Medical Center, Institut für Pathology, Magdeburg, Germany, 3University Hospital Jena and Medical Faculty, Friedrich-SchillerUniversity (FSU) Jena, Internal Medicine II, Department of Hematology and Oncology, Jena, Germany 1

Introduction: Sarcomas contribute for 1% of all solid tumors in adults. The majority of sarcomas are soft tissue sarcomas. To assess for the patient characteristics and treatment modalities, we performed a retrospective single-center study in order to analyze all patients with a newly diagnosed sarcoma. Methods: In this single-center, retrospective study, we analyzed all patients with newly diagnosed sarcoma treated in our institution between

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November 1989 and March 2016. Primary objective of this retrospective study was to compare clinical characteristics and treatment modalities in different sarcoma subtypes. Results: Data from 139 patients were available for analysis. The main entities of all sarcomas in our analysis were the pleomorphic- (18%) and leiomyosarcomas (18%) followed by 10.8% lipo-, 7.2% chondro-, Ewingand synovial-sarcomas, 5.8% osteosarcomas and 11.6% others. The median age of the patient population was 50.9 years. At the time of primary diagnosis, 46% of patients were already metastasized. The initial therapy was surgery in 75.5% of all cases. 26.6% of all patients received adjuvant therapy. The combinations of ifosfamide (IFO) and doxorubicin (DOX) (37.8%) and combinations of vincristin, IFO, DOX, etoposid or actinomycin D and cyclophosphamid (43.2%) were the most frequently applied adjuvant therapies. IFO and DOX was also the most frequently used combination as first-line palliative therapy. Trabectidine was mainly used as second-line palliative therapy (36.4%). One of the determinants for adjuvant therapy and prognostic factors for survival is the tumor grading. Fifty-nine of our patients showed G3 at diagnosis. G2 was seen in 29.6% and G1 in 9.2% of all patients. The five-years overall survival of patients with G3 was 32%, G2 81% and G1 60%. Depending on tumor size and metastasis status we found median overall survival for T1 7.38 years, T2 3.06 years, M0 10.74 years and M1 2.19 years. The median overall survival in our whole patient population was 5.58 years. Conclusion: We report on a patient population of 139 individuals treated for sarcoma. The majority of patients underwent surgery as first line treatment. Ifosfamide and doxorubicin were the main pharmacologic agents used for adjuvant and palliative therapy. The median overall survival of all patients in our analysis was 5.58 years. As described in the literature patients with a differentiation grade 3 show, as well as in our study, a shorter five-year overall survival of 32%. Disclosure: No conflict of interest disclosed. P918

Improvement in ADCC and NK cell activation of an anticarcinoma bispecific antibody by genetic insertion of IL-15 as a cross-linker Schmohl J.U.1,2, Felices M.3, Miller J.S.4, Vallera D.A.5 Universität Tübingen, Hämatologie und Onkologie, Tübingen, Germany, University of Minnesota, Masonic Cancer Center, Section of Molecular Cancer Therapeutics, Minneapolis, United States, 3University of Minnesota, Department of Medicine, Division of Hematology, Oncology, and Transplantation, Minneapolis, United States, 4University of Minnesota, Department for Hematology and Oncology, Minneapolis, United States, 5University of Minnesota, Masonic Cancer Center, Section of Molecular Cancer Therapeutics, Therapeutic Radiology-Radiation Oncology, Minneapolis, United States 1 2

Introduction: Natural killer (NK)-cells are responsible for anti-tumor surveillance. Previously we constructed a bispecific NK-cell engager (BiKE) targeting CD16 (FcyRlll) on NK-cells and EpCAM on tumor cells (EpCAM16). Epithelial cell adhesion molecule (EpCAM) is a transmembrane protein expressed on a variety of carcinoma. The BiKE induced ADCC but didn´t induce expansion of effectors. To address this limitation, we incorporated a modified interleukin (IL)-15 crosslinker to create a trispecific NK engager (TriKE). This novel construction enhanced proliferation and activation of NK-cells. Methods: 1615EpCAM TriKE was assembled, expressed in E.coli, purified and refolded with a purity >90% and a molecular weight of 68,860 Da. Functional activity of the TriKE was assessed with 51Chromium release assays. Induction of proliferation, degranulation (CD107a expression) and Interferon (INF)-γ production was analyzed with flow-cytometry. Results: To assess the effect of IL-15 in drug activity, a 51chromium release assay with healthy PBMCs and EpCAM+ HT-29 colon carcinoma cell line in increased effector: target ratios (2.2:1, 6.6:1 and 20:1) was performed. Kill was increased with the TriKE compared to the BiKE and the effect was higher, the greater the presence of NK-cells. Furthermore PBMC were exposed to HT-29 and assessed for CD107a expression. NK-cells showed a significantly higher level with the TriKE compared to the BiKE or the

Abstracts

BiKE with additional IL-15 (p < 0.05). IL-15 alone did not enhance lytic degranulation or INF-γ, whereas exposure to the TriKE did. HL-60 acute promyelocytic leukemia cell line (EpCAM-) was treated simultaneously. No induction of degranulation and a minimum of INF-γ induction was visible. Specificity of stimulation was seen in induction of proliferation and prolonged survival. Using a reactive dye, expansion index was evaluated to be enhanced after exposure of purified NK cells to the TriKE and IL-15, whereas no enhanced proliferation was seen with the BiKE or the scFv monomers (p < 0.001). Conclusions: Whereas bispecific antibodies are known for their ability to enhance ADCC, the 1615EpCAM TriKE not only improves ADCC, but also enhances proliferation providing a selective self-sustaining signal to the NK-cell-target synapse. In summary, these data indicate that the TriKE platform represents a significant improvement over the bispecfic antibody platform in improving NK-cell related tumor defence. Disclosure: No conflict of interest disclosed. P919

Marine compound frondoside A exhibits high in vitro efficacy and toxicity in high risk neuroblastoma cell lines Otte K.1, Ackermann A.1, Kruchen A.1, Bleeke M.1, Dyshlovoy S.2,3, Müller I.1 University Medical Center Hamburg-Eppendorf, Department of Pediatric Hematology and Oncology, Hamburg, Germany, 2University Medical Center Hamburg-Eppendorf, Department of Oncology, Hematology and Bone Marrow Transplantation, Section Pneumology, Hubertus Wald-Tumorzentrum, Hamburg, Germany, 3G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Vladivostok, Russian Federation 1

Introduction: Despite recent therapeutic progress, high risk neuroblastoma patients still face a dismal prognosis due to the development of drug resistance. Induction of pro-survival autophagy by currently approved agents (doxorubicin, cisplatin) has been identified as a major resistance mechanism in high risk neuroblastoma. Thus, novel anticancer agents inhibiting pro-survival autophagy bear a great potential to improve overall survival in neuroblastoma patients. In this study, we examined the efficacy of frondoside A (FrA) - a marine triterpene glycoside – in a human neuroblastoma model. Methods: Anticancer activity of frondoside A was explored using the high risk neuroblastoma cell lines Kelly, IMR32 and LS. Effects on cell viability were investigated by MTT assay. The compound’s mode of action was analysed by light, fluorescent microscopy and immunoblotting. Additive effects of frondoside A in combination with cisplatin and doxorubicin were evaluated using the Chou-Talalay Method. Results: In vitro, FrA significantly reduced cell viability and inhibited proliferation of Kelly, IMR32 and LS in a dose-dependent manner with Kelly and IMR32 cells showing the highest sensitivity towards FrA. Notably, all tested neuroblastoma cell lines proved to be less sensitive to cisplatin (IC50s: 3-8 µM) compared to FrA (IC50s: 0.2-0.4 µM). FrA was found to upregulate LC3BII, a marker of type II cell-death and to induce autophagosome formation indicating that autophagy-associated processes are involved in the cellular response to FrA. Co-treatment with the established autophagy inhibitor bafilomycin A1 showed additive effects in all tested cell lines suggesting that FrA inhibits pro-survival autophagy. Moreover, FrA revealed additive effects in combination with standard chemotherapeutic drugs such as cisplatin and doxorubicin. Current research involves experiments to study FrA effect on pro-survival autophagy, HDAC10 expression, cell cycle progression and apoptosis. Conclusion: The marine compound frondoside A is a promising new therapeutic option for the treatment of high risk neuroblastoma displaying high in vitro efficacy and toxicity. The regulation of autophagy-associated processes is a supposed mechanism of action of the substance. Acknowledgement: This research is supported by Fördergemeinschaft Kinderkrebs- Zentrum Hamburg e. V. Disclosure: No conflict of interest disclosed.

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P920

Subgroup analysis of the phase III RADIANT-4 trial: Efficacy and safety of everolimus in advanced, progressive, nonfunctional Neuroendocrine Tumors (NET) of the lung Lahner H.1, Weber M.2, Hörsch D.3, Bojunga J.4, Vogel A.5, Voi M.6, Pacaud L.7, Pavel M.8 Universitätsklinikum Essen, Essen, Germany, 2Universitätsmedizin Mainz, Mainz, Germany, 3Zentralklinik Bad Berka, Bad Berka, Germany, 4Universitätsklinikum Frankfurt, Frankfurt, Germany, 5Medizinische Hochschule Hannover, Hannover, Germany, 6Novartis Pharmaceuticals Corporation, East Hanover, United States, 7 Novartis Pharma AG, Basel, Switzerland, 8Charité Universitätsmedizin Berlin, Berlin, Germany 1

Introduction: In the phase III RADIANT-4 trial, everolimus (EVE) improved median progression-free survival (PFS) by 7.1 months in patients (pts) with advanced, progressive, nonfunctional NET of lung or GI tract compared to placebo (PBO); HR, 0.48; 95%CI, 0.35-0.67; P < 0.00001. Aim of this subgroup analysis was to evaluate the efficacy and safety of EVE in pts with lung NET. Methods: The pts were randomized 2:1 to EVE 10 mg/d with best supportive care (BSC) or PBO with BSC. The present analysis shows the lung NET subgroup of the study population. Results: Of 302 pts, 90 had lung NET (EVE, n = 63 and PBO, n = 27). Median age was 65 years; 52% were males; 86% were Caucasian; most pts (99%) had well-differentiated disease; WHO PS 0/1/2 was 71%/28%/1%. Prior therapies (EVE vs PBO) included somatostatin analogues (mostly for tumor growth control; 43% vs 41%), surgery (52% vs 67%) and chemotherapy (40% vs 48%). Median PFS by central review (EVE vs PBO) was 9.2 (95% CI, 6.8-10.9) vs 3.6 (95% CI, 1.9-5.1) months with a risk-reduction in tumor progression by 50% in the EVE arm (HR, 0.50; 95% CI, 0.28-0.88). Most frequent (≥5%) G3/4 adverse events with drug-relationship (EVE vs PBO) were stomatitis (11% vs 0), hyperglycemia (10% vs 0), infections (8% vs. 0) and diarrhea (5% vs 0). Conclusions: Treatment with EVE improved the PFS by 5.6 months and reduced the risk in tumor progression by 50% in pts with advanced, progressive, nonfunctional lung NET compared to PBO. The EVE safety profile was similar to the overall RADIANT-4 population. Disclosure: Harald Lahner: Advisory Role: Novartis, Pfizer; Financing of Scientific Research: Ipsen, Novartis, Pfizer; Expert Testimony: Novartis; Other Financial Relationships: Ipsen, Novartis, Pfizer Marianne Pavel: Advisory Role: Ipsen, Lexicon, Novartis, Pfizer; Financing of Scientific Research: Ipsen, Lexicon, Novartis, Pfizer; Expert Testimony: Novartis, Ipsen P921

Secondary analysis from the phase 3 RADIANT-4 trial: Evaluating the impact of prior chemotherapy (CTx) on progression-free survival in patients with advanced, nonfunctional neuroendocrine tumors (NET) of lung or gastrointestinal (GI) origin Lahner H.1, Weber M.2, Hörsch D.3, Bojunga J.4, Vogel A.5, Voi M.6, Pacaud L.7, Pavel M.8 Universitätsklinikum Essen, Essen, Germany, 2Universitätsmedizin Mainz, Mainz, Germany, 3Zentralklinik Bad Berka, Bad Berka, Germany, 4Universitätsklinikum Frankfurt, Frankfurt, Germany, 5Medizinische Hochschule Hannover, Hannover, Germany, 6Novartis Pharmaceuticals Corporation, East Hanover, United States, 7 Novartis Pharma AG, Basel, Switzerland, 8Charité Universitätsmedizin Berlin, Berlin, Germany 1

Introduction: In the RADIANT-4 trial, everolimus (EVE) demonstrated an improvement in PFS by 7.1 months, compared to placebo (PBO; P < 0.00001) in patients with advanced, progressive, nonfunctional GI or lung NET. Methods: In RADIANT-4, patients were randomized (2:1) to EVE 10mg/d plus best supportive care (BSC) or PBO plus BSC. A subgroup analysis was performed to evaluate the impact of prior use of chemotherapy (CTx) on progression-free survival (PFS) in patients of the RADIANT-4 trial.

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Results: Of total 302 pts in the trial population, 77 (25%) had received CTx prior to study entry (EVE, n = 54 & PBO, n = 23) and 225 (75%) were CTx-naïve (EVE, n = 151 & PBO, n = 74). The baseline characteristics were comparable between subgroups. Primary tumor sites (prior CTx vs. CTx-naïve) were lung (49% vs. 23%), GI (38% vs. 65%) and NET of unknown primary (13% vs. 12%). The median PFS (95%CI) in the prior CTx group (EVE vs. PBO) was 9.2 (5.6-11.7) months vs. 2.1 (1.9-3.7) months (HR 0.35; 95%CI 0.19-0.64). In the CTx-naïve group (EVE vs. PBO), median PFS (95%CI) in the EVE arm was 11.2 (9.2-16.6) months vs. 5.4 (3.7-9.0) months (HR 0.60; 95%CI 0.42-0.86) in the PBO arm. Most frequent drug-related G3/4 AEs (EVE vs PBO) in the prior CTx group were stomatitis (11% vs 0), infections (9% vs 0), anemia (7% vs 4%) and diarrhea (6% vs 0); in the CTx-naïve group stomatitis (8% vs 0), diarrhea (8% vs 3%) and infections (6% vs 0). Conclusion: EVE improved the median PFS in patients with advanced, well-differentiated, progressive, nonfunctional NET of lung or GI origin, irrespective of the prior use of CTx. The safety profile of EVE was similar to the overall RADIANT-4 population. Disclosure: Harald Lahner: Advisory Role: Novartis, Pfizer; Financing of Scientific Research: Ipsen, Novartis, Pfizer; Expert Testimony: Novartis; Other Financial Relationships: Ipsen, Novartis, Pfizer Marianne Pavel: Advisory Role: Ipsen, Lexicon, Novartis, Pfizer; Financing of Scientific Research: Ipsen, Lexicon, Novartis, Pfizer; Expert Testimony: Novartis, Ipsen P922

Secondary analysis from the RADIANT-4 trial: Impact of prior Somatostatin Analogue (SSA) use on progression-free survival in patients with advanced nonfunctional Neuroendocrine Tumors (NET) of lung or Gastrointestinal (GI) origin Lahner H.1, Weber M.2, Hörsch D.3, Bojunga J.4, Vogel A.5, Voi M.6, Pacaud L.7, Pavel M.8 Universitätsklinikum Essen, Essen, Germany, 2Universitätsmedizin Mainz, Mainz, Germany, 3Zentralklinik Bad Berka, Bad Berka, Germany, 4Universitätsklinikum Frankfurt, Frankfurt, Germany, 5Medizinische Hochschule Hannover, Hannover, Germany, 6Novartis Pharmaceuticals Corporation, East Hanover, United States, 7 Novartis Pharma AG, Basel, Switzerland, 8Charité Universitätsmedizin Berlin, Berlin, Germany 1

Introduction: In the RADIANT-4 trial, everolimus (EVE) reduced the risk of disease progression or death by 52% compared to placebo (PBO; P < 0.00001) in patients with advanced, well-differentiated, progressive, nonfunctional Lung or GI NET. Aim of this secondary analysis in the RADIANT-4 trial was to evaluate the impact of prior use of SSA on PFS. Methods: Patients were randomized (2:1) to receive EVE 10 mg/day or PBO, both with best supportive care. This analysis reports baseline characteristics, PFS, and safety by prior SSA use. Results: Of 302 patients randomized, 163 (54%) had any prior SSA use (mostly for tumor control; EVE vs. PBO: 53% vs. 56%). Baseline characteristics were similar in patients with or without prior SSA. Primary tumor sites in the prior SSA group were GI (65%), Lung (23%), and NET of unknown primary (12%). Patients received ≥1 type of SSA, which included octreotide LAR (77%), octreotide SC (14%) and lanreotide (14%). The median duration of exposure to prior SSA was 15 months (range, < 0.1-103.5). The median PFS (central review; EVE vs. PBO) in the prior SSA group was 11.1 (95% CI, 9.2-13.3) months vs. 4.5 (3.6-7.9) months (HR 0.56; 95% CI, 0.37-0.85); in SSA naïve patients, 9.5 (8.2-16.7) months vs. 3.7 (2.4-8.1) months (HR 0.57; 95% CI, 0.36-0.89). The most common drug-related adverse events in the EVE arm (prior SSA vs. SSA naïve) were stomatitis (65% vs. 61%), diarrhea (34% vs. 28%) and infections (30% vs. 28%). Conclusion: Regardless of prior SSA use, EVE improved the median PFS in patients with advanced, progressive, nonfunctional Lung or GI NET. AEs were manageable and consistent with the overall study population.

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CONTENTS AUTHOR INDEX

Disclosure: Harald Lahner: Advisory Role: Novartis, Pfizer; Financing of Scientific Research: Ipsen, Novartis, Pfizer; Expert Testimony: Novartis; Other Financial Relationships: Ipsen, Novartis, Pfizer Marianne Pavel: Advisory Role: Ipsen, Lexicon, Novartis, Pfizer; Financing of Scientific Research: Ipsen, Lexicon, Novartis, Pfizer; Expert Testimony: Novartis, Ipsen P923

Somatostatin and CXCR4 chemokine receptor expression in gastroenteropancreatic neuroendocrine neoplasms (GEPNEN) of different origin and malignancy Mai R. , Kaemmerer D. , Sänger J. , Neubauer E. , Schulz S. , Lupp A. 1

Universitätsklinikum Jena, Institut für Pharmakologie und Toxikologie, Jena, Germany, 2Zentralklinik Bad Berka, Klinik für Allgemein- und Viszeralchirurgie, Bad Berka, Germany, 3Labor für Pathologie und Zytologie Bad Berka, Bad Berka, Germany 1

Introduction: Somatostatin receptors (SSTR), especially the SSTR2A, are known for their overexpression in well-differentiated gastroenteropancreatic neuroendocrine neoplasms (GEP-NEN). The chemokine receptor CXCR4, in contrast, is considered to be present mainly in highly proliferative and advanced tumors. Due to the scarcity of GEP-NEN, however, comprehensive data are still lacking on potential differences in the SSTR or CXCR4 expression pattern in dependence on the origin. Methods: Overall, 412 samples from 165 GEP-NEN patients, comprising both primary tumors (PT) and metastases (MTS), originating from the stomach, duodenum/jejunum, ileum, colon, rectum or pancreas were evaluated for the SSTR subtype 1, 2A, 3, 4, 5 and CXCR4 expression by means of immunohistochemistry using monoclonal rabbit anti-human SSTR or CXCR4 antibodies, respectively. The immunohistochemical stainings were evaluated by means of the Immunoreactive Score (IRS), comprising values from 0 to 12 points, and correlated to clinical data. Results: The SSTR2A was the most prominent receptor in the GEP-NEN samples investigated. It was present in 85% of the PT and in 77% of the MTS with a high intensity of expression, followed by the SSTR5 (PT: 23%, MTS: 21%), the SSTR3 (PT: 10%, MTS: 8%), the SSTR1 (PT: 9%, MTS: 3%) and the SSTR4 (PT: 4%, MTS: 4%). The CXCR4 was detected in 21% of the PT and in 14% of the MTS. With increasing malignancy of the tumors, significantly lower SSTR2A, but higher SSTR1, SSTR5 and CXCR4 levels were observed. PT displayed significantly higher SSTR2A expression levels as compared to the respective MTS. A low SSTR2A expression level was significantly associated with poor patient outcome. With respect to the different places of origin, both with primary tumors and with metastases, significantly lower SSTR2A, but slightly higher SSTR1, SSTR3, SSTR5 and CXCR4 expression levels were seen in (the generally higher malignant) tumors originating from the colon and the appendix as compared to those from other origin. Conclusions: As expected, the SSTR2A was the most prominent receptor expressed in the GEP-NEN samples investigated. However, expression levels varied in dependence on the extent of malignancy and on the location of the primary tumor. Especially in tumors originating from the appendix or colon quite low SSTR2A expression levels were observed. Here, pan-somatostatin analog or CXCR4 based diagnostics or treatment options should be taken into account. Disclosure: No conflict of interest disclosed.

P924

LOGIST ‐ A local observational GIST registry: A non‐ interventional registry to observe patients with gastrointestinal stromal tumors (GIST) after R0/R1 resection – with or without adjuvant therapy with imatinib (CSTI571BDE77) - an interim analysis with 5 years follow-up Reichardt P.1, Quietzsch D.2, Stübs P.3, Cameron S.4, Gellert K.5, Wilhelm M.6, Killing B.7, Croner R.8, Kopp H.-G.9, Will U.10, Wardelmann E.11, Krajinovic K.12 HELIOS Klinikum Berlin-Buch, Klinikum Buch Sarkomzentrum, Berlin, Germany, 2Klinikum Chemnitz gGmbH, Klinik f. Innere Medizin II, Chemnitz, Germany, 3Universitätsklinikum Magdeburg, Otto‐von-Guericke Universität, Klinik f. Allgemein-, Viszeral- u. Gefäßchirurgie, Magdeburg, Germany, 4 Universitätsmedizin Göttingen, Georg-August‐Universität, Zentrum Innere Medizin, Abt. Gastroenterologie u. Endokrinologie, Göttingen, Germany, 5 Sana‐Klinikum Lichtenberg, Oskar‐Ziethen-Krankenhaus, Klinik f. Allgem.- u. Viszeralchirurgie, Berlin, Germany, 6Klinikum Nürnberg Nord, Medizinische Klinik 5, Schwerpunkt Onkologie/Hämatologie, Nürnberg, Germany, 7LahnDill Kliniken GmbH Klinikum Wetzlar, Klinik f. Hämatologie, Onkologie u. Palliativmedizin, Wetzlar, Germany, 8Universitätsklinikum Erlangen, FriedrichAlexander Universität Erlangen-Nürnberg, Chirurgische Klinik, Erlangen, Germany, 9Universitätsklinikum Tübingen, Medizinische Klinik u. Poliklinik Innere Medizin II, Tübingen, Germany, 10SRH Wald‐Klinikum Gera, Klinik f. Innere Medizin III, Gera, Germany, 11Universitätsklinikum Münster, Gerhard-DomagkInstitut f. Pathologie, Münster, Germany, 12Universitätsklinikum Würzburg, Julius-Maximilians-Universität, Klinik f. Allgemein-, Viszeral-, Gefäß- u. Kinderchirurgie, Würzburg, Germany 1

Objective: The aim of the LOGIST registry is the long‐term evaluation of approaches in diagnosis, therapy and outcomes in patients with localized GIST and a different risk of relapse after surgical R0/R1 resection with or without adjuvant imatinib therapy. The maximum documentation period is 36 months or until recurrence or premature discontinuation, respectively. Results: 353 patients with localized GIST were documented in 66 participating sites since July 2010. The majority of patients were classified as high risk (27.1%), followed by low (23.4%) and very low risk (22.7%), predominantly by Miettinen method (51.1%). The mutation status was assessed in 64.6% of patients. Mutations of the KIT gene were detected in the majority of cases (69.3%), most often located in exon 11 (82.3%) while a PDGFR‐α mutation was found in 18.4% of patients, most often located in exon 18 (78.6%). Treatment with imatinib was administered in 46.7% of patients. In detail, 77 of 82 high-risk patients and 40 of 58 intermediate risk patients were treated with imatinib. Serious adverse events occurred in 24.2% of patients, however, the number of SAEs was reduced to 10.2% by case-bound consideration. Gastrointestinal disorders (grade 1/2) were the most often observed AE (28.3%). Causality between imatinib intake and a SAE could be established only in 6 cases from a total of 175 cases (7.4%), the number of SAEs was reduced to 3.0% by case-bound consideration. 23 patients (6.5%) developed disease recurrence. Follow up data for these patients such as mutation analysis, surgery, TKI treatment, time to progression, response rates, disease/survival status will be presented. Conclusion: LOGIST shows that patients with an intermediate or high risk of relapse according to Fletcher and/or Miettinen method are treated with imatinib in accordance with current ESMO guidelines. Data following disease recurrence have so far not been presented/published elsewhere. Imatinib treatment safety is comparable to non‐treatment safety. Real life efficacy data will be available in 2017. Disclosure: Peter Reichardt: Advisory Role: Pfizer, Bayer, PharmaMar, Ariad, Amgen, GlaxoSmithKline, AstraZeneca, Clinigen; Financing of Scientific Research: Novartis, Pfizer, Bayer, PharmaMar, Amgen, GlaxoSmithKline; Expert Testimony: Novartis Katica Krajinovic: No conflict of interest disclosed.

Abstracts

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P925

Initial non-functional metastases of a primary PTH-secreting parathyroid cancer – a case report Mayer K.1, Janzen V.1, Heine A.1, Brossart P.1 Medizinische Klinik III, Klinik für Hämatologie, Onkologie und Rheumatologie, Bonn, Germany 1

Introduction: Parathyroid carcinoma is a rare cause of hyperparathyreoidism. The clinical presentation is dominated by excessive sercretion of parathormone (PTH) with symptoms of hypercalcemia (bone disease and renal involvement) and less due to the infiltration of organs by tumor masses. Complete resection of the tumor is the treatment of choice. The follow-up is done with regularly monitoring of serum calcium and PTH levels thereafter. We here report a rare case of initial non-functional metastases after a primary PTH-secreting tumor. Case: The 63-year old male patient was diagnosed with parathyroid carcinoma and diffuse osteitis fibrosa cystica (“brown tumors”) in 2012. At the time of diagnosis, PTH was excessively elevated and hypercalcemia as well as renal insufficiency was apparent. Thyreoid lobectomy was performed and PTH decreased to normal levels afterwards. Of note, at that timepoint multiple nodular lesions were detected within the lung by CT-scan (not FDG-PET positive, in contrast to the primary lesion in the thyroid gland). Because of normalized PTH after resection, we decided to go for follow-up. During the follow-up lung nodules grew and new lesions in the lung appeared. However, the PTH level remained still normal. A biopsy from lung nodules was taken, which showed metastasis from the parathyroid cancer. Because of multiple lesions in the lung, surgical resection was not an option. Therapeutic options for metastatic disease are very limited and experience with chemotherapy is disappointing. Moreover, the patient refused any chemotherapeutic attempts. Two months later, the patient showed again signs of hypercalcemia and elevated calcium and PTH levels were confirmed. The hypercalcemia was treated with bisphosphonates. However, the metastases progressed during the next months also to the liver and the patient died 18 months after the verification of lung metastasis because of metastatic disease. Conclusion: This report shows the rare constellation of initial non-functional and then again functional metastases of a primary PTH-secreting parathyroid cancer. Therefore, this case emphasizes that surveillance of PTH levels alone are not reliable in the follow-up of parathyroid cancer and metastases cannot be ruled out through normal PTH-ranges. Routinely imaging and histological examination of suspicous lesions is recommended, even if PTH is not elevated and the tumor was former functional. Disclosure: No conflict of interest disclosed. P926

IDH1 mutation is associated with increased telomere length in glioma cells irrespective of WHO grade Ferreira M.V.1, Beier F.1, Bouillon A.-S.1, Sørensen M.D.2, Brümmendorf T.H.1, Beier D.3, Kristensen B.W.2,4, Beier C.P.3 Klinik für Hämatologie, Onkologie, Hämostaseologie und SZT, Med. Fakultät Uniklinik RWTH Aachen, Aachen, Germany, 2Department of Pathology, University of Southern Denmark, Odense, Denmark, 3Department of Neurology, University of Southern Denmark, Odense, Denmark, 4Institute of Clinical Research, University of Southern Denmark, Odense, Denmark 1

Introduction: Telomere length (TL) shortens with each cell division and reflects the replicative history of a cell. Cells with critical short TL can undergo chromosomal instability and thus promote cancer progression. Gliomas still have a dismal prognosis, especially after progression to highgrade gliomas. Little is known on how mutations of isocitrate dehydrogenase 1 (IDH1), the most important favorable prognostic factor, improve survival. Shortening of TL may explain the progression from low to highgrade gliomas, and we therefore studied TL in a cohort of glioma. Patients and methods: 68 gliomas WHO 2-4 (grade 2: n = 27, grade 3: n = 30, grade 4: n = 7) were included into this retrospective analysis. Clinical data, overall survival, age and IDH1 mutation status were available

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in all 68 patients. Median age of the patients was 30.1 years and 42% of the tumors had IDH1 mutations. Confocal Immuno-Q-FISH was used to analyze for TL in tumor tissue. Anti-alfa-SMA was used to selectively stain for endothelia cells. To reduce the large age and inter-individual variability, the relative TL (rTL) was determined on a single cell level by subtracting the mean TL of glioma cell TL from the mean TL of endothelial cells (internal control) in each tumor. IDH1 and proliferation of glioma was quantified based on routine immunohistochemistry. Results: rTL was significantly longer in IDH1 mutated tumors independent of the underlying WHO grade (grade 2: 18.8 arbitrary units (a.u.), grade 3: 26.2 a.u., grade 4: 20.1 a.u.) compared to tumors without IDH1 mutation (grade 2: 4.8 a.u., grade 3: 8.9 a.u., grade 4: 6.2 a.u., p < 0.001 for all tumors). Proliferation rate correlated with WHO grade, but was not significantly different (p = 0.06) between patients with and without IDH1 mutations. In contrast to IDH1 mutation and proliferation rate, TL was not an independent prognostic factor in a multi-variate analysis. Conclusion: We found that patients with mutant IDH1 have significantly longer telomeres independently of the histological grade possibly due to increased expression of telomerase preventing accelerated TL shortening. Telomere-mediated genetic stability in glioma might play an important role in patient with mutant IDH1. Further data is needed to clarify this issue. Disclosure: No conflict of interest disclosed. P927

CXCR4-directed PET imaging with [68Ga]Pentixafor in CNS malignancies Herhaus P.1, Habringer S.1,2, Gerngroß C.3, Slotta-Huspenina J.4, Tibor V.3, Wiestler B.5, Peschel C.1,2, Wester H.-J.2,6, Schwaiger M.2,3, Keller U.1,2 III. Medizinische Klinik (Hämatologie/Onkologie), Klinikum rechts der Isar, Technische Universität München, München, Germany, 2German Cancer Consortium (DKTK), partner site München and German Cancer Research Center (DKFZ), Heidelberg, Germany, 3Nuklearmedizinische Klinik, Klinikum rechts der Isar, Technische Universität München, München, Germany, 4Institut für Pathologie, Klinikum rechts der Isar, Technische Universität München, München, Germany, 5Neuroradiologie, Klinikum rechts der Isar, Technische Universität München, München, Germany, 6Pharmazeutische Radiochemie, Technische Universität München, München, Germany 1

Introduction: With the emerging role of targeted therapy in oncology, molecular in-vivo imaging of key targets provides a strong tool as biomarker for personalized therapeutic options and to refine response assessment. Proof-of-concept visualization of CXCR4 expression with the CXCR4-directed PET tracer [68Ga]Pentixafor has been shown in various malignancies. The chemokine receptor CXCR4 is a G-protein coupled receptor with CXCL12 as its sole known ligand. The CXCR4/CXCL12 system plays an important role during embryonic organogenesis and orchestrates important immunological functions and homing of hematopoietic stem cells to their niche. In lymphoma as well as in glioblastoma high CXCR4 expression is correlated with poor prognosis. As the CXCR4/CXCL12 axis promotes tumor growth, proliferation and survival by tumor-stroma interaction it might be a potential target for future targeted therapeutic strategies in those disease. Herein we describe the proof-of-concept of CXCR4-directed PET/MR imaging with [68Ga]Pentixafor in patients with CNS malignancies. Methods: 8 patients with CNS malignancies (3 glioblastoma, 2 DLBCL, 2 PCNSL and 1 extranodal MZL of the orbital cavities) were imaged with [68Ga]Pentixafor by PET/MR imaging after signing informed consent in this observational assessment. In one patient (extranodal MZL) response was assessed with [68Ga]Pentixafor by PET/MR imaging. Results: In 4 out of 8 patients – all with active disease – PET imaging with [68Ga]Pentixafor showed positive CXCR4 expression that correlated with the lesions determined by MR imaging. The standard uptake values (SUV) ranged from 3,6 to 23,0. One glioblastoma patient showed weak CXCR4 positivity (SUV 2,3) in a lesion in proximity to the resection side (positive in FET-PET). The CXCR4-directed response assessment in one

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patient with extranodal MZL showed reduced PET signal upon treatment (Figure1).

signed to one of the following treatment-groups: „go-go“, „slow-go“ or „no-go“. The results of the G8-screening were compared with results of the GA. Cut-off G8-values were ≥ 14 pts. for „go-go“-, 13-7 pts. for „slowgo“- and ≤ 6 pts. for „no go“-status. Results: The G8-screening as well as the advanced GA could be carried out easily within the daily clinical routine. Sensitivity of G8 was 0,74. Using only the G8-screening for therapeutic decision, 18% of all patients would have been undertreated while 8% would have been overtreated. G8-screening seemed to underestimate the patients capability to carry out a specific hemato-oncologic therapie compared to multi-dimensional GA. Conclusions: The G8-screening as well as the advanced GA are feasible in non-university hospitals. Performed by a well trained staff, they don’t require additional human or structural resources. G8-screening stays a useful screening tool for initial detection of geriatric impairment in elderly hamate-oncologic patients. Thus it lacks accuracy and can therefore not replace an advanced multi-dimensional geriatric assessment. Disclosure: No conflict of interest disclosed. P930

Geriatric assessment as treatment decision in elderly cancer patients Köster W.1, Willschrei H.-P.2, Stahl M.3 Gemeinschaftskrankenhaus, Herdecke, Germany, 2Malteser Krankenhaus St. Josefshospital, Krefeld, Germany, 3Kliniken Essen-Mitte, Essen, Germany 1

Fig. 1.

Conclusion: CXCR4-directed PET imaging with [68Ga]Pentixafor is feasible in patients with CNS malignancies. Due to the low uptake of this tracer in normal brain tissue and therefore good tumor to background ratio it might add valuable information during diagnostic procedure and response assessment. Disclosure: No conflict of interest disclosed. P929

Do G8-screening-results correlate with results of advanced geriatric-oncologic-assessments? von Saint-George T.1, Schaich M.1 RMK, Hämatologie, Onkologie und Palliativmedizin, Winnenden, Germany

Introduction: The aging society will lead to a significant increase in hemato-oncological diseases in the elderly for those individual diagnostic and therapy strategies are necessary. Appropriate geriatric diagnostics and individual therapy may therefore be carried out not only as part of studies or in specialized centers, but should be incorporated in daily clinical routine of every hemato-oncological department. For the detection of geriatric deficits some new screening tools as the G8-screening were lately developed. Our question is whether these short tests can replace a comprehensive geriatric assessment for therapeutic decision in the future. Methods: Since 11/2015 all our elderly patients (≥ 70 years) are screened on admission with the G8-screening tool for relevant geriatric impairments by trained nurses. Afterwards, for patients which were planned for a first-line therapy or a change in therapy (N = 50), a multi-dimensional geriatric assessment (GA) including ECOG-Status, Charlson-Comorbidity-Index (CCI), Activities of Daily Living (ADL), Instrumental Activities of Daily Living (IADL), Distress-Scale and Nutritional Risk Score (NRS) was performed by a physician. According to the results, patients were as-

Abstracts

Introduction: To assess the therapeutic options in untreated elderly patients (>= 70 years) with hematological or oncological diseases, we performed a geriatric assessment in all eligible patients over a period of 12 months. The aim of this data were to distinguish different patient groups who are able to be treated like younger patients (go-go´s), patients with limited treatment options (slow-go´s) and patients with only palliative care as best treatment (no-go´s) according to Balducci (Balducci and Extermann) Method: Following the advices of the “Geriatric working Group” of the AIO, the following tests were performed. - Barthel-Index (ADL) (nurse) - Instrumental Activity of daily Life (IADL) (occupational therapy) - Time-up & go-Test (physiotherapy) - Tinetti-Test (physiotherapy) - Mini-Mental-Status (psychotherapist) - Geriatric Depression Scale 15 (psychotherapist) - Body-Mass-Index (nutritionist) - Mini-Nutritional-Assessment (nutritionist) - Charlson-Score (physician) - Sozialfragebogen (family member) Afterwords, all patients were discussed in our muliprofessional, interdisciplinary tumor board. Results: 103 hematological/oncological elderly patients, 45 female, 58 male (gynecological patients were excluded), median age 82,2 years old. Barthel-Index 79 points (range 20 - 100) - Instrumental Activity of daily Life (IADL 6 points (range 1 - 8) - Time-up and go-Test 13 sec (range 5 - 35) - Tinetti-Test 19 points (range 2 - 28) - Mini-Mental-Status 25 points (range 8 - 30) - Geriatric Depression Scale 15 3 points (range 0 14) - Body-Mass-Index 26 kg/m² (range 16,7 - 43) - Mini-Nutritional-Assessment 18 points (range 3 - 26,5) - Charlson-Score 2 points (range 0 - 6) Conclusion: The presented data show clearly the wide range of different elderly patients between fit and unfit coming into a cancer center. An early executed geriatric assessment is able to realize the physical and mental capacity of elderly patients. Besides the clinical recognition of comorbiditys and multimedication and the causal malignancy, the social situation is necessary to realize the patients treatment options. The classification of Balducci is helpful in clinical practice. Disclosure: No conflict of interest disclosed.

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Posterdiskussion

Heranwachsende und junge Erwachsene (AYA), Langzeitüberlebende, Patientensicherheit P931

Adaption and validation of a questionnaire to assess the quality of life of young adult cancer patients with hematological malignancies Richter D.1, Mehnert A.1 Universitätsklinikum Leipzig AöR, Abteilung für Medizinische Psychologie & Medizinische Soziologie, Leipzig, Germany 1

Introduction: Cancer patients between 15 and 39 years, the so-called adolescents and young adults (AYA) have different needs in comparison to children and older adults during their cancer disease which reaches far beyond the aftercare and long-term survival. In Germany, only few studies examined the quality of life of AYAs due to the lack of age-appropriate instruments for measuring quality of life in this special age-group. Therefore, the aim of the present study was to translate and adapt a validated English-language questionnaire “Late adolescence and young adulthood survivorship-related quality of life measure” (LAYA-SRQL) into German and to evaluate the psychometric properties of the German version. This research was funded by the German José Carreras Leukemia Foundation. Methods: The original questionnaire consists of 30 items representing ten dimensions: spirituality, coping, fertility, sexuality, relationship, dependence, vitality, healthcare, education/career and cognition. The questionnaire was translated by two German researchers and native English speakers. We used forward and backward translation to preserve equivalence of the test questionnaire in the target language. The back-translated English version was checked regarding its congruence with the original LAYA-SRQL. In case of disagreements translations were discussed in the working group. Internal validity and reliability will be tested with factor analysis and Cronbach’s α. Moreover, we assessed convergent validity of the LAYA-SQRL subscales by examining correlations between SF-12 and PTGI. Reliability of the original English version of the LAYA-SQRL was good (>.70) and construct validity was strong. Results: The final version of the German LAYA-SQRL is currently being validated on a sample of n = 250 hematological survivors aged 15 and 39 years in an online survey. Participants will be recruited via social media, flyer in hospitals and cover letters. Recruitment will be completed in October this year. Conclusion: AYA-Survivors have a high need for support and information especially in the aftercare phase to get back to a normal life. With the adaptation and validation, a German questionnaire could be tested for the first time which is tailored to the specific needs of cancer survivors in the AYA-group. A screening-tool can greatly contribute to identify needs and deficits for improving psychooncological support and quality of life. Initial findings will be reported at the conference. Disclosure: Diana Richter: Expert Testimony: Studie wird gefördert im Rahmen eines Forschungsstipendiums der José Carreras-Leukämie Stiftung Anja Mehnert: No conflict of interest disclosed. P932

Novel approach for sociomedical counselling of young cancer patients Seifart U.1,2, Oldenburg M.1, Schulze J.1, Freund M.1 Deutsche Stiftung junge Erwachsene mit Krebs, Berlin, Germany, 2Arbeitskreis onkologische Rehabilitation in der DGHO, Berlin, Germany 1

Introduction: According to recent studies, about 80% of cancer patients between the ages of 18 and 39 survive their disease. However, in addition to the somatic and psychological side effects of the condition and its treatment, young patients suffer enormously from the short- and long-term social burden (financial and social consequences) of their disease and its treatment. Special aftercare is paramount. A return to the workplace is

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existential, especially for long-term survivors. The goal of this project is to advise patients on sociomedical (social law questions) early on to avert financial problems (possible threats) and prevent a decline in the quality of life. Methods: The YOUNG CANCER PORTAL of the German Foundation for Young Adults with Cancer is a nationwide, open-access, and free service for young patients. The advisory process rests on an independent database of sociomedical resources. Patients are initially asked to register with their personal information and their first questions. A questionnaire captures their current situation regarding the disease and daily life. Expert advice is given in writing, over the phone, or in person by a regionally assigned oncologist with extra training in social medicine. The database allows the anonymous collection and systematic analysis of the data. The goals are to analyze patients’ needs and problems, starting health policy debates, and improving the overall care. Results: The project started on 24 November 2015. So far 115 people have registered and 76 patients have been advised. Female patients make up 75% of the pool, male patients the other 25%. The average age is 31.9 years, and the age range is 20 to 53. The regional breakdown of the data shows that usage is nationwide. The questions focus on the patients’ professional situation and rehabilitation options, followed by questions concerning social security offices and insurance providers. A systematic analysis of support deficits and the development of concepts for improvement will follow. Discussion: The YOUNG CANCER PORTAL is unique throughout Germany and offers a new way for expert-patient-communication in the digital age. The portal’s basic structure enables the modular addition of topics. Throughout 2016, advice on endocrinological and cardiological questions will be added. A team focused on the topics of childbearing wishes and fertility will also be established. Disclosure: Ulf Seifart: Employment or Leadership Position: wissenschaftlicher Beirat der Stiftung junge Erwachsenen mit Krebs Mathias Freund: No conflict of interest disclosed. P933

Long-term toxicities and unmet needs of patients after cancer therapy – Analysis of the survivor register of the University Cancer Center Hamburg (UCCH) Quidde J.1, Fischer B.2, Koch B.1, Bokemeyer C.1, Haier J.1, Stein A.1 Universitätsklinikum Hamburg-Eppendorf, Universitäres Cancer Center Hamburg, Hamburg, Germany, 2Medical School of the University of Hamburg, Hamburg, Germany 1

Rationale: The number of cancer survivors is steadily increasing due to new therapeutic regimens. Within the next 20 years Germany will have to face estimated 4 million cancer survivors. Yet 89% of the survivors have unmet needs. This paper aims at assessing the present burdens and support requirements within the aftercare of cancer survivors – and focuses on AYAs (adolescents and young adults). Methods: Survivors participating in the survivorship program L.O.T.S.E. of the University Cancer Center Hamburg were enrolled. All patients answered standardized questionnaires about their strains of everyday life (Distress Thermometer), the requirement of support (SCNS SF), depression (PHQ-9), anxiety (GAD-7) and fatigue (FACIT-F version 4) as well as questions about demographic and financial facts. Survivors were differentiated in AYAs (18 to 39 years old) and adult survivors (older than 39 years). Results: 252 out of 259 survivors were enrolled, 59 AYAs and 192 adult survivors. Median age at diagnosis and survey inclusion was 49 and 55.5 years. The leading diagnosis were mamma carcinoma (21.9%), Non-Hodgkin lymphoma (5.5%) and sarcoma (8%). Hematological diseases (lymphoma, leukemia) were leading diagnoses for AYAs (58.3%) while solid tumors led amongst the adult survivors. General strains were high. AYAs reported about physical pain (42.4%), exhaustion (49.2%), eating/gastrointestinal problems (10.3-20.7%), polyneuropathy (18.6%), memory/concentration problems (37.3%) and emo-

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tional burdens like worrying (51.7%), fear (44.1%), nervousness (36.2%), sadness (22.4%), loneliness (15.5%), and sleeping problems (27.1%). Family development was being aggravated by sexual difficulties (11.9%), problems in dealing with the partner (6.9%). AYAs reported difficulties with finances (16.9%), troubles at school/work (21%). 5.2% AYAs had no work qualification and 22% were unemployed. 50% of AYA patients reported strains in the degree between “moderate” and “extreme”. Conclusion: AYAs have a high rate of unwanted side effects, strains and open needs – even 7.8 years (average, SD 8.7) after diagnosis. Individual health plus existential subjects like family and career development are deeply affected. Therefore specific and more comprehensive support within long-term follow-up programs in the clinical aftercare is essential. Disclosure: No conflict of interest disclosed. P934

The JETS-project of the German-Austrian-Swiss GVHD Consortium – a prospective study of late effects in adolescent and young adults after stem cell transplantation Hilgendorf I.1, Greinix H.2, Halter J.3, Ayuk F.4, Wagner E.-M.5, Lindner B.6, Gleich S.7, Hochhaus A.1, Wolff D.7, German-AustrianSwiss GVHD Consortium University Hospital Jena / Department of Internal Medicine II, Jena, Germany, Medical University of Graz, Division of Hematology, Graz, Austria, 3University Hospital Basel / Department of Hematology, Basel, Switzerland, 4University Cancer Center Hamburg-Eppendorf / Department of Hematopoietic Stem Cell Transplantation, Hamburg, Germany, 5University Medical Center of Johannes Gutenberg University Mainz / Department of Medicine III, Mainz, Germany, 6 Medical University of Innsbruck / Austrian Stem Cell Transplantation Registry (ASCTR), Innsbruck, Germany, 7University Hospital Regensburg / Dept. of Internal Medicine III, Regensburg, Germany 1 2

Introduction: Adolescent and young adults (AYA) present an understudied minority in the population of stem cell transplant recipients. AYA survivors are at high-risk to develop a wide array of late effects with impact on their quality of life. Material and methods: To capture the incidence and outcome of transplantation-associated late effects among AYA the German-Austrian-Swiss graft-versus-host disease (GVHD) Consortium launched a prospective study. Information about intensity of conditioning, occurrence of GVHD as well as the post-transplant multimorbidity index (PTMI) will be reported and stored into a web-based database on days + 100, +180, +360 after allogeneic stem cell transplantation (SCT). The database was programmed by the Evaluation Software Development (ESD) Company and is linked to the PROMISE-EBMT database sharing the MED-A data. Therefore, we can also capture non-adherent AYA, who withdraw from follow up visits at the participating transplant centers. In addition, a survey among all AYA reported to the registry will be performed at one year after SCT. Participants will receive a set of questionnaires covering the subject’s quality of life, psychosocial distress, personality, therapy adherence, physical activity, health-conscious behavior, vocational rehabilitation and consulting requirements. Results: We are still in the initiation phase of the project. All interested German speaking EBMT transplant centers are invited to participate, provided that they give their consent to the EBMT Registry office for data transfer to the registry. Conclusion: The JETS-project is based on the idea that the knowledge of sequel among AYA survivors of SCT including associated risk factors permits the development of a risk-adjusted follow up care plan for lifelong surveillance, implementation of preventive measures and counseling. Multidisciplinary and AYA-specific survivor concepts may help to improve knowledge, therapeutic adherence and empowerment of patients in order to further optimize medical treatment results and improve patients’ quality of life. Disclosure: Inken Hilgendorf: Expert Testimony: Hector-Stiftung Daniel Wolff: Expert Testimony: Hector-Stiftung, Carsten Bender-Leukämie Stiftung

Abstracts

P935

Long term observation after cure of cancer-patients beyond 10 years is necessary and meaningful. An experience of a monoinstitution between 1980-2015 Schroeder M.1, Wieschermann U.1, Aul C.1 HELIOS St. Johannes Klinik, Medizinische Klinik 2, Duisburg, Germany

Introduction: For a long time 5-year-follow-up was recommended as standard of care according to S3 guidelines for health insurance and sientific observation. But times have changed. People become older and life expectancy has extended. Time and content of follow-up recommendations are today under discussion. Cure rates have increased as a consequence of new therapy modalities. We see more often patients cured of their primary disease for a long time developing local relapse or second malignancies after more than 20 years. Therefore in contrast to the current modalities of follow-up outcome should be registrated for more than 5 years. Results: The follow-up´s of our investigated patients between 1980-2015 in our department will be presented. Methods: In our department we distinguish between 5 groups of risk-patients independently from their primary tumour stage and advise for these groups a longer follow-up. 1. Late relapses even after 20 or more years, e.g. breast-ca, melanoma, RCC. 2. Patients with cancer of the aero-digestive tract. 3. Drug-induced secondary malignancies (Etoposide, Anthracycline). 4. Patients suggestive for genetic predisposition. 5. Haematological malignancies after cured solid tumors. Conclusion: Due to a longer life expectancy risk of development of another malignancy has grown up. Registration of late side effects with long lasting lost of quality of life (physical & psychosocial health) has to be registrated. Disclosure: No conflict of interest disclosed. P936

O-PIS.meine akte – My patient record for my physicians Pareigis S.1, Kunde-Krüger J.2, Ebert A.2 SHG f. Leukämie- u. Lymphompatienten Halle (S.), Schkopau / OT Ermlitz, Germany, 2IT-Consult Halle GmbH, Halle, Germany 1

Question: Why do we need a web-based Patient Record for Patients? Background: A patient record can be comprised of many folders depending on the nature of the course of disease. The physical weight alone of carrying such folders, along with the tedious process of searching for relevant information is hardly acceptable. Solution: O-PIS.meine akte is a web-based application. It is accessible from any device (PC or Tablet) with Internet access. The patient digitizes and classifies his or her documents so that they can be centrally stored in O-PIS.meine akte. A physician will receive access credentials before the scheduled appointment with the patient. The physician can then securely access the patient’s record and add relevant documents as part of an ongoing treatment. Functionality: • Secure storage and classification of electronic documents • Efficient searching and information filtering • Chronological anamnesis • Apointment scheduling • Strict access and permission control • Medical documentation • Episode concept • Disease diary • Logging of treatments, diagnosis and findings • Emergency access Result: The O-PIS.meine akte web-based application makes it possible for patients as well as doctors to have fast and secure access to patient records from anywhere in the world.

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P938

Conclusion: The immediate interdisciplinary availability of reports on anamnesis, patient discharge, CT-MRT-PET data, course of disease, medication as well as ordered treatment is securely provided by means of using the O-PIS.meine akte application. Patients can independently upload documents such as referrals, vaccination cards, allergy IDs, availability and disease diaries. A pilot project is expected to start in the near future. Disclosure: No conflict of interest disclosed. P937

Systematic investigation of the medication process of cancer patients undergoing anti-neoplastic therapy Busse M.1,2, Krause S.W.2 Universitätsklinikum Erlangen, Apotheke, Erlangen, Germany, 2 Universitätsklinikum Erlangen, Hämatologie und internistische Onkologie (Medizin 5), Erlangen, Germany 1

Introduction: Owing to the complexity of anti-neoplastic therapy and the severe side effects of antineoplastic drugs, the improvement of medication safety in cancer patients has become an important topic in clinical routine and research. In the past, most research in this field focused on single putative sources of errors, mostly prescription as critical step. In contrast, our study provides a detailed analysis of the complete medication process of cancer patients undergoing intravenous anti-neoplastic therapy, i.e. recording of medical history, diagnosis, treatment planning, prescription, production, and administration. Methods: On a basis of 2×10 patients from the Department of Haematology and Oncology in Erlangen, checklists for structured documentation and evaluation were designed and validated in an iterative process. The tools thus developed were thereupon used to record the medication process of 100 patients by an “embedded researcher”. Results: The checklists developed in our study allowed the systematic evaluation of most of the critical steps along the medication process. In 100 therapy cycles we identified 171 deviations from an optimal process. Hindering factors turned out to be incomplete recording of medical history, insufficient consideration of other diseases (cardiac arrhythmias, respiratory diseases) and drug interactions leading to increased risk of long-QT-syndrome. Three more severe errors occurred: simultaneous application of Methotrexate and Cotrimoxazol, putatively resulting in delayed Methotrexate clearance, unintended prescription of Rituximab 375 (instead of 500 mg/m²) in CLL, and inappropriate interval between premedication and application of Rituximab leading to circulatory reaction. Seven additional errors were observed, but corrected before drug application: erroneous calculation of drug dosage, incomplete supportive therapy, wrong speed of infusion. Conclusion: The structured analysis of the total medication process by checklists proved useful to evaluate medication safety and identify possible errors. Critical steps like dose calculations should be subject to “double check” procedures. Besides these critical steps, “minor” deviations due to insufficient recording and consideration of the patients’ medical history may put patients at risk. It seems almost impossible to completely avoid such risks in the future without the help of a computerized physician order entry system tailored to needs of oncologists. Disclosure: No conflict of interest disclosed.

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Individual blister packaging and interdisciplinary patient care enables a cost-effective use of oral cytotoxic drugs Behrend M.1, Riederer C.2, Becker K.3, Wierecky J.4, Bertram M.5, Reschke D.6, Hegewisch-Becker S.7, Engel E.8 antares-apotheken oHG, Hauptapotheke, Hamburg, Germany, 2DAKGesundheit, Arzneimittel Steuerung, Hamburg, Germany, 3Onkologie Lerchenfeld, Hamburg, Germany, 4Schwerpunkt Onkologie Hämatologie Dres. Verpoort, Wierecky, Zeller, Hamburg, Germany, 5Hämatologisch Onkologischer Schwerpunkt Dres. Müller-Hagen, Bertram, Kollegen, Hamburg, Germany, 6 Onkologische Praxis Oldenburg/ Delmenhorst, Oldenburg, Germany, 7 Hämatologische-Onkologische Praxis Eppendorf, Hamburg, Germany, 8 Hämatologisch-Onkologische Praxis Altona (HOPA), Hamburg, Germany 1

Introduction: The packaging sizes of many high-priced oral cytotoxic therapies do often not consider the frequent necessity of dose modifications due to the side effects of these drugs and thus add to high costs because of discarded doses. Furthermore, tyrosine kinase inhibitors and other drugs often interact with concurrent medication influencing the pharmacokinetics, which might hamper efficacy or even increase side effects. Oral therapies represent a challenge which requires interdisciplinary patient care. The two main objectives of this project were to increase therapy safety and efficacy by establishing an efficient management of oral drug application and reduce costs of oral cytotoxic drugs by reducing the number of discarded doses. Methods: These aims were to be achieved by using individual blister packaging and especially attentive medical surveillance through an interdisciplinary approach of the treating oncologists and pharmacists. The oncologist had to define individual blister sizes for each patient. The following drugs were included in the project: capecitabine, erlotinib, everolimus, lapatinib, pazopanib, regorafenib, sorafenib and sunitinib. Between May 2013 until July 2015 45 patients were included. It was a collaborative project of the health insurance company DAK-Gesundheit, six private practices of oncology, the antares-pharmacy and the Onkotrakt AG. Medication related problems and the patients´ satisfaction were evaluated by self assessment questionnaires and regular telephone questioning. Results: Overall, costs with the different oral cytostatic drugs could be lowered by 10%. Drug interactions were identified in 34 of 45 patients. Of those, 16 were classified as relative contraindication, one would have lead to an increased and 13 to a decreased drug level of the applied oral medication. Patients’ satisfaction regarding the support of the team involved in the project and the individual blister packaging was very high. Conclusions: Optimization of oral cytostatic therapy can be achieved by monitoring drug interactions and applying intensive patient care realized by an interdisciplinary team of oncologists and pharmacists. In conclusion this project reveals that costs of oral therapy can be markedly reduced by using individual blister packaging. Interdisciplinary patient care was shown to be an effective way to significantly increase safety and quality of drug supply. The medical supply concept led to an overall high patient satisfaction. Disclosure: Milana Behrend: Employment or Leadership Position: Angestellte Apothekerin in der Antares-Apotheken oHG Erik Engel: No conflict of interest disclosed.

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P939

P940

Analysis and best solutions to prevent chemotherapy prescribing errors

Consultation hours for oral cancer therapy – a way to improve patient adherence and patient safety

Reinhardt H.1, Kaiser S.1,2, Otte P.1,2, Szymaniak-Vits M.1,2, Wöhrl S.3, Opeker K.1, Wolfrum P.1, Lieberwirth B.4, Ruch M.4, Duyster J.1,2, Jung M.5, Hug M.J.3, Engelhardt M.1

Vehling-Kaiser U.1, Drolle H.1, Damnali G.1, Kaiser F.2

University Medical Center Freiburg, Department of Hematology and Oncology, Freiburg, Germany, 2Comprehensive Cancer Center Freiburg, Freiburg, Germany, 3 University Medical Center Freiburg, Pharmacy, Freiburg, Germany, 4MPS GmbH, Freiburg, Germany, 5Albert-Ludwigs-Universität Freiburg, Institute of Pharmaceutical Sciences, Freiburg, Germany

Since the approval of the first tyrosine kinase inhibitor (TKI) called Imatinib in 2001 there has been a continuous increase of its usage in the therapy of malignant diseases. Today the oral anti-proliferative substances cover already 25% of total tumor therapy. In contrast, improving the patient adherence, thus ensuring the patient safety can´t keep up with the TKI-boom. However, since both treatment success and the occurrence of side effects directly depend on the patient adherence, it becomes more and more a focal point of interest. The WHO has come to the conclusion that the improvement of patient adherence is getting at least as much importance as the development of new TKIs. The control of oral therapy remains for the outpatient sector, in particular left to oncologists. In consideration patient education and monitoring time required are still missing remuneration nowadays; therefor new structures have to be created in oncological doctor’s offices. One possibility is the establishment of consultation hours specifically aiming oral tumor therapy which are being directed by a special trained medical assistant or nurse. Requirements for such consultation are a separate meeting room and sufficient time for an undisturbed patient consultation. During these special consultation hours, the patient is not only trained, but also medication intake and side effects are being monitored. About a year ago we introduced such a special consultation. At the beginning of oral therapy the patient is seen by a doctor and a special trained medical assistant together, then weekly inspection visits take place at the consultations, in addition every 4 weeks the doctor is being visited. The results obtained from the special trained medical assistant are discussed with inclusion of the doctor. This not only improves patient adherence but also saves time for doctors and patients. In a survey of 165 professional assistants for oral and subcutaneous tumor therapy showed a clear trend towards implementing their own counseling in everyday practice. Furthermore it is planned to investigate the necessity of a special consultation using a provision study.

Introduction: At Freiburg university medical center, chemotherapy (CTx) prescriptions are generated via an e-database (Chemo-AS) by the physicians and clinically checked by a surveillance team: the Clinical Cancer Research Group (CCRG) in close cooperation with pharmacy. Any detected error is instantly reported, corrected and recorded electronically. This system has now been established for over 10 years resulting in an interception of 99.9% of the CTx related errors. The main objective was to gain insight into causes and potential consequences of the CTx prescribing errors, serving as a foundation for the design of a next generation software with even improved safety features. Methods: A detailed analysis of 406 CTx prescribing errors prevented by CCRG initiative was performed using SPSS. The data had been collected prospectively over 2 years (2013/2014) via Chemo-AS by the CCRG. In an interdisciplinary approach, in cooperation with IT specialists, the causes of the errors were analysed for effective future preventability. Results: The 406 prescribing errors of 18.823 CTx orders were affecting 375 (2%) chemotherapy orders. Errors were divided into categories of potential consequences: overdose 203 (51%), underdose 59 (15%), incorrect length of cycle/CTx timing 56 (14%), wrong CTx 40 (11%), CTx not ordered 38 (10%) (with 10 errors additive in consequence). Of all errors analysed, 61% were identified as avoidable by specific software improvements. Results of the assessment of error preventability by program engineering are shown in Figure 1.

Tagesklinik für Hämatologie und Onkologie, Landshut, Germany, 2Universität Göttingen, Göttingen, Germany 1

Disclosure: Ursula Vehling-Kaiser: Advisory Role: Advisory Board; AbbVie, Roche Pharma AG, Lilly Deutschland GmbH, Amgen GmbH, GILEAD Sciences GmbH Florian Kaiser: No conflict of interest disclosed. P941

A phase 1 non-randomized open-label study to evaluate the effect of Regorafenib on probe substrates of CYP 2C9 (Warfarin), 2C19 (Omeprazole), and 3A4 (Midazolam) in a cocktail approach (Group A) and on a probe substrate of CYP 2C8 Rosiglitazone, (Group B) in patients with advanced solid tumors Gerisch M.1, Lettieri J.T.2, Boix O.3, Kelly A.2, Nieschwitz D.3, Sawyer M.B.4, Lin T.2, Diefenbach K.3, Haffner F.T.3, Radtke M.5 Bayer Pharma AG, BPH-DD-ED-DMPK-DMIC-DMW-DMW5, DM Wuppertal 5, Leverkusen, Germany, 2Bayer Pharamceuticals, Whippany, United States, 3Bayer Pharma AG, Berlin, Germany, 4Cross Cancer Institute, Edmonton, Canada, 5Bayer Pharma AG, Wuppertal, Germany 1

Fig. 1. CTx-error avoidability by program engineering.

Conclusion: This CTx prescribing analysis was very useful for pinpointing areas, where CTx error reduction by software engineering is possible resulting in a very safe therapy and also relieving doctors and pharmacy staff. The new upgraded CTx prescribing software tool is available to other external hospitals, leading to shared benefit for a large number of pts and healthcare staff. However as, according to our analysis, 30-40% of errors are not electronically avoidable, the CCRG, ward pharmacists or other surveillance teams remain indispensable. Disclosure: No conflict of interest disclosed.

Abstracts

Introduction: Regorafenib (REG) is an oral multikinase inhibitor approved for the treatment of mCRC and GIST. The primary objective of this study was to evaluate the effects of REG on pharmacokinetics (PK) of probe substrates of CYP 2C9 (warfarin), 2C19 (omeprazole), and 3A4 (midazolam) administered in a cocktail approach and on PK of a probe substrate of CYP 2C8 (rosiglitazone). Methods: This was an open-label, non-randomized, phase I study of REG 160 mg once daily in a 28 day (21 day on / 7 day off) cycle in 40 subjects with advanced solid tumors. The study was conducted at 4 sites. Subjects in Group A received a probe substrate cocktail containing warfarin (CYP

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2C9 substrate), omeprazole (CYP 2C19 substrate), and midazolam (CYP 3A4 substrate). Vitamin K (10 mg) was also administered to subjects in Group A to reverse the activity of warfarin. Subjects in Group B received rosiglitazone (CYP 2C8 substrate). On Day -7 of Cycle 1, a single dose of probe substrate(s) was administered without concomitant administration of REG. On Day 14 of Cycle 1, both REG and a single dose of probe substrate were administered. In Cycle 2 and subsequent cycles, only REG was administered. Blood samples for determining plasma levels of each probe substrate were collected up to 120 h post-dose; plasma samples for determining REG levels were collected up to 24 h after administration on Day 14 of Cycle 1. All samples were analyzed using validated LC-MS/MS methods. Results: Co-administration of REG with warfarin resulted in a mean increase of 25% in AUC, 19% in AUC(0-tlast) and 26% in the Cmax of S-warfarin compared with results obtained following administration of warfarin alone. There was a mean increase of 14% in AUC(0-tlast) and a mean increase of 16% in Cmax of 7-OH warfarin when warfarin was co-administered with REG compared with when given alone. The 6 hour post-dose omeprazole and 5-OH omeprazole data indicate a comparable metabolite-to-parent ratio, suggesting no inhibition of CYP 2C19. Co-administration of midazolam with REG resulted in a mean increase of 12% in AUC and a mean increase of 28% in Cmax of midazolam compared with midazolam alone. Co-administration of rosiglitazone with REG resulted in almost no change in the PK parameters for both rosiglitazone and its CYP 2C8 metabolite. Conclusion: Pharmacokinetic data indicate that REG may be given concomitantly with substrates of CYP 2C8, CYP 2C9, CYP 3A4, and CYP2 C19 without a clinically meaningful drug interaction. Disclosure: Michael Gerisch: Employment or Leadership Position: Bayer M. Radtke: Employment or Leadership Position: Bayer P942

Evaluation of CYP3A4 induction and inhibition on regorafenib and its major circulating metabolites in healthy subjects Gerisch M.1, Lettieri J.T.2, Boix O.3, Lin T.2, Diefenbach K.3, Hafner F.T.3, Radtke M.4 Bayer Pharma AG, BPH-DD-ED-DMPK-DMIC-DMW-DMW5, DM Wuppertal 5, Leverkusen, Germany, 2Bayer Pharamceuticals, Whippany, United States, 3Bayer Pharma AG, Berlin, Germany, 4Bayer Pharma AG, Wuppertal, Germany 1

Introduction: Regorafenib is an oral multikinase inhibitor approved for the treatment of colorectal cancer and gastrointestinal stromal tumors. The purpose of this study was to determine the effects of co-administration of a strong CYP3A4 inhibitor (ketoconazole) or inducer (rifampin) on the pharmacokinetics (PK) of regorafenib and its pharmacologically active metabolites M-2 and M-5. Methods: Two separate clinical studies with similar designs were conducted in healthy male volunteers. In Study 1, subjects received single doses of 80 mg (Cohort 1, n = 8) or 160 mg (Cohort 2, n = 18) regorafenib either alone or after 4 days of dosing with 400 mg ketoconazole once daily for 4 days, with ketoconazole dosing continuing for up to 7 (Cohort 1) or 18 (Cohort 2) days. PK of regorafenib, M-2, and M-5 were assessed after each dose. In Study 2, a total of 24 healthy volunteers received 160 mg regorafenib either alone or on day 6 of a 9-day regimen of 600 mg rifampin once daily. PK of regorafenib, M-2, and M-5 was assessed after each dose. Regorafenib and metabolites were assayed by LC-MS/MS. Results: Co-administration of ketoconazole with 160 mg regorafenib resulted in a mean 33% increase in regorafenib AUC and mean decreases in M-2 and M-5 AUC of 94% and 93%, respectively. Similar effects were seen on Cmax, with 40% increase in regorafenib, 97% decrease in M-2, and 94% decrease in M-5. Statistical results for Cohort 2 are summarized in Table 1. Results with Cohort 1 (80 mg regorafenib) were similar to those seen in Cohort 2. Co-administration of rifampin produced a mean 50% reduction in regorafenib AUC, a 363% increase in M-5 AUC, and no change in M-2 AUC. There was a 57% increase in mean M-2 Cmax with rifampin. Statistical results are summarized in Table 2.

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Tab. 1.

Tab. 2.

Conclusion: Co-administration of a strong CYP3A4 inhibitor increased the mean exposure of regorafenib and decreased the mean exposure of the pharmacologically active metabolites M-2 and M-5. Co-administration of a strong CYP3A4 inducer decreased the mean exposure of regorafenib, increased the mean exposure of M-5, and resulted in no change in the mean exposure of M-2. It is recommended to avoid concomitant use of strong inhibitors or inducers of CYP3A4 activity. Disclosure: Michael Gerisch: Employment or Leadership Position: Bayer M. Radtke: Employment or Leadership Position: Bayer P943

Advancing a chemotherapy (CTx) management software system to improve process & therapy results by key data analysis Szymaniak-Vits M.1, Klug J.2, Reinhardt H.1, Lieberwirth B.2, Kaiser S.1, Wolfrum P.1, Otte P.1, Haverkamp C.3, Schäfer T.4, Wuttke M.4, Hug M.5, Duyster J.1, Engelhardt M.1, Ruch M.2 Department of Hematology and Oncology, University Medical Center Freiburg, Freiburg, Germany, 2MPS – Medizinische Planungssysteme GmbH, Freiburg, Germany, 3Data Processing Center of the University Hospital of Freiburg, Freiburg, Germany, 4Meona GmbH, Freiburg, Germany, 5Pharmacy, University Medical Center Freiburg, Freiburg, Germany 1

Introduction: Since the complexity of CTx regimens increases, the risk of medication errors does likewise (Walsh, JCO 2008). In order to ensure patient (pt) safety, the Dept. of Hematology&Oncology, University Medical Center Freiburg approached this challenge by using a self-designed CTx-management software system (CTxMS). Combined with a comprehensive monitoring system and this CTxMS, we impressively prevent medication errors. We here describe the process of advancing this tool by implementing a central CTx-specific data base (DB) with a key data analysis tool to ensure and increase pt safety. Methods & Results: Our original CTxMS contains >500 detailed CTx protocols, including co-medication, SOPs and clinical pathways (Das Blaue Buch, Springer). Complementary to CTx corrections by the clinical pharmacy, additional monitoring – incorporated in a SAE management system – has been implemented within our CCCF. This is ensured by a quality management team, that allows to prevent almost all potentially harmful medication errors. Via software interfaces, our CTxMS is con-

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nected to the existing IT infrastructure to allow a seamless integration in up- & downstream processes. Based on this integration, the available therapy data is combined with that of the CTx processes (progress documentation, laboratory findings, study data, physicians´ letters, etc.). This hospital- & CTx-database can be analyzed regarding a wide range of relevant clinical questions, e.g. 1. the number of CTxs ordered and applied, 2. number of cancer pts being treated, 3. number of CTx-protocols performed within vs. outside clinical trials, 4. generation of key figures for certifications and recurring audits, and/or 5. data-mining analyses to optimize therapy patterns and CTx-processes. The flexibility of the software´s web-based architecture and interfaces make the simple transfer to other health facilities possible and supports physicians and another health care professionals to achieve similar outstanding results (Lohfert prize 2015). Conclusions: The UKF´s CTxMS ensures outstanding pt safety and efficient workflows. Extended to a central CTx-specific DB it’s possible to automatically generate key data, e.g. for CTx-controlling, process optimization and certification requirements. Our flexible technology allows a feasible transfer to other facilities, enabling other hospitals and private practices to achieve similar accomplishments despite restricted financial and technical resources. Disclosure: Magdalena Szymaniak-Vits: No conflict of interest disclosed. Markus Ruch: Stock Ownership: MPS – Medizinische Planungssysteme GmbH P944

Onkopti®- digitalized and standardized oncological therapy protocols in the internet Link H.1, Wickenkamp A.2, Huber J.1,3, Dresel A.1, Engert R.4, Kunz C.1, Lichtenberger S.1, Mahlmann S.1, Schäfer B.1, Zehnder-Kiworr C.1, Krämer I.5, Keßler C.6, van Lengen R.2, AIO German Cancer Society Westpfalz-Klinikum, Klinik für Innere Medizin I, Kaiserslautern, Germany, Fraunhofer Institute for Experimental Software Engineering IESE, Process Engineering, Kaiserslautern, Germany, 3Schwerpunktpraxis für Hämatologie und Onkologie, Bad Kreuznach, Germany, 4Westpfalz-Klinikum, Klinik für Innere Medizin II, Kaiserslautern, Germany, 5Universitätsmedizin der Johannes Gutenberg-Universität, Apotheke, Mainz, Germany, 6Westpfalz-Klinikum, Zentralapotheke, Kaiserslautern, Germany 1 2

Oncological protocols for pharmacotherapy of cancer are compiled from original papers, guidelines, textbooks, collections of protocols, study groups and congress articles. Creating and updating completely new protocols is complex, time consuming and unstandardized. For these reasons, implementation of permanent and numerous innovations in clinical standards is often considerably delayed. Furthermore, a central quality assurance is neither in place nor is there a functioning variant management of routine clinical protocols. The information system Onkopti® (www.onkopti.de) provides clinical oncologists with current and editorially screened data on routine oncological protocols and study protocols for solid tumours and haemoblastoses via download from the internet. In addition, all protocol-relevant information, such as literature citations, medical societies’ recommendations and guidelines, toxicity as well as information on supportive and adjuvant therapy, advice for carrying out the protocols, control parameters, etc. are captured and can be pulled up, as needed. Working together with external experts and study groups, which contribute to the editorial work flow, experienced specialists in oncology, haematology and pharmacy review the internet protocol databank in form and content and maintain constant quality assurance. After successful validation, the protocol is released. A protocol can be viewed and printed out in different degrees of scope and detail. Over 700 protocols are available and are amended continuously. A mini-version is freely available in the internet. The attending physician can simply download the standardised treatment protocol in the form of an Excel document. This document also includes supportive and adjuvant therapy and allows the physician to generate a patient-specific, daily medication schedule. Exporting complete protocols in the Cato programme for individualized therapy, adjuvant care and preparation of cytostatics in the pharmacy is part of daily routine practice.

Abstracts

Onkopti® offers digitalised, standardised therapy protocols for download and enables further use of complete protocols in computerized physician order entry systems. The resources for protocol generation can be centralised, thereby freeing up the user’s resources. The central internet-based databank Onkopti® enables pharmacists and physicians to establish and conduct optimal oncological therapy which is state of the art in terms of quality assurance and standards. Disclosure: Hartmut Link: Honoraria: Onkopti: Urheberrecht Rolf van Lengen: No conflict of interest disclosed. P945

Combination therapies in oncology – the developments regarding added benefit and costs during the first 5 years of early benefit assessment Schubert A.1, Tebinka-Olbrich A.1, Zentner A.1, Haas A.1 GKV-Spitzenverband, Arznei- und Heilmittel, Berlin, Germany

Introduction: Combination therapy as a therapeutic intervention is common in oncology. Several treatment regimens that include two or more therapeutic agents have been authorized over the past years and been subject to the early benefit assessment of pharmaceuticals by the Federal Joint Committee (G-BA) in Germany since 2011 in accordance with the Act on the Reform of the Market for Medicinal Products (AMNOG). Methods: The G-BA’s resolutions regarding the early benefit assessment of pharmaceuticals are summarized in terms of the proportion of combination therapies assessed in oncology and per oncological indication. Furthermore, the appropriate comparators as well as the additional benefit in relation to the appropriate comparators are summarized and grouped per year, orphan drug status and therapeutic indication and contrasted with the additional benefit claimed by the pharmaceutical companies in their benefit dossiers. With regard to patient relevant endpoints, results overall survival, morbidity, health-related quality of life and adverse events are presented for each combination therapy. Finally, the costs of combination therapies and their appropriate comparators according to the FJC are presented. Results: The proportion of combination therapies which are subject to the AMNOG-process of early benefit assessment is rising. Often, new active ingredients are combined with therapies that have gained marketing authorization several years ago and are compared with therapies for which the benefit was never assessed. Costs per treatment are steadily increasing, posing a significant challenge for the statutory health insurances. Conclusion: In addition to combinations of newly authorized agents many older therapeutic agents whose additional benefit has never been assessed are also combined, or new and old agents together constitute a treatment pattern. Combination therapies therefore have always been an important aspect of oncological treatment, but this trend is continued by the latest marketing authorization. However, the sometimes considerable uncertainty of the underlying clinical data especially with regard to overall survival as well as the escalating treatment costs give reasons for concern. Disclosure: No conflict of interest disclosed.

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Posterdiskussion

Supportivtherapie, Infektionen P946

Pilocarpine for prophylactic and therapeutic treatment of radiation induced xerostomia in patients with head and neck cancer Werner J.1, Jahn F.1, Riesenbeck D.2, Unverzagt S.3, Müller-Tidow C.1, Jordan K.1 Universitätsklinikum der Martin-Luther-Universität Halle-Wittenberg, Klinik für Innere Medizin IV, Hämatologie/Onkologie, Halle (Saale), Germany, 2 Strahlentherapeutische Gemeinschaftspraxis, Recklinghausen, Germany, 3 Institut für Medizinische Epidemiologie, Biometrie und Informatik, Halle (Saale), Germany 1

Introduction: Xerostomia is a very common side effect of radiation in patients with head and neck cancer, occurring to some degree in up to 100% of patients. Saliva gland injury reduces saliva production, so patients have a dry mouth and report difficulty with chewing and swallowing food. The question of the systematic review was whether Pilocarpine is beneficial as a prophylactic or therapeutic agent in patients with radiation induced xerostomia. Methods: Literature search was conducted from 1993 until 2015 using database CENTRAL and Medline (Pubmed, Ovid). Search was limited to trials with adult patients, published in English or German. Only randomized controlled trials (RCTs) and meta-analyses were included. Primary endpoint was severity of radiation induced xerostomia (CTCAE grade 1-4). Results: In the systematic review we included 49 trials for all prophylactic and therapeutic treatments of radiation induced xerostomia. Six trials referred to prophylactic use of pilocarpine (n = 729). These compared pilocarpine with different doses or placebo. In regard to high radiation doses (>50 Gy) pilocarpine turned out to be effective in increasing unstimulated whole salivary flowrates versus stimulated saliva flowrates at end of radiotherapy (28% vs. 15%, p = 0.05). Pilocarpine did not lead to a significant difference in RTOG-xerostomia severity as assessed by LASA (MD: 1.30; 95% KI:-9.43-12.03). There was no significant difference between pilocarpine and placebo for saliva production or xerostomia (MD: 13.30; 95% KI: -1.65-28.25). Patients reported less oral discomfort with pilocarpine versus placebo during radiotherapy (p = 0.001). Five studies on therapeutic use of pilocarpine were available (n = 754). Results indicate that oral pilocarpine after radiotherapy is more effective than placebo. Patients given pilocarpine demonstrated significant improvement of dry mouth symptoms versus placebo (RR: 2.00; 95% KI: 1.21-3.29; RR: 1.77; 95% KI: 1.15-2.70). Studies reported mild adverse events in patient with higher pilocarpine doses (10 mg) after radiotherapy. Conclusion: This systematic review indicates that prophylaxis with pilocarpine during radiotherapy can improve unstimulated whole salivary flow rates, but has no influence on symptom-reduction of xerostomia. Therapeutic use is effective in terms of reducing symptoms in patients with radiation induced xerostomia. These results formed the basis of specific recommendation within the S3 guidelines “Supportive therapy”. Disclosure: No conflict of interest disclosed. P947

The influence of a qualified nutrition counseling and intervention upon the progress and quality of life of patients with lung cancer Kiesel M.1 Julius-Maximilians-Universität Würzburg / Missionsärztliche Klinik Gemeinnützige Gesellschaft mbH Würzburg, Würzburg, Germany 1

The problem of malnutrition is proven for many tumor entities. For lung cancer patients there is no clear information for the probable effect of measurement and intervention regarding nutrition.

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In this study, 83 patients with lung carcinoma were included and divided in two groups. All were informed about the issue of nutrition of cancer patients. Data during therapy was collected for 6 months, including Body Mass Index (BMI) and lean mass (fat free mass, called Phasenwinkel alpha). The latter was measured via Bioelectrical Impedance-Analysis (BIA). Group 2 was asked additionally three times about their quality of life as well as nutritional behavior via questionnaires at month 0, 3, and 6. These consisted of the international EORTC QLQ-C30 questionnaire as well as questions about the patients’ nutrition. Most of the questions were to be answered by choosing on a scale of one to four points. Both BMI and lean mass were generally deteriorating during the 6 months by 0,79 kg/m2 and 0,96 ° in group 1 and 0,216 kg/m2 and 0,56 ° in group 2. The correlation coefficient (cc) of the progress of BMI and lean mass showed 0,07 in group 1 and -0,15 in group 2. In group 1 the maximum loss of BMI and lean mass was in patients who had undergone radiation. In group 2 the maximum loss was in patients who had undergone operation. Patients with multimodal therapy did not loose more BMI or lean mass than those with unimodal therapy. The cc of BMI and lean mass was up to 0.97 (p = 0,01) concerning low intake of dairy products, lack of concentration, personal judgment of own life quality and health, social impairment, fatigue, dysphagia and xerostomia, etc. The questionnaires showed that generally life quality and nutritional behavior decreased. The average difference of the points given varied around 10%. Few vegetables and fruit in the diet lead to more impairments in daily life and a lack of concentration (cc = 0,65; p = 0,01). Similar findings could be made for little meat- and fish-uptake. Uni- and multimodal therapies affect patients’ nutrition equally. There was no correlation between the progress of BMI and lean mass. During lung cancer treatment, life quality decreases in multiple ways. This correlates with a general decrease in nutrition and in healthy nutrition in particular, emphasizing the need for nutritional counseling and diagnostics. It seems to be of no additional use to measure lean mass in addition to BMI. Further research is recommended on this issue. Disclosure: No conflict of interest disclosed. P948

Implementation of G-CSF-guidelines to prevent febrile neutropenia after chemotherapy in patients with lung or breast cancer – second representative sample survey in Germany Link H.1, Holtmann L.2, Kerkmann M.2, Ortner P.3, ASORS and AIO German Cancer Society Westpfalz-Klinikum, Klinik für Innere Medizin I, Kaiserslautern, Germany, 2MMF GmbH, Dortmund, Germany, 3POMME-med GmbH, Munich, Germany 1

Prophylaxis of febrile neutropenia (FN) with G-CSF after chemotherapy (CTX) is recommended in guidelines (GL), if the risk FN is high (≥20%), or intermediate (≥ 10% - 20%) in case of additional risk factors. A sample survey in 2012 (NP1) showed a lack of GL-adherence (GLAD) (Link H, Support Care Cancer 2016;24:367). This second survey evaluated if GLAD and GL implementation have improved. The sample size represented 1.0% of the incidence of lung and 1.1% of breast cancer in Germany in 2010. Data of pts treated with at least 2 cycles of CTX with a FN risk ≥10% between 10/2014 to 9/2015 were documented retrospectively. Data from 573 lung cancer (LC) and 801 breast cancer (BC) pts was collected from 109 hospitals and 83 oncology practices with 222 physicians participating. Compared with the NP1 survey, GLAD increased in LC and FN-high risk (HR) CTX from 15.4% to 47.8% (p < 0.001), and in FN-intermediate risk (IR) CTX from 38.8% to 44.3% (p = 0.003). In BC and FNHR CTX, GLAD was unchanged: 85.6% vs. 85.1% (p = 0.73), but increased in FN-IR from 49.3% to 57.8% (p < 0.001). G-CSF was used in 74.2% of all 1st FN-HR CTX cycles and in 77.5% of all subsequent cycles (p = 0.19). In all FN-IR CTX cycles, G-CSF prophylaxis was used significantly less in the

Abstracts

CONTENTS AUTHOR INDEX

first CTX-cycles (27.0%) than in the subsequent cycles (37.2%, p < 0.001). In pts treated in certified or comprehensive cancer centers, the GLAD in FN-HR CTX was 80.3% vs 72.4% in other centers (p = 0.001), in FN-IR CTX it was 53.8% vs. 46.2% (p = 0.01). GLAD in FN-HR CTX differed between pulmonologists and hematologists-oncologists (25.0% vs. 43.6%, p < 0.001) and in FN-IR CTX (38.1% vs 48.6%, p < 0.001). Comparing gynecologists with hematologists-oncologists, GLAD in FN-HR CTX was 86.2% vs. 82.5% (p = 0.15) and in FN-IR CTX 58.6% vs. 55.6% (p = 0.38). CART analysis split pulmonologists and other specialists, with the latter adhering more to GL (p < 0.001). Hematologists-oncologists and gynecologists trained more than 2 years in medical cancer therapy adhered more closely to GL than others (68.7% vs 46.2%, p < 0.001). Pulmonologists attending 0-1 national congresses adhered more to guidelines than other pulmonologists (44.8% vs 24.3%, p < 0.001). Adherence to G-CSF GL in Germany increased but is still insufficient. Certified and comprehensive cancer centers show a higher rate of GL implementation. There is still a disparity between cancer types and between specialized oncologists. AIO-Studien-gGmbH AIO-SUP-0215, Grants: Amgen, Hexal Disclosure: Hartmut Link: Advisory Role: Amgen, Chugai, Hexal, Sandoz, Teva; Financing of Scientific Research: Amgen, Chugai, Hexal, Sandoz, Teva; Expert Testimony: Amgen, Hexal, Teva Petra Ortner: Financing of Scientific Research: Publikationshonorare von Amgen, Teva P949

Prophylaxis of chemotherapy-induced neutropenia with Lipegfilgrastim in patients with lung cancer: results from an interim analysis of the non-interventional study NADIR Potthoff K.1, Lück A.2, Frost N.3, Losem C.4, Weide R.5, Schulz H.6 iOMEDICO AG, Freiburg, Germany, 2Zentrum für Urologie und Onkologie, Rostock, Germany, 3Charité Campus Virchow-Klinikum, Berlin, Germany, 4Praxis für Onkologie und Hämatologie, Neuss, Germany, 5Praxisklinik für Hämatologie und Onkologie Koblenz, Koblenz, Germany, 6Praxis Internistischer Onkologie und Hämatologie (PIOH), Frechen, Germany 1

Introduction: Platinum-based chemotherapy regimens are the standard treatment for patients with lung cancer. Neutropenia is one of the most serious hematological and major dose-limiting toxicities that can comprise treatment outcomes. Guidelines recommend prophylactic granulocyte colony-stimulating factor (G-CSF) use for patients (pts) at high risk for febrile neutropenia (FN). Lipegfilgrastim (LIP) is a glyco-pegylated G-CSF approved to reduce the duration of neutropenia and the incidence of FN. Here we report on results from an interim analysis of the non-interventional study NADIR in pts with lung cancer. Methods: The prospective multicenter non-interventional study NADIR is conducted in 270 outpatient centers and hospitals across Germany, aiming to collect data on prophylactic LIP use in 2500 pts with different tumor entities under antineoplastic treatment in routine clinical practice. The objective of the study is to assess the effectiveness of LIP by determining the incidence of neutropenia grade 3/4 and FN. Results: At the time of data cut-off (3/2016), 2422 pts were enrolled by 198 sites. 1556 pts were evaluable, thereof 172 pts with lung cancer (NSCLC: 73; SCLC: 94). Mean age was 65.1 years for lung cancer pts, 56.4% of pts were male. At inclusion 68.0% of pts had an ECOG of 0 or 1 (14.5% missing data). In 79.1% of pts chemotherapy was applied in a palliative setting. 78.5% of pts were treated with platinum-based combination chemotherapy. 589 of 636 documented chemotherapy cycles (92.6%) were supported by LIP. Neutropenia grade 3/4 occurred in 33.1% of pts, one patient developed FN. Dose reductions were reported in 73 of 636 cycles (11.4%), in 10 cases due to chemotherapy-induced neutropenia. For 12.2% of pts LIP-related adverse events were reported. The most frequent LIP-related adverse event reported was leucocytosis (3.5%). LIP-related serious adverse events were reported for 1.7% of pts. Conclusions: In pts with lung cancer treated with a platinum-based combination regimen LIP was effective and well tolerated. The incidence of

Abstracts

neutropenia grade 3/4 and FN was low and comparable to literature. FN only occurred in < 1% of pts. Disclosure: No conflict of interest disclosed. P950

Prophylaxis of chemotherapy-induced neutropenia with Lipegfilgrastim in patients with Non-Hodgkin-Lymphoma (NHL): results from an interim analysis of the noninterventional study NADIR Fietz T.1, Wolff T.2, Schulz H.3, Sandner R.4, Reichert D.5, Hurtz H.-J.6, Müller J.7, Grebhardt S.7 Praxis für Innere Medizin, Hämatologie und Onkologie, Singen (Hohentwiel), Germany, 2Onkologie Lerchenfeld, Hamburg, Germany, 3Praxis Internistischer Onkologie und Hämatologie (PIOH), Frechen, Germany, 4Onkologische Praxis Passau, Passau, Germany, 5Gemeinschaftspraxis für Hämatologie und Onkologie, Westerstede, Germany, 6Onkologische Gemeinschaftspraxis, Halle (Saale), Germany, 7iOMEDICO AG, Freiburg, Germany 1

Introduction: Chemotherapy-induced neutropenia is the most serious hematologic toxicity of chemotherapy regimens, associated with the risk of life-threatening infections as well as chemotherapy dose reductions and delays that may comprise treatment outcome. Therefore, guidelines recommend prophylaxis with granulocyte colony-stimulating factor (G-CSF) for patients (pts) at risk for febrile neutropenia (FN). Lipegfilgrastim (LIP) is a glyco-pegylated G-CSF approved to reduce the duration of neutropenia and the incidence of FN. Here we report on results from an interim analysis of the non-interventional study NADIR in pts with NHL. Methods: The prospective multicenter non-interventional study NADIR is conducted in 270 outpatient centers and hospitals across Germany aiming to collect data on prophylactic LIP use in 2500 pts with different tumor entities under antineoplastic treatment in routine clinical practice. The objective of the study is to assess the effectiveness of LIP by determining the incidence of neutropenia grade 3/4 and FN. Results: At the time of data cut-off (3/2016), 2422 pts were enrolled by 198 sites. 1556 pts were evaluable, 205 of the evaluable pts were diagnosed with NHL. Mean age was 64.9 years for NHL pts, 55.1% of pts were male. At inclusion 81.0% of pts had an ECOG of 0 or 1 (13.7% missing data). Most pts were treated with (R)-CHOP (32.2%), (R)-CHOP-like (21.5%) and (R)-Bendamustin (12.2%). 941 of 990 documented chemotherapy cycles (95.1%) were supported by LIP. Neutropenia grade 3/4 occurred in 37.1% of pts, 2.0% developed FN grade 3. Dose reductions were reported in 70 of 990 LIP supported cycles (7.1%), in 7 cases due to chemotherapy-induced neutropenia. For 14.1% of pts LIP-related adverse events were reported. The most frequent LIP-related adverse event was bone pain (4.4%). LIP-related serious adverse events were reported for one patient. Conclusions: Our results confirm the effectiveness and safety of LIP in a real-life setting in NHL pts treated with (R)-CHOP-based chemotherapy. The incidence of FN and neutropenia grade 3/4 was low and in line with previous findings, leading to a remarkable high adherence to the scheduled chemotherapy protocol. Disclosure: Thomas Fietz: Financing of Scientific Research: TEVA; Other Financial Relationships: Reisekostenerstattung durch TEVA Sina Grebhardt: No conflict of interest disclosed. P951

Employment of G-CSF during dose-dense chemotherapy in routine care Weide R.1, Feiten S.2, Chakupurakal G.1, Friesenhahn V.2, Kleboth K.2, Köppler H.1, Lutschkin J.2, Thomalla J.1, van Roye C.1, Heymanns J.1 Praxisklinik für Hämatologie und Onkologie, Koblenz, Germany, 2Institut für Versorgungsforschung in der Onkologie, Koblenz, Germany 1

Introduction: Evaluation of the employment of G-CSF to deliver dosedense chemotherapy in routine care 2008 - 2015.

Oncol Res Treat 2016;39(suppl 3):1–294

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Methods: All patients who received dose-dense chemotherapy in an oncology group practice between 2008 - 2015 were analysed retrospectively concerning treatment application and the type of G-CSF-usage. Results: 190 patients with a median age of 54 (18 - 77) received 198 dosedense chemotherapies, consisting of 846 cycles. 69 (36%) were female, 121 (64%) were male. Intention of treatment was curative in 55% and adjuvant in 45% of therapies. Curative protocols most frequently used were CAD (28%), BEACOPP(esc) (26%), R-CHOP-14 (13%) and CHOP-14 (11%). Adjuvant protocols most frequently used were ETC +/- Trastuzumab / Pertuzumab (43%), AC / EC-14 (29%), dtEC / Docetaxel (7%), TAC (7%) and Paclitaxel-14 (6%). G-CSF usage in the order of frequency was PEG-filgrastim in 34%, LIPEG-filgrastim in 29%, filgrastim in 27% and Lenograstim in 7%. 157 dose-dense chemotherapies were administered completely and could be evaluated in terms of delays in the protocol; of these 116 (74%) could be applied as planned without time delay. 23 therapies (12%) had to be discontinued for any reason (progress, toxicities). Febrile neutropenia occurred after 20 therapies (10%). No patient died as a cause of dose-dense chemotherapy. Conclusion: The majority of patients receive pegylated filgrastim during dose-dense chemotherapy in routine care. Dose-dense chemotherapy can safely be applied to outpatients and febrile neutropenia rate is low. Disclosure: No conflict of interest disclosed. P952

Prophylaxis of chemotherapy-induced neutropenia with Lipegfilgrastim in patients with breast cancer: results from an interim analysis of the non-interventional study NADIR Fietz T.1, Kurbacher C.M.2, Trarbach T.3, Salat C.4, Rezai M.5, Lorenz A.6, Niemeier B.3 Praxis für Innere Medizin, Hämatologie und Onkologie, Singen (Hohentwiel), Germany, 2Gynäkologisches Zentrum Bonn-Friedensplatz, Gynäkologie I (SP Gynäkologische Onkologie), Bonn, Germany, 3iOMEDICO AG, Freiburg, Germany, 4Gemeinschaftspraxis für Innere Medizin, Hämatologie u. Internistische Onkologie, München, Germany, 5Luisenkrankenhaus GmbH & Co. KG, Düsseldorf, Germany, 6Frauenarztpraxis Dr. Lorenz, Hildburghausen, Germany 1

Introduction: Anthracycline and/or taxane-based (A/T) chemotherapies (CTx) are among the most effective treatments in breast cancer (BC). Many modern A/T regimens used in BC including dose-dense (dd) protocols with treatment intervals of ≤ 2 weeks are associated with a significant incidence of febrile neutropenia (FN) thus forcing a primary prophylaxis using granulocyte colony-stimulating factors (G-CSF). Lipegfilgrastim (LIP) is a glyco-pegylated G-CSF approved to reduce the duration of neutropenia and the incidence of FN. In 2014, a large-scaled non-interventional study (NIS) was initiated to obtained detailed information on the value of LIP in order to prevent both FN and severe neutropenia. Here we report on results from an interim analysis of the NIS NADIR focusing on the subset of pts with BC. Methods: The prospective multicenter NIS NADIR is conducted in 270 outpatient centers and hospitals across Germany aiming to collect data on prophylactic LIP use in 2500 pts with different tumor entities subjected to CTx in the clinical routine. The objective of the study is to assess the effectiveness of LIP by determining the incidence of neutropenia grade 3/4 and FN. Results: At the time of data cut-off (3/2016), 2422 pts were enrolled by 198 sites. 1556 pts were evaluable; 741 pts were diagnosed with BC, of whom 274 were treated with dd regimens. Mean age was 54.6/ 51.4 years for all BC pts and pts with dd regimens, respectively. In 89.0/ 96.7% of pts, CTx was applied in an adjuvant setting. Overall 96.1% of pts were treated with regimens containing A/T. 94.6/ 96.1% of documented CTx cycles were supported by LIP. Neutropenia grade 3/4 occurred in 29.4/ 33.9% of pts, 2.2/ 1.8% developed FN. Dose reductions were reported in 5.5%/ 4.2% of cycles, in 0.7%/ 0.6% of cases due to CTx-induced neutropenia. For 26.7/ 34.3% of pts, LIP-related adverse events (AE) were reported. The most

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frequent LIP-related AE was bone pain (11.1/ 18.6%). LIP-related serious adverse events were reported for 2.6/ 3.3% of pts. Conclusions: LIP was effective and well tolerated in BC pts treated with A/T as well as in the subgroup of pts with dd regimen. The low incidence of neutropenia grade 3/4 and FN was comparable to previous publications. Disclosure: Thomas Fietz: Financing of Scientific Research: TEVA; Other Financial Relationships: Reisekostenerstattung durch TEVA Beate Niemeier: No conflict of interest disclosed. P953

NEPA as antiemetic prophylaxis in cancer patients receiving highly or moderately emetogenic chemotherapy: Patientreported quality of life and efficacy Karthaus M.1, Rauh J.2, Guth D.3, Heilmann V.4, Schilling J.5 Klinikum Neuperlach, Department of Haematology and Oncology, München, Germany, 2Gemeinschaftspraxis Innere Medizin, Witten, Germany, 3 Gynäkologische Praxis, Plauen, Germany, 4Praxis Günzburg, Günzburg, Germany, 5Gynäko-onkologische Gemeinschaftspraxis, Berlin, Germany 1

Introduction: NEPA (Akynzeo®) is a fixed combination capsule that combines the new NK1-receptor-antagonist netupitant and the 5-HT3-receptor-antagonist palonosetron. It has been approved for the prevention of acute and delayed chemotherapy-induced nausea and vomiting (CINV) in adult cancer patients (pts) receiving cisplatin-based highly emetogenic chemotherapy (HEC) or moderately emetogenic chemotherapy (MEC). The objective of the study is to evaluate the quality of life of cancer pts undergoing MEC or HEC and receiving NEPA for CINV prophylaxis and to investigate the efficacy and safety of NEPA under real life conditions. Methods: This non interventional study is planned to evaluate 2500 pts receiving single day or two day MEC or HEC treated in > 100 German centers. NEPA is prescribed in accordance with the terms of the marketing authorisation. The primary endpoint is the pts´ quality of life as recorded by FLIE questionnaires. Secondary endpoints include efficacy, additional medication, safety data and AEs as documented by online questionnaire and patient diary. 3 consecutive chemotherapy cycles are documented. For documentation treating physicians use the ODM QuaSi® online documentation system. Results: At the time of abstract submission, 527 pts had been included. The majority of pts (352) had breast cancer. Median age was 56 years. 92% of pts had an ECOG status of 0 or 1. Most pts received (neo)adjuvant chemotherapy, only 22% were treated with palliative intent. In cycle 1, 407 pts were evaluable for efficacy (343 in cycle 2 and 288 in cycle 3). Efficacy was reported very good in cycle 1, 2 or 3 in 55,8%, 56,6% and 57,6% respectively, while this was reported good in an other 35,4%, 36,7% and 34,7% in cylce 1,2 or 3. Efficacy war reported satisfactory in cycle 1, 2 or 2 in 6,9%, 5,3% and 6,9% while it was documented poor in 2%, 1,5% and 0,7% only from cycle 1 to 3. Patient reported outcome as recorded by FLIE questionnaires and patient diaries has not been evaluated yet. Conclusions: The trial is in ongoing. In more than 90% of pts the efficacy of the antiemetic prophylaxis was rated as very good or good in all 3 chemotherapy cycles by the treating physician. Additional data on efficacy as well as quality of life will be presented at the meeting. Tab. 1.

Efficacy antiemetic prophylaxis Very good Good Satisfactory Poor

Cycle 1

Cycle 2

Cycle 3

27 (55,8%) 144 (35,4%) 28 (6,9%) 8 (2%)

194 (56,6%) 126 (36,7%) 18 (5,3%) 5 (1,5%)

166 (57,6%) 100 (34,7%) 20 (6,9%) 2 (0,7%)

Disclosure: Meinolf Karthaus: Advisory Role: Riemser, Helsinn; Financing of Scientific Research: Riemser, Helsinn J. Schilling: Financing of Scientific Research: Riemser

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P954

PSM peptides of Staphylococcus aureus enhance the p38CREB signaling pathway in dendritic cells induced upon TLR activation, thereby modulating cytokine production and T-cell priming Autenrieth S.E.1, Richardson J.1, Klenk J.1, Günter M.1, Kretschmer D.2, Pöschel S.3, Schenke-Layland K.3, Kalbacher H.4, Clark K.5, Armbruster N.1 Universität Tübingen, Medizinische Klinik, Innere Medizin II, Tübingen, Germany, 2Interfakultäres Institut für Mikrobiologie und Infektionsmedizin, Tübingen, Germany, 3Universität Tübingen, Department für Frauengesundheit, Tübingen, Germany, 4Universität Tübingen, Interfakultäres Institut für Biochemie, Tübingen, Germany, 5University of Dundee, Dundee, United Kingdom 1

Introduction: The challenging human pathogen Staphylococcus aureus has highly efficient immune evasion strategies for causing a wide range of diseases, from skin and soft tissue to life-threatening infections. Phenol-soluble modulin (PSM) peptides are major pathogenicity factors of community-associated methicillin-resistant S. aureus strains. Previously, we showed that PSMs in combination with a TLR2 ligand from S. aureus induce tolerogenic dendritic cells (DCs) characterized by the production of high amounts of IL-10 and lack of proinflammatory cytokine production, reduced antigen uptake and DC maturation. This in turn promotes the activation of regulatory T cells while impairing the Th1 response. However, the underlying molecular mechanisms remain unknown. Methods: Here, we addressed the effects of PSMs on signaling pathway modulation downstream of TLRs by flow cytometry, imaging flow cytometry and ELISA. Results: TLR2 stimulation in combination with PSMα3 led to increased and prolonged phosphorylation of NF-κB, ERK, p38, and CREB in mouse bone marrow-derived DCs compared with single TLR2 activation. Furthermore, inhibition of p38 prevented IL-10 production, which in turn reduced the capacity of DCs to activate regulatory T cells. Interestingly, the modulation of the signaling pathways by PSMs was (1) independent of the activated TLR and (2) independent of the known receptor for PSMs, as shown by experiments with DCs lacking the formyl peptide receptor 2. Instead, PSMs penetrate the cell membrane most likely by transient pore formation. Moreover, co-localization of PSMs and p38 was observed near the plasma membrane in the cytosol indicating a direct interaction. Conclusion: Thus, PSMs from S. aureus directly modulate the signaling pathway p38-CREB in DCs, thereby impairing cytokine production and in consequence T-cell priming to increase the tolerance towards the pathogen. Disclosure: No conflict of interest disclosed.

The cocci were then analysed by MALDI-TOF MS and indeed classified as Leuconostoc lactis. Leuconostoc lactis is a grampositive coccus found ubiquitously in the environment. It plays a role in the fermentation processes of milk products and in the production of wine. It is no hazard for the immunocompetent, but its pathogenicity has been described in the immunocompromised: it may cause endocarditis, osteomyelitis, bloodstream infection etc. The main entrances are skin and gastrointestinal tract. Isolates are often misdiagnosed as viridans streptococci or enterococci since it is not always correctly identified by automatized identification systems of bacteria. All grampositive bacteria from normally sterile compartments should therefore undergo a testing for resistance. Vancomycin resistance is rare in gram positive cocci and should primarily prompt exclusion of Vancomycin-resistant Enterococcus faecium, Leuconostoc spp. and Pediococcus spp. The suspicion may be raised by the microbiologist or by the attending physician. First choice antibiotics for the treatment of leuconostoc infections comprise penicillin, ampicillin, clindamycin. Cephalosporins and tetracyclines may also be effective, however the microbes are intrinsically resistent against vancomycin and teicoplanin. In conclusion, Leuconostoc lactis is a microbe of growing importance particularly in the immunocompromised patient. The identification requires a profound knowledge of the microbiologist as well as a tight cooperation between him and the clinician. Penicillin is the treatment of choice. Disclosure: No conflict of interest disclosed. P956

Utilisation of facilitated subcutaneous immunoglobulin treatment (HYQvia) in patients with SID: Preliminary data from the SIGNS registry Reiser M.1, Fasshauer M.2, Borte M.2, Baumann U.3, Gold R.4, Pittrow D.5, Huscher D.6, Stangel M.7, Sommer C.8, Otto G.P.9, Hensel M.10 Praxis Internistische Onkologie, Köln, Germany, 2Krankenhaus St. Georg, Leipzig, Germany, 3Klinik für Pädiatrische Pneumologie, Allergologie und Neonatologie, Medizinische Hochschule, Hannover, Germany, 4Klinik für Neurologie, Unversitätsklinikum, Bochum, Germany, 5Institut für Klinische Pharmakologie, Technische Universität, Dresden, Germany, 6Deutsches Rheumaforschungszentrum, Epidemiologie, Berlin, Germany, 7Klinik für Neurologie, Medizinische Hochschule, Hannover, Germany, 8Neurologische Universitätsklinik, Würzburg, Germany, 9Medizinische Abteilung, Baxalta GmbH, Oberschleissheim, Germany, 10Mannheimer Onkologie Praxis, Mannheim, Germany 1

Introduction: In neutropenic patients fever is common. Often no microbe can be isolated and an empirical antibiotic regimen is initiated. In positive blood cultures the most common microbes isolated are part of the patient´s endogenous flora, most of them gram-positive cocci. Occasionally microbes not yet known as pathogenic are found. Case report: We describe a patient suffering of acute myelogenous leukemia. After the second induction chemotherapy she developed neutropenic fever as well as colitis. An antibiotic treatment was started with ceftazidime, followed by meropenem. When grampositive cocci in short chains were isolated in blood cultures vancomycin was added without effect. The cocci were first misidentified as viridans streptococci, later as enterococci. Because of the continuous fever despite vancomycin a Leuconostoc infection was suspected and penicillin added resulting in prompt defervescence.

Patients with secondary immunodeficiencies (SID) receive human immunoglobulin (IgG) preparations intravenously (IV) or subcutaneously (SC) as replacement therapy to prevent severe infections. HyQvia is a new combination product whereby human hyaluronidase is injected SC (to temporarily enabling large volumes of drug solutions to be infused) followed by human IgG 10% infused through the same needle. This preparation allows fast delivering the full monthly IgG dose in one SC infusion every 3-4 weeks. We aimed to collect data of patients with SID on HyQvia from the prospective longitudinal non-interventional SIGNS study in Germany (NCT01287689). The infection rates and through levels were obtained. The abbreviated Treatment Satisfaction Questionnaire for Medication (TSQM-9) was used 6 months after treatment initiation in HyQvia patients. Of the 13 SID patients, 3 had chronic lymphatic leukemia (CLL), 4 Non-Hodgkin lymphoma of the B-cell type, 3 multiple myeloma, 1 indolent lymphoma, 1 leukemia (other than CLL) and 1 chemotherapy induced immune deficiency. Overall, mean age of patients was 58.8 ± 13.7 years, 69.2% were males. Mean body mass index was 25.9 ± 2.1 kg/m2. Mean disease duration since first symptoms was 7.3 ± 5.3 years and mean time since diagnosis 7.2 ± 5.3 years. Four patients received HyQvia as their first IgG preparation, while 7 (53.8%) had been prescribed other IgG before (6 IV, 1 SC) and were

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P955

Leuconostoc lactis – a rare microbe in neutropenic patients Thomssen H.1, Peyn A.1, Thilo N.2 Klinikum Bremen Mitte, Med. Klinik 1, Bremen, Germany, 2Laborzentrum Bremen, Bremen, Germany 1

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switched. For 2 patients the prior treatment was unknown. Treatment with HyQvia started with 4-week doses with a mean of 333 mg/kg body weight, and application intervals varied between 1 and 4.3 weeks. The mean IgG trough level at initiation of HyQvia was 6.6 ± 5.6 g/l. Infections in the 12 months prior to initiation were reported in 57% of pretreated patients and 2 patients of the 4 IgG-naïve patients (50%). After HyQvia initiation, so far no serious infections occurred, 33% of patients had general infections in the following 6 months. Treatment was overall well tolerated. In the 3 patients in whom TSQM-9 data were so far available at 6 months, global satisfaction was rated with 89.4 points, effectiveness with 90.7, and convenience with 64.8 (100 indicating highest satisfaction on each scale). The current preliminary analysis suggests that HyQvia as a new option for facilitated SC IgG treatment is well received by patients, and is effective in controlling infections in naïve patients or those switched from other IV or conventional SC IgG preparations. Disclosure: Marcel Reiser: Financing of Scientific Research: Baxalta GmbH Manfred Hensel: Financing of Scientific Research: Baxalta GmbH P957

Effectiveness of tigecycline (TGC) in hematological patients (pts) with persisting or recurring fever: a retrospective, monocentric review of 78 cases Geßner D.1, Deubrecht S.1, Fischer T.1, Schalk E.1 Otto-von-Guericke University Magdeburg, Medical Center, Department of Hematology and Oncology, Magdeburg, Germany 1

Introduction: Infections remain one of the leading causes of treatment-related deaths after chemotherapy in pts with hematological malignancies. Persisting fever, despite changing the initial antibiotic regimen, is an important clinical sign of ongoing infection. TGC, a broad-spectrum antibiotic without activity against Pseudomonas aeruginosa, is used in some centers to treat pts with fever unresponsive to the initial treatment. The aim of this study was to evaluate the effectiveness of TGC in pts with persisting or recurring fever. Methods: Records of pts with persistent or recurring fever who were treated with TGC in our department were retrospectively reviewed. Response rates and 30-day mortality were evaluated. Successful response was defined as defervescence (≥7 days) starting at least 5 days after beginning of TGC treatment. Neutropenia was defined as white blood cell count < 1.0 G/L or absolute neutrophile count < 0.5 G/L. Fever was defined as a temperature of ≥38°C. Results: 78 cases (median age 61 years, range 20-82; 54% men) of pts treated with TGC in our department from 07/2009 to 07/2015 were reviewed. 74 (95%) of the pts were treated for hematological malignancies, mostly acute myeloid leukemia (55%). 51 (65%) were neutropenic at the beginning of TGC treatment. 33 (42%) pts had radiologically diagnosed pneumonia, 22 (28%) microbiologically documented infection (MDI) and 23 (30%) fever of unknown origin (FUO). In 77 (99%) cases TGC was combined with another broad-spectrum antibiotic effective against P. aeruginosa, mostly ceftazidime (82%). 15 (19%) pts received TGC as second-line and 60 (77%) as third or fourth-line therapy. The overall response rate was 72%. The response rate was not significantly different for neutropenic vs. non-neutropenic pts (67% vs. 82%; p = 0.20). There was no difference in response rates regarding the type of infection (pneumonia, MDI, FUO: 73% vs. 73% vs. 70%, respectively). The overall 30-day mortality rate was 12% (10/78); 8 deaths were related to uncontrolled infection, 2 were related to progress of the underlying disease. There was no significant difference regarding neutropenic vs. non-neutropenic pts (16% vs. 7%; p = 0.48). Pts who did not achieve defervescence had an increased 30-day mortality rate (27% vs. 7%; p = 0.03). Conclusions: In our experience, TGC in combination with a P. aeruginosa-effective antibiotic may be an alternative treatment for hematological pts with persistent or recurring fever.

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Disclosure: Daniel Geßner: Other Financial Relationships: Reisekostenerstattungen Pfizer Pharma GmbH Enrico Schalk: No conflict of interest disclosed. P958

Prolonged Hepatitis-E-infection under rituximab-therapy Klank D.1, Zinke F.1, Weiß B.1, Hoffmann M.1, Uppenkamp M.1 Klinikum der Stadt Ludwigshafen, Medizinische Klinik A, Ludwigshafen, Germany 1

Since the 70ies, it is well known, that besides the Hepatitis-A-virus, another fecal-oral transmitted virus exists. Current statistics show, that 17 percent of the german population have undergone a Hepatitis-E-infection. Being asymptomatic in healthy people, a hepatitis-E-infection might not be controlled immunologically by immunosuppressed patients. The patient was admitted to our clinic in order to induce an immunochemotherapy after a Diffuse Large Cell Non-Hodgkin-lymphoma (DLCNHL) was being diagnosed. After the second cycle of CHOP14 and Rituximab the patient had an elevation of liver enzymes up to 8 times above the usual. Therefore the immunochemotherapy could not be continued. A further serologic testing for HIV and Hepatitis A, B, C and E showed no results. An autoimmune hepatitis, haemochromatosis and Wilson’s disease as cause of this elevation of liver enzymes could be ruled out. A liver punction brought no further information. Liver enzymes being stable, immunotherapy with Rituximab was continued. Due to a new rise of liver enzymes the third cycle of CHOP could not be given. In the following days an acute Hepatitis-E-infection was diagnosed by polymerase chain reaction (PCR) as cause of these elevated liver enzymes. We started an antiviral treatment with Ribavirin (1000mg/day orally), in order to prevent an imminent liver failure and chronification. Short time after beginning the Ribavirin-therapy a major drop of the liver enzymes could be measured and 4 weeks later Hepatitis-E-PCR was negative. The Ribavirin therapy was continued for 3 months. Consequently, a diagnosis of unclear elevation of liver enzymes should always include Hepatitis-E-testing. In immunosuppressed patients this testing must be performed by PCR as serological tests could be false negative. Disclosure: No conflict of interest disclosed. P959

Successful treatment of neutropenic MRSA bacteremia with septic superior vena cava thrombus and cerebral embolism using high dose daptomycin Schrenk K.G.1, Frosinski J.2, Scholl S.1, Otto S.3, La Rosée P.1,4, Hochhaus A.1, Pletz M.W.2 Klinik für Innere Medizin II, Hämatologie und Internistische Onkologie, Universitätsklinikum Jena, Jena, Germany, 2Zentrum für Infektionsmedizin und Krankenhaushygiene, Universitätsklinikum Jena, Jena, Germany, 3Klinik für Innere Medizin I, Kardiologie, Angiologie, Pneumologie und Internistische Intensivmedizin, Universitätsklinikum Jena, Jena, Germany, 4Klinik für Innere Medizin II, Onkologie, Hämatologie, Immunologie, Infektiologie und Palliativmedizin, Schwarzwald-Baar Klinikum, Villingen-Schwenningen, Germany 1

Patients with prolonged neutropenia after remission inducing chemotherapy in acute myeloid leukemia are at high risk for serious and potentially lethal infections. Here we present the successful treatment of a neutropenic patient with acute myeloid leukemia, MRSA bacteremia and cerebral embolism using high dose daptomycin. We discuss important aspects in the diagnostic and therapeutic approach in this setting. In 2012 a 49-year old woman was diagnosed with acute myeloid leukemia. After induction chemotherapy bone marrow aspiration revealed blast persistence and reinduction chemotherapy was started four weeks after induction chemotherapy. Three days after the end of subsequent consolidation chemotherapy, the patient suffered from fever up to 38,6°C. Two

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days after the first fever episode, blood culture demonstrated methicillin-resistant Staphylococcus aureus (MRSA) bacteremia. The patient deteriorated with development of somnolence. MRI scan revealed multiple cerebral lesions consistent with septic embolism. Transesophageal echocardiography (TEE) showed thrombus formation in the superior vena cava. As salvage therapy, the patient was started on high dose daptomycin 12mg/kg i.v., the twofold of the licensed dose of 6mg/kg. 12 hours after start of treatment with daptomycin the patient was awake and the fever had decreased. Follow-up imaging revealed a decrease of cerebral embolic lesions in MRI and the complete disappearance of the superior vena cava thrombus in the TEE. The presented case demonstrates several important aspects in the treatment of MRSA bacteremia in neutropenic patients. 1st, it shows the necessity of diagnostic workup with continuous monitoring of blood cultures. 2nd, the importance of correct choice and adequate dosage of antibiotics: daptomycin is a cyclic lipopeptide antibiotic with activity against gram-positive bacteria. In contrast to linezolid, daptomycin and rifampin have bactericidal activity and are very efficient in treating blood stream infections like endocarditis or septic embolism. The recommended dose of daptomycin is 4mg/kg for skin infections and 6mg/kg for Staphylococcus aureus bacteremia. Since the PK/PD parameter of daptomycin shows a concentration dependent killing, increased dosages up to 12mg/kg are now recommended for serious infections, particularly after failure of prior antibiotic treatment and persisting MRSA bacteremia. Publication: Ann Hematol. 2016 95(2): 355-357

fections decreased by up to 39.1% using octagam® 5% and by up to 41.7% using octagam® 10%. Conclusion: octagam® 5% and octagam® 10% are well tolerated in routine clinical use in SID patients. There is no relevant difference between both products concerning tolerability, ADRs occurred in 0.61% of the octagam® 5% infusions and in 0.58% of the octagam® 10% infusions.

Disclosure: No conflict of interest disclosed.

Introduction: CUP syndromes are malignant tumors which are characterized by metastases without an identification of the primary tumor. The incidence is 5 patient on 100,000 people with a mean age of 60yrs. 70% of these patients have a poor prognosis with a median survival of nine month. The therapy of the CUP syndromes is often a calculated, palliative and intensive antineoplastic chemotherapy which can lead to the development of a secondary AML with complex cytogenetic aberrations. Case report: We report about a 23yrs old man who had until his medical examination in February 2014 no remarkable medical history. The symptoms of the patient were fatigue and weakness. At this time point no further diagnostics were performed. One month later the patient developed intensive pain which led to further diagnostics in a clinic close to this home. A marked tumor was identified in the abdomen and also large bilaterally swelling of the lymph nodes. A lymph node dissection was performed. The histopathological analysis revealed an undifferentiated carcinoma. Therefore, we started a therapy with carboplatin and taxol. After the fifth course of chemotherapy the patient showed a progress of the disease. To the therapy VP 16 was added which led to a good response. Under this treatment the patient had a very good ECOG of 0 and was able to perform his works as shipbuilder. In November of 2015 a progressive thrombocytopenia and leukocytosis were observed. The diagnostics of the bone marrow revealed a myelomonocytic AML M5 after FAB. Following, a preliminary chemotherapy with Ara-C was initiated. Under this therapy a progress was observed. Therefore, the treatment was changed to mitoxantrone which showed a good response. After this therapy a treatment with decitabine was started. Unfortunately, the patient died due to an intracerebral bleeding. Conclusions: CUP syndromes are tumors which are not so rare. These tumors are characterized by a poor prognosis. Therefore, in the most cases a palliative therapy will be initiated. Secondary leukemias following antineoplastic therapy have a frequency of 5%. Due to the poor prognosis of the CUP syndromes the influence of the secondary leukemias on the overall survival is limited. In the presented case the response to the therapy of the CUP was better than usual.

P960

Tolerability and efficacy of the intravenous immunoglobulins octagam® 5% and octagam® 10% in secondary immunodeficiencies – Interim analysis of a non-interventional study Tschechne B.1, Jansen M.2, Klein I.2, Lietz C.2 Hämatologisch Onkologischer Schwerpunkt, Neustadt am Rübenberge, Germany, 2Octapharma GmbH, Langenfeld, Germany 1

Introduction: octagam® 5% and octagam® 10% are liquid human normal immunoglobulin solutions for intravenous administration (IVIG). They are indicated for replacement therapy in primary or secondary immunodeficiencies (PID or SID) and for immunomodulation in the treatment of autoimmune diseases. A prospective, open, multicentre non-interventional study (NIS) was started in 2014 in Germany to evaluate the tolerability of octagam® 5% and octagam® 10%. In this interim analysis we focus on the tolerability in patients with SID. Methods: Patients were treated according to the individual decision of their physician. All relevant data regarding the patient, the infusions with octagam® 5% and octagam® 10% and the occurence of adverse drug reactions (ADRs) were documented by using Case Report Forms (CRFs). Data received between January 1st 2014 to December 31th 2015 were included in this interim analysis. A subgroup analysis of SID patients is presented. Results: Data from 1,680 patients who received 20,150 infusions were collected in 158 sites. 1,350 patients (80%) received infusions because of immunodeficiencies, including 915 patients suffering from SID. The 915 SID patients received 10,695 infusions. 459 patients were treated with octagam® 5% and 456 patients were treated with octagam® 10%. This represents a mean ratio of 12 infusions per patient for both products. The median dosage per infusion was between 0.1 g/kg and 0.2 g/kg for octagam® 5% and 0.2 g/kg for octagam® 10%. For both products the median treatment interval was 4 weeks. ADRs occurred in 33 out of 5,414 infusions (0.61%) with octagam® 5% and 31 out of 5,281 infusions with octagam® 10% (0.58%). Efficacy was assessed in SID patients by analysing the annual infection rate before and during study. The mean annual infection rate for all in-

Abstracts

Disclosure: Barbara Tschechne: Financing of Scientific Research: Ja Christian Lietz: Employment or Leadership Position: Ja

Posterdiskussion

Kasuistiken – Infektionen und Onkologie P961

A case report: A CUP syndrome of a 23 years old man who was successfully treated with Carboplatin/Taxol und VP 16 led to a treatment associated secondary AML Gläser D.1, Lestin M.1, Freitag S.2, Jenzen D.1, Erichson A.1, Peters K.1, Krammer-Steiner B.1 Klinikum Südstadt, Hämatologie/Onkologie/Palliativmedizin/Hämostaseologie und Komplementärmedizin, Rostock, Germany, 2Universitätsklinkum Rostock, Medizinische Klinik III- Hämatologie/Onkologie/Palliativmedizin, Rostock, Germany 1

Disclosure: No conflict of interest disclosed.

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P962

Successful treatment of unresectable mucor mycosis of the scull base with combined local and systemic amphotericin B – a case presentation

MRI at diagnosis

Behlendorf T.1, Appel T.1, Lautermann J.2, Neef U.3, Schütte W.1 KH Martha-Maria Halle-Dölau, Klinik für Innere Medizin 2, Halle, Germany, KH Martha-Maria Halle-Dölau, Klinik für Hals-, Nasen- und Ohrenheilkunde, Halle, Germany, 3Gemeinschaftspraxis Hämatologie, Onkologie und Gastroenterologie, Halle, Germany 1 2

Introduction: A 76-year old male patient suffering from mantle cell lymphoma under third line treatment with ibrutinib was administered to our hospital due to massive headache, amaurosis of the left eye, paresis of the left Nervus facialis and left-sided trigeminus neuralgia. A MRI scan of the skull revealed a subcranial tumorous lesion. A biopsy was performed as other manifestations of the MCL were in remission. Histology showed an infiltration with mucor. Methods: Treatment was performed interdisciplinary by the departments for otorhinolaryngology and medical oncology. A local debridement could not resect all of the mucor infiltration due to close proximity to the sinus cavernosus. Systemic therapy with liposomal amphotericin B for 21 days was complemented with repeated instillation of conventional amphotericin B into the maxillary sinus and the placement of a sponge system with conventional amphotericin B in the resection cavern through the maxillary sinus. Results: Under combined treatment with liposomal amphotericin B and local application of conventional amphotericin B ct-scans could document a reduction of the tumorous mass although neurologic deterioration continued. After 3 weeks of i.v.-treatment the regimen was changed to an oral formulation of posaconazol on an outpatient base which was continued for another 3 weeks. Control MRI after another 4 months showed a complete remission of the subcranial lesion, visus is still impaired but neuralgia is completely resolved. Conclusion: In unresectable mucor lesions the combined treatment modalities of systemic and local application of amphotericin B seem to be suitable and safe. The high mortality of mucor infections in immunosuppressed patients in our opinion legitimates the exceptional treatment as described above.

Fig. 1. At diagnosis.

MRI after treatment

Fig. 2. After treatment.

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Disclosure: Timo Behlendorf: Other Financial Relationships: Reisekostenerstattung Gilead, MSD Wolfgang Schütte: No conflict of interest disclosed. P963

Severe skin toxicity associated with the new BCR Signaling Inhibitor Idelalisib Batereau C. , Vehling-Kaiser U. 1

Praxis Vehling-Kaiser, Landshut, Germany

Idelalisib is new targeted therapy for use as monotherapy or in combination with other agents to treat refractory or relapsed chronic lymphocytic leukemia (CLL) and indolent B-cell non-Hodgkin lymphoma (NHL). As oral agent it inhibit the B- cell receptor pathway. In clinical trials toxicities are described as usually mild and self-resolving and include hepatotoxicity, nausea, diarrhea and pneumonitis. Rash in a mild to moderate form occurred in up to 22% of all cases. We describe a heavily pretreated CLL-patient who developed a severe skin toxicity under treatment with Idelalisib. He showed a total body exanthema and a severe mucositis which led to discontinuation of the drug. Disclosure: No conflict of interest disclosed. P964

A case report of a patient with a chronic myeloid leukemia (CML) and development of blast crisis – correlation with new appeared genetic mutations detected by new generation sequencing Wittke C.1, Sklarz L.-M.1, Krohn S.1, Lück A.2, Glaeser D.1,3, Glaeser H.1, Große-Thie C.1, Freitag S.1, Junghanss C.1, Murua Escobar H.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany, 2Center for Oncology and Urology, Rostock, Germany, 3Clinic of Internal Medicine III – Hematology/ Oncology/Hemostaseology, Palliative Care, Complementary Medicine, Klinikum Südstadt Rostock, Rostock, Germany 1

Introduction: Since the approval of imatinib (IMI) and subsequently newer generation TKIs CML patients can be treated successfully aiming at a major molecular response. There we report clinical and molecular data of a patient that developed disease progression following a 10 year treatment with TKI. Case report: The male patient was diagnosed at the age of 50 years with CML chronic phase. Initial treatment consisted of IMI leading to a MMR. However 24 months later patient became resistant and bcr-abl transcripts increased. Cytogenetic and molecular analyses revealed a new mutation in the IDH gene and a second Philadelphia chromosome as well as +8. No resistance mutation was detected. Treatment was changed to nilotinib but dosage had to be reduced (400mg daily) due to headaches. Stem cell transplantation was declined by the patient. He achieved a MMR. At month +120 patient lost MMR and he was switched to dasatinib. However, dasatinib was not well tolerated and did not induce remission. Shortly after patient developed myeloid blast crisis. Cytosinarabinosid based AML induction chemotherapy was applied but unfortunately the disease was resistant. Therefore a novel induction scheme (azacitidine + bosutinib) was applied leading to rapid hematological CR. Patient received immediately a full matched allogeneic stem cell transplantation following a myeloablative conditioning (BuCy). Unfortunately the patient developed on day +21 severe infection complications and died on day +28. Conclusion: Development of a blast crisis is a rare event following TKI treatment. Molecular monitoring of bcr-abl transcripts by PCR allows detection of relapse. The application of NGS further allows the identification of novel additional mutations and guiding of therapy intensification i.e. allogeneic stem cell transplantation.

P965

Maintenance therapy with Nilotinib and ATRA after consecutive Dasatinib and AIDA-2009 induction therapies in an unfit elderly patient with newly diagnosed CML and promyelocytic blast crisis with a three way Philadelphia chromosome translocation Schön C.1, Haferlach C.2, Teleanu V.3, Ringhoffer M.1, Kündgen L.1, Schmier M.1, Bentz M.1 Städtisches Klinikum Karlsruhe, Medizinische Klinik III, Karlsruhe, Germany, MLL Münchner Leukämielabor, München, Germany, 3Universitätsklinikum Ulm, Innere Medizn III, Ulm, Germany 1 2

This case report describes the consecutive treatment regimens in a 64-year-old unfit male who presented with leucocytosis and splenomegaly with peripheral blood and bone marrow showing features of blast crisis chronic myeloid leukemia with blast cells of bizarre morphology. Cytogenetic analysis revealed (1) a variant Philadelphia chromosome translocation (t(9;22;14)(q34;q11;q22)), as seen in 5-10% of CML cases, and (2) a subclone harboring in addition a trisomy 8 and a t(15;17) (q24;q21) translocation, giving rise to the PML-RARA fusion gene, the genetic hallmark of acute promyelocytic leukemia (APL). Noteworthy, the patient neither showed signs of bleeding tendency nor exhibited elevated fibrin degradation products. Following commencement of cytoreductive hydroxycarbamide therapy, tyrosine kinase inhibitor (TKI) monotherapy with dasatinib 140mg daily was administered. Real-time quantitative PCR (RQ-PCR) initially showed treatment response, 8,4% to 1,3% for PML-RARA/ABL1 and 33,5% to 3,78% for BCR-ABL1/ABL1 (figure). However, due to a developing grade 4 thrombocytopenia and pleural effusion, Dasatinib dose reduction and discontinuation was necessary, leading to a rapid appearance of blast cells in the peripheral blood. Following standard induction chemotherapy with idarubicin 12mg/m² per day (days 2, 4, 6, 8) and ATRA 22,5 mg/m² twice daily for 60 days (AIDA-2009 protocol), the patient obtained a complete hematologic remission. Maintenance therapy was administered with increasing doses of Nilotinib, initially 150mg daily, then 300mg twice daily, and ATRA 22,5 mg/m² twice daily (days 1-14 every four weeks). Molecular genetic analysis for PML-RARA revealed nondetectable transcripts in both assays after 6 weeks of maintenance therapy (RQ-PCR and nested PCR), whereas BCR-ABL1 transcript levels remain stable (BCR-ABL1/ABL1: 3,7%). In summary, standard APL induction therapy, followed by TKI and ATRA is a feasible option for unfit elderly patients with chronic myeloid leukemia in PML-RARA positive blast crisis.

Fig. 1. RQ-PCR of BCR-ABL1 and PML-RARA. Disclosure: No conflict of interest disclosed.

Disclosure: No conflict of interest disclosed.

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P966

Exceptional causality of a Coombs-negative hemolytic anemia Krenosz K.J.1, Lenger D.1, Fuchs D.1, Fridrik M.A.1 Kepler University Hospital, Med Campus III, Department of Internal Medicine 3 - Hematology and Oncology, Linz, Austria 1

Acquired Coombs-negative hemolytic anemia is a rare disease. In December 2015 a 26 year old woman presented in our clinic with a macrocytic Coombs-negative hemolytic anemia and iron-deficiency (Hb: 92 g/l, MCV: 106.5 fl, reticulocyte: 6.94%, LDH: 264 U/l, haptoglobin: < 70 mg/l, PNH: neg., bone marrow aspirate and biopsy showed iron deficiency, confirmed by iron staining). Congenital hemolytic anemia was ruled out by peripheral blood smear. In 2009 she underwent a bariatric surgical procedure with a gastric bypass. In summer 2015 she had an i.v. infusion with 1.000 mg ferric carboxymaltose because of iron-deficiency anemia. She did not take regular oral multivitamin substitution. We administered 1.000 mg ferric carboxymaltose i.v. The anemia improved to Hb: 113 g/l, whereas hemolysis worsened. In March 2016 she presented in a worse general condition with muscle weakness, nausea, emesis, bone pain, severe hypalbuminemia and edema. Parenteral nutrition was administered, though clinical status and anemia (50 g/l minimum) decreased rapidly. Focusing on the reason for her condition and the Coombs-negative hemolytic anemia, we found a severe hypophosphatemia (inorganic phosphate: 0.22 mmol/l). With infusions of glucose-1-phosphate until normalization of serum phosphate the red blood cell count and the patient’s condition clearly improved within a few days. Ferric caboxymaltose infusion usually causes mostly asymptomatic, prolonged hypophosphatemia. In our patient this effect was intensified by the impaired resorption after bariatric surgery, and became life-threatening when parenteral nutrition caused a refeeding-syndrome with severe hypophospatemia. Phosphate substitution is mandatory after bariatric surgery and during refeeding. Phosphate levels should be controlled after ferric carboxymaltose infusions. Disclosure: No conflict of interest disclosed. P967

Launch of an hematology e-learning tool Thomalla J.1, Feiten S.2, Heymanns J.1, Köppler H.1, van Roye C.1, Weide R.1 Praxisklinik für Hämatologie und Onkologie, Koblenz, Germany, 2InVO-SOFT, Koblenz, Germany 1

Introduction: Medical education should combine theoretical sessions, practical exercises and individual elaborations of the learning matter. Hematology is complex and a lot of clinical experience is needed, especially with regard to diagnosis based on cytological results. For this purpose an hematology e-learning tool was launched in 2015. Methods: Personal experiences in dealing with the online tool zytologieseminar.de should be reported. Results: 247 case histories which cover entire hematology are presented. The tool can be used as a reference at the microscope, for exam preparation and for lesson planning. The user can switch between an interactive development of case histories and a lecturer mode. For each case medical history, laboratory parameters and 40-60 high-quality pictures of blood, bone marrow and lymph nodes as well as MRI or CT imaging are available. A search function allows a targeted search for disease patterns and a convenient download of the corresponding images. Numerous renowned hematologists enriched the collection by their own case reports. A scientific advisory board and an English translation will be available in the future. Experienced consultants have been using the tool for years to conduct hematology trainings for medical students.

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Conclusions: Zytologieseminar.de is a free software available to everyone who is interested in hematology and especially in cytology. It can be used as a lecturer, as a self-study tool or for reference purposes. Disclosure: No conflict of interest disclosed. P968

Acquired Pure Red Cell Aplasia in elderly patients with different underlying diseases: Report of 2 cases Glag S.1, Kanz L.1, Weisel K.1 Medizinische Universitätsklinik Tübingen, Abteilung II für Onkologie, Hämatologie, Immunologie, Rheumatologie und Pulmologie, Tübingen, Germany 1

Introduction: Acquired Pure Red Cell Aplasia (PRCA) is a rare cause of anemia mostly of unknown cause, but may be associated with various underlying diseases. PRCA is described to be associated with large granular lymphocyte (LGL) leukemia, myelodysplastic syndrome (MDS), thymoma, viral infection or immune disorders. Methods: We report 2 cases of pure red cell aplasia. Results: Case 1: A 75-year old woman was diagnosed with anemia (Hb 6.7 g/dl). Bone marrow evaluation revealed a hypocellular marrow with a total lack of erythropoietic progenitors. There was no evidence for abnormal or monoclonal B- or T-cell clones. PET- CT scan excluded lymphoma and other suspicious lesions. Treatment was initiated with corticosteroids (prednisone 1 mg/kg /day) and after 4 weeks Hb values had almost normalised. Subsequently, prednisone was tapered to 7.5 mg/day. 2 years later, she presented with locally advanced pancreatic cancer, which was inoperable. Paraneoplastic origin of PRCA may be suspected occurring 2 years before clinically apparent carcinoma. Despite regular visits including ultrasound evaluation, early disease diagnosis was not possible. The patient died from her pancreatic cancer. Case 2: A 73-year old man was diagnosed with anemia (Hb 5.9g/dl) in an outside institution. Bone marrow evaluation showed a slightly hypocellular marrow with a total lack of erythropoietic progenitors. Chest-XRay, abdominal ultrasound, gastroscopy and colonoscopy showed normal results. The patient was referred to us and bone marrow evaluation was repeated. We saw a hypercellular marrow with only few erythropoietic progenitors and dysplasia in granulopoiesis and megakaryopoiesis. Flow cytometric immunphenotyping showed a CD57+ T-cell population representing a T-LGL clone. This was underlined by a monoclonal T-cell population demonstrated by T-cell-receptor rearrangement in the bone marrow biopsy. With a diagnosis of T-LGL, the patient was treated with corticosteroids (prednisone 1 mg/kg/day). After 4 weeks, Hb values were almost normalised and prednisone was reduced to 5 mg/day. 3 years later he still remains under our follow up. Conclusion: PRCA is a rare cause of anemia. Upon presentation, hematologic malignancies and solid tumors as the underlying cause have to be carefully excluded. However, as a paraneoplastic phenomenon PRCA might occur years before presentation of the underlying malignant disease. Treatment consists of steroids and/or treatment of the underlying cause. Disclosure: No conflict of interest disclosed. P969

Afatinib induced remission of a colorectal adenoma – a case report Gudzuhn A.1, Glitsch A.2, Schreiber A.2, Döring P.3, Spoerl M.4, Schmidt C.A.1 Universitätsmedizin Greifswald, Hämatologie/Onkologie, Greifswald, Germany, Universitätsmedizin Greifswald, Viszeral- und Thoraxchirurgie, Greifswald, Germany, 3Universitätsmedizin Greifswald, Institut für Pathologie, Greifswald, Germany, 4Universitätsmedizin Greifswald, Radiologie, Greifswald, Germany 1 2

Introduction: Afatinib is an established treatment in EGFR mutated adenocarcinoma of the lung showing activating EGFR-mutations. Under

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CONTENTS AUTHOR INDEX

evaluation is the use in the treatment of breast carcinoma and head and neck squamous cell carcinoma. However, little is known about its effectiveness in other tumor entities. Afatinib is a strong and selective inhibitor of all members of the erb-b- family of receptor tyrosine kinases, which are active in other tumor entities as well. We here report data from a patient treated with afatinib for advanced lung cancer showing surprisingly a distinct response of a simultaneous diagnosed rectal adenoma. Methods: In a 65-year-old patient with advanced EGFR mutated adenocarcinoma of the lung pretherapeutic workup showed a 3×2cm PET positive rectal mass. The tumor turned out to be a tubulovillous adenoma, extending to 4cm in rectoscopy (7-11cm ab ano). During treatment of the lung cancer with oral afatinib 40 mg per day, the rectal adenoma decreased in size to be finally resected at a size of 1cm by rectal wall excision. Results: Since no other tumor specific treatment was given, we speculate that the receptor-tyrosine kinase inhibitor afatinib is the cause of the tumor regression of the tubulovillous adenoma of the rectum. To our knowledge, this is the first report showing activity of afatinib in this tumor entity. Clinical data together with radiological and histological data will be presented. Conclusion: Further systematic research is required to evaluate afatinib as potential future treatment option in colorectal neoplasia. Disclosure: No conflict of interest disclosed.

Wissenschaftliches Symposium

Biologie und Therapie der indolenten Lymphome V970

Molecular risk stratification for follicular lymphoma Weigert O.1, German Low Grade Lymphoma Study Group (GLSG) Klinikum der Ludwig-Maximilians-Universität München, Campus Großhadern, Medizinische Klinik und Poliklinik III, Experimentelle Leukämie- und LymphomForschung (ELLF), München, Germany

failure of first-line immunochemotherapy and death, and could support clinical trials for alternative upfront regimens with highest antitumor activity, potentially accepting higher toxicity as deemed acceptable for the majority of patients with low-risk disease. Disclosure: No conflict of interest disclosed. V971

The genetic architecture in Waldenström´s Macroglobulinemia and its therapeutic implications Buske C.1 Universitätsklinikum Ulm, CCC Ulm, Klinik für Innere Medizin III, Ulm, Germany

We already know for longer time that Waldenström´s Macroglobulinemia (WM) is a molecularly heterogenous disease. However, just in recent years it was shown that WM is characterized by two mutations, which are not specific for WM, but allow to form three genotypes in this disease: the first mutation is the activating MYD88 mutation, which occurs in about 90% of the WM cases. The other mutation is an activating mutation in the CXCR4 gene, which can be detected in about every third patient with WM. Using the mutational status of these two genes patients are either MYD88mutated/CXCR4wildtype, MYD88mutated/CXCR4mutated or MYD88wildtype/ CXCR4wildtype. Of note, retrospective as well as prospective clinical data document that the group of patients with the MYD88wildtype/CXCR4wildtyp genotype respond poorly to therapy and show a significantly inferior response rate to ibrutinib. Ongoing clinical trials have to confirm the prognostic and predictive power of these genotypes and importantly, have to define alternative treatments which are able to overcome treatment resistance in MYD88wildtype/CXCR4wildtyp WM. Disclosure: Christian Buske: Financing of Scientific Research: Roche, Janssen; Expert Testimony: Roche, Janssen

Follicular lymphoma (FL) is among the most common lymphomas worldwide. It is a clinically and molecularly heterogeneous disease and remains incurable for most patients. Current immunochemotherapy regimens result in long-lasting remissions and excellent overall survival (OS) in the majority of patients who require systemic treatment. However, ~20% of patients experience short remissions and a median OS of < 5 years. Clearly, strategies to guide risk-adapted treatment approaches in FL are needed to avoid overtreatment of low-risk patients, and to prioritize alternative over standard regimens in high-risk patients. We analyzed the clinical data and the mutational status of 74 recurrently mutated genes from two independent cohorts: 151 patients from the GLSG2000 trial who received R-CHOP and IFN maintenance, and 107 patients from a population-based registry of the British Columbia Cancer Agency (BCCA) who received R-CVP followed by R-maintenance in 93 patients (87%). We developed an improved clinicogenetic risk model (m7-FLIPI) for failure-free survival (FFS) that includes the mutation status of 7 genes (EZH2, ARID1A, MEF2B, EP300, FOXO1, CREBBP, and CARD11), the FL International Prognostic Index (FLIPI), and the Eastern Cooperative Oncology Group (ECOG) performance status. A total of 43 GLSG (28%) and 24 BCCA patients (22%) were classified as high-risk by m7-FLIPI, with a 5-year FFS of 38% (vs 77%) and 25% (vs 68%), and a 5-year OS of 65% (vs 90%) and 42% (vs 84%), respectively (all p < 0.001). The m7-FLIPI is a promising starting point for upfront patient stratification by actual risk, but several challenges remain to be addressed, including the standardization of sequencing technologies and analysis pipelines to ensure widely reproducible results. Also, the m7-FLIPI and subsequent risk models will have to be validated in additional and larger cohorts, and evaluated in the context of specific treatments, in particular molecular targeted approaches, thereby ultimately paving the way from risk-adapted to biology-directed treatment algorithms. In conclusion, the m7-FLIPI currently represents the most promising predictor to identify the smallest subgroup of patients at highest risk of early

Abstracts

V972

BRAF in Hairy cell leukemia – treatment relevance? Zenz T.1 Molekulare Therapie in der Hämatologie und Onkologie / NCT / DKFZ / Uniklinik Heidelberg, Heidelberg, Germany 1

Targeted treatment approaches are transforming the therapeutic landscape of cancer care. The discovery of the BRAF V600E mutation in most cases of classical hairy cell leukemia opens up unique opportunities for tumor specific treatment of HCL targeting the MEK/ERK signaling pathway. In a rare disease as HCL, the oppportunities from cross entity (basket) approaches will leverage treatment opportunities in HCL. The discovery and biological implications of BRAF V600E in HCL are summarized to form a basis for our current understanding of the potential for clinical exploitation of BRAF V600E in HCL. There is overwhelming clinical evidence for activity of inhibitors of BRAF in the disease. The review will review current trial activity as well as discuss novel trial concepts exploiting targeted treatment focusing on BRAF inhibition in HCL. Disclosure: No conflict of interest disclosed. V973

Has the time come for chemotherapy-free therapy of follicular lymphoma? Hiddemann W.1, Hoster E.1, Schmidt C.1, Unterhalt M.1, GLSG LMU München, Med. Klinik III, München, Germany

The internationally widely accepted current standard for first-line therapy of follicular lymphomas at advanced stages III and IV consists of the combination of the antiCD 20 antibody Rituximab with chemotherapy comprizing either Bendamustin or CHOP for 6 cycles which is followed by a two year maintenance with Rituximab in responding patients.

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Based on rapidly increasing insights into the biology of follicular lymphomas and the characterization of crucial pathways regulating growth and differentiation, a variety of different agents has been developed which are currently explored in clinical phase I to III studies. Relatively mature data are already available for Lenalidomide and its combination with Rituximab which suggest that this combination may be equally effective to established Rituximab-chemotherapy combinations giving rise to the hope to replace cytotoxic regimens by chemotherapy-free treatment options in the near future. Other agents such as inhibitors of components of the B-cell receptor pathway such as Ibrutinib, or drugs interfering with apoptotic mechanisms such as Venetoclax as well as immune-checkpoint inhibitors have also shown promising activity and are currently explored in clinical validation trials. Further potentially effective agents are bispecific antibodies (BiTe) inducing a T cell attact against lymphoma cells but also antibodies coupled with toxins. Overall, the time has certainly come to explore chemotherapy-free combinations for first-line therapy of follicular lymphomas within the scope of clinical studies. However, the results of these trials need to be awaited before chemotherapy-free treatment options may enter broad clinical application outside of clinical studies. Disclosure: Wolfgang Hiddemann: Financing of Scientific Research: Roche, Gilead; Expert Testimony: Roche, Gilead, Janssen Michael Unterhalt: No conflict of interest disclosed.

Fortbildung

Kompetenznetz – Akute und chronische Leukämien V975

Methodological advances in quantification of minimal residual disease in ALL Brüggemann M.1 Sektion für Hämatologische Spezialdiagnostik, Klinik für Innere Medizin II, UKSH, Campus Kiel, Kiel, Germany 1

Monitoring of minimal residual disease (MRD) has become routine clinical practice in frontline treatment of childhood and adult ALL patients. The current gold standards for MRD quantification are the allele specific real-time quantitative PCR (ASO RQ-PCR) of clonal immunoglobuline (IG) and T-cell receptor (TR) gene rearrangements or multicolor flow cytometry, both with their own pitfalls and limitations. New high throughput tools like next generation flow, ddPCR and NGS might overcome a part of these drawbacks. Next generation flow cytometry: The EuroFlow consortium has introduced new high throughput concepts in flow-MRD, based on multidimensional data analysis. An important feature in the design of the MRD antibody tubes was the selection of antibody-fluorochrome combinations, that give insight in the full normal BCP pathway in BM, which allows to define the degree of immunophenotypic deviation of BCP-ALL cells from the normal pathway. New cell sample processing is introduced to reach sensitivities below 10-4 requiring fully standardized approaches, including instrument setting, sample processing with bulk lysis procedure, immunostaining, data acquirement, and data analysis with standardized gating strategies. ddPCR: ddPCR using ASO primers has the potential to generate an absolute read out that is largely tolerant of variations in PCR efficiency even for low target copy numbers, reducing the requirement for internal standardization. However, background amplification may limit sensitivity in a considerable proportion of assays. Prospective analyses of unselected cases have to be performed to verify the clinical impact of ddPCR-based MRD detection. Next generation sequencing of IG/TR gene rearrangements: For this approach, multiplex PCR V-, D- and J-primer sets are being used to amplify and sequence all potential rearrangements in a sample consecutively sequencing them with a high depth thereby avoiding the laborious design and testing of patient specific assays. Moreover, the readout is potentially

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more specific and sensitive than RQ-PCR. IG/TR NGS can also be used to detect evolved clonal IG/TR gene rearrangements and allows for additional insights into the background repertoire of B- and T-cells. Standardization, validation and quality control of the technology in a large-scale, multi-laboratory, scientifically controlled, multidisciplinary and independent setting are currently performed within a European collaboration, the EuroClonality NGS Network. Disclosure: No conflict of interest disclosed. V976

Chronic myeloid leukemia Lange T.1 Asklepios Klinikum, Weißenfels, Germany

Standard diagnostic procedures for chronic myeloid leukemia consist of clinical and hematological assessment of blood and bone marrow to define disease phase, calculate risk scores and monitor for hematological response. Beyond this, there is a need to assess cytogenetic response and perform molecular analyses to monitor BCR-ABL levels and determine mutation status in case of resistance or suboptimal response. Five tyrosine kinase inhibitors (TKI) have been approved in CML, three for first line treatment. The achievement of optimal response remains the major goal but treatment discontinuation after achievement of deep molecular response is now a realistic goal for a significant number of CML patients. Approximately 50% of patients will achieve this goal after 5 years of treatment with second generation TKI compared to 23% with imatinib 400mg. The rate of molecular relapses after discontinuation is again approximately 50% with the majority of relapses occurring within the first 6 months after stopping. Current study activities focus on the optimization of induction and maintenance as well as the discontinuation of TKI therapy in first line CML (i.e. TIGER Study) or in case of suboptimal response (i.e. Decline study). Further studies investigate the dose optimization of dasatinib (i.e. DasaHIT study), bosutinib (i.e. Bodo study) or ponatinib (Ponderosa study). The new ABL inhibitor ABL001 or the combination of nilotinib and ruxolitinib are currently available in a phase I study for resistant patients. Disclosure: Thoralf Lange: Financing of Scientific Research: Novartis, BMS; Expert Testimony: Novartis

Freier Vortrag

Ethik und Ökonomie V979

Importance of a good patient-communication in situations of treatment limitation – results from the EPAL-Study (Ethics Policy for Advanced Care Planning and Limiting Treatment) Mehlis K.1, Jaeger E.2, Laryionava K.1, Mumm F.2, Hiddemann W.2, Heußner P.2, Winkler E.C.1 Nationales Centrum für Tumorerkrankungen Heidelberg, Medizinische Onkologie Universitätsklinikum Heidelberg, Heidelberg, Germany, 2Klinikum der Universität München, Medizinische Klinik und Poliklinik III – Großhadern, München, Germany 1

Introduction: The aim of this study was to describe the extent to which advanced cancer patients are satisfied with the communication between them and their physician in situations of decisions to limit treatment (DLT) and to investigate factors that impact the perceived quality of patient-communication. Methods: This prospective quantitative study recruited 50 cancer patients from 5/2014 to 6/2015 in the Department of Hematology/Oncology at the University Hospital of Munich. Included were advanced cancer patients with treatment limitations either being discussed or decided. They completed a set of validated questionnaires on patients’ information needs, in-

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volvement in DLT, treatment goals and perceived quality of patient-communication using the questionnaire on the Quality of Physician-Patient Interaction (QQPPI). Results: The QQPPI total score was mean = 3.3/SD = 0.96 (range from 1=lowest quality to 5=highest quality). Those who report a good interaction felt significantly better informed about their diagnosis (p = 0.003), treatment options (p = 0.019), progress of the disease (p = 0.000) and side effects (p = 0.008) compared to those being less satisfied with the quality of physician-patient interaction. They also indicate significantly more often a shared decision-making together with their physician (p = 0.004), felt more involved in DLT (p = 0.017) and were more satisfied with the decision-making process (p = 0.010). Patients who believe that the treatment goal is cure rated the physician-patient interaction better (QQPPI mean = 3.9), compared to those believing cure is unlikely (mean = 3.1) and those being convinced that cure is not possible (mean = 3.7) (differences are not significant due to the small number of cases). Conclusion: Given the fact that the quality of physician-patient interaction is strongly related to the patients’ level of information, participation and satisfaction concerning treatment limitations, which are the very themes that facilitate DLT, there is a need to ensure good communication in these crucial decisions – e.g. by focusing on triggers for end-of-life communication. We aim for this in the EPAL-project with a policy for advanced care planning and limiting treatment. A post-implementation study has already started and will evaluate the impact of the policy. Disclosure: No conflict of interest disclosed. V980

Patients’ information needs and involvement in decisions to limit treatment in the EPAL-Study (Ethics policy for advanced care planning and limiting treatment) Mehlis K.1, Jaeger E.2, Laryionava K.1, Mumm F.2, Hiddemann W.2, Heußner P.2, Winkler E.C.1 Nationales Centrum für Tumorerkrankungen Heidelberg, Medizinische Onkologie Universitätsklinikum Heidelberg, Heidelberg, Germany, 2Klinikum der Universität München, Medizinische Klinik und Poliklinik III – Großhadern, München, Germany 1

Introduction: In order to inform the development of an institution-wide policy on decisions to limit treatment (DLT) this prospective quantitative study aims to investigate relevant factors for End-of-life decision-making with advanced cancer patients. Methods: We recruited 50 patients from 5/2014 to 6/2015 in the Dept. of Hematology/Oncology at the University Hospital in Munich. Included were only advanced cancer patients with treatment limitations either being discussed or already decided. They completed a set of validated instruments on patients’ information needs, decision-making preferences and treatment goals. Results: Patients report less information needs about prognosis and advanced directives than about diagnosis, treatment options and side effects. These information needs depend on the patients’ preferred role in decision-making: patients with an active role have higher information needs and are less often satisfied with their opportunities to get involved compared to those preferring shared decisions or delegating decisions to their physicians. 94% (n = 47) of the patients state that they agree with their physician on treatment goals. Still, they overestimate their chances for cure, as 15% of the patients say that the treatment goal is cure, 60% cure is unlikely and only 25% cure is not possible. 88% (n = 44) of the patients had a DNR and 78% (n = 39) a No-ICU order. While most of the patients (93%, n = 46) want to be involved in DLT, only 34% with DNR and 36% with No-ICU order confirm that they decided together with their physician over these limitations. Conclusion: Taking into account that we only included terminally ill cancer patients, some seem to have an unrealistic understanding of their disease. This might be caused by their low information seeking for prognostic information or their level of participation in the decisions on limiting

Abstracts

treatment. These results were fed into the development of an ethics policy for advanced care planning and limiting treatments. A post-implementation study started at the beginning of 2016 to evaluate the impact of the policy. Disclosure: No conflict of interest disclosed. V981

Patients’ preferences concerning quality and quantity of life – Results from the EPAL-Study (Ethics Policy for Advanced Care Planning and Limiting Treatment) Jaeger E.1, Mehlis K.2, Mumm F.1, Laryionava K.2, Hiddemann W.1, Winkler E.C.2, Heußner P.1 Klinikum der Universität München, Medizinische Klinik III, München, Germany, Nationales Centrum für Tumorerkrankungen (NCT), Heidelberg, Germany

1 2

Introduction: In cases of advanced cancer indicating the limitation of treatment, medical teams, patients, and their relatives are confronted with a number of clinical, ethical, and psychological conflicts. The identification of conflicts, the timing of consultations, and the effort of common decision-making in view of end of life should follow thoroughly investigated and generally approved guidelines integrating medical aspects, patient participation and patient´s information needs, as well as legal and ethical issues. A special focus of this data collection were patients’ preferences concerning quantity and quality of life and whether these preferences and their acknowledgement by doctors were considered into end-of-life decision making. Methods: Between May 2014 and July 2015, the guideline has been developed by representatives of all professional groups of the Department of Haematology/Oncology at the LMU Hospital (Munich). The accompanying study encompassed haematological/oncological patients (n = 50) with whom limitation of therapy had already been discussed or decided respectively. In addition, the opinions and attitudes of both doctors (n = 36) and nurses (n = 45) have been retrieved. For this purpose, a standardized questionnaire was applied to find out demographic data and the care-givers´ views of the patients´ quality of life and also both groups´ moral distress resulting from conflicts during the decision-making process leading to the limitation of therapy. Results: According to the survey, 57,2% (n = 28) of the patients prefer a better quality of life to quantity of life time. Only 10,2% of the patients (n = 5) wish to live longer even if this means forfeiting quality of life. 22,5% (n = 11) of the patients prefer a longer life time with no restrictions in terms of quality of life. Further analysis show that there is no correlation between patients’ and doctors’ rating concerning patient’s quality of life preferences. Conclusion: The majority of patients display a clear preference for maintaining the quality of life to its prolongation. In order to ascertain whether these findings are taken into account and in order to evaluate the guidelines´ impact on the attitude and behaviour of patients, doctors, and nurses in general, a post-implementation study has been started in January 2016. Disclosure: No conflict of interest disclosed.

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V982

To treat or not to treat? An empirical-ethical analysis of oncologists’ strategies on decision making in cases of optional adjuvant chemotherapy Haltaufderheide J.1, Schildmann J.1, Schmitt W.2, Uhl W.3, Vollmann J.1, Reinacher-Schick A.4 Ruhr Universität Bochum, Institut für Medizinische Ethik und Geschichte der Medizin, Bochum, Germany, 2St. Josefs-Hospital, Klinik für Allgemeine Innere Medizin, Bochum, Germany, 3St. Josefs-Hospital, Klinik für Allgemeinund Viszeralchirurgie, Bochum, Germany, 4St. Josefs-Hospital, Abteilung für Hämatologie und Onkologie, Bochum, Germany 1

Background: For many patients adjuvant chemotherapy according to guidelines is optional. In such situation oncologists and their patients need to exchange clinically relevant as well as value related information to make a decision for or against chemotherapy. In this empirical-ethical study we aim to reconstruct strategies used by oncologists and patients to facilitate these decisions with a focus on ethically relevant aspects. Methods: Non-participant observation of the decision making in patients with gastrointestinal tumor for whom adjuvant chemotherapy is optional according to current clinical guidelines. In addition we conducted semi-structured interviews subsequent to the observation. Observation notes and interview transcripts were analysed according to principles of grounded theory. Results: Up to present we have analysed data from 17 observations and 6 interviews. According to our data in the beginning of the consultation oncologists and physicians have different aims. While patients focus on medical information oncologists try to elicit clinical, biographical and value related information to be able to individualise the process of information giving. The observed oncologists use a range of strategies to elicit preferences and further information relevant to the decisions about optional chemotherapy. We identified two main types of strategies. Type one is focusing on biographical aspects of the patient and the establishment of a caring relationship. Type two is characterized by making explicit the uncertainty of the decision situation and a rational deliberation about the pros and contras of available treatment options. Regardless of the strategy used for facilitating the decision making we could observe situations in which patients and oncologists failed to reach an exchange of information and making a decision. Conclusion: Our research indicates that oncologists may benefit from a more systematic approach towards decision making in situations of optional chemotherapy and associated uncertainty. A multi-centre study with mixed-method approach should be conducted to elicit robust data which is necessary to provide well founded recommendations with regards to decision making in these situations. Disclosure: No conflict of interest disclosed. V984

Perceptions of and problems with the German DRG system, a representative survey of the DGHO Ostermann H.1, Krause S.2, Haag C.3 Klinik der Universität München, Medizinische Klinik und Poliklinik III, München, Germany, 2Universitätsklinik Erlangen, Medizinische Klinik 5, Erlangen, Germany, 3Universitätsklinikum Carl Gustav Carus an der Technischen Universität Dresden, Medizinische Klinik und Poliklinik 1, Dresden, Germany 1

Introduction: The German DRG system as a tool for diagnosis and procedure driven reimbursement of hospitalised patients was introduced in 2003. It is not known how the system is perceived by haematologists/oncologists and what problems are associated with its use. Methods: An online survey detailing questions of knowledge of the DRG system and hospital reimbursement was developed. Requests to participate were sent out in February 2016 to all members of the DGHO. Results: 11% of DGHO members completed the questionnaire. Two thirds of those were either department directors or attending physicians. About half of the participants reported to have good knowledge of the tools of

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the DRG system. In house reporting of reimbursement and costs was reported to be insufficient by 55% of participants. In 30% the cost calculation of the DRGs performed by the InEK was used to calculate the number of physicians necessary for the department. 28% of responders reported that money that should be allocated to research was misused to finance sick care. 90% regarded the possibility to reimburse especially expensive drugs besides the DRG lump sum (Zusatzentgelte, NUB) as important. In 20% of the hospitals more then 20% of the cases were scrutinized by physicians (MDK) on assignment of the sickness funds to find flaws in the documentation to possibly lower the invoiced sum of the DRG. In open questions some members seemed to be strictly opposed to the whole system, while most reported to be in need of more help to gain insight and understanding of the reimbursement system. Conclusion: 13 years after its implementation the German DRG system has in general been accepted by haematologists /oncologists. It seems to be necessary to improve knowledge of the system and of the ways hospitals report reimbursement and cost details to physicians. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Lungenkarzinome, Sarkome V985

Immunosequencing of circulating blood cells as response prediction biomarker in patients with solid tumors undergoing immune checkpoint inhibition with antibodies directed against PD-1 Brandt A.1, Akyüz N.1, Asemissen A.M.1, Schliffke S.1, Ford C.1, Radloff J.1, Stein A.1, Quidde J.1, Gökkurt E.2, Loges S.1, Haalck T.3, Mährle T.1, Bokemeyer C.1, Binder M.1 II. Medizinische Klinik Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany, 2Hämatologisch-Onkologische Praxis Eppendorf, Facharztzentrum Eppendorf, Hamburg, Germany, 3Ambulanzzentrum des Universitätsklinikum Hamburg-Eppendorf, Fachbereich Dermatologie, Hamburg, Germany 1

Cancer immunotherapy with antibodies targeting immune checkpoints, such as programmed cell death (PD-1), shows encouraging results, but reliable biomarkers predicting response to this costly treatment approach are lacking. In order to derive a potential immune signature predicting response to anti-PD-1 antibodies, we performed blood-based T- and B-cell repertoire profiling in 16 patients with solid tumors before and shortly after initiation of immune checkpoint inhibition by multicolor flow cytometry and deep immunosequencing. We found suppression of PD-1 positivity on all T-cell subsets after the first dose of the antibody. Furthermore, both T- and B-cell compartments quantitatively expanded during treatment. Expansions were mainly driven by an increase in the central memory, effector memory and activated T-cell compartments, as well as in all mature B-lineage compartments. Deep immunosequencing revealed a clear diversification pattern of the clonal T-cell space indicating antigenic selection in 47% of patients, while the remaining patients had stable repertoires. Half of the patients with a diversification pattern showed disease stabilization in response to the PD-1 inhibitor, while the remaining patients died shortly after initiation of treatment before the immune effect could have resulted in a clinical response. No disease stabilizations were observed without clonal space diversification. In restrospect, the low rate of disease stabilizations (19%) in this “real-life“ patient cohort indicates that the life span of patients in this study was often too short to derive benefit from immunotherapy, suggesting inadequate patient selection. As there is a strong need for predictive biomarkers in the setting of immune checkpoint inhibition we propose that immune repertoire analysis by immunosequencing of circulating T- and B-cells should be prospectively evaluated in the context of clinical studies.

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Disclosure: No conflict of interest disclosed. V986

Time to deterioration as measure for symptom burden and quality of life – Data from the LuLife project in 507 patients with non-small cell lung cancer in Germany von Verschuer U.1, Sandner R.2, Tessen H.W.3, Emde T.-O.4, Schnitzler M.5, Binninger A.5, Jänicke M.5, Marschner N.6 Hämato-Onkologische Gemeinschaftspraxis Dres. Rudolph & von Verschuer, Essen, Germany, 2Onkologische Praxis, Passau, Germany, 3Onkologische Schwerpunktpraxis, Goslar, Germany, 4Oncologianova GmbH, Recklinghausen, Germany, 5iOMEDICO, Freiburg, Germany, 6Praxis für interdisziplinäre Onkologie & Hämatologie, Freiburg, Germany 1

Introduction: Patients (pts) with non-small cell lung cancer (NSCLC) are mostly diagnosed with advanced disease and the majority receives chemotherapy. Both tumor burden and chemotherapy can strongly affect quality of life (QoL) but little is known about changes in QoL and other patient-reported outcomes (PROs) over time in pts in routine care. The LuLife project investigated QoL PROs during first line treatment in a real-world setting. Methods: LuLife was conducted as a satellite project of the tumor registry on lung cancer (TLK), which prospectively collected data on routine treatment of lung cancer patients from 256 office-based medical oncologists in Germany. It was started in 2010 and follow-up ended January 2016. In LuLife the validated questionnaires EORTC QLQ-C30 and -LC13 (lung cancer module) were sent to pts with NSCLC at start of 1st-line treatment and then every two months up to six times and they were filled in at home. Here, data on time to deterioration (TTD) of global QoL as well as cancer- and symptom-related PROs data are presented. TTD was investigated using the Kaplan-Meier-Method. Patients who reported a clinically meaningful change in a PRO at least once were defined as events. Results: 507 pts answered at least one questionnaire. At start of firstline treatment mean age was 66 years and 30%/56%/14% of pts had an ECOG status of 0/1/≥2, respectively. Questionnaires were answered by 485/424/353/290/240/194 pts at 0/8/16/24/32/40 weeks which represents a return rate of 96/87/81/76/73/65% of all pts alive at the respective time points. After 6 months 60% of pts had reported worsening of fatigue at least once; 50% of pts had reported worsening of pain, role, dyspnea, neuropathic symptoms, physical, cognitive and social functioning and 40% of pts had reported worsening of constipation, loss of appetite, insomnia, nausea and emotional functioning. Only 30% of pts reported mouth sore or diarrhea. Conclusion: Our data show that PROs of NSCLC pts deteriorate during the first month of treatment with fatigue, pain, neuropathy and dyspnea being the most frequently reported symptoms. Incorporation of PROs in routine care might assist physicians in addressing these symptoms. Disclosure: No conflict of interest disclosed. V987

Reptin drives tumor progression and chemotherapy resistance via HDAC1 recruitment in non-small cell lung cancer Mikesch J.-H.1, Schwammbach D.1, Hartmann W.1, Schmidt L.H.1, Schliemann C.1, Wiewrodt R.1, Marra A.2, Köhler G.3, Müller-Tidow C.4, Berdel W.E.1, Arteaga M.-F.1 University of Muenster, Faculty of Medicine, Muenster, Germany, 2Klinikum Bremen Ost, Bremen, Germany, 3Klinikum Fulda, Fulda, Germany, 4University of Halle/Saale, Faculty of Medicine, Halle, Germany 1

Introduction: While development of targeted NSCLC therapies has improved outcome for a subset of the patients, advanced and/or metastatic NSCLC remain a major therapeutic challenge. Thus, there is still high demand for identification of candidates for new-targeted therapies.

Abstracts

Methods: The role of reptin in adenocarcinoma and squamous cell carcinoma cells was assessed via shRNA and/or CRISPR Cas9 system mediated knock down in A549, HOP62 as well as LUDLU cells and subsequent functional in vitro and in vivo assays. Moreover, tissue array analyses were performed with primary patient samples of 307 NSCLC patients. Results: In this study, we identified the AAA+ ATPase reptin as a novel key player for NSCLC tumor progression. Reptin was upregulated in the large majority of NSCLC squamous cell and adenocarcinomas and conferred poor prognosis to the subset of patients with highly differentiated tumours. Knock down of Reptin in human NSCLC cells severely impaired their proliferation and self-renewal in vitro as well as in NSG xenograft mouse model in vivo. Moreover, we were able to show that Reptin directly interacts with histone deacetylase 1 (HDAC1) in NSCLC cells providing a potential new strategy for targeted therapies including HDAC inhibitors. In fact, selective HDAC1 inhibitor MS-275 substantially suppressed growth of human NSCLC cells and significantly sensitized them to treatment with cisplatin. Conclusion: Our findings provide important molecular insights into NSCLC oncogenesis and a novel avenue for targeted therapies. Disclosure: No conflict of interest disclosed. V988

Improved overall survival following implementation of nextgeneration sequencing in routine diagnostics of advanced lung cancer in Germany: results of the Network Genomic Medicine Kostenko A.1, Michels S.2, Kron F.1, Brandes V.2, Fischer R.2, Nogova L.2, Scheffler M.2, Fassunke J.3, Merkelbach-Bruse S.3, Scheel A.3, Ueckeroth F.3, Süptitz J.4, Gerigk M.4, Heydt C.3, Glossmann J.-P.1, Büttner R.3, Wolf J.2, for the Network Genomic Medicine Uniklinik Köln, Klinik I für Innere Medizin/ CIO Köln Bonn, Köln, Germany, Uniklinik Köln, Klinik I für Innere Medizin/ LCGC/ CIO Köln Bonn, Köln, Germany, 3 Uniklinik Köln, Institut für Pathologie/ CIO Köln Bonn, Köln, Germany, 4Uniklinik Köln, NGM/ CIO Köln Bonn, Köln, Germany 1 2

Introduction: Broad implementation of molecular diagnostics and personalized cancer care is hampered by insufficient molecular screening, missing reimbursement for comprehensive molecular testing and lack of access to appropriate drugs. The Network Genomic Medicine (NGM) Lung Cancer is a health care provider network offering comprehensive next generation sequencing (NGS)-based multiplex genotyping on a central diagnostics platform in Cologne for all inoperable lung cancer patients (pts) in Germany. Methods: The NGS panel used in NGM consists of 14 genes and 102 amplicons to cover potentially targetable aberrations. Mutation analyses were run on an Illumina (MySeq) platform, while FISH analyses were performed separately. In 2015, we have started the second outcome evaluation for all NGM pts who had received NGS-based molecular diagnostics. In particular, we have focused on molecular subgroups of EGFR, ALK, BRAF-V600E, HER2 and ROS1 positive pts and especially on NGM pts treated in clinical trials. Results: From 2013-2015 6210 lung cancer pts (n = 4244 non-squamous NSCLC) were genotyped. Preliminary data show the overall survival (OS) of 934 NSCLC pts including of 110 NSCLC pts treated in clinical trials. For 108 EGFR+ pts, the OS of clinical trials pts treated with so called 3rd generation EGFR-TKIs was 55 months (n = 25) vs 22 months in control group (n = 83) (p = 0,002; mean OS: 29 months; 95%CI: 36-83 months). For 85 ALK+ pts, the OS of pts treated in clinical trials was 35 months (n = 19) compared to OS of 23 months for 45 pts treated with one ALK inhibitor and 8 months for 19 pts treated with no ALK inhibitors (P < 0,0001; mean OS: 22 months; 95%CI: 22-33 months). Conclusions: While the first NGM evaluation in 2013 already showed a survival benefit of 2 years in EGFR-TKI treated EGFR+ pts compared to chemotherapy, our current evaluation in pts treated with 3rd generation EGFR-TKIs after acquired resistance to 1st gen. EGFR-TKIs shows the significant increasing of the OS. Similarly, we show a significant longer OS

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for ALK+ pts treated with 2 ALK inhibitors compared to treatment with one or no ALK inhibitor. Further results of this ongoing NGM evaluation will be provided.

V990

Disclosure: Anna Kostenko: No conflict of interest disclosed. Jürgen Wolf: Expert Testimony: Drittmittel für klinische Studien (IITs)

Subtype-specific activity in Liposarcoma (LPS) patients (pts) from a phase 3, open-label, randomized study of eribulin (ERI) versus dacarbazine (DTIC) in pts with advanced LPS and leiomyosarcoma (LMS)

V989

Chawla S.1, Schöffski P.2, Grignani G.3, Blay J.-Y.4, Maki R.G.5, D’Adamo D.R.6, Guo M.6, Demetri G.D.7

Pazopanib vs pazopanib / gemcitabine in refractory soft tissue sarcoma: The AIO-STS-009-trial Rüssel J.1, Lindner L.2, Reichard P.3, Heißner K.4, Kopp H.-G.5, Kessler T.6, Mayer-Steinacker R.7, Egerer G.8, Crysandt M.9, Kasper B.10, Niederwieser D.11, Kunitz A.12, Eigendorff E.13, Steighardt J.14, Cygon F.15, Meinert F.15, Stein A.16, Schmoll H.-J.15 Martin Luther University Halle-Wittenberg Halle, Department of Oncology and Hematology, Halle, Germany, 2University Hospital Großhadern, Ludwig Maximilians University, Department of Internal Medicine III, Munich, Germany, 3 HELIOS Klinikum Berlin-Buch, Berlin, Germany, 4University Hospital Tübingen, Department of Internal Medicine II, Tübingen, Germany, 5University Hospital Tübingen, Medical Center II, Tübingen, Germany, 6University Hospital Muenster, Medical Clinic A, Münster, Germany, 7University Hospital Ulm, Department for Internal Medicine III, Ulm, Germany, 8University Hospital Heidelberg, Department of Internal Medicine, Krankenhaus St. Vincentius, Heidelberg, Germany, 9University Hospital Aachen, Aachen, Germany, 10University Hospital Muenster, Münster, Germany, 11University Hospital Leipzig, Leipzig, Germany, 12Charité – Campus Virchow-Klinikum, Berlin, Germany, 13University Hospital Jena, Jena, Germany, 14Koordinationszentrum Klinische Studien, Halle, Germany, 15Martin Luther University, University Clinic, Halle, Germany, 16 Universitätsklinikum Hamburg Eppendorf, Hamburg, Germany 1

Background: This trial investigates the combination of pazopanib plus gemcitabine vs. pazopanib alone in anthracycline and /or ifosfamide pretreated or refractory advanced soft tissue sarcoma (STS), in a randomized multicenter phase II-trial associated with the soft-tissue-sarcoma study group of the AIO. Methods: Inclusion criteria: histo-/cytological confirmed STS, except Chondro-sa, Osteo-sa, PNET, GIST, mixed mesodermal uterus tumor; life expectancy > 3 months, ECOG-PS 0-2, adequate organ function, ≥ 1 measurable lesion (RECIST); relapse/progression after at least 1 prior line with anthracycline or ifosfamide or both. From 9/11 - 7/14, 90 patients have been randomized into Arm B (pazopanib) and Arm A (pazopanib plus gemcitabin). Primary endpoint (PE) is rate of progression @ 3 months (PFS-R), with an PFS-Rate of 40% (arm B) vs. =/> 60% (arm A), with 90 pts. Randomisation was stratified by liposarcoma (lipo-sa)-histology and center. Results: A total of 90 pts have been randomized (A: 42 / B: 44), and 86 pts are evaluable (ITT) (2 pts per arm not eligible). After a median follow up of 39 (21-54) months, the primary endpoint was met with 73,2% vs. 45,5% without progression after 3 months (HR 1.62 (CI: 1,16-2,27), p = 0,005) For all other histologies excluding lipo-sa the difference in PFS-rate was not significant (HR 1,24 (CI 0,89-1,74), p = 0,28), but in lipo-sa with 9/9 (100%, arm A) vs. 1/8 (12,5%, arm B) (HR 8 (CI 1,72-37,27), p < 0,001). Response-rates: (ITT): 12% (A) vs. 5% (B), w/o lipo-sa 15% vs. 6% (no response in lipo-sa in both arms, however SD 9/9 (A) vs. 2/8 (B)). PFS, adjusted for lipo-sa, was significantly improved with 5,6 (A) vs. 1,9 (B) months (median): HR 0,54 (CI 0,34-0,87), p = 0,01. For the lipo-sa subgroup the PFS-difference was striking with 8,6 (A) vs. 1,5 months (B), but not significant (p = 0,993). OS was not significant different for all pts, but a trend favouring the combination in lipo-sa (HR 0,42). Toxicity was different, as expected, but tolerabel, with mainly °3/4 hematological, cardiovascular, GI and renal; SAE N = 37 (A) vs 9 (B) with 2 fatal events in arm A (ARDS 1; unclear). Conclusion: Gemcitabin in addition to pazopanib significantly improves the PFS at 3 months and overall PFS, not OS, with strongest effect in liposa.

Sarcoma Oncology Center, Santa Monica, United States, 2University Hospitals Leuven, Leuven, Belgium, 3Fondazione del Piemonte per l’Oncologia IRCC, Candiolo, Italy, 4Université Claude Bernard & Centre Léon Bérard, Lyon, France, 5 Tisch Cancer Institute, Mount Sinai Medical Center, New York, United States, 6 Eisai Inc, Woodcliff Lake, United States, 7Sarcoma Center and Ludwig Center, Dana-Farber Cancer Institute and Harvard Medical School, Boston, United States 1

Background: A recent phase 3 study (NCT01327885, Schöffski et al. Lancet 2016) comparing ERI with DTIC in pts with advanced LPS or LMS, showed a significant improvement in overall survival (OS) for the ERI arm with a manageable toxicity profile. This analysis evaluated the efficacy and safety of ERI in LPS pts. Methods: Pts aged ≥18 yrs with advanced dedifferentiated, myxoid, round cell, or pleomorphic LPS incurable by multimodality therapy and from 3 geographic regions were included. Pts with ECOG status ≤2 and ≥2 prior systemic treatment regimens, including an anthracycline, were randomized 1:1 to ERI (1.4 mg/m2, IV on D1 and D8) or DTIC (850, 1000, or 1200 mg/m2, IV on D1) every 21-D until disease progression. OS, progression free survival (PFS), and safety were analyzed in LPS pts. Results: 143 pts with LPS (38% female; 76% < 65 yrs; 45% dedifferentiated, 39% myxoid/round cell, 16% pleomorphic LPS), representing 32% of the total study population, were included in this pre-planned analysis (71 ERI; 72 DTIC). Median OS for LPS pts receiving ERI vs DTIC was 15.6 vs 8.4 mo (HR = 0.51, [95% CI 0.35 0.75]; P = 0.001). OS benefit with ERI vs DTIC was observed independent of LPS histology (dedifferentiated—18.0 vs 8.1 mo, HR = 0.43 [95% CI 0.23, 0.79]; myxoid/round cell—13.5 vs. 9.6 mo, HR = 0.79 [95% CI 0.42, 1.49]; pleomorphic—22.2 vs 6.7 mo, HR = 0.18 [95% CI 0.04, 0.85]) and independent of geographic region. PFS in LPS pts for ERI vs DTIC was also improved (2.9 vs 1.7 mo, HR = 0.52, [95% CI 0.35-0.78]; P = 0.002). The mean number of treatment cycles for ERI vs DTIC was 6.5 and 3.2, respectively. In the ERI and DTIC arms, 100% and 96% of LPS pts had adverse events (AEs). Most frequent AEs in the ERI arm were alopecia (40%), fatigue (40%), neutropenia (39%), and nausea (39%). AEs of ≥grade (G) 3 occurred in 63% and 51% of LPS pts in the ERI and DTIC arms, respectively. Peripheral sensory neuropathy of G3 occurred in 3% of LPS pts in the ERI arm, with no incidence of G4 or G5. Conclusions: ERI was associated with a significant benefit in OS and PFS compared with DTIC in LPS pts and represents an active agent against LPS, sarcomas for which limited effective treatments are available. ClinicalTrials.gov identifier: NCT01327885 Funding Source: Eisai Inc. Disclosure: Sant Chawla: Advisory Role: Amgen, Threshold, CyDex, PharmaMar, Johnson & Johnson, Tracon, Morphotek; Financing of Scientific Research: Amgen, Threshold, CyDex, PharmaMar, Johnson & Johnson, Tracon, Morphotek; Expert Testimony: Amgen, Threshold, CyDex, PharmaMar, Johnson & Johnson, Tracon, Morphotek George Demetri: Employment or Leadership Position: Blueprint Medicines; Advisory Role: Bayer, Novartis, Pfizer, Lilly, EMD-Serono, Sanofi Oncology, Janssen Oncology, PharmaMar, Daiichi-Sankyo, Ariad, WIRB Copernicus Group, ZioPharm, Polaris Pharmaceuticals, Kymab, Genocea, Nektar Therapeutics, Blueprint Medicines, Kolltan Pharmaceuticals, G1 Therapeutics, Caris Life Sciences, Champions Oncology, Bessor Pharmaceuticals; Honoraria: Novartis; Expert Testimony: Bayer, Novartis, Pfizer, Janssen Oncology

Disclosure: No conflict of interest disclosed.

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Abstracts

CONTENTS AUTHOR INDEX

Freier Vortrag

Kopf-/Hals-Tumoren V995

Response to Lenvatinib treatment in patients with Radioiodine-refractory Differentiated Thyroid Cancer (RRDTC) Gianoukakis A.G.1, Mathias E.2, Dutcus C.E.2, Kalantari P.3, Yoon S.3 David Geffen School of Medicine, University of California Los Angeles, Los Angeles, United States, 2Eisai Inc, Woodcliff Lake, United States, 3Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center, Torrance, United States 1

Introduction: Lenvatinib (LEN) significantly prolonged median progression-free survival (PFS) vs placebo (PBO) in the primary analysis of the phase 3 SELECT study of RR-DTC (18.3 vs 3.6 mos; hazard ratio [HR] 0.21; 99% confidence interval [CI] 0.14-0.31; P < 0.001). Median duration of overall response (DOR; per RECIST 1.1) to LEN was not reached at the time of the primary analysis. Here we present new efficacy data with a focus on DOR. Methods: The data cutoff was 31 Aug 2015 (LEN: n = 261, PBO: n = 131); primary endpoint was PFS. DOR was examined for patients (pts) who had partial (PR) or complete responses (CR) and by subgroup (age, sex, tumor subtype, baseline disease burden, baseline Eastern Cooperative Oncology Group score, metastasis site, prior vascular endothelial growth factor [VEGF] therapy). For this analysis, tumor assessments were per investigator. We also report clinical features of 2 pts with RR-DTC with prolonged responses to LEN. Results: At cutoff, median PFS was 19.4 mos (95% CI 14.8-29.3) for LEN and 3.7 mos (95% CI 3.5-5.4) for PBO (HR 0.24; 99% CI 0.17-0.35; P < 0.0001). 157 Pts (60.2%) responded to LEN and median DOR was 30 mos (95% CI 18.4-35.2). Three pts (2.3%) responded to PBO and the median DOR was 14.7 mos (95% CI 7.5-not evaluable [NE; median not yet reached]). Median DOR to LEN was similar among subgroups except in pts with greater disease burden (tumor size ≤35mm: NE; 35-60mm: 27.5 mos; 60-92mm: 18.0 mos; >92mm: 15.7 mos), and pts with liver metastasis (yes: 15.7 mos; no: 31.3 mos). Two pts with prolonged DOR are still receiving LEN: 1 woman (Hürthle cell follicular thyroid cancer; no prior VEGF therapy) received LEN since Nov 2011 and achieved CR with a DOR of 46 mos (ongoing) and a PFS of 48 mos; 1 man (Hürthle cell variant of papillary thyroid carcinoma; no prior VEGF therapy) received LEN since Jul 2012 and achieved PR with a DOR of 35 mos (ongoing) and a PFS of 41 mos (of note, this pt has multiple metastases in the lung, abdomen, and bone). Conclusions: In this analysis, LEN PFS benefit was maintained along with a notable DOR. The case studies profiled demonstrate that an extended DOR to LEN can occur in pts with RR-DTC who show significant variation in the severity of disease. Disclosure: Andrew Gianoukakis: Advisory Role: Eisai Pharmaceuticals; Expert Testimony: Eisai, Roche, AstraZeneca Steve Yoon: No conflict of interest disclosed.

V996

Liquid biopsies reveal that acquired RAS-mediated resistance plays a major role in patients with head and neck cancer treated with cetuximab Voigtländer M.1, Braig F.1, Schieferdecker A.1, Busch C.-J.2, Laban S.2,3, Grob T.4, Kriegs M.5, Knecht R.2, Bokemeyer C.1, Binder M.1 Universitätsklinikum Hamburg-Eppendorf, Onkologisches Zentrum, II. Medizinische Klinik und Poliklinik, Hamburg, Germany, 2Universitätsklinikum Hamburg-Eppendorf, Klinik und Poliklinik für Hals-NasenOhrenheilkunde, Kopf-Hals-Chirurgie und -Onkologie, Hamburg, Germany, 3 Universitätsklinikum Ulm, Klinik für Hals-Nasen-Ohrenheilkunde, Ulm, Germany, 4Universitätsklinikum Hamburg-Eppendorf, Institut für Pathologie, Hamburg, Germany, 5Universitätsklinikum Hamburg-Eppendorf, Labor für Strahlenbiologie & Experimentelle Radioonkologie, Hamburg, Germany 1

Introduction: Resistance to epidermal growth factor receptor (EGFR) targeted therapy is insufficiently understood in head and neck squamous cell carcinoma (HNSCC), entailing the lack of predictive biomarkers that can be used for resistance monitoring. Patients and methods: We scanned the cetuximab epitope on the EGFR, KRAS, NRAS and HRAS for mutations by targeted next-generation sequencing (NGS) in 12 HNSCC cell lines and tumor samples from 46 cetuximab-treated HNSCC patients at baseline. In 20 patients of this cohort, we determined acquired mutations of EGFR and RAS genes by NGS of circulating tumor DNA (liquid biopsy) during and after cis- (or carbo-) platin/5-fluoruracil/cetuximab therapy. Results: None of the tumors and cell lines showed evidence of mutations in the cetuximab-interacting EGFR ectodomain or KRAS/NRAS at baseline, while activating HRAS mutations were found in two of 46 EGFR antibody-naïve patients (4.3%), well in line with previous reports. Liquid biopsy evaluation during cetuximab-based treatment revealed acquired HRAS, KRAS or NRAS mutations in about one third of patients. Six out of 13 patients (46%) with disease progression in the course of EGFR-directed treatment showed acquired RAS mutations, while no RAS mutations were found in the non-progressive subset of patients, indicating that the emergence of RAS mutant clones correlated significantly with clinical resistance (Chi square p = 0.032).The emergence of mutations preceded clinical progression in about half of the patients, with a maximum time from mutation detection to clinical progression of 16 weeks in our cohort. Conclusions: This study suggests for the first time that RAS mutations account for acquired resistance to EGFR targeting in a substantial proportion of HNSCC patients, even though these tumors are rarely mutated at baseline. Liquid biopsy monitoring may help to detect resistant subclones before overt clinical resistance occurs. Disclosure: No conflict of interest disclosed. V997

Evaluation of systemic inflammatory response (SIR) markers in recurrent or metastatic head and neck squamous cell carcinoma (r/m HNSCC) Pogorzelski M.1, Hilser T.1, Ting S.2, Vossebein I.1, Gauler T.C.3, Abendroth A.1, Lang S.4, Stuck B.4, Abu-Jawad J.3, Stuschke M.3, Schmid K.W.2, Schuler M.1, Kasper S.1 Innere Klinik (Tumorforschung), Universitätsklinikum Essen, Essen, Germany, Institut für Pathologie, Universitätsklinikum Essen, Essen, Germany, 3Klinik für Strahlentherapie, Universitätsklinikum Essen, Essen, Germany, 4Klinik für HalsNasen-Ohrenheilkunde, Universitätsklinikum Essen, Essen, Germany 1 2

Introduction: Intratumoral inflammation is a known hallmark of cancer. In r/m HNSCC platinum based immuno-chemotherapy with the anti-EGFR antibody cetuximab remains the standard of care. Recently the immune checkpoint inhibitor nivolumab demonstrated superior outcome after failure of first line therapy in a phase III trial. However, predictive biomarkers for the response to these immunotherapies are missing. Against this background we evaluated the prognostic and predictive value of the systemic inflammatory response (SIR) markers: modified Glasgow Prog-

Abstracts

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nostic Score (mGPS), CRP, anemia, Platelet-/Lymphocyte Ratio (PLR), Neutrophile-/Lymphocyte Ratio (NLR) and Lymphocyte-Monocyte Ratio (LMR) in patients with r/m HNSCC treated with immuno-chemotherapy at the West German Cancer Center (WTZ). Methods: SIR markers were retrospectively evaluated in 105 patients (pts) with r/m HNSCC at time point of start of palliative systemic treatment. Cutoff values for PLR were 357, NLR were 8.3 and LMR were 2.2 according to the median values for the entire cohort. For CRP, values above 10mg/l were selected and anemia was defined below 13g/dl. The mGPS is based on CRP and albumin. SIR markers were correlated with clinic-pathological findings, treatment response, progression free (PFS) and overall survival (OS) using Kaplan-Meier curves and Cox proportional hazard models. Results: Median OS was 12.6 months (95% CI 10.1-15.1). High CRP, anemia and mGPS>0 were significantly associated with reduced OS. PLR was significantly associated with reduced OS and PFS upon cetuximab based therapy (table 1). The median OS for pts with high PLR was 7.5 vs 13.9 months (HR 1.716), for anemia: 9.2 vs 24.1 months (HR 2.739), for high CRP: 9.2 vs 17.1 months (HR 2.029) and for mGPS>0: 10.6 vs 27.0 months (HR 2.752). NLR and LMR had no impact on OS or PFS. Conclusion: SIR biomarkers, routinely assessed in clinical settings can be easily used as prognostic markers to optimize palliative systemic treatment. To overcome the negative prognostic impact of SIR markers tailored treatment strategies for these subgroups should be evaluated in clinical trials.

Methods: This was a single-arm, open-label, phase 2 study conducted in Japan with an updated data cutoff of 9 Jul 2015. Pts received LEN 24 mg/d in 28-d cycles until progressive disease or development of unacceptable toxicity. Primary endpoint was safety; secondary endpoint was efficacy as assessed by PFS, overall survival (OS), overall response rate (ORR), and disease control rate (DCR). Results: 51 Pts, including 25 pts with RR-DTC (previously 23), 9 pts with MTC, and 17 pts with ATC (previously 11) were enrolled. All pts had at least 1 treatment-emergent adverse event (TEAE). The most common any-grade TEAEs included hypertension (90%), decreased appetite (78%), palmar-plantar erythrodysaesthesia syndrome (77%), fatigue (73%), proteinuria (61%), stomatitis (57%), and diarrhea (55%). Incidences of Grade 3 and 4 TEAEs were similar among subgroups (RR-DTC, 72%; MTC, 100%; ATC, 88%). Of note, only 1 pt discontinued treatment due to a TEAE. There were 4 fatal serious AEs, all considered unrelated to LEN. Tumor shrinkage occurred in most pts, including pts with ATC. Median duration of treatment was 5.5 mos (range, 0.7-33.1) for pts with ATC; 8 received LEN for more than 6 mos. Efficacy parameters are shown in the table below. Conclusions: In this study, LEN showed tumor shrinkage in almost all pts with advanced thyroid cancer, including pts with ATC. Toxicities were manageable with dose modifications. LEN efficacy, especially in ATC, warrants further investigation. Tab. 1.

RR-DTC (n = 25) Median PFS 25.8 (18.4-NE) (95% CI)—mos. Median OS 31.8 (31.8-NE) (95% CI)—mos. ORR—n (%)* 17 (68) DCR—n (%) 25 (100) NE, not evaluable. *All partial responses.

Tab. 1. SIR marker and OS in m/r HNSCC

SIR marker

OS since r/m (months)

p-value

PLR>357 vs <357

7.5 vs 13.9

0.021

NLR>8.3 vs <8.3

7.5 vs. 12.8

0.453

LMR<2.2 vs >2.2

9.2 vs. 16.8

0.167

CRP > 10mg/l vs <10mg/l

9.2 vs. 17.1

0.003

mGPS>0 vs 0

10.6 vs. 27.0

0.02

Anemia<13g/dl vs >13g/dl

9.2 vs. 24.1

<0.001

HR (95% CI), p-value 1.716 (1.0772.734), 0.023 1.213 (0.7312.015), 0.454 1.425 (0.8592.364), 0.170 2.029 (1.2563.279), 0.004 2.752 (1.1426.635), 0.024 2.739 (1.6654.506), <0.001

V998

Takahashi S. , Kiyota N. , Yamazaki T. , Chayahara N. , Nakano K. , Inagaki L.1, Toda K.4, Enokida T.3, Minami H.2, Imamura Y.2, Sasaki T.5, Suzuki T.5, Fujino K.5, Dutcus C.6, Tahara M.3 3

Department of Medical Oncology, Cancer Institute Hospital of JFCR, Tokyo, Japan, 2Department of Medical Oncology and Hematology, Kobe University Hospital, Kobe, Japan, 3Department of Head and Neck Medical Oncology, National Cancer Center Hospital East, Kashiwa, Japan, 4Department of Head and Neck Oncology, Cancer Institute Hospital of JFCR, Tokyo, Japan, 5Eisai Co., Ltd., Tokyo, Japan, 6Eisai Inc., Woodcliff Lake, United States 1

Background: Thyroid cancer is classified into differentiated (DTC), medullary (MTC), and anaplastic (ATC) types. Lenvatinib (LEN) significantly prolonged progression-free survival (PFS) vs placebo in the phase 3 SELECT trial of patients (pts) with 131I-refractory DTC (RR-DTC). Initial results of this phase 2 trial (Study 208) of LEN in RR-DTC, MTC, and ATC were reported with a cutoff date of 15 Jun 2014. Here we report the final results of this study.

308

9.2 (1.8-NE)

7.4 (1.7-12.9)

12.1 (3.8-NE)

10.6 (3.8-19.8)

2 (22) 9 (100)

4 (24) 12 (71)

Disclosure: Shunji Takahashi: Financing of Scientific Research: Eisai; Expert Testimony: Eisai Makoto Tahara: Advisory Role: Merck, Serono, Bristol Myers-Squibb; Financing of Scientific Research: Merck, Serono, Bristol Myers- Squibb; Expert Testimony: Eisai, Boehinger Ingelheim

What can a qualified medical oncological and especially palliative care do for patients with head and neck cancer? - a retrospective analysis of 156 cases

Phase 2 study of Lenvatinib in patients with differentiated, medullary, and anaplastic thyroid cancer: Final analysis results 2

ATC (n = 17)

V999

Disclosure: No conflict of interest disclosed.

MTC (n = 9)

Oncol Res Treat 2016;39(suppl 3):1–327

Große-Thie C.1, Kriesen U.1, Kragl B.1, Neuer A.1, Freitag S.1, Leithäuser M.2, Gläser D.1,3, Hildebrandt G.4, Punke C.5, Frerich B.6, Junghanss C.1 Department of Medicine, Clinic III – Hematology/Oncology/Palliative Care, Rostock University Medical Center, Rostock, Germany, 2Hämatologische Schwerpunktpraxis, Rostock, Germany, 3Clinic of Internal Medicine III – Hematology/Oncology/Hemostaseology, Palliative Care, Complementary Medicine, Klinikum Südstadt Rostock, Rostock, Germany, 4Department of Radiotherapy, Rostock University Medical Center, Rostock, Germany, 5 Department of Oto-Rhino-Laryngology, Head & Neck Surgery, Rostock University Medical Center, Rostock, Germany, 6Department of CranioMaxillofacial-Surgery, Rostock University Medical Center, Rostock, Germany 1

Introduction: The comprehensive treatment of head and neck (h&n) cancer patients (pts) requires a multidisciplinary team consisting of surgeons, radiation and medical oncologists. Usually the later guide treatment in neoadjuvant concepts and locally advanced or metastatic disease. Advanced h&n cancer pts are frequently characterised by high symptom burden requiring specialized care including palliative care (PC). In general management is challenging and some issues such as sought place of dying might be difficult to address. Here we report on a large cohort of h&n cancer pts and characterize them in regards to PC issues.

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CONTENTS AUTHOR INDEX

Methods: Consecutive h&n cancer pts that were seen in our medical oncology as well as PC department between 1/2010 and 12/2015 were included. Parameters analysed included age, sex, preceded treatments, employed chemotherapeutic protocols and duration of contact, survival and place of death Results: The majority of the pts were male (89%). The median age at point of first contact was 60 (32-98) years. Most pts had received curative intended treatments earlier: radiation ± chemotherapy (77%) or surgery (79%). Only 16 pts (10%) received neoadjuvant treatment, mostly within prospective study concepts e.g. TISOC. A total of 143 pts had advanced metastatic disease, of which 58 pts (41%) received palliative immunochemotherapy, 19 patients (13%) single immunotherapy with cetuximab and 66 patients (46%) a best supportive care concept. In case of palliative chemotherapeutic care 63% received at least one therapy line, 27% two lines and only 10% of patients >2 lines. 22 patients (14%) are currently alive, in 126 cases (80%) death was confirmed while 8 (6%) were lost to follow up. About 75% of the patients (117/156) received special palliative care (palliative care ward or special ambulant palliative care concept). Place of dying was evaluable for 100 pts. Only 5 patients (5%) died at home, 57 (57%) in the palliative care unit, 31 (31%) in other hospitals and 7 (7%) in the oncological care unit. Conclusion: First, dying at home is a rare event in h&n cancer pts. Second, palliative care approaches are often implemented in advanced disease pts leading to a frequent dying within a palliative care unit. Finally, for optimal oncological treatment the early implementation of PC seems to be advisable in order to optimize symptom burden control and increase therapy compliance. Disclosure: No conflict of interest disclosed. V1000

Targeted next-generation sequencing identifies molecular subgroups in squamous cell carcinoma of the head and neck with distinct outcome after concurrent chemoradiation Tinhofer I.1, Stenzinger A.2, Eder T.1, Konschak R.1, Niehr F.1, Endris V.2, Distel L.3, Hautmann M.G.4, Mandic R.5, Stromberger C.1, Weichert W.6, Budach V.1 Charité Universitätsmedizin Berlin, Klinik für Radioonkologie und Strahlentherapie, Berlin, Germany, 2Universitätsklinikum Heidelberg, Institut für Pathologie, Heidelberg, Germany, 3Universitätsklinikum Erlangen, FriedrichAlexander Universität Erlangen-Nürnberg, Klinik für Radioonkologie und Strahlentherapie, Erlangen, Germany, 4Universitäsklinikum Regensburg, Klinik für Radioonkologie und Strahlentherapie, Regensburg, Germany, 5 Universitätsklinikum Giessen-Marburg, Marburg, Germany, 6Technische Universität München, Institut für Pathologie, München, Germany 1

Background: Based on epidemiological (HPV status, smoking habits) and clinical risk factors (T/N stage) three subgroups of patients suffering from locally advanced oropharyngeal carcinoma with significantly different outcome after concurrent chemoradiation (cCRTX) can be distinguished. Mutational profiling by targeted next-generation sequencing (NGS) might further improve risk stratification. Patients and methods: Patients with stage IV squamous cell carcinoma of the oropharynx and hypopharynx who had been enrolled in a randomized phase III trial (ARO-0401) comparing two regimens of cCRTX and from whom archival tumor specimens were available were included in this study. The HPV status was determined by p16 immunostaining and detection of HPV DNA. Targeted NGS covering 45 genes frequently altered in squamous cell carcinoma of the head and neck (SCCHN) was applied for detection of non-synonymous somatic and germline mutations. Mutational profiles of risk groups and their interference with the efficacy of cCRTX were determined. Results: The prognostic value of the ‘Ang’ risk model could be confirmed in the total biomarker study cohort (N = 175) as well as the patient subgroup for which mutational profiles could be established (N = 97). Mutations in genes involved in phosphoinositide 3-kinase (PI3K), receptor tyrosine kinase (RTK) and p53 signaling pathways were significantly enriched in the low- (N = 7), intermediate- (N = 20), and high-risk group

Abstracts

(N = 70), respectively. Mutations in TP53 identified a subgroup of highrisk patients with dismal outcome after cCRTX whereas no prognostic relevance was observed for mutations in PI3K and RTK signaling pathways in the low- and intermediate risk groups, respectively. Mutated NOTCH1 and two functional KDR germline variants (rs2305948, rs1870377) were associated with improved outcome in all risk groups. All genetic markers (TP53, NOTCH1, KDR) remained independent prognosticators of OS in the multivariate model. Conclusion: A potential of targeted NGS for risk classification of SCCHN cases beyond HPV status and clinical factors was demonstrated. Disclosure: Ingeborg Tinhofer: Expert Testimony: Merck, Pfizer, Genentech Volker Budach: Financing of Scientific Research: Merck, Sanofi, Amgen, Accuray; Expert Testimony: Merck

Fortbildung

Kompetenznetz Maligne Lymphome V1005

Aggressive lymphomas Pfreundschuh M.1, Deutsche Studiengruppe für Hochmaligne NonHodgkin-Lymphome Klinik für Innere Medizin, Medizin I, Homburg, Germany

Young patients with no risk factor according to the age-adjusted (aa) IPI and no bulky disease have a high event-free and nearly 100% overall survival after 6 cycles of R-CHOP-21, suggesting that some of these patients are overtreated. For the first time in the history of DLBCL treatment prospective trials like the FLYER study evaluate reduction of treatment from 6 to only 4 cycles of CHOP with 6 applications of rituximab. For young patients with aaIPI = 1 and/or bulky disease, best results have been achieved with 6xR-CHOP-21 plus involved-field radiotherapy to bulky disease or the more aggressive and more toxic R-ACVBP program without radiotherapy. For young poor-prognosis patients (aaIPI = 2,3) best results have been achieved with 8xR-CHOEP-14, with a 3-year overall survival of aaIPI = 2 patients of roughly 90%, demonstrating that this subpopulation is no high-risk population any more in the rituximab era. For young aaIPI = 3 patients there is still room for improvement and ongoing trials evaluate primary high-dose chemotherapy with stem cell transplantation or the addition of new drugs to a CHOP(E)P-14 backbone. Elderly patients who represent up to two thirds of all DLBCL patients do best with 8 cycles of R-CHOP-21 or 6 cycles of R-CHOP-14 plus 2 additional rituximab administrations. Since further intensification of chemotherapy is hardly possible in the elderly population, improvement strategies evaluate new doses and schedules of rituximab based on the recent demonstration that rituximab is suboptimally dosed in all DLBCL patients except elderly females. Other improvement strategies include vitamin D substitution in vitamin D insufficient and deficient DLBCL patients, because vitamin D deficiency impairs ADCC, the most important mechanism of action of rituximab, as well as new drugs targeting specific molecules involved in the signal transduction pathway of the B-cell receptor. There are several novel concepts in clinical trials, the most promising being BTK and BCL2 inhibitors, with BARs (B-cell receptor antigens for reverse targeting) providing an ultimate specificity restricted to the malignant B-cell clone. Finally, checkpoint inhibitors that have proven efficacious in many types of malignancies also hold promise for the treatment of malignant lymphomas. While the rate of patients dying from DLBCL has been cut into half during the last decade, these new approaches should further improve the cure rate of DLBCL in the near future. Disclosure: Michael Pfreundschuh: Other Financial Relationships: Advisory Boards Celgene, Roche, Spectrum, Takeda

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V1007

V1008

Phase 2a study of the phosphatidylinositol-3-kinase (PI3K) inhibitor copanlisib in patients with relapsed/refractory, indolent or aggressive lymphoma

Multiple Myeloma

Dreyling M.H.1, Morschhauser F.2, Bouabdallah K.3, Cunningham D.4, Bron D.5, Linton K.6, Assouline S.7, Verhoef G.8, Thieblemont C.9, Vitolo U.10, Garcia-Vargas J.11, Gorbatchevsky I.11, Neves M.12, Grunert J.13, Hiemeyer F.14, Childs B.H.11, Zinzani P.L.15 Universitätsklinik Grosshadern/LMU, Medizinische Klinik III, München, Germany, 2Franck Morschhauser, Hematology Department, Lille, France, 3 University Hospital of Bordeaux, Service d’Hématologie et de Thérapie Cellulaire, Bourdeaux, France, 4The Royal Marsden Hospital, GI & Lymphoma Unit Department of Medicine, Sutton, United Kingdom, 5Institute Jules Bordet, (ULB), Brussels, Belgium, 6The Christie NHS Foundation Trust, Department of Haemato-oncology, Manchester, United Kingdom, 7Jewish General Hospital, McGill University, Hematology, Montreal, Canada, 8University Hospital Leuven, Leuven, Belgium, 9APHP-Hôpital Saint-Louis, Department of Hemato-oncology, Paris, France, 10Città della Salute e della Scienza di Torino, Department of Oncology and Hematology, Torino, Italy, 11Bayer HealthCare Pharmaceuticals, Whippany, United States, 12Bayer HealthCare, Sao Paulo, Brazil, 13Bayer Pharma AG, Wuppertal, Germany, 14Bayer Pharma AG, Berlin, Germany, 15University of Bologna, Institute of Hematology ‘Seragnoli’, Bologna, Italy 1

Introduction: Copanlisib (BAY 80-6946) is a selective pan-class I PI3K inhibitor with potent activity against the δ- and α-isoforms. We report here on the efficacy and safety of copanlisib in subjects with relapsed/refractory indolent or aggressive non-Hodgkin lymphoma (NHL) or chronic lymphocytic leukemia (CLL). (NCT01660451) Methods: Patients with histologically confirmed indolent or aggressive lymphoma or CLL, relapsed or refractory to ≥2 prior lines of treatment were enrolled. Copanlisib (0.8 mg/kg) was administered intravenously on days 1, 8 and 15 of a 28-day cycle until disease progression or unacceptable toxicity. The primary endpoint was objective response rate (ORR) as assessed by independent radiology review according to the response criteria for lymphoma (Cheson et al, JCO 1999) or the guidelines for diagnosis and treatment of CLL (Hallek et al, Blood 2008). Secondary endpoints included progression-free survival (PFS) and duration of response (DOR), safety and tolerability. Results: A total of 20 patients with indolent NHL (iNHL; 16 FL, 3 marginal zone lymphoma, 1 small lymphocytic lymphoma) or CLL (n = 13) and 48 with aggressive NHL [15 DLBCL, 14 peripheral T-cell lymphoma, 11 mantle cell lymphoma (MCL), 6 transformed, 1 mediastinal, and 1 Grade 3b FL] were treated; median age 67 years; 53% male; median 3 (range 1-10) number of previous lines of treatment and prior rituximab in 80% of patients. At the time of analysis, the ORR was 47% (90% CI 27-68) and stable disease (SD) was 47% (CI 27-68) in patients with iNHL, 38% (CI 17-65) and 46% (CI 22-71) in CLL, and 26% (CI 16-39) and 17% (CI 9-29) in aggressive NHL, respectively. For patients with FL, the ORR was 40% (CI 19-64), with 1 CR, 2 uCRs, and 3 PRs; SD in 53% (CI 30-76). The ORR in the MCL patients was 64% (CI 35-86; 2 uCR and 5 PRs). The median DOR was 390 days in the indolent group and 166 days in the aggressive group. The most common treatment-related adverse events (AEs) of all grades (G) were hyperglycemia (59%), hypertension (54%), diarrhea (33%), and fatigue (28%). G3-4 treatment-related AEs occurring in >10% of patients included hypertension (39%) and hyperglycemia (25%). There were 4 treatment-related grade 5 events: meningitis, respiratory failure and 2 lung infections. Conclusions: Copanlisib has promising activity with a manageable toxicity profile as treatment for patients with relapsed/refractory indolent or aggressive lymphoma. Disclosure: No conflict of interest disclosed.

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Einsele H.1 University Hospital Würzburg, Internal Medicine II, Würzburg, Germany

The criteria for initiating the antiproliferative therapy for patients with Multiple Myeloma have been changed recently. According to the international guidelines a patient with documented Multiple Myeloma should be treated even if no CRAB criteria can be documented in case the bone marrow infiltration is > 60%, the light chain ratio is > 100 or < 0,01 or there is more than 1 focal lesion on the MRI. Argument for these new criteria is that in the majority of patients with at least one of these three new criteria there is a progression to symptomatic myeloma and end organ disease in less than two years. For the treatment of younger patients with Multiple Myeloma both German study groups still offer stem cell transplantation up to the age of 70 or even 75 years. Currently the DSMM XIII study for elderly patients (patients ≥ 65 years) evaluates whether in addition to the established Rd therapy and according to the FIRST Study an additional high dose chemotherapy and autologous stem cell transplantation increases the rate of complete remissions and the progression free and overall survival. For the younger patients from both German study groups (DSMM and GMMG) evaluate new induction therapies. In both study groups two novel agents are tested in the induction protocol and in addition the role of monoclonal antibodies in the induction treatment is evaluated. Another important question is whether patients that are benefitting from the first transplantation by achieving at least a very good partial remission benefit from an additional high dose chemotherapy and autologous stem cell transplantation. In addition the role of allogeneic stem cell transplantation is studied for patients with a suboptimal response after the first autograft and furthermore the role of antibody constructs in consolidation and maintenance is studied after transplantation. Disclosure: Hermann Einsele: Advisory Role: Janssen, Amgen, Novartis, Celgene; Financing of Scientific Research: Janssen, Amgen, Novartis, Celgene; Expert Testimony: Janssen, Amgen, Novartis, Celgene

Freier Vortrag

Allogene Stammzelltransplantation V1010

High pre-transplant serum asymmetric dimethylarginine (ADMA) levels are associated with worse outcome after allogeneic stem cell transplantation Radujkovic A.1, Dietrich S.1, Ho A.D.1, Dreger P.1, Luft T.1 Universitätsklinikum Heidelberg, Innere Medizin V, Heidelberg, Germany

Introduction: Graft-versus-host disease (GVHD) is the major complication contributing to non-relapse mortality (NRM) after allogeneic stem cell transplantation (alloSCT). Mortality after acute GVHD is associated with endothelial cell distress. In previous studies, we and others could show that endothelial cell associated markers that predict NRM and NRM after GVHD can be defined already prior to alloSCT. Asymmetric dimethylarginine (ADMA) is an endogenous nitric-oxide-synthase inhibitor causing endothelial cell dysfunction. In this retrospective study, we investigated the impact of pre-transplant serum ADMA levels on outcome parameters in 504 patients allografted at our institution. Methods: Determination of serum ADMA levels was performed using the ADMA Xpress ELISA Kit® (Immundiagnostik AG, Bensheim, Germany) according to the manufacturer’s instructions. Results: Median pre-transplant ADMA level was 0.69 (range 0.182.0) µM. In univariable analyses and as continuous variable, increasing pre-transplant ADMA levels were associated with worse OS (HR 1.57, P = 0.015) and a trend toward higher NRM incidence (HR 1.69, P = 0.06) but not relapse (HR 1.06, P = 0.82). Although not significantly associated

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with GVHD (P = 0.09), increasing pre-transplant ADMA levels were associated with increased incidence of NRM after GVHD onset (HR 2.81, P = 0.05) but not with relapse (HR 0.82, P = 0.81) translating into significantly worse OS after GVHD onset (HR 2.90, P = 0.01). In multivariable analyses adjusting for known confounders, increasing pre-transplant ADMA levels emerged as an independent predictor of worse survival (HR 1.62, P = 0.01) and higher incidence of NRM (HR 1.64, P = 0.07) without influencing relapse rates (HR 1.14, P = 0.58). When an ADMA cut-off of 1.45 µM (derived after maximally selected rank statistics) was applied to stratify patients in high (n = 35) and low (n = 469) ADMA groups, patients with high pre-transplant ADMA levels had increased NRM incidence (HR 2.20, P = 0.03) which translated into inferior survival (HR 2.70, P < 0.001). Conclusions: Patients at high risk of death can be identified already prior to alloSCT by elevated serum levels of ADMA, a relevant marker of endothelial cell physiology and established risk factor for (cardio)vascular disease. Further studies on the role of the endothelium and markers associated with endothelial distress in the setting of alloSCT are highly warranted. Disclosure: No conflict of interest disclosed.

Our results imply an association between HLA-E MM and better HSCT outcome in AL patients. A putatively enhanced NK alloreactivity due to HLA-E MM may account for this observation, as it has been demonstrated that NK alloreactivity associates with lower GvHD rates. In vitro studies could help unravel the biological mechanisms implicated. In conclusion, our study is the first to provide data on the effect of HLA-E matching in an AL MUT context. Confirmational, larger cohort studies are yet warranted before further conclusions can be drawn. This work was supported by the Deutsche José Carreras Leukämie-Stiftung e.V. (Grant No. DJCLS 11/10) Disclosure: No conflict of interest disclosed. V1012

The prognostic impact of differential GLI1 expression in patients with acute myeloid leukemia after non-myeloablative allogeneneic hematopoietic stem cell transplantation Bill M.1, Jentzsch M.1, Schuhmann L.1, Grimm J.1, Knyrim M.1, Schmalbrock L.1, Schubert K.1, Cross M.1, Vucinic V.1, Franke G.-N.1, Pönisch W.1, Behre G.1, Lange T.1, Niederwieser D.1, Schwind S.1 Universität Leipzig, Hämatologie und Internistische Onkologie, Leipzig, Germany 1

V1011

HLA-E mismatch improves HSCT outcome in acute leukemia patients Tsamadou C.1,2, Fürst D.1,2, Niederwieser D.3, Bunjes D.4, Neuchel C.1,2, Gramatzki M.5, Arnold R.6, Wagner E.7, Einsele H.8, Schrezenmeier H.1,2, Mytilineos J.1,2,9 Institute of Clinical Transfusion Medicine and Immunogenetics Ulm, Transplantation Immunology, Ulm, Germany, 2Institute of Transfusion Medicine, University of Ulm, Ulm, Germany, 3University of Leipzig, Department of Hematology/Oncology, Leipzig, Germany, 4University of Ulm, Department of Internal Medicine III, Ulm, Germany, 52nd Department of Medicine, University of Kiel, Division of Stem Cell Transplantion and Immunotherapy, Kiel, Germany, 6 Charité Campus Virchow Berlin, Hematology/Oncology Department, Berlin, Germany, 7Johannes Gutenberg-University, Department of Internal Medicine III, Mainz, Germany, 8University Hospital Würzburg, Department of Internal Medicine II, Würzburg, Germany, 9DRST – German Registry for Stem Cell Transplantation, Ulm, Germany 1

While the immunomodulatory properties of HLA-E have long been described, its role in hematopoietic stem cell transplantation (HSCT) remains markedly elusive. The aim of this study was to investigate the impact of HLA-E as alloreaction-mediator in an acute leukemia (AL) matched unrelated HSCT (MUT) setting. 552 AL (AML:329, ALL:151, AL:72) patients (P) undergone 10/10 HLA MUT and their donors (D) were HLA-E genotyped by sequence based typing. Three alleles (01:01, 01:03, 01:07) and four genotypes (01:01/01:01, 01:01/01:03, 01:03/01:03, and 01:01/01:07) were identified. The effect of P and D HLA-E genotype as well as of P-D HLA-E match grade were assessed using univariate Kaplan-Meier (KM), multivariate Cox regression, and competing risks analyses. OS (overall survival), DFS (disease free survival), RI (relapse incidence) and NRM (non-relapse mortality) were set as endpoints, while statistical significance was set to a p ≤ 0.05. The HLA-E allele frequencies found for both, P and D, accorded with those in literature for Caucasian populations. 349 of 552 (63%) pairs were HLA-E matched (M) and 203 (37%) were HLA-E mismatched (MM) with balanced distribution of patients in each group with respect to all important clinical predictors. KM analysis revealed notably improved 5y OS (54% vs 39%, p = 0.001) and DFS (45% vs 33%, p = 0.008) in the HLA-E MM compared to the HLA-E M cases. Multivariate analysis confirmed these results for both, OS (HR = 0.66, CI = 0.50-0.87, p = 0.003) and DFS (HR = 0.73, CI = 0.57-0.94, p = 0.013). NRM was also significantly lower in the MM group as shown by both, univariate (24% vs 34%, p = 0.008) and multivariate (HR = 0.64, CI = 0.44–0.92, p = 0.017) analyses. Interestingly, the “HLA-E mismatch” effect was far more intense among advanced disease P. Neither P, nor D HLA-E genotype had any clear impact on the study endpoints.

Abstracts

Introduction: Hedgehog signaling is crucial in embryonic development, stem cell biology & leads to activation of GLI1. In mice, loss of Gli1 increased the number of quiescent long-term hematopoietic stem cells with enhanced engraftment capacities & impaired myeloid development. The prognostic impact of GLI1 expression in human AML is inconclusive. In non-myeloablative conditioning (NMA) followed by hematopoietic stem cell transplantation (HCT) the therapeutic approach bases on an immunological graft-versus-leukemia (GvL) effect. Here, we tested the association of GLI1 expression with outcome in AML pts undergoing NMA-HCT. Methods: We analyzed 135 pts (median age 64 years (y), range 38–75y) who received NMA-HCT (fludarabine & 2Gy total body irradiation) at our institution between 01/2000 & 06/2012 with pretreatment bone marrow (BM) available. Median follow-up was 4.3y. A myeloid standard surface marker panel was assessed. FLT3-ITD, FLT3TKD & expression status of EVI1, BAALC, ERG, MN1, miR-9 & miR-181a as well as the mutation status of NPM1, CEBPA, IDH1, IDH2 & DNMT3A genes were determined. GLI1 expression was measured by qPCR & normalized to 18S. The third quartile cut defined high expressers. Results: Pts with high GLI1 expression had higher % of blasts in BM (p < 0.01) & peripheral blood by trend (p = 0.10), had higher % of CD14 (p = 0.05) & lower % of CD45 (p = 0.01) positive cells, were less likely to have a monosomal karyotype by trend (p = 0.07) & to express EVI1 (p = 0.05) & more likely to have a IDH2 mutations (p = 0.01). High GLI1 associated with significantly increased MN1 (p < 0.01), BAALC (p = 0.02) & ERG (p < 0.01) expression & lower miR-9 (p = 0.01) & miR-181a (p < 0.01) expression. High GLI1 expressers had a significantly longer overall survival (OS; p < 0.01) & event-free survival (EFS; p = 0.10; Figure 1) by trend. In multivariable analysis, high GLI1 expression associated with longer OS (Hazard Ratio 0.42; 95% Confidence Interval 0.23-0.79; p < 0.01). Conclusion: A high GLI1 expression independently impacted on outcome in AML pts receiving NMA-HCT. Further mechanistic studies are needed to explore these results biologically & clinical studies are necessary to confirm our findings.

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Fig. 1b. GPR56.

Fig. 1a. Disclosure: No conflict of interest disclosed. V1013

High expression of the stem cell marker GPR56 associates with an increased relapse incidence in Acute Myeloid Leukemia (AML) patients (pts) undergoing allogeneic stem cell transplantation (HSCT) Jentzsch M.1, Bill M.1, Schumann L.1, Grimm J.1, Schulz J.1, Knyrim M.1, Franke G.-N.1, Behre G.1, Pönisch W.1, Vucinic V.1, Müller-Tidow C.2, Pabst C.2, Niederwieser D.1, Schwind S.1 Universitätsklinik Leipzig, Hämatologie und internistische Onkologie, Leipzig, Germany, 2Universitätsklinik Halle (Saale), Hämatologie und Onkologie, Halle (Saale), Germany 1

Leukemia initiating cells (LIC) are thought responsible for AML propagation. The recently described G protein-coupled receptor 56 (GPR56) was shown to define a novel LIC compartment independently of the established CD34+/CD38- phenotype & to contribute to disease aggressiveness. HSCT as consolidation therapy bases on immunological graft-versus-leukemia (GvL) effects. The prognostic impact of aberrant GPR56 expression in the context of the CD34+/CD38- burden at diagnosis in AML pts receiving HSCT is unknown. We analyzed 148 AML pts (age 61, range 14–75 years [y]) receiving non-myeloablative (78%, Fludarabine + 2Gy total body irradiation [TBI]) or myeloablative (22%, Cyclophosphamide + 12Gy TBI) HSCT in complete remission (CR, 88%) or CR with incomplete peripheral recovery (12%). We assessed the mutation (mut) status of CEBPA, NPM1, RUNX1 & FLT3-ITD & BAALC & EVI1 expression at diagnosis. Flow cytometry determined the CD34+/CD38- burden & a 5% cut-off defined pts with a high & low CD34+/CD38- burden at diagnosis. GPR56 was measured by qRT-PCR, normalized to ABL1 & a median cut defined high & low expressers. Median follow up was 4.9y. At diagnosis high GPR56 expressers had lower white blood counts (P = 0.04), were less likely to have de novo AML (P = 0.02), core-binding factor AML (P = 0.01), a NPM1mut (P = 0.02) & more likely to have del7 (P = 0.004), a RUNX1mut (P = 0.09) by trend, to be EVI1 positive (P = 0.04) & to have higher BAALC (P < .001) expression. A high GPR56 expression associated with worse European LeukemiaNet genetic risk (P = 0.002), a high CD34+/CD38- burden (P = 0.002) & higher cumulative incidence of relapse (CIR, P = 0.04, Figure 1a). Within the low CD34+/CD38- burden pts, high GPR56 expression identified a group of pts with higher CIR compared to pts with low GPR56 expression (P = 0.05, Figure 1b).

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High GPR56 expression associated with higher CIR & worse outcome predictors. In pts with a low CD34+/CD38- burden, high GPR56 expression defined a subgroup with higher CIR after HSCT possibly due to an independent GPR56-defined LIC population. GvL effects after HSCT might be insufficient to control high LIC burden AML & LIC targeting therapies may improve outcome. Disclosure: No conflict of interest disclosed. V1014

Early Replication of Human Cytomegalovirus (HCMV) is associated with a reduced risk of hematologic relapse in myeloid neoplasias after matched Unrelated Stem Cell Transplantation (HSCT) in an anti-thymocyte-globuline dependent manner Beelen D.W.1, Bujor L.-V.1, Stempelmann K.2, Fiedler M.3, Crivello P.2, Trilling M.3, Dittmer U.3, Ditschkowski M.1, Fleischhauer K.2 Universitätsklinikum Essen, Westdeutsches Tumorzentrum, Klinik für Knochenmarktransplantation, Essen, Germany, 2Universitätsklinikum Essen, Institut für Zelltherapeutische Forschung, Essen, Germany, 3Universitätsklinikum Essen, Institut für Virologie, Essen, Germany 1

Introduction: We previously reported that early (i. e. within 100 days) replication of HCMV (HCMVrep) reduces the risk of hematologic relapse (HR) after T-cell-replete allogeneic HSCT in patients (pts) with acute myeloid leukemia (AML) (Elmaagacli A et al. Blood 118:1402-1412,2011). This effect has been confirmed by some, but not all subsequent clinical studies. Among other reasons, this may be particularly due to the administration of ex- or in-vivo T-cell depletion. Hence, we tested the influence of ATG as part of the acute graft-versus-host disease (GvHD) prophylaxis on the efficacy of HCMVrep to reduce the risk of HR after HSCT. Methods: A cohort of 349 pts (median age 55 years [yrs], range 18 -73 yrs) underwent HSCT from 10/10 HLA-matched unrelated donors for AML (n = 292) or myelodysplastic syndromes (MDS, n = 57). HCMVrep was monitored by pp65-antigenemia assay in 236 pts (68%), and in 113 pts (32%) by quantitative polymerase chain reaction, the latter being predominantly applied in pts receiving ATG (77% vs. 23%, p < 0.0001). Pts without or with prophylactic ATG did not differ significantly regarding patient and donor HCMV serostatus (–/– 29% vs. 24%, +/+ 35% vs. 44%, –/+ 8% vs. 6%, +/– 28% vs. 25%). The cumulative incidence (CI) of HCMVrep and of HR were calculated with non-relapse-mortality (NRM) as a competing risk (CR). In multivariate analysis of HR, NRM was also considered as a CR, and HCMVrep was used as a time-dependent covariate. Results: The CI of HCMVrep was 30% (95%-confidence interval [95%CI]: 22%-39%) in pts without compared to 46% (95%-CI: 37%-54%) in pts with ATG-prophylaxis (p < 0.002). The CI of HR was significantly lower for pts without ATG in whom HCMVrep was documented (18%, 95%-CI: 10%-29%) compared to pts without HCMVrep (32%, 95%-CI: 24%-41%)(p < 0.04). In contrast, the CI of HR was comparable in pts with ATG prophylaxis irrespective of HCMVrep (33%, 95%-CI:22%-45%, with and 28%, 95%-CI:19%-37%, without HCMV replication; p = n.s.). Multivariate analysis identified HCMVrep (hazard ratio:0.43, 95%-CI: 0.21-

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0.88, p < 0.02) and chronic GvHD (hazard ratio:0.31, 95%-CI: 0.16-0.64, p < 0.002) as those covariates which independently reduced the risk of HR. Conclusions: This analysis on a larger patient cohort thus confirms our previous report. Abrogation of the HR risk reduction by prophylactic in-vivo T-cell depletion using ATG supports the notion that T cells are important players in the underlying mechanism. Disclosure: No conflict of interest disclosed. V1015

Outcome of allogeneic stem cell transplantation for patients with high-risk acute leukemia according to donor and graftversus-host disease prophylaxis Lindner S.1, Berg T.1, Riemann J.1, Ajib S.1, Jedlickova Z.1, Gueller S.1, Lang F.1, Sackmann A.1,2, Goekbuget N.1,2, Martin H.1, Serve H.1,2, Bacigalupo A.3, Bug G.1 Universitätsklinikum Frankfurt, Frankfurt am Main, Germany, 2German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Heidelberg, Germany, 3Universita‘ Cattolica del Sacro Cuore, Fondazione Policlinico Universitario Gemelli, Roma, Italy 1

Introduction: In high-risk acute leukemia (HR-AL) an allogeneic hematopoietic stem cell transplantation (HSCT) is the only potentially curative treatment and HSCT from alternative donors are increasingly performed. Methods and results: We retrospectively analyzed the outcome of 151 HR-AL patients (AML/ALL, n = 120/31, median age 51 years, range, 1972 and HCT-CI 3, range, 0-11) treated in our transplant center between 1/2011 and 6/2015 according to donor type and graft-versus-host disease (GvHD) prophylaxis: matched related donor using standard immunosuppression (IS) (MRD group, n = 29), matched unrelated donor (MUD group, n = 84) and mismatched (9/10 HLA allele identical) unrelated donor using IS and ATG (MMUD group, n = 25) and haploidentical family donor/mismatched (8/10 HLA-allele identical) unrelated donor using IS and posttransplant cyclophosphamide (PT-Cy group, n = 14). A standard (n = 39), intermediate (n = 61) or reduced-intensitiy (n = 51) conditioning regimen was applied in CR (n = 108) or active disease (n = 43). In summary, patients and disease characteristics were well balanced between groups and all patients engrafted to ≥0.5/nL neutrophils with a median time of 20 days (range, 12-32) and ≥50/nL platelets of 22 days (range, 11-1687+). Cumulative incidences (CI) of acute GvHD G3-4 at day +100 and chronic GvHD at 1 year differed significantly between groups with the highest CI observed in the MRD group (Table 1). With a median follow-up of 29 months (range, 3-60), probabilities of overall survival (OS) at 3 years were 54 ± 10%, 66 ± 6%, 44 ± 11% and 93 ± 7% (p = 0.06). Conclusions: Our results suggest that PT-Cy-based haploidentical transplants are feasible in HR-AL patients and comparison to ATG-based MMUD transplants in a prospective randomized trial is warranted. MRD transplants without ATG were associated with high rates of severe acute and chronic GVHD as previously published.

Fig. 1. OS probability by donor type. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Immuntherapie experimentell V1016

Enhanced engineering of CAR T-cells using non-viral Sleeping Beauty transposition from minicircle vectors Monjezi R.1, Miskey C.2, Gogishvili T.1, Schleef M.3, Schmeer M.3, Hermann E.1, Ivics Z.2, Hudecek M.1 Universitätsklinikum Würzburg, Medizinische Klinik und Poliklinik II, Würzburg, Germany, 2Paul-Ehrlich-Institute, Medical Biotechnology, Langen, Germany, 3 PlasmidFactory, Bielefeld, Germany 1

Tab. 1. CI of TRM, GvHD and relapse by donor type

Introduction: Immunotherapy with T cells modified with gamma-retroviral or lentiviral (LV) vectors to express a chimeric antigen receptor (CAR) has shown remarkable efficacy in clinical trials. However, the potential for insertional mutagenesis and genotoxicity of viral vectors is a safety concern, and their cost and regulatory demands a roadblock for broad clinical translation. Here, we engineer CAR T cells through non-viral Sleeping beauty (SB) transposition of CAR genes from minimalistic DNA vectors called minicircles (MCs) and demonstrate this approach is more effective and has a superior safety profile compared to previous gene-transfer strategies. Methods: We constructed MCs encoding a CD19-CAR and hyperactive transposase SB100X respectively through intramolecular recombination from corresponding DNA plasmids and performed nucleofections into CD8+ and CD4+ T cells of healthy donors. An insertion site library was constructed from polyclonal CAR T cells for massive parallel sequencing on the Illumina MiSeq platform. Results: The stable gene transfer rate and cell viability was 4-fold (n = 7) and 1.5-fold (n = 3) higher when MCs rather than conventional plasmids were used to deliver CAR transposon and SB100X transposase (p < 0.01). In aggregate, this translated into a 6-fold higher yield of CAR+ T cells after 14 days of culture. Importantly, CD19-CAR T cells modified by MC-based SB transposition were equally effective as LV transduced CD19-CAR T cells in vitro and in a murine xenograft model (NSG/Raji-ffLuc), where a single administration of CD8+ and CD4+ CAR T cells lead to complete eradication of lymphoma. To address biosafety, we determined the gene copy number (mean of 5, in CD8+ T cells) and performed the most comprehensive genomic insertion site analysis performed with SB in T cells to date. The data show a closeto-random integration profile of MC-derived CD19-CAR transposons,

Abstracts

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without preference for highly expressed or cancer related genes. Intriguingly, a significantly higher proportion of SB compared to LV integrations had occurred in genomic safe harbors that are not expected to cause genotoxicity (7-fold; p < 0.01). Conclusion: Our data demonstrate for the first time the potential to engineer CAR T cells through SB transposition from MC vectors. The ease-ofhandling MC vectors and superior safety profile compared to viral vectors position our novel approach to become a preferred gene-transfer strategy in advanced cellular and gene-therapy. Disclosure: Razieh Monjezi: No conflict of interest disclosed. Michael Hudecek: Employment or Leadership Position: M.H. and Z.I. are co-inventors on a patent application related to this work; Honoraria: PCT/ US2013/055862 V1017

Functional in vivo monitoring of adoptive T cell therapy via a novel dual-luciferase transgenic mouse model Szyska M.1, Herda S.1, Althoff S.1, D´abundo D.1, Heimann A.1, Russ J.1, Dörken B.2, Arnold R.2, Blankenstein T.3, Na I.-K.1,2 Experimental and Clinical Research Center (ECRC), Berlin, Germany, 2Charité, Department of Hematology, Oncology and Tumor Immunology, Berlin, Germany, 3Max Delbrück Center for Molecular Medicine, Berlin, Germany 1

Introduction: Efficient tumor eradication via adoptive T cell therapy (ATT) requires trafficking of tumor-specific T cells to the tumor, their infiltration into the tumor tissue and activation at the tumor site. Peripheral tolerance, insufficient antigen presentation by tumor cells and local immunosuppression in the tumor microenvironment as well as on-target/ off-tumor toxicity are all limiting the success of ATT. Methods: We generated the transgenic mouse line BLITC (bioluminescence imaging of T cells) expressing an NFAT (nuclear factor of activated T cell)-dependent Click-beetle luciferase and a constitutive Renilla Luciferase, allowing us to monitor migration and activation of transferred T cells in vivo. In order to analyse crucial ATT parameters in a clinically relevant H-Y tumor model, BLITC mice were crossed with HY-TCR transgenic mice and monitored for tumor infiltration and rejection of the H-Y expressing tumor line MB49. Results: We transferred different numbers of CD4 HY-TCR transgenic (Marilyn) BLITC T cells into solid tumor-bearing recipients at least 7 days after tumor injection. Bioluminescence imaging revealed a rapid T cell migration to the tumor draining lymph node and subsequent tumor infiltration within the first week, where T cells were detectable for several weeks. In contrast, the T cell activation signal reproducibly peaked between 5 and 10 days after transfer before rapidly declining. Adoptively transferred T cells inhibited tumor growth in a dose-dependent manner before tumor relapse occurred after 3 to 4 weeks consistent with abrogated T cell activation in all recipients. We further observed lethal GvHD in male recipients receiving as few as 0,5 × 106 Marilyn T cells. Conclusion: In summary, we established a bioluminescent reporter-transgenic mouse model allowing detailed analysis of longitudinal in vivo trafficking and local activation of transferred T cells. This novel tool could be applied in different tumor models in order to develop and evaluate combination and novel therapeutic strategies for optimal tumor-specific ATT overcoming immune suppression by the tumor and limited on-target/ off-tumor toxicity.

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Fig. 1.

Figure 1. Intratumoral flux mice signals upon ATT are shown for activation (CB-NFAT) and migration (RLuc). Tumor growth after ATT is shown. The dashed lines (right graph) indicate remaining mice in each group. The tumor volume was calculated as Ixw2/2. Disclosure: No conflict of interest disclosed. V1018

CD19 CAR-redirected memory stem cell and central memory T lymphocytes generated from naive T cell precursors by inhibition of Akt-signaling induce potent antitumor immunity Hartwig U.F.1, Berger A.1, Weber I.1, Khan S.A.1, Chmielewski M.2, Abken H.2, Theobald M.1 Universitätsmedizin der Johannes Gutenberg-Universität, III. Med. Klinik & Poliklinik – Hämatologie, Internistische Onkologie & Pneumologie, Mainz, Germany, 2Uniklinik Köln, Klinik I für Innere Medizin, Zentrum für Molekulare Medizin, Köln, Germany 1

Background: Adoptive cellular therapy (ACT) of chimeric antigen receptor (CAR)-redirected T cells has advanced as effective therapy for leukemia and solid tumors. However, its efficacy especially to solid tumors is often hampered by limited homeostasis and non durable responses. Recently, stem cell memory T cells (TSCM) that differentiate into central memory (TCM), effector memory (TEM) and effector (TEFF) T cells were shown to elicit potent and long-lived anti tumor responses. Moreover, inhibitors of the PI3K-Akt-mTOR signaling pathway (Inh-VIII) effectively block T cell differentiation and facilitate the generation of TSCM and TCM in vitro. Therefore, in this proof of concept study we investigated the generation of CD19 CAR expressing TSCM/CM from naive precursors by modulating PI3K-Akt-mediated T-cell differentiation to be used for ACT. Methods: Naïve CD45RA+ T cells isolated from PBMC by CliniMACS® were stimulated with CD3/CD28 Dynabeads together with a cytokine

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cocktail and retrovirally transduced with a CD19 CAR 3 days after activation. To promote a TSCM/CM phenotype transduced cells were either αCD3/ CD28 restimulated or cocultured with CD19+ B-ALL (NALM16) together with Akt Inh-VIII. Flow cytometry, IFN-γ ELISPOT and 51Cr-release cytotoxicity assays were performed to assess phenotype and functionality. Redirected T cells were adoptively transferred into NALM-16 engrafted NSG mice to evaluate antitumor reactivity in vivo. Results: In contrast to naive T cells cultured with IL-12 (IL-2), -7, -15, and IL-21, we obtained strong expansion of T cells with a TSCM/CM phenotype upon culture with Akt Inh-VIII. In addition, CD19 CAR+ TSCM/CM elicited strong IFN-γ release and cytotoxicity to NALM-16 cells when compared to mock controls. This effect was also seen in CD19 CAR+ TEM/EFF but less pronounced. Adoptive transfer of 1-5 x106 CD19-CAR redirected TSCM/CM into NALM16 engrafted NSG mice revealed significant leukemia regression in spleen and bone marrow. Antileukemic responses were also seen with CD19 CAR transduced TEM/EFF but TSCM/CM appeared to be more effective when applied at lower numbers (1 x106 cells/mouse) indicating more sustained immunity in this humanized mouse model. Further studies are in progress to elucidate the functional properties of CD19 CAR TSCM/CM. Conclusion: These proof of concept studies demonstrate that ex vivo generated TSCM/CM redirected to leukemia or tumor by retroviral transfer of CARs may be a promising approach to improve ACT. Disclosure: No conflict of interest disclosed. V1019

Induction of NK and T cell immune responses against acute myeloid leukemia (AML) by bispecific NKG2D-CD16 and NKG2D-CD3 fusion proteins Maurer S.1, Körner S.1, Kanz L.2, Grosse-Hovest L.3, Jung G.4, Salih H.R.1 Clinical Collaboration Unit Translational Immunology, German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Partner site Tübingen, Department Hematology and Oncology, Eberhard Karls University, Tuebingen, Germany, 2Department of Hematology and Oncology, Eberhard Karls University, Tuebingen, Germany, 3Synimmune GmbH, Tuebingen, Germany, 4Department for Immunology, Eberhard Karls University, Tuebingen, Germany 1

Monoclonal antibodies (mAbs) targeting tumor cells are meanwhile an established tool for the treatment of many malignancies. The interaction of a classical mAb’s Fc portion with Fcγ receptors (FcγR) on immune effector cells is crucial for its efficacy, but often not capable to induce sufficiently potent antitumor immune responses. Accordingly, novel antibody formats like bispecific antibodies are being developed, which allow for specific activation of defined pools of effector cells including T cells. We recently reported on the development of immunostimulatory bispecific fusion proteins that target ligands of the immunoreceptor NKG2D (NKG2DL) which are widely expressed on malignant cells but generally absent on healthy tissue. Besides the extracellular domain of NKG2D as targeting moiety, our constructs comprise Fab-fragments of either an agonistic CD16 or CD3 antibody to stimulate NK cells or T cells, respectively. Here we provide further data on the preclinical characterization of these compounds. Dose titration assays revealed an increased affinity of NKG2D-CD16 to the FcγR on NK cells as compared to our previously described (e.g. Steinbacher et al, 2014) Fc-optimized NKG2D-IgG1 fusion proteins. This was mirrored by a substantially increased ability to induce lysis of NKG2DL-transfectants and NKG2DL-positive primary AML cells by allogeneic NK cells in a target antigen-dependent manner. The NKG2D-CD3 constructs in turn were found to potently and specifically stimulate allogeneic and autologous CD4+ and CD8+ T cells. Next we comparatively analyzed the efficacy of T cells and NK cells to lyse autologous leukemia cells upon treatment with NKG2D-CD3 and NKG2D-CD16, respectively, by using PBMC of AML patients directly obtained ex vivo at time of diagnosis in long term cytotoxicity assays. Already NKG2D-CD16 potently induced AML cell lysis, and this was, in line with their higher effector potential, by far exceeded upon stimulation of T cells with NKG2D-CD3. Notably, the CD3-NKG2D construct also potently induced

Abstracts

proliferation of T cells, which constitutes an important asset to treat patients with higher tumor burden. Taken together, we here report on novel “antibody-like” bispecific constructs that take advantage of the highly tumor-restricted expression of NKG2DL and potently activate the reactivity of NK or T cells for immunotherapy of leukemia. Disclosure: No conflict of interest disclosed. V1020

HLA ligandome analysis of different hematological malignancies identifies a small panel of naturally presented “pan-leukemia” antigens Stickel J.S.1, Kowalewski D.J.2, Walz S.2, Schuster H.2, Berlin C.2, Schemionek M.3, Brümmendorf T.3, Vucinic V.4, Niederwieser D.4, Kanz L.1, Salih H.R.1,5, Weisel K.1, Rammensee H.-G.2,6, Stevanovic S.2,6 Universitätsklinik Tübingen, Hämatologie und Onkologie, Tübingen, Germany, Universität Tübingen, Abteilung für Immunologie, Tübingen, Germany, 3 Universitätsklinikum RWTH Aachen, Abteilung für Hämatologie, Onkologie, Hämostaseologie und SCT, Aachen, Germany, 4Universitätsklinikum Leipzig, Abteilung für Hämatologie und Onkologie, Leipzig, Germany, 5Clinical Cooperation Unit Translational Immunology, German Cancer Consortium (DKTK), DKFZ partner site Tübingen, Tübingen, Germany, 6German Cancer Consortium (DKTK), DKFZ partner site Tübingen, Tübingen, Germany 1 2

Effective T-cell based cancer immunotherapy requires exact knowledge of tumor-associated antigens (TAAs) that can act as rejection antigens. While the current paradigm views mutation-derived neo-antigens as the most promising targets, we have recently demonstrated that T-cell responses target panels of non-mutated TAAs in patients with hematological malignancies. Using the approach of direct HLA ligandome analysis we were able to identify and characterize several immunogenic TAAs for CLL (Kowalewski, PNAS 2015), AML (Berlin, Leukemia 2014), multiple myeloma (MM) (Walz, Blood 2015) and CML. In this project, we analyzed our HLA ligandome data for the existence of “pan-leukemia” antigens to allow for a broad application in immunotherapy approaches for the treatment of hematological malignancies. To identify leukemia-exclusive HLA ligands we compared the HLA ligandomes of CLL (HLA class I (I) n = 35, HLA class II (II) n = 30), AML (I n = 19, II n = 20), MM (I n = 15, II n = 12) and CML (I n = 16, II n = 15) with our normal tissue database including 153 (I) and 82 (II) ligandomes of various healthy tissues (e.g., blood, bone marrow, spleen, kidney, liver, brain). Overlap analysis revealed only 0.6% (16/2716, I) and 0.3% (10/3141, II) of the identified leukemia exclusive antigens to be represented across all analyzed hematological malignancies. These “pan-leukemia” antigens (n = 26) include some highly interesting targets associated with T-cell activation (HSH2D), lymphoid development (IL2RF) and oncogenesis (LYN protooncogene, RAB5A). However, none of the “pan-leukemia” antigens shows frequent representation (>20%) in all entities. More similarities could be identified within the subgroups of lymphoid and myeloid malignancies: A total of 60 (8.6%, I) and 35 (6.9%, II) leukemia-exclusive antigens were shared among CML and AML, while CLL and MM shared 249 (14.6%, I) and 52 (7.4%, II) of exclusive antigens, respectively. For reference, randomized intra-cohort dichotomization of MM ligandomes exemplarily detected mean intra-entity overlaps of 13.2% (I) and 8.4% (II), respectively. Taken together our approach of direct HLA ligandome analysis of hematological malignancies identified a small panel of “pan-leukemia” antigens that show representation in all analyzed entities. However, due to the uneven frequencies of these antigens across hematological malignancies, target identification for immunotherapy should still be performed in an entity-specific or even patient-individual manner. Disclosure: No conflict of interest disclosed.

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V1021

Immune checkpoint inhibition increases specific immune responses for leukemia-associated antigens against myeloid leukemic cells Greiner J.1,2, Goetz M.2, Schneider V.2, Schrezenmeier H.3, Wiesneth M.3, Bullinger L.2, Doehner H.2, Hofmann S.2 Diakonie-Klinikum Stuttgart, Hematology and Oncology, Stuttgart, Germany, University of Ulm, Department of Internal Medicine III, Ulm, Germany, 3 University of Ulm and German Red Cross Blood Donor Service BadenWürttemberg – Hessen, Institute of Transfusion Medicine, Ulm, Germany 1 2

Immunotherapy in cancer treatment experienced a breakthrough in the last few years. The efficacy of immunotherapeutic approaches e.g. immune-checkpoint inhibitors, chimeric antigen receptor T cells (CARs) or bi-specific T cell activating antibodies becomes more and more evident. However, mechanisms of these immune responses and responsible antigen structures have to be further elucidated. Leukemia-associated antigens (LAAs) represent immunogenic antigens that are target structures relevant for elimination of malignant cells by cytotoxic T cells (CTL) and are candidates for specific immunotherapy. In this work, we investigated the influence of PD-1 antibody Nivolumab and CTLA-4 antibody Ipilimumab on the antigen-specific immune responses by specific T cells against leukemic myeloid blasts in functional T cell assays using ELISpot Assays, tetramer-analysis and colony-forming immunoassays. We investigated T cell responses stimulated against known LAAs like RHAMM, PRAME, WT-1, SSXIP2, Proteinase 3, Survivin, Aurorakinase A and NPM1 against several AML cell lines and samples of AML patients. Expression of different LAAs were measured and correlated to functional T cell assays. In colony forming unit immunoassays, T cells stimulated against various LAAs indicated a significant inhibition of CFUs in AML patient samples. In all patient samples, effectors activated against at least one LAA were successful to decrease the colony number. The LAAs PRAME, RHAMM and WT1 showed highest frequency and intensity of immunogenic reactions: PRAME stimulated CTL induced an immune response in 83% of tested samples in CFUs, stimulated with WT1 in 75% and with RHAMM in 58% of colonies from AML samples. Specific immune responses of cytotoxic T cells were also detected by ELISpot assays and correlated to results detected in CFUs. Immune effects increased adding nivolumab to CFU whereas no effect was measured when incubated with Ipilimumab. The combination of Nivolumab and Ipilimumab showed no additional effect of immune responses compared to Nivolumab alone. Taken together, the immune checkpoint inhibitor Nivolumab increases specific T cell responses of LAA-stimulated cytotoxic T cells and the cytotoxic effect of T cells against blasts of AML patients. No additional effect was detected with Ipilimumab. These data suggest that PD-1 antibodies could be an immunotherapeutic approach in AML and combination with LAA-directed vaccination strategies are interesting options. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Gastrointestinale Tumoren, Pankreaskarzinom V1022

Clinical validation of response and resistance factor candidates to targeted therapy in gastric cancer (GC) Haffner I.1, Luber B.2, Maier D.3, Geier B.3, Kretzschmar A.4, Fischer von Weikersthal L.5, Ahlborn M.6, Riera Knorrenschild J.7, Rau B.8, Weissinger F.9, Fuxius S.10, Neumann S.11, Decker T.12, Schierle K.13, Wittekind C.13, Lordick F.1 University Hospital Leipzig, University Cancer Center Leipzig (UCCL), Leipzig, Germany, 2Technische Universität München, Institute of Pathology, München, Germany, 3Biomax Informatics AG, Planegg, Germany, 4MVZ Mitte, Leipzig, Germany, 5MVZ, Amberg, Germany, 6Braunschweig Community Hospital, Braunschweig, Germany, 7University Hospital Marburg, Department of Hematology and Oncology, Marburg, Germany, 8Charité Universitätsmedizin Berlin, Department of General, Visceral, Vascular and Thoracic Surgery, Berlin, Germany, 9Evangelisches Krankenhaus Bielefeld gGmbH, Klinik für Innere Medizin, Hämatologie/Onkologie und Palliativmedizin, Bielefeld, Germany, 10Onkologische Schwerpunktpraxis Heidelberg, Heidelberg, Germany, 11Klinikum Wolfsburg, Medizinische Klinik II, Wolfsburg, Germany, 12 Studienzentrum Onkologie Ravensburg, Ravensburg, Germany, 13University Hospital Leipzig, Institute of Pathology, Leipzig, Germany 1

Background: 10–20% of GC overexpress HER2, a membrane-bound receptor tyrosine kinase (RTK) which belongs to the epidermal growth factor receptor (EGFR) family. Drugs directed against HER2 have shown mixed success in the treatment of advanced GC. While trastuzumab, a monoclonal antibody addressing HER2 has been approved for 1st-line treatment of stage IV HER2+ GC, trastuzumab-emtansine failed to improve outcomes in 2nd-line and lapatinib, a small molecular RTK inhibitor of HER2 and EGFR was not effective in 1st- and 2nd-line. Until now, primary and secondary resistance against HER2-directed treatment of GC is not well understood. The VARIANZ study, which is part of the SYS-Stomach consortium (supported by the German Federal Ministery of Education and Research, BMBF) aims to assess resistance mechanisms in gastric cancer samples from patients receiving trastuzumab. Methods: In this multicenter study, patients who receive medical treatment for advanced GC are recruited in 31 German sites. The HER2 status is verified centrally by two dedicated GI pathologists (KS and CW) using immunohistochemistry (IHC, DCS, HI608C0I) and chromogenic-in-situ hybridization (CISH, Zytomed Systems, C-3022-40). In a second step, samples will be used to validate resistance factors that are identified as interesting candidates by in-vitro and in-silico modelling within the consortium. Results: From 26 May 2014 to 31 March 2016, we have enrolled 261 patients in this ongoing project (74% male, median age 64 years). At present, 217 samples were fully characterized for the HER2 status. According to criteria from the Trastuzumab for Gastric Cancer (ToGA) study, 47 of 217 samples were characterized HER2+ by central testing. In 30 samples that were diagnosed as HER2+ by local pathologists the HER2 status could not be verified centrally. 7 HER2- probes in local testing were characterized as HER2+ by central testing. The overall deviation rate between local and central testing is 31%. Conclusions: HER2-expression in GC is heterogeneous and still not easy to assess. Variability between local and central HER2 assessment is significant. Robust biomarkers predicting resistance to HER2 and other target therapies are needed. Updated results will be shown at the meeting. Disclosure: No conflict of interest disclosed.

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V1023

Diagnostic and prognoses of relapses of gastric and esophageal adenocarcinoma within follow-up care – a retrospective analysis Tauchert F.K.1, Strauß-Rothenbücher A.2, Bankstahl U.S.3, Heidsieck T.4, Reichart A.1,3, Tameizi W.3, Jäger E.1, Al-Batran S.-E.3 Krankenhaus Nordwest / UCT Frankfurt, Klinik für Onkologie, Hämatologie & Palliativmedizin, Frankfurt am Main, Germany, 2Krankenhaus Nordwest / UCT Frankfurt, Zentralinstitut für Radiologie und Neuroradiologie, Frankfurt am Main, Germany, 3Krankenhaus Nordwest / UCT Frankfurt, Institut für klinisch onkologische Forschung, Frankfurt am Main, Germany, 4Krankenhaus Nordwest / UCT Frankfurt, Klinik für Allgemein-, Visceral- und minimal invasive Chirurgie, Frankfurt am Main, Germany 1

Introduction: Depending on the tumor stage, surgery with or without perioperative chemotherapy is the standard of care for gastric and esophageal adenocarcinoma (AEG). However there is a high risk of relapse. Due to the non-existence of studies to estimate the value of standard follow-up care (FUC) the guidelines in Europe are quite different. Methods: All patients who started the FUC within a study (FLOT-3, FLOT-4, FLOT65+ & HER-FLOT) at our center were included in this analysis. The patients studied were those with either histologically or radiologically determined relapse or those with highly probable clinical course of relapse. Two groups were compared. The first group comprised the patients whose recurrence was detected by routine follow-up prior to the development of clinical signs (asymptomatic group (group I); the second group consisted of the patients who developed clinical symptoms due to a recurrence that was detected afterwards (symptomatic group (group II)). The main parameters were the time until recurrence occurred, treatment and survival. The ct-scans have been reviewed to show which method would have been sufficient to show the relapse. Results: Out of 69 patients entering the FUC 31 (44,9%) had a relapse. 12 (38,7%) were in group I and 19 (61,3%) in group II. The time until relapse occurred was comparable (12 vs. 8,5 months). In group I 4 patients received a potential curative treatment and 8 palliative chemotherapy, none best supportive care alone. In group II one patient received potential curative treatment, 14 palliative chemotherapy and 4 best supportive care. Median survival after relapse was longer in group I (41 vs. 13,5 months). Retrospectively, the abdominal ct scan alone would have been able to detect 27 relapses (87,1%); but 2 relapses were detected in thoracal ct only, one was able to receive potential curative treatment afterwards. One patient wasn’t detected with ct scan but with endoscopy and received potential curative treatment afterwards. Conclusion: Intensive FUC with regular ct scans is able to detect a substantial number of asymptomatic relapse of AEG. Asymptomatic relapse is associated with higher overall survival and some patients are able to receive potential curative treatment afterwards. Abdominal ct scans alone is able to detect most relapses but can miss local relapses, which might be treated curatively as well Disclosure: No conflict of interest disclosed. V1024

Wnt/β-catenin-signalling molecules in the Barrett’s esophagus – an in-depth analysis Thieme R.1, Götzel K.1, Chamnitzer O.1, Lyros O.1, Lamprecht A.-K.1, Rolfs F.1, Gockel I.1 Universitätsklinikum Leipzig, Klinik und Poliklinik für Viszeral-, Transplantations-, Thorax- und Gefäßchirurgie, Leipzig, Germany 1

To improve esophageal adenocarcinoma (EAC) therapy and to investigate cellular pathways responsible for EAC development in-depth will be fascinating challenges for the next years. To improve our understanding of EAC biology and treatment is needed because EAC is increasing over the last decades in the western population and the overall survival remains poor. The Wnt/β-catenin-signalling pathway has been studied in various malignancies, but its role in carcinogenesis of EAC remains illusive.

Abstracts

Wnt/β-signalling molecules were characterized by RT- and qRT-PCR using 6 immortalized cell lines representing the different stages of the Barrett’s sequence from squamous epithelium to EAC (epithelial (EPC1, EPC2), metaplastic (CP-A), dysplastic (CP-B) and adenocarcinoma (OE19, OE33)). The ligands Wnt3a and Wnt5a were only expressed in the epithelial cells EPC1 and EPC2. With progression of the Barrett’s sequence Wnt3a and Wnt5a were dramatically decreased or absolutely absent beginning from the metaplastic stage. However, the dysplastic cell line CP-B shows a very high Wnt5a level, but its functional or pathophysiological contribution is still not clear. Investigating the 10 frizzled receptors, we could detect FZD1-3 and FZD5-7 in all cell lines, FZD4 and FZD8-10 show a cell type specific pattern, missing FZD8 in EAC cells. However, FZD10 is only found in EAC cells. The Wnt-signalling target gene Axin2 is increasing within the Barrett’s sequence. Wnt3a (200ng/mL) treatment in EAC cells results in an increase of Axin2 expression in OE19 cells but not in OE33 cells. However, an increase in GSK3ß-phosphorylation was only seen in the dysplastic cell line CP-B and not in squamous epithelial and EAC cells. An in-depth analysis of Wnt/β-signalling molecules including ligands, receptors, co-receptors and downstream show the high diversity of this signalling pathway in the Barrett’s sequence from squamous epithelium to EAC. Our findings suggest a strong contribution for the Wnt-axis with Wnt-depending and -independent stages during the tumorigenesis of the esophageal adenocarcinoma. Supported by the Junior Research Grant of the Faculty of Medicine, University of Leipzig Disclosure: No conflict of interest disclosed. V1025

Age and chemotherapy dose intensity in patients with gastrointestinal cancer Mahler S.1, Trautmann K.1, Rentsch A.2, Ehninger G.1, Folprecht G.1 Universitätsklinikum Carl Gustav Carus, Medizinische Klinik I, Dresden, Germany, 2Universitätsklinikum Carl Gustav Carus, Universitätskrebszentrum, Dresden, Germany 1

Background: For gastrointestinal cancer, patients (pts) in clinical trials have a median age that is approximately 10 years (y) lower than the age of pts treated in clinical trials. There are few data describing the tolerability of therapeutic regimens outside of clinical studies. Methods: Based on the institutional data base of the University Hospital Dresden (Germany) that contained all chemotherapy doses since January 2002, the time from first dose to first dose reduction and the treatment duration with ≥ 90% were analyzed depending on the age, treatment schedule and disease. Results: In total, 857 oxaliplatin or irinotecan based doublet regimens (with or without addition antibodies) were given for colorectal cancer and 305 treatment courses with cisplatin/weekly infusional 5-FU or neoadjuvant epirubicine based therapy for treatment of gastric cancer. Compared to pts with an age < 60 y, the time to first dose reduction was similar for elderly pts receiving oxaliplatin based regimen (pts 60–70 years: HR 1.03 [95% CI: 0.70–1.53], pts >70 y. HR 1.34 [0.74–2.41], pts >75 y: HR 2.46, [95% CI: 0.99–6.16]). Pts >70 y. receiving an irinotecan based regimen had earlier dose reductions than pts <60 y (HR 2.01 [95% CI: 1.22–3.29], p < 0.01). This effect was not observed in pts with an age of 60-70 y (HR 0.97 [0.65–1.43]). For gastric cancer pts receiving cisplatin based therapy, first dose reduction was significantly earlier in pts between 60 and 70 y (HR 3.79 [1.96–7.33]., p < 0.01) and for pts >70 y (HR 3.63 [1.53–8.62], p < 0.01). In elderly pts treated with neoadjuvant epirubicine base therapy, chemotherapy was terminated after a median of 5 weeks resulting in incomplete therapy. Conclusion: The age dependent tolerability of different chemotherapy doublets depends on the schedule and seems to be decreased with cisplatin or irinotecan. Our retrospective study suggests that an age (and comorbidity-) based dosing schedules should be developed.

Oncol Res Treat 2016;39(suppl 3):1–327

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Disclosure: Sandra Mahler: No conflict of interest disclosed. Gunnar Folprecht: Financing of Scientific Research: Roche/Genentech, Merck, Lilly, Sanofi-Aventis, Baxalta, Bayer, Boehringer; Expert Testimony: Merck V1026

Patient-reported outcomes of patients with advanced or metastatic pancreatic cancer in German outpatient centres – first data from the sub-project PanLife of the clinical Pancreatic Cancer Registry (TPK) Hegewisch-Becker S.1, Wolf T.2, Aldaoud A.3, Scheiner-Sparna R.4, Hamm D.4, Marschner N.5, for the TPK Registry Group Onkologische Schwerpunktpraxis Hamburg Eppendorf, Hamburg, Germany, BAG/Onkologische Gemeinschaftspraxis, Dresden, Germany, 3Dr. Aldaoud – Dr. Schwarzer Forschungsgesellschaft mbH, Leipzig, Germany, 4iOMEDICO, Freiburg i. Br., Germany, 5Praxis für interdisziplinäre Onkologie & Hämatologie, Freiburg i. Br., Germany 1 2

Introduction: Pancreatic cancer is the fourth leading cause of cancer death in Europe. About 80% of patients (pts) are diagnosed with locally advanced or metastatic disease (mPC). 5-years survival rate for these pts is less than 2%. The main treatment goals in these patients are the improvement of tumor symptoms and the stabilization of quality of life (QoL) on a level as high as possible since survival can be prolonged only marginally. PanLife prospectively investigates patient-reported outcomes (PRO) to better understand the real-life situation of pts with mPC in German routine practice. Methods: The Tumour Registry Pancreatic Cancer (TPK) is a prospective, multicentre, observational study of pts with mPC receiving systemic palliative therapy. Starting in February 2014, 101 study sites in Germany recruit pts at the start of their 1st-line treatment. In the sub-project PanLife PROs are measured with the validated questionnaires EORTC QLQ-C15PAL and EORTC-QLQ-PAN26. Pts are asked to fill in these questionnaires at the start of 1st-line treatment (T0) and every two months thereafter. Here, interim data on quality of life (QoL) and symptoms of pts at T2 (four months after start of 1st-line treatment) will be reported for all pts recruited into the registry until data cut-off June 2016 (800 pts expected). Results: Preliminary results from data cut-off June 2015 (451 pts): At the start of palliative treatment median age was 70 years (range 43-94 years), 56% of pts were male. The most frequent 1st-line treatments were gemcitabine and nab-paclitaxel (42%), gemcitabine monotherapy (27%), and FOLFIRINOX (18%). Questionnaire return rate was 84% at start of 1stline treatment. At the time of this analysis, 229 pts had returned the T2 questionnaire (four months after start of therapy). About one third of pts reported deterioration, improvement or no changes in QoL (global score EORTC QLQ-C15-PAL), respectively. About half of the pts reported improvements in fatigue, dysgeusia, weakness, changes in bowel habits and being bothered by side effects. About a third of pts reported improvements in nausea and vomiting, pain, and pancreatic pain. Conclusion: Four months after start of 1st-line treatment improvements in QoL and PROs are reported by about half of pts with mPC. Updated data will be presented for pts subgroups, e.g. pts treated with different treatment regimens. Disclosure: No conflict of interest disclosed. V1027

Targeting Nrf2 and Hemeoxygenase-1 (HO-1) to treat esophageal adenocarcinoma cells in vitro Thieme R.1, Rolfs F.1, Klotz A.1, Gockel I.1 Universitätsklinikum Leipzig, Klinik und Poliklinik für Viszeral-, Transplantations-, Thorax- und Gefäßchirurgie, Leipzig, Germany 1

The esophageal adenocarcinoma (EAC) is characterized by an increasing incidence in the western population by about 600%. Despite improved therapeutically options the overall survival of patients with advanced EAC is still poor. Cellular detoxification mechanisms are essential for cell’s survival and could be a vulnerable target to combat EAC. Thereby Nrf2, which is responsible for the cellular stress defense by inducing hemeoxygenase-1 and

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glyoxalse-1 by an antioxidant response element, will be evaluated as a potential target in EAC therapy. Using an in vitro model representing the Barrett’s sequence from squamous epithelium to EAC, the epithelial (EPC1, EPC2), metaplastic (CPA), dysplastic (CP-B) and adenocarcinoma (OE19, OE33) cell lines were used, to investigate Nrf2, HO-1, GLO1 and GLO2 expression by RT- and qRT-PCR and the responsiveness of EAC cells to 5-fluorouracil (5-FU) in a Nrf2 and HO-1 dependent manner. The transcription factor Nrf2 and its downstream targets GLO1, GLO2 and HO-1 are increased with the Barrett’s sequence from squamous epithelium to EAC. OE33 cells are highly receptive to 5-FU with an IC50 of 0.58µM, no growth inhibitory effect was seen in OE19 cells under 5-FU treatment. Inhibiting Nrf2 with trigonelline (0.1µM or 1µM) drop down the IC50 to 5.7µM of 5-FU in OE19 cells. In OE33 cells no further reduction of the IC50 could be observed by trigonelline treatment. However no additional inhibitory effect by affecting the HO-1 by ZnPP under 5-FU treatment could be observed. 5-FU slightly increased GLO1 in OE33 and OE19 cells (1.5fold) after 48h. HO-1 was increased in 1µM and 10µM 5-FU treated OE33 after 24h, 48h and 72h by 5.8fold. 5-FU had no effect on HO-1 expression in OE19 cells. The Nrf2 itself was slightly increased in 5-FU treated OE33 cell after 24h and 48h. We could show different expression levels in the cellular detoxifying enzymes GLO1, GLO2 and HO-1 in the EAC cells as well as differences in the responsiveness to 5-FU in those cells. A targeted inhibition of a central transcription factor Nrf2 resulted in an increased responsiveness to 5-FU in a before 5-FU-resistent cell line. Further target identification and inclusion of other chemotherapeutics will help to find new therapeutic target in EAC treatment. Disclosure: No conflict of interest disclosed.

Freier Vortrag

Urogenitale Tumoren V1028

Subgroup analyses and updated overall survival from the phase 2 trial of Lenvatinib (LEN), Everolimus (EVE), and LEN+EVE in metastatic Renal Cell Carcinoma (mRCC) Hutson T.E.1, Dutcus C.E.2, Ren M.2, Baig M.2, Fishman M.3 Texas Oncology, Dallas, United States, 2Eisai Inc., Woodcliff Lake, United States, H.Lee Moffitt Cancer Center and Research Institute, Tampa, United States

1 3

Introduction: In the randomized phase 2 study of LEN, EVE, and LEN+EVE in mRCC, LEN+EVE significantly improved the primary endpoint median progression-free survival (PFS) vs EVE (14.6 vs 5.5 months [mos]; hazard ratio [HR] 0.40; 95% confidence interval [CI] 0.24–0.68; P = 0.0005). LEN alone (7.4 mos) also improved median PFS vs EVE (HR 0.61; 95% CI 0.38–0.98; P = 0.048). We report PFS outcomes by patient subgroups. Methods: Patients with progressive clear cell mRCC following 1 VEGF-targeted therapy received LEN (24 mg/d), EVE (10 mg/d), or LEN+EVE (18+5 mg/d). Exploratory subgroups included baseline tumor size (sum of tumor diameters), Memorial Sloan-Kettering Cancer Center (MSKCC) risk group, and metastasis site. The data cutoff for the primary analysis was June 13, 2014; updated cutoff (for overall survival [OS] only) was July 31, 2015. Results: Median PFS was improved in LEN+EVE vs EVE in all examined subgroups (Table). In an updated analysis of OS, the median was 25.5 mos (95% CI 16.4–32.1), 19.1 mos (95% CI 13.6–26.2), and 15.4 mos (95% CI 11.8–20.6) for LEN+EVE, LEN, and EVE, respectively. In comparison with EVE, OS was longer for LEN+EVE, a trend that was maintained through the cutoff for the updated OS analysis (HR 0.59; 95% CI 0.360.96; P = 0.065). Notably, the HR remains < 0.6, and the upper limit of the CI no longer crosses 1.

Abstracts

CONTENTS AUTHOR INDEX

Conclusions: The PFS advantage with LEN+EVE vs EVE was maintained in all examined subgroups. The updated OS analysis demonstrated a trend towards survival benefit with LEN+EVE vs EVE in patients with mRCC following 1 VEGF-targeted therapy. Tab. 1 to V1028. Median PFS, mos

MSKCC risk Favorable Intermediate Poor Baseline tumor size < Median ≥ Median Metastasis site Bone Visceral organs Lymph nodes

Abstracts

Median PFS, mos LEN n n = 52

Disclosure: Thomas Hutson: Advisory Role: Pfizer, Novartis, BMS, Eisai, Bayer, Jansen, Astellas; Financing of Scientific Research: Pfizer, Novartis, BMS, Eisai, Bayer, Jansen, Astellas; Expert Testimony: Pfizer, Novartis, Eisai, BMS; Other Financial Relationships: Speakers bureau: Pfizer, Novartis, BMS, Eisai, Bayer, Jansen, Astellas Mayer Fishman: Advisory Role: Eisai, Alkermes; Honoraria: Patent pending; Expert Testimony: BMS, Exelixis, Eisai, Pfizer, Merck, Prometheus, Bayer; Other Financial Relationships: Speakers Bureau: Pfizer, GlaxoSmithKline; NCCN, M2GEN

LEN+EVE n = 51

EVE n = 50

HR (95% CI); P-value (LEN+EVE vs EVE)

11 18 23

18.4 7.2 5.6

12 19 20

20.1 14.6 5.6

12 19 19

9.8 5.5 3.5

0.25 (0.08–0.76); p = 0.009 0.35 (0.15–0.80); p = 0.0243 0.44 (0.20–0.98); p = 0.0936

27 25

7.4 7.2

24 27

14.7 11.2

25 25

5.5 5.3

0.34 (0.16–0.71) p = 0.0035 0.39 (0.19–0.80) p = 0.0134

23 49 32

7.2 7.4 8.0

16 41 29

5.4 9.5 14.7

17 44 31

3.6 5.5 5.5

0.41 (0.18–0.94) 0.44 (0.26–0.77) 0.28 (0.14–0.58)

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V1029

Active surveillance (AS) as potential treatment modality for management of patients (pts) with metastatic renal cell carcinoma (mRCC)? Ivanyi P.1, Eggers H.1, Seidel C.2, Ganser A.1, Grünwald V.1 Medizinische Hochschule Hannover, Klinik für Hämatologie, Hämostaseologie, Onkologie u. Stammzelltransplantation, Hannover, Germany, 2 Universitätsklinikum Hamburg Eppendorf, Klinik für Onkologie, Hämatologie, Knochenmarktransplantation mit Sektion Pneumologie, Hamburg, Germany 1

Background: AS might reflect a feasible treatment approach in mRCC patients (pts). Data on pts selection criteria for AS remain elusive. Therefore, we explored pts who were assigned to AS in our clinic to identified predictive characteristics. Methods: mRCC pts treated at our center with first line (1stL) treatment (Rx) initiation and/or primary diagnosis of mRCC from 04/2000 - 06/2012 were retrospectively analyzed. Data were extracted from pts records. Pts. receiving 1stL Rx deferred by ≥ 6 months after mRCC diagnosis were considered as AS pts. Descriptive statistics, Kaplan-Meier and cox-regression were applied. Results: 222 mRCC pts at a median age of 62 (range (r): 26-86) years, predominantly male (68.9%), whereof 94.3% received a nephrectomy revealing in 80.2% a clear cell RCC were identified. 68 AS pts. were identified (median time to1stL Rx:21.4 (r:6-199) months). Overall-survival (OS) from 1stL Rx was superior in AS pts (AS: 32.7 (23.9-41.5) vs. non-AS: 24.6 (17.3-31.9) months, log-rank: p = 0.003). Time from diagnosis of mRCC to 1stL Rx, an initial N-positive stage, and time from RCC to mRCC ≤ 1 year were identified as independent risk parameter for OS. Conclusion: AS seems to be feasible in selected patients. Delay of therapy had no negative impact on OS and was applied in 30.6% of pts at our center. Ultimately, AS defers toxicity and could lead to improved quality of life. mRCC with aggressive behavior achieve an inferior outcome and may not represent the target population for AS. Additional studies on AS are warranted. Disclosure: No conflict of interest disclosed. V1030

Recognizing symptom burden in advanced prostate cancer: a global patient and caregiver survey Schostak M.1, Oh W.K.2, Tombal B.3, Delacruz A.4, Tomlinson B.5, Ripley A.V.6, Drudge-Coates L.7, Mastris K.8, O’Sullivan J.M.9, Shore N.D.10 Universitätsklinikum Magdeburg, Klinik für Urologie und Kinderurologie, Magdeburg, Germany, 2Tisch Cancer Institute-Icahn School of Medicine at Mount Sinai, New York, United States, 3Université Catholique de Louvain, Institut de Recherche Clinique, Louvain-la-Neuve, Belgium, 4Memorial SloanKettering Cancer Center, New York, United States, 5CancerCare, New York, United States, 6Nielsen, New York, United States, 7King’s College Hospital NHS Foundation Trust, London, United Kingdom, 8Europa UoMo, The European Prostate Cancer Coalition, Essex, United Kingdom, 9Queen’s University School of Medicine, Belfast, United Kingdom, 10Carolina Urologic Research Center, Myrtle Beach, United States 1

Background: Symptoms may indicate disease progression in patients (pts) with advanced prostate cancer (aPC) but may not be recognized or communicated to health care providers (HCPs). Methods: To assess the impact of symptoms on daily life and how aPC pts communicate symptoms to their HCPs, the largest aPC pt survey to date (International Prostate Cancer Symptom Survey) was conducted online and by telephone by Harris Poll in 11 countries (Brazil, France, Germany, Japan, Italy, Netherlands, Singapore, Spain, Taiwan, UK, USA) from Feb 12 to Oct 27, 2015. Because sample sizes varied among countries, cell weighting was used to bring each country’s pts and caregivers to be equal in their respective totals. Results: Overall, 927 aPC pts and 400 caregivers were surveyed. Globally, 99% of pts with bone metastases experienced ≥1 symptom. Of pts with bone metastases, most had symptoms of fatigue (87%), pain in specific areas (83%), and generalized body pain (72%). Moreover, 73% of pts with

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bone metastases noticed pain before receiving a diagnosis of aPC (40% had pain for ≥7 months and 33% for <7 months). Most aPC pts (56%) were uncertain if the pain was cancer-related, 55% felt they had to live with daily pain, 45% sometimes ignored pain, 39% had difficulty talking about pain and 29% did not agree that managing pain was essential to effectively treat cancer. A majority of pts said they would be motivated to report pain if it helped increase quality of life (QoL, 65%) or longevity (57%), kept cancer from worsening (54%), or decreased pain (53%). Caregivers report accompanying pts to 70% of HCP visits and 50% of pts rely on caregivers to ask the most important questions. Pts with a caregiver were more likely than those without to say pain is discussed at every visit (45% vs 32%, P = 0.031). Significantly more caregivers agreed with the statement that pain somewhat or strongly limited patients’ activities (72% of caregivers vs 54% of patients; P < 0.0001). Conclusions: Symptoms in aPC are often under recognized. Tools encouraging effective communication between pts/caregivers and HCPs on early symptom reporting and treatment may lead to earlier symptom identification and control. Enhanced discussion of approved therapies may improve QoL and patient outcomes. Disclosure: No conflict of interest disclosed. V1031

Analysis of overall survival by number of radium-223 injections received in an international expanded access program (iEAP) Wirth M.1, Saad F.2, Keizman D.3, O’Sullivan J.M.4, Carles J.5, Gillessen S.6, Thellenberg Karlsson C.7, Miller K.8, Tucci M.9, Paganelli G.10, Procopio G.11, Gratt J.12, Seger M.12, Nilsson S.13, Heinrich D.14 Universitätsklinikum Carl Gustav Carus Dresden, Klinik für Urologie, Dresden, Germany, 2University of Montreal Hospital Centers, Montreal, Canada, 3 Meir Medical Center, Kfar Saba, Israel, 4Centre for Cancer Research and Cell Biology, Queen’s University Belfast, Belfast, United Kingdom, 5Vall d’Hebron University Hospital, Barcelona, Spain, 6Kantonsspital St. Gallen, St. Gallen, Switzerland, 7Umeå University, Umeå, Sweden, 8Charité Berlin, Berlin, Germany, 9 San Luigi Hospital, Orbassano (Turin), Italy, 10IRST-IRRCS Cancer Center, Meldola, Italy, 11Fondazione IRCCS Istituto Nazionale dei Tumori, Milano, Italy, 12 Pharmaceuticals Division of Bayer, Whippany, United States, 13Karolinska University Hospital, Stockholm, Sweden, 14Akershus University Hospital, Lørenskog, Norway 1

Background: In ALSYMPCA, 6 injections (inj) of radium-223 (Ra-223) at 50 kBq/kg v placebo, improved overall survival (OS) in patients (pts) with metastatic castration-resistant prostate cancer (mCRPC). Early identification of pts likely to receive the planned 6 inj of radium-223 is important. Here are OS analyses by number of Ra-223 inj from iEAP and ALSYMPCA, including prognostic factors associated with 1-4 v 5-6 inj, respectively. Materials and methods: The open-label, single-arm, phase 3b iEAP provided Ra-223 to bone-predominant mCRPC pts. OS was analyzed post hoc by number of Ra-223 inj (1-4 v 5-6) and compared with ALSYMPCA data. Stepwise logistic and stepwise Cox regression analyses of number of inj and OS, respectively, were done. Results: Baseline characteristics for iEAP v ALSYMPCA pts are quite similar. In iEAP, median (med) OS was 6.3 mo in pts with 1-4 inj and not reached with 5-6 inj. In ALSYMPCA, med OS was 6.2 mo in pts with 1-4 inj and 17.9 mo with 5-6 inj. In a logistic regression analysis from iEAP, receipt of 5-6 inj was associated with less pain (none-mild v moderate-severe; P < .0001), lower ECOG score (0-1 v ≥2; P = 0.0081), lower PSA level (med < 141 µg/L v ≥141 µg/L; P < .0001), and higher Hgb (≥10 g/dL v < 10 g/dL; P = 0.0227) at baseline. Stepwise Cox regression analysis showed that 5-6 inj was associated with longer OS (P < .0001). Conclusions: These exploratory analyses suggest that pts with less pain, lower ECOG score, lower PSA level, and higher Hgb level at baseline are more likely to receive 5-6 Ra-223 inj. A prospective randomized trial is needed to confirm a causal relationship between number of inj and OS. Disclosure: No conflict of interest disclosed.

Abstracts

CONTENTS AUTHOR INDEX

V1032

Muscle invasive urothelial carcinoma (MIUC) subclassification with a Nanostring gene panel for survival prediction and exploration of subtype-specific drug targets Rinaldetti S.1, Rempel E.2, Worst T.2,3, Steidler A.3, Weis C.-A.4, Hartmann A.5, Bolenz C.6, Erben P.3 Medizinische Fakultät Mannheim der Universität Heidelberg, III. Medizinische Klinik, Hämatologie und Onkologie, Mannheim, Germany, 2German Cancer Research Center (DKFZ), Heidelberg, Germany, 3Medizinische Fakultät Mannheim der Universität Heidelberg, Klinik für Urologie, Mannheim, Germany, 4 Medizinische Fakultät Mannheim der Universität Heidelberg, Pathologisches Institut Mannheim, Mannheim, Germany, 5Universitätsklinikum Erlangen, Pathologisches Institut, Erlangen, Germany, 6Universitätsklinikum Ulm, Klinik für Urologie, Mannheim, Germany 1

Background: Transcriptome expression studies identified distinct bladder cancer subtypes of prognostic relevance and closely related with breast cancer molecular subclasses. Here we developed a Nanostring-based subtype-screening in a chemotherapy naive patient cohort with MIUC and analysed the subtype-specific expression of drug targets with the prospect of personalizing therapy options. Methods: FFPE samples of MIUC were collected from a cohort of 47 patients (median age: 66 years, m/w: 34/13) treated exclusively with radical cystectomy and bilateral lymphadenectomy. 80 genes were selected using two approaches. First, genes were selected from the prognostic gene signatures published previously (Choi et al., Cancer Cell, 2014: basal, luminal and p53-like). In a second step, we reanalyzed previously published microarray data with gene set enrichment analysis. Subsequently gene expression was measured with the Nanostring nCounter technology. The subclassification was performed by unsupervised hierarchical clustering using the Pearson and Ward methods. The subgroup specificity of potential drug targets (not included in the clustering) was estimated by the Mann-Whitney-U test. Results: Unsupervised hierarchical clustering separated the tumors into 3 clusters: basal (B), luminal (L) and infiltrated (I). These subtypes present a distinct OS (L: 24 mo, B: 87 mo, I: 93 mo, p = 0.002) and DSS (L: 38 mo, B: 108 mo, I: 128 mo, p = 0.017) over a follow-up period up to 12 years. Our study also confirms a highly subtype-specific expression of drug targets. Genes like ERBB2 (HER2neu), ERBB3 and the FGFR gene family are exclusively expressed in the luminal subtype (p < 0.03). The androgen receptor seems only suppressed in the basal subtype (p < 0.001). The infiltrated group shows an isolated overexpression of PDGFRB (p = 0.02). Conclusions: Beside the risk stratification and survival prediction, this nCounter-based screening allows a concise exploration of drug targets. Hereby we demonstrated that the MIUC subtypes show different phenotypes in terms of Tyrosine kinase and hormone receptor expression. Subclassification of MIUC is crucial in future translational studies in order to pave the way to personalized medicine. Disclosure: No conflict of interest disclosed. V1033

Improved outcome of patients with metastatic germ cell tumors: results from a cohort study at two centers in Munich Hentrich M.1, Debole J.2, Gerl A.2 Rotkreuzklinikum, III. Medizinsiche Abteilung, München, Germany, Onkologische Praxis, München, Germany

1 2

Introduction: Treatment of metastatic germ cell cancer (GCC) is based on the International Germ Cell Cancer Collaborative Group (IGCCCG) prognostic classification published in 1997. 5-year survival rates were reported to be 91%, 79%, and 48% for patients with good, intermediate and poor prognosis, respectively. However, treatment results may have improved over time due to cumulative experience. Further, it remains unclear whether there is still a need for dose intensification for intermediate and poor risk patients.

Methods: The records of all patients (pts) with metastatic GCC treated at two institutions in Munich between 2000 and 2013 were reviewed with regard to time of initial diagnosis, histopathology, stage, tumor marker levels, metastatic spread, type and duration of chemotherapy (CT), and outcome. Progression-free survival (PFS) and overall survival (OS) were estimated with the Kaplan-Meyer method. The Log-rank test was used to compare survival distributions of different groups. Results: Of 255 patients identified, 30 pts were exluded due to incomplete data. The median age was 35 years, seminoma (S) and nonseminoma (NS) were diagnosed in 72 (32%) and 153 (68%) pts, and 204 pts (91%) had a primary gonadal GCC. 175 (78%), 30 (13%) and 20 pts (9%) had good, intermediate and poor prognosis according to the IGCCCG classification system. 189 of 225 pts (84%) included into the study were treated as outpatients and 36 (16%) as inpatients. The vast majority of pts received 3 to 4 cycles of platinum-based CT while primary high-dose CT was applied to 3 pts in the poor prognosis group. The 2-year-PFS of pts with good, intermediate and poor prognosis was 91%, 83% and 37%, and the 5-year-OS was 98%, 96%, and 66%, respectively. There was no significant difference in the 5-year-OS between pts in the good and intermediate prognosis group. Conclusions: Compared to data from the 1997 IGCCCG classification system, the outcome of pts with metastastic GCC has considerably improved. Notably, no significant differences in the 5-year-OS were observed between pts with good and intermediate prognosis. While the outcome of pts with intermediate-prognosis is excellent, treatment results in the poor-prognosis group remain unsatisfactory. Disclosure: No conflict of interest disclosed.

Plenarsitzung

Zurück in die Zukunft V1034

The 5,300-year-old Helicobacter pylori genome of the Iceman Maixner F.1, Krause-Kyora B.2, Turaev D.3, Herbig A.4, Krause J.4, Nebel A.2, Moodley Y.5, Rattei T.3, Zink A.1 EURAC research, Institute for Mummies and the Iceman, Bozen, Italy, 2Kiel University, Kiel, Germany, 3University of Vienna, Wien, Austria, 4MPI Jena, Jena, Germany, 5University of Venda, Thohoyandou, South Africa 1

The stomach bacterium Helicobacter pylori is one of the most prevalent human pathogens. It has dispersed globally with its human host, resulting in a distinct phylogeographic pattern that can be used to reconstruct both recent and ancient human migrations. The modern H. pylori strain found in most Europeans (hpEurope) has putatively originated from recombination of the two ancestral populations Ancestral Europe 1 and 2 (AE1 and AE2). However, there exist different hypotheses about when and where the hybridization took place, reflecting the complex demographic history of Europeans. In this study, we screened biopsy samples from the gastrointestinal tract of the Iceman, a 5300-year-old European Copper Age mummy, for the presence of H. pylori. By using metagenomic diagnostics and targeted genome capture, we determined the presence of H. pylori and reconstructed its complete genome. Comparison with contemporary H. pylori sequences and proteomics analysis classified the ancient H. pylori as cytotoxic type strain that triggered already calprotectin release as a result of host inflammatory immune responses. Comparative analysis of ancient housekeeping gene fragments with a global multilocus sequence typing (MLST) database and comparative whole-genome analyses assigned the 5,300-year-old bacterium to the modern population hpAsia2 commonly found in Central and South Asia. The “Iceman” H. pylori is a nearly pure representative of the bacterial population of Asian origin (AE1) that existed in Europe before hybridization, suggesting that the African population (AE2) arrived in Europe within the past few thousand years, which is much later than previously proposed. Disclosure: No conflict of interest disclosed.

Abstracts

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Pflegetagung

Patientenschulung und -beratung

Assessing the information needs of patients with hematologyoncology malignancies: a cross sectional study using the EORTC- QLQ-INFO25-questionnaire Zimmermann N.1 Universitätsklinik Freiburg, Klinik für Innere Medizin 1, Freiburg, Germany

Introduction: Understanding what cancer patients need to know is vital to ensure quality care. Nurse-patient communication in oncology presents a big challenge in oncology care. Little is known about German cancer patients’ need for information. The purpose of this cross-sectional study was to assess the information needs of patients at in- and outpatient oncology wards. Method: The EORTC Quality of Life (QOL) Group has developed an instrument to assess the quality and quantity of information received by patients in the areas disease, medical tests, treatment, other services, different places of care and how to help themselves, as well as aspects as helpfulness of and satisfaction with this information. The study included adult patients diagnosed with a hematology-oncology malignancy. A convenience sample of 150 patients, from 1st-15th of April were asked to fill in the QLQ-INFO25 questionnaire. 132 of them responded (88%). Demographic and clinical data were collected and a descriptive statistic was performed using SPSS 23.0. Result: 83% of all respondents felt the received information moderate to very helpful. 76% of the patients are satisfied with the amount of received information. At least 52 patients want more information. Women wish to receive more information compared to men. All in all more than half (58%) of the participants don´t want to receive more information. The top three priorities relate to effects on sexual activity, different places of care and additional help outside the hospital. Older adults (>60 years) do not prioritize different information needs compared with younger patients. Both are very satisfied with information about medical treatments as well as their diagnosis. Patients newly diagnosed (< 6 months) with hematologic cancer are not as satisfied (32% answered little to not at all) with the received information as long-term patients. Counting the averages of replies, it shows outpatient cancer patients received more information compared to inpatient patients. Conclusion: This study reports the results of needs from a selected group of cancer patients. Not the amount of information is essential for improving the quality of cancer patients care but the personalized information exchange between health professionals and patients. Limitation is that cancer patients do not know what could be possible. Keywords: hematological patients, oncology care, information needs, EORTC QLQ- INFO25 questionnaire Disclosure: No conflict of interest disclosed.

Nursing consultation and instruction program for the prophylaxis of oral mucositis treated with chemotherapeutics König P.1 Hochschule Furtwangen, Fakultät Gesundheit, Sicherheit, Gesellschaft, Furtwangen, Germany 1

A damage of the oral mucosa is a wellknown sideeffect of stomatotoxic chemotherapie. Symptoms can appear some days after application of the chemotherapie. The dimention of mucosa damage and discomfort could be pretty variably. Many studies focused the effects of various substances to treat oral mucositis. Furthermore there are several studies focusing interventions for prevention oral mucositis, but they showing only some benefit with mostly low degree of evidence. This study should contribute to clarify the role of guidance in preventing oral mucosits. The results of this study should help to enhance the preparation of patients before treating with chemotherapie.

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The main result of the clinical study was that no significant effects of the intervention program in relation to the occurrence of oral mucositis could be demonstrated (p = 0.766). Participants in the control group as well as in the test group reported a significant deterioration of life quality regarding the mouth. The intensity of mouth hygiene measures between the baseline measurement and the last measurement increased moderately in the control group and significantly in the test group. Thus, the „Mund-Protect“ program clearly leads to a greater increase in mouth hygiene measures compared to the control group. The interview data show that the patients found the supervision, the information provided at an early stage, and the advice in crisis situations very helpful. Disclosure: No conflict of interest disclosed.

Neue Therapien – ärztliche und pflegerische Aspekte I New therapies in multiple myeloma Goldschmidt H.1 Universitätsklinikum Heidelberg, Medizinische Klinik, Innere Medizin V, Heidelberg, Germany 1

For a long time (1960 - 2000) the only treatment options for multiple myeloma were chemotherapy, glucocorticoids and high-dose therapy. The first new drug introduced in the therapy of multiple myeloma was Thalidomid. Immunmodulatory drugs (IMIDS) have been derived from Thalidomid. Lenalidomid and Pomalidomid, both IMIDS, are more active than Thalidomid and have other side effects. Another mode of action in MM is proteasome inhibition. Successful evaluated proteasome inhibitors are Bortezomib, Carfilzomib and Ixazomib. Combinations of the new drugs are very effective. Nowadays we have the opportunity to use monoclonal antibodies in multiple myeloma. Another option is to use histone deacetylase inhibitors (HDAC) in relapsed multiple myeloma. Treatment options and side effects will be presented and discussed. Disclosure: No conflict of interest disclosed.

Selbstmanagement «Work & Care» – Reconciling work and caregiving in the context of cancer Jähnke A.1, Bischofberger I.1 Careum Forschung, Kalaidos Fachhochschule Gesundheit, Zürich, Switzerland

Family carers, who look after ill or disabled significant others, play an essential role in health care provision nationally and internationally. Since 2007 Careum Research has been conducting the R&D program «work & care». Applied research activities in various populations and care settings aim to identify challenges and best practice of reconciling employment and family caregiving. In the context of cancer, it is widely acknowledged that family carers are crucial in supporting their family members, particularly in fluctuating home care phases. Surveys conducted for Swiss employers show, that health care professionals take on private care responsibilities more often than employees in other economic sectors. This dual role is called «Double-Duty Caregiving». Little is known about the crossover between professional and private commitment, and its interferences and consequences for work and care. Data from two different research projects will be presented focusing on reconciling work and caregiving in the context of cancer. Firstly, data from 13 in-depth interviews with working carers of cancer patients conducted from 2011 - 2012 in Switzerland. MaxQDA® was used for content analysis. Secondly, data from 30 narrative interviews conducted from 07/2015 - 03/2016 using a Grounded Theory approach with theoretical sampling will be discussed based on ongoing data analysis. During acute or latent phases, flexible work and care arrangements, as well as accountable workmates and health professionals were crucial for

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reconciliation. In end-of-life care, family carers highlighted relieving and stabilizing effects of a sensitive workplace, and the social support provided by workmates. This supportive workplace culture enhanced to fulfill demanding care responsibilities, and to cope with grief and ‚return back to life’ after the family member’s death. Given the fluctuating nature of cancer, health and social service professionals should refer to illness trajectory models when tailoring their advice and support to working family carers. Also, sensitizing at the workplace, notably human resource staff and management is pivotal for preventing crises in family carers’ private and work life. Because of their specific insights and experiences, double-duty caregivers can represent highly valuable resources for health care teams and institutions by making the voices of the ill family members heard. For further information: http://www.careum.ch/workandcare/double-duty-caregiving Disclosure: No conflict of interest disclosed.

“Self care for caregivers” Mori H.1 MFA – Medial Viktor Frankl Association Vienna, Psychotherapeutische Praxis, Wien, Austria 1

Caregivers have to find a balance between a very challenging profession and their own carefulness about their personal and private values in life. Being interested in a supportive and meaningful work-life-balance is a vital necessity for staying healthy and happy while despite fulfilling a very intensive dedication for a profession that is strongly identified with human aspects of life. From birth until the end of a patients life caregivers are very demanded to give a lot of attention to the suffering people while at the same time they should make a job in a highly professional medical quality. To train a healthy form of selfmanagement it is helpful and essential to use concepts of the humanistic psychotherapies. Meaning in life, identity and authenticity as well as autonomy are important factors that induce energetic support. (Viktor E. Frankl). The concepts of “Fitness” (Gerhard Uhlenbruck) and “Salutogenesis” (Aaron Antonovsky) are valuable approaches to a good life and help the “helpers” to be good professionals as well as to have a sufficient armament to protect the personal “oecological system” (Harry Merl) in finding meaning in their work as well as acknowledgements and in the consequence creating a self esteem that enables the caregivers to be satisfied in their jobs. Oncological medical and health care is very stressful and needs special attention for the caregivers to care about themselves too. Disclosure: No conflict of interest disclosed.

Palliativpflege

Aufgabenbereich die Koordination verschiedenster Versorgungsleistungen. Das SAPV-Team gewährleistet eine 24-h Rufbereitschaft an sieben Tagen in der Woche sowie ein Multiprofessionelles Ärzte-und Pflegeteam, welches in regelmäßigen Teamsitzungen einen umfassenden Informationsstand der aktiv betreuten Patienten erwirbt. Disclosure: No conflict of interest disclosed.

Early integration of palliative care Köhler A.1 Jena University Hospital, Palliativmedizin, Jena, Germany

Aim: We want to provide an overview about the work and the chances of a palliative care service integrated as an early additional care in the in-patient care of the department of Oncology / Hematology / Bone morrow transplantation at the University Hospital of Jena. Background: In patients with a non-curative treatable cancer diagnosis, different trials demonstrate a benefit of patients and their relative, when a specialized palliative care service is integrated early and in a structured way into the oncological care (Bakitas 2015, Zimmermann 2014, Temel 2010). Aims of an early integrated palliative care service for patients treated as in-patients in an oncological ward are: (a) improvement of quality of life through regular and structured assessment and treatment of symptoms and needs, (b) maintenance of resources of the patients, (c) sensitive communication to improve acceptance of diagnosis, (d) psychosocial support, e.g. biography works, (e) support of relatives, (f) discharge management and (g) advanced care planning. In addition the palliative care service provides support and training for the staff of the oncological wards, to improve the shared care of the patients. Patients care is provided through a multi-professional team. Team members are palliative care physicians, palliative nurses, physiotherapists, occupational therapist, psychologist, social workers and ministers and volunteers. Shared aim of all team members is to provide care based on the wishes and needs of the patient. The specialized palliative care service for in-patients treated in the deparement of oncology, hematology and bone-morrow-transplantation was established 2.5 years ago. The talk present experiences and results from this period. Disclosure: No conflict of interest disclosed.

Spezialisierte onkologische pflegerische Versorgung Nurse Navigators Cancer Care - “The red thread” in the cancer care system Kern Fürer C.1

Specialized outpatient palliative care

Menniger A.

Introduction: Cancer patients are treated increasingly interdisciplinary, through long treatment processes in various disciplines and are cared for by a variety of professional groups. The numerous interfaces make the supply chain vulnerable to interruptions. In addition, the communication and coordination between all stakeholders becomes increasingly challenging. These challenges hinder the implementation of a patient centered care system. In order to overcome this issue. International literature identifies the assignment of a key contact person in the oncological treatment process, the so called Nurse Navigator. A Nurse Navigator improves the cooperation and communication between the people involved and the coordination of care. The main goal of the master thesis was to define the tasks and competences of Nurse Navigators, which are necessary for the optimization of communication and coordination within the patient-centered care in multimodal assisted cancer patients.

Universitätsklinikum Jena, Klinik für Innere Medizin II, Palliativmedizin, Jena, Germany 1

Mein Name ist Ariane Menniger und ich arbeite als Pflegefachkraft Palliativ Care im SAPV-Team in Jena. Mein Aufgabenbereich beinhaltet in erster Linie der Versorgung unheilbar erkrankter Menschen mit einer Palliativ-medizinisch relevanter Symptomatik sowie deren Angehörige in ihrer gewohnten Umgebung. Ziel meiner Arbeit ist es, die Selbstbestimmung sowie die Lebensqualität möglichst zu erhalten und zu verbessern. Meine Arbeit erfolgt auf der Grundlage einer Versorgungsstruktur, welche sich auf die Palliativversorgung spezialisiert. Das bedeutet, dass unsere Patienten sowohl eine palliativärztliche als auch eine palliativpflegerische Versorgung erhalten. Ebenso gehört zu meinem

Abstracts

Spital Walenstadt, Walenstadt, Switzerland

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Methods: By means of a systematic literature search in the databases PubMed, Cochrane, CINAHL and PsycINFO, eleven studies were analyzed and summarized. To refine the results of the research literature and to gain additional insights as well as to clarify contradictions, five experts have been interviewed using a semi-structured interview guideline. The results of both surveys were compared and analyzed. Results: Nurse Navigators need to take care of two core tasks: “accompanying transitions during vulnerable phases” and “maintaining continuity”. They facilitate coordination and communication, provide psychosocial support and education. They take over the representation of the interests of patients and demonstrate presence. These are significant components of a patient-centered care, which contribute to an optimized cooperation in a multidisciplinary team. To achieve these goals, a high level of technical, methodological, social and personal skills is needed. Conclusion: The analysis reveals specific areas of responsibility, which positively influence the communication and coordination within the multidisciplinary teams. Additionally, the required competence profile is derived. Disclosure: No conflict of interest disclosed.

First year of a central consultation service for oncology nursing within the Center of Integrated Oncology (CIO) at the University Hospital Cologne Rütjes A.1, Schneider E.1 Uniklinik Köln, Centrum für Integrierte Onkologie (CIO), Köln, Germany

Introduction: In October 2015 we began to establish a central consultation service for oncology nursing on all wards of the Centre for Integrated Oncology (CIO) at the University Hospital of Cologne. The rationale for establishing this central structure are requirements of the Deutsche Krebsgesellschaft, asking for a minimum of nurses specialized in oncology nursing in each department, but there was a constant lack of qualified nurses. Methods: Two specialized oncology nurses (one with a bachelor degree) work part time (50%) for the oncology consultation service. They are in charge for all departments of the Center of Integrated Oncology (CIO). In order to keep up with relevant issues of nursing practice, they work the other 50% on a ward. The first two years of the project are financed by the Deutsche Krebshilfe.The main target group is nurses on all wards, particular on wards without specialized nurses. Teams are able to call the consultation service in case of a severe nursing related problem. Thus the consultation service supplies nurses with evidence based information and discusses nursing intervention concerning a particular patient in need. The service can be called by “Orbis”, telephone or e-mail. The consultation service visits all wards weekly on the basis of a structured scheme. Nursing care visits, short teaching lessons during shift reports and individual training on the job with new team members are offered. Results: The first year we focused on systematic rollout. 23 wards of 16 departments will be included by September 2016. Information needs relate primarily on new treatments and supporting nursing interventions. The main obstacles are inconsistent nursing procedures in the different departments and lack of time of nursing staff. Factors of success are a culture of mutual trust and confidence, availability on request and provision of helpful information. The central structure of the consultation service is efficient and supports quality development of nursing practice. Conclusion: There is a great need for evidence based information on nursing interventions throughout the wards. The consultation service proofs a successful structure to transfer knowledge to the wards and constantly improve nursing practice. Disclosure: No conflict of interest disclosed.

Specialized nursing for patients with oral cancer drugs

Praxis Dr. Vehling-Kaiser, Med. Fachangestellte für Onkologie, Landshut, Germany 1

Ten years ago there was a far-reaching change in the antiproflierative treatment. Meanwhile there are a lot of new concoctions in the Imatinib and monthly new are added. By now 20% of the tumour treatment is made up of the oral antiprofilerative substances. At first sight it seems like that those oral concoctions offer many advantages, e.g. less visits of the medical practices, no infusions and the easy taking of pills. But if we take a closer look at the oral treatment, we will be confronted with some problems we haven´t expected at the beginning. Some years ago the Adagio-survey has already represented, that only 14% of the patients take the oral treatment adequately. The problems connected with this are obvious: less affect of the intended treatment and a larger spectrum of adverse effects.In an oncological medical practice the control of the adherence and coaching of the patients take a lot of time. That´s why we have to build up new structures within the oncological medical practice. To this for example the delegation of medical activities to specially trained staff belongs to. One example for such an approach is the arrangement of a so called consultation-hour for oral and subcutaneous treatment of tumour (in short: TKI-consultation-hour). Some assumptions for the arrangement of such a consultation-hour are: 1. Indemnification of trained employees. Those for example are medical assistants or caregivers who have finished the basic class Oncology but also the class to become a medical assistant for oral and subcutaneous treatment of tumour and therefore have a lot of experience with the daily handling of TKI`s. 2. The assistants should get an own room for their consultation-hours, where undisturbed conversations are possible. 3. The control of the adherence and the coaching of the patients aren´t possible while being engaged with other job-related activities just as activities in the chemo-room for example. 4. It is necessary to develop standardised control mechanisms with the help of appropriate forms and to complete evaluations. 5. If it´s possible medical practices additionally should command a mobile oncological service in order to give patients, who can´t attend the medical practice, the safety by close-meshed controls. Disclosure: No conflict of interest disclosed.

Klangschalenmassage / Klangschalenmeditation Singing Bowls meditation and singing bowls massage Kleinsimon I.1 Universitätsklinikum Jena, Klinik für Innere Medizin II, Palliativmedizin, Jena, Germany 1

Mein Name ist Ingrid Kleinsimon und ich arbeite als Krankenschwester auf der Palliativstation des Universitätsklinikum Jena. Unseren Patienten biete ich Mediation mit Einbeziehung von Klangschalen und die Klangschalenmassage an. Was bedeutet das: Bei einer Klangschalenmassage werden speziell gefertigte Klangschalen verschiedener Grundfrequenz und unterschiedlicher Größen auf den bekleideten Körper aufgesetzt und angeschlagen oder direkt über den Körper gehalten, ohne ihn zu berühren. Auf diese Weise überträgt sich der Schall des erzeugten Tons auf den Körper. Dies wird als Vibration im Körper wahrgenommen. Diese Anwendung gibt der Seele Kraft, kann sehr entspannend und beruhigend wirken. Blockaden können gelöst und Schmerzzustände gelindert werden. Diese Methode spricht einen großen Personenkreis an. Angehörige können in die Klangschalenbehandlung mit einbezogen werden. Es ist eine ganzheitliche Methode. Wir machen eine gemeinsame Meditation und ich werden praktische Beispiele benennen. Patienten nehmen das Angebot dieser Anwendung sehr gern an. Disclosure: No conflict of interest disclosed.

Damnali G.1

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Ekel und Scham Shame and disgust Müller D.1 Klinikum Nürnberg, Palliativstation, Nürnberg, Germany

Every day nurses engage in the privacy and intimacy of patients. They care for individuals who are dependent on help in the activities of everyday life, cause personal hygiene by, treat malodorous tumors, dispose of vomit and other secretions such as stool or urine. They often exceed their own limits or those of another and emotions like disgust or shame can be caused. These feelings have an impact on nurses, patients and their relationship. Therefore, they need to deal with the knowledge and skills with shame and disgust to protect themselves and the patient and would treat with each other. Disclosure: No conflict of interest disclosed.

Ergotherapie in der Palliativmedizin Occupational therapy in palliative care Loster A.1 Universitätsklinikum Jena, Institut für Physiotherapie, Jena, Germany

Die Ergotherapie bedient sich komplexer motivierender und handlungsorientierter Methoden und Verfahren, die gemeinsam mit dem Patienten ausgewählt werden. Dabei werden handwerklich/kreative, spielerische und funktionelle Techniken wie auch lebenspraktische Übungen eingesetzt. Die Vorgehensweise ist stets ganzheitlich: angesetzt wird bei Stärken und Fähigkeiten des Patienten, nicht an seinen Schwächen. Ziel ist es, Eigeninitiative und Motivation des Patienten zu fördern, ihn psychisch zu stär-

Abstracts

ken und ihm im Alltag zu größtmöglicher Selbständigkeit zu verhelfen. Scheinbar nebenbei wird an seinen kognitiven, motorischen und sensorischen Defiziten gearbeitet. Der Fantasie von Patient und Therapeut sind in Bezug auf die Inhalte, die Medien und Materialien der Therapie kaum Grenzen gesetzt. Im Workshop soll die ergotherapeutische Arbeit auf der Palliativstation Jena, wie auch in der Palliativmedizinischen Komplexbehandlung auf Normalstationen des Universitätsklinikums Jena anhand von Fallbeispielen vorgestellt werden. Disclosure: No conflict of interest disclosed.

Fort- und Weiterbildung, Akademisierung, Zukunftsperspektiven “Example for research project” - Benefit of empirical research in a further education Programme Mattern K.1, Dreischer T.2 Universitätsklinikum, Dresden, Germany, 2Universitätsklinikum Carl Gustav Carus der TU, Dresden, Germany 1

Recent developments in professional care in Germany, including the pending common foundation training and the increasing academic influence on nursing education, raises the question of whether there are any benefits to students conducting research projects during specialised further education programmes. The presentation discusses a project involving participants from a further education programme, using this example from practice to highlight the benefits that such a project may offer. Disclosure: Katrin Mattern: Employment or Leadership Position: im Universitätsklinikum Dresden Tanja Dreischer: No conflict of interest disclosed.

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Studententag

Berufliche Perspektiven in Hämatologie & Onkologie

(The results are part of a thesis submitted by M.S. to Charité – Universitätsmedizin Berlin) Disclosure: No conflict of interest disclosed.

Research and science Fischer T.1 Otto-von-Guericke Universität Magdeburg, Hämatologie und Onkologie, Magdeburg, Germany 1

In this workshop, we will discuss research and science opportunities for young doctors in Hematology and Oncology within Germany. A selection of the spectrum of structurized programs as MD/PhD programs, “Else-Kröner-Forschungskollegs” for clinician scientists, “Gerok”-positions, MD-programs for clinician scientist within collaborative research centers (“Sonderforschungsbereiche”, “Graduiertenkollegs”), etc. in Germany will be presented. In addition, this workshop will also present extra-mural funding opportunities in Germany to foster scientifc careers of young clinician scientists. Disclosure: No conflict of interest disclosed.

Promotionsstipendien

Sieglinde Welker-DGHO-Promotionsstipendium Characterization of hSNM1B/Apollo´s role within the Fanconi anemia/BRCA DNA repair pathway Schmiester M.1, Salewsky B.1, Demuth I.1 Charité – Universitätsmedizin Berlin, Berlin, Germany

Introduction: Fanconi anemia (FA) is a genetic disorder clinically characterized by congenital defects, increased risk of cancer and bone marrow failure. To date, there are 19 known genes that encode FA proteins (FANCA-FANCT), all of which function in the FA/BRCA DNA repair pathway. FA cells exhibit hypersensitivities towards DNA interstrand crosslink (ICL)-inducing agents. Recent research suggests that reactive aldehydes may be the primordial sources of these DNA lesions in vivo. We recently showed that the nuclease hSNM1B/Apollo interacts with FANCP/SLX4. Here, the interacting domains were mapped. Furthermore, the response of cells depleted of hSNM1B/Apollo or/and FANCP/SLX4 towards acetaldehyde exposure was tested. Methods: Co-immunoprecipitation experiments with endogenous, ectopically expressed and in vitro-translated hSNM1B/Apollo and FANCP/ SLX4 were performed. hSNM1B/Apollo- or/and FANCP/SLX4-depleted U2OS-cells (siRNA) were treated with acetaldehyde and used in colony survival assays and chromosome breakage analyses. Results: While the interaction of hSNM1B/Apollo with endogenous and ectopically expressed FANCP/SLX4 was confirmed, it could not be shown with in vitro-translated proteins. Mapping of the interacting domains suggested at least two separate binding areas. After acetaldehyde exposure, the ability to form colonies remained unchanged in hSNM1B/Apollo-depleted cells and decreased mildly in cells depleted of FANCP/SLX4 and of both proteins. The amount of chromosome breaks per cell increased, however, there was no significant difference between the control cells and the knockdown cells. Conclusion: The results demonstrate that hSNM1B/Apollo is directly linked to the FA/BRCA pathway via a complex interaction with FANCP/ SLX4. The normal or only mildly decreased acetaldehyde tolerance of hSNM1B/Apollo- and FANCP/SLX4-depleted cells, respectively, stands in contrast to the known hypersensitivity of both these cells towards ICL-inducing agents and of some other FA cells towards acetaldehyde. While these findings raise questions regarding the primary DNA-damaging mechanism of this particular aldehyde, they also suggest that cells deficient in the FA/ BRCA pathway vary in their chemical sensitivities depending on their specific mutation. Studying these differences will contribute to a better evolutionary and mechanistic understanding of the pathway.

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José Carreras-DGHO-Promotionsstipendium Serum of myeloproliferative neoplasms and upon aging comprises soluble factors that affect proliferation and differentiation of hematopoietic stem and progenitor cells Lubberich R.K.1, Walenda T.1, Goecke T.W.2, Strathmann K.3, Koschmieder S.4, Wagner W.1 Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Stem Cell Biology and Cellular Engineering, Aachen, Germany, 2 Department of Obstetrics and Gynecology, RWTH Aachen University Medical School, Aachen, Germany, 3Institute of Transfusion Medicine, RWTH Aachen University Medical School, Aachen, Germany, 4Department of Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, Faculty of Medicine, RWTH Aachen University, Aachen, Germany 1

Introduction: Myeloproliferative neoplasms (MPN), such as polycythemia vera (PV), essential thrombocythemia (ET) and primary or post-PV/post-ET myelofibrosis (MF), are causally linked to cell intrinsic somatic mutations. In this study we analyzed if serum of MPN patients comprises soluble factors that affect proliferation and differentiation of hematopoietic stem and progenitor cells (HPCs) in vitro. Methods: CD34+ HPCs from umbilical cord blood of healthy donors were cultured in medium supplemented with serum samples (10%) of 8 PV patients, 15 ET patients, 13 MF patients, or 15 healthy blood donors. After five days of culture, cell division (as an indicator of proliferation) and preservation of immature hematopoietic surface markers was assessed using flow cytometric analysis of carboxyfluorescein succinimidyl ester (CFSE) retention and CD34, CD133 and CD45 expression. Colony forming unit (CFU) frequency was determined after 7 days. To further analyze effects of normal aging on hematopoiesis supportive factors, we compared 15 healthy young (< 25 years) and 15 old (>50 years) donors. DNA methylation changes were analyzed in MPN patients by pyrosequencing and Infinium HumanMethylation450 BeadChip array. Results: Serum from MF, PV and ET patients significantly enhanced proliferation (p < 0.001, p < 0.001 and p < 0.001, respectively) and MF and ET sera additionally increased the expression of CD34. The number of CFUs was higher with serum of MF, PV or ET patients as compared to healthy controls (p = 0.01; p < 0.001and p < 0.001). Addition of serum from healthy young versus old blood donors did not evoke differences in proliferation or immunophenotype of HPCs, whereas the CFU potential was significantly increased by serum from elderly patients (p = 0.02). We demonstrated that epigenetic age prediction using our 3CpG Epigenetic-Aging-Signature (Weidner et al., Genome Biol., 2014) is likewise applicable to cell free DNA in serum and we are currently applying this to MPN serum samples. Global methylation analysis of DNA from peripheral blood revealed 73 CpGs that differ significantly in MF versus healthy controls, and only 5 CpGs varied between PV and MF (adjusted p-value < 0.05). Conclusion: Serum of MF and ET patients comprises soluble factors that activate proliferation, support a primitive phenotype, and support the CFU potential of CD34+ cells in vitro. Such feedback mechanisms may activate the residual normal hematopoiesis or enhance development of the disease.

Abstracts

CONTENTS AUTHOR INDEX

Disclosure: Richard Lubberich: No conflict of interest disclosed. Wolfgang Wagner: Stock Ownership: WW is cofounder of Cygenia GmbH, which can provide service for the 3CpG age prediction model.; Honoraria: RWTH Aachen University has applied for a patent for the 3 CpG age prediction model

José Carreras-DGHO-Promotionsstipendium Anti-leukemic microRNA-9 down-regulates the oncogenic transcription factor ERG, sensitizes to standard chemotherapy& associates with improved outcome in acute myeloid leukemia (AML) Grimm J.1, Bill M.1, Weidner H.1, Jentzsch M.1, Knyrim M.1, Schmalbrock L.1, Schubert K.1, Gerloff D.1, Wurm A.1, Cross M.1, Lange T.1, Behre G.1, Niederwieser D.1, Schwind S.1 Abteilung Hämatologie und Internistische Onkologie, Universitätsklinikum Leipzig, Leipzig, Germany 1

In AML high expression of transcription factor ERG is associated with worse outcome. miRs targeting ERG might exhibit anti-leukemic functions. 3 miR-9 binding sites (BS) were identified in the ERG 3’-untranslated region & analyzed by luciferase assay. AML cell lines with low miR-9 & high ERG level (KG1a, KASUMI-1) were transfected with miR-9 expression or empty control vector or treated with Paclitaxel (PTX) -a potential miR-9 inducer. In K562 cells (low ERG & high miR-9 level) knock-down (KD) was achieved by anti-miR-9 transfection. We analyzed mRNA & protein levels by qRT-PCR & western blot & performed proliferation assays. Findings were validated in primary AML blasts of 3patients (pts).The meth-

Abstracts

ylation of miR-9 promoter regions was assessed by bisulfite conversion followed by qRT-PCR. Bone marrow (BM) pri-miR-9 expression in 115 AML pts was examined. For outcome analyses the highest quartile defined high miR-9 expressers. After miR-9 overexpression in 293T cells, 1 of the miR-9 BS showed down-regulation of luciferase activity to 52.0% (P = 0.02) that was rescued by BS mutation. miR-9 overexpression reduced ERG mRNA & protein levels in KG1a (65% & 20%) & KASUMI-1 (69% & 60%). KD of miR-9 in K562 cells resulted in increased ERG expression (mRNA 450%, protein 320%). Higher miR-9 expression had anti-leukemic effects by inhibiting proliferation of KG1a & KASUMI-1 (40% & 60% less cells vs control vector) & exhibited a sensitizing effect to cytarabine (ara-C) treatment (22% & 10% higher cell reduction vs control vector + ara-C). Treatment with PTX showed upregulation of miR-9 in KG1a & KASUMI-1 (520% & 220%) & subsequent reduction of ERG mRNA (50% & 70%) & protein levels (10% & 20%). A sequential treatment with PTX 6h previous to ara-C led to higher amount of cell reduction compared to simultaneous administration indicating a sensitizing effect of PTX to standard chemotherapy. Pri-miR-9 expression was significantly down-regulated in BM of AML pts vs healthy individual BM (P < .01). While low miR-9 expressers showed promoter hypermethylation (P = 0.003) the methylation of high miR-9 expressers & healthy individuals was comparable (P = 0.66). High pri-miR-9 expressing AML pts had lower cumulative incidence of relapse (P = 0.04) & longer overall survival (P = 0.03). miR-9 directly regulates ERG expression & high expression associates with improved survival in AML. Increasing miR-9 levels with PTX might enhance the response to chemotherapy & improve outcome of AML pts. Disclosure: No conflict of interest disclosed.

Oncol Res Treat 2016;39(suppl 3):1–327

327

CONTENTS AUTHOR INDEX

Author Index Oncol Res Treat 2016;39(suppl 3):329–348

A Abd Kadi S.S.S. V485 Abdulla D. P300, P568, V134 Abendroth A. V997 Abenhardt W. P283, P602, V394, V693 Abken H. V1018 Abolfathi M. P217 Abreu D.R. V676 Abu-Jawad J. V997 Acker F. V715 Ackermann A. P919 Acker-Palmer A. P288 Adamski J. V716 Adès L. P517 Adjallé R. V45 Advani A.S. V658 Agis H. P235, P240 Ahlborn M. V792, V1022 Ahmadova G. V769 Ahmed F. P199 Ahn J. P578 Aigner A. P301, P312, P873 Aigner M. P895 Ajib S. P558, V1015 Akyüz N. V146, V985 Alakel N. V180, V756 Al-Ali H.K. P561, V29, V106, V178, V403, V467, V770 Alashkar F. P222 Alavi S. P884 Al-Baldawi Y. V717 Al-Batran S.-E. P864, V1023 Alberich-Jorda M. V660, V831 Albers C. V723, V787 Albers N. V358 Albert B.V. P202 Albrecht S. P219 Alchalby H. V30, V773 Aldaoud A. P855, P864, V1026 Alghamdi M. P513 Alghisi E. V405 Alimov A. P912 Al-Maarri M. V59, V717 Almishari M. P209 Alsadeq A. V659 Al-Sawaf O. P855, V685 Alsdorf W.H. V687 Alshehri N. P209 Alsohaibani L. P209, P513 Alt A. P287, P903 Alt J. P287 Altahan R. P209, P513 Altdoerfer V. P285 Alt-Epping B. V376, V792 Althaus K. P904 Althoff S. V1017 Altmann T. P280, V464 Amann E. V667 Ambarkhane S. P246

Accessible online at: www.karger.com/ort

Ambrosch P. P573 Amillano K. P597 Amler S. V356 Andrade-Navarro M.A. V840 Andre M. V465 Andrea M. V665 Andreß S. V764 Anell Rendon D. P894 Angelillo-Scherrer A. V85 Anhuf J. P212 Antes G. V123 Antlanger M. P240 Antonia S.J. V135 Apanovich N.V. P877 Apostolou P. P902 Appel T. P962 Araghi M. V669 Aranda Lopez P.C. P274 Aranyossy T. V485 Arasteh K. P256 Arat P. P537 Arlt W. P248 Armbruster N. P954 Armstrong S.A. V349 Arndt V. P885, V132 Arnold R. P232, P266, P282, P564, P850, V483, V1011, V1017 Arock M. V103 Arriaga Y.E. P304 Arrieta O. V135 Arsenic R. P593 Arteaga M.-F. V987 Asadi K. P282 Aschauer G. V358 Ascherfeld L. V796 Asemissen A.M. P281, V985 Assenov Y. V809 Aßmann M. P566 Assouline S. V1007 Atunes E. V667 Augsberger C. P280, V464 Augstein P. V339 Aul C. P935, V313, V738 Aulitzky W.E. P525, P546 Aulwurm S. V354 Aumann K. V143 Aurich K. P904 Austein T. P255, P615 Autenrieth S.E. P954 Avigdor A. V141 Ayuk F.A. P234, P934, V30, V45, V773, V485 Azeh I. P212, P530 Azrilevich A. P888 B Baccarani M. P201 Bach C. P229 Bacher U. P218, V403, V404, V779, V781 Bachhuber P. P511

Bacigalupo A. V1015 Backert L. P551, V145 Badbaran A. V485 Baerlocher G.M. P201, V155, V725, V728 Baertsch M.-A. P908 Bahlo J. P855, P860, V33, V395, V685 Bahra M. P862, P866 Baier J. P537 Baig M. V1028 Bairey O. V34 Bajrami Saipi M. P507 Bakchoul T. P514 Balasubramanian G.P. V78 Balasubramanian S. V141 Baldauf M. V157 Baldus C.D. P281, P578, V32, V347, V358, V401, V483 Balermpas P. P566, V339 Ballinger M. V344 Balzer H. P229 Bamberg M. V354 Bamopoulos S.A. P526 Bangerter M. V771 Bankstahl U.S. V1023 Baraldi A. P239 Barcellini W. V476 Bargou R.C. P231 Barile M. V639 Barlesi F. V135 Barr P.M. V34 Barriuso J. V670 Barth J. V390 Bartlett N.L. V34 Bartolomäus A. P872 Basara N. V355, V358 Bäsecke J. V404 Baselga J. P597 Bashari M.H. V387 Bassermann F. V718, V729 Bastian L. V347 Bastian S. P534 Basu O. P557 Batereau C. P963 Bauer R. V838 Baues C. V342 Baum V. P289 Baumann F.T. V51 Baumann U. P956, V718 Baumann W. P603, V172 Baumgardt H. P615 Bäuml E. P583 Bechmann I. P312 Beck J. P848, V665 Becker C. P204, P205, P863 Becker H. V182 Becker H.J. P893 Becker K. P198, P938 Becker K.-P. P605 Becker S. P282

CONTENTS AUTHOR INDEX

Beckert J. P508 Beelen D.W. P557, P559, V1014 Behlendorf T. P962 Behre G. P190, P495, P559, V348, V481, V660, V783, V831, V834, V1012, V1013, S. 327 Behrenbeck F. V483 Behrend M. P938 Behringer J. P214 Beier C.P. P926 Beier D. P926 Beier F. P229, P563, P573, P926 Beier M. P581 Belanger B. P863 Belka C. P566 Belleville E. P212 Bellos F. P520 Ben Batalla I. P604, V838 Benedikter J. V134 Benkisser-Petersen M. V60 Bennett B. P879, V341 Bennett J.M. V738 Benser J. P603 Bentz M. P965 Berdel W.E. P194, P197, P198, P536, V32, V181, V840, V987 Berenstein R. V483, V731, V732 Berg T. P558, V1015 Bergelt C. V796 Berger A. V1018 Berger D. P601, V668 Berger K. P522, P523 Berghorn E. P879 Bergmann A. V786 Bergmann L. P880, P881, P882 Bergmann M. V33, V685 Bergmann S. V756 Bergmann T. P566 Bergmann U. V783 Bergwelt M. P860 Berking S. P261 Berlin C. V1020 Berner J. V386 Bernhard R. V88, V89 Berry S. P879 Bertoni F. P527 Bertram M. P603, P938, V394 Bertz H. P556, V173, V801 Bethge W.A. P263, P264, P271, P551, P552, P553, P559, P560, V354 Betz B. V484 Beuschlein F. P248 Beutel G. V678, V755 Beutner D. P566 Bewarder M. P245, V720, V734 Beyer F. V403 Bezhanova S. V717 Bianchi P. V476 Bianconi D. P890 Biehl L.M. V44 Biersack H. V358 Bigalke I. V464 Bileck A. P853 Bilich T. V683

Author Index

Bill M. P190, P561, V348, V481, V660, V665, V783, V831, V1012, V1013, S. 327 Billing C. V90 Bilmes R. V158 Bilotti E. V391 Binder C. P595 Binder M. V146, V442, V985, V996 Binninger A. V986 Birghan C. V632 Birkmann J. V75 Birndt S. V474 Bischofberger I. S. 322 Bischoff H. P287 Bischoff M. P237, V170, V175 Bischoff S. P862, P866 Bisht S. P229 Bittenbring J.T. P229, V715 Bittrich M. P545 Blagitko-Dorfs N. V350 Bläker H. P862 Blanc J.-F. P863, P865 Blankenstein T. P284, V1017 Bläß M.F. V345 Blatt K. V668 Blau I.W. P230, P232, P233, P235, P281, P564, V483, V731, V732 Blau O. P230, P232, P233, P564, V483, V731, V732 Blay J.-Y. V990 Blazek T. P539 Bleckmann A. P595 Bleckmann D. V60 Bleeke M. P919 Blessing F. P515 Bleul S. V182 Bloch W. V51 Blum H. V32, V347, V686 Blum K.A. P246 Blum P. V664 Blumenstengel K. P530, P856, V390 Blumenstock G. P506, P507 Boccadoro M. P239 Boch T. P219, V92, V401, V686 Bochtler T. P217, V358 Bock C. P852 Bock T. V842 Böck S. V367 Bode S. P260 Boden K. V674 Boehm A. V103, V792 Boese S. V101 Bogatyrova O. V809 Bogdanova A. V476 Bögeholz J. P265, P613 Bogner C. P854 Bohlander S.K. V356 Bohle R.-M. V720 Böhm A. V442 Böhme C. V90 Böhmer F.-D. P202 Bohn J.-P. P858 Bohnenberger H. V719 Bohnet S. P287 Boix O. P941, P942 Bojunga J. P920, P921, P922

Bokemeyer C.

P604, P933, V146, V651, V687, V763, V796, V838, V985, V996 Bolder U. P311 Bolenz C. V1032 Bolkun L. V392 Böll B. V454 Boller E. V385 Bommer M. P517, P600, V475 Bommhardt U. P847 Bonekamp D. P600 Bongartz H. P510 Bonifacio E. P516 Bonifacio L. P190, V348 Bönig H. V666 Bono P. P888 Bonzheim I. P187 Bootz F. V340 Boquoi A. P537 Borchert K. P582, P876 Borchmann P. V112, V160 Borchmann S. P253 Borges jr. U. V172 Borggrefe J. V342 Borghaei H. V135 Borkhardt A. V78 Bornfeld N. P614 Bornhäuser M. P194, P258, P498, P516, P559, V180, V181, V358, V402, V694, V756, V780 Borozdin W. V759 Börries M. P913 Borte M. P956 Bose T. P847 Bosse R. P906 Böttcher M. P188, P229, V144 Böttcher S. V395 Botteman M. P865 Böttner A. V780 Bouabdallah K. V1007 Bouillon A.-S. P563, P573, P926 Boxhammer R. P235 Bradley A. V833 Bradner J.E. V349 Bradwell J. V662 Braess J. P197, V356, V474 Braga E.A. P877 Brägelmann J. V340 Brahmer J.R. V135 Braicu E. P593 Braig F. V996 Bramhoff A. P515 Brandes V. P300, P568, P569, V134, V676, V843, V988 Brandt A. V985 Brandt J. P528 Brandt K. P258 Brandts C. V170 Brass V. P913 Bräuer-Hartmann D. P495, V660, V831, V834 Braulke F. V779 Braun M. V388 Braun S. V756 Bräundl K. V356 Brauneck F. P280 Brauner M. V759

Oncol Res Treat 2016;39(suppl 3):1–348

330

CONTENTS AUTHOR INDEX

Breccia M. V770 Brecht P. P309 Breese E.H. P497 Brendel C.A. V91, V156 Brenner B. V644 Brenner M.K. V466 Brettschneider J. P273 Breyer M. V477 Brinkmann F. P292, V384 Brioli A. P239, P542 Brittain D. V770 Brockhoff H. P236 Brocks D. V809 Brodhun M. V674 Broglio K. P879 Bron D. V1007 Brossart P. P193, P229, P257, P543, P925, V86, V108, V340 Brosteanu O. V178 Brück P. P875 Bruckner T. P908 Brueckner F. P269, P518, P609 Brüggemann M. P262, V77, V347, V393, V464, V480, V684, V975 Brümmendorf T.H. P194, P563, P573, P926, V134, V153, V155, V156, V358, V403, V675, V683, V1020, V779 Brummer T. P913 Brunkhorst F.M. V474 Bruns H. P229, V144 Buchheidt D. V92 Buchhold B. V795 Buchholz D. V763 Büchner T. P197, P198, V832 Buchtele N. V151 Bücklein V.L. P261, V464 Budach V. V339, V1000 Budczies J. P600 Buech T. P873 Bueckner U. P219 Buettner R. P288 Bug G. P558, V779, V1015 Bugl S. P503, V56 Bühring H.-J. V354, V465 Buisman S. V677 Bujor L.-V. V1014 Bullinger L. P292, V810, V830, V1021 Bulycheva E. V402, V686, V784 Bunjes D.W. V93, V1011 Burchert A. V155, V156, V355, V493, V675 Burger J. V34 Burkhard O. P214, P868 Burmeister T. V357 Burmeister Y. V659 Busch C.-J. V996 Busch H. P913 Busch K. V639 Busch L. P229 Busch M. P203 Busemann C. P514, V795 Buske C. P246, P292, V183, V391, V392, V411, V830, V835, V971 Buß E. P210 Busse A. P281

331

Busse M. Büttner B. Büttner M. Büttner R. Buxhofer-Ausch V. Byrd J. Bystry V. Bystrykh L. C Cabron A.-S. Caca K. Calvo E. Cameron S. Campo E. Candido S. Canepa L. Cantor H. Capper D. Carcereny E. Cardone M.H. Cario G. Cario H. Carles J. Castaños-Vélez E. Castillo J.J. Catalano L. Cavo M. Cebulla S. Cella D. Chakupurakal G. Chamnitzer O. Chan E. Chanan-Khan A. Chase A. Chatterjee M. Chatterjee S. Chawla S. Chayahara N. Chemnitz J. Chen C.-W. Chen L.-T. Cheng M. Childs B.H. Chitic A.P. Chmielewski M. Chortis V. Christian A. Christopeit M. Christopoulos P. Chudziak D. Ciardello F. Clark K. Clark R. Claus R. Clausen J. Cleary M.L. Clemens J. Clement J.H. Clow F. Cockwell P. Cogliatti S.

Oncol Res Treat 2016;39(suppl 3):1–348

P937 V180 P229 P286, P300, P568, P569, V59, V134, V676, V717, V843, V988 V442 V809 P262 V677 P249 V644 P888 P924 P525 V842 P239 P257 V78 V676 V387 V659 V396 P887, V1031 V840 V391 P239 P239 P590 P544, P879 P307, P857, P951 V1024 P888 V141 V345 P231, P235 P288 V990 V998 P267 V349 P863, P865 V34, V392 V1007 P311 V1018 P248 P581 P495, P559, V30, V45, V485, V773 P289 V417 P304 P954 P195 P913, V60, V143, V182 V442 P497 V768 P204, P205, P206, P899, P900, P911 V34 V662 P534

Colomer D. Conradi I.S. Cook G. Cook M. Corbacioglu S. Cordes S. Cornell R.F. Cornely O.A. Corral J.J. Corso J. Cortelezzi A. Cortes J. Cortes J.E. Coutre S. Craig A.R. Craig M.D. Cramer P. Cremer M. Crinò L. Crivello P. Croner R. Cross M. Cross N.C.P. Crown J. Crysandt M. Csanadi A. Cubas Córdova M. Cubillo A. Cunningham D. Cusan M. Cygon F. D D´abundo D. D'Adamo D.R. Daga S. Dahlenburg J. Daisne J.-F. Dammann E. Damnali G. Danielczok J. Dannenmann B. Daria D. Darzentas N. Dastani H. Davis C. Davis Z. De Braud F. De Haan G. De Jong F. De la Pena L. De Stanchina E. De Wit M. Dean E. Deangelo D.J. Debatin K.-M. Debole J. Decker J. Decker S. Decker T. Deiser K. Delacruz A. Delgado J.A.

Author Index

P525 P296 P544 V662 V398 V438, V439 V391 V42, V44 V676 V719 V476 P597 V158 V34 P195 P195 V141, V395, V685 P528, P532 V135 V1014 P924 P190, V90, V178, V348, V481, V727, V1012, S. 327 P207, P517, V105, V169, V345, V682, V768, V769, V726 V343 P563, V989 P913 P604, V838 P863, P865 V1007 V349 V643, V989 V1017 V990 V482 P617 V644 V678 P940, S. 324 V476 V88 V830 P262 P879, V341 P544 P852 P888 V677 P863, P865 P597 V349 P287, V358, V675 V670 V658 V655, V656 V1033 V759 V143, V733 P250, V693, V1022 P280, V464 V1030 V673

CONTENTS AUTHOR INDEX

Demetri G.D. V990 Demir S. V655, V656 Demirkan F. V141 Demirtas D. V669, V790 Demmer P. P913 Demonty G. P302, P303 Demut J. P900 Demuth I. S. 326 Dengler J. P210, P855, V154, V156, V675, V728 Denkert C. P604, P862, V840 Dennert G. V95 Denny M. P274 Denzlinger C. P870, P871 Depenbrock H. V131 Depenbusch R. P602 Depré F. V479 Derigs H.-G. P195, V648 Derigs P. V469 DeRosa M. P879, V341 Derudas D. P239 Desai K. V135 Deschler B. V123 Deshpande A.J. V349 Desole M. P910 Dethmers B. V677 Detlefsen N. V390 Deubrecht S. P957 Devereux S. V34 Dhawan A. P258 Di Christanziano V. P267 Di Cosimo S. P597 Dick T. V348 Dickerhoff R. V397 Dickmann J.R.M. P255 Diedrich D. V50 Diefenbach K. P941, P942 Dieing A. P287 Diem M. V634 Dienst A. V484 Dierks C. V143, V733, V785 Dierks S. P218, V404, V781, V782 Dietel M. P593 Dieterich M. P594 Dietrich D. V340 Dietrich S. P262, V61, V1010 Dietz A. P572, V35, V92 Dietz C.T. P201, V156, V728 Dietzfelbinger H. P224 Dilhuydy M.-S. V141 Dimopoulos M.A. V391, V392, V730 Dirkmann D. V477 Dirksen U.T. V322, V381 Dirnhofer S. P496 Distel L. V1000 Ditschkowski M. P557, V355, V1014 Dittmar G. V439 Dittmer U. V1014 Dittrich T. V661 Diwoky S. P548 Doan J. P879 Dobrosch L. P543 Doebele C. V719 Doehn C. P883 Doerfel S. P864 Doerken B. P230, P232, P233, P866, V483, V731, V732

Author Index

Dohle I. P860 Döhner H. P292, V93, V183, V358, V771, V809, V830, V835, V1021 Döhner K. V183, V680, V771, V781, V830, V835 Dold S.M. P538, V123, V801 Doll S. V832 Döpper J. P258 Dörfel D. P285, P551, V354 Dörfel S. V385 Döring P. P969 Dörken B. P266, P282, P284, P850, V357, V789, V841, V1017 Dormann C. P213 Dorn-Beineke A. V638 Dörr J.R. V841 Douillard J.-Y. P302, P303 Dreger P. P262, V29, V61, V93, V355, V437, V466, V1010 Dreischer T. S. 325 Dresel A. P944 Dressler S. P590 Dreyling M.H. P524, P526, P531, V685, V1007 Driessen C. V361, V806 Drolle H. P583, P940 Drube J. P202 Drudge-Coates L. V1030 Du J. V789 Du Y. P848 Duensing S. P884, P886 Duerr-Stoerzer S. P263, P271, P279 Dührsen U. P222, P223, P859, V358, V430, V475, V477, V780, V786 Dukal H. V686 Duran Graeff L. V42 Dürig J. P530, P859, V786 Düring U. P582 Dürkop J. P599 Dürst M. V823 Dutcus C.E. V995, V998, V1028 Duyster J. P227, P228, P242, P512, P538, P891, P906, P913, P939, P943, V60, V87, V123, V143, V182, V327, V723, V733, V785, V787, V801, V833 Dworzak M.N. V634 Dyshlovoy S. P919, V687 E Eberhardt J. Eberhardt W.E.E. Eberlein C. Ebert A. Ebner F. Echchannaoui H. Eckert R. Edelmann T. Edelmann-Stephan B. Eder M. Eder T. Edmaier K. Egerer G. Eggers H. Eggert A. Eggert J.

V107, V772 P289, P295, V135 V181 P936 V764 P273, P275, V667 V675 V643, V665 P509, P510 V678 V1000 V183 V43, V663, V989 V1029 V78 V390

Ehleiter H. Ehninger G.

P535, P547 P194, V32, V180, V181, V358, V756, V780, V1025 Ehrenfeld S. P242, P508 Ehscheidt P. P214, P868 Eichhorst B. P855, P860, V33, V395, V685 Eichler H. P522 Eichner R. V729 Eigendorff E. P517, P611, V153, V155, V156, V989 Einsele H. P231, P235, P237, P272, P545, P546, P606, V75, V360, V666, V728, V729, V801, V1008, V1011 Eisenwort G. V103 El-Khouly F. V670 Elsel W. P306 Elsner M. P206 Emde T.-O. V986 Emig M. P875 Emmert S. P575 Enard W. V406, V839 Endell J. P246 Endris V. V1000 Engel E. P938 Engel N. P261 Engelhardt M. P227, P228, P235, P237, P538, P546, P547, P891, P906, P939, P943, V44, V87, V123, V801 Engelke A. V395 Engel-Riedel W. P289 Engert A. P253 Engert R. P944 Enokida T. V998 Erben P. V1032 Erbes T. P913 Erdmann-Schneider P. P578 Erhart D. P903 Erichson A. P961 Ernst J. P203 Ernst M. P848 Ernst T. P202, P203, P207, P910, V31, V157, V179, V345, V346, V474, V726 Escherich G. V763 Escudier B. P878 Escuriola-Ettinghausen C. P522 Eshraghi P. V183 Estenfelder S. P243 Ethell M.E. P517 Eugster A. P516 Evans T.R. V670 Ewerth D. V87 Exner A.-K. V52 Eyb V. P553

Oncol Res Treat 2016;39(suppl 3):1–348

332

CONTENTS AUTHOR INDEX

F Fabarius A.C.

P201 P216, V104, V105, V686, V728, V768, V769 Fabisch C. V155 Fabri M. V342 Fadle N. P245, V720 Fahldieck C. V107 Falcone A. P304 Faldum A. V356 Falge C. V728 Falk M. P296, V133 Fan F. V387 Färber J. V44 Farlik M. P852 Farnoud N.R. V349 Farshi P. P558 Farsijani N.M. P223 Fasan A. V637, V657 Fasching P.A. P604 Fasshauer M. P956 Fassnacht M. P248 Fassunke J. V988 Fatho M. P273 Faude O. V51 Faul C. P263, P264, P271, P551, P552, P553 Faust M. P555 Fauth F.W.B. V644 Fayette J. V135 Feder K. V183 Fehm T. P604, V388 Fehrenbacher L. V344 Fehse B. P234, V485 Feiten S. P214, P307, P857, P868, P951, P967 Felices M. P918 Felip E. V135, V676 Fellas G. P244, P252 Felthaus J. P891, V87 Fenchel K. P283 Feng Y. P589 Feng Z. V349 Fenk R. P537 Fermo E. V476 Fernández de Larrea C. V391 Fernández-Sáiz V. V729 Ferrara F. P239 Ferreira M.V. P563, P926 Ferry D. P875 Ferstl B. P235 Fette G. P545 Feuerbach M. P550, P861 Feuring-Buske M. V183, V830, V835 Fey M. V355 Fey S. V686 Feyerabend T. V639 Fichtner I. V840 Ficker J.H. V134 Fiechtner N. P561 Fiedler E. V101 Fiedler M. V1014 Fiegl M. V356 Fiesel P. V78 Fietkau R. P566, P573 Fietz T. P212, P950, P952, V394 Fifer L. V662

333

Finck A. V355 Fink A.-M. P855, V33, V395, V685 Fink G. P906 Finke J. P556, V355 Finkernagel F. V156 Finzsch M. P566 Fischer B. P933 Fischer D. P602 Fischer J. P287, P515 Fischer K. P855, V33, V395, V685 Fischer M. P899, V94 Fischer R.N. P290, P300, P568, P569, V134, V676, V843, V988 Fischer T. P509, P510, P546, P847, P917, P957, V29, V44, V107, S. 326 Fischer von Weikersthal L. P530, V1022 Fischer-Maranta A. P534 Fishman M. V1028 Flach J. P218, V404 Fleischhauer K. V1014 Fleischmann M. P196 Fleitz A. V171 Flieser-Hartl M. P583 Flörcken A. P284, P850 Florin A. P286, P288 Florschütz A. P617, V29, V643 Flossdorf M. V639 Foerster K. V60 Föhring D. P859 Follmann M. V175 Follo M. P228, P512, P867, V327, V723, V733 Folprecht G. V1025 Foltz L. V770 Ford C. V146, V985 Forget F. P303 Forstbauer H. V643 Förster S. P570 Francke S. P258 Franke G.-N. P190, P561, V106, V348, V481, V727, V783, V1012, V1013 Fransecky L.R. V32, V483 Fraser G. V141 Frech M. P592 Freigang F. P309, P550, P861 Freitag A. V78 Freitag J. P916, V140 Freitag S. P268, P269, P518, P575, P607, P609, P961, P964, V50, V999 Frenzel L.P. V717 Frerich B. V999 Freund A. P898 Freund M. P269, P932 Freunek G. V156, V675 Frey B.M. P259 Frey M. V672 Fricke S. P495, V831 Frickhofen N. P546, V474 Friday D. P521 Fridrik M.A. P221, P966 Friedrich K.-H. P205 Friedrich M. V761 Friedrich S. P885, V132 Friesen C. P903

Oncol Res Treat 2016;39(suppl 3):1–348

Friesenhahn V. Frille A. Fritsch K. Fritsch R. Fritsch S. Fritz C. Fritz L. Fröhlich M. Fröhling K.-P. Fröhling S. Frömberg A. Frosinski J. Frost N. Fröstl C. Fruth A. Fuchs A.R. Fuchs D. Fuchs N. Fuchst C.S. Fuellen G. Fujino K. Füllgrabe M. Fürst D. Fuxius S. G Gaber T. Gaidano G. Gaksch L. Gale R.P. Galfetti E. Galic M. Gallego J. Galvan P. Gamper E.-M. Gand C. Ganser A. Ganser M. Ganster C.

P307, P857, P951 P298, P567 P867 P867, P913 P261 V717 V91 P600 P571 P600, V754 P301 P959 P949 P581 V469 V56, V58 P221, P966, V724 V477 P875 P848 V998 V684 V1011 V1022

P190 P246, V34 V482 V774, V812 P613 P283 P302 P597 V124 P249 V678, V779, V1029 P225, V56 P218, V403, V404, V781, V782 Garcia Marquez M. P893 Garcia-Carbonero R. P304 Garcia-Marquez M. V342 Garcia-Sanz R. V391, V392 Garcia-Vargas J. V1007 Garlipp B. V643 Garon E. P293 Garz A.-K. V406, V729 Gassner C. P259 Gastl G. V157, V724 Gattermann N. P248, V153, V313, V403 Gaudio E. P527 Gauler T.C. V997 Gawlitza A. V346 Geer T. P511 Geier B. V1022 Geismar D. V323 Geissler S. V724 Gelbenegger G. V151 Gellert K. P924 Gelß P. V686 Gencer D. P605 Gensch V. P604, V838 Gentner E. V835 Geörg C. P600 George S. P878 Georgi J.-A. V181

Author Index

CONTENTS AUTHOR INDEX

Gerhardt A. Gerigk M. Gerisch M. Gerl A. Gerlach L. Gerloff D. Germing U.

P530, V841 V988 P941, P942 V1033 P294 P494, P495, P501, P502, V660, V831, V834, S. 327 P220, V313, V403, V404, V484, V697, V729, V738, V779, V781, V813 Gern U. V466 Gerner C. P853, P914 Gerngroß C. P927 Gerrard G. V726 Gerrlich C. P537 Gerth U. P197, P198, P536 Geßner D. P957 Geue K. V761 Geyh S. V484 Gezer D. P229 Ghadjar P. P866 Ghia P. V34 Ghibelli L. P914 Ghosh J. V770 Giagounidis A. V313, V355, V779 Gianoukakis A.G. V995 Giers G. P515 Gieseler F. P898 Giessen-Jung C. V647 Giles F. V153, V157, V724 Gillessen S. P887, V1031 Giraldo P. V770 Gisslinger H. P240 Gjertsen B.-T. V157 Glade J. P917 Glaeser D. P964, V791 Glaeser H. P964 Glag S. P968 Gläser D. P269, P579, P584, P594, P872, P961, V665, V999 Gläser H. P269, P585, V793 Glass Ä. P268 Glaß B. V403, V404, V781 Gleich S. P934 Gleiss A. P890 Gleixner K.V. V668 Glitsch A. P969 Gloeckner C. P291 Glossmann J.-P. P237, P290, V170, V175, V988 Glubokova E. P874 Gockel I. P312, V1024, V1027 Goebeler M.-E. V156, V675 Goebell P.J. P880, P881, P882, P883 Goecke T.W. S. 326 Goede V. V33 Goerner M. V390 Goetz M. V1021 Gogishvili T. V1016 Göhler T. P305, P308, P310, P566, P602, P856, P880, P881, P882, V643 Göhring G. V105 Gökbuget N. V347, V480, V658, V684, V1015 Gökkurt E. V985 Gold R. P956

Author Index

Goldschmidt H.

P235, P237, P238, P540, P544, P908, V663, V800, V801, S. 322 Golfmann K. P286, V389 Gollhofer A. V173 Gonzales R. P597 Göpel W. P851 Gorantla S.P. V723 Gorbatchevsky I. V1007 Görlich D. P197, P198, P536, V356 Görner M. V358 Göttel R. P297, P856 Götz M. V93 Götze K.S. V355, V358, V406, V729, V688, V779, V839 Götze T.-O. P864 Götzel K. V1024 Gourri E. P259 Goy A. P246, V141 Grab A.L. V673 Grabarczyk P. P249 Graef T. V391, V392 Graf A. V32, V347 Graf E. V327 Graf T. V764 Gräfe C. P900, P911, V94 Gralla R.J. V341 Gramatzki M. P235, V659, V671, V1011 Granell M. V391 Grassinger J. P608, P613 Grath S. V406 Gratt J. P887, V1031 Grebhardt S. P598, P950 Greger N. V50 Greif P.A. V32, V347 Greil C. P891, P906 Greil R. P213 Greinacher A. P514, P904 Greiner J. V93, V358, V1021 Greinix H. P934, V762 Greul R. P221 Greve G. V350 Greystoke A. V670 Griesinger F. P287, P295, P296, V133, V343 Griesshammer M. P519, V451, V770, V771 Grignani G. V990 Grigoleit G.-U. V666, V688 Grimm J. P190, V348, V481, V783, V1012, V1013, S. 327 Grischke E.-M. V693 Grob T. V996 Grobe N. P250 Grohé C. P289, V343, V676 Gröschel S. P600 Gröschl B. P511 Grosicki S. V34, V141 Grosse-Hovest L. V354, V465, V1019 Große-Thie C. P268, P269, P518, P575, P579, P585, P588, P589, P607, P609, P610, P964, V478, V791, V793, V999 Grothey A. P304 Grugel R. V394 Gruhn B. P203 Grüllich C. P883, P886 Grundeis M. P306

Grundheber M. Grünebach F. Grunert J. Grunewald R. Grünewald V. Grünwald V. Gu L. Guan X. Gudzuhn A. Gueller S. Guilhot J. Gundel D. Güngör T. Günter M. Guo M. Gupta M. Gupta V. Guth D. Guthoff R. Gütz S. H Haack K. Haack L. Haag C. Haalck T. Haap M. Haas A. Haas C. Haas E.-M. Haas R. Haase D. Haase J. Haase M. Haastert B. Habra M.A. Habringer S. Hacker U. Hackstein H. Hadaschik B. Hadzijusufovic E. Haeberle L. Haegeman G. Haehnel S. Haen S.P. Haertel N. Haferlach C.

Haferlach T.

Haffner F.T. Haffner I. Hafner F.T. Hagedorn S. Hagmaier V. Hähling D. Hahn D. Hahn M. Hahn-Ast C.

Oncol Res Treat 2016;39(suppl 3):1–348

P214 P285 V1007 P210 P572 P878, P879, P883, V337, V1029 V809 P303 P969 V1015 V156, V675 P606 P608 P954 V990 V730 V770 P953 P548 P289 V664 P584 V984 V985 P503 P945 P898 P238 P217, P248, P537, V484, V779, V813 P218, V403, V404, V779, V781, V782 P204 P248 P220 P248 P927, V688 V647 V91, V766 P886 V668 P212 P914 P312 P264, P285, P551, P552, P553, V465 P864 P191, P201, P520, P965, V105, V142, V345, V353, V637, V657, V686, V769 P191, P520, V104, V142, V168, V353, V630, V637, V657, V698, V769 P941 V1022 P942 P884 P271 V29 P566 V61 P543

334

CONTENTS AUTHOR INDEX

Haier J. Hain S.-A. Haitel A. Hajek R. Hallas C. Hallek M.

P933 V61 V336 V392 P296, V133 P197, P288, P290, P855, P860, V33, V59, V141, V395, V685 Haltaufderheide J. V982 Halter J. P934 Hamel T. P292 Hamm D. P869, V1026 Hammers C.M. P859 Hammers H.J. P878 Hammersen J. P204, V46 Handgretinger R. P551, V710 Hänel M. P530, V178, V358, V728 Hänggi P. V476 Hänig J. P213 Hannig C. P241, V643, V644 Hansen R. P212, P214, P598 Hansmann M.-L. V720 Hapke G. P530 Harbeck N. P597 Harbison C. P888 Harde J. P598 Harder L. V404 Harich H.-D. P880, P881, P882 Häring M.- F. P560 Hartmann A. V1032 Hartmann A.-K. P274 Hartmann B. V759 Hartmann J.-U. P495, V660, V831, V834 Hartmann S. V720 Hartmann W. V987 Hartung S. P892, P901 Hartung T. V32 Hartwig U.F. V672, V1018 Hasenbank C. P573 Hasenclever D. V642 Hasford J. P201, V155, V725 Hass H.G. P601, P870, P871 Hastka J. P605, P612 Hauenstein K. V478 Haug N. P505 Hauptrock B. P554 Hausmann A. P261, P559 Hausmann D. V768 Hausschild J. V687 Häusser L. P186 Haustermans K. V644 Hautmann M.G. V1000 Haverkamp C. P943 Haverkamp T. P215 Healey D. V135 Hebart H. P219, P546, V771 Hecht A. P555, V832 Hecht J. V347 Hecht M. P566 Heemskerk M.H.M. V464 Heffner L.T. V391 Hegenbart U. V29, V61, V661, V664 Hegewisch-Becker S. P869, P938, V174, V1026 Hehlmann R. P201, V155, V156, V725, V728

335

Heidel F.H. Heidel K. Heidemann E. Heidenreich A. Heidenreich D. Heidenreich F. Heider A. Heider M. Heidrich K. Heidsieck T. Heil G. Heilmann V. Heilmeier B. Heim D. Heimann A. Heine A. Heine M. Heine S. Heinemann V. Heinicke T. Heining C. Heinrich B. Heinrich D. Heinrichs S. Heinz F. Heißner K. Heitmann J.S. Heitzer E. Helas S. Heller S. Hellmann A. Hellmann M. Hellmich M. Hellmich U. Hemmati P. Hempel D. Heni M. Henke D. Henke R.-P. Henkenius K. Hennig L. Henrich F. Henschler R. Hensel M. Hentrich M. Hentschel L. Henze L.

Henzler T. Herac M. Herbig A. Herbst R. Herda S. Herget G. Herhaus P. Herich-Terhürne D. Hering-Schubert C. Herling M. Herlyn P. Hermann B. Hermann E. Hermann S. Hermes-Moll K. Herndlhofer S.

Oncol Res Treat 2016;39(suppl 3):1–348

P847, P917, V107, V771, V772 P498 V384 P887 P555 P516 V792 V729 P258 V1023 P215 P953 P197 V155 V1017 P925, V86 P255 P525 V647 V29 P600 P511 P887, V1031 V477 P518 P503, V989 V56, V58 V482 V780 P577 V34 V135 P569 P221 P232, P266, P564, V762 P602 P553 V384 P296 V91 P312 P895 P523 P530, P956 P546, V44, V1033 P916 P268, P269, P518, P575, P585, P588, P589, P607, P609, P610, V478, V793 V768 P890 V1034 V156, V178, V675 V1017 P228 P927, V688 P222, V475 P305, P308, P310 P267 P610 V674 V1016 P885, V132 P603 P213

Herold S. V181, V780 Herr W. P200, P914, V468, V672, V753 Herrmann D. P262, V393 Herrmann E. P880, P881, P882 Herrmann H. V103 Hertenstein B. P546, V390 Hertz L. V476 Herzberg P.Y. V762 Herzog A. V393 Heß A.-K. V339 Heß G. P531, P546, V65 Hessenkemper W. P510 Heßling J. P530 Hettmer S. P913 Hetzel M. P292 Heuckmann J.M. P291, V343 Heukamp L.C. P288, P291, V133, V343 Heuser M. P195, V358, V737, V803 Heußner P. V138, V760, V762, V794, V979, V980, V981 Heydenreich M. V49, V53 Heydrich B. V728 Heydt C. V843, V988 Heymanns J. P307, P857, P951, P967 Heyn C. V484 Heyn S. V29, V665 Heyne N. V662 Hiddemann W. P197, P261, P280, P524, P526, V138, V183, V356, V393, V464, V760, V794, V973, V979, V980, V981 Hielscher C. P602 Hiemer S. P617 Hiemeyer F. V1007 Hildebrandt B. V484, V643 Hildebrandt G. V999 Hildenbeutel S. P906 Hilgarth M. V669, V790 Hilgendorf I. P196, P269, P611, P892, P909, P910, P934, V46, V50, V441, V762 Hilger N. P495, V831 Hillengass J. P238, V663 Hillmen P. V34 Hilser T. V997 Hindahl H. P311, V780 Hinke A. V390 Hinterleitner C. P503 Hipp J. P867 Hipper A. P295 Hirsch F.R. V131 Hirt C. P249, V29 Hlawa W. V791 Ho A.D. P262, P528, P532, P907, P908, V43, V61, V93, V437, V466, V661, V663, V664, V1010 Hochhaus A. P196, P201, P202, P203, P204, P205, P206, P207, P239, P517, P542, P611, P851, P892, P899, P900, P901, P909, P910, P911, P934, P959, V29, V31, V46, V94, V153, V155, V156, V157, V158, V179, V345, V346, V358, V370, V415, V417, V441, V474, V674, V675, V724, V725, V726, V728, V762, V771 Höchsmann B. V625 Höckendorf U. P499

Author Index

CONTENTS AUTHOR INDEX

Hoeffkes H.-G. V643 Hoelzer D. V347 Hoeper M.M. V678 Hoermann G. V103 Hofbauer L.C. V402, V784 Hofer S. V482 Höfer T. V639 Höffken G. V756 Höffkes H.-G. P530, P864 Hoffmann C. P576 Hoffmann C.M. V648 Hoffmann F.-A. V665 Hoffmann J.-M. V466 Hoffmann M. P958, V342 Hoffmann T. P573 Hoffmeister A. P312 Hofheinz R.D. P304, P864, P875, V644 V648 Höfler G. V482 Hofmann M. V354 Hofmann S. V93, V1021 Hofmann W.-K. P216, P219, P517, P555, P605, P612, V104, V105, V92, V401, V686 V728, V739, V768, V769, V779, V832 Hogge D. P195 Hohenfellner M. P886 Hohloch K. P534 Holderried T.A.W. P257 Hölig K. P516 Höllein A. V716 Holler E. P546, P559, V762 Höller K. P267 Hollerbach S. V644 Holms F. P604 Holstein K. P522 Holtick U. P267 Holtmann L. P948 Holzner B. V124 Honecker F. V687 Hopfer O. P244, P252 Hopfinger G. V685 Horger M. P533 Höring E. P525 Hörl R. V656 Horn L. V135 Horn M. V341 Horn P.A. V477 Horneber M. P590, V95 Hornemann B. P916 Horny H.-P. V104, V105, V768 Hörsch D. P920, P921, P922 Horst H.-A. P195, P576 Horstmann A. P519 Hose D. V673, V807 Hoshii T. V349 Hoster E. V393, V973 Howes A. V141 Hoya V. P614 Hozak R. P875, V131 Huber J. P944 Huber M. V156 Huber T. V723 Hubert K. V106, V178 Hubmann M. P224, P261 Hubmann R. V669, V790 Hubner R. P863, P865 Hübner A. P283

Author Index

Hübner J. Hübschmann D. Hückelhoven A. Hudecek M. Hug M.J. Hughes D. Hummel M. Hundemer M. Huober J. Huppa J.B. Hurtz H.-J. Huscher D. Hussung S. Huster A. Hutchison C. Huth A. Hutson T.E. Hüttelmaier S. Hutter B. Hutter G. Hüttmann A. Hutton E. Hutzschenreuter U. I Iżykowska K. Ihle M. Ihme S. Ihorst G. Illerhaus G. Illert A.L. Illmer T. Ilyas A. Imamura Y. Inagaki L. Innig G. Insa A. Inselmann S. Irger M. Isaakidis K. Istvanffy R. Ivanyi P. Ivics Z.

V377, V654 P600 V93, V466 P272, V1016 P227, P939, P943 P226 V339 P907, P908, V673 V764 P272 P530, P550, P861, P950 P956 P867 P560 V662 V44 V1028 P501, P502 P600 P524, P526 V475 V670 P297, P602 P249 V134, V843 V183, V830 P227, P538, V123, V801 P247, P566, V352 V87, V723, V733, V787, V833 P855, V153, V156, V675 P199 V998 V998 P297 V676 V156 P526 V32 V688, V839 V1029 V1016

J Jabbour E.J. V658 Jacob C. V178 Jacobasch L. P511 Jacobi A. P498 Jacobs L. V716, V729 Jacobs S. V793 Jaeger E. V138, V794, V979, V980, V981 Jaeger U. Jäger D. P600, P886, P888, V387 Jäger E. V1023 Jäger U. V669, V685, V788, V790 Jahn F. P549, P946, V96, V97, V461 Jähnke A. S. 322 Jahn-Kuch D. V28 Jahreis S. P892, P901 Jahwar M. V769 Jäkel N. V106, V178, V467 Jakob A. V358 Jakob C. V29 James D.F. V34 Jänicke M. P295, V174, V986

Janka G. V474 Jann J.C. V401, V686 Janni W. V114, V693, V764 Janning M. P604 Jansen J.H.M. P276 Jansen M. P960 Jansen-Winkeln B. P312 Janssen C. P299, V675 Janssen J. P211, P212, P305, P308, P310, V157, V724 Janssens A. V34, V141 Janssens J. V644 Janz M. P578 Janzen V. P543, P925, V675 Jarutat T. P235 Jaschonek K. V628 Jauch A. P217 Jawhar M. P555, V104, V105, V130, V768 Jedlickova Z. P558, V1015 Jehn C. P230, P266 Jenke R. P873 Jentsch-Ullrich K. V772 Jentzsch M. P190, P561, V348, V481, V665, V783, V1012, V1013, S. 327 Jenzen D. P961 Jeong J. P497 Jeremias I. V688 Jeromin S. P520, V142 Jia Y. V670 Jitschin R. V144 Joehrens K. V339 Johansson P. V786 Johrs C. P219 Jonat W. P599, V386 Jones D.T.W. V78 Jordan K. P549, P946, V96, V97, V101 Jost E. P194, P208, P563 Jost P.J. V675, V769 Jühling A. P862, P866 Jülich A. V795 Jung G. V354, V465, V1019 Jung J. V677 Jung M. P227, P939, V101 Jung W. P546 Junghanss C. P268, P269, P518, P548, P575, P579, P580, P585, P588, P589, P607, P609, P610, P848, P849, P964, V29, V50, V643, V478, V791, V793, V999 Jurczak W. P246 Just M. P604, V693 Jutz S. P272 K Kaemmerer D. Kaestner L. Kahl C. Kahl M. Kahlert L. Kähnert H. Kaiser F. Kaiser M. Kaiser N. Kaiser R. Kaiser S. Kaiser U.

Oncol Res Treat 2016;39(suppl 3):1–348

P294, P923 V476 P582, P876, V29, V792 P500 V672 V52 P581, P583, P940 P506 P312 P267 P602, P939, P943 V643

336

CONTENTS AUTHOR INDEX

Kalanovic D. P531, P880, P881, P882 Kalantari P. V995 Kalbacher H. P954 Kalias S. P613 Kalipciyan M. P896 Kalisch A. V95 Kalkreuth J. V46 Kallendrusch S. P312 Kalmanti L. P201 Kalmus J. P304 Kalupa M. V438, V439 Kambartel K. P568 Kaminsky B. V134 Kamolov B.S. P877 Kampa-Schittenhelm K. P186, P187, P504, P505, P506, P507 Kämpfe D. P215 Kandabarau S. P192 Kanji A. P248 Kantarjian H.M. V658 Kanz L. P186, P187, P192, P225, P263, P264, P271, P279, P285, P497, P505, P533, P546, P551, P552, P553, P560, P915, P968, V56, V57, V58, V88, V89, V145, V354, V388, V465, V683, V728, V1019, V1020 Kanzler S. V643 Kappeler C. P304 Karatas A. P295 Karawajew L. V634 Karch H. P198 Karcher A. V171 Kardošová M. V660, V831 Karn T. P604 Karpukhin A. P877, P874, P912 Kartal-Kaess M. P217 Karthaus M. P302, P303, P953, V44, V645 Kashkar H. V717 Kashofer K. V482 Kasper B. V321, V989 Kasper S. P304, P572, P614, V997 Kastritis E. V392 Katus H. V664 Katz J. V730 Katzerke C. P495, V660, V831, V834 Kauff F. V390 Kaufmann J. V646 Kechter A. V830 Keilholz U. P281, P565, P572, P593, V339 Keizman D. V1031 Keller A. V123 Keller L. P599 Keller P. V769 Keller U. P927, V688, V716, V718, V729 Kellermann L. P540 Kellner C. V659, V671 Kelly A. P941 Kemele M. P245, V720 Kenner L. V788 Kerkmann M. P948 Kern W. P191, P520, V142, V353, V637, V657 Kern Fürer C. S. 323 Kerstan H. P255 Kersten A. P563 Keßler C. P944

337

Kessler T. V989 Khaled N. V105, V728 Khan R. P508 Khan S.A. V672, V1018 Khandanpour C. V402 Khodamoradi Y. P849 Khoja O.T. P513 Kiani A. V675 Kiefer-Trendelenburg T. V48 Kiehl M. V685, P244, P252, V44 Kielstein J.T. V678 Kiesel M. P947 Kiewe P. P564 Killer M. V91 Killing B. P924 Kim H.-J. P257 Kim J. P888 Kim Y.-J. V720 Kimmel B. V75 Kimmich C. V661, V664 Kimmich M. P293 Kiote-Schmidt C. P906, V801 Kipkeeva F. P874 Kipps T.J. V34 Kirfel J. P884 Kirschbaum R. V771 Kirste T. V394 Kirsten N. V830 Kischel R. P280 Kischka T. V840 Kisro J. P208 Kissel S. V143, V733, V785 Kittel K. P604 Kiyota N. V998 Klaas O. V840 Klamroth R. P522 Klank D. P958 Klaproth H. V395 Klatt A.R. V717 Klauke K. V677 Klauschen F. P565 Klausmann M. P855, V675 Klausz K. V659 Kleber F. V634 Kleboth K. P307, P857, P951 Kleih M. P525 Klein C. P228, V686, V733 Klein F. P866 Klein I. P960 Klein S. P559 Klein S.A. P555 Kleine H.-D. P268 Kleiner H. P216 Klein-Hitpass L. V786 Kleinsimon I. S. 324 Kleist C. V466 Klenk J. P954 Klesse C. P516 Klimenkova O. P915 Klimiankou M. P192 Klingeberg C. V787, V833 Klinghammer K. P565 Klink A. P611, P909, V46, V441 Klinkhammer-Schalke M. V488 Klobuch S. P200 Kloss L. P190, V481 Klotz A. V1027

Oncol Res Treat 2016;39(suppl 3):1–348

Klug J. P943 Kluge A. P296 Kluger S. V104 Kluth S. P855, V685 Knauf W. P540, V390, V394, V846 Knaup-Gregori P. P238 Kneba M. P262, V393, V480, V684, V685 Knecht H. P262, V393, V684 Knecht R. V996 Kneis S. V173 Knittel G. V717 Knöbl P. V151 Knobloch U. P206 Knoll F. P270 Knoll N. V759 Knop S. P538, P545, P546, P606, V146, V666, V729, V808 Knorr P. V417 Knoth H. V180 Knübel G. P848, P849 Knyrim M. V348, V481, V783, V1012, V1013, S. 327 Kobbe G. P537, P559, V484 Koch B. P933, V763 Kochanek M. P197, V674 Koche R.P. V349 Köchel C. P606 Koeberle D. V342 Koehler M. V772 Koenecke C. V678 Koerfer J. P312 Kofla G. V44 Köhler A. S. 323 Köhler G. V987 Köhler K. P867 Köhler N. P549 Köhler P. V42 Kohlhase J. V759 Kohlhäufel M. P291 Köhn M. P501, P502 Kohnke J. P540 Köhnke T. P261, P280, V464 Köhrer S. V656 Koker M. P288 Kolanus W. V86 Kolarich D. P276 Kolatzki V. P519 Kolb M. V414, V417 Koleczko S. V134 Koller E. V358 Koller M. V488 Kollmeier J. P289 Konantz M. P884, V388, V405 Kondakci M. P537, V484 König J. V442 König P. S. 322 Königs C. P522 Konowski P. V46 Konschak R. V1000 Konstandin N.P. V356 Koopmann J. V684 Kopp H.-G. P225, P503, P533, P924, V56, V58, V989 Kopp J. P284 Köppler H. P307, P857, P951, P967 Körbel C. P245 Körbel O. P602

Author Index

CONTENTS AUTHOR INDEX

Korfel A. Korfsmeier K. Korn A. Körner S. Korotaeva A. Korovkina D. Körper S. Koschmieder A. Koschmieder S. Koser S. Kostenko A. Köster W. Koukakis R. Kowalewski D.J. Kozich Z. Krabisch P. Kragl B.

V82 P529 P264 V354, V465, V1019 P877, P912 V717 V625 P548 P511, V629, V771, S. 326 V809 P290, P860, V134, V988 P930 P302 V145, V683, V1020 P189 P604 P268, P269, P518, P548, P588, P589, V478, V665, V791, V999 Krahl R. V29, V178 Krajinovic K. P924 Kramer M. P194, V180, V181, V358 Krämer A. P217, V29, V67, V181, V458 Krämer C. V44 Krämer I. P944 Krammer-Steiner B. P521, P570, P579, P584, P594, P872, P961 Kräter M. P258 Krause D.S. V460, V640 Krause J. V1034 Krause S.W. P937, V155, V358, V725, V728, V749, V984 Krause T. V131 Krause-Kyora B. V1034 Krauss J. P572 Krauth M.-T. P240 Krebs J. P545 Krebs S. V32, V170, V347 Kreibich U. V29, V665 Kreil S. P219, P517, P555, V769 Krejci A. P262 Krekeler G. P531, P880, P881, P882 Kremer A. P895 Kremers S. V358 Krenosz K.J. P966 Kretschmer A. P276 Kretschmer D. P954 Kretzschmar A. V1022 Kreuter A. V342 Kreutmair S. V87, V787 Kreutz M. P188, P277, P895 Kreuzer K.-A. P197, V395, V658 Kreuzer M. V815 Kriegs M. V996 Kriegsmann M. P528, P532 Kriesen U. P579, P580, P585, P588, P589, P607, V478, V791, V793, V999 Krisper N. V482 Kristeleit R. V670 Kristen A. V661, V664 Kristensen B.W. P926 Kristiansen G. P884, V340 Krof-Sanchen C. P292 Krogel C. P917 Kröger N. P234, P559, V30, V45, V453, V485, V773, V781

Author Index

Krohn S. P964 Krohn-Grimberghe A. V146 Kron F. P290, P860, V988 Kröning H. V648 Kropf-Sanchen C. P287, P289 Kropp K. P285 Kropp P. V50 Kroschinsky F. P577, V756 Kruchen A. P919 Krug U. P197, P198 Krüger T. P258 Krüger W. P270, P514, P904, V440, V795 Kruggel L. V385 Krumm K. P530 Krupka C. P280, V464 Ksienzyk B. V356 Kuball J. V93 Kubin T. V358, V792 Kübler A. V465 Kubuschok B. V734 Kuendgen A. V805, V813 Kufer P. P280 Kühl A.A. P593 Kuhn M. P516, V756 Kuhn P. P292 Kuhn T. P602 Kühn M.W.M. V349 Kühn R.-B. P219 Kühn T. P604 Kühnel A. P230, P232, P233 Kukla K. P273 Kuklik N. V475 Kullmer J. V780 Kulozik A.E. V78 Kümmel S. V693 Kümpers P. P536 Kümpers S. P603 Kunde-Krüger J. P936 Kündgen L. P965 Kunert C. P909, P910 Kunitz A. P230, P232, P233, V731, V732, V989 Kunz C. P238, P944 Kunzmann V. V75 Kuon J. V792 Küpper M.K. P563 Küppers R. V786, V809 Kurbacher C. P602, P952, V693 Kurnik K. P522 Kürschner D. V358 Kurze S. V46 Kuß A.W. P249 Küster B. V729 Kuter D. P544 Kutsch N. V33 Kuzevanova A. P912 Kuznetsova I. P915 Kvalheim G. V464 Kvint R. P524 Kwee I. P527 Kyrtsonis M.-C. V391 L La Meir F. La Rosée P. Laack E.

P612 P204, P205, P206, P535, P959, V150, V155, V474 P287

Laban S. Lahner H. Lakis S. Lakner J. Lakner V. Lamm W. Lammer F. Lampe H. Lampe K. Lamprecht A.-K. Lancet J.E. Landschulze J. Lang A. Lang F. Lang G. Lang K. Lang S. Lange E. Lange F. Lange S. Lange T.

Langebrake C. Langer C. Langer W. Langerbeins P. Langguth P. Lanza F. Lapa C. Laryionava K. Laschke M. Lassmann S. Lathan B. Lauseker M. Lautermann J. Lazare S. Lazzaro A. Le D. Le Coutre P. Leblond V. Lechner D. Ledderose G. Lee P. Lee S. Leenders F. Legscha K.J. Lehmann H. Lehmberg K. Lehners N. Leibbrand B. Leiblein S. Leibold J. Leiser L. Leithäuser C. Leithäuser M. Leithold C. Leitner J. Leitzke S. Leleu X. Lemeer S. Lenard A. Lenerke C. Leng C.

Oncol Res Treat 2016;39(suppl 3):1–348

V996 P920, P921, P922 P291, V343 P518, P575, P609 P579 P240 P216 P588, P589 V101 V1024 P195 V31 P213 P558, V1015 P292 P516 V997 P211, V390, V685 V781 P268 P190, P207, P561, V155, V156, V418, V665, V726, V727, V976, V1012, S. 327 V45 P538, P546, V729 P241 V395 P274 P239 V688 V138, V760, V794, V979, V980, V981 P245 P913 P211, P220, P855 P201, V725, V728 P962 V677 P239 P888 P210, P266, V153, V154, V155, V770 V391 V442 P261 P293 V789, V841 P291 V667 V51 V474 P528, V43, V437, V674 V52 V467, V783 V465 P854 P579 P579, V643, V999 V96 P272 P267 V392 V729 P905 P905 P529

338

CONTENTS AUTHOR INDEX

Lenger D. P221, P966 Lengerke C. P496, P884, V388, V405, V842 Lengfeld T. V759 Lengfelder E. V358, V832 Lennerz V. P273 Lenz G. P198 Lenz V. V475, V477 Lerch J. P601 Lerchenmüller C. P220, P287 Leser U. V841 Lestin M. P579, P594, P961 Letsch A. P281, P578, V25, V779 Lettieri J.T. P941, P942 Leung A.W. P589 Leusen J.H.W. P276 Leuteritz K. V761 Leutz A. P889, V840 Levin C. P581 Li A. V135 Li J. V34, V391 Li L. V158 Liberati A.M. V770 Lichtenberger S. P944 Lichtenegger F.S. V464, P280 Lichter P. V78, V809 Lieberwirth B. P939, P943 Liebisch P. P546 Liebregts T. V758 Liedgens P. V717 Liedtke M. V658 Liesenjohann S. P537 Lietz C. P960 Liewald F. P292 Ligeti K. P562 Lin C.-S. P888 Lin T. P941, P942 Lindberg P. V488 Linde H. P855, P856 Lindl B. P280 Lindner A. P256 Lindner B. P934 Lindner C. V88 Lindner L. V989 Lindner S. P558, V1015 Link C. P516, P577 Link H. P944, P948, V358 Linné C. P577 Linnebacher M. P301 Linton K. V1007 Lipka D. V809 Lipp H.-P. V463 Lipp R. P309, P550, P861 Lipp S. V673 Lippert L.J. V723, V787 Lisenko K. P528, P532, P907, P908, V663 Liss B. V42 List C. P258 Liu Y. P494 Llombart A. P597 Lluch A. P597 Lodders S. P581 Löffler M.W. P551 Loges S. P604, V838, V985 Loghmani-Khouzani H. P915 Lohneis P. P862 Lohse S. P276 Loibl S. P604

339

Lonial S. Lopez J. Lopez R. Lopéz C. Löpprich M. Loquai C. Lordick F.

V730 V670 P597 P504, V717 P238 V792 P312, P873, V375, V434, V647, V792, V1022 Lorenz A. P952 Lorenz J. V134 Lorenzen S. P875 Lorenzen T. P576 Loscertales J. V141 Losem C. P211, P949, V648 Loskog A. V466 Loster A. S. 325 Loth F. P539 Lowinus T. P847 Lubberich R.K. S. 326 Lübbert M. V182, V350, V355, V746, V779 Luber B. V1022 Lubking A. P517 Lück A. P949, P964 Lücke S. P238 Luckhaupt H. P311 Ludwig H.P. V359 Ludwig W.-D. V371 Luebcke S. P273 Lueck C. V678 Lüers A. P296, V133 Luft T. V437, V1010 Lukas K. P281 Lundberg P. P496 Lupp A. P294, P923 Lutschkin J. P307, P857, P951 Lutz L. P913 Lutz M.P. V644 Lutz S. V671 Lux C. P872 Lyros O. V1024 M Ma S. V391 Maasberg M. P214 Maccari B. V93 Macdonald D. V391 Macherey S. V97 Machherndl-Spandl S. V358, V442 Mackensen A. P188, P229, P895, V144 Maddocks K. P246 Mader R.M. P896 Maertens J. P195 Mah N. V840 Mahler M. V141 Mahler S. V1025 Mahlmann S. P944 Mahon F.-X. V156, V675 Mährle T. V146, V985 Mai E.K. P238 Mai R. P923 Maier B. V646 Maier D. P224, V1022 Maier J. V727 Mairinger F.D. V339 Maixner F. V1034 Majidi F. P248

Oncol Res Treat 2016;39(suppl 3):1–348

Makałowski W. Makhro A. Maki R.G. Malehova O. Maleike H. Malenke E. Malvestiti S. Mamlouk K. Mancuso K. Mandic R. Mansmann U. Manso L. Manz M.G. Marc A. Marcello A.P. Marienfeld R. Mariotti E. Markhauser M. Märklin M. Markmann H.-U. Markova A.S. Marks R. Marmé F. Marquardt M. Marra A. Marreaud S. Marschner N.

V840 V476 V990 P874 P582 P503 V387 P863 P239 V1000 V356 P597 P265, P608 P561 V476 P292 P291, V343 P511 P225, V56, V57, V58 P870, P871 P877 P556 V693 P530 V987 V644 P295, P598, P869, V171, V174, V385, V693, V986, V1026 Martin H. P558, V1015 Martin P. P517 Martin R. P218, V404, V781, V782 Martin S. V358 Martinelli G. V658 Martinez J. P597 Martinkov V. P189 Martino B. V770 Martino S. P248 Marx G. V796 Marx J. P198 Maschke A. V769 Maschmeyer G. P546, P572, V29, V178, V474, V825 Maslyak Z. V34 Mässenhausen A.V. P884 Massuti B. V676 Mastris K. V1030 Mateos M.-V. V730 Mathas S. V789 Mathias E. V995 Matous J.V. V391 Mattern K. S. 325 Matthas S. P578 Matveev V.B. P877 Mauch C. V342 Mauer M.E. V644 Mau-Holzmann U. P504 Maurer C. P855, V685 Maurer S. V1019 Mayer J. V34, V141 Mayer K. P193, P543, P925, V674 Mayer R. P853 Mayer R.J. V645 Mayer-Steinacker R. V989 Mazieres J. V344 Mccarthy H. V34 McDermott D.F. P878

Author Index

CONTENTS AUTHOR INDEX

McGearey A. McGinnis C. McIntosh S. Meckel K. Meder L. Medinger M. Medyouf H. Meggendorfer M. Mehlis K.

V439 P905 P287 V674 P286, P288, V389 P496 V686 V104, V105, V637, V686 V138, V760, V794, V979, V980, V981 Mehnert A. P931, V792 Meincke M. P210, P211 Meinert F. V643, V989 Meisel R. V78 Meisner C. P546, V384 Meiss F. P913 Meiß F. V327 Meißner A.-M. V480 Meißner C. P876 Melemed S. P875 Melillo G. P572 Melisi D. P863, P865 Mellor-Heineke S. P192 Melmann A. P198 Mengwasser J. V438, V439 Menniger A. S. 323 Menon R. P291, V343 Mensen A. P282 Mentz K. P188 Merino J. V673 Merkel O. V788 Merkelbach-Bruse S. P568, P569, V134, V676, V843, V988 Merker N. V92 Merki R. V342 Mertens D. V809 Mertens T. V93 Mertlitz S. V438, V439 Merz F. P312 Merz M. P540 Messner C. V123 Mesters R.M. P198 Metz K. P614 Metzelder S.K. V156, V355 Metzeler K.H. V182, V183, V356 Metzgeroth G. P219, P605, P612, V104, V105, V127, V130, V401, V686, V768, V769 Metzner B. P546 Meulenbroek L.A.P.M. P276 Mey U. P534 Meyer D. V385 Meyer L. P576 Meyer L.H. V655, V656 Meyer R. P311 Meyer R.G. V355 Meyer S. P276 Meyer zum Büschenfelde C. P236 Mezger J. V131, V358 Mi G. V131 Miao H. P288 Michalczyk S. P867 Michel C. V47 Michels B. V93 Michels S. P290, P300, P568, P569, V134, V676, V843, V988 Middeke J.M. P194, P258, V180

Author Index

Middeke M. Midic D. V31 Mielke S. V119, V666 Miething C. P242, P508, P913, V787 Miethke T. V92 Mihajlović J. P899 Mikesch J.-H. P198, V987 Milanovic M. V841 Miller J.S. P918 Miller K. P887, V1031 Miller M. V693 Miltner E. P903 Minami H. V998 Minichsdorfer C. P240 Minnema M.C. V392 Mione M. V405 Mirbeth C. V468 Miriyala A. P304 Mirra N. V476 Mirza N. P264, P553 Misiak D. P501, P502 Miskey C. V1016 Mitschke J. P508 Mittlmeier T. P610 Moch H. V388 Mödder M. V172 Möginger U. P276 Möhle R. V771 Mohr B. P194, V780 Mohren M. V665 Moi S. P229 Molife L.R. V670 Möller M. V123 Möller P. P292, V717 Monecke A. P312, V467 Monjezi R. V1016 Monsef I. P587 Montesinos-Rongen M. V717 Montraveta A. P525 Moodley Y. V1034 Moosmann A. V464 Morales S. P597 Morche M. P255 Moreau P. V730 Moreno L. V673 Mori H. S. 323 Moriggl R. V788 Morishima T. V88, V89 Morrison R. V670 Morschhauser F. V1007 Morse M. P888 Mossner M. V401, V686 Motabi I. P209 Mottok A. P231 Motzer R.J. P878, P879 Mougiakakos D. P188, P229, V144 Muckel E. V52 Mueller M. P256, V788 Mueller N. V103 Mueller N.J. P613 Mueller T.A. V833 Muenchmeier N. P213 Muetzel E. P555 Mügge L.-O. P239, P538, P542, P611, P909, P910, V665 Mulaw M. V183, V830, V835 Müller A.M. P265, P613, P608, V831

Müller C. P509, P510 Müller D. P860, S. 325 Müller D.J. V57 Müller F. V42 Müller H. P253 Müller I. P919 Müller J. P291, P950, V173, V343, V345 Müller J.P. V94 Müller K. P258 Müller L. P299 Müller L.P. P559, P562 Müller M. P225, P533, V474 Müller M.C. P210, V156, V675, V728 Müller M.R. P503, V56, V57, V58 Müller N.J. P608 Müller R. P911 Müller S. P227, P228 Müller T. V340, V723 Müller T.A. V733 Müller V. P604, V31 Müller-Hagen S. V385 Müller-Tidow C. P197, P198, P494, P501, P502, P562, P946, V96, V834, V987, V1013 Müller-Werdan U. V101 Mumm F. P261, V762, V794, V979, V980, V981 Mundhenke C. P599, V386 Muranyi A. V830 Murua Escobar H. P548, P585, P588, P589, P848, P849, P964, V793 Mustjoki S. V157, V724 Musto P. P239 Mütherig A. P258 Muzaffarova T. P874 Mytilineos J. V1011 N Na I.-K. P282, V1017 Nachbaur D. V442 Nachtkamp K. V313, V484 Nadine K. P860 Naegele M. V801 Nagel G. P292 Nagy Z. P246 Nakano K. V998 Namasu C.Y. P495 Narimanov M. P874 Nasri M. V88 Nathan P. P878, P879 Naumann N. V104, V105, V768, V769 Naumann R. V358 Nazeer Batcha A.M. V356 Neagoie A.M. P292 Nebel A. V1034 Neef U. P962 Neemann N. V133 Nelde A. V145 Nelk I. P617 Nemitz J. P579 Nerreter T. P272 Netchaeva M. P296, V133 Neubauer A. P592, V91, V156, V355, V728 Neubauer E. P923 Neuber B. V673 Neuchel C. V1011 Neuendorff N.R. V357, V789

Oncol Res Treat 2016;39(suppl 3):1–348

340

CONTENTS AUTHOR INDEX

Neuer A. Neuhof A. Neukirchen J. Neumann F. Neumann K. Neumann M. Neumann S. Neumann T. Neumann U.P. Neumeister W. Neves M. Nickel N. Niederwieser D.

V999 P531 P537 P229, P245, V715, V734 P268 V32, V347, V401 V1022 P270, V440, V795 P305, P308 P571 V1007 V59 P190, P495, P559, P561, V29, V90, V106, V155, V178, V348, V467, V481, V660, V665, V683, V727, V783, V831, V834, V989, V1011, V1012, V1013, V1020 Niehr F. V1000 Niemeier B. P952 Niemöller C. V182 Nieschwitz D. P941 Nilsson S. P887, V1031 Nimmagadda S.C. V107 Nimmrich I. P289 Nimtz-Talaska A. P521 Noack N. V90 Nobile F. P239 Nogai A. P230, P232, P233, P564, V731, V732 Nogova L. P290, P300, P568, P569, V134, V676, V843, V988 Nold P. V91 Nolte F. P219, P555, V358, V401, V686, V779 Nolting J. P229 Nordlinger B. V644 Nörenberg D. P850 Nowak D. P216, V401, V686, V832 Nowak V. V401, V686, V832 Nowak-Harnau S. P560 Nowe E. V761 Nübling T. V354 Nuciforo P. P597 Nusch A. P211, P287, P598, V693 O Oakes C. Obländer J. O'Brien S. Ochsenreither S. Odenthal M. O'Dwyer M. Oechsle K. Oehlke O. Oellerich T. Oelschlägel U. Oergel T. Oertel R. Oettle H. Offner F. Oh W.K. Ohly A. Ohtsu A. Oing C. Oldenburg J. Oldenburg M. Olfe L.

341

V809 V401, V686, V832 V658 P281, P565, P593 V717 V34 V792, V796 P913 V719 P194, P258, P498 P904 V180 P862 V34 V1030 P617 V645 V651 P522 P932 P282

Oliverira M. P597 Ölsner M. P854 Oltmanns A. P567 Oostendorp R. V406, V688, V839 Opeker K. P939 Opferman J.T. V387 Opitz B. V29 Ordemann R. P258 Oriol A. V391 Orlowski K. P566 Ortega V. P597 Ortiz Tanchez J. V32, V347 Ortiz-Cuaran S. P568, V843 Ortmann M. V717 Ortmann O. V792 Ortner P. P948 Oscier D. P852 Ostermann H. P523, P546, V984 O'Sullivan J.M. P887, V1030, V1031 Ott G. P292, P525, P535 Ott P.A. P888 Otte K. P919, V687 Otte P. P939, P943 Otto G.P. P956 Otto S. P959, V764 Ouyang H. P875 Overkamp F. P305, P308, P310, P604, V693 Owen C. V34 Özöncel O. P578 P Pabst C. Pabst T. Pacaud L. Paccagnella M.L. Pachlopnik Schmid J. Pachmann K. Pachmann U. Paczulla A. Paganelli G. Pagel S. Pahernik S. Paiva B. Pajtler K. Pal M. Palandri F. Pallasch C.P. Palme I. Palumbo A. Pan K.-T. Pannenbeckers M. Panse J. Pantel K. Panzer I. Papasotiriou I. Pardo Jimeno J. Pare L. Pareigis S. Park K. Parmentier S. Paschen A. Pasparakis M. Passey C. Passweg J. Paul S. Pauli C.

Oncol Res Treat 2016;39(suppl 3):1–348

V1013 P608, V355 P920, P921, P922 V658 P608 P897 P897 P496 V1031 P273 P886 V673 V78 V59 V770 V59 V401, V686 V730, V770 V719 V469 P287, P563, P573, V44, V126, V474 P604, V838 P284 P902 V838 P597 P936 V344 P535, P547 P273, P275, V469 V717 V730 P496, V122, V747 P562 P494, P501, P502

Pauligk C. P864 Pavel M. P920, P921, P922 Pavel P. P907, P908 Pavenstädt H. P536 Paz-Ares L.G. P293, V135 Pecher A.-C. P533 Pechtel S. V484 Peer Zada A.A. P209, P513 Peest D. P546 Peeters M. P303 Pei W. V639 Peinert S. P283 Peipp M. V659, V671 Pelicci P.G. V834 Pelz H. P212 Pelzer U. P862, P865, P866 Penack O. V438, V439 Penrod J.R. V341 Pereira E. V676 Perez A. P500 Perez-Hernandez D. V439 Perglerova K. P191, V353 Perner S. P884, V340, V388 Pérol M. P293 Persigehl T. V717 Peschel C. P235, P546, P927, V406, V688, V718, V729, V839 Pester F. V31 Peter B. V668 Peter K. P895 Peter N. V29 Peters C. P913 Peters G. P198 Peters J. V477 Peters K. P570, P961 Peters M.V. P877 Petkova-Kirova P. V476 Petrie K. P500 Petti M.C. P239 Petzer A. P213 Peyn A. P955 Pezzi A. P239 Pezzutto A. P230, P232, P233, P284, P529, P564, P578, V483, V731, V732 Pfaff E. V78 Pfarr N. P600 Pfeifer D. V182, V327, V350 Pfeifer H. V480, V709 Pfeifer K. P266 Pfeilstöcker M. V316 Pfirrmann M. P201, P202, V155, V156, V675, V725, V728 Pfister S.M. V78 Pfrepper C. P559 Pfreundschuh M. P245, V715, V720, V728, V734, V1005 Pfütze K. P600 Pham M. P552 Phelps C. V141 Philipp U. P512, P867 Pigneux A. P195 Pillai R.N. P888 Pink D. P514, V440 Pinotti G. P239 Piribauer M. P251 Pittrow D. P956 Pizon M. P897

Author Index

CONTENTS AUTHOR INDEX

Plass C. Plattfaut C. Platzbecker U. Platzer J. Plauth M. Pleß M. Plettenberg A. Pletz M.W. Plimack E.R. Podar K. Poenisch W. Pogorzelski M. Pohlen M. Polliack A. Pompe T. Ponath E. Pönisch W. Popov S. Porkka K. Porpaczy E. Porteus M. Pöschel S. Posdzich P. Poser I. Postina P. Postrach D. Potenberg J. Pott C. Potthoff K. Poyrkov S. Prager G. Prange-Krex G. Prat A. Prehn C. Preisler M. Prenzel R. Pressler J. Preuss K.-D. Prevalsek D. Price T.J. Prinzhorn W. Pristupa A. Pritsch M. Procopio G. Proetel U. Pronina I.V. Proske A. Prosper F. Provencio M. Prüfer S. Prümmer O. Prutsch N. Przekopowitz M. Przybylski G. Puckert F.M. Pukhta I.A. Pukrop T. Pulewka K. Punke C. Puppe F. Puthenparambil J. Pylypenko H.

Author Index

V809 P898 V180, V181, V314, V358, V402, V686, V729, V745, V779, V780, V781, V784 P280 P617 V783 P576 P959 P878 V387 P540 V997 P197, P198, P536 V34 V90 V669, V790 P190, P538, P561, V29, V348, V481, V665, V783, V1012, V1013 P544 V157 P240 P497 P954 P267 P205 V92 V639 P254 P207, V393, V684, V720 P598, P949, V174 P877 P890 V675 P597 V716 P578 P296, V133 V401, V686 P245, V720, V734 P261 V644 P216 V141 P238 V1031 V728 P877 V785 V673 P246 V47 V675 V788 V786, V809 P249, P514 P232 P513 P595, V80 V762 V999 P545 V43 V141

Q Qorraj M. V144 Quach H. V34 Quast T. V86 Quecke T. P898 Queisser A. P884 Quidde J. P933, V763, V820, V985 Quietzsch D. P924 Quintanilla-Fend L. V835 R Raab M.S. Raake P. Raanani P. Rabe M. Rabitsch W. Rachow T. Rack B. Radloff J. Radsak M. Radtke M. Radujkovic A. Rammensee H.-G.

P235, P541 V664 V770 P301 P240, V151 V46, V674 P604 V985 P274, V47, V358 P941, P942 V437, V1010 P264, P553, V145, V683, V1020 Rani James A. V347 Ranson M. V670 Rao A. V56, V58 Rasche L. V666 Rassaf T. V403 Ratei R. V840 Rathert P. V459 Rattei T. V1034 Rau B. V1022 Rauh C. P892 Rauh J. P953 Rauner M. V402, V784 Rautenberg C. V484 Ravandi F. P195 Rawat V.P.S. V835 Rawluk J. P287 Ready N. V135 Rebmann U. P880, P881, P882 Reck M. P293, V131, V135, V341, V676 Regitz E. P245, V720 Rehklau S. P580 Rehli M. P229 Reich S. V388 Reichardt P. P924, V989 Reichart A. P864, V1023 Reichert D. P210, P283, P566, P950 Reichert J. V179 Reichle A. P200, P914, V358 Reifenrath K. V29 Reigl T. P262 Reim R. V771 Reimann M. V717 Reimer T. P594 Rein M. P873 Reinacher-Schick A. V982 Reinartz S. P592 Reinel E. P192 Reinel H. V644 Reinhardt C. P286 Reinhardt H. P939, P943, V717 Reinhardt M. P296 Reiser M. P868, P869, P956

Reiter A.

P517, V104, V105, V130, V739, V768, V769, V771 Reiter M. V634, V669, V790 Reiter T. P240 Rempel E. V1032 Ren M. V1028 Rendeiro A.F. P852 Rendenbach B. P214 Renner K. P188 Renner P. V95 Renner-Sattler K. P277 Rentsch A. V1025 Renz N. V182 Renziehausen L. P566 Reschke D. P938 Respondek-Dryba E. P584 Reuter S. P893 Reyderman L. V670 Reyher-Klein S. P254 Rezai M. P604, P952 Rhode C. P197 Riba J. V182 Richardson J. P954 Richardson P. V730 Richter A. P849 Richter C. P312 Richter D. P600, P931 Richter J. V675 Richter S. P916 Rick O. V701 Ridwelski K. P876 Riedel J. P585 Riedel K. P542 Riederer C. P938 Rieger C. P523, V674 Rieger K. P232, V762 Rieger M. P262, V61 Rieke D. P565, V339 Riemann J. P558, V1015 Riemekasten G. P898 Riemenschneider M.J. V117 Riera Knorrenschild J. V1022 Ries I. P554 Riese C. V172 Riesenbeck D. P946 Riesner K. V438, V439 Riess H. P862, P866 Rinaldetti S. P201, V728, V1032 Ringhoffer M. P965, V358 Rinke J. P203, V31, V179, V345, V346 Ripley A.V. V1030 Rister B. V733 Ritchie E.K. P195 Ritter E. P901 Ritter M. P530 Ritthaler M. V759 Rittig S.M. P285 Rittmeyer A. P287 Rittweger M. P256 Rivera F. P302, P303 Rizvi N.A. P287, V135 Robak T. P246, V34 Rocchi S. P239 Röcken C. P599, V386 Röder I. V641 Rodewald H.-R. V639 Rodriguez A. P500

Oncol Res Treat 2016;39(suppl 3):1–348

342

CONTENTS AUTHOR INDEX

Roedel C. Roeper J. Roessler J. Roessner A. Roggenhofer S. Rohde C. Röhl H. Rolfs F. Roll C. Röllig C.

V339 P296, V133 V639 P917 P591 P494, P501, P502 V686 V1024, V1027 P582, V643 P194, P235, V32, V180, V181, V358, V780 Römmler-Zehrer J. P864 Ronco J.P. V770 Roolf C. P848, P849 Rösch K. V642 Rosell R. V676 Rosenberg J. P888 Rosenwald A. P231, V63 Rösinger S. P896 Roskos M. V106 Rösler W. P559 Rosmolen J.C. P224 Roßberg A. P217 Rössig C. V79 Roßkopf S. V659, V671 Rössle S. V384 Rota P. V634 Roth A. V59 Roth M. V459 Röth A. P222, P223, V475, V477 Rothe M. P280, V464 Rothenberg-Thurley M. V356 Rothfelder K. P496 Rothmann F. V474 Rothschild S. V342, V676 Rotmann A.R. P586 Rotter N. V442 Rottorf M. P590 Rubanov O. P219, P306 Rubio I. P597 Ruch M. P939, P943 Rücker-Braun E. P516 Ruckser R. P213 Rudelius M. V729 Rudolph J. V86, V108 Rudolph L. V31 Rudolph R. P241 Rudolph S. P876 Rudorf A. V833 Rudzki J. P858 Rüegg C. P613 Ruf F. V839 Ruhparwar A. V664 Rule S. V141 Rummel M. V390 Rummler S. P892, P901 Rumpold G. V124 Ruppenthal S. P216 Ruppert A. V809 Rupprecht M. P287, P572 Rupprecht S. P210, P211 Russ J. V1017 Rüssel J. P572, V96, V643, V989 Rütjes A. S. 324 Rydzek J. P272

343

S Saad F. Sabranski M. Sack U. Sackmann A. Sadjadian P. Sadullah S. Saglio G. Sahlmann J. Sahm S. Salama A. Salat C. Salewsky B. Salih H.R.

P887, V1031 P576 V638 V1015 P519 V392 V158 P295 P305, P308, P310 V479 P598, P952 S. 326 P279, P285, P496, V145, V354, V355, V358, V683, V465, V1019, V1020 Salman M. V141 Salwender H. P236 Samareh B. P915, V88 Samek M. V45 Sammt A. V107 San Miguel J. V673 Sanda T. V788 Sändig I. P298 Sandner J. P578 Sandner R. P950, V986 Sänger J. P923, P294 Santucci Silva R. V141 Sasaki T. V998 Sasse B. P572 Sattler M. V387 Sauer A. P210, P211, P220, V154 Sauer M. V358, V780 Sauer S. V663 Sauerborn P. P257 Sauerland M.C. V356 Sauerwein W. P614 Saul D. V144 Saur S.J. P225, V56 Saußele S. P201, P207, V15, V153, V154, V155 ,V156, V675, V725, V726, V728 Sauteur L. P905 Savage P. P279 Savulsky C. V670 Sawall S. V838 Sawyer M.B. P941 Sayehli C. V780 Sayer H.G. P611, P909, P910, V29, V441 Schaab R. V781 Schaaf L. P525 Schaaf S. V356 Schadendorf D. P273 Schäd-Trcka S. P575 Schaefer-Eckart K. V75 Schaefers M. V643 Schäfer B. P944 Schäfer H.S. P247, P556 Schäfer S. P300, V59 Schäfer S.C. V717 Schäfer T. P884, P943, V388, V842 Schäfer V. P203, V179, V345, V346, V726 Schäfer-Eckart K. P590, V44, V329, V358 Schäffer M. P870, P871 Schafhausen P. V675, V779

Oncol Res Treat 2016;39(suppl 3):1–348

Schaich M.

P535, P547, P588, P589, P929 Schalk E. P957, V44, V674 Schaller G. V646 Schanz J. V779 Schanz U. P259, P265, P608, P613 Schaper F. P510 Schäpers C. V134 Schardt C. P210, P212 Schauwecker P. V93 Scheel A. P300, P568, V717, V843, V988 Scheffler M. P290, P300, P568, P569, V134, V676, V843, V988 Scheffold A. V656 Scheibenbogen C. P281, P282 Scheid C. P237, P267, V771 Scheidegger C. P283 Scheiner-Sparna R. P869, V1026 Scheithauer W. P890 Schellongowski P. V151 Schem C. P604 Schemionek M. P563, V620, V683, V1020 Schemmel L.-K. V666 Schemuth H. P222 Schenk T. P207, P500, V155, V157, V474 Schenke-Layland K. P954 Schepers H. V677 Schetelig J. P194, P282, P516, P559, V180, V181, V694, V756 Scheuer B. V61 Scheurlen M. P870, P871 Schewe D.M. V659 Schewe K. P576 Schichtl T. P602 Schieferdecker A. P234, V996 Schierle K. V1022 Schild H. P274, V47 Schildmann J. V374, V982 Schill C. V342 Schiller M. V43 Schilling A. P916 Schilling G. V763 Schilling J. P953 Schilling K. P611, V441, V444 Schindl M. P890 Schirmacher P. P600, V467 Schittenhelm M. P186, P187, P504, P505, P506, P507 Schlaak M. V342 Schlag P.M. V840 Schlag R. P511 Schlattmann P. V674 Schlee C. V32, V347 Schleef M. V1016 Schlegelberger B. V105 Schlenk R.F. P292, V358, V771, V779 Schleusener A. P578 Schlichting A. P306 Schliemann C. P198, V987 Schliesser G.C. V390 Schliffke S. V146, V985 Schlimok G. P197 Schlitt M. P262 Schliwa T. V665 Schlosser P. V350

Author Index

CONTENTS AUTHOR INDEX

Schlosser T. Schlößer H.A. Schmalbrock L.

P247 P893 P190, V348, V783, V1012, S. 327 Schmalenberg H. V435 Schmalz P. V676 Schmeer M. V1016 Schmeier-Jürchott A. V674 Schmelzle B. P292 Schmid B. V729 Schmid C. P197, P559 Schmid H. P279 Schmid K.W. V997 Schmid S. V442 Schmid T. V342 Schmidl C. P852 Schmidt A. P283, V842 Schmidt B. P219 Schmidt C. V973 Schmidt C.A. P249, P514, P969 Schmidt H. V101 Schmidt K. V437 Schmidt L.H. V987 Schmidt N. P582 Schmidt P. V644 Schmidt P.V. V484 Schmidt R. P903 Schmidt S. P213, P610 Schmidt T. P599, V386 Schmidt V. P611, V795 Schmidt-Hieber M. P230 Schmidtke-Schrezenmeier G. P292 Schmidts A. V87 Schmidt-Supprian M. V718 Schmidt-Wolf I. V170 Schmied B.J. V465 Schmiedgen M. P516 Schmiedl N. P231 Schmiegel W. V647 Schmier J.-W. V663 Schmier M. P965 Schmiester M. S. 326 Schmitt A. P907, V93, V466 Schmitt C.A. P850, P894, V717, V789, V841 Schmitt K. P573 Schmitt M. V93, V466 Schmitt S. V686 Schmitt T. P260, P554 Schmitt W. V982 Schmitz I. P847 Schmitz M. P258 Schmitz S. P220, P574 Schmohl J.U. P918 Schmoll H.-J. V643, V644, V989 Schmutz M. P612 Schnabl S. V669, V790 Schnappauf B. P604 Schneeweiss M. V668 Schneidawind C. P263, P271, P279, P497 Schneidawind D. P263, P271, P279, P497 Schneider A.S. V678 Schneider C.P. P287 Schneider D. P242, P508 Schneider E. S. 324 Schneider M. V106 Schneider R.K. V802

Author Index

Schneider S. V356 Schneider V. V1021 Schneidewind L. P270, V440 Schnell R. V171, V174 Schnelzer M. V815 Schnerch D. P891 Schnetzke U. P196, P851, V94 Schnitzler M. V171, V394, V986 Schnitzler P. V43, V93 Schnöder T. P509, P510, P847, V107 Schnorfeil F.M. V464 Schoenberg K. V108 Schoenberg S.O. V768 Schöffski P. V990 Scholl S. P196, P851, P910, P959, V29, V94, V355, V358 Scholten F. V648 Scholtysik R. V786 Scholz C.W. V64 Scholz M. V642 Scholz U. P522 Schön C. P965 Schönberger B. V769 Schönborn U. V107 Schönland S. V661, V664 Schormann C. V734 Schostak M. V1030 Schott D. P897 Schott M. P248 Schottelius M. V688 Schöttle J. P288 Schramm W. P522 Schrappe M. V659 Schraven B. P847 Schreck C. V839 Schreder M. P546, V666 Schreiber A. P969 Schrell S. P243 Schrenk K.G. P196, P959 Schrezenmeier H. V93, V475, V625, V1011, V1021 Schröck R. P601 Schröder M.P. P935, V32, V347 Schroeder T. P217, V355, V484, V813 Schroers B. P273 Schroers R. V474 Schubert A. P595, P945 Schubert J. V728 Schubert K. P190, V348, V481, V783, V1012, S. 327 Schubert M.-L. V466 Schuette W. P293 Schuh A. V34 Schuhmann L. V1012 Schuldt P. P286, V389 Schuler E. V779 Schuler M. P614, P916, V997 Schuler U. P258, P577, P916, V140 Schüler J. P891, V87 Schulte C. P215, P309, P550, P861, V134 Schulte E. P311 Schulz C. P261, V344 Schulz H. P949, P950 Schulz J. P190, P889, V348, V481, V783, V840, V1013 Schulz S. P294, P923, V764 Schulz X. V779

Schulze A. Schulze J. Schulze M. Schulze S. Schulze T.J. Schulze-Olden S. Schulze-Osthoff K. Schumacher H. Schumacher M. Schumacher U. Schumann C. Schumann L. Schumann U. Schumm M. Schuon A.-K. Schüssler L. Schuster H. Schütte W. Schütz C. Schütz E. Schwaab J.

V29 P932 P211, V440 V106, V178, V467 V401, V686 V134 V388 V50 P538 V103 P219, P293, V131 V783, V1013 V764 P551 V639 P603 V145, V1020 P962 V156 P848 V104, V105, V130, V768, V769 Schwaiger M. P927, V688 Schwammbach D. V987 Schwänen C. P292 Schwaner I. P309 Schwartz S. V347 Schwarz S. P864 Schwarzbich M.-A. P532 Schwarzer A. P550, P861, V665 Schwarzmeier J. P853 Schwind S. P190, P495, V348, V481, V660, V665, V783, V831, V1012, V1013, S. 327 Schwinger U. P210 Schwittay M. P305, P308, P310 Scriba D. P296, V133 Sebastian M. P295, V676 Seckinger A. V673, V807 Seeger J.M. V717 Seeling A. V180 Seevers R. P615 Seger M. V1031 Seggewiß-Bernhardt R. P287 Sehouli J. P593 Seibold M. P231 Seidel C. V1029 Seidel D. V42 Seifart U. P932, V700, V817 Seifarth W. P216 Seifert M. V786, V809 Seifert U. P847 Seitz J.-F. P304 Sekora A. P848, P849 Selivanova G. V655 Sell K. V478 Sellner L. P262, V466, V719 Sender A. V761 Senger J. P227 Sengül B. P593 Serke M. P287, P289, V134 Serva A. V809 Serve H. P194, P558, V32, V181, V358, V719, V1015 Severin K. P574 Sewtz C. V791 Seyfried F. V656

Oncol Res Treat 2016;39(suppl 3):1–348

344

CONTENTS AUTHOR INDEX

Sezer O. P546 Shah N.P. V158 Shah S. P907 Sharma P. P878, P888 Shehata M. V669, V790 Sheng J. V730 Shi Y. V438, V439 Shirneshan K. P218, V404, V779, V781 Shlyakhto V. V787 Shore N.D. V1030 Shoshani O. P234 Shoumariyeh K. P508, P512 Shumilov E. P218, V404 Shustik C. V391 Siddiqi T. V34 Siebert F. P251 Siebert R. V717, V786 Siebörger M. V468 Sieder C. P289 Siedler D. P221 Sieghart W. V669 Sigler C. P593 Sievers B. V404 Silin A. P189 Sill H. V482 Silling G. P563 Silva P. V32 Simeoni L. P509 Simon C. P264 Simon S.T. V27 Simon-Becker S. V404 Simon-Gabriel C.-P. V60 Simonis A. P608 Simpson D. V34 Singer K. P277, P895 Singh P. V391 Sinha A. V349 Sinn M. P862, P866 Siobal M. P288 Sippenauer T. V839 Sistermanns J. V792 Siveke J.T. P863, P865 Sklarz L. P848, P849, P964 Skoetz N. P237, P549, P587, V97, V170, V175 Skokowa J. P192, P915, V88, V89 Slabu I. P911 Slany A. P853 Slawska J. V716 Sleight B. V658 Sliwa T. P213 Slotta-Huspenina J. P927 Smetak M. V75 Smith U. P870, P871 Smola S. P270 Sobotzki C. V815 Sobrero A. P303 Socinski M.A. V131 Sockel K. V180 Soekler M. P187, P533 Soennichsen R. P312 Solem C.T. P865 Söling U. P598, V390 Sommer A. V342 Sommer C. P956 Sondermann P. P276 Song E. V349

345

Sonke G.S. Sopper S. Sørensen M.D. Sos M. Sosman J.A. Sotlar K. Spath C. Speith J. Sperr W.R. Spiekermann K.

V693 V157, V724 P926 P568, V843 P878 P520, V104, V105, V768 P886 V346 P213, V103, V151 P261, P280, V183, V356, V728, V832 Spiess B. V92 Spies-Weisshart B. V94 Spigel D.R. V135, V341 Spiliopoulou P. P888 Spira A.I. V344 Spittau G. V759 Spoerl M. P969 Spohn C. P856 Sprey C. P311 Spring L. P295 Springer G. P208 Springfeld C. P864 Spyroglou A. P248 Sreter L. P616 Srinivas S. P878 Staber P. V788 Stabla K. P592 Stachs O. P548 Stadler M. V678 Stahl M. P930 Stahlhut K. V792 Staib P. P530 Stalmach M. P903 Stamminger G. P226 Stangel M. P956 Starbatty B. P237, V170, V175 Stasik S. V181 Stassen M. P274 Stathis A. P527 Stauch M. P864 Staudinger T. V151 Staudt L.M. V717 Steckel N.K. P557 Stefan H. V156 Stefanzl G. V103, V668 Stegelmann F. P210, P517, V154, V158, V680, V728, V771 Steger G.G. P896 Steidler A. V1032 Steighardt J. V643, V989 Stein A. P933, V643, V644, V763, V985, V989 Stein H. P215 Steinacker J.M. V764 Steinbach-Büchert A.K. P305, P310 Steinberg T. P200 Steinberger P. P272, V788 Steinbrunn T. P231 Steiner M. P521 Steiner T. P880, P881, P882, P883 Steinger B. V488 Steinmetz T. P212, P220, P309, P574 Steins M. V135 Stelljes M. P197, P198, V658 Stemmler H.-J. P261 Stempelmann K. V1014

Oncol Res Treat 2016;39(suppl 3):1–348

Stengel R. Stengel S. Stenzel K.G. Stenzinger A. Stepien J. Steurer M. Stevanovic S. Stevens D.A. Stevens S. Stickel J.S. Stiefel O. Stilgenbauer S.

V106 P500 P305, P308, P310 P600, V1000 P601 P858, V319 V145, V683, V1020 V34 V792 V145, V683, V1020 V442 P243, V33, V395, V655, V656, V685, V741, V809 Stöbel-Richter Y. V761 Stock W. V658 Stocker G. V647 Stockhammer G. P858 Stoehr A. P576 Stöhr A. V401 Stölzel F. P194, P258, V180, V694 Stosch J.M. V182 Straka C. P546, V801 Strapatsas T. P537 Strassl I. V442 Sträter J. P292 Strathmann K. S. 326 Strauß-Rothenbücher A. V1023 Streckmann F. V51 Strefford J.C. P852 Strehblow M. V646 Strehl J.W. P214 Strein K. V673 Striefler J. P862, P866 Strobl C. P895 Stromberger C. V1000 Stropiep U. V133 Struckmann S. P848 Strunk D. V406 Strupp C. V313, V404, V697, V813 Strycker Orsini L. V341 Stuart R.K. P195 Stubbe F. P575, P607 Stübs P. P924 Stuck B. V997 Stuhler G. V666 Stuhlmann R. V403, V404 Stühmer T. P231 Stumpf T. V14 Stupp C. P574 Stuschke M. P614, V997 Stüwe S. V50 Styles L. V34 Subklewe M. P280, V356, V464 Südkamp N.P. P228 Süptitz J. V988 Suri D. V34 Sutter U. V771 Suttorp M. P915 Suzuki T. V998 Sweeting L. V670 Swords R.T. P500 Sy O. V730 Symeonidis A. V391 Sztankay M. V124 Szymaniak-Vits M. P939, P943 Szyska M. P282, V1017

Author Index

CONTENTS AUTHOR INDEX

T Tahara M. Tailor I.K. Takahashi S. Takats A. Tallo Parra M. Tam C. Tam C.S. Tameizi W. Tannir B. Tarantelli C. Targosz B.-S. Tashkandi S. Taube F. Tauchert F.K. Tausch E. Tavares R. Taylor F. Te Kronnie G. Tebbe S. Tebbutt N.C. Tebinka-Olbrich A. Tedeschi A. Teich M. Teichmann L.L. Teipel R. Teleanu V. Tenen D.G. Terwey T. Tesch H.

V998 P209 V998 P616 V781 V391 V34 V1023 V770 P527 V729 P209, P513 V181 V1023 P243, V395, V655 V770 P879, V341 V655 P211 V644 P945 V34, V391, V392 P306 P193 P258 P965 V831, V834 P266 P208, P210, P211, P212, P215, P220, P511, P602, P604, V174, V416, V417 Teschner D. V47, V672, V674 Tessen H.-W. P297, P306, P856, V986 Teutloff C. P537 Thaler J. P213, V648 Tharun L. P286 Thatcher N. V131 Thelen M. V342 Thellenberg Karlsson C. V1031 Thenhausen T. V146 Theobald M. P260, P273, P274, P275, P554, V47, V667, V672, V1018 Theurich S. P893, V342 Thieblemont C. V1007 Thiede C. P194, V32, V181, V358, V780 Thiel A. P277 Thiel E. P281 Thieltges F. P193 Thieme R. V1024, V1027 Thilo N. P955 Thölking G. P536 Thomalla J. P307, P857, P951, P967 Thomas M. P289, P295, V131, V676, V792 Thomas R. P288, P300 Thomas S. P200, V468 Thomsen A. P227, P228 Thomssen H. P955 Thornton N. V60 Thudium J. P197 Thurat M. P300, P569, V134, V676 Thurner L. P245, V720 Thuß-Patience P. P875 Tibor V. P927 Tichelli A. V122 Tiede A. P522, V627

Author Index

Tiemann M. Tienken M. Tietze-Bürger C. Tigges C. Timmer K. Timmermann B. Tinchon C. Ting S. Tinhofer I. Tischer J. Tischler H.-J. Toda K. Toenges R. Tombal B. Tomita Y. Tomka M. Tomlinson B. Tomska K. Torner J. Tournilhac O. Trajanoski Z. Trájer E. Trarbach T. Trautmann H. Trautmann K. Trebicka J. Trenschel R. Treon S. Trilling M. Trinks U. Tripp J. Tropashko I. Trotman J. Truckenmüller F.M. Trümper L. Trumpp A. Tsakiris D. Tsamadou C. Tschaika M. Tschechne B. Tucci M. Tunn P.-U. Turaev D. Turki A.T. Türkmen S. Turner S. Tykodi S.S. U Udonta F. Ueckeroth F. Uenalan M. Ufen M.-P. Uharek L. Uhl W. Uhlig J. Ullrich A. Ullrich J.G. Ullrich R. Ulshöfer T. Unseld M. Untch M. Unterhalt M. Unverzagt S. Uppenkamp M. Urlaub H.

P296, P309, V133 V792 P282 V342 P575, P607 V323 P213 V997 P565, V339, V1000 P261, P559 P540, V358 V998 P521, P570 V1030 P878 P251 V1030 V719 P580 V391 V157, V724 V764 P952, V385 V480 V1025 V86 P557 V391 V1014 P909 P601 P189 V391 V56, V58 V779 V686 P496 V1011 P888 P960 P887, V1031 V840 V1034 P557 V32 V788 P878 V838 V134, V988 P192 P299 P559 V982 P530, V665 V796 V770 P286, P288, V389 P210, P211, P602 P890 P604 V393, V973 P946, V96 P958 V719

V Vach W. V123 Valdix A. P306 Valent P. V103, V105, V668, V768 Valentini L. V763 Valerius T. P276, V659 Valladares-Ayerbes M. P302, P303 Vallera D.A. P918 Vallet S. P886, V387 Van Bebber F. V729 Van Cutsem E. P304, V644, V645 Van der Kuip H. P525 Van Lengen R. P944 Van Oorschot B. V792 Van Roye C. P307, P857, P951, P967 Van Tilburg C.M. V78 Vandendries E. V658 Vankann L. P573 Vannucchi A.M. V770 Vansteenkiste J. V344 Varga Z. V388 Vassiliou G.S. V349, V833 Vatter S. V468 Vegi N. V183, V835 Vehling-Kaiser U. P581, P583, P940, P963 Vehreschild J. V42 Vehreschild M. P300, V42, V44 Veratti P. P242, P508 Vercellati C. V476 Verga Falzacappa M.V. V834 Vergoulidou M. P250 Verhoef G. V1007 Verma U. P304 Vetter K. V771 Vettorazzi E. P604 Viardot A. P243 Vieth S. V659 Vij R. P544 Vilne B. V839 Vitolo U. V1007 Vlantis K. V717 Vlasic I. P286 Vogel A. P920, P921, P922, V643 Vogel W. P264, P551, P552, P553, P560, P884 Vogt J. V792 Vöhringer M. P525 Voi M. P920, P921, P922 Voigtländer M. V996 Vollmann J. V982 Volmer D. P229 Volz C. P286, P288 Von Amsberg G. V687 Von Bergwelt-Baildon M. P300, P893, V342 Von Bonin M. P258, V180 Von Bubnoff D. V327 Von Bubnoff N. P512, P867, P913, V327, V492 V771, V785 Von Deimling A. V78 Von Eggeling F. P911 Von Einem J. P572 Von Grundherr J. V763 Von Harsdorf S. V762 Von Hoff D.D. P863, P865 Von Kalle C. P600 Von Kries J.P. V789

Oncol Res Treat 2016;39(suppl 3):1–348

346

CONTENTS AUTHOR INDEX

Von Lilienfeld-Toal M.

P543, P892, P901, P909, V46, V421, V674 Von Minckwitz G. P604 Von Moos R. V648 Von Pawel J. P287, P289 Von Saint-George T. P929 Von Stackelberg A. V78 Von Stebut-Borschitz E. P274 Von Tresckow J. V395 Von Verschuer U. V986 Von Wussow P. P283 Vordermark D. V101 Vosberg S. V32, V347 Voskanyan A. P201 Voskova D. P221 Vossebein I. V997 Vu M.D. V673 Vucinic V. P190, P561, V29, V348, V481, V683, V783, V1012, V1013, V1020 Vuong L. P230, P266, P564 W Waanders M. P592 Waechter M. P230, P233, V731, V732 Wagner B. V464 Wagner E. V1011 Wagner E.M. P260, P554, P934, V672 Wagner K. P892 Wagner W. S. 326 Wahdat R. P220 Waidmann O. P864 Waizenegger J. V838 Waldau A. P203, V31, V179, V346 Waldschmidt J. P227, P228, V123, V801 Walenda T. S. 326 Walewska R. P852 Walker M. V90 Waller C.F. P210, V156, V675, V728 Wallstabe L. P272 Walsh N. V343 Waltenberger J. P197 Walter R. V719 Walz S. V1020 Wan Y. P865 Wang B. P288 Wang H. P884, V388, V842 Wang K. V658 Wang L. V93, V466 Wang Q. V809 Wang S.-Y. V106, V178 Wang T. V658 Wang Y. V156 Wang-Gillam A. P863, P865 Wanner D. P858 Wappl M. V646 Ward R. P195 Wardelmann E. P924 Wäsch R. P227, P228, P538, P556, P891, P906, V87, V123, V733, V801 Waterkamp D. V344 Waterstradt M. P904 Wattad M. V358 Waxman I.M. P878 Weber C. P256 Weber I. V1018 Weber M. P920, P921, P922 Weber T. P559, P562, V474, V674

347

Weddeling B. P913 Weersing E. V677 Weger R. P539 Wehler T. P287 Wehmeyer J. P511 Wehner R. P258 Wehrle A. V173 Wehrle J. P913 Wei A. P195 Weichert W. P600, V66, V339, V1000 Weickert M.-T. V406 Weide R. P214, P307, P857, P868, P949, P951, P967, V390 Weidmann-Hügle T. V100 Weidner H. P190, V402, V784, S. 327 Weidner-Zellmer L. P584 Weigert O. V970 Weigl R. P890 Weinelt D. P235 Weinert S. V107 Weis C.-A. V1032 Weise M. V756 Weisel K. P235, P263, P968, V662, V730, V1020 Weiß B. P958 Weiss H. P251 Weiß S. P570 Weisser B. P599, V386 Weissinger F. V1022 Weissmann R. P249 Welke C. P292 Wellhäusser U. P870, P871 Welslau M. P566, P883 Welte K. P192, P915, V88, V89 Weltermann A. P213, V442 Wendelin K. P590 Wendtner C.M. P855, V44, V395, V685 Wenthe J. V466 Wenzel F. P515 Wermke M. P258, V804 Werner J. P946 Werner M. P913 Wesseler C. P291 Wessendorf S. P292 Wester H.-J. P927, V688 Westermann J. P284, P850, V357, V635 Westhofen G. V403 Westner B. P234 Westphal S. P617 Wethmar K. P889, V840 Wetzel N. V171 Whetton A. V90 Wichelhaus A. P610 Wickenkamp A. P944 Wicki A. V335 Wicklein D. V103 Wider D. P227, P228, P891, V87 Widera L. P562 Widmann T. P591 Wiekhorst F. P911 Wierecky J. P938 Wieschermann U. P935 Wiesholzer M. P213 Wiesmüller L. V655 Wiesneth M. V93, V1021 Wiest G. P291 Wiestler B. P927

Oncol Res Treat 2016;39(suppl 3):1–348

Wiewrodt R. V987 Wiggermann P. V69 Wildenberger K. P190, V727 Wilhelm M. P590, P924, V75 Wilhelm S. P297 Wilk M. P537 Wilke H.-J. P875 Wilke J. V693 Will A. P587 Will U. P924 Willborn K. P296, V133 Wille K. P519 Willenbacher E. P539 Willenbacher W. P539, V392 Willschrei H.-P. P930 Wilop S. P563 Wimmer J. P229 Windemuth-Kieselbach C. V648 Winderlich M. P235, P246 Winiger I. P213 Winkelmann C. V665 Winkelmann N. P910, V179, V345 Winkler E.C. V138, V760, V794, V979, V980, V981 Winter K. P312 Wintner L.M. V124 Wirsching M. V123 Wirth M. P887, V1031 Wirth T. V671 Wirths S. P560, V56, V57, V58 Wirtz H. P298, P567 Wislocka L. P862, P866 Wisplinghoff H. V42 Witt O. V78 Witt R. V78 Witt S.H. V686 Witte K. P187 Wittekind C. P312, V1022 Wittel U. P867 Wittig J. V686 Wittke C. P268, P269, P518, P609, P964 Wittmann G. P523 Witzens-Harig M. P528, P532, P907, V432, V663 Wobus M. P498 Wöckel A. P587 Woelfel C. P273 Woelfel T. P273 Woerns M.-A. P864 Wohlfarth P. V151 Wohlleber E. V759 Wohlmann A. P205 Wöhrl S. P939 Woike M. P531, P880, P881, P882 Wolf D. P257, V86, V108, V110, V157, V358, V619, V724 Wolf H.-H. P546, V29 Wolf J. P290, P300, P568, P569, V134, V170, V676, V843, V988 Wolf S. P600 Wolf T. P869, V1026 Wölfel C. P275, V469 Wölfel M. P275 Wölfel T. P275, V469 Wolfensberger N. V769 Wolff D. P260, P559, P934, V762 Wolff T. P950

Author Index

CONTENTS AUTHOR INDEX

Wölfinger P. Wölfler A. Wolfrum P. Wolleschak D. Wolschke C. Wolz O.-O. Wörmann B. Worst B.C. Worst T. Wroblewski M. Wu L. Wu Z. Wuchter P. Wuerstlein R. Wulf-Goldenberg A. Wunderlich F.T. Wünsch A. Wurm A.A. Wurm R. Wuttke M. X Xanthopoulos C. Xuan F. Y Yamazaki T. Yang Y. Ychou M. Yepes D. Yomade L.O. Yoo H. Yoon S.

Author Index

P260 P213, V482 P939, P943 P509, P917, V107, V771, V772 V30, V45, V485, V773 V717 P549 V78 V1032 V838 V788 V93 P532, P907, P908, V93, V466, V663 P597 P230 V59, V717 V123 P495, V660, V831, V834, S. 327 P566 P943 V401, V686, V832 V693 V998 P863, P865 P304 V719 P542 V466 V995

Yu C. Yu Y. Yue Z. Z Zabelina T. Zacherle T. Zahabi A. Zahn M.-O.

V723 V789 V723

V30, V773 P291 V88, V89 P305, P308, P310, P864, V171,V174 Zaidi S. P513 Zaiss M. V385 Zalcberg J.R. V644 Zander T. P290, P860 Zanella A. V476 Zaniboni A. P304 Zaninoni A. V476 Zannetti B.A. P239 Zapatka M. V809 Zaplatina A. P286, P288 Zaun S. V648 Zebisch A. V482 Zeck G. V30, V773 Zehnder-Kiworr C. P944 Zehrfeld T. V665 Zeidler C. P192 Zelent A. P500 Zellmaier E. V356 Zellmer L. P579 Zenger M. P191 Zentner A. P945 Zenz T. P262, V143, V713, V719, V786, V809, V972 Zeremski V. V772 Zermann D.-H. V49, V53 Zhao F. P273

Zhao H. Zhou F. Zhou X. Ziegenhain C. Zielinski C. Zierhut M. Zieschang K. Zikic A. Zimmermann A. Zimmermann K. Zimmermann N. Zimmermann S. Zimmermann Y. Zink A. Zinke F. Zinngrebe J. Zintl P. Zinzani P.L. Zipori D. Zippelius A. Zirlik K. Zjablovskaja P. Zober A. Zoellner A.-K. Zojer N. Zschäbitz S. Zu Eulenburg C. Zubairi I. Zuber J. Zucca E. Zügel M. Zur Hausen G. Zwerger M. Zwick A. Zyczynski T.

Oncol Res Treat 2016;39(suppl 3):1–348

P878 P494, P501, P502 P528 V406, V839 P890, V669, V790 P264 P191 P915 P293 P270, V841 S. 322 V182 P524, P526 V1034 P958 V656 P527 P246, V1007 P234 V342 V60, V143 V831 P538, V123, V801 P261 V29 P886 P604 V670 V459 P527 V764 P864 V468 V406, V733, V785 P544

348