Magnetotheranostics by Breadfruit

Magnetotheranostics From superparamagnetic nanoparticles to tools for the detection and treatment of cancer M. Mionic1, D. Bonvin1, H. Hofmann1, H. Richter2, B. von Rechenberg2, S. Barbieri3, H. Thöny3, J. Bastiaansen4, M. Stuber4, S. Ehrenberger5, O. Jordan5, G. Borchard5, M. Capstick6, E. Neufeld6, N. Kuster6 1EPFL, 2University of Zurich, 3Inselspital Bern, 4CHUV, 5University of Geneva, 6It’IS

Débora Bonvin, Institute of Materials, EPFL Nano-Tera Annual Meeting, 04th May 2015, Bern

Prostate cancer stages Primary tumors

Early metastases (lymph nodes)

Late metastases (bones, lungs, â&#x20AC;Ś)

PSA* levels (ng/ml)

< 20

> 20 or Any

Any

5-year survival rate

90-100%

85-100%

40-80%

MRI* (>8mm)

PET/CT***, MRI**

50% metastases are missed

Easy detection BUT often too late

Detection method

*Prostate specific antigen **Magnetic resonance imaging ***Positron emission tomography/computed tomography

Goetz, 2008

MagnetoTheranostics • Diagnostics of prostate cancer metastases in lymph nodes

(< 8 mm) by MRI with superparamagnetic nanoparticles Benign lymph node

Metastatic lymph node

Froehlich et al, 2012

 Difficult to distinguish healthy/metastatic part  Dark grey scale  Lack of specificity (< 100% metastatic lymph nodes)

MagnetoTheranostics â&#x20AC;˘ Diagnostics of prostate cancer metastases in lymph nodes

(< 8 mm) by MRI with superparamagnetic nanoparticles Benign lymph node

Metastatic lymph node

Froehlich et al, 2012

â&#x20AC;˘ Treatment of prostate tumors by hyperthermia o Metastases in lymph nodes o Primary tumors Andrade et al, 2011 Jordan et al, 2011

Engineering ; ITIS, ANTIA

In-vitro, Toxicity, imaging EPFL, ITIS, UNI GE, CHUV

Task 1 Existing and New Particle composition

Development Temp. Simulation tool Improvement of mag Generator

Task 3

Task 4

Nanocomposite formulation

Functionalisation of Particle with Antibodies

In vitro tests Heating capacity

In vitro tests Specific adsorption Metastasis

In vivo tests tumor treatment

st e T

Task 2

Characterisation Tox screening

icty x o T

Hyperthermia

Medical application CABMM, Inselspital

In vivo tests Theragnosis

In vivo tests metastasis detection

Molecular Imaging (MRI)

Physics, chemistry Material science; EPFL, UNI GE, CHUV

EPFL

Task 1: Development of nanoparticles

Properties of new nanoparticles that have to be fulfilled o Physico-chemical requirements o Biomedical requirements

Selected materials: o Development based on iron oxide (i.e. Îł-Fe2O3, SPIONs) o Exploration of new materials not yet used for medical applications

(e.g. doped transition metal oxides)

EPFL

Task 1: Synthesis of γ-Fe2O3 Magnetic core

- Suitable for MRI - High heating for hyperthermia

Specific loss power (SLP): W / gFe2O3

Brownian

Relaxation time τ

Néel

f and H are limited (50<f<12000kHz; 0<H<15kA/m; f*H < 485 kHz*kA/m)

η fluid viscosity, K particles anisotropy constant, Ho amplitude and f frequency of alternating magnetic field, μo magnetic permeability and Ms particles saturation magnetization

NANOPARTICLE SIZE!!!

EPFL

Task 1: Synthesis of Îł-Fe2O3 Synthesis fulfills the requirements for medical application o Aqueous o No organic solvents o No solvent exchange o No coping agents

o Simple o Reproducible, repeatable (100 repetitions) o Scalable

EPFL

Primary size (nm)

8.0 ± 1.9

14.7 ± 5

15.6 ± 4.7

19.0 ± 5.7

17.4 ± 4.7

21.5 ± 6.3

Crystallite size (nm)

8.7

13.2

13.7

17.8

18.2

Hydrodynamic size (nm)

17.7 ± 5.8

26.9 ± 8.5

29.5 ± 8.5

25.8 ± 7.8

35.1 ± 10.6

30.2 ± 9.1

Zeta potential at pH 4 (mV)

55.6 ± 0.4

47.4 ± 2.2

47.9 ± 2.3

46.3 ± 1.4

49.3 ± 2.4

48.2 ± 0.6

EPFL

Task 1: Magnetic characterization Magnetic susceptibility

Magnetization

EPFL

Task 1: SLP for hyperthermia 100kHz, 20mT (16kA/m)

ΔT (°C)

1 2 3 4 5 6

Time(s) Sample

Diameter (nm)

SLP W/gFe2O3

8.0

5,4

14.7

85,4

15.6

121

19.0

127,1

17.4

115,5

21.5

110,8

5mT 264kHz

30mT 96.5kHz

30mT 229kHz

30mT 248kHz

30mT 264kHz

30mT 314kHz

30mT 352kHz

30mT 440kHz

30mT 571kHz

Itâ&#x20AC;&#x2122;IS

Task 2: Development of generator

Magnetic field generator applicable to humans o Challenges and requirements o Coilsâ&#x20AC;&#x2122; size suitable for humans (bore diameter 400mm) o f = 300kHz: sufficient hyperthermia, but with clinically applicable conditions (f*H

< 485 kHz*kA/m)

Itâ&#x20AC;&#x2122;IS

Task 2: Development of generator

Magnetic field generator applicable to humans o Challenges and requirements o Low voltage/current + homogeneous field strength to avoid hot spots

sensitivity 84uT/A)

(field

Itâ&#x20AC;&#x2122;IS

Task 2: Development of generator

Temperature simulation software o Heating prediction (tumor, unwanted hot spots) -> safety and efficacy o New high-performance computing enabled thermal solver o Support of inhomogeneous heat source and tissue properties o Advanced perfusion models, generation of patient-specific vasculature

o Image-based heat source distributions o Information extraction from medical image data o Relationship between SPION density & field strengths and heating

University of Geneva

Task 3: Tumor treatment by hyperthermia

Implant for local treatment of primary prostate cancer BLADDER

SPIONs Radio-opaque polymer DMSO

solid implant TUMOR liquid formulation PROSTATE

Solidifaction upon contact with aqueous solution

Liquid formulation

Implant

Scanning electron microscopy of precipitated implant

University of Geneva

Task 3: Tumor treatment by hyperthermia Injectability (18wt% polymer)

Magnetically induced heat release

BLADDER

solid implant

100kHz, 20mT

45 1 2 3 4 5 6

SPIONs

liquid formulation PROSTATE

ΔT (°C)

TUMOR

Cell viability (WST-1 test) PC3 Fibroblast

7nm-SPIONs in composite (20wt%)

Time(min)

300kHz 12 mT 9 mT 6 mT 3 mT

CHUV, University of Zurich, EPFL, University of Geneva

Task 4: Detection of LN metastases by MRI

Functionalization of nanoparticles with: o Coating o Targeting molecule towards tumor as specific as possible

MRI imaging protocol o Development of new sequences o Improvement of SPIONs localization

CHUV, University of Zurich, EPFL, University of Geneva

Task 4: Functionalization (coating) Coating processing for medical application o Simple, scalable, aqueous medium, no solvent exchange

11 coating molecules o Biocompatible, as small as possible o Groups known to have high affinity to SPIONs o

Phosphate groups

o Carboxylic groups (e.g. folic acid)

o ‘’Available’’ functional groups for coupling with targeting molecules

Coating characterization o FTIR, UV-Vis, XPS, Zeta potential and size measurements (DLS),

High resolution TEM, Toxicity in vitro tests, TEM up-take study

CHUV, University of Zurich, EPFL, University of Geneva

Task 4: Functionalization (antibody) Requirements for biomedical applications o No use of organic solvents (only aqueous medium) o No use of catalysts o Targeting molecule has to stay active after coupling

Proof of principle of coupling o Coupling of SPIONs with IgG targeting Echinococcus o FTIR, ELISA SPIONs Coated-SPIONs EmG11-SPIONs Antibody-SPIONs

CHUV, University of Zurich, EPFL, University of Geneva

Active targeting of LN metastases of prostate cancer o Target: prostate specific membrane antigen (PSMA) o Targeting molecule: o

Humanized antibody

Small molecule

Aptamer (ssRNA)

SPION

CHUV, University of Zurich, EPFL, University of Geneva

Task 4: Novel positive contrast MRI Detection of SPIONs

IRON

o Improved contrast o Improved SPION localization o Removal of false positives

Mapping of SPIONsâ&#x20AC;&#x2122; induced dipolar field o Quantification of SPION concentration (heating)

IRON + UTE

Conclusion o SPIONs for theranostics  Maximization of heating for hyperthermia  Aqueous synthesis without any toxic solvents

o Prototype of hyperthermia generator  Size, frequency, current/voltage applicable to humans

o Suitable and injectable composite  Biocompatible, promising heating

o Successful functionalization  11 coatings in aqueous medium  Proof of principle of coupling with antibody

Thank you for your attention !!