Research Outreach Issue 102

The outreach quarterly connecting science with society ISSN 2517-7028 ISSUE 102

FEATURING RESEARCH FROM:

University of North Carolina, University of Texas at El Paso, Canadian Society for Molecular Biosciences (CSMB), The Archbold Biological Station, North Carolina State University, McGill University, Vanderbilt University School of Medicine, Canadian Blood Services, UniversitĂŠ de Sherbrooke, University of Chicago, Roswell Park Cancer Institute, University of British Columbia, University of Delaware, International Agency for the Prevention of Blindness (IAPB), The UCLA Henry Samueli School of Engineering and Applied Sciences, University Hospital Research Features 3 Carl Gustav Carus, University of California, Ludwig-Maximilians-Universitat, Brown University, Health Research Alliance (HRA)

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We abide by the Creative Commons (CC) license terms to ensure widespread, open-access dissemination of all the work featured across our various platforms.

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RESEARCH OUTREACH ISSUE 102

WELCOME

The public outreach magazine for the research community ISSUE 102

TO ISSUE 102

FEATURING RESEARCH FROM:

There are many overlaps between the fields of Health & Medicine and Biology. In this issue of Research Outreach, we feature researchers from both disciplines and hear from thought leaders across the subjects.

Dr Maryrose Franko, Executive Director of the Health Research Alliance explains why popularising research is so important and why the organisation puts people at the heart of its policies. Dr Philip Hieter, President of the Canadian Society for Molecular Biosciences (CSMB), emphasises the importance of advocacy and education for securing funding and support.

Canadian Blood Services’ CEO Dr Graham Sher explains the story behind the non-profit charitable organisation’s mission to effectively manage the bloody supply for Canadians. The International Agency for the Prevention of Blindness (IAPB) promotes eye health through advocacy, knowledge and partnerships. CEO Peter Ackland discusses their mission and the importance of World Sight Day. In addition to our Thought Leaders, you will find this issue packed full of fascinating research from Health & Medicine and Biology. Join our global readership and dive straight in to find out more!

University of North Carolina, University of Texas at El Paso, Canadian Society for Molecular Biosciences (CSMB), The Archbold Biological Station, North Carolina State University, McGill University, Vanderbilt University School of Medicine, Canadian Blood Services, Université de Sherbrooke, University of Chicago, Roswell Park Cancer Institute, University of British Columbia, University of Delaware, International Agency for the Prevention of Blindness (IAPB), The UCLA Henry Samueli School of Engineering and Applied Sciences, University Hospital Research Features 3 Carl Gustav Carus, University of California, Ludwig-Maximilians-Universitat, Brown University, Health Research Alliance (HRA)

THIS ISSUE Published by: Research Publishing International Ltd Publisher: Simon Jones Editorial Director: Emma Feloy emma@researchoutreach.org Operations Director: Alastair Cook audience@researchoutreach.org Editor: Hannah Fraser hannah@researchoutreach.org Designer: Craig Turl Project Managers: Kate Cooper (Senior) kate@researchoutreach.org Tobias Jones tobias@researchoutreach.org Ben Phillips ben@researchoutreach.org James Harwood james@researchoutreach.org Contributors: Alex Davey, Siobhan Fairgreaves, Ingrid Fadelli, Anna Jones, Karen O’Hanlon Cohrt, Kate Porter, Jasmin Skelly, Paul Smith, Victoria Stanley Tsui. /ResearchOutreach /ResOutreach

Editor Please feel free to comment or join the debate. Follow us on twitter @ResOutreach or find us on Facebook https://www.facebook.com/ ResearchOutreach/

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BUILDING INTERDISCIPLINARY BRIDGES TO STUDY INTERCELLULAR BRIDGES Dr Amy Maddox Uncovering the properties of intercellular bridges within multinucleated cells.

NOVEL 3D MICROSCOPE PROVIDES UNPRECEDENTED MOVING IMAGES OF BIOLOGICAL PROCESSES Dr Chunqiang Li A novel 3D optical microscope that uses a spectrally shaped pulse laser.

CSMB: ENSURING A STRONG FUTURE FOR SCIENCE ND SCIENTISTS Dr Philip Hieter The Canadian Society for Molecular Biosciences supports fundamental research.

FLORIDA’S ARCHBOLD BIOLOGICAL STATION GIVES ONLINE ACCESS TO UNUSUAL NATURAL HISTORY COLLECTION Dr Hilary Swain Making data and images of thousands of biological specimens available online.

COMMUNICATING THE LANGUAGE OF PLANTS THROUGH INOSITOL PHOSPHATES Dr Imara Perera Investigating signalling molecules to understand their role in plant communication.

CONTROLLING MAGNESIUM FLUX: A CENTRAL ROLE FOR THE PRL-CNNM COMPLEX Professor Michel L. Tremblay Investigating the role of a newly discovered pathway in controlling magnesium flux.

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BLOOD, SKIN AND BONE: THE COMPLEX CONTROL OF BLOOD PRESSURE Professor Raymond Harris and Professor Ming-Zhi Zhang Uncovering a novel role for bone marrow-derived immune cells in salt-sensitive high blood pressure.

CANADIAN BLOOD SERVICES: THE ROAD TO REGAINING PUBLIC TRUST Dr Graham Sher Providing a safe, secure, cost effective and accessible blood supply for Canadians.

DISORDERED FAT STORAGE ENHANCES DEVELOPMENT OF TYPE 2 DIABETES Professor André Carpentier Exploring how obesity increases the risk of type 2 diabetes

We identified a number of ‘neurobiological bricks’ that are central for cognitive control mechanisms.

PROFESSOR CHRISTIAN BESTE Page 68

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THE GENOMICS OF CANCER Dr Lixing Yang Uncovering structural DNA rearrangements.

REPROGRAMMING THE IMMUNE SYSTEM FOR PERSONALISED IMMUNOTHERAPY AGAINST CANCER Dr Richard Koya Using patients’ own immune systems to target and kill cancer cells.

INHIBITING CANCER STEM CELL SURVIVAL IN THE HOSTILE TUMOUR ENVIRONMENT Dr Shoukat Dedhar The possibility of using a cancer cell’s own physiology as a weapon against it.

THE EPIGENETIC EFFECTS OF ADVERSE EARLY-LIFE EXPERIENCES Dr Tania Roth Exploring the impact of the environment, particularly stress factors, on individual genes.

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IAPB: ENVISIONING THE FUTURE OF UNIVERSAL EYE CARE Peter Ackland Leading international efforts in blindness prevention activities and eye health.

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A NEW TERAHERTZ MEDICAL IMAGING TOOL COULD PROVIDE EARLY DETECTION OF CORNEAL DISEASE Dr Warren Grundfest and Dr Zachary Taylor A promising method to accurately detect and study cornea-related diseases.

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UNVEILING THE NEUROBIOLOGICAL PROCESSES BEHIND COGNITIVE CONTROL Prof Christian Beste Understanding the neural

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underpinnings of human goaldirected behaviour.

Understanding sleep disordered breathing (SDB) in pregnancy.

COMMUNITY COLLABORATIONS TARGETING BREAST CANCER Dr Marion Kavanaugh-Lynch Coaching for community and academic partnership teams conducting participatory research.

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HRA: MAKING SCIENCE OPEN, MANAGED AND WELL-FUNDED Dr Maryrose Franko The number one place for non-profit organisations seeking to enhance return on their investment in biomedical research.

DUAL ORIGINS OF TISSUE MACROPHAGES Professor Christian Schulz Demonstrating that a large proportion of the macrophage population is derived during embryonic development.

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COMMUNICATION Social media and the rise of video How video use is increasingly important on social media.

THE DANGERS OF SLEEP DISORDERED BREATHING IN PREGNANCY Professor Ghada Bourjeily

RESEARCH AREAS

Biology

Health & Medicine

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Biology ︹ Dr Amy Maddox

Building interdisciplinary bridges to study intercellular bridges Dr Amy Shaub Maddox from the University of North Carolina is leading a diverse team of researchers to uncover the properties of intercellular bridges within multinucleated cells. Known as syncytia, cells with interconnected nuclei can be found in model organisms such as Drosophila and Caenorhabditis elegans as well as more complex organisms, including humans. Dr Maddox has also placed significant emphasis on outreach to a variety of audiences throughout the course of this research. The imaginative techniques the team is using are sure to inspire successful collaborations within the university community and beyond.

T

he building block of all life is the cell. The architecture of a generic animal cell is well understood: a single nucleus resides in the cytoplasm, surrounded by the cell membrane.

However, not all cells are generic. Both simple and complex animals in fact have some specialised cells with more than one nucleus. A cell with multiple interconnected nuclei is known as a syncytium. One place in the body where syncytia are found is the tissues that generate sex cells (sperm and eggs). In

The syncytial germline of the nematode C. elegans, with DNA (blue) and the syncytial lining (magenta) labelled.

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these syncytia, nuclei are connected to a common cytoplasm via intercellular bridges. The work of Dr Maddox and her team of researchers at the University of North Carolina focuses on the composition, dynamics and regulation of these bridges.

SYNCYTIA: WHAT? WHERE? AND WHY? To understand more about the work of Dr Maddox and her colleagues, it is necessary to understand more about

Amy (left) in the microscope room with members of her lab.

the syncytium. Though syncytia are a little different from the commonly taught norm, they are present throughout the animal kingdom. From the fruit fly, Drosophila, to humans, many species feature syncytial cells in their germline. The germline of a multicellular organism is the population of its bodily cells which allows it to pass on genetic material via sexual reproduction.

The structure of syncytia, including the properties of the intercellular bridges, has important consequences for fertility. Intercellular bridges allow for communication and coordination among the nuclei that reside there together, incompletely partitioned like horses in barn stalls.

For example, in various types of syncytia, cytoplasm must flow out of some stalls and into others to enlarge them. When the oocytes are fully enlarged, the bridges collapse, thus achieving cellularisation.

If the bridges collapse prematurely, the oocytes do not enlarge properly, and are not viable. Thus, these intercellular bridges must be stable. However, Dr Maddox’s lab recently found that in the gonad of the model organism Caenorhabditis elegans, intercellular bridges are surprisingly dynamic

throughout the germline, enlarging and contracting multiple times over their lifetime.

How syncytia form and are maintained remains poorly understood, however, research by Dr Maddox and others has begun to unravel some of the mysteries. For example, some of her earlier work identified a protein called anillin-2, which promotes the integrity of syncytia and endows the intercellular bridges with elasticity and stability.

Dr Maddox’s lab has now discovered another pair of proteins that regulate the stability of intercellular bridges.

Interestingly, one is implicated in a human disease of brain vasculature, so

characteristics of this worm make it a powerful organism for syncytial studies; not least the well understood architecture of its simple body plan and invariant lineage of its cells. Similarly to other model organisms such as the fruit fly, genomic modifications of C. elegans can be conducted with relative ease.

their discoveries of its mode of action may help our understanding of that condition.

By manipulating the genome of C. elegans to introduce fluorescent protein tags, the team is using quantitative microscopy to gain unprecedented insight into the make-up of intercellular bridges. Quantitative microscopy allows for automated analysis of two- or three-dimensional images, meaning vast amounts of data can be analysed much more quickly than via manual measurement.

Quantitative analysis is powerfully combined with high-resolution light microscopy techniques housed in the Maddox lab and within her department’s shared equipment facility.

STUDYING SYNCYTIA The model organism for this study, Caenorhabditis elegans, is a tiny nematode worm with a history of providing answers to some of the big questions in modern science. In fact, work with C. elegans has garnered three different Nobel Prizes. Many

COLLABORATION ACROSS THE CURRICULUM The team of researchers led by Dr Maddox share diverse geographical and educational backgrounds and the scope of the project allows for the training and mentoring of individuals at several stages of their education,

The structure of syncytia, including the properties of the intercellular bridges, has important consequences for fertility

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from undergraduate students to postdoctoral fellows. Amongst the lab members involved in the “hands on” research are Daniel B. Cortes, Kathryn Rehain-Bell and Michael Werner. Dr Maddox benefitted from the fact that the other researchers involved with this study are dedicated to and experienced with educational outreach.

These activities ranged from interactive sessions with elementary school children to lab tours for prospective undergraduates.

Using the project to engage multiple audiences was very important to Dr Maddox. Her outreach activities sought out different groups within the University of North Carolina as well as individuals and groups in the local community.

One of the groups Dr Maddox was keen to involve with this project was the local education sector. She approached this aim in two ways. Firstly, each summer a teacher is invited to participate with the lab work for a 3-5 week internship.

Dr Maddox hopes the experience will inspire these teachers to take their experience back to the classroom and relay their new knowledge and enthusiasm to their pupils. Secondly, Dr Maddox visits an elementary school to teach children about embryonic development. This classroom based project was developed according to established teaching goals and with the school’s science specialist.

Dr Maddox also wanted to make the most of the already-collegial campus environment at the University of North Carolina by creating innovative opportunities for engagement. In order to improve interdisciplinary collaborations between the fields of biology and mathematics, Dr Maddox developed a module that allowed students from these courses to work alongside each other. In the module, the students experience novel methods of quantitative analysis and computer modelling using data they collect in the lab, studying intercellular bridges.

One of the most original pieces of engagement proposed by Dr Maddox was interdisciplinary speed-dating.

During these sessions, biologists briefly present projects to mathematicians,

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C. elegans hermaphrodite

Oogenic germline

Intercellular bridges

Intercellular bridge regulation

Ste20 Kinase

GCK-1

CCM-3

non-muscle myosin II

anillin

Schematics of progressively magnified views of intercellular bridges (blue circles) in the C. elegans syncytial germline.

Our outreach activities will strengthen scientific research potential, not only during the funded period but for generations to come physicists, computer scientists and statisticians interested in collaborations.

Several successful partnerships have emerged from these events, and still more participants benefit just from the time spent building community.

Both the scientific and public engagement elements of this project provide numerous opportunities for

future research. Dr Maddox is confident that the proposed outreach activities “will strengthen scientific research potential, not only during the funded period but for generations to come.”

Scientifically, there are still many more questions to be answered about the dynamics of intercellular bridges with such important consequences for so many species.

Behind the Bench Dr Amy Maddox

Michael Werner, PhD

Daniel B. Cortes, PhD

Kathryn Rehain-Bell

E: asm@unc.edu T: +1 919 843-3228 W: http://asmlab.web.unc.edu/

Research objectives Dr Maddox and her team are attempting to achieve the following: understand intercellular bridge composition, dynamics and regulation, capture live images of cell dynamics using different C. elegans tissues, and also, conduct extensive outreach activities within the University of North Carolina and beyond.

Funding • National Science Foundation (NSF) • NIH’s National Institutes of General Medical Sciences (NIGMS) Collaborators • Michael Werner, PhD

Q&A

What has been the most enjoyable and challenging aspect of your research?

It’s funny that you ask it like that, but indeed the most challenging aspects are the most enjoyable ones.

There are two classes of challenges: logistical and scientific. Logistically, it’s challenging to keep the lab funded and keep the people happy, but successes in both these realms have been incredibly rewarding. Scientifically, our forays into mathematical biology have really stretched me, but we’re making headway, mostly thanks to the fearlessness of trainees and generosity of collaborators. This diversification of our research program will doubtless propel us to new successes and enjoyment.

Were there any surprises for you due to working with such a diverse team?

I have actually been incredibly spoiled to recruit people not only with inner drive to study cell biology and development, but also with substantial training to tackle those questions.

In fact, all three of my current postgraduate lab members were previously

• Daniel B. Cortes, PhD • Kathryn Rehain-Bell The team acknowledges the valuable collaboration of Jian Liu (NHLBI), Francois Nedelec (EMBL), Adriana Dawes (Ohio State U), Wanda Strychalski (Case Western Reserve U), and many colleagues at UNC. Bio Team members have expertise in cell biology, high-resolution microscopy and computer-aided image analysis.

Amy Maddox was trained in North Carolina and San Diego and has served as faculty in Montreal, Canada. Michael E. Werner was trained in genetics and developmental cell biology in Austria trained to use C. elegans, our model animal of choice. Since joining my lab, Michael, Daniel and Katie have diversified from each other by pursuing the knowledge, technologies and skills necessary to advance their projects and their independent careers. I admire their independence and inner drive, and am grateful to be part of their journeys!

What has been the best thing about working with other faculty members?

I really appreciate how my faculty colleagues provide a stimulating, multidisciplinary environment, and express genuine interest in my scientific success.

I am also grateful for their support of my sometimes-wacky ideas to connect people in new and different ways.

They have been my “guinea pigs,” attending scientific speed dating events and randomised lunch dates, and they report enjoying these activities!

My most important colleague is my husband, who is a professor in the same department as I am. It is invaluable that we understand and can contribute to each other’s careers.

Do you expect that future work of this nature will be conducted with C. elegans?

Yes, we certainly have not exhausted

and Chicago. Daniel B. Cortes was trained in molecular and cell biology at the University of California at Davis.

Kathryn Rehain-Bell was trained in evolutionary developmental biology at the College of William and Mary in Virginia.

Contact Amy Shaub Maddox, PhD Assistant Professor of Biology University of North Carolina at Chapel Hill CB 3280 408 Fordham Hall Chapel Hill, NC 27599-3280 USA

the potential for this model animal to reveal fundamental principles of syncytial biology. Even as more cell biologists are discovering the elegance and accessibility of this tiny worm’s oogenic (egg-making) syncytial gonad, there are frontiers in the understanding the spermatogenic gonad, and in exploring the diversity of gonad architecture throughout nematode phylogeny and beyond.

How will a better understanding of intercellular bridge stability be useful for humans?

Since intercellular bridges are crucial for animal fertility, our discoveries will, in the long term, inform preventative or therapeutic medicine. Furthermore, findings of how multiple nuclei reside in one cell are relevant to other tissues (including heart muscle cells and certain fungi) with syncytial architecture.

Importantly, “basic” research aimed at defining the workings of the natural world will always enrich humanity by deepening our understanding in general, and we can almost never predict how and when our findings will directly serve human health and society.

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Biology ︱ Dr Chunqiang Li

Novel 3D microscope provides unprecedented moving images of biological processes Dr Chunqiang Li and his team of the University of Texas at El Paso have developed a novel three-dimensional (3D) optical microscope that uses a spectrally shaped pulse laser. Whilst most prior microscopes used scanning to achieve high speed 2D imaging, Dr Li’s approach obtains the z-position from a technique called ‘temporal focusing’ that use ‘diffraction’ rings and clever mathematics to infer the z-position.

3D defocusing imaging of microbes.

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r Li and his collaborative team have achieved a breakthrough in optical microscopy for 3D imaging without raster scanning laser beams. He expects to observe interaction between living cells at the microscopic scale, in three dimension (3D). Dr Li’s approach will permit cellular and molecular activities like protein motions, subcellular signalling, and viral infection to be examined and captured in real time.

The team have built on the work of others in the field who have sought to image the trajectory, in 3D, of tiny ‘particles’, molecules or cells for example, at a precision of a nanometre, or one billionth of a metre.

UNDERSTANDING 3D MICROSCOPY Earlier microscopes relied broadly on one of three techniques to capture 3D trajectory. The first uses a control mechanism called ‘feed-back’ to track the movement in all three dimensions of a single particle using the scanning of laser beams. The second technique involves utilising the shape change of an image illuminated with a focused point of light, called the ‘point spread function’ to calculate the z-position. The final method uses the diffraction of light of an image that has been defocused.

Unlike the previous method the resultant image contains concentric rings and the z-position can be determined with high accuracy from the radius and centre of the rings.

Fig. 1

Femtosecond laser

HWP

Prism pair

Polarizer

Mirror

Mirror

Pickoff mirror 3D stage Objective

DM1

L3

L2

AOM

L1

Sample

Grating DM2

BP1 Imaging lens Camera1

Camera2 BP2

AFG

Imaging lens

Schematic setup of the temporal focusing two-photon microscope.

THE TEAM AT EL PASO’S APPROACH Dr Li and his team have developed a microscope that is innovative in allowing the tracking of more than one particle using fluorescence. Fluorescent light is detected from the target ‘particles’ by an optical technique often used in microscopes called ‘two-photon excitation’. In this, the specimen is labelled to emit light in a particular colour when excited by a laser beam. Shaped ultrafast laser pulses illuminate the scene to be captured over a large volume, in microscope terms, of 100 µm, or millionths of a metre, each side. The sample is deliberately moved away from the focal plane of the microscope objective lens to allow defocused imaging, creating concentric rings caused. This approach, called ‘temporal focusing’ can not only obtain 2D images of the specimen, but also allows the z-position to be calculated from the defocused rings. Time-lapse images permit the progression of the particles being tracked to be captured and then viewed as a video or inspected individually.

BUILDING THE MICROSCOPE Dr Li’s microscope uses a laser as its illumination source, capable of delivering pulses in the femtosecond range (i.e., 10−15 or 0.000000000000001 seconds).

The ultrashort laser pulse is reduced in power using optical components

colours overlap spatially at the objective lens focal plane, they also overlap temporally to achieve short laser pulses.

The subjects being examined are fluorescently labelled so that they emit light at certain wavelength after being illuminated with the ultrafast laser pulse.

Dichroic beam splitters are used to separate the emission light from the excitation laser light. Images and videos are captured by cameras. Different types of subjects can be labelled with difference fluorescent agents to allow multiple channel imaging.

Dr Li demonstrated the power of his invention and was able to track the highspeed motion of Cafeteria roenbergensis as separate images that could be examined or combined into a 30 frames per second moving image before being compressed by passing the light through a pair of prisms. A special component called grating splits the light into separate colours and each colour ray passes through a lens for collimation, or make them travel in parallel paths like railway tracks.

A device called an ‘acousto-optic modulator’ can manipulate each separated laser colour ray before it passes through two further lenses that relay the laser pulse to the objective lens.

When all the laser light with different

WATCHING MARINE ZOOPLANKTON The operation of Dr Li and his team’s microscope was tested using a fastmoving marine zooplankton. They chose Cafeteria roenbergensis, a D-shaped biflagella single-celled organism. The flagella are hair-like appendages that permit the zooplankton to propel themselves around the oceans.

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At around 10μm across, they feed on bacteria and phytoplankton, are significant predators of marine bacteria, contributing to nutrient recycling, and are important in the marine food chain.

For the experiment, two colours of fluorescence agents were used to label Cafeteria roenbergensis: green and red, comprising tiny beads of 100nm in diameter.

The laboratory grew cultures of Cafeteria roenbergensis in culture medium, with Escherichia coli bacteria as their food source. For the experiment, small quantities of Cafeteria roenbergensis were extracted from the laboratory’s stock and transferred to a medium devoid of food. After four days of starvation, the diluted fluorescent beads were added to the culture. The zooplankton ingested the beads as food and could be used as targets in Dr Li’s team’s microscope to track their motion.

They were kept in a darkened place to preserve the fluorescence for the experiment.

Time-lapse images were taken with the two cameras, each camera capture one of the fluorescent colours, green or red. The techniques developed by Dr Li meant that each image showed the position of the organism in twodimensions and the defocused rings around each image inferred the third dimension. Using a mathematical model permitted the team to calculate the z-position of the two zooplankton being

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Dr Li’s novel approach could lead to exciting discoveries in biology and health as it waill be possible to examine the interaction between tiny organisms like viruses and their hosts tracked, denoted by green and red fluorescence, to add to the other two dimensions filmed.

Dr Li demonstrated the power of his invention and was able to track the highspeed motion of Cafeteria roenbergensis as separate frames that could be examined or combined into a 30 frames per second moving image.

IMPROVING THE MICROSCOPE The team have shown that it isn’t necessary to use a scanning laser to capture wide-field 3D images on a microscopic scale, instead they use two-photon fluorescence as a method and temporal focusing to reveal the z-position of a target.

Dr Li states that the current frame rate of 30 frames per second of their

microscope is not an upper limit, but is the result of the charged-couple-device cameras he used. He has proposed that his team’s approach could achieve millisecond resolution, and even higher speed operation is a possibility for the future.

Dr Li’s novel approach could lead to exciting discoveries in biology and health sciences as it may be possible to examine the interaction between tiny organisms like viruses and their hosts but it is not his only achievement. The microscope is not Dr Li’s only achievement from this research; he’s created complex mathematical models, utilising modelling software, to generate noise reduction algorithms and derive the z-position from the temporal focusing rings with high accuracy.

Behind the Bench Dr Chunqiang Li

E: cli@utep.edu T: +1 915 747 7537 W: http://science.utep.edu/photonics/ W: http://news.utep.edu/microscope-technology-will-enhance-array-of-studies/

Research Objectives Dr Li and his team have developed a high-speed temporal focusing twophoton fluorescence microscope for three-dimensional (3D) volumetric imaging without laser beam scanning.

It will give scientists in biology and chemistry a strong background in the development and use of innovative and contemporary instruments, significantly enhancing their ability to conduct cutting edge research. This will also provide training for next generation instrumentalists, who are currently postdoctoral researchers or graduate and undergraduate students.

Q&A

Your team’s approach to determining the z-position using the position of the rings in the defocused region of the image is novel. What drove you to select this mechanism?

Studying biological process in three dimensions is necessary as they happen in a 3D space. However, current optical microscopy can obtain 2D images/videos. The thirddimension information requires extra effort, such as scanning the object or lens. This mechanical scanning is the bottle neck for achieving high speed 3D imaging. Therefore, inventing a fast way to conduct 3D imaging is very useful in many research fields.

Defocused imaging can provide not only 2D information, but also the third dimension information from the level of defocusing. Therefore, it is my vision to fully utilise the information contained in such imaging setup.

The acousto-optic modulator is an integral part of your microscope

Funding National Science Foundation (NSF) Collaborators • Wei Qian (Electrical and Computer Engineering) • Jorge Gardea-Torresdey (Chemistry) • Chuan Xiao (Chemistry) • Kyung-An Han (Biological Sciences) Bio Dr Chunqiang Li received a Doctor of Philosophy in Electrical Engineering from Princeton University in 2006 and, from 2006 until 2010, he was a Postdoctoral Fellow at Wellman Centre

and allows you some variability in the design. How would this feature in a commercialised microscope?

This acousto-optic modulator is used to control the spectrum of our femtosecond laser. It has been used in some products in steering laser beams only. Our method explores its novel use of shaping the laser pulse in spectral domain. The incorporation of this device in future commercial microscope will expand our ability to have better control of laser beams, which opens many more potential applications.

In your experiments, you used two frequencies of light to track two particles simultaneously. How could this be achieved with many particles?

In many particles, it is necessary to develop multi-frequency tracking, i.e., each particle is labelled with one-colour label, and our microscope can visualise them simultaneously.

Can you explain how the microscope would operate if tracking the infection of a cell with a virus?

The virus is stained with one dye at one colour; and the cell is stained

for Photo-medicine in Massachusetts at Harvard Medical School. Since 2010, Dr Li has worked within the Department of Physics of the University of Texas at El Paso, leading the Bio-photonics Laboratory.

Contact Chunqiang Li, PhD Associate Professor Physics Department University of Texas at El Paso El Paso, TX 79968 USA

with another dye at another colour.

Both species are visualised with our microscope, and the laser can excite both dyes simultaneously. Two CCD cameras are used, one to detect the first dye, and the other one to detect the second dye. Therefore, there are two detection channels, one for each species. The final image/video is a mix of signals from both channels.

You mentioned the limitations caused by the cameras in use in the microscope, particularly the frame rate. You also suggested some ways around the camera limitations. What ideally would you like to achieve, and how?

There is a trade-off between imaging speed and signal-to-noise ratio in detecting such week fluorescence signals. If the fluorescent signal is strong, the imaging speed can be higher. Therefore, I would like to use bright and robust fluorescence molecules as the label to reduce the image acquisition time. The ideal case would be to achieve sub millisecond time resolution to study fast molecular dynamics in live animals.

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Thought Leader

CSMB: Ensuring a strong future for science and scientists As scientists make fast and concerted advancements in life-saving technologies, the squeeze on funding could threaten their momentum. That’s where Dr Philip Hieter, President of the Canadian Society for Molecular Biosciences (CSMB), comes in. To get the ‘biggest bang for their buck’, the CSMB is leveraging its collective knowledge and talents by focusing on the most pressing issues facing fundamental research, like funding. Through advocacy and education, they aim to influence not only present and future scientists, but decision makers in the Canadian government and the world.

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n 1957 the Canadian Biochemical Society (CBS) was first conceived. After merging with the Canadian Society of Cellular and Molecular Biology in 1995 and the Genetics Society of Canada in 2010, it renamed itself the Canadian Society for Molecular Biosciences (CSMB) in 2011. While the society’s name and make-up have changed over the years, its basic premise has been to support science.

And while science is making leaps and bounds on its way to combatting major diseases, it’s sometimes hard to believe, given the state of funding for fundamental research in Canada, after a crippling decrease in financial support over the past decade and a general lack of understanding of the system’s needs.

Fortunately, CSMB and other organisations have been working on ways to ensure that scientists can do what they do best, make amazing discoveries. CSMB is a relatively small organisation with a finite budget and members who realise that to have the biggest impact they need to broadcast the exciting possibilities and their importance both to government and to the general population. They believe that with advocacy and education, the necessary funding and support will follow. Dr Philip Hieter, President of CSMB, spoke with us at Research Outreach about the society and his excitement about its future.

Hi Philip! Could you tell us what your role involves as the President of the Canadian Society for Molecular Biosciences (CSMB)?

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During my time as CSMB President, I will help guide the CSMB board in identifying the pressing issues confronting the basic scientific research community, devising plans of action, and working to enhance the support Phil Hieter looking at yeast colonies growing on a Petri plate.

of science. I brought with me previous experience in my role as President of the Genetics Society of America (GSA) a 5000-6000-member organisation based in Washington, DC. The GSA, as its name implies, is primarily focused on

The Michael Smith Laboratories Building at the University of British Columbia, where scientists conduct fundamental research in molecular genetics, bio-process engineering, bioinformatics, and genomics.

There is a beauty in discovery itself. I think society is gratified by trying to understand nature, understanding the world around us the community of genetics researchers, whereas, CSMB has a broader mandate A key immediate issue is promoting a re-investment by government into the scientific funding envelope that is necessary to support science research and teaching in Canada at an internationally competitive level.

Adequate support of fundamental discovery research is critical to the realisation of long term societal benefits in health, agriculture, climate, and industry. We’ll also address standards and practices that ensure success at all stages of scientific career development.

The CSMB also strongly believes that a robust, research-based university

system, which supports cutting edge technology and approaches, is a critical context for training the next generation of innovators, and for teaching science more broadly.

Can you tell us more about the CSMB’s heritage and background?

CSMB is an amalgam and evolution of three more focused societies. What began as the Canadian Biochemical Society, became Biochemistry and Molecular Biology before merging with the Cell and Molecular Biology Society, and finally with the Genetics Society of Canada in 2010. The unification of biochemistry, cell biology, and genetics helped to concentrate energies and

efforts – and provided critical mass. I became actively involved about the time the merged societies became the CSMB. How do you advocate the progression of molecular bioscience? What is the value of basic science to society?

Why does it need to be supported by governments?

Every basic discovery has the potential of affecting major applications in health, agriculture, and industry. This is often a long-term progression, and federal governments are the only bodies that take on that timeframe without shortterm economic return. Since we’re talking about scientific research that leads to solutions in health treatment

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After DNA replication, chromosomes (light blue) condense and undergo a series of coordinated movements to achieve bipolar attachment of sister centromeres on the mitotic spindle (green) at metaphase (A and B). Once properly attached, the replicated sister chromosomes are coordinately pulled to opposite poles during anaphase (C). Images courtesy of Dr Conly L Rieder (conlyrieder@hotmail.com). The Hieter laboratory studies chromosome biology in yeast and human cancer.

and management, as well as other major problems worldwide, it’s vital.

Under a previous administration, the long-term recurring money available for reaearch grants to basic scientists declined significantly. This has caused a devastating shortfall in the operating budgets that fuel the day-to-day operations of individual research laboratories. It dramatically crippled our pipeline for discovery and training at the early stages in the broad continuum of research from basic to applied, that ultimately leads to direct applications. If a government pulls funding from early training and fundamental research, it pays the price five, ten, twenty years down the road.

That’s why we’re so excited about the recommendations made in the Trudeau government’s “Fundamental Review of Science,” (April 2017), also known as the Naylor report. It recommends a large reinvestment in research operating grants (back to 2007 levels), better coordination across the federal funding agencies, and financial support of open competitions for early and middle career

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scientists. Using our ‘inclusive’ advocacy, education programmes, website and social media, we plan on broadcasting this information to our members, the broader scientific community, advocacy organisations (lobbyists), and people at every level in our community, including decision-makers in government.

There is beauty in discovery itself. I think society is gratified by trying to understand nature, understanding the world around us. But there are practical benefits that are quite large and more than justify the investment by governments. This truth and awareness should help drive longterm funding and support.

What are the main research interests and key areas of focus current at the CSMB? From a scientific standpoint, we try to understand the basic mechanisms of cells. Our members include researchers in genetics, cell biology and

biochemistry, the perfect combination for focusing on a gene or a pathway to understand its function. Genetics – finds mutants that define genes and manipulates the genome to understand the consequences gene disruption in cells. Biochemistry – helps us understand how gene products work at a structural and molecular level. Cell biological methods – akin to imaging, explore the dynamics of gene function in cells. The three together accurately define the functions of genes.

Our last two presidents, Christian Baron (University of Montreal) and Kristin Baetz (University of Ottawa) helped revitalise the CSMB. Christian upgraded our website, our social media presence and rallied the Board to achieve defined goals in advocacy and training. Kristin staged our 150th Anniversary of Science meeting in Ottawa to encourage interaction with MPs and instituted a

Thought Leader dual-track speaker programme with lots of engaging crossover. Kristin also worked tirelessly in advocacy efforts through direct interaction with government officals in Ottawa.

We will keep the momentum going and continue to share the big picture of what we do and how important it is to the world. If we can’t support our scientists and be internationally competitive, how

CSMB explores serious issues and makes strong recommendations in the form of petitions. It can be difficult to measure the impact of our petitions, but we imagine they do have an effect. We’re proud of our role in arguing against the introduction of a detrimental grant review process at the Canadian Institutes of Health Research (CIHR) that was eventually overturned. Thousands of people signed

DNA sequencing of whole genomes with our access to large numbers of people with genetic variants that cause disease, we will be able to determine, for example, the particular gene mutations that cause each of the various sub-types of cancer and be able to rationally prescribe effective drugs, based on an understanding of the specific biological mechanisms at play. These technological

A win for one scientist is a win for the entire scientific community and the world will we develop our next generation of scientists and solve the problems we face? We simply can’t afford to allow them to fail.

CSMB offer numerous awards to both researchers and students, including the New Investigator Award, the 2016 Video Challenge and the Graduate Student PDF Poster Competition.

What are the benefits of having these award schemes in place?

Every year, scientists make groundbreaking discoveries and we feel that recognising and celebrating their excellence is invaluable, plus it is also great fun! It’s important to remember that a win for one scientist is a win for the entire scientific community and the world.

Indeed, it is naturally quite a motivation and encouragement to young scientists.

These awards also increase our visibility and credibility which may help us to influence the approval of ideas and policies to support science, for example, as in the recommended in the Naylor Report. This historic document addresses the state of science in Canada and where it needs to go – a rigorous accounting, and a clear roadmap for the future. The CSMB and Canadian scientific community is also encouraged by the installation of the Canadian Minister of Science, Kirsty Duncan, and the Chief Science Advisor, Mona Nemer.

The society runs various petitions seeking public support. What impact do these petitions have and what topics are you currently petitioning for or against?

our petitions, and in coordination with the efforts of many others, we saw the return of the gold standard face-to-face peer review mechanism for determining the best grants.

In my own laboratory, CIHR grants made possible our basic studies of genes and pathways in yeast as a model system for understanding the corresponding human genes that are mutated and cause the progression of human cancer. Even though we do basic studies on chromosome biology in yeast, there’s a strong relevance of what we discover in yeast to an understanding of cancer biology. My research has been funded by federal health research agencies over the past 30 years, and we have close interactions with both basic and clinician scientists.

CSMB petitions have advocated for a balance between fundamental and applied research to ensure a healthy sustainable research ecosystem vital to fundamental discoveries and innovation.

These advocacy efforts will continue to be a top priority for the society in the years to come.

Finally, where do you see molecular biosciences going over the next ten years or so? Are there any areas that you are particularly excited about?

We’re all excited about CRISPR, molecular biosciences’ new method for making very specific directed changes to the genome of essentially any organism.

This new ‘precision genetics’ technology has broad applications in all areas of boiology. Coupling next-generation

breakthroughs are not just going to impact major diseases like cancer.

Hundreds of new disease genes that cause rare human diseases are being discovered every year. These dicoveries provide a molecular diagnostic for DNA testing for them, and open up research avenues for developing novel treatments. CSMB will continue to do everything we can to help scientists keep making phenomenal strides.

• If you would like to find out more information and the work of the CSMB, please visit their website at www. csmb-scbm.ca/index.aspx.

Contact Canadian Society for Molecular Biosciences c/o Rofail Conference and Management Services (RCMS) 17 Dossetter Way Ottawa, ON K1G 4S3 Canada E: contact@csmb-scbm.ca T: +1 (613) 421 7229 W: http://www.csmb-scbm.ca/ index.aspx

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Biology ︱ Dr Hilary Swain

Florida’s Archbold Biological Station gives online access to unusual natural history collection The Archbold Biological Station, a world-renowned ecological field station based in Florida, USA, is uploading its natural history collection onto the Internet for the first time. The diverse collection, containing 270,000 specimens of more than 10,000 species will provide researchers and students around the world, with access to this rich source of ecological data. This highly collaborative project, which involves making data and images of thousands of biological specimens available online, is funded by NSF and is being led by Dr Hilary Swain and Dr Mark Deyrup.

F

ounded in 1941 by Richard Archbold, a biological explorer, the Archbold Biological Station (Archbold) is an internationally renowned, not-for-profit biological field station located in central Florida. Archbold manages nearly 20,000 acres of land, is dedicated to research and conservation programs, and conducts scientific studies at more than 50 locations throughout the headwaters of the Everglades – a 2.6-million-acre watershed in southcentral Florida encompassing those lands and waters that drain south to the Everglades and onto the coasts. The region is one of the most important biodiversity “hot spots” in North America. The endangered Florida scrub is of special interest, being a harsh and stressful habitat that is home to many plants and animals found nowhere else on Earth. Through its research, education and outreach programs, Archbold aims to conserve this biodiversity, and maintain vital ecosystem processes that support Florida’s natural environment and its people. The research addresses many of the most important issues facing our world today, such as: climate change, species and land conservation, water quality, and sustainable food production.

The Station is also home to the Archbold Natural History collection – a diverse record of life taken largely from the habitats of the ancient Lake Wales Ridge, a 100-mile-long north-south sand ridge running up the interior of the Florida Peninsula. The 75-yearold collection has grown through the years, and currently contains more than

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Future museum specimens being wrangled from a dead oak; the ecological data will be included on the specimen labels.

Photo by Dustin Angell.

270,000 specimens and over 10,000 different species. Although smaller than the vast collections at some universities and at large museums, the Archbold collection is valuable in that it is representative of in-depth and very rich regional collections, with a nearly complete local diversity rarely captured elsewhere. “We all believe that natural areas teem with biodiversity,” says Archbold entomologist Dr Mark Deyrup. “Here is proof. For example, more than 1,500 species of beetles live on this single site.” Overall, it is one of the largest collections for any North American site and includes a wide variety of arthropods, plants, mammals, birds, fish, reptiles and amphibians. It also houses specimens of threatened and endangered plants and animals, giving scientists a unique opportunity to study these rare species.

And it includes specimens of many newly arriving species, some of which are invasive, thus tracking changes in Florida’s diversity over time. Now, with National Science Foundation (NSF) Collections in Support of Biological Research (CSBR) funding, Dr Hilary Swain and colleagues, aim to share the

The chemical ecology of the Bella Moth (Utetheisa bella) has been studied at Archbold for many years. Specimens in the collection serve as archived vouchers, tying together field research and publications.

Natural History collection with a much wider audience, putting Archbold on the global stage.

DIGITAL COLLECTION During the last couple of years, a team of scientists led by Dr Swain

at the Station have been diligently photographing specimens, and adding label information of the flora and fauna into a searchable database. Significant portions of the collection have been databased and now, for the first time, are being made available online

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Left: Museum specimens can reflect major ecological events. Tiny beetles were found in the hidden rotten heart of oaks split by Hurricane Irma on September 10, 2017. Photo by Dustin Angell.

Above: Tiny beetles (sesame seed for reference) found in rotten oak include six species never seen before in 75 years of beetle study at Archbold.

The online arthropod collection in particular is absolutely outstanding, as the Archbold Natural History collection is best known for its pinned collection of 250,000 insects

In particular, the online arthropod collection is outstanding, as the Archbold Natural History collection is best known for its pinned collection of 250,000 insects, including 137,450 preserved specimens of ants; this encompasses the largest collection of Florida ants in the world. Currently a total of 9,718 ant specimen records – representing 236 species of ants – have been uploaded online to SCAN, including detailed drawings completed

by Dr Deyrup (see example opposite).

Over the years, the arthropod collection has served as the basis for many scientific papers, contributing to numerous studies requiring insect identification, and was the main source for the Dr Deyrup’s 2016 book Ants of Florida. The book is a natural history and identification guide for all 239 species of Florida ants.

through several national collection portals. This will allow scientists, school children and the general public from all over the world access to this unique collection. To do this, Archbold researchers are working with partner organisation Integrated Digitized Biocollections (iDigBio), an organisation funded by NSF that is making data and images of millions of biological specimens available in electronic format for the research community, government agencies, students, educators, and the general public.

The collection also includes specimens of insects that were captured feeding on nectar and pollen of local flowers.

This record of ‘flower-insect visitor interactions’ was begun in 1983. It includes more than 5,000 specimens of bees and wasps, 1,500 flies, 200 Lepidoptera (butterflies and moths) and 500 beetles, with labels that record the species of flower being visited by the insect, the location, and the date of capture. So far, a total of 10,868 flower-insect visitor specimens have been uploaded to the online database

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Assisted by iDigBio, Archbold staff has currently uploaded approximately 30,000 records of plants, birds and arthropods to online portals hosted by the Symbiota Virtual Biota software package. To date these include 24,000 records of pinned insects available on the Symbiota Collection of Arthropods Network (SCAN), 1,474 records of bird skins available on the Consortium of Small Vertebrate Collections (CSVcoll), and 4,795 herbarium records of pressed plants available on the North American Network of Small Herbaria (NANSH).

SCAN, representing a total of 902 insect species. This ecological dataset is an important resource for biologists, as it can answer questions like: Which bees visit which flowers and when? At the community level, this information is now available to allow researchers to model the stunningly complex network of relationships among flowers and insects at a single site.

WIDENING ACCESS The online collection will be a huge resource for scientists around the world, providing them with easy access to the data they need to conduct their research. Plant specimens, for example have been used by scientists studying flowering times, which can be affected by climate change. The vertebrate collection provides a vital resource for scientists wishing to study the variation, growth patterns, life histories, and population dynamics of animals. The database will also be of huge interest to schools and children.

Archbold has been dedicated to

The lands and waters that form the headwaters of the Evergaldes are notable for extensive natural areas that have never been cultivated or strongly disturbed. Here Archbold scientists study species of plants and animals that have been in residence for thousands of years. Figure by Archbold Biological Station.

Temnothorax smithi. Multiple specimens in collections allow scientists to determine which features are characteristic of a species and which are variable. After examining many specimens of this ant species in the Archbold collection Mark Deyrup drew this generalised diagnostic image.

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educational outreach for many years. It shares its knowledge and habitats with students of many ages, from age seven through to adult learners. It provides environmental education resources to local schools, as well as access to the site for school field visits and summer camps. It hosts an estimated 2,000 3rd-5th grade and middle school children annually, and nearly 50,000 school visitors have benefitted from its outreach program in the last few years.

Undergraduate and graduate university students regularly visit the Station and use Archbold research to conduct field courses or carry out independent field studies. Archbold also provides unique 6-12-month internships, longer than most internships offered by other organisations, where students receive

The 75-year-old collection has been expanded over many years, and currently contains more than 270,000 specimens and over 10,000 different species training, participate in research projects and, most importantly, are required to conduct independent research projects of their own design.

The online Archbold collection will make the Station and its work even more accessible to learners and the younger generation. Not only will it be a source of ideas and specimen data for children and undergraduates/graduates from around the world, it will fuel the

minds of the younger generation.

“This is a giant trove of information for them,” says one Archbold researcher.

“Advances in computer analyses and graphics will allow the next generation of scientists to swim joyously in floods of data that would drown members of our own generation.” Scientists hope that learning to understand and value complexities of the natural world will inspire conservation of rare habitats such as Florida scrub.

•Data collection websites to visit: http://symbiota.org (Symbiota Virtual Biota software package), http://symbiota4.acis.ufl.edu/ scan/portal/ (Symbiota Collection of Arthropods Network (SCAN)), http://csvcoll.org/portal/ (Consortium of Small Vertebrate Collections (CSVcoll)), and http://www.nansh.org/portal/ (North American Network of Small Herbaria (NANSH)).

Behind the Bench Dr Hilary Swain

E: hswain@archbold-station.org T: +1 863 465 2571 (Use EXTN 251) W: http://www.archbold-station.org

Research Objectives Dr Swain, Dr Deyrup and their collaborators’ aim is to increase digital access to the Archbold Collection, enabling studies of biodiversity in ways that would be difficult to replicate elsewhere, and enhancing its conservation value as a critical repository for species of the globally threatened Florida scrub. In addition, this project aims to offer students and research interns more opportunities for use of the collection as a source for research ideas and study specimens, as well as provide new mentoring for this next generation of field station users in the importance of collections.

Funding National Science Foundation (NSF)

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Collaborators Co-Project Investigators: Dr Mark Deyrup, Dr Reed Bowman, Dr Eric S Menges, Dr Betsie B Rothermel, Stephanie Leon and Stephanie Koontz.

Research Interns: Gabrielle LaTora, Carly Tolle, Ryan Huether, Dylan Ricke, Katherine Beigel and Trevor Young.

Graduate Students: Tram Nguyen and Young Ha Suh from the Cornell Lab of Ornithology who aided in the digitisation of bird specimens.

Nancy Deyrup is a volunteer in the entomology program who singlehandedly digitised the majority of the insect-flower database. Dr Butch Norden, volunteer, organised the amphibian and reptile collection for digitisation.

Bio Hilary Swain has been the Executive Director of Archbold since 1995, directing activities at Archbold Biological Station and the MacArthur Agro-ecology Research Center (MAERC). She works with a staff of 50 involved in long-term research, environmental monitoring, science education for students and the public.

Contact Hilary M. Swain, PhD, Executive Director Archbold Biological Station 123 Main Dr Venus, FL, 33960 USA

Q&A

This collections grant is an amazing project to be involved with! What are the major benefits this grant will bring?

Swain: First and foremost, the major benefit of the grant is to enable and enhance more scientific research. As anticipated by our funder, the National Science Foundation, we already see that having Archbold’s collection online is shining a light on these data for scientists around the world, not just those who visit Archbold. Whether scientific interests lie in identifying new species, cataloguing biodiversity, highlighting the arrival of new species, noting changes in the timing of flowering, or building complex networking analyses for insect flower visitors, this collection is a treasure trove of ecological information just waiting for scientific discovery and synthesis. Archbold is one of many biological field stations around the world that are important to scientists because although their collections are relatively small, they are extremely important representatives of regional biodiversity.

Leon: One major benefit of this grant is demonstrating the importance of natural history collections, and how they can be used outside of a museum setting. More importantly, the fruits of this grant can demonstrate how a wellcurated, regional, on-site collection can provide useful data not only for taxonomy and systematics, but also for ecology and conservation.

Deyrup: Natural history collections are currently cascading onto the Internet, escaping from their reputation as mouldering mortuaries to present their true aspect as enormous and dynamic sources of original information.

Why is it important that a record of life in the Florida Scrub habitat is made and preserved?

Swain: Since scientists first visited the Florida scrub and started collecting specimens of its plants and animals, it has always been recognised as one of the most threatened ecosystems in the USA. Like nowhere else on Earth, there has always been a scientific race to ensure we capture and catalogue all its precious life and unique adaptations.

Deyrup: As we consider our own survival in a world of increasing

Q&A with Dr Hilary Swain, Dr Mark Deyrup and Stephanie Leon environmental stress, we could benefit from examining the information and morphology associated with organisms inured to a land of infertile sand, and swept with fires, floods and droughts.

Leon: The Florida scrub is a unique habitat with diverse flora and fauna.

It is important to show how such a harsh habitat can support so much biodiversity. This natural history record will also show why conservation of natural areas is important.

How will researchers use this data?

What scientific questions could be answered?

Leon: Our data is useful at many levels. It can inform scientists who are interested in the distribution of a particular species, it provides a record of rare and endangered species, it can aid taxonomic revisions and descriptions of new species, and it also provides useful ecological information on plant-insect interactions, primarily pollination. Those interested in pollination ecology can find our “flower visitor” dataset online, download it, and use it to create intricate networks to illustrate the complexity of these relationships.

Deyrup: As data and images of expertly prepared natural history collections go online they become a kind of universally accessible museum – with the added feature that every scientist can rearrange the “exhibits” to further their individual goals.

How will school children benefit from accessing the database?

Swain: Children are natural observers.

Their curiosity especially draws them into the spectacular photography of our plants, birds, and the bugs in the online databases. Their faces watch with awe as a small bug looms large on the screen: they see the spectacular colours and intriguing shapes. Here lies inspiration for a future structural engineer, computer modeller, fashion designer, or budding ecologist.

Hopefully it will also draw them into the natural world where they can use their sharp and laser-focused eyes to be further intrigued by nature.

Leon: Children will learn about the diversity found in the Florida scrub. It might encourage exploration, not only of the scrub, but of different natural

habitats. It might also supplement their learning, research projects, and general interest of biodiversity. Many children that visit Archbold are given tours of the Arthropod collection; they learn about the vast diversity of insect species collected at Archbold. This might entice some to start their own personal collections.

How will the archive further conservation efforts in Florida?

Swain: Biodiversity is in flux worldwide, and nowhere more than in rapidly changing and threatened ecosystems like the Florida scrub. Apart from the obvious conservation question, “What is out there”, we can use a collection like Archbold’s to inform conservation in many different ways.

How is biodiversity responding to changing environmental conditions over time? What new and potentially invasive species are arriving? How does conservation management, such as prescribed fire, benefit species of concern? Are we inadvertently overlooking ecosystem components that are more critical for conservation than we realise, such as Mark Deyrup’s long-term fascination with documenting all the insects supported by recently burned, dying, dead, and decomposing wood.

Leon: Our goal is to demonstrate that natural history collections can function jointly with ecological research and conservation efforts. In order to do this, we have to showcase the biodiversity of this unique habitat, and how this biodiversity might be changing over time. Rare, endangered plants are an example of how we can use historical records to track changes in their distributions, and also track the history of invasives, to aid in conservation and/ or restoration efforts.

Deyrup: Florida has an exploding human population and amazing numbers of invasive species, combined with impending effects of climate change on a vulnerable landscape.

Collection records give a baseline of species diversity and distribution, allowing targeting of conservation.

As the richness of Florida’s natural heritage becomes easily visible through natural history archives it should inspire the next generation of conservationists.

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Biology ︹ Dr Imara Perera

Communicating the language of plants through inositol phosphates Organisms require a variety of signalling pathways to communicate with and respond to their environment.

Components such as signalling molecules function as the communicators of cellular language. Dr Imara Perera and her colleagues are investigating one group of signalling molecules, inositol phosphates and their derivatives, to understand its role in plant communication.

A

ll organisms interact with and respond to their internal and external environmental conditions. Acting on these responses requires a language comprised of cellular words which interpret and relay information within the organism.

One category of words, or signalling molecules, are the myo-inositol phosphates (InsPs) which have been implicated in a variety of processes within the cell. And much like words, these molecules vary in the number and position of letters (in this case phosphates) with each combination meaning different things in eukaryotic cellular communication.

Examples of data collected in studies on inositol phosphates and pyrophosphates. Arabidopsis plants growing in a 96-well plate can be grown under low energy conditions and analysed for inositol phosphate changes in response to changing energy conditions.

One particular group of these signalling molecules are characterised by diphosphate (PP) or triphosphate (PPP) chains. These inositol pyrophosphates (PPx-InsPs) have been implicated in cellular metabolic processes given the similarity of its structure to a molecule called adenosine triphosphate (ATP), sometimes referred to as the energy currency of cells. Dr Imara Perera, Dr Glenda Gillaspy and colleagues: Dr Joel Ducoste and Dr Cranos Williams are interested in the function of this group of signalling molecules in eukaryotic organisms such as plants.

INOSITOL HEXAKISPHOSPHATE – A PRECURSOR TO INOSITOL PYROPHOSPHATES Inositol phosphates comprise an inositol ring and varying numbers and positions of phosphates attached to it, each communicating unique messages in the cell. A fully phosphorylated form is inositol hexakisphosphate (InsP6), a potential signalling molecule and a precursor to the aforementioned group of inositol phostphates PPx-InsPs that Perera and colleagues are studying.

Inositol hexakisphosphate acts as a storage compound of inositol, phosphorus and minerals and is found in large quantities in seeds. This large quantity of InsP6 is later hydrolysed during germination and has led researchers to believe that as a PPxInsPs precursor, it is likely that plants also synthesise this unique group of signalling molecules. However, very little attention has been paid to the presence of PPx-InsPs in plants which is why Dr Imera Perera and colleagues have stepped in. This coupled with PPx-InsPs role in ATP homeostasis make them an interesting group of signalling molecules to study.

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Schematic showing how Arabidopsis seedlings are grown to facilitate labelling of inositol phosphates, that can be detected in the laboratory with high performance liquid chromatography and radioisotope detection.

INOSITOL PYROPHOSPHATE SYNTHESIS AND FUNCTIONALITY One avenue of research is understanding the enzymes which synthesise PPx-InsPs molecules.

In fact, synthesis of InsPs and PPxInsPs molecules is carried out by two evolutionary-conserved InsPs kinases.

These kinases catalyse the addition of pyrophosphates at distinct positions on the inositol ring. The first class are delineated IP6Ks (but KCS1 in yeast and absent in plants) and the second class are VIPs (or PPIP5K in humans) which is conserved across eukaryotes. It is these InsPs kinases which phosphorylate InsP6 produces derivatives such as inositol pyrophosphates InsP7 and InsP8.

Previous research has established InsP7 and InsP8 as important molecules in metabolic programming as their energyrich pyrophosphate moieties make them comparable to ATP. However, they have not been widely studied in plants.

By tagging important proteins with fluorescent markers, the authors can visualise where in the cell enzymes that break down inositol pyrophosphates reside.

The authors utilise plant mutants that cannot make normal amounts of inositol phosphates (on right), and compare responses to those from native, or so-called wildtype plants (on left).

Using a variety of techniques, Dr Perera and her colleagues identified the presence of these derivatives in higher plant tissue. This was further supported by the fact that Arabidopsis and maize seeds with a mutation in the InsP6 transporter protein produced plants

These inositol pyrophosphates (PPx-InsPs) have been implicated in cellular metabolic processes given the similarity of its structure to a molecule called adenosine triphosphate (ATP), sometimes referred to as the energy currency of cells

with elevated levels of InsP7 and InsP8.

One hypothesis for this is that a block in transportation of InsP6 provides a larger pool for VIP kinases to access and thus produce InsP7 and InsP8. They also identified two similar Arabidopsis genes, AtVIP1 and AtVIP2 which functioned to restore InsP7 synthesis in yeast, clearly demonstrating that plants can synthesis pyrophosphates. What’s more is researchers found that AtVIP1 and AtVIP2 were differentially expressed in different plant tissue, alluding to distinct functional roles.

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An example of radio-labelled inositol phosphate separation allows the authors to quantify these important signalling molecules.

But what function do inositol pyrophosphates play in plant signalling pathways and processes? Drs Perera, Gillaspy, Ducoste and Williams are exploring functions of PPx-InsPs, drawing from a variety of studies. New work by others has linked PPX-InsPs to phosphate sensing (Wild et al., 2017; Puga et al 2017; Jung et al 2018), a key process that allows plants to regulate growth in sync with phosphate, one of the most important nutrients in the soil environment.

Previous studies have found that InsP7 levels change in some non-plant organisms in response to limiting phosphate (Azevedo and Saiardi 2016).

PPx-InsPs have also been implicated in Jasmonic acid signalling and plant defense responses (Laha et al., 2015). As mentioned above, the similarity of PPxInsPs to the energy currency molecule ATP implies that they are involved in processes involving energy homeostasis, a crucial function in all organisms.

Phosphorous in undigested InsP6 seeds from non-ruminant animals has led to pollution of watersheds across the United States methods, such as kinetic modelling, to predict how InsP synthesis is modulated and identify key regulatory steps in the pathway. This system is based off differential equations which represent major reactions involved in the PPx-InsP pathway. By simulating these reactions, they can compare these to observations seen in mutants where loss of function occurs. By utilising this methodology,

FURTHER AVENUES However, there are still unanswered and unexplored trajectories in this line of research. Genetics and biochemical approaches aside, Dr Perera and colleagues hope to use computational

Inositol Pyrophosphate Synthesis plasma membrane

PtdIns

PtdInsP2

PtdInsP

PLC

myo-Inositol

Lipid Dependent Pathway

InsP3 IPK2

5PP-Ins(1,2,3,4,6)P5

InsP5 IPK1 P

P

P P

P

P P

InsP6

Storage

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P

P P

1,5PP-Ins(2,3,4,6)P4

VIP1, VIP2

P P

VIP1, VIP2

MRP5

P

P

InsP7

P

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PtdOH

IPK2

Lipid Independent Pathway

P

DAG

P

P

P P P

P

P

P P

P P

P P

P

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1PP-Ins(2,3,4,5,6)P5

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InsP8

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researchers can understand the potential behaviour of this pathway as well as key components in the signalling pathway.

Given the genetic conservation of proteins implicated in the InsPs signalling pathway, further research by Dr Perera and her colleagues will allow these findings to form the basis of insights into other eukaryotes, such as humans. In other words, understanding how InsPs play a role in signalling in plants, may be used to understand this pathway in other multicellular organisms.

On a bigger scale, research on this signalling pathway has the potential to understand plant metabolism in the context of agricultural production.

In addition to this, phosphorous in undigested InsP6 seeds from nonruminant animals has led to pollution of watersheds across the United States.

Better understanding of the signalling pathway may produce a solution to this toxicity issue. Research of this kind will therefore allow for the understanding of signalling pathways at the cellular level but also the environmental level with which these molecules interact.

Behind the Bench Dr Imara Perera

Dr Gillaspy

E: iperera@ncsu.edu T: +1 919 5 5 6985 W: http://pmb.cals.ncsu.edu/people/people-table/dr-imara-perera/

Research Objectives Dr Perera and her colleagues’ aim with this project is to understand how inositol phosphates are used to sense the energy and nutrient status of plant cells, and how they signal this status so that the plant can use this information.

Funding National Science Foundation (NSF) Collaborators • Dr Glenda Gillaspy (Virginia Tech) • Dr Pablo Sobrado (Virginia Tech)

Q&A

How reliable are comparisons to similar pathways in eukaryotic organisms?

Although pathways are conserved between eukaryotic organisms, there are significant changes between plants and animals, in specific components of the InsP pathway, as well as how they are generated and regulated.

For example, one key difference in inositol pyrophosphate synthesis is that plants only contain the VIP class of enzymes whereas animals and yeast have two different types of enzymes (IP6K and VIPs). Additionally, InsP6 is highly abundant and the major phosphate store in plant seeds, and no analogous situation occurs in most other eukaryotes.

Why can’t some animals digest InsP6 in seeds? And what are the repercussions of phosphate pollution?

The phosphate bound up in the InsP6 molecule is most easily liberated by specific enzymes present in plants, ruminant animals, and microbes in our

• Dr Joel Ducoste (North Carolina State University) • Dr Cranos Williams (North Carolina State University) Bio Dr Perera is a Research Professor in the Department of Plant and Microbial Biology at North Carolina State University. She received her PhD in Plant Biology from the University of Illinois at Urbana-Champaign.

environment. Non-ruminant animals do not possess these enzymes in their gut, and thus they cannot breakdown InsP6. As a result, undigested InsP6 is excreted into the environment. This along with excess unused phosphate from fertiliser ends up in agricultural run-off and causes pollution of fresh water sources. Experimental approaches to reclaim or phosphate from run-off are a focus of several interdisciplinary research teams.

Is it likely that VIP genes vary amongst different plant species?

To our knowledge, VIP genes are conserved in their DNA sequence across different plant species. As small differences in VIP gene sequences are present, we cannot rule out the possibility that some plants encode VIP enzymes with different properties. Thus, it is possible that there are species-specific differences as well as gene duplication.

What is kinetic modelling and how can it reveal key components in the PPxInsP signalling pathways?

Kinetic models of signalling networks are a mechanistic description of the key reactions that take place within the cell. Scientists can build kinetic models

Dr Gillaspy is Professor and Head of Biochemistry at Virginia Tech. Glenda and Imara share common interests and a productive collaboration on inositol phosphate signaling in plants.

Contact Dr Imara Y. Perera Research Professor Department of Plant Biology North Carolina State University Campus Box 7612 Raleigh NC 27695 USA

utilising known biochemical and biological data, and by estimating key parameters. Our initial PPx-InsP model uses biochemical data from a variety of eukaryotic enzymes and makes use of observed changes in PPx-InsPs in plant mutants. By fitting the observed data to the model, we can predict novel regulatory components.

How did you create InsP7 mutants in yeast?

We did not create these mutants but were able to make use of mutants created by other researchers in the field. The VIP mutants used in our study were obtained from Dr John York of Vanderbilt University. The York laboratory created these mutants by inserting DNA to disrupt key genes in PPx-InsP synthesis and degradation pathways. We introduced the plant VIP genes into these yeast mutants to demonstrate that biochemical complementation takes place. Showing that the plant VIP genes restore InsP7 synthesis in the yeast mutants indicates that the plant VIP genes function in a similar manner as do the yeast VIP genes.

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Health & Medicine ︱ Professor Michel L. Tremblay

Controlling magnesium flux: a central role for the PRL-CNNM complex Magnesium is an essential metal ion for human health.

However, its ability to act as a supplementary therapy against disease is a poorly understood area of science. Professor Michel L. Tremblay and his team at McGill University in Montreal, Canada, are looking to change this though, and are currently investigating the role of the newly discovered pathway, PRL-CNNM, in controlling magnesium’s mechanism of action. This breakthrough discovery has implications for our understanding of cancer, metabolism, circadian rhythm and infectious disease.

M

agnesium (Mg2+) is a metal ion found in the earth, sea and all living creatures, and is essential to life. Present in every single cell of the body, it serves many important functions – especially in supporting biochemical reactions. Notably, it’s an intrinsic component of adenosine triphosphate (Mg-ATP), the universal energy store found in all forms of life.

Its most widely-understood function is its ability to support the physiological processes behind the metabolism of food into energy. Magnesium contributes to the creation of new proteins from amino acids, the contraction and relaxation of muscles, and the regulation of neurotransmitters – chemical substances that transfer messages throughout the brain and nervous system. It also supports gene maintenance, helping to hold proper RNA and DNA structures.

Magnesium is an effective therapeutic agent for a number of health conditions including heart arrhythmia, migraines, asthma and epilepsy. Due to its significant role in sustaining the healthy functioning of organisms, scientists have been investigating its potential role in disease for decades. Research has particularly looked into the processes that regulate magnesium’s presence within the cells.

This is where Professor Michel L. Tremblay comes in. Alongside his team, he has uncovered one of the major pathways that regulates magnesium’s presence within the cells. His team’s

The PRL-CNNM pathway could play a substantial role in controlling the cellular concentration of magnesium

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recent groundbreaking studies suggest that a newly discovered pathway, reliant on interactions between certain protein phosphatase enzymes and magnesium transporters, could play an essential role in controlling the intra-cellular concentration of magnesium which has implications for countless cellular processes and diseases.

PATHWAY TO PROGRESS In recent studies Professor Tremblay shows that the PRL-CNNM pathway could play a role in controlling both the cellular concentration of magnesium, as well as the production of adenosine triphosphate (ATP) – the organic chemical that provides cellular processes with the energy required to drive them.

The “PRL” and “CNNM” acronyms come from the pathway’s reliance on two types of proteins and the way they interact.

The first of these – the Phosphatase of Regenerative Liver (PRL) proteins – come in one of three forms, aptly named PRL1, PRL-2 and PRL-3. These PRL proteins then interact individually with one of four types of magnesium transporter, called CNNM 1, 2, 3 and 4 proteins.

As yet, it is unclear whether these interactions play a role in magnesium transport, or if they simply act as a magnesium sensor for cells.

Nonetheless, results currently show that the more PRL-CNNM protein complexes there are, the higher the level of intracellular magnesium. Think of it like three people (PRL proteins) getting a choice of four cars (CNNM transporters) to drive – driving these cars around causes intracellular magnesium to increase. Indeed, the central role of the PRL-CNNM complex is supported by the fact that it is evolutionarily conserved across all chordates.

MAGNESIUM MUTATIONS Mutating one of the PRL proteins in a study using mice, Professor Tremblay and his colleagues proved this phenomena to be true – establishing a decrease in intracellular magnesium production when PRL-CNNM interactions were inhibited. Through their work, they also found that the PRL enzymes and CNNM proteins worked together and that their collaborative function modulated magnesium levels in cells.

Figure 1 depicts magnesium atoms (yellow balls) that are entering the cells from the action of the two PRL enzymes (green) associated with two CNNMs transporter proteins (pink/purple). This complex is located at the cell membrane.

Once inside the cell, magnesium is recruited by more than 300 proteins (grey) to induce their building block activities in active cells. A second set of molecules that absolutely requires magnesium, are the nucleotides such as ATP (small stick and ball structures) that need an atom of magnesium to be an active source of cellular energy.

PRL-CNNM: A CANCER-CAUSING PATHWAY? The implications of this research are huge.The prominence of PRL gene expression in almost all human cancers, and the fact that PRL levels are higher in metastatic (spreading) tumours compared to non-metastatic tumours, suggests PRL is involved in the process whereby cancers spread.

Professor Tremblay and his team therefore turned their attention towards furthering understanding of this area, investigating the influence of PRL enzymes on cancer growth.

Again using mice as models, their research found that – amazingly – reducing the number of PRL proteins in cancer cells significantly reduced the rate of tumour growth. In other words: lower levels of PRL enzymes, lower levels of cancer growth.

the balance of intracellular magnesium and the pro-oncogenic function of PRL-2 enzymes, pinpointing CNNM complexes as key PRL-binding partners.

of Prof Tremblay and his team’s findings, the role of the PRL-CNNM pathway has now been identified as a major part of cancer’s ability to metastasise.

Interestingly, the team uncovered that, to ensure PRL is sufficiently present in normal cells, a lower magnesium concentration is needed for PRL-2 enzyme expression. Conversely, a higher magnesium concentration

PRL-CNNM: OTHER IMPLICATIONS Although the initial focus of the PRLCNNM complex research was on cancer, the McGill University group have since found it to have other important metabolic functions. They demonstrate that PRL-2 deficient mice have an abnormal physiology, leading to growth retardation, altered body composition and higher mortality rates after birth.

Some of these effects appeared to be sex-dependent – especially in brown adipocytes important for maintaining body temperature – due to PRL being found at higher levels in females than in males. It is possible that female hormones may have a positive effect on PRL gene expression, increasing the amount of available PRL.

The implications of Prof Tremblay’s area of work could be massive, especially within the field of cancer research

Not only that, but the team also found that mutations in PRL or CNNM genes reduce intracellular levels of magnesium and lead to a decrease in the metabolism of cancer cells – preventing them from invading other tissues. This could therefore explain the oncogenic role of the PRL-CNNM pathway, the outcomes of which could be life-changing: should this process occur similarly in humans, different approaches to cancer prevention and therapeutics may be developed.

THE COMPLEX AND THE CANCER So far, Professor Tremblay and his team have found an association between

is needed to inhibit PRL-2 enzyme expression – maintaining the balance of magnesium concentration within the cell. Think of it like balancing a seesaw – you need one person to push up and the other to pull down to create an equilibrium.

However, cancer cells operate differently. These cells rely on a higher magnesium concentration to increase the activity of the PRL-CNNM pathway, ensuring their survival and replication.

It therefore makes sense that PRL expression is often higher in metastatic tumours – instances where cancer spreads to surrounding tissue. Because

PRL-2 deficient mice also showed alterations in their circadian rhythm – the internal clock system that regulates energy expenditure, sleep pattern and metabolism. The importance of this mechanism was recently highlighted

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Mg2+

Mg2+

open

Cell membrane

Transcriptional regulation Protein expression

Structural change Mg2+

CNNMs

Mg2+

Effectors

Active PRL2

24 hr

Mg2+ Mg2+

CNNMs

Inactive

ex. ATP-Mg2+

ex. ATP or

or

Prl2 Cnnms

Tumour Growth

Metabolism

Prl2 Cnnms

closed

24 hr

Figure 2 presents a schematic model of the PRL and CNNM complex cellular function. On the left panel, circadian transcription factors induce the daily synchrony in expression of both PRL and CNNM. When they bind to each other at the cellular surface, they induce a rise in intracellular magnesium. Similarly, PRL and CNNM are expressed differentially in males and females depending on the energy needs of cells. The complex at the membrane promotes the entry of magnesium to support increased metabolic activities in their respective tissues.

On the right panel, once evening arrives, a decrease in the need for magnesium occurs causing a reduction in the expression of both PRL and CNNM. A consequent drop in magnesium concentration and a decrease in metabolism favour resting during the incoming night. In cancer, higher levels of magnesium are needed to sustain the high-energy metabolism of cancer cells. This occurs either through a positive feedback mechanism that is initiated when magnesium is required, thus inducing expression of the PRL enzymes and high magnesium entry or through an increased expression of PRL and CNNM induced by various oncogenes that leads to high magnesium levels, increased cell metabolism and a strengthening of tumour and metastatic burdens.

by the award of the 2017 Nobel Prize in Physiology or Medicine to Jeffrey C. Hall, Michael Rosbash and Michael W. Young for their discoveries of molecular mechanisms controlling the circadian rhythm. Not only that, but another of Prof Tremblay’s studies found evidence suggesting that the PRL-CNNM pathway is involved in energy metabolism – the chemical process animals and humans use to turn ingested food into energy.

Previous research had already pinpointed magnesium as a regulator of metabolic and circadian processes, but in a recent report Prof Tremblay identified that the PRL-CNNM proteins were directly regulated by proteins responsible for controlling circadian rhythm. During the morning, higher levels of PRL-CNNM complexes are present, which increases the concentration of intracellular magnesium. This facilitates metabolic output, awakening us and providing us with an active start to the day. In

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contrast, at the end of the day, the intracellular magnesium concentration depletes, allowing our “cellular furnace“ to cool down and encouraging us to rest.

Lastly, but rather excitingly, the PRLCNNM complex has been seen to be modulated by virus infection. Findings from Profs Tremblay and Teodoro’s McGill laboratories have suggested that – in a similar way to cancer – certain viruses rely on activating the PRL-CNNM complex, to produce the higher levels of magnesium required for optimal viral reproduction. This also seems to be true of many human parasites, including the malarial agent, which in collaboration with Dr Tewari (University of Nottingham) suggests a new possibility for fighting these pathogens.

THE PATHWAY FORWARD Prof Tremblay and his team’s research provides vital evidence about the newly-discovered PRL-CNNM protein

complex, and the biological importance of controlling intracellular magnesium concentrations. Their research will inform further studies looking into the role of magnesium, both metabolically and therapeutically – within disease prevention.

The implications for this area of work are huge, especially within the field of cancer research. Tumour growth is associated with the PRL-CNNM pathway and the flux control of intracellular magnesium. Improving understanding of this area, and extending research insight from mice to humans, will not only broaden our knowledge of the molecular processes involved in controlling magnesium concentration, but it could also lead to novel therapies both in oncology, and against infectious disease.

One thing is for sure: magnesium’s role has never looked so important.

Behind the Bench Professor Michel L Tremblay

E: Michel.tremblay@mcgill.ca T: +1 514 398 7290/8480 W: https://mcgillgcrc.com/research/members/tremblay

Research Objectives Professor Tremblay’s research focuses on investigating Protein Tyrosine Phosphatases (PTPases), specifically looking into their roles in a variety of cellular processes such as cell growth, differentiation, and cancer.

Funding Canadian Institutes of Health Research (CIHR)

• Prof Alphonso Martinez de la Cruz (Center of Excellence Severo Ochoa, Bilbao) • Prof Andreas Bikfalvi (University de Bordeaux) • Professor Rita Tewari (University of Nottingham) • Professor Daniella Bucella (New-York University) • Professor Joost Hoenderop (Rabdoub University)

Collaborators • Dr Noriko Uetani (McGill University) • Dr Serge Hardy (McGill University) • Prof Jose Teodoro (McGill University)

Bio Michel L Tremblay is a Professor, a James McGill Professor and holder of the Jeanne and Jean-Louis Lévesque

Q&A

When and how did you first start being interested in the role of magnesium for the functioning and potential malfunctioning of the human body?

When we first identified that mice engineered to lack functional PRL genes had changes in their blood magnesium concentration, and that PRL proteins were binding to the magnesium concentration regulator CNNMs proteins.

What would you say were your most noteworthy findings so far and how might these be applied to human studies in future?

• Cancer cells must increase their magnesium intracellular levels to maintain a high metabolic rate through the PRL-CNNM complex.

• Many infectious agents may target this complex – influencing cell metabolism – to favour their infectious cycle and replication.

• Magnesium controlling the beginning

of the daily circadian rhythm does this in part by regulating this complex.

• Females and males have differences in their levels of the complex, thus affecting the metabolic output and mechanism of using glucose differently in males and females.

• The control of magnesium levels by the complex likely explains the multiple sites in cells affected by PRL. This is because over 300 enzymes are known to depend on magnesium to maintain their activities.

In the mouse studies you conducted, what evidence suggests that pathways controlling the magnesium flux could be involved in diseases and what further research needs to be carried out in order to ascertain that?

The most solid findings come from our research showing that, by removing PRL in tumour cells, cancer cells were less invasive and oncogenic in the animal model. When we examined these cancer cells with PRL reductions, they had less intra-cellular magnesium and were less oncogenic.

Chair in Cancer Research, at McGill University. A graduate of the Université de Sherbrooke (MSc) and of McMaster University (PhD), he completed his post-doctoral training at the National Institutes of Health in Bethesda.

Contact Professor Michel L Tremblay Department of Biochemistry Rosalind and Morris Goodman Cancer Research Centre Cancer Research Building 1160 Pine Avenue West Office: Room 617; Lab: Room 603 Montreal, Quebec H3A 1A3, Canada

You recently met with other international scientists who are also carrying out research in this area, what were some of the most prominent points you discussed?

The structure of the complex – the existence of mutations in the CNNM human population and their association with kidney hypomagnesonemia diseases. We also discussed the role of the complex in modulating infectious agents, as well as how PRL-CNNM modulates magnesium concentration, and how the PRL-CNNM genes and proteins are regulated.

What are your next steps in terms of future investigation?

Developing novel inhibitor molecules and identifying infectious agent proteins that interact with this complex.

We also hope to identify how the complex controls energy expenditure and other cellular functions.

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Health & Medicine ︹ Professor Raymond Harris and Professor Ming-Zhi Zhang

Blood, skin and bone: the complex control of blood pressure

It is well-known that excessive salt intake can be a risk factor for high blood pressure. But how this effect is mediated – and why some people are more susceptible than others – remains up for debate. Collaborators Professor Raymond Harris and Professor Ming-Zhi Zhang, at Vanderbilt University School of Medicine, have uncovered a novel role for immune cells derived from bone marrow in salt-sensitive high blood pressure, hinting at potential new ways to manage and treat the condition.

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High-salt diet

Skin

Kidney Macrophage

VEGF-C

?

?

Arachidonic acid

T cells Cytokines NCC

COX-2

Lymphangiogenesis

mPGES-1 Na+ content

EP4R Na+ content

PGE2

The COX-2/mPGES-1/EP4 receptor pathway in macrophages maintains sodium homeostasis by influencing salt secretion in the kidney and extrarenal sodium storage.

H

igh blood pressure (hypertension) may affect as many as a quarter of the developed world population, and is a significant risk factor for cardiovascular conditions such as heart attacks and strokes. Although the causes of high blood pressure are many and complex, most cases are at least partially triggered by a high salt diet: high levels of sodium pull water into the blood stream, increasing the pressure as it passes through the blood vessels. Therefore, the relationship between dietary salt and hypertension is an important public health concern that needs addressing.

Inhibition of the cyclooxygenaseprostaglandin pathway has already been linked experimentally to clinically significant salt-sensitive hypertension.

Until recently, the action of the cyclooxygenase-prostaglandin pathway in maintaining stable blood pressure was thought to be limited to the kidneys – the organs that, after all, play the major role

on the bone marrow, the source of ‘haematopoietic’ cells: stem cells responsible for making all other types of blood cells, including the white blood cells of the immune system.

HYPERTENSION THROUGH TRANSPLANTATION Professors Harris and Zhang took laboratory mice with a normal, functioning cyclooxygenaseprostaglandin pathway, which could maintain normal blood pressure regardless of the amount of salt ingested. They then transplanted bone marrow cells into these mice, in which the gene for COX-2 had been switched off. When the mice were then fed a high salt diet, their blood pressure increased.

This showed that, without COX-2 in their bone marrow – and therefore also in the blood cells derived from it – the mice were less able to deal with the excess salt, despite having functional COX-2 molecules in other parts of their body including the kidneys.

Previous paradigms about the development of salt-sensitive hypertension are incomplete

A HEALTHY BALANCE When our bodies function correctly, they can counteract excess salt consumption and maintain ‘homeostasis’ – a constant balance of the chemicals, hormones, and other parameters within the body.

A key mediator of salt homeostasis is the cyclooxygenase-prostaglandin pathway, which synthesises hormone-like signalling molecules (prostaglandins) responsible for a range of physiological effects – including inflammation, dilation of blood vessels, and the production of blood-pressure-related hormones.

in maintaining our bodies’ salt and water balance. High salt diets were known to increase production of the pathway’s key, rate-limiting enzyme, cyclooxygenase-2 (COX-2) in both kidney tissue and white blood cells circulating in the kidney, resulting in an excretion of excess sodium.

The work of Professor Harris and others, however, is beginning to implicate a range of other tissues in salt homeostasis, including the immune system and even the skin. In particular, Harris and Zhang’s research focuses

Professors Harris and Zhang concluded that cells produced in the bone marrow must play a hitherto-overlooked role in maintaining salt balance and blood

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pressure through the cyclooxygenaseprostaglandin pathway. Not only that, but as Prof Harris points out: “Previous paradigms about the development of salt-sensitive hypertension are incomplete.” When the experiment was reversed, so that mice lacking in COX-2 throughout their body were transplanted with normal bone marrow, their hypertension improved, confirming the new findings.

Further investigations of the transplanted mice found changes in the population of at least two forms of white blood cells, macrophages (which usually engulf invading pathogens and debris) and T cells (which recognise and attack pathogens through specific antigens, contributing to immunity), in their kidneys and skin. Prof Harris suggests that prostaglandins generated from bone marrow-derived white blood cells may partner with those from the kidney to prevent hypertension caused by a high salt diet.

Very similar results were found when mice were transplanted with macrophages lacking a key receptor further on in the cyclooxygenaseprostaglandin pathway, EP4, which mediates prostaglandin signalling, suggesting that disrupting either prostaglandin production (by COX2) or reception (by EP4) in these cells can cause salt-sensitive hypertension.

Overall, the study suggests an unexpectedly important role for white blood cells in salt homeostasis, generated in the bone marrow but acting at remote sites including the kidneys and skin. However, according to Harris and Zhang, the underlying mechanisms appear to be complex and multifactorial.

STOP POPPING THE PAINKILLERS The cyclooxygenase-prostaglandin pathway has many different roles in the human body besides salt homeostasis, for instance mediating inflammation and pain. Some of the world’s most popular painkillers – the non-steroidal anti-inflammatory drugs (NSAIDs), such as aspirin and ibuprofen – act by blocking the action of COX-2 to prevent inflammation triggered by prostaglandins. Thus, side-effects of regular NSAID use include both salt-

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Cells produced in the bone marrow must play a hitherto-overlooked role in maintaining salt balance and blood pressure sensitive hypertension and peripheral oedema, an associated swelling of the extremities caused by a build-up of fluid in the spaces between tissues of the body. Harris and Zhang’s work shows that these effects may be more complex and widespread than previously thought.

Their results so far are tantalising, suggesting a completely novel route to hypertension in mammals. In a project funded by the US National Institutes of Health, they now propose to further tease out the complex role of the prostaglandins produced by COX-2 in cells derived in the bone marrow.

They want to find out exactly how the pathway is triggered into action by the presence of excess salt, how it relates to

the occurrence of peripheral oedema, and to quantify the relative roles of macrophages in the different organs of the body in managing hypertension.

In light of their findings so far, Harris and Zhang propose to test whether hypertension linked specifically to NSAID use occurs through the inhibition of the cyclooxygenase-prostaglandin pathway in bone-marrow derived cells.

They are also considering the possibility of activating the EP4 receptor through drugs, as a new treatment for saltsensitive hypertension. Their results may ultimately suggest ways to both treat and prevent this widespread, chronic medical condition.

Behind the Bench Dr Raymond C Harris

Ming-Zhi Zhang

E: raymond.harris@Vanderbilt.Edu T: +1 615 322 2150 W: http://medicine.mc.vanderbilt.edu/nephrol_RaymondHarris

Research Objectives Professors Harris and Zhang’s work specifically focuses on understanding the role of COX-2 and prostaglandins in the kidney, studying the underlying mechanisms of diabetic nephropathy and determining mechanisms of growth and repair in response to acute and chronic injury.

Funding National Institutes of Health (NIH) National Institute of Diabetes Digestive and Kidney Diseases (NIDDK) Bio Raymond Harris, MD is currently the Ann and Roscoe R Robinson

Q&A

How does a high salt diet lead to hypertension?

Although there continues to be some controversy about the role of dietary science, most epidemiologic evidence indicates an association with hypertension. A significant percentage of the population is “salt sensitive”, and increased salt ingestion leads to either development or exacerbation of hypertension.

How do the different organs now known to be involved in salt homeostasis (for instance, kidneys, bone marrow and skin) communicate with one another?

Our understanding of salt homeostasis is rapidly changing. Although new studies now clearly show that salt can accumulate in both skin and muscle,

Professor of Medicine and Director of the Vanderbilt Center for Kidney Diseases. Research in the Harris lab has focused on understanding mechanisms underlying the pathogenesis of both acute and chronic kidney disease.

Ming-Zhi Zhang graduated from XuZhou Medical University in China and is an Associate Professor of Medicine and Cancer Biology in Division of Nephrology at Vanderbilt. His research focuses on cyclooxygenase-2/prostaglandin signalling pathway in macrophage polarisation and its role in regulation of blood pressure homeostasis.

how these organs communicate with the kidney, which is ultimately responsible for salt and water homeostasis, remains uncertain. However, there is increasing evidence that increased salt ingestion activates an inflammatory response in the affected organs, which may be a major predisposing factor for development of salt-sensitive hypertension.

How can you be sure that your results from mouse studies are applicable to humans?

Ongoing clinical studies using sodium MRI clearly show that humans also

Contact Raymond C Harris MD Ann and Roscoe Robinson Professor of Medicine Chief, Division of Nephrology and Hypertension C-3121 MCN Vanderbilt University School of Medicine Nashville, Tennessee 37232 USA

accumulate sodium in skin and muscle, and studies with COX-2 selective inhibitors have shown a predisposition for development or exacerbation of salt-sensitive hypertension.

What do your results mean for the use of NSAIDs? It is well known that both non-selective and COX-2 selective NSAIDs can predispose to development of saltsensitive hypertension. These studies provide a mechanistic understanding of underlying mechanisms.

It is well known that both non-selective and COX-2 selective NSAIDs can predispose to development of saltsensitive hypertension

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Thought Leader

Canadian Blood Services: The road to regaining public trust

The need for blood is constant; so is the need for donations. Every day, all the hospitals and clinics in Canada need blood and blood products to treat patients, since most surgical interventions and a great number of medical procedures require blood transfusions. This is where Canadian Blood Services comes in.

Canadian Blood Services is a non-profit charitable organisation with a mission to manage the bloody supply for Canadians and provide a safe, secure, costeffective and accessible supply of quality blood, blood products and their alternatives. We spoke with Canadian Blood Services’ CEO Dr Graham Sher at Research Outreach, to discuss this and more, in greater detail.

I

n the 1980s, more than 2,000 people in Canada were infected with HIV and over 30,000 with hepatitis C after they had been administered tainted blood products. In the wake of disaster, an inquiry led by Justice Horace Krever exposed years of negligence, bureaucratic inertia and at times corruption at the Canadian Red Cross Society, then in charge of the blood donation system. In consequence of

Can you tell us what attracted you to Canadian Blood Services and what your role there involves?

When I was asked by the newly founded Canadian Blood Services to join the organisation as a vice-president of medical, scientific and clinical management back in 1998, I worked as a physician and scientist on staff at the Toronto Hospital and on faculty at the University of Toronto. I had no plans to

Patients depend on us to manage a safe, secure and cost-effective blood system Krever’s recommendations, 1998 saw the foundation of Canadian Blood Services that replaced Canadian Red Cross Society in managing national blood supplies. It took nearly 20 years of Canadian Blood Services’ leadership and dedication to rebuild the Canadian blood system, make it an international success story, and regain the public trust.

We recently caught up with Dr Sher at Research Outreach and talked with him about the organisation’s role, his role as CEO over the last 20 years and the future of blood donation in Canada.

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leave my research lab or teaching role, but the opportunity to move beyond the individual patient level and have a greater impact on the wider healthcare system, and ultimately, serve more patients, was too appealing to pass up.

A few years later, in 2001, I became a CEO. Since then, I have been leading the organisation through a multiyear transformation journey aimed at redesigning the entire service delivery model, introducing best business practices, and growing a culture of high performance.

As part of transforming the national blood system, I have also led Canadian Blood Services through a significant expansion in its scope of services, which led to the organisation assuming a national leadership and coordinating role for both organ and tissue donation and transplantation in Canada, and the development of Canada’s national cord blood banking programme. Finally, I co-founded and continue to actively participate in an international alliance

of national blood system operators, with the focus of benchmarking, best practice sharing and global policy advancement in our sector.

Can you give us an overview of what Canadian Blood Services does and what its aims are?

Canadian Blood Services manages the national supply of blood, blood products, stem cells and related services for all provinces and territories

(except Quebec). We operate within the larger health-care system of transfusion and transplantation medicine in Canada. Patients depend on us to manage a safe, secure, and cost-effective blood system.

The organisation collects, tests and manufactures blood, blood products and stem cells, and plays an integral role in organ and tissue donation and transplantation.

Our responsibilities also include: running national patient registries for organ donation and transplantation; operating the OneMatch Stem Cell and Marrow Network, which matches donors to patients that require stem cells transplants; as well as Canadian Blood Services’ Cord Blood Bank.

Moreover, Canadian Blood Services is involved in research and development efforts focused on several areas of

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transfusion and transplantation science and medicine. We draw in experts from various disciplines who together can bring innovative thinking to bear on real problems.

Why was the decision made to centralise Canada’s blood services in 1998?

What have been the organisation’s key achievements in that period?

This decision was made in the wake of the tainted blood tragedy, the largest public health catastrophe in the country’s history. Justice Horace Krever led an inquiry into the scandal and published his report in 1997. Following one of his recommendations, Canadian Blood Services was created and trusted with a mission to ensure such a disaster would never happen again.

Back in 1998, we inherited a fragmented blood supply system plagued with critical quality failures, badly ageing facilities, and structural complexity.

When I became CEO, I recognised that to transform the system in a long-term and sustainable manner we needed to move from crisis management to strategic management. I set about creating a business framework that allowed us to plan for changes over a long horizon and to move the organisation to a much higher level of operational stability and performance success. Since then, the organisation has integrated about a dozen regional, disconnected supply chains into one seamless national system.

Today, whether patients are in Victoria, Iqaluit, St. John’s, or anywhere in between, they can count on the same high-quality product when they need it, without geographical or financial barriers. When new pathogens emerge, like West Nile virus, SARS, H1N1, or Zika, Canadian Blood Services is at the forefront of an international community of scientists working together to protect patients at home and around the globe.

Can you describe Canadian Blood Services’ role in the national formulary of plasma-derived medicine, and synthetic alternatives? What are the benefits and disadvantages of this system?

As the steward of the public blood

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Dr Graham Sher, CEO of Canadian Blood Services since 2001

system, Canadian Blood Services is the trusted supplier of plasma and plasma protein products for patients in Canada.

We manage a pan-Canadian formulary of approximately 45 brands of plasma protein products, which we bulkpurchase on behalf of provincial governments. For the patient’s standpoint, our product selection process supports patients’ and physicians’ involvement in decisionmaking. It’s also cost-effective. Bulkbuying and price negotiations bring significant savings.

In addition to collecting plasma for transfusion, we collect plasma to be used as a raw material to produce immune globulin (Ig), a critical,

lifesaving drug in very high demand.

It is our responsibility to ensure enough plasma goes to manufacturing Ig for Canadian patients.

We currently only collect enough plasma to meet about 17% of the demand for Ig. To meet patient needs, we purchase the remainder of the necessary product from the commercial plasma industry. We plan to expand plasma collections in Canada to ensure a secure supply of plasma for Ig for Canadian patients.

Can you tell us more about the OneMatch Stem Cell and Marrow Network?

Fewer than 25% of patients who need stem cell transplants find a compatible

Thought Leader donor in their own family. The rest rely on those who have volunteered to donate stem cells to anyone in need.

Thanks to the OneMatch programme we can now search more than 23 million donors in more than 70 registries in other countries when we need to find a match. By making donor data available worldwide, international registries have significantly increased the odds of finding a matching donor for any patient anywhere in the world.

What role does Canadian Blood Services have in improving the national levels of blood and organ donation?

Improving the national inventory of blood is ongoing. We help

With more than 800 transplants resulting from the KPD and HSP programmes combined, many Canadians have received transplants that may never have otherwise occurred.

You are a haematologist by training, and an expert in transfusion medicine.

How did you first become interested in this field?

In short, while doing my undergraduate medical training, I was interested in neurology, and therefore, destined to become a neurologist. I even did a PhD degree (simultaneous with my medical degree) in neuroscience. In my secondto-last year of medical training, while rotating through a general medical unit,

to haematology rather than neurology.

And, as they say, the rest is history!

What challenges are likely to face blood and organ service provision in Canada during the next decade?

We need to increase the amount of plasma we collect to ensure a secure supply of plasma needed to manufacture immune globulin (Ig), a critical lifesaving drug, for Canadian patients. We plan to do this within our current voluntary, unpaid system.

We also need to recruit more blood and organ donors. We are focused on connecting with an ever-changing population of donors; the population in Canada is shifting to metropolitan

We draw in experts from various disciplines who together can bring innovative thinking to bear on real problems hospitals improve blood utilisation and surveillance and have found that educating consumers, donors, physicians and other health professionals is key to managing utilisation of blood and blood products.

Our work in organ donation and transplantation may be less well known.

Let me name a few of our initiatives aimed at improving matters in this field. Through the Kidney Paired Donation (KPD) programme, we facilitate medically compatible kidney transplants through chains of donor exchanges from medically incompatible pairs. The Highly Sensitized Patient Kidney (HSP) programme improves chances of a kidney transplant for hard-to-match patients. The National Organ Waitlist (NOW) is a real-time data source for non-renal patients throughout Canada.

We work with stakeholders, partners and physician groups to evolve knowledge, policy and technology.

This leads to increased donation and transplantation rates, gives patients the best possible chances to receive transplants with optimal outcomes, and gives families the opportunity to honour their loved one’s wishes to become an organ donor.

I become the responsible physician looking after a young patient with acute myeloid leukaemia. We managed to get the patient into remission, and ready her for a lifesaving bone marrow transplant from her only sibling, an older brother.

Back then, unrelated bone marrow stem cell transplants had not become standard of care, and so it was a related transplant or nothing. Fortunately, this patient’s brother was a perfect “6 out of 6” match for her, so all was hopeful and positive. Tragically, the day before the planned bone marrow collection, her brother (and donor) was killed in a motor vehicle accident. As such, no transplant option existed for this patient, and she died some months later after relapse of her leukaemia.

These two — she 16, he 19 — were the only children of two remarkable parents, who themselves had been children survivors of the Holocaust in Europe. The capacity of these parents to deal with the enormity of the grief and loss they experienced was the most remarkable display of humanity I had ever witnessed. It was profoundly humbling and entirely remarkable, in ways that defy description. Their impact on me personally was so significant, that I switched paths and dedicated my future career

areas, and we need to go where the people go to operate as efficiently as possible and make blood donation as convenient as possible for donors.

• For more information on the Canadian Blood Services, their ground-breaking research and blood donation, please visit their website at blood.ca/en.

Contact Dr Graham Sher Canadian Blood Services 1800 Alta Vista Drive Ottawa Ontario K1G 4J5 Canada E: feedback@blood.ca T: +1 613 739 2300 F: +1 613 731 1411 W: blood.ca/en

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Health & Medicine ︱ Professor André Carpentier

Disordered fat storage enhances development of type 2 diabetes Diabetes is a devastating, lifechanging condition. Professor André Carpentier and his colleagues from Université de Sherbrooke are exploring how obesity increases the risk of type 2 diabetes. After all, abnormal fat storage can lead to insulin resistance, which endangers organ function. Through his and his team’s research, Dr Carpentier aims to develop therapeutic measures which reverse detrimental pre-diabetic effects.

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n 2015, diabetes mellitus was the sixth leading cause of death worldwide.

This chronic metabolic disorder results in insufficient glucose uptake from the blood, due to abnormal insulin activity.

Pancreatic cells secrete the hormone insulin into the blood where it triggers skeletal muscle, liver and fat cells to absorb and store glucose. Irregular insulin activity can lead to hyperglycaemia (high blood glucose levels) which in the long-term can cause kidney disease, cardiovascular complications, foot ulcers and eye damage.

Insufficient insulin activity arises from either: i) an autoimmune response, leading to a loss of pancreatic cells and dangerously low insulin levels (type 1 diabetes), or ii) cells that become insulin resistant and are unable to respond to insulin signals with an inability of pancreatic cells to compensate by

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secreting enough insulin to counter this insulin resistance (type 2 diabetes [T2D]).

Environmental factors such as obesity, smoking or a high, low-quality fat and carbohydrate diet all increase the risk of T2D development.

Although preventable, T2D is much more common, accounting for 90% of all diabetes cases worldwide – a statistic that is increasing rapidly. Globally, T2D cases have quadrupled since 1980 and currently 8% of the global population suffer from it. Increasing incidence rates are also costly, with direct medical costs for T2D exceeding $825 billion globally.

Clearly, there is an urgent need for therapeutic options to prevent T2D progression at the pre-diabetes stage, where blood glucose levels are high, but do not cross the ‘diabetic’ threshold. Professor Carpentier and

Above: A healthy individual undergoing a study to measure brown adipose tissue (BAT) metabolic response to acute cold exposure using positron emission tomography coupled to computed tomography (PET/CT), indirect calorimetry and various metabolic tracers administered intravenously

his team are therefore researching how obesity-related abnormal fat storage/ metabolism drives insulin resistance.

Through this research, the team have been able to explore innovative measures to prevent T2D.

lower in obese individuals, compared to healthy individuals. Consequently, lean tissues such as the heart, liver and skeletal muscles are overexposed to fatty acids, which leads to lipotoxicity, organ dysfunction and insulin resistance.

DIET-RELATED INSULIN RESISTANCE Studies have shown that there is a strong link between a diet high in saturated fat and insulin resistance. Although the exact mechanisms are unknown, evidence suggests that the disordered storage of dietary fatty acids (DFA) in white adipose tissue (WAT) plays a key role. WAT is an extremely dynamic energy store and overnutrition can result in adipose tissue remodelling.

Adipocytes can then undergo drastic alterations in number, size and metabolic activity. Despite adipose tissue expansion, DFA storage efficiency per mass of adipose tissue is significantly

PET IMAGING To investigate altered fat distribution, Professor Carpentier used an innovative molecular imaging tool to quantify organ-specific DFA partitioning.

Previously, isotropic tracers have been used to assess organ-specific

be applied to all skeletal muscle/ adipose tissues simultaneously, limiting the measurement of whole body DFA partitioning, and secondly, isotopic tracers are invasive, making it challenging to study internal organs such as the heart.

However, Professor Carpentier’s method of using non-invasive PET (Positron Emission Tomography) imaging overcomes these limitations.

PET is a sensitive imaging technique which can detect tiny (picomolar) tracer concentrations. Professor Carpentier performed a study in which healthy subjects were orally administered the radioactively labelled tracer FTHA (14-R,S-F-fluoro-6-thia-heptadecanoic acid), a long-chain fatty acid analogue.

Sequential, wholebody PET acquisition was then performed over six hours.

During this period, all tracer movements were recorded as a three-dimensional image to highlight whole-body tracer partitioning.

Essentially, this allowed DFA fat storage in different tissues and organs to be measured. Results showed that in healthy

Inspired by their promising results, Professor Carpentier and his colleagues are now exploring other mechanisms to prevent development of type 2 diabetes DFA uptake. Radioactively labelling DFA enables detectors to track movement and deposition location.

However, isotropic tracers have several limitations: firstly, this method cannot

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An axial view of the heart using PET/CT after intravenous injection of 11C-palmitate, a free fatty acid tracer. Incremental tracer uptake is depicted using a blue to red color scheme (low to high uptake)

men, DFA storage was high in the liver and heart. However, skeletal muscles and subcutaneous adipose tissue storage contained relatively low levels of DFA per volume of tissue.

Professor Carpentier also investigated DFA partitioning in brown adipose tissue (BAT). The function of BAT is thermoregulation via thermogenesis – heat generation by oxidation of fat stores to raise body temperature. Subjects were administered oral FTHA and exposed to low temperatures of 18°C. PET imaging results confirmed that BAT does uptake DFA during cold exposure.

However, BAT DFA partitioning per volume of tissue was around 83% lower than the liver and 55% lower than in the heart, contributing to only 0.3% of total body DFA clearance.

DISORDERED FAT STORAGE The team also explored organ-specific DFA partitioning per volume of tissue in pre-diabetic and obese (with normal glucose levels) subjects. Interestingly, results support the notion that a prediabetic state results in ineffective adipose DFA tissue storage, which is strongly related to obesity. In terms of lean organ partitioning, DFA storage did not differ between healthy, obese

and pre-diabetic individuals for the liver or skeletal muscle. However, Professor Carpentier found that cardiac DFA uptake is significantly greater in prediabetic individuals. In both healthy and obese individuals, 2–3% of DFAs are stored in the heart. However, this rises to 4% in pre-diabetic individuals, which can lead to severe cardiac complications, such as left ventricular dysfunction, and potential heart failure. Although this finding is alarming, Professor Carpentier showed that by making lifestyle changes leading to a modest reduction of body fat, this can reverse DFA channelling to the heart. Inspired by this promising result, Professor Carpentier and his colleagues are now exploring other mechanisms to prevent T2D from developing.

BARIATRIC SURGERY TO TREAT T2D Bariatric surgery is where the stomach is reduced in size by stapling and the small intestine bypassed, limiting food intake and intestinal fat absorption.

The team investigated the benefits of bariatric surgery in terms of improved WAT activity and reduced overexposure of lean tissue to DFA in obese individuals with and without T2D. Twelve months following bariatric surgery, on average,

Professor Carpentier and his team have greatly advanced our knowledge of how abnormal fat distribution enhances the risk of type 2 diabetes development

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Coronal view of PET/CT acquisition after intravenous injection of 18Fluorodeoxyglucose, a glucose tracer, during acute cold exposure.

The bright yellow areas depict regions with high glucose uptake, including the brain, supraclavicular brown adipose tissues, the heart, and the liver

an excess weight loss of 85% was achieved. Furthermore, all T2D subjects were in partial or complete remission after three months following surgery.

This is due to improved insulin sensitivity in accordance with increased weight loss, better fatty acid handling by lean tissues, and significant reduction of WAT cell size.

FUTURE RESEARCH Overall, Professor Carpentier and his team have greatly contributed to advancing our knowledge of how abnormal fat distribution enhances the risk of T2D development and how preventative mechanisms such as bariatric surgery can reverse these detrimental effects. However, several questions remain unanswered. For example, why is DFA channelling to the heart increased in pre-diabetic individuals and whether it could be targeted to reduce the risk of heart failure, and how exactly does lean tissue lipotoxicity result in insulin resistance?

Behind the Bench Professeur André C Carpentier

E: Andre.Carpentier@USherbrooke.ca T: +1 819 564 5241 W: www.usherbrooke.ca/dep-medecine/ recherche/professeurs-ayant-des-activites-de-recherche/endocrinologie/pr-andre-carpentier/ Research Objectives Professor Carpentier’s research interests include: 1) the role of postprandial fatty acid metabolism in the development of type 2 diabetes and cardiovascular diseases; 2) the investigation of brown adipose tissue metabolism in diabetes; and 3) the antidiabetic mechanisms of bariatric surgery.

Funding • Canadian Institutes of Health Research (CIHR) • Canadian Diabetes Association

Q&A

How does excess fat cause insulin resistance?

Many mechanisms have been described including: 1) intracellular accumulation of reactive fat metabolites such as diacylglycerols and ceramides; 2) mitochondrial fatty acid overload with increased production of reactive oxygen species (ROS); 3) endoplasmic reticulum stress that also may lead to exaggerated ROS production; 4) changes in phosphorylation of the insulin signalling pathways by activation of cellular inflammation; and 5) activation of Toll-like receptors, triggering cellular inflammation pathways. In general, saturated fats have more profound effects on these pathways than unsaturated fats.

What cardiac complications could result from increased pre-diabetic channelling of fatty acids to the heart?

We and others have observed higher cardiac oxygen consumption, but reduced glucose utilisation when fat utilisation by the heart was increased.

This may lead to enhanced susceptibility of the heart to lack oxygen when its blood flow is hampered. It may also lead to intracellular acidification, leading in turn to chronic heart damage. In our studies, whenever cardiac DFA uptake was enhanced, we found reduced cardiac pumping of the blood.

Collaborators • Éric Turcotte, MD, Université de Sherbrooke • Brigitte Guérin, PhD, Uni. de Sherbrooke • Martin Lepage, PhD, Uni. de Sherbrooke • Roger Lecomte, PhD, Uni. de Sherbrooke • Denis Richard, PhD, Université Laval • André Tchernof, PhD, Université Laval • François Haman, PhD, University of Ottawa Bio Professor Carpentier is the recipient of the GSK Research Chair in Diabetes of Université de Sherbrooke and professor, endocrinologist-lipidologist and clinician Why is PET imaging advantageous to observe abnormal fat metabolism?

PET is the most sensitive imaging modality to non-invasively detect labels administered into the body. With proper setup and expertise, one can design almost any labelled molecule that can then be detected by PET in any organ of the body.

Because PET is so sensitive, you can safely administer very small amounts of these labels that are chemically altered to suit the needs of the investigations pursued. With PET, you also are not limited to one organ or tissue, but you can simultaneously study the uptake and metabolism of your preferred PET label in all tissues in the field of view of your scanner. That makes PET an ideal tool for multi-organ integrative metabolic studies.

Could diet-induced thermogenesis help to prevent type 2 diabetes?

This is an important question to which we still do not have a definitive answer.

In animal experiments, there is a strong body of evidence that brown adipose tissues (BAT) may strongly increase energy expenditure and reduce the metabolic abnormalities leading to type 2 diabetes. In rodents, there is also strong evidence that BAT is an important effector of diet-induced thermogenesis. In humans, we now know that BAT can utilise DFA and, therefore, may somewhat contribute to energy expenditure occurring after meals. We however found that, at least during cold exposure, the relative contribution of BAT is small. There are many limitations to current work,

scientist in the Departments of Medicine, Faculty of Medicine at the Université de Sherbrooke. He is also the director of the university’s Centre de recherche sur le diabète, l’obésité et les complications cardiovasculaires.

Contact Professeur André C Carpentier MD FRCPC Chaire GSK sur le Diabète de l’Université de Sherbrooke/GSK Chair in Diabetes of Université de Sherbrooke Département de médecine Centre de recherche du CHUS Université de Sherbrooke Canada however, including the fact that no one, including our group, is able to precisely quantify the total volume of BAT in the body. Therefore, the precise contribution of BAT thermogenesis may at the moment be severely underestimated, especially in individuals suffering from obesity and type 2 diabetes. This is the next outstanding question that needs to be addressed in the field of human BAT research.

What are your research goals over the next five years?

In addition to quantifying more accurately BAT thermogenesis in individuals with obesity and type 2 diabetes, our major goals will be to establish the mechanisms of enhanced cardiac DFA uptake in men and women with pre-diabetes and whether this functional biomarker can be used to successfully predict and monitor the benefits of nutritional, pharmacological and surgical interventions to treat type 2 diabetes.

The quantification of BAT thermogenesis and DFA organ-specific partitioning may serve as functional biomarkers leading to more individualised preventive and therapeutic approaches of people at risk of suffering from type 2 diabetes and its complications.

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Health & Medicine ︱ Dr Lixing Yang

The genomics of cancer Many cancers are associated with changes to our genetic material, DNA. These may be small, single unit substitutions, large rearrangements such as deletions or duplications of a part of the DNA sequence, or various other forms of mutations. Although the smaller substitutions have been more intensively studied, Dr Lixing Yang, of the University of Chicago, focuses on uncovering changes at the larger end of the spectrum. His work suggests certain key variations may be connected to particular forms of the disease.

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W

hen we hear the term ‘genetic disease’, we probably think of something like cystic fibrosis, or Huntington’s Disease. But, increasingly, cancers are also being linked to genetic causes, whether an inherited trait found in every cell of the body, or a change occurring in just the cells of a tumour itself.

REARRANGING THE GENOME Genes are held on chromosomes, aggregates of DNA and protein found in almost every cell of the body. DNA is the blueprint of most living organisms.

Our DNA is made up of a sequence of paired sub-units known as nucleotides or bases, denoted by the letters A, C, G and T. DNA changes can be as

small as a single substitution: a T for an A, say, or a C for a G; or they can be as large as an entire portion of one chromosome – thousands or millions of As, Cs, Gs and Ts – being transferred to another. The larger changes may affect an order of magnitude more DNA than single nucleotide variations, yet the functional impact of such ‘genomic rearrangements’ has until now been largely unexplored.

Genomic (or ‘structural’) rearrangements are commonly generated during DNA replication, or when the cellular machinery that repairs damaged DNA is not functioning correctly. They may take many forms and arise from multiple different events, such as deletions,

duplications, insertions, inversions or translocations of DNA within and between chromosomes.

A DNA double helix

Recent advances in technology now make it cheaper, quicker and easier than ever before to determine the entire sequence of the DNA held in the cells of an organism. This ‘whole genome sequencing’ has revolutionised many areas of biology – and cancer biology is no exception. Dr Yang’s research focuses on using whole genome data to discover larger scale structural rearrangements of our genetic material that may cause, or contribute to, the development of cancer, and to determine the relative contributions of these different kinds of rearrangements in different kinds of cancers.

MEERKAT TO THE RESCUE The human genome is over three billion base pairs long – so searching for genomic rearrangements in its sequence is no easy matter. For this reason, Dr Yang has developed a novel computer program to facilitate his quest. The algorithm, Meerkat, can both identify more complex rearrangements than available methods and ensure high levels of accuracy in pinpointing the location and nature of structural rearrangements of the genome. Dr Yang has already used Meerkat to catalogue structural variations found in over a thousand tumours from patients with dozens of different types of cancer.

His research has shown that in tumours, two forms of DNA damage – breaks in both strands of the DNA double helix and errors in DNA replication – drive the majority of events leading to genomic rearrangements. For deletiontype structural changes, for instance, around 20% result from complex errors occurring when DNA is replicated.

Focusing on particular types of cancer, Dr Yang has found that in patients with a rare form of kidney cancer, chromophobe renal cell carcinoma, one gene, known as TERT, is sometimes activated. TERT is involved in cell proliferation and is essential to keep the chromosomes stable after many cycles of cell division. Dr Yang discovered

that 12% of these patients display genomic rearrangements near a DNA region known as a promoter, which controls the activity of the TERT gene. In glioblastoma multiforme, an aggressive form of brain cancer, a more complex picture is emerging: structural changes of many different kinds can cause the activation of several key oncogenes

functional consequence of highly clustered genomic rearrangements in cancer is the switching on of oncogenes.

A COMPLEX PICTURE Dr Yang’s research highlights just how intricate the interacting factors influencing DNA changes can be, with multiple mechanisms of mutation sometimes acting to cause rearrangements within one gene, while at the same time multiple driving forces acting on completely separate parts of the genome can together lead to an increased risk of cancer. It seems common that a positive feedback loop is present, where existing mutations

Increasingly, cancers are being linked to genetic causes and inactivation of important tumour suppressors. In fact, says Dr Yang, his research has shown that the main

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The genome view of somatic alterations in a colorectal cancer patient. From the outer rings to the inner ones: chromosomes 1 to Y, base substitutions, copy number changes (red - amplifications, blue - losses), intra-chromosomal rearrangements, inter-chromosomal rearrangements

in genes involved in DNA replication or repair result in an increased likelihood of further mutations including large structural rearrangements. Dr Yang has shown that almost all cancer patients have at least one alteration in a gene involved in DNA replication or repair, and almost half of these are themselves the result of structural rearrangements to the genome.

exome sequencing data are typically only used to discover single base substitutions. Dr Yang has also improved his approach to scour exome data for structural rearrangements to re-use the vast amount of existing data, which has yielded further valuable findings.

approach in which Dr Yang specialises.

Dr Yang is quick to point out that this is an emerging field, and an inexact science. “The rearrangements we observe” he comments, “are a snapshot … of the alterations in cancer genomes.”

His research group at the University of Chicago is, he says, “battling on the frontiers of precision medicine.” Precise characterisation of the mechanisms and nature of genomic rearrangements is crucial: by understanding the genetic mechanisms underlying tumour development, we come one step closer to discovering new drug targets, and ultimately, to helping patients.

Almost all cancer patients have at least one alteration in a gene involved in DNA replication or repair

While wholegenome-sequencing data, which offers unprecedented accuracy and resolution, is the ‘holy grail’ for researchers trying to identify DNA rearrangements, the time and expense needed to generate it remains a constraint. Much more widely available are so-called ‘whole exome sequences’ focusing only on the DNA regions thought to code for protein – known as ‘exons.’ The whole

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On the other hand, although the greatest interest in structural rearrangements so far has been in those within genes themselves, there is growing evidence that changes in DNA regions previously considered ‘non-coding’ may also play an important, though perhaps more subtle, role. These changes might only be picked up through the whole genome

Behind the Bench Dr Lixing Yang E: lixingyang@uchicago.edu

Research Objectives The goal of Dr Yang’s research is to understand the roles of genomic rearrangements in human cancer by development and application of novel computational methods. In particular, he focuses on analysis of whole-genome sequencing data, which offers unprecedented accuracy and resolution in identifying such rearrangements.

Funding National Cancer Institute (NCI)

Q&A

How did you first become interested in this area of cancer research?

Cancer is a disease caused mostly by genetic and epigenetic alterations.

Cancer research not only can help millions of patients, but can also advance our understanding on many fundamental processes of biology including organ development, various signalling pathways, metabolism, cell– cell interactions, ageing, immunology, and many others.

How do structural rearrangements of the genome cause cancer?

The best-known cases are gene fusions – two different genes normally far away are fused into one new gene and the new gene performs a novel function in the cells. There are certainly other ways such as the relocation of regulatory

T: +1 773 834 2948 W: http://yanglab.me

Collaborators • The Cancer Genome Atlas (TCGA) Research Network • International Cancer Genome Consortium (ICGC) Bio Dr Yang completed his PhD in plant genome evolution at the University of Georgia. He joined the Park lab at Harvard Medical School in 2010 before starting his own research group at the University of Chicago in January 2017, continuing his work on cancer genomics.

elements altering the gene functions that they target.

What are the easiest types of rearrangements to identify? And the hardest?

The type doesn’t matter too much; what matters is where they are. There are a lot of DNA fragments that have been repeated many times in the human genome and some copies are completely identical. The ones in such regions are the hardest to identify. The situation is more like being given two pictures: The Empire State Building and a random single family house. Which one is easier to identify?

Does your research suggest that many cancers are heritable? Or that they result from mutations occurring during our lifetime? Or both?

There are certainly some inherited variants which increase the risk of cancer,

Contact Lixing Yang, PhD Assistant Professor Ben May Department for Cancer Research Department of Human Genetics Institute for Genomics & Systems Biology Comprehensive Cancer Center The University of Chicago 929 E. 57th Street, GCIS W432 Chicago, IL 60637 USA

but most cancers are not heritable.

There are damages occurring to DNA all the time and the vast majority are repaired. Only a very small number accumulate during each cell cycle.

Over the years, more and more mutations accumulate in different cells.

Once the mutations reach a certain combination, cancer starts to develop.

How do you think your findings could be translated into treatments for cancer?

Many targeted therapies for cancer block protein functions of fusion genes.

If we discover more rearrangement events that are responsible for tumourigenesis, some of the genes involved in the rearrangements could be good drug targets.

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Health & Medicine ︱ Dr Richard Koya

Reprogramming the immune system for personalised immunotherapy against cancer For decades, researchers have strived to understand how the immune system recognises and fights cancer, ultimately aiming to exploit and augment these processes to create more effective cancer therapies.

Dr Richard Koya, Associate Professor of Oncology, Associate Director of the Center for Immunotherapy, and Director of the Vector Development & Production Facility at Roswell Park Cancer Institute is a prominent researcher in this area. He leads an international team of researchers in developing cutting edge technologies to use patients’ own immune systems to target and kill cancer cells.

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ur immune system is not only critical in our defence against bacteria, viruses and other pathogens, but it also plays an instrumental role in preventing and fighting cancer. The immune response to cancer is complex, involving antibodies and a broad repertoire of white blood cell types, including lymphocytes and other leukocytes.

Cancer immunotherapy refers to any strategy that uses the immune system to fight cancer, and several breakthroughs have brought immunotherapy to the forefront of cancer treatment in recent years. Immunotherapy offers several advantages over conventional chemotherapy, including reduced side effects and the potential to suppress cancer reoccurrence through the

Several T cell subsets exist, each with their own unique TCR and specialised function.

Cytotoxic T cells and T helper (Th) cells are characterised by the presence of a CD8 or CD4 receptor on their cell surfaces, respectively. Cytotoxic T cells become activated when they recognise pathogens or tumour cells via their TCR. This is followed by the release of enzymes and toxic proteins that trigger programmed cell death in target cells. Th cells, as their name suggests, help other immune cells by releasing important chemical messengers called cytokines, which refine and regulate immune responses.

THE IMMUNE SYSTEM VS. CANCER Continuous advances in the understanding of cancer immunology pave the way for significant clinical

While cancer vaccines represent an enticing strategy, the presence of immune resistance mechanisms in cancer cells can limit their long-term efficacy development of long-lasting immune responses.

RECEIVING THE CANCER SIGNAL – THE T CELL RECEPTOR One class of white blood cells, known as T cells, has long been known to play an important role in the immune response to cancer. T cells harbour signalling proteins called T-cell receptors (TCR) on their surfaces, a feature that distinguishes them from other white blood cells. In a complex process, TCRs recognise pathogen- or tumour-specific proteins (known as antigens) that are presented by other immune cells and also cancer cells.

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translation of novel immunotherapeutic strategies. Cancer vaccines were among the first immunotherapies to emerge in the early 1980s, with the FDA-approved hepatitis B virus (HBV) vaccines developed to prevent chronic HBV-induced liver cancer. In simple terms, cancer vaccines are created using specific tumour antigens (TA) – stimulating the immune system to mount a targeted anti-tumour response.

While cancer vaccines represent an enticing strategy, the presence of immune resistance mechanisms in cancer cells can limit the long-term efficacy of vaccination.

Despite intense efforts in cancer vaccine development, progress has been slow in

Patient B

Patient A

Harvest T cells from new patient

Tumour Human Tumour

Subcloning of anti-cancer TCR into a viral vector

MHC I TCR

Cancer-specific T cell

Cancer-redirected T cell

T-cell receptor (TCR)-engineered T cells for metastatic cancer treatment Take the TCR genes from one patient who beat cancer and use them to engineer a cancer-fighting immune system in other patients

Take the TCR genes from one patient who beat cancer and use them to

this area. To date, no cancer vaccines have been approved for use in all individuals by both the American Food and Drug Administration (FDA) and the European Medicines Agency (EMA).

ADOPTIVE CELLULAR THERAPY While efforts to develop cancer vaccines continue, researchers including Dr Koya are continuously exploring other strategies. One such strategy is based on adoptive cell transfer (ACT). This is a form of personalised immunotherapy whereby a small number of T cells are taken from a patient’s blood or tumour, sorted and multiplied in vitro (in the laboratory), and then reinfused into the same patient’s blood – in a similar manner to a blood transfusion. ACT usually follows a period of pre-conditioning chemotherapy known as lymphodepletion, where patients’ overall lymphocyte numbers are reduced to maximise the chances of proliferation of the newly infused T cells.

The potential of ACT was first demonstrated by the National Cancer Institute (US) following long-standing

observations that the presence of tumour infiltrating lymphocytes (TILs) is associated with improved clinical outcomes in a broad range of cancers.

TILs refer to any white blood cells that leave the bloodstream and migrate into a tumour. While early ACT attempts using in vitro-selected TILs seemed promising, methods for isolating and processing TILs are labour-intensive and only benefit a subset of cancer patients.

TCR-ENGINEERED T CELL IMMUNOTHERAPY – IT’S IN THE GENES! For more than a decade, Dr Koya has focused on advancing ACT for highly specific and sustained cancer treatment.

His research centres specifically on TCREngineered T Cell Immunotherapy, using genetic engineering to reprogramme or ‘re-educate T cells to deliver a lethal hit on malignant cells in an extremely precise and efficient way’.

Dr Koya’s group utilises viral DNA to inject TA-specific TCRs into T cells. These newly inserted receptors then target

these cells to the tumour in a highly specific manner. This approach improves ACT, because the engineered T cells are superior in targeting tumours than the in vitro-selected TILS used in earlier ACT attempts. In other words, T cells are reprogrammed specifically to destroy cancer cells. By expanding these cells under specialised laboratory conditions, billions of highly active tumour-specific T cells can be injected back into the patient, targeting tumours with high efficiency and specificity.

Engineered T cell ACT has the potential to target any kind of cancer, but the most widely studied cancer to date is metastatic melanoma.

FROM LAB TO CLINIC Previous studies performed under Dr Koya’s guidance, and indeed by other research groups, have revealed dramatic clinical responses with ACT therapy using TCR-engineered T cells in patients with metastatic melanoma. However, while response rates often exceed 50%, relapse is eventually seen in most patients. Dr

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TGFβ Blockade in TCR-Engineered T cell Ca

for Advanced Solid Tumo

Clinical Trial: Roswell Park Cancer Institute

Clinical Trial: Roswell Park Cancer Institute TGFβ blockade in TCR-engineered cancer immunotherapy for advancedTsolid TGFβT cell Blockade in TCR-Engineered celltumours Cancer Immunotherapy

Clinical grade (cGMP) production of the retrovirus vector and testing in human T cells tetramer NY-ESO1 TCR

Clinical grade (cGMP) production of retrovirus vector testing in human T cells forthe Advanced Solidand Tumors NY-ESO1 TCR

tetramer NY-ESO1 TCR

Clinical grade (cGMP) production of the retrovirus vector and testing in human T cells NY-ESO1 TCR

LTR LTR

Ψ Ψ

TCRα P2A TCRβ T2A dnTGFβRII TCRα P2A TCRβ T2A dnTGFβRII

LTR

LTR

dnTGFβRII

dnTGFβRII

dnTGFβRII

dnTGFβRII

Phase I/IIa Clinical for Advanced NY-ESO1(+) Cancer (accruing patients) 5. Phase I/IIa Clinical Trial Trial for Advanced StageStage NY-ESO1(+) Cancer (accruing patients)

5. Phase I/IIa Clinical Trial for Advanced Stage NY-ESO1(+) Cancer (accruing patie T cells Leukapheresis

Retrovirus

Activation (OKT3+IL-2)T Leukapheresis

cells LTR

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TCRα P2A TCRβ T2A dnTGFβRII

LTR

RV-NYESO1-TCR/dnTGFβRII Activation transduction

(OKT3+IL-2)

Cyclophosph. conditioning

LTR

FDA Approved 5/17 (IND 17410) Retrovirus

release test test Lot release

Ψ

TCRα P2A TCRβ T2A dnTGFβRII ACT (T cells)

LTR

RV-NYESO1-TCR/dnTGFβRII 0 1 7 14 30 transduction

R01 NCI/NIH CA164333 (PI: Richard Koya)

PBMC collections

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FDA Appro (IND 17410

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FDG PET or CT (screening, wk. 4, 12) Tumour Biops./TIL harvest (screening, wk. 4, upon progression)

TCR-engineering, because T cells are Koya’s group hypothesises that the highly also engineered to lose sensitivity to the active TCR-engineered T cells eventually Cyclophosph. immunosuppressive effects of secreted succumb to the immunosuppressive conditioning TGF-β. If the hypothesis is correct, these tumour microenvironment – an specially engineered T cells should exhibit environment that is imposed largely by the secretion of a transforming growth factor beta (TGF -β) R01 CA164333 (PI: Richard Koya) by tumour cells.NCI/NIH This hypothesis is supported

by Dr Koya’s previous clinical trial experiences, sustained efficacy in the microtumour and is currently being tested in a new environment, resulting in fewer cases of National Institutes of Health-funded relapse, and better patient outcomes. prospective clinical trial at Roswell Park Cancer Institute, led by Dr Koya.

Previous clinical trials based on ACT have mainly focused on the use of cytotoxic T The trial will specifically address a novel cells only. However, evidence is growing ACT approach based on an immunefor the efficacy of Th cells in ACT. Since modulator-enhanced TCR-engineered T Th cells are now known to promote the cell transfer for metastatic cancer patients. maintenance of cytotoxic T cell responses This investigation takes TCR-engineered against cancer cells, as well as rescuing T cell transfer one step further than

exhausted T cells, it is anticipated that sustained immune responses can be realised by the synergy of CD8TCR- and PBMC collections CD4TCR-engineered T cells. This idea is likely be investigated in future FDGtoPET or CT (screening, wk. 4, 12) clinical trials with ACT. Tumour Biops./TIL harvest (screening, Another avenue under upon progression) investigation is the combination of ACT with therapies that block other cellular pathways used by cancer cells to evade the immune system.

Dr Koya’s research uses genetic engineering to re-educate T cells to deliver a lethal hit on malignant cells

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Immunotherapy is an exciting area, and new approaches currently dominate ongoing clinical trials in the pursuit of effective, personalised cancer treatments.

These therapies might one day become the standard of care for cancer patients.

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Behind the Bench Dr Richard Koya

E: richard.koya@roswellpark.org T: +1 (716) 845 1300, ext. 6582 W: www.roswellpark.edu/richard-koya W: http://tactivatherapeutics.com/ W: www.youtube.com/watch?v=VwUJHR-Wznc W: https://clinicaltrials.gov/ct2/show/NCT02650986?term=koya&rank=1

Research Objectives Dr Koya’s research focuses on developing innovative ways to utilise the patients’ own immune system to target and kill cancer cells.

His approach is based on genetic engineering of a subset of immune cells called T lymphocytes, by utilising viral vectors to re-programme these immune cells.

Funding National Institutes of Health (NIH) Collaborators • Dr Kunle Odunsi, Deputy Director of Roswell Park Cancer Institute

Q&A

What has been the biggest technical challenge in your research to date?

The biggest challenge in my research was to select among many candidates the most efficient and more specific TCR to target a Tumour Associated Antigen that is in cancer but not in normal tissues.

Do you believe that future developments in immunotherapy will one day completely replace conventional chemotherapeutic approaches?

I would not say replace completely since some chemo-agents are also useful to prime the cancer cells, making them more susceptible to immunotherapies.

• Dr Thinle Chodon, Director of Translational Research Operations at the Center for Immunotherapy Bio Dr Koya received his MD in 1991 and completed his residence in Internal Medicine and fellowship in Clinical Oncology at UFRGS, Brazil. He received his PhD in Molecular Biology & Pathology from Hokkaido University in 2001. From 2001–2003 he pursued a postdoctoral fellowship at USC, Los Angeles, before joining UCLA in 2003.

He became Associate Professor of Oncology at RPCI in 2013.

Are there likely to be undesired effects or safety issues with widespread use of ACT? The main issue that sometimes accompanies ACT is the widespread inflammation that can happen in certain patients with certain types of cancer, a side effect called cytokine release syndrome. This is much less common in TCR-based therapies, but much more prevalent with CAR-T cell, another modality of ACT.

Contact Richard C Koya, MD, PhD Associate Professor of Oncology and Immunology Director of the Vector Development & Production Facility Associate Director of the Center for Immunotherapy Roswell Park Cancer Institute Center for Immunotherapy Elm and Carlton Streets, CCC-419 Buffalo, NY, 14263 USA

Do you anticipate that patients can be resubjected to ACT after a relapse?

Yes, and this is also a plan we are pursuing. The point is that in early-phase clinical trials, the FDA recommends a trial with a single dose to first check for side-effects, as safety is paramount.

Are there tumour types that ACT is unlikely to be effective against?

Actually no. Any solid or liquid tumour can be targeted by ACT.

There is no tumour type against which ACT will be ineffective; any solid or liquid tumour can be targeted by ACT

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Health & Medicine ︱ Dr Shoukat Dedhar

Inhibiting cancer stem cell survival in the hostile tumour environment Wouldn’t it be great if a small non-toxic molecule could be used to treat cancer? By investigating the possibility of using a cancer cell’s own physiology as a weapon against it, Dr Shoukat Dedhar, at the University of British Columbia, is developing a new treatment that could help prevent tumour growth and metastasis.

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very two minutes, someone in the UK is told they have cancer, with one out of every two people likely to be diagnosed at some point in their lives.

The disease is the world’s second leading cause of death, and was responsible for nearly nine million global deaths back in 2015.

Current treatment methods involve the removal of cancer cells either through surgery or via radiation or toxic chemicals.

However, cancer cells are tricky, and it is

cancerous cells remain, they can cause a resurgence of the disease. Think of it like an ant’s nest – you can get rid of nearly all of them, but leaving just two or three could be enough for the nest to reappear down the line.

Dr Dedhar and his team are investigating a new type of treatment, which takes advantage of the unique physiology of cancer cells. It aims to inhibit their growth and ability to metastasise, without damaging healthy body cells.

Inhibition of CAIX resulted in significant depletion of cancer stem cells within tumours very difficult to eradicate every single one of them. If the cancer has metastasised (spread from the original tumour to another part of the body), surgery can rarely remove every cell, and treatments that typically kill cancer cells are generally toxic to normal cells as well. If even a few

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The results so far have proved very encouraging.

A HOSTILE ENVIRONMENT To understand how the new treatment works, we must first understand the inner workings of a tumour. Tumours

CAIX-Targeted Therapy is a Weapon used to Exterminate CAIX-Targeted is a Weapon used to Cells Exterminate Hypoxic,Therapy Treatment-Resistant Cancer Hypoxic, Treatment-Resistant Cancer Cells Hypoxic Tumour Hypoxic Tumour

CAIX-Positive Cancer CAIX-Positive Cancer Stem Cells Stem Cells Hypoxic Microenvironment Hypoxic (Cancer Stem CellMicroenvironment Niche and CAIX(Cancer StemHypoxic Cell Niche and CAIXPositive Cancer Cells) Positive Hypoxic Cancer Cells) Blood Vessels Blood Vessels

CAIX-Negative CAIX-Negative Cancer Cells Cancer Cells CAIX-Positive CancerCAIX-Positive CancerAssociated Fibroblasts Associated Fibroblasts

Chemotherapy Chemotherapy Radiotherapy Radiotherapy

Treatment Treatment

CAIXCAIXTargeted Targeted Therapy Therapy

Chemotherapy Chemotherapy Radiotherapy Radiotherapy

Functionally Impairs/Kills Functionally Impairs/Kills Cells • Hypoxic Cancer HypoxicStem Cancer Cells Cells • Cancer Stem Cells Fibroblasts • Cancer Associated • Cancer Associated Fibroblasts

Tumour Recurrence Tumour Recurrence

Tumour Regression Tumour Regression

Metastasis Metastasis

Inhibition of Metastasis Inhibition of Metastasis

Resistance to Resistance to Chemotherapy and Radiotherapy Chemotherapy and Radiotherapy

are made up of millions of cells and, as each one grows, the blood supply required to nourish the rapidly dividing cancer cells with oxygen and nutrients becomes inadequate, causing regions of the tumour to become hypoxic (low in oxygen). Like normal body cells, cancer cells cannot survive without oxygen, so to overcome this, they stabilise a very important protein called HIF-1α (hypoxia inducible factor 1 alpha). This protein mediates the activation of numerous genes vital for the adaptation of cancer cells to the hypoxic environment.

Through HIF-1α, cancer cells in hypoxic regions of the tumour begin producing proteins that trigger the growth of hundreds of capillaries from nearby blood vessels. These new capillaries provide the tumour with additional oxygen and nutrients, whilst

also removing waste products such as carbon dioxide. However, the capillaries are often leaky and deformed, so – although they allow the tumour to grow bigger and more quickly – the hypoxic micro-environments inside the tumour remain.

To compound the problem, hypoxic cancer cells alter the way they produce energy and cellular building blocks, such as proteins and fats, to continue their growth in the absence of oxygen.

This altered metabolism produces

micro-environment present in hypoxic parts of the tumour, they will soon die.

ADAPT OR DIE As part of their adaptive response, cancer cells begin producing a cell membrane-bound protein called carbonic anhydrase IX (CAIX). CAIX converts carbon dioxide from outside the cell into bicarbonate and protons.

The bicarbonate is then transported into the cell to reduce the intra-cellular acidity, providing a survival benefit for these cells. The protons remain outside and contribute to the acidification of the microenvironment.

Creating this acidic environment causes cancer stem cells to divide more rapidly, enhancing their ability to invade healthy tissue and metastasise across the body.

By targeting CAIX, we may be able to overcome resistance to chemotherapy and radiotherapy, tumour recurrence and metastasis acidic waste products that build up inside and outside the cells. If cancer cells cannot adapt to the hostile, acidic

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AN ATTRACTIVE TARGET Since cancer cells become absolutely dependent on CAIX to reduce acidification and thus promote their survival, this requirement becomes their “Achilles heel”, thus making CAIX a promising target for cancer therapy.

CAIX is an attractive target for anticancer therapies for several reasons.

First, it is only produced by cancer cells, so could be inhibited with no harmful effects to normal cells. Second, it is critical for the survival of cancer stem cells and their ability to invade healthy tissue. And third, its position on the outer surface of the cell membrane makes it a relatively easy target for drugs.

CAIX is produced in large amounts by cancer cells that metastasise, and Dr Dedhar has shown that if CAIX expression is depleted, cancer stem cells no longer function properly, blocking metastasis. His study, and others like it, have provided proofof-principle data that CAIX inhibition could provide a therapeutic benefit in treating cancer.

Dr Dedhar believes that by targeting CAIX, we may be able to overcome resistance to chemotherapy and radiotherapy, tumour recurrence and metastasis. He and his team have developed a targeted small molecule inhibitor of CAIX and have shown that

this inhibitor reduces tumour growth and metastasis in models of human cancer.

Inhibition of CAIX in human breast cancer cells in the laboratory prevented breast cancer stem cells from dividing and replenishing the cell population in hypoxia. The team also tested the CAIX inhibitor in a mouse model of breast cancer, with CAIX inhibition resulting in a significant depletion of tumorous cancer stem cells. Combination treatment using the inhibitor with paclitaxel (a chemotherapy drug) was also found to enhance tumour growth delay and completely eradicate metastasis of cancer cells to the lungs, when compared to treatment with paclitaxel alone.

The team have recently completed a phase 1 safety clinical trial of the CAIX inhibitor, with findings showing it to be safe and well tolerated by patients.

They are now embarking on clinical trials in which the CAIX inhibitor can be combined with standard chemotherapy regimens, and Dr Dedhar anticipates significant, additive effects on tumour growth – especially on recurrence and metastasis. Through his team’s tireless work, a cancer diagnosis might not be the death sentence it once was.

Behind the Bench Dr Shoukat Dedhar

E: sdedhar@bccrc.ca T: +1 604 675 8029 W: www.bccrc.ca/ W: https://clinicaltrials.gov/ct2/show/NCT02215850 Research Objectives Dr Dedhar’s research focuses on the role of ILK signalling in cancer progression, the molecular basis and targeting of centrosome clustering in cancer cells, and the therapeutic targeting of tumour hypoxia effectors, Carbonic Anhydrases IX and XII. Funding The Canadian Institutes of Health Research (CIHR)

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Collaborators • Dr Claudiu Supuran • Welichem Biotech, Inc. Bio Dr Dedhar received his BSc (Hons) in Biochemistry from the University in Aberdeen, Scotland, and his PhD from the University of British Columbia, Canada in 1984. He carried out a Postdoctoral Fellowship in the laboratory of Dr Erkki

Ruoslahti at the Burnham Institute, in La Jolla, California, USA. Contact Shoukat Dedhar, PhD BC Cancer Research Centre Rm 3-110 675 West 10th Avenue Vancouver, B.C. V5Z 1L3 Canada

Q&A

How did you first become interested in CAIX? We were carrying out research to identify causative genes in breast cancer metastasis. We discovered that a set of genes were active in tumour cells that had the capacity to form metastases, but were silent in tumours that did not have this capacity. Interestingly, all of these genes are activated by hypoxia and HIF-1, and one of the most active genes was CAIX. We proved that CAIX was indeed required for breast cancer cells to metastasise by depleting CAIX from the cells with metastatic capacity: this depletion resulted in the almost complete loss of metastatic ability.

Could the CAIX inhibitor be used to treat any type of cancer?

Yes, any type of cancer that produces CAIX could be inhibited by the CAIX inhibitor. However, not all cancers produce CAIX, since some types of cancers are not hypoxic. So, the CAIX inhibitor will be used therapeutically only in tumours that produce CAIX. We will have to analyse the patient tumours for CAIX expression before embarking on a therapeutic regime that includes the CAIX inhibitor.

Could the CAIX inhibitor be used on its own to treat cancer without the need for chemotherapy?

Since only a proportion of cancer cells within a tumour produce CAIX (the proportion can vary from 20% to as much as 80%), it is unlikely that CAIX inhibitor monotherapy would be effective in eradicating the tumour. On the other

hand, while both radiation and chemotherapy will effectively kill the majority of tumour cells, they are ineffective in killing tumour cells within the hypoxic niches which should be killed by inhibitors such as the CAIX inhibitor. Our recent, unpublished work also demonstrates that inhibiting CAIX makes chemotherapy agents such as temozolomide and gemcitabine, more effective.

While we do not understand the reason for this, it is likely that inhibiting CAIX changes the tumour physiology in such a way as to make the tumour cells more sensitive to the chemotherapy agents.

Can you describe the mouse breast cancer model that was used to test the CAIX inhibitor?

We have used several mouse tumour models to test the CAIX inhibitor. Our initial model was transplantation of breast cancer cells into the mammary fat pad of the mouse. This results in the formation of a breast tumour within the mammary gland of the mouse. Once the tumour reached a reasonable size so that it could be palpated, we started administering the CAIX inhibitor orally on a daily basis for approximately 21 days. Control mice received only the “vehicle” used to dissolve the CAIX inhibitor. Tumour volumes were measured daily and the results tabulated. We had also “tagged” the tumour cells so that they could be followed in the mouse using an imaging instrument called IVIS. This allowed us to determine the extent of spread, or “metastasis”, of the tumour in the control and treated mice

What will the clinical trial of the CAIX inhibitor involve?

Having completed the Phase 1 Safety trial with the CAIX inhibitor, we can now embark on clinical trials to test the efficacy of the compound.

Our next trial will be a multicentre Phase 1b/Phase 2 trial in patients with CAIXpositive pancreatic ductal adenocarcinoma, or PDAC. These cancers are very difficult to treat with current therapeutic regimes, and we have found that PDAC patient tumours are very hypoxic and about 40–60% of the cells produce CAIX. Furthermore, our pre-clinical data in mouse models of PDAC demonstrate that adding the CAIX inhibitor to the standard of care therapeutic, gemcitabine, results in substantial survival benefit, with depletion of cancer stem cells. Thus, the clinical trial will involve treatment of CAIX-positive patients with a combination of gemcitabine and CAIX inhibitor. We will measure a number of parameters, including circulating drug levels, and whether cancer stem cells are depleted in the tumours, but a major end point will be to determine whether there was any benefit in progression-free survival.

Similar trials are in the planning stages for “triple-negative“ breast cancer and glioblastoma (brain cancer).

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Health & Medicine ︱ Dr Tania Roth

The epigenetic effects of adverse early-life experiences Environmental factors interact with genetics in driving living organisms’ development throughout life. Epigenetics is the field of study that explores this interaction, as well as its potential effects on individuals’ behaviour and health. Dr Tania Roth, working at the University of Delaware in Newark, USA, carried out extensive research exploring the impact of the environment, particularly stress factors, on individual genes, and how these changes might influence behaviour and psychological development. Her findings indicate that adverse early-life experiences can result in particular alterations in the brain that have consequences for gene expression and behaviour.

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ife experiences and environmental factors play a substantial role in how individuals develop over time, in terms of their behaviour, as well as their physical and mental health.

However, different people who have similar experiences might react to these in entirely different ways and be affected differently. This is often thought to be due to individual DNA and genetic factors.

DNA Methylation

Over the past few decades, a considerable body of research has been investigating the way in which environmental and genetic factors interact with one another, resulting in different effects on individuals’ health, behaviour, and psychological development.

EPIGENETICS: HOW GENES AND ENVIRONMENT INTERACT Epigenetics is the field of research that focuses on the biological changes occurring in living organisms as a result of genes interacting with environmental factors and life experiences. A number of factors can influence a human being’s psychological development and health over time, including prenatal environment, parenting, and other stress encountered throughout life. All these different factors leave their marks on an individual being’s genetic make-up (DNA), in the form of chemical tags that instruct genes on what to do. Studying and observing these alterations could help determine how stress factors can affect different individuals’ physical and mental health. Dr Tania Roth has been investigating the epigenetic alterations in the brain that can occur after early-life adversities, such as maltreatment or poor parenting.

BRAIN ALTERATIONS AFTER ADVERSE EARLY-LIFE EXPERIENCES Dr Roth and her team of researchers carried out experiments on rats to test the epigenetic effects of early-life adversity in the brain, as well as their behavioural outcomes. By providing scarce living resources, such as nesting

Histone modifications Epigenetic marking of the genome by DNA methylation (red) and histone modifications (green and yellow), such as acetylation.

The pups of maltreated female rats showed epigenetic alterations later in life, which still occurred even if the pup was placed with a foster mother materials, they prompted rats to offer poor caregiving to their neonate pups.

They then tested the rodents who received maltreatment for any changes or abnormalities in the brain.

Dr Roth’s research suggested that infant– caregiver interactions could mark genes that have previously been found to play a prominent role in psychiatric disorders.

The rats that received poor caregiving presented a number of epigenetic alterations, which varied according to the rats’ sex. These alterations were particularly pronounced in brain regions known to be affected by child abuse and neglect, such as the prefrontal cortex, the amygdala and the hippocampus.

Maltreatment appeared to cause changes in methylation, the process by which methyl groups are added to the DNA, particularly in neurons. Methyl groups are chemicals made up of a single carbon and three hydrogen atoms.

The biochemical process of methylation is involved in regulating the expression of genes.

The prefrontal cortex is a region of the brain associated with a number of complex behaviours and cognitive functions, while the amygdala is involved in the experiencing of emotions. The hippocampus has been found to play a role in memory and emotion processing.

EPIGENETIC EFFECTS ON BEHAVIOUR Dr Roth’s research found that poor caregiving for neonate rats did not affect their behaviour as adolescents, but significantly affected their behaviour as adults. Female rats who were

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Early life experiences can cause epigenetic changes that affect behaviour into adult life

maltreated as pups appeared to be more anxious days before giving birth and also displayed the same types of adverse caregiving behaviour towards their offspring that they received from their caregivers in early-life. Moreover, the pups of maltreated female rats also showed epigenetic alterations later in life, and some of these alterations still occurred even if the pup was placed with a foster mother who had not been maltreated in early life.

As well as displaying these disruptions in maternal behaviour, the maltreated females also performed poorly in a series of cognitive tasks. The male rats tested during the study appeared to present problems with extinguishing fear-related or troubling memories.

PREVENTING OR REVERSING EPIGENETIC STATES Understanding the epigenetic alterations that follow adverse early-life experiences or environmental factors could help to devise policies or treatments that could either prevent or reverse these changes in the brain.

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The rats that received poor caregiving presented a number of epigenetic alterations, which varied according to the rats’ sex Dr Roth’s research has found that treating adult animals with a drug that changes methylation states can somehow counteract or rescue the changes in DNA methylation and gene expression produced by the maltreatment. She is now exploring the possibility that the same pharmacological treatment that could reverse these epigenetic states might also be able to affect the maternal behaviour of maltreated females, leading them to display more nurturing caregiving tendencies.

In future, a similar treatment could be developed to counteract the effects of poor caregiving on methylation states earlier on in development, which might change the behavioural trajectories that follow mistreatment and prevent the adult rats from replicating the same dysfunctional behaviour while raising their own pups.

A MEANINGFUL CONTRIBUTION Dr Roth’s research has provided evidence suggesting that maltreatment and other adverse environmental factors in early life can produce changes in methylation processes within the brain, which result in altered gene activity and adult behaviour. Her work has substantially contributed to the field of research exploring epigenetics, providing further empirical evidence of preventable and potentially reversible epigenetic states.

If Dr Roth’s observations were found to be similar in humans, her work could aid the development of pharmacological or behavioural treatments to prevent, counteract and reverse some of the behavioural and developmental effects of adverse early-life experiences.

Behind the Bench Dr Tania Roth

E: troth@psych.udel.edu T: + 1 302 831 2787 W: http://rothlab.psych.udel.edu/

Research Objectives Dr Roth’s work explores the impact of the environment, specifically stress, on our genes and how these epigenetic changes influence our behaviour and development.

Funding • National Institutes for Health (NIH) Collaborators • Dr Mary Dozier, University of Delaware

Q&A

When did you first become interested in the interaction of environmental factors and genetics, as well as the effect it might have on human health and development?

During my graduate training I became interested in understanding how an experience or environmental factor could get under the skin to influence development and behaviour. I was fascinated by the brain’s capacity to change because of experience, and curious how factors like child abuse and neglect could hijack developmental processes or have such long-lived consequences for behaviour and mental health.

What are the greatest challenges of studying epigenetic effects on health and development?

Development is a lifelong process, so to truly understand epigenetic effects on health and development studies you need a longitudinal design. But gathering data, especially epigenetics data, from the same subjects repeatedly over a period of

Bio Dr Tania Roth is an Associate Professor and Director of Graduate Education in Psychological and Brain Sciences at the University of Delaware, where she teaches upper-level and graduate courses in psychology and neuroscience. Her research programme is focused on defining epigenetic mechanisms responsible for environmental influences on gene activity, development of behaviour, and psychiatric disorder.

Contact Tania L. Roth, PhD Associate Professor Psychological and Brain Sciences University of Delaware 108 Wolf Hall Newark, DE 19716 USA

time is very difficult. Identifying whether an epigenetic change is coincidental or causal in relation to an outcome is another challenge but necessary for us to really understand epigenetic effects on health and development.

as well and what evidence is there supporting this?

Yes. Several reports indicate similar methylation changes in humans with a history of maltreatment. Specifically, they see the same change in DNA methylation in peripheral measures that we see in the brains of our rodents. There is also some evidence that certain forms of behavioural therapy and intervention can change epigenetic states in humans.

What do you feel are the most important findings you have collected so far through your research and why?

We have shown that poor parenting early in development can have multigenerational consequences on DNA methylation states in the brain.

These data provide an empirical basis for the far-reaching effects of early adversity, helping us understand some of the biology of how factors like child abuse and neglect can affect health and development. But we have also shown that epigenetic manipulations can reverse these states, and projects underway in the laboratory suggest these strategies also rescue aberrant behaviour. Such data demonstrate that the epigenome retains malleability throughout life and suggest it could be a target for changing brain and mental health.

Do you believe your findings on rodents could be applied to humans

What are your plans for further research and investigation?

We are working to translate our findings to humans. In current collaborative research with Dr Mary Dozier (a clinical psychologist at the University of Delaware), we are studying DNA methylation patterns in parent–child dyads who were involved with Child Protective Services as the result of allegations of neglect. We are also investigating the reversibility of these epigenetic alterations following a behavioural intervention programme (designed by Dr Dozier) that is based on attachment theory and stress neurobiology.

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Thought Leader

IAPB: Envisioning the future of universal eye care Everyone across the globe needs access to the best possible standard of eye health as advocated by the International Agency for the Prevention of Blindness (IAPB). IAPB was established as a coordinating umbrella organisation to lead international efforts in blindness prevention activities and eye health. Research Outreach spoke to CEO Peter Ackland who discusses IAPB’s mission, the importance of World Sight Day, IAPB’s relationship with the World Health Organization (WHO) and more in greater detail.

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ision really matters as it is vital to our everyday lives. Naturally, sight is the main sense that people fear losing the most. According to the latest data in 2015, 253 million people are estimated to be visually impaired worldwide: 36 million of whom are blind and 217 million with severe or moderate vision impairment. Another 1.1 billion people are estimated to have near-vision impairment. Yet globally, a majority of all visual impairment can be prevented or cured. No one understands this more than the International Agency for the Prevention of Blindness (IAPB).

IAPB is an alliance of civil society organisations, corporates and professional bodies promoting eye health through advocacy, knowledge and partnerships.

IAPB’s mission is to eliminate the main causes of avoidable blindness and visual impairment by bringing together governments and non-governmental agencies to facilitate the planning, development and implementation of sustainable national eye care programmes. We managed to speak

Hi Peter! Can you tell us a bit more about your role as CEO of IAPB and the role of IAPB as an organisation?

I have been the CEO of the IAPB for the past nine years. The most important part of my job is external representation of the organisation and creating partnerships that can help IAPB achieve its mission – to eliminate avoidable blindness and visual impairment. IAPB is a global membership organisation.

All major international not-for-profit organisations working to improve eye health are our members and an increasing number of national level eye hospitals, research institutions, foundations and NGOs. Following on from the previous question, can you tell us briefly about the IAPB’s core principles, heritage and mission as well as its impact?

IAPB encourages membership from as broad a mix of stakeholders as possible because we think everyone makes a vital contribution to achieving our vision of a world free of avoidable blindness.

IAPB’s key areas of work are advocacy as well as promoting good practice

passing of four World Health Assembly Resolutions on avoidable blindness this century – great recognition of the importance of better eye health by the apex health organisation (WHO).

Another area of advocacy success has been working with major donors such as Standard Chartered Bank and the Queen Elizabeth Diamond Jubilee Trust, who have both chosen the elimination of avoidable blindness as their main philanthropic programmes and together have committed around $200m to eye health projects.

Can you tell us about the IAPB’s collaboration with the World Health Organization (WHO) and their impact on the IAPB’s goals?

IAPB is in official relations with the WHO. That means we collaborate to promote better eye health. WHO is accountable to countries, whilst leading on the development of policy and guidelines to promote better health.

IAPB and WHO launched VISION 2020: The Right to Sight, a global initiative in 1999 and this has had significant success

Globally there are an estimated 253 million people with serious levels of vision loss that substantially impact on their lives with CEO Peter Ackland who discussed with us the role of the organisation and their strategy for the future to help improve eye health worldwide.

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in the development of eye health services. IAPB is an alliance, so IAPB staff work closely with our members to deliver our advocacy objectives. Some notable advocacy successes include the

in bringing many actors working in the field of eye health together. The four World Health Assembly resolutions have consolidated that understanding, culminating in the Global Action Plan

(GAP) for 2014–2019. IAPB’s current work with WHO is mainly focused upon the implementation of the GAP, particularly at country level. At the most recent WHA in May 2017, IAPB and member countries called for WHO to produce a “World Report on Vision”. Later, this was adopted as an action by all countries in the full Assembly. WHO intend to produce the World Report on Vision in time for World Sight Day 2018. The report will lay down the agenda over the next decade for promoting better eye health for all. IAPB is supporting the preparation of the report with funding and providing our inputs.

IAPB works with a variety of WHO departments – disability, health systems, the health workforce, health management information and ageing – apart from its Prevention of Blindness unit.

Can you tell us a bit about this year’s

‘World Sight Day’ and its importance for IAPB? How does WSD differ from the other global health initiatives at IAPB? World Sight Day (WSD) always falls on the second Thursday of October each year. It is our major health day of the year to raise awareness of avoidable blindness and visual impairment and the need for better eye health.

This year’s WSD focuses on the launch of our latest edition of the IAPB Vision Atlas.

We are anticipating that our colleagues and Members in every part of the world will be using WSD to promote several key advocacy messages – using the Vision Atlas data about their country to evidence their advocacy ask.

We will also be talking about three major threats for the future – the increasing global population and the rapidly increasing ageing of that population, the increasing number of people with

diabetes and the very concerning increase in people with myopia.

Collectively, it is estimated that, by 2050, the number of people who are blind or have visual impairment could increase from today’s 253m to more than 700m – almost a threefold increase.

Improving eye health services and making them accessible to all needs emphasis, particularly for those who are most vulnerable for whom services must be free at the time of access.

Can you explain the role of the IAPB Vision Atlas, its importance and how it is used at IAPB? The Vision Atlas is one of IAPB’s flagship projects. It is available in two forms – online and as a paper publication. We see it as the first point of reference for anyone who wants to know the latest numbers, evidence and issues relating to eye health.

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©IAPB

The Vison Atlas is built around two main data sets. The first is the Vision Loss Expert Group (VLEG) estimates for global, regional and country level prevalence and causes of visual impairment and blindness. A series of country level maps help visualise this data. By hovering over a country you can ascertain prevalence disaggregated by level of impairment, sex, age and over time from 1990 through to 2020.

21 Regional maps based on the Global Burden of Disease studies show the causes and the change over time.

Several articles bring the data to life and highlight key issues. The major risks for the future are looked at as well as the opportunities – we could for example eliminate blindness from two very ancient diseases, trachoma and river blindness.

The second data set is IAPB’s country indicator dataset. The WHO recommends countries collect Global Action Plan identified indicators to measure the state of their eye health services and to monitor change. IAPB has collected the latest data available for these indicators for 191 countries.

Again, this is all easily accessible and supplemented by articles. There is a focus on the numbers of trained eye health workforce at country level and their chronic shortage in many poor countries, especially in Africa.

Finally, the Vison Atlas contains many resources and other information such as the common eye conditions and their impact on sight loss.

From your website, the IAPB clearly has some major sponsors supporting them, from Sightsavers to the Queen Elizabeth Diamond Jubilee Trust – how do you attract new supporters and which organisations do you hope to work with in the future to work towards the elimination of avoidable blindness?

Sightsavers and the Queen Elizabeth Diamond Jubilee Trust are just two of our 150+ Members – all are equally important to us. IAPB is mainly funded from membership fees and additional grants that some of our members pay to support particular projects.

A big focus of IAPB‘s work is to promote collaboration so that our members can

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learn from each other and build upon each other’s work. This not only mitigates against re-inventing the wheel but the collective effort is greater than the individual parts. We are always looking for new members to join. We are seeing a welcome trend; an increasing number of new members work at the national level.

We also aim at making links with organisations beyond the eye health world. I see us becoming very close with

those working in the health workforce, ageing, diabetes and disability as particularly important going forward.

What are the highlights from your recently published 2016 annual report?

Did IAPB achieve it’s set goals?

A major highlight in 2016 was IAPB’s 10th General Assembly, held in Durban.

10GA brought together 1,150 eye care professionals from 100 countries.

©IAPB

Thought Leader

The President of Liberia (and Nobel Peace Prize winner), Ellen Johnson Sirleaf, graced the opening ceremony along with the South African Minister of Health, Dr Aaron Motsoaledi. Symposium speakers included Dr Matshidiso Moeti, Regional Director, WHO Africa, Mr David Donoghue, Irish Ambassador to the UN (and Chair of the final stages of the UN meetings that developed the Sustainable Development Goals), Dr Tim Evans from the World Bank and Dr Francis Omaswa, of the African Centre for Global Health and Social Transformation. A talk from Professor Astrid Stuckelberger on healthy ageing was a highlight. There were 42 courses covering a range of eye health and related topics. We had over 60 sessions with 200 speakers and 250 poster presentations over the three days.

On IAPB’s website, goals are outlined from 2013–17. What are the future goals for IAPB in the efforts to achieve universal eye health?

The most important objective for IAPB and our members going forward is the need to achieve greater priority at country level for eye health. International resolutions are important but ultimately it is what individual governments do that is key. We know most avoidable blindness and visual impairment is found within the poorest people in the poorest countries. Giving these people access to eye health services is the key to making sure that the horrifying estimates for the future made by the VLEG (700m by 2050) are not realised.

Much greater emphasis must be placed on improving eye health services and making sure they are accessible to all Improving access requires more trained eye health staff at primary, secondary and tertiary level and more and better equipped hospitals and eye clinics. It requires more work at community level and public eye health messaging and it needs increased domestic funding in eye health provision. Above all the poorest need to be protected against out of pocket payments, arguably the single biggest obstacle to poor people gaining

the treatments they need, through the provision of targeted social insurance schemes or government funded services through taxation.

• For more information about the IAPB, membership and World Sight Day, please visit the website at https://www.iapb.org/

Contact

The International Agency for the Prevention of Blindness (IAPB) London School of Hygiene and Tropical Medicine Keppel Street, London WC1E 7HT United Kingdom E: communications@iapb.org W: http://www.iapb.org/

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Health & Medicine ︹Dr Warren Grundfest and Dr Zachary Taylor

A new terahertz medical imaging tool could provide early detection of corneal disease Recent research in medical imaging found that tools based on terahertz (THz) frequency illumination could help map the distribution and movement of water near the surface of body tissues. Dr Warren Grundfest and Dr Zachary Taylor, at The UCLA Henry Samueli School of Engineering and Applied Sciences, are developing a new imaging tool for non-contact high-resolution measurements of corneal hydration. This could be a promising method to accurately detect and study cornea-related diseases and pathologies, such as Fuchs’ endothelial dystrophy, intraocular lens implantation complications and corneal graft rejection.

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erahertz (THz) imaging is an emerging non-destructive scanning and imaging technique used within several settings, ranging from pharmaceutical and biomedical applications, to integrations in tools for security and aerospace industries. Ongoing research in THz for medical imaging has revealed that THz frequency illumination may be ideal for mapping the distribution and movement of water in physiological tissues. This might be particularly useful for quantifying the water content within the corneal tissue which is generally much less heterogenous than other tissue systems in the human body.

A group of scientists, engineers and clinicians in California are investigating this possibility, trying to develop an imaging tool that can provide accurate measurements of corneal hydration, to help detect and study corneal disorders.

Right: The cornea forms the outer most layer of your eye. It is approximately 79% water by volume. Many diseases can perturb the endothelium which leads to changes in stromal water content. These changes are hard to detect in the early stages.

IMAGING CORNEAL DISORDERS Corneal disorders, such as Fuchs’ endothelial dystrophy, affect a considerable number of people worldwide – particularly within the elderly population. They often result in chronic vision impairment and commonly those affected require surgical intervention. One of the key physiological problems behind these disorders is believed to be an increase in hydration of the corneal tissue, referred to as Corneal Tissue Water Content (CTWC). In fact, many corneal disorders are characterised by an abnormally high CTWC, which causes a swelling of the cornea and subsequent blurring of vision. This makes CTWC an important diagnostic target for doctors and physicians looking to diagnose corneal disorders: not only does it allow the disease to be observed, but it also indicates potential tissue damage. Until now there have been no efficient noninvasive ways of accurately and directly measuring CTWC in situ. New research conducted by the Californian researchers aims to change this.

Dr Taylor and Dr Grundfest are two of these researchers working to develop a non-invasive imaging tool based on THz frequency illumination, which could help measure the hydration levels of the cornea’s tissue. Current methods of measuring CTWC are based on ultrasonic or optical thickness measurements, which can be measured very accurately. However, mapping the cornea’s hydration using these thickness measurements can be extremely inaccurate. Dr Taylor and Dr Grundfest’s newly-developed imaging system would allow scientists and clinicians to acquire spatio-temporal variation maps of CTWC from patients. The method is a direct measurement of water content and therefore avoids the inaccurate and non-specific thickness to water content mappings used by current techniques.

THz medical imaging might be particularly suitable to measure water content in the cornea because its tissue

Corneal Morphology • 400 μm – 700 μm thick • ~ 79% water by volume/weight • Provides the majority of the focusing power of the eye • Refractive power and optical scattering strongly linked to water content

structure and geometry is generally much less varied than any other tissue in the body (e.g., in the lungs, liver, etc).

Furthermore, this variation is typically less than the wavelength of the target THz illumination enabling researchers to treat the cornea as a perfectly spherical, hydrated film sitting on top of a body of water (aqueous humour).

INITIAL TESTING Before they began developing their imaging tool, Dr Taylor and Dr Grundfest studied the cornea and its electromagnetic properties extensively.

From this, they proposed the first ever

Initial results were very promising but revealed unanticipated electromagnetic properties. Data from the millimetre wave measurements correlated well with CCT, while the THz and CCT measurements demonstrated no correlation. From their findings, the procedure appeared to alter the thickness of the cornea but not the CTWC. This perturbation in thickness, combined with the constant corneal water content, created a standing wave resolvable with the millimetre wave system and unresolvable with the THz system. This is the first experimental validation of a long-held belief that

Corneal disorders affect a considerable number of people worldwide, and can often result in chronic vision impairment – commonly requiring surgical intervention CTWC gradient models, as well as devised ways to identify CTWC from variations in THz reflectivity. The team initially tested their new imaging tool on the corneas of live rabbits. They attempted to measure their CTWC using a custom-built THz imaging system and millimetre-wave reflectometer and explored correlations between the imaging data and central corneal thickness (CCT) measurements.

trends in CCT are not necessarily associated with changes in CTWC. Two key concepts were extracted from this initial work. Firstly, the cornea is an optical thin film at THz frequencies.

Secondly, successful clinical translation of the technology will require noncontact measurements; a mode of operation generally not possible at THz frequencies.

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Above: Postdoctoral researcher Shijun Sung scanning his cornea with a prototype THz imaging system. The system scans the imaging beam while the patient’s head stays fixed using standard ophthalmic chin and forehead rests.

TRIAL AND ERROR: FURTHER TESTING To address the challenges identified in the first stage of work, Dr Taylor teamed up with researchers from TU Delft in the Netherlands, Chalmers University in Gothenburg, Sweden and VTT in Helsinki, Finland. A new optical system was designed, constructed and tested in partnership with colleagues specialising in physical optics. Pilot testing of the system was completed on a group of volunteers, which informed modification in preparation for upcoming pilot clinical trials. This work has produced what is believed to be the first non-contact THz image of a live human’s cornea ever to be published.

A PROMISING TECHNOLOGY Through their initial studies, Dr Taylor and Dr Grundfest collected a number of observations that will be useful for the research community and their development of THz frequency, ophthalmologic imaging tools. Their research has been met with a huge degree of excitement by the scientific community, with two of their research studies receiving the 2015 Terahertz Best Paper Award, in May last year.

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Subject 1: Left eye, en-face

Subject 1: Right eye, en-face

Reflectivity at 650GHz • Image acquisition time: ~20 seconds • Imaging subject stays completely stationary • Optics modification was crucial to imaging feasibility success Above: En face, non-contact, THz reflectivity maps of cornea from a volunteer subject. More reflectivity equates to higher water content.

Dr Taylor and Dr Grundfest’s imaging tool could make a real difference to the way corneal disorders are diagnosed and studied If perfected in a way that allows for accurate imaging and measurements of CTWC, Dr Taylor and Dr Grundfest’s imaging tool could make a real difference to the way corneal disorders are diagnosed and studied. The aim is to provide a non-invasive and accurate way of measuring hydration of the corneal tissue – something that is not yet possible. Not only does this research

offer a more efficient way of diagnosing corneal disorders, but also of exploring the relation between specific disorders and the tissue’s CTWC.

Behind the Bench Dr Grundfest

Dr Taylor

E: zdeis@seas.ucla.edu T: + 1 858 663 1823 W: http://www.ee.ucla.edu/zachary-taylor/ W: http://www.bioeng.ucla.edu/zachary-taylor-ph-d/ Research Objectives Dr Taylor and Dr Grundfest’s research focuses on developing a new imaging tool for non-contact, high resolution measurements of corneal hydration. Their collaboration since 2011 at The UCLA Henry Samueli School of Engineering and Applied Sciences has led to many publications and the Terahertz Best Paper Award in 2015.

Funding • National Institutes of Health (NIH) • National Science Foundation (NSF) • Telemedicine and Advanced Technology Research Center (TATRC)

Q&A

When and how did you first start developing the THz-based imaging tool to measure corneal hydration?

This imaging programme grew out of initial work on burn wound severity assessments using THz imaging.

Burn wounds are characterised by rapid accumulation and resorption of oedema (excess tissue water).

One of the key challenges with burn wound imaging is accounting for/ overcoming physiological variation, and we are always on the lookout for more predictable tissue systems. By chance, an ophthalmology colleague mentioned difficulties in obtaining accurate measurements of corneal hydration. A cursory review of corneal properties revealed extremely limited physiological variation and limited tissue heterogeneity on the relevant length scales. Our excitement was further increased by the inadequacy of optics based systems. We started with imaging excised bovine cornea which led eventually to NIH funding.

What do you feel have been your most promising findings so far?

Our most promising finding is that the cornea is a lossy thin film at THz frequencies. This means that corneal tissue may exhibit specific, narrow band properties as a function of morphology

Collaborators Sophie Deng, MD, PhD, Professor of Ophthalmology, Jules Stein Eye Institute, University of California at Los Angeles.

Bio Dr Grundfest received his BA from Swarthmore College in 1974, before completing his MD at Columbia University in 1980. He completed his Surgery Residency at the Cedars-Sinai Medical Center, and is currently a Professor at UCLA in the Departments of Bioengineering, Electrical Engineering and Surgery.

Dr Taylor received his BS in electrical engineering from UCLA in 2004. He later went on to study the same discipline at the MS and PhD level at UCSB. He currently works as an Adjunct Assistant Professor at UCLA in the Departments of Bioengineering, Electrical Engineering, and Surgery.

Contact Dr Zachary Taylor 420 Westwood Plaza 5121 Engineering V University of California Los Angeles, CA 90095-1600

and reflectivity measurements; combined with thickness maps this will enable accurate maps of absolute water content.

Optical thin film metrology is a mature field and we are currently adapting these techniques to the measurement of tissue water content.

sit perfectly still and the eye exhibits rapid movements, called saccades, that contribute to uncertainty in eye positioning. The timeline for clinical translation rests largely on our ability to overcome uncontrollable corneal movement.

Who do you believe could benefit the most from future applications of the tool developed through your research, and why?

We believe that ophthalmologists will benefit the most, initially. Numerous corneal diseases associated with abnormally raised CTWC are detected through visual assessment where the water content is sufficiently high to make the cornea appear cloudy. THz imaging may enable detection of these pathologies long before they become visually detectable thus enabling earlier intervention and improved patient outcomes.

What are your plans for future research and investigation?

We are focused on three major thrusts.

The first is extensive optical redesigns to enable rapid scanning. The second is incorporating phase sensitive source detector technology to enable rapid, coherent detection of magnitude and phase. The third is a large pilot human trial to understand the unique constraints associated with operating THz components in the ophthalmology clinic setting. Following successful completion of these engineering and evaluation tasks we plan to conduct a large scale clinical trial on patients undergoing corneal graft surgery.

In the long term, we would like to investigate applications in traumatic brain injury and are currently evaluating the physiological mechanisms that link elevated intracranial pressure to raised CTWC.

How long do you think it might take for your tool to enter healthcare settings and what remains to be done until then?

We are confident that our THz imaging technology can assess corneal water content in vivo. The major challenge for us will be uncontrollable patient movement.

Most of the planned use cases will be in the clinic with patients sitting in a chair, in a fashion similar to what you may have seen at the optometrist. People can’t

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Health & Medicine ︱ Prof Christian Beste

Unveiling the neurobiological processes behind cognitive control The biological underpinnings of human mental processes involved in acquiring knowledge, processing information and understanding experiences, have been substantially investigated throughout the years.

Prof Christian Beste, working at University Hospital Carl Gustav Carus, in Dresden, Germany, has carried out extensive research to try and understand the neural underpinnings of human goal-directed behaviour.

His studies are based both on clinical and empirical observations, as he believes an integration of both might lead to more exhaustive results.

T

he biological underpinnings of cognitive control, the mental function that allows individuals to multitask, perform goal-directed activities such as driving or preparing a meal, and develop strategies to cope when working on a number of things simultaneously, are not yet fully understood. Research in cognitive psychology and neuroscience has been thoroughly investigating the mental processes involved in cognitive control, trying to identify the parts of the brain and neurobiological processes associated with them.

COGNITIVE PSYCHOLOGY AND ITS LINKS TO BIOLOGY For many years, scientists have studied and tried to identify the mental mechanisms underlying human beings’ behaviour and actions.

In psychology, these are referred to as cognitive processes and include all mental processes involved in language use, attention, memory, perception, thinking, and problem solving. Cognitive psychology research is of particular importance as it aims to develop theories explaining the underpinnings of mental processes that occur in healthy individuals, as well as particular disturbances of these observed in those affected by particular medical or psychiatric disorders.

Once these theories are developed, neuroscientific studies can then try to link them to actual biological processes occurring in the brain, which can help to develop effective treatments for particular cognitive impairments or shed light on the biological dynamics behind them.

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Conceptual approach to study action control processes. Basic science approaches are combined with clinical/applied science approaches to examine the relevance of frontostriatal networks in action control.

Therapy evaluation biomarker

Basic science INTEGRATING RESEARCH AND CLINICAL WORK Throughout his career, Prof Beste has worked on investigating the neurobiological foundations of cognitive processes. As a student, he worked as an undergraduate research assistant in a Neurological department of the University Hospital (Ruhr-Universität Bochum) and at the Institute for Cognitive Neuroscience, Department of Biopsychology (RuhrUniversität Bochum). Prof Beste explains: “I did both (basic research and practical clinical work), because I felt that both ‘sides’ are important to integrate and that each of these sides can ‘learn’ so much from each other by simply trying to incorporate the ‘other perspective’.”

Fronto-striatal circuits

Experimental psychology EEG/MRT Neurobiology/genetics

Action control

Clinical/applied science and changes in structural neuroanatomy of the different disease,” says Beste.

Motivated to read more about the anatomy and neurophysiology of the ‘basal ganglia’, part of the brain that has been found to be associated with HD and PD, Prof Beste came across theories by Prof Peter Redgrave and colleagues, identifying the basal ganglia as playing a central role for those processes required to decide between different options or different ways to act. “I was really fascinated by this work, which stands very much

functions allowing human beings to work on different tasks simultaneously and complete goal-directed activities.

More recently, research aims to identify the parts of the brain and processes involved in cognitive control, particularly those necessary to cope in multi-tasking situations.

Prof Beste carried out a series of experiments that identified many biological mechanisms and anatomical structures involved in these mental processes, which were previously almost unknown: “We identified a number of ‘neurobiological bricks’ that are central for cognitive control mechanisms also playing a role in multi-tasking,” he says. “Moreover, we were able to show which functional neuroanatomical structures are causally involved in multitasking and determine the efficacy and strategy applied to cope with multitasking situations.”

We identified a number of ‘neurobiological bricks’ that are central for cognitive control mechanisms

At the time, his work focused on the neuropsychological diagnostics of neurodegenerative diseases, such as Parkinson’s disease (PD) and Huntington Disease (HD), progressive diseases of the nervous system characterised by a variety of symptoms, including jerky body movements and cognitive impairments. “I became frustrated by the very apparent oversight that diagnostic procedures were not orientated and constrained by the underlying pathoneurophysiology

in contrast to ‘research traditions’, that focus on the importance of the neocortex in these basic cognitive functions,” says Prof Beste. Inspired by this research, Prof Beste became increasingly interested in the brain mechanisms that enable ‘action selection’ – the essential ability that allows individuals to select actions to perform in the moment.

THE BIOLOGICAL PROCESSES BEHIND COGNITIVE CONTROL Most cognitive psychologists define cognitive control as a set of mental

The professor and his colleagues found that levels of GABA and glutamate in a part of the basal ganglia called the striatum could predict particular neurophysiological processes measured using electroencephalography (EEG), a method of recording electrical activity in the brain. They also found

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Illustration of functional neuroanatomical networks and neurobiological systems involved in multi-component behaviour and multi-tasking.

that diseases specifically affecting microstructural elements of the basal ganglia had profound effects on cortical neurophysiological mechanisms underlying cognitive control.

LINKS TO NEUROLOGICAL DISEASES AND PSYCHIATRIC DISORDERS In a further series of experiments, Prof Beste and his colleagues evaluated the role of EEG measures and experimental paradigms related to cognitive control as biomarkers for the progression of neurodegenerative diseases as well as neuropsychiatric disorders. They found that combining neurophysiological measures with experimental paradigms tapping into the functions of some of the striatum’s circuits helped to point out early disease progression in PD and HD and are useful to discover novel facets of deficits in neuropsychiatric disorders, e.g., attention deficit hyperactivity disorder (ADHD) or Tourette Syndrome. The changes tracked could predict clinically relevant parameters for progression of these diseases better than parameters that are currently in place. The same measures also appeared able to effectively assess subtle changes in medication treatments in these diseases and disorders.

Multitasking and functions necessary for this are very central for our ability to cope with requirements in daily life

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A COMPREHENSIVE APPROACH Prof Beste’s research provided great insight into the neurobiological dynamics of mental processes associated with goal-directed activities.

Through an integrative approach that combines basic and clinical research, he and his colleagues were then able to identify particular areas and processes in the basal ganglia which are associated with multi-tasking and appear to be adversely affected in neurodegenerative diseases and psychiatric disorders.

These brain areas and processes could also be impaired in other conditions that are characterised by impairments in cognitive control, including psychiatric disorders. Excitingly, his findings may well provide a basis for further research, ultimately leading to an increasingly accurate understanding of the biological underpinnings behind goal-directed behaviour in humans.

Behind the Bench Professor Christian Beste

E: christian.beste@uniklinikum-dresden.de T: +49 351 458 7072 W: http://www.actionlab.de W: http://www.researchgate.net/profile/Christian_Beste

Research Objectives Prof Beste is a neurophysiologist and cognitive neuroscientist who aims to combine clinical and basic research to understand the neural underpinnings of human goal-directed behaviour and neurology cognitive control.

Funding • Deutsche Forschungsgemeinschaft (DFG) • Bundesministerium für Bildung und Forschung (BMBF) • Else-Kröner Fresenius Stiftung (EKFS)

Q&A

You have taken a novel approach to your research into neurodegenerative disorders by combining clinical and basic research. How did you come to study this field?

When I was a student I was working in many labs on different topics related to cognitive neuroscience, but I also worked in a neuropsychological department at a hospital. There, I was involved in neuropsychological testing and diagnostic procedures in patients with various neurological diseases. Doing so, I wondered increasingly whether the tests we apply in clinical routines do really have a solid neurobiological basis. I thought that if this is not the case, then we are very likely to miss diagnostic sensitivity and specificity. At this time, I mainly worked with patients with neurobiological basal ganglia diseases (e.g., Parkinson’s Disease, Huntington disease). This led me to read about the neurobiology and physiology of

• Friede Springer Stiftung • BIOGEN • GENZYME Bio After graduating in Psychology in 2006 at the Ruhr-Universität Bochum, Prof Beste received his PhD in 2007. Before establishing an independent EmmyNoether Group at the Ruhr-Universität Bochum in 2012, he was Post-Doc at the University of Münster, TU Dortmund and the MRC Cognition and Brain Science Unit (Cambridge).

basal ganglia-prefrontal cortical networks in relation to cognitive functions and I became fascinated by this topic. My previous, current and future research is a constant development of aspects that had fascinated me when I was an undergraduate student.

Why is multi-tasking so vital for cognitive function?

I think that multitasking – or what we think of as multitasking – is very central for our ability to cope with requirements in daily life. I cannot think of a situation in which such abilities may not be relevant – except deep meditation for example. Because ‘multitasking’ is a complex cognitive function and requires large-scale network organisation, these processes especially are very central when thinking of diagnostic procedures suitable to detect even subtle changes in brain functions.

What most fascinates you about neurobiological research?

It is the complexity of interacting neural processes and how this complexity may

Contact Professor Christian Beste University Hospital Carl Gustav Carus Dresden and Institute of Psychology TU Dresden Fetscherstraße 74 01307 Dresden Germany

result in seemingly simple observable behaviour that fascinates me.

How can neurodegenerative diseases help us investigate brain function?

It is not so much the neurodegenerative disease per se, but various diseases that affect brain function – including psychiatric disorders. I strongly believe that when you are interested in the question “How does the brain manage to deal with situations requiring cognitive control e.g. ‘multitasking situations’?” a good strategy is to approach this question from different angles. Disorders of the CNS provide these different angles necessary to gain a more comprehensive picture about the underlying neurobiological mechanisms. In particular, when there are clear and specific neurobiological changes (e.g., in the case of XDP patients) this approach allows us to gain insights into the mechanisms important for human goal-directed behaviour.

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Health & Medicine ︱ Dr Marion Kavanaugh-Lynch

Community collaborations targeting breast cancer Dr Marion Kavanaugh-Lynch directs the California Breast Cancer Research Program of the University of California. Her latest work with the QuickStart programme in California offers coaching for partnership teams formed between communities and academics to conduct participatory research. With a focus on the environmental factors in breast cancer and social disparities in breast cancer, she has forged an important partnership between the California Breast Cancer Research Program, Commonweal and the Orange County Asian and Pacific Islander Community Alliance.

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reast cancer affects millions of people around the world and innovative research is essential to end this disease. The state of California alone invests $15 million into various research projects each year, raising the important question – what is the best way to invest these precious resources?

At the California Breast Cancer Research Program, this responsibility falls to its director – Dr Marion Kavanaugh-Lynch.

Dr Kavanaugh-Lynch and partners from Commonweal and the Orange County Asian and Pacific Islander Community Alliance are developing a programme to encourage members of the local community to engage in their own research projects. Approximately 70% of breast cancer cases could have an environmental cause and, with wide

ranging social disparities in outcomes, the more insight that communities can contribute, the better.

QUICKSTART This innovative programme, known as QuickStart, delivers face-to-face, telephone-based, and online sessions for either established or newly formed partnerships. Throughout the programme, teams develop a research plan. This includes firstly, identifying the problem and formulating a research question; secondly, designing a research plan; and thirdly, developing a budget and writing a grant proposal.

All partnerships include a Community Co-Principal Investigator and an Academic Co-Principal Investigator.

The Co-Principal Investigators share

QuickStart Faculty provide support to Fellows

leadership on the research project and ensure an adequate representation of both community and academic perspectives. The Community CoPrincipal Investigator represents a group within the community and should have the trust of their organisation, as well as the skills to communicate with all members of the research team and the broader community, who may be impacted by the research. The Academic Co-Principal Investigator must have allocated time for research. Their skill set and knowledge base should align as closely as possible to the research interests of the Community Co-Principal Investigator. The QuickStart programme allows for thorough development of the research question and helps partnerships discern where specific expertise is required.

QuickStart programme are made evident to applicants in the brochures and application forms they receive. For those willing to undertake the commitment, including four days of face-to-face sessions, online weekly assignments, and telephone coaching, the benefits are huge. The three-month programme provides opportunities for both the academics and the community members involved.

Academics can gain critical insights into the realities of breast cancer in specific communities, as well as improve their likelihood of receiving funding for a project, due to the demonstrably high level of stakeholder interest.

Likewise, community members can gain valuable research skills – improving the reputation of the organisation they represent and gathering data for their own campaigns. Both academics and communities increase their chance of creating actual long-term change to policy and practices.

COLLABORATIONS AND PARTNERSHIPS The type of research created by partnerships in the QuickStart programme is community-based participatory research. The programme specifically focuses on the environmental causes of breast cancer and social disparities experienced by breast cancer sufferers.

Approximately 70% of breast cancer cases could have an environmental cause

Eligibility is largely unrestricted, provided participants are willing to commit to all aspects of the programme.

The practicalities of taking part in the

They will also maximise the potential dissemination of their research, having actively engaged with the groups most likely to benefit during the primary stages.

It is hoped that successful completion and evaluation of this project will lead to an established method of delivering a curriculum of this nature to the public. Ultimately, it is expected that the partnerships that participated in the QuickStart

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Capacity building for Community and Academic Fellows

programme will be able to submit grant applications to the California Breast Cancer Research Program Community Research Collaborations awards, as well as to other funding sources.

In order to maximise the reach of the project, Dr Kavanaugh-Lynch has formed important partnerships with organisations to enable expansion of the opportunity to diverse cross sections of Californian communities.

These partner organisations include Commonweal and the Orange County Asian and Pacific Islander Community Alliance (OCAPICA). Commonweal is a California-based non-profit health and environmental research institute which seeks to engage in activities to improve public health. Their activities span education, research and charitable work, with the impact of their programmes not only improving the health of individuals, but also enhancing the health of the global environment.

OCAPICA works to benefit the health and wellbeing of Asian and Pacific Islanders living in Orange County, California. Their involvement

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QuickStart has been described as “invaluable and career changing” and “an excellent combination of scientific content and team building exercises” in this project is testament to the importance of encouraging maximum diversity of groups benefitting from and contributing to the QuickStart programme. The project is supported by the National Cancer Institute of the National Institutes of Health.

Previous programmes have resulted in successful research funding for more than half of their participants, and the range of topics studied has been diverse. Projects can be broad in their intended impact or can target a small and specific group of people. Examples of previous projects include an investigation into the effect of benzene exposure on air pollution, as well as a study encouraging teenage mothers to persist with breastfeeding to protect against future incidences of breast

cancer. QuickStart has been described as “invaluable and career changing” and “an excellent combination of scientific content and team building exercises”.

The academic accomplishments of Dr Kavanaugh-Lynch within the field of breast cancer research, alongside her continued advocacy for the voices of all members within a community to be heard, will ensure the QuickStart programme helps all participants to achieve their full potential.

More info on QuickStart: http://cbcrp.org/funding-opportunities/ crc/quick-start-training.html www.commonweal.org/program/ quickstart

Behind the Bench Dr Marion Kavanaugh-Lynch E: marion.kavanaugh-lynch@ucop.edu T: +1 510 987 9878 W: http://cbcrp.org/index.html

Research Objectives QuickStart supports communityacademic teams in conducting community based participatory research (CBPR) related to breast cancer. This is a rare opportunity to get technical assistance directly from a potential funder on a research proposal that the teams develop over the course of the programme. Participants build their partnerships and workshop a research idea through in-person and online sessions. QuickStart participants develop the expertise to collaboratively develop research questions, plans and grant proposals.

Funding National Institutes of Health (NIH)

Q&A

When developing the programme did you need to build in flexibility to accommodate the different cultures throughout California?

Yes. We continually evaluate our progress and refine aspects of the programme so that community and academic organisations across California can participate. For example, we have shortened the schedule so that partnerships attend four days of in-person sessions (down from six).

Also, we have revised our application process to provide technical support to organisations that are interested in participating but would like our support in finding a partner for the programme.

Collaborators • Senaida Poole, PhD, Program Officer, Community Initiatives and Public Health Sciences, California Breast Cancer Research Program (CBCRP) • Heather Sarantis, MS, Women’s Health Program Director, Commonweal • Mary Anne Foo, MPH, Executive Director, Orange County Asian and Pacific Islander Community Alliance (OCAPICA) Bio Dr Kavanaugh-Lynch’s accomplishments include championing the role of advocates and survivors in the peer review process, developing a successful model for funding community-based participatory

Can this method of programme be applied nationwide or even worldwide?

Absolutely. We are exploring different methods of delivering the curriculum to eventually deliver a programme that can be delivered to anyone, anywhere.

What do you hope the impact of involving more people from the community in research projects will be?

In order to promote global well-being and health, scientific discoveries must be applied to address the lived realities of people in their communities. Academic scientists cannot do this alone, and neither can community organisations.

But working together, they can transform their worlds.

research and developing rigorous evaluations of the programme. In recent years, she led a national panel that developed research strategies to explore the role of environmental contaminants in breast cancer, breast cancer disparities, and prevention of breast cancer.

Contact Marion H E Kavanaugh-Lynch, MD, MPH California Breast Cancer Research Program University of California, Office of the President, 300 Lakeside Drive, 6th Floor, Oakland, CA 94612-3550 USA

What has the feedback from the academic Co-Principal Investigators been like? Have they had any surprises along the way?

They have learned that forging productive partnerships and working collaboratively can be very timeconsuming. But they have found that the rewards are profound. Their research theories come alive and transform lives.

Is there scope to tackle research questions about other diseases using a programme like QuickStart?

This sort of research partnership can be used to tackle any disease that the community is concerned about.

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Health & Medicine ︱ Professor Christian Schulz

Dual origins of tissue macrophages Macrophages – large white blood cells that play a role in tissue homeostasis and immunity – have long been thought to derive solely from monocyte cells in the circulating blood. Accumulating evidence now shows that a large proportion of the macrophage populations ‘resident’ within tissues is in fact derived during embryonic development, independent of monocytes. Professor Christian Schulz from Ludwig-Maximilians-Universitat, Munich, is at the forefront of this research. He hopes that understanding the molecular regulation of these cells during development and within their tissue of residence will help design interventions to treat a wide spectrum of diseases in which they have been implicated.

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M

ost people with an interest in biology will be able to tell you that macrophage cells are an important component of our immune system. By engulfing and digesting microbes, cellular debris, foreign substances and anything else that isn’t a healthy body cell, they help to protect us against injury and infection. It is now clear that macrophages are present in all tissues under steady state conditions, and that these tissue-resident cells play tissue-specific roles: tissue remodelling, angiogenesis (blood vessel formation) and regulation of cell metabolism, to name a few. Tissue-resident macrophages are exquisitely adapted to their local environment, acting as regulatory cells of tissue function, and this makes them an attractive target for modern medicine.

Understanding the origin and regulation of these cells will be an important first step in the development of intervention strategies to control and instruct macrophage populations.

TWO BEGINNINGS During development, haematopoiesis (including macrophage production) first takes place in the yolk sac (yolk sac haematopoiesis). Soon thereafter the foetal liver becomes a hematopoietic site expanding hematopoietic stem cells that migrated from the aortogonado-mesonephros region (foetal haematopoiesis) but also of macrophage progenitors that derived from the yolk sac. Towards the end of gestation, foetal haematopoiesis switches from the liver to the bone marrow.

Until recently, it was believed that tissueresident macrophages were constantly replenished from pools of circulating monocytes (macrophage precursors) produced by foetal haematopoiesis.

However, this concept has been questioned following reports that macrophages in the brain, liver, skin, and many other organs proliferate and self-renew. For example, donor-derived

Fate mapping (green) of yolk-sacderived alveolar macrophages

tissue macrophages have been reported to persist for many years in transplanted liver and skin in human patients. The hypothesis was therefore raised of two distinct origins of macrophages: the first being yolk sac haematopoiesis which produces mature macrophages, the second being foetal haematopoiesis which produces blood monocytes.

Professor Christian Schulz and colleagues therefore decided to reinvestigate the origin and development of tissueresident macrophages.

TWO POPULATIONS Among various biological tools, the team used a technique known as pulselabelling to map the fate of developing macrophage cells in mouse embryos.

They showed that macrophages derived from the yolk sac (YS) circulate in the blood and colonise the embryo, starting with the head region, nine to ten days after fertilisation. By ten days after fertilisation, the cells could be detected in most tissues, where they continued to divide. They remained detectable in tissues throughout development.

From twelve days onwards (around the time that foetal haematopoiesis begins

The team used a technique known as pulselabelling to map the fate of developing macrophage cells in mouse embryos

in the liver) a second population of macrophages appeared.

Prof Schulz and colleagues set out to determine the origin of these newcomers. They re-examined macrophage development, this time using mice with a deleted Myb gene.

The team had previously used this mouse model to show that the protein Myb is required for foetal but not yolk sac haematopoiesis. The results were as hypothesised: YS-derived macrophages developed as normal but the second population of macrophages did not appear, indicating that they are indeed of the foetal haematopoiesis. YS-derived macrophages accounted for most macrophages in tissues of embryos at 16 days after fertilisation.

Importantly, fate mapping models provided evidence that YS-derived macrophages persist in many tissues of adult mice, in some of them (i.e., brain, liver) throughout life. These findings support the view that there are two origins of macrophages in adult tissues; the first are derived from the yolk sac during early embryonic development and the second are derived from foetal haematopoiesis in the liver and later the bone marrow.

A CLEARER VISION Thanks to this work and further work

by Prof Schulz and others, we now

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First appearance of macrophages in the mouse embryo at embryonic day E9.75 (before the initiation of definitive hematopoiesis)

Above: Yolk sac-derived macrophages develop in tissues in the absence of foetal definitive hematopoiesis

Right: Appearance of macrophages in the mouse embryo 9.75 days after fertilisation (before the initiation of foetal definitive hematopoiesis)

have a clearer vision of the origin and development of macrophages.

Macrophage progenitor cells appear in the yolk sac and then invade the developing embryo. They colonise all foetal tissues, particularly in the head where they give rise to all of the brain microglia, which will continue to selfrenew throughout adulthood.

Later, foetal haematopoiesis produces monocytes. They circulate in blood and need to be continuously renewed by definitive haematopoiesis. In some organs, they differentiate and replace tissue macrophages over time (e.g., lung). However, this population seems more important in the setting of tissue injury or inflammation. Here, monocytes are recruited from blood into tissues, where they differentiate into phagocytes to fulfil specific jobs/duties. They may also renew resident macrophages if needed.

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E9.75

Yolk sac

Macrophages

Embryo 500 Âľm

Thanks to this work... we now have a clearer vision of the origin and development of macrophages MOVING ON Professor Schulz has now turned his attention to the biology of macrophages in relation to their developmental paths in cardiovascular disease. Similar to the work described in this article, current projects utilise in vivo lineage tracing and fate mapping, as well as models for time- and site-specific gene deletion.

The team also carries out genomic analysis and proteomics to determine macrophage programming by the local environment and disease-specific signals. This work could eventually aid the development of strategies to control the remodelling of cardiovascular tissues, for example, after myocardial infarction.

Behind the Bench Prof Christian Schulz

E: christian.schulz@med.uni-muenchen.de T: +49 89 4400 73092 W: http://www.sfb914.med.unimuenchen.de/principal_investigators/principal_investigators/schulz_christian/index.html W: http://www.klinikum.uni-muenchen.de/Medizinische-Klinik-und-Poliklinik-I/de/Research/BasicResearch/People/index.html

Research Objectives Prof Schulz’s work focuses on the role of macrophages in tissue homoeostasis and inflammation, specifically processes associated with cardiovascular disease.

Funding • The German Research Foundation (DFG), specifically the Collaborative Research Centre (SFB) 914 (project A10) • The German Center for Cardiovascular Research (DZHK) • Leopoldina National Academy of Sciences

Q&A

What first piqued your interest in tissue-resident macrophages?

I was intrigued by previous reports that in organ transplant patients tissue macrophages of donor origin persisted for years. This indicated their self-renewal capacity and their independence of bone marrow haematopoiesis.

Can you give some examples of the tissue-specific functions of tissueresident macrophages?

• Initiation and resolution of inflammationx • Clearance/waste disposal (liver, spleen) • Regulation of metabolism • Angiogenesis and vascular remodelling • Development: bone morphogenesis, ductal branching, neuronal connectivity A fate mapping model was used to report the persistence of yolk-sac-

Collaborators • Jon Frampton (University of Birmingham) • Frederic Geissmann (Memorial Sloan Kettering Cancer Center New York) • Elisa Gomez Perdiguero (Institute Pasteur Paris) • Jeff Molkentin (Cincinnati Children’s Hospital Medical Center) BIO Dr Christian Schulz is Professor of Cardiovascular Immunology at the LMU Munich. He trained in Internal Medicine and worked as a Postdoctoral Fellow at the Technical University

derived macrophages in adult tissueresident macrophage populations. Can you explain briefly how this worked?

We used a so-called inducible Cre/lox system, which enabled us to generate cell type- and time-specific expression of a fluorescent protein. By inducing expression specifically during early yolk sac haematopoiesis, we were able to identify these fluorescent YS-derived cells later in embryonic development as well as in adult animals.

The development of tissue-resident macrophages has been characterised using mouse models. Are there likely to be differences between macrophage development in mice and humans?

A yolk sac also exists in humans.

Munich. He then moved to the United Kingdom for postdoctoral work at the Centre for Molecular & Cellular Biology of Inflammation and later became a Senior Lecturer at King’s College in London.

Contact Prof Christian Schulz Medizinische Klinik und Poliklinik I Klinikum der Universität München Ludwig-Maximilians-Universität München Campus Großhadern Marchioninistraße 15 81377 München, Germany

It produces haematopoietic cells, which are thought to enter the foetus in early life. However, whether this concept identified in mice (and zebrafish) holds true in humans is unknown. Notably, tissue macrophages in humans can persist autonomously for years [see comment above on transplant patients].

What are the potential therapeutic targets of your research?

We need to understand the regulation and programming of macrophages that fulfil tissue-specific functions in humans. This could potentially be used as therapeutic targets in chronic tissue inflammation to induce inflammation resolution and improve metabolism and remodelling.

I was intrigued by previous reports that in organ transplant patients tissue macrophages of donor origin persisted for years

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Health & Medicine ︹ Professor Ghada Bourjeily

The dangers of sleep disordered breathing in pregnancy Professor Ghada Bourjeily and her team from Brown University are focusing their research efforts on understanding the manifestation of sleep disordered breathing (SDB) in pregnancy and the consequent impact on maternal and foetal health.

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e spend one third of our life sleeping. As we rest, damaged tissues are repaired and cognitive function and energy levels are restored in preparation for an active day ahead. Sleep deficiency can greatly impact our health, highlighting the importance of good quality sleep.

Abnormal breathing patterns during sleep, otherwise known as Sleep Disordered Breathing (SDB), are a common cause of sleep inadequacy. SDB includes a wide spectrum of conditions, from snoring to obstructive sleep apnoea (OSA). OSA occurs when throat muscles relax during sleep, causing the upper airway to collapse. Obstruction reduces airflow for around ten seconds or more, resulting in low levels of blood oxygen saturation (hypoxia). In the long-term, OSA-associated hypoxia can lead to potentially fatal conditions such as

hypertension, cardiovascular disease and diabetes mellitus.

PREGNANCY-ASSOCIATED SDB Interestingly, pregnant women are at a higher risk of developing SDB, due to the physiological changes that occur during pregnancy. For example, plasma volume increases and capillary engorgement causes airway mucosa to thicken, resulting in nasal congestion as the lining of the nose, larynx and trachea swells. This can lead to gestational rhinitis development, which usually improves immediately after delivery.

Nasal congestion is a major risk factor for SDB, therefore pregnant women are particularly at risk. Furthermore, during the later stages of pregnancy, the gravid uterus causes the diaphragm to elevate, reducing the functional residual capacity of the lungs by 20%. This may impact

Sleep disordered breathing research team. Back row, left to right: Greg Salgueiro, MS, RD, LDN, CIC; Rebecca Lynn; Susan Martin, LDN, IBCLC; Cindy Brosnan, RRT; Patrizia Curran, MD; Christine Allenson, MA, OT, CHES Front row, left to right: Beth Hott, BA; Tamara Sequeira, RN; Maggie Bublitz, PhD, Ghada Bourjeily, MD, Annaly Aldana, BA, Eva Adodoadji, MD, MPH

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oxygen reserve and further contribute to upper airway collapsibility. Snoring is also much more frequent in pregnant (14–45%) compared to non-pregnant women of reproductive age (4%).

The strong association between SDB and pregnancy raises a number of questions: i) does SDB cause detrimental effects in the mother and foetus / newborn? ii) what physiological or biological factors predict the development of SDB in this young population? and iii) what are the mechanisms that result in these adverse effects? Professor Bourjeily and her colleagues have dedicated their research efforts to investigating these interesting questions.

GESTATIONAL HYPERTENSIVE DISORDERS Professor Bourjeily and others have shown that pregnancy-associated SDB (including snoring and OSA) increases the risk of developing gestational hypertensive disorders such as hypertension and pre-eclampsia, even after considering other risk factors such as obesity. Gestational hypertension complicates approximately 6% of all pregnancies and pre-eclampsia is a severe pregnancy disorder, characterised by high blood pressure and either a high level of protein in the urine or other systemic manifestations. The condition may deteriorate further to ‘eclampsia’ and seizures may occur which may threaten the lives of the mother and child.

Pre-eclampsia is also associated with pulmonary oedema, liver abnormalities and renal failure. Though pre-eclampsia is a short-lived condition, it has been suggested to be a precursor of future development of cardiovascular disease.

So, how does SDB cause pre-eclampsia?

One of the possible mechanisms could be placental hypoxia. In a study, performed by Professor Bourjeily and her team, a quantitative analysis of immunohistochemical markers of hypoxia were compared between the placentas of pregnant women with OSA/snoring and non-snoring controls.

Expression of the hypoxia marker carbonic anhydrase was more prevalent in OSA placentas (91.3%) compared

to the controls (57.5%). In addition, the team has demonstrated that placenta secreted blood markers are altered in women with OSA compared to controls.

Professor Bourjeily hypothesises that placental hypoxia that may occur as a consequence of SDB could trigger a cascade of events that lead to preeclampsia. The hypoxic placenta secretes a variety of different soluble molecules into the bloodstream that impacts maternal endothelial function.

In fact, endothelial abnormalities have been demonstrated in patients with OSA, regardless of the severity level.

Endothelial dysfunction is characterised by an imbalance of hormones that control blood pressure. Consequently, hypertension can develop which, if not monitored, can lead to heart disease.

Furthermore, inflammation could be the missing link between SDB

and hypertensive disorders. Sleep disturbance is associated with high levels of interleukin-6 in the later stages of pregnancy. This inflammation marker is strongly associated with preeclampsia. Nevertheless, much more research is needed to fully understand this relationship.

GESTATIONAL DIABETES In a study conducted on 1000 patients, Professor Bourjeily also demonstrated that there is a significant association between gestational diabetes and third trimester SDB (which includes snoring, gasping and apnoeas) regardless of other factors such as BMI and smoking.

In addition, in a large populationbased sample that included over 1.5 million women, a diagnosis of OSA was associated with an increased risk for gestational diabetes, after adjusting for multiple risk factors.

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SDB in PREGNANCY

AIRFLOW LIMITATION

SYMPATHETIC ACTIVATION

OXIDATIVE STRESS

AIRWAY COLLAPSIBILITY WEIGHT GAIN AIRWAY EDEMA FLUID SHIFTS HORMONES

INFLAMMATION

Gestational diabetes affects 2–10% of pregnancies and is characterised by glucose intolerance. Serious pregnancy complications can result from gestational diabetes, including pre-eclampsia, increased risk of caesarean delivery and preterm labour. In addition, gestational diabetes is strongly associated with future development of type II diabetes, a highly morbid condition.

Excessively high blood glucose levels, as a result of SDB, may be related to a range of interlinking factors. For example, airflow limitation and hypoxia can result in increased sympathetic activation (part of the nervous system

AROUSALS

INFLAMMATION

OBESITY

INTERMITTANT HYPOXEMIA

THROMBOSIS

ENDOTHELIAL DYSFUNCTION

PLACENTAL INJURY AND DYSFUNCTION

PRE ECLAMPSIA Proposed mechanism for the link between sleep disordered breathing and preeclampsia

Dr Bourjeily is currently investigating additional pathways that may link SDB and gestational diabetes.

FOETAL RISK AND DELIVERY Results from studies focusing on the consequences of SDB on foetal outcomes are conflicting. SDB (both snoring and OSA) appears to be associated with preterm birth (birth occurring before 37 weeks gestation) in multiple studies, including Dr Bourjeily’s.

The risk of growth restriction and small for gestational age remains controversial.

Dr Bourjeily is currently investigating the risk of growth restriction and other neonatal outcomes in a large population-

FUTURE STUDIES The work of Professor Bourjeily and her colleagues has greatly advanced our knowledge regarding the effects of pregnancy-induced SDB. However, more research is needed to fully understand mechanisms behind the association. The team are now focussing their research efforts on exploring why SDB prevalence more than doubles in later stages of pregnancy compared to earlier stages.

By determining the physiological factors that predict the onset of SDB in late pregnancy, the team are aiming to create a model, alongside simple testing, that can be used as a preventative tool, to identify SDB development. The patient

Professor Bourjeily and others have shown that pregnancyassociated SDB (including snoring and OSA) increases the risk of developing gestational hypertensive disorders such as hypertension and pre-eclampsia involved in the ‘fight or flight response‘), which can i) inhibit insulin secretion from the pancreas, ii) exacerbate insulin resistance (cells cannot respond to insulin signals which trigger glucose uptake from the blood) or iii) stimulate glucose release from liver cells into the bloodstream. Additionally, oxidative stress can damage pancreatic beta cells, where insulin is produced, resulting in a reduction in insulin secretion.

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based dataset that linked maternal and neonatal records. Other potential adverse foetal impacts include reduced heart and growth rate. Despite these inconsistent findings, the majority of studies provide supportive evidence that suggests that women who snore, have short sleep duration and OSA are at a higher risk of caesarean delivery, compared to controls.

can then be treated before the SDB becomes more severe. As SDB is a relatively easy condition to treat with ways that are not thought to negatively impact the foetus, investigating this condition that may impact up to a third of all pregnant women should be a priority.

Behind the Bench Professor Ghada Bourjeily

E: hada_bourjeily@brown.edu T: +1 401 444 8664 W: http://vivo.brown.edu/display/gbourjei W: http://bit.ly/2y61vTO

Research Objectives Prof Bourjeily’s work studies the links between sleep disordered breathing, pregnancy and perinatal outcomes. She looks at both the effect on pregnancy outcomes and the potential for long-term implications for these women.

Funding NIH, Chest Foundation Collaborators • Geralyn Messerlian, PhD Professor of Pathology, Brown University • Maggie Bublitz, PhD, Assistant Professor of Psychiatry and Human Behavior and Medicine, Brown University

Q&A

Why is sleep disordered breathing so prevalent in pregnant women?

SDB is more prevalent in pregnant women compared to non-pregnant young women likely because so many physiological changes occur in pregnancy that may impact the upper airway, the lungs and the heart. The upper airway, including the nose and the laryngeal area, appears to be more oedematous (swollen) in pregnancy and this may significantly contribute. Resting lung volumes also change in pregnancy, potentially impacting the upper airway. Pregnancy hormones may also play a role in the development or the worsening of this condition in pregnancy.

We do not understand yet which of these changes has the biggest impact on its development but are hoping to get more answers in the near future.

SDB is related to gestational diabetes and hypertensive disorders. Are there any other conditions that could actually be caused by SDB? Though it makes biological sense that gestational hypertension and gestational diabetes could be caused by sleep disordered breathing, research to date does not confirm causality.

Hence, we talk about associations.

• Fusun Gundogan, MD, Associate Professor of Pathology, Brown University • The obstetric medicine team at Lifespan under the directorship of Dr Lucia Larson, Associate Professor of Medicine, Brown University, division director of Obstetric Medicine, and Peg Miller, Associate Professor of Medicine, Brown University and Chief of Women’s Services at Lifespan • Marshall Carpenter, MD, Maternal Fetal Medicine, and the obstetric team of providers who help facilitate patient recruitment Bio Ghada Bourjeily is associate professor of medicine at the Warren Alpert Medical

Future studies that look at the impact of therapy on these outcomes or establishing a temporal relationship between these conditions can help us establish causality further. In a recent study we have shown that SDB has been associated with other severe maternal illness such as pulmonary oedema, congestive heart failure, risk of admission to the intensive care unit, as well as an increased risk of a hysterectomy.

There also appears to be an elevated risk of requiring a Caesarean delivery once diagnosed with SDB. How does pregnancy-induced SDB affect foetal health?

The foetus and the placenta may be target organs in the case of SDB, just like the heart and the kidney may be impacted by SDB. To date, there has been quite a few data linking SDB to preterm birth with a recent study showing an increased risk of both induced and spontaneous preterm birth.

Data are inconsistent in showing evidence of growth restriction. What is lacking in the literature is an understanding of the timing of development of SDB and how SDB pre-dating pregnancy would differently impact foetal growth. For instance, is it possible that long standing SDB may cause some protective mechanisms to take effect further protecting the foetus compared to SDB that develops in pregnancy? These are questions we do not have answers to at this time.

School of Brown University in the divisions of pulmonary, critical care and sleep medicine, and obstetric medicine. She is the director of women’s research at the women’s medicine collaborative at the Miriam Hospital and director of the pulmonary disease in pregnancy program.

Contact Ghada Bourjeily Associate Professor of Medicine 146 West River Street, Providence RI 02904 USA

Are women affected by the impacts of pregnancy-induced SDB, following birth?

This is an area where the research is still lacking. In an indirect way, we could say that if SDB is increasing the risk of pre-eclampsia for instance, and pre-eclampsia is associated with short term risk factors and long term adverse cardiovascular risk factors, SDB may be increasing the risk of future cardiovascular disease. However, we (the collective we) have not yet studied how or whether SDB could impact (mediate or catalyse) the link between preeclampsia and cardiovascular disease, or the link between gestational diabetes and type II diabetes for instance.

What further research will you be conducting?

We would like to focus our future research on better understanding mechanisms of these associations as those may be quite different in the pregnant population due to the duration of the exposure, the accelerated outcomes, the hormonal milieu and other factors associated with pregnancy.

We also would like to identify therapeutic targets and understand barriers to therapy in women who may be minimally symptomatic.

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Thought Leader

HRA: making science open, managed and well-funded Back in 2015, the Health Research Alliance (HRA), a multi-national consortium of non-profit organisations in the field of biomedical research, announced the selection of Dr Maryrose Franko as its new Executive Director. Bringing over 70 nonprofit and non-governmental funders of biomedical research together, the HRA is the number one place to go for non-profit organisations seeking to enhance return on their investment in biomedical research. But HRA is about much more than just the “nuts and bolts” of grantmaking. Dr Franko leads the organisation in its commitment to popularising research and taking care of science’s most valuable asset – its people.

H

ealth Research Alliance’s story began in 1998, when a group of private funders met to discuss the role of philanthropy in financing medical research in America. Some of them later began to meet regularly, which led to the foundation of Clinical Research Alliance. CRA efforts were aimed mainly at gathering data on medical research and sharing knowledge, best practices and experience among its members.

But, when the CRA convened over 70 representatives of biomedical research foundations and voluntary health agencies in 2004, they broadened the scope of their activities to include issues that did

learning in areas like grantmaking and communicating impact. I also do that by making sure HRA gives its members a voice in conversations important to HRA members, thus increasing the openness and reusability of the science we fund – not only publications, but also data and other research outputs.

I work closely with the co-chairs of each HRA’s Interest Groups to set agendas, convene meetings/webinars, facilitate communication with members of the Groups and conduct research as needed to guide actions. This means I spend quite a bit of time building relationships

model for scholarly communication which is why I was honoured to be invited to serve on the COS board.

Can you tell us about the HRA and briefly outline HRA’s core mission?

In 2005, the HRA was formally incorporated. 21 funders formally joined the Alliance in 2006. Today, we now have almost 80 members, and are still growing!

Our mission and core values are all about maximising the impact of biomedical research to improve human health.

To achieve that we collect and share data and analysis about non-profit funding

Our mission and core values are all about maximising the impact of biomedical research to improve human health not fall neatly into the clinical research category. Thus, in 2005, CRA became the HRA which since then, has built one of the largest searchable databases (called the HRA Reporter) of grant awards covering over ten billion dollars’ worth of available grant-money from 2006 till today, supported young scientists at the early stage of their careers, and advocated for open access to scientific data and publications. Now, with Dr Maryrose Franko, at the helm, HRA is on its way to accomplish even more than that.

Hello Dr Franko! Can you tell us what your role involves as Executive Director of Health Research Alliance (HRA)?

HRA’s mission is to increase the impact of HRA Members’ funding for biomedical research. I facilitate members’

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with members and other organisations with shared goals. I also lead HRA’s communications and its financial management.

for research and training in the field, address issues key to accelerating its development, and help the research community communicate with funders.

Can you also tell us about your additional role as a board member for the Center for Open Science?

As a member of the Center for Open Science Board of Directors, I promote the COS mission, help COS meet all legal obligations based on state law and the by-laws of the board, and maintain financial transparency and accountability for the organisation. We also review annual financial statements making sure that the financial management is sound and we approve the budget.

HRA is a strong believer in the value and robustness of COS’s open, public goods

How influential has the HRA been on investment in biomedical research?

HRA does not formally advocate but our members do. The Alzheimer’s Association, for instance, has had a major impact on investment in Alzheimer’s research. They have very generously shared their successful strategies with the rest of the HRA community and some members can already point to increases in funding in their own research space.

A rising tide lifts all boats so the fact that we speak with one voice about the importance of biomedical research and work with universities, the NIH, the FDA,

and other federal organisations helps to demonstrate the value of investment in biomedical research.

Can you tell us more about the HRA’s current plan of action to achieve its mission?

The HRA has a very comprehensive strategic plan. We host semi-annual Members’ Meetings which are the principal venue by which HRA members

interact, learn, network, and collaborate, and are especially valuable in enabling deep exploration of important issues in basic discovery and translational research.

HRA also has six working groups dealing with common problems in biomedical research like making science more ‘open and reproducible’, introducing new drugs and therapies, awarding grants, and all kinds of issues young researchers are

experiencing at the onset of their careers.

Another HRA’s initiative is The HRA Reporter, a real-time, searchable database of awards made by our members.

Currently, the database represents over $10 billion in funding and over 35,000 separate grants from 2006 to the present. Additionally, we use this database to publish reports, like the 2014 report “Bringing Non-Profit Funding

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of Biomedical Research Into the Light” which provided an enlightening analysis of grants for biomedical research from foundations and grant-making charities.

Finally, we publish e-newsletters and other updates to members and the external community about our initiatives,

The early involvement and leadership of BWF not only was pivotal in creating HRA, but the affiliation with the Fund provided HRA with legitimacy in its early days. The Burroughs Wellcome Fund Board and staff were also instrumental in encouraging private funders of

to enable members open and public access policies, and grantees’ access to an HRA-branded data repository. We have also developed the only database of non-profit funding of biomedical research, so the inclusion of organisation’s awards in the HRA Reporter database is valuable.

We need to make the public understand that investment in biomedical research is an investment in the future of our kids and grandkids highlights from the Member’s meetings, items of interest available on the website, information on upcoming meetings, webinars, and other activities and other information of interest to the membership and the broader biomedical community.

Which organisations have contributed to the and establishment of the HRA and support of its mission?

The engagement of the Burroughs Wellcome Fund, for example, was extremely important in the early days of the HRA, and BWF continues to be one of our very engaged members.

biomedical research and training to collaborate and to share information which resulted in the formation of HRA in 2005.

In addition to increased financial support in the form of ad hoc grants and other support mechanisms, staff members from organisations such as: the Doris Duke Foundation, the Damon Runyon Cancer Research Foundation, the American Cancer Society, the Donaghue Foundation, the Alzheimer’s Association, Foundation Fighting Blindness and many more, contribute countless hours of their time toward advancing the HRA mission.

This is on top of their very demanding day jobs!

How does the HRA compare to other organisations of nonprofit research funders? What makes the HRA different?

HRA is very action-oriented, so at HRA Member’s Meetings, and via the Interest Groups members work together to create valuable resources. We are wonderfully helpful peers – and there is easy access to other HRA members to compare policies and procedures and learn from each others’ experiences.

We also provide infrastructures such as an ORCiD consortium, a portal

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Organisation’s award data becomes part of an aggregate analysis of philanthropic biomedical research funding which is an important resource for HRA members for landscape analysis and benchmarking, but also for the broader scientific community. Membership also comes with free participation in two Members’ Meetings a year that cover both cuttingedge science and policy topics, and nuts & bolts advice.

Do you think biomedical research receives as much funding as it should and what are ways of improving this?

Of course, biomedical research could never receive as much funding as it should. Funded generously or even adequately, it could not only significantly improve human health and longevity around the globe but can also increase our quality of life. It also has the potential to be an economic driver for not only communities but for individuals as well.

We need to recognise the value of robust and rigorous science and put in place infrastructure to encourage and reward thorough science. We need to celebrate science and scientific researchers.

Philanthropy, researchers, academic institutions but also the government should play an important role in this celebration. Celebrations of scientific accomplishments and accomplished scientists should be as widely publicised as celebrations of teams winning the Super Bowl or the World Series. We also need to help scientists become better at communicating the value of science.

Everyone should be able to name an influential scientist (still living!) just like everyone can name an athlete or an

Thought Leader © Michelle Mishina

HRA doesn’t fund research itself, so we don’t have a funding strategy. But I would like to see PhD scientists trained and opportunities created so that they can use their training to contribute to the advancement of human health in ways other than as academic professors.

I also would like to see widespread open access to publications and other research outputs, as the ability to access and reuse research data, materials, and other research products have become even more vital to advancing science.

HRA's annual member's meeting

actor. If the general public recognises the value of biomedical research then we won’t have to fight as hard every year to increase budgets for NIH, NSF and other funding for biomedical research.

This is not something that the US should skimp on. For starters, we need to help the American public understand that investment in biomedical research is both an investment in the US economy and an investment in the future of our kids and grandkids.

From a personal perspective, are there any achievements you are particularly proud of?

When I was a member of HRA (long before I was the Executive Director), I co-founded HRA’s Early Career Scientist working group, which has evolved into the Research Workforce and Early Career Development Working Group.

There are so many thorny issues that affect the research workforce and HRA members are tackling many of these by implementing strategies and measuring outcomes. These are very hard issues (lack of diversity especially in senior positions, lack of opportunities in the academic setting, physician-scientist shortage, inappropriate reward structure especially

in academia, flat NIH funding and low success rates especially for early career investigators, training that doesn’t train for 21st century job landscape, etc.,) but HRA along with HRA members are working to have an impact where we can.

I am also very proud to be part of HRA’s very important Open Science efforts. I personally believe that funded research needs to be open so patients and families can make informed decisions, and boards/donors are able to evaluate the impact of funding. Even more critical is the requirement that resulting data be published in an easily accessible and machine-readable format to enable reuse and analysis by other researchers. Only then can the impact of the funding be multiplied – increasing the potential for significant and far-reaching advances and scientific innovation. HRA’s Open Science Task Force is making great strides in providing advocacy and resources to organisations that wish to and can move toward funding science that is more open.

What are your hopes for biomedical research improving human health in the future and how will HRA’s research funding strategy play into this?

One last area where I think HRA might have an impact is on Health Services Research. In an Annals of Internal Medicine article, the authors determined that “On the basis of contemporary data from 2009 to 2013, the median age of survival of patients with Cystic Fibrosis in Canada was ten years greater than in the United States (50.9 vs. 40.6 years, respectively). The adjusted risk for death was 34% lower in Canada than the US”. This is disturbing and something that is not addressed by traditional biomedical research, but is an area where we might be able to leverage contacts and resources. This is a new initiative so how much we can impact this area remains to be seen. HRA hopes to address the disparities in outcomes linked to address and not genetic code.

• To find out more information about HRA and their tremendous support for biomedical research, please visit their website at www.healthra.org.

Maryrose Franko, PhD Health Research Alliance P.O. Box 13605 65 T. W. Alexander Drive Research Triangle Park, North Carolina 27709 USA E: maryrose@healthra.org T: +1 240 393 2968 W: www.healthra.org

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COMMUNICATION

Social media and the rise of video Since the innovation of Tim Berners-Lee’s World Wide Web and the introduction of Google in 1998, the successive trend of online development has been in social media. Patrick Bawn takes a look at how important using video has now become across these platforms, especially within science.

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ith the world living in a time of online interactivity, and people becoming more and more immersed in using social media platforms, the landscape of human interaction has been forever changed.

The millennial age and the rapid emergence of technology has seen social media platforms becoming increasingly recognised as the go-to place for news and keeping up to date. On Facebook alone, there are 20,000 people online every second, with the average American spending around 40 minutes a day on the site. Recent statistics also show that Twitter has almost 320 million active users per month – representing a huge scope for potential interactions.

Not only that, but since YouTube’s inception in 2005, video has really taken off as a mainstream marketing channel, engaging audiences during the time that they spend online. You only need to check

your Facebook newsfeed to witness firsthand the number of videos posted online each day. In fact, on YouTube alone, there are over 300 hours’ worth of video uploaded by users every minute.

SENSATIONAL SCIENCE This extensive emergence represents an opportunity to bridge the ever-growing gap between researchers and the general public. Due to the accessibility and plethora of information now available online, science must engage fully with this medium in order to be represented in a way that is accurate – telling the story as it is meant to be told.

Sensationalised articles based on limited research are easy to come by with the growth of the internet, so it is important for science to move alongside this technological expansion. Becoming familiar with the methods in which videos can be used on social media could help to attract a new demographic of widespread,

It is time for the scientific community to embrace videos, making true, accurate research accessible to everyone

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tech-savvy consumers – ultimately aiding scientific research in the long run.

BRIDGING THE GAP Nowadays, watching videos through social media channels is the main method used by people to view content and access stories. It is time for the scientific community to embrace this method as well, making true, accurate research accessible to everyone – in a simple and engaging format.

For more information on turning your research into a simple yet effective animation, please visit our sister company SciAni at www.sciani.com.

Social Media for Scientists RSM was born out of multiple conversations with researchers who see a real benefit in connecting with a broad audience over an ongoing basis. Social Media can now be considered one of the most prominent and important engagement tools of the modern era. We help you get the ball rolling and can even provide long term Social Media Management support.

Start your Social Media journey now: www.researchsocialmedia.com

Partnership enquiries: simon@researchoutreach.org Careers and guest contributions: emma@researchoutreach.org www.researchoutreach.org