ethical aspects - German Stem Cell Network

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Applied Stem Cell Research in Germany

Annual magazine of the GSCN

regenerative medicine

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ethical a

translation

regulatory affairs

drug screening

2015/16

disease modelling

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cell therapies

drug screening

em cell

preclinical research

organoids


modelling stem cell banks creening genetic engineering drug screening stem cell bank

ethical aspects ISSN (Print) 2198-7831 ISSN (Online) 2198-784X

IMPRINT German Stem Cell Network e.V. Annual Magazine 2015/16 © 2016 GSCN Publisher German Stem Cell Network c/o Max Delbrück Center for Molecular Medicine (MDC) Robert-Rössle-Str. 10 13125 Berlin Tel.: +49 30 9406 24-87/-88 Fax: +49 30 9406 2486 E-Mail: [email protected] Web: www.gscn.org Editors Stefanie Mahler, Dr. Daniel Besser, Ulrike Papra (Central Office GSCN) Dr. Philipp Graf (BIOCOM AG) GSCN Pictures were taken by Yan de Andres and Stefanie Mahler Translation English Express, Berlin Design & Layout Unicom Werbeagentur GmbH, Stephen Ruebsam Print Buch- und Offset-Druckerei H. Heenemann Circulation: 1,100 Copyright The magazine is part of the public relations work of the GSCN and funded by the German Federal Ministry of Education and Research (BMBF). It is supplied for a protective fee of 2€ (inclusive postage) and must not be resold. Reprint only with permission by the editors. Disclaimer: As with all writings of GSCN people designations in favor of a better reading fluency are always gender neutral. ISSN (Print) 2198-7831 ISSN (Online) 2198-784X Applied Stem Cell Research in Germany

WELCOME NOTE THOMAS RACHEL

Welcome Note

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tem cells have become indispensable in biomedical research laboratories. Technical progress enabled by induced pluripotent stem cells (iPS cells) and genome editing methods have opened up novel approaches in the study of diseases – unthinkable just a few years ago. They also open up new possibilities in the treatment of diseases. Diagnostic procedures based on stem cells are also becoming more and more relevant in the pharmaceutical industry. German stem cell researchers are substantially involved in these developments. It is therefore important to combine expertise and to consolidate dialogue amongst all players. This is why the Federal Ministry of Education and Research (BMBF) initiated the development of the German Stem Cell Network (GSCN) three years ago – and has agreed to fund it until 2017. On this basis, the GSCN has established itself as an important forum. The network is a central platform on a national level as well as an institution that serves as

Annual GSCN Magazine 2015/16

a contact point for international partners and industry. And it actively supports social dialogue. This is reflected by the scientific and strategic working groups, as well as the successful GSCN Annual Conferences and numerous public information events, for example the organization and realization of the first European UniStem Day in Germany in March 2016.

I’m particularly pleased about the focus of this year’s issue of the GSCN Annual Magazine, as it covers clinical applications of stem cells. Germany will play an important role in the way that patients can benefit from stem cell research in the long term. We are supporting this development with a funding program for innovative stem cell technologies for individualized medicine, which was initiated in the second half of 2015. I’m delighted with the response from the GSCN members to this call, and I hope that the network maintains its active commitment to science, industry and society.

Thomas Rachel, MdB Parliamentary State Secretary of the Federal Ministry of Education and Research

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PREFACE ULRICH MARTIN

Preface

The high quality of the presentations and posters at the extremely successful Annual Conference in Frankfurt showed us once again that German stem cell research can hold its head up high. Despite having less funding available to us than leading countries such as the U.S. and Japan, we have once again seen outstanding publications by German stem cell researchers. In 2014, for instance, Nature published a paper by Zsuzsanna Izsvák and her working group on naive-like stem cells, which won the publication prize at the GSCN Annual Conference in September 2015. In addition, research centers have evolved that have a critical mass

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and are increasingly able to bring highly promising young researchers back to Germany, or to motivate them to stay. Sina Bartfeld is a good example of this. After a period spent as a researcher in Hans Clevers’ working group in Utrecht, she left to take up a position as group leader in Würzburg. An ongoing key area of focus for the GSCN Central Office and the strategic working group on Funding Programs and Policies is the production of a white paper on how the situation of German stem cell research compares to that of other countries. Although we were delighted by the BMBF’s recent call for proposals for research on and with pluripotent stem cells, I must stress that the public funding available for our pioneering research field is far below the level of leading countries such as the U.S. and Japan. With the white paper, we want to make the public and policymakers aware of this shortcoming and draw attention to the danger that Germany could soon be relegated to the fringes of a field that is extremely promising from both a medical and economic perspective.

In general, stem cell research has achieved impressive and in some cases dramatic advances in recent years. My personal highlight was and still is the incredible progress being made with genome engineering based on TALEN and CRISPR/Cas technologies. Following Shinya Yamanaka’s Nobel Prize for the discovery that mature cells can be reprogramed to become pluripotent, CRISPR/Cas technology, which is extremely relevant to stem cell research, is now also being hotly tipped for a Nobel Prize. Along with the development of iPS technology and scalable culture technologies, as well as the increasingly efficient differentiation of stem cells into a wide variety of cell types, this key technology has helped to get large pharmaceutical companies interested for the first time in using stem cells. We are also starting to see signs that clinical applications of adult stem cells and cell products, manufactured from pluripotent stem cell lines, could become clinical reality in the near future. For me, it will be extremely interesting to see whether high-cost autologous therapies that are highly complex in terms of production and characterization will actually win

Photo: GSCN

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ith the German Stem Cell Network (GSCN) now in its fourth year, we feel this is a good time to take a look back. If we weigh up our performance so far, we see no reason to doubt that our first years have been a resounding success. With financial support from the German Federal Ministry of Education and Research (BMBF), we have been able to build up an extremely vibrant network, which now enjoys a high degree of respect across all research locations in Germany. In addition to keeping our overall membership numbers stable, we have also attracted an increasing number of institutions actively involved in stem cell research to join the GSCN. In addition, we have welcomed a growing number of members from other European countries over recent years. The activities of the various GSCN working groups, the establishment of a network of German pluripotent stem cell core facilities, Pluricore, and the involvement of individual members in, for instance, the “meet the expert” tables during our annual conference show how willing our researchers are to be actively involved in shaping the GSCN’s development. Another achievement that is surely particularly worthy of note this year was the first-time participation of German stem cell researchers in the international UniStem Day that will be held in March 2016.


out in the end – or whether scientists will find ways of overcoming the allogeneic immune barrier even when transplanting universal stem cell lines.

Of course, as with almost any new technology, there are important hurdles to overcome and risks that must be taken seriously. For instance, after observing genomic abnormalities in one of the iPS stem cell lines that they had produced, the team working in Japan on the first phase I clinical trial into using iPS derivatives to treat macular degeneration put a temporary halt to the trial. Their actions show how seriously stem cell researchers and the relevant regulatory authorities take these types of risks. Irrespective of the type of cell, genetic abnormalities can never be ruled out in cases where cell expansion happens on a large scale. However, no other cell source has so far been investigated for possible genetic abnormalities as extensively as pluripotent stem cells have. I personally believe that the scientists involved and the physicians responsible possess the foresight not to endanger the future of stem cell-based therapies by behaving recklessly.

Photo: MHH / Henning Kempf

We chose translation and applied research as the focus of this year’s Annual Magazine. The following pages should give you a wide-ranging insight into topics related to the application of stem cells in research and therapy development. Individual chapters will address cell therapies, key regulatory issues, the understanding of disease models, application examples for testing active substances and stem cells in preclinical research, and the ethical aspects and problems of untested stem cell therapies. I would like draw your attention to the GSCN’s Annual Conference in September of this year. It will be held at Hannover Medical School in collaboration with the REBIRTH cluster of excellence. The event will focus on genome engineering as well as the many facets of clinical translation. I can promise a large number of leading international stem cell researchers and the chance to attend a wide range of outstanding presentations by junior researchers.


Cardiomyocytes derived from human pluripotent stem cells

In closing, I would like to express my sincere thanks to our many active members and to the Extended Board of the GSCN for their outstanding dedication. In particular, I am extremely grateful to my predecessor, Thomas Braun, for his exemplary commitment, and to the staff at the GSCN Central Office for their excellent work; without them the GSCN would not be such a success. The detailed activities of GSCN e.V. are included in the GSCN Annual Report, which you will find in this issue of the magazine. I hope that you share my enthusiasm for the great progress GSCN has made over the past four years, and that you find the latest issue of our Annual Magazine an interesting and enjoyable read. Best wishes,

Ulrich Martin President of the GSCN

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INDEX

Welcome Note Thomas Rachel (BMBF)�� 3 Preface Ulrich Martin, acting GSCN-President�� 4

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A network that provides impulses Interview with Ulrich Martin and Daniel Besser�� 8

GSCN-News from Central Office�� 12

Stem cells, mini organs and translation 3rd Annual Conference of the GSCN �� 16

12 16

Regenerative Medicine 22

Cell therapies on trial

22 Interview with Egbert Flory 28

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Stem cells need flexible rules

Stem cell-based disease models 34

Organoids: 3D miniatures are booming 6

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INDEX

38

Stem cell-based drug screening 38

On the hunt for new drugs

Preclinical stem cell research 43

Good models needed

48

43 Ethical debate 48

Controversial cuts

Essay: Untested stem cell therapies 52

Quacks and charlatans

52 54 GSCN Annual Report 2016�� 54

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GSCN | INTERVIEW MIT THOMAS BRAUN UND ANDREAS TRUMPP

Talking about the GSCN: Daniel Besser, managing director of the GSCN (left), and acting president Uli Martin.

Uli Martin is the new president of the German Stem Cell Network

“A network that provides impulses”

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INTERVIEW WITH ULI MARTIN AND DANIEL BESSER

The GSCN is now in its fourth year, and its fourth president is in office. The first three years were very much about founding and establishing the network, and overcoming the obstacles that came along with that. Now it is time for the network to direct its sights into the future and define its goals and strategies in greater detail, based on what has already been achieved. An interview with new GSCN president Uli Martin and GSCN managing director Daniel Besser on the network’s plans, visions, hopes and challenges.

Photo: GSCN

GSCN Annual Magazine: The German Stem Cell Network (GSCN) has now existed for almost four years. What, in your opinion, are the strengths of the network, and in what areas should it be doing more?

per will make it clear to policymakers how highly stem cell research is regarded in other countries and that it is in the government’s interest to strengthen Germany as a research location.

Uli Martin: That’s why I’m so pleased that the network exists. As an individual researcher, one often has the opportunity to exert influence on the regional or even the national level – either as an expert, via personal contacts, or as part of advisory committees. But when we get together as a network, we can have a much greater impact. In the coming year I will direct much of my efforts into making sure our voice is heard more clearly.

Uli Martin: We have already achieved a great deal. A large What is your current assessment of the situation for number of German stem cell researchers are members of the stem cell research? What topics in the field do you find GSCN, and we also have a significant number of members most interesting at present? from other European countries. Our conference has become an important event on the scientific calendar each year, and Uli Martin: I am fascinated by the incredible pace of devewe have some good ideas about how to communicate and lopments in genome editing – both with TALENs and with CRISPR/Cas9. That topic will keep us very busy for a while. network further, and how to put even more visions into Clinical translation is another very important aspect practice. However, what we don’t yet have sufficient for me. At the Hannover site, where I am based, political weight. That is evident from the rela„When we we are very close to the Medical School, and I oftively low support stem cell research is receiving get together ten see that both researchers and medical pracfrom public funding bodies in the international as a network, titioners have a long way to go in ensuring that comparison. I was delighted when the Federal we can have our findings are translated into practice more Ministry of Education and Research issued a a much greater effectively and that we collaborate better in this call for proposals last year. For me, that was the impact.“ regard. first sign of the turnaround we need. NonethelI think the time has now come to focus our attention ess, I would like to position the GSCN as a network more closely on planning future clinical applications. It that provides impulses to the political decision-making process relating to stem cell research and in getting better is important that we take this step now, as the clinical implefinancial support for stem cell research in the medium term. mentation of highly promising developments from basic research urgently requires comprehensive studies on how we Daniel Besser: I absolutely agree, and I am looking forward can overcome existing technical hurdles, on risk assessment, to working together on channeling our energies into policy and on how to implement other regulatory requirements. In work. In 2016, the GSCN will release a white paper on the many research fields – for example, the heart, the eye, diafunding situation for basic stem cell research in a range of betes – we are clearly on the cusp of being able to put our different countries including Germany. It is remarkable how research into practice. little government support – both financial and legislative – At this point, we should also make it clear that we can test stem cell research receives in this country compared to other many elements of stem cell-based therapeutic concepts on countries such as Japan, Sweden, the U.K. and the U.S. Since animal models but that, ultimately, we will not be able to we are not interested in reigniting the debate on the use of properly assess risks or see if stem cell therapies can work human embryonic stem cells, our focus lies also on the finan- successfully until they are applied to human patients. Clinical cial aspects. We would like the government to demonstrate trials are also important for us to be able to draw conclusions more commitment to our research and to provide financial on what we should be focusing our research in the future. support. Indeed, we are receiving the relevant signals from The progress we have made so far has of course stirred up various quarters, and I hope and expect that our white pa- hopes, but we mustn’t expect too much.


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Daniel Besser has been serving as GSCN Managing Director since 2014. He was a cofounder and coordinator of the GSCN in 2012 and coordinated the establishment of the structure of the network in 2013. Besser studied biology at the Freie Universität, Berlin and holds a doctor‘s degree in biology from the University of Basel. He received his postdoctoral training at Rockefeller University where he focused on signal transduction, cancer research and stem cell biology. From 2004 to 2011 Besser headed a research team on stem cells at the Max Delbrück Center for Molecular Medicine, Berlin. As part of this activity, he attended a host of events and conferences on stem cells as speaker and delegate and organized events targeting different aspects of stem cell research.

How can the GSCN help promote translation even more?

first time in 2015. Of course, we’ll be continuing those in 2016. A panel discussion on translation would certainly be interesting for many conference visitors.

The REBIRTH Cluster of Excellence

Photo: GSCN

Daniel Besser: I strongly believe the GSCN must do even more to facilitate communication and exchange between clinicians and basic researchers. We must take action here – translating basic research into practice is an Do you have any new ideas this year on how important topic that is gaining in significance all to make the network more attractive to „We place the time. For instance, at the next conference I stem cell researchers? particular can imagine offering more sessions on this toemphasis pic, or perhaps even a central panel discussion. Uli Martin: The GSCN conference was a great on translational What do you think about that, Uli? success, but I would still like the network to research.“ offer more to its junior scientists. One idea is Uli Martin: That’s very interesting, of course, for a winter or summer school on stem cells. It especially since the 2016 conference is taking place would be an intensive week-long event full of learnin Hannover and Hannover Medical School has always ing and shared experiences and an unforgettable opplaced particular emphasis on translational research. At portunity to make new contacts and to share news and the 2015 conference in Frankfurt I welcomed the fact that views with “big shots” and other colleagues in a wonderthe structure of the conference was changed so that the ful setting. Presidential Symposium became one central element. The presentations were excellent, and it was good that the Daniel Besser: Sometimes we do find that although the session had such a central place in the program. Perhaps GSCN offers many things – the Travel Awards, the workwe could do something similar in 2016 with the focus on ing groups, the sessions at the conferences, the annual translation. magazine, etc. – in some cases members are not that committed to participating. For example, we have set up Daniel Besser: The presidential session arose out of the a Humhub social intranet as a forum for exchange in and new GSCN Scientific Awards, which we presented for the among the various working groups, but that hasn’t really

From Regenerative Biology to Reconstructive Therapy REBIRTH has, under the nationwide Excellence Initiative, been funded as a cluster of excellence since 2006. The aim of the internationally renowned centre for regenerative medicine is to develop innovative therapies for the heart, liver, lungs and blood, and to translate these into clinical use. This involves collaboration – in Hannover and at participating partner institutions – between physicians, physicists, chemists, biologists, engineers, legal professionals and ethicists, the main research priorities being stem cell biology, the reprogramming of cells for cell therapy, disease models and tissue engineering. 10

Participating Partners: • Hannover Medical School • Leibniz University of Hannover • Hannover Laser Centre • University of Veterinary Medicine Hannover, Foundation • Helmholtz Centre for Infection Research Braunschweig • Max Planck Institute for Molecular Biomedicine, Münster • Institute of Farm Animal Genetics, Friedrich Loeffler Institute, Mariensee • Fraunhofer Institute of Toxicology and Experimental Medicine, Hannover

Exzellenzcluster

REBIRTH Cluster of Excellence Hannover Medical School Carl-Neuberg-Straße 1, 30625 Hannover www.rebirth-hannover.de



Ulrich Martin is full professor for Cardiorespiratory Tissue Engineering, head of the Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO) and director of research of the Clinic for Cardiothoracic-, Transplantation and Vascular Surgery (HTTG), MHH. His research basically focuses on induced pluripotent stem cells, the differentiation of pluripotent stem cells into cardiovascular and respiratory lineages, regenerative cell therapy, genetic engineering, and cardiovascular tissue engineering. Since September 2015, Martin has been the acting president of the German Stem Cell Network (GSCN). He is author of more than 100 publications in renowned international journals.

Daniel Besser: Each year, we hold two large events for the interested public. We produce videos on the various aspects and trends of stem cell research and post them online. We work closely with European organization EuroStemCell. And we organize numerous school events. Lots of things are going on behind the scenes. One major event will be UniStemDay on March 11, 2016. Uli Martin: Of course, many new ideas often „One major That’s a nationwide info and discussion event take time to get going and gain momentum. I event will be on stem cell research involving ten institutions can well imagine the Humhub social intranet UniStem Day on in seven locations. More than a thousand school becoming a lively communication platform in the coming year. Personally, I often just don’t 11 March 2016.” students will be attending. It’s a really big deal. have the time to get to grips with new online Uli Martin: I think it sounds fascinating – school communication options. But I still think it’s an students and young university students across Europe important idea. For our junior scientists in particular, Humhub is a terrific platform for the direct exchange of meeting for a day to find out more about stem cell research. It’s nice to see how we’re proactively and constructively experiences and reports. working all the fields. Especially when we don’t forget to Daniel Besser: Especially because we always post pro- occasionally combine social networking with actual sociamotional codes and announcements on Humhub first. lizing. Some of the absolute highlights for me last year were Which means that Humhub members have immediate the two GSCN parties. There was the WunderBar night at the ISSCR Annual Meeting in Stockholm, with a wonderaccess to all the most exciting information. ful view over the city, and the party at our conference in Frankfurt. Now I’m counting on having just as much fun in Hannover! What ideas does GSCN have for better informing the public about stem cell research? Interview: Stefanie Mahler

Photo: GSCN

gotten going as yet. What’s probably lacking is the group experience of a summer or winter school, which would probably generate a team spirit that could flow into continued professional dialog between the members of the GSCN working groups.

Max Delbrück Center for Molecular Medicine (MDC) Located in Berlin-Buch, the MDC carries out high-quality, interdisciplinary research on basic mechanisms and applications in major human health threats including cancer, cardiovascular and metabolic diseases, and disorders of the n ervous sytem.

These thematic research areas are supplemented by the Berlin Institute for Medical Systems Biology (BIMSB) at the MDC and the MDCCharité partnership in the Berlin Institute of Health (BIH). Annual GSCN Magazine 2015/16

Professor Mathias Treier, senior group leader at the MDC, states: “Opportunities for partnerships with clinical groups, a range of cutting-edge technology platforms, and superb animal facilities for diverse model organisms make the MDC an excellent site for stem cell research.” Recently, MDC and BIH have created a stem cell core facility to offer expertise to derive and manipulate iPSC lines for MDC groups or BIH projects. Alongside assisting groups, the facility is putting an emphasis on training, says Sebastian Diecke, head of the facility.

Max Delbrück Center for Molecular Medicine (MDC), Berlin-Buch Robert-Rössle-Str. 10, 13125 Berlin www.mdc-berlin.de

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FROM GSCN CENTRAL OFFICE

GSCN-News GSCN moratorium on engineering of the human germline In view of the extraordinarily rapid development of genetic engineering techniques, especially CRISPR/Cas, the Board of the GSCN resolved in March 2015 to support a moratorium on genetic intervention in the human germline, and published a statement to this effect. The call to postpone the use of gene editing in the human germline was issued in light of the uncertain ethical, social and legal implications of manifest and long-term modification of the human genetic base. The statement on the moratorium can be found on the GSCN website. The International Summit on Human Gene Editing held in Washington in December 2015 also issued a statement on engineering of the human germline. The statement is largely in favor of genetic engineering of embryos and germ cells – although only for research purposes and not in order to induce a pregnancy. New: GSCN Awards

In 2015 the GSCN established three new awards for German stem cell research: the GSCN Young Investigator Award, the GSCN Female Scientist Award, and the GSCN Publication of the Year Award. The three winners, Julia Ladewig, Magdalena Götz, and Jichang Wang, delivered presentations on their work at the first-ever presidential session, held during the 2015 Annual Conference. The GSCN awards will be presented again in the coming year. Proposals can be submitted to the Central Office now.

cal translation. This has motivated scientists to exchange the techniques and protocols they have developed. The “Stem Cell Cores,” facilities at the different institutes, will establish common standards for experiments and exchange knowledge and materials. This will include expertise regarding techniques to reprogram stem cells, stimulate their differentiation, engineer genomes, and characterize cells, and scientists will exchange useful vectors and cell lines. The initiative was launched by Micha Drukker (Helmholtz Center Munich), Harald Stachelscheid (BIH Stem Cell Core, BCRT) and Sebastian Diecke (BIH Stem Cell Core, MDC) and supported by the BIH and the GSCN. PluriCore chose the GSCN Humhub (see below) as communication platform. New GSCN films in German and English In 2015 the GSCN produced two more new films covering potential applications of cell therapies – the films are available in German and English on our website www.gscn.org. They can be used free of charge for information purposes.

GSCN award ceremony: from left Julia Ladewig, Magdalena Götz, Zsuzsanna Izsvák, Jichang Wang and GSCN president Thomas Braun

Network of German Stem Cell Core Units „PluriCore“

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On 30 - 31 July 2015, scientists from labs throughout Germany that provide services in the field of PSC research (core facilities) met at the Berlin Institute of Health (BIH) to establish a network, PluriCore, to promote scientific and technical exchange as well as collaboration. A high degree of standardization and harmonization is required when working with pluripotent stem cells (PSC), especially when the work is directed towards clini-

Everyone wants to communicate. Now, we can do it via the GSCN Humhub (gscn.humhub.com), a modern communication platform with social media functions that the Central Office has set up for the network and the working groups. Users can communicate with each other, exchange texts and drafts, upload photos, and discuss papers and reports. What’s more, promotional codes for conferences will appear here first. It is currently rather slow to get off the ground in some working groups – so it is worth remembering that all GSCN members are automatically part of the network. If you need help logging on, Stefanie Mahler (stefanie.mahler@ mdc-berlin.de) will be happy to assist.

Photos: GSCN

Humhub: Social Intranet for all of us!



ISSCR 2015 in Stockholm Networking for GSCN members and guests – the WunderBar Evening Scientists travel to conferences all over the world to present and listen to presentations. Personal contact with colleagues is equally important. So at the ISSCR 2015 Annual Meeting, the GSCN invented something new: the WunderBar Evening. Its purpose is to allow researchers to meet, celebrate, talk, eat and drink – in short, to network. And that worked wonderfully in Stockholm in 2015, when around 100 GSCN members and friends met above the rooftops of the Swedish capital and spent the midsummer evening enjoying the sociable atmosphere and stimulating chat. Our plan proved a complete success, and we hope a second GSCN WunderBar Evening will take place at ISSCR 2016 in San Francisco.

Photos: GSCN

Lively attendance at the GSCN Meet-up Hub The GSCN Meet-up Hub has been a popular tradition at the ISSCR since 2013. Many GSCN members and their colleagues from all over the world meet up and get talking, making use of the hour-long occasion to make or renew contacts and exchange news and views. The tradition continued in 2015. Even German journalists turned up, looking to speak to experts and make contacts with compatriots.

21 Travel Awards: “A great and unique opportunity” Twenty-one young members received Travel Awards from their GSCN working groups to enable them to attend the ISSCR 2015 Annual Meeting. Their feedback was enthusiastic: they commented positively on the special atmosphere, the presence of several thousand scientists from all over the world, the presentations by experts, the options for initiating joint projects, and the interesting discussions and ideas at the poster presentations. Their inspiring reports can be found on our website.

Global politics and science in dialogue: the GSCN at the World Health Summit in Berlin Since it was first held in 2009, the World Health Summit has become one of the most important forums in international health research. Each October, medics, scientists, social scientists, politicians, and representatives of the private sector and non-governmental organizations gather in Berlin. In 2015 the GSCN organized the session “Stem Cell Research – Paving the Path to Application” A panel of experts discussed the current status of clinical applications of cell therapies and likely developments. Their conclusion was that in many areas including diabetes, neurological diseases and eye and heart disorders, research results are ready for testing in clinical trials. For further details see Daniel Besser’s essay “Stem cells in disease modeling and regenerative medicine – perspectives, challenges and realities” in the WHS Yearbook 2015.

From left to right: Ulrich Martin, Zameel Cedar, Mohan Vermuri, Egbert Flory, Daniel Besser, Fathia Sadallah, Arnd Hoeveler, Daniel Pipeleers, Casja Lindberg, Tim Allsopp, Joachim Müller-Jung

GSCN session at the DGTI in Basel At the 48th annual conference of the German Society for Transfusion Medicine and Immunohematology (DGTI, 15 – 18 Sept. 2015) in Basel, the GSCN organized a session on the principles of stem cell research, as it did in 2014. The workshop was opened by Benedikt Berninger (Mainz) with his talk “Remodeling and reprogramming the adult brain.” Berninger showed that the formation of presynaptic inputs

Fraunhofer Institute for Cell Therapy and Immunology Fraunhofer IZI

The Fraunhofer Institute for Cell Therapy and Immunology IZI investigates and develops solutions to specific problems at the interfaces of medicine, life sciences and engineering. One of the institute’s main tasks is to conduct contract research for companies, hospitals, diagnostic laboratories and research institutes operating in the field of biotechnology, pharmaceuticals and medical engineering.

The Fraunhofer IZI develops, optimizes and validates methods, materials and products for the business units Drugs, Cell Therapy, Diagnostics Annual GSCN Magazine 2015/16

and Biobanks. Its areas of competence lie in cell biology, immunology, drug biochemistry, bioanalytics and bioproduction as well as process development and automation. In these areas, research specifically focusses on the indications oncology, ischaemia, autoimmune and inflammatory diseases as well as infectious diseases and regenerative medicine. The institute works in close cooperation with hospital institutions and performs quality tests besides carrying out the GMP-compliant manufacture of clinical test samples. Furthermore, it helps partners obtain manufacturing licenses and permits.

Fraunhofer Institute for Cell Therapy and Immunology IZI Perlickstr. 3, 04103 Leipzig www.izi.fraunhofer.de

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World Conference on Regenerative Medicine (WCRM) and GSCN satellite event in Leipzig

From 21 – 23 Oct. 2015, more than 800 scientists met in Leipzig for the World Conference on Regenerative Medicine, an event that has been held in the Saxon city every two years since 2007. For two satellite events, the GSCN working group “Technologies in stem cell research,” headed by Andreas Bosio (Bergisch-Gladbach) and Frank Emmrich (Leipzig), put together an excellent program with six outstanding speakers:

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• Walter Brehm, Leipzig University, Germany: “Stem cell techniques in veterinary medicine from clinical application to basic science” • Jonathan Bramson, McMaster University, Canada: “Engineering white blood cells to kill cancer” • Nicola Elvassore, University of Padova, Italy: “Endogenous signaling pathways control reprogramming and differentiation in microfluidic environment” • Agnete Kirkeby, Lund University, Sweden: “Bringing hESCs to the clinic for treatment of Parkinson’s disease” • Harold Cremer, IBDM, Marseille, France: “Molecular control of neural stem cell determination in postnatal brain”

From left to right: Walter Brehm, Jonathan Bramson, Frank Emmrich, Harold Cremer, Agnete Kirkeby, Nicola Elvassore, Johnny Kim, Daniel Besser, Andreas Bosio

• Johnny Kim, Max Planck Institute, Bad Nauheim, Germany: “Functional systems analysis of prospectively purified adult muscle stem cells” The GSCN’s European activities: Networking with EuroStemCell

The GSCN is a German partner of the international information portal EuroStemCell, an association of European stem cell research centers. The GSCN has translated many EuroStemCell materials into German, including factsheets containing up-to-date information on stem cell research relating to diabetes, cancer, leukemia, eye diseases and ethical issues, as well as stem cell games for use at public outreach events. The materials, which are suitable both for events and for answering patients’ queries, can be found on the website www.eurostemcell.org and will shortly also be available for download as a PDF from the GSCN website www.gscn.org Stem cell research for outstanding students Bright-minded students visit research institutions: Students of STEM (MINT in German) subjects are interested, curious and very well educated. In February 2015, the GSCN took part in the capital forum MINT 400 in Berlin, meeting keen and open-minded students from all over Germany who enthusiastically tried out new things in the laboratory and engaged in scientific and ethical discussions about stem cell research. UniStem Day – a European stem cell day

In 2016, research institutes across Germany will for the first time take part in UniStem Day – a European day of stem cell research. On 11 March, school students in eight cities will be able to participate in education days organized by the GSCN – with guided visits, presentations, meet-the-scientist tables, games, films, and video transmission between European cities. More than 1,000 senior high-school students specializing in biology at schools in Berlin, Bochum, Bonn, Dresden, Hannover, Heidelberg, Jena and Münster are expected to participate. Last year, 25,000 young people across Europe took part in UniStem Day. For more information visit www.unistemday.de

Photos: Fraunhofer IZI; GSCN

in newly generated neurons in the adult hippocampus is strongly dependent on experience during a critical period two to six weeks after generation of the neuron. He also discussed data that prove that some neurons generated from glia cells by reprograming through the use of transcription factors integrate into the pre-existing neural network. In a presentation entitled “Functional profiling of AML stem cells,” Frank Buchholz (Dresden) described data that explain the molecular mechanisms of cancer stem cells in acute myeloid leukemia (AML). Using a comprehensive test system involving interfering RNAs (iRNA), it was possible to identify six genes that are important for the growth and survival of cancer stem cells. Blocking a gene product known as ROCK1 kinase resulted in effective and specific elimination of the leukemia cells, also in a humanized mouse model. By means of single-cell analysis and elegant use of color-marked fusion proteins in mouse lines, Philipp Hoppe (Basel) showed in his talk “Transcriptional control of hematopoietic lineage choice – old paradigms and new insights from single-cell analysis” that the decision on differentiation into either megakaryocytes/erythrocytes or granulocytes/monocytes does not depend on the random fluctuation of two transcription factors, PU.1 and Gata1. These experiments also identified new myeloid hematopoietic sub-cell types. In her presentation “Analyzing human hematopoietic stem cell biology in vivo: Cell cycle progression as mediator of cell fate decisions”, Nicole Mende (Dresden) described data on the regulation of the G1 phase of the cell cycle of hematopoietic stem cells (HSC) by the factors cyclin D1 and the associated kinase CDK4 (cyclin-dependent kinase 4). She showed that shortening the G1 phase of the cell cycle has a beneficial impact on the functioning and maintenance of human HSCs.


GSCN | 2. JAHRESKONFERENZ


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3rd Annual Conference of the German Stem Cell Network 2015 from 9 - 11 September 2015

Stem cells, mini organs and translation

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or three days, more than 400 German and international researchers discussed all aspects of stem cells in poster sessions, after presentations or while networking during the breaks and evening events. The GSCN conference took place in a very attractive environment: the newly built Otto-Stern- Center has large auditoriums, a foyer for exhibitors and catering filled with natural daylight, and a sunny terrace.

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The conference got off to a vigorous start on Wednesday with opening addresses from Enrico Schleiff, Vice Chancellor of Goethe University Frankfurt, and GSCN President Thomas Braun as the patron. Paul Paul Riley (University of Oxford, U.K.) opened Riley the scientific program with a keynote lecture in which he presented new findings from his lab on epicardium activation using Thymosin ß4. Riley’s team has found that Thymosin ß4 interacts with a chromatin remodeling complex to express the Wilms’ tumor gene (Wt1), which also plays an important role in epithelial-mesenchymal transition of the epicardium. Moreover, a long

antisense oriented non-coding RNA produced by the Wt1 locus appears to play an important role in maintaining Wt1 expression. These processes are central to the epigenetic reprogramming of epicardium-derived cells and could lead to new approaches to treating heart disHans Willem ease. Hans-Willem Snoeck, from Snoek Columbia University in New York, reported in the second keynote presentation about his recent progress in modeling the developing human lung in a Petri dish using iPS cells. He presented highly convincing data on the specific differentiation of human pluripotent stem cells into foregut endoderm cells and especially into distal lung cells. Snoek showed that the sequential application of defined cytokines and low molecular weight compounds that activate or inhibit signaling pathways results in robust differentiation protocols. His work is a good example of the translation of knowledge from the basic principles of developmental biology in mice to the selective differentiation of human pluripotent stem cells. It shows that complex multi-stage organ development can be imitated in a Petri dish. The scientific sessions

One characteristic feature of the GCSN conferences is their function as a platform for young up-and-coming scientists, in particular, to present their latest findings. The best papers were selected for ten parallel sessions. The conference program therefore reflects current issues in stem-cell research, including nuclease-based techniques such as TALENs and CRISPR/Cas, which are precise new genome

Photos: GSCN

In September it was that time again: 400 scientists from all over Germany met for an intensive three-day meeting in Frankfurt am Main. 2015 was the third edition of the GSCN conference, which already has a strong tradition. But tradition is no barrier to progress in this young field of research and young network. A new development in the conference format, the central presidential symposium with presentations by the GSCN prize winners, was very well received by participants. Another positive development is the growing number of exhibiting companies – 25 firms showcased themselves with booths and lectures. The network is growing not only in numbers but also in significance.


Photos: GSCN

editing tools that can effect highly accurate changes in the One novelty and highgenome. The development of stem cells as disease models light of the GSCN conferin cell culture – including organoids (three-dimensional ence was the presidential mini-organs in a culture) – also played an important role. symposium, at which the Reprogramming and transdifferentiation of body cells, GSCN awards, inaugurated particularly neural cells, were also themes of the third an- just last year, were presented and nual conference. The whole conference program included the winners gave lectures. To kick off 42 presentations selected from 180 submitted abstracts, the symposium, GSCN Founding Presiincluding seven papers in the strategic working group ses- dent Oliver Brüstle gave a lecture entisions and nine presentations by partner companies. “The tled “Evolving stem cell-based strategies program is incredibly exciting and packed,” GSCN President for targeting neurological disorders.” He introduced new strategies for Braun enthused. “The quality and scientific level are using patient-specific stem cells very high. I get the impression that year on year we “An in researching and treating neugo deeper into the subject.” exciting rological diseases. His team used and packed neurogenic and gliogenic stem cells, These were some of the key highlights from the program” which are derived from induced plupresentations: In the “Stem cells in disease modripotent stem cells (iPS cells) or obtained eling and drug development” session, Sina Bartfeld (Hubrecht Institute, Utrecht, now at the Research Center by direct conversion from blood cells. Such stable infor Infectious Diseases [ZINF] in Würzburg) gave a pa- termediates allow highly standardized modeling of earlier per on organoids from adult stomach stem cells and their pathogenic processes, such as protein aggregation in polyuse in infection biology. Stem cells can be obtained from glutamine diseases and the preparation of patient-specific patient tissue samples, and can grow in vitro into small donor cells, including cell-based enzyme substitution in three-dimensional structures, or organoids, which can lysosomal storage diseases. Julia Ladewig of the Institute be (apparently unlimitedly) expanded in culture. Human of Reconstructive Neurobiology in Bonn, and winner of stomach organoids infected with the stomach pathogen the GSCN Young Investigator Award 2015, presented her Helicobacter pylori showed a strong inflammatory re- latest data on the human in vitro development of neural sponse. The strength of this response depends on the cel- stem cells in comparison to neural stem cells in vivo. The lular composition of the organoids. Organoids from tumor study describes a matrix for examining the relationships tissue can also be used in drug testing. In the session on between cell types derived from pluripotent stem cells and hematopoietic stem cells, Nicole Mende of TU Dresden comparable native cell types. Ladewig also presented reshowed that the fitness of human blood stem cells (HSCs) cent data on the production and standardization of self-ordepends on the length of the G1 phase of the cell cycle. She ganizing cortical organoids. This latest technology is pardemonstrated that specifically shortening the early G1 ticularly relevant when the complex cellular interactions phase transit in human HSCs made it possible for the blood that occur in human tissue are examined in the cell culture. “I am delighted to accept the GSCN Award, because it is stem cells to produce a greater number of mature blood important for us junior research group leaders cells over a prolonged period, after transplanto receive public recognition for our scitation into a suitable mouse model. It is entific achievements early on in our conceivable that in future, stem cell careers. For me, the GSCN Young function can be improved in the Investigator Award is a great human body by accelerating the signal that encourages me to cell cycle transit. Lina Jankausincrease my scientific efforts kaite (UGMLC, Gießen) reand shows me that I am on ported in the session “Stem the right track,” Ladewig encell in regenerative therathused. pies: Mesenchymal stem/ Magdalena Götz was also stroma cells” on the use of visibly delighted with her MSCs as a putative therapy GSCN Female Scientist for influenza-virus induced Award: “I feel very honored pneumonia. In a murine and am truly thrilled with model of influenca virus inthe GSCN Award.” In the lecfection intratracheal therature she gave after accepting py with bone marrow-derived the award, Götz presented new, stromal cells (BMSC) was shown groundbreaking data, showing that to reduce virus-induced injury of the efficiency of directly reprogramalveolar epithelial cells (AECs) and to ming fibroblasts and glial cells in vitro into increase virus clearance. Mechanistically, neurons by neurogenic transcription factors BMSCs reduced AEC apoptosis by secretion of stanniocalcin-1 in vitro corresponding to improved regen- depends largely on a successful transition from predominantly glycolytic to oxidative metabolism. Glial cells and fieration of damaged AECs in vivo.


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3. GSCN-JAHRESKONFERENZ

broblasts which fail to make this transition during a critical phase in the reprogramming process undergo cell death mediated by reactive oxygen species (ROS), which is referred to as ferroptosis. In contrast, reducing the formation of ROS and overexpression of Bcl2 promotes reprogramming. The enormous significance of these findings is emphasized by the fact that, as Götz showed, overexpression of Bcl2 also significantly promotes neuronal reprogramming of glial cells in the injured neocortex.

Daniel Besser, Managing Director of the GSCN, was very pleased with the success of the redesigned conference program. “The presentations were exciting and it is great that the presidential symposium is now a key element of the program for all participants,” he said. The majority of the conference participants agreed, and in an online survey gave very positive assessments of the program and the structure and quality of the presentations. The two poster sessions generated a great deal of interest and were particularly well attended. Intense discussions took place around many of the posters. The poster prizes, provided by GSCN member company Peprotech, were awarded to the following: • Chiara Cencioni (Goethe University Frankfurt): “Nitric oxide synthesis and Zeb1 transcription factor inactivation characterize very early mesendoderm precursors in mouse embryonic stem cells” • Nico Lachmann (Hannover Medical School): “Pulmonary macrophage transplantation employing HSC- or iPSC-derived cells as an innovative gene therapy approach in pulmonary alveolar proteinosis”

Photos : GSCN

The GSCN Publication of the Year 2015 Award went to Jijang Wang, a doctoral candidate in the Mobile DNA research group and his group leader Zsuzsanna Izsvák of the Max Delbrück Center for Molecular Medicine (MDC) in Berlin-Buch, with their publication “Primate-specific endogenous retrovirus driven transcription defines naïve-like stem cells” (Wang, J. et al, 2014, Nature, doi:10.1038/nature13804). They were able to demonstrate new ways of identifying naïve, and thus original human embryonic, stem cells. “I am deeply honored and touched to receive this award. I am very grateful that the German stem cell community has selected our publication as the best of the year,” Wang said on receiving the award. Group leader Izsvák was equally delighted. “My lab is working with repetitive, genetically identifiable but replaceable elements. This means that we are naturally asking different questions from the ones a ‘typical’ stem researcher poses. As newcomers, we are particularly proud that we could contribute something fundamentally new to the field of stem cell research. It was a wonderful journey with great colleagues. In a nutshell, we have determined how remnants of an ancient retrovirus family, which on first sight look like typical junk DNA, have taken on a new cellular function to regulate real pluripotency in human cells.”

Leibniz Institute on Aging - Fritz Lipmann Institute (FLI) Research for better aging

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on society can be minimized and the society’s future development will be enriched by the wealth of knowledge and experience older people possess. As one of 89 institutes of the Leibniz Association, the FLI is publically funded by the German Federal Ministry of Education and Research (BMBF) and the State of Thuringia. Leibniz Institute on Aging – Fritz Lipmann Institute (FLI) Beutenbergstraße 11, 07745 Jena www.leibniz-fli.de

Photos: GSCN

The Leibniz Institute on Aging – Fritz Lipmann Institute (FLI) has dedicated to biomedical aging research since 2004. More than 330 members from over 30 nations explore the molecular mechanisms underlying aging processes and ageassociated diseases. It is one of only two national research institutes on basic mechanisms of aging in Germany. The main aim of research at FLI is to delineate how aging leads to the development of tissue dysfunction and diseases in the elderly. If the understanding of the aging process contributes to the extension of healthy lifespan, the strains


Photos : GSCN

• Krishna Moorthy Sreenivasan (Max Planck Institute, Bad Nauheim): “A high-throughput RNAi screen identifies chromatin modifiers regulation muscle stem cell self-renewal and differentiation” • Ting Zhang (Max Planck Institute, Bad Nauheim): “Epigenetic control of muscle progenitor and adult stem cell homeostasis”

The scientific program of the conference concluded with a joint session with the LOEWE Center for Cell and Gene Therapy Frankfurt (CGT) and two more keynote lectures. Andras Nagy of the Lunenfeld-Tanenbaum Research Institute in Canada presented his latest results in the presentation on “Pluripotency in the artificial cell space.” He presented an inducible reprogramming system that has been successfully used to define intermediate molecular genetic stages during the somatic cell reprogramming process. This made it possible to characterize a new (F-iPS) subtype of iPS cells that maintaining pluripotency remains highly active. It may also are fundamentally different from the fully reprogrammed iPS be possible to evoke this situation in inactive metastatic cells. cell in terms of genetics, epigenetics and phenotype. Unlike Trumpp pointed out that the MYC reactivation in such cells (for example, in cases of inflammation) could lead to the the fully reprogrammed iPS cells, F-iPS cells can be proonset of metastatic growth, often years after apparduced on an industrial scale because of their great ently successful chemotherapy. potential for self-renewal. They can then be dif„It was a ferentiated according to the respective end cells required for use in regenerative medicine. Since wonderful journey The CGT organized excellent presentations in the final session: Lorenz Studer was unfortunately unwell, Anwith great dreas Trumpp of the German Cancer Research colleagues“ • Gergana Dobreva, Max Planck Institute for Center in Heidelberg stepped in at short notice as Heart and Lung Research, Bad Nauheim: “Reguthe fourth keynote speaker. He opened his presenlation of second heart field progenitor cells: discotation on “Stem cell dormancy and MYC” with an oververing key molecular players” view of the molecular and cellular basis of self-renewal and differentiation in hematopoietic stem cells (HSCs) and their • Johnny Kim, Max Planck Institute for Heart and Lung Research, Bad Nauheim: “Functional systems analysis of interaction with stem cell niches. He then presented the new the adult muscle stem cell identifies crucial regulators data supporting the concept of reversible dormancy (inacof muscle regeneration” tive sleep-like state) in HSCs during homeostasis and under stress, based on whole genome transcriptome analysis on • Michael Rieger, Goethe University and LOEWE CGT the single cell level. These investigations and detailed funcFrankfurt: “Hematopoietic stem cell fate decision contional analysis revealed that the oncogene MYC plays a key trol at single cell resolution” role in controlling the initiation and cessation of dormancy • Zoltán Ivics, Paul Ehrlich Institute, Langen: “The Sleein HSCs and embryonic stem cells. Finally, he presented exping Beauty transposon system for genetic engineering citing data indicating that deactivating the MYC oncogene in in stem cells” pluripotent epiblast cells in the embryo before implantation in the uterus initiates what is known as the diapause state. Spontaneous meet-the-expert tables Diapause is a temporary resting state that is the embryo’s response to signals from the immediate environment (e.g. Always on the lookout for improvements and innovations, during breastfeeding). This restricts all metabolic activity at this year’s conference some professors on the GSCN Exand protein production in the embryo, while the network for tended Board spontaneously decided to offer a number of meet-the-expert tables, where participants could talk directly to principal investigators. A lively lunchtime discussion culture developed, and some conference participants used the opportunity to discuss their research and scientific issues directly and informally with Andreas Trumpp, Claudia Waskow, Hartmut Geiger and Hans Schöler. “This was a very interesting and lively encounter,” Claudia Waskow said. Meet-the-expert tables with Sebastian Diecke and Ralf Kühn had been planned in advance by the technologies working group. They joined the wider debate and discussed burning questions relating to the genome editing techniques CRISPR/Cas9 and TALENs. “That was an exciting workshop, and I was pleased to see how many questions were thrown at us,” said Diecke (Max Delbrück Center for Molecular Medicine). Poster awardees


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GSCN | 3. JAHRESKONFERENZ

New stem cell genome editing techniques, clinical trials, translation, regulatory issues and career development in science and science-related professions were other topics at the strategic working group sessions. The GSCN conference in Frankfurt, like its predecessors, was evaluated very positively in the online surveys of participants and exhibiting companies. It was suggested, however, that the GSCN should do even more to increase collaboration between basic researchers and the clinicians who ultimately use the applications on their patients.

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“Good networks need good parties – and we know how to celebrate,” said new GSCN President Ulrich Martin (Hannover Medical School). On the networking evening, the scientific community rode in open-top buses through downtown Frankfurt to the Westend campus. A stroll past the colorfully lit lake and old university buildings took

the scientists into the grand wood-paneled banqueting hall where they enjoyed an evening of food, chats and dancing in a relaxed atmosphere. The evening celebrations and gatherings at the conference spoke for themselves. Wherever they were, at the summery barbecue or the GSCN Networking Evening, dressed up in the photo booth, at the sumptuous buffet, or bopping away on the dance floor, the stem cell researchers showed that they know how to party, thereby laying excellent foundations for personal exchange and collaboration. Podium discussion for the Frankfurt public The conference concluded with a public event entitled “Modern cell therapies – stem cells in the fight against heart attacks and blood cancer.” New German Stem Cell Network films were shown on the Westend campus, and the interested public had the opportunity to take part in a discussion with Andreas Zeiher and Hubert Serve (both of Frankfurt University Hospital and LOEWE CGT), and Thomas Braun. To a full house, Stefanie Seltmann (German Cancer Research Center, Heidelberg) moderated a lively podium discussion about the heart’s capacity for regeneration and the potential clinical use of cell therapies. The GSCN films about the potential application of cell therapies present Andreas Zeiher’s and Hubert Serve’s cardiology and leukemia research, and are available in German and English on the GSCN website.

Photos : GSCN

The drop-in booth was another format in the conference program that generated absorbing direct Q&A sessions. Egbert Flory (Paul Ehrlich Institute) and Natalie Mount (Cell Therapy Catapult, U.K.) briefed participants on regulation, translation and clinical trials. The theme of translation pervaded many discussions and presentations. Last but not least, the workshop of the strategic working group on public outreach, with Ira Herrmann and Tobias Cantz, confronted the conference participants with citations from websites offering unverified stem cell therapies. The participants were asked to deal with the citations and to evaluate the content. This was not an easy task, but the workshop reached its goal: on the terrace there were lively discussions about clinical trials, translation, dealing with patients’ expectations, and the gap between hopes and promises.


KAPITEL 1 | LEBENDER KOLUMNENTITEL

4th Annual GSCN Conference 12 – 14 September 2016 Hannover Medical School (MHH) www.gscn.org

International keynote speakers Peter Coffey (London) · Alexander Meissner (Cambridge) · Sean Morrison (Dallas) · Hiroshi Nagashima (Tokyo) · Peter Zandstra (Toronto) Presidential Symposium Thomas Eschenhagen (Hamburg), GSCN Awardees: Young Investigator, Female Scientist, Publication of the Year

Abstract submission deadline for oral presentations: 31 May 2016 Oral presentations chosen from the best abstracts Scientific sessions

Photos: Erlebnis-Zoo Hannover · Nik Barlo jr. · HMTG/Kirchner · MHH · GSCN/Chen

• • • • •

Pluripotency and reprograming Somatic stem cells and development Tissue engineering and organoids Hematopoietic stem cells Stem cells in development

• • • • • •

Stem cells in diseases: cancer stem cells Stem cells in regenerative therapies Stem cells in disease modeling and drug development Computational stem cell biology and systems biology Stems cells and ageing, genome stability and epigenetics Genome engineering and gene therapy

Program committee Daniel Besser (Berlin) · Thomas Braun (Bad Nauheim) · Tobias Cantz (Hannover) · Tilman Fabian (Hannover) · Ulrich Martin (Hannover) · Karl Lenhard Rudolph (Jena) · Claudia Waskow (Dresden)

Supported by


Collaboration with

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translation

regulatory affairs

drug screening


disease modelling

organo

Neurons directly derived from transformed skin cells 22

ethical aspects

Photo: Uni Bonn / Julia Ladewig

cell therapies

drug screening

m ell


organoids


modelling stem cell banks reening genetic engineering drug screening stem cell banks


s

REGENERATIVE MEDICINE

Regenerative medicine

Cell therapies

on trial

Healing with cells – that is the principle behind regenerative medicine. Stem cells have long been a routine therapeutic agent in some fields, such as leukemia medicine. However, the high hopes harbored in Germany and elsewhere that adult stem cells might prove useful in treating other diseases have rarely been fulfilled. Now, scientists are steadily improving their understanding of how adult stem cells work, with the result that promising applications are currently being tested for safety and efficacy in clinical trials. Increased knowledge and expertise in dealing with pluripotent cells have ushered in a new era in cell replacement therapy. Worldwide, patients with eye diseases, diabetes and myocardial infarction are now being treated with replacement tissue derived from pluripotent stem cells.

W

hether for cell and tissue replacement or to support regeneration within the body, stem cells are a promising source of new treatments. But translating laboratory findings into clinical practice is difficult, because cell-based treatments are highly complex and are usually tailored to the individual patient. In addition, the treatments need to meet strict regulatory requirements and production standards. As with other drugs, the safety and efficacy of stem cell-based treatments must also be tested in clinical trials. The clinical trials register clinicaltrials.gov contains details of several thousand trials worldwide that involve stem cells. Almost 200 of these are in Europe. The Paul Ehrlich Institute, which is responsible for approving clinical trials in Germany, states that 35 clinical stem cell studies are underway (see interview with Egbert Flory). In Germany, clinical research into cell-based therapies is being pursued at the translational centers for regenerative medicine, which were set up more than ten years ago by federal and state bodies and the German Research Foundation (DFG). For more information on this, see the 2013/14 GSCN Annual Magazine. The centers include the REBIRTH Cluster of Excellence in Hannover, the Berlin-Brandenburg Center for Regenerative Therapies (BCRT), the TRM in Leipzig, the RTC in Rostock, and the DFG’s Center for Regenerative Therapies in Dresden (CRTD). With the establishment of the LOEWE Center for Cell and Gene Therapy (CGT) as part of the state of Hesse’s excellence initiative, the Frankfurt am Main area has also become a stronghold of cell therapy research. Which cell type for which organ?

Regenerative medicine utilizes two particular properties of stem cells that promote healing. Firstly, both pluripotent and multipotent stem cells can be used to produce replacement tissue. In a range of disorders that include some eye


diseases (e.g. Stargardt’s disease), Parkinson’s, myocardial infarction and diabetes, specific celltypes are destroyed. These disorders are particularly suited to cell replacement therapy: the hope is that the engineered tissue can replace the lost organ functions and support regeneration. Secondly, stem cells are a sort of “living pharmacy” which emits a cocktail of growth factors and messenger substances that promote regeneration or have a beneficial effect on the immune system. But which cell type is best for which disease? Traditional cell therapies are based on adult stem cells, but a new generation of cell replacement therapies is currently emerging. Preparations based on human embryonic stem cells (ES cells) and induced pluripotent stem cells (iPS cells) are being tested worldwide. Blood stem cell therapy for leukemia

The old hand among stem cell therapies has been around for more than 50 years: blood stem cell transplantation is a firmly established practice in leukemia medicine. The technique uses blood stem cells obtained either from bone marrow donation or by taking peripheral blood stem cells from the patient’s blood or that of suitable donors. The more modern blood stem cell transplantation method involves mobilizing the hematopoietic stem cells in the bone marrow with drugs and drawing them into the blood. They can then be extracted and filtered out by means of a type of blood washing in a blood donation process that lasts about five hours. If the cells are introduced into the patient via blood transfusion after chemotherapy, they autonomously settle in the bone marrow niches, then reproduce and re-start the process of hematopoiesis. In time, therefore, a new immune system develops.

“Obtaining stem cells from peripheral blood has greatly simplified the process of donating bloods cells and significantly increased willingness to donate,” says Torsten Tonn, Professor of Transfusion Medicine at the Carl Gustav Carus Faculty of Medicine at TU Dresden. In recent years, Tonn has also witnessed a resurgence in bone marrow donation. “In many cases it results in fewer side effects and rejections,” says Tonn, who is also Medical Director of the

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German Red Cross Blood Donor Service North-East. According to the figures of the German Registry for Stem Cell Transplantation (DRST) there were 3,220 allogeneic and 3,241 autologous blood stem cell transplants in Germany in 2014 (an allogeneic transplant uses stem cells from a donor; an autologous one uses cells taken previously from the patient). Although stem cell therapy is now routine in the treatment of leukemia, it still has risks and side efMacrophages for fects, including rejection therapeutical use and infection. Doctors are derived from stem cells therefore attempting to Human pluripotent refine the concept further stem cell derived and to control the effects of cardiomyocytes the cell therapy. “Blood stem cell transplantation is increasingly being regarded as immunoconcepts based on MSCs are currently therapy, and one important trend being tested in numerous clinical trials. here is the move to modify the transplant in specific ways using biotechnology,” Experimental cell therapies for infarcted hearts says Tonn. He goes on to explain that a tailored transplant involves – among other things – filtering out unwanted or Stem cells in the bone marrow are also being used in the disruptive immune cells and increasing the quantity of oth- regeneration of infarcted hearts. German researchers are er cells to support the recipient’s immune defenses. among the pioneers of this experimental stem cell therapy. Combined stem cell therapy

In another trend, blood stem cell therapy is being combined with mesenchymal stem/stromal cells (MSCs). The special feature of these adult cell types, which can likewise be obtained from the bone marrow and from other tissue, is that they have immunomodulatory properties and can therefore suppress an overactive immune response in the body. Doctors hope to use the immunosuppressive function of MSCs to tackle a much-feared complication, that of graftversus-host disease (GvHD), in which the immune cells of the donor transplant attack the recipient’s cells, triggering potentially severe immune responses. Promising therapy

With hindsight, the first steps back in 2001 were not something to be so proud of. At that time, Bodo-Eckehard Strauer, a cardiologist in Düsseldorf, became the first person in the world to use patients’ own bone marrow cells in the treatment of heart attacks. The experiment was a bold one: the intervention had only been described in mice a few months previously. Other attempts followed, with promising results, but by then experts in the field were becoming highly critical of this overhasty translation into practice. It is now clear that the supposed successes achieved by Strauer’s treatment were built on several inconsistencies and practical failings, as British researchers established in several publications. The Heinrich Heine

Photos: MPI for Molecular Biomedicine / Boris Greber; MHH / Mania Ackermann


Berlin-Brandenburg Center for Regenerative Therapies BCRT

The Berlin-Brandenburg Center for Regenerative Therapies (BCRT) is an interdisciplinary translational center with the goal of enhancing endogenous regeneration by cells, biomaterials, and factors which can be used to develop and implement innovative therapies and products. At the BCRT clinicians and researchers are working closely together on a personalized medicine that depends on the early recognition of patients‘ individual healing potential. The primary focus of the BCRT is on diseases of the immune system, the musculoskeletal system, 24

the cardiovascular system and the kidney for which currently only unsatisfactory treatment options are available. Early cooperation with industry, health insurers and regulatory authorities as well as other external partners boosts the chances of exploiting new methods and provides access to flexible financing options. BCRT · Charité – Universitätsmedizin Berlin Augustenburger Platz 1 13353 Berlin www.b-crt.de



University in Düsseldorf and its university hospital have commenced disciplinary proceedings on suspicion of scientific misconduct.

In the first decade of the 21st century, doctors worldwide began to use various types of cells from bone marrow, either injecting them into the damaged parts of the heart or infusing them into the heart via the coronary arteries. A milestone in cardiac cell therapy was the large-scale REPAIR-AMI study, which involved 204 patients and was led by Andreas Zeiher and Stefanie Dimmeler of Goethe University Frankfurt. The researchers examined the results of stem cell treatment in heart attack patients, using rigorous scientific criteria. They isolated blood mononuclear cells (BMC) from the bone marrow of heart attack patients and then re-administered them. In the double-blind, placebo-controlled and randomized trial, the cell therapy was found to be safe and effective. However, the beneficial effects for patients were minor. The pumping efficiency of the heart improved on average by two to three percentage points; in the case of extensive infarction the effect was significantly greater. In relation to the combined hard endpoints of death, repeated myocardial infarction, or readmission to hospital, the results are also highly significant statistically. The Frankfurt-based researchers plan to collect more data to substantiate the

usefulness of this regenerative therapy. Andreas Zeiher and Birgit Aßmus, together with many other German clinicians, are heavily involved in the major European approval-related Phase III trial known as BAMI, in which patients who have suffered acute myocardial infarction are treated with autologous BMCs using coronary intervention techniques. Experts regard the BAMI trial, which is being funded by the EU to the tune of six million Euros, as pointing the way forward for cardiac stem cell therapy. “In 2019 we shall be able to say whether patients actually have longer and better-quality lives with this treatment,” said Zeiher in interview with the GSCN. Heart muscle cells are the perfect cell type

Researchers do not yet fully understand how the administered bone marrow stem cells actually work in the heart. They have moved away from the original idea that they transform into heart muscle cells inside the heart. “The cells obviously work indirectly via paracrine effects – in other words, they release factors and modulate inflammation processes. But a few days after the injection the cells have disappeared,” says Wolfram Zimmermann of the Institute of Pharmacology at University Medical Center Göttingen. The cardiologist believes that the starting material for cardiac cell therapy needs to be extremely well defined in order to make trial results more comparable.

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searchers have already obtained impressive results by using their tissue constructs in rats and mice: when implanted in coronary scar tissue, the cell patches integrated well, the cells actively contracted and even integrated electrically, so that pumping performance increased. Human When human cell patches are used in His Göttingen-based team has spent pluripotent stem cells animals, the transplants still survive and many years working on the production support the functioning of the heart. The of heart-tissue constructs by means of tisheart patches will be tested next in experiments sue engineering. Their method involves embedon large animals. “If the treatment is found to be safe, we ding large numbers of heart muscle cells in a collagen want to start a clinical trial in Göttingen and Stanford in matrix. The resulting patches have been christened “engineered heart muscle.” The 16 cm2 tissue patches each a couple of years. It would be the first clinical trial of artificial contain around 40 million cardiomyocytes. “In recent years heart tissue in humans – with all the challenges that brings,” we have optimized the engineering process so that it oper- says Zimmermann. ates robustly and under GMP conditions and can be transHeart patches also being tested in Hannover ferred to clinical practice,” says Zimmermann. Cells travel across the Atlantic

To produce the heart muscle cells in large quantities, the researchers use human ES and iPS cells. These cells are produced in the US – in the laboratory of Larry Couture at City of Hope, using a technique developed by Joseph Wu of Stanford University. It is a German-Californian cooperation project supported by the Federal Ministry for Education and Research (BMBF) and the California Institute for Regenerative Medicine (CIRM). The cardiomyocytes are shipped to Germany, where the researchers in Göttingen use them to make the tissue constructs. Some of the heart muscle patches are even sent back to California. For Zimmermann this demonstrates how transatlantic cooperation can work: “We can overcome the logistical challenges: the shipped cells survive the journey without problems,” he says. He sees the future of cell therapy as being “allogeneic,” with preparations based on cell material that is not taken from the patient: “Everything else is impractical and prohibitively expensive, especially when a broad medical application such as myocardial infarction therapy is involved.” The re-

The team led by Ulrich Martin is pursuing a similar goal at the REBIRTH Cluster of Excellence at Hannover Medical School. “We want to inject heart muscle cells derived from iPS cells into infarcted hearts or administer them via tissue patches,” says Martin, who is also currently President of the GSCN. Following successful tests in rodents, his team has also started experiments on large animals as a means of testing the cell replacement therapy. They are aiming to conduct the first clinical trial in five years. Martin considers the risk of the transplanted cells degenerating in the body and forming tumors to be small. “The technologies are now sufficiently safe and reliable for us to be able to produce good-quality cells in large quantities.” But he comments that a suitable blood-vessel supply for thicker cell patches is still needed. This is something that the regenerative medicine experts in Hannover are currently working on. These examples of the use of stem cells in cardiac therapy show that the latest generation of cell therapies could find their way into German hospitals in the not-too-distant future.

Text: Philipp Graf

Photo: MHH / Mania Ackermann

For substantial regeneration of infarcted hearts, Zimmermann relies on cells of another type – heart muscle cells. “If you want to replace lost and scarred heart muscle tissue, cardiomyocytes are the only option,” says Zimmermann.

Center for Regenerative Therapies Dresden (CRTD) DFG Research Center and Cluster of Excellence at the TU Dresden

At the DFG Research Center for Regenerative Therapies Dresden (CRTD), Cluster of Excellence at the TU Dresden scientists are seeking to understand the mechanisms of regeneration using model organisms to translate the results to man and to develop novel regenerative therapies for thus far incurable diseases. The center’s major research areas are focused on hematology/ immunology, diabetes, neurodegenerative diseases, bone regeneration and technology development. Currently, eight professors and nine group leaders are working at the CRTD. They are 26

integrated into a network of over 80 member labs at 7 different institutions in Dresden. In addition, 21 partners from industry are supporting the research projects. The synergies in the network allow for a fast translation of results from basic research to clinical applications. DFG Research Center for Regenerative Therapies Dresden – Cluster of Excellence Fetscherstraße 105 01307 Dresden www.crt-dresden.de


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INTERVIEW WITH EGBERT FLORY

Interview with Egbert Flory on the authorization of cell therapies

“Stem cells need flexible rules” Anyone seeking to manufacture stem cell-based medicinal products are obliged to fulfil the authorities’ regulatory requirements when moving into clinical application. It is important to retain an understanding of this situation. If stem cells undergo additional engineering on their way to becoming a finished product, they are classified as Advanced Therapy Medicinal Products (ATMP) within the EU. They will then require a centralized marketing authorization from the European Medicines Agency (EMA). Egbert Flory, a virologist at the Paul Ehrlich Institute (PEI) in Langen, is one of Germany’s leading experts in ATMP regulations. He made major contributions to preparing the EU regulation that came into force at the end of 2008, and continues to serve on the EMA committee responsible for decision-making on ATMPs. At PEI, Flory is the deputy head of the Medical Biotechnology Division. In addition to managing the regulatory work there, he also runs his own laboratory that performs research into stem cells.

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Mr. Flory, what are the current major developments in the field of stem cell-based medicinal products in Europe and in Germany? Ever since the ATMP regulation entered into force in 2008, we have been experiencing a sharp rise in clinical developments and applications for stem cell therapies – particularly those that are based on adult stem cells, such as mesenchymal stem cells (MSCs). Surveys show that just below 200 European clinical studies are focusing on stem cells. I’d estimate that Germany has about 35 such studies. So far, there are no clinical trials or approved ATMPs based on iPS cells in Europe. However, developers have been seeking out scientific advice for this area – including in Germany. Several EU member states currently also host a number of clinical studies that are using human embryonic stem cells as their starting materials. Photo: PEI / Egbert Flory

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Why are stem cell-based products such a big challenge from a regulatory point of view? Stem cells are extremely diverse, so it is difficult to characterize them. Furthermore, the different types of cells have very different risk profiles. This presents major challenges for regulators and the applicants them-


selves. Even the starting material is highly heterogeneous: there are pluripotent stem cells, adult stem cells, and a variety of “precursor cells.” The manufacturing processes are usually very complex, and biological assays are often not established and validated in a pharmaceutical sense. In some cases, only a few relevant animal models are available for the non-clinical development phase. Besides, the individual characteristics of patients also makes things complicated.

In your view, what functions should a regulatory framework for stem cell-based therapies fulfill? As is the case with conventional medicinal products, it is primarily a question of quality, safety, and efficacy. However, for years my mantra has been that we desperately need a flexible regulatory framework for stem cell-based medicinal products. They have to be considered individually, on a case-by-case basis. To achieve this, we need a close, multidisciplinary collaboration between scientists, expert organizations, and regulators. Regulatory decisions regarding stem cell products must be based on scientific knowledge and must take into account the specific risk profile of the cells. The regulatory authorities call stem cell-based products ATMPs. This means they fall within a European legal framework. What are the key points of the EU’s ATMP rules? The EU regulation on ATMPs entered into force in December 2008. This created a harmonized, EU-wide process for the highly complex and innovative cell-based therapies. Essentially, this means that these types of products cannot be placed on the European market without having received approval centralized marketing authorization. The European Medicines Agency is responsible for granting the latter, and reaches its decisions in London. A special EMA committee was also set up – the Committee for Advanced Therapies (CAT) –to take charge of ATMPs as a special group of medicinal products..

Egbert Flory

What exactly is the CAT’s role? The CAT is the body that reviews all the data on a product. Thus, for ATMPs in Europe, it is the supreme committee for preparing a decision on an authorization via

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providing developers with scientific advice, and providing expertise during inspections. The state authorities carry out the pharmaceutical monitoring, including the inspections. They also grant the manufacturing authorizations for ATMPs, based on the European Good Manufacturing Practice guidelines. The state authorities also categorize medicines in accordance with pharmaceutical legislation – they perform the classification, which is legally binding in Germany.

What are the tasks of the Paul Ehrlich Institute (PEI) and the state authorities here in Germany? As a federal authority and national licensing authority for biomedicines, the PEI is legally responsible for all processes relevant to ATMPs – i.e. for authorizing clinical studies,

How many ATMPs have been approved in the EU to date? Six products have been approved in the EU so far, and the process is underway for four others. That is rather a low output.

Photo: MDC

the European Commission. Every member state appoints two experts to the committee. My PEI colleague Dr Martina Schüssler-Lenz and I are currently the members for Germany. The CAT makes the decisions, provides scientific advice, and assigns the cell products according to their properties, i.e. performs the so-called classifications. The committee considers future developments in the ATMP field. We meet at least once a month. The CAT also includes members who represent patients and clinicians. This diversity is rare at the EMA and underscores the committee’s very multidisciplinary approach.

Have there been any notable changes to the ATMP rules in recent years? The regulations are based on the EMA’s Guideline on Human Cell-based Medicinal Products from 2007. This is the parent guideline for all cell-based therapies, and it was a global pioneer. Even back then, the aim was to remain open and flexible. In 2010, the CAT published the world’s first stem cell guideline, which also has a great deal of influence outside Europe. In recent years, the CAT has also focused extensively on the classification of cell-based medicinal products. Stem cell-based products are not only considered as ATMPs when they have been “substantially manipulated” using biotechnological processes; cell products are also classified as ATMPs if they are applied in a “non-homologous” manner – in other words, if the cells have different functions in the donor and the recipient. The CAT published classification recommendations relating to these aspects last year. The observations contained in this “reflection paper” are especially helpful with regard to stem cell products.

Chair Tissue Engineering and Regenerative Medicine (TERM) Artificial Vascularized Tissues

Implants with autologous cells are engineered by Tissue Engineering methods which minimize the body´s own implant rejection. The hereby activated self-healing mechanisms result in tissue regeneration (Regenerative Medicine). We develop human test systems as alternative tissue models to animal trials. Tissue specific bioreactors providing culture conditions of the cell´s natural microenvironment in the body are created to ensure in vitro functionality of the used cells. A biological vascularized scaffold, the BioVaSc, is applied to generate vascularized 30

tissue in vitro. Tissue models as well as models for cancer or (infectious) diseases have successfully been set up. Chair Tissue Engineering and Regenerative Medicine (TERM) Universtiy Hospital Würzburg Röntgenring 11 97070 Würzburg www.term.ukw.de

Tissue-specific bioreactors and incubators.



Stem cell-based ATMPs: The regulatory path to the European market

Scien#fic advice for developers in Germany

Basic research

Pre/Non-clinical development

Na#onal level: scien&fic advice by Paul Ehrlich Ins&tute (PEI)

Phase I/II

European level: Briefing mee&ng EMA - ITF

Discussion of scien+fic principles, development strategies

Photo: MDC / Graphic: PEI / Egbert Flory

ATMP – Advanced-Therapy Medicinal Product ITF – Innova&on Task Force SWAP – Scien&fic advice working party

Phase III

Market Post authoriza&on authoriza&on studies

European level EMA - SAWP

Discussion of scien+fic principles & classifica+on

Discussion of specific ques+ons for market authoriza+on answered by CAT & CHMP

CAT - CommiKee for Advanced Therapies CHMP - CommiKee for Medicinal Products for Human Use

I don’t see it that way. The number is appropriate for a field that is still in its infancy. It is important to remember that we are dealing with very complex, innovative, and scientifically oriented products. Not so many of those exist. And the number of applications is rising steadily. Authorizations in this field are just as rare in the U.S. and Japan as they are here in Europe.

There was of course an important premiere in April 2015, when Holoclar became the first stem cell therapy to be granted a marketing authorization in the EU. How does this product work? Holoclar is a stem cell treatment used in the eye. It is intended for patients who have suffered damage to the surface of the cornea, for instance as a result of physical or chemical burns. Limbal stem cells are removed from the edge of the patient’s cornea, cultivated in the laboratory, and then re-transplanted into the damaged areas. An Italian company manufactures the product. The clinical

data were convincing, with many patients experiencing improvements to their eyesight. The product was granted what is known as a conditional authorization. This means that additional confirmatory clinical studies are being run in several centers, including in Germany. The developers thus have to submit further data on the efficacy of the medicinal product. What went well with Holoclar? What can future developers learn from it? It was the fastest ATMP process that my team and I have handled so far. Once the application had been submitted, it took just one year to reach a decision. The manufacturer in Italy was very well connected and organized. Another plus point was that the scientific development work, which had lasted many years, was extremely good and transparent. Overall, that is the ideal basis for our regulatory work. It means we can still work if there are only limited data from non-clinical studies or if no randomized clinical trials have

Fraunhofer Research Institution for Marine Biotechnology EMB Innovative research & development at the new institute building of the Fraunhofer EMB in Lübeck

For one year the Fraunhofer EMB is working now in the new institute building on the Lübeck BioMedTec Science Campus. The newly constructed building has got modernly equipped laboratories, aquaculture facilities, a food technology center and biobanks with fully automated state-of-theart technology covering a total area of 8 292 m² (BGF).The technical re-equipment includes a X-ray microscope, a non-invasive small animal MRI as well as several 3D printer of the latest generation, which are used for the development of novel laboratory appliances. Annual GSCN Magazine 2015/16

“With these excellent research capacities we look forward to strengthening the life science expertise of the Fraunhofer-Gesellschaft. The new research building gives us the opportunity to explore promising topics for applied research and to promote existing business areas with the most modern equipment” concludes Prof. Charli Kruse, director of the Fraunhofer EMB. The Fraunhofer EMB works on industry-related research topics with focus on life sciences. Here, novel technologies, procedures and instruments for cell isolation and exploitation were developed. Moreover, the scientists from the

EMB work on innovative aquaculture systems and on the utilization of aquatic raw materials for food engineering. With the “Cryo-Brehm” the EMB maintains one of the largest archives for cell cultures from wild animals. Fraunhofer-Einrichtung für Marine Biotechnologie EMB Mönkhofer Weg 239a 23562 Lübeck www.emb.fraunhofer.de

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been carried out. We are gaining many valuable experiences from the first approved stem cell product. This will certainly facilitate the process for subsequent applicants.

At the Paul Ehrlich Institute, you offer scientific advice to medicinal product developers. How does this work? Since 2009, the PEI Innovation Office has been providing interested parties with advice on ATMP development at any time. Fortunately, many developers have shed their initial hesitance to make contact. We conduct about 50 advisory sessions a year. These might be intensive consultations similar to those held at the EMA, or they could be orientation discussions, which involve an informal chat without any major documentation. Seeking advice makes particular sense in the early phase of product development and in the run-up to clinical studies, as it can help developers avoid potential errors that will require a lot of extra work and money at a later stage. The costs are also attractive: the hourly rate per expert is just € 68.

After a marketing authorization has been granted, companies have to start grappling with the problem of cost reimbursement at a national level. Unlike the authorizations, cost reimbursement is not organized centrally in Europe. Each country therefore decides how the costs of a therapy are to be reimbursed and who is responsible for it. We have comparatively generous reimbursements for cell-based therapies here in Germany. The situation is very different in other EU countries. That

can be frustrating for manufacturers. Take, for instance, Provenge, a tumor vaccine for prostate cancer that was authorized as an ATMP in 2013. The manufacturers of this immunotherapy have since withdrawn the EU marketing authorization because the future reimbursement situation in Europe was unresolved. Germany was the only EU country where health insurance providers reimbursed the costs. The EMA and PEI are now working very hard to include reimbursers in the scientific advisory sessions at an early stage – i.e. before the authorization process begins.

What does the term “hospital exemption” mean? How does the rule affect developers of stem cell-based therapies in Germany? The exemption, which is anchored in the ATMP regulation, exists because a great deal of innovation happens in hospitals, and because local production and application are very common. The European Commission formulated the exemption in order to allow these non-routine applications to be used in hospitals in a very limited way. EU countries have taken quite different approaches to implementing it. Germany has implemented it in a very interesting manner, I think: once innovative products receive PEI authorization, hospitals can use them on patients to a limited extent. The manufacturing and processing can be done by a toll manufacturer.

Photo: MDC

When it comes to developing a stem cell-based medicinal product, how important is the classification? Classification first involves defining which medicinal track companies or universities are involved in their product. I think it is important that developers know this at an early stage. Seeking a recommendation for a classification is often the first contact that an applicant has with the relevant authority. However, an EMA classification – which takes about six weeks – is not a legally binding scientific recommendation. In Germany, the state authorities are responsible for legally classifying medicinal products. It is not an official task of the PEI, but since we obviously have specialist expertise in the field, the state authorities generally ask us for advice.

The Collaborative Research Center SFB 873 Maintenance and Differentiation of Stem Cells

The Collaborative Research Center SFB 873 “Maintenance and Differentiation of Stem Cells in Development and Disease” at Heidelberg University works towards defining the regulatory principles underlying the balance between maintenance, expansion and differentiation of stem cells in diverse systems on a mechanistic level. To this end the SFB873 studies a wide spectrum of experimental models ranging from plants to human to elucidate the inherent properties of specific stem cell systems, but also to uncover common and divergent principles behind regulatory regimes and molecular signatures. 32

Our consortium brings together internationally recognized researchers, with unique scientific strengths in cell biology, biophysics, developmental biology, molecular medicine or modeling. With our research we hope to advance our understanding of principles underlying stem cell function and lay the foundation for translational approaches. Centre for Organismal Studies (COS) Im Neuenheimer Feld 230 69120 Heidelberg www.sfb873.de



The manufacturer has to be in Germany but does not have to be located in the hospital. This creates interesting possibilities for specialized companies. In Germany, seven developers have so far been granted a national exemption under Section 4b of the country’s Drug Law. Nevertheless, our feeling is that the hospital exemption is a step along the way toward a central marketing authorization – and should remain an exception. Interview: Philipp Graf

Suggested reading

Egbert Flory, Paolo Gasparini et al. (2015), “Regulatory viewpoints on the development of advanced stem cell–based medicinal products in light of the first EU-approved stem cell product”, Cell & Gene Therapy Insights, DOI:10.18609.cgti.2015.010

What are ATMPs? Cell-based medicinal products that contain living or non-living cells and were engineered during manufacturing are legally classified as Advanced Therapy Medicinal Products (ATMP) in the EU. The European Medicines Agency (EMA) is responsible for the marketing authorization of these products. ATMPs are divided into different classes: gene-therapy medicines, tissue-engineered products, somatic-cell-therapy medicines, and combinations thereof. Six products have been granted a marketing authorization in the EU since the ATMP regulation entered into force: Product name Manufacturer Year of EU marketing authorization ChondroCelect TiGenix 2009 Glybera UniQure 2012 Provenge Dendreon 2013 1 MACI Genzyme 2013 2 Holoclar Holostem Advanced Therapies 2015 Imlygic Amgen 2016 1

Since withdrawn at manufacturer’s request

2

Suspended due to manufacturing stoppage

Disclaimer: Please take notice, that the online version of the interview is slightly different to the printed version due to further editing.

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STEM CELL-BASED DISEASE MODELS

Stem cell-based disease models

Organoids: 3D miniatures are booming “Organoids” are as tiny as mustard seeds, but many biomedical scientists believe they are the next big trend in health research. Using sophisticated 3D tissue culture techniques, researchers are becoming increasingly skilled at combining stem cells and differentiated cells to create miniature versions of organs such as brains, intestines, and stomachs. The topic is also attracting more and more stem cell researchers in Germany. Organoids are so similar to the full-sized organs on which they are modeled that they have opened up new avenues for investigating principles of developmental biology and disease etiologies. This means that the mini-organs are not only good candidates for testing active substances, they also have enormous potential for diagnostics and regenerative therapies.

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ina Bartfeld is delighted every time she looks at her mini-stomachs, just days old, through the microscope: “My organoids are so beautiful,” says Bartfeld, a biologist who is currently setting up her own group at the University of Würzburg. The delicate, hollow spheres are made of a single layer of cells and measure just under half a millimeter. Smaller bubbles are budding around their edges. “As well as various differentiated cell types, these also contain the stem cells that constantly replenish the stock of cells,” says Bartfeld. It takes less than two weeks for a tiny stomach to grow. “The fascinating thing is that the stomach organoids self-organize and keep growing,” says Bartfeld.

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Photo: Helmholtz Zentrum München / Marisa Karow

Bartfeld is one of a rapidly increasing number of scientists whose work with organoids is changing biomedical research. From miniscule intestines and lungs to diminutive mammary glands and brains, tiny versions of almost every organ in the human body are flourishing in laboratories all over the world. As well as resembling the full-sized organs in appearance, the in vitro organoids also carry out astoundingly realistic complex biological processes.

Bartfeld moved to Würzburg from Utrecht in the Netherlands last fall. As a postdoc, she worked at the Hubrecht Institute for Developmental Biology, in the working group of Hans Clevers, a Dutch stem cell researcher who is considered the pioneer and initiator of the recent cell-culture boom. From the elite training center of the Hubrecht Institute, organoid technology is now conquering research laboratories all over the world. Adult stem cells back in the spotlight

Three-dimensional cell cultures are not a new trend. Researchers involved in the differentiation of pluripotent stem cells are familiar with “embryoid bodies,” three-dimensional clumps in which the cells develop much faster than they would in the body. Tissue engineers have also spent many years experimenting with 3D aggregates made from different types of cells.


“Hans Clevers’ method is revolutionary because it allows us to isolate adult stem cells from organs and to recreate their niche in the Petri dish,” explains Bartfeld. Clevers and Toshiro Sato first accomplished the feat in 2009, with tissue from the small intestine. One of the keys to the team’s success was that they had identified an important stem cell Cerebral organoids surface marker (the protein LGR-5) that made it easy to isolate adult stem cells from intestinal tissue removed during surgery. The marker can also be used to find adult stem cells in other tissues. Sophisticated cell-culture techniques paved the way for another step forward: by adding Matrigel, a gel-like secretion from mouse tumor cells, the scientists made a very good version of the immediate environment (“niche”) of adult stem cells. Once they are embedded in this extracellular matrix, the cells find themselves in very comfortable surroundings. Adding growth factors and a few other types of cells was then enough to make the adult stem cells do in the Petri dish what they would do in an actual intestinal wall: renew themselves while simultaneously producing structures like villi and crypts. As a result, a self-organizing, living intestinal system grew in the Petri dish. Realistic model systems for diseases

“This culture technique provides an inexhaustible source of human cells from a specific type of tissue,” says Bartfeld. “And what is more, the cells are unchanged.” Bartfeld explains that this means the organoids get much closer to the natural, in vivo situation than cell aggregates, which are obtained using pluripotent stem cells. Another advantage is that it is easy to learn how to obtain the cells and perform the culture technique. The mini-organs are also easy to handle. “Freezing, defrosting, mailing – none of that is a problem,” says Bartfeld. Bartfeld explains that the field exploded after Nature published the discoveries in 2009. Adult stem cells, which had retreated into the background in the era of iPS cells, returned to the spotlight. From then on, many of Bartfeld’s colleagues in Utrecht began picking out their own organ system. As cell-based model systems, organoids are not only well suited to studying organogenesis and the developmental biology of tissues. They also open up entirely new scope for investigating disease mechanisms. At the Institute for Molecular Infection Biology in Würzburg, Bartfeld is hoping to

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use her mini-stomachs to recreate the development of peptic ulcers. To do so, she is infecting the 3D structures with Helicobacter pylori bacteria and observing closely how the germs cause cancer to develop. “This means we can simulate and examine in more detail the interaction between the pathogen and the human host cells.” Bartfeld’s initial experiments have already shown that in organoids infected with the stomach bacteria, key steps involved in the innate immune response occur correctly, although the pattern varies depending on the type of cell.

Mammary gland organoide grown in a petri dish

Henner Farin, who since September 2015 has been running a young researchers’ group at the Georg Speyer Haus in Frankfurt as part of the German Consortium for Translational Cancer Research (DKTK), is another protégé to come out of Clevers’ laboratory and return to Germany. Farin plans to produce intestinal tumor organoids and use them to research the mechanisms of carcinogenesis. He is also planning to use patient-specific organoids to investigate chronic inflammatory intestinal diseases. News from the organoid workshops

Another reason why health researchers are attracted to working with organoids is that they are easy to combine with other cell technologies. For instance, scientists can equip them with cell types derived from induced pluripotent stem cells (iPS cells). Individual cells in the miniature organs can also be modified or reprogramed using genome editing. This means the 3D structures can be tailored for a specific disease model. The GSCN annual conference in September 2015 showcased what researchers working on 3D cell cultures are now capa-

ble of. Hans-Willem Snoeck of Columbia University in New York reported on his attempts to recreate the development of a human lung as naturally as possible in the Petri dish. With the help of iPS cells and a sophisticated protocol, his team has made remarkable progress. “We have succeeded for the first time in allowing the fine branching and the maturation of the alveoli to occur in the Petri dish,” says Snoeck. In addition to producing insights into developmental biology, the models can also be used to study numerous lung diseases, such as the flu, cystic fibrosis, and lung cancer. A team led by Christina Scheel at Helmholtz Zentrum München, meanwhile, has succeeded in recreating the complex 3D structures of the mammary glands. The Munich researchers isolated stem cell-like cells from tissue removed during cosmetic breast-reduction surgery. In the 3D culture, the cells begin forming complex milk ducts that end in bubble-like structures. The researchers now want to use the mini-breasts to examine the development of breast cancer in more detail.

Photo: Helmholtz Zentrum München / Jelena Linnemann, Lisa Meixner

Another potential application is in regenerative medicine. For instance, Bartfeld has succeeded in making mini-stomachs from patients who have had their stomachs removed to treat cancer. “One use for these cancer organoids is to test which medicines are effective and which are not,” says Bartfeld. She also explains that the organoids can be used to simultaneously cultivate and compare healthy and diseased tissue from the same patient.

Institute for Stem Cell Research and Regenerative Medicine (ISRM) Regenerative Medizin at a glance

At the institute for stem cell research and regenerative medicine we adopt a systems biology approach to better understand normal development (hepatogenesis and neurogenesis), ageing and disease mechanisms (Steatosis/Non Alcoholic Fatty Liver Disease, Alzheimer’s disease, Nijmegen Breakage Syndrome and Crigler-Najjar syndrome). We are actively involved in the EU/ FP7 funded project AgedBrainSYSBIO (http://agedbrainsysbio.eu) where we study mechanisms underlying late onset Alzheimer’s Disease (LOAD).Mechanisms underlying the induction and maintenance of pluripotency and 36

the derivation of patient specific induced pluripotent stem cells (iPSCs) differentiated into relevant cells types (2D and 3D) are core to these efforts. Omics-based datasets (transcriptome, proteome, methylome and secretome) Bioinformatics, mathematically modeling, pathway reconstruction and data management are central to our research. Heinrich-Heine-Universität Universitätsklinikum Düsseldorf · ISRM Moorenstr. 5 · Geb. 14.80.01.002 · 40225 Düsseldorf www.uniklinik-duesseldorf.de/ISRM



Reprograming mini-brains No organoid has caused as much of a stir over recent years as the pea-sized mini-brains that have been grown in Jürgen Knoblich’s laboratory at the Institute of Molecular Biotechnology in Vienna. Neural precursor cells derived from iPS cells began, almost of their own accord, combining to form so-called cerebral organoids. The little balls of neural tissue resemble, both outwardly and in terms of their molecular biology, the structures of a young embryonic brain. “In 2013, this work was definitely an eye-opener for the field and created a great sense of optimism,” says Benedikt Berninger of the Institute for Physiological Chemistry at the University of Mainz. His working group is one of many in Germany that have begun focusing on cerebral organoids. “We use the mini-brains as a platform for studying the reprograming of glial cells to form neurons,” explains Berninger. His colleague Marisa Karow carries out cell transplants on the tiny brains – a kind of open brain surgery in the Petri dish. It is then possible to follow the consequences of this intervention in real time. The mini-brains have also given the neuroscientists new avenues for deciphering the molecular programs that occur in developmental disorders in the brain.

Berninger believes that the current wave of euphoria surrounding the organoids is absolutely justified, but adds that “we are still in the early phases.” He says that the research is expensive and complex, and that key questions relating to the long-term survival of the cell cultures have yet to be answered. The cell-culture experts are still trying to work out how best to look after and feed the organoids over a period of many months. Berninger is campaigning for better networking of the knowledge and expertise within the growing research community in Germany. Julia Ladewig in Bonn, Frank Edenhofer in Würzburg, and Magdalena Götz in Munich are just some of the stem cell researchers whose teams are working intensively on the living 3D brain models. “We, too, are enthusiastically following and participating in the progress being made with organoids,” says Hans Schöler, Director of the Max Planck Institute for Molecular Biomedicine in Münster. The Max Planck researchers are using brain organoids as test objects for the targeted reprograming of astrocytes to form precursor cells. “We hope that this will improve our understanding of neurodegenerative ageing processes,” says Schöler. “Right now, organoids are a vast playground.” Text: Philipp Graf

New kits on the block… „Pure, what do you mean, pure?!?!?“ Albert grumbled. “It’s not so easy to make pure Dopaminergic Neurons for Parkinson’s disease research. I always see a certain degree of “contamination” with fore- and hindbrain Dopaminergic Neurons during differentiation from iPSCs and ESCs. For example, I just recently used a kit, which was easy to use and gave me high yields of NSCs from my iPSCs. However when further differentiating these NSCs, I achieved not only midbrain DA Neurons, which Parkinson’s researchers are mostly interested in, but I also noticed the presence of a significant number of fore- and hindbrain DA neurons. In this case, the ease-ofuse advantage actually turned out to be a disadvantage for this specific application.” “Well, darling, that is interesting to hear, but how can I be of help in this case?” asked Josephine, the lovely wife of Albert, who didn‘t have a clue about research. “Dad” said Madeline their daughter, a Ph.D. student in her second year, as she joined the discussion, “I just heard about a new kit, which might fit perfectly with your work. With this new kit you can differentiate midbrain dopaminergic neurons from iPSCs and ESCs. That is what your Parkinson’s researchers are looking for, right? I even saw some data on the new kit, which provides a simple to use and reproducible protocol, and is even quicker to use than most of the protocols in common use today. You only need 38 days to get to generate your desired mid-brain dopaminergic cell type, which is 7 days shorter than the 45 days required by most protocols. But much more importantly”, said Madeleine as she started to become euphoric, “you can make a break in between! Yes, you can efficiently expand and bank your progenitor cells! This allows you to start your differentiations from an intermediate stage next time around using the same batch cells and avoids you having to go all the way back to the iPSC or ESC stage!” Annual GSCN Magazine 2015/16

“Wow, that sounds interesting!” said Alfred, interrupting Madeleine, “Thanks for the tip! Do you recall which company makes this kit?” “That’s the new kit from Thermo Fisher Scientific and it is called PSC Dopaminergic Neuron Differentiation kit or something like that. Just yesterday I received the link from one of my lab mates: www.thermofisher.com/de/de/home/life-science/stem-cell-research/stem-celldifferentiation/psc-dopaminergic-neuron-differentiation.html

Just check out the homepage! Oh, sorry, but I have to leave now!” and Madeleine ran off in a hurry as The Big Bang Theory had just started on TV. 37


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Floating cystic micro spheroids of heart muscle cells 38

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STEM CELL-BASED DRUG SCREENING

Stem cell-based drug screening

On the hunt for new drugs

Reprogramming, genome editing and robotics: a mixture of powerful stem cell technologies is fundamentally changing the way drugs are developed. After years of taking a “wait and see” stance, the pharmaceutical industry is now making more and more use of cell-based disease models to accelerate the search for new drugs and improve the chances of success. The first substances identified with the help of pluripotent stem cells are already being tested in clinical trials. New translation centers are now implementing findings from German laboratories in stem cell-based drug screening. And two major European projects are working together to set up a giant stem cell bank.

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t is less than ten years since Japanese scientist Shinya Yamanaka first converted somatic cells into induced pluripotent stem cells (iPS cells). Even then, Yamanaka – who was subsequently awarded the Nobel Prize – and many other researchers were convinced that the first major application of the revolutionary reprogramming technique would be drugs based on cell models.

For pharmaceutical researchers, the refinement of the iPS technique opened up a promising new way of searching for and testing new drugs. In previous years, the pharmaceutical industry had suffered too many costly setbacks. Drugs that had appeared promising failed in the late stages of clinical development, often because the results of experiments with immortalized cell lines or on animals proved insufficiently transferable to patients.

Photo: MDC / Chen Chen

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The iPS technique gives biomedical experts direct access to “authentic” human cell material that they have never had before. In addition, patient-specific cells or even simple miniature organs can be produced, enabling diseases to be modelled in the Petri dish. New tools from molecular biology – in particular genome editing – have vastly increased the options open to stem cell researchers. With designer nucleases such as the now popular CRISPR/Cas9 system, the stem cells’ genetic material can be modified in precise ways. This means that the cells of the disease models can be genetically tailored in the laboratory. The possible functions of gene variants that have already been linked to the emergence of disease through genome analysis can now be systematically examined in a cell-based model in the Petri dish. Producing corresponding control cells further increases the informative value of the results. A new era in pharmaceutical research

In the early days, pharmaceutical research tended to focus on stem cell-based toxicity tests; now, however, drug devel-


opment is also gaining in importance. Worldwide, the first clinical studies are getting under way on substances that have emerged from stem cell-based screening. The major pharmaceutical companies have their sights set on neurodegenerative and psychiatric disorders in particular: • GlaxoSmithKline has started a Phase II clinical trial of the drug Retigabine involving 192 patients with amyotrophic lateral sclerosis (ALS). IPS-based disease models have shown that the drug helps reduce the excitability of damaged motor neurons. • Roche is testing the drug known as RG7800 on patients with spinal muscular atrophy (SMA) in a Phase II trial; the substance has been validated in an iPS-based model. • Bristol-Myers Squibb (BMS) has commenced two Phase I trials of a tau-specific antibody as a treatment for Alzheimer’s. The drug BMS-986168 originated with iPierian, a start-up acquired by BMS in 2014. • Novartis is using drug screening on iPS-based cell models to search for treatments for autism; according to media reports it is planning a clinical trial with a promising candidate. New translation centers set up

In Germany, too, stem cell-based drug screening has gathered pace. “We can no longer complain about a lack of interest from the pharmaceutical industry,” says Oliver Brüstle of the Institute of Reconstructive Neurobiology in Bonn. “Five years ago the situation was completely different: then, the pharmaceutical sector was very hesitant about our ideas, but that has now changed radically.” The shift is reflected in newly launched translation projects with partners in science and industry. The researchers in Bonn, for example, have

Cell types differentiated from stem cells

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From stem cells to drug screening (example for cystic fibrosis mutations)

ropean character of the alliance that makes it an interesting concept. “It promotes direct collaboration between applied stem cell research and the pharmaceutical industry and thus speeds up drug development,” he says. The researchers in Bonn will produce neural cells for various screening projects and send them to Strasbourg.

The collaboration came about partly because Brüstle’s team at the Bonn translation center LIFE & BRAIN turned their attention at an early stage to the issues of standardization and automation that are crucial for the pharmaceutical industry. “This involved lengthy and sometimes tedious work,” says Brüstle. “But it enabled us to find a common language.” Another flagship automation project is the StemCellFactory. This robotic production line built at LIFE & BRAIN with support from the state of North Rhine-Westphalia is a fully automated system for producing iPS cells that will also manage their maturation into neural cells. The system is intended for use in large-scale centers and cell banks; biobanks have already signaled their interest in the machine. “The StemCellFactory is currently a prototype and not yet in series production. Further improvements to the system are under way, and we want to incorporate other components such as genome editing,” says Brüstle.

Graphic: MHH / Ulrich Martin; Photo: MDC / Jochen Meier

forged an alliance with the Franco-German translation center KSILINK in Strasbourg: using artificially produced brain cells they will test potential drugs for the treatment of neurodegenerative diseases. KSILINK is a public-private partnership funded by the French government and pharmaceutical company Sanofi. “The aim is for KSILINK to become a bi-national platform for stem cell-based drug development,” says Brüstle. In his view, it is not only the Eu-

Institute of Reconstructive Neurobiology From disease modeling to stem cell therapies The Institute of Reconstructive Neurobiology at the University of Bonn Medical Centre focuses on the use of pluripotent stem cells for the study and treatment of neurological disorders. Based on a broad technology portfolio including cell reprogramming, neural differentiation, direct cell fate conversion, stem cell industrialization and neurotransplantation, the Institute develops stem cell-based model systems for disease-related research and drug development as well as novel cell therapy regimens. It closely 40

interacts with LIFE & BRAIN GmbH, a translational hub of the University of Bonn providing stem cell products and services for pharma, biotech and academia. Institute of Reconstructive Neurobiology LIFE & BRAIN GmbH University of Bonn Sigmund-Freud-Straße 25 53105 Bonn www.stemcells.uni-bonn.de



CARE comes to Munich Another publicly funded translation center for drug development based on iPS cells is to be set up in Munich – the Center for Advanced Regenerative Engineering, or CARE. The Bavarian state parliament approved start-up funding for the center in late 2015. An initial sum of €15 million will be pumped into the development of CARE over the next three years, “CARE is likely to be set up on the Biotech Campus in Martinsried,” says Schöler. The center is due to start work in January 2017 with the aim of using iPS cell technology and patient-specific disease models to develop high-throughput assay formats for drug research. “We plan to focus mainly on neurodegenerative diseases. In the search for new therapeutic approaches we are in principle prepared to investigate all types of cells and diseases,” says Schöler. CARE is to become a center of excellence for regenerative medicine – a place where drug research companies of all sizes from large to small can get actively involved. “Many companies have already announced their interest,” reports Schöler. Science Manager Ulrich Gerth will use 2016 to set up the institute and recruit the first cooperation partners. CARE will strengthen the existing biomedical regional networks in Bavaria, which include m4, the top-level cluster for personalized medicine supported by the German Ministry of Education and Research (BMBF), and forIPS, the Bavarian Research Network Induced Pluripotent Stem Cells. Through these efforts, CARE will also enhance international visibility.

Photo: CRTD / Ruslan Rust

New drugs for cystic fibrosis

The EU is also supporting translational approaches to stem cell-based drug screening – for example in the ERA-Net initiative E-rare, in which European research networks are developing treatments for rare diseases. One of these projects, INSTINCT, was launched at the end of 2015 and is being coordinated by Ulrich Martin of the Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO) at Hannover Medical School (MHH). “We are looking for new drugs to treat cystic fibrosis using patient-specific

Immunohistochemical staining of an adult mouse retina section

iPS cell lines,” says Martin, who is also the current GSCN President. Cystic fibrosis is a metabolic disorder caused by a defect in the genetic blueprint for the ion channel regulator CFTR. Although the first drugs have recently come on the market, only five percent of patients are benefitting from this causal therapy. Their patient-specific stem cells are being matured into epithelial lung cells in the Petri dish by Martin’s team of researchers. “We have produced cell lines with mutations in the CFTR gene that had not previously been widely studied. Now that we have reporter

Heidelberg Institute for Stem Cell Technology and Experimental Medicine HI-STEM gGmbH

HI-STEM gGmbH is a non-profit public-private partnership between the German Cancer Research Center (DKFZ) and the Dietmar Hopp Foundation (DHS).

Located within the DKFZ in Heidelberg, HI-STEM performs cutting-edge research on stem cells with the aim of translating these results into novel clinical applications. This includes the development of novel diagnostic tools and innovative therapies to monitor and target leukemic and solid tumor stem cells as well as metastatic disease. Annual GSCN Magazine 2015/16

Professor Dr. Andreas Trumpp and four Junior Group Leaders direct an international research team of more than fifty employees. The HI-STEM Research Groups: • Hematopoietic and Leukemic Stem Cells (A. Trumpp) • Experimental Hematology (M. Milsom) • Stress induced activation of HSCs (M. Essers) • Cancer Stem Cells and Metastasis (A. Trumpp & M. Spick) • Metastatic Niches (T. Oskarsson)

HI-STEM gGmbH German Cancer Research Center (DKFZ) Im Neuenheimer Feld 280 69120 Heidelberg www.hi-stem.de

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genes we can carry out high-throughput screening of drug libraries.” The research partners are a very international group, with representatives from Italy, Portugal, the Netherlands and Canada. Interesting substances will be taken as far as the pre-clinical phase of drug development. Two IMI consortia on iPS cells

Stem cell-based techniques not only help in the identification of new drugs: In pharmaceutical research, they have also become important tools in the lead validation and optimization of drug candidates found through conventional means.

of pharma and toxicology

The EBiSC consortium coordinated by the pharmaceutical corporation Pfizer has been working since 2014 on produc-

Dentate gyrus (part of the brain structure hippocampus) infected with the human immunodeficiency virus (HIV)

ing well-characterized iPS cell lines and systematically archiving them in a non-commercial bank. The central biobank is located in Cambridge, UK. A mirror bank that will store a complete equivalent of the EBiSC collection is being created in Sulzbach in the Saarland region of Germany. EBiSC, which has a budget of €35 million, is a consortium of 26 organizations, including seven German partners. The StemBANCC consortium, with a budget of €55 million, was launched in 2012. It is coordinated by Swiss pharmaceutical company Roche. Nine of the 35 partners are from Germany. The consortium partners have an ambitious target: to collect skin samples from 500 patients and to generate from them 1,500 cell lines that will then also be included in the EBiSC collection. The focus is on neuronal and neurodegenerative disorders and diabetes.

drug sc

The iPS technology has now acquired an established place in the laboratories of the pharmaceutical industry. Stem cells have become a versatile tool for drug developers, and if the new drug candidates perform well in clinical trials they will play a major role in the medicine of the future.

Fraunhofer Institute for Molecular Biology and applied Ecology IME The Fraunhofer IME conducts research in applied life sciences from a molecular level to entire ecosystems. Our interdisciplinary organization and laboratories with most recent equipment including GMP facilities and complex facilities for environmental simulations allow a wide spectrum of research and development services. IME’s overarching goal is the development and use of novel technologies for diagnosis and therapy of human and animal disease as well as protection of crop plants and food supplies. IME has close ties in terms of personnel and areas of work with the Institute of Molecular Biotechno42

Photos: CRTD / Sara Bragado Alonso, Lara Marone & Ruslan Rust

To do this, pharmaceutical researchers need cells that are produced under standardized conditions, quality checked, catalogued, and available in large quantities. Central European resources are currently being developed in two large consortia as part of the Innovative Medicines Initiative (IMI). IMI is a public-private partnership between the European Commission and the European Federation of Pharmaceutical Industries and Associations (EFPIA). Stem Cells as tools: Development

logy of the RWTH Aachen University, the Department of Biology and Biotechnology of Plants of the University Münster, the Department of Applied Entomology of the University Gießen and the Institute for Clinical Pharmacology of the Goethe University Frankfurt/ Main.

ste c

Text: Philipp Graf

IME Locations: Schmallenberg, Aachen, Münster, Gießen, Frankfurt/M., Newark/Delaware (USA) and Santiago de Chile. Employees: about 680.

Fraunhofer IME Forckenbeckstraße 6, 52074 Aachen Fraunhofer IME ScreeningPort Schnackenburgallee 114, 22525 Hamburg www.ime.fraunhofer.de




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PRECLINICAL STEM CELL RESEARCH

Preclinical stem cell research

Good models needed Before stem cell-based therapies can be tested on patients, they must first be extensively investigated in a preclinical development phase. Not all processes in the body can be simulated in the test tube or on the computer, so experiments on suitable animal models are essential. But stem cell technologies can also help

to reduce the need for animal experiments in drug development. Cell cultures produced using pluripotent stem cells enable toxicology tests to be performed at an early stage, and multi-organ chips may soon make it possible to investigate the human body in miniature format.

Photo: MDC / Maciej Czajkowski

Stained facial muscles of a mouse

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A

ny attempt to use cells as medical products involves dealing with highly complex, living biological systems – which makes working with them a special challenge. Before any clinical application can be trialed on humans, the safety profile and mode of action of the cell therapies must be thoroughly tested in living organisms. Animal experiments are controversial, but for the foreseeable future they will remain indispensable for biomedical research and application, because not all processes in the body can be simulated in the test tube or on the computer. Innovative cell therapies differ fundamentally from conventional pharmaceutical substances in terms of their pharmacology and toxicology. This means that developers of cellbased medical products must draw up individual concepts for their products in order to provide the authorities with convincing data.

Photo: TU Berlin_TissUse / Reyk Horland

Small-animal models are of limited usefulness

But which animal model should be used to obtain the most informative results? The regulatory authorities favor what are known as homologous animal models. This means that the therapeutic effect of cells of a certain species is researched in a corresponding disease model – such as rat cells in rats with myocardial infarction. In other cases, heterologous models can be used – for example if human cell types are transplanted into rodents with a suppressed immune system or into “humanized” animals in order to prevent rejection. Small-animal models – especially rodents such as rats and mice – are used in particular for early feasibility studies. “But in the later stages of development the use of appropriate large-animal models such as sheep or miniature pigs is unavoidable,” says Thomas Braun, Director of the Max Planck Institute for Heart and Lung Research in Bad Nauheim. Braun, a developmental biologist, works exclusively in basic biomedical research; he studies muscle stem cells and the regeneration of heart muscle. Mice are some of his most important subjects

This Four-Organ-Chip contains an intestine, a liver, a kidney and a skin module.

New requirements bring extra bureaucracy According to figures from the Federal Ministry of Agriculture, 2.8 million animals were used in scientific studies in 2014 – 870,000 of them in basic research and 333,000 in translational research. While the overall number of laboratory animals has fallen slightly, the number of transgenic rodents continues to rise – a trend that is ascribed to the steadily growing use of genetic engineering.

As a result of the new EU directive on the protection of animals used for scientific purposes, Germany’s animal protection law has been amended. The new legislation came into force in 2013, with notable consequences. “The work involved in obtaining approval for animal experiments has increased dramatically,” says Braun. “Regardless of whether we are using mice or zebra fish, we must now assess each individual transgenic stem for possible stress and in some cases obtain approval for an animal experiment from the authorities.” Since breeding animals are also covered by this assessment, the overall documentation requirements

German Cancer Research Center (DKFZ) Research for a life without cancer

The German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), located in Heidelberg, is a member of the Helmholtz- Association and is the largest biomedical research institute in Germany. At the DKFZ, more than 1,000 scientists work together in order to develop novel strategies aimed at improving the prevention, diagnosis and treatment of cancer. Several research laboratories investigate normal and cancer stem cells (CSCs) as well as their respective niches. The goal is to develop strategies to monitor and Annual GSCN Magazine 2015/16

target CSCs in primary cancers and metastasis. Together with the Dietmar Hopp Foundation, the DKFZ is a shareholder of HI-STEM, the nonprofit Heidelberg Stem Cell Institute and organizes the biannual Heinrich-Behr-Conference on „Stem Cells and Cancer“, which attracts international experts in the field.

With the Heidelberg University Clinic, the DKFZ has established the National Center for Tumor Diseases (NCT), to clinically translate innovative basic cancer and stem cell research discoveries into clinical therapies.

German Cancer Research Center Im Neuenheimer Feld 280 · 69120 Heidelberg www.dkfz.de

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Alternatives to animal experiments

Although it might be desirable from an ethical point of view to replace all animal experiments with alternative methods of drug development, experts regard this as highly unfeasible in the foreseeable future. Nevertheless, in vitro models and simulations in silico are becoming increasingly informative and realistic; in some tests for the cosmetics industry they have already completely replaced animal experiments. Cell cultures enable toxicology studies to be carried out on potential drug candidates, especially when this involves testing their effect on heart and liver tissue in the Petri dish. Cardiotoxicity and hepatotoxicity are among the most common reasons why potentially promising substances are ruled out for further drug development. Researchers investigating alternative methods use the principle of the 3Rs, developed in the 1950s. The 3Rs stipulate

Cerebral cells: astrocytes

that the methods used should either avoid or replace the use of animals (replacement), minimize the number of animals used in experiments (reduction), or minimize animal suffering (refinement). The German Centre for the Protection of Laboratory Animals (Bf3R) was established in Berlin in 2015. It forms part of the Federal Institute for Risk Assessment (BfR) and is funded by the Federal Ministry of Agriculture. Bf3R focuses on methods that completely replace animal experiments – including stem cell-based cell culture tests such as the embryonic stem cell test. This uses mouse ES cells that are differentiated into heart muscle cells, nerve cells or bone cells. These cells can then be tested to discover whether substances might be toxic to embryonic development. Elsewhere, too, scientists are refining stem cell-based methods for use in pharmacology and toxicology research. Pharmacologists led by Thomas Eschenhagen at the University Medical Center Hamburg-Eppendorf (UKE) have succeeded in growing iPS heart muscle cells in ordered,

Photo: ZEBET / Manfred Liebsch

represent a significant increase in bureaucracy. To deal with this, the Max Planck Institute in Bad Nauheim has taken on extra staff. Braun says that the situation is creating a flood of paperwork, which the regional government offices take a correspondingly long time to process. Meanwhile, deliberations on how to resolve the entrenched conflict with opponents of animal experiments are taking place at the highest level within the Max Planck Society (MPG). This was partly triggered by the incident involving Tübingen-based brain researcher Nikos Logothetis, who halted his experiments on apes in 2015 after receiving threats for months. Another brain researcher, Wolf Singer of Frankfurt, is heading an international commission that has been specially convened by the President of the MPG. “The commission is currently discussing what methods can help overcome the polarization,” says Braun.

Max Planck Institute for Heart and Lung Research MPI-HLR

The Max Planck Institute for Heart and Lung Research, located in Bad Nauheim, investigates developmental processes of organs in the cardiovascular system and the lung.

A second focus is molecular and cellular processes during the formation of diseases in heart, blood vessels and lung, including remodeling processes in these organs. Scientists at the institute search for new approaches to support repair and regeneration of the affected organs.

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The MPI closely cooperates with universities in Frankfurt, Gießen and Marburg. It has become a major part of various federal and state excellence initiates and contributes to two “Gesundheitsforschungszentren”.

Max Planck Institute for Heart and Lung Research W.G. Kerckhoff-Institut Ludwigstr. 43 61231 Bad Nauheim www.mpi-hlr.de



highly organized 3D networks. The team from Hamburg generates heart muscle strips that can be used to test the effect of pharmaceuticals on heart strength.

A team led by Jürgen Hescheler of the Institute of Neurophysiology at the University of Cologne has bathed cardiomyocytes produced by iPS techniques in a toxic substance and then identified molecular biomarkers that are characteristic of cardiotoxicity. The findings come from a project of the EU consortium DETECTIVE, which is supported in part by the European Cosmetics Association (Colipa).

Working towards the ten-organ chip

In vitro models still have their limits when it comes to assessing the efficacy and safety of substances. For example, they are unable to depict the interaction of organs. In an important new development, scientists are now try-

ing to model various connected human organ systems on biochips. Biotechnologists led by Uwe Marx and Roland Lauster of the Technische Universität Berlin and the company TissUse are using iPS technology to engineer human organoids that mimic even the smallest functions of a particular organ. On the chip the human organs are shrunk by a factor of 100,000. The organoids thrive in small chambers on a plate the size of an object slide. The construct is supplied with a fluid-filled system of micro-channels that resembles the bloodstream. The first of these “multi-organ chips” were dual combinations of the skin and liver or the liver and nerve tissue. The most advanced product to come out of the Berlin project is currently a four-organ chip consisting of intestine, liver, kidney and skin modules. The tissue engineers hope to present a multi-organ chip with more then 10 organs in 2018. The research is supported by the BMBF within the GO-Bio-Program. The pharmaceutical industry already regards this miniaturized human test dummy as having great potential for the preclinical research of the future.

Text: Philipp Graf

RetroNectin® can save your cells!

Target cell

RetroNectin Reagent enhances viral transduction by promoting the co-localization of lenti- or retrovirus with target cells. It has been used with great success in sensitive and hard-to-transduce cells such as hematopoietic cells, B cells and T cells. RetroNectin is a recombinant human fibronectin fragment containing three functional domains, two for cell binding and one for binding viral particles.

Cell binding domain

Lentivirus or retrovirus

(C-domain)

RetroNectin

Heparin binding domain (H-domain) CS-1 site (binds to VLA-4 integrin receptor)

Highly efficient in hard-to-infect cells

RetroNectin Supports High-Efficiency Gene Transfer1

• Improves gene transfer efficiency in hard-to-infect cell types and stem cells • Multivalent molecule simultaneously binds virus particles and cell surface proteins, maximizing cell-virus contact • Low toxicity allowing a high cell survival rate

Cell Type

www.clontech.com/retronectin 1 2 3


Viral receptor

VLA-5 (integrin receptor)

Efficiency of Gene Transfer (%)

Human CD34+ CD38– BMC2

95.5

Human PBMC3

91.2

TF-1

97.9

SupT1

97.3

Jurkat

80.1

K-562

90.4

HL-60

86.1

Monkey CD34+ BMC

72.0

Monkey CD4+ T-cell

85.0

Transductions were performed using the RetroNectin-Bound Virus (RBV) Method of transduction. Bone marrow cells. Peripheral blood mononuclear cells.

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ETHICAL ASPECTS

Genome editing in the germline

Controversial cuts In the hands of molecular biologists, genome editing is a powerful tool for the precise engineering of genetic material, something that scientists have long desired. But the revolutionary technology currently taking biomedical laboratories by storm is throwing up ethical and legal questions about the responsible limits of its use. 2015 was the year in which scientific academies worldwide launched an intensive debate on setting guidelines for the use of genome editing. Central to the discussion is the engineering of egg cells, sperm cells and newly created embryos in what are known as germline experiments. German stem cell researchers have stated their position in the debate.

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enome editing is now an established part of basic biomedical research. The designer nuclease system CRISPR/Cas9 took just three years to conquer biomedical laboratories (see GSCN Annual Magazine 14/15). This technology can be used to edit the genetic material in every conceivable type of cell – not just somatic cells but also germline cells, which include egg cells, sperm cells and embryos in the early stage of development. If their genome is edited, this produces a heritable modification that can be passed on to progeny. As exciting as the possibilities of genome editing may seem, the technique is not free from possible errors and risks. Last year saw the unfolding of a scientific debate on the medical applications of genome editing. While the debate was initiated primarily by researchers in the U.S., it was quickly taken up in Germany too.

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Call for a moratorium The debate was triggered by thought-provoking appeals from top researchers in the journals Science and Nature in March 2015. Leading molecular biologists and bioethicists called for a worldwide moratorium on human germline engineering in reproductive medicine. The International Society of Stem Cell Research (ISSCR) and the German Stem Cell Network (GSCN) also spoke out in favor of a moratorium on clinical germline experiments. The German academies of science likewise turned their attention to the issue. In July, the interdisciplinary Gene Technology Report research group at the Berlin-Brandenburg Academy of Sciences (BBAW) produced a highly regarded analysis of human genome engineering that elucidated the ethical and legal aspects of the new technology and interventions in the germline. The paper states that germline therapies and artificial modification of the genome in germline cells are in principle prohibited in Germany under the Embryo Protection Act – but there are a number of loopholes and exceptions. For example, purely in vitro experiments on human germ cells are permitted. There is also no ban on producing germ cells from iPS cells as the law does not cover this relatively new technology. The authors of the BBAW paper support the call for an international moratorium on germline experiments. They hope that during the self-imposed suspension scientists

Max Planck Institute for Molecular Genetics MPIMG

Research at the Max Planck Institute for Molecular Genetics (MPIMG) concentrates on genome analysis of man and other organisms to contribute to a global understanding of many biological processes in the organism, and to elucidate the mechanism behind many human diseases.

It is the overall goal of all MPIMG’s groups to gain new insights into the development of diseases on a molecular level, thus contributing to the development of cause-related new medical treatments. In this context, stem cell research is Annual GSCN Magazine 2015/16

MPIMG

gaining increasing importance. In particular, MPIMG researchers are working on a better understanding of gene regulation networks for tissue formation and homeostasis, as their dysfunction may result in numerous diseases Max Planck Institute for Molecular Genetics Ihnestraße 63-73 14195 Berlin www.molgen.mpg.de

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and academics will engage in open, transparent and critical discussion of the experimental, ethical and legal aspects of germline therapy, elaborate the opportunities and risks of the technology for humans and nature, and draw up recommendations on future regulation. In September, the German National Academy of Sciences Leopoldina and the German Research Foundation (DFG) expressed similar views. Genome engineering summit in Washington

The scientific debate reached an interim climax at the International Summit on Human Gene Editing held in Washington in early December. The conference was organized by the national science academies of the United States, the United Kingdom and China; among the members of

Stem cell researcher Albrecht Müller of the University of Würzburg has closely followed the recent debate on genome editing and has attended many related events, including the summit in Washington. Müller doubts that genome editing will ever be used to correct an unwanted mutation. “That will probably be done by means of pre-implantation diagnostics (PID),” he says. PID involves taking one cell from each of several embryos produced by in vitro fertilization and examining their genetic material. This enables embryos that are free from genetic defects to be iden-

Photo: MHH / Mania Ackermann

Colony forming unit of the myeloide lineage from stem cells

the planning committee were David Baltimore and German science manager Ernst-Ludwig Winnacker. After three days of wide-ranging input and debate, the conference delegates reached the conclusion that basic and clinical research into genome editing of somatic cells must be pursued. With regard to the clinical use of germline editing, the closing statement upholds the existing reservations, stating that because of unresolved safety issues it would be irresponsible to proceed, especially given the lack of a broad societal consensus. However, the dialogue is set to continue, and some national academies have started to draw up guidelines on genome editing.

Leibniz Institute for Zoo & Wildlife Research (IZW) in the Forschungsverbund Berlin e.V. Evolutionary wildlife research for conservation

The Leibniz Institute for Zoo & Wildlife Research (IZW) is an internationally renowned research institute of the Leibniz Association. With the mission of „understanding and improving adaptability“ it examines evolutionary adaptations of wildlife and its resilience to global change, and develops new concepts and measures for conservation. To achieve this, the IZW uses its broad interdisciplinary expertise in evolutionary ecology and genetics, wildlife diseases, reproductive biology and management in a close dialogue 50

with stakeholders and the public. The Department of Reproduction Management, headed by Prof Dr Thomas B Hildebrandt, develops cellular techniques for conservation. This new approach is an essential, promising option for highly endangered wildlife. For this purpose, the IZW builds up international networks on „cellular techniques“, including international expert meetings of the world‘s leading scientists in the field of „cellular techniques“, and develops new methods of securing and preserving wildlife tissue samples.

Leibniz-Institut für Zoo- und Wildtierforschung (IZW) im Forschungsverbund Berlin e.V. Alfred-Kowalke-Straße 17 · 10315 Berlin

www.leibniz-izw.de


ETHICAL ASPECTS

tified. “Genome editing for monogenic disorders makes little sense, and for polygenic ones it makes no sense at all,” comments Müller. He reports that a possible application that was discussed in Washington is human enhancement, which involves modifying traits such as skin color. According to Müller, the conference participants were clear that such techniques are ethically highly controversial. Further discussion needed

With regard to germline experiments in Germany, Müller believes that there must be a very clear boundary: no genetically modified embryo should be implanted in a womb. (Implantation is in any case banned under the Embryo Protection Act.) “But researchers should be able to use embryos left over from PID in their genome editing work,” says Müller. He is of the view that the present discussion must be pursued by scientists and society at large. He maintains that “the solutions must come from science itself; the law shouldn’t act prematurely, as happened with the Stem Cell Act. We are just at the beginning of the road here.”

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Hans Schöler, Director of the Max Planck Institute for Molecular Biomedicine in Münster, is highly critical of genome editing in the germline. He currently rejects intervention in human embryos, even for research purposes. “It doesn’t seem to me at the moment that off-target effects can be controlled sufficiently for errors to be excluded,” says Schöler. He points out that his own experiments on mouse cells have shown that even with recent versions of the CRISPR/Cas system a large number of faulty cuts in the genome still occur. In a germline experiment, an embryo is doubly stressed since its genes must be both checked and corrected. “In my view it would make more sense to test several embryos and transfer one that has no genetic defects.” However, if the technique of genome editing were perfected and there was a valid reason for using it – instead of PID – Schöler says he might be able to accept it. However, he points out that this is nowhere near being the case at present. Nevertheless, he believes that now is the right time for public discussion of the ethically controversial matter of intervention in the human genome. Text: Philipp Graf

Im Zentrum der AMSBIO Kernprinzipien stehen Qualität und Innovation, weshalb wir uns ständig um neue, innovative Produkte bemühen, die helfen Ihr Stammzellprojekt voranzutreiben. Zur technischen Unterstützung unserer Produkte steht Ihnen unser Expertenteam zur Verfügung. Wir sind in der Lage unsere Fachkompetenz zur Unterstützung Ihrer Forschungsprogramme direkt durch unsere Auftragsforschungslabors umzusetzen und können dadurch spezifisch auf Produktanforderungen und Kundenwünsche eingehen.

Besuchen Sie www.amsbio.com oder schreiben Sie an 51

Photo: Photolia / tarasov-vl

ESSAY | UNGEPRÜFTE STAMMZELLTHERAPIEN

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ESSAY: UNTESTED STEM CELL THERAPIES

ESSAY: Untested stem cell therapies

Quacks and charlatans: Age-old adversaries, but no less dangerous!

by Ira Herrmann The process for developing new drugs and treatment methods is usually lengthy and full of obstacles. In the interests of effectiveness and safety, research and validation are conducted in a way that allows them to be as internationally comparable as possible. By now, guidelines and standards on the manufacturing and testing of drugs and pharmaceutical products are subject to international regulations in order to ensure patient safety and the efficacy of new medicines. Nevertheless, some forms of treatment that are well established and fully approved today began life in “compassionate use” – i.e. outside controlled clinical studies. And there have always been those who have tried to bypass the lengthy clinical development stage so as to be able to offer new therapies faster. This secures them, at best, a shady chapter in medical history, and usually the only tangible results are their fatter wallets.

Regardless of the drug or procedure that needs to be tested, one of the constant challenges that developers and practitioners face is how to deal with patient expectations. This is a particularly heavy burden when it comes to treating patients with very limited options and, in the worst cases, low chances of survival. For some years now, certain providers have been taking advantage of the hype surrounding stem cells as some kind of miracle cure and exploiting the fact that there are currently no stem cell-based treatments and products available and that the legal framework for the use of stem cells is often sketchy. Yet the history of medicine has always been full of “miracle healers” and charlatans. What’s more, loopholes in medical law are often knowingly accepted in the interests of freedom of therapy and the development of new fields. So why is there such a fuss about offering unproven stem cell therapies? And is the reaction justified?


• Due to their special properties, stem cells have the potential to treat a wide variety of illnesses – including many widespread and/or currently incurable diseases. This potential creates expectations that science is unable to meet in such a short period of time (just as a reminder: the first isolation of human embryonic stem cells was achieved in 1998).

• Stem cell research is a young discipline with its roots in developmental and cell biology. Today, however, it needs knowledge input from other natural scientists, engineers and, above all, representatives of various medical and clinical disciplines. It has thus become a truly interdisciplinary scientific field. However, this interdisciplinarity also requires the establishment of a common language when discussing basics, methods and aims.

• The approval of stem cell-based therapies is subject to new and very particular legal conditions. So far, only very few authorizations have been granted worldwide, giving us very few precedents to learn from. In Europe, for instance, only six products have been approved under the EU Regulation on advanced therapy medicinal products (ATMPs), which came into force in 2008. So there is also still (pioneering) work to be carried out in this area by developers and regulatory authorities.

So we see that stem cell research is a young biomedical discipline that has great potential and capacity for innovation, but that is confronted with extreme expectations. These expectations need to be addressed and worked through in line with the principles of good scientific practice. It is advisable, therefore, to conduct dialogue with all stakeholders – dialogue that also allows for the public criticism of providers of unproven stem cellbased treatments. Not because quacks and charlatans are a new phenomenon, of course, but because stem cell research is such an important new field with the potential to solve the problems of an ageing society. 53

GSCN ANNUAL REPORT

GSCN

Annual Report Boards

Executive Board

In accordance with Section 8(1) of the statute and rules of the German Stem Cell Network (GSCN) e.V., the Executive Board (presidium) consists of the Acting President (chairperson), the Senior President (first deputy chairperson), the Designated President (second deputy chairperson), the Treasurer, and the Assessor. In the reporting period (Sept. 2015 to Sept. 2016), the Executive Board is made up of the following members: Acting President (Chair)

Prof. Ulrich Martin (Hannover Medical School) E-mail: [email protected]

Designated President (2nd Vice Chair)

Prof. Karl Lenhard Rudolph (Leibniz Institute on Aging – Fritz Lipmann Institute [FLI], Jena) E-mail: [email protected]

Senior President (1st Vice Chair)

Treasurer

Assessor

Prof. Thomas Braun (Max Planck Institute for Heart and Lung Research, Bad Nauheim) E-mail: [email protected]

Dr. Michael Cross (Leipzig University) E-mail: [email protected]

Prof. Frank Emmrich (Fraunhofer IZI and Leipzig University) E-mail: [email protected]

The Executive Board in 2015

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The GSCN would like to thank the members of the 2015 Executive Board (Nov. 2014 to Sept. 2015). Andreas Trumpp, Active President in 2014, left the Executive Board as scheduled and has now been elected as a member of the Extended Board.

Photo: GSCN

Executive Board members: (from left): Karl Lenhard Rudolph, Thomas Braun, Ulrich Martin, Frank Emmrich, Michael Cross, Andreas Trumpp


GSCN ANNUAL REPORT

GSCN Executive and Extended Boards and Management

Extended Board

Photo: GSCN

In accordance with Section 9(1) of the statute and rules of the German Stem Cell Network (GSCN) e.V., the Extended Board consists of up to 15 members. In the reporting period, the Extended Board is made up of the following members: Prof. Oliver Brüstle (University of Bonn) E-mail: [email protected]

Dr. Tobias Cantz PD (Hannover Medical School) E-mail: [email protected]

Dr. Micha Drukker (Helmholtz Zentrum München) E-mail: [email protected]

Prof. Hartmut Geiger (Ulm University) E-mail: [email protected]

Prof. Magdalena Götz (LMU Munich) E-mail: [email protected]

Ira Herrmann (Stem Cell Network North Rhine Westphalia; Life & Brain since 1 Jan. 2016) E-mail: [email protected]

Prof. Ana Martin-Villalba (German Cancer Research Center Heidelberg) E-mail: [email protected]

Prof. Albrecht Müller (University of Würzburg) E-mail: [email protected]

Prof. Michael Rieger (Goethe University Frankfurt) E-mail: [email protected]

Prof. Ingo Roeder (TU Dresden) E-mail: [email protected]

Prof. Hans R. Schöler (Max Planck Institute for Molecular Biomedicine) E-mail: [email protected]

Prof. Wolfgang Wagner (RWTH Aachen) E-mail: [email protected]

Prof. Andreas Trumpp (German Cancer Research Center Heidelberg) E-mail: [email protected]

Prof. Mathias Treier (Max Delbrück Center for Molecular Medicine Berlin) E-mail: [email protected]

Prof. Claudia Waskow (TU Dresden) E-mail: [email protected]


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GSCN ANNUAL REPORT

Working group initiators Scientific working groups Pluripotency and Reprograming

Somatic Stem Cells and Development

Dr. Micha Drukker Prof. Hans Schöler Prof. Mathias Treier

Prof. Thomas Braun Prof. Elly Tanaka

Basic, Translational Prof. Claudia Waskow and Applied Hematopoiesis Prof. Timm Schroeder (ETH Zürich) E-mail: [email protected]

Stem Cells in Diseases Prof. Thomas Brabletz (University Medical Center Freiburg) (Cancer Stem Cells) E-mail: [email protected] Prof. Andreas Trumpp Stem Cells in Regenerative Therapies

Stem Cells in Disease Modeling and Drug Development

Dr. Michael Cross Prof. Ulrich Martin

Prof. Oliver Brüstle Prof. Karl-Ludwig Laugwitz (University Hospital Klinikum rechts der Isar, Munich) E-mail: [email protected]

Computational Stem Cell Biology

Prof. Georg Füllen (Institute for Biostatistics and Informatics in Medicine and Ageing Research, Rostock) E-mail: [email protected] Prof. Ingo Röder (TU Dresden) E-mail: [email protected]

Strategic working groups

Funding Programs and Policies

Prof. Ulrich Martin Prof. Albrecht Müller Acting GSCN President (ex-officio)

Public Engagement and Outreach Activities

Dr. Tobias Cantz PD Ira Herrmann

Promotion of Young Researchers Prof. Hartmut Geiger (Ulm University) E-mail: [email protected] Prof. Jürgen Hescheler (University Hospital Cologne) E-mail: [email protected] Dr. Insa Schröder (GSI Helmholtz Centre for Heavy Ion Research) E-mail: [email protected] Patient Information Dr. Gisela Badura-Lotter (Ulm University) (Stem Cell Therapies) E-mail: [email protected] Ira Herrmann

Clinical Trials and Dr. Zoltán Ivics (Paul-Ehrlich-Institut, Langen) Regulatory Affairs E-mail: [email protected] Dr. Andreas Kurtz (BCRT Berlin), E-mail: [email protected] Prof. Torsten Tonn (Institute for Transfusion Medicine, Dresden) E-mail: [email protected] Prof. Hans-Dieter Volk (BCRT Berlin) E-mail: [email protected]

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Stem Cell Technologies Dr. Andreas Bosio (Miltenyi GmbH, Bergisch Gladbach) E-mail: [email protected] Prof. Frank Emmrich


GSCN ANNUAL REPORT

Facts and figures Meetings

Executive Board meetings

The Executive Board of the GSCN regularly holds meetings and telephone conferences. These meetings are coordinated and organized by the Central Office. The following Executive Board meetings took place in 2015: • Video conference (19 May 2015) • Video conference (15 July 2015) • Meeting (9 Sept. 2015) in Frankfurt am Main

Extended Board meeting

• 9 Sept. 2015 in Frankfurt am Main

General Assembly

• 10 Sept. 2015 in Frankfurt am Main

Overview of members in 2015 Total no. of members Natural persons

Legal persons

334 Full members

189

Junior members

116

Research institutes

17

Companies with more than 20 full-time staff

4

Companies with fewer than 20 full-time staff

7

Partner societies

1

Membership cancelations in 2015

32

Members of the working groups Scientific working groups

Pluripotency and reprograming

184

Somatic stem cells and development

145

Basic, translational and applied hematopoiesis Stem cells in diseases (cancer stem cells)

136

Stem cells in regenerative therapies

137

Stem cells in disease modeling and drug development

185

Computational stem cell biology Strategic working groups

64

24

Funding programs and policies

124

Career development

111

Clinical trials and regulatory affairs

96

Public engagement and outreach activities

60

Patient information (stem cell therapies)

17

Stem cell technologies

178

Last updated 31.1.2016


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GSCN ANNUAL REPORT

Institute members

Company members

• Berlin-Brandenburg Center for Regenerative Therapies (BCRT)

• AMS Biotechnology (Europe) Ltd. • Biological Industries • Eppendorf AG • Essen BioScience Ltd. • Hexcell Berlin GmbH • HI-STEM gGmbH • Thermo Fisher Scientific • Macopharma Int. GmbH (from 2016) • PELOBiotech GmbH • PeproTech GmbH • Takara Bio Europe S.A.S.

• Center for Regenerative Therapies Dresden (CRTD)

• German Cancer Research Center (DKFZ), Heidelberg

• Fraunhofer Research Institution for Marine Biotechnology (Fraunhofer EMB), Lübeck

• Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Aachen • Fraunhofer Institute for Production Technology (IPT), Aachen (from 2016)

• Fraunhofer Institute for Cell Therapy and Immunology (Fraunhofer IZI), Leipzig • Institute of Reconstructive Neurobiology, University of Bonn

• Institute for Stem Cell Research and Regenerative Medicine, Düsseldorf • Leibniz Institute on Aging – Fritz Lipmann Institute (FLI), Jena • Leibniz Institute for Zoo and Wildlife Research in the Forschungsverbund Berlin

• Max Delbrück Center for Molecular Medicine (MDC), Berlin-Buch • Max Planck Institute for Heart and Lung Research (MPI-HLR), Bad Nauheim • Max Planck Institute for Molecular Genetics (MPIMG), Berlin • REBIRTH Cluster of Excellence, Hannover Medical School

• Collaborative Research Center SFB 873, Centre for Organismal Studies, Heidelberg University Hospital

Partner societies • German Society for Transfusion Medicine and Immune Hematology (DGTI) e.V..

• Tissue Engineering and Regenerative Medicine (TERM), University Hospital of Würzburg

• Universitätsklinikum Erlangen, Department of Molecular Neurology, Bavarian Research Network ForIPS – Induced Pluripotent Stem Cells (from 2016)

GSCN General Assembly

General Assembly 2015 58

Photos: GSCN

Eighty-eight members attended the third General Assembly of the GSCN, held on 10 Sept. 2015 during the 3rd Annual Conference in Frankfurt am Main. The minutes of the assembly and the presentation that formed part of it can be downloaded from the members’ area of the website. The Executive Board and the Extended Board were re-elected in the online elections of August 2015 (see above) after the auditors officially discharged the previous board members.


GSCN ANNUAL REPORT

Activities in 2015

2015 was a busy year for the GSCN, with numerous events, new formats, and interesting workshops and conferences. On the one hand, the GSCN is a members’ association with a focus on networking; on the other, it promotes communication with the public and policymakers. Below is a summary of the activities undertaken by the GSCN in both of these areas.

The network

The GSCN Annual Conference is the largest assembly of stem cell researchers. At the 3rd International Annual Conference of the GSCN in Frankfurt am Main, 400 scientists came together to exchange ideas, develop partnerships, initiate new projects, and bring each other up to date on the latest research. Networking is about making contacts, developing collaborative projects, establishing friendly relationships, and linking new research to new faces and institutions. These networks are of ongoing importance to current and future projects. The GSCN has also created contact platforms on the international level. For example, it developed a new format called the WunderBar Evening for the ISSCR Annual Meeting in Stockholm on 23 – 26 June 2015. Around 100 members and friends attended the event held on the roof terrace of the Clarion Hotel Sign at the close of the conference day. The evening proved to be a success despite the cool Scandinavian temperatures, with great midsummer views over Stockholm, a fun and relaxed atmosphere, delicious finger food and drinks, and stimulating conversation. The GSCN Meet-Up Hub took place the following afternoon in the ISSCR exhibition hall. As well as GSCN members, the event also attracted many international colleagues for an inspiring one-hour meet-up in the middle of the conference. “I’m very satisfied with both events; we achieved our goal of creating spaces for people to network in a pleasant and animating environment,” said Daniel Besser, Managing Director of the GSCN. Twenty-one junior scientists who had been granted Travel Awards by the GSCN working groups attended the meet-up – and their response was particularly enthusiastic. “For me, it was great to attend the GSCN Meet-Up Hub. I met several scientists who were working in Germany. We discussed future directions of the field of pluripotency and reprograming as well as potential collaborative works,” said award winner Kee-Pyo Kim of the Max Planck Institute for Molecular Biomedicine in Münster. Scientists at all stages of their careers are involved in the GSCN network. This year, the GSCN organized the first round of the GSCN Awards to raise awareness of the work carried out by specific groups in these various stages and to recognize outstanding achievements. For each of the awards, there is a submission process that includes stating the reasons for nomination, convening a selection committee, and


Participants of the Non-PI-Meeting in Bonn

plenty of work on the part of the jury. The following scientists received awards for their outstanding and impressive research work: - The GSCN Young Investigator Award was granted to Dr. Julia Ladewig of the Institute of Reconstructive Neurobiology at University Hospital Bonn. - The GSCN Female Scientist Award was presented to Prof. Magdalena Götz of the Institute of Stem Cell Research at Helmholtz Zentrum München. Götz is also chairs the Department of Physiological Genomics at LMU Munich.

- The GSCN Publication of the Year Award went to Jichang Wang and Dr. Zsuzsanna Izsvák from the Mobile DNA research group at the Max Delbrück Center for Molecular Medicine (MDC) in Berlin-Buch for their article “Primate-specific endogenous retrovirus-driven transcription defines naïve-like stem cells”, which was published in the academic journal Nature (Wang, J. et al., 2014, Nature, doi:10.1038/nature13804). The GSCN Awards ceremony took place at the Presidential Symposium held at the GSCN Conference in Frankfurt and featured talks by the three award-winners, who all felt deeply honored to have received the awards (see Conference report). For the GSCN, the awards ceremony represents a commitment to promoting science policy. In 2015, the GSCN again granted Travel Awards to enable talented junior scientists to take part in various conferences and summer/winter schools. Thanks to the Travel Awards, Onur Basak (Hubrecht

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GSCN ANNUAL REPORT

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Communication Cultivating contacts, collaborating, and communicating in a network are important ways of passing on information. The GSCN makes use of various communication channels. At the conference the annually updated mobile app GSCN Navigator enables participants to shape their visit to the conference to suit their personal interests. The GSCN also provides information and news in German and English on its website www.gscn.org. In 2015 the website attracted more than 24,700 visits from 15,000 users (2014: 18,000 visits from 9,500 users). Visits have thus increased by 28% and the number of users by 37%. The news column on the website’s home page highlights the latest developments in the field – a service that is appreciated by many users. The GSCN Newsletter goes out by e-mail and the Annual Magazine, which focuses on different topics each year, is sent out to members in the mail. In 2015 the magazine concentrated on stem cell technologies, considering Germany from the perspective of the various core institutions involved. The 2015 annual magazine was very well received and a record 1,000 copies were distributed. At the first GSCN annual conference in Berlin in 2013, members expressed a wish for an internal, web-based communication platform: in 2015 this was created in the form of the GSCN Humhub. This social Intranet has not yet become established as a fully-fledged professional and social stem cell network – but of course this is the goal. Each of the scientific and strategic working groups has its own forum; members are assigned to their own working groups but can also participate in other groups. This is intended to provide a very simple, forum-like means of communication that enables members to discuss issues online, exchange information, to post details of conferences, workshops, talks and publications, and to debate these. As a bonus, promotional codes for conferences, grants or other member benefits will appear first on Humhub. 2015 was the launch year – the GSCN Humhub network should really get underway in 2016. Internationally, the GSCN worked closely with EuroStemCell in 2015. At a wide-ranging European workshop in Brussels (18 – 19 Feb. 2015), the EuroStemCell team set out new objectives, and topic groups were formed to drive forward this European information portal and promote the working groups behind the new funding application to the European Commission. In 2015 most of the work

Photos: GSCN

Institute, Utrecht, Netherlands), Debojyoti Chakraborty (BIOTEC, University Hospital Carl Gustav Carus, Dresden), Christina Galonska (Broad Institute of MIT and Harvard, Cambridge, USA), Sarah Konze (Hannover Medical School), and Miha Modic (Helmholtz Zentrum München) were able to attend the GSCN Annual Conference in Frankfurt. All Travel Awards were funded by GSCN company member Eppendorf AG. The GSCN continued to promote young talent and provide financial, subject-related and logistical support through the first non-PI meeting in Bonn (20 April 2015), where 18 young scientists came together for an exchange of ideas in a series of lectures and discussions. “All the participants were enthusiastic about the atmosphere of the meeting and the possibility of sharing their experiences with one another. They encourage the GSCN to continue the concept of non-PI meetings in the future,” said participants Adele Marthaler and Henning Kempf. The non-PI meeting took place in the run-up to the 8th International Meeting of Stem Cell Network North Rhine-Westphalia in Bonn (21 – 22 April 2015). After the meeting, the GSCN was the main sponsor of the 6th Bioinformatics and Stem Cells Satellite Workshop (22 – 23 April 2015), a one-day event organized by Georg Füllen, initiator of the Computational Stem cell Biology working group. The strategic working group on Career Development explored a completely new field by organizing a workshop on “Scientists in Management” in Darmstadt (17 – 18 Nov. 2015). It is especially useful for scientists to master soft skills like strategic and management competencies at an early stage in their careers. “The response to the workshop was extremely positive,” said Insa Schröder, initiator of the working group on Career Development. In addition, the GSCN presented information materials at the World Conference on Regenerative Medicine in Leipzig (21 – 23 Oct. 2015; see News) and various other meetings and conferences, such as the IGLD Meeting at the Max Delbrück Communications Center in Berlin (12 – 14 March 2015) and the 4th International Conference on Strategies in Tissue E ngineering (SITE) in Würzburg (10 – 12 June 2015). The GSCN played a key role in establishing a network of German stem cell core units in a bid to stimulate discussion and improve communication between scientists. The goal of the new network is, first and foremost, to create and standardize joint quality protocols and methods to safeguard the pluripotent state of hiPSCs.


GSCN ANNUAL REPORT

Poster exhibition in the Urania, Berlin

took place behind the scenes, but in 2016 and 2017 this major European partner will launch many new projects in a new guise – this will include a relaunch of the website, which the GSCN is helping to design. The relationship between the GSCN and EuroStemCell is fruitful for both organizations – for example, the GSCN used the Edinburgh meeting factsheets on various diseases (www.eurostemcell.org) to produce a colorful poster display that is proving popular at events.

Photos: GSCN

These communicators’ meetings also promote networking on international stem cell research. The international collaboration was continued at the meeting in Edinburgh on 12 – 13 Nov. 2015. Its subject was “Train the Trainers,” and it provided many examples of how information on stem cell research can be communicated in a way that inspires interest in people of all ages and educational backgrounds. Nationally, the GSCN organized a similar meeting with its Communication on Stem Cell Research group. Held in Berlin on 27 – 28 April 2015, the meeting combined a workshop on public affairs with networking and the development of new projects. Like last year’s meeting in Hannover (which we reported on), the event generated many good ideas and helped to develop useful contacts between the science communicators of the various research institutions. One specific outcome was the defining of the group of institutes that will organize UniStem Day on 11 March 2016 as a European day of action on stem cell research. A thousand school students are expected to take part in a day of concentrated activities covering all aspects of stem cell research – including talks, films, games and guided tours.

School students were the target audience of GSCN events at the MINT400 forum at the Max Delbrück Center (MDC) in Berlin (12 - 13 Feb. 2015 and 4 - 5 Feb 2016), at the Science Day at the Robert-Havemann-Gymnasium in Berlin-Karow (19 Nov. 2015) and at the debate on stem cells and ethics at Haus Kreisau youth education center (29 Jan. 2015). The MDC in Berlin puts on regular informational careers


events for young scientists wanting to learn more about new professional fields. At the knowledge management careers fair on 20 Nov. 2015 Daniel Besser explained the duties of the managing director of a scientific network and discussed his work with the young scientists who attended.

The GSCN organized games and talks for interested members of the public at the Long Night of the Sciences in Berlin on 13 June 2015. Following on from the Frankfurt conference, the GSCN held a panel discussion on the Westend campus at which facilitator Stefanie Seltmann (DKFZ) and panel members Thomas Braun (MPI for Heart and Lung Research), Hubert Serve (oncologist, hematologist, University Hospital Frankfurt), and Andreas Zeiher (cardiologist, University Hospital Frankfurt) debated the latest laboratory and hospital research, taking as their subject the use of stem cells in the treatment of heart attacks and leukemia. The latest GSCN films about Andreas Zeiher and Hubert Serve were also shown. Like the GSCN films about Magdalena Götz, Andreas Trumpp and Anthony Ho from 2014, these have now been

MINT-Students spend a „stem cell day“ with Daniel Besser 61

GSCN ANNUAL REPORT

GSCN panel discussion in Frankfurt: Thomas Braun, Hubertus Serve, Stefanie Seltmann and Andreas Zeiher debate about possible applications of stem cell therapies.

The year’s events drew to a close on 2 Dec. 2015 with the GSCN panel discussion on “Modern cell therapies – Using stem cells to treat heart and blood disorders” at the Urania in Berlin, where the audience enjoyed screenings and a panel discussion with Claudia Waskow (hematologist, TU Dresden), Daniel Besser (Managing Director, GSCN), and Carsten Tschöpe (cardiologist, Charité and Berlin-Brandenburg School for Regenerative Therapies). At the World Health Summit in Berlin on 11 – 13 Oct. 2015 the GSCN again put the subject of stem cells on the agenda by organizing its own session. A panel of high-profile international speakers discussed the future applications and potentials of stem cells, thus bringing the research field to the attention of an audience of medical experts and policymakers from all over the world.

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The GSCN is also active in social media with a growing number of followers. The GSCN Central Office regularly receives enquiries from teachers, students, patients and journalists, which are answered with the help of scientists. The GSCN conference, in particular, prompts nationwide coverage in print and online media.

Finances

The GSCN is a non-profit organization funded by membership subscriptions and grants from the BMBF. Under Section 4 of its statute, the level of subscriptions is set by rules adopted at the General Assembly. Subscriptions are detailed on the GSCN’s membership form. The business year is the calendar year. Subscriptions are due at the start of the business year. The Executive Board is responsible for producing the annual accounts and submitting them to the General Assembly. Details of the association’s finances are provided at the General Assemblies.

Photos: GSCN

translated into English and are proving to be very popular items on the EuroStemCell website. They have the highest click rates on the European website. The films can be found on the GSCN website at: www.gscn.org/en/RESOURCES/GSCNMovies.aspx.


Photo: GSCN



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