IMMUUN Volume 4 - Issue 1 - March 2015 -
for every professional in the immunology chain
Ton Schumacher: â€œImmunotherapy has become a valuable treatment option for cancerâ€? How to catch Circulating Tumour Cells Immunity and Science Fiction: The next fifty years of immunology research
Theme Immunology and
Breaking new ground
Lia van der Hoek Towards identification of all human viruses
Community of open innovation What we are
More than just a workplace! Pivot Park is the pivot in a flourishing network of companies and knowledge institutions involved, either directly or indirectly, in innovative research in the field of life sciences. We provide accommodation to a diverse range of companies with specialised knowledge in an open innovation culture. Pivot Park provides ambitious companies in the life sciences with access to the entire spectrum of pharmaceutical R&D.
This open innovation and partnership extends beyond the gates. Partners include companies and the best departments of universities in the vicinity. Pivot Park is not affiliated with any one university or knowledge institute. We aspire to forge partnerships on the broadest possible scale, from the local to the international level: an open community without borders!
What we offer An exciting environment with a high quotient of collaboration and diversity. New and existing life science companies have every opportunity here to be inspired by one another through networking sessions, project-based partnerships or scientific conferences. The end result is a stimulating cross-pollination that produces valuable input for a healthy society. Pivot Park provides the foundation: an optimal infrastructure, state-of-the-art facilities and a wealth of knowledge and experience.
High quality facilities We have state-of-the-art core facilities that we can open to ambitious companies in the life sciences, thanks to flexibility and customisation, with our high-grade infrastructure providing a strong launching point. This is targeted not only at start-up companies, but also those wishing to take the next step, with their often more complex needs.
contents IMMUUN March2015
Immuun is published by the NVVI and written and edited by Bureau Lorient Communicatie BV. In 2015, Immuun will have three issues. The target groups are NVVI members, relevant clinicians, suppliers and other partners of immunologists as well as policy makers. EDITORIAL BOARD Dr. Godelieve de Bree Prof.dr. Mieke Boots Dr. Hans Jacobs Dr. Edward Knol Prof.dr. Yvette van Kooyk (chair) Dr. Ingeborg Streng-Ouwehand Dr. Andrea Woltman
4 7 18 24 20 28 30 34 News
Lia van der Hoek
Science across borders Schizophrenia
Portrait Passion for research Hidde Ploegh
recommendation committee Prof.dr. R.E. Bontrop Prof.dr. F. Claas Prof.dr. M.R. Daha Prof.dr. C.G.M. Kallenberg Prof.dr. G. Kraal Prof.dr. R.A.W. van Lier Prof.dr. C.J.M. Melief Prof.dr. D. Roos Prof.dr. J. van de Winkel editor in chief Drs. L. van der Ent PUBLISHER NVVI Contacts via Bureau Lorient Communicatie BV Hoofdstraat 98 - 100 2235 CK Valkenburg ZH T + 31 71 5890848 email@example.com lay out & print Van der Weij BV Grafische Bedrijven, Hilversum
Cancer Vaccine Tracking project Translating immunology to the clinic
NVVI 50th anniversary Immunity and Science Fiction
Compare Fighting infectious diseases
Theme Immunology and cancer
Research Cancer immunotherapy as clinical reality
Jolanda de Vries en Nicoline Hoogerbrugge
PHoto front page Bureau Lorient Communicatie, Lagro Fotografie
The worldâ€™s first preventive cancer vaccine
advertizing Congress Company Bruistensingel 250 5232 AD â€™s-Hertogenbosch T +31 73 7003500 firstname.lastname@example.org DISCLAIMER Immuun is made with the utmost care. NVVI nor Bureau Lorient Communicatie BV can be held responsible or liable for errors. Articles do not necessarily reflect the opinion of the editorial board, the publisher or the writer.
Immuno Valley Award winner
Leon Verstappen Circulating Tumour Cells Detection and removal
Astrid Visser Antigen-specific immunomonitoring
Cancer Immunology research at Pivot Park
dutch society for immunology
Hunt for information Cancer and immunology is undeniably a hot topic –
and rightly so. The concept of helping a person’s own immune system to battle cancer is extremely appealing. Wouldn’t it be great if we could manipulate the human immune system in a way to fight tumours? Research results are promising, fueling hopes for a breakthrough. Patients with a suppressed immune system – e.g. after transplantation or immune deficiencies– are more likely to get cancer than persons with optimally functioning immune response. This is totally congruent with the concept of empowering the immune system to battle cancer. But it must be done in a very targeted way: there is the risk of creating an imbalance. If the immune system would become overactive, we can create auto immune diseases. It is therefore essential to thoroughly understand the functioning of the immune system. In this respect, the research on cancer immunology is extremely valuable as well. Apart from its primary goal, which is to eliminate cancer, it can improve our basic knowledge of the immune system, thus providing us with new tools to manipulate the immune system. The NVVI supports the wish to improve our knowledge of the basic functioning of the immune system.
Two antibodies that are connected have a much more powerful immune reaction against all sorts of unwanted elements in our body than a single antibody. But to make a strong connection is quite difficult. Researchers of the AMC spin-off AIMM Therapeutics have succeeded in connecting two antibodies against the flu, while retaining their function. “The possibilities this method offers are unprecedented”, says Koen Wagner of AIMM Therapeutics. “The benefits of bi-specific antibodies – as two linked antibodies are called – are legion.” You only need to administer something once and it has a much broader function. But to connect the two antibodies is hard. Up to now, they had to be adjusted in such a way that they lost their stability. As a consequence, some antibodies were destroyed within two hours after connection. Wagner learned from researchers in Boston how two antibodies could be fixed with only minimal adjustments. Back in Amsterdam he and his colleague Mark Kwakkenbos applied the method to ‘two top-specimens’ that together can eradicate all type-A flu viruses. Research in mice showed that the combination remains stable and offers good protection, as could be read in a recent publication in the scientific top-magazine PNAS. “Bi-specific antibodies can be used for a lot more goals”, says Kwakkenbos. “For instance in oncology, for fighting tumours. Once you can connect antibodies, they become far more effective.” Wagner: “That goes in fact for everything related to the immune system.” In order to let patients benefit from this development, a lot more work has to be done. AIMM now starts investigating whether the coupled antibodies can be produced on an industrial scale.
It is vital that we exchange information and learn from each other. We can and should do better. Congresses usually focus on specific diseases, while combining the knowledge on the underlying mechanisms that are
Source: AMC Magazine December 2014, PNAS publication, December 11, 2014 - A bispecific antibody generated with sortase and click chemistry has broad antiinfluenza virus activity
operational in various diseases, is equally important. Researchers are inclined to stay inside their disease topics. The NVVI urges researchers to broaden their perspective and to hunt for information in other research areas than their own. It is wise to gain insight into other disciplines. Talk to your colleagues in and outside your own institute and visit a congress in an adjacent field. Cross borders and break down walls. It also works the other way around. Maybe you know an oncologist, a rheumatologist, a haematologist or another specialist who could benefit from a better knowledge on immunology? Encourage him or her to learn more and to actively seek cooperation with you as a specialist in immunology. The NVVI welcomes such requests as well. Reina Mebius, Dutch Society for Immunology
Hans van Eenennaam, BioNovion: “Start thinking from the end back” In the Netherlands we have a huge amount of immunology knowledge. This knowledge gives us an excellent position to make a difference in this field. It would, however, be beneficial to both healthcare and economy when more knowledge would be converted into bedside products. It is common knowledge that there are hiccups in the value chain, but how to identify, address and solve these? BioNovion COO Hans van Eenennaam has a piece of the puzzle: “In many cases, researchers develop knowledge, a target or a technology and only then start thinking of a product. Why not do that the other way round, to start by thinking of an end product all the way back to your own work. Ask clinicians and pharmaceutical companies: what do patients need? What could my contribution mean? What is already available and under development? What do pharmaceutical companies demand from promising discoveries?”
Binge drinking disrupts immune system in young adults Binge drinking in young, healthy adults significantly disrupts the immune system, according to a study led by dr. Majid Afshar of Loyola University Chicago Stritch School of Medicine in Maywood, Illinois. Depending on their weight, study participants drank four or five shots of vodka. Twenty minutes after reaching peak intoxication, their immune systems revved up. But when measured again, at two hours and five hours after peak intoxication, their immune systems had become less active than when sober. The study by pulmonologist, critical care physician and epidemiologist Majid Afshar, MD, MSCR, at the Loyola University Chicago Stritch School of Medicine and colleagues is published online ahead of print in Alcohol, an international, peer-reviewed journal. Many health risks of binge drinking are sufficiently known. “But there is less awareness of alcohol’s harmful effects in other areas, such as the immune system,” said Elizabeth Kovacs, PhD, a co-author of the study and director of Loyola’s Alcohol Research Program. The National Institute on Alcohol Abuse and Alcoholism defines ‘binge drinking’ as drinking enough to reach or exceed a blood alcohol content of .08, the legal limit for driving. This typically occurs after four drinks for women or five drinks for men, consumed in two hours. One in six U.S. adults binge drinks about four times a month, and binge drinking is more common in young adults aged 18 to 34, according to the Centers for Disease Control and Prevention.
Less active Two hours and five hours after peak intoxication, researchers found the opposite effect: fewer circulating monocytes and natural killer cells and higher levels of different types of cytokines that signal the immune system to become less active. Dr. Afshar is planning a follow-up study of burn unit patients. He will compare patients who had alcohol in their system when they arrived with patients who were alcohol-free. He will measure immune system markers from each group, and compare their outcomes, including lung injury, organ failure and death. Majid Afshar, “Acute immunomodulatory effects of binge alcohol ingestion”, in Alcohol, with co-authors Elizabeth Kovacs, PhD, of Loyola and Stephanie Richards; Dean Mann, MD; Alan Cross, MD; Gordon Smith, MPH; Giora Netzer, MD; and Jeffrey Hasday, MD, all of the University of Maryland.
Ramping up Dr. Afshar led the study while at the University of Maryland, where he completed a fellowship before joining Loyola. The study included eight women and seven men with a median age of 27. Each volunteer drank enough shots of vodka – generally four or five – to meet the definition of binge drinking. Dr. Afshar and colleagues took blood samples at twenty minutes, two hours and five hours after peak intoxication because these are times when intoxicated patients typically arrive at trauma centres for treatment of alcohol-related injuries. The blood samples showed that twenty minutes after peak intoxication, there was increased immune system activity. There were higher levels of three types of white blood cells that are key components of the immune system: leukocytes, monocytes and natural killer cells. There also were increased levels of proteins called cytokines that signal the immune system to ramp up.
Stonehenge trouble on stones. Licensed under CC BY-SA 2.0 via Wikimedia Commons - http://commons.wikimedia.org/wiki/File:Stonehenge_trouble_ on_stones.jpg#mediaviewer/File:Stonehenge_trouble_on_stones.jpg
Carsten Linnemann wins Antoni van Leeuwenhoek prize 2015 Carsten Linnemann received this year’s Antoni van Leeuwenhoek prize on Monday, January 12. The prize is awarded annually to an outstanding young researcher at the Netherlands Cancer Institute, in order to promote his or her career. Linnemann’s work focuses on understanding the immune system and stimulating it to destroy cancer cells. This type of cancer therapy, called immunotherapy, has recently known important breakthroughs. Linnemann, who is originally from Germany, obtained his PhD cum laude in 2013 in the research group of Ton Schumacher. His PhD research was funded by the Boehringer Ingelheim Fonds. At the moment, he works as a postdoc in Schumacher’s group. Schumacher calls Linnemann a ‘remarkably talented scientist’ who in his young career has already made important contributions to the development of cancer immunotherapy. During his PhD research Linnemann focused on helping T cells to recognize cancer cells. He did so by genetically modifying them to express new receptors, structures on the surface of T cells that can detect tumourspecific structures on the surface of cancer cells. Schumacher: “Carsten showed that providing new receptors to T cells can also cause problems. If the new receptors combine with receptors that were already present, they will recognize unexpected structures as ‘hostile’, with unexpected results. He also found a solution for this.”
Enterprising His work on T cells and receptor recognition earned Linnemann first authorship on two papers in Nature Medicine. Last December he
was again first author of a paper in Nature Medicine. This time, Linnemann helped to prove that not just CD8+ T cells, but also CD4+ T cells are able to recognize cancer cells as ‘hostile’ and ‘dangerous’. This makes these types of T cells into potential new targets for immunotherapy. Schumacher goes on by calling Linnemann “very passionate, enterprising, and an exceptional team player.” The enterprising part shows from the fact that Schumacher and Linnemann are currently founding a biotech company. The company will translate their basal scientific work on T cells and receptors into T cell therapies for cancer patients, in close collaboration with Schumacher’s group at the NKI.
Surprised “I was pretty surprised and very happy to hear that I’d receive this prize”, says Linnemann. “I have worked at the NKI for seven years now, and I have seen who won this prize before. They were all very good scientists. I feel so honoured to now belong to this row.” Linnemann plans to use the prize money of 6.000 euros on courses teaching strategies how to best evolve academic research into Biotech start-ups.
Mirjam Mol pleads for ‘high risk, high reward’ funding Unfortunately, the Netherlands have no big pharma development left after the demise of MSD’s R&D in Oss. Pivot Park director Mirjam Mol: “But we do have a lot of knowledge and expertise that could be brought to the point where it becomes attractive to big pharma. We should have a financing model to accommodate every step along this way. Yes, there is seed money, yes, there is venture capital, but for the ‘high risk, high reward’ part in between, the so-called ‘valley of death’, there is no solution. Policy is needed to tie the two loose ends together to make a strong chain.”
26-27 maart 2015 Lunteren Symposium 2015 Immunity and Science Fiction: the next 50 years in Immunology www.dutchsocietyimmunology.nl. 23 april 2015 Celkweek Hogeschool van Arnhem en Nijmegen 1 dag van 18.00-21.00 uur Locatie: Nijmegen Kosten € 537,00 Informatie en aanmelding: www.hanlifesciences.nl of E: email@example.com. 11-13 mei 2015 Jaarvergadering van de Association for Cancer Immunotherapy/CIMT The right patient for the right therapy Mainz, Duitsland 21 en 22 mei 2015 Workshop Analyse van Next Generation Sequencing (NGS) data Hogeschool Leiden, CBD Cursusprijs € 800,Inschrijven tot 7 mei 2015 Informatie & inschrijven: firstname.lastname@example.org. 27 mei 2015 Workshop NGS in de microbiologie diagnostiek Hogeschool Leiden, CBD Cursusprijs € 480,Inschrijven vóór 15 april 2015 Informatie & inschrijving: email@example.com 4 juni 2015 Scholingsdag Moleculaire Diagnostiek Hogeschool Leiden, CBD Cursusprijs: € 350,-. Inschrijven vóór 23 april 2015 Informatie & inschrijven: firstname.lastname@example.org 12 juni 2015 Masterclass Immunohistochemistry Hogeschool Leiden, CBD Cursusprijs € 480,-, bij inschrijven vóór 4 april 2015 € 435,-. Informatie & inschrijven: email@example.com 16 en 17 juni 2015 Workshop Primer en probes Hogeschool Leiden, CBD Cursusprijs € 740,-, bij inschrijven vóór 7 april € 675,-. Informatie & inschrijven: firstname.lastname@example.org.
Jesse Kraal In Immuun nr. 3 2014, on page 20 we erroneously contributed three photos to Jesse Klaver: the photos were made by Jesse Kraal.
29 juni – 1 juli Exploring Human Host-Microbiome Interactions in Health and Disease Wellcome Trust Genome Campus, Hinxton, Cambridge, UK 6-9 september 2015 ECI2015 Jaarcongres European Congress of Immunology www.eci-vienna2015.org 8 september 2015 (startdatum) Labmanagement - Communicatie Hogeschool van Arnhem en Nijmegen Vier wekelijkse bijeenkomsten op dinsdag 9.00-13.00 uur Locatie: Nijmegen Kosten € 795,00 Informatie en aanmelding: www.hanlifesciences.nl of E: info. email@example.com 8 september 2015 (startdatum) Labmanagement - Projectmanagement Hogeschool van Arnhem en Nijmegen Drie wekelijkse bijeenkomsten op dinsdag 14.00-18.00 uur Locatie: Nijmegen Kosten € 825,00 Informatie en aanmelding: www.hanlifesciences.nl of E: info. firstname.lastname@example.org 29 september 2015 (startdatum) Labmanagement - Kwaliteitszorg Hogeschool van Arnhem en Nijmegen Vier bijeenkomsten op dinsdag 14.00-17.00 uur Locatie: Nijmegen Kosten € 795,00 Informatie en aanmelding: www.hanlifesciences.nl of E: info. email@example.com 1 en 8 oktober 2015 ELISA theorie; achtergronden en kwaliteitsaspecten Cursusprijs € 720,- (bij inschrijven vóór 23 juli 2015 € 655,-) Hogeschool Leiden, CBD Informatie & inschrijving: firstname.lastname@example.org 2, 16 oktober, 6, 20, 27 november en 4 december 2015 Pathologie Hogeschool Leiden, CBD Cursusprijs € 1.950,- (bij inschrijven vóór 24 juli 2015 € 1.775,-) Informatie & inschrijving: email@example.com 24 november 2015 (startdatum) Immunologie Hogeschool van Arnhem en Nijmegen Vijf bijeenkomsten op dinsdag 18.00-21.00 uur Locatie: Amsterdam (Sanquin) Kosten € 552,00 Informatie en aanmelding: www.hanlifesciences.nl of E: info. firstname.lastname@example.org.
“Huge change of perspective” Scientific progress is, if anything, usually slow. Or isn’t it? In the case of cancer immunotherapy it certainly is not. The speed of development over the last four to five years, from concept and experiment to clinical reality, has been staggering. Professor Ton Schumacher of Antoni van Leeuwenhoek hospital gives an overview of what has been accomplished, what can be expected and which challenges remain. “In the USA, positive results have been reported with an experimental treatment of metastasized melanoma. The clinicians take T-cells from metastases, grow these in enormous amounts in the lab and place these cells back into the patient.” This quote on Tumour Infiltrating Lymphocyte (TIL) therapy is derived from an interview with Ton Schumacher in the publication ‘Immunologisch’ from 2011. Schumacher: “Back then, we thought about immunotherapy for cancer treatment as ‘possibly of value for treatment of melanoma’. Now we know that it is a valuable treatment option for quite a number of cancer types. Also, we think to have the insight for which types of cancer this will be the case.”
“With the ‘major DNA damage’ cancer types you can boost the existing immune reaction and with the ‘minor DNA damage’ types you can use gene therapy – with the hope that someday both approaches meet in the middle” Detection One of the keywords related to progress in cancer immunotherapy is detection. Schumacher: “Our immune system is primarily aimed at targeting invaders such as viruses and bacteria. The immune system recognizes these as notself because of their differing appearance. Most cancer types, however, are not caused by viruses, so how can the immune system recognize tumours? There are now many indications that the immune system is particularly good at detecting cancer types that are characterized by massive DNA damage.
Melanoma is an example, with DNA damage often caused by UV radiation exposure.” DNA damage causes proteins to be produced in cells that are normally not there. “These deviations don’t go unnoticed by the immune system. So with these types of cancer the immune system is in many individual cases capable of a first essential step: detection”, Schumacher concludes. His group is presently developing technology to measure whether an individual patient will develop an immune reaction against DNA damage or not.
Phase III Let us say the immune system has detected a tumour. Is it now also capable of destroying these alien structures? In many cases it isn’t, because a tumour deploys defence mechanisms that block this immune reaction. Insight in how tumours achieve that has grown in recent years. “Basically, the tumour cells present proteins on their surface which activate a ‘brake’ on immune cells”, says Schumacher. By now, a number of therapeutic options has been developed to overcome this inhibition. First of these is TIL therapy. In this approach not only the number of T-cells to attack the tumour is multiplied, the cells are also stimulated to be optimally active. Schumacher: “In a clinical trial led by professor John Haanen we have treated ten melanoma patients at the Antoni van Leeuwenhoek with TIL therapy. Five of them clearly benefitted from it. Two patients don’t even have any detectable tumour left. These results are in line with other studies. What’s more, with TIL therapy, resistance seems less of an issue than with, for instance, drugs that block signal transduction routes.” This first study is now followed up by a larger phase III comparative multicentre study in which the Antoni van Leeuwenhoek partners with Sanquin and centres in Great Britain and Denmark. In this study the results of TIL treatment of 150 patients will be compared to the results of patients treated with Ipilimumab, a drug designed to inactivate CTLA-4, one of the brakes on the immune system, allowing immune cells to attack a tumour. “It will be the first ever phase III study to compare TIL to a standard treatment”, says Schumacher.
Competitive field In the meantime, the field of drugs such as Ipilimumab that target brakes on the immune system, has become very competitive. A number of clinical trials is running, aimed at interfering with PD-1, a second brake in the immune system.
THEME: IMMUNOLOGY AND CANCER
These trials, which involve roughly halve the pharmaceutical top ten companies, seem to lead to an even stronger clinical response among an even higher percentage of patients than Ipilimumab. Schumacher: “These clinical responses are not solely limited to melanoma, but are now also seen in other types of cancer, in particular types of cancer associated with extensive DNA damage, such as lung cancer, bladder cancer, head and neck cancer and other types.” Furthermore, in addition to CTLA-4 and PD-1 there are many other brakes on the immune system. There is much research going on and there is still a lot to discover about these blockers and their possible role in other types of cancer. Many clinical studies are running which investigate the role of new drugs or the potential of drugs used in combination.
Patient specific When brakes on the immune system are blocked, this results in the activation of the entire immune system - and hence with the chance of side effects that resemble auto-immune disease. Schumacher and his group aim to find out in which patients the immune system is able to recognize a tumour as not-self and why that is. Schumacher: “If we know what an individual’s immune system can recognize as not-self on cancer cells, we can subsequently try to stimulate a specific immune reaction. We could push the immune system’s throttle, so to speak, to overcome the tumour’s braking action.” The DNA damage within a tumour is mostly specific for the tumour of that individual patient. This means that stimulation of an immune response against this should also be patient specific. In this way the treatment differs from the classic pharmaceutical approach. “The approach is new, but we know what process we need to develop. Although presently we don’t know for certain whether it will work. If successful, it will mean a major leap forward. A completely tumour specific approach would have the huge benefit of greatly reducing side-effects. That would be unique in cancer treatment”, Schumacher states.
Minor DNA damage Up to this point, it was all about cancer types that are characterized by large amounts of DNA damage. There are, however, also cancer types in which only a minor of DNA changes occurs, such as leukemia. In perspective: it ranges from a factor ten to a hundred less DNA mutations. Schumacher: “The consequence being that the immune system is in most cases unable to detect these cancer types by itself, at least not by the mutations they carry. Because of this, it is unsure as yet whether immunotherapy will be effective for these cancer types.”
Ton Schumacher: “By now we know for certain that immunotherapy is a valuable treatment option for quite a number of cancer types. Theoretically, immunotherapy could provide future treatment for all types of cancer.”
Changed perspective It already means a huge shift in perspective as compared to some years ago. “The world has changed”, Schumacher agrees. “We now witness major pharmaceutical companies investing massively in immunology based cancer drugs. We see how clinicians in all kinds of oncology specialisations embrace immune therapy. There is a lot of interaction with clinicians. All in all, the field has changed dramatically.” Among all this progress and optimism, one should bear the present limitations in mind. “At this point, we cannot tell for sure whether tumour specific immune stimulation is a feasible option. Moreover, we shouldn’t forget that many patients still have no response to the current immunotherapies”, Schumacher reminds. And then there is the matter that in some cases there is still only a temporary effect, before the tumour further evolves and creates resistance against the immune attack. “There is a lot of work to be done, clearly, but let’s not forget that we have only started the widespread testing of immunotherapy in the clinical setting a few years ago. The fact that resistance becomes an issue shows how far we’ve come in short notice. That’s the bright side of it.” Leendert van der Ent Photos Bureau Lorient Communicatie
Nederlandse samenvatting Immunotherapie tegen kanker:
Het perspectief is snel verbeterd
A number of research groups, including that of Schumacher therefore tries to change immune cells in such a way that they become able to detect these ‘minor DNA damage’ types of tumours too: “We know these tumour cells have specific proteins on their surface which normally don’t cause an immune reaction. Therefore we have to apply gene therapy to enable immune cells to recognize the tumour.” T-cells are isolated and activated in the lab. They are equipped with a receptor that can detect whatever has to be detected. Finally, the treated T-cells are reintroduced in the patient. Schumacher: “In the USA, a number of clinical studies aimed at treating hematologic malignancies with this approach have already been carried out. Preliminary results are quite spectacular.” This gives hope for the future. Schumacher further elaborates: “If these preliminary results can be sustained, this would imply that cancer immunotherapy can be applied to many types of cancer, both for the ‘major DNA damage’ as well as the ‘minor DNA damage’ tumour types. With the ‘major DNA damage’ cancer types you could boost the existing immune reaction and with the ‘minor DNA damage’ types you could use gene therapy – with hope that both approaches meet in the middle. This would enable the ultimate goal: treatment of the entire array of cancer types.”
In 2011 was immunotherapie tegen kanker nog grotendeels theorie en belofte. Professor Ton Schumacher van het Antoni van Leeuwenhoek: “Inmiddels weten we dat immunotherapie een waardevolle behandelingsoptie is bij verschillende soorten kanker. Ook het inzicht in de dynamiek van kanker in relatie tot het immuunsysteem is enorm gegroeid.” Bij kankertypen die gekarakteriseerd worden door een grote hoeveelheid DNA-schade, zoals melanoom en longkanker, herkent het immuunsysteem de tumor vaak als ‘vreemd’. Dat is nog niet hetzelfde als vernietiging, want tumoren hebben een verdedigingssysteem dat de immuunreactie afremt. Er zijn verschillende benaderingen om het immuunsysteem zodanig te versterken dat het over kan gaan tot de vernietiging van de tumor. Bij kankertypen die relatief weinig DNAschade aanrichten, zoals leukemie, herkent het immuunsysteem de kanker waarschijnlijk in de meeste gevallen niet. Gentherapie kan ervoor zorgen dat het immuunsysteem de kanker wel herkent. Als de voorlopige resultaten op dit gebied kunnen worden bevestigd, dan biedt dit de hoop dat op termijn voor vrijwel alle soorten kanker immunotherapie beschikbaar kan komen. Hoewel er nog een lange weg te gaan is, hebben de spectaculaire resultaten die de laatste jaren bereikt zijn al voor grote veranderingen gezorgd. De ‘big farma’ investeert nu veel in de ontwikkeling van op immunologie gebaseerde kankermedicijnen en clinici verwelkomen immunotherapie als nieuwe behandelingsoptie.
THEME: IMMUNOLOGY AND CANCER
A world’s first:
Preventive vaccine against hereditary colorectal cancer
Hoogerbrugge and De Vries: “What if we could boost the immune system of people with Lynch syndrome before cancer has got the chance to develop?” (photo Bureau Lorient Communicatie)
Jolanda de Vries and Nicoline Hoogerbrugge have vaccinated a group of twenty volunteers with Lynch syndrome against Lynch-related (hereditary) cancer at Radboudumc. They have mainly assessed the safety of the vaccine. This preventive vaccine against hereditary cancer should provide people with Lynch syndrome a more effective protection against the cancer for which they have a predisposition. The clinical trial is a first step in preventive vaccination against hereditary cancer. The Lynch syndrome stands for a hereditary mismatch in the repair genes. It causes a life time risk of up to 70% of developing colorectal cancer, against 5% in the general population. Furthermore, in Lynch syndrome cancer develops at a relatively young age. The immune system eradicates cancer cells continuously, especially in people with Lynch syndrome since their cancer cells and precursors thereof express antigens foreign to the immune system. Unfortunately, the balance between immune attack and tumour cell growth can be disturbed and tumours can grow. Jolanda de Vries (PhD) is professor at the Department of Tumor Immunology at the Radboud Institute for Molecular Life Sciences (RIMLS). She explains: “The body is very careful with DNA. DNA is built up in two matching strings. Normally, there is
a meticulous check on DNA that is copied. A DNA-repair system exerts it’s function when copying has not been perfect and repairs DNA-mismatches. When mismatches are not repaired, there is a higher probability of developing cancer precursor cells, derailed non-self cells, which can develop into cancer. That is what happens in people with Lynch syndrome: the repair system does not function flawlessly.”
No therapy available In Lynch syndrome patients, aberrant cells expressing incorrect proteins result from incorrect DNA translation. A distinguishing feature of these cells is that they present fragments of the incorrect protein, neo-peptides, on their surface. The immune
Who is Jolanda de Vries? Jolanda de Vries (PhD) is professor at the Department of Tumour Immunology at the Radboud Institute for Molecular Life Sciences (RIMLS). She was among the pioneers to translate dendritic cell biology into clinical applications. The first clinical phase I/II studies in which patients were vaccinated with DCs loaded with tumorspecific peptides were initiated in 1997. She also developed a novel immuno-monitoring assay that is highly predictive for extended survival after vaccination with DCs. Her primary scientific interest continues along the line of DC-immunotherapy and in particular the migration and imaging of DC.
system can recognise these neo-peptides as non-self. It is activated and destroys these cells before a tumour is formed. The immune system of people with Lynch syndrome works as it should and is activated. Their immune system is even capable of fighting the malignant cells. But in many Lynch individuals, the unusually large flood of non-self cells as compared to other people, overpowers their immune system and cancer develops. Nicoline Hoogerbrugge, MD, PhD, is internist-cancer geneticist and professor in Hereditary Cancer at the department of Human Genetics at the Radboudumc: “There is no therapy to prevent Lynch syndrome related cancer. Typically, we see adenoma/ polyps with an increasingly malignant nature occur in people with the syndrome. Our standard procedure is to track people with this syndrome from the age of 25 to offer them regular surveillance for adenomas. By performing colonoscopy and removing the adenomas we try to prevent colorectal cancer in an early stage.”
Dendritic cells turn the patient’s own T-cells against neo-peptides on (pre)cancer cells Prevention De Vries is specialized in the development of vaccines to boost the immune system in people with cancer. De Vries: “We witness cases in which immune therapy enables the immune system to beat cancer. Theoretically it is far more effective to boost the immune system before cancer has developed, when the patient’s immune system is still intact, than to try to beat an already existing tumour. But normally, preventive vaccines are not an option because you don’t know who is going to develop what kind of cancer and when. Hereditary cancer is the exception to this rule: with for instance Lynch syndrome you know exactly who runs a very high risk of getting colorectal cancer at an early age.” Hoogerbrugge: “Therefore Jolanda and I thought that it would be better to boost the immune system of these people before
De Vries: “The immune system reacts positively to the vaccine and we don’t notice an autoimmune reaction.”
cancer has got the chance to develop. What we typically observed in Lynch related colorectal cancer lesions was the presence of high numbers of immune cells compared to colorectal cancer lesions from non-hereditary cancer patients. Therefore we concluded that Lynch related cancers are immunogenic. So we thought: if only we could strengthen the immune reaction of these people in time, this would possibly enable them to withstand malignant cells.”
Building on experience Contrary to the situation with non-hereditary cancer, with Lynch syndrome it can be predicted which neo-antigenic peptides are presented on the surface of the cancer cells. De Vries: “This enables prevention in the same way as we normally perform immunotherapy against existing cancer, when there already is a response against tumour antigens. The vaccine consists of patients’ own dendritic cells loaded with neopeptides. The dendritic cells thus stimulate the patient’s own T-cells against these peptides; we don’t generate T-cells outside the body. In general, when you vaccinate against a mutation, this works far better than when you vaccinate against proteins
Who is Nicoline Hoogerbrugge? Nicoline Hoogerbrugge, MD, PhD is internist-cancer geneticist and professor in Hereditary Cancer at the department of Human Genetics at the Radboudumc. She is an adjunct clinical professor in hereditary cancer at Haukeland University in Bergen, Norway since 2011. Her clinical research is multidisciplinary in familial and hereditary colorectal cancer. Her expertise ranges from recognition of those at high risk for hereditary cancer, early diagnosis, innovation of care, new immunological treatment and cost efficacy studies. Her fundamental research is focused on identification and clinical characterization of novel genetic causes for colorectal and gastric cancer at adult age and in childhood.
THEME: IMMUNOLOGY AND CANCER
“There is no therapy to prevent the Lynch syndrome cancer”, Hoogerbrugge states.
that are also present on healthy cells. In this way, we hope to enable the T-cells to destroy the cancer. Or, alternatively in the case of preventive use, to destroy the precursors of tumour cells, or even to evoke an immune memory response, enabling the immune system to immediately recognise and destroy cancer precursor cells once they appear. In a therapeutic as well as in a preventive setting, we hope to alert the immune system against the threat, in order to stop the malignant process.” De Vries applies this kind of therapy in melanoma for eighteen years already. “We can build on this experience. The preventive vaccine benefits from progress made in cancer immunotherapy. We now make dendritic cells from precursor cells in the blood, which takes nine days. It has recently become possible to load natural dendritic cells from the blood with neo-peptides, which saves time and subsequently money. Moreover, in patients with cancer this is also more effective, so why not when applied preventively? The development is still too early and too specialised to perform a multicentre study yet, but this new method is more suited for such a study as it is technologically somewhat less demanding.” It is important to note that this preventive vaccination is totally unrelated to for instance mass vaccination against HPV. De Vries emphasises: “It is an individual approach, very laborious and costly, targeted at a limited group of people who run a high risk of developing cancer”.
Trial Trials aimed at prevention face very high demands, as people who are not ill may not be exposed to any kind of risk. Hoogerbrugge and De Vries were able to establish this, got approval for a trial and gathered a group of twenty still healthy volunteers with the Lynch syndrome to further prove the safety of their vaccine. This safety aspect especially regards autoimmune reaction. The vaccine rouses the immune reaction against the derailing intestinal cells before they have gotten the chance to develop into cancer cells. “It is important, that at the same time the immune balance stays intact; the immune reaction shouldn’t change in an auto-immune reaction.
The volunteers were vaccinated from spring 2014 onward. By now, all twenty volunteers have been vaccinated. “The first results are encouraging”, says Hoogerbrugge, “Although it is still too early to establish whether the immune system can indeed be boosted to prevent the development of colorectal cancer. ” “What we can say”, says De Vries, “is that the immune system reacts positively and that we don’t notice an auto-immune reaction. But it will take longer to draw conclusions on cancer prevention. We will only know that it worked, when people with Lynch syndrome develop colorectal cancer later than expected or even not at all.” If all goes well, it would also open perspectives for other types of hereditary cancer. Hoogerbrugge: “In that case we hope to administer preventive vaccines for other types of hereditary cancer as well. But for the moment we concentrate on this type of tumour. To find answers to the questions regarding the Lynch syndrome is difficult enough as it is.” Leendert van der Ent Photos Lagro Fotografie
Nederlandse samenvatting Eerste preventieve vaccinatie tegen erfelijke kanker Het Radboudumc heeft twintig vrijwilligers met Lynch syndroom, dat een kans van 70% op het ontwikkelen van erfelijke darmkanker geeft, gevaccineerd om kanker te voorkomen. Het afweersysteem is niet altijd krachtig genoeg om kankercellen, ontspoorde cellen, helemaal op te ruimen. Bij mensen met het Lynch syndroom, die drager zijn van een erfelijke fout in één van de DNAreparatiegenen, ontstaan vaker “foute” cellen. Nicoline Hoogerbrugge, hoogleraar Erfelijke Kanker in het Radboudumc: “Voor het Lynch syndroom bestaat nog geen therapie. Mensen met dit syndroom screenen we vanaf vijfentwintigjarige leeftijd regelmatig op adenomen, het voorstadium van darmkanker. Door adenomen te verwijderen proberen we het ontstaan van darmkanker vroegtijdig te voorkomen.” Het preventieve vaccin tegen erfelijke darmkanker moet hen een nog effectievere bescherming bieden. Hoogleraar Translationele Tumorimmunologie in het Radboudumc Jolanda de Vries ontwikkelde dit vaccin: “De vrijwilligers die in ons onderzoek het vaccin krijgen, hebben nog geen kanker en we hopen met het vaccin het ontstaan van darmkanker te voorkomen.” Het brengt de afweer tegen ontsporende darmcellen op gang voordat deze zich ontwikkelen tot kankercellen. Daarbij moet wel een balans bewaard blijven; afweer mag niet omslaan in een autoimmuunreactie. De eerste resultaten zijn hoopgevend. “Inmiddels zijn alle mensen gevaccineerd”, aldus De Vries. “We zien een goede reactie van het afweersysteem. Het geeft hoop dat het vaccin werkt. Zekerheid hebben we pas na langere tijd, als deze mensen veel later dan gebruikelijk of zelfs helemaal geen darmkanker ontwikkelen.”
Circulating Tumour Cells:
A key to cancer detection, drug Leon Terstappen brought the Circulating Tumour Cell capture technology CellSearch to the clinic. It allows patient monitoring and early detection of metastases instead of only risk-profile based prognosis. Further development of the technology, for instance in the European CTC-TRAP project, enables cell-specific information to direct personalized, targeted therapy at an early stage of (recurrent) cancer. Only ten percent of cancer patients die from their primary tumour. Ninety percent succumb as a consequence of metastases caused by cells that have detached from the primary tumour. These Circulating Tumour Cells (CTC) move through the blood. Most of these cells are destroyed in the liver or by the immune system, but some escape to form new malignancies/ metastases. If only these CTC could be detected.
That was the completely new hypothesis behind the research started by professor Leon Terstappen MD, PhD in the USA in 1995. Terstappen indeed found a way to detect CTC. This was the start of a hundred million dollar research project and the development of Immunicon Corporation’s CellSearch technology. The technology was taken over by Johnson & Johnson in 2008, after it had been FDA cleared and brought to the clinic in 2004. CellSearch is now deployed for patient monitoring in about four hundred hospitals worldwide, one of which is the Erasmus Medical Centre in Rotterdam.
Leon Terstappen: “In drug development we wish to make the step from stating that a drug responds to a percentage of tumour cells with a certain receptor to the percentage of patients who actually respond to the therapy.”
“Normally, you will need tumour cells to make an assessment of which therapies will be the most effective to arrive at a personalised approach”, says Terstappen. “But it is hard to take a biopsy from tumour metastasis. It is of course possible to store cells from the primary tumour after surgery for later use, but these may no longer be representative of the tumour at the time therapy is needed. Cancer cells mutate, change their phenotype and genotype; this is exactly what makes these cells resistant to therapy. The most critical aspect in cancers is their heterogeneity and constant change. Cells from the primary tumour may therefore not be representative for the metastasis. You need up-to-date tumour information.” So how do you know what to look for? Terstappen: “My hypothesis was based on the fact that epithelial cells don’t belong in the blood. When you find those, they have to come from a
THEME: IMMUNOLOGY AND CANCER
development and treatment carcinoma. To find these, you take an immunologic marker, antibodies that target epithelial cells to label them. You connect the marker to small magnetic particles. Subsequently it becomes possible to extract the magnetically labelled CTC from the blood with a magnet.” This is the simplified story of the CellSearch technology. The scientific version was published in the New England Journal of Medicine and in Clinical Cancer Research in 2004. To find CTCs is much harder than suggested above, given the fact that CTCs in the blood are extremely rare. To find CTCs, the procedure starts by using 7.5 millilitres of blood. In a first step, billions of red blood cells and millions of white cells are separated from the rest. What remains are the CTC among now hundreds to thousands of white blood cells. The CTC can be found with fluorescently labelled antibodies to distinguish them from the remaining white blood cells. Terstappen underlines: “Immunology is vital to this technology. Everything revolves around the antigens present on the CTC, antibodies to these immunologic markers are labelled with magnetic particles to select them from the blood and fluorescent markers to enable their identification under the microscope.”
Prognosis to observation The first hypothesis was proven: yes, these cells can be detected. The second hypothesis was, that detection of these cells point out the need for further treatment. This was also clinically proven from 2004 onward, first with breast cancer and later with prostate and colon cancer: finding CTC means a worse prognosis for the patient. Terstappen: “When CTC are still present after three weeks of therapy, it means the therapy hasn’t worked and something else has to be undertaken to battle the cancer.” Conventionally, once the primary tumour is removed from cancer patients, they are treated according to a prognostic risk profile that is based on the findings around the time of surgery and the characteristics of the tumour at that time. Terstappen: “The CellSearch technology enables the step from therapy based on risk for recurrence to the actual observation of the presence of micro or macro metastasis.”
Same cells, different receptors Since 2004, the number of scientific publications on the CellSearch principle has skyrocketed to 10,000 to 30,000 at the moment. These articles focus on further development and deployment of the approach in new applications. Terstappen:
Everything revolves around the antigens present on the CTC “Apart from the diagnostic monitoring application – to establish whether recent therapy has been effective – there is another major field of application. Once you have detected the CTC, it is also attractive to find out to what substances they are sensitive. A lot of studies therefore involve pharma trials in which CellSearch is used to determine whether a new target is able to bind to the CTC.” Unfortunately, this is also more complex than previously perceived. Terstappen: “The biological Herceptin, for instance, is approved for aiming at the Herceptin receptor on the cells of the primary tumour. When the CTC, the tumour cells in the blood, are investigated at the individual cell level, it appears that there is a large variation of the expression of the receptor on the tumour cells implying that the therapy can only be effective on a portion of the tumour cells. This is vital information that was not taken into account before.” “This might mean that a patient may need additional drugs that can attack the other tumour cells in order for the therapy to be
Who is Leon Terstappen? Leon Terstappen received his Medical Degree from the University of Groningen in 1983 and his PhD from the Department of Applied Physics of Twente University in 1988. He worked on stem cell characterization as a post-doctoral fellow and in various research positions at the research institute of Becton Dickinson, San Jose, CA, USA. He joined Immunicon Corporation (now J&J) in 1994 to pioneer the detection of tumour cells in blood of cancer patients as Chief Scientific Officer and Senior Vice President of Research and Development. He developed the CellSearch-technology for which the Prix Galien was received in 2009. In 2007 he was appointed Professor of Medical Cell Biophysics at the University of Twente and at the UT MIRA-Institute for Technical Medicine and Biomedical Technology. The International Society for Analytical Cytology (ISAC) awarded him the ‘Max Fulwyler Award for Innovative Excellence’ for his research into cell detection and analysis in 2012. Terstappen has about fifty patents to his name and is involved in the start-up Vycap.
effective. Good future biologicals will need to target a relevant combination of receptors”, Terstappen concludes.
Immunology is vital to this technology More detail How unfortunate this complexity may be, it is highly relevant information for drug development as well as further patient treatment in the clinic. Terstappen: “Traditionally, tumour cells are characterized as -, +, ++ or +++ for the presence of potential therapy targets. The development of CTC detection is aimed at a much more detailed and quantitative description of the tumour profile, at the genetic expression as well as the protein expression level. Various receptors occur in a certain measure on the CTCs. This receptor-profile, which informs on the per cell expression of different receptors, should define treatment. It will for instance make clear whether targets for new treatments should be aimed at the oestrogen receptor or the androgen receptor regardless of the origin of the tumour.” More knowledge and more detail result in a better hit chance for new targets – the most pressing challenge for the pharmaceutical industry. This is the philosophy behind the European precompetitive Public Private Partnership Innovative Medicines Initiative (IMI) CANCER-ID, in which Terstappen coleads a 14 million euro project. Terstappen: “We wish to validate technologies for blood-based biomarkers to determine the absence or presence of drug targets and assess the response to treatment. We will be able to establish which therapy is the most effective for the individual patients. This program will enable
Immune defense live at work Terstappen: “What you see here is immune defense live at work: a first image of a Circulating Tumour Cell that we detected, with T-lymphocytes attached to it. It would be a great leap forward if we understand what actually happens. The immune system obviously recognizes the CTC as foe and tries to destroy it. In cancer, this ultimately fails. The challenge is to multiply the destructive power of the T-cells. We can make is visible. I hope that researchers are willing to take this mechanism from there and determine what actually happens. For instance: what do T-cells recognize in these tumour cells? Why can they in many cases destroy cancer cells, but in some cases not? Are there not enough T-cells? Are they not powerful enough? How can they be strengthened? How often does it occur that one or more T-cells are attached to CTC? What happens next – destruction or failure? What are the differences between individuals? This has never been systematically investigated.”
pharmaceutical companies to deploy the best technology in their studies for new drugs and accelerate the process of getting the drugs only to the patients that will benefit from it.
Therapy? Apart from supporting diagnosis and enabling new drugs research, the merits of the CTC detection technology to provide a therapy are now under investigation in the six million euro European Research project CTC Circulating Tumour Cells TheRapeutic Apheresis (CTC-TRAP). Terstappen explains: “This new project aims at real time liquid biopsy in patients suspected of metastasis. Instead of taking only milliliters of blood, this project involves the patient’s total blood volume. We strive to filter the blood in a way comparable to dialysis. In half an hour, all the patient’s blood passes the filter, leaving the CTC behind.” Filtering five liters of blood takes the technology to a whole new level as compared to analysis of a couple of millilitres. Terstappen: “All the CTC have to be detected for analysis in order to get all the info needed for tailored treatment for individual patients. The challenge is huge. Once CTC-TRAP is developed, it also has to be validated. Validation of the technology in lung cancer is the aim of CANCER-ID another upcoming European project which involves 26 centers throughout Europe. The ultimate goal is to personalize therapy on the basis of tumour cells available during the entire disease process. In this way you can adapt therapy according to new information. CTC contain this information, so we need to get as much out of it as possible. Hopefully we will at some point be able to treat patients based on their CTC phenotype and genotype.” Leendert van der Ent Photos Bureau Lorient Communicatie
THEME: IMMUNOLOGY AND CANCER
Treatment of cancer patients with immunotherapy looks promising. Sanquin Reagents notices this too. There is a strong interest in her technologies to monitor antigen-specific T cell populations in clinical trials with new medication.
Relevant epitope iden�ﬁca�on
Neo-‐epitopes Vaccine targets Proof of Mechanism Treatment op�misa�on Candidate selec�on
Monitoring Cytotoxic T cell popula�ons
Immunotherapy & Immune monitoring tools Powered by patent families WO 2006/080837, WO 2010/060439, WO 2012/076708
Monitoring Immunotherapy Immunotherapy focuses on stimulating cytotoxic T cells to eliminate cancer cells. How do you know which target is important and do T cells respond to this? Sanquin has extensive expertise in immunology and blood cells and has, among others, developed technology aimed at both identifying relevant targets and phenotyping a wide range of antigen-specific cytotoxic T cells (CTL).
however feasible to analyse up to 10 antigens simultaneously, and we are happy to discuss licensing options with interested parties. We have already been contacted by several Dutch and international biotechnology and pharmaceutical companies.”
Immune monitoring CTL populations: Proof of mechanism “In contrast to other assays, our assays allow you to discriminate whether an activation observed is non-specific or is related to anticipated antigen-specific effects of treatment. In addition, we can determine whether the antigen-specific population consists of predominantly effector, effector memory, central memory or naïve T cells.” Astrid Visser explains. “This allows researchers to verify at an early stage in a clinical trial whether their therapy does what it is expected to do. It gives insight into the effects of an experimental therapy and can be used in the process of establishing a proof of mechanism.”
Improved epitope selection One of our proprietary toolsets improves the selection of relevant T-cell epitopes. When you know which proteins are relevant in a tumour, in silico programs can identify candidate epitope peptides that are likely to bind MHC complexes. We found that even though some of these programs work well, they are not suitable for ranking the best candidates. Sanquin’s peptide-MHC binding assay selects and confirms the best candidates presented by the MHC (eliminating about 20% of predicted candidate epitopes that do not actually bind), and also provides a ranking that is well linked to in-vivo relevance (leading to more optimal immunotherapy vaccines).
As servicetest or for licensing Sanquin Reagents offers this patented technology, developed together with the Netherlands Cancer Institute, as one of its services in the field of immunotherapy. “Antigen-specific immune monitoring is fairly complicated; execution requires a certain level of expertise. For most experienced FACS users it is
This article was made possible by Sanquin Reagents. Contact: Astrid Visser, Business Development Manager, email@example.com, +31 (0) 20 512 32 40, http://www.sanquin.nl/reagents
Lia van der Hoek wins Immuno Lia van der Hoek (AMC) has won the Immuno Valley Award 2014 for her pitch ‘Virus discovery, fast, sensitive and cheap’ at the Immuno Valley Annual Conference 2014. Van der Hoek, associate professor at the Academic Medical Center, developed a technology that enables fast identification of viruses. She already discovered around fifteen new viruses. “In virus discovery things go so fast that in five years time all human viruses may have been identified.”
Viruses are curious things. They’re generally not considered living organisms, since they’re just strains of DNA or RNA in a protein coat (and in some cases an envelope of lipids). They replicate inside the living cells of other organisms, from animal, humans and plants to microorganisms. So far, about 5,000 human and animal viruses have been described in detail. But there are many more species and virologist Lia van der Hoek has developed a technology to discover these. That is easier said than done, since it is difficult to separate the DNA or RNA of a virus from the host’s DNA and RNA. Only a fraction of a clinical sample is relevant for sequencing, but how to purify the fraction to focus on?
A nice hobby Lia van der Hoek got her PhD in 1998, working on HIV-1 in the digestive tract. “In 2001, I started to develop my own method that would make it possible to discover viruses in a fast and sensitive manner”, she says. At the time it was sort of a hobby project, executed next to her regular work. A very successful hobby project that is, since it resulted in 2003 in the VIDISCA (Virus Discovery cDNA-AFLP) technology. Her first shot at a sample containing an unidentified virus was a hit: she discovered a new human coronavirus (HCoV-NL63) with her brand new technology. For the next five years, she shifted her
Valley Award 2014 Winning pitch at Immuno Valley Annual Conference Lia van der Hoek (AMC) presented her pitch ‘Virus Discovery, fast sensitive and cheap’ at the Immuno Valley Annual Conference. There were six presenters, who all did a great job, according to jury members Peter van Dijken (TNO), Mark Offerhaus (Micreos) and William Weldon (Elanco Animal Health). Van der Hoek however won the Immuno Valley Award 2014. The jury was impressed with her work, her presentation and the practical application of her idea. Van der Hoek received the award and a cheque of 2500 euros. Van der Hoek: “I also got the opportunity to participate in a two day ‘pitch training’. The training taught me a lot on how to present an idea and captivate the attention of possible partners. I can also put these skills to good use in the classroom!” Van der Hoek praises Immuno Valley for building bridges between scientists and businesses, in human and animal health. “Immuno Valley makes it possible for me to come into contact with people whom I would otherwise not meet, thus creating relevant opportunities for partnerships. I hope the Immuno Valley Award will be another incentive for partners to explore possibilities for cooperation that are mutually beneficial.” More information on Immuno Valley at www.immunovalley.nl
attention towards this novel virus until 2008 when she took up further improvement of the virus discovery method. She did so by combining VIDISCA with Next Generation Sequencing and novel purification techniques. Next Generation Sequencing provides ten thousands to millions of sequences per sample, an extremely powerful tool which is today introduced in many genomics projects.
cheaper, costs less computing power, is faster and enables analyzing more samples.” For this achievement she received the Immuno Valley Award 2014 (see box).
A new virus a new pathogen? Van der Hoek is continuously improving her method. “I’m always on the lookout for ways to make VIDISCA more sensitive and faster”, she says. But the method is already a success. So far, it has enabled Van der Hoek to discover around fifteen new viruses, eight of which have been published. What happens after discovery? “Then we always determine whether an infection with the novel virus is associated with disease. We make a diagnostic assay and find out whether it presents itself more often in sick patients than in a healthy controls. If this association is confirmed the virus is a serious pathogen candidate. The next step is to fulfill the Koch’s postulates: culture the virus from a sick animal or human, applying pure virus in an animal model and isolate the virus from these guinea pigs. If the animals get the disease then the causal relationship between infection and disease has been validated”.
Modern times The revolutionary Next Generation Sequencing technology has facilitated identification of unknown viruses at a speed that was out of reach some years back. “In virus discovery things happen so fast it’s almost unbelievable”, Van der Hoek says. “I can’t even exclude the possibility that five years from now all human viruses will be identified. After that, of course, we still have a massive challenge in linking these viruses to diseases.” Alinda Wolthuis Photo Immuno Valley
Lia van der Hoek winnaar Immuno
Eliminate the enemy
Valley Award 2014
One of the main competitors for virus discovery is ribosomal RNA that is massively present in clinical material and competes with sequencing of viral genomes. The purification technique that Van der Hoek introduced enables separation of the relevant part from the irrelevant part of a sample. By applying special elongation and non-elongation oligonucleotide primers ribosomal RNA is removed from a sample. “The fact that we remove a large part of the irrelevant material means that we have less sequencing to do: it’s
De pitch ‘Snel, gevoelig en goedkoop virussen ontdekken’ tijdens de Immuno Valley jaarconferentie 2014 leverde Lia van der Hoek de Immuno Valley Award op. Van der Hoek, Universitair Hoofddocent op het AMC, ontwikkelde een technologie die een snelle identificatie van virussen binnen bereik brengt. Ze ontdekte verschillende nieuwe virussen in de afgelopen jaren. “In virusdiscovery gaan de ontwikkelingen zo snel dat binnen vijf jaar alle humane virussen geïdentificeerd zouden kunnen zijn.”
Onvermoede relatie tussen schiz Bij het ontstaan van schizofrenie spelen erfelijke factoren een grote rol. ‘Dat weten we uit tweelingonderzoek’, zegt Eske Derks die zich als hoogleraar Genetica vooral richt op psychiatrische aandoeningen. ‘Heeft een eeneiige tweelingbroer of -zus schizofrenie, dan is de kans vijftig procent dat de ander het ook krijgt. Bij de overige broers of zussen daalt die kans naar zo’n tien procent, maar dat is nog altijd aanzienlijk hoger dan gemiddeld.’
Ook omgevingsfactoren spelen een rol. Kinderen die bijvoorbeeld in de winter of het vroege voorjaar worden geboren, hebben een licht verhoogde kans op schizofrenie. ‘Dat zou te maken kunnen hebben met een grotere kans op infecties in het najaar’, zegt Derks. ‘Infecties tijdens de zwangerschapsfase hebben mogelijk een negatief effect op de (hersen)ontwikkeling van de ongeboren vrucht.’
Genetische kwetsbaarheid Er zijn meer aanwijzingen dat infecties, immuniteit en afweer iets te maken kunnen hebben met schizofrenie, en andersom.
Derks: ‘Mensen met schizofrenie zijn vatbaarder voor infecties. Niet alleen voordat de diagnose schizofrenie wordt gesteld, maar ook daarna. Recent is duidelijk geworden dat medicatie die de afweer dempt bij een deel van de schizofreniepatiënten een goed effect heeft op de ziekte. Als aspirine wordt toegevoegd aan de gebruikelijke anti-psychotica verbetert de toestand van sommige patiënten.’ Derks en haar collega’s onderzochten daarom of er een genetische overlap bestaat tussen schizofrenie en immunologische aandoeningen. Met andere woorden: zijn genen of genetische gebieden die bij schizofrenie betrokken zijn, óók actief bij het ontstaan van immunologische aandoeningen, zoals diabetes type 1, de ziekte van Crohn en reumatoïde artritis (RA)? Voor dit onderzoek bekeek Derks de genetische variatie in grote patiëntengroepen. ‘Er zijn nog maar weinig genen bekend die de kans op schizofrenie vergroten’, zegt Derks, ‘maar met statistisch onderzoek kunnen we ook naar wat grotere stukken DNA, ofwel genetische regio’s, kijken waarvan we vermoeden dat dergelijke genen zich daar moeten bevinden. Die aanpak hebben we gevolgd voor vier patiëntengroepen: mensen met schizofrenie en mensen met de ziekte van Crohn, diabetes type 1 en reumatoïde artritis.’
‘Beruchte’ regio In de analyse voor schizofrenie springt bijvoorbeeld een ‘beruchte’ regio op chromosoom 5 in het oog. Daar liggen diverse genen die al eerder met schizofrenie in verband zijn gebracht, zoals ACSL6 en NEUROG1. Maar – en dat maakt het interessant – diezelfde regio bevat ook veel genen die een rol spelen in de afweer. Er zitten bijvoorbeeld enkele genen die coderen voor interleukines, dat zijn signaalstoffen die delen van de afweer aansturen. Op chromosoom 6 ligt ook zo’n gebied met zowel immunologische genen (Humane Leukocyt Antigenen, HLA) als genen die een grotere kans op schizofrenie geven.’ Derks: ‘Ons onderzoek, gepubliceerd in Schizophrenia Research, wijst uit dat er een duidelijke genetische overlap bestaat tussen schizofrenie enerzijds en de drie immuunziekten anderzijds. Vrij snel na onze publicatie verscheen een artikel in Nature met nog grotere patiëntengroepen, maar met een vergelijkbare conclusie. De volgende fase van het onderzoek zal zich moeten richten op de biologische signaalroutes van de aandoeningen, op de moleculaire onderbouwing van de ziektes. We moeten nu de stap maken van statistische verbanden naar biologische verklaringen.’
Bijsturen Dergelijk onderzoek vergt een lange adem en kan zowel vanuit de psychiatrie als de immunologie worden gevoed. Met interessante perspectieven, denkt Derks. ‘Stel dat we inderdaad kunnen
Science across borders
ofrenie en afweerstoornissen aantonen dat ontstekingen en bepaalde variaties in de afweer de kans op schizofrenie vergroten. Dan kunnen we mensen met een hoog risico op schizofrenie misschien wel preventief met bepaalde ontstekingsremmers gaan behandelen. Juist omdat schizofrenie sterk genetisch is bepaald, kunnen we potentiële
patiënten relatief eenvoudig opsporen. Iemand beter maken die schizofrenie heeft, is moeilijk. Preventie door tijdig bij te sturen, is bij deze ziekte een optie die veel meer kansen biedt.’ Pieter Lomans
Remarkable correlation between immunology and schizophrenia Genes that make people vulnerable for schizophrenia are partly also involved in immunological diseases such as type 1 diabetes, Crohn’s disease and rheumatoid arthritis. The next step is to find out what this partial overlap of the schizophrenia related genes with the immunological disorder related genes tells us. Hereditary factors play a vital role in the occurrence of schizophrenia. “We know that from research with twins’, says professor in Genetics Eske Derks, who mainly focuses at psychiatric diseases. ‘When an identical twin brother or sister has schizophrenia, the risk is fifty percent that the other also gets this disease. Among the other brothers and sisters this chance drops to about ten percent, which is still far higher than average.” Environmental factors also play a role. For instance, children born in winter or early spring run a slightly higher risk of getting schizophrenia. “This could have to do with a higher risk of infections during the autumn”, says Derks. ‘Infections during pregnancy possibly have a negative effect on the brain development of the unborn child.’
Genetic vulnerability There are more indications that infections and immunity on one side and schizophrenia on the other side are interrelated. Derks: “Schizophrenia patients are more susceptible to infections. This not only goes before the diagnosis schizophrenia is made, but also after. Recently it has become clear that immune moderating medication has a dampening effect on the manifestation of schizophrenia among a part of the patients.
Notorious region In the analysis of schizophrenia a ‘notorious’ region on chromosome 5 stands out. Derks: “Various genes are located there which have been identified as related to schizophrenia, such as ACSL6 and NEUROG1. But – and that is what makes it interesting – this very same region also contains many genes that play a role in the immune system. There are for instance some genes which code for interleukins, signal substances which can activate part of the immune system. On chromosome 6 there is also a location with ‘immunological’ genes (Human Leukocyte Antigens, HLA) as well as genes that cause a higher risk of schizophrenia.” Derks proceeds: “Our research results, published in Schizophrenia Research, point out that there is a clear genetic overlap between schizophrenia on the one side and type 1 diabetes, Crohn’s and rheumatoid arthritis on the other. Already quite quickly after our publication was published, an article was published in Nature on the basis of even larger patient groups, with a comparable conclusion. The next phase of the research will have to focus on the biological signal routes of the diseases, on their molecular basis and process. We now have to make the step from statistic correlations to biologic explanations.” Pieter Lomans This article was previously published in AMC Magazine December 2014 / January 2015
Cancer research at the Pivot Park Pivot Park is the science park in Oss aiming at bridging life sciences and technology. The park has come a long way since its official opening in 2012. Thanks to its roots in MSD and predecessor Organon - a company that cherished immunology as a spearhead – cancer immunology research is very well represented on the site. Companies such as BioNovion and Acerta Pharma are living proof of this. Activities on the Pivot Park are more or less equally divided into one third new product development, one third pharmaceutical contract research and manufacturing and one third enabling services. About two thirds of the new product development involves immunology research. A considerable part of this is dedicated to cancer research, with links to the NKI, academic medical centers and large pharmaceutical companies. Chief Scientific Officer Andrea van Elsas of BioNovion, a pioneer at the park: “The Netherlands is home to a disproportionately large, globally recognised expertise in immunology- not in the least in cancer immunotherapy. Organon was the corporate figurehead in this respect, a position that could be maintained under Schering-Plough and MSD.” Pivot Park emerged from major MSD reorganizations and former MSD-experts started their own spin-outs at the park, just like BioNovion. “It is no surprise that immunology is a major research field here”, Van Elsas concludes.
There is a large variety of research and development activities. These range from lead discovery, development, compound manufacturing from lab scale up to pilot plant production and early stage clinical trials to dossier submission. This covers the middle of the pharmaceutical value chain, the bridge between academia and big pharma. There are many connections between the companies on the site. Mol: “We actively stimulate collaboration through our open access labs and community events.” Mol sees the transformation of Pivot Park into a lively campus as a dot on the horizon. “The shared entrance with MSD and Aspen necessitates closed gates. A separate entrance would enable evolution towards an open campus, with the look and feel of a community with its own restaurant and an outdoor café.”
Highly valuable equipment “Pivot Park is ready for its next step, to become a vibrant life sciences community”, Van Eenennaam, Chief Operating Officer of BioNovion agrees. BioNovion started three years ago in an incubator setting and moved last December to a larger facility of its own, also at the Pivot Park. Van Eenennaam: “Thanks to equipment and labs that were already in place, we could start experimenting from the start. We have our own facilities now, but still do use the park’s infrastructure.”
Mirjam Mol: “We offer the R&D infrastructure of large companies to SMEs” Dot on the horizon “After two and a half years, Pivot Park now houses some 35 companies with about 350 employees”, says park director Mirjam Mol. “Fifty percent of the available capacity is now rented out. Pivot Park has state-of-the-art core facilities and infrastructure for pharmaceutical research and development, with special start-up packages, ranging from simple glassware to advanced equipment such as HPLC systems and parallel synthesis equipment. But the park also hosts manufacturing capabilities, has the Pivot Park Screening Center for high throughput screening and provides all-in lab services, from handling complex gases to waste management. Companies can buy a subscription to lab equipment or bring their samples to have them analysed or screened. We offer the R&D infrastructure of large companies to Small and Mediumsized Enterprises (SMEs).”
Number one on the wish list: to open the gates to welcome visitors to the campus. (Photo Pivot Park)
IMMUNOLOGY AND CANCER
Acerta Pharma joined Pivot Park more recently. Executive vice president discovery Allard Kaptein: “It is a huge benefit to be able to use the highly valuable equipment of the Screening Center. Another opportunity was that as a result of the latest MSD reorganization in July 2014 we could hire experienced researchers, who could immediately contribute to our activities.” Acerta Pharma is an R&D company focused on oncology and auto-immune disease. Kaptein: “It’s all about activated immune cells gone out of control, which can lead to auto-immune disease as well as rapid malignant cell growth. Our specialization is the development of covalent binding drugs, which attach permanently to target proteins. Once the drug is bound its presence in the circulation is no longer needed. While still being effective on the target, it lowers the risk of off-target adverse effects.”
Hans van Eenennaam: “Pivot Park is now ready for this next step, to further grow into a vibrant life sciences community” From idea to clinical trial BioNovion builds on academic knowledge of antibody application against the immune checkpoint protein CTLA-4 and on Organon’s research into therapeutic antibodies against another immune checkpoint protein, PD-1 (see the interview
Other companies on the Pivot Park involved in cancer immunology research: • Contract research company NTRC also has its own line of research in oncology and immunology. • Synaffix is active in the field of Antibody Drug Conjugates (ADC). • Glycostem is dedicated to innovative (stem) cell therapy against diseases, including cancer. • Intervention in the process of degenerative diseases such as diabetes and cancer is the aim of Lead Pharma.
with Ton Schumacher). This Organon work, started in 2004, eventually led to Pembrolizumab as the first anti PD-1 drug approved by the FDA – Merck’s Keytruda. Van Eenennaam: “Our place in the value chain is to bring research ideas towards clinical trials. This proof of concept work is based on fundamental understanding of tumour immmunotherapy. Our technology platform with its Organonroots enables us to identify and produce antibodies which activate or inhibit immune cells in order to optimally mobilize them against cancer – or other immune relate diseases.” BioNovion CSO Andrea van Elsas: “We do not produce just antibodies, we make ready-for-testing, functional, producible and safe antibodies. This is a big difference, that enables the vital step towards a marketable product.”
Allard Kaptein: “It is a huge benefit to be able to use the highly valuable equipment of the Screening Center” BioNovion strives to combine these competences with academic knowledge and therefore seeks collaboration with academic researchers who dispose of a biologic model system. Van Eenennaam: “This combined knowledge can actually get new drugs to the clinic.Take for instance our collaboration with Jannie Borst at the NKI, who brings in twenty years of knowledge of CD27. This has, with the support of TI Pharma, now led to a new product concept.” Another collaboration involves professor Kenneth Anderson at the Dana-Farber Cancer Institute in Boston, regarding treatment of multiple myeloma. In addition to growth by successful start-ups, Pivot Park also manages to attract parties from outside. Last year’s decision by the clinical trial services multinational Quintiles to join Pivot Park meant a breakthrough to the international scene. Mol is confident that a transformation towards an open campus will persuade more companies to join. “This would bring eighty to ninety percent occupation within reach.” Leendert van der Ent This article was made possible by Pivot Park and Brabantse Ontwikkelings Maatschappij (BOM), www.bom.nl.
Breakthroughs at the intersection “The single most important thing is passion”, says immunologist Hidde Ploegh. Passion for research brought him to the other side of the Atlantic. Already as a graduate student he worked at Jack Strominger’s lab at Harvard. Today, he heads the Ploegh Lab at the Whitehead Institute for Biomedical Research. He studies the various tactics that viruses deploy to evade our immune responses and the ways in which our immune system distinguishes friend from foe.“In my lab I’m surrounded by (bio)chemists, cell biologists and engineers. It’s a good place to be.” His career has linked him to a number of respected institutions: Harvard University, the University of Cologne, The Netherlands Cancer Institute (NKI), the VU University Amsterdam, Harvard Medical School, The Whitehead Institute for Biomedical Research and Massachusetts Institute of Technology. Since 2005 Ploegh is on the faculty at the Whitehead Institute and MIT. Here he is in his element. “It’s a curiosity-driven environment and I’m at liberty to follow my research instincts to my heart’s content.”
Fascination for the MHC Whereas there may have been changes in his employers, Ploegh has been consistent in his research object of choice: the major histocompatibility complex, MHC. His first achievement was cloning a cDNA for a major MHC antigen. He continued to study
The joy of teaching “I like to teach. I get the impression that in the Netherlands many professors are reluctant to enter a classroom, they prefer doing their own thing in the lab. At MIT we all are expected to teach, I myself just finished a 24 x 2 hour course and I had tremendous fun in the process. We have a fascinating profession and I enjoy talking about it. I don’t make use of powerpoints, I use old-fashioned chalk and blackboard, because that makes it easier for students to follow the pace of exposition. I also make sure to tell things they can’t find in their textbooks - and then test it in the exam.”
synthesis and trafficking of MHC products, but broadened the scope by studying pathogens that hide by incapacitating MHC products and the cellular protein degradation pathways that provide them with their peptide antigens. At the Whitehead Institute, Ploegh studies how viruses evade immune responses, and the ways in which the immune system distinguishes friend from foe. The Ploegh Lab reported new mechanisms by which dendritic cells detect the presence of antigens and instruct the immune response, using Class II MHC-eGFP knockin mice and live cell imaging. Ploegh also helped to elucidate how products of the major histocompatibility complex are assembled and are delivered to the right destination to help an immune response kick in: herpes viruses such as HCMV evade the immune system by selective destruction of Class I MHC products.
New labeling methods
Nobel Prizes “The number of immunologists to have won a Nobel Prize, is surprisingly large. That can be explained by the fact that discoveries in immunology are very recognizable. The same goes for genetics, by the way.”
The Ploegh Lab applies chemistry-based strategies to illuminate aspects of protein quality control. “At the Whitehead I find myself surrounded by really good chemists and specialists from other disciplines. That suits me like a glove: working at the intersection of disciplines can lead to real breakthroughs. I myself probably know more chemistry than the average immunologist and I almost certainly know a lot more about immunology than the average chemist. That enables me to define interesting research subjects. My colleagues are always willing to lend me a hand to solve technical problems.”
of disciplines Extremely charismatic, phenomenal classes “I was inspired by Jon van Rood, an extremely charismatic immunologist who was well ahead of his time. I also credit Jan Drenth, biochemist at Rijksuniversiteit Groningen. His classes were phenomenal: he was a credit to biochemistry as well as eloquent, and he had a marvelous sense of humor.”
Dutch immunology stands out “The Netherlands stand out in immunology. Every single Dutch university boasts a credible effort in immunology and the Dutch Society for Immunology is a large and vibrant community. There is a downside to this: objective peer review of research results is more difficult because all Dutch immunologists know each other. They should look abroad for peers.”
So far, Ploegh and his coworkers have emphasized the generation of chemical tools to study proteasome activity and the roles of lysosomal and ubiquitin-specific proteases. A recent innovation is the generation of cloned mice with lymphocytes specific for pathogens such as Toxoplasma, influenza and herpes viruses, using the technique of somatic cell nuclear transfer. As one of the keynote speakers at he NVVI Winter School 2014, Ploegh impressed the audience with his lecture on new labeling methods that enable labeling any protein in mice, thus disclosing chains of events that remained invisible until now. “With our new method, we can see whether immune cells can find a tumour and if so, what they do to it. We can follow this process in time. This might help us to determine why immunotherapy is successful in some cases and unsuccessful in other cases. Developing this method required a combination of immunological, chemical and biochemical knowledge, all of which is present at the Whitehead.”
Breaking new ground Ploegh, who is a contributor to over five hundred papers, always opts for fundamental research. “Applied research is just not my cup of tea. I’m better at exploring new grounds, to do what has not been done before and to develop new technologies, new approaches and new tools. I’m happy to leave applied science to
others. I encourage people to build on our results and use our discoveries to their own ends. I regard it as a compliment.” He is horrified by the current drive for valorisation. “I don’t understand the attraction of it. In the Netherlands it is presented as an American example to follow, whereas in reality in the States such a concept or philosophy doesn’t exist. Let’s be clear on this: fundamental research is the source from which spring most if not all medical applications such as new chemotherapeutics or biologicals. If the fundamental research is sound, applications will follow. One shouldn’t be forced into ‘valorisation’.” So far, all of his ideas have landed somewhere. “Sometimes in places I hadn’t expected, but that’s all the more fun.”
The second immunology revolution “In the fifties, immunology was responsible for the elimination of nearly all infectious diseases in children. Now, immunology is about to revolutionize healthcare for the second time: cancer immunology will prove to be a breakthrough. In malignant melanoma, forty percent of patients can be treated successfully with antibodies. Soon this treatment option will become available for other cancers as well.”
PhD students have mastered this and then graduate, they unfortunately leave the club.”
Level playing field Over the years, Ploegh has seen come and go many coworkers. A significant number of them are now leading scientists in the Netherlands, for instance Sjaak Neefjes, Ton Schumacher, Huib Ovaa, Peter Peters, Madelon Maurice, Marianne Boes and Hermen Overkleeft. “It’s great to see how they’ve developed their own line of research and become leading scientists in their own fields. I like to think that the work they did with me has contributed to their current achievements, has helped them to identify new opportunities and to challenge themselves. I like to keep in touch, too.” Cross-Atlantic relations are therefore warm and frequent, as is emphasized by his part-time appointment as professor in cellular protein chemistry at Utrecht University. How does Ploegh regard the current scientific climate in Europe? A decade ago, he stated in an interview that the climate in the US was more ‘vibrant and lively’ than in most places in Europe and praised himself lucky to be in the Boston academic climate, with its ample resources and intellectual firepower. That may hold truth to some degree even today, he says, but the differences have become less apparent since he first set foot in America in 1992. “There has been a leveling of the playing field on both sides of the Atlantic. Email and social media make information exchange quick and simple. Universities have access to all top publications. That has promoted interaction across the Atlantic - and stimulated science on both sides.” Alinda Wolthuis Photos Bureau Lorient Communicatie
“With our new method we can see whether immune cells can find a tumour and, if so, what they do to it. We can follow the process in time. This might help us to determine why immunotherapy is successful in some cases and unsuccessful in other cases.”
The explorer Ploegh likes breaking new ground. He does not have a long-term strategy and he doesn’t know what he’ll be doing five years from now. Ploegh: “There is stability in the sense that there is longterm financing for my program, but that’s about it. Flexibility is key at the Whitehead. That is both my wish as it is the institute’s policy: I do not employ staff nor analytical researchers on a permanent basis. PhD students enlist for the MIT graduate program, not for a defined project. They can’t get a permanent position with us. There is a downside to this flexibility, of course. Along with the graduating PhD students, their specialized knowledge is at risk of disappearing. Also, certain technologies, such as the generation of cloned mice, requires expertise that takes time to master. But once
Passie voor vernieuwing “Het allerbelangrijkste is passie”, zegt immunoloog Hidde Ploegh. Passie voor onderzoek bracht hem naar de andere kant van de Atlantische oceaan, waar hij leiding geeft aan het Ploegh Lab bij het Whitehead Institute for Biomedical Research. Hij bestudeert de taktieken waarmee virussen een immuunrespons weten te omzeilen en de manier waarop het immuunsysteem vriend en vijand uit elkaar houdt. Zijn focus ligt op fundamenteel onderzoek. Dat hij in het Whitehead voornamelijk nietimmunologen om zich heen heeft, bevalt hem uitstekend: “Voor vernieuwing moet je op het kruispunt van disciplines zijn.”
Cancer Vaccine Tracking project:
Translating immunology to the clinic The CTMM Cancer Vaccine Tracking project has brought expertise of chemistry, pharmacology, imaging and immunology together. Thanks to this multidisciplinary project, a therapeutic vaccine against cervical and vulvar cancer now approaches market introduction. “The project plays a vital role in bringing ‘molecule’ to ‘man’, bringing scientific knowledge to the clinic”, prof.dr. Ferry Ossendorp, LUMC and prof.dr. Kees Melief, CSO at ISA Pharmaceuticals in Leiden conclude. Some decades ago it was discovered that T-cells can recognize virus infected cells and tumours via the presentation of foreign proteins presented as short peptides on the cell surface. This knowledge opened the way towards immunotherapy against cancer. “We have proceeded on that”, says prof. dr. Ferry Ossendorp of the Leiden UMC, principal investigator of the CTMM Cancer Vaccine Tracking project. “On the basis of the immune system’s ability to recognize foreign peptides we developed an immunotherapy based on therapeutic peptide vaccination against virus-induced tumours, in particular against the viral onco-proteins of the Human Papilloma Virus that can cause cervical and vulvar malignancies and head and neck cancer.”
The right peptides The heart of the matter is, to teach the immune system to elicit a strong response against the right proteins. Some researchers choose to work with short peptides, that are loaded directly onto all kinds of cells by Human Leukocyte Antigen (HLA) molecules. “This introduces the risk of an unbalanced immune response. We therefore choose to work with long peptides Kees Melief and Ferry Ossendorp: “We hope to learn how the synthetic long peptide therapies behave in-vivo, which would help us to direct our research with optimal efficiency” (Photo Bureau Lorient Communicatie)
instead. Long peptides require a processing step, which only dendritic cells can accomplish. To stimulate a T-cell response with a low dosage of long peptides in the proper stimulatory context of a DC is a safer and more efficacious solution. No active drug is introduced, but a pro-drug that is metabolized to an active drug in the DC”, says Melief. Ossendorp: “Furthermore, long peptides don’t cause HLA matching problems and can thus be indiscriminately used in all patients. Nine of the long peptides cover the E6 oncogene protein, four cover the E7 oncogene protein. The vaccine is injected subcutaneously with a small needle to the skin of the upper arm. The vaccinated peptides gain easy access to the many local dendritic cells (DCs). These move to lymph nodes, where they activate both types of T-cells, CD4 helper cells and CD8 killer cells. Subsequently, the activated T-cells go out to turn against the E6 and E7 proteins specifically expressed by the (pre-) malignant tissues.
Preliminary clinical results ISA Pharmaceutical sponsors a clinical trial with 48 patients in nine centres in the Benelux. Clinical results show that synthetic
- - - - -
Leiden University (Organic Chemistry group Hermen Overkleeft - fluorophore labeling, TLR ligand conjugate, Synthetic Large Peptide development) Leiden UMC (Tumor immunology, In vivo imaging. GMP Facility: SLP vaccine production, fluorophore peptide production. Hospital: clinical trial) Utrecht University (nanoparticle formulation development) Percuros BV (imaging) ISA Pharmaceuticals BV (vaccine development, clinical trials)
Over the past seven years, the Center for Translational Molecular Medicine (CTMM) established a highly effective infrastructure for public-private partnerships, translating science into better healthcare. Newly developed molecular technologies in the field of diagnostics and imaging make early diagnosis and patient-tailored treatment possible for oncologic, cardiovascular and neurodegenerative conditions, as well as for infectious and auto-immune diseases. CTMM and TI Pharma are preparing to merge into one organization to collaborate on public-private healthcare innovation.
long peptide immunotherapy is safe and effective in premalignant indications. Melief: “In combination with standard chemotherapy it looks favourable in patients with late stage cancer. In the pre-clinical as well as the clinical model, chemo plus long peptides works better than checkpoint blocking plus long peptides. We show that chemotherapy leads to depletion of immunosuppressive cells, resulting in a synergistic effect with long peptide treatment.”
“In this trial, the long peptides are directly connected to immune stimulating molecules: Toll Like Receptor (TLR) ligand conjugates. In pre-clinical models this new approach behaves more bioactive than the unconjugated long peptide version, because it targets the peptide to the DC and simultaneously activates them. Ossendorp explains: “It thus stimulates important DC functions more effectively and consequently also induces a stronger T-cell response at lower doses.” The multidisciplinary approach of the CTMM Cancer Vaccine Tracking project, combining imaging, synthesis and immunology, brought this progress about. In a previous project, TI Pharma supported the all-important intellectual property positioning and drug development. CTMM subsequently sponsored the step from molecule to the clinic. Vital support, according to Melief: “Thanks to this, large patient groups could eventually benefit from these specific immunotherapeutics. That’s how it was intended. It is good to see that it really works out that way.” Ossendorp: “The project has brought it within our reach to offer the best possible immunotherapy to the patient.”
Imaging gives insight The Cancer Vaccine Tracking project focuses on in-vivo imaging of the therapeutic from the injection site to the draining lymph nodes, in order to offer the researchers maximum insight in the immunization process. Prof.dr. Hermen Overkleeft’s organic chemistry group connected a near infrared fluorophore label to one of the long peptides. This, together with Percuros’ imaging technology, enables non-radioactive imaging deep into the tissue. Melief: “The synthetic nature of this project’s Synthetic Long Peptides (SLP®) and the in-vivo imaging aspects enable replacement of the traditional empirical approaches in research by step-by-step design, in which pharmaco kinetics and pharmaco dynamics are taken into account, including time, place, formulation and dosage aspects.” This year, a trial with healthy test persons will start at the Center for Human Drug Research (CHDR) to test the imaging results of the fluorophore-labelled SLP. Also a new formulation is tested, where the emulsion is replaced by a nanoparticle encapsulation slow release alternative. This delivery technology was developed by the Utrecht University Pharmacology Group of prof.dr. Wim Hennink. “We hope to learn how the long peptides behave invivo, which would help us to direct our research with optimal efficiency”, says Ossendorp.
Next generation Apart from the labelling and formulation, the T-cell response to the vaccine itself is also improved. A clinical trial with new generation synthetic long peptides will start shortly. Melief:
Nederlandse samenvatting Het CTMM-project Cancer Vaccine Tracking brengt chemische, beeldvormende, farmacologische en immunologische expertise samen om een therapeutisch vaccin tegen baarmoederhals- en schaamlipkanker naar de markt te brengen. De ontwikkelde beeldvormende mogelijkheden verschaffen inzicht dat in het vaccinonderzoek de stap van trial en error naar meer gerichte en geplande ontwikkeling mogelijk maakt. “Het project is een belangrijk voorbeeld van hoe je van ‘molecuul’ naar ‘mens’ gaat en wetenschappelijke kennis in de kliniek brengt”, concluderen prof. dr. Ferry Ossendorp, LUMC en prof. dr. Kees Melief, CSO van ISA Pharmaceuticals in Leiden. Dit artikel is mogelijk gemaakt door CTMM: www.ctmm.nl
Immunity and Science Fiction:
The next fifty years of On the 26th and 27th of March 2015, the Lunteren Symposium will take place. It is dedicated to the 50th anniversary of the Dutch Society for Immunology (NVVI). ‘Immunology research in the next fifty years’ was chosen as a fitting team for this spring meeting. What immunological challenges lie ahead? Nationally and internationally renowned speakers comment on this. In the opening session Hans Clevers (Hubrecht Institute, Utrecht) will kick off with a presentation on ‘Stem cells: Dr Jekyll or Mr Hyde?’ Clevers is the President of the Royal Netherlands Academy of Arts and Sciences (KNAW), professor in molecular genetics and one of the world’s leading researchers on stem cells
and their potential for regenerative therapy. Clevers was the first to identify stem cells in the intestine. He discovered similarities between the normal renewal of cells in the intestine and the onset of colon cancer. His group has since been able to grow mini-organs or ‘organoids’ from
immunology research individual stem cells of the intestine. These epithelial organoid cultures are genetically and phenotypically extremely stable, allowing transplantation of the cultured offspring of a single stem cell, as well as allowing disease modelling by growing organoids directly from diseased patient tissues. These exciting developments have great potential to repair genetic defects in stem cells of patient tissues. The second speaker, Nienke Vrisekoop (University Medical Center Utrecht), is inspired by immune cell dynamics, specifically T-cell dynamics in time and space. Immune cells are the most mobile cells in the body. They can ‘patrol’ through tissues and reach inflammatory sites quickly. To study T-cell dynamics in living mice she uses intravital 2-photon microscopy. Vrisekoop also studies the relationship between the binding strength of the T-cell receptor - the unique receptor which T-cells use to recognize their specific antigen - to presented foreign antigen and the strength of self-peptide Major Histocompatibility Complex (MHC) reactivity. Mathematical modelling will enable to estimate the true immune cell contact times and will enable to predict how the observed contacts add up over time and space.
T-cells Christopher Love (Associate Professor of Chemical Engineering at the Massachusetts Institute of Technology (MIT) and Ton Schumacher (professor of Immune Technology at the Leiden University Medical Center, with his lab at the Netherlands Cancer Institute (NKI) Amsterdam) are the speakers of the session on T-cells. They will reveal novel approaches to dissect heterogeneity within T-cell populations and tell how they use this to manipulate the molecular processes underlying differentiation of and immune recognition by T lymphocytes. The main aim of Love’s research is to understand how heterogeneity in cell populations affects their collective behaviours as a system. Research in The Love Lab combines ideas from materials science and interfacial chemistry to enable new micro- and nanotechnologies for studying the biology of complex cell collections in a quantitative manner. The lab’s researchers have developed new processes for analysing large numbers of individual living cells quantitatively and dynamically, with the use of high-throughput screening of monoclonal antibodies, micro tools for profiling immune responses and microfluidic systems for live cell imaging. The resulting profiles will enable understanding of for example the precise cellular signatures that characterize an immune response to one disease state or another.
Schumacher aims to design and test novel concepts for adoptive immunotherapy of cancer. To this end he designs novel technologies that can be used to examine and modify antigen-specific T-cell immunity. He also uses these tools to unravel and manipulate the molecular processes underlying immune recognition by T lymphocytes. The lab has developed methods to trace cell fate at the single cell level, using genetic barcodes. Another line of research involves mapping of all tumour-specific mutations within human cancer lesions, using exome sequencing, which could serve as regression antigens in patients. The link between cancer genomics and T-cell immunology may suggest novel ways to harness the T-cell repertoire in human disease.
B-cells Hergen Spits (Academic Medical Center Amsterdam) and Michael Reth (scientific director of the BIOSS Centre for Biological Signalling Studies, Freiburg, and full professor in Molecular Immunology at the University of Freiburg) represent two research extremes in the field of B-cell biology. This field is about the discovery of therapeutic antibodies for clinical use and the development of techniques for a profound study of signalling mechanisms in B-cells, respectively. Apart from professor of Immunology, Spits is also co-founder of AIMM Therapeutics, a company that aims to discover therapeutic antibodies for the treatment of cancer and infectious diseases. This company’s approach is based on genetic programming of human memory B-cells into long living plasmablasts. The method provides a tool to immortalize B-cells, enabling the rapid generation of high-affinity human monoclonal antibodies without the need to use extensive molecular engineering techniques. Another strategy involves the development of bispecific antibodies to create a covalently linked IgG antibody heterodimer. Reth’s research line focuses on the Nano-scale organization of the B-cell receptor (BCR) on resting and activated B-cells. The exact organization of proteins in the membrane of living cells is still poorly understood. By combining novel assays to detect the proximity of two target proteins and through state-of-the-art super-resolution microscopy techniques, Freiburg researchers have been able to investigate the structure, organization and dynamics of the BCR and its interaction with co-receptors. The studies suggest that many membrane proteins are not freely diffusing monomers, but rather multicomponent protein complexes pre-organized in nano-size protein islands.
Extracellular vesicles The evening lecture will be dedicated to extracellular vesicles (EVs) or, by their former name, exosomes. Clotilde Thery is Research Director and group lead at the Institute Curie in Paris. Already in 1998 she was fascinated by EVs. These are formed in the endocytic compartment and subsequently secreted outside the cell. Although EVs can be produced by almost all cells, Thery is especially interested in those produced by dendritic cells (DCs) and tumour cells. By using extensive proteomic analysis she has studied the composition of EVs and provided a detailed map of the proteins selectively targeted to these vesicles. Using state-ofthe-art cell biological approaches, the intracellular mechanism of EV formation and secretion has been analysed, which will provide novel therapeutic options in the fight against cancer.
High resolution In the session on high resolution, Daniel Davis (director of research in the Manchester Collaborative Centre for Inflammation Research (CCIR), professor at the University of Manchester), and Paul Parren (Senior Vice President and Scientific Director of Genmab in Utrecht and professor at the University of Southern Denmark (Odense) will demonstrate what is experimentally possible with new imaging techniques that dive into the nano-range. Davis specializes in visualizing cell-cell interactions, including the immunological synapse, a special organization of molecules between immune cells that is important for the activation of T-cells and the killing of virally infected cells or tumour cells. Scientists at Davis’ lab discovered that long, thin connections, ‘membrane nanotubes’, developed between cells. Cells use these to communicate at a distance. Using high resolution microscopy, Davis’ team demonstrated that immune cells transport genetic material, small RNA molecules, through these membrane nanotubes to cancer cells. Thereby they inhibit the proliferation of these cells. Davis will focus on the possibilities and limitations of high- and superresolution microscopy to demonstrate which questions can be addressed with this promising new approach. Parren is highly interested in the biology of antibodies and in particular how these immune molecules can be applied successfully for immunotherapy. To understand how antibodies behave inside the body, Parren and his team explore the structure of antibodies and how they exert their effector function. He uses high resolution mass spectrometry to study the molecular details of an antibody, including the sugar structures. He will discuss various novel high resolution approaches that enable investigating in great detail both the structure and function of proteins in general and antibodies in particular.
Neuronal control “Influencing the immune system with your brain, wouldn’t that be great?”, asked both Peter Pickkers and Paul-Peter Tak themselves a number of years ago. Since then they have investigated this idea and exploited their findings in the clinic
for the benefit of patients. In the session on neuronal control they will demonstrate that there is considerable interplay between the autonomic nervous system and the immune system: both the parasympathetic (via the vagus nerve) and the sympathetic nervous system exert regulatory functions on the immune response. The so-called ‘cholinergic anti-inflammatory pathway’ may represent a novel therapeutic modality to limit inflammation in various conditions. As professor of Experimental Intensive Care Medicine at the Radboud University Medical Center Nijmegen, Pickkers investigates the immune system during sepsis. In his presentation he will focus on the parasympathetic cholinergic anti-inflammatory pathway and will discuss interactions between the sympathetic nervous system and the immune system. He will demonstrate that through practicing techniques developed by ‘iceman’ Wim Hof it is possible to activate the sympathetic nervous system at will and that this results in suppression of immune responses. Paul-Peter Tak is Senior Vice President / Head of the Immuno Inflammation Unit at GlaxoSmithKline (Stevenage, UK) and professor of Rheumatology at the University of Cambridge. He has a strong background in successfully developing novel treatment options for patients with rheumatoid arthritis and other chronic inflammatory diseases, in particular vagus nerve stimulation as a new bioelectronics anti-inflammatory approach. In his presentation he will not only present the results of this revolutionary work, but also expand upon the potential use of bio-electronics in the treatment of other diseases in the near future.
Advanced imaging The closing lecture ‘Intravital immunology: seeing T-cells make decisions’ will be delivered by dr. Thorsten Mempel, principal investigator at the Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital in Charlestown MA, and associate professor of Medicine at Harvard Medical School in Boston MA. Mempel’s lab seeks to understand how the function of T-cells is regulated through their interaction with other cells, structural tissue components and soluble mediators that they encounter in tissues. For this purpose the lab researchers use advanced imaging techniques such as multiphoton intravital microscopy (MP-IVM). This allows them not only to observe the migration of immune cells in their physiological tissue context in living humanized mice, but also to monitor the activities of various signaling pathways and how these are regulated by intercellular contacts.
NVVI Symposium Lunteren, 2015, March 26-27 Congrescentrum De Werelt, Lunteren, The Netherlands Thursday March 26
Friday March 27
Welcome and Introduction
â€˜Meet-the-speakerâ€™ breakfast sessions
Session 1: Perspective: In vitro vs In vivo Chair: Annemiek van Spriel
Session 4: High resolution Chair: Martijn Nolte
Hans Clevers (Utrecht, The Netherlands)
Dan Davis (Manchester, UK)
Nienke Vrisekoop (Utrecht, The Netherlands)
Paul Parren (Utrecht, The Netherlands)
Session 2: T cells Chair: Debbie van Baarle
Session 5: Neuronal control Chair: Sander Tas
Christopher Love (Cambridge, USA)
Peter Pickkers (Nijmegen, The Netherlands)
Ton Schumacher (Amsterdam, The Netherlands)
Paul-Peter Tak (Stevenage, UK)
Session 3: B Cells Chair: Rudi Hendriks
Closing lecture Chair: Maaike Ressing 13.15
Hergen Spits (Amsterdam, The Netherlands)
Michael Reth (Freiburg, Germany)
Drinks and Dinner
Thorsten Mempel (Charlestown, USA )
Evening lecture Chair: Esther de Jong 20.00 Clotilde Thery (Paris, France) 21.00
EU project COMPARE:
Supporting the global fight against infectious diseases The new 20 million euro EU project COMPARE in the Horizon 2020 program aims to reduce the impact and cost of infectious disease outbreaks. To attain this goal, COMPARE strives to speed up worldwide detection of and response to infectious disease outbreaks, both in human and in veterinarian settings. New genome technology should enable this. A consortium on 28 institutes led by Erasmus MC Rotterdam and the National Food Institute of the Technical University of Denmark Kopenhagen sees to the project’s execution. Other Dutch consortium partners are iBMG (The Institute of Health Policy and Management) of Erasmus University Rotterdam, the AMC in Amsterdam, RIVM and Artemis Wildlife Health BV.
Human and animal health are under constant and increasing threat of disease outbreaks around the globe caused by microorganisms such as bacteria and viruses. Diseases are transmitted by foods or the intervention of other vectors, such as mosquitos and midges. New outbreaks occur and epidemics recur. Apart from the grieve caused by mortality and disease, societal and economic impact of these incidents and disasters can be huge. The pressure on healthcare can be immense, production of livestock suffers, confidence in food security and supply is hampered and trade and economy can come to a standstill. Swift action can see to it that outbreaks are contained and their consequences remain relatively limited. The West-African Ebola-crisis unfortunately proves that such swift action is not common practice yet. The recent avian influenza outbreak equally demonstrates that a pathogen can have a major international impact.
Real time global exchange Predominant to limiting the consequences of outbreaks is being able to quickly identify the microorganisms behind it. Within the COMPARE framework, a global platform will be developed by using new techniques for whole genome sequencing of pathogenic microorganisms. This should result in a globally accessible databank. The data should be exchanged in real-time to relate them to existing insight about the mechanisms behind disease. The databank should therefore also store knowledge on the properties of pathogens, including the mechanisms these use and the transmissibility of the viruses and bacteria between humans and animals. In addition, information on treatment methods and infection prevention will be included. Complex analyzes needed to examine the data will be provided in an automated manner, so that even laboratories without this expertise will henceforth be able to use it.
The development of such a system must be carried out carefully, to prevent wrong conclusions and wrongful warning calls. This is why physicians, diagnostic laboratories, patient groups, governments, the food industry and other businesses will be closely involved in the development of the system.
One Health approach Frank Møller Aarestrup from the Technical University of Denmark, joint leader of the European consortium with Prof. Marion Koopmans, head of Virology at Erasmus MC: “The aim is that the platform can be used to harmonise the way scientists, authorities, doctors and organisations around the world collect samples, generate genome sequencing data and carry out risk assessments. This new approach to disease surveillance will be able to revolutionise the way we combat diseases globally.” Zoonoses – diseases that can spread from animals and food to humans – are the cause of many epidemics internationally. For this reason COMPARE is based on a collaboration across sectors and land borders and builds on the One Health approach. The consortium unites doctors, veterinarians, virologists, and microbiologists to develop the platform in collaboration with epidemiologists and bioinformaticians. Koopmans: “The databank information can be compared to all sorts of other available information, such as clinical and epidemiological data, in a way that is accessible to physicians, veterinarians and other end users. This will enable us to detect new infectious disease outbreaks more rapidly and in a better manner and to make better decisions on appropriate measures to be taken to prevent further spread and on the best treatment options for affected patients.” Contact: firstname.lastname@example.org
IMMuno VAllEy ConsortIuM
Immuno Valley: connecting human and animal health Immuno Valley is a business driven public-private consortium at the interface of human and animal health. Immuno Valley’s highly qualified scientific and business partners collaborate in order to translate R&D expertise into new products for the diagnosis, prevention and treatment of infectious diseases. Academia, businesses and SMEs active in the field of human and animal health are welcome to join the Immuno Valley consortium of almost 40 scientific and business partners. Immuno Valley partners benefit from:
overview of experts and their expertise
• Collaboration opportunities: academic and
including research facilities, in the field of
business partners find each other during matchmaking events or are approached 1 to 1. In case a new call opens or other
infection and immunity in human and animals. • Event discounts: free or discounted tickets
funding opportunities arise, partners
for Immuno Valley Annual Conference,
interested in a certain topic are gathered for
theme-related symposia and matchmaking
joint proposal writing or advised on how to proceed. • Matchmaking: account contacts (twice per year) on business opportunities and
events. • Open communication channel: facilities to publish your scientific or business case on Immuno Valley’s website.
actualities resulting in customized advise and professional support in finding suitable partners and accessing new funds. • Subsidy-alerts: Immuno Valley provides up-to-date and personalized mailings about new funding opportunities. • Immuno Valley Expertise Database: Access to the online expertise database offering an
More information W www.immunovalley.nl E email@example.com P +31 30 2531142 Mijke Vogels (accountmanager)
The Art of Saving a Life Edward Jenner’s Smallpox Discovery © Artwork by Alexia Sinclair
Set in an 18th century English doctor’s surgery, this stunning portrait features Dr. Edward Jenner inoculating James Phipps, the first person to receive the smallpox vaccine. Dr. Jenner’s pioneering work in the late 18th century led to the eradication of smallpox in 1980. Alexia created and photographed the entire tableau. The aristocratic woman in the center represents how smallpox did not discriminate, affecting the rich and poor alike. The many flowers throughout the piece symbolize the global impact of smallpox, and the skulls on every bottle the ephemeral nature of life and death. This portrait by Alexia Sinclair is part of the ‘Art of Saving a Life’-collection commissioned by the Bill & Melinda Gates Foundation. More than 30 world-renowned photographers, painters, sculptors, writers, filmmakers, and musicians tell the stories behind the success and the future promise of immunization. Stories of risk and bravery, passion and dedication of scientists, the love of parents, and the determination of health workers. Hear, see and feel the tremendous impact of immunization!