This summer, 40 research projects into the coronavirus pandemic and the consequences of thisstarted with funding from the Ministry of Health, Welfare and Sport. We rapidly need a lot of knowledge about the SARS-CoV-2 virus and the disease COVID-19. Animal-free models can play a role in that because the results can be better translated to humans and yield outcomes faster. Five projects with animal-free research will therefore start.
In April of this year, the Ministry of Health, Welfare and Sport made 40 million euros available to ZonMw and NWO for research into the coronavirus pandemic and the consequences of this for society, such as the illness COVID-19. The research results should become available as quickly as possible for everybody who has an interest in this illness: doctors and nurses, national and municipal governments, care providers and citizens. With animal-free innovations, research can be done better and faster. And that is vitally important now: the committee of the ZonMw research programme More Knowledge with Fewer Animals therefore made extra funding available to realise projects with such animal-free innovations within the research programme COVID-19. ‘I’m proud of the performance delivered by everyone involved, applicants and assessors, because an awful lot of work was realised in a short space of time’, says Dick Tommel, chair of the committee More Knowledge with Fewer Animals.
The Dutch Society for the Replacement of Animal Testing also made funding available for this call for proposals. ‘Many animal experiments are performed for research into COVID-19. We will almost certainly see that in the figures’, says Debby Weijers of the Society. ‘With this initiative, we want to contribute to COVID-19 research without animal experiments. For this disease in particular – and similar diseases in the future – we hope to achieve a lot with human models. That is because these models reveal far more accurately how the disease progresses in humans. Also, with human models, we can personalise the research, for example by using stem cells or computer simulations based on patient data. With such approaches, we can better investigate why the virus makes certain groups more ill than others. Animal models will not be able to provide an answer to these and other questions.’
In this special round, embedded in the COVID-19 call for proposals, five projects were selected that make broader use of existing animal-free innovation or make the development of new animal-free innovations possible.
Theo Geijtenbeek, Professor of Molecular and Cellular Immunology at Amsterdam UMC, will lead research into the possible preventive effect of the anticoagulant low molecular weight heparin against SARS-CoV-2. Already, COVID-19 patients are now immediately administered heparin by means of injections when they are hospitalised in order to prevent blood clots. But Geijtenbeek and his group discovered that this drug also blocks the binding of the virus to cells and consequently prevents infection. Now they want to investigate whether inhaling heparin can have a preventive effect so that, for example, healthcare personnel can use a heparin inhaler to protect themselves from infection. The first step in the research is innovative, says a proud Geijtenbeek. Volunteers will be asked to inhale heparin via the nose. Subsequently, the researchers will remove some nasal mucosa cells (just like during a coronavirus test) and will subsequently expose these cells to the virus to investigate the antiviral effect of heparin. Geijtenbeek: ‘We want to do it in this way to prevent the need for animal experiments and so that we can enter the clinical phase earlier. And time is an important factor in this pandemic’. In addition, the research group will make use of a dynamic human cell model to further investigate the effect of heparin on the coronavirus infection.
A dynamic cell model is a research setup that the consortium led by Robbert Rottier, senior researcher at the Pulmonology Department of the Sophia Children’s Hospital, will use for their research. Static lung systems are used a lot in today’s coronavirus research. The disadvantage of these models is that they only simulate the functioning of human lung cells to a limited extent. Therefore, Rottier’s team, together with professor Roman Truckenmüller from Maastricht University and the MERLN Institute, will produce a closed dynamic system. They will use an existing bioreactor to simultaneously culture human cells from both the epithelium of both the respiratory passages and blood vessels. By transporting microfluids through this, a dynamic system will arise. That can be used to better study the development and progression of our infections such as COVID-19. Through the collaboration with Truckenmüller, an expert in the area of nanotechnology and biochips, and the MERLN institute, this system can also be rapidly produced at a commercial scale and made available to laboratories. The team is also working on a protocol so that the model can be used without the need for extra training. According to Rottier, working on animal-free innovations has additional advantages: ‘Since we started working on this type of innovation, we have increasingly looked for alternatives within the research group, and our use of laboratory animals has decreased.’
Working on animal-free innovations can make researchers more aware of how they do their research. It is also what Jeffrey Beekman, Professor of Cellular Disease Models at University Medical Center Utrecht discovered. ‘Before this, I did not particularly focus on animal-free research, but the call for proposals and writing the proposal has made me more aware of this. I now pay more attention to the materials I work with, such as the human cells that originate from individuals and the serum that I use to allow those cells to grow. That serum often has an animal origin.’ For research into COVID-19, Beekman and his research team will use cell models to study how the coronavirus infection works in vivo on various organs: upper respiratory passages (nose), lower respiratory passages (lungs), intestines and kidneys. ‘By combining these models with unique genetic characteristics of the cell donors, you can compare the different tissues and discover factors that influence the effects of the coronavirus and the efficacy of the drugs used against it’, says Beekman. ‘In the case of COVID-19, that is particularly important because the virus affects various organs.’
How can you simulate a COVID-19 patient so that you can investigate why some patients develop blood clots? That was the question which Andries van der Meer, Associate Professor in Applied Stem Cell Technology at the University of Twente, and his team will tackle. Some 10-30% of people with COVID-19 who are hospitalised, end up developing blood clots. Consequently, this group of patients has a far worse prognosis. The basis for the project is a model of mini-blood vessels on a microchip developed by the University of Twente. By adding blood plasma from patients to this model, Van der Meer and his team hope to develop models of COVID-19 patients. These models can then be used to simulate the development of blood clots. To realise this, they sought collaboration with Saskia Middeldorp, Professor of Internal Medicine at Amsterdam UMC, and Christine Mummery, Professor of Developmental Biology at Leiden University Medical Center. For the research, it is vital that material from different patients with and without COVID-19 is used. Because why do some COVID-19 patients suffer from these clots and hypoxia, whereas others do not? These individual models will function as mini-patients on which treatments and drugs can be tested in the second phase. For Van der Meer, the use of these human models is a logical step: ‘The technology has developed in such a way over the past 5 to 10 years that we can now make models that closely resemble humans and consequently make a fast “turn-over’ towards patients. The reflex of using animal models in medical research is deeply rooted. However, with the extra tools that are now at our disposal, we can do exciting things that have added value and are animal-free.’
Patients recover slowly from COVID-19, and it seems that both the virus and the immune system’s response to the virus cause damage to the alveoli. How do the cells that cover the respiratory passages and alveoli, the epithelial cells, respond to the virus and how does that response contribute to lung damage? These are the questions that Pieter Hiemstra, Professor of Cell Biology and
Immunology of Lung Diseases, will tackle together with his colleagues from Leiden University Medical Center. For the research, the team will make use of tested human cell models, organoids andconventional culture models. In the first phase, epithelial cells from, among other things, the nose and alveoli will be cultured to examine what the virus does with the different cell types.
Subsequently, the researchers will examine the response of the cells to infection with different coronaviruses, including SARS-CoV-2 that causes COVID-19. With this, Hiemstra hopes to discover what makes the COVID-19-virus so unique. The third step is to make a comparison between models
with cells from COVID-19 patients and healthy people. With that knowledge, the researchers expect to gain a better understanding of the behaviour of epithelial cells and immune cells in the case of COVID-19, to improve human models and to use these for testing drugs, for example. In the fourth phase of the project, the research group wants to develop a fibrosis-lung-on-chip. That can be used to investigate why the virus causes so much damage to the lungs in the short and long term. The ultimate goal is to improve the treatment of patients.
Hiemstra sees many advantages in using these human models. These make it possible to investigate differences between individuals, which is not possible, for example, with homogenous mouse models. Furthermore, with the human cell models research can be done on a large scale which leads to better results. Hiemstra: ‘With the funding from ZonMw and the Dutch Society for the
Replacement of Animal Testing, we can improve and refine the human models so that we can increasingly approach the situation in humans. The validation of animal-free innovations is a vital step in improving medical research and preventing pointless animal experiments.’
Dick Tommel, chair of the MKMD committee: ‘As a committee, we hope that with these projects, we can demonstrate that animal-free innovations can become the standard and animal research will become less of a norm. This is partly because no good animal models are available for research into the virus and COVID-19.’
The Dutch Society for the Replacement of Animal Testing.
The Dutch Society for the Replacement of Animal Testing seeks to realise a world without laboratory animals. One of the most important ways to achieve that goal is funding animal-free research. To this end, the Fund Animal-Free research was established in 2009 in collaboration with ZonMw. Since the inception of this fund, the foundation has funded various animal-free research projects that are not only better for the laboratory animals but also for people. The ultimate aim is a science based on the human situation, with innovative techniques that make the use of laboratory animals redundant.
ZonMw – More Knowledge with Fewer Animals
For more than 20 years, ZonMw has invested in research into the reduction, refinement and replacement of animal experiments. With the programme More Knowledge with Fewer Animals (Dutch acronym MKMD) ZonMw encourages innovations that contribute to answering scientific questions without animal experiments. The aim of the programme is to develop new animal-free innovations and to encourage the use of existing animal-free innovations. The ultimate aim is more relevant and improved health(care) research for people. ZonMw realises MKMD on behalf of the Ministry of Agriculture, Nature and Food Quality (LNV) and in doing this, it works together with various parties including the Dutch Society for the Replacement of Animal Testing, Top Sector Life Sciences and Health (LSH), civil society partners such as health foundations and the umbrella organisation Association of Dutch Health Foundations (SGF).