A great deal of knowledge about the SARS-CoV-2 virus and the COVID-19 disease is needed quickly. In April this year, the Dutch Ministry of Health, Welfare and Sport (VWS) made 40 million euros available to ZonMw and NWO for COVID-19. Experimental animal-free innovations enable research to be carried out better and faster because the results can be translated more readily to humans. The committee of the ZonMw research programme ‘More Knowledge with Fewer Animals’ therefore made extra money available to enable research with such non-animal testing innovations within the COVID-19 research programme and selected five animal-free science projects that will enable wider use of existing non-animal innovations or the development of new non-animal innovations.
A puff of heparin against corona infection? – Theo Geijtenbeek (Amsterdam UMC)
Professor Theo Geijtenbeek, Professor of Molecular and Cellular Immunology at the Amsterdam UMC, will lead the research into the possible preventive effect of the anticoagulant low molecular weight heparin against SARS-CoV-2. COVID-19 patients are already receiving heparin immediately upon admission to hospital by means of injections to prevent blood clots. However, Geijtenbeek and his group discovered that this drug also blocks the virus’ binding to cells and therefore prevents infection. They now want to investigate whether the inhalation of heparin can have a preventative effect so that healthcare staff, can protect themselves against infection with heparin inhalation. The first step in the research is innovative, says a proud Geijtenbeek. Volunteers are asked to inhale heparin through the nose. The researchers then remove some cells from the nasal mucosa (as in a coronary test) and then expose those cells to the virus to investigate the antiviral effect of heparin. Geijtenbeek: “We want to do this in this way to avoid animal testing and get to the clinical phase sooner. And time is important in this pandemic”. In addition, the research group will use a dynamic human cell model to further investigate the effect of heparin on coronavirus infection.
A widely applicable dynamic cell model – Robbert Rottier (Sophia Children’s Hospital)
A dynamic cell model is a research design that the consortium led by Robbert Rottier, senior researcher at the lung department of the Sophia Children’s Hospital, is also working on. Current COVID-19 research makes extensive use of static lung systems. The disadvantage of these models is that they only imitate the functioning of lung cells in a human being to a limited extent. This is why the Rottier team, together with Professor Roman Truckenmüller of Maastricht University and the MERLN institute, will create a closed dynamic system. They will use an existing bioreactor to simultaneously grow human cells from both the epithelium of airways and blood vessels. By sending ‘microfluids’ through it, a dynamic system is created. In this way, the development and course of viral infections such as COVID-19 can be better studied. The collaboration with Truckenmüller, an expert in the field of nanotechnology and biochips, and the MERLN institute means that this system can also be quickly made commercially available to laboratories. The team is also working on a protocol so that the model can be applied without additional training. According to Rottier, working on animal-free innovations has additional advantages: “Since we have been working on this type of innovation, we are increasingly looking for alternatives within the research group and the use of laboratory animals has decreased.
Combining human cell models with genetic characteristics – Jeffrey Beekman (UMC Utrecht)
Working on non-animal innovations can make researchers more aware of how they conduct research. Professor Jeffrey Beekman, Professor of Cellular Disease Models at the UMC Utrecht, also experienced this: “Before, I was not so much involved in animal-free research, the call for funding and the writing of the application made me more aware of this. I am now even more aware of the materials I work with, such as the human cells that come from individuals and the serum that I use to grow the cells. They are often of animal origin. For COVID-19 research, Beekman and his research team will use cell models to study how in vivo the corona infection affects different organs: upper respiratory tract (nose), lower respiratory tract (lungs), intestines and kidneys. “By combining these models with the unique genetic characteristics of the donors of these cells, you can compare different tissues and identify factors that influence the effects of the coronavirus and the efficacy of medicines,” says Beekman, “which is particularly important in the case of COVID-19 because the virus affects different organs.
Microchips as mini-patient with COVID-19 – Andries van der Meer (University of Twente)
How can you mimic a COVID-19 patient so that you can investigate why some patients develop blood coagulation? That was the question with which Andries van der Meer, researcher Applied Stem Cell Technology at the University of Twente, and his team set to work. 10-30% – of the people who end up in hospital with COVID-19 develop blood coagulation. As a result, this group of people in particular has a much 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 patient blood plasma to this model, Van der Meer and his team hope to be able to develop models of COVID-19 patients. These models can be used to simulate the development of blood coagulation. To make this possible, they sought collaboration with Saskia Middeldorp, Professor of Internal Medicine at the Amsterdam UMC, and Christine Mummery, Professor of Developmental Biology at the LUMC. For the research it is essential to use material from different patients, with and without COVID-19. After all, why does one COVID-19 patient suffer from these clots and lack of oxygen and another does not? These individual models function as mini-patients on which treatments and medication can be tested in a second phase.
For Van der Meer, the use of these humane models is a logical step: “Over the past 5 to 10 years, the technology has developed in such a way that we can now make models that are close to the human being and can therefore make a quick ‘turn-over’ to patients. The reflex to use animal models in medical research is deeply rooted. But with the extra tools we now have, we can do exciting things that have added value and are animal-free”.
Better insight into lung damage by COVID-19 – Pieter Hiemstra (LUMC)
Patients are slowly recovering from COVID-19 and it seems that both the virus itself and the immune system’s response to the virus causes damage to the lung vesicles. How do the cells lining the airways and lung vesicles, the epithelial cells, react to the virus and how does that reaction contribute to lung damage? These are the questions Pieter Hiemstra, Professor of Cell Biology and Immunology of Pulmonary Diseases together with colleagues from LUMC, will be working on. For the research, the team will use tried and tested human cell models, organoids and conventional culture models. In the first phase, epithelial cells will be cultured from the nose and lung vesicles, among other things, to see what the virus does to the various cell types.
The researchers will then look at the cells’ response to infection with various coronaviruses, including SARS-CoV-2, which causes COVID-19. In this way, Hiemstra hopes to discover what is so unique about the COVID-19 virus. The third step is to make a comparison between models with cells from COVID-19 patients and healthy people. With this knowledge, the researchers expect to better understand the behaviour of epithelial cells and immune cells in COVID-19, to improve the human models and to apply them, for example, for drug testing. In the fourth phase of the project, the research group wants to develop a fibrosis-long-on-a-chip. This will make it possible to investigate why the virus causes so much damage in the lungs in the short and long term. The ultimate goal is to improve the treatment of patients. Hiemstra sees many advantages in the use of these human models. This makes it possible to investigate differences between individuals, which is not possible with homogenous mouse models, for example. With the human cell models it is also possible to do research on a larger scale which leads to better results. Hiemstra: “With the funding of ZonMw and Proefdiervrij we can improve and refine the humane models so that we approach the situation in humans more and more. Validation of non-animal innovations is an essential step in improving medical research and preventing useless animal testing”.
Non-animal innovations for better COVID-19 research
Dick Tommel, (Chairman of the FMD Committee): “As a committee, we hope that these projects will show that animal-free innovations can become the standard and that animal testing can become less and less. Partly because there are no good animal models for research into the virus and COVID-19”.