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Research

 

In the lab, fundamental diabetes research


 

By investing in fundamental scientific research, we hope that one day diabetes will be a thing of the past. The research projects we support are carried out by top researchers in renowned laboratories worldwide. Currently, the DON Foundation finances up to five research projects. These are carried out in the following laboratories: Hubrecht Institute, Leiden University Medical Centre, Vrije Universiteit Brussel, Hebrew University of Jerusalem and Expertisecentrum Bètacel Bescherming (Expertise Centre Beta Cell Protection). Unlike other organizations, an important part of our strategy is to commit to long-term agreements to give researchers sufficient time to fundamentally test their hypotheses.

 

Expertisecentrum Bètacel Bescherming (Expertise Centre Beta Cell Protection)

 

 Correcting a deficiency in the immune system


 

Type 1 diabetes is caused by a deficiency of the immune system. This deficiency causes the immune system to destroy the beta cells. Without the beta cells, less insulin is produced and, consequently,patients with diabetes need to inject insulin. Bart Roep examines how this immunological deficiency leads to diabetes and how this can be prevented or remedied. “What we really want to know”, says Roep, “is to which signals the beta cells of the immune system actually respond. Apparently, in some cases, the beta cells are telling the immune system: destroy us! At the same time, I am looking for ways to silence this request. If the immune system is able to ignore the beta cells, then the destruction of the beta cells, and with it the process of the disease that leads to diabetes, can be stopped. This is the most important step towards a cure.”

Stressed cells

De groep van Roe In the first year of his research supported by DON, Roep has achieved significant results. Beta cells assemble in the islets of Langerhans in the pancreas. If the islets becomestressed, then changes sometimes occur in certain proteins on the outside of the beta cells. Roep: “This creates ‘foreign’ proteins, which tells the immune system that the cells must be eliminated. Everything that is foreign in the body – such as viruses, bacteria and cancer cells – in principle are attacked and destroyed by the immune system. In diabetes, the immune system accidentally attacks healthy but stressed tissue.” The enzyme that sets the change of proteins in beta cells into motion is the same enzyme that converts gluten into an allergen in patients with celiac disease (a disease that occurs relatively frequently in patients with diabetes). Roep: “We are finally starting to understand which factors play a role in the development of diabetes. And so we finally have the opportunity to intervene in this process and to prevent the disease in people at high risk.”

 

First vaccine against diabetes

For the treatment, and perhaps even a cure, Roep’s research has reached a significant and exciting phase. “We have developed a cellular therapy,” says Roep, “which can prevent and limit destruction of beta cells. We will now start testing this cellular therapy in humans. As is usual, we must first determine whether this therapy is feasible and safe in humans. We expect it is, as we use the patients’ immune cells for the treatment.” How does it work? Roep: “After taking blood samples, we use vitamin D3 among other things to change the cells, and we cultivate them further with a vaccine that we have developed from pro-insulin. When we inject these treated cells back into the patient’s bloodstream, the immune system will actually learn to protect the islets of Langerhans. We have now been given permission to test this advanced cellular therapy; the first vaccine that has been developed in the Netherlands to stop diabetes!”

Customised therapy

Recently, we gained an important new insight, namely that there is such a large variation between diabetes patients. In other words: one case of type 1 diabetes is not the same as another. Roep: “This partly explains why we have not been able to find a ‘magic bullet’, and why we still do not have a therapy that is effective for everyone. In the coming period, we want to chart the differences in the disease among patients, in order to be able to offer each individual patient the most suitable therapy.”

 

It’s a lot of hard work, but ultimately we are convinced that diabetes can be eradicated. One way or another.

 

Prof. dr. Bart Roep Ph.D.

Expertise Centre Beta Cell Protection

Prof. dr. Bart Roep Ph.D.

  • Immunologist, diabetes researcher at the LUMC
  • Received 1.5 million EUR for research into a vaccine against type 1 diabetes, jointly from the DON Foundation and the Diabetes Foundation.
  • Notable quote: “People are sceptical: ‘So much money for research that is taking so long? Surely, it’s not rocket science? No, it’s harder than rocket science! NASA receives 20 billion per year, but Man has still not set a foot on Mars. We do our research with a lot less money, and it is at least as difficult!”

 

Hubrecht Institute

Cultivating detected stem cells in 3D


 

An important aim of the Hubrecht Institute, which specializes in developmental biology and stem cell research, is to identify, isolate and characterize stem cells that are located in the pancreas. Why is this so important? In type 1 diabetes, the islets of Langerhans in the pancreas are destroyed by its own immune system. Also, the beta cells found in the islets, die in the process. Only substitution of beta cells can cure the disease. Because there is a huge lack of donors, the solution appears to have to come from stem cells. Stem cells are cells that can make an infinite number of new cells. This is why stem cells in the pancreas are so interesting. They are an inexhaustible source of new cells which, moreover, are already on their way to becoming insulin-producing cells.

The pancreas

Against this background, the research sponsored by DON began in 2010. Initially, the study used the three-dimensional (3D) culture system for stem cells, developed by the group led by Hans Clevers. In 2013, it was demonstrated that pancreatic cells from mice could be cultivated in a tray, and that they mature into insulin-producing cells. However, mouse cells are not suitable for humans. In collaboration with the Leiden University Medical Centre (LUMC), cultivation and growth of human pancreatic cells was started. Insulin-producing cells also needed to be formed in the resulting clusters of cells (organoids). Eelco de Koning, who is linked both to the LUMC and the Hubrecht Institute, explains: “In 2014, we further optimized this culture system. We can now identify and isolate a specific population of these cells growing in a nursing tray and generate new cell clusters from a single cell (a progenitor or stem cell). With this, we have demonstrated that there are populations of cells with stem cell characteristics in the pancreas, and that we are able to make these cells divide over a longer period of time.”

Discoveries

In 2014, research was completed on stem cells in the intestinal wall that are able to create hormonal cells and are therefore important in the optimization of the islet function. A variety of signals has been discovered, enabling us to influence the formation of hormonal cells from stem cells in a 3D culture environment in the laboratory. Alexander van Oudenaarden – the new director of the Hubrecht Institute – and his group, analysed which genes can be read from tissue samples in individual cells. A huge leap forward, according to De Koning: “This gives us information about the function of individual cells, something that, until now, was impossible. We can use this technique to identify cells that have the characteristics of a stem cell. With the right ‘molecular bait’, we can very accurately catch the stem cells from the pancreatic tissue.”

Biotechnological platform

For the coming period, this wealth of information will be used to learn more about the characteristics and function of stem cells in the pancreas. The team wants to identify surface proteins, in order to use them to isolate these cells. Furthermore, we need to find the signals that induce these cells to grow and mature in the 3D culture systems. De Koning: “This is how we can create a biotechnological platform that is not only important for new cell therapies, but that may also be of major importance in studying regeneration of the pancreas in a laboratory. All this with the aim to encourage the recovery of damaged cells in patients with diabetes.”

In 2014 my research team won the ‘Breakthrough Prize’. This was because we discovered how to detect pancreatic stem cells, remove them in their purest form, and transform them in a laboratory into very large amounts of tissue. This was a global breakthrough.

Our discovery has brought us one step closer to creating a correctly functioning pancreas.

prof. dr. Hans Clevers Ph.D.

Hubrecht Institute

Hans Clevers - stichting DON

Prof. dr. Hans Clevers Ph.D.

  • Geneticist, diabetes researcher at the Hubrecht Institute, Professor at the University Medical Centre [UMC] Utrecht
  • Received an amount of 750,000 EUR from the DON Foundation for research into beta cells
  • Notable quote: “One way or another, it is very glamorous to commit yourself to the cancer cause. We will all be climbing the Alpe d’Huez, and we’ll be growing a moustache in November. All very well, but diabetes certainly deserves as much attention.”

Vrije Universiteit Brussel

From mouse to man


 

The research group of Harry Heimberg focuses specifically on how the remaining beta cells in diabetes patients can be protected and in what way new beta cells can be created. This group was the first to demonstrate the existence of the beta cell precursors; these are progenitor cells in the pancreas which are a source of new pancreatic cells. Together with Patrick Collombat (Université de Nice), Heimberg showed that alpha cells can be transformed into insulin-producing cells. Like beta cells, the alpha cells are located in the islets of Langerhans, but they secrete the hormone glucagon. These alpha cells can be transformed into beta cells that secrete insulin. Heimberg: “In this study, we used mice exclusively. We expect that in the near future we will be able to show similar findings in humans and thus contribute to a cure for diabetes.”

Two ways to cure diabetic mice

In recent years Heimberg succeeded in curing diabetic mice in two different ways. Heimberg: “In the first case, we used a technique in which we were able to reprogramme the pancreatic cells. With this method we were able to change cells, that under normal conditions produce digestive enzymes (acinar cells), into beta cells. Heimberg: “We demonstrated that we could transform alpha cells into correctly functioning beta cells by activating one very specific protein in the alpha cells. Furthermore, they proved, together with researchers from the Hubrecht Institute, that duct cells from the digestive enzymes can be infinitely multiplied in the pancreas and transformed into beta cells.

Transposing to humans

Heimberg and his colleagues focused on transposing their research from mice to applications for humans. They were successful in transforming the acinar cells, as well as the duct cells from the pancreas of humans, into beta cells. Heimberg: “After transplantation into diabetic mice, we saw that their sugar levels were restored. This offers prospects for the future, because there are approximately one hundred times more acinar cells and duct cells in the pancreas than beta cells. Add to that the fact that you can multiply duct cells infinitely, so there is hope that we can tap into an unlimited source for the production of beta cells in the patients themselves. “Slowly but surely, Heimberg can see excellent prospects for clinical research in humans. “It is possible to create beta cells from various different cell types that occur naturally in the pancreas, in abundance,” he says. “We can safely transform these cells into beta cells in the laboratory and then cultivate them in huge numbers for use in humans. But the transition from lab to clinic is complex and expensive, and with every step safety must be a priority.

Recent achievements

In the meantime, the research continues. Heimberg outlines some recent achievements: “Alpha cells are also replenished and regenerated. The alpha cells are essential for optimal functioning of the beta cells. We have found that the multiplication of alpha cells is dependent on interleukin 6 (IL-6). Furthermore, we saw that for a spectacular doubling of the beta cell mass, a number of growth factors are required. One of these is the well-known hormone oestrogen. And finally, important to mention: immune cells seem to play a part in the regeneration process of pancreas cells. That too has our attention.”

Prof. dr. Harry Heimberg Ph.D.

  • Researcher at the Diabetes Research Centre in Brussels and professor at the Vrije Universiteit Brussel
  • Is funded by the DON Foundation since 2012: for five years he will receive an annual amount of 100,000 EUR for research
  • Together with his research team has managed to cure diabetic mice.

Hebrew University of Jerusalem

 The role of cellular ageing in diabetes patients


 

Loss of insulin-producing beta cells and a lack of production of the same cells increases the risk of diabetes. The fact that beta cells find it hard to multiply is now common knowledge, and that the ability to do so further decreases with age. Yuval Dor: “We are interested in the molecular mechanisms that prompt this process. Which biological processes dictate the decrease and cause beta cells to release less insulin with increasing age?”

Switching the genes on and off

For this study, Dor’s group genetically modified a number of strains of mice. “We did it in such a way,” says Dor, “that we were able to concentrate specifically on switching the genes that regulate normal ageing of cells in these mice, on and off. This gives us the chance to speed up or slow down the ageing of beta cells and to see the effect it has on how beta cells function. It enabled us to determine the effects on the cellular programme that regulates the beta cell division, and also what effect beta cell ageing has on insulin secretion. We then investigated whether these results are also valid in humans and what consequences this would have for developing diabetes.”

Programme of cellular ageing

In the course of 2014, this research led to important new results, which are expected to have a considerable impact in the research field. The first results are to be published soon. Dor: “By and large, it includes the following. We have found that in young adults, genes that prevent beta cells from dividing, become active. This means that from this age on, the probability of regeneration of the pancreas and more particularly of the beta cells, is significantly reduced.” Surprisingly, they found that the same genes improved the function of the beta cell, as from that moment on, the cells are able to secrete more insulin. “What we can see is that, in fact, a complete programme of cellular ageing – cell senescence – is launched in the adult beta cells,” says Dor. “This programme not only affects the size of the cell, but also the metabolic rate, and many other cellular processes. We might also use this cellular programme to improve glucose control in diabetic mice.”

Challenging insights

It had not previously been demonstrated that cellular ageing affects the function of the beta cell to such a profound extent. These intriguing results now form the basis for new, challenging insights into the role of cellular ageing and cellular development; certainly in relation to diabetes. Dor: “Due to the discovery of this programme, we now have more understanding of why beta cells sometimes function better. Until now, we were not always able to figure out what caused these issues. Knowledge from this cellular programme also provides excellent opportunities for the development of new treatments for diabetes and applications in regenerative medicine.” Dor and his research team intend to publish the results of this research in a leading journal in 2015. “We also want to better understand how this ‘ageing programme’ effects the tolerability of beta cells when damaged and stressed, which can lead to diabetes,” says Dor. “Moreover, we are extremely curious to know whether this cellular programme also changes the interaction with immune cells. And if so, what does the change look like? This can provide useful knowledge for the development of an effective therapy against diabetes.”

As we age, the division of beta cells decreases, and their efficacy is reduced. Through genetic manipulation in mice, we have managed to create a model in which a gene is activated to combat the ageing of beta cells. Surprisingly, we discovered that in response to glucose, these cells secrete more insulin than very young beta cells. Older cells may be much more effective in the event of transplants than young beta cells. These are startling results!

Professor Yuval Dor

Hebrew University of Jerusalem

Prof. Yuval Dor

  • Professor and head of research at the Hebrew University-Hadassah Medical School in Jerusalem
  • Is funded by the DON Foundation since 2012: for five years he will receive an annual amount of 100,000 EUR for research
  • Together with his team, he is researching how insulin-producing cells grow, multiply and disappear in the pancreas

Leiden University Medical Centre (LUMC)

Natural mass production of beta cells


 

Eelco de Koning is searching in mice and humans for new treatments to replace destroyed pancreatic and beta cells in diabetic patients. The LUMC is already transplanting a complete pancreas or islets of Langerhans in patients. Both contain beta cells that produce insulin. Due to a lack of donors, these transplants, which can put an end to diabetes, are carried out too seldom. For this reason, De Koning is looking for new treatments for large groups of people.

New research opportunities

In 2014, De Koning and colleagues performed a new form of islet transplantation on people with a chronic inflammation of the pancreas. If the entire pancreas is removed in these patients, a very serious and difficult to treat form of diabetes arises. De Koning: “We can remove the islets of Langerhans from this kind of inflamed pancreas, and return them to the patient a few hours later, after surgery, via an infusion in the liver. The patients can then continue their lives with little or no insulin.” This ‘auto transplantation’ of islets of Langerhans offers new research opportunities. Medication against rejection of the islets is not necessary, because the transplanted tissue is the patient’s own tissue. Basically, you can treat this group of patients with their own stem cells from their own pancreas, which is able to produce large amounts of pancreatic tissue and therefore also beta cells. Create sufficient numbers of these cells and then give them back again. In the near future, we hope to test this cell replacement therapy in this patient group.

Metamorphosis

In 2014, research was strongly focused on regeneration of the pancreas. De Koning: “Once we understand everything about the development of the pancreas with the islets of Langerhans, then we can use that information to treat the pancreatic tissue in the laboratory in such a way that a huge amount of insulin-producing cells can be produced. Together with the Hubrecht Institute, we can now grow foetal tissue in a nursery tray. This is a very important step, which shows that we can mimic the development of the pancreas. We can transplant this foetal pancreatic tissue in mice and monitor its growth. Now we want to detect the factors that are crucial in the development of the pancreas and islets of Langerhans. De Koning is also examining mature beta cells as a source for creating insulin-producing cells. Normally, adult beta cells do not divide, however, they do if they first take on a different guise. De Koning: “That metamorphosis can be seen as a step back in time, whereby the cell regains new development opportunities. The underlying idea is that this cell type – the beta cell in a different guise – has a better chance of multiplying and developing into insulin-producing cells. Knowledge of this process is hugely important for the new generation of cell therapies.”

Biological Innovation

The search for new sources of insulin-producing cells, retaining the pancreas identity and protecting these cells from the immune system, are the mainstays of De Koning’s study. “We focus mainly on biological innovation. This means we try to create new beta cells from stem cells, in which case the beta cells must continue to function correctly. In the development of bio materials, we are collaborating with the University of Utrecht. Ultimately, our goal is that the Leiden Diabetes Centre develops new cell therapies that can actually help people with type 1 diabetes.”

My team has successfully managed to grow beta cells, but the cells are still immature. Cultivating these cells is one thing, but stopping the growth to prevent them from forming tumours, is another matter. As a result, we have not yet been successful in cultivating sufficient cells that do what we want them to do: provide the correct amounts of insulin at a given glucose value. In short, we are on the right path but we are not there yet.

prof. dr. Eelco de Koning Ph.D.

LUMC

Prof. dr. Eelco de Koning Ph.D.

  • Internist and endocrinologist, diabetes researcher at the LUMC and Hubrecht Institute
  • Received an amount of 750,000 EUR from the DON Foundation for his research into beta cells
  • Notable quote: “Of all the non-terminal chronic diseases, diabetes is the disease with which the patient is confronted with treatment most frequently. It has a huge impact on patients, and on their environment.”