PROVIDENCE – The story has many beginnings. Dr. Leslie De Groot, raised on a dairy farm in upstate New York, where he began milking cows when he was five years old, attended Union College and graduated from Columbia Medical School, to become one of the world leaders in endocrinology research.

During his career spanning six decades, much of his laboratory research focused on the autoimmune thyroid disorder known as Graves’ disease.

His daughter, Dr. Annie De Groot, who co-founded EpiVax in 1998, developed pioneering computer tools known as immuno-informatics. The work has focused on identifying the properties of infectious and autoimmune diseases and the links to Tregitopes [discovered by EpiVax], an epitope that can turn off an immune response.

Instead of working in vitro, lab studies done in a controlled environment outside of a living organism, or in vivo, using a whole, living organism, much of the research work done by EpiVax originates in silico, research and analysis performed on a computer.

Dr. Leslie De Groot admitted that he was not initially an early proponent of immuno-informatics, as he explained to ConvergenceRI in a recent interview.

“My training was at the experimental level, doing research, and which meant drawing blood and injecting animals, looking at the thyroid,” he said. “The idea that you could gain the same information by using a computer was little surprising and off-putting.”

Although he was one of the first employees at his daughter's Institute for immunology and Informatics at the University of Rhode Island [iCubed], moving his laboratory there in 2009, his work and his daughter’s work began from opposite ends of the spectrum.

“Annie was working on developing immunity to prevent a disease, and we were working on preventing immunity to prevent a disease,” Leslie De Groot said.

We did the same tests, he continued, but we were looking at the results from opposite directions.

“That’s exactly right,” Annie De Groot concurred. “We were trying to vaccinate, and he was trying to tolerate.”

What changed the relationship was the discovery of Tregitopes – an epitope that turned off immune responses. “It became evident that it could apply to autoimmune diseases,” Annie De Groot said.

A second beginning
Some 10 years ago, in 2008, the two Dr. De Groots applied for a joint SBIR award to use the novel Tregitope approach to develop a new way to treat Graves’ disease. It was envisioned as a collaborative enterprise by the De Groot father-and-daughter team. However, the research grant proposal was rejected.

Fast forward to 2017, when Eduardo Guillén, scientific director of protein therapeutics at EpiVax, starting from scratch, with a blank slate, as he described it, developed the idea for a research grant proposal focused on using the Tregitope approach to treat Graves’ disease, unaware of the previous grant application.

When Guillén approached Annie De Groot with the idea, she was delighted, and promptly shared all the previous research conducted to support the 2008 grant application.

And, on Sept. 1, 2017, SBIR awarded EpiVax a two-year, $177,020 grant for research entitled, “Novel Antigen Specific Immunomodulatory Tregitope-based Therapy To Address Autoimmune Pathogenesis in Graves’ Disease.”

While Leslie De Groot, at 89, will not play an active role in the research, his decades of research on Graves’ disease will certainly inform the effort that will be led by Guillén and Sandra Lelias, project manager of the Tregitope group at EpiVax.

[When asked about whether he would participate in any of the research, Leslie De Groot’s answer was an emphatic, “No,” adding: “I conducted hands-on research from 1956 to 2014. That’s a long time. I would spill the test tubes at this point.”]

“We have a great team,” Annie De Groot said. “To me, all of the stars finally aligned for Tregitopes and Graves’ disease, even though it took 10 years to get funded, we now have the opportunity to carry my dad’s work forward.”

Her father responded: “That would be quite wonderful.”

He continued: “If one could inhibit an immune response easily, and permanently, that would be quite an achievement. Suppose the same effect could be achieved with Type 1 diabetes. That would be mind blowing.”

Here is the ConvergenceRI interview with Dr. Leslie De Groot, his daughter Dr. Annie De Groot, Eduardo Guillén and Sandra Lelias of EpiVax, as the scientific research baton is passed from father to daughter.

ConvergenceRI: Can you talk about what generated your original interest in Graves’ disease?
LESLIE DE GROOT: I was a practicing endocrinologist interested in thyroid disease, in particular Graves’ disease and other autoimmune thyroid diseases. I spent my entire clinical life taking of care of patients with Graves’ disease, or Hashimoto’s thyroiditis, which is closely related.

ConvergenceRI: As Annie described it to me, at a family gathering, when you and she huddled together to talk science, she once bet you that her immuno-informatics tools could identify the peptides involved in Graves’ disease. Is that correct?
LESLIE DE GROOT: Yes. I was working on the causes of autoimmunity, and she was working on how to prevent autoimmunity.

The techniques were essentially the same. Basically, I moved into her lab [at Brown University, when she was a professor there], because she had a lot of expertise in immunology. So, it was very logical to work together. I believe that was back in 2004.

ConvergenceRI: Annie also mentioned that you went back for training in molecular biology.
LESLIE DE GROOT: In about 1990, I realized that the world was moving in a direction that I wasn’t trained in. So I went back and spent a year at Mass General Hospital where I trained as a resident.

It was in the pre-molecular biology era, although it was starting then.

ConvergenceRI: What is it like to collaborate as a father-daughter team in research?
LESLIE DE GROOT: I have great admiration for Annie. She has really developed quite a program and I am very proud of her work.

I have learned a lot from the team at EpiVax. If someone has something to teach, I’m ready to listen.

ConvergenceRI: How did the new successful SBIR grant evolve from the original 2008 grant proposal that was rejected?
GUILLÉN: I was dimly aware of the previous grant applications, but I started from zero, with a blank slate. I knew that I wanted to apply for a grant on autoimmune disease. I asked myself: what’s one of the most prevalent autoimmune diseases? Graves’ disease.

We have patients suffering from this disease, so we can get access to the patients in order to get samples from them.

And, with the tools that we developed at EpiVax, we can ameliorate the disease. I don’t like to say: “curing,” because curing is a big word. Rather, to focus on ameliorating the disease, improving the conditions of the patient, and helping the patient.

ConvergenceRI: How many people are affected by Graves’ disease?
LESLIE DE GROOT: I think it’s about 3-5 per thousand.

But Graves’ disease is one manifestation of thyroid autoimmunity disorders.

It commonly manifests itself as thyroid enlargement, and it affects some 20-50 percent of all women; it’s extremely common.

Graves’ disease is at the other end of the spectrum, but the autoimmunity is very similar.

ConvergenceRI: Continuing with the story of the grant, what happened next?
GUILLÉN: I mentioned to Annie that I wanted to write a grant on Graves’ disease, only to discover that we had all this work and all these papers and studies which applied directly to our grant.

Many of the research findings in the papers of which Dr. De Groot is an author were directly incorporated into the grant.

LESLIE DE GROOT: We worked on Graves’ disease for years before one could actually understand much of immunology. Immunology is a science that’s grown up in the last 30 or 40 years. I began working on Graves’ disease well before that.

ConvergenceRI: How many years ago did you begin working on it?
LESLIE DE GROOT: I think first papers on thyroid autoimmunity were published in about 1960, which would be almost 60 years ago. I was working at Mass General as a fellow.

The story of Graves’ disease as an autoimmune disease has been developing over the time that I’ve been in medicine.

I can remember when the first paper on autoimmune antibodies in thyroid diseases was presented at a conference in a tiny room in New York City at an Endocrine Society Meeting, in about 1958.

People didn’t know anything about autoimmunity and didn’t know anything about the cause of Graves’ disease; it was thought that it was possibly due to some kind of abnormal thyroid simulating hormone from the pituitary gland.

Gradually, it was recognized that the cause was related to autoimmunity in terms of antibodies and the thyroid, and then finally, that T-cells were involved, and that T-cell immunity was probably more important in thyroid autoimmunity.

The whole science of immunology developed in the period from 1960 until now.

ConvergenceRI: How have the studies of Graves’ disease evolved?
LESLIE DE GROOT: At first, we studied how to treat thyroid disease by giving people radioactive iodine and destroying the thyroid.

Then we started to study autoimmunity, trying to find a way at first to understand what autoimmunity was, and what aspects of the thyroid and what proteins are involved in autoimmunity.

Then there was this amazing discovery by a doctor in New Zealand who found that thyroid blood, or serum, really, from patients with thyroid disease stimulated the thyroid of animals.

It turned out that the stimulating factor was an antibody.

What’s going on in this grant is an attempt to alter the immunology, if not suppressing it, or least turn it down, by using an approach Annie and her group have pioneered. Let’s hope it works.

ConvergenceRI: Are you going to be working with mouse models?
GUILLÉN: We are first going to be working on selecting peptides in lymphocytes from patients suffering from Graves’ disease, in order to target the Tregitopes to the right cells, which are the cells that are reacting to the TSH receptor.

Which peptides to select have largely been studied by Dr. De Groot. We are going to test those peptides in the blood from Graves’ patients, to confirm that they have stimulatory effects.

And then we are going mix those peptides with Tregitopes to show that the stimulation doesn’t happen anymore.

Then, we plan to select the peptides with Tregitopes that work and test them, using a mouse model of Graves’ disease.

LESLIE DE GROOT: The immunity that causes Graves’ disease is linked to a protein on the surface of the thyroid, called the TSH receptor. Everything in disease works with a messenger and a receptor.

And the messenger is the pituitary hormone, TSH, and the receptor is a protein that messenger binds to and starts signaling.

That’s the way everything works; I guess it works that way with people, too.

ConvergenceRI: I have to take some time to think about that one. Can you talk about the nature of collaboration in research? Particularly when you started at different ends of the spectrum, with different ways of looking at the world. Annie described you as a PC person and herself as a MAC person.
LESLIE DE GROOT: I guess that’s true.

ConvergenceRI: What have you learned from each other?
LESLIE DE GROOT: I’m not exactly sure how to answer that question.

My training was at the experimental level, doing research, and which meant drawing blood and injecting animals, looking at the thyroid. The idea that you could gain the same information by using a computer was little surprising and off-putting.

I think there are two parts that made for success in the informatics approach. The first part is that informatics were built upon laboratory experiments.

The idea of what’s an epitope and what cells are involved came from hard research on animals, and then it was applied to look at immune phenomena in silico.

I think that while the laboratory approach may still have a very crucial role, because of the advances of immunology, it is possible to duplicate some of the in vitro research in silico.