2018 Major Trends in Modern Cancer Research

Can I get everyone's attention? I want to welcome you all here. I'm Craig Thompson. I am the president of Immoral Son Kettering, so it is a real honor for us to welcome all the students here each year. This is our thirteenth year in the specialty.

trends

in

cancer

research

and we're always happy to have you here and hopefully get you a little excited about what could happen in scientific

research

. You've been exposed to it in your classes and now you'll get to see three of our best young scientists and these are some of the ideas they have about new approaches to

cancer

, so one of the things I want to do as we start is have a little discussion about why we really do research here for cancer.

Memorial Sloan Kettering is the oldest cancer hospital in the United States. We were founded right here, in this part of New York, one hundred and thirty-four years ago, when the germ theory of disease was just becoming fashionable, before the decade. 1880's when we started there was no appreciation that germs cause disease viruses bacteria the other pathogens we know now many of which you have had that made a home to you from school when you were younger we are not appreciated for what they were and then suddenly in the late eighteenth and late 1870s the germ theory of cancer was discovered.

We discovered that there were viruses. People like Pasteur discovered vaccines and approaches like that and from that intense interest in us being able to treat diseases came the discovery that there was a new disease that Scientists hadn't really understood that, in fact, it's not a new disease, it's been with us. for thousands of years, but it is cancer and it affects every organ in our body and it is a disease that is inexorable once cancer begins and a cell begins to divide. of control in the body, slowly but surely, first it takes over the tissue of its origin and then finally it learns to metastasize to live and, wow, in other parts of our body, and that is actually the devastating complication that we care for here at Memorial Sloan Hospital. -Kettering today, after 134 years, we can practically eliminate primary cancer from anyone with the great surgeons who are here.

We have one hundred and 25 surgeons who only deal with cancer, so they have studied for years to understand how they can safely remove it. a cancer in virtually every organ in your body, because every organ in your body is susceptible to cancer, there are cells as part of our ability to heal and regenerate that, as they proliferate to replace their tissues, can undergo mutations in their DNA, not the DNA is exactly what you inherited from mom and dad, but as they replicate the DNA that you got from mom and dad, they can make mistakes and it's those mistakes in the genes that regulate cell division and cell proliferation those that lead to cancer.

It's really been an exciting time in cancer biology over the last 20 years because we can now figure out what's wrong with each patient by sequencing the DNA of their cancer cells and distinguishing it from normal cells, and that's given us a tremendous amount of insight. tools. To begin to understand this disease of cancer now, how many actually know someone in their family or close relatives who has had cancer? Look around you, almost everyone is with their hands raised and that is because today we are living longer and we can to effectively treat other diseases such as heart disease, cancer will affect one in two of all men in the room, so all of us guys one and two are ahead, but it's not something we want to be ahead of.

I'm going to say that for all the women in the room the incidence is now over 40%. We used to say it was one in three, but women are catching up because, unfortunately, there are still things we do to ourselves that increase our cancer risk one by one. young women are smoking, the other is exposing ourselves to sunlight and going out in the summer and overcoming sun exposure and the third, actually, here in New York, what is a problem is the idea that obesity It is a tribute to cancer that we did not understand even ten times. years ago, so we're discovering new things and the type 2 diabetes epidemic that you may have learned about in your health classrooms is really increasing the risk of cancer as we go, so we're learning a lot about that, but now we'd like to finding ways to actually treat it and the last period of time here at Memorial sloan-kettering has been developing treatments to treat cancers locally as they occur or recur and that's surgery or radiation oncology techniques the problem. they only work locally they only work where you know the cancer is and today we know that cancer is the systemic disease once someone has a lump that is big enough in a tissue to feel it so you can feel it on a self exam or In fact, a doctor can feel it and see it when they examine you and the clinic, it's already in your circulation and spreading, so today, to have effective therapy, you have to treat patients systemically, you have to be able to understand how to receive treatments. that can affect every organ in your body and that leads to the side effects that people are afraid of the side effects of chemotherapy that you've heard about, like losing your hair from what was called cytotoxic chemotherapy, which were cells that they just damaged any cell that divides and that works for cancer that I had multiple cancers when I started cancer control because these chemotherapies would destroy every cell that divided.

The problem is that you have some cells in your body that divide all the time and that are healthy in every blood. cell that you have with you right now you have cut everything so you have seen your blood recently true every blood cell that you have now will disappear in a hundred days you will make them new again that is five liters of blood You are going to make something new so unfortunately we give it cytotoxic chemotherapy several years ago. What happened is you stopped producing blood cells and people became anemic and were at risk of infection, plus the whole lining of your skin, your hair is always growing. it's always replaced as you shed it while eating and your chest lining is also on your skin when you get cuts, bumps and bruises and your hair constantly grows, that's what you have to cut your hair periodically, all those weaves they stop. with cytotoxic chemotherapy that is why people lose their hair and you see people who have lost all their hair as part of the cancer treatment, those treatments were good because they showed us that we could cure people even when the Cancer and the spread of cancer today we recognize it unlike other things.

Like heart disease, which is a relatively limited number of diseases, there are 400 different diseases that make up cancer, think about each specialized tissue in your body, you are at risk of developing cancer and each tissue has its own memory of which tissue it comes from. and where it could go to colonize because that's what the cancer cell thinks it should do is start a new organ somewhere else and that's the real problem we're going to talk about today, understanding how we can take advantage of this information that cancer arises from. through mutations that you did not inherit and then understand this problem of metastasis, how two cancers, although they go into circulation, most of them do not survive, they do not know how to live elsewhere, but some do. and one of the most devastating places they can live is in your brain, so let's hear a little bit about the brain, how the brain becomes a seed and how we can treat that's a pretty interesting area right now.

I think a lot of people may I met Jimmy Carter a couple of years ago, a former president from long before you were born, but I'm still doing a number of really important world jobs. He became ill with a type of skin cancer called melanoma that metastasized to multiple places in his brain. and a treatment developed here by researchers who were in this building actually allowed him to be cured of that disease and that was by taking advantage of the immune response by taking advantage of his immune system to recognize the mutations that were in his cancer and recognize them just as you recognized a virus or a bacteria. and then you attacked them and destroyed them just like you did when you had a cold last time and it took you two weeks to get over it, but your immune system cured you of all that today.

We would like to extend that type of treatment to many artistic types of tumor cells, we can often do this by tricking our normal immune system, but one of the most interesting areas that can really lead to these cures is this phenomenon that you may have heard about in Their classes are called car T cells which means car sounds like a clever acronym it means chimeric antigen receptor every lymphocyte in your body B and T cells have a specific antigen receptor that they produce as they progress in development that gives them allows us to be ready to find a virus that is not you that can recognize and initiate an immune response.

We have learned how to clone these receptors with molecular biology techniques and now we can design antibodies that are against the tumor cell for the components that transmit those signals to the immune cell and then introduce them through gene therapy into our own normal immune cells and take advantage of their ability to generate an immune response against what we have taught them to recognize, which is hopefully a specific antigen, something foreign that has been created in your tumor cell today. It's been really exciting, the Car T cells that you've heard about have cured children of acute lymphocytic leukemia, which is the common form of childhood leukemia or have recently been shown to cure adults with leukemias and lymphomas, but those are the minority of cancers, etc.

One of the most interesting areas of research being carried out at this institution is trying to discover if we can take advantage of this technology that combines cells, are we hiding it?, that come from the cancer patient themselves, designing them to grow and produce millions of them . and then give them through gene therapy a new way to recognize tumors with specific aspects of the tumor and do it with solid tumors, so tonight to begin our presentations we have one of our surgeons, an expert in chest cancers. cavity, so that's lung cancer in the cells that line the lung cancer and he wasn't.

I would have to say that I was satisfied with just being a surgeon that could remove the primary tumor, many of those tumors spread to other parts of the chest and places that you just can't remove the lining of the heart and other places, so a few years ago years prasad isuzu Mele, who will be our first speaker, decided to read and teach himself molecular biology and cell biology and combine his extraordinary surgical skills. with his extraordinary skills in the laboratory and he will bring a new therapy to patients with lung cancer diseases and he will have the opportunity to tell you about it right now, so Prasad with that introduction, he will be in the outer rooms if you want. to ask questions in the room at the end of your talk, you will speak for about 20 minutes and then there will be Q&As that you can do directly through YouTube so that our YouTube social media team will answer the YouTube questions that will feed Enter the room here to do that, plus everyone in the room must be sitting here thinking about the questions that, OH, come to you that you didn't fully understand.

We want you to come away with a good appreciation of what's going on and so you should be thinking of a question, but there is one rule: ask your question. The first brave person will appear at the end of Prasad's talk. He has to get close to the microphone so that people in the outside rooms can hear you. award every year is just a recognition, you don't get money or anything nice, the best part of this for us is when one of you asks a question that stumps one of our speakers and he says: you know, I don't know, that's because what we're investigating and how we're going to find out, so you should pay attention to see if you can confuse the stars, so with that I'll hand the podium over to Prasad to tell you a little bit about car T cell therapy.

It was that good night, so thank you for the great introduction and I'm happy and excited to be here to present some of our lab research to all of you as dr. Thompson mentioned that what we do is immunotherapy for solid tumors, so cancers that arise from blood cells, leukemia and lymphoma, those are liquid tumors, lung cancer, breast cancer, brain cancer, these are all solid tumors and that's what I'm going to talk about. So as you heard, I am a doctor and surgeonthen we went back to the bench to study why we can't kill the whole cancer cell, of course we gave it a small dose.

Safety Study Card Working against solid tumors for card T cells is an arduous task because, as I said, solid tumors are very notorious for evading the immune response. I already mentioned that we made the car for a specific antigen. We use these reinforcements on the coast. Domain immolated due to advances in immunology, we know what genes to put in T cells to make them more powerful, but the harder we push the accelerator, the stronger the breakdowns occur, that is due to two reasons number one, mother nature puts brakes on the T cells. They, Mother Nature, doesn't want the T cells to stay on, she can damage the way the T cells are activated, the brakes are activated and cancer number two knows that this cancer learns this and the Cancer secretes proteins in the mechanism where they exhaust the copy. cell, so that's what we realized, although we are powering the car's T cells by giving it a lot of gas, but the brakes are revving, the cancer is pressing on these brakes, so, for example, that one is called PD, a PDL, an activated T cell expresses PD. programmed into the receptor one and the cancer cell is producing this PDL one, so when the PDL one binds to the PD, a car T cell is exhausted, it is there in the tumor where we can see it in the tumor when it is They take biopsies, but they don't do it. work, they are functionally exhausted knowing that we use this checkpoint blockade immunotherapy.

What I mentioned before, there are specific blocking antibodies that prevent the PDL one tumor cell from binding to the car T cell on the PD T cell here is an experiment where we know the mouse had a lot of mesothelioma cancer we give it a small dose of cells T you can see that the T cells kill the cancer the cancer went down the regression of the tumor volume but three weeks later the T cells of the car were exhausted and the tumor began to return so we give this antibody PD an antibody three doses now we can reactivate the auto T cells which are the living drug inside the patient Mouse tumor and the tumor volume regresses and we can continue to administer this antibody and we noticed that the mice live longer and we can regress the cancer, so when we came back to the clinical trial here There is a patient who is 73 years old, served on a battleship exposed to asbestos and contracted mesothelioma and this is all the cancer that can be seen around his right lung when the biopsy is done.

There is a lot of cancer here, we give a dose of auto T cells directly into the chest cavity using the scanners and then we give them a checkpoint blockade. You can see now that the cancer is completely removed after six months, when we did the biopsy there were only a few cancer cells left. Most of the cancer was gone and this patient did well for about 16 months, we thought any additional therapy was necessary. Remember the patient I mentioned where we gave a small dose of auto T cells directly, just a 40 percent reduction. We did the same thing we started managing. checkpoint blockade to reactivate car T cells to make them recover from functional exhaustion and so far we can't see any tumor and it's clinically fine how much time do we have to give him the antibody every three weeks we keep giving it that's the regimen to reactivate the car's T cells then I thought, why keep giving the antibody?

Let's use genetic engineering to find an alternative method. I told them this is the break that Mother Nature put in the PD T cell and the tumor cell kept pushing the break of the PDL, so we thought, what's going on? let's take out the transmission system inside, let's let the rest be there, let's take out the transmission system or the signaling cascade, so we eliminated the signaling cascade that J because of genetic engineering, we eliminated this now we made a decoy receptor, the automobile T cell junction chain. The agent is activated with the decoy receptor, the tumor cell will try to bind to it, but there is no signaling and the t'car T cell is no longer exhaustive and continues to kill the cancer cell and that is what we tell the mice, they They are mice with a high tumor burden and when we continue to give small doses of cod T cells we can eliminate the cancer and they can continue without exhaustion, but this is a delicate balance when using immunotherapy, we want the immune cells to be excited. system to fight cancer, but there is a delicate balance, so it does not work out in the city of Aqsa and that is where we do the clinical trials, so what I presented to you in the last 15 minutes is the last 10 years of work that have done the The whole group and the lab and several doctors here is where we are, this is what we are facing, but look at what is happening with cancer in the latest FDA approved cancer drugs that have taken off because of all advances in technology and science. and more and more patients are living after being treated for cancer and that is the excitement and the opportunity to advance science and engineering that you will be entering into in the next decade, so I would like to take a minute to thank my mentors who taught me. the lessons, great colleagues wherever you can do and all the research staff who did all this work, thank you, very good, questions for Prasad's talk, someone has to be brave, ladies in mind, someone is coming, yes, yes , especially when you created that. decoy receptor and you are allowing the T cells in the car to continue to be used.

I'm thinking how powerful these car T cells are. What is the potential incidence of a mutation in car T cells that creates a potentially super autoimmune disease? Yes, so you always have to equate safety with efficacy and that's what we do meticulously and systematically in the lab, not just in mice, we take samples from patients, the T cells have several other breaks, which we're exploiting is the The main one of the main ones breaks down the PDL pd-1, but the T cell has several other inhibitory receptors that can come into play. Number one. Number two. We know this can happen, which is why we monitor, for example, any gene therapy in the United States.

The FDA requires for 15 years that we have to follow this patient to make sure that there is no mutation as you mentioned and if it happens, if there is any damage, our hospital, that's where the clinical field comes in, is extremely, we have algorithms to treat the patient, what medicine to administer to reduce the toxicity, we can even eliminate the heart cell if we want, for example, in our clinical trial we put a safety gene that does nothing on a regular basis, but if something goes wrong, we can administer a medicine in half an hour, we can make it reach all the Karaka cells in the body, so the various steps of the clinical aspect that we present are safe.

Thank you. Other questions that were great. How is it explained? Cancer cells that are constantly mutating and changing, so are there new T cells that you would put back into the body for that? That is the ongoing investigation. I hope you come to the lab and do some of that research to find out that it does. You are absolutely right, cancer cells constantly learn to evade whatever we do, the immune attack, but that is where we believe that our natural cells, if we give it a drug, the drug takes care of one thing, it can be eliminated, but If we give it a cell, the cell will do it too.

It learns just like the cancer cell but in the opposite way and, more importantly, now that we have made a step, a car, a genetic engineering, we can introduce multiple genes or multiple drugs. We know that these courtesies go to the tumor. We can make them secrete a certain type of medicine. the agents, so this is an ongoing investigation, that's what when these patients receive the biopsies, we are very grateful for these patients who participate in the trial, we do the biopsies and we learn what stage we are in and if the cancer cell is waiting for what exact mechanisms what we are doing for T cells as well, as those T cells have specific things to attack a specific type of cell, as they can only attack that type of cell once the cancer cell mutates.

Could you do T cell therapy again to introduce a new type? of cell that would attack only that type of cancer cell, yes, what our grandmother told us, life is a balance, so we constantly balance your immune system, so cancer is doing this anti rotamer immune system, we are increasing it , we are maintaining balance by adding checkpoint blockade it is a constant balance and what we learn is that we do not have to go after every cancer cell if we tilt the immune system making one car one type of antigen two types of antigen once we manage to balance Can the body's immune system come into play and get rid of other cancer cells?

So this is a constant balance that we are learning and we are promoting anti-tumor immune responses. Exactly what prevents the body from rejecting those T cells. Is there a specific type of antigen that? you put them in to prevent the body from recognizing that the body didn't initially create the lessons that we learned initially because it's easy to take the mouse antibodies and do this course, that's what everyone has done, but the body is generating an immune response when I started , had the benefit of that knowledge, so, for example, in our car, for the first time in the world, we made completely human components, so technically, in theory, they should not be an immune response because the entire human component, but we have to wait and see, I think we made it.

To move forward now, so you should definitely connect and talk more later, so those were great questions because we're actually in the early days of harnessing those patient's own T cells that are modifying how. we do it effectively so that they don't cause other diseases, it's going to be an exciting time here in the research labs and as well as being one of a group of about 10 labs doing research on this, we're now going to move on to the other part of the metastasis problem, which is how it actually happens, and we're lucky to have Adrienne Hwa, who's here.

Adrienne is a triple threat, she trained as a neurologist and then as an oncologist, as well as brain cancers and she also earned a doctorate in basics. she science and she has combined all three to understand how cancer cells spread in the brain and who helps and who tries to prevent it, so Adrienne, you're awake. Thanks Craig, today I'd like to talk to you a little bit about metastases. to the brain um so what I'd like to start with is what we study, which of course is brain metastasis, how we study it, which of course is we start with what we know and we go from there and why we study it and we study it.

Ultimately, it is to help people and improve the lives of our patients, so our bodies are made up of highly specialized cells that are grouped together into organs in any of these cells, just as Dr. Thompson said that It can become malignant and when it does it grows uncontrollably and becomes a tumor and this is known as the primary tumor. Integral to this progression to becoming a tumor is acquiring the ability to live and grow elsewhere in the body. and this is known as metastasis, so cancer can certainly start in any of our tissues and can end up anywhere else, so metastasis is a defining characteristic of malignancy, in fact, it is one of the main criteria that They determine whether a tumor is benign or malignant and metastasis.

While we understand a lot about the details of how cells transform or gain the ability to grow uncontrollably, we understand very little about the process by which cancer cells can travel to other parts of the body or metastasize to one part of the body. body. The main conceptual frameworks for this come to us from the Victorian pathologist Stephen Padgett, so that's this guy, Stephen Padgett, who was looking at tissue samples and thought about the problem of metastasis and thought and wisely reasoned that if it were a process completely stochastic. If this were a process that was simply by chance, then an organ that received a lot of blood, like the spleen, should have a lot of tumors, true, it should have a lot of metastases, but that is exactly the opposite of what is happening and so he wisely made the observation that when a plant produces seeds, its seeds are carried in all directions, but they can only live and grow if they fall into pleasant soil and this is known as the seed and soil hypothesis and this conceptual framework colors this metaphor. the way most of us think about metastasis because it captures this really intimate relationship betweenthe cancer cell and its microenvironment and I would like you to see how this develops as I study metastases in the brain, so brain metastasis comes in various flavors the two main flavors are left open angie and parenchyma, so meningioma of Tacitus leptin is metastasis in the coverings of the brain which includes the cerebrospinal fluid and parenchymal metastasis which would be metastasis in the brain tissue, so unfortunately this site of metastasis is very common.

So at autopsy, the CNS metastasis rate reaches almost 25% and even more unfortunate is that very few people study central nervous system metastases, so my ulterior motive is to make you all very interested. in this so that one day they can Everyone will study brain metastases, so why don't people study brain metastases? It is a very complex problem. Any tumor can cause metastasis to the brain. The main culprits are lung cancer, small cancer, breast cancer, small cell lung cancer, breast cancer, melanoma, but. really anything can and as you know and judging from your questions, you know very well that these cancers will continue to mutate, they will continue to acquire new mutations and therefore the tumors themselves are evolving over time and in response to our treatments, And what happens if we apply that knowledge to the seed and soil hypothesis, what happens if we recognize and accept it?

What if we say you know? We are not really studying how a cell, a seed, lives in a single soil. What we are really studying is how various seeds live in the same soil, and perhaps by studying this process, by studying how various seeds live in the same soil, we will not only understand metastasis, but this will give us novel insights into how metastasis works. brain itself, so there are two main microenvironments that we have to deal with with brain metastases, one of them is the leptomeninges and the other is the parenchyma, so first let's dive into the leptomeninges, so the leptomeninges are the covers of the brain.

They are called leptomeninges because they are the soft meninges, these are the soft covers. It is a beautiful drawing by SE Read here where he shows us all the coverings of the brain and in particular these leptomeninges, so it consists of the pia mater that attaches directly to the brain and the arachnoid. and this contains the circulating cerebrospinal fluid that you can see here, so it's quite complex, it's a complex site of disease and it's a very unusual site of disease, if you look at that cerebrospinal fluid, it doesn't look like there's much of it. In it, we'll get to that later, but despite this, cancer still finds a way to grow in this space, so the big question I had when I started this project was really how can cancer live and grow in this? such a strange place.

To answer this complex question, I needed a model system that was complex enough to be able to model this properly, so we turned to the mouse, so in this system we used something called in vivo iterative selection, which is really a way of describe. That we take cancer cells from a diverse population of cancer cells and use a mouse to divide this population of cells so that we get a subpopulation of cells with the ability to live and grow in this other environment, I think that really is the best way to show Actually, this is just to show you a good example, one of the stasis leptomeninges models that I made first was of breast cancer, so you can see here, here is the head of a mouse that just had cancer cells, these parental cancer cells.

A heterogeneous population of cells is instilled directly into the cerebrospinal fluid and this rainbow color you are seeing is because these cancer cells have been genetically engineered to emit a photon of light. They carry the firefly luciferase gene, so this allows us to MANET non-invasively. take pictures of the mice and you can quantify how much cancer is in the brain of that mouse just by taking its photograph, so over time you can see that we start to lose signals, we are losing cancer cells because not all of these cancer cells have the ability to grow in this space, however, over time they take over this space, we collect those cells and inject them into the brain in the spinal fluid of the next mouse and over time we create a subpopulation of cells that has the ability to live. in the CSF we then take these cells and inject them into the heart of the mouse to spread them throughout the space and the cancer cells are really just the cancer cells that reach the leptomeninges and have the ability to grow within that space.

The only cells left are the leptomeninges tatak cells, so of course I didn't want to generate just a mouse model. I didn't want to generate just a seed to study. I needed a lot of seeds to study to really stack the platform, so we made four mouse models and armed with this. With these tools we were able to ask some really interesting questions, so the first question we asked, of course, was: do these models work? So you can see here. We observed these salobreña mice under a microscope and discovered that they do indeed work. They grow on the surface of the brain as expected and do not produce parenchymal metastases.

Some other models that I will tell you about later that do produce parenchymal metastases do not cause the leptomeninges to suffer stasis. Then we asked more or less. These are these subpopulations of cells, these lepto M cells, how are they able to grow in this space? What are they doing differently from the parental cells? And to answer that question, we took a look at their transcriptome, so we took a look at their messenger RNA and looked to see what was in common between all of these various cell lines. What did all these seeds that could grow in this place have in common?

We found 20 genes in common, two that we thought were really interesting. and one that I'm going to talk to you about today is called complement C 3 or just C 3 for short, so once we identified this gene, we asked ourselves if this important human disease was a series of fairly complex mouse models, where did we end up? ? We ended up in the right place and then I took a look at some of my patients' samples, so when looking at the cerebrospinal fluid of patients harboring leptomeninges and stasis we found that the cerebrospinal fluid of these patients contained more c3 than the cerebrospinal fluid of patients with cancer that do not have metastases at this site and that the amount of c3 that we find in the cerebrospinal fluid is proportional to the amount of tumor that we see using MRI, then we collect these cells from the cerebrospinal fluid of these patients and observe only in the cells cancer cells and we discovered that the cancer cells themselves are producing messenger RNA for jeet, the genes complement c3 and that they produce the gene product, as well as we measured it by immunofluorescence, so once we were satisfied that this was important in human diseases , so we established So we got to work and decided to figure out how this works.

We are going to find out the mechanism by which c3 allows cancer to grow in the cerebrospinal fluid, but this is a very complex problem and to really analyze this problem we need to think about it in a logical way and therefore the first thing we do with a complex system to be able to see which parts are necessary is to simply remove them, so in this case we used short hairpins to inhibit the expression of the c3 gene in our cancer cells and we found that inhibiting the expression of c3 inhibited the growth of the cancer cells at the base of the leptomeninges and in all of our mouse models, so that was a good clue and then we did the opposite: we took cancer cells that really don't have much of an ability to grow in this space the parental cells the cousins ​​of these leptomeninges cells that do not express c3 we added c3 back to these cells and found that it supported their growth, this way we showed that c3 was actually very important for cancer cell growth at the base of the leptomeninges, but it still didn't tell us how, so we gave step back and think about the space itself, so what is in the leptomeninges and in the cerebrospinal fluid and the leptomeninges and the cancer cells that are growing inside the base of the leptomeninges? spinal fluid, so this is the medium for your growth, but this is a terrible place to grow if you look at the spinal fluid, it has much less oxygen, much less protein or sugar, fats, iron, almost anything that is can imagine, much less. than the blood or any other part of the body, it's almost like cancer cells live on Gatorade instead of a good steak dinner and I don't know about you, but I prefer to have a steak for dinner, so I wondered how the cells could be living cancerous in this space.

Can the cells live in Gatorade or are they changing the way cerebrospinal fluid is formed? So we take a look at how cerebrospinal fluid is produced and how cerebrospinal fluid is produced by a The structure in the brain called the choroid plexus and the Koid plexus is a very small part of the brain, so this is actually a model of a life-size human brain and the choroid plexus lives deep inside. It's very, very small. There's a very small pink thing right here. So the Koid plexus that you can see here in an autopsy specimen is not very impressive, but when you take it out fresh it is a very beautiful structure.

I'm a big fan and it produces spinal fluid and the choroid plexus consists of a sac of epithelium. The cells and these cells have two functions, one is to secrete cerebrospinal fluid and the other is to bind together very tightly and prevent blood products from entering the cerebrospinal fluid so that it remains as pure and clear as you saw. This we took a look at the choroid plexus and looked at how complement c3 might alter the function of this structure in the brain. We discovered that the Koid plexus expresses the c3a receptor and then we thought that perhaps c3 could be the activation of this receptor. alters the function of the choroid plexus to make a long story short we thought maybe it changes the barrier function of this structure so we took a look at some dextran so the strands of Dec are our sugars of various sizes and the We label with fluorescence. fluorescently and distributed them throughout the mouse and then we took a look at the cerebrospinal fluid of that mouse before and after treating the mouse with c3, we found that when you treat the mouse with C 3 you can see leakage of some of these components from the plasma . in the cerebrospinal fluid and when we used a mouse that did not have a c3 receptor, a knockout mouse, this pattern did not occur, so this led us to the hypothesis that perhaps c3 a being in pinyin is altering the barrier function of the choroid plexus I took a look at the choid plexus under the microscope and you can see here this sort of green and red chicken wire arrangement.

These are the tight junctions that hold these cells together and when we treat the mite we treat the mouse with c3a. You can see that this becomes disorganized and this barrier is no longer functioning, so this tells us that our original idea was maybe a little simplistic that maybe we are not talking about spinal fluid that the cancer cells are alive that this is spinal fluid plus cancer and that the spinal cord fluid in the context of cancer is different and different enough to allow cancer cells to grow, so while it is not a meat dinner, it seems like something a little more sensible, this is enough for cancer continues to live, so if this if this.

It is true that if cancer cells can produce c3a and this alters the function of the choroid plexus epithelial cells, then one would imagine that if it were a mouse that did not have a c3a receptor, it would not be able to harbor cancer cell growth. in your cerebrospinal fluid and that is indeed the case, we found that to be true, furthermore, you would imagine that if you had access to medications that can alter the function of this receptor, you could alter the function of this barrier and perhaps also alter the cancer. cell growth and that is also true so we use c3 a receptor antagonist and an antagonist is a molecule that tickles a receptor but prevents it from working and when we do this it blocks the growth of cancer cells in this space and we use a c3 c3 an egg receptor and that's a molecule that tickles the receptor and allows it to function that opened up the blood CSF Berrien actually allowed the cancer cells to grow and this was true in all of our models so I hope this kind of brief, the story short leptin meningioma TAS ptosis shows that this iterative modeling approach of in vivo selection in mice and many other molecular worksreally careful not only show you a really clear physiology of the brain, but it also has the potential to help we help our patients and design new strategies for their treatment, and in case you don't, in case you feel a little Of suspicion about that, I'd like to take a moment to talk about parenchymal metastases or tissue metastases in the brain, so using these same approaches using iterative selection in vivo to generate metastatic cell lines, we were able to find interactions between the cells. cancer cells and another cell in the brain, astrocytes, so they are these types of red spiders that we find. that these interactions through gap junctions support the growth of cancer cells within this space and if we inhibit those interactions using commercially available drugs such as mclovin m8 we can inhibit the growth in a mouse model.

We have taken this approach to the clinic and we have been treating patients with it and I am just telling you the story of a patient who is 60 years old and has been treated with Brett. I am being treated for breast cancer for many years but her brain metastases kept coming back and unfortunately this is the pattern. I am treating this patient with this drug, we were able to inhibit the growth of these metastases so what we study is not just metastases In the brain, we study parenchymal metastases and leptomeninges, how we study them well, we observe, question and formulate hypotheses. and then of course we have to test our hypotheses and repeat them over and over again and the reason we study it is because we really want to help people and there are many people who have helped me so I have to thank my mentors .

Juwan and Lisa, as well as my collaborator at Elena and everyone who joined my lab, as well as all my funding sources. Thank you. Thanks Adrian for sharing that fantastic detective story. If anyone has any questions, come up to the microphone, that's the rule. above there is no additional loan hi so I think it's almost remarkable how we see that cancer is almost able to evolve and adapt its environment to ensure its growth and success so I was wondering have you considered it? how to apply this to like the phenomenon of angiogenesis in which cancer cells can force blood vessels to grow towards them.

You may like c3 because it can respond to the membrane and cause it to open, so have you considered applying it? the same approach to the receptors that angiogenesis binds to, so I guess those are great questions in the first place. Second, it turns out that the process of angiogenesis is a little different than the process I'm studying here in spinal fluid. that the choroid plexus, which is also known as the blood-brain barrier, is different from the blood-brain barrier, so the epithelial cells of the choroid plexus, those are the ones that form the barrier of the blood-brain barrier, are actually those tight junctions between those epithelial cells and not so much. a lot of the endothelial cells are more famous and involve angiogenesis, so in the case of leptin meningioma, maybe it doesn't help as much, but we certainly use those drugs in the clinic quite a bit, okay, sorry, I just had one more question , oh, cool, are you like um?

Investigating any other porn like quote C three in the body and seeing if we can almost inhibit the receptors that they bind to, so I guess so, we are looking at a lot of other responsible genes that could potentially support the growth of cancer cells in the CSF. or in the brain parenchyma, you definitely are, thanks here, how do you think the fact that the CSF does not have a microbiome that contributes to the development of cancer growth and likes metastasis? I think that's a great question, so what do I say in terms? of the microbiome, so looking at it is absolutely sterile, you're right and one of the things that I find so fascinating is that we don't fully understand the relationship of the immune system to the CSF, we don't fully understand how the immune system is able to survey this space and carry the antigen to the rest of the body so it can do its immunological job and I'm not entirely sure, but I suspect that when cancer solves this problem, part of the reason it can grow so quickly in the CSF is that it has adapted and has solved this problem.

Sorry, do you think it is also possible to create an artificial microbiome that would perhaps help fight cancer? I think it's a fun idea and yes, it is possible. I think we should try it, let's do it. I don't know okay we'll let Mary vada ask one question at a time so you're okay so I'm curious about the choroid plexus because its function is to protect against certain things enter the cerebrospinal fluid at the same time they produce the fluid and I realize it's quite similar to the placenta, so I'm wondering if you foresee any applicability of your research from an embryonic point of view, so I guess I'd say I agree that it's very similar. to the placenta, I think it's even more similar to the renal tubules, it's a structure that's actually so similar that they actually share a lot of genes in common and in that way I think a lot of this research really suggests that when you disrupt one of these layers of one of these stratified epithelial layers when you disturb one of these I'm... you're probably disturbing many of them within the system, so I think while I'm not sure how this would affect it would be important in development.

I know that the supplement performs many other functions in the body. Very well thank you. Let's move on here, so I know a lot of this work with c3 is heavily focused on features like cancer prevention. enter this region before you actually enter the region with c3, but after you have entered the region, it is possible to use the immune system to combat the here, which is an area of ​​active study, so currently there is a couple of clinical trials using some of these checkpoint inhibitors that you've heard about to treat leptomeninges doses anecdotally, they seem to sometimes work and sometimes they don't.sometimes and we don't fully understand why it works and doesn't work, so that you are right, let's go back to the other side, so you mentioned about 20 genes that are common among cells that may have the ability to go to a different environment and at the beginning you recognize that cancer cells mutate correctly, we always have to take into account So, does that mean that c3 specifically has the ability to mutate and create different variants of c3?

Well I guess. It could be, but what I'm talking about in terms of gene expression, it's an RNA type problem, it's a transcriptional problem, and in terms of mutation, it would be more of a DNA problem, so a lot of people have done quite a bit of work and are We're still working on this issue of whether or not changes at the DNA level could predispose a given cancer to metastasize to a particular site. So far we haven't found any genes or groups of genes that lead cancer cells to end up in the brain, but the other part of your question you were asking was: do you know that this gene could mutate?

Oh, it could, yeah, any gene definitely could, so if it could mutate correctly, one question we'll move on to the other side. because you'll get your answer if you cut the next one, we'll talk about how mutations create the energon a'ti that allows tumors to be different in the next talk, so let's move on to the next one here, so do it. you think I'm using genetic engineering, maybe through CRISPR would be a viable mechanism to eliminate the c3 AR or septic receptor. Yeah, I think it is though, just for full disclosure, you know that the c3 receptor is actually pretty important and the body. so these mice do well, but if you have a clear receptor in your body, if you don't have a functional clear receptor in your body, you're going to be very susceptible to infection.

One of the c3 functions of the c3 a receiver is permanent. aflatoxin, so it's your response a non-specific response to the infection, right, but you know, like many tools, the right tool in the right place at the right time. I don't question here. I like your seed and soil analogy, so I was wondering if c3 expression is essential for metastasis, not that this occurs only in tissues that express c3 expression, so what we found was that actually the Cancer cells need to transport their own c3, but by removing c3 from the system in our mouse models, it appears to be important in the mouse models.

I suspect you're probably right that once you know the spacing, I don't think it really matters to the cancer cell where the c3 comes from and what the blood-brain barrier is like. opened so I think you could make use of whatever you think is cool thank you very much Adrian well you guys are finally getting this now so I wanted to introduce you to an additional person who I will be emceeing tonight and that It's Dr. Michael Overholser so Michael get up yeah you need to meet Michael because he is the dean of our school so we have a school that is a graduate school so students come after we get our bachelor's degree and go to college and want to become biologists, they come here to train in cancer biology.

Michael is the principal of that school at any time. We have about 275 students getting their PhD here, but the most important thing is that you guys need to meet with him tonight. We also receive every summer about one hundred 150 students who are from the university who come to learn biology to learn how to identify the biology of cancer and below them, a group of approximately one hundred high school students who have the opportunity to come and work in the research labs here, so there are apps, if you're excited about this, you want to follow up on the questions you have and you want to see about it, it would be like working in a research lab or a research environment, already whether in the hospital or here in Michael's research laboratory.

The guy to contact, but also the brochures, will be in the back when you leave here tonight. He will finish being the master of ceremonies at the end of the day because unfortunately, after I came in, I introduce the next person that I will have to leave. Is it you, but have you been? a large audience and you asked a really important question that we want to end with, which is that you pointed out that cancers keep mutating, they keep trying different things and you asked how much does that contribute to the cancer's ability to metastasize when you asked Adrienne how much bypasses the immune system when you attract Prasad and so the real question is that's really fundamental to what we don't know about the biology of cancer, so this last year we were lucky enough to recruit one of the experts emerging in that field and that's our last speaker tomas tamela is a scientist who started his career in Finland, where he grew up, worked on the part of the metastasis problem that involved angiogenesis early in his career at Curie and then came to MIT to work. with one of the great people who developed mouse models so that we could study how cancer mutates over time and now we are lucky to have it here at Sloan-Kettering Memorial and take advantage of the tumors that are removed here in the operating rooms to understand them in a primary level and then take them and study them in mouse models and he's here to share his story for the last half hour here today so tell us okay thank you thank you for the introduction and it's It's great to be here with you this night and talk about some of the work that interests me in my lab and the title of my talk is decoding the complexity of the society of cancer cells and really the title has the idea that cancer cells actually work together they are like a family, they are actually all related to each other and that really creates problems because if you were trying to kill one type of cancer cell, you could get help for the lost neighbor, whatever this neighbor might be. help her voluntarily or just be there next to that cancer cell and then produce something that can really help your friend, so my colleagues presented very well the problem that we face when we treat cancer: it really is the problem of incomplete responses. in the clinic as dr.

Thompson mentioned at the beginning that if we can remove the tumor surgically we will be cured because we eliminate every cancer cell in the body, but if we can't do it if the cancer has already spread and we have to resort to anti-cancer drugs. and even some of these immune therapies that are very effective against cancer really have to go after every cancer cell down to the last cell until we can be sure that the patient is cured, so here what we are looking at is a lung cancer in and this is the type of cancer that I study in the laboratory, mouse lung cancers are inactually very similar to human lung cancers and the model that we are using is very relevant to human disease now if we treat this tumor and in this case, in this experiment, these tumors were treated with chemotherapy, a kind of cancer therapy more traditional called cisplatin, so initially when we treat these tumors, we see that the tumor responds, the tumor size reduces, so the baseline is here and now the tumor size reduces and the control mouse , the mouse that did not receive therapy, it is true that the tumor grew and now if we treat this mouse again with two more doses of chemotherapy we see that the response begins to attenuate, it is not like that. actually as prominent as it was in the first place and finally, by the time we get to the fifth dose and the sixth dose, this tumor has become completely resistant to the therapy, so this is really the problem and the way we think about it is that I have this tumor, lung cancer, in this case it is composed of cells that reside in different states, they can be genetically different or they can simply be in a different environment that makes them behave in a different way and when we treat this tumor that is very heterogeneous, so you have multiple different types of cells, we actually only attack a portion of those cells, only a portion of those cells are actually intrinsically sensitive to the therapy and some are intrinsically resistant and now, because we have these resistant cells , we allow these cells and we give the tumor as a whole time to develop what's called adaptive resistance and that's what we're seeing here, so how do we approach this.

The way we think about this in my lab is that heterogeneity in a tumor is a bad thing. The fact that the tumor cell society is very, very heterogeneous, cells behaving in a different way is a bad thing and actually the way to approach this is there are two ways that maybe we could do it and One of them would be to develop drugs, some type of therapy that would really push all these cancer cells, these cancer cells of different colors to this blue state and now this blue state, as we saw initially, would be sensitive to conventional therapy and that way would eradicate all of these cancer cells and the other way to do it would be to find avenues for other types of drugs that work in different ways and that actually attack those intrinsically resistant cells and now when we combine it with conventional therapies or immunotherapies or any therapy we want to use , we get a more complete response, so this would be great if we could do this, but it's actually a big challenge because from the beginning we don't really know which of these cancer cells respond. responsible for relapse which of these cells are actually intrinsically resistant we know that at least one of these guys is guilty maybe even several we know, but we don't know who they are, so fortunately when I was a postdoc at MIT there were two things that happened were really exciting, one of them was CRISPR, which I'm sure you all know, but the other was a technology called single-cell mRNA sequencing and what this technology actually allows us to do is something really cool, so just imagine. that you go and buy a smoothie that you really like and you want to make that smoothie at home you want to know exactly what it is made of if you could analyze that smoothie in some way and know exactly in what proportion and in what Unfortunately, I can't do that, we don't have a technology for that in the lab, but single-cell mRNA sequencing actually allows us to do something very similar for cancer, so we can take this tumor dissociated into enzymes. solutions so that we have a suspension of single cells and then subject them to a single sequencing, sorry, so how does it work?

This is a technology that has been developed and continues to be developed in many laboratories around the world. We are not the developers of this technology we are users of this technology and the way it works is basically, you have this micro Fulop fluidic system where the cells fly in from the side and then a channel of liquid here contains little beads that have little pieces of DNA in them. that capture any mRNA molecule in the cell and what this device does is it actually combines one of these beads and a cell into a single droplet and what then happens in that droplet is that the cell breaks down, is destroyed or is destroyed. opened in that droplet and now all the mRNA molecules inside the cell will be captured by the bead and each bead actually contains a unique barcode so we can read it in the mRNA sequencing reaction and what this allows us to do is build These types of graphs we call t-sne graphs and these graphs are actually basically we replace cells so that each point, each point here is a single cell and that point that cell is placed here on the graph based on how similar it is. to others can other cells, so here, for example, we have this or here we have this green group that is highlighted because all the cells that were in that analysis were similar to each other and then this orange group here is very far from that green, so these cells are very different from each other, so now if we do that experiment in this case, this is exactly what we did with our lung tumors.

It maps lung tumors, so we ground them up, put them through single-cell mRNA sequencing, and here we actually combined them. cells from two different mice, two different mice with tumors and you can see that these cells are placed in this group on this graph something like this. Now, in the next iteration, we took two different mice and two more mice that we had treated with cisplatin chemotherapy and now we can see how those cells are different and what we're actually seeing here is that clearly there are cell states or cancer cells here circled above that are preferentially depleted in cisplatin-treated tumors, so these are the cells that are intrinsically sensitive, these are the ones that are easy to kill with cisplatin therapy, while any cells that reside in This state has this phenotype that I've circled in purple is actually intrinsically resistant, so now this makes the problem a little bit easier, we go from a total of then 10 different subpopulations of cells to about five, so that now we have to figure out what drives the resistance in those five populations and to do that I have to take a step back and review some things that we think or know about it. can make a cell different in a tissue, so one of the things that dr.

Thompson alluded, of course, to the DNA of the cell, although all cancer cells within a tumor are related to each other, their genomes can be different because these tumors continue to mutate, so each cell or at least clones of cells can contain different types of mutations and therefore express or stop expressing certain sets of genes, then we have epigenetic modifications of DNA, just the axis from DNA to gene expression machinery can be surprisingly different in different cells, so these are what we call cellular intrinsic factors, in addition to the ones we have, we have all this large number of different cellular extrinsic factors, we have the availability of oxygen, the presence of nutrients, as Adrienne mentioned, the CSF fluid is very depleted in nutrients, so that is a very different environment for a cell that causes that cell to behave in a very different way.

We have other cells in these cancers that are normal cells, such as the blood vessels that form the angiogenic vasculature, the cancer cells come into contact with. with these cells in these cells they are talking to each other, they do not exist in isolation, they always interact, also, as I mentioned, the cancer cell itself, the cancer, its own cells form a society, they act together, they work together, they cooperate, too they compete with each other. Another one is the extracellular matrix, so the proteins that the cells adhere to can be very different from each other and in addition, we have these various changes in the biophysical environment, including pH osmolality and even stiffness and pressure of the fabric, all of these.

The factors act on a cell and that creates a huge amount of complexity, so how can we understand that it really is a simple equation or at least in this graph a simple equation? Whatever the state of the cell is, it is the sum of the cell's intrinsic factors. and then also the extrinsic factors of the cell that act on that and unfortunately, even just doing this in mind, we can do a single cell mRNA sequencing experiment because we now have a lot of information about the role of the various genes that cells express to infer.

Much of this information is based on the gene expression signature, so here is our data on lung cancer. This is the same data I showed you earlier, displayed in a different way. In this case, we have color coded the cells we think or actually do. When we ask the computer to do this, things are similar, so now we have in this case about 14 different types of cancer cells that form lung cancers in mice and now we can learn a lot about the state of the cells. cells and When we learn about the state of the cell, we can now start to think about ways to change the behavior of the cell, the phenotype of the cell or its state, so we would like to get these cells out of this state which makes us a state of proliferation and also cisplatin resistant to a state in which the cell stops proliferating and becomes insensitive to this plant, that would be an ideal response and to do so we would have to modify these Journal entries or modify the cell itself and today only I'm going to give you an example of something I worked on while I was a postdoc at MIT, which is a contribution to the cell and which actually involves the interaction between one type of cancer cell and another type of cancer cell.

Okay, so how do we get to this? We take this one. cellular data and analyzed them in various ways and also through them through many other types of experiments found and at home about this gene that has a curious name, it is called porcupine, it is because this gene was originally discovered in flies of the fruit that when you mutate this gene the fly looks like a porcupine the fly people have a very good sense of humor when they name genes, so even in mice and people, this gene still has the same name, so when we look at the expression of the porcupine gene, anything you read here is a cell that expresses porcupine, so this gene is present and just as you see here in this graph only some of the groups express it and then of course we had to go to the tumors and validate that this expression is actually true by staining the protein and here in red we can see the porcupine protein and now the really exciting thing about this gene is that it is actually associated with the secretion of a secreted signaling molecule called wynt And now We knew that these cells are probably producing Wynt, so we did another type of analysis, another type of staining experiment to identify the cells that activate the wind signaling pathway in response to that wind signal and here's when we do the analysis. and we really get close to these tumors, we find that always next to a cell that has activated the wind pathway there is no porcupine positive cell that is providing that wind signal, so now we have a hypothesis that maybe this pathway signaling is really important in generating heterogeneity and if we get rid of that wind signal, what could happen?

Now we reduce the heterogeneity in the tumor, so that's exactly what we're trying to do. We use a drug developed by Novartis and the pharmaceutical company. This is a drug that is now being tested on people actually and what this drug does is block the wind signaling pathway and if we treat animals now, these are lung tumor carrying mice with tumors with this drug LG k97 4 , you can see that the tumor growth actually stops the cells. proliferating, so we already achieved one of these goals that we wanted to achieve: we wanted to silence the cell and we wanted it to stop proliferating, so what about the sensitivity to cisplatin here?

The data wasn't as clear, but it's still very clear that if you look at what's called a waterfall chart, anything below this axis here is that the tumor actually shrank in turn in response to the therapy if we are in these groups where we combine cisplatin chemotherapy with an inhibitor that we actually receive. much better answers, so now we check both boxes at least to some extent, so this study we think serves as a model for how we can develop new types of cancer therapies that are conceptually different rather than tryingdirectly kill a cell. which is very difficult without trying to damage normal cells, what we can do in the future is try to push the cancer cells to a state that is more sensitive to therapy, so to summarize, let's get back to this.

The problem really is the heterogeneity that exists in these tumors and that can arise for many different reasons, but by doing these types of experiments where we use single RNA sequencing and other types of approaches, we can find out what these different are. States are and study their contribution to tumor progression, metastasis and resistance to treatment, and what we achieved in this study that I used as an example was to target one of these populations, so by using a Wynt inhibitor we are actually getting rid of these purple cells that we already knew were resistant to cisplatin and then when we combined this why not inhibitor with cisplatin-based chemotherapy, we actually improved the outcome in these mice and hopefully in the future in people as well , so I want to thank you for your attention and I want to thank my lab here, we are doing experiments and all my mentors and collaborators, thank you, how about you forget some questions for dr.

Tamela and then we'll open it up and bring in all the speakers and see if we can stump the experts, so first some questions for dr. Tamela hello, thank you very much for your presentation. I noticed on the last slide that the inhibitor blocked the signaling pathway, which eliminated most of the differences in different sectors of the population. What are other ways besides just pointing that out? can you inhibit heterogeneity in cells, yes, well that is a very good question, since I mentioned that heterogeneity can arise from so many types of stimuli in the cell and also from modifications that occur within the cell, such as epigenetic modifications or mutations, these days they were actually improving. and better to impact the epigenetic state with drugs the genetic state that is very difficult to fix once the gene is broken once the genome has mutated it is difficult to fix it again therapeutically certainly and when it comes to many of these things externally there are ways to drug them using inhibitors, small molecules or antibody molecules or even activating certain pathways if we wanted, okay, thanks, so your main goal in this is to transfer all the cells of a certain tumor to the same state, but so far.

You've focused on one that uses cell signaling to almost coerce most cells into the same state. Would it be possible to use other methods, such as treating a cancer, with a certain drug that does not promote certain states and for most other states and then cycle through multiple states of this to try to cover as many different types of cell states to try to create a treatment that stops the cycle, I guess, of having to do this over and over again or is that it? like they've seen it at all or for sure, that's it, that's a great point, it's actually exactly those are the exact types of experiments that we're doing right now in the lab, so what we're doing is combining, like this that every time we do a single-cell RNA sequencing experiment. for example, or any type of tumor analysis, we're just capturing a point in time, a snapshot of what that tumor was at the time we removed it while we're trying to do in the lab, we're actually trying to create systems that allow us to record the history of the cancer cells within their DNA, actually using CRISPR, it's called CRISPR genome recording, it affects it and combines it with the RNA sequencing data and that way we will find out how those states are changing dynamically, amazing, thank you, hi .

Has there ever been a scenario where a new species of tumor cells grew while you were sequencing the old set of tumor species? Yes, that's a very good question, what we haven't really looked at much is the adaptive part of tumor heterogeneity, so I think what we mean is that we have the best opportunity to attack the tumor from the beginning and then that some cells survive that initial blow, even if it was a very hard blow. We kill 99.9% of those cancer cells. If we have that little action of the remaining cancer cells, what comes out will probably be very different from what we had at the beginning.

We haven't done those experiments, but other people have and discovered this. actually thank you oh hi um I'm not sure about this I feel like this could actually create cancer cells but I was wondering if you could use um sorry um could you use uh I went to like can you give I went to I like it, regular? cells or natural cells to prevent the side effects that come from chemotherapy to prevent them from losing their genetic material and you know, baldness and things like that, that's an amazing question and you're actually right, this is true, so there are experiments that are not carried out in my laboratory but in other laboratories that have shown that some of the side effects of radiotherapy, as dr.

Thompson said that at first they normally affect tissues that have proliferated a lot and one of those tissues is the intestine and it turns out that if you apply it if you activate the wind pathway in the intestines, this has only been done in mice and then treatment is provided. With radiation, the wind actually protects the gut very well, so you're absolutely right that that would be one way to do it. It would be difficult to do it in people. I think restrict it specifically to the gut, but at least in you know the proof of concept is there, thank you, okay, let's get one more and let's not go anywhere, but then I'll bring the other panelists or other speakers here as well, so keep your questions and we'll try to get some. more for any of the speakers, okay, but one more first, okay, so above all, you should know about mutations and what the spontaneous growth of cancer looks like, once we reach that state of cancer cells where are homogenized in a type of population, considering time, especially, what?

Is that window or how long-term is it a solution considering that more mutations of the homogenized population are not mitigated? Yeah, so I guess what I showed you is real. No, I don't want to say it's a pipe dream, but it's very difficult to do, but it just describes the composition, the concept of where we're trying to take this and yeah, what you're saying is: do you know how long we can stay holding the cancer cells in one? The status is probably not very long, even if we could do that, then we would have to immediately continue with the therapy to really determine the timing of how these therapies would be designed in the future, that is another big challenge. but we're really in the early days of this right now thank you okay don't go anywhere.

I just want to invite Adrienne and Prasad here real quick. What happened to your question? It went well. Get in line. I wanted you to do it. Stay, what is this? Okay, is everyone feeling smart? You look smart. It doesn't sound like you feel smart. This side of the room is proving it. I'm not sure about this side. What's going on here? No, it's too long, so in graduate school. school in grad school your average class is three hours long sure okay so don't get tired of us one more time y'all feel smart now is your chance yeah so this is what dr.

Thompson alluded to seeing if we can stump the panel so I want the remaining questions for dr. Tomas, but also for any of the speakers today, ask us some good questions, so he mentioned in his speech how he could edit the input and other factors like pH and just the environment in general. Has he ever tried the other factors and if he has ever taken the risk of changing the shape of the ordinal chamber, as if only the genes of the body's cells in general set the therapy or other treatment methods? I am actually harming the body because the cells are now more sensitive. about whether the treatment is yes, as you said, there are many things and, as I showed on that slide, that impact the state of the cells, some of them are easier for us to study, even doing this a little bit.

A portion of that Complexity took a long time, so we haven't been able to look very comprehensively at all of those things at the same time, but today it's getting faster and easier to do those experiments because we have this unique technology. cellular sequencing where we can look at all the genes that are expressed in the cell and infer those various relationships and then try to manipulate them experimentally, that's the ultimate goal, but we haven't been able to do it. I have a question. In Prasad's case, you mentioned how safety cells or safety genes are put into T cells, but I was wondering if cancer cells have ever mutated to exploit safety genes.

In theory, it might be possible, but again, when we do genetic engineering, that's the different part of the process. science where we know exactly where to insert to prevent that or reduce the possibility, but having said that in the clinical trials we continue to make sure that things don't happen to understand thank you. I also have a question about match cell therapy. I'm wondering what the benefits and drawbacks would be of injecting auto T cells into babies even before they have cancer, so I wish that by the time you get to medical school and science we've moved on so far, so initially we're doing the immunotherapy .

We tried it with advanced cancers, but the thought process is: why don't we go to early-stage cancers? So those clinical trials are ongoing now, for example, at Memorial Sloan-Kettering, we did a clinical trial on patients with stage 2 and 3 lung cancer. You used to give chemotherapy for the reasons you heard and then have surgery and take part of the lung where the tumor is. So what we did was give two doses of immunotherapy while the cancer is there, educate the immune system, stimulate the immune system to find. the cancer and then we went to the operating room and they removed it to our own surprise, 40 percent of the patients had what we call a complete response, obviously we have to wait and see how they do in the long term, so slowly the immunotherapy in which we are.

By reaching the late stage, we are moving to the early stage and the next step will be prevention, so that is what science is heading towards. Thanks, let's get another one on this side for my friend. I brought backup and then come here, okay, continue with our question for mr. Tamela, so when you said that you like to plot all the cells based on how similar they are, you also said that there are many factors that play a role in how similar the cells are, so how do you like to plot them all? this on a two dimensional graph like your diagram was like I think it had dimensions 1 and 2 for the axis Oh, uh, and I heard you had like um I mean, I heard you say that you came up with an equation for like it's like a sum of all the factors, so how specific is this, yeah, those are both amazing questions first of all, so you're absolutely right in doing those plots that I showed that in two dimensions took many, many years to degrade.

It took advanced mathematicians and computer scientists to develop those analyses, and you're absolutely right, we have about 25,000 genes in our bodies and each cell can express itself, it's usually expressed in thousands of them and they can go up or up. in a cell, so it actually has thousands and thousands of dimensions that are being recorded in this analysis and what this two-dimensional analysis or plot really is is simply forcing all of those dimensions to two dimensions, so it's a reduction of dimensionality. analysis and that way it's not really a linear process, you're losing information as you do it and those distances between those different cells when you look at them on the graph, they're actually arbitrary distances, so it's a good point and so on and so forth. then yes, some of the intrinsic and extrinsic is the cellular state, which is largely an oversimplification, we infer a lot of it from the transcriptomic data we obtain, we make assumptions and conclusions based on our prior knowledge of various genes, the role of different genes and programs, and we're just looking at the most important things that happen in these cells, so that's the level we're at, but those things are the most important, usually, more or less. we're learning real biology from them, thank you, okay, we have four more, we'll get through them and then I want to invite anyone who has questions that they've been holding on to or aren't sure about, to come forward. and talk to whoever you want when we're done, that's fine, but we have to finish it soon.

I know some people have to move on, but we have four more and then we'll close it and you could. welcome to come tofront, so on this side I have a question for mr. It sucks, sorry, I understand one of your articles says you found that localized injection of cheese sauce for cars was better than Lee's intravenous injection, so do you think car T cells could combine it with nanotechnology to create types ? intravenous injection is better, I think both or we need to study well. I mean, we're injecting the car T cells directly into the tumor in the mouse experiments, so when I was interviewing a postdoctoral candidate like you, I asked, "Okay, you're injecting there." but what are the corpses doing?

Are they taking blood vessels? Are they going to what is called stroma? What are the roads for car T cells? Are they taking the lymphatic vessels? How are they entering the tumor? Well, I'm putting it right next to the tumor, but how they go into the tumor and, surprisingly, T cells by nature like to circulate through the body and we noticed that in the mice within a day, even though we inject, we see in the peripheral blood in the clinical trial that we are noticing. The same thing, although we inject karpizzle directly next to the tumor, they recognize that they are activated within a day.

I can see it in the patients' peripheral blood when we do the testing, so now we've taken those Kotb cells and we're trying to The study and what we're learning at Lee is very preliminary once they get into the tumor, once they're educated , now it is better for them to enter a second site because now they are educated, but having said that, they may also all run out. and so on, so the ultimate goal is maybe we can stimulate by local injection, but the ultimate goal is to buy through the systemic system because that's where we can go to every organ and every site.

Thank you, this question is also for dr. Prasad um, you were mentioning how T cells become depleted and you have to keep modifying them and keep changing them genetically so that they can fight cancer cells. Is there perhaps a way to avoid such a constant fight for T cells? cells and maybe go directly to the cancer cell itself and maybe add something to the outside like a receptor or maybe genetically change the cancer cells, yes, the cancer cell itself, so maybe it starts fighting other cancer cells in Instead of T cells, yes, that's where I'm looking to my two colleagues for how to manipulate cancer cells, how to label them, how to identify them, how to know where they are, that's where we work collaboratively and that's where I'm looking for their knowledge on how to label cancer cells and how to find them, that's a great idea, but maybe you can answer, yes, yes, so cancer cells are very good at hiding, that's one of the main clinical problems, unfortunately, you know, in our mice , my mouse models have luciferase and are shiny. elements, but that's not actually how they work in real life, a big part of cancer research is identifying something called biomarkers, something we can use to find the tumor when we can't find it detected by other means and partly . part of that is just the heterogeneity of the tumor itself and the kind of genetic slippage of the slipperiness of the tumor, it's going to continue to mutate and alter its transcriptome, so it's a little difficult actually, thank you, okay, so I just have one question this time but it's for dr.

Adrienne and the question is does c3 affect the brain's ability to produce hormones effectively, so I know that c3 affects, it goes into the leptomeninges space, but also the leptin meningeal space is only a small part of the brain, so what that would affect other parts of the brain. The brain produces hormones that are sent throughout the body, so it's an exceptionally smart question. First of all, yes, the brain produces some hormones, from the hypothalamus to the pituitary, and those parts of the brain are some of them known. like the circumventricular organs, which means they have a kind of unique relationship with the circulation, so they don't actually live in the maninjau leptin space, they live behind the blood-brain barrier, but you're right, they do have a unique relationship with the Bloods that allows the hormones to come out and the brain to receive information to the extent that it wastes the functions of c3 in that system that is unknown, we know that therefore the complement c3 is actually divided into c3a, which is what I was talking about and then there is also c3 be c3 be inside the brain it is incredibly important for development, it helps with neuronal pruning and with and only with the development of the brain, so you are right, the complement is a protein very powerful, thank you for this.

The question is also directed to dr. adreno about when you and your side in the box you have you, which included c3, there are other genes in that box as well, what were the results with those with those genes similar to c3, so are we working on it? working on this second gene which is lipo Kalyan - LCN - we are working on that right now in my lab and it looks very promising so you are on the right track, thank you, okay, thanks for the questions, they are amazing, just a couple. of closing remarks and then we'll wrap it up and invite anyone who wants to come forward to ask questions.

I just want to say how amazing it is to see you all here, it's wonderful to see so many students interested in the basics. science and biomedical science at your age and you are all thinking at a really high level, it's amazing, this is the most exciting time to be doing science, we can edit the CRISPR genome, okay, we can know which cells are expressed at the single-cell level. At any given time in a population we can engineer cells and return them to the body after having tricked them into attacking cancer. It's amazing that some of you in this room, if you look around, make incredible discoveries in your lives. races, okay, I hope you hear things that we haven't even thought about, so we want you to come back, we want you to come back here to these events and learn how to make discoveries with us, okay, so thank you for coming tonight. our summer program hop for summer research check out the iPads in the lobby if you want to learn more about more programs one last time for our speakers and thank you okay.