 My name is Nick Stinson. I will co-facilitate the Waldenstroms, it's important for my wife Connie, she can't be here tonight so I'm filling in. Waldenstroms or WM is also a form of lymphoma, people may not know. Is there any WM people here tonight? So a couple of months ago we had a meeting and had a guy up Jack Whalen who introduced us to McKayla. I don't know how long you guys have known each other, but it was a little while. So yeah, it turns out McKayla is right in our backyard here in Portland at May Med, right? Doing research into lymphoma and the blood cancers. So fortunate together to come over and talk to us. Great, thank you so much Nick. Really appreciate that introduction. And thank you so much to the Cancer Community Center and for Molly and for you all for coming and letting me speak today. And feel free to interrupt me if you have any questions. And also if you can't hear me or anything doesn't make sense, I'm happy. We're being recorded too, but I think that that's fine. And Heather is my research associate in my lab. Heather Fairfield who actually got her master's degree and has worked for six years up in Jackson Lab. So she's one of the strongest researchers in my lab and I'm really lucky and happy to have her. And she's with me today to help us go through the slides. So I'm just going to start off saying a few weeks ago Senator Collins actually came down to May Medical Center and met with my mentor and I, Dr. Cliff Rosen, who actually knows her. And she's doing a lot of work down at the NIH to try and raise a lot of money for diseases for aging. She's very interested in Alzheimer's and aging diseases and also we're trying to raise money for cancer research. So a lot happens up at May Medical Center and a lot happens in the Research Institute. So I'm happy to share what my lab does. So go for it. So today I want to just tell you a little introduction about who I am and then I'll give you some background as to who my team is. And then I'll talk about the problem that my lab is focused on and we're going to talk about what is cancer in the bone marrow and what does it mean for cancer to grow in the bone marrow. And then we'll talk about the impact of this cancer. So what are the consequences when you have cancer that grows in the bone marrow? And then we'll talk about some of the solutions that are either being developed in my lab or being developed in other labs. And then we'll talk about specifically where do we go from here. So as you all know, this is really a cutting edge field. We don't have all the answers. We're just trying to piece together what other people have done, what other research labs have done and trying to take step forward so that we can have better treatments and hopefully cures one day for blood cancers. So this is my old lab and these are some other colleagues that I met at the American Association for Cancer Research. And so now I'll talk to you about my current lab. So my team is made up of some amazing scientists. I have one postdoc fellow. This is Caroline Fallink. She got her PhD from the University of Maine and also her master's degree. And she joined me in September. We all started on the same day. We already met Heather Fairfield, but she also has a master's degree from Wake Forest in North Carolina and worked at the Jackson Lab. So she's excellent with the mouse models that we really rely on in this disease. And then we also have a whole range of high school students and college interns who either joined for the academic or the summer. And they're pretty pivotal to help the research go forward as well. So this is my lab. And another key member of my lab is Dr. Cliff Rosen. So he's an MD who is a New England Journal of Medicine editor and he basically recruited me from Dana Farber and he helps advise and guide me in my research as well. So a lot of you already know a huge background about what is cancer, but I just thought we would start very broad. So in general, what is cancer? So cancer is a type of cell that has been mutated. So it was a normal cell that obtained a mutation that inactivated one of the tumor suppressor genes in the cell. And this basically makes it replicate and grow without stopping. These mutations can then lead to a whole bunch of other types of mutations. So there can be mutations that stop DNA repair. They can be mutations that are a proto-oncho gene, which is basically a gene that can then start cancer. And they can be all sorts of other types of inactivation of tumor suppressor genes. So normally tumor suppressor genes stop tumors from growing. If these types of genes in your DNA get a mutation, all of a sudden that normal cell that would be suppressed starts to grow out of control. And this leads to cancer. So Robert Weinberg and Dr. Douglas Hanahan actually developed the hallmarks of cancer in the year 2000 and they published this. And these hallmarks of cancer up here. And the first one is actually sustaining proliferative signaling. So basically as we talked about this is when tumor cells proliferate or they grow without stopping. And the second one is their ability to evade these growth suppressors. So naturally in all of our cells we have basically tumor suppressors that tell cells to stop growing. And that's why we don't have lots of cells growing out of control. But tumor cells are able to evade this. The third one is that these tumor cells not only grow, that would be okay. But they invade and they metastasize and they spread throughout the body. So they have these mutations that allow them to transfer from one location to other locations throughout the body. The fourth hallmark of cancer is the ability for tumor cells to enable this replicative immortality. So basically normal cells when they get these issues with their DNA this allows them to stop growing. But tumor cells take these mutations or nicks in the DNA and they actually are able to overcome this and continue to grow. The fifth hallmark is inducing angiogenesis. So this is the ability for a tumor cell to not only grow but then pull in blood vessels from the surrounding microenvironment. Which just means the surrounding cells. So normally a tumor cell might grow to a certain size and if it doesn't pull in blood vessel cells it will stop growing. Because it needs oxygen and it needs nutrients to continue to grow. But if that tumor cell is able to produce factors that tell blood vessels grow into the tumor then this tumor is able to then continue to grow larger and larger. And the sixth hallmark is resisting cell death. So this is similar to what we talked about before. But tumor cells or normal cells often go through something called apoptosis which is induced cell death. And this is a very normal good process in your body. But tumor cells are able to sidestep this and basically when they should be dead they continue to grow. So in the year 2011 they actually added four more hallmarks of cancer. Which I think is pretty interesting to consider that in 11 years we developed four new traits or properties of tumor cells. But these had previously not been recognized as important targetable factors in cancer. So I guess what I just take from this is that we're still learning more and more about how cancer grows. And this means that we're still able to come up with new ways to attack the cancer. So all of these ways are potential vulnerabilities, potential ways that you could go after the cancer. But there are also potential ways for the tumor to evade therapies. So it's really a very complicated process that means we have to use often lots of different types of drugs and target it from different angles. Because tumor has all these different ways to adapt to the drugs and the radiation. So the first one of the new emerging targets actually is this deregulating cellular energetics. So what this means is that cells are able to change their metabolism. So some cells grow with fatty acids or lipids. So when you eat fat the fats distribute throughout your body. And some cells take in those fat and they use the fat as an energy source. But other cells take in glucose or sugar and other cells take in protein and that becomes their energy source. And all these energy sources are useful for all of our cells in our body. But we're starting to learn that tumor cells actually have a different metabolism than normal cells. And we're still trying to understand what it is and what the differences are in different types of tumor. Because each different type of tumor has a different type of metabolic shift or change. But once we understand that better we might be able to change the food and the energy source for each tumor cell. And now people are actually getting more into fasting and trying to starve the tumor in different ways. And see if you can change the energy source for the tumor to try and change how the tumor grows. I think this is a really interesting field as well. We were just talking about this one today. So avoiding the immune destruction. So we know that normally your T cells and B cells and natural killer cells are in your body to keep bacteria and fungus and virus at bay. And they also kill tumor cells. The problem is that tumor cells are able to produce proteins that basically suppress the immune system. They keep those B cells and T cells that are supposed to be killing the tumor. They keep them from killing the tumor. So in this way we call this either evasion of the immune system or immunosuppression. Basically the tumor cell is able to escape the normal attack from the immune system. And this is a really hot topic right now. I'm not studying that very much but I know that it's a growing field and we're learning more and more. We learn more about the immune system. We learn more about what cancer is doing in the immune system. The ninth one is tumor promoting inflammation. So we know that inflammation in a lot of ways can cause reactive oxygen species. And that can lead to mutations in your DNA. And this inflammation can also relate to changes in your immune system. And as the tumor grows it also can produce inflammation. It's sort of an inflammatory location. So there's a lot that we're learning more about how inflammation works. What are the proteins involved in inflammation and how can we target this? And the last one is genomic instability and mutation. Which is basically again how tumor cells are able to survive. When a normal cell gets mutated it usually dies but tumor cells are able to take these mutations and then take more and more mutations on and not ever die. So that was an overview of all cancers. So what we're studying in my lab is blood cancer. And that's why I think a lot of you are here and are interested. So just to give you an overview of the different types of blood cancer. There's leukemia and there's lymphoma. And there's a few different types of leukemia, ALL, AML, CLL and CML. Which are all mutations of a type of blood cell. Similarly lymphoma is also a mutation or a cancer of a blood cell. And this grows in your lymphatic system. Leukemia typically grows in your circulation. So in your white blood cells that are circulating through your blood. But they're both a type of cancer of your blood cell, your white blood cell. But the next one is what we study which is called multiple myeloma. And also Walden-Stromes macroglobulinemia also falls within this category. So there's so many different details even within blood cancer that we kind of have to focus. So my lab is really focused on understanding myeloma. And I'll explain more the difference between that. But myeloma is the second most prevalent blood cancer after non-Hodgkin's lymphoma. And in 2015 accounted for 11,240 deaths in the USA alone. And right now there is no cure. There are good treatments. They can give people years and years on their life. But we're looking for better treatments and hopefully a cure one day. So what is multiple myeloma? So I think that this schematic helps me understand it the best. So basically in your blood all your blood cells come from one stem cell. Basically this stem cell right here. This multi-potent hematopoietic stem cell. And as this stem cell divides and changes it's able to turn into different types of sub-specialized cells. So one of them is the common myeloid progenitor. And the other one is the common lymphoid progenitor. And this is important when you're thinking about either lymphoid cancer or a myeloid like CML versus CLL cancer. So those are the two major families. And then within the lymphoid progenitor there's natural killer cells. And then there's these lymphocytes which are T cells or T lymphocytes versus B lymphocytes. And then if you go one step smaller you'll see that more specifically a type of B cell which is a fully mature B cell that's able to produce antibodies is called a plasma cell. And you and I have these and these are good. And they keep us healthy by producing antibodies against all of the different things that are infecting us every day. But the problem is when this plasma cell gets those mutations, gets those hallmarks of cancer and is able to grow out of control then we call this plasma cell a myeloma cell. And we call it multiple myeloma when the myeloma cell grows not just in one location but it actually grows in different locations throughout all of your bone marrow. So that's the disease that we're really trying to focus on in my lab. And just to give you an image of what this looks like these are all different myeloma cells. So there's the nucleus and this is the cytoplasm of the cell. So there's a few other different cancers that grow in the bone. So my PhD was actually in breast cancer bone metastasis. So we studied how breast cancer traveled from the breast through the circulation and got into the bone. And there's actually a lot of similarities between how breast cancer grows in the bone and how myeloma grows in the bone. They both get to the bone marrow and then they both start to mess up this equilibrium in your bone that keeps your bone healthy. So I'll talk about that in a minute but I think it's important when people talk about bone cancer to realize that there's different types of bone cancer. So there's osteosarcoma which is basically a mutation in a bone cell. So in an osteoblast you can actually have a cancer that starts in the bone in the hard part of your bone and then develops into a cancer there. But more commonly are these secondary bone cancers which are basically metastasis. So the mutation started in a prostate cell or a breast cell or a skin cell or a long cell and then they start to mutate and they leave and they go through your circulation and they end up in your bone marrow. And your bone marrow for a number of different reasons is very hospitable and very welcoming to tumor cells. So we're trying to understand why is the bone marrow so welcoming for these tumor cells and how does it support them. So it's very similar to breast cancer and we'll talk about that a little bit more but I think it's important to understand what is cancer in the bone. So myeloma would fit sort of in between. It starts in the bone marrow but also spreads to other bone marrow locations. So even if the first tumor cell started in your leg you might actually have metastasis or spreading from your leg to your arm. So probably you all know this even better than I do but what are the consequences of these types of cancer? Well one of the biggest consequences is that myeloma induces something called osteolytic bone disease. So osteolysis is when your bone gets degraded. So as the cancer grows in the bone marrow it starts to activate the cells in your bone that eat up the bone. They release the bone, the calcium that keeps your bone strong are released from the bone. So what does this mean? This leads to all sorts of problems. So you can see here that there's some punched out lesions. It basically looks like somebody just punched out parts of your bone. There's all sorts of different types of lesions but this one is very common. This happens in the skull, in the long bone basically anywhere that there's bone marrow. In the vertebra this is a big problem. So for example here's a tumor that's growing in the spine or in the vertebra next to the spinal cord. So consequences of this are shown here. So we have nerve compression, fracture, pain, your bone is weaker. This can lead to hypercalcemia. So as your bone gets degraded the calcium starts to spread throughout your circulation and this can lead to all sorts of problems for your organs. And this leads to impaired immune function. So as your normal bone marrow where your blood is made gets packed with tumor cells your blood can't make more blood cells. Your bone marrow can't make blood cells the way it normally should. So this is what we're trying to stop. So one analogy that they say, and this is hopefully helpful for you is to understand why we're studying the bone marrow is that we try to understand the soil. So how can we target myeloma? Well one way we can target them is not by going after the seed. If it was the same seed that was planted in this rainforest and in this desert they obviously didn't both grow the same way. Just like in your garden at home you need the right soil. So different seeds in different types of soil will grow well. Some grow well in one soil, some grow well in a different soil. We're trying to understand how these seeds, these tumor cells that are circulating stick in the soil of your bone marrow. Why do they grow so well here? And what can we do to make your bone marrow less hospitable and less welcoming? And this is a very useful way to target cancer because in myeloma there's a lot of different mutations. So there's no silver bullet. We don't have in myeloma one curing drug or in Waldenstroms or in CLL because there's a lot of different types of mutations. So instead of trying to get a drug to fix that mutation, get a drug to target that mutation, if we can go after the commonality between all of these cancer cells which is that they grow in the bone marrow then we can hopefully stop all of the cancers that grow in the bone marrow. That's my goal, one day to stop all of the seeds, all the tumor cells that get to the bone marrow from growing there. That would give people, that would be amazing. So exactly, this is the same seed that's planted in different soil. So hopefully that's a good analogy when you think of how tumor cells grow in the bone marrow. So just to give some credit, this hypothesis was actually first suggested for cancer in the 1880s. So this was Dr. Stephen Padgett described this as his seed and soil hypothesis. So people used to just be targeting the seed. They said we have to kill the tumor cell and you'll have to go after the tumor cell but now we're starting to understand it's not just the tumor cell. If you can understand why and how the tumor cell is growing within the context of all the other cells and proteins that it's surrounded by those might actually give you new clues and new ways to kill the tumor cell or at least stop it from growing. So in the bone, this is just to show that it's a very complex environment. Bone responds, bone is a very dynamic responsive organ. It's not just a static organ. Bone actually has cells inside it that are responding to all sorts of different hormones and forces and prostaglandins and vitamins, vitamin C and calcium. And you have two different types of bone. You have cortical bone which is on the outside of your bone. That's the hard part of your bone. And then inside your bone which makes up 20% of your bone mass is something called trabecular bone. And that's what people call spongy bone. It's not really spongy but it kind of looks like a sponge. So most of your weight is really held up by your cortical bone but your trabecular bone also gives your bone some ability to take some force and not be too brittle. So myoma plasma cells very much prefer to grow in the bone marrow. So if we can understand the complexities of this bone marrow, like I said, we can try and understand how does the bone marrow induce this anti-apoptotic property which means it stops the cancer from undergoing apoptosis, which is cell death. So we're trying to understand that and how does the bone marrow induce survival of the tumor cells. So, yes. Is the bone marrow within the trabecular part of the bone marrow? Yeah, actually, I wish I had a good picture but pretty much the bone marrow fills up all of this inside canal. So if you've ever had some ribs or a steak or anything, if you look right down the middle of your long bones, it's a hollow cavity that has your bone marrow in it. And yes, it interacts, especially in the end, there's a lot of trabecular bone that's surrounded by this bone marrow. The cortical bone is pretty much a solid. It has some cells in it, but there's not a lot of bone marrow. So it's kind of between the ends of your bone. And that's exactly where the cancer is growing. So this is very complicated. The point of this slide is that there's a whole bunch of different cells in your bone marrow. So you don't have to know what they all are, but not only are there different types of cells in your bone marrow, there's different types of chemicals in your bone marrow and proteins that are keeping you healthy. And there's all these other types of extracellular matrix components. So it's difficult to know exactly what to target, but it's really important to understand how all these pieces and parts fit together and what protein interacts with what other protein and what cell interacts with what other cell to really understand the biology of the bone marrow. Because before we can understand how cancer is growing there and how it's messing up the bone, we really have to understand the bone marrow. So in my lab, what we've been trying to do is get a better understanding of how myeloma grows in this very complicated environment. And then how can we identify new vulnerabilities, so new targets? Because if we're going to come up with a better therapy, we need a better understanding of how the bone marrow works and what are the cells there and what are the proteins and what is a good target. So we have bad targets and we have good targets, meaning some things work and some things don't. And how can you get the most effective and the best target and the best pathway or some cell to go after? And that's what we're trying to understand. So since this is so complicated, we have to use models. We can't go into humans. We can't study everything in humans. We can't reproduce this in a dish very easily. So we use all sorts of different types of models. One is a mouse model where we give human cells or mouse cancer cells into a mouse, inject them in the mouse and then we study how it grows because the mouse is actually very similar and a lot of these bone marrow cells and proteins are actually the same in the mouse and the human. They're not all the same and we have to be careful when we try and take something that worked in a mouse and go to humans. But a lot of it's very similar. And the other thing that we can do is we can do cultures. So we can just pick out the dipocyte, which is the fat cell. I don't know if you knew, but you have fat cells right in the middle of your bone marrow. And we also have, this is trabecular bone and there's these osteoblasts in those build bone. So we can do cultures of those. We can pull out certain cells and some of them we can grow. And these can then help us to kind of study bit by bit, one piece at a time. And as we get better at this, we can put different bits together and we can understand what are the key factors in the most important cell. For example, the red blood cells, we usually don't really put in culture. We put in other cells, but the red blood cells are something that most people aren't really looking at. Anyway, the important thing is then we develop hypotheses. We think about what could be a way to stop cancer. What's a novel new idea? And this is based on literature and reading and talking to people and looking at articles and also thinking and kind of trying to be creative in your own lab. And then we try eventually, once we test these and we go back and forth a few times between targets and models and testing it and coming up with a new hypothesis and finding out something new. Eventually what we hope to do is translate this to the patients. So we work a lot with the doctors. They talk to us, we talk to them and we try and come up with ways to do clinical trials or smaller trials and see if we can actually get this to people who need it. So this is my goal and this is what's happening in most labs right now. So I think now we're just going to go a little bit more in depth about, I'm going to tell you about one experiment in my lab and it's kind of a big experiment, but I think it's really promising. This has been thrilling work for me. This is a collaboration with people in Australia. I think it's hot in here. Does anybody else? Can I open the door? I don't know if this will help, but also feel free to get a drink or some food if you want. So this collaboration has been building for years actually. And the people in the Garvin Institute in Sydney, Australia are my colleagues. So I have colleagues who I've worked on at Dana-Farber Cancer Institute and my colleagues at the main medical center. So we're all work, and Novartis Institute actually in Switzerland has been donating the antibody for this study. So the basic biological understanding of what happens on the cellular level is shown here. So as the myeloma cell grows in your bone marrow, it interacts with two major types of cells for this experiment and for this test. And one of them is called osteoblast and the other is called an osteoclast. So the osteoblasts sit along your bone, the edge of your bone, and they make new bone. So these cells put calcium and hydroxyapatite into the bone, and they build it up and they make a strong bone. And these are really important cells. But they are coupled to osteoclasts. And there's always a balance in all of your bone that keeps the bone cells that are building bone tightly coupled to these osteoclasts, which resort bone. So these osteoclasts have a ruffled border and they produce acid and this degrades your bone. And there's always a couple between these. So as they move along the side of your bone, one cell, the osteoclast eats up the bone, and then the osteoblast comes and lays new bone. The way they do this is through a very complicated relationship and communication that keeps a nice homeostasis. Now this is impaired in a lot of diseases like osteoporosis and other types of bone diseases. And in myeloma this is also impaired because basically this equilibrium between these two cells is broken. Myeloma cells induce osteoclasts to degrade bone and eat up bone and they inhibit osteoblasts. So they stop the cells that are supposed to make new bone. And this is one of the proteins that inhibits osteoblasts. And this is both produced by myeloma cells and now we're learning more and more. It's produced by other cells called osteocytes that sit in your bone. And this protein that inhibits osteoblasts, if we can get rid of this protein, we can get these bone cells to start working again and producing matrix and healing these big holes, these punched out lesions that we see in so many patients. So what this project is based on is the hypothesis that if we remove sclerostin and we stop it from working that we might be able to help the bones to come back. And actually there's a drug that we used in this study that's used for osteoporosis trials. So it has good translation ability. Hopefully it can come to patients. So again, this is a little complicated, but the only point is that not only are the osteoclasts activated and the osteoblasts inhibited, but as these osteoclasts are activated, they eat up the bone. What happens when they eat the bone? The bone is actually filled with growth factors and proteins and collagen and all sorts of good things that actually feed back these growth factors, all these little molecules feed back to the tumor cell and they make the tumor cell grow faster and faster. So what we call this is actually the vicious cycle because then as the tumor grows faster it makes more osteoclast activity and it resorts bone faster and as that resorts bone faster it feeds back to the tumor. So we call this the vicious cycle of bone destruction and a lot of drugs are actually looking at stopping osteoclasts. So this phosphonates, inhibits this, but we're trying to actually target the osteoblasts and make these cells, these osteoblasts come back to life and start to make new bone. So what are the current treatments? So antiresorptives, I don't know if you've heard of these. If you've had any bone disease, any cancer-induced bone disease you probably have. So these are antiresorptives, meaning they stop bone resorption or eating away. So they stop the osteoclasts from eating up your bone and these are considered standard of care for cancer-induced bone disease and they work very well to get rid of osteoclasts which eat the bone. But the problem is that these SREs or skeletal-related events like fracture still occur because even though the bone isn't being eaten away, those holes remain. Those holes in the bone remain. So we need to try and grow them back. So to do that we use these bone anabolic drugs which basically mean they activate the bone to grow back. They activate those osteoblasts to put down more matrix and to regrow the bone that you lost. So like I said, these are being developed for osteoporosis treatment. I think they're in phase 3 trials right now. There's a few different companies that have these bone anabolic treatments and they are really new opportunity to build new bone for myeloma. So I'm talking all the time to these companies to try and say, give me some sample, let me try this. In my mice, let's do a clinical study. But I have to show them that it works in mice first, here. So so far this is, I know this is incredibly complicated. So I'm going to just try and make it as simple as possible. This is the basics, this is as hard as it gets in terms of the cell molecular level, molecular biology. So basically what happens is this little triangle, sclerostin, is a protein that blocks this other protein on the edge of a cell from binding to where it should. So for osteoblasts to be able to work, they need this thing called wint to bind to these different proteins that stick on the outside of the cell. But this blue evil diamond gets in the way and doesn't let this wint bind to where it's supposed to bind and then the bone can't grow and the osteoblast can't turn into a full functioning osteoblast. So what our job is, and we know that this sclerostin is induced and increased in myeloma patients and other people who have other bone diseases and it's because of these myeloma cells that there's all these blue triangles, blue diamonds. So what we try and do is block these. So the way we do that is we use an antibody which basically just is like a another protein that goes and binds and blocks sclerostin. So we call these anti-sclerostin antibodies. This is a drug, this is what they're treating osteoporosis patients with and as it binds them it basically sucks them out and it doesn't let them block wint. So then wint can bind with its receptors on the edge of this cell and this activates all these different signaling cascades and proteins and phosphorylations and downstream signaling molecules which lead to different genes and proteins being expressed and the most important thing is that this leads to something called osteogenesis which is new bone growth. So essentially if we can treat with these antibodies we can induce more osteogenesis and this has been shown a number of different times and the antibody is very successful so what we want to do is not just use this antibody for osteoporosis but I want to use it for myeloma and myeloma bone disease. Okay, so I'm just going to tell you what happened when we put it in the mice. So basically we wanted to see if these antibodies could prevent bone loss in myeloma and preserve bone strength and bone structure because we hypothesis that it can and we also have a second hypothesis that maybe not just, can we not just go back bone could it stop the tumor from growing. So there's a lot of evidence that the more bone you have the slower the tumor grows. This is because we think that these bone cells can actually stop the tumor cells. So basically what we're going to do is inject this drug into mice and then see if the bone can be regrown and also how the tumor changes. So there's a lot of different ways you can do that but the most common ways are testing in mouse. So we use different types of mouse models, we call them. So one is this complicated 5-2-GM1-GFP murine so that just means mouse. So it's basically you take a mouse cancer and you inject it into a mouse. The second way is to take a human. So a human patient who had myeloma we took their cells many years ago, grew it on a dish, we've been growing them and now we can inject these into mice and they grow in mice. We have to inject them into mice who have no immune system. So it's called a xenograft, like a species to species injection. And then we look at their bone, so trabecular and cortical changes which just mean different types of bone and then we do something called longitudinal BLI which is bioluminescence imaging which is a way to measure how fast the tumor grows. So it uses, because these cells that we inject from a human actually have a firefly gene that make them glow so we can use bioluminescence to see how fast inside the mouse how fast the tumor is growing. We can measure that over time. So then we have to do treatments so we looked exactly what they do for humans and we tried to replicate that in the mouse so we did a weekly anti-sclerosin antibody treatments or this control antibody treatment and we did weekly injection and we sacrificed the mice at day 28 and we look and see how the mouse is doing. So basically you can have naïve which is a mouse who has no cancer and then you have the tumor mice and then you compare the controls versus the drug and you see did it work, was it helpful or not. So luckily we actually saw when these are mouse trabecula of their femur and we actually saw that the mice who had tumor, the fact to GM1 actually had decreased amount of bone volume so they had less bone because they had more tumor but when we treated with the antibody we were able to see that these tumor mice when they had the drug actually had more bone so this is a good sign and there's all these other ways to quantify it this is basically an x-ray by the way so this is the spongy bone that you are asking about this is the trabecular bone so here's the edge, this is the cortical bone and this is the trabecular bone we look in the bone and we try and just measure how much bone is in there and then we can also look at the thickness of the trabecular so we have the software that goes through and it measures how thick all these little trabecular are and we also have something that measures the cortical thickness so it measures all of the cortical size basically and so this was also increased so all of the tumor bearing mice when these 5-TGM1 had the antibody it actually helped all of these properties to go up so that's a good sign Before you begin treatment or inject the proteins into these mice do you take x-rays of the bone prior to the analysis so that you can actually measure the bone loss or increase? In this, no, but we've done it before where you can see that there's bone loss but the problem is these x-rays actually have to be done on the mouse after he's sacrificed and dissected because we don't have this resolution unless we basically put this in a very small x-ray machine that is actually a micro-CT computed tomography machine so we don't have this kind of resolution so we can't do the exact same mouse but we can compare all between the different groups essentially and we did the same thing with the vertebra so we looked at the lumbar vertebra number 3 and we also saw that the naive control when he was treated with the anti-scorosin antibody has more bone when the naive control has tumor you can see that there's a huge amount of bone loss but that this is rescued when we have the tumor bearing mice that's injected with the antibody and this is quantified in the same way when you look at bone volume out of total volume it's just another measurement that basically says the tumor bearing mice that they do better when they're injected with the drug and then the thickness in the vertebra and also the cortical thickness so the software goes and computes the thickness of the cortex and so we wanted to see if this actually translated to strength so sometimes you might see more bone but you don't know if it's actually stronger bone so the way we test this is that we took lumbar vertebra number 4 and we put it in this compression testing device and we want to measure the peak load so basically how strong are the vertebra and luckily we see the same thing where we actually saw that this is when you compare here to here the tumor bearing mice have weaker bone the peak strength is lower but if you look at the tumor bearing mice who have the treatment compared to the tumor bearing mice who have PBS so nothing they actually are stronger as well and then we just do some significance assessments so I think this is another cool confirmation so what this assay is is basically a measurement of how much bone is formed so not just at the end with x-ray but actually it measures the kind of the length of the bone that's being formed so the green basically traces the osteoblasts so we inject two different dyes into the mice about a week apart and the dyes bind to the new bone so then you can measure how far apart the dyes are and you can go through and do a section on the bone and see if in the tibia if there's a big difference here that means a lot of bone formed in a week and if there's not a big difference between the two lines like here that means that not a lot of new bone is being formed and then we just have all these different parameters so we measure bone formation rate mineralizing surface and mineral opposition rate which all basically just show us in different ways that when these tumor-bearing mice are treated with the antibody that they have more bone that's being formed and so we can also do this in the other model so in the human xenograph model which means the mouse that was injected with human cells basically we see the same thing where the anti-sclerosin antibody was better than the control so increased bone in the femur in the trabecula of the tibia and also the vertebra so the second hypothesis was does this have any effect on the tumor and we got kind of mixed results in this so when we looked at the mouse who was injected with mouse cancer we didn't see a difference so if you compare these two groups on this graph and you do any statistics here you see no difference and when we also look at histology all the brown here is tumor so there's the naive control meaning he has no tumor here's the tumor-bearing mouse and here's the mouse with the treatment and at this time point at the very end there's no difference between them now one of the reasons might be that at the very end of this treatment all of the bone marrow is pretty much packed so if there was any difference in the rate of the tumor growth the slower growing one probably caught up with the control so this is just the end point when we sacrifice the mice but when we looked at the other model we looked at the MM1S model so this is the human mouse the human tumors injected into the mouse and in this we actually have the ability to track every week the amount of tumor that's in the mouse we do this with the bioluminescence imaging so we have these mice injected with tumor cells we put them into a black box and then we measure how much light comes out of the tumor cells and this tells us that the more light you see the more tumor they have so these controls these are the 10 mice in this group have a lot of tumor and when we compare it to the amount of tumor in the treated mice we actually see that these mice had a lot less tumor and sometimes it's hard to tell and so we have to use a software that basically recognizes how much light is coming out of each mouse so we draw a circle around each mouse and we measure how much tumor each mouse has and then we plot it here and we can see that actually the mice that are treated with the antibody do better than the mice that are treated with the control so for this experiment our conclusions are that the anti-sclerosin treatment prevented myeloma bone loss and increased bone formation increased trabecular and cortical thickness bone strength and suppressed tumor growth and at least in the ML-1S in the human model it suppressed tumor growth and increased bone parameters I have a question on the slide before when you have the two the control of four weeks and the anti-scleroma on the bottom it looks like there's one mouse that looks like he's fine why is it just that one it's true so we actually drop this mouse as well for the analysis and everybody wants to hope that it's a lucky mouse but the problem is he might have not been so to get all of these to have the exact same number of tumor at the beginning is tough and it could be that when we inject in their tail the tumor cells that not enough tumor cells went in to kind of engraft or grow in the mouse so basically we had to take him out for analysis just because it could have been user error basically if when you inject you don't get right in the vein and the tumor cells don't grow in the mouse you can't really be sure that the anti-scleroma is working so in the rest of our analysis we just dropped that but we still saw even for all the rest of the mice that even all the mice who have signal or have a tumor that they were all a lot less tumor than the other mice and that's why we have to do enough mice to really decrease the kind of error that you get between mice and the people who did these injections are actually professional tail vein injectors we pay them just this is basically their only job so it's a tough thing they're very small vessels in so basically what I'm trying to do now is talk to Amgen, Novartis and Lily and say look these anti-scleroma antibodies have potential to prevent bone loss increased bone strength potentially reduced tumor for my little patient so convincing them to work with me and allowing me to continue the studies is my current challenge so if they're listening I would love to keep working on this project but I'm going to just shift gears a little bit and tell you about one more cutting edge project that is going on right now in my lab like by my post doc and by my research associate like today as we speak and I think this is really exciting so something that we're learning about that we didn't even know five years ago is that in your bone there is fat in your bone marrow there are these little fat cells so everybody knows you have subcutaneous fat under your skin and you have visceral fat you have fat all over your body but most people don't realize that you actually have fat in your bone marrow and that it's not just stationary sitting there it's actually very active so it's producing all sorts of things that are signaling throughout to other cells all over your body so in high-fat diet for mice we give them a very fatty diet they get a lot of bone marrow fat and it looks like this but when we have low-fat diet they actually have less fat in their bone marrow so what we're trying to understand is what is this and I'm working with Cliff Rosen to understand what is bone marrow adipose bone marrow fat and how does it relate to different types of disease so we know that body mass index and abdominal fat is linked to your risk for myeloma it's not a strong risk but it's slightly elevated risk of developing myeloma for things like this and bone marrow adipose tissue also increases with age and with elevated BMI body mass index so it's possible that bone marrow adipose another cell in the micro-environment here in the bone marrow might be interacting with the tumor cell might be changing the metabolism of the tumor cell might be producing factors so we're trying to understand how these fat cells which basically look like this or adipocytes are interacting with myeloma cells and one thing that we're trying to do is make a 3D model of the bone marrow so the bone marrow is not 2D this is just one slice of the bone marrow but the bone marrow is a three-dimensional component and if we can model that in 3D the cells are very different and they act very different when they're in a 3D culture versus in 2D so in 2D culture our adipocytes get all these little tiny lipid droplets but in a 3D culture they become more unilocular meaning they have one lipid droplet which is what they're supposed to have so here it's just an adipocyte that has one big fatty droplet but when we grow our cells in a dish because they don't have three dimensions they can't pull onto things in 3D they're just squished onto a flat dish they're not able to form this three-dimensional cell and so the cells are very different they express different proteins and they act differently so what our lab is trying to do right now is develop a three-dimensional bone marrow adipose which means stop growing cells in 2D figure out how to make them grow in 3D so there's lots of ways to do that and I'll tell you about the way that we're trying to do this model is actually a physical a physical 3D model so instead of just a petri dish where the cells have to sit in 2D how can we give them the ability so right now they can't stack on top of each other if you grow cells they pretty much flatten out they have nothing to hold onto in your body they're in 3D because they have proteins and all these sorts of collagen that run along your body that allows them to all grow and that's why we're three-dimensional people and we're not two-dimensional people but in the dish we don't have that so we're trying to develop that exactly allow us a trellis some sort of sponge that's small enough that the cells can grow through that allows for fluid flow that allows for communication that doesn't cramp their style too much so before I show you the picture the way that we're that's okay the important thing about fat and the interesting thing about fat is that it actually comes from the same cells that make bone so your bone cells your osteoblasts and your fat cells all come from this one mesenchymal stem cell so when we're studying fat in my lab what we're trying to understand is can you change how these mesenchymal stem cells differentiate down this how do they choose which way to go they choose their fate they choose it based on signals you can put different things in the dish they choose their direction mechanical stimulation makes them more like into bone cells too much fat too much energy and often you can make them and other ways you can make them turn into fat cells adipocytes and this is true in the bone area and we're still trying to understand what are the differences that make a stem cell want to stay a stem cell or differentiate into a fat cell or differentiate into a bone cell and what we're thinking is that the more bone for my alone patients the better not only would it make their bones stronger it could potentially slow down the tumor but also we're still trying to understand what the bone marrow fat cells do so we're trying to understand do they produce lipids so give fat out that helps the tumor grow faster or other types of inflammatory cytokines we don't really know this but this is a really interesting cell type that now we're trying to study so the lattice that we're making is made of a protein because cells like to grow on proteins you can make it from collagen what we've chosen is to make it out of silk because silk is a protein that's very strong so when you're making an artificial environment if it's a gel or hydrogel often those are not strong enough and they're kind of like too hard for the cells to move through but if you can make like a spongy porous type structure the cells can hold on to it they can pull it so cells need to be able to pull cells can tell how hard they're pulling they can feel the forces of the the type of biomaterial that they're grown in so silk is an amazing biomaterial for tissue engineered bone so people are using this all over the world basically to try and regenerate bone and they put bone cells on it and they give it the right factors and they make bone grow out of silk scaffolds so this is actually in collaboration with Tufts University where I did my PhD in David Kaplan's lab so he and I are still working on this along with a number of other investigators so basically we get these silkworms we get them dead from Japan or Thailand and they come in these cocoons so as the silkworm spins up himself or herself in a cocoon they essentially come to us like this dead worms inside a cocoon and silk protein is spun out by the silkworm and so what we do is we cut the cocoon open and we take out the dried up worm and then we use that cocoon and the cocoon is made of two different proteins one is called sericin and one is called fibrin and this is the same silk that's in your silk tie or your silk blouse it's shown right here it's just a protein and you can dissolve this and make a solution and you can do basically anything you want this is just a protein solution so you can electrospin it into mats you can pour it into a film and what we do is we use dialysis cassettes get it to the right concentration of protein then we pour it into molds and then we pour salt onto these and then you have these little sponges which basically feel like kitchen sponges when they're soft and when they're wet but when you put bone cells on them the bone cells grow very well they attach to this sponge they grow in the pores they start to produce their own matrix and they make it hard and they make a mineralized 3D sponge and you can punch it big you can make it small depending on how big you want you can cut it as a square or a triangle whatever you want we try not to make them too big if they're too big they don't get enough oxygen on the inside so we keep them small enough so that we can get diffusion of oxygen and nutrients on the inside so these we call sponges or silk scaffolds and this is what it looks like with fat on it so this was some research that we just published with David Kaplan's lab about how do we use these to make white adipose tissue so we took adipose tissue white adipose tissue basically from lipo aspirate so somebody had liposuction down in Boston we took their liposuction we took all the cells out of it we ground it up and we put it on the scaffold to see what does it look like and there's all sorts of cells in here and a lot of them are these big red fat cells which are adipocytes and a lot of them are not fat cells they're the green cells that are kind of skinny and they don't hold any fat and those are just kind of these infiltrating fibroblasts and other types of cells so this is what it looks like and you can do all this kind of cool imaging to stain for different parts of the scaffold stain for different parts of the cell and if you're wondering what the purple is that's basically the scaffold and then all of these green and red things are all the cells the white adipose tissue in this three dimensional matrix and then you can do all sorts of 3D imaging and fly through with these special microscopes that we have that allow us to basically image these scaffolds so what our goal was in the last few months was to try and do this with bone marrow adipose tissue so a lot of people have done it with white adipose tissue and now we're trying to see can we recapitulate the bone marrow with the bone marrow fat cells and so what we did is we took bone marrow adipose well we took bone marrow stem cells and we put them on the scaffold and we put them in adipogenic media which basically tells the stem cell to become a fat cell and then it develops into these some of the cells not all of the cells these cells are not fat cells but these cells are and they start to get this big droplet of fat on the inside so hopefully we'll be able to use these and develop better models of bone marrow fat to understand these fat cells so I just have a last few pictures which are basically these are myeloma cells growing with the fat cells these are a whole bunch of non-fat cells but here's the adipocytes those are the fat cells and it's all in a 3D matrix the more you look at these the easier it is to understand but I know it's a little confusing to just see these but here's another representative example for you so this is the silk sponge this is the scaffold in purple and on the scaffold there's these cells that are growing and then part of the scaffold here is a three-dimensional mound of cells that's basically started on the scaffold and then produced its own matrix and allowed itself to kind of grow off and this is one of those slices and you can see that there's fat and there's nuclei which are basically inside the cell and then there's also these green circles which basically show you that that's the outside of the cell so this is the last picture which just shows that there's some fat cells and then we tried to put bone and fat together so here's some green bone cells and here's some fat cells that we put on top of the bone so I think this is the last summary slide and basically just that the microenvironment of the bone marrow is very complicated so what I just wanted to try and capture is that the myeloma cells here are growing and we know that they're interacting with osteoblasts we know that they're interacting with immune cells we know that they're interacting with osteoclasts those are the cells that eat up the bone and now we're starting to understand that not only are they interacting with these stem cells but we can change how these stem cells grow into bone cells or fat cells and we're trying to understand better how to do that and that might have different effects on how the myeloma grows and so if we can understand these marrow adipocytes, fat cells and understand how they affect the myeloma this might be a new target just like we have drugs we have a lot of drugs this phosphonates against osteoclasts we're starting to develop drugs against osteoblasts bone anabolic drugs like anti-sclerosis and antibodies but right now there are no drugs that target any part of the bone marrow adipocyte most people don't know what we should do with the bone marrow adipocyte we want more or less are there certain proteins that are coming from it we don't know but it definitely has a lot of potential as a new way to target myeloma and the last thing that we're looking at is how do other peripheral adipocytes so fat cells in other parts of your body what are they doing how are they contributing to myeloma so with that I just want to thank all of my collaborators like I said there's researchers at the Garvin Institute their Crouchers lab Novartis, McKayla Kniezel and Inna Kramer have provided us with the antibody that we tested Cliff Rosen and my lab including Heather Fairfield who's done a lot of work with us I still work with Harvard Medical School with Tufts University and basically we can't do it with all of our we can't do it without all of our funding that we work so hard to get that we will continue to work hard to get and the last picture this is our institute and feel free to come by anytime that you'd like or give me a call and I can also give you my email address so it would be great to show you the lab and show you around thank you it's right down the street on 81 Research Drive which is a mile less than a mile away you probably drove it's behind the MMC surgery center which is on Route 1 so go down Route 1 behind the surgery center it goes down a little bit it's basically all one building there's about 14 professors in there and we're all studying different diseases and different proteins and genes and my lab is focused on myeloma yeah this probably isn't up your study line but that's okay do you know if they're doing any research on why a cancer cell starts in the first place such as the yeah the yes that's a huge question yeah I'm sure they're totally right right they can feel what you're doing I want yeah if you start the cell in the first place yes definitely and a lot of research is put towards that so how does that first mutation occur and there's a lot of different reasons depending on the type of cancer some cancers we know some cancers we don't in myeloma for the most part we don't know mostly that's because there's so many different mutations that lead to things like Walden-Sromes and myeloma that it's hard to say it was caused by a mutation it's not a genetic thing we're starting to see that there's more familial inheritance than factors that we didn't know about before but for the most part myeloma is not driven by anything that we can really understand or predict and these mutations are pretty much just random and then if it's and most of your mutations cause no cancer most of your mutations cause cancer cause yourself to die but those certain mutations that occur in the plasma cell that actually stay and can cause the myeloma yeah people are still trying to understand how can we stop them but we don't have a great answer yeah yes just in general terms what is a blast? blast is sort of a cell so osteoblast is osteo means bone blast cell but often in terms of myeloma they'll talk about blasts being myeloma cells or blast count but it's it depends on what kind of cell just in my case I had leukemia AMF right and there was a lot of talk about the number of blasts early on so what would sound like there were bad things but what I'm pretty sure that in that case they're just referring to the tumor cells and I can double check and I'm happy to do that after but I'm pretty sure that when they say that it's usually just your tumor cell whichever tumor cell it is yeah at one point my doctor that even referred to my condition acute not my myelobelastic leukemia something like that yeah I think basically they're just saying that's the tumor cell yeah but I'll check on it you see they're focused quite a lot on multiple myeloma and myeloma but do you see that what you're doing that translates to the other with myeloma with myeloma yes so they don't grow in the same way and they don't have the same interactions with bone cells it's true but as we learn more about how myeloma progresses in the first place and how does it grow and that might tell us why does the myeloma induce this bone disease while other cancers might not in fact even in myeloma there's some patients who don't have bone disease and some who do and there's some stages where there's no bone disease and then quickly there will be bone disease and we're still trying to understand what is it and I think by understanding that better that might tell us more about switches that occur and transitions that occur and differences between the tumor cells so yeah I'm sorry about that I'm not really familiar with these blood diseases but I imagine that not only is it damaging the bone but it's also compromising the function of the bone marrow itself absolutely and so what are the what are the side effects of the compromised marrow yes so immune dysfunction is basically a huge problem so your marrow is where all of your B and T cells are made so if you don't have that your immune system can basically not be as strong as it should you might be more susceptible to other diseases if your immune system isn't as strong as it should be and if your red blood cells if you don't have as much if your red blood cell level decreases you might not be able to oxygenate your body the way you're supposed to so you know your system all of your organs need enough oxygen to be able to function so all of your organs would be inhibited and affected by decreases in in blood yes yeah when you have a cancer uh that mastasticizes to different parts of your body are those cells still the same that's a huge question the other part of the body that is where it originally started usually no and that is another tricky thing is that often we'll do sequencing and kind of compare the mutations in the primary tumor versus the secondary tumor and they some will have some of the same mutations some will be completely different and depending which metastasis site so we've done this in myeloma when you look at there's something called extra medullary disease which is when tumor basically grows outside of your bone marrow grows in your skin they can compare that to spleen metastasis to bone marrow metastasis in different types different bone marrow cavities and they can see that there's different mutations breast cancer often the primary tumor they'll go after the primary tumor because it has a certain mutation but then when they look a few years later at the metastasis it will have acquired these other mutations and it won't react to that first drug so often that's why the first drug maybe worked on the primary tumor but a few tumor cells got out and then they changed and they got new mutations and they're basically a different type of cancer so Does that mean that the cancer is it's squished over the past so it's what do you want to call it it's gotten used to the exactly yeah so it can evolve basically exactly what happens is that and even when so it can either be over time based on time or based on treatments they can kind of become resistant and then evolve they can go down different pathways and become basically different types of cancer so a lot of people are now starting to not just say we treat breast cancer with X and we treat prostate cancer with Y but they try and look at the individual clones and that's why we're trying like at Dana-Farber we're trying to basically plot it just like an evolutionary plot where they show monkeys become these type of monkeys and they show this type of clone this tumor became these two tumors that became these four tumors and they had all these different mutations and we're trying to understand how does that happen and how can we get the right drug regimen so this is why you can't just use one drug anymore usually they're trying to use different drugs that target the nation treatments that have attacked the cancer in lots of different ways but yeah the cancer evolution that's probably the biggest one of the biggest problems that people are trying to understand the biology of how that works but that's exactly what happens and that's definitely related to the stuff that we're doing so how come certain clones grow in the bone marrow and other clones don't and what is it how do these tumor cells evolve to become happy in their certain environment and the tumor cells get to the bone marrow they just sit there and they become senescent endormin and they don't do anything for years and patients have no sign of tumor but then something can change and the tumor can evolve and mutate and then they start to grow again so how does that happen in the bone marrow or other types of places so something that we're trying to look at too yep it's very tricky