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And one is the carbon monoxide actually protected this normal cells. And there's reasons for that. But even more important was that we could use 100 sometimes a thousand fold less of the chemotherapeutic. Hi, welcome to the science fair podcast. I'm your host, Susan Keatley. I'm a PhD chemist, writer, and I love talking to scientists. On the science fair podcast, I aim to bring you conversations with scientists doing fascinating, cutting edge work on all kinds of interesting phenomena, ranging from physics to chemistry to biology and even the nature of science itself. In this second season of the podcast, we'll start by asking each scientist a little bit about their journey to becoming a scientist. And then we'll talk about their research and how it relates to one or two of the high school science standards from that scientist state. So come along and tune in for some science fair. Our guest today is Dr. Leo Otterbine. He's a professor of surgery at Beth Israel Dekenas Medical Center and Harvard Medical School. Leo's research group focuses on the role of carbon monoxide as a therapeutic agent in medical applications, ranging from organ transplant to infection to cancer. Inhaled carbon monoxide is currently in numerous FDA trials based in large part due to the research in Leo's lab over the past decade. Leo is also chair of the Beth Israel Dekenas Medical Center Institutional Animal Care and Use Committee as the site minor for the BID MC Center for the integration of medicine and innovative technology and a member of the Boston Biomedical Innovations Center Technology Assessment and Development Group. Leo mentors and provides specialized expertise in entrepreneurial startup ventures for innovative technologies. Leo trains graduate students, post-actorial fellows, surgical residents and junior faculty in basic research, grant proposals and career guidance. Leo, welcome to the show. Good to be here Susan. It's a pleasure to be here with you today. I would love to start out by asking you to tell us a little bit about your path as a scientist. How did you get to where you are and were there any bumps or unexpected twists along the road? Well, with any journey, there's always twists and bumps. My career started. Probably I would say as early as middle school. If I could go back that far and say that was a career, which of course it wasn't at that time. My interest in science certainly started then. And for the years, I chased it. I was always interested in biology. I was always interested in how science led me to questions about what was going on around me, whether it was my own body's reactions to things or whether it was things I would see in the environment. It always struck me as just an intriguing area to study. And that evolved as I moved through high school and into college. And I had visions of grandeur that I was going to be a medical doctor. That didn't, that my path changed there. I was not good destined for medicine. As it turns out, I was destined for research and science of discovery, if you will. And so that progressed as I moved through college into career post college, where I started at a biotech company. And that was an interesting transition for me to move from discovery science to biotech drug development. And then it sort of struck me that I needed to make more of a, or have more of an impact. So I decided to go back and get my PhD in physiology of all things. And so my road traveled multiple states, multiple laboratories, multiple experiences. I'm honored by what you described as your introduction of where I've been and what I've done. And I'm proud of all that, of course. And I look forward now to mentoring the next generation of scientists and share more of what I've discovered in what our research has found out. So I'm just going to, this could be your. That's great. Thank you. So carbon monoxide is a, a focus of your research and it has been for a long time. Most people think poison when they hear the words carbon monoxide. But in your work, carbon monoxide is anything but a poison. So can we start by having you tell us a little bit more about what carbon monoxide is and what were the early observations that hinted this might be something more than a poison. There might be something interesting here that is unexpected. Yeah, so I'll start off by saying that everything is a poison. That was coined by a very famous Greek man named Paracelsus who said everything at the right amount is, is dangerous and that includes oxygen and water. And carbon monoxide would fall into that bin as well. You know, carbon monoxide has been around forever since they figured out how to combust wood into fires. People knew that it was present and it was some of the great scientists of our age that used it and understood it as how it affected the body. And they used it actually as a tool to understand how blood carries oxygen. So it's, it's spent the last 150 years or so under the under a under utilized mostly as a tool if I would say and it was not until the early 1990s where maybe even predated by a couple decades there, probably the late 1960s, I would say where they realized scientists, very, very wise scientists, realized that all the cells in our body generate carbon monoxide all the time every day. So much you can calculate how much is generated in the body of a healthy individual. And over the course of the following decades, they realized that it had biological effects. But no one really accepted it as a molecule that would have anything but toxicology because and also occurring in the 1960s and 1970s was this idea of pollution and that one of the pollutants that was most dangerous was carbon monoxide. And there's no doubt that if you have enough carbon monoxide, much like anything else, it will do things to the body in ways that are coined as toxicology or toxicological effects. And I think the media and the literature and those who wanted to promote carbon monoxide based primarily on pollution, I would say, wanted it to be a toxic gas and something to avoid. And if you look at what present in exhaust or even burning fires, there's over 500 things that are present in that exhaust or those burning fires that are dangerous. And carbon monoxide remains the one that people talk about and in my early sort of career as a PhD student, we were asking the question, why would the body generate it if it was dangerous? And that's sort of where it sort of took off as far as my research was concerned. Yeah. And can you tell us what were some of the early ideas? Why would the body generate it? So there's an enzyme system for those who are interested in enzymes. There's proteins in the cell and there are those enzymes that are necessary for degrading things, breaking things down. And one of those enzymes that became interest, that of interest to me was one called hemoxygenase. And hemoxygenase was discovered in 1960s, 1968 to be precise. And its sole role is to break down molecule called heme. And that's present in, the heme is present all throughout the body. It's a critical component of red blood cells, as you may know. And there needs to be a process to remove it. And as it's removed, it's broken down. And one of the things that's released, one of the molecules that's released is carbon monoxide. So we knew not only us, but others that if you have high levels of this hemoxygenase around, it was coined and actually labeled as a protective gene. And no one understood why. And for decades, most attributed it to the fact that its end product of its reactions was something called Billy Rubin, which is a yellow pigment. And Billy Rubin is a very powerful antioxidant. And so everyone said that there must be the reason why this hemoxygenase is beneficial or helpful. And it was sort of a question that I ask as a PhD student. Maybe there's something to this carbon monoxide that's produced that could also have and provide an explanation as to why this enzyme system is beneficial. And that's sort of what started the work in the late '90s for me. And it's progressed for the last 25 years. Wow. And what is the relationship between the carbon monoxide and the body and inflammation? Good question. So it turns out that if you were to measure carbon monoxide in the blood or if you could measure it in the tissues, it's elevated in instances of inflammation. And this spans which we knew from work of ourselves, of our own, as well as others, that the disease progressed, whether it was infection, cancer, diabetes, organ transplant, which I think we're going to touch upon. But the levels of carbon monoxide in the blood actually rise. And so it begs the question, why would the body produce a molecule, in this case, carbon monoxide, in greater quantities when the body is otherwise carbon monoxide? compromised and is already suffering and battling a disease or some sort of Melody that needed to be dealt with why would it create a gas that would be further detrimental and that's really what would prompt it Some of the questions I asked in the early days Well, and I think we will use that to get into where I'd love to Spend maybe the majority of the time this morning, which is the story of Carbon monoxide and organ transplantation You know the the many twists and turns on Discovering its role and then where we are today Yep, so I was fortunate in my travels. He talked about bumps and and Things that I had to overcome and one of those was moving around the United States And it might work the road took me to the University of Pittsburgh Medical Center Where I was just beginning my research into carbon monoxide how it was worked and that working and I was a junior faculty So I was trying to get my career started writing grants and things like that and I was fortunate to be just by chance in a laboratory that that was next to all the surgical All the surgical departments labs and one of those primary areas of interest was organ transplantation Just by chance one of those one of those fate things you could talk about but I found myself Interacting with with fellows and residents and junior faculty about what my work was normal conversations And I knew at that point I had published work that said that the carbon monoxide was anti-inflammatory And we have reasons to believe how it was happening and this intrigued these organ transplant researchers and the very famous Starsal Institute is based in the University of Pittsburgh and it trains Surgeons from all over the world and they have gigantic facilities and so it became very easy for them to use my knowledge and my apparatus that I developed as my PhD part of my PhD work To allow them to study organ transplantation because it involves inflammation primarily and It it rocket shipped out of that's the right term Rocket it as these as these surgeons practice their techniques using animal models of organ transplantation and they transplanted everything they could heart lungs kidneys liver Small bowel all these became part of their Research and why not throw water binds carbon monoxide in there because it'd be simple enough to study and if it has an effect on inflammation then this would be a good hypothesis to test the carbon monoxide would be beneficial to the organ with transplantation I was fortunate at the same time which preceded a little bit of this to be connected again one of my bumps as I had or my Fate if you will I was fortunate to meet a group of immunologists at here at Harvard Medical School And they were also interested in in carbon monoxide and primarily the hemoxygenase story and I Was fortunate to be able to collaborate with them and they also were transplanting not just kidneys from this animal to that animal But they were doing Xenotransplantation and I'd probably say they were the first to to very to initially show the benefits of when you transplanted a Mouse heart into a rat body and I can talk about that if there's interest That carbon monoxide would prevent the rejection of that of that That heart and it was primarily driven by the Anti-rejection capabilities of the anti-inflammatory effects of the carbon monoxide to protect that organ and or Limit the host response so that the recipient of that organ usually mounts it a response that rejects the organ Mm-hmm. It just it just calmed if you will the the immune response such that the organ would survive and so combined with what happened here at Harvard and what was really going on and And spearheaded by a lot of the group a lot of the groups that surgeons at Pittsburgh the organ transplantation field exploded with carbon monoxide So much so that it garnered a lot of attention by those outside of academia and Lured in or brought in those who were interested in gases as their business model and this this brought the Europeans it brought the US Gas distributors those who provide oxygen to hospitals those who provide other medical gases nitric oxide is another one to have an interest in developing carbon monoxide and Sheerly because of the volume of work that occurred with organ transplant and by this point it It started to make its way through the large animal models so they were doing now pig transplants and my lab was Was provided the resources to test a delivery system the first delivery system to deliver carbon monoxide to a in this case a pig But designed for a human that ultimately led to the clinical trials that were done with kidney transplants So we we were the first to perform the the testing of a very innovative device to deliver the carbon monoxide during a surgical procedure in the operating room We tested it and validated it in pigs and the carbon monoxide showed benefit in an organ and a kidney transplant model And meanwhile all the all the all the organ transplant Researchers in the world were continuing to study why it was working and how it was working and where else would it work and There became a very big interest in would the would it be best to treat the donor of the organ would it be best to treat the organ? just self or the recipient or all three and that question we still remains In terms of the organ transplant field as a whole Leo to that point if you could back up a little in those early tests were you treating the recipient and I would love for you to give specifics on like exactly how effective the carbon monoxide was like in terms of you know How much longer the animals survived? et cetera after the transplant So I'll so I'll use the story that happened here at Harvard because I think that was probably the earliest and so I was Continuing to finish my PhD I happen to be at Yale that at that point another one of my moves and I was contacted Our lab was contacted lab I was working in by the group here at Harvard and they said we've we've been studying this hemox The gene as enzyme and we've seen some of the in time flammatory work you're doing would you mind meeting with us and We came up to Harvard to the behavior to the Bethes rule and they had a model of zeno transplantation Which I mentioned where they were transplanting mouse heart into rats and so what they wanted to test was would it be effective and So I brought my apparatus up here They transplanted mouse hearts into rats and in that scenario it because it's a zeno transplantation There's what's called hyperacute rejection and those mouse hearts in a normal animal will fail They'll stop beating with in days and sometimes with an hours transplant and What they wanted to do which was based on input for myself was if we put this animal into the chamber into the exposure system Which I developed and in this case we're treating the recipient Right, okay, the rat was being treated so after the heart had been transplanted it was put into the rat Okay, I mean what the rat was put into the chamber and the chamber the rat is breathing in carbon monoxide Correct, so we we were only working with inhaled versions of carbon monoxide which stems from a lot of a lot of background Research to determine how much to give and when to give it And we were giving very low amounts think the point here is that we were giving very low Amounts that were not detrimental to the animal in fact we did a whole series of studies to demonstrate that And so the I left the taught these individuals people here how to do the exposures and I left and went back to my work And every day I would get an update about how the How these animals were doing and the way they test this in in this particular model is the heart is not Transplanted into the chest doesn't replace the rat heart what they do is is is a very Very very fine surgical technique to transplant the heart and connect the vessels of the heart into the abdomen of the rat Wow, which requires very precise surgical technique and So much so that the sutra they use you can't even see it without a microscope How they do this is just amazing to me, but the heart will will take blood and it will be Continuously until it's rejected and the way they test that is they just hold their hand against the abdomen of the animal And they can feel the heartbeat of the mouse in the abdomen Wow A very crude way to know if the animal if the heart still surviving and so every day they would measure it and every day They would communicate back to me the heart still beating and they got to that critical window if I don't remember off the top My head if it was two days or three days and three days it's still beating four days it's still beating five days seven days Ten days it's still beating and I got exclamation points in the emails after it's still beating in 50 exclamation points They couldn't believe that it was going to work the way it did and that really that really A tract of a lot of the team Was was the rat in the chamber this entire time like over those ten days or would the rat be kind of taken in and out of the chamber? Yes, good question the in the design that we originally it was changed over the years But the design we originally settled on was one hour per day so the animal would go over an hour and and And breathe the carbon monoxide and then spend the rest of the day not in the chamber So there were different variations of that but that was more or less that we used that feels like a very small dose It was and so the idea was that for from a biological scenario. We were keeping the inflammation at base at the low level, so that there wouldn't be an immune response to reject the organ. - Right. So then you were saying, this got a lot of attention, this experiment. - This is really what spearheaded a lot of the work that continued at Pittsburgh, where they did a lot more organs and a lot more in-depth sort of studies and attracted the companies that wanted to develop this as a product. And it proceeded that way for a couple more years where they developed this device, we tested it in pigs, and that really was what drove the clinical trials. And there was a clinical trial that lasted a couple of years back in 2008, I believe it was when it was going. And the idea was to treat, in this case, the recipients, so those who were gonna receive the organ, and that's how we studied it in pigs. - Right. - And they were only gonna be getting the carbon monoxide during surgery. So at the anesthesiologist was in control of the gas, and he would turn it on when the surgeon had a precise time when they said, okay, turn it on. And that during the surgery, the patient was breathing carbon monoxide. And then when the surgical portion was complete, the surgeon would say, okay, I'm complete, and the anesthesiologist would turn off the gas, and the patient would be brought into recovery, and their kidney function, 'cause it was a kidney transplant, was monitored. - Right. And these were, these were in humans, receiving kidneys. - In humans? - Yeah. - There's 28 or 30 individuals on this planet that are walking around, hopefully still. This was 15 years ago, that have a kidney, that they received, and they also were given carbon monoxide to breathe. And the important to this is that the thought was, what you started up this whole session on, was carbon monoxide viewed as a poison. So my take on this was that when those individuals were asked, consented, would you participate in a trial where you're gonna be delivered, or you're gonna be given carbon monoxide, that most would say, are you out of your mind? - Right. - I'm having a kidney transplant. Why would I wanna be carbon monoxide? I don't know about that. And many, most accepted that, that would be a good thing to do. And so to their credit, they took that gamble, and they, their kidneys, which when they were looked at as post-surgical, did better than those who didn't receive the gas. They inhaled, see. - Wow, wow. - So they just, so the sad part about this, that part of the story, another bump in the road was that the company decided as companies do, we're not interested in this as a business anymore. We're gonna get out of the gas delivery business, and we're gonna use more traditional types of drugs that you take by mouth. So thanks for all your help, but we're gonna close this down. And that's sadly, what happened with the inhaled gas route of things, which was a very big disappointment, of course, but the field progressed and still is going today. So there's been ups and downs. - Right. Why do you think people were so open to try and carbon monoxide in this trial? - I have to believe that those who present the idea, and this is mostly surgeons, anesthesiologists, others, that you really have to convince somebody that there's a reason to do it. And I think that that was very well captured by those who were presenting this to a kidney transplant recipient that your organ is gonna do okay, we hope, after surgery. But there's a high likelihood that if you breathe carbon monoxide, it actually could do better and your hospital stay might be shorter, and the need for something called dialysis might be shorter. And there may be even the possibility that the rejection, the anti-rejection drugs that you are gonna have to take for years, if not your entire life, may not be necessary for the longterm. If your kidney starts to function more efficiently. And so I think combined with a sales pitch for lack of a better description, compelling argument made by what the data, and all the data is presented. Now this was done, Mr. and Mrs. Jones and pigs and showed this tremendous benefit. We think this is good for you, was probably a big part of that. That was something that people believed was a good thing. And it's amazing to me still. - Right. Are there ongoing trials now? - So the field evolved, I don't know how long, how much more you wanna know about the carbon dioxide field in terms of where it's gone, but the field evolved. - Yeah. - It moved away from the inhaled route, which there's reasons for that. And now there's companies that have developed drugs that are standard, they're typical, I would say like a pill, who would take my mouth. There is a company in California that is developing and is now in clinical trials with a carbon monoxide drink. Perhaps the most intriguing one is of the most recent. There's also a company in New Jersey, I'm just to give a feel, just to cover the basis, that is giving carbon dioxide into the blood stream. They give it as, they expose the blood to carbon monoxide, then they give the blood back to the patient. - Wow. - So that would be cool. And then the more recent one, which is the more intriguing of late is a group at MIT, developed their bioengineering department, group over there, developed a carbon monoxide foam, or what they call a hydrogell. And even a solid form, much like the candy pop rocks, where you can believe carbon monoxide, instantly as if you're drinking, it's the same ingredients, the exact same ingredients that are used, you put on top of our cappuccino or a caffé latte, it's the same ingredients, except instead of using air to make the whip that you put on your caffé, they call it molecular gastronomy, because they convert products by just infusing it with carbon monoxide to make it into a foam, or a gel, a cream, a solid, just about anything you can imagine. And that's now being developed for treatment of different uses. All of those that I've just mentioned are being tested in different areas, not a organ transplantation, but there's reasons for that, but they're all moving along in their trials, which is good. And hopefully we'll see the benefits of carbon monoxide somewhere, somehow, the one area that's probably the furthest ahead, if I had to pick one, would say that carbon monoxide being used for sickle cell anemia, that's actually suffer from tremendous pain, and it turns out that that's a benefit. And those with Parkinson's disease, where the individuals funded, and have interest by Michael J. Fox's foundation, the ability to slow the symptoms of Parkinson's. So there's really quite a span of disease indications where carbon monoxide has shown to be beneficial in addition to organ transplant. - That's so interesting. And all you think, or people think, really has to do with minimizing inflammation. - That's at the core. It's funny, I was just at a meeting where someone asked me that same question. And if you'd asked me that question 20 years ago, I said, yeah, it's all inflammation, but there's more to it now. And I think some of what we've talked about, I think is in some of the areas of interest for this call, is where does carbon monoxide act? And how does it act? - Right, right. - And maybe that's a transition for us here, is to say that we now know much, much more about how carbon monoxide changes the behavior of a cell. And if you're talking about an immune response, that's inflammation. And the carbon monoxide changes the immune response. If you're talking about something that doesn't necessarily involve, and it's hard to separate, inflammation is such a part of every disease path, whether it's Parkinson's or sickle cell, or even organ transplant, separate out what's inflammation and what's not inflammation, but there are other cells that are not immune cells that become the target for carbon monoxide. Probably the easiest one to understand would be a cancer cell, where a cancer cell is not an immune cell, it's not inflammation per se, a tumor is inflammatory, but a cancer cell is not. And if you give a cancer cell carbon monoxide, it stops growing. And the reason for that is because cancer cells require a certain type of metabolism in order to continue, and this carbon monoxide interferes with that. - Wow. - And it's ability to act to function at a bio, from a bioenergetics standpoint, if that's makes any sense. It changes its ability to survive as it normally would. So that's not an inflammation per se, but it's definitely a cellular process that is changed when the cell sees the carbon monoxide. And in the case of cancer, it would differentially stop cancer cells, but not surrounding cells that are benign and normal. - So one of the studies we did, and has now been done by many labs, is if you give normal cells carbon monoxide, they actually do fine. You wouldn't know they were being exposed to carbon dioxide. So subtle changes that are more molecular, that we could go into, but probably outside this call, but the idea was that the cancer cells, not only would this carbon monoxide affect the growth of the cancer cells, in terms of their proliferation, their doubling of in their cell division, but if you throw a little chemotherapy on it, which is one of the questions we ask in my lab always is, what's the relevant of this to the clinic? We're not interested in necessarily in curing diseases and animals, although that's one of our side benefits, is that there are veterinary applications for all this, of course. But we're interested in how this might translate back to a human disease. And so when we model this in animals, we try and do it as similarly as possible to human. And so we add chemotherapy to a cancer study, right? So we add the drugs that would be used in normal cancer treatment. And we're interested in, What we find, which is really quite fascinating, is that the problem with chemotherapy is many know, is that there's side effects. The side effects are things like the kills the bone marrow, the normal cells of the body. You sacrifice that to get at the cancer. The chemotherapy affects the growth of the cancer, but it also affects the growth and behavior of normal cells. What we found was that two things happened. One is, the carbon monoxide actually protected this normal cells, and there's reasons for that. But even more important was that we could use a hundred or sometimes a thousandfold less of the chemotherapy. Then we needed to do it without the carbon monoxide. So what that's been said, and in terms of chemosparing, is that that's suggested that we could eliminate the side effects, or many of the side effects, by just being able to use a lower amount, but still have the benefit that the chemotherapy had on the cancer. So you're not going to go into a cancer trial and say, we're going to not treat you with normal treatments, because there's many, many drugs for cancer. Yeah. You're going to have to have that anyway. Right. So we just want to add carbon monoxide. If you can protect the normal cells, lower the amount of chemotherapy, it was remarkable to us that we could see that. That's really incredible. I mean, just to speculate, how far out do you think that would be for patients? In terms of someone being able to get a cancer patient, being able to get that when they go in for treatment? So we've got to have somebody, one of those who are developing this as a drug, chase cancer, cancer is always a tough one to chase. And so once that, once someone becomes interested in that, it would certainly be, I think, a very quick translation, because you can't really, there's not much more you would do at this point. There are things like compassion trials and where there's no other option. So let's throw in something that's new and innovative. That's where carbon monoxide might fit in to a normal standard chemotherapy regimen. There are cancers that are just much worse than others, brain, for instance, or pancreas. Those are possible places where carbon monoxide would be tested first. But sadly, or not sadly, unfortunately, you've got to have someone who can drive those kinds of trials. And those are challenging for lots of reasons, one of which they take a long time. And there's a lot of competition. And so you're up against things that are not scientific per se, but more business related to my opinion. Right. Right. I want to follow up on something you had said, you know, in addition to minimizing the side effects of chemo, the carbon monoxide protects the normal cells. How does it do that? So the chemotherapy, in this case, damages those cells that are replicating typically. Yeah. It has effects that span, depending on the chemotherapy, they damage the cell just because they're toxic, right? So the idea is that you give a medication or a medicine, in this case, a chemotherapy that's targeting fast growing cells, and you just want to kill those fast growing cells. And the chemotherapy is worked by things like damaging DNA, damaging cell membranes. There's many modes of action of these chemotherapeutics. And so you're sacrificing some normal cells to hopefully get at the, at the, not, you know, the cancerous cells. The nice thing about, and the good thing about, and why I love biology is that the cells designed very elegant means to repair it. So if you treat a cell with chemotherapy, many of those, all the cells in the body are going to see the chemotherapy if you deliver it intravenously, for instance. And so many of those, you know, all the, all the cells don't die at once. You still survive the chemotherapy. So it's a portion of them. And that's because the cells have this very elegant way of repairing themselves. There are DNA repair enzymes that exist that if the chemotherapy comes in and it damages that DNA, as it says, how it works. The cells is, oh, we have a break. We need to fix it. And it just goes about like a mechanic and fixes the DNA. Cancer cells don't necessarily have that capability very, very well. And I'm not a cancer expert to give you all the details. But that's taken advantage of by those who are developing chemotherapeutics. And so there's, there's known effects of these drugs. And carbon dioxide goes in, we think, in many cases, and helps that repair process. Wow. It's their resistance to the damage and helps the cell survive. And essentially repair was done. What's injured or damaged? Very interesting. Coming back to organ transplants, what did you learn in that process about trials, clinical trials, and you know, moving something from the lab, really translating it to then something that patients can use? I learned that it's a very arduous process that takes, and the biggest enemy is time. Obviously, you need a lot of money, a lot of life resources, and those who are experts in so many areas of drug development. And that was some, a place where I learned an enormous amount about how a drug proceeds from an idea to lab through trials, even through to meet with the FDA and with the FDA and that's a very important component of the carbon monoxide because the FDA was perhaps the biggest challenge. I won't use the word enemy, but biggest challenge because they, like everyone else in the world, was stuck that carbon dioxide is poisoned. Why would you want to develop it as a treatment? And speaking specifically to carbon monoxide was one of the biggest, if not the biggest hurdle. And so those who were interested in it needed to devote that much more time, that much more money to showing improving that the amount of carbon monoxide they wanted to use for the trials was not as toxic as most would seem. And they insisted, the FDA insisted that the brain was the place where carbon monoxide was going to have the most toxicity. So the company did its diligence and its rigorous design and said, we're going to prove that there's no adverse events, they call them, to the brain, with unhealthy people. But that's always how it progresses. You start with, does carbon monoxide have a negative effect on a healthy person and then move into the injured of the disease? And that all progressed cleanly with carbon monoxide. But time is a big one. Organ transplantation is a challenge, of course, because you have this idea of treating donors, the recipients or the organ. How do you work out the logistics? If you treat a, someone who's going to be an organ donor and you're interested in the kidneys, if you're treating them with carbon monoxide, it's going to affect all, it's going to go to all the organs in the body. Right. So that question came up. No, what happens to the other organs that are not part of the trial, but still can be used. And would we have to have consent at multiple hospitals because the organs go to different places? Oh, that's a great point. And then also what would be even be your time scale for knowing what the effect are on all these organs? And you had to have all the training in place if you're going to treat, say, a remote hospital in Western Maryland, for instance, are they going to have to be trained on how to treat with carbon monoxide if they're going to treat the donors? So the recipients became the obvious place to start because that can be more controlled. All the organs, the only organs of interest were coming into that place for that surgery. And that became the primary. But when you talk about organ transplantation and things like how far can you transport an organ? Yeah. And it's a very small radius for many organs. But if you could extend that to say the heart and the lungs are usually the hardest to transplant, meaning you have to transplant them very quickly. So the radius of area of where you can transplant that organ if it's in Western Maryland, it's not going to go to California. But it might be the best fit a person in California, but it never will make it that far. So there was this plan that you could put carbon monoxide in the solution that was organ was shipped in and that that would keep the organ healthy enough to make the trip from I'm using Western Maryland because you're in Maryland. And I grew up in to California and be able to fit a recipient that was a much better match. So the other one other thing with organ transplant that's a challenge is that the organs come from many different sizes, shapes of donors. And some of them are some of them are donors or individuals who may be say in their later ages, I'll use that word, they're wise ones that will say that they have other maladies like diabetes or they may have arthritis and they're taking different medications. And those organs become a challenge because if that individual sadly is in a position where they can donate their organs. But the organs are coming from a body that is not 25 years old and a bodybuilder and healthy diet in perfect condition. You say, well, if it's a 60 year old heart from a diabetic, do I want to transplant that organ? And there's many, many surgeons that will say, I'm not going to take that chance because the likelihood of that organ failing is high. But if you could tell me, Otterbine, that the giving the carbon monoxide would help that organ, I'll transplant it. Because I have a better feel that that organ is going to do better because you're giving it a better chance by giving a carbon monoxide. So there was this enormous eruption by the organ percurement organizations to say, how else could we apply this? And it was cool. It was really interesting to see how that progressed. I feel so hopeful about carbon monoxide. I mean, from these stories with organs, but also all of the other conditions that it's shown to be useful in. And I think also just this general concept of it seems to be reducing inflammation, helping and repair, all of that. What-- you know, if you were to just flash forward 15 years, what would be your vivid hopes for how carbon monoxide would be used? I think the-- I think the-- that's a great question. At this point, I'm just hopeful that some group, some company, some motivated individuals will move this into something, given the breadth of where it's shown benefits. It's really-- I'd be happy with seeing it anywhere, be honest. I think the-- if I had to pick, it's unfortunate that organ transplantation isn't at the top of the list anymore, simply because the volume of data that supports that is so strong. None of the efforts right now are chasing that, and there's reasons for that. The-- there is a concerted effort right now to use this topical-- I mentioned hydrogels and films. And so the military, the Department of Defense, is interested in application of this to their-- to a wounded warrior, wounded war fighter, where you might be able to ascend this into a medic-- ascend it along with a medic that might be able to apply this-- Yeah. --in the iter as they say, where there's a active soldier who's been wounded and can't be removed from the-- the primary site, the role of care. And so that might be a-- sort of a Department of Defense military application, which would be very exciting to see that this would help support those who are helping defend us. Absolutely. And to me, that's a patriotic sort of application, if I don't sound too patriotic. But I think that's a really a util-- a place where it would be heavily utilized and really beneficial. The trials with Parkinson's, of course, would be very extremely impactful. It's anemia worldwide. This is not just the United States, but Central America, South America, Africa. The areas where sickle cell is so prominent, this would be such a help, potentially, for them. 10 years, I'd say that the likelihood of it being in one of those is high right now. And it's, for lack of a better word, a race to see who's going to get in their-- Right. Right. Right. That's super exciting. I feel like every now and then a drug or a molecule comes along that just seems to have all of these wide-ranging benefits. And of course, I'm thinking right now of these GLP agonists, aka, EZMPIC. But we've seen they help with glucose management, with weight, with potentially cardiac benefits, with benefits, possibly with addiction. There's something bigger at work. And I just am getting that feeling with carbon monoxide too, like these applications are just so wide-ranging. So it's exciting. Yeah, I think that stems from my belief. And nothing's ever-- drug development is such a tough road to travel, because there's so many places where it could have issues. The FDA, for instance, is its own battle. But yeah, the fact that the body generates it naturally, and the question that comes up that should come up, is if the body generates it, why does it need more? And it's a valid biological scientific question. And the answer I give is we have ways to take the ability of the body, the animal, in this case, or the cell, to take away its ability to generate carbon monoxide. And when you do that, when you take away the ability of the body to generate carbon monoxide, you have a very bad situation on your hands. The animals do terrible. They are much more sensitive to any kind of disease challenge, whether it's effective cancer, or you name it. They have a remarkable inability to battle against those things. So our working theory, our working hypothesis is that the body can mount a defense to a point. And then at some point, it maxes out, if you will. And that's when supplementing with something would help the body a little bit more. So a disease that you and I are-- if you get a cut on your hand from something, it heals. But if you don't have the carbon monoxide necessary to generate-- and that requires the system in order to heal properly. But if you have a major gash along your arm from something more harmful, the system just can't manage that as easily. So that's where supplemental might, we think would be beneficial. Yeah. Well, I'm just waiting for my carbon monoxide pop rocks. Maybe the supplement market will approve them first, because I feel like that's an easier place to get things approved sometimes. I would love to transition to some high school science related questions. OK. So I looked at the Massachusetts state high school science standards. And there's one in the life sciences that states-- students should be able to use a model to illustrate that aerobic cellular resp-- excuse me-- aerobic cellular respiration is a chemical process where the bonds of food molecules and oxygen molecules are broken in new bonds form, resulting in new compounds and a net transfer of energy. And then there are these various supporting statements. And one of them said the model should include the role of ATP for energy transfer in this process. And I believe that when we spoke before, we talked a little bit about the mitochondria producing energy and then it stored as ATP. And I think we had talked about how carbon monoxide was playing a part in the system. So can we go there? Sure. So you're dating me on my biochemistry, but I'll do my best. So the idea is that, as I mentioned, I think earlier that maybe I didn't. There's a mechanism-- I've broken it down into two things. One is if you say, what is the mechanism of carbon monoxide? I'd say something like inflammation. If I said, what was the molecular target of carbon monoxide? I'd have a different answer. And the answer there would be carbon monoxide likes to go where oxygen goes. And so that target becomes a molecule called heme. And you might link this back to what heme oxygenase does, which is it breaks the heme into pieces. So carbon monoxide and oxygen and other gases like to go where there's this heme. And the heme is part of the enzymes. And one of the biggest places for heme to hang out to use scientific targets is in the mitochondria. And the mitochondria contain many enzymes, proteins, that contain heme. And they use that to help generate oxygen and ATP. And so the ATP that's generated is necessary for life. And so one of the challenges to carbon monoxide work as a whole was that most would say, because it goes to the mitochondria, it's going to prevent ATP from being generated. And that's why it's detrimental to the cell and to the body. Because it prevents the need of the cell to continuously generate ATP. And if you can't generate ATP, you're poisoning it, which is why carbon monoxide is one of the reasons it's labeled as a poison. And what we found was that when we treat-- so we went into it thinking that, OK, well, that might be a reason why carbon monoxide is beneficial. We took the other way and said maybe it by changing the mitochondria ability to generate ATP, maybe that was a way for it to help the cell. This still will up for discussion. But I can tell you that research by us and others have shown that if you treat a cell with carbon monoxide, that the amount of ATP actually increases. Wow. Instead of looking at this, that carbon monoxide is bad because it prevents these enzymes from working that contain them, it actually can help them function more efficiently and generate more ATP. Wow. And so in terms of cellular bioenergetics, it seems that based on what we know, that carbon monoxide actually can help generate more ATP. One thing I'll add to that, which maybe fits the conversation here, is that there's a general theme, I'll say, of something called reactive oxygen species. And you hear about antioxidants, vitamin E, and things like that. And so these are touted as being dangerous to cells. If you have obstinate stress, then you're producing what are called radicals, which are free electrons, that damage because they're reactive. They damage membranes and proteins in DNA. And so there's something to avoid. And so the question was, does carbon monoxide have an effect on react these reactive oxygen species? And it turns out that you need reactive oxygen species, like hydrogen peroxide, superoxide anion. There's different ones. And they actually help the cell communicate within itself and among themselves. So intercellular intracellular. communication and you need those radicals and those arise in large part from the mitochondria. Wow. Getting into more of the biochemical biochemistry if you will. Yeah. The cord monoxide seems to have a dual effect in them in the cell at this level of the mitochondria. There's other ports of the cell where it acts which we can talk about but mitochondria is a big one. Yeah. And ATP and radicals are part of all the effects that carbon dioxide has. It needs those. It helps with those. That's really neat. Were you also saying when we spoke before that the mitochondria shape mitochondria and shape is not what we usually you know there's like a picture in the textbook and it always looks like a bean but it's like doesn't really look like that. Can you can you share that? I think that's really important. I think it's very important and it you know after 40 years of biology under my belt I went to a conference at the NIH about mitochondria and they were talking and now with advancing imaging techniques and things like this they were able to show what a mitochondria really looks like and so from textbooks as you know all too well that they look like little beans right like a fava bean or a kidney bean if you will and it turns out that that's just if you caught if you cross section through what mitochondria really are which are these long strands of like spaghetti that are intertwined and look like essentially a ball of spaghetti on a plate and they're not this bean shaped organism that we all think they should be look look like they're not at all so it's amazing to me when I saw the pictures these molecular imaging pictures of what they really look like it's wow change my view of them obviously completely that's great and I'm gonna look some of those up my daughter is in fifth grade but they'll do a cell project perfect and I can't wait to give her the inside scoop on what the mitochondria really looks like but yeah you know it's funny we grew up with these images you know that were fairly outdated but they just sort of still persist in textbooks and animations but we have data that show it's different and then these are the two questions I ask every guest so first is there a memory from high school science that you could share something that impacted you something that stays with you today I think if I had to go back far enough I think it was my first I was always fascinated by anatomy and physiology so it would have to go back to when we did our initial dissections when I was in middle school it frogs and fish and cat and many different things and it always struck me how amazingly organized the body was and then being able to look at it through a microscope to really see what's what we can't see otherwise and just stunned by what I was seeing through the microscope and in the body you know how the blood vessels connected and and the brain communicated the nerve innovation things like that always intriguing to me and it's probably why I've stayed with animal research both because you know I value the every life of an animal of course we consider and we take the utmost care with them but they really have allowed us to their credit to understand how the body works so I would say I'd go back to those hands on days where I really could see and touch and look what I was what I was what I was learning about otherwise in a textbook and then my last question is what advice do you have for high school students today who are interested in studying science? I would probably use the words that you probably hear a lot stay curious ask questions collaborate is a big one never never never think you're in this alone work together you're gonna learn you're gonna get much further in your science career any career by working together as a team there's never a bad question I talk to I say that to my students and my fellows in the residence ask any question there's never a bad question the only bad question is when you don't ask it brings and bring a sense of humor I mean if there's one thing that I always try and and buy if I can is is nothing's ever that bad you can always you can always laugh at mistakes you can find ways to just bring humor into things from a more practical standpoint realize that biology is not just research and and becoming a doctor right that's what people and or a nurse those are wonderful careers but there's much more to science you could be a you could be in drug development we've touched upon that we could be in patent law we didn't touch upon that but there's a whole field that needs scientists and understanding scientists the environment of course sales and marketing we can't get drugs out there if you don't good at understanding the science and how to sell sell something and make sure someone knows how to use it media is a wonderful place for media in science we're doing it today right organizations like the FDA you can work for the FDA and decide on what drugs or do or do not or devices do or do not make it to human therapy and of course the one that I need all ultimately is the National Institutes of Health right those in part of the defense these these are areas where the science is desperately needed continues to desperately be needed and it's not just that you're going to be doing test to work or surgery on a patient there's a real need for science in everything we do and so open your eyes expand your opportunities and see that there's ways to use science in numerous ways that are impactful including doing wonderful podcasts like this or nerds as we like to call ourselves. Oh thank you I think that's such a good point I do think that there's such a when people think of you know science and I want to be a scientist there's still a little bit of like someone hunched over a lab bench like pipetting into a test tube and as to your point that's that's a small fraction of all the career possibilities yeah exactly I think I think that's the thing that you realize that you get fit into these narrow roads of I must do science so I can become a doctor but yeah that's great don't get me wrong but there's so much more to it even if you if you become a doctor doesn't mean you have to see patients there's a need for counters in so many other areas right so it's right it's really a lot of fun wow well Leo this was such an interesting conversation and there's so many I can't wait to follow where this research goes I think it makes me really excited to think about carbon monoxide as part of patient care in many different ways going forward and thank you so much for sharing all this with us today absolutely my pleasure and I hope to hear more and be able to tell you more about it in the future yes be great to have you on again that was Leo Utterbine talking with us about some of the unexpected benefits of carbon monoxide in two weeks we'll have an episode featuring two scientists who have looked at how a substance made up of repulsive particles expands spoiler alert it's not what you think one of the scientists writes ever found yourself surrounded by a repulsive crowd that took forever to expand tune in in two weeks to hear more about this work thank you for tuning in to today's episode of science fair please rate and review the episode on the podcast app of your choice see you next time