Hello, I'm Dr. Matthew Williams from the West Virginia School of Osteopathic Medicine at Lewisburg, West Virginia. Today I'm going to share with you this research project that my wonderful OMS3 student Jackie Botafuco did last summer and then has continued to work on throughout her second and into her third year. I titled this Development of Novel Biotherapeutics for Radiotherapy-Induced Dysbiosis. So when Jackie came to me originally, she had a interest in microbiology and also a future career goal potentially of going into radiology. And me being a microbiologist, I was like, well, how do we tie together radiology and microbiology? And so I sent that back to Jackie to figure that out. I said, what do you know about what happens with the microbiome and radiation therapy? Because again, that was something Jackie was interested in, in potentially becoming a radiologist. So I really personally knew little to nothing about that. So I put it on Jackie to go do some searching, lit reviews, and figure it out. What she found is that, you know, radiation therapy is very common in the rehabilitation plans for many chronic diseases. There are a lot of studies that show that the radiation does have an impact on the microbiome of humans. So a lot of times when we think about microbiome, we're thinking of like gut microbiota. So there's a lot of research out there into that, how radiation therapy can affect the gut microbiota. So there's many cases of patients that when they're going through radiation therapy, they may get some gastrointestinal distress, a lot of diarrhea, a lot of issues with their bowel movements. So there's quite a bit of information out there on that. And some more recent studies showing that there are actually alterations in the bacteria that are in the intestines of these patients undergoing radiation therapy. But keep in mind, it's not just the GI tract that can be impacted. You know, obviously skin surfaces can be impacted as well. And there's a lot less research into that than there is into the gut microbiome in any regards, but especially in regards to radiotherapy. So what we do know with radiotherapy is that it can induce this dysbiosis state. So as I mentioned, there can be multiple issues with like gastrointestinal distress. There's some patients that report some skin lesions or other issues. And many of these can be due to that microbial dysbiosis. So what is dysbiosis? So let's compare, contrast that to eubiosis. So eubiosis is when you have a nice healthy microflora, and it's been found that that helps provide some protection against injury. In this case, again, as I mentioned, many people are looking mostly at the GI associated microbiota. But again, we know that if you have a nice healthy microbiome, that can help protect you against the negative impacts of radiation therapy. Again, that can be potentially extended to things like skin surfaces as well. Again, if you have a perturbed microflora, you may have increased sensitivity to radiation injury. What does the microbiome look like in eubiosis versus dysbiosis? Again, the definition of dysbiosis is that there's less microbial diversity and altered ratios. Again, that microbial balance is getting altered. That's what I mean by dysbiosis. So we know that that's happening with radiation therapy. What's interesting is that the efficacy of radiotherapy has also been shown to be impacted by the microbes that are there. So if somebody is in dysbiosis, if they have the wrong microbes, then it's been shown that radiotherapy may not be as effective. So again, the efficacy of radiotherapy can be impacted by the microbiota. So again, we want lots of nice healthy good microbes to help protect us from the detrimental impacts of radiotherapy, but also the good healthy microbes can help increase the efficacy of radiotherapy. So with this, Jackie wanted to look a bit further into this. She's very much a self-starter and interested in this project. So she wanted to take some of what our lab does and where we look for novel biotherapeutic microorganisms. And so she started screening through many environmental samples. You can see there in table one, where she pulled some of our environmental samples for, again, looking for these novel micro bacteriological biotherapeutics. So she plated out some soil samples, some various environmental samples, and then some water samples. And we did some simple plate-based antagonism assays against our panel of human opportunistic pathogens. You can see those in table two. We have a myriad of opportunistic pathogens we work with in the lab. So what Jackie found was with these initial plate-based antagonism assays, she found several isolates that showed antagonism against her panel of human pathogens. And so after that, we had to determine, are they actually viable to work with? And also, what are they? If we're working with these unknown environmental samples, we want to know what they are. So we sent the unknown environmental samples off for sequencing. So Jackie did the PCR amplification of the 16S gene, and we sent that off to have it sequenced. She analyzed the results to figure out what species she had. You can see that in table three, what species she had with her environmental isolates. So she had some mycobacteriums and cupreavitis, some leucine bacillus, some staphylococcus, some good staphylococci, some commensal ones, not the pathogenic ones, and some erwinia. The other thing we need to figure out is, are they actually viable to work with and potentially make it to the clinic or to the market? So one thing she did is she did some viability assays. So here, she just left them at room temp for two weeks, then four weeks, then six weeks, and all the way up to six months, and we actually still have them sitting there. And she's still checking viability of them. I think right now we're at eight months that they are still viable sitting at room temp. So these are strains that could potentially make it to a market. Again, you don't want something that has to be frozen all the time if you're trying to get it into a marketable product. So again, she just checked them at various time points to see if they're still viable. The other thing she was interested in, again, with the basis of her project being the radiotherapy, is she wanted to expose them to UV as a simple low-level radiation, just to see if they had some resistance to that. And she actually found that though most of them were susceptible to UV, her lysinib bacillus is extremely UV resistant. So she can hit it with lots of UV light for extended amounts of time. And even after six months, eight months of sitting at room temp and then hitting it with UV light multiple times, it is still viable. If you notice here, one interesting thing at table three about lysinib bacillus is that it does form an endospore. So we think some of that endospore formation is why it's UV resistant. But really she's excited about that because lysinib bacillus, as you see over in our discussion, it's already been used as biotherapeutic for many different afflictions. So the fact that she found a UV resistant one that might help in radiotherapy cases is really exciting. Again, we're really excited about all the other isolates we found. And so she's continuing up some other assays trying to figure out, you know, can we kill this with antibiotics? Because, you know, that's obviously something we don't want to be putting these out there if they're antibiotic resistance trained. So she's doing some of those assays and we're really excited about some of the findings she's getting. If you have any questions, feel free to contact me. Jackie's out on rotations, but I'll happily forward any questions towards Jackie and I'm sure she'll get back to you if it's something that I can answer. You can see my contact information there on the bottom right. All right. Thank you.

biotherapeutics

radiotherapy-induced dysbiosis

microbiomes

lysinibacillus

antibiotic susceptibility