Meet the Hosts: The Path to Principal Investigator and Clinical Educator with Dr. Amanda Poholek and Dr. Arvind Srinath

The Poholek Lab | University of Pittsburgh School of Medicine

Pediatric Gastroenterologist Honored with Educator Award | UPMC Physician Resources

Richard King Mellon Institute for Pediatric Research | CHP.edu

Next Generation Sequencing (NGS) - An Introduction | Applied Biological Materials YouTube

What is next generation sequencing? | NIH National Library of Medicine

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Voiceover: This podcast is for informational and educational purposes only. It is not medical care or advice. Clinicians should rely on their own medical judgements when advising their patients. Patients in need of medical care should consult their personal care provider. Welcome to "That's Pediatrics", where we sit down with physicians, scientists, and experts to discuss the latest discoveries and innovations in pediatric healthcare.

Dr. Amanda Poholek: From UPMC Children's Hospital of Pittsburgh, welcome to “That's Pediatrics.” Today we're going to have a very special episode of “That's Pediatrics” called Meet the Hosts, The Path to Principal Investigator and Clinician Educator at UPMC Children's. Hi, Arvind.

Dr. Arvind Srinath: Hey, Amanda.

Dr. Poholek: So this is kind of different and fun.

Dr. Srinath: So who are we interviewing today?

Dr. Poholek: I think we're interviewing each other.

Dr. Srinath: Interesting.

Dr. Poholek: Yes.

Dr. Srinath: Why are we doing this?

Dr. Poholek: So that the listener gets a sense of who we are, how we got here, and why we're the podcast hosts.

Dr. Srinath: Got it. This is a little bit of a different role than we usually have, right?

Dr. Poholek: It is.

Dr. Srinath: I'm not sure if I'm ready for this.

Dr. Poholek: It is. I am also nervous, but I think it's okay. We're going to be fine. We've done this before. We'll be good.

Dr. Srinath: Why don't I just ask the questions and you answer them.

Dr. Poholek: Sure. Let's get started. Let's start that way.

Dr. Srinath: Okay, fine.

Dr. Poholek: That'll work.

Dr. Srinath: Fine. Fine. I'll start. I'll start. So Dr. Poholek, what is your role? Who are you and what do you do?

Dr. Poholek: Yeah, great question. Sometimes I actually find that a little bit hard to answer, but I think in general I would be described as a Principal Investigator. So I think you can also give me multiple other titles, including Assistant Professor of Pediatrics, which is often how I introduce myself as a co-host of the podcast. I'm also a Scientist, I'm an Immunologist. I'm also the Director of the Health Sciences Sequencing Core, which is a core facility that does next-generation sequencing for the entire University of Pittsburgh and even beyond sometimes. So I guess you could say I wear different roles, mentor, teacher, yeah, lots of different jobs. But ultimately I run an academic research lab that is funded by the NIH where we try to understand questions about the immune system.

Dr. Srinath: So you have multiple roles. What do you tell your kids that your role is?

Dr. Poholek: Yeah, great question. I think for the kids we generally say mommy's a scientist or an immunologist.

Dr. Srinath: Nice.

Dr. Poholek: Try to keep it to sort of those. My kids are six and 10, so kind of at that level. But they also understand that we have students, right? So my husband is also in the same role, and we have students, we have postdocs, we host parties at our house and these people come and so we introduce them as the people in our lab. They know we have a lab. They've come to our lab to visit, more so before Covid. But yeah, so they definitely kind of have a sense of us being scientists, but also sort of directing the program.

Dr. Srinath: See, I'm asking all the questions, Amanda, see how this got manipulated in the right way?

Dr. Poholek: I see. I see our [inaudible 00:03:23].

Dr. Srinath: So when you say lab, are you wearing a white coat and wearing goggles and just being in a lab all day, or what do you do?

Dr. Poholek: Yeah, so not anymore. But that certainly was part of my path to get to this job, right? So I guess maybe we can answer that question. How does one become a principal investigator? How did I get to this point? So I guess for me, it started in my undergrad. I did my undergraduate at Fordham University in New York, and I knew I liked science and I knew I wanted to do science in some capacity. I think I initially thought I was going to teach science, and so I started off being a major in biology, and then I kind of realized that one, all of my classmates were pre-med. And no offense, but at the time, being a physician didn't appeal to me.

Dr. Srinath: None taken.

Dr. Poholek: And I kind of started to realize that research was a thing that you could do. And I was in the Honors program and we were required to do a master's, and most of the other science students had actually done some research.

And so it kind of dawned on me that I was going to need to find a lab and do research in order to do my thesis.

Dr. Srinath: Got it.

Dr. Poholek: So I did that and I really loved it. I think it was kind of one of those things where you start doing it and you're like, "Oh wow, this is so much fun." I've always enjoyed doing things with my hands like cooking or fixing things. And so being at the bench and actually playing with science and watching it kind of happen before my eyes was just really magical for me.

Dr. Srinath: That's so cool.

Dr. Poholek: I did a summer in industry, which I also really enjoyed and learned a lot. And then by the time I kind of became a junior, I realized that I wanted to get a PhD.

Dr. Srinath: Got it.

Dr. Poholek: I didn't want to go to medical school like most of my classmates. And I really wanted to sort of understand the basics behind disease. I also was always interested though in understanding sort of disease-based biomedical science.

Dr. Srinath: Got it.

Dr. Poholek: So I started applying for graduate school. I was really fortunate to get into Yale University into sort of an umbrella program, which gave me the opportunity to sort of explore different disciplines, cell biology, cancer, but ultimately fell in love with immunology, and did my PhD at Yale in immunology and sort of with a lean towards cell biology. And then at that point, I think I realized that I wanted to do academia. I wanted to run my own lab. I wanted to figure out a question that really I was passionate about and that I could answer myself using research. And I did a postdoc, which is sort of the next step in that track. I went to the National Institute of Health, did a five, six year postdoc there also in immunology and sort of further refining the type of question that I was interested in learning the technologies that I needed to answer that question, which for me ended up being a lot of next-generation sequencing.

And then along the way, I met my husband in grad school. He and I had to sort of then move together. And then we were really fortunate to be able to move together here to Pittsburgh where we both were able to get tenure track faculty positions to run our own labs and start that process. So in the beginning, I still was at the bench sort of hiring my first technician, setting things up. Now that I have a staff that does all of that part, the PhD students that I'm training, the postdocs in my lab, I mostly just sit at my computer and write grants and write papers and mentor and teach. And I love that part too. I really do. Watching your students grow as scientists is an amazing and wonderful experience that I wouldn't trade for anything.

Dr. Srinath: It just fascinates me because you're constantly thinking about what's the next question? How do I investigate this? Why? Why? Why?

Dr. Poholek: Yeah. No, very much. Very much.

Dr. Srinath: Yeah.

Dr. Poholek: And I like your idea about next question because I think next question, Arvind, what's your job?

Dr. Srinath: That escalated quickly.

Dr. Poholek: You walked right into that one. Tell me about your responsibility here at UPMC Children's?

Dr. Srinath: Yeah, so I guess according to this title of this podcast, I'm what they call the Clinician Educator, which means Clinician. I see patients and I work with patients and families to help with their disease processes. As an Educator, I teach medicine. So more specifically, my role is I'm a Associate Professor of Pediatrics in the division of Pediatric Gastroenterology. I'm going to abbreviate that as GI. And I direct the training program where they call that the fellowship program and help the rest of the fellowship programs as the Associate Vice Chair for education for fellowship training. So I guess that's what I am.

Dr. Poholek: Yeah. Okay. So you're a doctor, you see patients, peds GI, right?

Dr. Srinath: Yeah.

Dr. Poholek: So specializing there. And then when you're not seeing patients, what kinds of things are you doing? Are you sort of trying to figure out what the fellows are up to? How to change the fellowship program? What is sort of like the day-to-day versus what might be the bigger picture goals for that part of your job?

Dr. Srinath: Half of my time is spent working with patients and families, and the other half of my time is development of curriculum of didactics or teaching sessions of helping enhance how well we are educating our trainees, specifically pediatric fellows. And to be honest, trying to find ways that I could, taking a page out of the yearbook, put what I do onto paper to help disseminate things that we're finding to help enhance educational techniques across the nation if possible.

Dr. Poholek: So can you give me an example of something that you were part of or that you instituted as a clinician educator for the fellows here that you're really proud of or that you feel like was something that was important to you?

Dr. Srinath: No. No. Okay.

Dr. Poholek: I don't believe you.

Dr. Srinath: So actually I'm really proud of something that came from within. So we have a fellows council, which is a group of fellows that meets together to identify areas for enhancement across our fellowship programs. And I work with them. And a couple years ago, we all designed a needs assessment. So we sent a survey out to figure out how we can enhance the fellowship curriculum. And out of that needs assessment came out this idea to hold yearly fellow retreats for each level of fellowship. Now I'm getting a little bit technical here, but I think it's really cool. And part of training to be a pediatric gastroenterologist or cardiologist or anything about pediatrics is after college you go to medical school, then you train for three years in pediatric residency, and then you have to do an extra three years of training in whatever you want to do within pediatrics, GI, allergy, et cetera.

That first year is predominantly working with families. And the second two years is mainly devoted to really doing what you do is to really figure out how we can ask a question and figure out how we can really think as a scientist. So we designed yearly retreats. So the first year we put together a few sessions to really how they can enhance who they are as clinicians and having difficult conversations to really get at how they work, how to work well as a first year fellow? And then the second two years were geared towards how to get a job? So how to brand yourself to make sure you identify areas where you can enhance positions that you're applying to? And third year is how to take the next step, what's it as being a junior faculty and how to really move further in your career once you become a junior faculty? Honestly I felt it was really well received and it felt like we took what was needed from within and came up with something.

Dr. Poholek: Oh wow. That's so cool. Yeah. I really am interested in this idea of how you brand yourself as a physician, because this is something that we have to think about as scientists as well. Very much so. How are you going to bring something to the institution that's distinct? So for example, when my husband and I were searching for jobs, we both had a research interest. We're both immunologists. I'm very interested in sort of the cell biology of the immune system, how cells make decisions. I use a lot of next-generation sequencing.

My husband Tim Hand, he does a lot of microbiome research, mucosal immunology. And so we had to find a place that kind of wanted both of those interests, right? You want to find a place where you fit in to the group that you're bringing something to the table that everyone's excited about and that you can also grow. So I have no concept about how you do that as a physician, right? Because my scientist kind of idea of being a doctor is like, "Yeah, you see patients and there's diseases and that's it." It's more of my experience as a patient or as going to the doctor. So can you comment on that? What does that look like for fellows becoming faculty?

Dr. Srinath: It's like how do you find who you are as a person, really?

Dr. Poholek: Yeah.

Dr. Srinath: And because the majority of our lives are really spent doing work, right, so how do we make that to the point where we're finding purpose that helps with wellness, that helps with a lot of things? So I think it's helpful. And I promise to be two minutes. I'm a little bit chatty, Amanda. You're more direct to these questions, so you can just cut me off. But I grew up in a small town. I'm 5'8", right, so I am not the tallest person, but I tried to play tennis and after I hit puberty I realized I wasn't going to be the tallest person. So genetics probably weren't helping me. But I like teaching tennis and I ended up teaching tennis for a couple years. Well, I probably should have been doing what you were doing in college, doing a lot more scholarly work.

But I loved it for seeing people contact a ball for the first time, or master a stroke, or really master a strategy. I swear there's a point to this. That helped me realize that, "Yeah, I like development, I like pediatrics." I also liked that kind of helping people find their spark and that self-efficacy, having that spark and perpetuating that moving forward, moving forward. And fast-forward, I came into fellowship and after that first year of constantly teaching, being on the floors, we are asked to find a research project and to move forward with that. And I did and it landed a small grant when I was a faculty member.

But then two years into faculty, I realized, what do I like about this? I like the dissemination, I like watching people as they're like, "Oh my gosh, this is so cool, right, and I'm going to try this." And I realized there's more of the teaching than the actual what you do is the questions and the finding. And then I went to my boss at the time and said, "I'd really like to direct the fellowship program and just do that. I want to get a master's in medical education to help put my best put forward. And I realized that through that master's, what I really loved is designing curricula, designing how can people really become an expert in what they're doing within their fellowship programs or even beyond that too? And that's how I felt like I kind of branded myself in addition to helping find that spark.

Dr. Poholek: Yeah. So through tennis, you found your passion kind of for teaching and mentoring and then utilized that to sort of position yourself and brand yourself as a clinician educator. Amazing.

Dr. Srinath: I swear I didn't give her those lines. She figured that out herself, folks. So, yeah, exactly.

Dr. Poholek: So I want to actually quickly jump back to what you said about getting a master's. So was that something that the hospital system helped you with and supported you? Is that something that's available here?

Dr. Srinath: I think the beauty between both of us. I think both of us have utilized this amazing system to get to where we are. To be honest, I'm intimidated [inaudible 00:15:35] so what you do? And I still have to figure out what next-generation sequencing is. So I have that as particular to ask you in a second. The University of Pittsburgh School of Medicine, which is affiliated with us, offers degree programs, namely certificate programs and degree programs where you get full on masters. And one of them is in medical education. And the beauty of it is is that they recognize many of us are actually seeing patients during the day, for example and forming other clinical duties. And these classes are superbly structured to be at times which are not necessarily the typical work hours when we're in clinic. So it was actually pretty easy to do.

Dr. Poholek: So it's not like you have to be a full-time doctor and get the degree. They actually are able to carve out some time for you to get the degree in addition to obviously still doing a lot of being a doctor.

Dr. Srinath: Actually I was still a full-time doctor and got the degree.

Dr. Poholek: Oh, wow. And they're just making it so that it's possible to do both.

Dr. Srinath: Exactly. And when you're doing something you love...

Dr. Poholek: And that's funded, right?

Dr. Srinath: So I had to pay for it.

Dr. Poholek: Oh.

Dr. Srinath: We had an employee discount, so it was definitely manageable to do that.

Dr. Poholek: And so the masters is through Pitt or through?

Dr. Srinath: It's through Pitt School of Medicine.

Dr. Poholek: Oh, neat.

Dr. Srinath: Yeah.

Dr. Poholek: Oh, that's really cool. And do you feel like that really helped guide you in sort of what you wanted to do next? How instrumental was that for you in the next step?

Dr. Srinath: Hugely.

Dr. Poholek: Okay.

Dr. Srinath: Because I think when you realize what you want to do with your career and you're taking these courses later on, you're doing it from within. I didn't have to do the masters, I didn't have to do any of it. But I'm doing it because I want to become a better teacher, better at what I want to do long term. So when you're primed to thinking, "Hey, I'm interested in curriculum development," I was focused on trying to find a research, a mentor there who's involved in innovative ways to foster, to put together curricula, which was my main goal. And even in the courses, you realize whatever you're taking in, you're going to extrapolate the next day with trainees.

Dr. Poholek: Right.

Dr. Srinath: So now it's my turn because I feel like I've been talking too much and we need to turn the table because you're asking amazing questions. And I'm interested, I haven't even figured out what next-generation sequencing is yet. So tell me what about this hospital system brought you and your husband, a power couple, to this place? I mean, full disclosure folks, Dr. Poholek's CV is about 40 pages long and she's done 20,000 things. She's super underestimating, underselling, what she does. So what drew you here?

Dr. Poholek: Yeah, that's a great question. So I would say that there's a number of things that drew us here, but probably at the top of the list was the R.K. Mellon Foundation for Pediatric research. This was a foundation that was established specifically with the goals of aiding and supporting research in pediatrics. At the time that we were recruited here, the person that was running that program had identified microbiome as something that they really wanted and an immunologist on their team. They specifically recruited researchers who were junior. There's an idea, and I think it's some validity to that idea that younger scientists as they're emerging, have a lot of creative ideas, very innovation forward. And so the main goal was to recruit junior investigators that were going to be very innovative and really move pediatric research into a direction that you couldn't do otherwise without hard money support, right?

And so that was the setup of the Foundation. And they recruited my husband to be part of that. Over the next couple of years, they actually ended up transitioning the program to be broader. So now all of us who are tenure track investigators in pediatrics are part of that program. So technically I'm a scholar in that program. We're all scholars. And they've really extended the network so that you can apply for grants that are only available to the scholars. So you're basically competing against your friends and your colleagues, but also there's only a few of us. So there's limited competition for resources, which is always helpful. They have pilot awards, they have innovation awards. Actually, one of the things that's really helpful is they have money specifically for the trainees in our lab to support their salaries.

So that just means you get another person in your lab that you don't have to support off your grants, which is really helpful. One of the things that I think has been incredibly helpful as a junior investigator is a grant writing program. So when you want to submit your first big grant, which is called an R01. These are very hard to get. It depends on the specific institute you're applying to, but let's say somewhere in the ballpark of trying to get into Harvard every time over and over again, and you need more than one. But when you're trying to do your first one, you get a bit of a break. And so they have set up basically an internal program where you write your grant and you send it to the people running the scholars program.

You can help them identify potential reviewers with the right expertise. They actually will pay them to do that job to review your grant. Not a ton of money, but just enough that when they say, "Yes, they'll do it," they'll actually do it. And then within two to three weeks, a very short turnaround time, they'll read your whole grant. These are all people who have grants and who serve as reviewers on grant study sections. So they kind of know what it should look like and they provide feedback. And it's invaluable because it's almost like you get to get up to bat and take a swing before really going out there. And so the first one I submitted internally was incredibly helpful. I think I would've sent a grant that probably would've done very poorly, but instead I just sent it to the internal group who said, "Hey, this is really starting to look good, but you need these things. You need more feasibility data. You need to restructure this part of it. This part's not clear."

And then that gave me an opportunity to spend three or four months just sort of generating that extra data that was really going to make it go to the next level. And that gave us the opportunity on the first submission to at least get a score. Didn't get funded, but at least we got a score.

Dr. Srinath: That's huge.

Dr. Poholek: And then we were able to revise that and resubmit and get funded on the second go. And I think it's not uncommon for junior investigators to have to submit 2, 3, 4 times, sometimes even more, to get their first grant. Having that first round be sort of internal, but still really high level. 'Cause these are the people who do the grant review. It just really made my life a lot easier when I had to actually do it for the first time. So that part of the program, I think is really invaluable. And you can continue to use it throughout. For my second grant, we used it again. It was great. It was really helpful.

Dr. Srinath: That's a huge infrastructure we have. Can I ask you a few questions about that if that's okay with you?

Dr. Poholek: Yeah, absolutely.

Dr. Srinath: So it's almost like a, I don't want to say academy or institute, but literally the infrastructure here is basically, guess what? We got you, we're working together as a team. We're going to prepare you and have the scrimmages and have the dress rehearsals, and guess what? And then you're going to have the opportunity to go out there so it doesn't feel like deer in the headlights. How many programs are like this across the nation?

Dr. Poholek: There's not a lot of examples. I mean, there's a few. Sometimes you'll find institutes or you'll find smaller programs, but there's not really anything like this that I'm aware of. It doesn't mean it doesn't exist, I just don't know about it. But there aren't that many that I know of that are a group within a group. So my primary affiliation is at the University of Pittsburgh, and I still have all of the sort of benefits that the School of Medicine provides to everybody who's a tenure track investigator, of which there are many, but sort of within this R.K. Mellon Foundation, the Scholars program, which is run by the way, by Dr. George Gittes. It really, I think, creates this smaller culture of people that are nearby you. And that has a more focused sense of, "Okay, what can we do for our junior investigators that are here? What resources can we provide? Physical resources."

Another thing they recently came out with is when you first get started, you don't have grants. You do have hard money from the institution, but you want to do some big experiments to get started. And those often are expensive from cores, for example, next-generation sequencing. And so they just created a new program where if you want to do a core facility at Children's that will help you generate data for a grant they king of give you a subsidy rate on that, they give you a break, or they throw extra money at you just specifically for that purpose. So these kinds of small things can actually make a big difference, because if that's a critical data set that you need to generate, but you're lacking an extra 10K to do it. Okay, if the foundation can throw a little money at you so that you can get that started.

And that's really what the foundation is set up to do. They're set up to sort of kickstart you. You could think about it as a kickstarter fund. Yeah?

Dr. Srinath: Got it, got it. Okay.

Dr. Poholek: So they kickstart you and give you that initial financial resources and space and environment to get that first set of data out there and say, "Okay, I came with an idea. Here's the bit that actually proves my idea might be right. And now I can use that to apply for a real grant from the NIH, and that's now what's going to start to keep me going." Success begets success. It becomes a cycle of, you take the NIHs money, you make data, you turn that into papers. That's the product of science is publications, right? It's knowledge. That's what we're producing is knowledge. And so that knowledge goes into publications, that's our product. And then we use those products to get more grants and say like, "Okay, look at all this cool stuff we discovered. Now I have more ideas. Can you give me money for those too?" That's the cycle basically of being a scientist.

Dr. Srinath: That's so fortunate. And that's great because it sounds like at least from the outsider perspective, that the activation energy to get things going with an idea is huge. And this infrastructure you're talking about really helps you get over that activation energy to get into the NIH funding and other competitive funding, which to be honest, is probably not as easily accessible outside of this institution. My question is, where is this money coming from for the R.K.? Can you tell me about the Foundation?

Dr. Poholek: Oh yeah. I actually probably should know more than I do. I mean, my understanding is that it came from the R.K. Mellon family who wanted to do this. And they've set aside a certain amount of money that I believe is probably in some sort of endowment or situation. And that that is then managed by the institution by UPMC Children's to make sure that that money sort of continues. And I think there was an initial sort of promise, and then I think they go back and there's a committee that comes in and oversees it and all of the appropriate oversight that one needs to make sure that it's being properly managed and properly distributed. And this gets outside of my area of expertise as a scientist.

Dr. Srinath: Oh, totally. Yeah. Yeah.

Dr. Poholek: But yeah, it's really just the Mellon family and their generosity. And I mean, I think those of us listening who live in the Pittsburgh area know that there's a number of these prominent families that have been incredibly generous to the hospital system that make it possible to do all kinds of things that you couldn't do elsewhere.

Dr. Srinath: Yeah.

Dr. Poholek: Absolutely critical. And philanthropy is a small but significant part of, I think the UPMC Children's Hospital system for sure.

Dr. Srinath: That's amazing.

Dr. Poholek: Yeah.

Dr. Srinath: That's amazing.

Dr. Poholek: Yeah. I want to talk a little bit about kind of how things have changed from the time that you were a fellow. I guess maybe, I know in my mentoring, a lot of what I've learned about being a mentor came from my experience with the mentors that I had. And you see things that you like, you see things that you want to do different, that's normal. I'm curious, I think both about your personal experience being a fellow, because I think you did your fellowship here.

Dr. Srinath: Exactly.

Dr. Poholek: And then becoming a fellowship director and what that was like? And also just more broadly, how has being a physician, being a pediatrician, changed from the time that you were a fellow to sort of the fellows that you're training now?

Dr. Srinath: I think it really gets on your theme of branding, branding, finding your passion, finding your purpose, which promotes scaffolds into promoting resilience and helps people with career longevity and career satisfaction too. Details wise, I mean, I've been here and there between institutions, so I'm not sure if I said this and I keep on talking, so I might have just said it. I'm going to repeat it. So anyway, is I grew up in a small town in Connecticut, went here between Johns Hopkins for college, here for medical school, back there to Hopkins for residency, and came back here for fellowship and stayed on [inaudible 00:28:28]. We all know Pittsburgh infrastructure pulls you in and the city is just second to naught. The reason why I bring that up is because I think there's some themes that were there when I was training which are a little bit different now, which what I really try to focus on for program development here, and that is deliberate practice, finding your passion in what you do.

I think it was a little bit unwritten, but kind of understood that when we were in residency or even fellowship is the latter part of your fellowship is devoted to scholarly activity. Okay? And what that scholarly activity did you for your career was a little bit up in the air in terms of what fellows I think understood. So what I sometimes saw is some fellows doing bench research during their second and third years of fellowship, but then when they went to practice, that wasn't any part of their practice. And the purpose of the bench research were a little bit unclear.

Dr. Poholek: Yeah. I feel like I've come across some of those fellows in my time training.

Dr. Srinath: Right. And I think what we've since really tried to promote is guess what? The purpose of your training is to figure out what you want to do. What do you like within pediatrics, within pediatric GI, within pediatric endocrinology that you can use to extrapolate? The scholarly time during fellowship should be devoted to what you want to do, whether that be bench research, clinical research, or medical education, quality improvement, right? Those are other areas that I think weren't as tackled in the past, but there was interest.

And within medical education, there's curriculum and development, there's a lot of areas there. I mean quality improvement, there's applying quality improvement strategies and topics to every single field out there. And same thing within clinical bench research, if I did clinical research during my second and third year of fellowship, utilizing that to brand who I am. I'm a person who did clinical research in terms looking at functional pain in patients with Crohn's disease, right? Gee, that helps me because maybe I have a clinical niche in inflammatory bowel disease, particularly the psychosocial aspects of it. And that helps me brand myself as a person, helping find [inaudible 00:30:48] and helped me funnel who I'm seeing trying to help most as a clinician.

Dr. Poholek: Oh, okay. So that makes a lot of sense. And so this is interesting to me because one of the things I get to do is interview prospective fellows for the pediatric rheumatology program, which is the division that I'm in. And it's interesting because you of course come across some fellows that know exactly what the plan is, they know exactly what they want to do for their research time. Sometimes it's bench research, sometimes it's not. But then you also come across the ones that are less sure. They're interested in exploring options. They don't really seem to know yet what they want to do.

And that first year of fellowship is heavily intense, right? Oftentimes you're moving to a new hospital system, so there's just the ins and outs of a new job that you have to get used to. And also you're seeing a ton of patients and learning a lot of your clinical education in this new subspecialty that you've signed up for, right? And then somehow also in that time, they're supposed to figure out their path, their passion. So how do you help the people that don't know what they want to do? They know they love being a doctor, they know they love rheumatology or GI, but that scholarly project is maybe they know bench research is not for them because most people seem to have a sense of that right up front if it is or isn't. I'll say that. So how do you direct them?

Dr. Srinath: My wife's an attorney and she always tells me when I'm talking to people, "Ask questions, ask questions, gather information," and I ask a lot of questions. There's multiple ways of doing this. But particularly I personally try to focus on if it was a really difficult day, a stressful day. What drove you? What was the highlight?

Dr. Poholek: I see. I see.

Dr. Srinath: Right?

Dr. Poholek: Yeah.

Dr. Srinath: Gee, there was a pathophysiology, but this condition was really, really interesting to me.

Dr. Poholek: Okay. That's insightful, right?

Dr. Srinath: Yeah. Helping this patient and family through this difficult conversation was super helpful. Or I guess teaching these residents how to do acts really was a highlight of my day, or recognizing there was a process that could be improved that really got me going. I wish I had the time to do it. Well, there you go. Right? That's one of many strategies.

Dr. Poholek: Neat. So, Arvind, what's next for you? Where do you see the next steps of the fellowship program and where do you see the infrastructure of UPMC helping you with that vision?

Dr. Srinath: Yeah, so I think what's the beauty of the UPMC infrastructure is that not only do we have great clinical teaching and great clinical and research mentors, I'm looking at one right now, but I think we have a great infrastructure to help people kind of disseminate what they want to do. And I think that's the next step. We've put together a pediatric fellow scholarly toolkit on the website to really help give fellows even more tools to help pursue the academic paths for whatever area they're interested in in hopes that granted there's an interest, but to take a page out of the yearbook to see if they can get funding to help really dig deep into those interests and help others along with that, because that's what drives us.

And between what we have in Children's, the training grants we have within our institution and outside our institutions and the grant funding that we get, and also utilizing the amazing providers and scholars and mentors that we have, I think that's the next step. And really finding how well we are doing along those lines and how we can improve beyond those lines too.

Dr. Poholek: Yeah.

Dr. Srinath: So how about you? What's the next step for you? First of all, what's next-gen sequencing?

Dr. Poholek: What's next-gen sequencing? Yeah. No, that's a great question. So it's actually kind of a boring answer. Broadly speaking, it's a system. It's a set of machines. It's a system that allows you to understand the bits of DNA massively in parallel. So instead of taking one small piece and sequencing at one nucleotide at a time, which is sort of how Sanger sequencing works, you basically can layer a bunch of sequencers together onto a glass slide, and it sort of amplifies. The machine will rapidly amplify those in place on the slide and then also give you an opportunity to sequence them in parallel at the same time. So you can get all of this information about sequences, whether that's derived from a person's genome, from DNA sequencing, whole genome, whole exome, whether that's RNA that you took from a cell and converted it into cDNA and then went on and did your sequencing so we can understand all the genes that are being expressed in a cell, which is called transcriptome analysis.

I mean, it's amazing what's happening in NGS technology. Being a core director is just a small part of my job here. But actually it's really fun because you get to see all these different really cool projects that people bring from all over campus. And sometimes we're doing projects on pediatric urine and sometimes we're doing research animals that are in neurology. I mean, just the breadth is tremendous. It's tremendous. And it never ends in terms of the coolness of the things people are doing. In terms of what's next with that, I actually am very passionate lately, and I'm using this both in my own research and I'm very excited about bringing this to UPMC and to the University of Pittsburgh is what's called spatial transcriptomics. So for a long time, if we wanted to understand what genes were being expressed in a cell, which is sort of the mRNA, you had to extract that information from the cell.

So initially we would do this in bulk. So you would take a group of cells in a tube that you would extract from a tissue or from a sample, and then you would just hope that you could figure out what the cells were. Eventually, this technology transitioned to single cell, right? So now there's these machines that can take individual cells, put them into these little cool oil droplets together with all the things you need to get the information out and make a library. And then those libraries go onto the sequencer. And so you can actually get the transcriptum of each individual cell, which has totally revolutionized our understanding of biology. It's hard to really understate how much that changed the game. But even still, you had to take the cells out of the context and you had to put them in single cell suspension and get them through the machine, which as an immunologist was dead easy and we really leveraged this technology.

But for many people, that was harder, right? And sometimes not possible. So spatial transcriptomics basically takes sort of what the pathologist knows and loves, which is sections of a tissue on a glass slide, and now derives the RNA from that in the context of its spatial tissue. So it's almost like, imagine you took your tissue on a slide and you gently permeabilized it and let the RNA fall out and it sticks to the glass slide, and then you basically do the sequencing on the slide. Not exactly, I'm sort of generalizing here, but you basically do the sequencing mapped back to the slide. And then you can take the data and say, "Where was the cell in the tissue?" And it started off at a resolution that wasn't quite single cell, and it's really moving quick.

It's moving very fast to the technology. And so even in a short couple of years, we're now almost down to a technology that's single cell level. Now, the size of the tissue that you can put on that space isn't quite that big yet. But these things, again, are rapidly improving. And even beyond that, there's some new technology that's coming out that doesn't use sequencing actually. It's a C2 hybridization system. So basically, we know most of the genes, right? We have the sequence of the genome. You can basically make probes that match those sequences and fluorescently tag them. And again, through a series of cool technology, you can layer those together in groups and then take pictures of them in the tissue. So this is like an RNA and C2 hybridization, but massively paralleled in a spatial context. And so those machines have just come out and I'm desperate to get one.

And so I'm in the process of putting together a grant to try and get funding for that piece of equipment for our core, because I think already we're seeing just the technology that we're already doing, which is sort of the one on the glass side. We've been doing that now for over a year. And I mean, everybody is just coming to us and asking, "Can I do this? How quickly can you do this for my project?" But I really want to build out sort of the spatial transcriptomics for our core, because I think this will really be the next big step for not just my research, but everybody's research here. And so I'm super excited about seeing what people will do with that and just watching that grow. So that's definitely an area of growth that I'm excited about that UPMC has been very helpful in continuing to support, and they've definitely helped us get some of the pieces along the way that we needed in order to implement this program. Yeah.

Dr. Srinath: I have about 30 things I need to go look up based on what you just said right now. And I feel like that escalated super quickly to the [inaudible 00:40:02].

Dr. Poholek: I know. I feel I was like, I'm going to get real technical real fast. I tried to keep it very broad.

Dr. Srinath: It was fascinating. No, in a positive way. It's totally super fascinating. I was like, "Oh my gosh, what is she doing?"

Dr. Poholek: I mean, from a clinical perspective, it's so exciting because there are so many clinical samples that we've already got on slides that we know are important from patients that have all kinds of really fascinating things going on. You could even think about it potentially diagnostically, right, if you are for it. I mean, there's just so many things that I think we're going to be able to do, and the technology's moving so fast. It's just really cool. I'm really excited for the next 10 years because I know how much we've done in 10 years from the time that I was but a student, and just imagining what it's going to be is so exciting. It's so exciting.

Dr. Srinath: So we're going to follow up to this podcast in 10 years when Amanda come and says...

Dr. Poholek: Gosh, I'll give you manual updates if you let me.

Dr. Srinath: Right, right, right. Then after the list of things to look up. My gosh.

Dr. Poholek: Well, that was a lot of fun. So hopefully this has given all of our listeners an idea of the path to becoming a principal investigator or a clinician educator at UPMC Children's Hospital.

Dr. Srinath: And I'll second that, was how to brand yourself. I think both of us have kind of shown you what path one can take to brand yourselves and some techniques you can all think about depending on what you want to do.

Dr. Poholek: Yeah, absolutely. Such a good point. Fantastic. All right, well, I'll see you next time.

Dr. Srinath: Thank you again.

Voiceover:  You can find other episodes of That's Pediatrics on Apple Podcasts, Google Podcasts, Spotify, and YouTube. For more information about this podcast or our guests, please visit chp.edu/ThatsPediatrics. If you've enjoyed this episode, please be sure to rate, review and subscribe to keep up with our new content. You can also email us at podcast.upmc@gmail.com with any feedback or ideas for topics you'd like our experts to cover on future episodes. Thank you again for listening to, That's Pediatrics. Tune in next time.