#024 - Navigating the Neonatal Immune Landscape: Insights from Dr. Bunmi Olaloye

Hello friends 👋

In this episode of At the Bench, hosts Misty Good and Betsy Crouch welcome Dr. Bunmi Olaloye, a neonatologist and physician scientist at Yale School of Medicine. Dr. Olaloye shares her path from an early interest in engineering to focusing on necrotizing enterocolitis and spontaneous intestinal perforation in preterm infants. She discusses her research into early life immune development using mass cytometry and single-cell RNA sequencing, revealing distinct immune signatures that could shape future treatments. 

The conversation also explores the challenges of building an independent research career, the importance of mentorship and team science, and the limitations of current animal models. Dr. Olaloye highlights the potential of organoid systems for studying human-specific questions and the protective role of breast milk in modulating immune responses in NEC. Her story offers both scientific insight and genuine inspiration for anyone navigating the path from clinical training to independent research.

Link to episode on youtube: https://youtu.be/Ia9NXiEDtnI

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Short Bio: Dr. Bunmi Olaloye is an Assistant Professor of Pediatrics in the Division of Neonatology at Yale School of Medicine. She received her M.D. from Rutgers New Jersey Medical School, completed her residency in Pediatrics at the University of Texas Medical Branch in Galveston, and her neonatology fellowship at the University of Pittsburgh Medical Center, Children’s Hospital of Pittsburgh. Her research interests include deciphering the role of immune dysfunction in neonatal intestinal diseases--necrotizing enterocolitis (NEC) and spontaneous intestinal perforation (SIP)--to identify biomarkers and therapeutic targets and improve outcomes for patients.

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Featured Manuscripts From Dr. Bunmi Olaloye

A single-cell atlas of circulating immune cells over the first 2 months of age in extremely premature infants.

Olaloye O, Gu W, Gehlhaar A, Sabuwala B, Eke CK, Li Y, Kehoe T, Farmer R, Gabernet G, Lucas CL, Tsang JS, Lakhani SA, Taylor SN, Tseng G, Kleinstein SH, Konnikova L.Sci Transl Med. 2025 Mar 5;17(788):eadr0942. doi: 10.1126/scitranslmed.adr0942. Epub 2025 Mar 5.PMID: 40043141

Single-cell atlas of the human neonatal small intestine affected by necrotizing enterocolitis.

Egozi A, Olaloye O, Werner L, Silva T, McCourt B, Pierce RW, An X, Wang F, Chen K, Pober JS, Shouval D, Itzkovitz S, Konnikova L.PLoS Biol. 2023 May 19;21(5):e3002124. doi: 10.1371/journal.pbio.3002124. eCollection 2023 May.PMID: 37205711 Free PMC article.

Single cell analysis via mass cytometry of spontaneous intestinal perforation reveals alterations in small intestinal innate and adaptive mucosal immunity.

Olaloye O, Eke C, Jolteus A, Konnikova L.Front Immunol. 2023 Feb 7;14:995558. doi: 10.3389/fimmu.2023.995558. eCollection 2023.PMID: 36825028 Free PMC article.

CD16+CD163+ monocytes traffic to sites of inflammation during necrotizing enterocolitis in premature infants.

Olaloye OO, Liu P, Toothaker JM, McCourt BT, McCourt CC, Xiao J, Prochaska E, Shaffer S, Werner L; UPMC NICU Faculty; UPMC Pediatric Surgery Faculty; Gringauz J, Good M, Goldsmith JD, An X, Wang F, Snapper SB, Shouval D, Chen K, Tseng G, Konnikova L.J Exp Med. 2021 Sep 6;218(9):e20200344. doi: 10.1084/jem.20200344. Epub 2021 Jul 16.PMID: 34269788 Free PMC article.

Role of Nutrition in Prevention of Neonatal Spontaneous Intestinal Perforation and Its Complications: A Systematic Review.

Olaloye O, Swatski M, Konnikova L.Nutrients. 2020 May 8;12(5):1347. doi: 10.3390/nu12051347.PMID: 32397283 Free PMC article.

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The transcript of today's episode can be found below 👇

Misty Good (00:01.795)

Hello and welcome back to At the Bench, the Neonatal Physician Scientist Podcast of the Incubator, where we are incubating discoveries. I'm Misty Good, a neonatologist scientist and division chief of neonatology at UNC Chapel Hill. And I have the absolute pleasure of co-hosting with Dr. Betsy Crouch. Betsy, do you want to introduce yourself?

Betsy Crouch (00:22.338)

Hi everyone, happy new year. I'm Betsy Crouch and I'm an assistant professor of pediatrics and stem cell biology at the University of California, San Francisco. Also very happy to be here and happy to get to have a nice discussion with our guest today, Dr. Bunmi Olaloye. She is an assistant professor of pediatrics in the division of neonatology at Yale School of Medicine.

Bunmi, thank you so much for joining us. Do you want to provide a brief intro to you and some of your passions to get us started today?

Bunmi Olaloye (00:58.795)

Yeah, hi, Misty and Betsy. Thank you so much for having me on the podcast today. I'm super excited to talk to you about my journey to becoming a neonatologist and physician scientist. As you mentioned, I'm currently a junior faculty at Yale and I'm transitioning to independence while working with my mentor, Dr. Liza Konnikova, who is an immunologist and neonatologist here in New Haven. And like a lot of physicians, I was interested in a career in medicine pretty early on, inspired by my dad, who was an orthopedic surgeon. But then in high school, I took a little detour and was sort of interested in engineering after an amazing class with a physics teacher and figured that I would do sort of a biomedical engineering undergraduate degree. I mean, it would be a good way to combine my interests in human physiology and physics. But then, at that point got interested in maybe doing an MD-PhD, but didn't get into any MD-PhD programs. But yeah, I'm excited to sort of talk a little bit more about my journey to becoming a physician scientist who is studying, trying to understand why babies develop necrotizing enterocolitis. And that has sort of sparked an interest in early life immune development as well as spontaneous intestinal preparation. So thank you again for having me.

Misty Good (02:32.343)

Yeah, I'm personally so excited that you're on the podcast and we get to talk about your work, which is really transforming the field and taking a completely unbiased approach to this disease. And so, I'm just really thrilled that you're here to chat with us today. I was wondering if you could tell our listeners, I know a lot about your background and training, but maybe you could tell us or tell our listeners about your training and mentoring throughout that, and how that got you to this stage.

Bunmi Olaloye (03:09.375)

Yeah, thank you for that question. So, like I mentioned, I sort of always wanted to become a physician. And in doing an undergraduate degree in engineering, I was interested in potentially the science of medicine. But like I said, I didn't get into any PhD schools and I did my medical training at Rutgers New Jersey Medical School in Newark. And while I was there, I had the opportunity to do a sub-I in the NICU during my fourth year. And that really sort of inspired me to want to become a neonatologist. I was able to work with Dr. Fofah, who was an amazing neonatologist, who I think is still at Rutgers. And then after being in New Jersey for a few years, I wanted a change of scenery. And so, I went to my residency at the University of Texas in Galveston, where I got a really great training in sort of becoming a well-rounded neonatologist.

We had a lot of exposure to neonatology. And at the end of that, I still sort of felt like there was maybe something that I was interested in that I hadn't pursued, which was science. And so, I was lucky enough to do my fellowship at the University of Pittsburgh. And while I was there, I was like, this is my last chance to see if I could have a career in science. And I looked at a few different labs and I ended up in Dr. Liza Konikova's lab, like I mentioned earlier. And I was really sort of inspired to try to study NEC, because as we know, a lot of babies just suddenly develop NEC. I remember one particular patient during my second year of fellowship and like, I was taking care of this baby for a week or two, and everything seemed fine. And then, sort of out of the blue, the baby got really bad NEC totalis and then unfortunately did not survive. So, you know, in doing a lot of reading and research, there just really weren't a lot of answers. And I was hoping that, you know, I would get into the lab and study NEC and try to identify biomarkers and therapeutic targets to identify babies who would develop NEC and then potentially identify potential therapies, which I know Misty is also working on. But then that journey has sort of really expanded to sort of include trying to learn about how peripheral immunity develops in preterm infants, because we don't know that much about it. And then also in our initial study, we had included babies who had spontaneous intestinal perforation as like age matched controls. But it turns out that SIP also has sort of like a distinct immune landscape. And so that second piece has sort of also become a passion project as like there are even fewer studies in SIP. Yeah, that's kind of the whole story.

Betsy Crouch (06:15.426)

So many great things to talk about. I am furiously taking notes like, we have to talk about SIP versus NEC. We have to talk about, you know, I think another theme that I also want to highlight is that I also didn't get into medical school the first time I applied. I was a senior in college, and I think I applied to MD programs. And then when I didn't get in, I went to the NIH and did two years of research, and then got into MD PhD. So I do think, and yet here we are, like in a similar scenario, you know? So it's like, I think that there are many, many routes to a physician-scientist career. And, you know, I think it's about taking the best opportunity that's in front of you.

Bunmi Olaloye (06:48.161)

Right. Totally agree.

Misty Good (07:04.245)

I agree. think it's important to highlight that, right? Because, you know, so many of our listeners may be at various stages in their training, and so don't lose hope, keep going, and you never know where you'll end up. So, I think it's great to highlight.

Bunmi Olaloye (07:21.524)

Yeah, I think that I've been fortunate to have great mentors and being in places where there were a lot of great resources existing. And so that's sort of been incredibly helpful on my journey so far. And yeah, I think that, you know, to echo what you have both said, it's like, you know, if you, if at first you don't succeed, you're just like, try, try, try again.

Betsy Crouch (07:48.259)

Yeah, and you'll learn things, I think, through whatever path is in front of you. Yeah, there's a resident right now who's working in my lab, and I admire her because she came to talk to me and she said, I want to try some bench science. We have her doing a cool translational project, trying to do some work on a project that we have understanding how there are neurodevelopmental differences in kids with congenital heart disease.

So making some brain organoids in IPS lines from kids with congenital heart disease. And the reason that she said that she wanted to come be in the lab was because she said it scared her and she wanted practice overcoming the things that were scary to her. And I thought, that's a winning attitude. Yes, please come in the lab. You know?  

Misty Good (08:38.979)

Definitely, I would be like, come on in.

Betsy Crouch (08:43.086)

Isn't that great?

Bunmi Olaloye (09:06.132)

Yeah, that's, that's an amazing attitude to have. I remember sort of when I first joined the Konnikova lab, like I had done some, you know, engineering related research in undergrad, but I hadn't done any sort of wet lab research. And so, you know, I kind of showed up and I was like, yeah, like, you know, this is what I want to do. But, you know, I am willing to learn, and I did some reading, and at first I was like, well, maybe we could look in the urine, or maybe we can look in the blood. And then Liza, my mentor, was like, well, we have some samples, some intestinal tissue samples, and this lab does mass cytometry. So that's where you can start. And then I was simultaneously working on a protocol to sort of isolate immune cells from small volumes of blood. And so, both of those things have been kind of an opportunity to learn about immunology. Because for most physicians, the immunology we get in medical school is like one six week course and it's very basic. Like these are B cells, these are T cells, but being in a mucosal immunology lab was sort of an immersive learning experience and a lot of different things. And I'm still learning. So, yeah, to everyone out there who's interested in something or has a clinical question that you're interested in tackling, it's important to sort of like you can see the sort of mentors that are around and look at the resources you have and try to learn as you go.

Misty Good (10:30.467)

I think that's great. You know, we both trained at Pittsburgh. So I love, I love that connection. You know, that was me too. I didn't have any basic science experience and was learning to pipette when I was a second year fellow. So, I was right there with you. I did not learn mass cytometry at that time, but certainly that is a technique that, you know, is not new anymore, but was new at the time. 

I wonder if you could talk a little bit about learning that and the different stages and then how your science and analysis, all the different techniques that you utilize, how that's evolved since you were a fellow.

Bunmi Olaloye (11:13.912)

Yeah, in the lab was just like myself and there was a grad student and like the research tech, so it was really small and I joined. And so I think that was a great opportunity to get sort of hands on experience and all the steps that were involved. And so, you know, the lab we work in had a protocol to collect intestinal tissue samples and be able to cryopreserve them. And that was from Dr. Konnikova’s postdoc. And so I learned sort of how we got the samples. And then I was able to work through the actual steps of performing the experiment, like learning to isolate the single cells. And I think in the beginning, I showed up every day, and I was eager to learn. And at the time, I don't think I realized one, how precious the samples were, because you know, get one sample and like, you don't have an infinite number of samples or two, sort of like all the potential things that could go wrong. But we were fortunate enough to have a cohort of samples. 

So for our initial sort of mass cytometry experiment, we obviously were super interested in understanding necrotizing enterocolitis and the biobank of samples that was available in the lab. So for this experiment, we included some normal, unaffected prenatal samples that we compared to postnatal samples obtained from babies who had surgeries for necrotizing enterocolitis, patients who had surgeries for spontaneous intestinal perforation, and then patients who had surgery for congenital intestinal anomalies like intestinal atresia. And basically, for that experiment, the main goal is to first sort of take these samples and isolate the immune cells from the intestine, which we did over a one-day process. And then the next day, we take our cocktail of antibodies to identify various populations of interest. the immune cells within the intestine, including monocytes, neutrophils, T cells, B cells, natural killer cells, ILCs. And then you get an abundance of data because you have a lot of cells. And so I had the opportunity to learn how to take all that data and using sort of various pipelines available through our studio to kind of make some meaningful analysis from there. First, doing some like traditional FACS-type gating to identify which were the immune cells and then subsequently looking at various populations. And what we were able to see from there was that there's a distinct sort of immune landscape in necrotizing enterocolitis that's different from all the other groups that we compare to. And that was characterized by an influx of neutrophils and macrophages. Some of that was known from previous studies, but we were also able to sort of subtype the neutrophils. And we saw sort of three distinct populations of neutrophils that we thought corresponded to timing since the injury. And so some neutrophils were kind of more, we thought were early, like early neutrophils and then other ones were sort of like a late more age neutrophils and then some neutrophil DC like hybrids that we, that the next samples sort of fell into. And then we were able to characterize a population of monocytes that expressed some markers CD16 and CD163. CD163 is like a hemoglobin scavenger receptor that had been reported in other diseases to play both a pro and potentially anti-inflammatory role in some diseases. But we thought it was playing a pro-inflammatory role in NEC for a couple of reasons. One, this population of cells were present next to the epithelium in NEC samples. We looked at that using spatial imaging mass cytometry. Then using some single-cell RNA-seq, we were able to show that they had a pro-inflammatory signature. 

That was a crash course in translational research, going from how we get the samples, to identifying appropriate controls for an experiment, to sort of analyzing the data that we got. And that has kind of expanded into looking in the blood because we did see in some patients that we had some blood samples for this population of monocytes. And then also looking at spontaneous intestinal perforation because that had a different signature from NEC.

Betsy Crouch (18:03.448)

Mm-hmm.

Misty Good (18:28.961)

And for those SIPs, that's the focus of your K award, right?

Bunmi Olaloye (18:34.905)

Yeah, yes. so that, right. And so when I kind of saw that, like we know even less about spontaneous intestinal perforation. And we can see this sort of signature that's suggesting that there is some immunologic differences, at least at the time of injury in SIP, really suggesting that there might be some inflammation. Until now what we know of SIP is that like a small percentage of preterm infants get it. Maybe it's associated with some hypoxia, maybe it's associated with some placental abnormalities. There's some literature about relationship between indomethacin and postnatal, early postnatal steroids, but like it's really poorly understood. And so seeing some changes within the immune landscape of the intestine potentially suggest like there's something else that may be going on. And that sort of really inspired me to want to look more closely at SIP.

Misty Good (19:47.62)

That's great. It's so important because like a lot of the early NEC literature was contaminated, I think a lot by spontaneous intestinal perfs and you know, not being able to differentiate because they didn't have an exploratory laparotomy, not being able to differentiate was it NEC or SIP. You know, I think that's one of the major weaknesses of our field obviously, as you know.

Betsy Crouch (20:14.374)

I'm wondering if you could speak to this idea that our, you know, I think you're one of the people who's most able to speak to, are SIP and NEC on a continuum? Are they totally different pathophysiological processes? We were having a discussion about this in M&M the other week and I'm looking forward to seeing the data. And congratulations on a number of remarkable papers over the past couple years but you know I'm excited to see the SIP papers come out too. Come on, Bunmi, are you busy?  

Bunmi Olaloye (20:47.129)

Yeah, yeah. Yeah, yeah. I mean, I've been busy sort of trying to answer that question. And I think like Misty kind of alluded to, one of the challenges is that there is a slight overlap in presentation. Like typically we think about SIP as occurring within the first seven days or within the first 10 days of life. But there are a subset of patients that could develop NEC, and they might have a perforation, but it might end up being sort of NEC pathology. And so on a pathological level, they are sort of distinct, right? Usually, SIP is associated with, we know NEC is associated with transmural necrosis that can be sort of like diffuse, but SIP sort of histopathologically has been a little bit more vague. So historically, one thing that had been described in the literature is that it's associated with some deficiency in the muscle layer within the intestine. But that's only seen in about a third of patients. And more recently, the pathology might describe a focal injury without villus blunting. And so far, the work that we've been able to see, at least looking specifically within the immune landscape, is that in patients who develop SIP, there is a distinct signature in the T cells compared to the signature of T cells in NEC. Specifically, these T cells in SIP have a pro-inflammatory signature that's characterized by an increased expression of interferon gamma. And then they seem to have lower expression of some chemokine receptors, namely CXCR3, which I had initially thought that that meant that maybe there's some failure of migration of the T cells into the intestine that's increasing susceptibility to SIP. But we're still sort of in the process of doing some more studies to further understand why there is this signature and if it's just that SIP is affecting just the T cells or if it's sort of like more kind of global epithelial and immune cell damage. To answer your question about do they exist on a spectrum, I think that that would be like my most common hypothesis is that there's sort of like in my mind kind of like inflammatory bowel diseases of the neonate, sort of like how we conceptually think about IBD as being like Crohn's versus UC. SIP is this sort of disease that is typically localized to the terminal ileum. Meanwhile, NEC can really sort of affect anywhere in the small or large intestine.

Betsy Crouch (23:49.422)

Mm-hmm. Mm-hmm. Mm-hmm. Yeah, thank you. I'm really enjoying this because it's all of the questions that I, I don't know, that I think about. Why early? Why is one contained and the other not? Do you think, to ask a very, very simplified question is just like a way to get you to talk about other exciting things. Like, it the difference in the T cell population being affected versus neutrophils? I mean, I also, from a little bit of work we've done on germinal matrix hemorrhage and I think of neutrophils as bad actors in general. And T cell populations are a little more refined. Sometimes they're good, sometimes they're bad. Is that fair?

Bunmi Olaloye (24:47.627)

Right, yeah, I think that that's fair. I think that like, it's likely a little bit more than just the T cells. I think that, you know, potentially, you know, one, one thought that we've been sort of exploring is that as we learn more about how the mucosal immune landscape develops in humans, both in utero and in early postnatal life. We know that there's going to be sort of like their region specific differences in how the T cells and other cells develop. You know, for example, like we know that as early as 12 weeks gestation that the T cells within the intestine are predominantly memory T cells and so they have this sort of like functional phenotype and that sort of.

Betsy Crouch (25:50.786)

Wait, can we nerd out there? How do you have a memory T cell in a sterile environment in uterus?

Bunmi Olaloye (25:55.712)

Right. And so this is the other sort of like big debate is like, is the in utero environment completely sterile?

Betsy Crouch (26:03.966)

What? Wild! Yeah, okay. Tell me more.

Bunmi Olaloye (26:07.737)

I don't know. Is it completely sterile or are these memory T cells responding to metabolites, bacteria metabolites that are potentially within the amniotic fluid? Amniotic fluid swallowing begins early on and the way that the intestine development would occur would be from sort of like proximal to distal like everything else. And so, yeah, so lots of lots of questions, lots of things, lots of things to learn. But I think that the understanding of fetal mucosal immunity has sort of like expanded significantly in the past five years. And that sort of is informing how we how we think about both of these diseases, and it's kind of informing our approaches to studying them. And I mean, I think that, you know, the biggest pros are, you know, kind of access to the biobank of samples that we have and sort of collection is always ongoing and also sort of the explosion of and the development of like various single cell approaches that allows us to really get a lot of information from small tissue samples.

Misty Good (27:41.828)

I was going to say like you had mentioned earlier about indomethacin, and postnatal steroids. I'm curious, and I'm sure our listeners are curious too, like if you've been able to identify any other medications that predispose an infant to NEC or any other, I don't know, practices that we do every day in the NICU that could be predisposing our babies or setting them up.

Bunmi Olaloye (28:17.933)

Yeah, I think that that's sort of a great question. You know, in terms of NEC, as we all sort of know that like, or we think about how human milk is protective. But I think outside of that, our understanding of the pathogenesis and how various factors potentially contribute to the disease is ongoing. I mean, so far in our work, when we looked at a cohort of preterm infants, they were born at less than 30 weeks gestation, and we sort of had collected longitudinal blood samples to sort of understand how the immune system changes over time and how that's modulated by various postnatal and antenatal exposures. The only sort of link that came up from the small cohort of like 10 patients was this link that we sort of already know to like histologic chorioamnionitis. We saw that that specific exposure sort of modifies the population or alters the population of monocytes. And that change is persistent even at sort of one month of life in these samples, because we looked at blood in the first week, first month, and at two months.

And so, you know, I think it's hard to say if there are any other specific medications that change that. You know, certainly we think about how antibiotics could be potentially detrimental to the microbiome, but we haven't. Yeah, we're still sort of, still looking. These questions are tricky because NEC, as we know, is sort of heterogeneous disease and it happens, but sometimes not that often, which is good for the babies. And even when it happens, there's always sometimes a lot of confounding factors.

Misty Good (30:34.647)

Yeah, these are definitely two hard diseases to study, right? And I think, you know, for anyone out there that's thinking about doing translational research, collaboration is important, right? To be able to, you know, get the samples and be able to analyze them. Like it's really team science and all of this is a team sport. So, we're all anxiously waiting for you to tell us the answers. But you have your whole career cut out for you to figure it all out. But yeah, it's nice to hear about your K and studies related to that.

Bunmi Olaloye (31:05.977)

Yeah.

Betsy Crouch (31:14.731)

Can I nerd out for a little bit longer? I can feel Misty's getting ready to talk about the big picture and I'm like…

Bunmi Olaloye (31:20.313)

Well, now I'm wondering if you're going to ask if we thought about sort of like how the vasculature play a role since you're a vasculature biologist.

Misty Good (31:31.427)

That's exactly what you're going to ask!

Betsy Crouch (31:35.855)

No, that was question number three. There was a question one and two before that. I feel seen, thank you. That's awesome, thank you. Okay, but back to the SIP and NEC, the question I wanted to ask is about, I mean, I think it does really speak to these precious, precious human samples. And I mean, I think it is wonderful that initially that you didn't have fear when you were handling them, right? Because that's not good either. You see the person who's like shaking when they do the experiment. And I have mixed feelings because on the one hand, it is recognizing that there was a human being that contributed to the material that you're currently holding. And I don't know, sometimes it's helpful just to not think about that and just like pipette.

So the question that I wanted to ask was, animal models.  Do you feel like there's a future where we can model this in animal models? I don't know, how do you feel about Misty's neonatal mouse model? Or are you thinking about trying something different with, I mean, with a pig, a monkey? Yeah, I think that, there's there's some exciting development in translational models, like I'm collaborating on a perinatal chorio macaque project right now, because I think, you know, there is a lot of interest in trying to be thoughtful about what models we're using to answer different human specific questions.

Bunmi Olaloye (33:03.672)

Mm-hmm. Yeah, I mean, I think that that's a great question. And I think about that and how to sort of transition from where I am, right, sort of like mid-K to R and next steps. Right now, I don't have a great answer.

Misty Good (33:41.443)

It's so much.

Bunmi Olaloye (33:42.989)

I'm still sort of like still at the drawing board. But certainly, I mean, I think that you know, potentially if, you know, anyone had some ideas and was interested in collaborating, that's sort of the route to go. I think that what we've been trying to work on is potentially using organoid models. And so that's kind of the stage that we are where we're at. So there are obviously challenges with that as well.

Betsy Crouch (34:15.534)

Mm-hmm.

Misty Good (34:25.517)

Yeah, I think I was just going to say in terms of animals, like modeling SIP in a neonatal mouse, is kind of doable. It's definitely doable, but there's no good spontaneous intestinal perf. There's more or less an intestinal perf type model that you could certainly do with human enteric microbiota IP injected into a neonatal mouse, which has been done and perfected by Jim Wynn. He did an amazing job looking at using the model as a neonatal sepsis model. And so you could certainly do something like that, but the spontaneous aspect of that, it's not well understood, right - What drives the intestine to get a gut barrier breach, really, right? 

Bunmi Olaloye (35:28.789)

Right. I think that, like that, and that's sort of like the key question. And I think that's kind of like the limiting factor, at least from where SIP is concerned about sort of trying to replicate the spontaneity of it.

Misty Good (35:44.119)

Yes.

Betsy Crouch (35:46.283)

Yeah, but at least with your data set, you could sort of take a Misty Good approach, which is like take all of the things that lead to NEC, put them together in one poor neonatal mouse. And then, you know, sometimes you get NEC, right? So NEC in an animal model. And so I think it'll be exciting to see all the things that come out from, you know, your very sophisticated immune profiling in that disease, which is where I think it needs to start. Yeah, I guess another question was just which in terms of the different neutrophil populations in NEC, which one do you think is affected by breast milk? I don't know, do you have other thoughts about their heterogeneity in terms of the pathogenesis or the ability to flip it the different way? Which immune population maybe is involved with some resilience?

Bunmi Olaloye (36:48.417)

Yeah, yeah. I I think that when I think about sort of like breast milk and, you know, immune populations, at least like what we've, what's currently known is sort of how lactoferrin from breast milk modulates the MDSCs or myeloid-derived suppressant cells. And so I think that that's the population. And that, we know that in patients who have NEC, this population is sort of affected, but exposure to human milk kind of helps modulate that response. I think the other piece from sort of like breast milk and NEC protection is also sort of out of, I think he was in Pittsburgh out of Tim Hand's lab, they had been looking at sort of the impact of like maternal IgA on sort of from breast milk on NEC. And they showed that maternal IgA binding a specific bacteria could be sort of protective for NEC in some patients. And so I think that ultimately they are likely multiple components of breast milk that are modifying the immune system. The other sort of modifier that we know is that human milk derived oligosaccharides and, you know, how breast milk contains these molecules that don't get digested but are basically like food for certain types of bacteria to promote the development of certain bacterial colonies like Bifidobacteria, for example, and that piece is protective for NEC. So I think that we know kind of all these pieces. And so that's kind of interesting as well.

Betsy Crouch (38:55.554)

Yeah, thank you so much for that overview. I think this is something that when I'm rounding with the residents and I want to say like, what do we know in terms of like the molecular pathogenesis or resilience against NEC, I will refer them to your overview. So thank you. I think that was a nice educational piece too. Yeah, I mean, I won't hesitate to ask the vascular question if you have anything you have any other brilliant insights. But we don't have to go there either. I think we've had a wonderful discussion.

Bunmi Olaloye (39:34.234)

I mean, yeah, I think that we do have some, like, you know, in one of the sort of like unbiased looking at NEC using sort of single cell data, we did see that there are some, there's, you know, like, again, sort of a pro-inflammatory signature in some lymphatic endothelial cells in patients who have NEC compared to those who do not have NEC. And so, you know, I think that likely there's some, and as I mentioned earlier, the population of CD16 positive, CD163 positive monocytes that we saw spatially were in the intestine, but they were adjacent to blood vessels. And so there's likely, and if we think about how some studies have shown a drop in peripheral monocyte count right before the diagnosis of NEC, we can say potentially hypothesize that these monocytes are being recruited into the periphery through the endothelial cells and are either sort of responding to injury or are they the ones driving the injury? That sort of question is unanswered. But I do think that the endothelial cells play a role in either the development or more likely the sort of propagation of the inflammatory cascade.

Misty Good (41:09.507)

Yeah, I think that's so interesting. Like not a lot is known about the lymphatics and the association with NEC or what's going on there. So hopefully you and others can certainly share some insights there.

Bunmi Olaloye (41:26.937)

But if you want it as a vascular biologist, you want to weigh in. I mean, but then I sort of think about this. We should just do it now.

Betsy Crouch (41:32.366)

I mean, should we start sketching out the aims for our collaboration now? I mean, just to sort of think on the spot and show folks. And maybe we could submit it. I mean, yeah, please.

Bunmi Olaloye (41:47.321)

No, no, I mean, I just sort of then think about how patients who develop NEC sort of have this kind of white matter injury in response to the inflammation. And I know that, you know, David Hackam's group had really nicely shown sort of how the CD4 T cell population was sort of like pro-inflammatory and sort of migrating to the brain with some really cool mouse brain organoids. But anyway, there's clearly like a gut brain axis. And then the cellular cells are most likely.

Betsy Crouch (42:31.446)

Yeah, I mean, I think these are all just, I don't know, really enticing observations, right? Like scientifically and clinically from the samples better handle on now. Do you know Brisa Palikuqi actually? She's at Yale now.

Bunmi Olaloye (42:55.275)

Yes, yes, yes, yes, yeah, she's very interested in sort of endothelial cells and the regional specialization. We’re connected, so that's cool.

Betsy Crouch (43:05.144)

Mm-hmm. That's awesome. Yeah, I mean, she had probably the first characterization of the human intestinal lymphatic vasculature. So that's, you know, that's an awesome local person. But, yeah, I mean, I think there's, there's this outdated notion that the vasculature in terms of immune cell infiltration is like a door that's either open or shut. And I think, I mean, the paradigm I think about the most is the blood-brain barrier in regards to that paradigm. And finally, we're adopting a more nuanced understanding of like it's not open and closed doors. It's probably like, you know, many, many stages of gating in terms of, as you alluded to also, the T cell needs to adhere to the endothelium. It needs to, you know, get across to the ECM, you know, and all of these are kind of chemokine-related events also. So, when is that potentially physiologic? And when is that pathologic? I mean, the good news is they're discoverable things. We just need to keep doing the work that you're doing. Actually, all of us are doing in terms of taking the human samples, understanding the molecular players, and then really probing them in a direct

Bunmi Olaloye (44:10.457)

Mm-hmm. Right.

Betsy Crouch (44:33.07)

By the way and hopefully, you know, but could you imagine for us a future in which like we have something that prevents against SIP or we have a therapy against SIP once it happens?

Bunmi Olaloye (44:49.273)

Yeah, I mean, I think that that would be would be great. For SIP, I sort of imagine like maybe something given to sort of like higher moms that are at higher risk, just given how sort of early the incident is.

Betsy Crouch (45:07.714)

Mm-hmm. Yeah.

Misty Good (45:11.299)

I was going to zoom out little bit now, Betsy.

Betsy Crouch (45:11.608)

Yeah. Cool. Please, please, yes, as we hit the 45 minute mark. So thank you, Misty.

Misty Good (45:21.635)

I'm just curious because so many of our listeners are physician scientists and they really are probably wanting to know because you mentioned you're mid-K and so thinking about the K to R transition. As you know, this is a very vulnerable place where people sit. And I have sat there and had fear, and you had mentioned fear earlier.

Betsy Crouch (45:36.877)

Yes.

Misty Good (45:48.932)

You had mentioned it initially, like that you didn't have any fear to go into the lab. But I think the K to R transition is one that a lot of people fear. And I'm just wondering if maybe you could give some advice on, like, how you're thinking about it for your own career, and then how to help overcome that fear.

Bunmi Olaloye (46:10.891)

Yeah, no, I'm terrified of the K to R transition. You know, I think I've sort of approached my career so far, and like, you know, I’m going to take it one year at a time. And so, I have a plan for this year. And, you know, that includes, you know, taking the local grant writing course again and sort of really tapping into the resources, like mentorship and getting feedback early. And then yeah, the grant writing course here was super helpful when I first submitted my K and there were people writing ours then. And so it's just sort of getting as much feedback in the writing stage as possible, I hope it’s helpful. And then, again, like if you submit and it doesn't go all the first time, be willing to and have plans to resubmit is sort of the approach. Understanding that, I guess one of the benefits of being a physician scientist is like there is, yes, there is pressure, you know, to get that R, but I also still enjoy taking care of babies in the NICU. And so all will be well.

Misty Good (47:36.216)

That was how I got through. Yeah, that was literally how I got through. I remember thinking, okay, well, if it doesn't go well, I'll just take care of babies and be totally happy. I think that's one of the things that when you're in it, you tend to lose sigh oft. I don't know. I felt like if I put less pressure on myself to get the R or transition to independence.

You know, you mentioned transition to independence too. I think that's one of the things for me, it was one of the biggest fears, was the transition to independence and running my own lab and all of the things that go along with that. Cause it's fun to do science. And so when you're trying, you know, you're doing the experiment or doing the analysis, but then, taking that next big step where you're leading a team and inspiring the next generation and mentoring and all of the fun things that are really cool about being a PI.

Bunmi Olaloye (48:37.913)

Yeah, in the process of doing some hiring right now.

Misty Good (48:44.077)

Congrats, that's amazing. That's really great. 

Betsy Crouch (48:49.336)

Great. Yeah, thank you. We like to end on something fun about your research or about your teammates in the lab or about your colleagues in neonatology. Yeah, tell us something that you enjoy to do as a team or just something you personally enjoy when you're not pipetting or looking at the AEEG screen overnight like I was.

Bunmi Olaloye (49:22.469)

Yeah, yeah. I mean, I think when I'm not here or before I became a mom, I always sort of looked forward to playing tennis and a lot of folks in the lab play tennis. It's all indoor tennis because we're in New England. But that's something I'm hoping to sort of get back into. And now, you know, now that I have a little 10 month old at home, I basically just look forward to seeing her at the end of the day and hope she sleeps through the night.

Misty Good (49:57.441)

That is good.

Betsy Crouch (49:57.815)

Yeah. Yeah. And you, you run after her who's essentially, well, I guess not quite yet. Not yet. Yeah. How cute. 

Bunmi Olaloye (50:04.515)

Yeah, not quite yet. Not quite yet. Yeah, I can't wait. I can’t wait for the day she starts running around.  

Misty Good (50:14.285)

We also ask for pearls of advice or wisdom or anything like that for all of our early stage investigators listening.

Bunmi Olaloye (50:24.589)

Yeah, I mean, think that sort of like the biggest, you know, and I think this has come up in a few other episodes, this sort of finding good mentors who are thinking about, you know, how your work fits into their work right now, but also thinking about, you know, what's next and being very sort of helpful in connecting you with various collaborations sort of locally and internationally. You know, I think a few times I've mentioned Lisa Konikova, but she's been a great mentor. Like, she let me join her lab when I didn't know anything about immunology. And, you know, the lab had moved during the pandemic to Yale and I got an opportunity to come here to Yale as a postdoc and an attending right out of fellowship and sort of these things have been incredibly instrumental in, you know, helping to move my career forward. And so, I think that finding great mentors and just being sort of willing to learn and showing up even when you're like, I have no idea what's happening.

Misty Good (51:54.285)

Well, I think that's great but also not having any fear, right? That like you can't do it, because clearly we've all shown on this podcast, that you can do it and you've been incredibly successful. And I think for you, this is just the beginning. So keep going. And you're, really asking the questions that we all want to know at the bedside. So it's worth it. It's worth it for the babies.

Bunmi Olaloye (52:23.435)

Awesome. Yeah. Thank you so much for those kind words and for having me.

Misty Good (52:28.055)

Thank you for coming. We are just really thrilled to host you and highlight your work, and are grateful for all the work that you do.

Betsy Crouch (52:40.162)

Yeah, thank you again, Bunmi, for joining us and for enlightening us and to our listeners for engaging in our conversations as well. Well, that's it for today's episode of At the Bench, and we look forward to talking with you all soon. Take care.