Dr. Colleen Nofi - Best of the Best in Pediatric Surgery 2025

All right. We're gonna go on to the uh next one. this is now ABSA, and it is, uh, Colleen Nofi and it's extracellular CRP syp. What is it? SEP, uh, exacerbates necrotizing enterocolitis, and she's from Cohen Children's Medical Center in New York. Hi, my name is Colleen Nofi, and I'm a general surgery resident at Northwell Health. Thank you for the opportunity to present my work entitled Extracellular CIRP exacerbates necrotizing Intercolitis. I'm the inventor of our related patent, and I have no other conflicts of interest to report. Necrotizing entercolitis or neck is a devastating gastrointestinal disease impacting premature infants whose pathophysiology is driven by a multitude of complex pathways that are not completely understood. Further, there remains limited treatment options and an unacceptably high morbidity and mortality risk. My work focuses on a protein called cold inducible RNA binding protein, or CIRP. Under biologic conditions, CRRP is found inside the cell where it acts as an RNA chaperone protein. However, in states of cellular stress, for example, in sepsis, CIRP escapes outside the cell. Once released from the cell, CRP becomes extracellular CIRP or ECIRP, where it then acts as a damp by enhancing the release of cytokines and chemokines, and amplifying the inflammatory cascade. Previously, we developed an ECIRP scavenging peptide named MFGE8 derived oligopeptide 3, or MOP 3, that removes ECRP from circulation to reduce inflammation. The aims of my work were to determine whether ECRP plays a role in exacerbating neck pathogenesis and to evaluate the therapeutic potential of our ECRP scavenging peptide, MOP 3, to protect against neck severity and mortality. To address our aims, we utilized a mirroring model of neck where pups were separated from mothers and subjected to a multitude of stressors, including hypercaloric formula gavage, enteric LPS, and hypoxic stress. We utilized wild type and CRP knockout pups to address AIM 1, and we utilized MP3 or vehicle treatment in wild type pups to address AIM 2. First, we found that CRP knockout protected pups from neck severity. Here you can see representative H&E stained intestinal sections. Upon quantification of neck severity, we identified a reduction in neck severity scores in CRP knockout pups, which is grossly seen by the preservation of the intestinal villi architecture and shown in the green bar here. Our next experiments further evaluated the impact of CRP knockdown in the intestines. We demonstrated a reduction in intestinal inflammation in CRP knockout mice subjected to neck, as assessed by mRNA levels of IL-6 and TNF alpha in the small bowel. Here you can see that reduction represented by the green bars. To evaluate the functional impact of intestinal injury and inflammation, we utilized fluorescently labeled Dextra administered enterically during our neck model, and measured systemic fluorescence as a measure of barrier dysfunction. For example, if the intestinal barrier was compromised, the fluorescent dextrand would leak out of the injured intestinal wall and be detectable in the serum. Here you can see the greatest fluorescence intensity in wild type neck pups, indicating a leaky intestinal barrier, where CRP knockout pups had reduced fluorescence intensity, indicating a preserved intestinal barrier. Finally, to probe the potential clinical impact, we evaluated survival differences between our animal strains. Remarkably, in the same model under the same conditions, there was 100% survival for CRP knockout pups subjected to neck, whereas the survival was only 65% for wild type pups. Transitioning to our therapeutic study, we first investigated the impact of our MOP 3 treatment on systemic markers in our neck model. Here you can see that MOP 3 functioned as designed, as there was a reduction in circulating ECRP levels in MOP 3 treated neck pups compared to vehicle. This reduction in ECRP also correlated with a reduction in other systemic inflammatory markers, including IL-6 and DNF alpha. In our neck model, treatment with MOP 3 also protected against neck severity. Here in the representative H&E stained intestinal sections, you can appreciate the preservation of the intestinal villi and associated reduction in neck severity scores indicated by the orange bar. Furthermore, treatment with MOP 3 reduced intestinal inflammation in neck as measured by mRNA levels of both IL-6 and TNF alpha. Again, in probing intestinal barrier function, we observed high fluorescence in our vehicle-treated neck pups, indicating intestinal breakdown, whereas MP 3 treated pups had significantly reduced fluorescence intensity, indicating protection of the intestinal barrier. Finally, when we compared overall survival of murine pups subjected to the neck model, we found a significant improvement in survival, up to 80% in pups treated with MP3, compared to only 50% for vehicle treated pups. This further supports a beneficial role of MOP 3 treatment in experimental neck. Altogether, our data in neck suggests that ECRP exacerbates neck pathogenesis by increasing inflammation and intestinal injury, whereas treatment with our novel peptide MOP 3 protects against neck pathogenesis by scavenging ECRP and preventing the deleterious downstream impacts. Thank you to my mentors, and I'm happy to take any any questions. Perfect. Erin relations, this is really exciting and thorough work, um, and a great presentation. My question for you is, what are your next steps? You've shown really impressive, um, therapeutic benefits of mop and mice, so how do we get this to kids? So, we, uh, of course, wanna further investigate uh optimal treatment of, uh, with our peptides. So if we can give it different routes, if it can be given internally. Um, we've also been looking at, um, other models trying to expand and make sure results are reproducible since we know not a single model of neck really recapitulates everything we exhibit in, um, neonates. So right now, we're, we're already showing, um, that MOB 3 is effective in other models. of ischemia reperfusion injury in the gut. So we're trying to really understand how we can maximize the beneficial impact because you can see we have the greatest impact with CRP knockdown, whereas our MOP 3 benefit is not quite the same level. So we're just trying to maximize what we can in our um pre-clinical models right now. Can I ask you a question about your, your models? When you create neck, what is the timing of the MOP 3 administration? Is it before the neck, um, Insult or is it as, as you say, it's treatment. So is it after you create the neck? So, we actually induce a 4-day neck models. So, the stressors are actually continuous for 4 days, and they, um, consist of a multitude of factors like LPS and formula gavage and hypoxia. And so, we give the MOP 3 treatment once per day, um, at the beginning of the model. So, the treatment is, is kind of, uh, ongoing with the insultive neck. Great work. Thank you very much. Thank you. Thank you. Great job.