Assessing the Physical and Inflammatory Response of Intestinal Organoids for Necrotizing Enterocolitis using a NEC-in-a-Dish (2023)

Undergraduate: Eve Melbouci

Faculty Advisor: Misty Good
Department: Pediatrics

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Necrotizing enterocolitis (NEC) is a devastating disease that impacts the intestinal tract, which most commonly occurs in preterm infants. Its pathogenesis is unknown, but previous studies have found that an alteration in the intestinal microbiome, in particular the bacterial components and concentration, may induce the epithelial damage in NEC as well as the release of cytokines and proteins as a bodily defense. In order to assess the epithelial damage associated with NEC, a high throughput system was developed through “NEC-in-a-Dish” experiments performed on apical-out enteroids using bacteria from patients with NEC. Enteroids were dyed and exposed to the bacteria for 240 minutes and images were captured every 30 minutes. The immune response was measured using a “NEC-in-a-Dish” experiment on apical-out enteroids using bacteria from patients with NEC. Enteroids were exposed to the bacteria for 240 minutes and the RNA was run through a qPCR for quantification of TNF-α and IL-1β. Epithelial damage was observed, but there was no significant change in fluorescent signal, indicative of damage, between experimental and controls. An immune response was observed in the expression levels of TNF-α but not of IL-1β. These results will act as preliminary data for drug efficacy tests as enteroids exposed to NEC bacteria are treated with possible drugs.

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