Pediatric intestinal enteroids derived from very early onset inflammatory bowel disease reveal differences in growth, morphology, and barrier function

Pediatric inflammatory bowel disease (IBD) presents more aggressively than adult-onset disease; however, epithelial pathophysiology remains poorly understood due to limited access to patient tissue. Human intestinal enteroids (HIEs) are a powerful model for investigating disease phenotypes and heterogeneity. We characterised duodenal HIEs from two cases with Crohn’s disease (ED-71 and ED-81) and one control with intestinal obstruction (ED-19), all aged <2years. Analyses included growth curves, epithelial cell heights, immunofluorescence, histology, electron microscopy, gene expression and intestinal permeability. All HIEs maintained consistent growth in vitro, with ED-81 displaying the steepest trajectory. Morphological examination revealed variations in epithelial cell height, with ED-71 and ED-81[median(95%CI): 10.2(7.76–11.4) and 12.5(11.5–15)µm] displaying a more flattened appearance than ED-19[18.7(16.8–21.0)µm, p-value:<0.001]. HIE differentiation into intestinal cell types was confirmed by gene expression and microscopy. Intestinal permeability assays indicated compromised barrier integrity in IBD-derived monolayers, with ED-81 exhibiting the highest baseline permeability (5.27%) and EGTA-induced disruption (50.03%) compared to ED-19 (3.88% and 36.82%). Pediatric intestinal enteroids in IBD demonstrate differences in epithelial growth, morphology, and barrier function. HIEs serve as a potential translational model for pediatric IBD, facilitating the study of epithelial pathophysiology and guiding precision therapy. Further studies with more pediatric HIEs are needed to confirm these findings.