Targeting CFTR and FGF2 and TGFβ1 Pathways with MSC-Derived Extracellular Vesicles to Promote Functional Maturation of Preterm Intestinal Epithelium

Preterm birth disrupts the critical maturation of the intestinal epithelium, leading to compromised digestive and absorptive functions. This study explores the potential of mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) to promote the maturation of human preterm intestinal epithelial cells. By establishing human preterm intestinal organoids derived from the ileal tissue of a neonate born at 27 weeks gestational age, we demonstrate that treatment with EVs, particularly the EV39 line, significantly enhances the expression of adult-type enterocyte markers, including genes involved in lipid and nutrient transport, digestive enzymes, and epithelial barrier integrity. These maturation signatures were identified through reanalysis of multiple single-cell RNA sequencing (scRNA-seq) datasets, revealing distinct enterocyte differentiation trajectories and key maturation-specific gene expression profiles. Notably, these maturation effects were dependent on cystic fibrosis transmembrane conductance regulator (CFTR) activity, as CFTR inhibition reversed the observed benefits. Proteomic analysis identified FGF2 and TGFβ1 as key ligands mediating the effects of EV39. Co-treatment with FGF2 and TGFβ1 further enhanced epithelial barrier integrity and fatty acid uptake, highlighting the translational potential of EV39 or its ligands in promoting intestinal functional maturation in preterm infants.