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. To investigate MSCs-derived extracellular vesicles (EVs) on preterm intestinal epithelial maturation, the preterm intestinal organoids (GA27) were treated with EVs derived from two distinct lines of MSCs, designated EV39 and EV42, for 7days and subjected to transcriptomic profiling via RNA-seq.

早产会破坏肠道上皮细胞的关键成熟进程，进而导致消化与吸收功能受损。本研究探讨了间充质干细胞（mesenchymal stem cell, MSC）来源的细胞外囊泡（extracellular vesicles, EVs）促进早产人类肠道上皮细胞成熟的潜力。
本研究通过构建源自孕27周新生儿回肠组织的早产人类肠道类器官，证实EVs（尤其是EV39株）处理可显著上调成年型肠上皮细胞标志物的表达，这些标志物涵盖参与脂质与营养物质转运、消化酶合成以及上皮屏障完整性维持的相关基因。
我们通过对多组单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）数据集进行重新分析，鉴定出上述成熟特征，揭示了独特的肠上皮细胞分化轨迹以及成熟特异性的关键基因表达谱。
值得注意的是，这些成熟促进效应依赖于囊性纤维化跨膜传导调节因子（cystic fibrosis transmembrane conductance regulator, CFTR）的活性，抑制CFTR可逆转上述有益作用。
蛋白质组学分析鉴定出成纤维细胞生长因子2（FGF2）与转化生长因子β1（TGFβ1）是介导EV39发挥作用的关键配体。
联合使用FGF2与TGFβ1可进一步提升上皮屏障完整性与脂肪酸摄取能力，这凸显了EV39或其配体在促进早产婴儿肠道功能成熟方面的转化应用潜力。
为探究MSC来源的EVs对早产肠道上皮成熟的影响，本研究将早产肠道类器官（GA27）用两种不同MSC系来源的EVs（分别命名为EV39与EV42）处理7天，随后通过RNA测序开展转录组谱分析。