胆道类器官揭示胆道闭锁中延迟的上皮发育和屏障功能

Background: RNASeq was performed on organoids derived from livers of normal healthy donors and patients with biliary atresia to characterize transcriptomic signatures. Methods: Organoids generated from livers of normal healthy donors and patients with biliary atresia were cultured either in expansion (undifferentiated: 3 NCOs and 11 BACOs) or differentiation medium (differentiated: 3 BACOs). Liver tissues obtained from deceased-donor subjects served as normal controls (N=3). Total RNA was isolated from organoids and liver biopsy tissue specimens. Results: Organoids from patients with biliary atresia showed abnormal cell polarity, loss of tight junctions, increased permeability and decreased expression of genes related to epidermal growth factor (EGF)- and fibroblast growth factor 2 (FGF2)-signaling. When treated with EGF+FGF2, biliary atresia organoids expressed differentiation and functional markers with restored cell polarity. Conclusion: Organoids from biliary atresia are viable and have evidence of halted epithelial development. The induction of developmental markers, improved cell‐cell junction, and decreased epithelial permeability by EGF and FGF2 identifies potential strategies to promote epithelial maturation and function.背景：本研究对来自健康供体肝脏及胆道闭锁（biliary atresia）患者肝脏的类器官（organoids）开展转录组测序（RNASeq），以解析其转录组特征。 方法：从健康供体及胆道闭锁患者肝脏中分离得到的类器官，分别于扩增培养基（未分化状态：3个NCOs与11个BACOs）或分化培养基（分化状态：3个BACOs）中培养。以已故供体获取的肝脏组织作为正常对照（N=3）。从类器官及肝活检组织标本中提取总RNA。 结果：胆道闭锁患者来源的类器官表现出细胞极性异常、紧密连接缺失、通透性升高，且与表皮生长因子（EGF）及成纤维细胞生长因子2（FGF2）信号通路相关的基因表达水平下调。当用EGF+FGF2处理后，胆道闭锁类器官可表达分化相关及功能相关标志物，且细胞极性得以恢复。 结论：胆道闭锁来源的类器官具有存活能力，且存在上皮发育停滞的特征。EGF与FGF2可诱导发育标志物表达、改善细胞间连接并降低上皮通透性，这为促进上皮成熟与功能恢复提供了潜在干预策略。

搜集汇总

背景与挑战

背景概述

该数据集（GSE186444）是一个转录组学数据集，使用RNA-Seq技术对来自正常健康供体和胆道闭锁患者的肝源性类器官进行分析，包含20个样本，数据总量为42.7 GB。研究揭示了胆道闭锁类器官存在上皮发育延迟和屏障功能缺陷，表现为细胞极性异常、紧密连接丧失及EGF/FGF2信号通路基因表达减少，而EGF和FGF2处理能恢复分化标记和细胞功能，为促进上皮成熟提供了潜在策略。