Chen Wenyi, Chen Xinyi, Gao Feiqiong, Yao Qigu, Cheng Sheng, Pan Qiaoling, Yu Jiong, Yang Jinfeng, Ma Guanghua, Gong Jintao, Li Qian, Chen Yunhua, Lim Lee Wei, Stambler Ilia, Ellison-Hughes Georgina M, Ulfhake Brun, Zhao Robert Chunhua, Cao Hongcui
State Key Laboratory for the Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, National Medical Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, 79 Qingchun Rd, Hangzhou, 310003, China.
Department of Endoscopy, Zhejiang Cancer Hospital, Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences, Hangzhou, Zhejiang, 310022, China.
J Nanobiotechnology. 2025 Jul 28;23(1):546. doi: 10.1186/s12951-025-03617-2.
Primary sclerosing cholangitis (PSC) pathogenesis involves immune dysregulation, genetic factors, and bile duct pathology; however, a comprehensive pathogenesis model and effective therapeutic strategies remain limited. Here, we develop a novel human liver multilineage organoid (Mulorg) model combined with Mdr2 mice to investigate the pro-fibrotic role of T helper 17 cells (Th17) and the therapeutic potential of mesenchymal stem cell-derived extracellular vesicles (EV) for PSC, particularly periductal fibrosis. EV alleviates interleukin-17A (IL-17A)-induced fibrotic Mulorgs (FibHOs) and mitigates periductal fibrosis in Mdr2 mice by inhibiting Th17 differentiation, decreasing Th17 numbers, and lowering intrahepatic IL-17A levels. Functional assays, miRNA array, and CUT & Tag analyses reveal that EVs-derived hsa-miR-7977 targets NFKBIZ, repressing IκBζ translation to reduce IL-17A and its downstream targets involved in Th17 differentiation, IL-17 signaling, and bile secretion pathways. Moreover, miR-7977-enriched EV efficiently reduces IL-17A cell percentages in fibrotic areas and improves periductal fibrosis in Mdr2 mice. Co-culture of FibHOs with Th17 found miR-7977 inhibits Th17 migration to the periductal fibrosis area, with distinct morphological differences observed between patient- and healthy-derived FibHOs. These findings demonstrate that EV-derived miR-7977 mitigates the periductal fibrosis microenvironment by inhibiting Th17 differentiation and migration, the former by targeting NFKBIZ, regulating IL-17A and IκBζ-targeted gene expression. This study clarifies Th17's role in the PSC fibrotic microenvironment, underscores the modeling contributions of Mulorgs, and highlights EV-derived miR-7977's potential to ameliorate Th17-related periductal fibrosis, offering insights and novel therapeutic avenues for PSC.
原发性硬化性胆管炎（PSC）的发病机制涉及免疫失调、遗传因素和胆管病理变化；然而，全面的发病机制模型和有效的治疗策略仍然有限。在此，我们开发了一种新型的人肝多谱系类器官（Mulorg）模型，并结合Mdr2小鼠，以研究辅助性T细胞17（Th17）的促纤维化作用以及间充质干细胞衍生的细胞外囊泡（EV）对PSC，特别是导管周围纤维化的治疗潜力。EV通过抑制Th17分化、减少Th17数量和降低肝内白细胞介素-17A（IL-17A）水平，减轻白细胞介素-17A（IL-17A）诱导的纤维化Mulorg（FibHOs），并减轻Mdr2小鼠的导管周围纤维化。功能测定、miRNA阵列和CUT&Tag分析表明，EV衍生的hsa-miR-7977靶向NFKBIZ，抑制IκBζ翻译，以减少IL-17A及其参与Th17分化、IL-17信号传导和胆汁分泌途径的下游靶点。此外，富含miR-7977的EV有效降低纤维化区域中IL-17A细胞百分比，并改善Mdr2小鼠的导管周围纤维化。FibHOs与Th17共培养发现，miR-7977抑制Th17向导管周围纤维化区域的迁移，在患者来源和健康来源的FibHOs之间观察到明显的形态差异。这些发现表明，EV衍生的miR-7977通过抑制Th17分化和迁移减轻导管周围纤维化微环境，前者通过靶向NFKBIZ，调节IL-17A和IκBζ靶向的基因表达。本研究阐明了Th17在PSC纤维化微环境中的作用，强调了Mulorgs的建模贡献，并突出了EV衍生的miR-7977改善Th17相关导管周围纤维化的潜力，为PSC提供了见解和新的治疗途径。
