Wang Yunguan, Adeleke David, Xie Xiangfei, Yang Zi, von Hofe Annika Yang, Singh Manavi, Malik Astha, Kudira Ramesh, Castro-Rojas Cyd, Pfuhler Liva, Alquraish Mosab, Sylvestre Pamela, Dillman Jonathan R, Trout Andrew T, Miraldi Emily, Miethke Alexander G
Division of Pediatric Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, 45229.
Division of Human Genetics, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, 45229.
bioRxiv. 2025 Aug 21:2025.08.14.670195. doi: 10.1101/2025.08.14.670195.
Primary sclerosing cholangitis (PSC) is an autoimmune, cholestatic liver disease characterized by inflammation and fibrosis surrounding bile ducts. The cellular crosstalk driving periductal fibrosis remains poorly defined. This study applied a multi-omics approach integrating MERSCOPE spatial transcriptomics, bulk RNA-seq, and SomaScan proteomics to characterize fibrotic periductal regions and their cell-cell communications. Macrophages (MP) expressing moderate-to-high CD163 were found co-localized with cholangiocytes, T cells, and collagen-producing hepatic stellate cells (HSC). Cell niche analysis identified periductal regions with elevated fibrotic signals, in which cell-cell communication analysis revealed MP-HSC interactions involving 17 fibrotic driver genes in MP (e.g., IFNGR1, CSF1R, CD163) and six fibrotic effector genes in HSC (e.g., COL1A2, VCAN, MMP2). In validation analyses, bulk RNA-seq data showed higher driver and effector gene scores in PSC with established fibrosis compared to early-stage PSC and autoimmune hepatitis (AIH). Plasma proteins encoded by MP driver genes were elevated in autoimmune liver disease (AILD) and in patients with elevated (≥3.29 kPa) liver stiffness on MR elastography. These findings suggest that macrophages engage in localized crosstalk with HSC, activating fibrotic gene programs and contributing to periductal fibrosis in PSC, thereby identifying potential molecular targets for therapeutic intervention.
原发性硬化性胆管炎（PSC）是一种自身免疫性胆汁淤积性肝病，其特征是胆管周围发生炎症和纤维化。驱动导管周围纤维化的细胞间相互作用仍不清楚。本研究采用多组学方法，整合MERSCOPE空间转录组学、批量RNA测序和SomaScan蛋白质组学，以表征纤维化的导管周围区域及其细胞间通讯。发现表达中到高CD163的巨噬细胞（MP）与胆管细胞、T细胞和产生胶原蛋白的肝星状细胞（HSC）共定位。细胞微环境分析确定了纤维化信号升高的导管周围区域，其中细胞间通讯分析揭示了MP-HSC相互作用，涉及MP中的17个纤维化驱动基因（如IFNGR1、CSF1R、CD163）和HSC中的6个纤维化效应基因（如COL1A2、VCAN、MMP2）。在验证分析中，批量RNA测序数据显示，与早期PSC和自身免疫性肝炎（AIH）相比，已形成纤维化的PSC中驱动基因和效应基因得分更高。MP驱动基因编码的血浆蛋白在自身免疫性肝病（AILD）和磁共振弹性成像中肝脏硬度升高（≥3.29 kPa）的患者中升高。这些发现表明，巨噬细胞与HSC进行局部相互作用，激活纤维化基因程序并导致PSC中的导管周围纤维化，从而确定了治疗干预的潜在分子靶点。
