Organ Transplant Center, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Guangdong Provincial Key Laboratory of Organ Donation and Transplant Immunology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Front Immunol. 2023 Feb 9;14:1034356. doi: 10.3389/fimmu.2023.1034356. eCollection 2023.
The risk of alcoholic cirrhosis increases in a dose- and time-dependent manner with alcohol consumption and ethanol metabolism in the liver. Currently, no effective antifibrotic therapies are available. We aimed to obtain a better understanding of the cellular and molecular mechanisms involved in the pathogenesis of liver cirrhosis.
We performed single-cell RNA-sequencing to analyze immune cells from the liver tissue and peripheral blood form patients with alcoholic cirrhosis and healthy controls to profile the transcriptomes of more than 100,000 single human cells and yield molecular definitions for non-parenchymal cell types. In addition, we performed single-cell RNA-sequencing analysis to reveal the immune microenvironment related to alcoholic liver cirrhosis. Hematoxylin and eosin, Immunofluorescence staining and Flow cytometric analysis were employed to study the difference between tissues and cells with or without alcoholic cirrhosis.
We identified a fibrosis-associated M1 subpopulation of macrophages that expands in liver fibrosis, differentiates from circulating monocytes, and is pro-fibrogenic. We also define mucosal-associated invariant T (MAIT) cells that expand in alcoholic cirrhosis and are topographically restricted to the fibrotic niche. Multilineage modeling of ligand and receptor interactions between the fibrosis-associated macrophages, MAIT, and NK cells revealed the intra-fibrotic activity of several pro-fibrogenic pathways, including responses to cytokines and antigen processing and presentation, natural killer cell-mediated cytotoxicity, cell adhesion molecules, Th1/Th2/Th17 cell differentiation, IL-17 signaling pathway, and Toll-like receptor signaling pathway.
Our work dissects unanticipated aspects of the cellular and molecular basis of human organ alcoholic fibrosis at the single-cell level and provides a conceptual framework for the discovery of rational therapeutic targets in liver alcoholic cirrhosis.
酒精性肝硬化的风险与酒精摄入和肝脏中的乙醇代谢呈剂量和时间依赖性增加。目前，尚无有效的抗纤维化治疗方法。我们旨在更好地了解肝纤维化发病机制中涉及的细胞和分子机制。
我们进行了单细胞 RNA 测序，以分析来自酒精性肝硬化患者和健康对照者的肝组织和外周血中的免疫细胞，以分析超过 100,000 个人类单细胞的转录组，并为非实质细胞类型提供分子定义。此外，我们进行了单细胞 RNA 测序分析，以揭示与酒精性肝硬化相关的免疫微环境。苏木精和伊红、免疫荧光染色和流式细胞术分析用于研究有或没有酒精性肝硬化的组织和细胞之间的差异。
我们鉴定了一种纤维化相关的 M1 巨噬细胞亚群，该亚群在肝纤维化中扩张，从循环单核细胞分化而来，并具有促纤维化作用。我们还定义了在酒精性肝硬化中扩张且在纤维化部位局限的粘膜相关不变 T (MAIT) 细胞。纤维化相关巨噬细胞、MAIT 和 NK 细胞之间配体和受体相互作用的多谱系建模揭示了几个促纤维化途径的纤维化内活性，包括对细胞因子和抗原加工和呈递、自然杀伤细胞介导的细胞毒性、细胞粘附分子、Th1/Th2/Th17 细胞分化、IL-17 信号通路和 Toll 样受体信号通路的反应。
我们的工作在单细胞水平上剖析了人类器官酒精性纤维化的细胞和分子基础的意外方面，并为在酒精性肝硬化中发现合理的治疗靶点提供了概念框架。
