肝脏巨噬细胞分子图谱及调控乙肝重症化机制研究

浙江省乙肝发病率仍处于全国较高水平，乙肝重症化相关并发症死亡率高。肝脏巨噬细胞为肝内主要免疫细胞群，在乙肝的发生、发展和转归中起重要作用，但肝脏巨噬细胞在乙肝重症化中的作用呈高度异质性，传统的M1和M2亚群分型方法无法精确认识其在乙肝重症化中功能表型变化。单细胞RNA-Seq技术可揭示每个细胞独特微妙变化，我们利用经典单细胞测序和自主研制的单细胞小RNA和mRNA双测技术进行肝脏免疫细胞分型研究，初步发现Trem2、Emp1等分子标记可区分不同功能的疾病相关巨噬细胞亚群。本项目拟通过对乙肝临床病例重症化进程中肝脏微环境内巨噬细胞亚群的解析，经乙肝动物模型及基因靶向干预手段进一步识别调控其功能演变的关键超级增强子、小RNA与基因等，同时解析巨噬细胞与肝内其他细胞的互作网络，并在乙肝临床队列加以验证。有望实现乙肝肝脏巨噬细胞的精准分型及亚群功能鉴定，为阐明乙肝重症化的分子免疫机制提供新思路。

The incidence of hepatitis B in Zhejiang Province is still at a relatively high level in the country, and the mortality rate of complications associated with execerbation of hepatitis B is high. Liver macrophages, as the main immune cell population in the liver, play an important role in the occurrence, development and outcome of hepatitis B, but the role of liver macrophages in the execerbation of hepatitis B is highly heterogeneous. The traditional classification method cannot accurately recognize its functional phenotype changes in the execerbation of hepatitis B. Single-cell RNA-Seq technology can reveal the unique and subtle changes of each cell. We used classic single-cell sequencing and self-developed single-cell small RNA and mRNA dual-test technology to conduct liver immune cell typing research and initially found molecular markers such as Trem2 and Emp1, which can distinguish subpopulations of disease-related macrophages with different functions. This project intends to further identify key super-enhancers, small RNAs and genes that regulate the evolution of their functions through the analysis of macrophage subpopulations in the liver microenvironment during the execerbation of hepatitis B in clinical cases, simultaneously analyzing the interaction network between macrophages and other cells in the liver. Both of them will be verified in the hepatitis B clinical cohort. Our project is expected to achieve accurate molecular typing and subgroup functional identification of liver macrophages in hepatitis B, and provide new ideas for elucidating the molecular immune mechanism of the execerbation of hepatitis B.