基于单细胞多维组学研究单癌基因启动的肝癌的细胞异质性和调节机制

肝癌（指肝细胞癌）的肝内异质性认识不足严重阻碍了肝癌患者的分子分型、诊断和治疗。我们前期建立了一种c-Myc特异诱导肝癌的遗传修饰小鼠模型并显示瘤结节间转录组和突变谱的高度异质性，在此提出：单癌基因启动肝癌产生多组学及表型的细胞异质性；微环境和表观调控对此演化发挥重要作用，塑造原始克隆的命运。我们将深入刻画上述模型的肝癌细胞时空病理特征，并以结节为单位实施多阶段的单细胞测序，阐明基因突变、表达、DNA甲基化及开放染色质等多维度整合的癌细胞分层及多组学互作；采用空间转录组技术剖析肝微环境对癌细胞异质性的调控；分离重要癌细胞亚群并通过离体和同系小鼠移植验证其肿瘤生物学行为；比较患者肝癌细胞异质性且鉴定有临床价值的分子标记。本研究旨在从单基因启动肝癌原生模型入手解析肝癌细胞异质性的起源和演化规律、表观修饰及微环境调控机制，探讨肝癌细胞的组学和表型命运，促进肝癌分子机理的认识及精准医疗。

The poor understanding of intrahepatic heterogeneity of hepatocellular carcinoma (HCC) severely hinders its molecular classification, diagnosis and treatment. We recently generated a genetically modified mouse model of c-Myc-induced multinodular HCC, which showed high heterogeneity of transcriptome and mutation spectrum between the nodes. We here propose that c-Myc alone can initiate HCC with highly cellular heterogeneity at levels of multi-omics and phenotype, and that microenvironment and epigenetic regulation play an important role in the evolution and shape the fate of an original HCC clone. We will characterize the spatiotemporal pathological heterogeneity of HCC in the model, and carry out single cell multiomics sequencing for each node at multi-stages, so as to clarify the stratification of HCC cells with multi-dimensional integration, and the interaction among gene mutation, expression, DNA methylation and open chromatins. We will also with spatial transcriptome technology dissect the microenvironmen regulation for HCC heterogeneity. We will isolate significant subpopulations of HCC cells, and study their biological characteristics by in vitro test and isogeneic mice transplantation. Finally we will compare the result achieved with HCC patients and validate a set of molecular markers of clinical potential. The purpose of this study is to reveal the origin and evolution of HCC heterogeneity and its regulation with epigenetic modification and microenvironment from the perspective of single gene-induced autochthonous HCC model, and to explore the fate of HCC cells on the level of multiomics and phenotype. The output of this study will promote the understanding of the molecular mechanism of HCC heterogeneity, and speed up precise medicine for HCC.