乙肝恶性转化过程中病毒变异与炎症网络的交互作用机制

Hepatitis B virus (HBV) mutations and inflammation-regulating networks have been significantly associated with the development of hepatocellular carcinoma (HCC), respectively. However, their synergistic role on the promotion of HCC remains largely unknown. In this project, we aim to explore the synergistic roles of the HBV mutations and proinflammatory molecular networks on the promotion of HCC via integrating the findings of three previous projects with complementarity and the networks regulated by somatic “driving” mutations in HCC. We will carry out the following investigations in this project: 1. To clarify the roles of the HBV mutations and key signaling pathways regulated by deep sequencing-derived somatic “driving” mutations on the formation of heterogenicity in postoperative HCC recurrence, replenish and verify the roles of 4 key inflammatory signaling pathways identified in the 3 previous studies. 2. To elucidate the the interactions of inflammatory network nodule factors with the HBV mutations as well as their associations with HCC occurrence and recurrence. 3. To determine the etiological roles of previously identified signaling pathways, somatic mutations-regulated novel pathway molecules, and the HBV mutations on HCC occurrence and postoperative recurrence. 4. To determine the effect of HBV mutations on the regulation of proinflammatory signaling networks, the effect of proinflammatory signaling networks on the replication of HBV, and both in promoting the development of HCC and the mechanisms in cell models, animal models, and patient-derived xenograft models, consecutively. This integrative project will of great significance in elucidating the mechanism of HBV-induced hepatocarcinogenesis, enriching the theory of Cancer Evo-Dev, identifying novel surveillance and prognostic biomarkers as well as therapeutic targets to improve prediction, active prophylaxis, and targeted treatment for HBV-related HCC.

乙肝病毒（HBV）变异和炎症调控网络与肝细胞癌（HCC）有显著关联，但尚不清楚两者协同促癌机制。本课题拟整合三个互补性课题的前期发现，结合HCC体细胞驱动变异调控网络，探索HBV变异和炎症调控网络协同促进HCC进化发育的分子机制。拟采用深度测序阐明体细胞驱动变异调控的关键信号通路与HBV变异在HCC再发异质性形成中的作用，补充和验证前期发现的四个主要信号通路；阐明炎症网络节点分子与HBV变异的交互作用与HCC发生和再发的关联性；确定四个主要信号通路和体细胞变异调控的新通路分子与HBV变异对HCC发生和预后的贡献；通过细胞、动物和HCC组织移植模型，阐明HBV变异对炎症信号网络的调控作用和炎症信号网络对HBV复制的调控作用，以及二者对HBV-HCC的促进作用及其分子机制。本课题对阐明HBV致癌机制，完善癌症进化发育学理论，发现HCC发生和预后标志及治疗靶标，对有效预防和治疗HCC有重要意义。

本项目旨在阐明乙肝病毒（HBV）变异和炎症调控网络在肝细胞癌（HCC）进化中的交互作用。利用测序分析、细胞和动物模型实验、大规模病例-对照研究以及队列研究，阐明了炎症免疫促进病毒和体细胞突变产生、病毒感染和突变反作用于炎症环境、以及二者协同促进HCC进化的机制，确定了相关节点分子的应用价值。.证实HBV整合引发的高频体细胞突变与细胞无限增殖以及能量代谢异常等“干性”特征有关；炎症相关的致突变基因表达与转录组的HBV整合水平呈正相关；癌旁组织中HBV整合水平过高提示预后不良。确证炎症因子IL-6在上调促突变分子APOBEC3B表达的同时可以抑制DNA修复分子UNG的表达，造成基因损伤-修复失衡。通过5221例的病例-对照研究证实，促进APOBEC3B-UNG失衡的遗传多态性可以增加HBV突变频率和HCC发生风险。HBV感染者队列研究证明，在未接受抗病毒治疗的HBV慢性感染者中，HBV PreS缺失无促癌作用而PreS区的突变组合（G2950A/G2951A/A2962G/C2964A）可以增加HCC发生风险；在接受抗病毒治疗的HBV慢性感染者中，PreS区缺失突变可以增加癌症风险而突变组合无促癌作用。在体内和体外研究中证实，HBV S区和X区的HCC相关突变可以分别促进STAT3通路激活以及改变细胞周期调控蛋白表达，进而增强细胞恶性表型，抑制HBV突变致癌的信号通路节点分子CDC20、PAI1可以阻滞HCC增殖。抑癌基因ZHX3可以通过降低HBV启动子活性和改变cccDNA表观遗传修饰的功能抑制HBV复制，而HBx可以激活miR-155进而下调ZHX2的表达水平并促进癌症发展。HBV慢性感染可以通过上调Tim-3抑制自然杀伤细胞功能、激活NF-κB/IL-6/STAT3炎症通路，进而促进HCC进化。体外研究证实，肝癌细胞分泌的单核/巨噬细胞趋化因子（CCL2）可以抑制抗肿瘤免疫反应，而靶向抑制CCL2受体的小分子拮抗剂（747）与索拉非尼联合使用可以显著提高抗肿瘤效果。.本课题深入阐明了炎症环境、HBV变异、免疫反应以及体细胞突变在HCC进化过程中的紧密关系，完善了癌症进化发育理论；筛选了新的标志物并验证了其在HCC发生风险评估、预后预测以及靶向治疗中的价值，为进一步提高HBV-HCC的三级预防水平奠定了基础。