Warburg 效应与HBV感染慢性炎症恶性转化的分子机制

项目名称： Warburg 效应与HBV感染慢性炎症恶性转化的分子机制

项目编号： No.91529103

项目类型： 重大研究计划

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 廖勇

作者单位： 重庆医科大学

项目金额： 120万元

中文摘要： 在前期研究中，通过对感染HBV病人肝穿样本与正常肝组织的差异蛋白组学检测，发现在HBV感染肝细胞参与调控有氧酵解（即Warburg效应）的代谢酶谱的上调表达变化，并且，通过体内、外的实验，初步验证了HBV的感染、复制与抗病毒治疗反应，也与Warburg效应机制有关联。因此，我们的工作假说是： Warburg效应是细胞为适应和满足HBV病毒复制的特殊代谢需求而启动，在慢性炎症过程中，长期性的HBV病毒复制与宿主细胞对代谢与氧化应急的反复适应的结果，必然引起细胞代谢程序的重组（metabolic reprogramming），而代谢程序的重组则正是“炎－癌”转化的主要诱因和驱动因素。因此， 我们设计了从慢性复制性HBV稳定感染细胞株、“炎－癌”进展不同临床阶段的病人样本、以及不同免疫力与特定遗传修饰背景小鼠HBV感染模型等三个层次的实验，利用现代“组学” 技术，结合生物信息学方法来验证该假说。

中文关键词： C09_肝和肝内胆管肿瘤；慢性病毒性肝炎；代谢程序重组；代谢组学；Warburg；效应

英文摘要： Cancer cells are well documented to rewire their metabolism and energy production networks via aerobic glycolysis, or the Warburg effect, to support and enable rapid proliferation, continuous growth, survival in harsh conditions, invasion and metastasis, and resistance to anti-cancer drugs. During chronic viral infection, replication of the viral genome and packaging and secreting viruses may request the host cells to employ a similar Warburg-like metabolic program and energy production networks operated efficiently in the proliferating cancer cells, in order to provide the high energy needs and building blocks for macromolecular synthesis, such as the synthesis of nucleotides, lipids, and proteins. Although few report documents the involvement of Warburg effect in HBV replication and during the processes of HBV-associated hepatocarcinogenesis, our preliminary studies demonstrate that several proteins that critically control the metabolic flux during glycolysis are deregulated in the replicative-competent HBV stable cells and a hepatocellular cancer (HCC) cell line derived from a HBV-positive patients by an unbiased iTRAQ proteomic analysis. In addition, we showed that the signaling pathway that controls the Warburg glycolytic metabolism of glucose is also involved in HBV infection, viral replication, as well as the clearance of HBV virus by anti-viral drugs. Therefore, it is our central hypothesis that the metabolic adaptation and metabolic reprogramming during chronic inflammation mediated by HBV infection and viral replication is the major mechanism underlying HBV-associated hepatocarcinogenesis and (epi-)genetic alterations that initiate a Warburg-like metabolic reprograming in the HBV-replicating host cells is the critical step toward the malignant transformation. To test this hypothesis, we plan to investigate the cellular intrinsic networks that control the Warburg effect in the replicative HBV-transfected stable cells in the presence or absence of inhibitors that block metabolic flux during glycolysis at various levels; blood and tissue samples obtained from patients with HBV-associated hepatitis, fibrosis, cirrhosis, and HCC; and animal models with chronic HBV-infection in immune competent or immune incompetent mice, and with or without initiation by a chemical carcinogen diethylnitrosamine in a diverse genetic background via the CRISPR-CAS9-mediated genome editing. We expect that data obtained from these experiments, after bio-computational analysis, will help us to unveil how the metabolic program goes awry during the processes from chronic inflammation, initiated by HBV infection, to malignant transformation. We also expect to identify several key players involved in triggering Warburg-like metabolic reprogramming in the host cells with HBV infection during the malignant transition, which may serve as potential biomarkers for early screening of HCC and as interventional targets for prevention of malignant transformation.

英文关键词： Hepatocellular carcinoma;Chronic viral heptitis ;metabolic reprogramming;metabolomes;Warburg effect