HCV Strain Variation and HCC

HCV 毒株变异与 HCC

• 批准号: 8115638
• 负责人: Andrea D. Branch
• 金额: $ 22.12万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-09 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8115638
• 关键词: Antibodies; Binding; Biological; Biological Markers; C-terminal; Cancer Etiology; Cell Culture Techniques; Cells; Chronic Hepatitis C; Cirrhosis; Clinical; Clinical Research; Codon Nucleotides; Core Protein; Data; Development; Diagnostic tests; Event; Exposure to; Family; Future; Gene Expression; Gene Mutation; Genes; Goals; Hepatitis C Antiviral; Hepatitis C virus; Hepatocarcinogenesis; Hepatocyte; Individual; Insulin Resistance; Interferons; Investigation; Japan; Kinetics; Lead; Life Cycle Stages; Liver; Logistic Regressions; Luciferases; Malignant Neoplasms; Malignant neoplasm of liver; Measures; Methods; Molecular; Mutation; N-terminal; Nested Case-Control Study; Odds Ratio; Oncogenic; Outcome; Patients; Point Mutation; Population; Primary carcinoma of the liver cells; Process; Protein Biosynthesis; Proteins; Publishing; RNA replication; Reporter; Research Personnel; Resistance; Risk; Signal Pathway; Specimen; Susceptibility Gene; Testing; Therapeutic Intervention; Time; Treatment Failure; United States; Variant; Viral; Viral Physiology; Viral Proteins; Virulence; Virulent; Virus; Western Blotting; Western Europe; Work; base; carcinogenesis; clinically relevant; fitness; in vivo; insight; member; metaplastic cell transformation; mortality; primary outcome; protein expression; research study; response; secondary outcome; tumor; tumorigenesis; viral RNA; virus core

DESCRIPTION (provided by applicant): Hepatocellular carcinoma (HCC) is the third most common cause of cancer-related mortality in the world. HCV is a major cause of HCC in the United States, Western Europe, and Japan and therefore it is critical to determine the molecular basis of the relationship between HCV and HCC. Published data show that viral point mutations in codons 70 and 91 of the HCV core gene are associated with the development of HCC, interferon (IFN) treatment failure, and insulin resistance. These viral mutations provide a compelling starting point for mechanistic studies of HCV-related hepatocellular carcinogenesis. We recently discovered that these mutations dramatically alter HCV gene expression and control the levels of two previously-unknown HCV proteins, 70 and 91 minicore proteins (Eng et al, 2009, J Virol). The proposed experiments examine the impact of these mutations, and two additional HCC-related HCV mutations that we identified (Fishman et al, 2009, Clin Cancer Res). The goal of Aim I is to determine whether core gene mutations arise prior to the development of liver cancer and thus could potentially promote hepatocellular carcinogenesis. A nested case-control study will be performed to compare core gene sequences of "cases" who developed HCC to those of "controls" who did not develop HCC during the Hepatitis C Antiviral Long-Term Treatment Against Cirrhosis (HALT-C) Trial. The primary outcome will be the percentage of cases versus controls in which the majority of the variants in the viral RNA population (i.e., the quasispecies) has a codon 70 mutation. Multivariable analysis will be performed to determine whether the codon 70 mutation is an independent predictor of incident HCC. Secondary outcomes will include associations between mutations in other codons and clinical outcomes, and changes in the quasispecies over time. The experimental goal of Aim II is to determine whether the HCC-related mutations confer a survival advantage to HCV during exposure to IFN in cell culture. Viral RNA and protein kinetics will be measured over time by qRT/PCR and Western blotting, using our established methods. These studies will determine the impact of four HCC-related mutations on HCV RNA replica- tion, protein synthesis (including minicore synthesis), and on the ability of HCV to withstand IFN treatment. The in vivo expression of minicores will be confirmed through analysis of liver specimens. This project challenges the paradigm, widely-held by investigators in the United States, that all HCV variants have an equivalent oncogenic potential. Accomplishment of our Aims will fill fundamental gaps in understand- ing the molecular basis of HCV-related oncogenesis and may lead to the identification of new targets for therapeutic interventions and to the development of diagnostic tests for patients harboring oncogenic variants. PUBLIC HEALTH RELEVANCE: The hepatitis C virus (HCV) is a major cause of hepatocellular carcinoma (HCC), the third most common cause of cancer-related mortality in the world. Clinical studies indicate that point mutations in the HCV core gene enhance HCC risk and increase the likelihood of interferon treatment failure. Because of their clinical relevance, we will define the molecular effects of four HCC-associated mutations on viral replication, and also determine whether these mutations arise prior to HCC and therefore warrant investigation as possible biomarkers of virulent strains that can be used to identify patients who have an elevated risk of developing liver cancer.

描述（由申请人提供）：肝细胞癌（HCC）是世界上与癌症有关的死亡率的第三大原因。 HCV是美国，西欧和日本HCC的主要原因，因此确定HCV与HCC之间关系的分子基础至关重要。发表的数据表明，HCV核心基因的密码子70和91中的病毒点突变与HCC，干扰素（IFN）治疗失败和胰岛素抵抗的发展有关。这些病毒突变为与HCV相关的肝细胞癌发生的机械研究提供了令人信服的起点。我们最近发现，这些突变大大改变了HCV基因的表达，并控制了两种先前未知的HCV蛋白，分别为70和91 minicore蛋白的水平（Eng等，2009，J Virol）。提出的实验检查了这些突变的影响，以及我们确定的其他两个与HCC相关的HCV突变（Fishman等，2009，Clin Cancer res）。目标I的目的是确定核心基因突变是否是在肝癌发展之前出现的，因此有可能促进肝细胞癌。将进行一项嵌套的病例对照研究，以比较开发HCC的核心基因序列与在丙型肝炎抗病毒长期治疗抗肝硬化（HALT-C）试验期间未发展HCC的“病例”的核心基因序列。主要结果将是病例与对照组的百分比，其中病毒RNA种群中的大多数变体（即准特性）具有密码子70突变。将进行多变量分析，以确定密码子70突变是否是入射HCC的独立预测指标。次要结果将包括其他密码子和临床结果之间的突变之间的关联，以及准时间随着时间的推移的变化。 AIM II的实验目标是确定与HCC相关的突变在暴露于细胞培养的IFN期间是否赋予HCV的生存优势。病毒RNA和蛋白质动力学将通过我们既定的方法随时间而通过QRT/PCR和Western blotting测量。这些研究将确定四个与HCC相关的突变对HCV RNA复制，蛋白质合成（包括Minicore合成）以及HCV承受IFN治疗的能力的影响。米孔的体内表达将通过分析肝样本来确认。该项目挑战了美国研究人员广泛控制的范式，即所有HCV变体都具有同等的致癌潜力。我们的目标的实现将填补与HCV相关肿瘤发生的分子基础的基本空白，并可能导致鉴定用于治疗干预措施的新靶标，并为具有致癌变异的患者开发诊断测试。 公共卫生相关性：丙型肝炎病毒（HCV）是肝细胞癌（HCC）的主要原因，这是世界上与癌症相关死亡率的第三大最常见原因。临床研究表明，HCV核心基因中的点突变增加了HCC的风险并增加了干扰素治疗失败的可能性。由于它们的临床相关性，我们将定义四个与HCC相关突变对病毒复制的分子作用，并确定这些突变是否在HCC之前引起，因此保证研究可能使用毒性菌株的生物标志物，这些毒物可用于鉴定患有肝癌升高的患者。