Modulation of HCV Gene Expression

描述(申请人提供)：丙型肝炎病毒(丙型肝炎病毒)对公众健康构成威胁。重要的是要找出阻碍自然防御并导致治疗失败的病毒因素。在一项横断面研究中，我们最近发现病毒复制和针对丙型肝炎病毒新的交替阅读框架蛋白(ARFP)的抗体的存在之间存在反向关系。ARFP是本课题组以前发现的。抗ARFP抗体的峰值表明，当丙型肝炎病毒复制被抑制时，ARFP的产生增加。抗ARFP抗体阳性患者的丙型肝炎病毒RNA在一个被称为末端干环的顺式调节元件(CRE)中发生突变，这是我们之前也发现的一种RNA结构。我们最近的结果表明，ARFP可能有助于丙型肝炎病毒在不利条件下生存，并表明Cres可能控制ARFP的产生。拟议的研究是基于我们最近的数据，以及其他人的数据，表明晚期肝硬变和肝癌患者的丙型肝炎病毒RNA在末端干环发生突变。 Aim I的实验验证了这样一种假设，即干扰素/利巴韦林治疗诱导的有效抗病毒反应的显著特征是对ARFP的免疫反应增加。比较治疗前、治疗中、治疗后丙型肝炎病毒RNA和抗ARFP抗体水平、外周血单个核细胞刺激指数(SI)和表型。 目标2中的实验验证了这一假设，即在干扰素/利巴韦林治疗期间以及在进展为晚期肝病和肝癌的过程中，末端干环元件的突变会累积，调节其功能并增加ARFP的产生。将开发新的软件并将其应用于扩大的突变数据集，并将确定末端茎环的结构/功能关系。 这些研究将提供有关丙型肝炎病毒适应机制和基因表达途径的新信息，并将推动丙型肝炎病毒新药和疫苗的开发。

DESCRIPTION (provided by applicant): The hepatitis C virus (HCV) poses a public health threat. It is important to identify viral factors that thwart natural defenses and contribute to treatment failure. In a cross sectional study, we recently found an inverse relationship between viral replication and the presence of antibodies directed against HCV's novel alternate reading frame proteins (ARFPs). ARFPs were discovered previously by our group. The spike of anti-ARFP antibodies suggests that production of ARFPs is increased when HCV replication is inhibited. HCV RNAs of patients with anti-ARFP antibodies have mutations in a putative cis regulatory element (CRE) called the Terminal Stem-loops, an RNA structure we also discovered previously. Our recent results suggest that ARFPs may help HCV survive adverse conditions, and indicate that CREs may control the production of ARFPs. The proposed studies are based on our recent data, and on data of others showing that HCV RNAs of patients with advanced cirrhosis and liver cancer have mutations in the Terminal Stem-loops. Experiments in Aim I test the hypothesis that effective anti-viral responses induced by IFN/ribavirin treatment are marked by a rise in immune responses to ARFPs. HCV RNA and anti-ARFP antibody levels, and the stimulation index (SI) and phenotype of peripheral blood mononuclear cells will be compared before, during, and after treatment. Experiments in Aim 2 test the hypothesis that mutations in the Terminal Stem-loops element accumulate during IFN/ribavirin treatment and during progression to advanced liver disease and liver cancer, modulating its function and increasing production of ARFPs. New software will be developed and applied to an expanded dataset of mutations, and the structure/function relationships of the Terminal Stem-loops will be defined. New information about HCV's adaptive mechanisms and pathways of gene expression will emerge from these studies and will advance the development of new drugs and vaccines for HCV.