Functional Analysis of the Hepatitis C Virus Genome

描述(申请人提供)：在过去一年中，我的实验室 建立丙型肝炎病毒亚基因组合成的组织培养体系 最初由Lohmann及其同事描述的复制体(Science 286：113， 1999年)。在这个系统的帮助下，我们已经获得了对 宿主-病毒相互作用与控制丙型肝炎病毒的抗病毒机制 复制。我们的研究计划现在的范围是利用这个系统 用于丙型肝炎病毒基因产物的功能分析和宿主的鉴定 影响病毒复制的决定因素，包括那些 由I型干扰素(干扰素)反应激活。我们的观察表明， 细胞周期可能在RNA病毒的复制中发挥关键作用， 我们的知识是史无前例的，将提供对主机病毒的新见解 互动。此外，我们将开发程序来酶法分离 来自感染细胞的活性复制复合体 识别在病毒复制中起作用的宿主因素。结果 不仅有助于更好地了解 病毒RNA合成，但也产生了关于宿主决定因素的新信息 控制丙型肝炎病毒的组织嗜性和宿主范围。其他实验包括 建议用于研究控制观察到的 丙型肝炎病毒分离株对干扰素-α的耐药性。为此，我们寻求开发 允许分离出具有IFN抗性的病毒变异株的条件 然后用于鉴定控制干扰素的病毒基因产物 抵抗。除了我们的整体方法外。我们还将建立一个详细的 病毒非结构区的遗传和功能图谱。可得性 这将为成功地进行 此应用程序中建议的实验。取得了预期的效果 从这个研究项目中，我们不仅将获得关于 丙型肝炎病毒复制的机制，但将进一步促进 确定抗病毒治疗的新靶点。丙型肝炎病毒是一种 可能导致急性和慢性肝炎并导致 肝细胞癌。原发丙型肝炎病毒感染尚不能通过 接种疫苗，到目前为止，有效和负担得起的治疗方法，以治愈 目前还没有超过1.7亿名携带者感染的情况。

DESCRIPTION (provided by applicant): During the past year my laboratory established a tissue culture system for the synthesis of subgenomic HCV replicons as originally described by Lohmann and colleagues (Science 286:113, 1999). With the help of this system, we have already gained new insights into host-virus interactions and the antiviral mechanisms that control HCV replication. The scope of our research program is now to exploit this system for a functional analysis of HCV gene products and identification of the host determinants that influence viral replication, including those that are activated by the type I interferon (IFN) response. Our observations that the cell cycle could play a critical role in replication of an RNA virus are, to our knowledge, unprecedented and will provide new insight into host-virus interactions. Furthermore, we will develop procedures to isolate enzymatically active replication complexes from infected cells for the subsequent identification of host factors playing a role in viral replication. Results from these investigations will not only contribute to a better understanding of viral RNA synthesis but also yield new information about host determinants that control tissue tropism and host-range of HCV. Additional experiments are proposed for the investigation of the mechanisms that control the observed resistance of HCV isolates to IFN-a. For this purpose we seek to develop conditions permitting the isolation of IFNresistant viral variants which can then be used for the identification of viral gene products controlling IFN resistance. Apart from our overall approach. we will also establish a detailed genetic and functional map of the viral non-structural region. the availability of which will provide a valuable tool for the successful conduct of the experiments proposed in this application. With the anticipated results obtained from this research program we will not only gain significant information about the mechanism of HCV replication, but will further contribute to the identification of novel targets for antiviral therapy. HCV is a pathogen of global significance that can cause acute and chronic hepatitis and induce hepatocellular carcinoma. Primary HCV infection cannot yet he prevented by vaccination and, so far, effective and affordable treatments to cure the infection in over 170 million carriers is not yet available.