Functional Analysis of the Hepatitis C Virus Genome

描述（由申请人提供）：在过去的一年里，我的实验室 建立了合成亚基因组HCV的组织培养体系 复制子最初由Lohmann和同事描述（Science 286：113， 1999年）。在这个系统的帮助下，我们已经获得了新的见解， 宿主-病毒相互作用和控制HCV的抗病毒机制 复制的我们的研究计划的范围现在是利用这个系统 用于HCV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（IFN）应答激活。我们观察到， 细胞周期可能在RNA病毒复制中起关键作用， 我们的知识，前所未有的，并将提供新的洞察主机病毒 交互.此外，我们将开发酶法分离的程序， 感染细胞的活性复制复合物， 鉴定在病毒复制中起作用的宿主因子。结果 这些研究不仅有助于更好地了解 病毒RNA合成，而且还产生了关于宿主决定簇的新信息， 控制HCV的组织嗜性和宿主范围。其他实验包括 提出的调查机制，控制观察到的 HCV分离株对IFN-α的抗性。为此，我们致力于发展 允许分离IFN抗性病毒变体的条件， 然后用于鉴定控制IFN的病毒基因产物 阻力除了我们的整体思路。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。的可用性 这将为成功地举行联合国 本申请中提出的实验。取得了预期的成果 从这项研究计划中，我们不仅将获得有关 HCV复制的机制，但将进一步有助于 抗病毒治疗新靶点的鉴定。HCV是一种病原体， 全球意义，可引起急性和慢性肝炎，并诱导 肝细胞癌原发性HCV感染尚不能通过 疫苗接种，以及迄今为止有效和负担得起的治疗方法， 超过1.7亿携带者的感染情况尚未掌握。

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.