Developing Murine Models of HCV Replication

描述（由申请人提供）：由于约2%的世界人口感染，丙型肝炎病毒（HCV）已成为一个重大的公共卫生问题，造成重大的医疗，社会和经济负担。 70- 90%的感染者未能清除病毒，并保持慢性感染，可能进展为持续性肝炎，肝硬化和肝细胞癌（HCC）。 因此，HCV相关肝病是美国肝移植的主要原因。 目前还没有预防性疫苗，只有20-30%的慢性感染患者对目前可用的治疗有反应。 因此，迫切需要预防性疫苗和替代治疗方案。 不幸的是，研究工作，以了解，从而打击，这种感染已受到阻碍，缺乏强大的组织培养复制系统和小动物模型。 虽然最近开发的体外HCV复制子系统克服了其中的一些限制，但仍需要小动物模型来研究病毒-宿主相互作用，以评估病毒的病理作用并更好地理解HCV免疫学（例如，恢复与持续的免疫学和病毒学基础），以及用于测试生理学和药理学试剂控制HCV感染的体内潜力。 因此，本探索性提案的目的是创建能够表达和潜在复制HCV的小鼠模型。 最初的努力将集中在使用可选择的HCV复制子，这已被证明是一个有效的HCV复制系统。 然而，在这些研究中获得的小鼠适应信息可能随后允许开发复制天然HCV基因组的类似模型。 具体目的是：1）确定是否可以通过调整目前可用的HCV复制子以在小鼠肝细胞中复制来改变HCV物种嗜性; 2）确定是否可以通过HCV构建体的瞬时T7驱动表达在体内建立急性HCV的鼠模型;和3）确定HCV构建体的稳定RNA聚合酶I驱动的表达是否可以用作体内HCV的慢性鼠模型。

DESCRIPTION (provided by applicant): With approximately 2% of the world population infected, Hepatitis C Virus (HCV) has emerged as a significant public health problem creating a significant medical, social, and economic burden. Between 70- 90% of those infected fail to clear the virus and remain chronically infected with the likelihood of progression to persistent hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). As a result, HCV-associated liver disease is the leading cause of liver transplantation in the United States. At this time, no preventative vaccine is available, and only 20-30% of chronically infected patients respond to the currently available therapy. Hence, there is a pressing need for a preventative vaccine and alternative treatment options. Unfortunately, research efforts to understand, and thus combat, this infection have been hindered by the lack of robust tissue culture replication systems and small animal models. While the recent development of the in vitro HCV replicon system has overcome some of these limitations, small animal models are still needed for the study of viral-host interactions to assess the pathological effects of the virus and to better understand HCV immunology (e.g. the immunological and virological basis of recovery versus persistence), as well as for testing the in vivo potential of physiological and pharmacological agents for controlling HCV infection. Therefore, the objective of this exploratory proposal is to create mouse models capable of expressing, and potentially replicating, HCV. Initial efforts will focus on the use of selectable HCV replicons, which have proven to be an efficient HCV replication system. However, the mouse-adaptation information gained in these studies may subsequently allow for the development of analogous models that replicate native HCV genomes. The Specific Aims to be addressed are: 1) determine if HCV species tropism can be altered by adapting currently available HCV replicons to replicate in mouse hepatocytes; 2) determine if a murine model of acute HCV can be established by transient T7-driven expression of HCV construct(s) in vivo; and 3) determine if stable RNA Polymerase I-driven expression of HCV construct(s) can serve as a chronic murine model of HCV in vivo.