Transgenic mouse models of hepatitis C virus replication

描述（由申请人提供）：丙型肝炎病毒（HCV）是慢性肝炎的常见原因，在美国感染了大约400万人，全世界感染了1.7亿人。绝大多数hcv感染者发展为慢性丙型肝炎，可发展为纤维化、肝硬化和肝细胞癌。由于缺乏HCV复制和感染的小动物模型，对HCV复制和肝癌发生的分子机制的研究一直受到阻碍。目前，黑猩猩是唯一可靠的易受生产性HCV感染和复制的动物。然而，黑猩猩模型的应用受到资源有限和维护成本高的制约。我们开发替代性HCV复制小动物模型的能力受到HCV感染宿主范围狭窄的限制。最近，我们在HCV复制的遗传研究和HCV复制和感染的小鼠模型的开发方面取得了重大进展。我们已经证明，来自染色体整合的HCV cDNA的HCV RNA导致感染性HCV的强劲生产。更重要的是，我们发现HCV RNA在小鼠胚胎成纤维细胞（mef）中有效地复制。这些进展使我们能够假设dna衍生的HCV RNA可以在转基因小鼠中有效地复制。本应用程序的总体目标是开发用于HCV RNA复制的转基因小鼠模型，并证明HCV RNA在转基因小鼠中的复制。因此，我们将开发多种转基因小鼠系，其中包含亚基因组和全长HCV rna的cdna以及可逆四环素控制的转录激活因子（rtTA）和抑制因子（tTS）。HCV RNA的转录表达受四环素应答启动子的控制。HCV RNA在转基因小鼠中的复制将通过多学科方法确定，并将通过使用HCV蛋白酶抑制剂和小鼠干扰素进行验证。发展概括HCV RNA复制、疾病发生和进展以及肝癌发生的动物模型将有助于确定宿主和病毒因子在HCV复制、发病机制和癌变中的作用和分子机制。此外，转基因HCV模型的可用性将加速针对HCV感染的预防性和治疗性抗病毒药物和疫苗的发现和开发，从而预防HCC的发展。

DESCRIPTION (provided by applicant): Hepatitis C virus (HCV) is a common cause of chronic hepatitis, infecting approximately 4 million people in the United States and 170 million people worldwide. The vast majority of HCV-infected individuals develop chronic hepatitis C, which can progress to fibrosis, cirrhosis, and hepatocellular carcinoma. The study of the molecular mechanisms of HCV replication and hepatocarcinogenesis has been hampered by the lack of a small animal model of HCV replication and infection. Currently, chimpanzee is the only reliable animal susceptible to productive HCV infection and replication. However, the application of chimpanzee model is restricted by its limited resource and high maintenance cost. Our ability to develop alternative small animal models for HCV replication is constrained by the narrow host range of HCV infection. Recently, we have made significant advances in genetic studies of HCV replication and development of small murine models of HCV replication and infection. We have demonstrated that HCV RNA derived from a chromosomally integrated HCV cDNA resulted in robust production of infectious HCV. More importantly, we have found that HCV RNA replicated efficiently in the mouse embryo fibroblasts (MEFs). These advances allow us to hypothesize that the cDNA-derived HCV RNA can replicate efficiently in transgenic mice. The overall goal of this application is to develop transgenic mouse models for HCV RNA replication and to demonstrate HCV RNA replication in transgenic mice. Thus, we will develop multiple transgenic mouse lines that contain the cDNAs of a subgenomic and full-length HCV RNAs and a reversible tetracycline-controlled transcriptional activator (rtTA) and suppressor (tTS). Transcriptional expression of HCV RNA is under the control of a tetracycline-responsive promoter. HCV RNA replication in transgenic mice will be determined by multidiscipline approaches and will be validated by the use of HCV protease inhibitors and mouse interferons. Development of animal models that recapitulate HCV RNA replication, disease development and progression, and hepatocarcinogenesis will facilitate the determination of the roles and molecular mechanisms of host and viral factors in HCV replication, pathogenesis, and carcinogenesis. Moreover, the availability of transgenic HCV models will accelerate the discovery and development of prophylactic and therapeutic antiviral drugs and vaccines against HCV infection and thereby prevent HCC development.