Host and viral determinants of hepatitis C virus macaque infection

丙型肝炎病毒猕猴感染的宿主和病毒决定因素

• 批准号: 9012008
• 负责人: Matthew J Evans
• 金额: $ 26.36万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-10 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9012008
• 关键词: Animal Model; Animal Testing Alternatives; Animals; CD81 gene; Cell Culture Techniques; Cells; Chimera organism; Defect; Detection; Development; Disease; Enzymes; Ethics; Exhibits; Figs - dietary; Genetic; Genotype; Goals; Grant; HCV Animal Models; HIV-1; Health; Hepatic; Hepatitis C; Hepatitis C Antibodies; Hepatitis C Vaccine; Hepatitis C virus; Hepatocyte; Human; Immunity; Immunocompetent; Infection; Inflammation; Lead; Left; Liver; Liver diseases; Macaca; Macaca mulatta; Macaca nemestrina; Malignant neoplasm of liver; Modeling; Monitor; Monkeys; Mus; Mutation; North America; Pan Genus; Pathogenesis; Patients; Play; Primary carcinoma of the liver cells; Primates; Process; Proteins; Reagent; Research; Research Personnel; Risk; Role; Sampling; Serum; Structural Genes; Supporting Cell; Testing; Time; Tropism; United States; United States National Institutes of Health; Vaccine Design; Vaccines; Variant; Viral; Virus; Virus Receptors; Virus Replication; Work; arm; base; clinically relevant; combat; fitness; in vivo; induced pluripotent stem cell; insight; liver injury; mutant; pathogen; research study; screening; tool; vaccine evaluation; viral RNA; virus genetics; virus pathogenesis

 DESCRIPTION (provided by applicant): The hepatitis C virus (HCV) is the leading cause of liver cancer in the Western Hemisphere. Over 170 million people are infected with this virus and consequently at increased risk of liver damage and cancer. Understanding how this virus induces liver disease and the development of a much-needed vaccine requires an immunocompetent HCV animal model. The dogma in the HCV field was that this virus only infects humans and chimpanzees. As a recent NIH moratorium on chimpanzee research has effectively prohibited use of these animals, an alternative immunocompetent HCV animal model is desperately needed. To explore if small primates can support HCV infection, we generated hepatocytes from pigtail macaque-(Macaca nemestrina, Mn) induced pluripotent stem cell-derived hepatic cells. Surprisingly, these cells supported detectable levels of infection with cell culture derive genotype 2a HCV. We found that inefficient function of the macaque version of the HCV receptor, CD81, limited HCV infection of these cells. A mutant genotype 2a virus that can use CD81 proteins from a wide range of species overcame this block and efficiently infected the macaque hepatocytes. We have recently made the startling discovery that both wild type and mutant versions of this virus could infect pigtail macaques in vivo, thus demonstrating for the first time that HCV can replicate in animals other than humans and chimpanzees. In this grant we propose experiments to further characterize macaque HCV infections. This will include determining if rhesus macaques, which are more readily available and a larger set of species-specific reagents than pigtail macaques are obtainable, can also be infected with genotype 2a HCV. We will also examine the viral determinants for HCV infection of macaques. We will explore mechanisms by which virus can adapt to more efficiently replicate in this host and determine if we are able to expand the range of viral isolates that can infect NHPs. Ultimately, our studies will provide the first immunocompetent tractable animal models that will establish new avenues to explore HCV replication and pathogenesis in vivo with the eventual goal of developing efficacious vaccine against this virus.

 描述（由申请人提供）：丙型肝炎病毒（HCV）是西半球肝癌的主要原因。超过1.7亿人感染了这种病毒，因此肝损伤和癌症的风险增加。了解这种病毒如何诱导肝脏疾病和急需的疫苗的发展需要一个免疫活性的HCV动物模型。丙型肝炎病毒领域的教条是，这种病毒只感染人类和黑猩猩。由于NIH最近暂停了对黑猩猩的研究，有效地禁止了这些动物的使用，因此迫切需要一种替代的免疫活性HCV动物模型。为了探索小灵长类动物是否可以支持HCV感染，我们从猪尾猕猴（Macaca nemestrina，Mn）诱导的多能干细胞衍生的肝细胞产生肝细胞。令人惊讶的是，这些细胞支持可检测水平的细胞感染， 培养衍生基因型2a HCV。我们发现，效率低下的猕猴版本的HCV受体，CD81，限制HCV感染这些细胞。一种突变的基因型2a病毒，可以使用来自广泛物种的CD81蛋白，克服了这一障碍，并有效地感染了猕猴肝细胞。我们最近有了惊人的发现，这种病毒的野生型和突变型都可以在体内感染猪尾猕猴，从而首次证明HCV可以在人类和黑猩猩以外的动物中复制。在这项资助中，我们提出了进一步描述猕猴丙型肝炎病毒感染特征的实验。这将包括确定恒河猴是否也可以感染基因型2a HCV，恒河猴比猪尾猴更容易获得和更大的物种特异性试剂。我们还将研究猕猴HCV感染的病毒决定因素。我们将探索病毒在宿主中更有效复制的机制，并确定我们是否能够扩大感染NHP的病毒分离株的范围。最终，我们的研究将提供第一个具有免疫活性的易处理的动物模型，这将建立新的途径来探索HCV在体内的复制和发病机制，最终目标是开发针对这种病毒的有效疫苗。