Genetic Viral and Host Adaptations to Breach Species Barriers of HCV

突破 HCV 物种屏障的遗传病毒和宿主适应

• 批准号: 10202407
• 负责人: Alexander Ploss
• 金额: $ 47.48万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10202407
• 关键词: Aftercare; Alcohol consumption; Animal Model; Antiviral Agents; Cells; Centers for Disease Control and Prevention (U.S.); Cercopithecidae; Cessation of life; Chronic Hepatitis C; Cirrhosis; Communicable Diseases; Complement; Cyclophilin A; Data; Development; Diabetes Mellitus; Distant; Drug usage; Fibrosis; Germany; HCV Animal Models; HIV; Health Services Accessibility; Hepatitis C; Hepatitis C Therapy; Hepatitis C Transmission; Hepatitis C virus; Hepatitis C-Like Viruses; Hepatocyte; Human; Imaging Techniques; Immune response; In Vitro; Individual; Infection; Integration Host Factors; Knock-in Mouse; Lead; Liver Stem Cell; Liver diseases; Malignant neoplasm of liver; Mus; Natural Immunity; Obesity; Orthologous Gene; Pan Genus; Patients; Peptidylprolyl Isomerase; Pongidae; Primary carcinoma of the liver cells; Primates; Public Health; Publishing; RNA; RNA replication; Reporting; Research; Rodent; Seminal; Shapes; Testing; Tissues; Transgenic Mice; Treatment Cost; Tropism; Vaccines; Viral; Viral Pathogenesis; Virus; Virus Replication; Work; advanced disease; cancer risk; chronic infection; co-infection; comorbidity; high risk; improved; in vivo; insight; liver transplantation; loss of function; novel; pathogen; prevent; receptor; response; single-cell RNA sequencing; stem cell biology; stem cells; transcriptomics; transmission process; vaccine candidate; vaccine evaluation; virus genetics

Project summary Hepatitis C virus (HCV) is an important and underreported infectious disease, causing chronic infection in ~71 million people worldwide. The CDC estimates there are ~30,000 new cases of HCV every year in the US and about 20,000 deaths annually, making HCV more deadly than 60 other infectious diseases combined, including HIV. However, the underlying mechanisms that lead to chronic HCV infection followed by end-stage liver disease are poorly understood. Although chronic hepatitis C can now be effectively treated with direct-acting antivirals (DAAs), a vaccine to prevent transmission remains a high priority due to extremely high treatment costs ($80,000+ for a 12-week course). Furthermore, once infected, individuals remain at high risk for liver disease even post-treatment. The proposed work will build on our substantial research findings to continue systematically analyzing the mechanisms that create barriers to interspecies HCV transmission. Aim 1: Define mechanisms of known host factors from diverse species that support HCV replication. HCV relies on a variety of host factors to establish replication in hepatocytes. Our preliminary data demonstrate that for all great apes tested, orthologs of peptidylprolyl isomerase A, also known as cyclophilin A (CypA), support HCV RNA replication. However, CypA of distantly related species, such as New and Old World monkeys and mice, is far less efficient. We aim to define mechanistically the underlying incompatibility of this and other host factors with the virally encoded components of the HCV replication machinery. Aim 2: Characterize HCV infection and immune response networks following infection across primate and rodent species. We will study whether HCV can infect and replicate in a novel set of stem cell-derived hepatocyte-like cells from a range of evolutionarily diverse species, including great apes, selected New and Old World monkeys and rodents. In a second step, we will use high-throughput single-cell RNA sequencing to derive species-specific transcriptomic response networks associated with HCV replication. Aim 3: Characterize the impact of CD302 and Cr1L on restricting HCV infection in vivo. Human entry factor transgenic mice with blunted innate immunity only support low-level viral replication, suggesting murine restriction factors may suppress viral replication in vivo. Our preliminary data indicate mouse CD302 and complement component (3b/4b) receptor 1-like (Cr1L) limit HCV replication in vitro. Here, we will assess if loss- of-function of mCD302 and mCr1L augments HCV infection in our HCV entry factor knock-in mice. The proposed work will provide new insights into the species tropism of HCV. The Ploss lab has made many seminal contributions to the field and will be aided in this important work by our long-standing collaborators Drs. Schwartz (Weill Cornell), Shalek (MIT) and Pietschmann (Twincore, Germany). Our work will advance the field of HCV research by making progress in the development of small animal models suitable for studying HCV infection and immune responses, a necessary precursor to improving treatment and developing vaccines.

项目概要 丙型肝炎病毒 (HCV) 是一种重要且报告不足的传染病，导致约 71 人慢性感染 全球数百万人。 CDC 估计美国每年约有 30,000 例新发 HCV 病例 每年约 20,000 人死亡，使丙型肝炎病毒比 60 种其他传染病的致死率加起来还要高，其中包括 艾滋病病毒。然而，导致慢性 HCV 感染继而导致终末期肝病的潜在机制 人们了解甚少。尽管慢性丙型肝炎现在可以用直接作用的抗病毒药物有效治疗 （DAAs），由于治疗成本极高，预防传播的疫苗仍然是重中之重 （为期 12 周的课程费用超过 80,000 美元）。此外，一旦感染，个体仍处于患肝病的高风险中 甚至治疗后。拟议的工作将以我们大量的研究成果为基础，系统地继续进行 分析为种间 HCV 传播造成障碍的机制。 目标 1：定义来自不同物种的支持 HCV 复制的已知宿主因子的机制。 HCV 依赖多种宿主因子在肝细胞中建立复制。我们的初步数据表明 对于所有测试的类人猿，肽基脯氨酰异构酶 A（也称为亲环蛋白 A (CypA)）的直向同源物支持 HCV RNA 复制。然而，远亲物种的 CypA，例如新世界猴和旧世界猴， 老鼠，效率就低得多。我们的目标是机械地定义该主机和其他主机的潜在不兼容性 HCV 复制机制的病毒编码成分的因子。 目标 2：表征灵长类动物 HCV 感染和感染后的免疫反应网络 和啮齿动物种类。我们将研究丙型肝炎病毒是否可以在一组新的干细胞衍生的细胞中感染和复制。 来自一系列进化多样的物种的肝细胞样细胞，包括类人猿，精选的新旧物种 世界猴子和啮齿动物。第二步，我们将使用高通量单细胞 RNA 测序来衍生 与 HCV 复制相关的物种特异性转录组反应网络。 目标 3：表征 CD302 和 Cr1L 对限制体内 HCV 感染的影响。人为进入因素 先天免疫减弱的转基因小鼠仅支持低水平的病毒复制，这表明小鼠 限制因子可能会抑制病毒在体内的复制。我们的初步数据表明小鼠 CD302 和 补体成分 (3b/4b) 受体 1 样 (Cr1L) 限制 HCV 体外复制。在这里，我们将评估损失是否- mCD302 和 mCr1L 的功能丧失会增强 HCV 进入因子敲入小鼠的 HCV 感染。 拟议的工作将为 HCV 的物种向性提供新的见解。 Ploss 实验室做了很多 对该领域的开创性贡献，并将在这项重要工作中得到我们的长期合作者博士的协助。 Schwartz（威尔·康奈尔）、Shalek（麻省理工学院）和 Pietschmann（Twincore，德国）。我们的工作将推动该领域的发展 通过在适合研究 HCV 的小动物模型的开发方面取得进展，促进 HCV 研究 感染和免疫反应，这是改善治疗和开发疫苗的必要前提。