Host response and liver disease in a hepatitis C-like virus rat model

丙型肝炎样病毒大鼠模型中的宿主反应和肝病

• 批准号: 10084805
• 负责人: Amit Kapoor
• 金额: $ 48.6万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10084805
• 关键词: Acute; Acute Hepatitis; Animal Model; Animals; Biological; Cadherins; Cells; Cessation of life; Characteristics; Chronic; Chronic Hepatitis C; Complex; Data; Defect; Dependovirus; Development; Disease; Disease model; Endothelial Cells; Epithelial; Event; GB virus B; Gene Expression; Genes; Genetic; Goals; HCV Liver Disease; Hepatitis C; Hepatitis C virus; Hepatitis C-Like Viruses; Hepatocyte; Histologic; Human; ISG15 gene; Immune; Immune response; Immunocompetent; In Situ Hybridization; Individual; Infection; Inflammation; Innate Immune Response; Integration Host Factors; Interferon Activation; Interferons; Internet; Knowledge; Kupffer Cells; Life; Liver; Liver diseases; Lymphoid; MicroRNAs; Modeling; Morphology; Mus; Natural History; Natural Immunity; Natural Killer Cells; Nature; Outcome; Pan Genus; Pathogenesis; Pathology; Patients; Persons; Phase; Population; Rattus; Rattus norvegicus; Research; Resolution; Rodent; Role; Saguinus; Sampling; Signal Transduction; System; T-Lymphocyte; Technology; Testing; Time; Tissue-Specific Gene Expression; Triad Acrylic Resin; Tropism; Viral; Viral Genome; Viral Pathogenesis; Viral hepatitis; Viremia; Virus; Virus Diseases; Virus Replication; acute infection; adeno-associated viral vector; bile duct; cell type; chronic infection; chronic liver inflammation; differential expression; disease phenotype; in vivo; innovation; insight; novel; prevent; public health relevance; response; reverse genetics; single-cell RNA sequencing; transcriptome; transcriptome sequencing

Project Summary (Abstract) Approximately 170 million people are chronically infected with hepatitis C virus (HCV). The virus causes about a million deaths and 2-3 million new infections each year. HCV establishes chronic hepatotropic infection in the majority of infected individuals. However, not all chronically infected persons develop liver diseases. The viral and host factors, and their interactions, that determines the development of HCV persistence and liver diseases remains obscure largely due to the lack of an appropriate small animal model. Notably, HCV induced liver diseases develops after years or decades of persistent viral infection in humans, the natural host of HCV, and therefore an informative liver disease model should develop long term viral persistence leading to chronic liver inflammation and diseases. Only chimpanzees can serve as immunocompetent hosts to study HCV infection but are unavailable. Considering the strict species tropism of HCV, the best alternative is to develop a surrogate model where the virus and its infection in the host mirror HCV infection in humans and chimpanzees. GBV-B served as an informative surrogate model but it mostly causes acute resolving infection (of a few months). Rodent hepaciviruses (RHV) are animal homologs of HCV. We isolated a RHV from wild rats (Rattus norvegicus), designated RHV-rn1 (preliminary data). We determined that RHV-rn1 possesses HCV’s defining hallmarks: hepatotropism, propensity to life-long persistence and slow progressing liver disease. Immune responses elicited by RHV-rn1 and HCV are similar including the induction of interferon stimulated genes in the liver during acute and chronic viral infection Taken together, we showed that RHV-rn1 infection of immunocompetent rats is an innovative model to study HCV-related persistence and pathogenesis. We propose to characterize the host factors that dictate viral infection outcomes and diseases. Since, their better characterization is critical for developing new means of preventing viral persistence and diseases. Aim-1 is to study natural history of virus pathogenesis and dynamics of innate immune responses culminating in different infection outcomes and liver pathologies. Aim-2 is high-resolution mapping of innate immune responses in the liver of infected rats. We will study serial gene expression changes using cell-type specific and single-cell RNA-seq analysis of virus infected and uninfected hepatocytes, sinusoidal endothelial cells, Kupffer cells and natural killer cells. Aim-3 is to target the viral genomes and specific innate immune responses for achieving virus clearance or averting liver diseases. We will use miRNA expressing Adeno-associated virus (AAV) vectors to inhibit virus replication or repress/de- repress host gene expression. Consequently, we will know the precise role of the targeted host responses in hepacivirus persistence and pathogenesis. We believe that the proposed studies will provide unprecedented insights into the in vivo relevance of host responses during HCV pathogenesis. Additionally, further characterization of our novel fully immunocompetent and tractable model will allow development of innovative means to prevent viral persistence and liver diseases. !

项目概要（摘要） 大约 1.7 亿人慢性感染丙型肝炎病毒 (HCV)。病毒导致 每年约有 100 万人死亡和 2-300 万人新感染。 HCV 引起慢性嗜肝感染 在大多数感染者中。然而，并非所有慢性感染者都会患上肝脏疾病。这 病毒和宿主因素及其相互作用，决定了 HCV 持久性和肝脏的发展 疾病仍然不清楚，很大程度上是由于缺乏合适的小动物模型。值得注意的是，HCV 诱发 人类是 HCV 的自然宿主，经过数年或数十年的持续病毒感染后，会出现肝脏疾病， 因此，信息丰富的肝病模型应该发展长期的病毒持续存在，导致慢性 肝脏炎症和疾病。只有黑猩猩才能作为免疫活性宿主来研究丙型肝炎病毒感染 但不可用。考虑到HCV严格的物种向性，最好的选择是开发替代品 该模型中的病毒及其在宿主中的感染反映了人类和黑猩猩中的 HCV 感染。 GBV-B 作为一种信息丰富的替代模型，但它主要导致急性消退性感染（几个月）。啮齿动物 肝炎病毒（RHV）是HCV的动物同源物。我们从野生大鼠（Rattusnorvegicus）中分离出 RHV， 指定为 RHV-rn1（初步数据）。我们确定 RHV-rn1 具有 HCV 的定义特征： 促肝性、终生持续的倾向和缓慢进展的肝病。引发免疫反应 RHV-rn1 和 HCV 的作用相似，包括急性期肝脏中干扰素刺激基因的诱导 和慢性病毒感染 综合起来，我们发现免疫功能正常的大鼠的 RHV-rn1 感染是一种 研究 HCV 相关持续性和发病机制的创新模型。我们建议表征主机 决定病毒感染结果和疾病的因素。因为，更好的表征对于 开发预防病毒持续存在和疾病的新方法。 Aim-1是研究病毒的自然史 先天免疫反应的发病机制和动态最终导致不同的感染结果和肝脏 病理学。 Aim-2 是受感染大鼠肝脏先天免疫反应的高分辨率图谱。我们将 使用病毒感染的细胞类型特异性和单细胞 RNA-seq 分析来研究一系列基因表达变化 以及未感染的肝细胞、肝窦内皮细胞、库普弗细胞和自然杀伤细胞。 Aim-3 是瞄准 病毒基因组和特定的先天免疫反应，以实现病毒清除或避免肝脏疾病。 我们将使用表达 miRNA 的腺相关病毒 (AAV) 载体来抑制病毒复制或抑制/抑制病毒复制。 抑制宿主基因表达。因此，我们将知道目标宿主反应在 肝炎病毒的持续存在和发病机制。我们相信所提出的研究将提供前所未有的 深入了解 HCV 发病过程中宿主反应的体内相关性。另外，进一步 我们新型的完全免疫活性且易于处理的模型的表征将允许开发创新的 意味着预防病毒持续存在和肝脏疾病。 ！