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: 10328917
• 负责人: Amit Kapoor
• 金额: $ 47.85万
• 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 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10328917
• 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; end stage liver disease; 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. !

项目摘要(摘要) 约有1.7亿人慢性感染丙型肝炎病毒(HCV)。病毒导致 每年大约有100万人死亡，200-300万人新感染。丙型肝炎病毒确立慢性嗜肝感染 在大多数受感染的个体中。然而，并不是所有慢性感染者都会患上肝病。这个 病毒和宿主因素及其相互作用，决定了丙型肝炎病毒的持久性和肝脏的发展 疾病仍然不清楚，主要是因为缺乏适当的小动物模型。值得注意的是，丙型肝炎病毒诱导 在人类体内持续数年或数十年的病毒感染后，肝脏疾病就会发生。 因此，一个信息性的肝病模型应该发展出长期的病毒持久性，从而导致慢性 肝脏炎症和疾病。只有黑猩猩才能作为研究丙型肝炎病毒感染的免疫活性宿主 但都不可用。考虑到丙型肝炎病毒严格的物种取向，最好的选择是开发一个替代品 病毒及其在宿主中的感染反映了人类和黑猩猩感染丙型肝炎病毒的模型。GBV-B 作为一种信息丰富的代孕模型，但它主要导致急性解体感染(几个月)。啮齿动物 肝炎病毒(HV)是丙型肝炎病毒的动物同源物。我们从野鼠(Rattus Norveicus)身上分离到一株RHV， 编号为RHV-RN1(初步数据)。我们确定RHV-rn1具有丙型肝炎病毒的定义特征： 嗜肝性，倾向终生坚持，进展缓慢的肝病。引起的免疫反应 RHV-rn1与丙型肝炎病毒相似，包括在急性期肝组织中诱导干扰素刺激的基因。 与慢性病毒感染相结合，我们表明免疫活性大鼠的RHV-RN1感染是一种 研究丙型肝炎病毒相关持久性和致病机制的创新模型。我们建议将宿主描述为 决定病毒感染结果和疾病的因素。因为，他们更好的特征对于 开发预防病毒持久性和疾病的新方法。目标1是研究病毒的自然历史 导致不同感染结局和肝脏的先天免疫反应的发病机制和动力学 病理学。AIM-2是感染大鼠肝脏内天然免疫反应的高分辨率图谱。我们会 用细胞类型特异性和单细胞RNA-SEQ分析研究感染病毒的系列基因表达变化 未感染的肝细胞、肝窦内皮细胞、枯否细胞和自然杀伤细胞。目标3是瞄准目标 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/去抑制病毒 抑制宿主基因表达。因此，我们将了解目标主机响应在 肝炎病毒的持久性和发病机制。我们相信，拟议的研究将提供前所未有的 深入了解在丙型肝炎病毒致病过程中宿主反应的体内相关性。此外，还可以进一步 我们新的完全免疫活性和易处理的模型的特征将允许开发创新的 预防病毒持久性和肝病的手段。 好了！