Impact of species-specific host responses on restriction of hepatotropic viruses

物种特异性宿主反应对嗜肝病毒限制的影响

• 批准号: 8871022
• 负责人: Alexander Ploss
• 金额: $ 22.27万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8871022
• 关键词: Affect; Amino Acid Sequence; Animal Model; Antiviral Agents; Antiviral Response; Biology; Biomedical Research; Blood; Borneo; CD81 gene; Cell Line; Cell physiology; Cells; Cercopithecus pygerythrus; Cessation of life; Chronic; Cirrhosis; Clinical; Collection; Communicable Diseases; Communities; Cytochrome P450; Data; Dermal; Development; Disease; Exhibits; Family member; Fibroblasts; Fibrosis; Flavivirus; Fluorescence; Frequencies; Generations; Gorilla gorilla; HCV Animal Models; Hepatitis B; Hepatitis B Virus; Hepatitis C; Hepatitis C virus; Hepatitis Viruses; Hepatocyte; Human; Immune response; Immunity; Infection; Integration Host Factors; Investigation; Japanese Population; Kinetics; Lead; Life Cycle Stages; Light; Liver; Liver diseases; Macaca mulatta; Malignant neoplasm of liver; Mus; Orthologous Gene; Outcome; Pan Genus; Pan paniscus; Pan troglodytes; Papio; Pathway interactions; Pattern; Persons; Plague; Pluripotent Stem Cells; Pongidae; Pongo; Pongo pygmaeus; Predisposition; Primary carcinoma of the liver cells; Primates; Proteins; Publishing; RNA Viruses; RNA replication; Relative (related person); Reporter; Research Personnel; Resistance; Rodent; Role; Serum Proteins; Shapes; Signal Transduction; Skin; Specimen; Stem cells; System; Tissues; Tropism; Viral; Virus; Virus Assembly; Virus Diseases; Virus Replication; Work; clinically relevant; human morbidity; human mortality; improved; induced pluripotent stem cell; insight; interest; laser capture microdissection; nonhuman primate; novel; occludin; pathogen; permissiveness; public health relevance; repository; response; species difference; success; transcriptomics; uptake; viral RNA; virology

 DESCRIPTION (provided by applicant): Infectious diseases contribute significantly to human morbidity and mortality. Many pathogens that cause disease in humans, exhibit a very narrow host range, which poses challenges both for study and for developing effective clinical therapies. We will focus our studies on hepatitis C virus (HCV), an archetypal hepatotropic virus with a narrow host range and derive cell-intrinsic immune response patterns that impact infection outcome across species. Eighty percent of HCV infections result in chronic infection and can lead to cirrhosis and hepatocellular carcinoma. HCV has an incompletely defined host tropism limited almost exclusively to humans and chimpanzees. We have previously demonstrated that differences (between human and mouse) between the amino acid sequences of two host factors, CD81 and occludin (OCLN) block HCV uptake in mice. While HCV assembly and release is supported in hepatocytes across a broad variety of species ranging from humans to rodents, antiviral innate responses appear to limit HCV RNA replication in non-human species. We hypothesize that species-specific differences in the kinetics, diversity and magnitude of antiviral gene products influence viral tropism. Here, we propose to develop and utilize a novel platform to systematically define such species-specific response patterns contributing to infection outcome. We aim to capitalize on a collection of dermal fibroblasts from non-human primate species to generate induced pluripotent stem cells (iPSCs) and differentiate these simian iPSCs in a stepwise manner to hepatocyte like cells (HLCs). We will characterize and compare viral infections in HLCs derived from human, chimpanzee, bonobo, gorilla, orangutan and rhesus monkey specimens. Orthologs of essential host factors critical for supporting the viral life-cycle are largely conserved across these species and differences in susceptibility can likely be attributed to differences in antiviral response patterns and/or HCV's inability to antagonize cell-intrinsic defenses. We will couple a HCV infection-dependent fluorescence relocalization (HDFR) reporter with laser capture microdissection (LCM) to isolate infected cells from HCV infected HLCs from different species and subject them to transcriptomic profiling. By comparing and contrasting data from species with high and low infection frequencies, we will be able to determine species-specific signatures associated with effective antiviral control. The success of this collaborative project depends on the complementary expertise in HCV virology and viral tropism (Ploss lab) and stem cell and hepatocyte biology (Schwartz lab). We are convinced that this systems virology approach will provide rich insights into evolutionary conservation of antiviral response patterns and shed light on the impact of antiviral immunity to restrict species tropism. Since many of the primate species for which we collect materials are endangered and cannot be used (anymore) in biomedical research, the emerging iPSC repository we will build will also be an enabling platform for other researchers needing access to usually inaccessible primate species/tissues.

 描述（由适用提供）：传染病对人类的发病率和死亡率产生了重大贡献。许多引起人类疾病的病原体表现出非常狭窄的宿主范围，这对研究和开发有效的临床疗法构成了挑战。我们将把研究重点放在丙型肝炎病毒（HCV）上，这是一种型原型肝脏病毒，具有狭窄的宿主范围，并导致了影响整个物种感染结果的细胞内部免疫增强模式。 HCV感染的80％导致慢性感染，并可能导致肝硬化和肝细胞癌。 HCV几乎完全限制了人类和黑猩猩的不完全定义的宿主向手。我们先前已经证明，小鼠CD81和occludin（OCLN）的两个宿主因子的氨基酸序列之间的差异（人与小鼠之间）的差异。尽管HCV组装和释放在从人类到啮齿动物等各种物种的肝细胞中得到支持，但抗病毒先天反应似乎限制了非人类物种中的HCV RNA复制。我们假设在动力学，抗病毒基因产物的多样性和幅度上的物种特异性差异会影响病毒性偏向主义。在这里，我们建议开发和利用一个新型的平台，以系统地定义有助于感染结果的这种物种特定的反应模式。我们的目标是利用非人类灵长类动物的皮肤成纤维细胞的集合来产生诱导的多能干细胞（IPSC），并以逐步的方式将这些拟南芥IPSC分化为肝细胞类似细胞（HLC）。我们将表征和比较来自人类，黑猩猩，Bonobo，Gorilla，Gorilla，Orangutan和Rhesus Monkey标本的HLC中的病毒感染。基本宿主因素的直系因素在这些物种中很大程度上是在很大程度上保守的，并且易感性的差异可能归因于抗病毒反应模式的差异和/或HCV无法拮抗细胞中性防御。我们将将HCV感染依赖性荧光重新定位（HDFR）报告基因与激光捕获显微解剖（LCM）相结合，以将感染的细胞与来自不同物种的HCV感染的HLC隔离，并使它们接受转录组分析。通过比较和对比具有高感染频率和低感染频率的物种的数据，我们将能够确定与有效抗病毒药控制相关的物种特异性特异性特异性特征。该协作项目的成功取决于HCV病毒学和病毒疗法（Ploss Lab）和干细胞和肝细胞生物学（Schwartz Lab）的完整专家。我们坚信，这种系统病毒学方法将为抗病毒反应模式的进化保护提供丰富的见解，并阐明了抗病毒免疫学对限制物种tropism的影响。由于我们收集材料的许多灵长类动物是濒临灭绝的，并且在生物医学研究中不再使用（再也无法使用），因此我们将建立的新兴IPSC存储库也将是一个有能力的平台，对于需要访问通常无法接近的灵长类动物/组织的其他研究人员。