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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

基本信息

批准号：
9104080
负责人：
Alexander Ploss
金额：
$ 24.86万
依托单位：
PRINCETON UNIVERSITY
依托单位国家：
美国
项目类别：
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/9104080
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 Health Hepatitis B Hepatitis B Virus Hepatitis C Hepatitis C virus Hepatitis Viruses Hepatocyte Human Immune response 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 Publishing RNA Viruses RNA replication 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 antiviral immunity clinically relevant gene product human morbidity human mortality improved induced pluripotent stem cell insight interest laser capture microdissection nonhuman primate novel occludin pathogen permissiveness repository response species difference success transcriptomics uptake viral RNA virology

项目摘要

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（由申请人提供）：传染病对人类发病率和死亡率有重大影响。许多引起人类疾病的病原体的宿主范围非常狭窄，这给研究和开发有效的临床疗法带来了挑战。我们将重点研究丙型肝炎病毒（HCV），这是一种宿主范围狭窄的典型嗜肝病毒，并得出影响跨物种感染结果的细胞内在免疫反应模式。百分之八十的丙型肝炎病毒感染会导致慢性感染，并可能导致肝硬化和肝细胞癌。 HCV 具有不完全确定的宿主向性，几乎仅局限于人类和黑猩猩。我们之前已经证明，两种宿主因子 CD81 和 occludin (OCLN) 的氨基酸序列之间的差异（人和小鼠之间）会阻止小鼠对 HCV 的摄取。虽然从人类到啮齿动物等多种物种的肝细胞都支持 HCV 组装和释放，但抗病毒先天反应似乎限制了非人类物种中 HCV RNA 的复制。我们假设抗病毒基因产物的动力学、多样性和大小方面的物种特异性差异会影响病毒的趋向性。在这里，我们建议开发和利用一个新的平台来系统地定义这种有助于感染结果的物种特异性反应模式。我们的目标是利用非人类灵长类动物的真皮成纤维细胞来产生诱导多能干细胞 (iPSC)，并逐步将这些猿 iPSC 分化为肝细胞样细胞 (HLC)。我们将描述和比较来自人类、黑猩猩、倭黑猩猩、大猩猩、猩猩和恒河猴标本的 HLC 中的病毒感染。对于支持病毒生命周期至关重要的重要宿主因子的直向同源物在这些物种中很大程度上是保守的，易感性的差异可能归因于抗病毒反应模式的差异和/或HCV无法拮抗细胞内在防御。我们将 HCV 感染依赖性荧光重定位 (HDFR) 报告基因与激光捕获显微切割 (LCM) 结合起来，从不同物种的 HCV 感染 HLC 中分离出感染细胞，并对它们进行转录组分析。通过比较和对比感染频率高和低的物种的数据，我们将能够确定与有效抗病毒控制相关的物种特异性特征。该合作项目的成功取决于 HCV 病毒学和病毒向性（Ploss 实验室）以及干细胞和肝细胞生物学（Schwartz 实验室）方面的互补专业知识。我们相信，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，并揭示抗病毒免疫对限制物种向性的影响。由于我们收集材料的许多灵长类物种都濒临灭绝，并且不能（不再）用于生物医学研究，因此我们将建立的新兴 iPSC 存储库也将为其他需要访问通常无法访问的灵长类物种/组织的研究人员提供支持平台。