Cellular Programming in Persistent Versus Lytic Viral Infections

持续性与溶解性病毒感染中的细胞重编程 项目摘要 这项研究的长期目标是确定介导JC多瘤病毒的细胞因子 （JCPyV）感染，以更好地了解病毒-宿主细胞相互作用如何影响病毒发病机制。JCPyV 感染高达80%的人群，并在肾脏中建立终身、无症状的持续感染 健康的个体。然而，在免疫功能低下的个体中，JCPyV可以传播到中枢神经系统。 中枢神经系统（CNS），并引起神经胶质细胞的溶解性感染，导致致命的进行性脱髓鞘疾病 多灶性白质脑病（PML）。大约5%的艾滋病毒感染者会发展为PML，一种晚期艾滋病， 定义疾病，以及接受多发性硬化症等疾病免疫调节治疗的个人 患PML的风险更高。越来越多的人接受免疫调节 由于免疫介导的疾病导致的治疗导致PML病例在过去十年中不断增加。超过 共有700例PML病例由单一药物类别引起。PML可能是致命的，特别是当潜在的 免疫抑制是未经治疗的，目前还没有批准的治疗这种毁灭性的 疾病缺乏动物模型和有限的细胞培养模型在很大程度上限制了JCPyV的研究， 单一转化细胞培养模型。最近发表的研究表明，原代细胞培养 模型是体内疾病发病机制的更好代表，因此我们开发了创新的 原代细胞培养模型，以确定JCPyV感染所需的细胞因子。两个综合具体 本研究项目的目的是：1）表征介导JCPyV进入的细胞因子， 感染原代细胞类型，和2）阐明持久性和裂解性JCPyV的细胞类型依赖性差异 感染.这项研究将增强我们对病毒-宿主细胞相互作用如何影响疾病的理解 结果，并将作为一个平台，为抗病毒治疗的发展。通过这项工作，细胞 JCPyV进入原代细胞类型所需的因子将通过功能的丧失和获得来表征 使用基于细胞的测定的方法。通过RNA测序分析的转录组谱分析将用于 确定JCPyV感染如何以细胞类型依赖性方式改变基因表达，以确定途径 与JCPyV发病机制和致死性疾病结局相关。这种组合方法利用了新的 开发了JCPyV感染的原代细胞模型，并利用创新的高通量分析 病毒和细胞蛋白质表达和RNA测序方法。这项研究将填补我们的关键空白， JCPyV生物学知识，并可阐明新的抗病毒靶点或为实验应用提供理论基础 用于预防或治疗致命性疾病PML的市场上疗法。从这项研究中获得的结果也将 为其他致病病毒的进入和信号网络提供更广泛的见解，如冠状病毒， 并提高我们对其他疾病（如癌症）中细胞信号失调的理解。

CELLULAR REPROGRAMMING IN PERSISTENT VS. LYTIC VIRAL INFECTIONS PROJECT SUMMARY The long-range goal of the proposed research is to define the cellular factors that mediate JC polyomavirus (JCPyV) infection to better understand how virus-host cell interactions influence viral pathogenesis. JCPyV infects up to 80% of the population and establishes a lifelong, asymptomatic persistent infection in the kidneys of healthy individuals. However, in immunocompromised individuals JCPyV can spread to the central nervous system (CNS) and cause a lytic infection in glial cells resulting in the fatal, demyelinating disease progressive multifocal leukoencephalopathy (PML). Approximately 5% of individuals with HIV develop PML, a terminal AIDS- defining illness, and individuals receiving immunomodulatory therapies for diseases including multiple sclerosis are at heightened risk for PML development. The increasing number of individuals receiving immunomodulatory therapies due to immune-mediated diseases has led to a rising number of PML cases in the past decade. Over 700 total cases of PML are due to a single drug class. PML can be fatal, especially when underlying immunosuppression is left untreated, and there are currently no approved treatments for this devastating disease. The lack of an animal model and limited cell culture models have largely restricted studies of JCPyV to a single transformed cell culture model. Recently-published studies have revealed that primary cell culture models are a better representation of disease pathogenesis in vivo, and thus we have developed innovative primary cell culture models to define cellular factors that are required for JCPyV infection. Two integrated specific aims are proposed in this research project to: 1) characterize cellular factors that mediate JCPyV entry and infection in primary cell types, and 2) elucidate cell-type dependent differences in persistent and lytic JCPyV infections. This research will enhance our understanding of how virus-host cell interactions influence disease outcomes and will serve as a platform for the development of antiviral treatments. Through this work, cellular factors required for JCPyV entry in primary cell types will be characterized through loss- and gain-of-function approaches using cell-based assays. Transcriptome profiling through RNA sequencing analysis will be used to determine how JCPyV infection alters gene expression in a cell-type dependent manner to identify pathways relevant to JCPyV pathogenesis and fatal disease outcomes. This combinatorial approach utilizes newly developed primary cell models of JCPyV infection and takes advantage of innovative high-throughput analysis of viral and cellular protein expression and RNA sequencing approaches. This research will fill key gaps in our knowledge of JCPyV biology and could elucidate novel antiviral targets or provide rationale for experimental use of on-market therapies to prevent or treat the fatal disease PML. Findings obtained from this research will also provide broader insights into the entry and signaling networks for other pathogenic viruses, like coronaviruses, and improve our understanding of dysregulation of cellular signaling in other diseases such as cancer.