Analysis of cellular factors limiting productive JC virus infections

限制生产性 JC 病毒感染的细胞因素分析

• 批准号: 10312804
• 负责人: JAMES M PIPAS
• 金额: $ 19.69万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-07 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10312804
• 关键词: Acquired Immunodeficiency Syndrome; Affect; Antiviral Response; BK Virus; Blood; Brain; Cell Death; Cells; Computer Analysis; Critical Pathways; Cystitis; Disease; Encephalopathies; Epithelial Cells; Equilibrium; Family; Gene Expression; Genes; Genetic; Growth; Hemorrhage; Human; IRF3 gene; Immune response; Immunocompromised Host; Immunologic Techniques; Immunosuppression; Individual; Infection; Inflammatory Response; Interferon Activation; Interferons; JC Virus; Kidney; Kidney Diseases; Link; Maintenance; Measures; Molecular; Monitor; Nerve Degeneration; Neuraxis; Outcome; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Play; Polyomavirus; Population; Production; Proteins; Proximal Kidney Tubules; Research; Role; STAT1 gene; Signal Pathway; System; Tissues; Urine; Viral; Viral Load result; Virion; Virus; Virus Diseases; Virus Replication; Virus-Cell Membrane Interaction; cell type; chronic infection; clinically relevant; cytokine; established cell line; experimental study; inhibitor; kidney cell; knock-down; member; monolayer; organ transplant recipient; particle; prevent; response; single-cell RNA sequencing; transcriptomics

Project Summary JCV and BKV are members of the Polyomavirus family that infect a large portion of the human population. Under normal conditions both viruses establish a lifelong, asymptomatic, persistent infection in the kidneys, and are frequently detected in urine and circulating blood without apparent consequences. While in most cases JCV and BKV are harmless, they can reactivate in immunosuppressed patients, and then induce serious diseases. However, the outcome from either infection is very different: BKV infection induces an inflammatory response and tissue damage in the kidney that results in nephropathy and hemorrhagic cystitis, while JCV reactivation causes fatal neurodegeneration in the brain, but only a few nephropathy cases. The factors governing the equilibrium between viral productive infection and persistent infection are unknown, as are the mechanisms controlling the different cellular responses to similar polyomaviruses. This application focuses on understanding the basis for cellular responses to JCV in a relevant primary human cell type, renal proximal tubule epithelial cells (RPTE). In contrast with BKV, whose infection induces extensive cell death around 5 dpi and the release of about 40 infectious particles/cell, JCV establishes a low-level infection in RPTE, causing minimal cytopathic effects and persisting for at least 3 weeks without inducing a full productive infection. Inoculation of RPTE with JCV results in induction of the interferon response, which could prevent viral expansion. This research will use human primary cells to identify the factors that influence infection outcomes: productive or persistent. Activation of the interferon response by JCV infection in RPTE will be assessed using both molecular and functional approaches to identify components of the pathway that are critical for responding to the virus and for limiting viral infection. Then, single cell transcriptomics will be applied to RPTE to assess the response of cell subpopulations to infection and to identify genes that play a role in restricting JCV replication. Finally, a comparison between the responses observed upon infection of RPTE with either BKV or JCV will be used to identify cellular genes that distinguish abortively versus productively infected cells. This application seeks to understand the unique responses to JCV in a clinically relevant human primary cell from the kidneys, the mechanisms that drive those responses and how the same cells respond in a different fashion to infection with BKV. This research will increase our understanding of principles governing cellular responses to viral infection, and how these responses influence the outcome of infection.

项目摘要 JCV和BKV是感染大部分人口的多瘤病毒家族的成员。 在正常情况下 并经常在尿液和循环血液中检测到而没有明显的后果。而在大多数情况下 JCV和BKV是无害的，可以在免疫抑制的患者中重新激活，然后诱发严重 疾病。但是，两种感染的结果都大不相同：BKV感染引起炎症 肾脏的反应和组织损伤导致肾病和出血性膀胱炎，而JCV 重新激活会导致大脑致命的神经退行性变化，但仅少数肾病病例。因素 管理病毒生产感染和持续感染之间的平衡是未知的， 控制对类似多瘤病毒的不同细胞反应的机制。 该应用的重点是理解相关主要人类中对JCV的细胞响应的基础 细胞类型，肾近端小管上皮细胞（RPTE）。与BKV相反，BKV的感染诱导了广泛 细胞死亡约5 dpi，释放约40个传染性颗粒/细胞，JCV建立了低级 在RPTE中感染，导致最小的细胞质效应，并持续至少3周 生产感染。用JCV接种RPTE会导致干扰素反应诱导，这可能 防止病毒膨胀。 这项研究将使用人类原代细胞来确定影响感染结果的因素：生产性 或持久。 JCV感染在RPTE中激活干扰素反应将使用两者进行评估 分子和功能性方法，以识别途径的组成部分，这对于对响应至关重要 病毒和限制病毒感染。然后，将将单细胞转录组学应用于RPTE来评估 细胞亚群对感染的反应并确定在限制JCV复制中起作用的基因。 最后，将RPTE感染在用BKV或JCV感染时观察到的反应之间的比较将是 用于鉴定区分堕胎与有效感染细胞的细胞基因。 该应用程序旨在了解临床相关的人类原代细胞中对JCV的独特响应 从肾脏中，驱动这些响应的机制以及相同细胞的响应方式不同 BKV感染的时尚。这项研究将增加我们对控制蜂窝的原则的理解 对病毒感染的反应，以及这些反应如何影响感染的结果。