Functional characterization of the enteroviral degradome

肠道病毒降解组的功能表征

• 批准号: 10661758
• 负责人: Mohsan Saeed
• 金额: $ 51.08万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-07 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10661758
• 关键词: Biological Assay; Biology; Cell Culture Techniques; Cell physiology; Cells; Cultured Cells; Development; Disease; Encephalitis; Enterovirus; Enterovirus Infections; Environment; Epithelial Cells; Eukaryotic Initiation Factors; Foundations; Gastroenteritis; Genes; Goals; Human; Human Cell Line; Immune; Immune signaling; Immunologic Monitoring; Impairment; Infection; Innate Immune Response; Integration Host Factors; Interferons; Investigation; Knock-out; Label; Learning; Life Cycle Stages; Maps; Mediating; Mesenchyme; Metabolism; Methodology; Mission; Molecular; Monitor; Myocarditis; N-terminal; Natural Immunity; Normal Cell; Organ; Pathogenesis; Pathway interactions; Peptide Hydrolases; Pneumonia; Predisposition; Preventive; Process; Proteins; Proteolysis; Publications; RNA Splicing; RNA Viruses; Reagent; Regulation; Research; Role; Shapes; Signal Transduction; System; Testing; Therapeutic; Translations; United States National Institutes of Health; Variant; Viral; Viral Physiology; Virus; Virus Assembly; Virus Diseases; Virus Replication; Work; candidate identification; cell type; chromatin remodeling; clinically relevant; design; human disease; human pathogen; induced pluripotent stem cell; innate immune function; innate immune pathways; innovation; insight; intestinal epithelium; novel; novel therapeutic intervention; peroxisome; programs; protein transport; real time monitoring; response; tool; treatment strategy; virus host interaction

Project Summary Enteroviruses are human pathogens that replicate in multiple organs and cause a variety of diseases, including gastroenteritis, pneumonia, myocarditis, and encephalitis. Currently, little is known about how enteroviruses alter the biology of infected cells. This proposed project, which is founded on our recent publication documenting the cellular proteins that enteroviral proteases target for cleavage, will elucidate the role of enteroviral proteases in changing the host cell enviornment during infection. The initial investigations of a subset of cleavage targets have so far yielded two candidate proteins, LSM14A and IRF2BP2, which activated innate immune signaling and inhibited viral replication in cultured cells. Our mechanistic studies have begun to reveal the molecular details of how these proteins limit virus infection and whether their innate immune functions are required for viral inhibition. These preliminary findings have built the foundation to characterize other host targets and determine their ability to regulate enterovirus infection. We have established a toolkit of novel methodologies and reagents that will enable the proposed work. For example, we have implemented an innovative experimental system that consists of human induced pluripotent stem cell-derived mesenchyme-free intestinal epithelial cells, which are highly susceptible to enteroviruses and respond to infection by triggering a robust innate immune program. Similarly, we have generated human cell lines containing interferon-stimulated genes tagged at their endogenous loci with fluorescent proteins, allowing real-time monitoring of immune induction in response to viral infections. We will employ these and other tools to investigate the molecular details of how the host target LSM14A activates innate immune signaling and inhibit viral infections (Aim 1); dissect the function of IRF2BP2 in the enterovirus lifecycle (Aim 2); and generate a comprehensive list of functionally important target proteins that have roles in enterovirus infection and/or antiviral innate immunity (Aim 3). Completion of these aims will reveal host restriction factors against enteroviral replication as well as their mechanisms of action. More broadly, this project will advance the understanding of enteroviral pathogenesis and potentially inform the design of broadly acting preventive and therapeutic strategies against enteroviral infections.

项目摘要 肠病毒是在多个器官中复制并引起多种疾病的人类病原体， 包括胃肠炎，肺炎，心肌炎和脑炎。目前，关于如何 肠病毒改变了感染细胞的生物学。这个拟议的项目，建立在我们最近的 出版物记录肠道病毒蛋白质靶向裂解的细胞蛋白，将阐明 肠病毒蛋白酶在感染过程中改变宿主细胞环境中的作用。 到目前为止，对切割靶标的一部分的初步研究已经产生了两种候选蛋白： LSM14A和IRF2BP2，它们激活先天免疫信号并抑制培养的病毒复制 细胞。我们的机械研究已经开始揭示这些蛋白如何限制病毒的分子细节 感染以及病毒抑制是否需要其先天免疫功能。这些初步 调查结果为表征其他宿主目标并确定其调节能力奠定了基础 肠病毒感染。 我们已经建立了一个新型方法和试剂的工具包，这些工具包将实现拟议的工作。 例如，我们实施了由人类诱导的创新实验系统 多能干细胞衍生的无间隙肠上皮细胞，它们非常容易受到 肠病毒并通过触发强大的先天免疫程序来应对感染。同样，我们也有 产生的人类细胞系，其中包含在其内源性基因座标记的干扰素刺激的基因 荧光蛋白，允许对病毒感染的反应实时监测免疫诱导。 我们将使用这些和其他工具来研究主机目标LSM14A的分子细节 激活先天的免疫信号传导并抑制病毒感染（AIM 1）；剖析IRF2BP2的功能 肠病毒生命周期（AIM 2）；并产生全面的功能重要目标列表 在肠病毒感染和/或抗病毒先天免疫中具有作用的蛋白质（AIM 3）。 这些目的的完成将揭示针对肠病毒复制及其的宿主限制因素 作用机理。更广泛地，该项目将提高对肠病毒的理解 发病机理并有可能告知广泛作用的预防和治疗策略的设计 反对肠病毒感染。