Research Project 2: Role of Posttranscriptional Regulatory Networks in the Pathogenesis of Ebola Virus Disease

研究项目2：转录后调控网络在埃博拉病毒疾病发病机制中的作用

• 批准号: 10394321
• 负责人: Mariano A. Garcia-Blanco
• 金额: $ 36.12万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10394321
• 关键词: Address; Affect; Antisense Oligonucleotides; Area; Bioinformatics; Candidate Disease Gene; Cell physiology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Data; Data Analyses; Dendritic Cells; Development; Ebola Hemorrhagic Fever; Ebola virus; Endothelial Cells; Event; Filovirus; Gene Expression; Gene Expression Regulation; Genes; Genetic Translation; Hepatitis C; Hepatocyte; Human; Immune; Immune Tolerance; Immune response; Investigation; Knowledge; Literature; Massive Parallel Sequencing; Mediating; Methods; Modeling; Modification; Molecular; Pathogenesis; Pathogenicity; Pathway interactions; Phenotype; Post-Transcriptional Regulation; Primary Infection; Process; Protein Isoforms; Publishing; RNA; RNA Interference; RNA Processing; RNA Splicing; RNA-Binding Proteins; Regulation; Regulator Genes; Research Project Grants; Role; Sampling; T cell response; T-Lymphocyte; Testing; Tissues; Translations; Viral; Viral Pathogenesis; Virus; Virus Diseases; Virus Replication; Work; cell type; data resource; experimental study; exposed human population; in vivo; insight; kidney cell; knock-down; knowledge integration; monocyte; nonhuman primate; novel; programs; proteogenomics; response; therapeutic RNA; transcriptome; virtual

RESEARCH PROJECT 2 (RP2): PROJECT SUMMARY/ABSTRACT This overall Program Project seeks to address a significant knowledge gap concerning Ebola virus (EBOV): how infection leads to a pathogenic rather than protective immune response. In this project, the focus is on how EBOV infection alters posttranscriptional gene regulation in critical immune cell types and in cells that are heavily infected (e.g., hepatocytes). There is limited understanding of how EBOV infection impacts host gene expression at the posttranscriptional level. Indeed, although the importance of posttranscriptional regulation in gene expression for multiple pathogenic human viruses is evident, this level of regulation has been understudied for filoviruses. The central hypothesis of Research Project 2 (RP2) is that cell type-specific and gene-specific posttranscriptional responses in EBOV disease contribute to the maladaptive phenotypes of immune cells, eventually leading to “immune paralysis” and hyperinflammation. To address this hypothesis, we will characterize the posttranscriptional landscape during EBOV infections of primary human immune and nonimmune cells. We will decipher the influence of EBOV infection on host RNA processing and translation. Massively parallel sequencing approaches made possible by the Proteogenomics Core (Core C) will be used to define RNA processing events, RNA modifications, and mRNA translation efficiency transcriptome-wide in naïve and infected cells. Importantly, samples generated from the same experiments in the BSL-4 Core (Core B) will be used by all three projects and thus we plan to compare our data with that of RP1 and RP3. Data analysis, integration, and modelling will be performed in collaboration with the Bioinformatics and Modeling Core (Core D). These experiments will yield new insights into the effect of EBOV infection on cellular gene expression in relevant cell types. We will also characterize the posttranscriptional landscape during EBOV infection in vivo in nonhuman primates. Using the data from these experiments, with Core D we will construct a comprehensive model of regulated host pathways that modulate EBOV infection in different immune cell types. This model will identify candidate genes that represent important nodes for these pathways and thus potential vulnerabilities for EBOV. In collaboration with Core B, this project will validate posttranscriptionally altered genes and events, and ask whether they are functionally important to cell-type specific responses to EBOV infection. Completion of this project will reveal new and underappreciated layers of host responses to EBOV infection, and provide a rich data resource for the field that will lead to new hypotheses to address the molecular basis for EBOV pathogenesis. Moreover, we will identify critical new host and/or viral targets that can be exploited to disrupt the pathogenicity of this deadly virus.

研究项目2（RP2）：项目摘要/摘要 这个整体计划项目旨在解决有关埃博拉病毒（EBOV）的重要知识差距：如何 感染导致致病性，而不是保护性免疫反应。在这个项目中，重点是eBov 感染改变了临界免疫细胞类型的转录后基因调节，在大量细胞中调节 感染（例如，肝细胞）。对EBOV感染如何影响宿主基因表达的理解有限 在转录后级别。确实，尽管后期调节在基因中的重要性 多种致病性人类病毒的表达是证据，这种调节水平已被理解为 丝状病毒。 研究项目2（RP2）的中心假设是细胞类型特异性和基因特异性转录后 EBOV疾病中的反应导致免疫小球的适应不良表型，最终导致 “免疫瘫痪”和过度炎症。为了解决这一假设，我们将表征 初次人免疫和非免疫细胞EBOV感染期间的转录后景观。我们将 破译EBOV感染对宿主RNA处理和翻译的影响。大量平行测序 蛋白质组学核心（核心C）使方法成为可能的方法将用于定义RNA处理 幼稚和感染细胞中的事件，RNA修饰和mRNA翻译效率在整个转录组中。 重要的是，将从BSL-4核心（核心B）中同一实验产生的样品均应均由所有三个 项目，因此我们计划将我们的数据与RP1和RP3的数据进行比较。数据分析，集成和 建模将与生物信息学和建模核心（Core D）合作进行。这些 实验将产生对EBOV感染对相关细胞中细胞基因表达的影响的新见解 类型。我们还将表征非人类的EBOV感染期间的转录后景观 使用来自这些实验的数据，使用核心D，我们将构建一个综合模型 调节的宿主途径可调节不同免疫细胞类型的EBOV感染。该模型将确定 代表这些途径重要节点的候选基因，因此是EBOV的潜在漏洞。 与Core B合作，该项目将验证转录后改变的基因和事件，并询问 它们是否对细胞类型对EBOV感染的特异性反应在功能上很重要。 该项目的完成将揭示新的和低估的宿主对EBOV感染的反应，以及 为该领域提供丰富的数据资源，这将导致新的假设以解决分子基础 EBOV发病机理。此外，我们将确定可以探索的关键新宿主和/或病毒目标 破坏这种致命病毒的致病性。