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

研究项目2（RP 2）：项目总结/摘要 这个整体计划项目旨在解决有关埃博拉病毒（EBOV）的重大知识差距：如何 感染导致致病性而不是保护性免疫应答。在这个项目中，重点是如何EBOV 感染改变了关键免疫细胞类型和严重免疫缺陷细胞中的转录后基因调控。 感染的（例如，肝细胞）。关于EBOV感染如何影响宿主基因表达的理解有限 在转录后水平。事实上，尽管基因转录后调控的重要性 对于多种致病性人类病毒的表达是明显的，这种水平的调节已经被充分研究， 丝状病毒 研究项目2（RP 2）的中心假设是细胞类型特异性和基因特异性转录后 EBOV疾病中的免疫应答导致免疫细胞的适应不良表型，最终导致 “免疫麻痹”和炎症过度。为了解决这个假设，我们将描述 EBOV感染期间原代人免疫和非免疫细胞的转录后景观。我们将 EBOV感染对宿主RNA加工和翻译的影响。大规模平行测序 蛋白质基因组学核心（核心C）使之成为可能的方法将用于定义RNA加工 事件，RNA修饰和mRNA翻译效率在幼稚和感染细胞中的转录组范围内。 重要的是，所有三个实验都将使用BSL-4核心（核心B）中相同实验产生的样本 因此，我们计划将我们的数据与RP 1和RP 3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（核心D）合作进行。这些 实验将产生新的见解EBOV感染对相关细胞中细胞基因表达的影响 类型我们还将描述EBOV在非人类中体内感染期间的转录后景观。 灵长类动物使用这些实验的数据，我们将用核心D构建一个全面的模型， 在不同的免疫细胞类型中调节EBOV感染的调节宿主途径。该模型将识别 候选基因代表了这些途径的重要节点，因此是EBOV的潜在弱点。 与核心B合作，该项目将验证转录后改变的基因和事件，并询问 它们对EBOV感染的细胞类型特异性应答是否在功能上重要。 该项目的完成将揭示宿主对EBOV感染的新的和未被充分认识的反应层， 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发病机制。此外，我们将确定关键的新宿主和/或病毒靶点， 破坏这种致命病毒的致病性

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.