Research Project 1: Role of Epigenetic and Transcriptional Mechanisms in the Pathogenesis of Ebola Virus Disease

研究项目1：表观遗传和转录机制在埃博拉病毒疾病发病机制中的作用

• 批准号: 10602491
• 负责人: Alexander Bukreyev
• 金额: $ 41.18万
• 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/10602491
• 关键词: 2019-nCoV; ATAC-seq; Acute; Address; Animals; Antiinflammatory Effect; Apoptosis; Area; B-Lymphocytes; Bioinformatics; Cell physiology; Cells; Cessation of life; ChIP-seq; Chemicals; Chromatin; Collaborations; Cytoplasm; Data; Data Analyses; Dendritic Cells; Deposition; Development; Disease; Ebola; Ebola Hemorrhagic Fever; Ebola virus; Ebola virus envelope glycoprotein; Endothelial Cells; Enzymes; Epigenetic Process; Equilibrium; Gene Activation; Gene Expression; Genes; Genetic Transcription; Hepatocyte; Human; Immune; Immune Tolerance; Immune response; Immunity; Immunology; In Vitro; Infection; Inflammation; Inflammation Mediators; Inflammatory; Interferon alpha; Interferons; Investigation; Joints; Journals; Knowledge; Lymphopenia; Marburgvirus; Mediating; Methods; Modeling; Molecular; Mus; Myelogenous; Nature; Outcomes Research; Paper; Pathogenesis; Pathogenicity; Pathologic; Physiological; Process; Production; Protein Isoforms; Publishing; Research; Research Personnel; Research Project Grants; Role; Science; Signal Transduction; Site; T cell response; T-Cell Activation; T-Lymphocyte; TLR4 gene; Testing; Topoisomerase; Transcriptional Regulation; Translational Regulation; Treatment outcome; Viral Pathogenesis; Virus; Virus Diseases; Work; cell type; cytokine; cytokine release syndrome; epigenetic regulation; epigenome; gene repression; histone methylation; in vivo; inhibitor; innovation; kidney cell; mRNA Expression; medical schools; monocyte; nonhuman primate; pathogen; pharmacologic; programs; proteogenomics; response; single cell analysis; single cell sequencing; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; virology

RESEARCH PROJECT 1 (RP1): PROJECT SUMMARY/ABSTRACT Pathogenesis of the disease caused by Ebola virus (EBOV) is characterized by a multifaceted and contradictory interplay between the virus and the host. The effect of EBOV infection is characterized by deficient T-cell responses and lymphopenia, which at least in part results from stimulation by virus-infected dendritic cells whose maturation is suppressed by interferon-inhibiting domains of the virus. In addition, EBOV infections are characterized by hyperinflammation, contributed by interaction of the EBOV envelope glycoprotein (GP) with TLR4 on T cells and possibly by other mechanisms, resulting in massive secretion of cytokines. This dysregulated immune response results from disruption of transcriptional networks, many caused by alterations in the epigenome, as evidenced by (1) chemical inhibition of type 1 topoisomerase resulting in reduced expression of proinflammatory mediators induced by EBOV in infected cells, (2) inhibition of TLR4 signaling, which is regulated at epigenetic level, promoting survival of EBOV-infected mice, and (3) treatment of EBOV- infected mice with a global histone methylation inhibitor that upregulates production of IFNα and protects animals from death. The central hypothesis of Research Project 1 (RP1) is that cell-specific and gene-specific regulation of transcription in response to EBOV infection leads to contrasting responses that paradoxically cause a pathogenic immune response and “immune paralysis”. To address this hypothesis, the epigenetic and transcriptional landscape during EBOV infections in vitro and in vivo will be characterized. A comprehensive analysis of the transcriptome alterations and epigenetic changes caused by EBOV infection will be performed in both human primary cells and specific cell types isolated from infected nonhuman primates. Investigations involving the expression of mRNAs that encode transcription factors will be done to determine the deposition of epigenetic marks and their role in gene activation and repression, as well as investigating the effects of chromatin accessibility on expression and secretion of inflammatory mediators using cells from human donors. This project will target key transcriptional nodes identified by modeling EBOV infection. The data generated in vitro and in vivo, together with data from in RP2 and RP3, will be integrated by the Bioinformatics and Modelling Core (Core D) in a model. The transcriptional networks identified in the model will predict critical nodes that represent vulnerabilities that will be targeted to modulate cell response and reduce EBOV pathogenesis. For example, chromatin-resident enzymes and transcription factors that induce expression of genes causing inflammation and other pathological changes will be targeted. The study will result in the characterization of epigenetic and transcriptional responses to EBOV; to date no epigenetic response to any BSL-4 level virus has been characterized. The study will also result in the development of approaches to treat EBOV disease by targeting host transcriptional and epigenetic functions.

研究项目1（RP1）：项目摘要/摘要 由埃博拉病毒（EBOV）引起的疾病的发病机理的特征是多方面和矛盾 病毒与宿主之间的相互作用。 EBOV感染的影响的特征是T细胞不足 反应和淋巴细胞减少症，至少部分是由病毒感染的树突状细胞刺激的结果 病毒的干扰素抑制域抑制成熟。此外，EBOV感染是 以高炎症为特征，由EBOV包膜糖蛋白（GP）的相互作用 TLR4在T细胞上，通过其他机制可能导致细胞因子的大量分泌。这 转录网络中断的免疫反应失调结果，许多因素是由改变引起的 在表观基因组中，（1）1型拓扑异构酶的化学抑制作用证明了 EBOV在感染细胞中诱导的促炎性介质的表达，（2）抑制TLR4信号传导， 在表观遗传水平上受到调节，促进EBOV感染小鼠的存活，以及（3）EBOV- 用全球组蛋白甲基化抑制剂感染小鼠，该抑制剂上调IFNα的产生并保护动物 从死亡。 研究项目1（RP1）的中心假设是细胞特异性和基因特异性调节 响应EBOV感染的转录导致对比反应，这些反应自相矛盾地引起致病性 免疫反应和“免疫麻痹”。为了解决这一假设，表观遗传和转录 EBOV感染期间体外和体内的景观将被表征。对 由EBOV感染引起的转录组改变和表观遗传变化将在两个人中进行 主细胞和特定细胞类型从感染的非人类隐私中分离出来。调查涉及 将进行编码转录因子的mRNA表达以确定表观遗传的沉积 标记及其在基因激活和表达中的作用，并研究染色质的作用 使用人类供体的细胞对炎症介质表达和分泌的可及性。 该项目将针对通过对EBOV感染进行建模的关键转录节点。生成的数据 体外和体内以及来自RP2和RP3中的数据将由生物信息学和 在模型中建模核心（核心D）。模型中确定的转录网络将预测关键 代表漏洞的节点将针对调节细胞响应并减少EBOV 发病。例如，染色质驻留酶和影响的转录因子 引起感染和其他病理变化的基因将成为针对性。 该研究将导致表观遗传和对EBOV的转录反应的表征。迄今为止否 对任何BSL-4水平病毒的表观遗传反应均已表征。该研究还将导致 通过靶向宿主的转录和表观遗传功能来开发治疗EBOV疾病的方法。