Research Project 3: Role of Posttranslational Protein Modifications in the Pathogenesis of Ebola Virus Disease

研究项目3：翻译后蛋白修饰在埃博拉病毒疾病发病机制中的作用

基本信息

批准号：
10188761
负责人：
Ricardo Rajsbaum
金额：
$ 34.52万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
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/10188761
关键词：
Address Affect Africa Animal Model Antibody Therapy Antiviral Agents Antiviral Response Bioinformatics Biological Assay Cells Cessation of life Co-Immunoprecipitations Collaborations Complex Data Dendritic Cells Deubiquitination Development Disease Ebola Hemorrhagic Fever Ebola virus Endothelial Cells Enzymes Epidemic Equilibrium Funding Gene Expression Genetic Transcription Goals Hepatocyte Human Immune Immune response Immune signaling Immunological Models In Vitro Infection Inflammation Inflammatory Inflammatory Response Injury Interferons Knock-out Knowledge Laboratories Ligase Link Liver Mass Spectrum Analysis Mediating Modeling Modification Molecular Molecular Target Monoclonal Antibodies Outcome Outcome Study Pathogenesis Pathogenicity Pathway interactions Pattern Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Physiological Post-Translational Protein Processing Post-Translational Regulation Primary Infection Process Production Protein Dephosphorylation Proteomics Regulation Research Project Grants Role Sampling Signal Pathway Signal Transduction System T-Lymphocyte Ubiquitin Ubiquitination Validation Vascular Endothelium Viral Viral Proteins Virus Diseases Virus Replication Work cell type cytokine cytokine release syndrome in vivo insight kidney cell kidney vascular structure knock-down monocyte mutant nonhuman primate novel strategies overexpression pathogenic virus proteogenomics response transforming virus ubiquitin ligase virus core

项目摘要

RESEARCH PROJECT 3 (RP3): PROJECT SUMMARY/ABSTRACT The epidemic of Ebola virus (EBOV) in Africa in 2013-2016 and its reappearance in subsequent years have reached alarming numbers of infections and deaths, prompting extensive efforts to develop antiviral strategies; however, none have thus far been licensed for use in humans. Pathogenesis of EBOV infection derives from a combination of the viral suppression of the host's antiviral protective immune responses and induction of hyper- inflammation, including a “cytokine storm”. Cytokine production and inflammatory responses require activation of multiple signaling pathways that are extensively regulated at the posttranslational level to maintain a balance between efficient antiviral responses and excessive inflammation. Posttranslational modifications (PTMs) of pro- teins serve as molecular regulatory mechanisms, a process which requires the activity of specific enzymes (i.e., kinases, ubiquitin ligases, etc.). Thus, the expression and activation patterns of these enzymes in specific cell types determines the complex regulation that balances an effective immune physiological response. The goal of Research Project 3 (RP3) is to elucidate how EBOV transforms the PTM cell-type specific landscape leading to immune dysregulation and disease, with the long-term goal of applying this knowledge in the development of effective approaches to treat the disease caused by EBOV. Our preliminary data indicate that interferon antagonist domains of EBOV proteins can hijack the host ubiquitin system to increase virus replication while causing cell-type specific dysregulation of signaling pathways that control hyperinflammation. However, it is not known which PTM enzymes or what changes in host protein mod- ifications drive immune dysregulation and disease. Our hypothesis is that EBOV targets specific enzymes that regulate PTM in a cell-type specific manner resulting in uncontrolled inflammation, while also blocking protective innate immune signaling. We will use novel approaches to assess how EBOV infection dysregulates immune signaling by targeting unconventional PTMs, in vitro and in vivo. With information obtained from the BSL-4 Core (Core B), Proteogenomics Core (Core C), and Bioinformatics and Modeling Core (Core D), and from RP1 and RP2 we will build a complete model of inflammatory pathways activated during EBOV infection. In Aim 1 we will characterize the PTM landscape during EBOV infection in primary human immune and nonimmune cells, and in Aim 2 in cells isolated from infected nonhuman primates, using mass spectrometry analysis and validation assays. In Aim 3, we will validate PTM enzymes and their modifications, and elucidate cell-type specific PTM- mediated mechanisms that result in dysregulated immune responses to EBOV. The outcome of these studies is significant because it will provide fundamental knowledge on the function of specific PTMs during infection and uncover molecular targets to treat pathogenic inflammation. Our work on posttranslational regulation together with information derived from changes in the transcriptional (RP1) and post- transcriptional (RP2) landscapes will provide a comprehensive model of immune dysregulation.

研究项目3（RP3）：项目摘要/摘要 2013 - 2016年非洲埃博拉病毒（EBOV）的流行及其在随后几年的重新出现达到了令人震惊的感染和死亡人数，促使人们为制定抗病毒策略而进行了广泛的努力；但是，到目前为止，没有人获得许可用于人类。 EBOV感染的发病机理源自A 宿主抗病毒保护免疫复杂的病毒抑制和诱导的组合炎症，包括“细胞因子风暴”。细胞因子的产生和炎症反应需要激活在翻译后水平上受到广泛调节以保持平衡的多个信号通路在有效的抗病毒反应和过度炎症之间。 pro的翻译后修饰（PTMS） Teins是分子调节机制，该过程需要特定酶的活性（即激酶，泛素连接酶等）。那就是这些酶在特定细胞中的表达和激活模式类型决定了平衡有效免疫物理反应的复杂调节。目标研究项目3（RP3）是为了阐明EBOV如何改变PTM细胞类型的特定景观领先免疫失调和疾病，其长期目标是将这种知识应用于发展有效治疗由EBOV引起的疾病的方法。我们的初步数据表明EBOV蛋白的干扰素拮抗剂域可以劫持宿主泛素增加病毒复制的系统，同时引起细胞类型的信号传导途径的特异性功能失调控制高炎症。但是，尚不清楚哪种PTM酶或宿主蛋白模式的变化是什么影响免疫失调和疾病。我们的假设是EBOV靶向特定酶以细胞类型的特异性方式调节PTM，导致不受控制的注射，同时也阻止了受保护的PTM 先天免疫信号传导。我们将使用新颖的方法评估EBOV感染如何失调免疫通过靶向非常规PTM的体外和体内信号传导。从BSL-4核心获得的信息（核B），蛋白质组学核心（核心C），生物信息学和建模核心（核心D），以及RP1 RP2我们将建立一个在EBOV感染期间激活的炎症途径的完整模型。在目标1中我们将在EBOV感染期间特征PTM景观中的原代人免疫和非免疫细胞，并使用质谱分析和验证从被感染的非人类隐私分离的细胞中的AIM 2中测定。在AIM 3中，我们将验证PTM酶及其修饰，并阐明细胞类型的特异性PTM- 导致对EBOV的免疫回报失调的介导机制。这些研究的结果很重要，因为它将提供有关功能的基本知识感染过程中的特定PTM和发现病原感染的分子靶标。我们的工作翻译后调节以及从转录（RP1）和后变化得出的信息转录（RP2）景观将提供全面的免疫失调模型。