Role of IRE1 Alpha in Coronavirus Infections

IRE1 Alpha 在冠状病毒感染中的作用

• 批准号: 10590642
• 负责人: Susan Leilani Fink
• 金额: $ 54.47万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-11 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10590642
• 关键词: 2019-nCoV; Affect; Aging; Anti-Inflammatory Agents; Biogenesis; COVID-19; COVID-19 pathogenesis; COVID-19 patient; COVID-19 risk; Cell Culture Techniques; Cell physiology; Clinical; Coronavirus; Coronavirus Infections; Data; Diabetes Mellitus; Disease; Endoplasmic Reticulum; Etiology; Evaluation; Floods; Future; Genetic; Human; Hypertension; Infection; Inflammation; Inflammatory; Inflammatory Response; Inflammatory Response Pathway; Integration Host Factors; Investigation; Life Cycle Stages; Mediating; Medical; Membrane Fusion; Messenger RNA; Modeling; Molecular; Molecular Virology; Obesity; Pathogenesis; Pathogenicity; Pathway interactions; Pharmaceutical Preparations; Play; Prediction of Response to Therapy; Process; Production; Prognostic Marker; Protein Biosynthesis; Proteins; RNA; RNA Splicing; RNA replication; Risk Factors; Role; SARS-CoV-2 infection; Severity of illness; Specificity; Specimen; Testing; Therapeutic Effect; Viral; Virion; Virus Diseases; Virus Replication; Work; XBP1 gene; biological adaptation to stress; cell type; chemical genetics; comorbidity; current pandemic; cytokine; endoplasmic reticulum stress; experience; experimental study; gene induction; genetic approach; human coronavirus; human disease; in vivo; inhibitor; insight; mouse model; novel; nuclease; pandemic virus; pre-clinical; response; severe COVID-19; therapeutic target; tool; transcription factor; virology

Project Summary Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a recently emergent, currently pandemic virus and etiological agent of Coronavirus Induced Disease-19 (COVID-19). Despite a flood of scientific investigation, critical gaps remain in our understanding of the basic cellular processes that facilitate replication of coronaviruses, including SARS-CoV-2, and contribute to the pathogenesis of severe disease. Our preliminary data demonstrate that IRE1α, a component of the cellular response to endoplasmic reticulum (ER) stress, is required for SARS-CoV-2 replication and inflammatory cytokine responses. However, the stage(s) of the viral life cycle and downstream cellular pathways that mediate these effects remain completely unknown. ER stress and IRE1α activation are well-associated with conditions such as obesity, diabetes, hypertension, and aging, all of which are risk factors for severe manifestations of COVID-19. We hypothesize that comorbidity-associated ER stress primes both exuberant viral replication and pathogenic inflammatory cytokine production via IRE1α. This project leverages our unique ability to test this hypothesis using cell culture infection models, as well as specimens from patients with COVID-19. IRE1α inhibitors are under evaluation for treatment of non-infectious human diseases, and we propose that this project will provide preclinical evidence for the novel application of these drugs to treat infections with SARS-CoV-2 and potentially other human coronaviruses. The experiments outlined in this proposal will determine the molecular mechanism(s) by which IRE1α supports SARS-CoV-2 infection. IRE1α is a nuclease which initiates nonconventional splicing of XBP1 mRNA, which encodes a pleiotropic transcription factor. IRE1α also targets other specific RNAs leading to their degradation. We will determine whether the requirement for IRE1α is XBP1-dependent or -independent and dissect the downstream cellular processes that facilitate SARS-CoV-2 replication and inflammatory cytokine responses (Aim 1). We will systematically identify stage(s) of the SARS-CoV-2 life cycle that require IRE1α (Aim 2). We predict that IRE1α most likely supports biogenesis of ER-derived viral replication platforms, and will focus experiments on this hypothesis. We predict that IRE1α represents a target for dual anti-viral and anti- inflammatory therapy and will test this in mouse models of SARS-CoV-2 infection (Aim 3). Finally, we will determine whether IRE1α activation occurs during human infection and ER stress is a prognostic marker for severe COVID-19. Together, the results of this project will reveal basic cellular processes occurring during coronavirus infection and host factors critical for the pathogenesis of COVID-19. .

项目摘要 严重的急性呼吸综合症冠状病毒2（SARS-COV-2）是最近出现的，目前大流行 冠状病毒诱导疾病19（COVID-19）的病毒和病因。尽管科学泛滥 调查，我们对促进复制的基本细胞过程的理解仍然存在关键差距 包括SARS-COV-2在内的冠状病毒，并有助于严重疾病的发病机理。 我们的初步数据表明，IRE1α是内质网的细胞反应的组成部分 （ER）SARS-COV-2复制和炎症细胞因子反应所必需的应力。但是， 病毒生命周期的阶段和介导这些作用的下游细胞途径保持不变 未知。 ER应激和IRE1α激活与肥胖，糖尿病， 高血压和衰老，所有这些都是Covid-19的严重表现的危险因素。我们假设 合并症相关的ER应力素数既激增病毒复制和致病性炎症 通过IRE1α产生细胞因子。该项目利用了我们使用单元格检验该假设的独特能力 培养感染模型以及Covid-19患者的标本。 IRE1α抑制剂不在 评估非感染人类疾病的治疗，我们建议该项目提供 这些药物在治疗SARS-COV-2感染的新颖应用的临床前证据，并有可能 其他人类冠状病毒。 该提案中概述的实验将确定IRE1α支持的分子机制 SARS-COV-2感染。 IRE1α是一种核酸酶，启动了XBP1 mRNA的非惯性剪接，该剪接 编码多效性转录因子。 IRE1α还靶向其他特定的RNA导致其降解。 我们将确定IRE1α的需求是XBP1依赖性还是非依赖性并剖析 促进SARS-COV-2复制和炎症细胞因子反应的下游细胞过程 （目标1）。我们将系统地确定需要IRE1α的SARS-COV-2生命周期的阶段（AIM 2）。我们 预测IRE1α最有可能支持ER衍生的病毒复制平台的生物发生，并将集中 关于此假设的实验。我们预测IRE1α代表双病毒和抗抗病毒的目标 炎症疗法将在SARS-COV-2感染的小鼠模型中测试（AIM 3）。最后，我们会的 确定IRE1α激活是否发生在人类感染期间，而ER应激是预后的标志物 严重的Covid-19。总之，该项目的结果将揭示在此期间发生的基本蜂窝过程 冠状病毒感染和宿主因子对19009的发病机理至关重要。 。