Host inflammatory response to SARS-CoV-2

宿主对 SARS-CoV-2 的炎症反应

• 批准号: 10192439
• 负责人: Michael Gale
• 金额: $ 26.48万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10192439
• 关键词: 2019-nCoV; Address; Adult Respiratory Distress Syndrome; Affect; Applications Grants; Autocrine Communication; Bioinformatics; Biological Assay; Biological Markers; COVID-19; COVID-19 intervention; COVID-19 patient; COVID-19 severity; COVID-19 treatment; Cell Death; Cell Line; Cells; Cessation of life; Clustered Regularly Interspaced Short Palindromic Repeats; Common Cold; Complex; Computational Biology; Coronavirus; Coughing; Cytokine Signaling; Disease; Distress; Epithelial Cells; Event; Future; Gene Expression; Gene Expression Profile; Genes; Goals; Headache; Heart Arrest; Human; Human Cell Line; Hyperactivity; IL6 gene; Immune; Immune response; Immunofluorescence Immunologic; Individual; Infection; Inflammation; Inflammation Process; Inflammatory; Inflammatory Response; Inflammatory Response Pathway; Interleukin-1 beta; Kidney Failure; Laboratories; Lung; Maps; Middle East Respiratory Syndrome; Middle East Respiratory Syndrome Coronavirus; Minor; Modality; Molecular; Paracrine Communication; Pathogenicity; Pathway interactions; Patients; Pharmacology; Population; Process; Production; Prognosis; SARS coronavirus; SARS-CoV-2 infection; Severe Acute Respiratory Syndrome; Signal Pathway; Signal Transduction; Sore Throat; Symptoms; TNF gene; Techniques; Tissues; United States; Virus; Virus Diseases; Work; acute infection; base; cytokine; cytokine release syndrome; experience; gastrointestinal; immunoregulation; inhibitor/antagonist; knockout gene; mortality; novel coronavirus; programs; severe COVID-19; single-cell RNA sequencing; transcriptome; transcriptomics; transmission process

Each year, humans encounter a variety of seasonal corona viruses that cause minor symptoms similar to the common cold. In contrast, SARS-CoV-2, which is responsible for COVID-19 (coronavirus disease 2019), has caused more than 110,000 deaths in the United States alone as of June 4, 2020. For unknown reasons, COVID- 19 disease severity is extremely variable, with many infected individuals experiencing mild symptoms including sore throat, headache, and cough or no symptoms at all. However, a significant number of infected individuals experience severe disease, including acute respiratory distress syndrome, renal failure, cardiac arrest, and gastrointestinal distress. Importantly, SARS-CoV-2 infection can trigger a hyperactive immune response that results in a cytokine storm, which is a massive release of pro-inflammatory molecules that can cause fatal tissue damage and likely causes the most severe disease symptoms. What triggers and regulates a cytokine storm is unclear, and studies are needed to reveal the processes by which SARS-CoV-2 induces inflammation, and how SARS-CoV-2 infection imparts COVID19 disease. Thus, the overarching goal of this proposal is to determine how inflammatory responses to SARS-CoV-2 are generated. To address this goal, we propose the following Specific Aims: 1. Define the inflammatory cytokine response induced by SARS-CoV-2. We will evaluate cytokine production from cells infected with SARS-CoV-2 using cell lines derived from lung and immune cells that populate the lung. Additionally, because severe COVID-19 patients present with elevated levels of the cytokine IL-1β and because IL-1β is released following the inflammatory form of cell death known as pyroptosis, we will assess the ability of SARS-CoV-2 to stimulate pyroptosis. We will then use CRISPR-based gene knockout and pharmacologic inhibitors to map the molecular pathways required for production and release of the observed cytokines. 2. Determine the inflammatory transcriptome across infected and bystander cells in propagating and amplifying inflammatory responses to CoV2. Using single cell RNA sequencing, we will assay mixed populations of cells with infected and non-infected (bystander) cells to determine how virus infection imparts inflammatory gene expression and how inflammatory signals from infected cells affect neighboring uninfected cells to amplify the inflammatory response. Sequencing and bioinformatics analyses will be conducted by the Gale lab transcriptomics and computational biology team, and results will be validated by complimentary techniques. From these studies, we expect to reveal the inflammatory cytokine signaling events following SARS-CoV-2 infection. These studies will also provide biomarkers to identify patients susceptible to cytokine storm and will identify gene or gene network/pathway targets to inform strategies to treat COVID-19 disease.

每年，人类都会遇到各种季节性冠状病毒，这些病毒会引起类似于流感的轻微症状 相比之下，导致 COVID-19（2019 年冠状病毒病）的 SARS-CoV-2 则已感染。 截至 2020 年 6 月 4 日，仅在美国就造成了超过 11 万人死亡。由于未知原因，新冠肺炎 - 19 疾病严重程度差异极大，许多感染者出现轻微症状，包括 然而，相当数量的感染者会出现喉咙痛、头痛、咳嗽或根本没有症状。 经历严重疾病，包括急性呼吸窘迫综合征、肾衰竭、心脏骤停和 重要的是，SARS-CoV-2 感染会引发过度活跃的免疫反应。 导致细胞因子风暴，这是促炎分子的大量释放，可能导致致命的组织 损害并可能导致最严重的疾病症状的是触发和调节细胞因子风暴的因素。 尚不清楚，需要研究来揭示 SARS-CoV-2 诱发炎症的过程，以及如何 因此，该提案的首要目标是确定 SARS-CoV-2 感染会传染 COVID19 疾病。 针对 SARS-CoV-2 的炎症反应是如何产生的 为了实现这一目标，我们提出以下建议。 具体目标： 1. 定义 SARS-CoV-2 诱导的炎症细胞因子反应 我们将评估细胞因子。 使用来自肺部和免疫细胞的细胞系，从感染 SARS-CoV-2 的细胞中进行生产 此外，因为重症 COVID-19 患者的细胞因子 IL-1β 和细胞因子水平升高。 因为 IL-1β 在称为细胞焦亡的炎症形式的细胞死亡后释放，我们将评估 然后我们将使用基于 CRISPR 的基因敲除和 SARS-CoV-2 刺激细胞焦亡的能力。 药物抑制剂来绘制观察到的产生和释放所需的分子途径 细胞因子。 2. 确定感染细胞和旁观细胞在传播和传播过程中的炎症转录组 我们将使用单细胞 RNA 测序来放大对 CoV2 的炎症反应。 具有受感染和未受感染（旁观者）细胞的细胞群，以确定病毒感染如何传播 炎症基因表达以及来自受感染细胞的炎症信号如何影响邻近的未感染细胞 细胞放大炎症反应将由测序和生物信息学分析进行。 Gale 实验室转录组学和计算生物学团队，结果将通过免费验证 技术。 通过这些研究，我们期望揭示 SARS-CoV-2 后的炎症细胞因子信号转导事件 这些研究还将提供生物标志物来识别易受细胞因子风暴影响的患者。 识别基因或基因网络/通路目标，为治疗 COVID-19 疾病的策略提供信息。