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 普通感冒相比之下，导致COVID-19（2019年冠状病毒病）的SARS-CoV-2， 截至2020年6月4日，仅在美国就造成超过11万人死亡。由于未知的原因，COVID- 19疾病的严重程度变化很大，许多感染者出现轻微症状，包括 喉咙痛，头痛，咳嗽或没有任何症状。然而，相当数量的感染者 患有严重疾病，包括急性呼吸窘迫综合征、肾衰竭、心脏骤停，以及 胃肠道不适重要的是，SARS-CoV-2感染可引发过度活跃的免疫反应， 导致细胞因子风暴，这是一种大量释放的促炎分子，可导致致命的组织 损害，并可能导致最严重的疾病症状。触发和调节细胞因子风暴的是 尚不清楚，需要研究来揭示SARS-CoV-2诱导炎症的过程，以及如何 SARS-CoV-2感染导致COVID 19疾病。因此，本提案的总体目标是确定 如何产生对SARS-CoV-2的炎症反应。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 使用来自肺和免疫细胞的细胞系从感染SARS-CoV-2的细胞中生产， 肺此外，由于严重的COVID-19患者存在细胞因子IL-1β水平升高， 由于IL-1β是在细胞死亡的炎症形式（称为细胞焦亡）后释放的，我们将评估IL-1β在细胞凋亡中的作用。 SARS-CoV-2刺激细胞凋亡的能力。然后，我们将使用基于CRISPR的基因敲除， 药理学抑制剂，以绘制所观察到的药物的产生和释放所需的分子途径。 细胞因子 2.确定增殖和增殖过程中感染细胞和旁观者细胞的炎性转录组， 增强对CoV 2的炎症反应。使用单细胞RNA测序，我们将检测混合 感染和未感染（旁观者）细胞的细胞群，以确定病毒感染如何赋予 炎症基因表达以及来自感染细胞的炎症信号如何影响邻近未感染细胞 细胞来放大炎症反应。测序和生物信息学分析将由 Gale实验室转录组学和计算生物学团队，结果将由免费验证 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传导事件 感染这些研究还将提供生物标志物，以识别易受细胞因子风暴影响的患者， 确定基因或基因网络/通路靶标，为治疗COVID-19疾病提供信息。