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，导致新冠肺炎(2019年冠状病毒病)的SARS-CoV-2病毒 截至2020年6月4日，仅在美国就造成超过11万人死亡。出于未知原因，COVID- 疾病的严重程度千差万别，许多感染者出现轻微症状，包括 喉咙痛、头痛和咳嗽，或者根本没有症状。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、肾功能衰竭、心脏骤停和 胃肠道不适。重要的是，SARS-CoV-2感染可以触发过度活跃的免疫反应， 导致细胞因子风暴，这是一种可以导致致命组织的促炎分子的大量释放 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，还需要研究来揭示SARS-CoV-2引起炎症的过程以及如何 SARS-CoV-2感染导致COVID19疾病。因此，这项提案的首要目标是确定 SARS-CoV-2的炎症反应是如何产生的。为了实现这一目标，我们提出了以下建议 具体目标： 1.明确SARS-CoV-2诱导的炎性细胞因子反应。我们将评估细胞因子 利用来自肺的细胞系和居住的免疫细胞从感染SARS-CoV-2的细胞中产生 肺部。此外，由于重症新冠肺炎患者存在细胞因子IL-1β和 由于IL-1β是在一种称为下垂的炎性细胞死亡形式之后释放的，我们将评估 SARS-CoV-2刺激下垂的能力。然后我们将使用基于CRISPR的基因敲除和 药物抑制剂，以绘制所观察到的药物产生和释放所需的分子途径 细胞因子。 2.检测感染细胞和旁观者细胞中的炎性转录组 放大对CoV2的炎症反应。使用单细胞RNA测序，我们将分析混合 用感染和非感染(旁观者)细胞群来确定病毒感染是如何传播的 感染细胞的炎症基因表达和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 盖尔实验室转录学和计算生物学团队，结果将通过免费 技巧。 从这些研究中，我们期望揭示SARS-CoV-2之后的炎性细胞因子信号事件 感染。这些研究还将提供生物标记物来识别易受细胞因子风暴影响的患者，并将 识别基因或基因网络/途径靶点，为治疗新冠肺炎病的策略提供信息。