Neutrophil Heterogeneity and Immunopathogenesis of COVID-19 ARDS

COVID-19 ARDS 的中性粒细胞异质性和免疫发病机制

• 批准号: 10560925
• 负责人: Jiapeng Huang
• 金额: $ 57.41万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10560925
• 关键词: 2019-nCoV; Acute; Acute Lung Injury; Acute Respiratory Distress Syndrome; Age; Antibodies; Bilateral; Blood; Blood Coagulation Disorders; Blood Platelets; Bronchoalveolar Lavage Fluid; COVID-19; COVID-19 patient; COVID-19 vaccine; COVID-19/ARDS; Cells; Chemotaxis; Circulation; Clinical; Collaborations; Cytometry; Cytoplasmic Granules; Data; Development; Disease; Disease Progression; Economics; Exocytosis; FCGR3B gene; Fibrin fragment D; Functional disorder; Gene Expression; Hemorrhage; Heterogeneity; Hospitalization; Host Defense; Immune; In Vitro; Individual; Inflammation Mediators; Inflammatory; Inflammatory Response; Intensive Care Units; Interleukin-6; Life; Lung; Lymphocyte; Measures; Myeloid-derived suppressor cells; Neutrophil Infiltration; Outcome; Patient Transfer; Patients; Pattern; Personal Satisfaction; Phagocytosis; Plasma; Platelet Activation; Pneumonia; Population; Proteome; Proteomics; Psyche structure; Pulmonary Inflammation; Reactive Oxygen Species; Reporting; Respiratory Burst; Role; Severity of illness; Surface; System; T-Lymphocyte; TNF gene; Therapeutic Intervention; Variant; Viral; antimicrobial; chemokine; cytokine; cytokine release syndrome; density; effective therapy; extracellular; immune activation; immunothrombosis; in vivo; inhibitor; insight; lung injury; mouse model; multiple omics; neutrophil; novel; novel marker; novel therapeutic intervention; pandemic disease; pathogenic virus; pharmacologic; post SARS-CoV-2 infection; prospective; response; severe COVID-19; sex; systemic inflammatory response; therapeutic target; therapeutically effective; tissue injury; transcriptome sequencing; transcriptomic profiling; transcriptomics

Project Summary Coronavirus disease 2019 (COVID-19) is a potentially life-threatening disease caused by the novel viral pathogen, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Of the 10%-20% of COVID-19 patients hospitalized, approximately 1/3 develop acute respiratory distress syndrome (ARDS). COVID-19- induced ARDS results from a combination of virally induced lung injury and the rapid influx of immune cells that release inflammatory mediators leading to a hyper-activated state known as cytokine storm. Understanding the pathophysiology of COVID-19 ARDS is critical to finding effective therapeutic interventions. Accumulating evidence indicates critical roles for neutrophils in both ARDS and immunothrombosis in COVID-19. Multi-omics studies identified dramatic neutrophil heterogeneity in COVID-19, including emergence of a large number of low density neutrophils (LDN). We reported a novel population of LDN present in both the circulation and lungs of severe COVID-19 patients which expresses intermediate levels of CD16 (CD16Int LDN). CD16Int LDN spontaneously form neutrophil extracellular traps (NETs), activate platelets in vivo and in vitro, and have heightened degranulation. Our preliminary data show that LDN subsets from severe COVID-19 patients are functionally distinct from mature, normal density neutrophils (NDN). Based on these findings, we postulate that differential functional responses of heterogeneous neutrophil populations collaborate to induce systemic and pulmonary inflammation leading to ARDS in COVID-19 patients. Three specific Aims are proposed to further dissect the underlying mechanisms. Aim 1 will comprehensively characterize neutrophil subsets from severe/critical COVID-19 patients using proteomics and transcriptomics approaches. The information gained from those studies will be used to refine our CyTOF antibody panel and prospectively measure dynamic changes of neutrophil subsets over the disease course in COVID-19 patients. Aim 2 will determine neutrophil subset functional changes during disease progression and their contributions to dysregulated inflammatory response and coagulopathy in severe/critical COVID-19 patients. Neutrophil degranulation, NET formation, phagocytosis, chemotaxis, T cell suppressive activity, and cytokine/chemokine release will be examined. We will also determine if LDN subsets and NDN or their derived products promote coagulopathy in COVID-19 patients. Aim 3 will determine the relative contributions of neutrophil host defense systems to development of acute lung injury. We will also use a hACE2 Tg mouse model to determine the roles of neutrophil respiratory burst, granule exocytosis, and NET formation to SARS-CoV-2-induced acute lung inflammation. Successful completion of this proposal will provide novel insights into COVID-19-induced ARDS pathophysiology by defining the functional roles of different neutrophil subsets and by establishing which host defense systems of neutrophils as a therapeutic target for inhibiting inflammatory lung injury and immunothrombosis in COVID-19.

项目摘要 冠状病毒疾病2019（Covid-19）是一种潜在的威胁生命的疾病，由新型病毒引起 病原体，严重的急性呼吸道综合征冠状病毒2（SARS-COV-2）。在10％的10％-20％ 住院的患者，大约1/3发展急性呼吸窘迫综合征（ARDS）。新冠肺炎- 诱导的ARDS是由病毒诱导的肺损伤和免疫细胞快速流入的结合而引起的。 释放炎症介质导致过度激活状态称为细胞因子风暴。了解 COVID-19 ARDS的病理生理学对于寻找有效的治疗干预措施至关重要。累积 证据表明中性粒细胞在Covid-19中的ARD和免疫骨栓塞中的关键作用。多词 研究确定了Covid-19的戏剧性嗜中性粒细胞异质性，包括大量出现 低密度中性粒细胞（LDN）。我们报道了循环和肺中存在的新型LDN人群 表达中等水平CD16（CD16INT LDN）的严重共vid-19患者。 CD16INT LDN 自发形成中性粒细胞外陷阱（网），在体内和体外激活血小板，并具有 脱粒增加。我们的初步数据表明，来自严重的Covid-19患者的LDN子集是 在功能上与成熟的正常密度中性粒细胞（NDN）不同。根据这些发现，我们假设 异质性嗜中性粒细胞种群的差异功能反应合作以诱导 全身性和肺部炎症导致COVID-19患者的ARDS。三个具体目标是 提议进一步剖析潜在机制。 AIM 1将全面表征中性粒细胞 使用蛋白质组学和转录组学方法的严重/关键共vid-19患者的子集。这 从这些研究中获得的信息将用于完善我们的细胞抗体面板和前瞻性 测量中性粒细胞亚群的动态变化在COVID-19患者的疾病病程中。 AIM 2意志 确定疾病进展过程中嗜中性粒细胞子集功能变化及其对 严重/关键的COVID患者的炎症反应和凝血病失调。中性粒细胞 脱粒，净形成，吞噬作用，趋化性，T细胞抑制活性和细胞因子/趋化因子 将检查发布。我们还将确定LDN子集和NDN或其衍生产品是否促进 Covid-19患者的凝血病。 AIM 3将确定中性粒细胞宿主防御的相对贡献 发展急性肺损伤的系统。我们还将使用HACE2 TG鼠标模型来确定 中性粒细胞呼吸道爆发的作用，颗粒胞吐作用和净形成SARS-COV-2诱导的急性 肺部炎症。该提案的成功完成将提供有关Covid-19引起的新见解 通过定义不同嗜中性粒细胞亚群的功能作用并确定病理生理学 哪些寄养中性粒细胞的防御系统是抑制炎症性肺损伤和 COVID-19中的免疫骨栓塞。