权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Genetic Investigation of Covid 19 in Lung Disease

Covid 19 在肺部疾病中的基因研究

基本信息

批准号：
10502908
负责人：
MARK L KAHN
金额：
$ 92.28万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10502908
关键词：
2019-nCoV ACE2 AGTR2 gene Acute Acute Respiratory Distress Syndrome Address Alveolar American Animal Model Autopsy Binding Biological Assay Blood Blood Vessels COVID-19 COVID-19 mortality COVID-19 pandemic COVID-19 pathogenesis COVID-19 severity COVID-19 test Cause of Death Cell Lineage Cell Surface Proteins Cells Cessation of life Chronic Chronic lung disease Consensus Cultured Cells Defect Development Disease Disease Outbreaks Disease model Engineering Epithelial Epithelial Cells Etiology Event Extracorporeal Membrane Oxygenation Foundations Functional disorder Future Gene Expression Genetic Genetic Models Human Hypoxemia Hypoxemic Respiratory Failure Hypoxia Infection Inflammatory Influenza Investigation K-18 conjugate Knock-in Knock-in Mouse Knowledge Long COVID Lung Lung diseases Maps Mechanical ventilation Mediating Modeling Molecular Mus Observational Study Organ Oxygen Pathogenesis Pathogenicity Pathology Patients Phenotype Primary Infection Reporting Respiratory System Respiratory Tract Infections Respiratory distress Role SARS-CoV-2 infection Sampling Secondary to Slice Solid Specificity Testing Thrombosis Tissues Transgenes Transgenic Organisms Vascular Diseases Viral Virus acute infection base betacoronavirus cell type cytokine defined contribution design disease phenotype experience genetic approach human disease human model influenza infection insight knockin animal lung injury mouse genetics mouse model novel novel therapeutic intervention novel therapeutics post SARS-CoV-2 infection prevent protein expression pulmonary vascular cells pulmonary vascular disorder response supplemental oxygen thrombotic vascular contributions

项目摘要

SUMMARY The COVID-19 pandemic due to the SARS-CoV-2 virus has resulted in millions of deaths worldwide. The majority of COVID-19 deaths are caused by lung disease characterized by alveolar filling and severe hypoxemia. Descriptive studies of human patients, animal models and cultured cells have supported numerous pathogenic mechanisms for COVID-19 lung disease, including direct epithelial cell infection, vascular cell infection and thrombosis, and acute respiratory distress syndrome. Despite intense investigation, these hypotheses remain unproven due to a lack of cellular functional evidence for causality. SARS-CoV-2 infection requires viral binding of the human ACE2 (hACE2) cell surface protein, and wild-type virus cannot bind mouse ACE2. Existing hACE2- expressing mice either drive disease through non-endogenous transgenes that do not permit conditional analysis or fail confer severe illness after infection with SARS-CoV-2. Thus, powerful mouse genetic approaches have not yet been harnessed to test COVID-19 pathogenic mechanisms. To address this gap in knowledge we have generated new mouse genetic models that (i) express hACE2 from the mouse Ace2 locus at levels sufficient to confer lethal disease and hypoxia like that observed in human patients, (ii) permit Cre-mediated loss of hACE2 expression to functionally identify cells required to confer COVID-19 disease, and (iii) permit-Cre mediated gain of hACE2 expression to functionally identify cells sufficient for COVID-19 disease. Our preliminary studies identify both epithelial cell infection and vascular disease associated with extensive intravascular thrombosis in the lungs of hACE2 knockin animals. We therefore hypothesize that COVID-19 lung disease arises due to synergistic infection and/or dysfunction of both lung epithelial and lung vascular cells. To test this central hypothesis we will (i) use established Cre-expressing transgenes to test the requirement(s) for epithelial and vascular cell types during COVID-19 lung disease, (ii) use lung slice explants from human and hACE2 knockin mouse lungs to map the cells infected by SARS-CoV-2 virus, and (iii) compare acute and chronic lung responses to infection by the influenza and SARS-CoV-2 viruses to identify the pathogenic mechanisms that underlie the exceptional lethality of COVID-19 lung disease. These studies are expected to yield a functional and integrated understanding of the events that underlie COVID-19 lung disease and provide a solid scientific foundation for the development of novel therapeutic approaches.

概括由 SARS-CoV-2 病毒引起的 COVID-19 大流行已导致全球数百万人死亡。大多数的 COVID-19 死亡是由以肺泡充盈和严重低氧血症为特征的肺部疾病引起的。对人类患者、动物模型和培养细胞的描述性研究支持了许多致病性 COVID-19肺部疾病的机制，包括直接上皮细胞感染、血管细胞感染和血栓形成和急性呼吸窘迫综合征。尽管进行了深入的调查，这些假设仍然存在由于缺乏因果关系的细胞功能证据而未经证实。 SARS-CoV-2 感染需要病毒结合人类ACE2（hACE2）细胞表面蛋白的结合，而野生型病毒不能结合小鼠ACE2。现有的hACE2- 表达小鼠要么通过不允许条件分析的非内源转基因驱动疾病或未能在感染 SARS-CoV-2 后导致严重疾病。因此，强大的小鼠遗传学方法已经尚未用于测试 COVID-19 致病机制。为了解决这一知识差距，我们有产生了新的小鼠遗传模型，该模型（i）从小鼠 Ace2 基因座表达 hACE2，其水平足以导致致命疾病和缺氧，就像在人类患者中观察到的那样，(ii) 允许 Cre 介导的 hACE2 损失表达以功能性识别导致 COVID-19 疾病所需的细胞，以及 (iii) 允许 Cre 介导的增益 hACE2 表达，以在功能上识别足以应对 COVID-19 疾病的细胞。我们的初步研究识别上皮细胞感染和与广泛血管内血栓形成相关的血管疾病 hACE2 敲入动物的肺部。因此，我们假设 COVID-19 肺部疾病的发生是由于肺上皮细胞和肺血管细胞的协同感染和/或功能障碍。为了测试这个中心假设我们将 (i) 使用已建立的表达 Cre 的转基因来测试上皮细胞和 COVID-19 肺部疾病期间的血管细胞类型，(ii) 使用来自人类和 hACE2 敲入的肺切片外植体小鼠肺部绘制被 SARS-CoV-2 病毒感染的细胞图谱，以及 (iii) 比较急性和慢性肺部反应流感和 SARS-CoV-2 病毒感染，以确定其致病机制 COVID-19 肺部疾病具有极高的致死率。这些研究预计将产生功能性和综合性的了解导致 COVID-19 肺部疾病的事件，并为以下方面提供坚实的科学基础：开发新的治疗方法。