Genetic Investigation of Covid 19 in Lung Disease

Covid 19 在肺部疾病中的基因研究

• 批准号: 10768221
• 负责人: MARK L KAHN
• 金额: $ 16.89万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 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/10768221
• 关键词: 2019-nCoV; ACE2; Acute; Acute Respiratory Distress Syndrome; Address; Alleles; Blood; Blood Vessels; Breeding; COVID-19; COVID-19 test; Case Series; Cell Lineage; Cells; Clinical; Coagulation Process; Defect; Disease; Endothelium; Epithelial Cells; Equipment; Etiology; Exhibits; F2R gene; Generations; Genetic; Genetic Models; Genomics; Histologic; Human; Hypoxia; Infection; Inflammatory; Influenza; Investigation; Kidney; Knowledge; Laboratories; Learning; Liver; Lung; Lung diseases; Mediating; Mus; Organ; Oxygen; Pathogenicity; Pathology; Patients; Phenotype; Production; Pulmonary Pathology; Reagent; Reporter; Resources; Respiratory distress; SARS-CoV-2 infection; Severities; Source; Specimen; Surveys; Techniques; Thrombin; Thrombosis; Thrombus; Time; Tissues; Work; betacoronavirus; cytokine; defined contribution; genetic approach; interest; lung injury; mouse genetics; mouse model; nonhuman primate; parent grant; post SARS-CoV-2 infection; thrombotic; transcriptome sequencing; vascular bed; vascular contributions; vascular injury

Project Summary Lethal COVID-19 disease caused by the beta-coronavirus SARS-CoV-2 is manifest as respiratory distress associated with a rapid decline in blood oxygen saturation levels, though other organ defects are often described in patients and clinical case series. Descriptive studies of human patients have proposed numerous pathogenic mechanisms for these findings, including direct epithelial cell infection, vascular cell infection and thrombosis, and released inflammatory cytokines. However, these hypotheses remain purely correlative as causality cannot be rigorously established in human patients. Mouse genetic approaches have not yet been harnessed to test COVID-19 pathogenic mechanisms. To address this gap in knowledge we generated new mouse genetic models that express hACE2 from the mouse Ace2 locus at levels sufficient to confer lethal disease, hypoxia, and pulmonary vascular thrombosis like that observed in human patients. The parent grant initially proposed to work on vascular mechanisms of these phenomenon, including to 1.2) Determine the epithelial cell lineages necessary for either acute ARDS phenotype or long term lung damage by SARS-CoV2 infection; 1.3) Define the contribution of vascular cell infection for either acute ARDS phenotype or long term lung damage by SARS-CoV2 infection; and 3) Compare and contrast the lung vascular and thrombotic effects of influenza versus SARS-CoV-2. Subsequently, we have explored the mechanism of COVID-19 on pulmonary intravascular thrombosis using the PAR1-Tango reporter allele bred with our hACE2 mice, and have observed abnormal PAR1-Tango activity in the vascular beds of multiple organs (lung, kidney, liver) following SARS-CoV-2 infection, which is unexpected since obvious thrombi are only present in the lungs. Besides these new findings, we have also learned using Cre/lox-mediated endothelial deletion of hACE2, that vascular infection by SARS-CoV-2 is not likely responsible for any major phenotypes we observed. This supplement proposes additional work in order to: a) reveal the extent and severity of vascular injury in COVID-19 disease, using PAR1-Tango specimens; b) understand the actual mechanisms of thrombin generation in SARS-CoV-2 infection by employing an extensive multi-organ immunohistological survey; and c) confirm sources of pro-thrombotic or inflammatory factors using RNA sequencing from the tissues and/or regions of interest. As the majority of specimens are already collected, but awaiting in-depth analysis, the supplement is well-suited to extend the work of the parent grant, but within its original scope. Additional equipment or reagents are not needed to pursue these studies, which instead require the time and effort of the candidate together with resources already available in the Kahn laboratory.

项目摘要 由Beta-Coronavirus SARS-COV-2引起的致命性共证疾病表现为 与血氧饱和水平迅速下降有关的呼吸窘迫，尽管其他 器官缺陷通常在患者和临床病例系列中描述。人类的描述性研究 患者为这些发现提出了许多致病机制，包括直接 上皮细胞感染，血管细胞感染和血栓形成，并释放炎症细胞因子。 但是，这些假设仍然纯粹是相关的，因为不能严格确定因果关系 人类患者。尚未利用小鼠遗传方法来测试COVID-19 致病机制。为了解决知识的差距，我们生成了新的鼠标遗传模型 从小鼠ACE2基因座表达HACE2的水平足以赋予致命疾病，缺氧， 和肺血管血栓形成，就像人类患者中所观察到的那样。 父母赠款最初提议从事这些现象的血管机制， 包括1.2）确定急性ARDS表型或 SARS-COV2感染长期肺损伤； 1.3）定义血管细胞感染的贡献 对于SARS-COV2感染，急性ARDS表型或长期肺损伤； 3）比较 并将流感与SARS-COV-2的肺血管和血栓形成作用对比。 随后，我们探索了Covid-19在肺内血管内的机制 使用我们的HACE2小鼠繁殖的Par1-Tango记者等位基因的血栓形成，并观察到 随后的多个器官（肺，肾脏，肝脏）的血管床中的异常PAR1-TANGO活性 SARS-COV-2感染，这是出乎意料的，因为明显的血栓仅存在于肺中。 除了这些新发现，我们还学习了使用CRE/LOX介导的内皮删除 HACE2，SARS-COV-2感染的血管感染不太可能对任何主要表型负责 观察到。这种补充提出了其他工作，以：a）揭示 使用PAR1-TANGO标本，Covid-19疾病中的血管损伤； b）了解实际 通过使用广泛的多器官 免疫组织学调查； c）使用使用 来自感兴趣的组织和/或区域的RNA测序。因为大多数标本已经 收集但正在等待深入分析，该补充剂非常适合扩展 父母赠款，但在其原始范围内。不需要其他设备或试剂来追求 这些研究需要候选人的时间和精力以及资源 已经在Kahn实验室提供。