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β-冠状病毒SARS-CoV-2引起的致命COVID-19疾病表现为 呼吸窘迫与血氧饱和度水平的快速下降，虽然其他 器官缺陷经常在患者和临床病例系列中描述。人类的描述性研究 患者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，并释放炎性细胞因子。 然而，这些假设仍然纯粹是相互关联的，因为因果关系不能严格建立在 人类病人小鼠遗传方法尚未用于测试COVID-19 致病机制为了解决这一知识差距，我们生成了新的小鼠遗传模型 其表达来自小鼠Ace 2基因座的hACE 2，其表达水平足以引起致死性疾病，缺氧， 和肺血管血栓形成，如在人类患者中观察到的。 最初的资助计划是研究这些现象的血管机制， 包括1.2）确定急性ARDS表型或 SARS-CoV 2感染引起的长期肺损伤; 1.3）确定血管细胞感染的贡献 对于急性ARDS表型或SARS-CoV 2感染造成的长期肺损伤;和3）比较 并对比流感与SARS-CoV-2对肺血管和血栓形成的影响。 随后，我们探讨了COVID-19对肺血管内 使用与我们的hACE 2小鼠繁殖的PAR 1-Tango报告基因等位基因进行血栓形成，并观察到 多个器官（肺、肾、肝）血管床中PAR 1-Tango活性异常， SARS-CoV-2感染，这是意料之外的，因为明显的血栓仅存在于肺部。 除了这些新的发现，我们还了解到使用Cre/lox介导的内皮细胞缺失， hACE 2，SARS-CoV-2血管感染不太可能导致我们 观察本补充文件提出了额外的工作，以便：a）揭示的程度和严重性， COVID-19疾病中的血管损伤，使用PAR 1-Tango标本; B）了解实际 利用广泛的多器官系统研究SARS-CoV-2感染中凝血酶产生的机制 免疫组织学检查;和c）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 该补编已收集，但有待深入分析，非常适合扩大联合国的工作， 但在原有的范围内。不需要额外的设备或试剂来追踪 这些研究，而不是需要候选人的时间和精力以及资源 在Kahn实验室已经有了。