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：
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
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/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

项目摘要

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.

项目概要由 β 冠状病毒 SARS-CoV-2 引起的致命性 COVID-19 疾病表现为呼吸窘迫与血氧饱和度水平快速下降有关，尽管其他原因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这些发现的多种致病机制，包括直接的上皮细胞感染、血管细胞感染和血栓形成，并释放炎症细胞因子。然而，这些假设仍然是纯粹相关的，因为因果关系不能严格地建立在人类患者。小鼠遗传学方法尚未用于测试 COVID-19 致病机制。为了解决这一知识空白，我们生成了新的小鼠遗传模型从小鼠 Ace2 基因座表达 hACE2 的水平足以导致致命疾病、缺氧、和肺血管血栓形成，类似于在人类患者中观察到的情况。家长资助最初提议研究这些现象的血管机制，包括 1.2) 确定急性 ARDS 表型或 SARS-CoV2 感染引起的长期肺损伤； 1.3）定义血管细胞感染的贡献对于急性 ARDS 表型或 SARS-CoV2 感染引起的长期肺损伤； 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对肺血管内的作用机制。使用与我们的 hACE2 小鼠饲养的 PAR1-Tango 报告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 PAR1-Tango 活性异常 SARS-CoV-2 感染，这是出乎意料的，因为明显的血栓仅存在于肺部。除了这些新发现之外，我们还了解到使用 Cre/lox 介导的内皮缺失 hACE2，SARS-CoV-2 的血管感染不太可能导致我们发现的任何主要表型观察到。本补充建议进行额外的工作，以便： a) 揭示问题的程度和严重性使用 PAR1-Tango 标本观察 COVID-19 疾病中的血管损伤； b) 了解实际情况通过广泛的多器官研究 SARS-CoV-2 感染中凝血酶产生的机制免疫组织学调查； c) 使用以下方法确认促血栓或炎症因子的来源对感兴趣的组织和/或区域进行 RNA 测序。由于大部分样本已经已收集，但有待深入分析，该补充非常适合扩展该委员会的工作母基金，但在其原始范围内。不需要额外的设备或试剂来进行这些研究需要候选人投入时间和精力以及资源卡恩实验室已经可用。