Mechanisms of exacerbation of COVID-19 pathogenesis in mice expressing human ACE2 by polycyclic aromatic hydrocarbons (PAHs), and its protection by inhibition of soluble epoxide hydrolase (sEH)

项目摘要 严重冠状病毒病(新冠肺炎)是由一种新型的贝塔冠状病毒引起的，现在命名为 SARS-CoV-2。新冠肺炎的特征是未解决的系统性炎症相关的 伴随着危及生命的细胞因子风暴综合征，导致多器官衰竭功能障碍 一些病人。最近的研究表明，吸烟和其他环境因素 污染物会加剧新冠肺炎感染者的呼吸道疾病，但其机制 导致肺部疾病加重的原因尚不清楚。慢性阻塞性肺疾病肺损伤模型的建立 环境化学品(如吸烟)包括使用多环芳烃 碳氢化合物(PAH)，特别是苯并[a]芘(BP)，存在于香烟烟雾中， 炭化牛排、柴油尾气等。在这些型号中，额外的高氧暴露会导致 ARDS样症状加重。目前的数据表明，使用一种可溶的环氧化物 水解酶抑制剂(SEHI)通过阻止水合作用来保护与ARDS相关的肺损伤 抗炎二十烷酸[例如，环氧二十碳三烯酸(EETs)]。中环 在这个应用中提出的假设是BP会加重肺 SARS-CoV-2感染期间的损伤/炎症，以及随后的高氧暴露， 而用sEHI治疗这些小鼠将提供对肺损伤的保护。 将使用单细胞RNA-Seq和FACS方法进行基因表达谱分析 确定BP/SARS冠状病毒介导的肺损伤和炎症的分子途径 2/高氧血症。我们提出了以下具体目标：1.检验转基因 在感染SARS-CoV-2之前接受BP治疗的K18-hACE2小鼠将获得更多 比那些在感染前接受模拟治疗的人更容易受到肺损伤。我们还将测试 假设使用sEHI TPPU治疗将对肺损伤/ARDS具有保护作用 BP/SARS-COV-2/暴露小鼠。使用单细胞rna-seq的基因表达谱将是 特异性肺细胞在BP/SARS-CoV-2介导的肺损伤中的作用 并由sEHI进行保护。2.检验BP/SARS-CoV-2暴露治疗的假说 与正常对照组相比，高氧组小鼠肺损伤进一步加重. 室内空气，如果这些小鼠暴露在sEHI治疗中，它们将表现出较轻的伤害 在高氧期。拟议中的研究将揭开肺的分子机制 SARS-CoV-2/高氧介导的损伤及其环境多环芳烃的增强作用 此外，如果我们的sEHI研究旨在保护小鼠免受新冠肺炎的影响，那么它将是 朝着未来使用SEHS治疗人类柯萨奇病毒019的临床试验迈出了一大步。

Project Summary Severe coronavirus disease (COVID-19) is caused by a novel Beta-coronavirus, now named SARS-CoV-2. COVID-19 is characterized by unresolved systemic hyperinflammation associated with a life-threatening “cytokine storm syndrome”, leading to multi-organ failure dysfunction in some patients. Recent studies have shown that cigarette smoking and other environmental pollutants exacerbate respiratory illness in COVID-19 infected individuals, but the mechanisms responsible for the potentiation of lung disease is not known. Models of lung damage due to environmental chemicals (e.g., cigarette smoking) include the use of polycyclic aromatic hydrocarbons (PAH), especially benzo[a]pyrene (BP), which are present in cigarette smoke, charbroiled steaks, diesel exhausts etc. In these models, additional hyperoxic exposure leads to exacerbation of ARDS-like symptoms. Current data suggests that using a soluble epoxide hydrolase inhibitor (sEHI) protects against lung injury related to ARDS, as they prevent hydration of anti-inflammatory eicosanoids [e.g., epoxy eicasotrienoic acids (EETs). The central hypothesis proposed in this application is that BP would exacerbate lung injury/inflammation during SARS-CoV-2 infection, and subsequent hyperoxia exposure, and that treatment of these mice with sEHI would confer protection against lung injury. Gene expression profiling using single cell RNA-Seq and FACS approaches will be done to determine the molecular pathways of lung injury and inflammation mediated by BP/SARS-CoV- 2/hyperoxia. We propose the following specific aims: 1. To test the hypothesis that transgenic K18-hACE2 mice that are treated with BP prior to infection with SARS-CoV-2 will be more susceptible to lung injury than those that are mock treated prior to infection. We will also test the hypothesis that treatment with the sEHI TPPU will confer protection against lung injury/ARDS in the BP/SARS-COV-2/-exposed mice. Gene expression profiling using single cell RNA-seq will be performed to determine the role of specific lung cells in lung injury mediated by BP/SARS-CoV-2 and its protection by sEHI. 2. To test the hypothesis that exposure of BP/SARS-CoV-2 treated mice to hyperoxia will lead to further exacerbation of lung injury compared to those maintained in room air, and that these mice will display lesser injury if they were exposed to sEHI treatment during the hyperoxia phase. The proposed studies will unravel molecular mechanisms of lung injury mediated by SARS-CoV-2/hyperoxia, and its potentiation by environmental PAHs. Furthermore, if our sEHI studies aimed to protect mice against COVID-19 pathogenesis, it will be a big step towards future clinical trials on the use of sEHs for treatment of COVID019 in humans.