Alcohol and the alveolar epithelial barrier

酒精和肺泡上皮屏障

• 批准号: 10362802
• 负责人: MICHAEL H. KOVAL
• 金额: $ 12.41万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10362802
• 关键词: Active Biological Transport; Acute; Acute Lung Injury; Address; Air; Alcohols; Alveolar; Alveolar Macrophages; Antioxidants; Area; Biology; Cell physiology; Clinical Research; Clinical Trials; Collaborations; Complex; Discipline; Edema; Environment; Epithelial; Equilibrium; Experimental Models; Faculty; Floods; Fluid Balance; Foundations; Funding; Genes; Genetic Transcription; Glutathione; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; HIV; HIV-1; Human; Impairment; Individual; Inflammatory; Injury; Intervention; Investigation; Left; Link; Liquid substance; Lung; Mediating; Morbidity - disease rate; Mothers; Oxidative Stress; Pathway interactions; Permeability; Phase; Phenotype; Pneumonia; Postdoctoral Fellow; Pregnancy; Premature Infant; Proteins; Pulmonary Edema; Regulation; Research Personnel; Respiratory Failure; Response Elements; Role; S-Adenosylmethionine; Scientist; Sepsis; Signal Transduction; Sodium; Stress; Sulforaphane; System; Testing; Therapeutic Intervention; Tight Junctions; Training; Training Programs; Transgenic Animals; Translating; Trauma; Water; Zinc; Zinc deficiency; activating transcription factor; acute stress; alcohol effect; alcohol research; alveolar epithelium; chronic alcohol ingestion; design; equilibration disorder; high risk; immune function; in vivo; in vivo Model; lung injury; macrophage; member; mortality; new therapeutic target; novel; novel therapeutics; oxidant stress; pre-clinical; pre-doctoral; problem drinker; pulmonary function; response; therapeutic development; transcription factor; Active Biological Transport; Acute; Acute Lung Injury; Address; Air; Alcohols; Alveolar; Alveolar Macrophages; Antioxidants; Area; Biology; Cell physiology; Clinical Research; Clinical Trials; Collaborations; Complex; Discipline; Edema; Environment; Epithelial; Equilibrium; Experimental Models; Faculty; Floods; Fluid Balance; Foundations; Funding; Genes; Genetic Transcription; Glutathione; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; HIV; HIV-1; Human; Impairment; Individual; Inflammatory; Injury; Intervention; Investigation; Left; Link; Liquid substance; Lung; Mediating; Morbidity - disease rate; Mothers; Oxidative Stress; Pathway interactions; Permeability; Phase; Phenotype; Pneumonia; Postdoctoral Fellow; Pregnancy; Premature Infant; Proteins; Pulmonary Edema; Regulation; Research Personnel; Respiratory Failure; Response Elements; Role; S-Adenosylmethionine; Scientist; Sepsis; Signal Transduction; Sodium; Stress; Sulforaphane; System; Testing; Therapeutic Intervention; Tight Junctions; Training; Training Programs; Transgenic Animals; Translating; Trauma; Water; Zinc; Zinc deficiency; activating transcription factor; acute stress; alcohol effect; alcohol research; alveolar epithelium; chronic alcohol ingestion; design; equilibration disorder; high risk; immune function; in vivo; in vivo Model; lung injury; macrophage; member; mortality; new therapeutic target; novel; novel therapeutics; oxidant stress; pre-clinical; pre-doctoral; problem drinker; pulmonary function; response; therapeutic development; transcription factor

Project Summary The COVID-19 pandemic represents the most significant public health crisis to occur in generations. A major cause of death due to COVID-19 is acute respiratory distress syndrome (ARDS). Whether chronic alcohol use disorder exacerbates the severity of COVID-19 is not known, but is highly likely given the known impact of alcohol on several organ systems including priming the lung epithelial barrier for increased paracellular leak. In addition, it is well established that chronic alcohol consumption exacerbates the severity of lung injury when combined with an additional insult, a so-called “second hit”. Of note, alcoholic lung syndrome increases the incidence of ARDS by 3-4 fold when compared with non-alcoholic ICU patients. We have identified properties of SARS-CoV-2 that predispose it towards a milder phase of infection, related to the virus E protein. The original SARS coronavirus, SARS-CoV-1, has an E protein that specifically targets tight junction protein PDZ binding motifs and thus readily disrupts the epithelial barrier of infected epithelia. By contrast, the COVID-19 virus SARS-CoV-2 has E protein mutations that inhibit its ability to bind to PDZ binding motifs which can help preserve infected epithelial cell barrier function. However, SARS-CoV-2 in the context of alcoholic lung syndrome is more likely to result in a more severe outcome, since the epithelial barrier is already impaired as a result of chronic alcohol abuse. Primary human bronchiolar epithelial cells derived from alcoholic and non- alcoholic subjects will be infected with SARS-CoV-2 in vitro and we will measure several outcome variables related to severity of infection. We also have preliminary data showing that cells from alcoholics retain signature differences in gene expression and function relative to non-alcoholic cells, leading to decreased barrier function and altered cell morphology. This supports a hypothesis that chronic alcohol exposure induces persistent, epigenetic changes to the airway epithelial cell genome. In this supplement proposal, we will determine the impact of SARS-CoV-2 on the epigenome of host cells from alcoholics and non-alcoholics, focusing on DNA methylation. Of particular interest will be to examine the impact of DNA methylation induced by alcohol and/or SARS-CoV-2 infection on the expression and downstream targets of the transcription factors Nrf2 and PU.1, known to be impaired by chronic alcohol exposure and that play key roles in regulating epithelial antiviral responses. We will also identify novel host genome loci showing patterns of differential methylation that correlate with disease severity. We anticipate that identifying differentially methylated loci impacted by chronic alcohol exposure and SARS-CoV-2 infection will lead to drug targets with the potential to reduce the severity of COVID-19 in alcoholic patients.

项目摘要 COVID-19大流行代表了几代人发生的最重大的公共卫生危机。专业 COVID-19引起的死亡原因是急性呼吸窘迫综合征（ARDS）。是否使用慢性酒精 疾病加剧了COVID-19的严重程度尚不清楚，但鉴于已知的影响很可能 在几种器官系统上的酒精，包括启动肺上皮屏障，以增加细胞细胞泄漏。在 此外，还可以很好地确定，长期饮酒会加剧肺部损伤的严重程度 再加上另外的侮辱，即所谓的“第二击”。值得注意的是，酒精性肺综合征增加 与非酒精性ICU患者相比，ARDS的发病率增加了3-4倍。我们已经确定了属性 SARS-COV-2的易感性倾向于与病毒蛋白有关的米勒感染阶段。 原始SARS冠状病毒SARS-COV-1具有E蛋白，该蛋白专门针对紧密连接蛋白PDZ 结合基序，因此很容易破坏感染上皮的上皮屏障。相比之下，Covid-19 病毒SARS-COV-2具有E蛋白突变，可抑制其与PDZ结合基序结合的能力，这可以帮助 保存感染的上皮细胞屏障功能。但是，在酒精肺的背景下SARS-COV-2 综合征更有可能导致更严重的结果，因为上皮屏障已经受到损害 慢性酒精滥用的结果。原发性人支气管上皮细胞，源自酒精和非酒精 酒精受试者将在体外感染SARS-COV-2，我们将测量几个结果变量 与感染的严重程度有关。我们还拥有初步数据，表明酗酒者的细胞保留 基因表达和功能相对于非酒精细胞的特征差异，导致了改善 屏障功能和细胞形态改变。这支持一个假设，即慢性酒精暴露会诱发 气道上皮细胞基因组的持续性表观遗传变化。在此补充建议中，我们将 确定SARS-COV-2对酗酒和非酒精药物的宿主细胞表观基因组的影响， 专注于DNA甲基化。特别有趣的是检查DNA甲基化诱导的影响 通过酒精和/或SARS-COV-2感染转录因子的表达和下游靶标 NRF2和PU.1，已知会受到慢性酒精暴露的损害，并且在调节中起关键作用 上皮抗病毒反应。我们还将识别出新的宿主基因组基因座，显示差异的模式 与疾病严重程度相关的甲基化。我们预计确定不同的甲基化基因座 受慢性酒精暴露和SARS-COV-2感染的影响将导致药物靶标，并有可能 在酒精患者中降低了COVID-19的严重程度。