Functional roles of Nrf2 and NQO1 genetic variants in hyperoxic lung injury-ARDS

补充氧气经常用于治疗慢性阻塞性肺疾病患者的肺功能不全 急性呼吸窘迫综合征(ARDS)，这是急性肺损伤(ALI)的一种严重形式，影响 全球数百万人。高氧性肺损伤被认为是ALI/ARDS的合适模型。这个 在本申请中提出的研究的中心假设是特定的单核苷酸 核因子-E2相关因子(NRF2)和/或NADPH苯醌还原酶基因的SNPs (NQO1)通过降低肺和肝功能的表达参与ALI/ARDS的发病。 第二相抗氧化酶，导致氧介导的活性氧的形成增加 物种(ROS)，这反过来导致ALI/ARDS易感性增加，并加剧 这些患者的肺损伤。为了实现这些目标，我们提出了以下具体建议 目的：1.检验以下假设：在Nrf2或NQO1基因上携带特定SNPs的人将是 比那些没有ALI/ARDS的人更容易患上ALI/ARDS，这些人会表现出 比那些携带野生型基因的人对氧化应激更敏感。气管内吸入物 将分析患有ALI/ARDS或对照组的个体是否存在SNPs，F2- 异前列腺素/异呋喃、大体积氧化DNA加合物和蛋白质氧化产物的水平。基因 还将使用这些个体的气管抽吸物中的RNA来研究表达谱。2.至 确定缺乏Nrf2和NQO1基因的小鼠更易受影响的机制 高氧性肺损伤，并验证细胞色素P4501a(CyP1a)诱导剂[例如，？ 或奥美拉唑(OM)将拯救缺乏功能性Nrf2或NQO1基因的小鼠 抗高氧性肺损伤，通过新的机制需要肝脏和肺的CYP1A酶。 3.确定已知的Nrf2或NQO1启动子上的SNPs调控机制 人肺细胞或人源化小鼠体内的氧毒性。这个目标有两个子目标。(I)。至 检验人类肺细胞携带已知的Nrf2或NQO1基因SNPs的假设 易受氧中毒的。(Ii)创造表达正常人的转基因人源化小鼠 Nrf2基因或NQO1基因或那些在这些基因上携带已知SNPs的基因，并确定其作用 SNPs与高氧性肺损伤组学的方法，包括基因组学(微阵列)和蛋白质组学 将使用方法来确定Nrf2或NQO1变体的分子机制 会导致肺损伤。这些目标的成功实现可能会带来创新的战略 开发新的生物标记物以及新的方法(例如，使用PPI，如OM) 预防/治疗人类ALI/ARDS。

Supplemental oxygen is frequently used in the treatment of pulmonary insufficiency in patients with acute respiratory distress syndrome (ARDS), which is a severe form of acute lung injury (ALI), affecting millions worldwide. Hyperoxic lung injury is recognized as an appropriate model for ALI/ARDS. The central hypothesis of the research proposed in this application is that specific single nucleotide polymorphisms (SNPs) in the genes for NF-E2-related factor (Nrf2) and/or NADPH quinone reductase (NQO1) contribute to ALI/ARDS by attenuating the expression of pulmonary and hepatic functional phase II anti-oxidant enzymes, leading to increased formation of oxygen-mediated reactive oxygen species (ROS), which in turn results in increased susceptibility to ALI/ARDS, as well as exacerbated lung damage in these patients. In order to achieve these goals, we propose the following Specific Aims: 1. To test the hypothesis that humans carrying specific SNPs on the Nrf2 or NQO1 genes will be more susceptible to develop ALI/ARDS than those who do not, and these individuals will display increased sensitivity to oxidative stress than those carrying the wild type genes. Endotracheal aspirates from individuals suffering from ALI/ARDS or controls will be analyzed for the presence of SNPs, F2- isoprostanes/isofurans, levels of bulky oxidative DNA adducts, and protein oxidation products. Gene expression profiles will also be studied using RNA from tracheal aspirates of these individuals. 2. To determine the mechanisms by which mice lacking the genes for Nrf2 and NQO1 are more susceptible to hyperoxic lung injury, and test the hypothesis that the cytochrome P4501A (CYP1A) inducer [e.g., ¿- napthoflavone (BNF)] or omeprazole (OM) will rescue the mice lacking functional Nrf2 or NQO1 genes against hyperoxic lung injury, via novel mechanisms entailing hepatic and pulmonary CYP1A enzymes. 3. To determine the mechanisms by which known SNPs on the Nrf2 or NQO1 promoter modulate oxygen toxicity in human lung cells or in humanized mice in vivo. This aim has two sub-aims. (i). To test the hypothesis that human lung cells carrying known SNPs of Nrf2 or NQO1 gene will be more susceptible to oxygen toxicity. (ii) To create transgenic humanized mice expressing the normal human Nrf2 gene or NQO1 gene or those carrying known SNPs on these genes, and determine the role of SNPs in hyperoxic lung injury. Omics' approaches, including genomics (microarrays) and proteomics approaches will be used to determine the molecular mechanisms by which Nrf2 or NQO1 variants contribute to lung injury. Successful accomplishment of the aims could lead to innovative strategies for the development of novel biomarkers as well as new approaches (e.g., use of PPI such as OM) for the prevention/treatment of ALI/ARDS in humans.