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

补充氧气常用于治疗肺功能不全的患者， 急性呼吸窘迫综合征（ARDS）是急性肺损伤（ALI）的一种严重形式， 全球数百万。高氧肺损伤被认为是ALI/ARDS的理想模型。的 本申请中提出的研究的中心假设是， NF-E2相关因子（Nrf 2）和/或NADPH醌还原酶基因中的多态性（SNP） NQO 1通过减弱肺、肝功能指标的表达而参与ALI/ARDS的发生 II相抗氧化酶，导致氧介导的活性氧的形成增加 种（ROS），这反过来又导致对ALI/ARDS的易感性增加，以及加重 这些患者的肺损伤。为了实现这些目标，我们提出以下具体建议： 目的：1.为了验证在Nrf 2或NQO 1基因上携带特定SNP的人将被认为是人类的假设， 更容易发展ALI/ARDS比那些谁没有，这些人将显示 比携带野生型基因的人对氧化应激的敏感性增加。气管内抽吸物 将分析来自患有ALI/ARDS的个体或对照的SNP、F2- 异前列烷/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 还将使用来自这些个体的气管吸出物的RNA研究表达谱。2.到 确定缺乏Nrf 2和NQO 1基因的小鼠更易受影响的机制 高氧肺损伤，并测试假设，细胞色素P4501 A（CYP 1A）诱导剂[例如，¿- 萘酮（BNF）]或奥美拉唑（OM）将拯救缺乏功能性Nrf 2或NQO 1基因的小鼠 对抗高氧肺损伤，通过新的机制，需要肝和肺CYP 1A酶。 3.为了确定Nrf 2或NQO 1启动子上已知的SNP调节 人肺细胞或人源化小鼠体内的氧毒性。该目标有两个子目标。（i）.到 测试携带Nrf 2或NQO 1基因的已知SNP的人肺细胞将更多的假设， 易受氧中毒影响。(ii)为了创造转基因人源化小鼠， Nrf 2基因或NQO 1基因或在这些基因上携带已知SNP的那些，并确定 高氧肺损伤中的SNPs组学方法，包括基因组学（微阵列）和蛋白质组学 方法将用于确定Nrf 2或NQO 1变体的分子机制， 会导致肺损伤成功地实现这些目标可以导致创新战略， 新的生物标志物以及新方法的开发（例如，使用PPI，如OM）， 预防/治疗人类的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.