The NAD(P)H Oxidase in Airways Remodeling and Reactivity

NAD(P)H 氧化酶在气道重塑和反应中的作用

• 批准号: 6631428
• 负责人: John R Hoidal
• 金额: $ 39.52万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6631428
• 关键词: NAD(P)H dehydrogenase; asthma; bronchomotion; cell growth regulation; cell proliferation; clinical research; enzyme mechanism; free radical oxygen; gene targeting; genetically modified animals; growth factor; human tissue; laboratory mouse; muscle contraction; nuclear factor kappa beta; respiratory muscles; smooth muscle; tissue /cell culture

DESCRIPTION (provided by applicant): OBJECTIVE: The goals of this project are to identify the roles of endogenous NAD(P)H oxidases in the proliferative responses and enhanced contractility leading to airways smooth muscle (AWSM) remodeling and hyperreactivity in asthma. HYPOTHESIS: The inclusive hypothesis to be tested is that airway smooth muscle NAD(P)H oxidases are highly regulated ROS producing enzymes that play distinct roles in initiating AWSM proliferation and contraction. SPECIFIC AlMS: The first specific aim will characterize the NAD(P)H oxidase(s) of airway smooth muscle (AWSM) cells and determine its contribution to the generation of ROS. This aim will address the hypothesis that NAD(P)H Oxidase 4 (Nox4) bound to membranes in the endoplasmic reticulum is the catalytic subunit of the NAD(P)H oxidase in proliferating AWSM. The second specific aim will define the role of NAD(P)H oxidase(s) in AWSM proliferation. This aim will test the hypotheses that specific growth factors induce AWSM proliferation via activation of the Nox4 oxidase with resultant transactivation of nuclear factor-kappa B (NF-kB). The third specific aim will define the role of NAD(P)H oxidase(s) in AWSM contractile function. This aim will test the hypothesis that ROS generated by an NAD(P)H oxidase induce AWSM contraction, but that components of the oxidase in the contractile phenotype are distinct from those of the proliferative phenotype. RESEARCH PLAN: For the first aim, our strategy is to define and fully characterize the structure of the components responsible for AWSM NAD(P)H oxidase activity and pinpoint their subcellular location and interaction. We will also characterize the activity, the redox midpoint potential of AWSM NAD(P)H oxidase and the role of individual components in the generation of ROS. For the second aim we will utilize purified cells with deficiencies of NAD(P)H oxidase components to directly establish the importance of the oxidase in AWSM proliferation. Emphasis will be placed on establishing the role of the oxidase in transactivation of NF-kB, a factor essential for smooth muscle proliferation. For the third aim we will again utilize purified cells and animals with genetic deficiences of NAD(P)H oxidase components to directly establish the importance and characteristics of the oxidase that regulates airways contractile function. Emphasis will be placed upon investigating whether there is a phenotype switch in the oxidase components. SIGNIFICANCE: The work will provide a better understanding of the importance of AWSM NAD(P)H oxidase in mediating airways smooth muscle remodeling in asthma and contractility in the asthmatic state, with broad importance in the role of ROS in respiratory physiology and disease.

描述（申请人提供）：目的：该项目的目标是 确定内源性NAD（P）H氧化酶在增殖中的作用 响应和增强的收缩性，导致气道平滑肌（AWSM） 哮喘的重塑和过度反应性。假设：包容性假设 要测试的是气道平滑肌NAD（P）H氧化酶是高度调节的 ROS产生的酶在发起AWSM增殖中起着独特的作用 和收缩。特定施舍：第一个特定目标将表征 气道平滑肌（AWSM）细胞的NAD（P）H氧化酶，并确定其 对ROS产生的贡献。这个目标将解决该假设 NAD（P）H氧化酶4（NOX4）与内质网中的膜结合 是增殖AWSM中NAD（P）H氧化酶的催化亚基。这 第二个特定目标将定义NAD（P）H氧化酶在AWSM中的作用 增殖。这个目标将检验特定增长因素的假设 通过激活NOX4氧化酶诱导AWSM增殖 核因子-kappa B（NF-KB）的反式激活。第三个特定目标将 定义NAD（P）H氧化酶在AWSM收缩功能中的作用。这个目标 将检验以下假设，即由NAD（P）H氧化酶产生的ROS诱导AWSM 收缩，但是收缩表型中氧化酶的成分 与增殖表型不同。研究计划： 第一个目的，我们的策略是定义和充分表征 负责AWSM NAD（P）H氧化酶活性的组件并确定其 亚细胞位置和相互作用。我们还将表征活动， AWSM NAD（P）H氧化酶的氧化还原中点潜力和个体的作用 ROS代的组成部分。对于第二个目标，我们将利用 纯化的细胞含有NAD（P）H氧化酶成分的缺乏直接 确定氧化酶在AWSM增殖中的重要性。重点将是 建立氧化酶在NF-kb的反式激活中的作用 平滑肌增殖至关重要的因素。对于第三个目标，我们将 再次利用纯化的细胞和动物，具有NAD（P）H的遗传缺陷 氧化酶成分直接确定 调节气道收缩功能的氧化酶。重点将是 研究是否研究氧化酶中是否有表型开关 成分。意义：这项工作将为您提供更好的理解 AWSM NAD（P）H氧化酶在介导气道平滑肌中的重要性 哮喘状态的哮喘和收缩性重塑 ROS在呼吸生理和疾病中的作用重要性。