Redox Regulation of Endothelial Nitric Oxide Synthase and Cardiovascular Diseases

描述（由申请人提供）：这个职业发展奖项的目的是继续发展陈春安博士的学术生涯，首先作为俄亥俄州立大学心肺研究所的博士后研究员，经过额外的博士后培训，并过渡到心血管研究领域的独立研究者，专门研究NOS功能和氧化还原信号。氧自由基生成增加，可降低一氧化氮的生物利用度，被认为是缺血/再灌注损伤的主要发病机制。在内皮细胞中，内皮型一氧化氮合酶（eNOS）是产生维持心血管功能的关键分子的重要酶。越来越多的证据表明，增加的氧化应激通过氧化翻译后修饰改变了几种酶的功能，如s -谷胱甘肽基化、硝化或亚硝基化，所有这些都与信号转导有关。在指导阶段，Aim1和Aim2将解决几个问题：Aim1是确定eNOS s -谷胱甘肽化和硫醇氧化的详细机制，因为它与心血管疾病有关。目的2是确定Grx1在去谷胱甘肽化过程、氧化还原调控和NOS功能中的作用。从受指导阶段训练中获得的结果将在受指导阶段和独立阶段之间提供一座桥梁。在独立阶段，陈博士的研究将集中在离体小鼠模型上，研究eNOS s -谷胱甘肽化对血管功能的影响，以及Grx1在缺血/再灌注损伤中的作用。目的3：研究eNOS Cys突变体在血管功能障碍期间对血管功能和抗氧化修饰的影响。Aim4是在小鼠离体心脏模型中确定Grx1在缺血/再灌注损伤中的作用，特别是在调节蛋白去谷胱甘肽化和eNOS功能方面的作用。通过了解氧化还原调节酶（Grx）与NO生成之间的关系，这将为理解缺血/再灌注和氧化应激期间心血管功能改变的机制提供关键的一步。成功获得K99/R00奖项将促进陈春安博士的职业研究发展，并使陈春安博士成为心血管研究领域的独立研究者。

DESCRIPTION (provided by applicant): The objective of this career development award is to continue to develop the academic career of Dr. Chun-An Chen, first as a postdoctoral researcher at The Ohio State University Heart and Lung Research Institute, with additional postdoctoral training and transition to an independent investigator in the field of cardiovascular research specializing in NOS function and redox signaling. Increased oxygen free radical generation, which can reduce the bioavailability of nitric oxide, is believed to be the primary pathogenesis of ischemia/reperfusion injuries. In the endothelium, endothelial nitric oxide synthase (eNOS) is the important enzyme that produces this critical molecule maintaining the cardiovascular function. Growing evidence suggests that increased oxidative stress alters the function of several enzymes through oxidative post-translational modifications, such as S-glutathionylation, nitration, or nitrosylation, all of which have been implicated in signal transduction. During the mentored phase, several questions will be addressed in Aim1 and Aim2: Aim1 is to determine the detailed mechanism of eNOS S-glutathionylation and thiol oxidation, as it pertains to cardiovascular diseases. Aim2 is to determine the role of Grx1 in the deglutathionylation process, redox regulation, and NOS function. The results gained from the mentored phase training will provide a bridge between mentored phase and independent phase. During the independent phase, Dr. Chen's research will focus on ex vivo mouse models regarding the effect of eNOS S-glutathionylation on vascular function, and the role of Grx1 on ischemia/reperfusion injury. Several mechanistic questions will be addressed with the following aims: Aim3 is to study the effect of eNOS Cys mutants on vascular function and resistance to oxidative modification during vascular dysfunction. Aim4 is to identify the role of Grx1 in ischemia/reperfusion injury, especially in the regulation of protein deglutathionylation and eNOS function, in mouse ex vivo heart models. By understanding the relationship between a redox regulatory enzyme (Grx) and NO production, this will provide a critical step toward understanding the mechanisms involved in the alteration of cardiovascular function during ischemia/reperfusion and oxidative stress. Successfully securing this K99/R00 award will foster Dr. Chun-An Chen's career research development and enable Dr. Chun-An Chen to become an independent investigator in the field of cardiovascular research. PUBLIC HEALTH RELEVANCE: The objective of this proposed research is to understand the mechanism of reactive oxygen species contributing to coronary heart diseases. Coronary heart disease (CHD) remains the primary killer in the United States, and even though the mortality rate has declined, with the aging population and increased risk factors, the incidence of CHD will likely increase for the next decade. The ultimate goal of this proposed research is to develop new therapeutic strategies against these deadly diseases.