Redox Regulation of Endothelial Nitric Oxide Synthase and Cardiovascular Diseases

描述(申请人提供)：该职业发展奖的目标是继续发展陈春安博士的学术生涯，首先是作为俄亥俄州立大学心肺研究所的博士后研究员，进行额外的博士后培训，并过渡到心血管研究领域的独立研究员，专门从事一氧化氮合酶功能和氧化还原信号的研究。氧自由基的产生增加，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)是产生这一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，氧化应激增加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的详细机制-谷胱甘肽和硫醇氧化，因为它与心血管疾病有关。AIM2的目的是确定Grx1在谷胱甘肽化过程、氧化还原调节和NOS功能中的作用。指导阶段培训的结果将在指导阶段和独立阶段之间架起一座桥梁。在独立阶段，陈博士的研究将集中在体外小鼠模型上，了解eNOS S谷胱甘肽基化对血管功能的影响，以及Grx1在缺血/再灌注损伤中的作用。AIM3是为了研究eNOS Cys突变体在血管功能障碍时对血管功能和抗氧化性的影响。目的探讨Grx1在小鼠心脏缺血再灌注损伤中的作用，特别是在蛋白谷胱甘肽和eNOS功能调节中的作用。通过了解氧化还原调节酶(GRX)和NO产生之间的关系，这将为理解缺血/再灌注和氧化应激过程中心血管功能变化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员。 公共卫生相关性：这项拟议研究的目的是了解活性氧类对冠心病的作用机制。冠心病(CHD)仍然是美国的主要杀手，尽管死亡率已经下降，但随着人口老龄化和危险因素的增加，CHD的发病率在未来十年可能会增加。这项拟议研究的最终目标是开发针对这些致命疾病的新治疗策略。

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.