NITRIC OXIDE AND SMOOTH MUSCLE CELL PROLIFERATION

我的目标是成为一名学术血管外科医生，拥有独立的研究实验室，并在大学环境中教授血管外科。我的长期研究兴趣，包括我的博士研究，一直是一氧化氮抑制血管平滑肌细胞增殖，这是一个与血管外科临床相关的过程。平滑肌细胞的增殖是动脉粥样硬化和手术后疾病，如再狭窄和静脉移植失败的关键。在这项建议中，L将使用多种分子方法，包括显性负性突变和反义结构，以及对激酶和细胞周期蛋白活性的分析，来确定一氧化氮抑制增殖作用的细胞内机制。其具体目的是：1)研究环核苷酸和环核苷酸依赖的激酶在介导一氧化氮抗增殖作用中的作用；2)确定一氧化氮对细胞周期依赖性原癌基因(如fos、myc、myb)表达的影响以及抑制原癌基因表达在一氧化氮抑制生长中的潜在作用；3)确定一氧化氮诱导细胞周期蛋白依赖性激酶抑制因子p21的机制以及p21在介导一氧化氮抑制生长中的重要性。这些细胞内通路的调节作用将在外源性一氧化氮处理的培养的平滑肌细胞中进行评估。在我的赞助商的指导下进行的这些研究，再加上本次研究生涯奖中的课程、研讨会和合作，将极大地拓宽我的科学经验，并使我能够用一系列基本的实验技术来处理未来的血管生物学问题。这些机制的定义对于理解内皮细胞对病理性平滑肌细胞增殖的生理性抑制以及对增殖性血管疾病潜在的新的治疗策略都是重要的。

My goal is to be an academic vascular surgeon who has an independent research laboratory and teaches vascular surgery in a university setting. My long-term research interest, including my Ph.D. studies, has been inhibition of vascular smooth muscle Cell proliferation by nitric oxide, a process clinically relevant to vascular surgery. Proliferation of smooth muscle cells is critical to the development of both atherosclerosis and post- surgical disorders such as restenosis and vein graft failure. In this proposal l will use a variety of molecular approaches, including dominant negative mutants and anti sense constructs, as well as analysis of kinase and cyclin activity, to define the intracellular mechanisms involved in the antiproliferative effect of nitric oxide. The Specific Aims are: 1) To examine the role of cyclic necleotides and cyclic necleotide-dependent kinases in mediating the anti-proliferative effects of nitric oxide, 2) To determine the effect of nitric oxide on expression of the cell cycle- dependent proto-oncogenes (e.g. fos, myc, myb) and the potential role of inhibition of proto-oncogene expression in growth inhibition by nitric oxide, 3) To determine the mechanism of induction of the cyclin- dependent kinase inhibitor p21 by nitric oxide and the importance of p21 in mediating growth arrest by nitric oxide. The effects of modulation of these intracellular pathways will be evaluated in cultured smooth muscle cells treated with exogenous nitric oxide. These investigations under the guidance of my sponsors, coupled with the courses, seminars and collaborations in this Research Career Award, will substantially broaden my scientific experience and allow me to approach future problems in vascular biology with a wide array of fundamental experimental techniques. Definition of these mechanisms is important to understanding both the physiologic inhibition of pathologic smooth muscle cell proliferation by endothelium and potential new therapeutic strategies for proliferative vascular disorders.