NITRIC OXIDE AND SMOOTH MUSCLE CELL PROLIFERATION

我的目标是成为一名学术性的血管外科医生，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，包括我的博士学位。研究表明，一氧化氮可抑制血管平滑肌细胞增殖，这是一种与血管手术临床相关的过程。平滑肌细胞的增殖对于动脉粥样硬化和手术后疾病如再狭窄和静脉移植失败的发展都是至关重要的。在这个建议中，我将使用各种分子方法，包括显性负突变体和反义结构，以及激酶和细胞周期蛋白活性的分析，以确定参与一氧化氮抗增殖作用的细胞内机制。具体目标是：1）检测环核苷酸和环核苷酸依赖性激酶在介导一氧化氮的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表达的影响，（例如fos、myc、myb）和抑制原癌基因表达在一氧化氮抑制生长中的潜在作用，3）探讨一氧化氮诱导细胞周期蛋白依赖性激酶抑制剂p21的机制及其在一氧化氮介导生长停滞中的作用。将在用外源性一氧化氮处理的培养的平滑肌细胞中评价这些细胞内途径的调节作用。在我的赞助商的指导下进行的这些调查，加上这个研究职业奖的课程，研讨会和合作，将大大拓宽我的科学经验，并使我能够通过广泛的基础实验技术来解决血管生物学中的未来问题。这些机制的定义是很重要的，了解病理性平滑肌细胞增殖的生理抑制内皮细胞和潜在的新的治疗策略，增殖性血管疾病。

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.