Nitric Oxide Regulation of Platelet Granule Exocytosis

描述(由申请人提供)： 我正在申请导师临床科学家职业奖(K08)，以获得导师基础研究培训。我希望研究一氧化氮对血小板颗粒胞吐的调节，因为它与血栓形成有关。通过这个奖项，我寻求成为一名能够应用我的知识和培训的研究人员，通过1)一个有指导的基于实验室的组件来获得(A)胞吐研究和(B)血栓形成研究的知识；2)一个基于比较医学的组件；3)一个咨询委员会；以及4)一个基于约翰霍普金斯医学院的普通组件。研究导师将由心脏病专家查尔斯·洛文斯坦博士提供，他是一氧化氮(NO)和无合成酶研究方面的专家。特别是，他的实验室探索了一氧化氮调节血管炎症的机制。 我的具体研究建议旨在确定NO在调节血小板颗粒胞吐中的作用。特别是，我对N-乙基马来酰亚胺敏感因子(NSF)分子没有任何修饰感兴趣，NSF是一种参与调节多种细胞类型胞吐的分子。我的目的是要证明：1)NO调节血小板颗粒的胞吐；2)NO在体外和体内调节NSF的活性；3)NSF的多肽抑制剂在体外抑制NSF，从而抑制体内血栓形成。 血管炎症是许多心血管疾病的重要组成部分。血小板衍生的血栓前和炎症介质，如在血小板颗粒中发现的那些，是促进血小板和白细胞(特别是中性粒细胞)定位于血管损伤部位的关键，并可能加剧损伤。因此，减少颗粒胞吐的策略在血栓前疾病的急性治疗中可能被证明是有价值的。(摘要结束)

DESCRIPTION (provided by applicant): I am applying for a Mentored Clinical Scientist Career Award (K08) in order to gain mentored basic research training. I wish to investigate nitric oxide regulation of platelet granule exocytosis as it relates to thrombosis. Through this award I seek to become an investigator capable of applying my knowledge and training through 1) a mentored laboratory-based component to acquire knowledge in (a) exocytosis research and (b) thrombosis research; 2) a comparative medicine-based component; 3) an advisory committee; and 4) a general Johns Hopkins Medical Institute-based component. Research mentorship will be provided by Dr. Charles Lowenstein, a cardiologist and an expert in nitric oxide (NO) and NO synthase research. In particular, his laboratory explores mechanisms by which NO regulates vascular inflammation. My specific research proposal is aimed at defining the role of NO in regulating platelet granule exocytosis. In particular, I am interested in NO modification of the molecule N-ethylmaleimide sensitive factor (NSF), a molecule involved in regulating exocytosis in many cell types. My specific aims are intended to demonstrate that: 1) NO regulates platelet granule exocytosis: 2) NO regulates NSF activity in vitro and in vivo; and 3) peptide inhibitors of NSF inhibit NSF in vitro and therefore inhibit thrombosis in vivo. Vascular inflammation is an important component of many cardiovascular diseases. Platelet-derived prothrombotic and inflammatory mediators, such as those found in platelet granules, are key in promoting localization of platelets and leukocytes (neutrophils, in particular) to the sites of vascular injury and may exacerbate the injury. Therefore, strategies to minimize granule exocytosis may prove valuable in the acute treatment of pro-thrombotic diseases. (End of Abstract)