The role of Akt in platelet signaling and thrombosis

描述（由申请人提供）： 我的长期职业目标是在一个主要的研究机构获得一个教师职位，那里的资源可以支持糖尿病血管并发症的研究专业。我的短期目标是利用这个研究事业资助奖支持的项目，既发展作为一名研究者的独立性，又扩大我在信号转导和血小板生物学方面的专业知识，包括对血小板在体内作用的更全面的了解。Brass博士在宾夕法尼亚大学医学系Hem-Onc分部的实验室为管理科学和职业过渡提供了理想的环境。科学家，每周数据俱乐部和研讨会系列由当地专家在血小板生物学，糖尿病，血液学和药理学提供活跃的论坛，讨论目前的发展，在每个专业，其中许多导致富有成效的合作。关于我的职业发展，我的赞助商和医学主席都致力于支持我在3年内过渡到正式教师职位。本提案的科学目的是了解调节血小板活化的细胞内信号传导事件，以更好地了解其对血栓形成的贡献。在以下3个特定目的中，本提案将检验Akt 1和Akt 2在导致血小板聚集和体内形成的血小板栓稳定的信号通路中发挥关键作用的假设。 目的1）Akt在体外是否参与血小板活化和聚集？这些研究将继续我已经开始的工作，以确定缺乏Akt 1和Akt 2的多个等位基因的小鼠是否在纤维蛋白原结合，颗粒分泌或凝块收缩方面存在缺陷。 目的2）Akt是如何参与血小板活化的信号通路的？由于这些事件与血小板聚集相关，因此将测试血小板Akt的低表达对GTCRap 1活化和整联蛋白α IIb β 3的β 3尾磷酸化的影响。 目的3）Akt是否参与体内血小板栓的形成？我们将测量出血时间、颈动脉氯化铁损伤后的血流量以及Akt缺陷小鼠与野生型小鼠的血栓栓塞，以确定Akt对止血和血栓形成的影响。

DESCRIPTION (provided by applicant): My long-term career objectives are to obtain a faculty position at a major research institution, where the resources exist to support a research specialty in the vascular complications of diabetes. My short-term goals are to use the project supported by this research career grant award both to develop increasing independence as an investigator and to broaden my developing expertise in signal transduction and platelet biology to include a more complete understanding of the role of the platelet in vivo. The laboratory of Dr. Brass here in the Hem-Onc Division of the Department of Medicine at Penn provides an ideal environment in which to manage both the scientific and career transition. Scientifically, weekly data clubs and seminar series run by local experts in platelet biology, diabetes, hematology, and pharmacology provide lively forums for discussion of the current developments in each specialty, many of which lead to fruitful collaborations. With regard to my career development, both my sponsor and the Chair of Medicine are committed to supporting my transition to a full faculty position within 3 years. The scientific objective of this proposal is to understand the intracellular signaling events that regulate platelet activation to gain a better understanding of their contribution to thrombosis. In the following 3 Specific Aims, this proposal will test the hypothesis that Akt1 and Akt2 play a critical role in the signaling pathways that lead to platelet aggregation and stabilization of the platelet plug formed in vivo. Aim 1) Does Akt contribute to platelet activation and platelet aggregation in vitro? These studies will continue work that I have already begun to determine whether mice lacking multiple alleles of Akt1 and Akt2 have defects in fibrinogen binding, granule secretion, or clot retraction. Aim 2) How is Akt involved in signaling pathways leading to platelet activation? The effects of low expression of platelet Akt on activation of the GTPase Rap1 and phosphorylation of the beta 3 tail of integrin alpha lIb beta 3 will be tested since these events have been associated with platelet aggregation. Aim 3) Does Akt contribute to platelet plug formation in vivo? We will measure bleeding time, blood flow after ferric chloride injury of the carotid artery, and thromboembolism in Akt-deficient versus wildtype mice to determine the effect of Akt on hemostasis and thrombosis.