Molecular profiling of the platelet thrombohemorrhagic

描述（由申请人提供）： 血小板参与缺血性病变的演变，并且通常起保护作用以免受损伤的出血后果;此外，抗血小板剂是心血管和脑血管疾病治疗的主要支柱。尽管有这些基本的重要功能，但关于调节血栓出血性血小板表型的血小板基因和蛋白质的信息很少。最近，我们的实验室已经证明，转录谱技术（在遗传解剖和恶性肿瘤的预测模型中发展良好）可以应用于血小板，并进行了必要的修改，旨在解决RNA产量和白细胞污染的限制。最近，我们已经将这种方法应用于原发性血小板增多症（ET）的研究，这是一种经常与血栓出血性后果相关的血小板疾病。由于与ET相关的血栓性表型是血小板限制性的，我们建议进一步发展这一主题，作为一个范例，用于识别血小板相关的分子签名，可能是血栓性或出血性中风的因果关系。在这个探索性的R21提案中，我们现在建议将我们在血小板转录组学方面的专业知识吸收到蛋白质组学的扩展研究重点中，其长期目标是最佳地定义调节血栓出血平衡的血小板相互作用网络。一个在计算生物学、遗传学、止血和蛋白质组学方面具有相当专业知识的多学科团队已经组装起来专门开发这一主题。在具体目标1中，我们建议开发一个强大的集成遗传和蛋白质组血小板分析平台;该重点的子目标是开发一个基于网络的、完全注释的界面，供对集成血小板蛋白质组感兴趣的广泛研究界感兴趣/转录组分析。在具体目标2中，我们将描述和表征区分血栓/出血表型的血小板基因和蛋白质的初始类别。我们在功能和分子血小板分析方面的集体经验和综合平台使我们的团队处于独特的位置，可以完成这些研究。虽然我们最初的研究集中在一种罕见的血小板疾病（即ET）的转录组学分析，但在本提案的背景下提出的综合蛋白质组学研究可能会对更大的脑血管或心血管疾病患者子集产生更广泛的影响，从而扩大研究方向。

DESCRIPTION (provided by applicant): Platelets are involved in the evolution of ischemic lesions, and normally function to protect from the hemorrhagic consequences of injury; furthermore, antiplatelet agents are the mainstays of treatment in cardiovascular and cerebrovascular disease. Despite these fundamentally important functions, very little information is available on the platelet genes and proteins that modulate the thrombohemorrhagic platelet phenotype. Recently, our laboratory has demonstrated that transcript profiling techniques (well-developed in genetic dissection and predictive models of malignancy) can be applied to platelets, with uniquely-developed modifications aimed at addressing limitations of RNA yield and leukocyte contamination. More recently, we have applied this approach to the study of essential thrombocythemia (ET), a platelet disorder frequently associated with thrombohemorrhagic consequences. Since the thrombohemorrhagic phenotypes associated with ET are platelet-restricted, we propose to further develop this theme as a paradigm for identification of platelet-related molecular signatures that may be causally implicated in thrombotic or hemorrhagic stroke. In this exploratory R21 proposal, we now propose to assimilate our expertise in platelet transcriptomics to an expanded research focus in proteomics, with the long-term goal of optimally defining platelet interactive networks that regulate the thrombohemorrhagic balance. A multidisciplinary team with considerable expertise in computational biology, genetics, hemostasis, and proteomics has been assembled to specifically develop this theme. In specific aim 1, we propose to develop a robust platform for integrated genetic and proteomic platelet analyses; a subaim of this focus will be development of a web-based, fully-annotated interface of interest to the broad research community interested in integrated platelet proteomic/transcriptomic analyses. In specific aim 2, we will delineate and characterize an initial class of platelet genes and proteins that discriminate between the thrombo/hemorrhagic phenotype. Our collective experience and integrated platform for functional and molecular platelet analysis place our group in a unique position to complete these studies as outlined. While our initial studies have focused on a transcriptomic analysis of a rare platelet disorder (i.e. ET), it is likely that integrated proteomic studies proposed within the context of this proposal will have broader implications to the larger subsets of patients with cerebrovascular or cardiovascular disease, leading to an expanded research direction.