Genetic dissection of the platelet thrombohemorrhagic phenotype

描述(申请人提供)：血小板参与缺血性损伤的演变，正常功能是保护免受损伤的出血后果；此外，抗血小板药物是心脑血管疾病的主要治疗药物。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的信息很少。最近，我们的实验室已经证明转录谱技术(在基因解剖和恶性肿瘤预测模型方面发展得很好)可以应用于血小板，其独特的改进旨在解决RNA产量和白细胞污染的限制。最近，我们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后果相关的血小板紊乱。由于与ET相关的血栓出血表型是造血细胞受限的(即血小板和/或白细胞)，我们建议进一步发展这一主题，作为鉴定可能与血栓或出血性中风有关的血小板相关分子特征的范例。我们现在建议吸收我们在血小板分析方面的专业知识，以最佳方式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成立，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综合遗传和蛋白质组血小板分析平台；这一重点的一个子目标将是开发一个基于网络的、完全注释的界面，以吸引对综合血小板蛋白质组/转录组分析感兴趣的广泛研究社区。在特定的目标2中，我们将描述和表征区分血栓/出血表型的一类最初的血小板基因和蛋白。在具体目标3中，我们将开发适用于更大队列的血栓出血的分类预测和评分模型。在这一提议的背景下提出的综合蛋白质组学研究可能会对更多的脑血管或心血管疾病患者产生更广泛的影响，从而扩大未来的研究方向。公共卫生相关性：众所周知，血小板可以调节凝血(血栓形成)和出血(出血)，尽管目前只发现了有限数量的基因来控制这种平衡。在这项提案中，我们将开发和应用复杂的血小板图谱技术来识别可能与出血和血栓形成之间的平衡有关的血小板基因。我们将研究一种人类血小板紊乱症(特发性血小板增多症)作为一个集中的模型系统，当基因被识别时，将该模型应用于扩大的患者队列。这些研究对更多可能患有心血管疾病和/或中风的患者具有广泛的影响。

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 hematopoietic cell-restricted (i.e. platelets and/or leukocytes), 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. We now propose to assimilate our expertise in platelet profiling to optimally define 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. In specific aim 3, we will develop class prediction and scoring models for thrombohemorrhage applicable to larger cohorts. 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 future research direction. PUBLIC HEALTH RELEVANCE: Blood platelets are known to regulate clotting (thrombosis) and bleeding (hemorrhage), although only a limited number of genes have been identified that control this balance. In this proposal we will develop and apply sophisticated platelet profiling technologies to identify platelet genes that may be causally implicated in the balance between hemorrhage and thrombosis. We will study a human platelet disorder (essential thrombocythemia) as a focused model system, and as genes are identified, apply the model to expanded patient cohorts. These studies have wide-spread implications for larger numbers of patients who may suffer from cardiovascular disease and/or stroke.