Model system of oral contraceptive-induced VTE: integrating genomic, transcriptomic, and proteomic discovery with functional biology

静脉血栓栓塞症(VTE)是由血液高凝、血管内皮细胞 调节失调和血瘀证。这种生物失调具有环境和遗传来源 相互作用以调节血栓形成的风险。女性性激素为主的口服避孕药(OCS)是一种重要的 环境因素导致年轻女性血栓形成，使患VTE的风险增加2至4倍。这样做的目的是 应用是为了阐明外源性女性性激素(OCS)增加高血压风险的机制 绝经前妇女的静脉血栓形成。我们的提案有四个目标，这将推动生物发现和功能 实验。R61概述：发现。在此提案的前两年，我们建议确定新的 与OC使用和VTE风险相关的基因、蛋白质标记物和生物途径 资源(基因组、转录组和蛋白质组)和功能细胞生物学和生物化学技术。 目的1：利用OCS在3口井中发现与绝经前妇女静脉血栓形成有关的新基因。 VTE的特征性病例对照研究。我们已经实现了这一目标。目标2：描述以下方面的影响 OCS对内皮细胞(EC)转录和蛋白质组反应、EC促凝活性和 促进血栓形成。我们已经实现了这一目标。R33概述：验证和功能测试。 在这项提案的未来3年里，我们将验证在R61阶段确定的候选基因和蛋白质， 在基于细胞的体外凝血模型中表征它们的功能。我们将描述交互的特征 血管内皮细胞失调与血浆高凝状态之间促进血栓前凝块形成的关系 血液-内皮界面模型。目标3：确定人类群体中的候选基因和蛋白质， 重点关注目标2的发现。目的4：鉴定通过发现和鉴定的新候选基因 验证上述目标，并确定每个基因对正常和 高凝血浆。总而言之，这些目标将发现和功能分析信息集成到 确定促进OC诱导的VTE的生物变异。这项研究将产生功能验证的分子 信号和用于纳入VTE风险预测工具的血液-内皮界面模型 绝经前的女性。

Venous thromboembolism (VTE) is caused by a combination of blood hypercoagulability, endothelial dysregulation, and blood stasis. This biologic dysregulation has environmental and inherited sources that interact to modulate thrombotic risk. Female sex-hormone-based oral contraceptives (OCs) are an important environmental cause of thrombosis in young women, increasing the risk of VTE 2 to 4-fold. The aim of this application is to elucidate mechanisms by which exogenous female sex hormones (OCs) increase the risk of VTE in premenopausal women. Our proposal has 4 aims, which will drive biologic discovery and functional experimentation. R61 Overview: Discovery. In the first 2 years of this proposal, we proposed to identify new genes, protein markers, and biologic pathways associated with OC use and VTE risk using “panomic” resources (genomic, transcriptomic, and proteomic) and functional cell biology and biochemistry techniques. Aim 1: Identify novel genes that contribute to VTE in premenopausal women using OCs in 3 well- characterized, case-control studies of VTE. We have accomplished this Aim. Aim 2: Characterize effects of OCs on endothelial cell (EC) transcriptomic and proteomic responses, EC procoagulant activity, and ability to promote clot formation. We have accomplished this Aim. R33 Overview: Validation and Functional Testing. In the next 3 years of this proposal, we will validate candidate genes and proteins identified in the R61 phase, characterizing their function in an in vitro cell-based model of coagulation. We will characterize interactions between EC dysregulation and plasma hypercoagulability to promote the formation of prothrombotic clots in a blood-endothelial interface model. Aim 3: Characterize candidate genes and proteins in human populations, focusing on Aim 2 discoveries. Aim 4: Characterize the novel candidate genes identified through discovery and validation in the aims above, and determine effects of each gene on EC procoagulant activity in normal and hypercoagulable plasmas. Collectively, these aims integrate discovery and functional-analysis information to identify biologic variation that promotes OC-induced VTE. This study will yield functionally-validated molecular signals and a blood-endothelial interface model for incorporation into VTE risk prediction tools in premenopausal women.