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

静脉血栓栓塞症（VTE）是由血液高凝状态、内皮细胞损伤和血栓形成引起的。 失调和血液停滞。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风险。女性性激素口服避孕药（OC）是一种重要的 在年轻女性血栓形成的环境原因，增加静脉血栓栓塞的风险2至4倍。的目的 应用是阐明外源性雌性激素（OC）增加风险的机制， 绝经前妇女的VTE。我们的计划有4个目标，这将推动生物发现和功能 实验R61概述：发现。在这项建议的头两年，我们建议确定新的 使用“全基因组”与OC使用和VTE风险相关的基因、蛋白质标记物和生物学途径 资源（基因组学、转录组学和蛋白质组学）以及功能细胞生物学和生物化学技术。 目的1：使用3孔板中的OC鉴定导致绝经前妇女VTE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 OC对内皮细胞（EC）转录组学和蛋白质组学反应、EC促凝血活性和 促进凝块形成。我们已经实现了这一目标。R33概述：确认和功能测试。 在未来3年的计划中，我们将验证R61阶段确定的候选基因和蛋白质， 表征它们在体外基于细胞的凝血模型中的功能。我们将描述 EC失调和血浆高凝状态之间的关系，以促进血栓前凝块的形成， 血液-内皮界面模型目标3：表征人类群体中的候选基因和蛋白质， 专注于Aim 2发现。目的4：表征通过发现和鉴定的新候选基因， 验证上述目的，并确定每个基因对正常和 高凝血浆总的来说，这些目标整合了发现和功能分析信息， 确定促进OC诱导的VTE的生物学变异。这项研究将产生功能验证的分子 信号和血液-内皮界面模型，用于纳入静脉血栓栓塞风险预测工具， 绝经前妇女

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.