A SYSTEMS BIOLOGY APPROACH TO IDENTIFYING THE MECHANISMS OF SEX HORMONE INDUCED THROMBOEMBOLISM IN PRE-MENOPAUSAL WOMEN

口服避孕药（OC）等激素治疗会增加静脉血栓栓塞（VTE）的风险， 绝经前妇女在我们获得关于为什么会发生这种情况的机械见解之前， 有性侵犯诱发静脉血栓栓塞风险的人本研究的长期目标是确定 性激素调节止血和血栓形成。本申请的总体目标是 使用结合组学技术的系统生物学方法确定OC如何改变血小板功能 用计算机模型。先前对血栓诱发静脉血栓栓塞的研究涉及到几种机制 与血小板功能有关，包括对激动剂的反应、花生四烯酸（AA）的代谢和基因 表情系统地研究这些过程在不同的时间尺度上以及它们之间的关系 这是不可能的，我们将在这里讨论。中心假设是，OC增加血小板反应性超过3 时间尺度;（i）通过增强细胞内储存的钙释放急性（秒-分钟），（ii） 代谢上（分钟-小时）通过升高血栓素代谢，和（iii）基因组上（天-月）通过 改变粘附受体的表达。这一假设是基于初步数据， 与生理浓度的17β-雌二醇孵育的细胞内钙浓度较高 随后粘附到胶原蛋白上并改变了中枢代谢。建议研究的理由二- 折叠：（i）开发新的工具，在多个时间尺度上研究血栓引起的VTE，以及（ii） 在这些时间尺度上测量外源性激素对血小板功能的影响。这一假设将 通过两个具体目标进行测试：1）开发系统生物学工具。2)测量OC对 不同时间尺度的血小板功能。在第一个目标下，钙动力学的计算模型和 通过血小板粘附和代谢流实验，了解血小板中的代谢 分析.此外，我们将对已知影响血小板功能的基因变异进行测序， 激素受体最后，现有的血管损伤微流体模型将被完善， 内皮细胞来测量血小板-内皮相互作用。在第二个目标下，我们将使用工具 开发的第一个目的是测量急性和慢性暴露后血小板功能的变化 外源激素这些研究将包括跟踪一组妇女在接受治疗前和治疗后的血小板功能。 开始OC后这一方法是创新的，因为它代表了一种新的、实质性的背离， 通过使用系统生物学方法来测量和模拟性激素对 血小板功能在几秒到几年的时间尺度上变化。这项研究意义重大，因为它将 确定外源性性激素赋予促血小板和/或抗血小板表型的机制， 绝经前妇女通过非基因组和基因组途径。最终，这些知识 有可能提高美国激素治疗的安全性。

Hormone therapies like oral contraception (OC) confer a heightened risk of venous thromboembolism (VTE) in premenopausal women. Until we gain mechanistic insights into why this happens, it is not possible to predict who is at risk for sex hormone-induced VTE. The long-term goal of this research is to identify the mechanisms by which sex hormones modulate hemostasis and thrombosis. The overall objective in this application is to determine how OC alter platelet function using a systems biology approach that combines –omics technologies with computational models. Previous work on hormone-induced VTE have implicated several mechanisms related to platelet function including response to agonists, metabolism of arachidonic acid (AA), and gene expression. Systematic studies of these processes over different time scales and how they relate to each other is lacking and will be addressed here. The central hypothesis is that OC increase platelet reactivity over three time scales; (i) acutely (seconds-minutes) by potentiating calcium release from intracellular stores, (ii) metabolically (minutes-hours) by elevating thromboxane metabolism, and (iii) genomically (days-months) by altering the expression of adhesive receptors. This hypothesis is based on preliminary data that platelets incubated with physiologic concentrations of 17β-estradiol have higher intracellular calcium concentrations following adhesion to collagen and altered central metabolism. The rationale for the proposed research two- fold: (i) the development of new tools to study hormone-induced VTE over multiple time scales, and (ii) to measure the effects of exogenous hormones on platelet function over these time scales. This hypothesis will be tested by two specific aims: 1) Development of systems biology tools. 2) Measuring the effects of OC on platelet function over diverse time scales. Under the first aim, computational models of calcium dynamics and metabolism in platelets will be developed informed by experiments of platelet adhesion and metabolic flux analysis. Additionally, we will perform sequencing of gene variants known to affect platelet function and hormone receptors. Finally, existing microfluidic models of vascular injury will be refined to incorporate endothelial cells to measure platelet-endothelium interactions. Under the second aim, we will use the tools developed in the first aim to measure changes in platelet function following acute and chronic exposure to exogenous hormones. These studies will include tracking platelet function in a cohort of women prior to and after starting OC. The approach is innovative because it represents a new and substantive departure from the status quo by using a systems biology approach to measure and model the influence of sex hormones on platelet function over time scales of seconds to years. The proposed research is significant, because it will identify the mechanism(s) by which exogenous sex hormone confer a pro- and/or antiplatelet phenotype in premenopausal women by both non-genomic and genomic pathways. Ultimately, such knowledge has the potential to improve the safety of hormone therapies in the United States.