OPA1 an Estrogen-Mediated Modulator of Platelet Hyperactivation

OPA1 是雌激素介导的血小板过度活化调节剂

基本信息

批准号：
10026343
负责人：
E Dale Abel
金额：
$ 72.43万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-04-10 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10026343
关键词：
Address Animal Model Animals Bioenergetics Biological Models Blood Platelets Collaborations Complement Complex Contraceptive Usage Coronary Arteriosclerosis Coronary heart disease Data Diabetes Mellitus Epidemiology Estrogen Replacements Estrogens Exhibits Exposure to Female Follow-Up Studies Gene Expression Gonadal Steroid Hormones Hemostatic Agents Hormones Human Impairment In Vitro Inner mitochondrial membrane Iowa Knock-out Knockout Mice Knowledge Link Mediating Megakaryocytes Membrane Proteins Messenger RNA Metalloproteases Mitochondria Modeling Molecular Morphology Mus OPA1 gene Optic Atrophy Oral Contraceptives Ovariectomy Participant Patients Pattern Peptide Hydrolases Phase Phenotype Physiological Platelet Activation Platelet aggregation Postpartum Period Pregnancy Pregnant Women Premenopause Proteins Recording of previous events Risk Role Signal Pathway Signal Transduction System Testing Third Pregnancy Trimester Thrombosis Time Transcript Transgenic Mice Universities Utah Venous Thrombosis Woman Work Zinc base cohort genetic manipulation human model human pluripotent stem cell human subject in vivo in vivo Model indexing induced pluripotent stem cell innovation male metabolomics molecular phenotype mouse model next generation novel novel marker overexpression pill platelet function platelet phenotype pregnant programs recruit response sex transcriptome sequencing transcriptomics venous thromboembolism

项目摘要

Strong epidemiological evidence links pre-menopausal estrogen exposure to increased risk of venous thrombosis in women, particularly during pregnancy, in the immediate postpartum period, and in women taking estrogen-containing oral contraceptives (OCP). This proposal addresses a significant knowledge gap linking estrogens with platelet activation. Studies in the past have focused on how estrogens alter the hemostatic and fibrinolytic system. This proposal, which focuses on platelets, represents a productive collaboration between the Universities of Iowa and Utah. Our preliminary studies in the Framingham cohort and in pregnant humans, reveal increased platelet OPA1 levels in females, that predict risk of coronary artery disease, or correlate with increased platelet activation in the third trimester of pregnancy. In platelet-deficient OPA1 knockout (KO) mice we discovered that female OPA1KO mice were protected from thrombosis compared to males. The thrombosis phenotype was reversed after oophorectomy. Based on these preliminary findings, we hypothesize that OPA1 and regulators of OPA1 activity, OMA1 and YMEL1, modulate platelet activation in an estrogen-dependent manner and OPA1 levels in platelets predict estrogen modulated platelet activation. To address this hypothesis in the R61 phase, Aim 1 will determine the molecular mechanisms for estrogen-mediated and OPA-1-dependent modulation of platelet activation using mice with platelet specific OPA1 deletion and will generate OMA1 and YMEL1 platelet specific KO mice, with increased OPA1 activity. Aim 2 will determine platelet OPA1, OMA1, and YMEL1 expression and correlate expression levels to platelet activation in the setting of pregnancy and OCP use. We will also develop a human iPSC model system to generate MKs to determine if estrogen directly regulates the expression of these genes. The R33 phase will dissect the molecular mechanisms linking increased OPA1 with platelet activation in females. Aim 1 will perform detailed physiological and molecular phenotyping of murine platelets isolated from OMA1 and YMEL1 KO and OPA overexpressing transgenic mice. In addition, we will examine the mechanisms linking increased OPA1 activity, mitochondrial energetics and platelet activation. Aim 2 will complement animal studies by examining changes in mitochondrial function and signaling in platelets from pregnant women and women on OCPs. We will genetically ablate OMA1 and YMEL1 from iPSC-derived MKs to directly test if estrogen modulates MK and platelet function in an OPA1 dependent manner. By integrating in vivo and in vitro studies in humans and mouse models, our approach is translational, innovative and highly responsive to the RFA. These studies will determine for the first time whether increased estrogen exposure in pregnancy and OCP use reprogram MKs and developing platelets to increase thrombotic complications and identify novel mechanisms for this increased risk. The impact of these studies, is that they may identify novel biomarkers that may predict increased risk of thrombosis in susceptible females during pregnancy or prior to OCP use.

强有力的流行病学证据将绝经前雌激素暴露与静脉风险增加联系女性的血栓形成，特别是在怀孕期间，产后立即怀孕，服用女性含雌激素的口服避孕药（OCP）。该建议解决了一个重大的知识差距链接血小板激活的雌激素。过去的研究集中于雌激素如何改变止血和纤维蛋白水解系统。该提案专注于血小板，代表了爱荷华州和犹他州的大学。我们在Framingham队列和怀孕人类的初步研究，揭示女性血小板OPA1水平升高，预测冠状动脉疾病的风险或与妊娠三个月的血小板激活增加。在缺乏血小板的OPA1敲除（KO）小鼠中我们发现与男性相比，雌性Opa1ko小鼠免受血栓形成。这卵巢切除术后血栓形成表型逆转。基于这些初步发现，我们假设 OPA1活性OMA1和YMEL1的OPA1和调节剂调节血小板激活雌激素依赖性方式和血小板中的OPA1水平预测雌激素调节血小板激活。为了解决R61阶段的这一假设，AIM 1将确定分子机制雌激素介导的和OPA-1依赖性调节血小板激活，使用具有血小板特异性的小鼠 OPA1缺失并将产生OPA1活性增加的OMA1和YMEL1血小板特异性KO小鼠。 AIM 2将确定血小板OPA1，OMA1和YMEL1表达，并将表达水平与血小板相关在怀孕和OCP使用的情况下激活。我们还将开发人类IPSC模型系统生成MK来确定雌激素是否直接调节这些基因的表达。 R33阶段将剖析连接的分子机制增加了OPA1与女性的血小板激活。目标1意志从OMA1和YMEL1分离出的鼠血小板的详细生理和分子表型 KO和OPA过表达的转基因小鼠。此外，我们将检查连接的机制增加 OPA1活性，线粒体能量和血小板激活。 AIM 2将通过检查孕妇和妇女血小板中线粒体功能的变化和信号的变化 OCPS。我们将从IPSC衍生的MK中遗传渗透OMA1和YMEL1，以直接测试雌激素是否存在以OPA1依赖性方式调节MK和血小板功能。通过整合体内和体外研究在人类和老鼠的模型中，我们的方法是转化，创新性和对RFA的高度响应。这些研究将首次确定怀孕和OCP使用中的雌激素暴露是否增加重新编程MK和开发血小板以增加血栓形成并发症并确定新型机制因为这增加了风险。这些研究的影响是，他们可以识别出可能预测的新型生物标志物怀孕期间或使用OCP之前，易感女性血栓形成的风险增加。