Developing Extracellular Vesicle Based MPRINT Translational Resource Platform for Monitoring Therapeutics Response During Pregnancy

摘要 孕妇和哺乳期的人仍然是治疗孤儿，因为他们被排除在绝大多数 临床药物开发和治疗试验。此外，目前在妊娠期药物评价方面的做法 由于缺乏有效的生物标志物和创新的研究设计，有必要 开发新的胎盘特异性生物标志物以评估胎盘功能和对治疗剂的反应， 作为一种告知其安全性和有效性的方式。这些新的生物标志物的一个例子是胎盘（胎儿） 特异性细胞外囊泡（EV）。在表征这些电动汽车的货物内容方面的最新进展 证明了它们作为胎盘生物标志物的潜力。我们已经展示了使用来自 MPRINT显示，EV中发现的14种蛋白质与高危人群中阿司匹林的使用显著相关（FDR<0.1）， 但需要更多的力量来自信地评估与阿司匹林剂量的关系， 妊娠结局。此外，先前的研究表明阿司匹林影响内皮细胞和滋养层细胞， 从而潜在地调节来源于这些细胞的外泌体及其货物内容物。利用我们的前任 原则成功的证明，先前收集的，以及正在从高危人群中收集的母体血浆 在俄亥俄州，每天服用81 mg或162 mg阿司匹林的先兆子痫（PE）患者和低风险人群中， 大学，我们提交这个应用程序的总体目标是开发一个新的平台，以增加 MPRINT资源使用外泌体分析，作为新的生物标志物，以监测胎盘介导的不良反应， 妊娠结局和对治疗的反应。为此，我们将使用PE作为这些结果的标志 和阿司匹林作为治疗剂，以显示该平台的实用性。在这项研究中，我们将测试 电动汽车蛋白质组货物差异表达与胎盘和妊娠相关的假设 健康的反应阿司匹林治疗在不同的妊娠期。我们将测试以下具体目标： 1)使用来自其他生物库样本的更大队列来验证初步蛋白质组数据和模型， 接受81 mg或162 mg阿司匹林的高危人群，表征胎儿特异性EV（胎盘碱性 磷酸酶[PLAP] +ve），并确定从母体血浆中分离的磷酸酶之间的差异。 用阿司匹林治疗的母体与胎儿EV货物差异;和2）确定与 前瞻性接受阿司匹林的高危人群中EV蛋白质组谱（母体和胎儿）与 临床结局（PE的发生）以及与PE相关的血管生成和炎症生物标志物。这 该项目有可能在开发治疗研究的新平台方面发挥开创性作用 怀孕期间这个转化研究平台将大大增加MPRINT网络的曲目， 通过团队与其他成员之间已经建立的关系， 网络，如产妇胎儿医学单位网络，MPRINT和NIH基金会。

ABSTRACT Pregnant and lactating people remain therapeutic orphans as they are excluded from the vast majority of clinical drug development and therapeutic trials. Moreover, current practices in drug evaluation in pregnancy have been hindered by the lack of effective biomarkers and innovative study designs. There is a need to develop novel placental-specific biomarkers in order to assess placental function and response to therapeutics, as a way to inform on their safety and efficacy. An example of these novel biomarkers is placental (fetal) specific extracellular vesicles (EVs). Recent advances in characterizing the cargo content of these EVs demonstrated their potential to be used as placental biomarkers. We have shown using funding support from the MPRINT that fourteen proteins found in EVs significantly correlated with aspirin use (FDR<0.1) in at-risk people, but that more power is needed to confidently assess the relationship with aspirin dosage and pregnancy outcomes. In addition, prior studies showed that aspirin affects endothelial and trophoblast cells, thus potentially modulating exosome derived from these cells and their cargo contents. Leveraging our prior proof of principle success, previously collected, and ongoing collection of maternal plasma from people at-risk of preeclampsia (PE) receiving 81mg or 162mg of aspirin daily and low-risk people at the Ohio State University, we are submitting this application with the overarching goal to develop a novel platform to augment the MPRINT resources using exosome profiling, as novel biomarkers, to monitor placental mediated adverse pregnancy outcomes and response to therapeutics. For this, we will use PE as a hallmark of these outcomes and aspirin as the therapeutic agent to show case the utility of the platform. In this study, we will test the hypothesis that differential expression of EVs proteome cargo is associated with placental and pregnancy health in response to aspirin treatment at different gestational periods. We will test the following specific aims: 1) Validate our preliminary proteome data and model using larger cohort from additional biobanked samples of at-risk people receiving 81mg or 162 mg aspirin, characterize fetal specific EVs (placental alkaline phosphatase [PLAP] +ve) isolated from maternal plasma, and determine the differences in between the maternal vs. fetal EV cargo difference with aspirin treatment; and 2) Determine the changes associated with EV proteome profile (maternal and fetal) in at-risk people receiving aspirin prospectively and correlate with clinical outcomes (development of PE) and angiogenic and inflammatory biomarkers associated with PE. This project has the potential to play a seminal role in the development of a novel platform for therapeutics research in pregnancy. This translational research platform will substantially add to the MPRINT Network repertoire and be available for scientific community through the already established relationship of the team with other networks such as the Maternal Fetal Medicine Units Network, MPRINT, and the Foundation of the NIH.