Extracellular Vesicles as a Link Between Placental and Renal Dysfunction in Preeclampsia

项目摘要 先兆子痫（PE）是一种毁灭性的妊娠高血压疾病，是孕产妇和新生儿死亡的主要原因。 世界范围内的胎儿死亡率和发病率。它影响2 - 10%的妇女，占孕产妇死亡的16%。 与发达国家的分娩有关。PE很难研究，因为它是一种多因素疾病， 与临床症状谱的发展相关的不同分子机制 与早发、晚发和重度PE相关。个性化医疗方法应用于 剖析PE的潜在机制可能有助于减少这种破坏性的临床发病率， 综合征胎盘在PE的发展中起着关键作用，导致广泛的母体内皮细胞损伤。 PE中的功能障碍、高血压和全身性多器官衰竭。细胞外囊泡（EV），其含有 蛋白质、RNA和脂质被不断地从胎盘中挤出，并能够与胎盘相互作用。 包括肾脏在内的母体器官PE主要与胎盘和肾功能不全相关， 是怀孕期间急性肾损伤的最常见原因。然而，没有研究调查了 胎盘来源的RNA货物作为PE中胎盘和肾功能障碍之间的联系的潜力。尿EV 来源于多种组织类型，代表了越来越多地被利用的生物标志物的宝库 来诊断肾脏疾病此外，可以在整个妊娠期间非侵入性地获得尿样， 可潜在地用于鉴定与PE中胎盘功能障碍相关的生物标志物。拟议 研究，在指导阶段，尖端RNA-Seq技术与计算生物学相结合 和机器学习方法将被应用于分析患有PE的女性的尿液EV的转录组 与正常怀孕相比。初步数据表明，可以分离和分析 在整个正常妊娠期间，来自母体尿的尿EV的转录组，以及胎盘来源的和 胎盘特异性mRNA和miRNA可以在尿EV群体中检测到。这是一部小说 这项技术有可能鉴定生物标志物，并提供有关胎盘功能障碍信息， 以非侵入性方式进行PE。在独立阶段，候选人将利用体外方法， 研究具有与PE相关的miRNA货物的胎盘来源的EV的摄取对功能的影响， 近曲小管上皮细胞和皮质集合管细胞。这两种肾脏特异性细胞类型是 参与肾单位的肾小管重吸收，这是一个导致排泄增加的过程 在一些先兆子痫孕妇的尿液中的蛋白质。该提案是多学科的，利用基本的 生物学、临床研究和高性能计算应用于研究胎盘功能障碍， 体育课这些实验意义重大，因为它们将产生关于胎盘作用的新信息- 衍生EV在PE肾功能不全中的应用，以及为预防和治疗目标指明方向 这可能是变革性的和临床相关的。

PROJECT SUMMARY Preeclampsia (PE) is a devastating hypertensive disorder of pregnancy that is a leading cause of maternal and fetal mortality and morbidity worldwide. It affects 2-10% of women and accounts for 16% of maternal deaths related to childbirth in developed countries. PE is difficult to study because it is a multifactorial disorder with varying molecular mechanisms associated with the development of the spectrum of clinical symptoms associated with early onset, late onset, and severe PE. A personalized medicine approach applied to dissecting the underlying mechanisms of PE may be beneficial to reduce clinical incidence of this devastating syndrome. The placenta plays a key role in the development of PE, leading to widespread maternal endothelial dysfunction, hypertension, and systemic multi-organ failure in PE. Extracellular vesicles (EVs) containing protein, RNA, and lipid cargo are continuously extruded from the placenta, and are capable of interacting with maternal organs including the kidney. PE is primarily associated with placental and renal dysfunction, and PE is the most common cause of acute kidney injury during pregnancy. However, no studies have investigated the potential of placenta-derived RNA cargo as a link between placental and renal dysfunction in PE. Urinary EVs are derived from multiple tissue types and represent a trove of biomarkers that are increasingly being utilized to diagnose renal disorders. Further, urine samples can be obtained throughout pregnancy non-invasively and could potentially be utilized to identify biomarkers related to placental dysfunction in PE. In the proposed research, during the mentored phase, cutting-edge RNA-Seq technology coupled with computational biological and machine learning approaches will be applied to profile the transcriptome of urinary EVs in women with PE compared to normal pregnancy. Preliminary data indicates that it is possible to isolate and profile the transcriptome of urinary EVs from maternal urine throughout normal gestation, and that placenta-derived and placenta-specific mRNA and miRNA can be detected within the urinary EV population. This presents a novel technique that has potential to identify biomarkers as well as provide information on placental dysfunction in PE in a non-invasive manner. During the independent phase, the candidate will utilize an in vitro approach to investigate the effect of uptake of placenta-derived EVs with miRNA cargo associated with PE on the function of proximal tubule epithelial cells and cortical collecting duct cells. These two renal-specific cell types are involved in tubular reabsorption in the nephron, a process that is compromised leading to increased excretion of protein in the urine in some preeclamptic pregnancies. This proposal is multidisciplinary, utilizing basic biology, clinical research, and high-performance computing applied to investigating placental dysfunction in PE. These experiments are significant because they will generate novel information on the role of placenta- derived EVs in renal dysfunction in PE, as well as point the way towards preventative and therapeutic targets that may be transformative and clinically relevant.