Quantitative proteomic profiling of extracellular vesicles from menstrualeffluent for the discovery of non-invasive diagnostic biomarkers of endometriosis

摘要： 子宫内膜异位症是一种子宫内膜腺和间质向宫腔外延伸而引起的疾病。 大约10%的育龄妇女，仅在美国就有400多万人受到 子宫内膜异位症；由于未确诊病例的高比率，真实的患病率预计要高得多。 已经有记录表明，患有 子宫内膜异位症。尽管有巨大的兴趣和未得到满足的需求，但没有FDA批准的非侵入性 子宫内膜异位症的诊断工具或生物标志物。其中一个关键原因是缺乏对 临床症状、组织学表现与潜在功能障碍的相关性 疾病。此外，子宫内膜异位症通常与一系列其他疾病有关，从而产生高生物标记物。 噪音。细胞外小泡(EV)是由所有细胞分泌的，被认为是介导细胞间以及 器官间的沟通和已知的沟通特定的疾病状态。电动汽车已经成为 推测在子宫内膜异位症的发病机制和诊断中起一定作用。在疾病中，EV含量是已知的 以高度特异的方式改变以反映疾病类型。我们试图克服低信号、高噪音 子宫内膜异位症生物标记物发现的障碍：1)专注于EVS，这种疾病的细胞信使， 存在于月经流出物中2)使用最先进的、高度敏感的定量蛋白质组技术，以及3) 使用生物标志物的队列/小组。我们在蛋白质组学技术上的改进就导致了10倍以上的 显著的数量差异。它们结合在一起，提供了非常重要的概念、技术和 仪器仪表超越了过去的研究。我们将使用等压进行多路定量蛋白质组学 经手术证实的子宫内膜异位症患者月经流出物的串联质量标记(TMT) 将他们与有类似子宫内膜异位症症状的对照组进行比较，包括慢性盆腔疼痛，但没有 在手术中检测到子宫内膜异位症。它将为我们提供患者特有的、高质量的数据库 月经流出液中分泌的外切体的蛋白质组内容，并鉴定其上的功能信号网络， 为从月经流出液中开发电动汽车奠定基础。这笔赠款的目标是确定 月经流出物EVS上的子宫内膜异位症生物标志物的队列/小组，可用于检测 第二阶段的开发和临床验证。

Abstract: Endometriosis is a disease caused by the extension of endometrial glands and stroma outside the uterine cavity. Approximately 10% of reproductive age women, more than 4 million in the United States alone, are affected by endometriosis; the true prevalence is expected to be much higher due to the high rate of undiagnosed cases. Significant physical, psychological, and economic suffering has been documented in patients suffering from endometriosis. In spite of the tremendous interest and unmet need, there is no FDA-approved non-invasive diagnostic tool or biomarker for endometriosis. One of the key reasons is the lack of understanding of the correlation between clinical symptoms, histological manifestations, and underlying dysfunction resulting in disease. In addition, endometriosis is often associated with a range of other disorders, creating a high biomarker noise. Extracellular vesicles (EV) are secreted by all cells and are thought to mediate intercellular as well as inter-organ communication and are known to communicate the specific disease state. EVs have been hypothesized to play a role in the pathogenesis and diagnosis of endometriosis. In disease, EV content is known to be altered to reflect the disease type in a highly specific way. We seek to overcome the low signal, high noise hurdle of endometriosis biomarker discovery by 1) focusing on EVs, the cellular messengers of the disease, present in menstrual effluent 2) using state of the art, highly sensitive, quantitative proteomic technique, and 3) using cohort/panel of biomarkers. Our improvement in proteomic technique alone results in 10-fold more significant quantitative differences. In combination, they present a very significant conceptual, technical, and instrumentation leap over past studies. We will perform the multiplexed quantitative proteomics using isobaric tandem mass tags (TMT) on menstrual effluent EVs from surgically confirmed endometriosis patients and compare them to controls with endometriosis like symptoms including chronic pelvic pain, but with no endometriosis being detected at the surgery. It will provide us a patient specific, high-quality database of proteomic content of exosomes secreted in menstrual effluent and identify functional signaling networks on them, to lay down the foundation for the exploration of EVs from menstrual effluent. The goal of this grant is to identify the cohort/panel of biomarkers of endometriosis on menstrual effluent EVs, that can be taken to assay development and clinical validation in phase II.