Electrochemical Liquid Biopsy Assessing Placental Health

胎盘健康对孕期健康至关重要，对母亲和子女的终生都有影响 健康。人们认识到，胎盘着床、细胞发育和/或成熟过程中的异常 对母胎营养供应产生不利影响，对发育中的新生儿产生不利影响。 目前可用于评估胎盘健康状况的临床工具包括超声(U/S)和继发性 对孕妇进行的其他指征的分析，例如胎儿非整倍体；这些包括血清 生物标志物、无创产前检测(NIPT)和侵入性检查绒毛、羊水 液体或脐静脉血。然而，这些诊断模式中的许多(如果不是全部的话)都有局限性 与可预测性中的次优敏感性、特异性和准确性有关，或与侵袭性有关 可能会出现并发症的测试。因此，迫切需要非侵入性和易于部署的诊断 在预测胎盘健康方面表现出高度敏感性和特异性的工具。至少，部署的能力 一项在妊娠早期进行的筛查试验，旨在推荐更精细的成像来评估胎盘健康 是必要的。为此，我们对预期建立的队列的初步数据表明，母亲 可以分析妊娠期间纵向采集的血浆和尿液中cfDNA，cfRNA， 外切体RNA/miRNA，以及来自胎盘的蛋白质。为了克服繁琐的cfDNA分析， CfRNA和exosome RNA/Micro(Mi)RNA/蛋白质通过传统的方法，我们最近发展了一种 基于电磁和电化学生物传感器的技术，称为“电化学液体活组织检查”(ELB)， 用于从体液(例如尿液、唾液)样本中分离和鉴定RNA和蛋白质。这部小说 技术为我们提供了非侵入性研究高表达的关键转录本/蛋白质的能力 随后临床胎盘紊乱的敏感性、特异性和可预测性。基于此集合的 初步数据，我们假设一种非侵入式生物传感器能够快速检测RNAs/miRNAs和 尿液中的蛋白质将允许以可靠的方式纵向检测胎盘健康。如果制造了这样的工具 最终可用于护理点检测可以彻底改变对胎盘健康的实时监测 准确地怀孕，允许及时引入新的干预措施(例如他汀类药物)以预防和从而 终止与胎盘相关的临床疾病。为了验证这一假设，我们提出了两个具体目标：1) 用于分离和同时测定多种转录本/蛋白质的ELB装置的研制和测试 正常妊娠期间纵向收集的尿液。2)确定部署的能力和有效性 电化学生物传感器检测正常和高危人群转录本/蛋白质差异的研究 为将来的护理点检测做准备的怀孕，以实现实时、无创和快速 胎盘健康监测。将尿检结果与配对的具有可预测性的血液样本进行比较验证 通过收集胎盘磁共振成像、临床特征和胎盘组织病理学的档案资料进行评估。

Placental health is essential for the well-being of pregnancy, with implications for mother and offspring's lifelong health. Recognition exists that aberrations in placental implantation, cellular development and/or maturation adversely affect the materno-fetal supply of nutrients with detrimental effects on the developing newborn. Currently the clinical tools available for assessing placental well-being consist of ultrasound (U/S), and secondary analysis of maternal tests performed for other indications, such as fetal aneuploidies; these include serum biomarkers, non-invasive prenatal testing (NIPT), and invasive procedures accessing chorionic villi, amniotic fluid or umbilical venous blood. However many if not all of these diagnostic modalities are plagued with limitations either related to suboptimal sensitivity, specificity and accuracy in predictability, or related to the invasive nature of testing with possible complications. Hence, there is a dire need for non-invasive and easy to deploy diagnostic tools that portray high sensitivity and specificity in predicting placental health. At a minimum, the ability to deploy a screening test early in 1st trimester towards recommending a more elaborate imaging to assess placental health is necessary. To this end, our preliminary data on a prospectively established cohort demonstrated that maternal plasma and urine collected longitudinally through pregnancy can be analyzed for a combination of cfDNA, cfRNA, exosome RNA/miRNA, and proteins, arising from placenta. To overcome the cumbersome analysis of cfDNA, cfRNA and exosome RNA/micro(mi)RNA/proteins by traditional methods, we have most recently developed an electromagnetic and electrochemical biosensor based technology named “electrochemical liquid biopsy” (eLB), for separation and identification of RNAs and proteins from bodily fluid (e.g. urine, saliva) samples. This novel technology provides us the ability in non-invasively investigating key transcripts/proteins that display high sensitivity, specificity and predictability of subsequent clinical placental disorders. Based on this collection of preliminary data, we hypothesize that a non-invasive biosensor capable of rapidly detecting RNAs/miRNAs and proteins in urine will allow longitudinal detection of placental health in a reliable manner. Such a tool if made ultimately available for point-of-care testing can revolutionize monitoring of placental health in real-time during pregnancy with accuracy, allowing timely introduction of novel interventions (e.g. statins) to prevent and thereby terminate placenta-associated clinical disorders. To test this hypothesis we propose two specific aims: 1) To develop and test an eLB device in separation and simultaneous measurement of multiple transcripts/proteins in urine collected longitudinally during normal pregnancies. 2) To determine the ability and validity in deploying the electrochemical biosensor tool in detecting transcript/protein differences between normal and high-risk pregnancies in preparation for future point-of-care testing in achieving real-time non-invasive and rapid monitoring of placental health. Validation by comparing urinary results to paired blood samples with predictability assessed by the archived collection of placental MR imaging, clinical features and placental histopathology.