High-throughput Integrated Magneto-electrochemical Exosome (HiMEX) platform to identify neurodevelopmental markers associated with pre and postnatal oxycodone exposure

在怀孕期间和怀孕后依赖处方阿片类药物对母亲都构成了重大的健康风险 和孩子.暴露于阿片类药物的新生儿会出现药物戒断综合征， 认知障碍然而，对这些暴露的新生儿的临床护理受到当前 知识的根本差距：缺乏可靠的生物标志物，可客观地评估新生儿的风险， 药物暴露：人们对产前和产后使用阿片类药物如何影响后代知之甚少，特别是 神经发育，没有可靠的生物标志物可用于客观评估新生儿的药物风险， exposure.我们寻求推进一种新的检测平台，以有效监测新生儿对羟考酮的暴露 （氧基）。我们的方法将基于两种创新方法：细胞外囊泡（EV）作为生物标志物 和iMEX（集成磁电化学外泌体）作为传感器平台。目标1.我们将识别EV 暴露在高风险氧气中的蛋白质特征我们将使用啮齿动物模型来模拟子宫内和产后 氧气暴露将收集来自氧暴露后代大脑的EV，并通过定量方法进行分析。 蛋白质组学来鉴定差异表达的蛋白质。目标2.我们将实施第二代iMEX 大大提高了分析能力。我们将通过探索新的方法来提高iMEX检测灵敏度 信号放大（纳米等离子体）;建立一个统一的测定来检测跨膜和 囊内标记物;并构建高通量检测器（96孔板形式）。这个新系统将 用于筛选暴露于氧气的动物的血浆EV。这个项目的成功将产生 关于脑源性EV在氧暴露下的全面蛋白质数据，以及EV作为 生物标记物。此外，所获得的见解将为进一步扩展HiMEX技术的临床应用奠定基础。 验证。我们的长期目标是提供一种微创血液检测，用于风险评估， 氧暴露的干预。

Dependency on prescription opioids during and after pregnancy poses a significant health risk, both to mother and child. Newborns exposed to opioids would experience a drug withdrawal syndrome and have elevated risk of cognitive disorders. Clinical care for these exposed neonates, however, is challenged by a current fundamental gap in knowledge: lack of reliable biomarkers available to objectively assess newborn's risk from drug-exposure: it is poorly understood how pre- and postnatal opioid use affects offsprings, particularly with neurodevelopment, and no reliable biomarkers are available to objectively assess a newborn's risk from drug- exposure. We seek to advance a new assay platform to effectively monitor newborns' exposure to oxycodone (oxy). Our approach will be based on two innovative approaches: extracellular vesicles (EVs) as a biomarker and iMEX (integrated magneto-electrochemical exosome) as a sensor platform. Aim 1. We will identify EV protein signatures of high-risk oxy-exposure. We will use our rodent models to emulate in-utero and postnatal oxy-exposures. EVs from brains of oxy-exposed offspring will be collected and analyzed via quantitative proteomics to identify differentially-expressed proteins. Aim 2. We will implement the second generation iMEX with significantly expand analytical capacities. We will enhance iMEX detection sensitivity by exploring a new signal amplification (nanoplasmonics); establish a unified assay to detect both transmembrane and intravesicular markers; and construct a high-throughput detector (a 96 well-plate format). This new system will be used to screen plasma EVs from oxy-exposed animals. The success of this project will generate comprehensive protein data on brain-derived EVs under oxy-exposure and critically EVs' potential as a biomarker. Furthermore, the insights gained will set the stage to further extend HiMEX technology for clinical validation. Our long-term goal is to deliver a minimally-invasive blood test for risk assessment and timely intervention for oxy-exposure.