Assessment of fetal brain health via circulating exRNA carriers for opioid use disorder in pregnancy

项目摘要 我们提交此提案是为了响应PAR-20-147（细胞外RNA载体亚类， 与物质使用障碍或艾滋病毒感染有关的过程）。 阿片类药物在美国的流行对公共卫生产生了深远的影响。近几十年来， 患有新生儿戒断综合征（NAS）的新生儿出生率急剧增加。但并非 所有在妊娠期间暴露于阿片类药物的新生儿都会发生NAS或需要治疗。NAS是一个复杂的， 特异质条件，目前甚至在出生时也不可能预测哪些婴儿将需要 药物治疗或易受其最坏结果的影响。胎儿脑和CNS的生物标志物 阿片类药物暴露期间的发育对于决定哪些婴儿需要阿片类药物治疗非常有价值 为NAS和阿片类药物暴露的婴儿进行个性化的风险评估。他们还可以揭示 导致妊娠期间阿片类药物暴露最严重影响的生物学途径。 来源于胎脑和中枢神经系统（FNEV）的细胞外囊泡（EV）可以穿过神经系统， 在一些实施方案中，所述载体可以是胎儿/胎盘屏障，并且可以从母体血液中分离。它们代表了一个强大的可用工具， 提供胎儿神经发育和损伤的见解。然而，电动汽车是高度异质性的， 一组表面蛋白和囊内货物，如RNA。目前的分离和研究方法 EV缺乏必要的灵敏度和精确度来表征EV亚群。应对当前 EV分析的局限性，我们开发了一种高通量的单EV流分析仪和分选仪， 检测单个染料分子。 这种新的技术能力使我们能够精确地定义妊娠期间血浆FNEV的特征。 我们的研究可能会导致测定，定义NAS风险出生前，并带来新的见解的机制， 胎儿脑损伤的途径。

Project Summary We are submitting this proposal in response to PAR-20-147 (Extracellular RNA carrier subclasses in processes relevant to Substance Use Disorders or HIV infection). The opioid epidemic in the United States has far reaching consequences for public health. In recent decades, the rate of newborns born with neonatal abstinence syndrome (NAS) has increased dramatically. However, not all neonates exposed to opioids during gestation develop NAS or require treatment. NAS is a complex and idiosyncratic condition, and it is currently not possible to predict even at birth which infants will require pharmacotherapy or be susceptible to its worst outcomes. Accessible biomarkers for fetal brain and CNS development during opioid exposure would be highly valuable for deciding which infants will need opioid therapy for NAS and making personalized risk assessments for opioid exposed infants. They could also shed light on the biological pathways that lead to the worst effects of opioid exposure during gestation. Extracellular vesicles (EVs) derived from the Fetal brain and central Nervous system (FNEV) can cross the fetal/placental barrier and can be isolated from maternal blood. They represent a powerful accessible tool to provide insight on fetal neurodevelopment and damage. However, EVs are highly heterogeneous, with a diverse set of surface proteins and intra-vesicular cargo, such as RNA. Current approaches to the isolation and study of EVs lack the necessary sensitivity and precision to characterize EV subpopulations. To address the current limitations of EV analyses, we developed a high-throughput single-EV flow analyzer and sorter capable of detecting single dye molecules. This new technical capability allows us to exquisitely define the profiles of plasma FNEV during pregnancy. Our studies could lead to assays that define NAS risk pre-birth and bring new insights into the mechanistic pathways of fetal brain injury.