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

项目摘要 我们提交这项建议是为了响应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， 出生时患有新生儿禁欲综合征(NAS)的新生儿比例大幅上升。然而，不是 所有在妊娠期间接触阿片类药物的新生儿都会患上NAS或需要治疗。NAS是一个复杂的 特殊情况，目前甚至在出生时也无法预测婴儿将需要哪些 药物治疗或易受其最坏后果的影响。胎脑和中枢神经系统可获得的生物标志物 在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和对接触阿片类药物的婴儿进行个性化风险评估。他们还可以揭示 导致妊娠期阿片类药物暴露最严重影响的生物途径。 来源于胎儿大脑和中枢神经系统(FNEV)的细胞外小泡(EVS)可以穿过 胎儿/胎盘屏障，可以从母体血液中分离出来。它们代表了一种强大的可访问工具 提供有关胎儿神经发育和损伤的见解。然而，电动汽车是高度异质的，具有多样化的 一组表面蛋白和囊泡内的货物，如RNA。目前分离和研究的方法 EV缺乏必要的敏感度和精确度来表征EV亚群。解决当前的问题 针对电动汽车分析的局限性，我们开发了一种高通量的单电动汽车流量分析仪和分选器，能够 检测单个染料分子。 这一新的技术能力使我们能够精确地定义怀孕期间血浆FNEV的分布。 我们的研究可能会导致对NAS风险的分析，并为NAS的发病机制带来新的见解 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.