Understanding the miRNA response to opioid withdrawal and their uses as potential biomarkers for neonatal abstinence syndrome

了解 miRNA 对阿片类药物戒断的反应及其作为新生儿戒断综合征潜在生物标志物的用途

• 批准号: 10536908
• 负责人: Rhea Elena Sullivan
• 金额: $ 3.35万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10536908
• 关键词: Admission activity; Adult; Affect; Age; Animal Model; Binding; Biological; Biological Assay; Biological Markers; Biology; Birth; Blood; Brain; Caring; Cell Differentiation process; Cell Maturation; Cell Survival; Cerebrospinal Fluid; Childhood; Clinical; Clinical Management; Clinical assessments; Collection; Complex; Cranial Nerves; Cyclic AMP-Dependent Protein Kinases; Data; Doctor of Philosophy; Dose; Early Intervention; Enrollment; Fellowship; Gene Expression Regulation; Genetic Transcription; Genetic Translation; Glial Fibrillary Acidic Protein; Hospitalization; Hospitals; Hour; Human; Hyperactivity; Impairment; In Vitro; Incidence; Infant; Knowledge; Lead; Length of Stay; Linear Regressions; Longitudinal cohort study; Measures; Mediating; Medical; Messenger RNA; MicroRNAs; Molecular; Morphine; Neonatal Abstinence Syndrome; Neuraxis; Neuroglia; Neurons; Newborn Infant; Nucleic Acids; Oligodendroglia; Opiate Addiction; Opioid; Oropharyngeal; Outcome; Physiological; Polymerase Chain Reaction; Questionnaires; RNA; Race; Reader; Risk; Saliva; Salivary; Serum; Small RNA; Source; Symptoms; System; Testing; Transcript; Transfection; United States; Untranslated RNA; Withdrawal; Work; base; cost; design; exosome; experience; fetal opioid exposure; immunocytochemistry; improved; infant outcome; mRNA Expression; maternal opioid use; nerve stem cell; neurodevelopment; neurogenesis; neuron development; novel therapeutics; opioid epidemic; opioid exposure; opioid withdrawal; potential biomarker; response; sample collection; service intervention; sex; single-cell RNA sequencing; tool

Project Summary/Abstract Neonatal abstinence syndrome (NAS) is characterized by central nervous system hyperactivity that occurs when an infant experiences withdrawal from maternal opioid use at birth. Rates of NAS in the United States have skyrocketed amidst the opioid epidemic. Assessment and clinical management of NAS rely on subjective symptom scales, which may contribute to the prolonged hospital stays and poor neuro- developmental outcomes associated with NAS. Enhancing our knowledge about the molecular factors that regulate the biologic response to opioid withdrawal in developing infants will provide an opportunity to create objective clinical tests and novel therapies for this significant medical problem. To date, there is no biologic tool to determine the necessary morphine dose in withdrawing infants, or to predict which infants will experience neurodevelopmental delays. This is partly due to the fact that prior studies have largely focused on the molecular response to opioid administration in adults, rather than the response to opioid withdrawal in infants. Serum levels of certain micro-ribonucleic acids (miRNAs) are impacted by opioid administration in adults (e.g., let-7a, miR-146a, miR-192). These short, non-coding nucleic acids also regulate neurogenesis, neuronal progenitor cell (NPC) maturation, and cell survival by repressing target messenger RNAs in the Argonaute complex. My sponsor, Dr. Steve Hicks, MD, PhD has pioneered the use of salivary miRNAs as non-invasive markers for pediatric neurodevelopmental conditions. Most salivary miRNAs are derived from exosomes, which can arise from the cranial nerves that densely innervate the oropharynx. My preliminary data shows that several “opioid-responsive” miRNAs are perturbed in the saliva of infants with NAS relative to healthy infants. Further, I have developed an in vitro system of opioid withdrawal utilizing human NPCs that displays dose- related perturbations in miR-146a along with disruptions in NPC fate. Based on these findings, the central hypothesis of my fellowship application is that morphine withdrawal alters miRNA expression in the developing brain, which impairs neuronal maturation by mRNA translation via Argonaute binding. I also hypothesize that salivary levels of exosomal miRNAs from infants with NAS will be directly related to both the maximal dose of morphine required for symptom control, and neurodevelopmental outcomes at six months. I will test these hypotheses in two specific aims. First, I will perform a longitudinal cohort study of 50 infants with NAS to determine whether salivary miRNA levels within brain-related exosomes can be measured at admission to predict the maximum morphine dose required for symptom control, and again at discharge to predict neurodevelopmental outcomes at six months (Aim 1). Second, I will transfect miRNA mimics in my in vitro NPC design of opioid withdrawal, and assess the mechanism by which miRNAs impact the response to opioid withdrawal with immunocytochemistry, single-cell RNA sequencing, and an Argonaute pull-down assay (Aim 2).

项目摘要/摘要 新生儿禁欲综合征(NAS)的特征是中枢神经系统过度活跃， 发生在婴儿出生时就戒断了母亲的阿片类药物使用。NAS在美国的比率 在阿片类药物泛滥的情况下，各州的数量激增。NAS的评估和临床管理依赖于 主观症状量表，这可能是导致住院时间延长和神经功能不佳的原因。 与NAS相关的发展成果。增强我们对分子因素的了解 调节发育中婴儿对阿片类药物戒断的生物反应将提供一个创造机会 目的针对这一重大医学问题进行临床试验和新的治疗方法。到目前为止，还没有生物工具 为了确定停药婴儿所需的吗啡剂量，或预测哪些婴儿将经历 神经发育迟缓。这在一定程度上是因为以前的研究主要集中在 成人对阿片类药物使用的分子反应，而不是婴儿对阿片类药物戒断的反应。 成人服用阿片类药物会影响血清某些微核糖核酸(MiRNAs)的水平(例如， LET-7a、miR-146a、miR-192)。这些短的、非编码的核酸还调节神经发生、神经元 鼻祖细胞(NPC)的成熟和通过抑制靶信使RNA的细胞存活 很复杂。我的赞助人史蒂夫·希克斯博士，医学博士，博士，开创了唾液miRNAs的非侵入性使用 儿童神经发育状况的标志物。大多数唾液中的miRNAs来源于外切体，而外切体 可起源于密集支配口咽部的脑神经。我的初步数据显示 与健康婴儿相比，NAS婴儿唾液中的几个“阿片反应”miRNAs受到干扰。 此外，我还开发了一种阿片类药物戒断的体外系统，该系统利用人的鼻咽癌细胞显示剂量- MiR-146a中的相关扰动以及NPC命运的中断。根据这些调查结果，中央 我的奖学金申请假设是吗啡戒断改变了发育过程中miRNA的表达 大脑，通过Argavite结合，通过mRNA翻译来损害神经元的成熟。我还假设 NAS婴儿唾液外体miRNAs水平与最大剂量 症状控制所需的吗啡，以及六个月后的神经发育结果。我要测试一下这些 两个特定目标的假设。首先，我将对50名患有NAS的婴儿进行纵向队列研究，以 确定是否可以在入院时测量脑相关外切体中的唾液miRNA水平 预测症状控制所需的最大吗啡剂量，并在出院时再次预测 6个月时的神经发育结果(目标1)。其次，我将在我的体外鼻咽癌中导入miRNA模拟物。 设计阿片类药物戒断，并评估miRNAs影响阿片类药物反应的机制 通过免疫细胞化学、单细胞RNA测序和ArgAerte下拉试验(AIM)停药 2)。