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).

项目概要/摘要 新生儿戒断综合征（NAS）的特点是中枢神经系统过度活跃， 当婴儿在出生时停止使用母亲的阿片类药物时就会发生这种情况。美国 NAS 费率 由于阿片类药物的流行，各州的数量猛增。 NAS的评估和临床管理依赖于 主观症状量表，这可能导致住院时间延长和神经功能不佳 与 NAS 相关的发育结果。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机会 针对这一重大医学问题的客观临床测试和新疗法。迄今为止，还没有任何生物工具 确定戒断婴儿所需的吗啡剂量，或预测哪些婴儿会经历 神经发育迟缓。部分原因是之前的研究主要集中在 成人对阿片类药物给药的分子反应，而不是婴儿对阿片类药物戒断的反应。 某些微核糖核酸 (miRNA) 的血清水平受到成人阿片类药物给药的影响（例如， let-7a、miR-146a、miR-192）。这些短的非编码核酸还调节神经发生、神经元 通过抑制 Argonaute 中的靶信使 RNA 来促进祖细胞 (NPC) 成熟和细胞存活 复杂的。我的资助者 Steve Hicks 博士率先使用唾液 miRNA 作为非侵入性治疗 儿科神经发育状况的标志物。大多数唾液 miRNA 来源于外泌体， 可由密集支配口咽的脑神经产生。我的初步数据表明 与健康婴儿相比，NAS 婴儿唾液中的几种“阿片类药物反应性”miRNA 受到干扰。 此外，我还开发了一种利用人类 NPC 的阿片类药物戒断体外系统，该系统显示剂量 miR-146a 的相关扰动以及 NPC 命运的破坏。根据这些发现，中央 我的奖学金申请假设是吗啡戒断会改变发育中的 miRNA 表达 大脑，通过 Argonaute 结合进行 mRNA 翻译，从而损害神经元成熟。我还假设 患有 NAS 的婴儿的外泌体 miRNA 的唾液水平将与最大剂量直接相关 吗啡用于控制症状和六个月时的神经发育结果。我会测试这些 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大脑相关外泌体中的唾液 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 六个月时的神经发育结果（目标 1）。其次，我将在体外 NPC 中转染 miRNA 模拟物 设计阿片类药物戒断，并评估 miRNA 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 Argonaute Pull-down 测定（Aim 2）。