CELL-TYPE SPECIFIC ROLE OF CIRCADIAN-DEPENDENT TRANSCRIPTION IN FENTANYL-INDUCED SYNAPTIC AND BEHAVIORAL PLASTICITY

昼夜节律依赖性转录在芬太尼诱导的突触和行为可塑性中的细胞类型特异性作用

• 批准号: 10830682
• 负责人: Ryan W Logan
• 金额: $ 6.02万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-23 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10830682
• 关键词: CELL; TYPE; SPECIFIC; ROLE; CIRCADIAN

PROJECT ABSTRACT Proposal is designed in response to NHLBI/NIDA RFA-HL-19-028 (Sleep and Circadian-Dependent Mechanisms Contributing to Opioid Use Disorder (OUD) and Response to Medication Assisted Treatment (MAT)) as part of the HEAL Initiative. Annual costs attributed to the opioid crisis in the United States have shattered previous estimates. Synthetic opioids like fentanyl have surpassed prescription opioids as the leading cause of death from drug overdose and illicit fentanyl use continues to ride in regions with already high base rates of OUD. Among the most common symptoms experienced by individuals suffering with OUD are severe and persistent sleep and circadian disruptions. These sleep and circadian abnormalities are speculated to foster opioid craving and relapse, although direct evidence is limited. We lack a basic understanding of the mechanisms contributing to opioid dependence and their relationship to sleep and circadian systems. We identified a circadian-dependent mechanism which modulates fentanyl reward-related behaviors potentially via cell-type specific action in a key brain region linked to OUD—the nucleus accumbens (NAc). A majority of the neurons in the NAc are GABAergic medium spiny neurons which predominantly express either dopamine 1 or 2 receptors (D1R-MSNs or D2R-MSNs). These MSN subpopulations exert opposing and sometimes complementary actions on reward-related behaviors, whereby activation of D1R-MSNs drives drug seeking and relapse, while D2R-MSNs attenuate these behaviors. D1R-MSNs are also involved in the regulation of sleep-wake cycles. Our preliminary work suggests the diurnal excitability of NAc D1R-MSNs and their response to fentanyl is directly modulated by the circadian transcription factor NPAS2. We also show NPAS2 impacts sleep-wake cycles, potentially via action in NAc. Findings from human genetics studies indicate variants of NPAS2 are associated with alterations in the diurnal variation of anhedonia and motivation, suggesting links between NPAS2 and the circadian modulation of reward and motivation. We identified a novel role for NPAS2 to regulate fentanyl reward via activity in D1-MSNs of the NAc. We will use a combination of behavioral (i.e., self-administration and sleep polysomnography), slice electrophysiology, and molecular (i.e., cell-type specific RNA-seq) approaches to: Investigate the role of NPAS2 in D1R-MSNs to modulate the behavioral responses to fentanyl (Aim 1); Assess the impact of fentanyl on synaptic plasticity at D1R-MSNs and investigate whether NPAS2 mediates the potentiation of excitatory synapses at specific diurnal phases (Aim 2); Elucidate the cell-type specific NPAS2-dependent transcriptional mechanisms of fentanyl-seeking and relapse behaviors (Aim 3); and, Investigate whether NPAS2 rescue and buprenorphine MAT improve fentanyl-induced sleep disturbances (Aim 4). Our proposal will define the role for circadian-dependent transcriptional mechanisms in key reward circuits of opioid reward and uncover the therapeutic potential of NPAS2 for opioid dependence and relapse. Our studies will provide novel insights into the mechanisms at the intersection of sleep, circadian rhythms, and opioids.

项目摘要 该提案旨在响应NHLBI/NIDA RFA-HL-19-028（睡眠和昼夜节律依赖机制 阿片类药物使用障碍（OUD）和对药物辅助治疗（MAT）的反应）的一部分， 治愈计划。美国阿片类药物危机造成的年度成本打破了先前的估计。 芬太尼等合成阿片类药物已超过处方阿片类药物，成为药物死亡的主要原因。 过量和非法芬太尼的使用继续在OUD基础率已经很高的地区蔓延。的最 患有OUD的个体经历的常见症状是严重和持续的睡眠和昼夜节律 干扰据推测，这些睡眠和昼夜节律异常会促进阿片类药物的渴望和复发，尽管 直接证据有限。我们缺乏对阿片类药物依赖机制的基本了解 以及它们与睡眠和昼夜节律系统的关系。我们确定了一种昼夜节律依赖机制， 可能通过细胞类型特异性作用调节芬太尼奖励相关的行为， OUD--丘脑核（NAc）。NAc中的大多数神经元是GABA能中型棘神经元 其主要表达多巴胺1或2受体（D1 R-MSN或D2 R-MSN）。这些MSN 亚群体对奖励相关行为施加相反的，有时是互补的行为， D1 R-MSN的激活驱动药物寻求和复发，而D2 R-MSN减弱这些行为。D1R-MSN 也参与调节睡眠-觉醒周期。我们的初步工作表明， NAc D1 R-MSN及其对芬太尼的反应直接受昼夜节律转录因子NPAS 2调节。 我们还表明NPAS 2可能通过NAc中的作用影响睡眠-觉醒周期。人类遗传学的发现 研究表明NPAS 2的变体与快感缺乏的昼夜变化的改变有关， 动机，这表明NPAS 2与奖励和动机的昼夜节律调节之间的联系。我们确定 NPAS 2通过在NAc的D1-MSNs中的活性调节芬太尼奖励的新作用。我们将使用一个组合 行为（即，自我给药和睡眠多导睡眠描记术），切片电生理学，和分子（即， 细胞类型特异性RNA-seq）方法来：研究NPAS 2在D1 R-MSN中调节行为细胞的作用。 对芬太尼的反应（目的1）;评估芬太尼对D1 R-MSNs突触可塑性的影响，并研究 NPAS 2是否在特定的昼夜时相介导兴奋性突触的增强（Aim 2）; 芬太尼寻求和复发行为的细胞类型特异性NPAS 2依赖性转录机制（目的 3）;以及，调查NPAS 2补救和丁丙诺啡MAT是否改善芬太尼诱导的睡眠障碍 (Aim 4）。我们的建议将定义昼夜节律依赖的转录机制在关键奖励回路中的作用 阿片类药物的奖励，并揭示NPAS 2对阿片类药物依赖和复发的治疗潜力。我们的研究 将为睡眠、昼夜节律和阿片类药物的交叉机制提供新的见解。