March5 and Associated Mitochondrial Dynamics in Incubation of Oxycodone Craving

March5 和相关线粒体动力学在羟考酮渴望孵化中的作用

• 批准号: 10724668
• 负责人: Xuan Anna Li
• 金额: $ 22.47万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10724668
• 关键词: Abstinence; Anatomy; Behavior; Biochemical; Cell physiology; Clinical; Cues; Data; Drug Addiction; Drug usage; Dynamin; Exploratory/Developmental Grant; Exposure to; Extinction; Frequencies; Incubated; Intervention; Label; Lead; Length; Link; Literature; Mediating; Mediator; Mitochondria; Modeling; Molecular; Molecular Target; Morphine; Morphology; Mus; Neurons; Nucleus Accumbens; Opiate Addiction; Opioid; Oxycodone; Pharmaceutical Preparations; Play; Proteins; Publishing; Rattus; Regulation; Relapse; Research; Rewards; Role; Saline; Self Administration; System; Testing; Time; Ubiquitin; Ubiquitination; Viral; Viral Vector; Work; activity marker; addiction; cocaine relapse; cocaine seeking; craving; drug abstinence; drug craving; genetic manipulation; innovation; mRNA Expression; male; mitochondrial dysfunction; multicatalytic endopeptidase complex; neuromechanism; opioid epidemic; pharmacologic; prescription opioid abuse; prolonged abstinence; protein degradation; protein expression; psychostimulant; relapse prevention; therapeutic target; ubiquitin-protein ligase

Project summary/abstract Reducing rates of relapse after abstinence is a key challenge for curbing the ongoing opioid epidemic. One of the common factors for triggering relapse is re-exposure to drug-associated cues. In rats, cue-induced oxycodone seeking progressively intensifies (incubates) over the first weeks of abstinence from extended access oxycodone self-administration, and remains high for at least a month of abstinence. Our recently published work demonstrated a critical role of orbitofrontal cortex (OFC) in this incubation. However, molecular mechanisms in OFC underlying this incubation are largely unknown. Specifically, our observation of a progressive increase of neuronal activation in OFC associated with oxycodone seeking could be the result of time-dependent molecular adaptations in OFC during abstinence. Emerging evidence has implicated both the ubiquitin proteasome system (UPS) and mitochondrial regulation in addiction-related plasticity. Linking these two lines of work, we found that mRNA expression of March5, a mitochondrial E3 ubiquitin ligase, was significantly decreased in OFC on abstinence day 15 compared with saline control. A key role of March5 is to suppress mitochondrial fission, by degrading mitochondrial fission mediators such as dynamin-related protein 1(Drp1). Based on this preliminary work, here we propose to examine the role of March5 and associated mitochondria dynamics in incubation of oxycodone craving. In Aim 1, we will use biochemical approaches to examine protein expression of March5 and its downstream target Drp1 in the OFC, and use viral-mediated gene manipulation to determine if March5 in the OFC plays a causal role in the incubation of oxycodone craving. In Aim 2, we will use viral vectors to label mitochondria in OFC neurons and examine the morphology of mitochondria in OFC neurons during incubation of oxycodone craving. Overall, this proposal will initiate a new line of research focusing on the mitochondrial E3 ubiquitin ligase in OFC in relapse to drug seeking. From a clinical perspective, this work will provide targets for developing pharmacological interventions to decrease drug craving and promote abstinence.

项目摘要/摘要 减少戒毒后的复发率是遏制持续的阿片类药物流行的关键挑战。一 引发复发的常见因素之一是再次接触与药物有关的线索。在大鼠中，线索诱导 在戒烟的头几周内，羟考酮的寻求逐渐加强(孵化)，不再长期使用 羟考酮自我给药，并在至少一个月的禁欲中保持高水平。我们最近出版的作品 证实了眶前皮质(OFC)在这一孵化过程中的关键作用。然而，分子机制在 这种孵化背后的OFC在很大程度上是未知的。具体地说，我们观察到的渐进增长 OFC中与羟考酮寻找相关的神经元激活可能是时间依赖性分子的结果 禁欲期间OFC的适应。新出现的证据表明，泛素蛋白酶体系统 (UPS)和成瘾相关可塑性中的线粒体调节。把这两条线联系起来，我们发现 OFC中线粒体E3泛素连接酶March5的mRNA表达显著降低。 戒断第15天与生理盐水对照组比较。March5的一个关键作用是抑制线粒体分裂，通过 降解线粒体分裂介体，如动力蛋白相关蛋白1(Drp1)。根据这一初步情况 在这项工作中，我们建议研究March5及其相关的线粒体动力学在胚胎发育过程中的作用 对羟考酮的渴望。在目标1中，我们将使用生化方法来检测March5和March5的蛋白表达 它的下游靶向OFC中的Drp1，并使用病毒介导的基因操作来确定March5在OFC中是否 OFC在羟考酮渴求的孵化中起因果作用。在目标2中，我们将使用病毒载体标记 OFC神经元线粒体及孵育过程中线粒体形态的观察 对羟考酮的渴望。总体而言，这项提议将开启一条新的研究路线，专注于线粒体E3 泛素连接酶在复发性药物依赖中的作用。从临床的角度来看，这项工作将为 开发药物干预措施，减少药物渴求，促进戒毒。