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.

项目概要/摘要 降低戒断后的复发率是遏制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重新暴露于与药物相关的线索。在大鼠中，提示诱导 在戒断的最初几周内，对羟考酮的寻求逐渐加强（潜伏） 羟考酮自我给药，并在至少一个月的禁欲期间保持高水平。我们最近发表的作品 证明了眶额皮质（OFC）在此孵化中的关键作用。然而，分子机制 这种孵化背后的 OFC 很大程度上是未知的。具体来说，我们观察到逐渐增加 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机制的结果 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。将这两条工作联系起来，我们发现 March5（一种线粒体 E3 泛素连接酶）的 mRNA 表达在 OFC 中显着降低 禁欲第 15 天与生理盐水对照相比。 March5 的一个关键作用是抑制线粒体裂变，通过 降解线粒体裂变介质，例如动力相关蛋白 1 (Drp1)。在此基础上初步 工作，在这里我们建议检查 March5 和相关线粒体动力学在孵化中的作用 羟考酮渴望。在目标 1 中，我们将使用生化方法来检查 March5 和 其下游目标 OFC 中的 Drp1，并使用病毒介导的基因操作来确定 March5 是否在 OFC 在羟考酮渴望的产生中起着因果作用。在目标 2 中，我们将使用病毒载体来标记 OFC 神经元中的线粒体并检查孵化过程中 OFC 神经元中线粒体的形态 对羟考酮的渴望。总体而言，该提案将启动一条新的研究路线，重点关注线粒体 E3 OFC 中的泛素连接酶在药物寻求复发中的作用。从临床角度来看，这项工作将为 制定药物干预措施以减少对药物的渴望并促进戒毒。