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 降低戒断后复发率是遏制阿片类药物流行的一个关键挑战。一 触发复发的常见因素之一是重新接触与药物有关的线索。在大鼠中，线索诱导 在停止长期使用羟考酮的前几周，羟考酮的寻求逐渐加剧（潜伏） 羟考酮自我管理，并保持高至少一个月的禁欲。我们最近发表的工作 证明了眶额皮质（OFC）在这种孵化中的关键作用。然而， 这种孵化的基础OFC在很大程度上是未知的。具体来说，我们观察到， OFC中与羟考酮寻求相关的神经元激活可能是时间依赖性分子机制的结果。 戒断期间眶额皮层的适应性变化新出现的证据表明泛素蛋白酶体系统 (UPS)和线粒体调节成瘾相关的可塑性。将这两条工作线联系起来，我们发现， 线粒体E3泛素连接酶March 5的mRNA表达在OFC中显著降低， 与生理盐水对照相比，禁欲15天。March 5的一个关键作用是抑制线粒体分裂， 降解线粒体分裂介质，如动力蛋白相关蛋白1（Drp 1）。根据初步的 工作，在这里，我们建议检查3月5日的作用和相关的线粒体动力学在孵育 对羟考酮的渴望在目的1中，我们将使用生物化学方法来检测March 5和 它的下游靶点是OFC中的Drp 1，并使用病毒介导的基因操作来确定March 5是否在OFC中。 OFC在羟考酮渴求的孵化中起着因果作用。在目标2中，我们将使用病毒载体标记 OFC神经元的线粒体，并检查孵育过程中OFC神经元线粒体的形态 对羟考酮的渴望总的来说，这项提案将启动一个新的研究线，重点是线粒体E3 OFC中的泛素连接酶在药物寻求复发中的作用从临床的角度来看，这项工作将提供目标， * 制定药物干预措施，以减少对毒品的渴望并促进戒断。