Identifying multi-omic signatures of opioid use and relapse

识别阿片类药物使用和复发的多组学特征

• 批准号: 10724297
• 负责人: Megan Elizabeth Fox
• 金额: $ 49.8万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10724297
• 关键词: Amygdaloid structure; Animals; Automobile Driving; Award; Behavior; Brain; Calcium; Cementation; Clustered Regularly Interspaced Short Palindromic Repeats; DNA Methylation; Data; Development; Disease; Drug usage; Epigenetic Process; Financial cost; Genes; Genetic Transcription; Genetic study; Health Care Costs; Individual; Intake; Methylation; Molecular; Neurons; Nucleus Accumbens; Opioid; Pharmaceutical Preparations; Physiology; Public Health; Recovery; Relapse; Research; Resolution; Rewards; Self Administration; Substance Use Disorder; Synaptic plasticity; Testing; United States; Ventral Tegmental Area; demethylation; differential expression; fentanyl self-administration; gene network; medication-assisted treatment; mouse model; multiple omics; neuromechanism; new therapeutic target; novel therapeutic intervention; novel therapeutics; opioid epidemic; opioid exposure; opioid overdose; opioid use; opioid use disorder; opioid user; prolonged abstinence; promoter; relapse prevention; support network

Project Summary Opioid use disorder is a life-long burden for many individuals, imposing high personal, financial, and health costs. Even after prolonged abstinence, many individuals in recovery will go on to relapse, including those that received medication-assisted treatments. Repeated opioid exposure usurps normal reward circuit function by producing long-lasting molecular changes that alter physiology and support continued drug use. These cellular adaptations have been implicated in sustained relapse vulnerability, but we lack a clear understanding of what drives their persistence. There is a further lack of information on the precise molecular adaptations underlying altered circuit function, and in which specific circuits they act to promote relapse. Understanding this “who, what, when, and where,” will be key to identifying new therapeutic targets. Here, we will answer these questions using a multi- level approach that allows us to sequence, manipulate, and record from neurons in specific circuits in the context of opioid self-administration and relapse. Our preliminary data show that both genetically-distinct and genetically-identical neuron subtypes in the ventral tegmental area (VTA) undergo differential molecular adaptations after fentanyl self-administration, which we hypothesize arises from activity-dependent transcriptional changes in specific circuits. We further hypothesize the transcriptional changes are sustained by methylation and demethylation at the gene promoters. We will first record calcium activity in VTA neurons that project to either the nucleus accumbens (NAc) or amygdala (AMY)— projections known to be important for drug intake and relapse, respectively. Then, in the same neurons from the same animals, we will identify which gene networks are transcriptionally changed after self-administration and persist until relapse testing. Next, we will identify the DNA methylation marks driving sustained differential expression, with an emphasis on genes important for synaptic plasticity. Next, we will use CRISPR/dCas9 fusion constructs to manipulate methylation states at our identified loci in specific circuits. This proposal will allow us to define the specific VTA circuits that support opioid intake and relapse, which gene networks support activity of these circuits, and how DNA methylation cements the transcriptional landscape to alter behavior. This award will allow research into neural mechanisms of opioid use disorder with unprecedented resolution, and has the potential to transform how we approach studying the genetics of substance use disorders. Together, this critical information will help inform new treatment strategies to prevent relapse.

项目摘要 阿片类药物使用障碍对许多人来说是一个终身的负担，造成高昂的个人、经济和健康成本。 即使在长期禁欲之后，许多正在康复的人会继续复发，包括那些接受了 药物辅助治疗。反复接触阿片类药物通过产生 长期的分子变化，改变生理并支持持续的药物使用。这些细胞适应 与持续的复发脆弱性有关，但我们缺乏对他们的驱动因素的清楚了解 坚持不懈。更缺乏关于改变回路背后的精确分子适应的信息。 功能，以及它们在哪些特定的回路中起促进复发的作用。理解这个“谁、什么、何时、以及 在哪里，“将是确定新的治疗靶点的关键。在这里，我们将使用多个- 电平方法，允许我们对上下文中特定电路中的神经元进行排序、操作和记录 阿片类药物自我给药和复发的可能性。 我们的初步数据显示，腹侧的神经元亚型既有遗传上不同的，也有遗传上相同的 芬太尼自我给药后，被盖区(VTA)经历了不同的分子适应，我们 假说源于特定回路中依赖活性的转录变化。我们进一步假设 基因启动子上的甲基化和去甲基化维持了转录变化。我们将首先 记录VTA神经元投射到伏隔核(NAC)或杏仁核(AMY)的钙活动。 已知的预测分别对药物摄取和复发非常重要。然后，在相同的神经元中，来自 同样的动物，我们将识别哪些基因网络在自我给药和 坚持到复发测试为止。接下来，我们将确定驱动持续差异的DNA甲基化标记 表达，重点是对突触可塑性重要的基因。接下来，我们将使用CRISPR/dCas9融合 构建来操纵我们在特定回路中识别的基因座上的甲基化状态。这项提议将使我们能够 确定支持阿片类药物摄取和复发的特定VTA回路，哪些基因网络支持阿片类药物的活性 这些回路，以及DNA甲基化如何巩固转录格局，以改变行为。这一奖项将 允许以前所未有的分辨率研究阿片类药物使用障碍的神经机制，并具有 有可能改变我们研究物质使用障碍遗传学的方式。总而言之，这一点至关重要 信息将有助于为新的治疗策略提供信息，以防止复发。