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

基本信息

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
项目类别：
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

项目摘要

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终生烧伤，施加了高昂的个人，财务和健康成本。即使在节制长期之后，许多康复中的人也会继续缓解，包括那些接受的人药物辅助治疗。反复的阿片类药物暴露通过产生正常奖励电路功能持久的分子变化改变了生理和支持继续使用药物。这些细胞适应在持续的继电器脆弱性中隐含了持久性。进一步缺乏有关改变电路的确切分子适应的信息功能，以及它们在哪些特定电路中起作用来促进救济。理解这个“谁，什么，何时和在哪里，“将是识别新的治疗靶标的关键。在这里，我们将使用多种多数问题回答这些问题使我们能够在特定圆圈中对神经元进行测序，操纵和记录的水平方法阿片类型的自我管理和继电器。我们的初步数据表明，腹侧的遗传差异和遗传性神经元亚型芬太尼自我给药后的换段区域（VTA）经历差异分子适应假设是由特定电路的活动依赖性转录变化引起的。我们进一步假设转录变化是通过基因启动子的甲基化和脱甲基化来维持的。我们将首先在VTA神经元中记录钙活性，这些钙活性将投射到伏隔核（NAC）或杏仁核（Amy） - 已知对药物摄入和救济至关重要的预测。然后，在同一神经元中同一动物，我们将确定自我管理后哪些基因网络在转录上发生了变化和一直持续到继电器测试。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融合在我们确定的特定电路中，在我们确定的局部操纵甲基化状态的构造。该建议将使我们能够定义支持阿片类药物摄入和继电器的特定VTA电路，基因网络支持的活动这些电路以及DNA甲基化如何巩固转录局势以改变行为。这个奖项将允许研究阿片类药物使用障碍的神经机制，并具有前所未有的分辨率，并具有改变我们如何研究物质使用障碍的遗传学的潜力。在一起，这一点信息将有助于为防止退休的新治疗策略提供信息。