Cell type transcriptional mechanisms of polysubstance choice

多物质选择的细胞类型转录机制

• 批准号: 10740057
• 负责人: Matthew L Banks
• 金额: $ 44.31万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 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/10740057
• 关键词: Area; Behavior; Behavioral; Behavioral Model; Biological Assay; Brain; Brain region; Cessation of life; Choice Behavior; Classification; Clinical; Consumption; Data; Data Set; Decision Making; Drug Addiction; Drug usage; Female; Fentanyl; Food; Gene Expression Profile; Genetic Transcription; Goals; Human; Medial; Methamphetamine; Modeling; Molecular; Neuroglia; Nucleus Accumbens; Opioid; Pathogenesis; Pharmaceutical Preparations; Phase; Prefrontal Cortex; Procedures; Process; Public Health; Publishing; Rattus; Recording of previous events; Recovery; Research; Research Priority; Rewards; Risk; Saline; Self Administration; Severities; Stimulant; Testing; Tissues; Transcript; Withdrawal; Work; brain cell; brain reward regions; cell type; comorbidity; drug reinforcement; drug use behavior; experience; fentanyl self-administration; fentanyl use; improved; male; methamphetamine use; non-drug; novel; opioid misuse; opioid use; pharmacologic; polysubstance use; pre-clinical; pre-clinical research; reinforcer; response; single nucleus RNA-sequencing; stimulant misuse; stimulant use disorder; substance use; synergism; transcriptome sequencing

PROJECT SUMMARY In this new R01 application that is directly responsive to RFA-DA-23-015, our research team proposes to study the cell type specific molecular mechanisms regulated by fentanyl and methamphetamine polysubstance use at both the initiation and withdrawal stages of the substance use trajectory. We propose to overcome current limitations in polysubstance research by utilizing preclinical assays of drug-vs.-food choice procedures in male and female rats, which more fully capture the severity of polysubstance use seen in humans by modeling the behavioral misallocation and decision making between concurrently available addictive drugs and alternative non-drug reinforcers. We will combine these enhanced behavioral models with single nuclei RNA sequencing (snRNAseq) of the medial prefrontal cortex (PFC) and nucleus accumbens (NAc), key brain regions implicated in drug reinforcement and drug-taking, to capture and characterize the exact drug-induced molecular adaptations that occur in specific cell types, including non-neuronal cells. We will directly test the hypothesis that the synergistic action of combined fentanyl and methamphetamine use produces enhanced drug use behaviors and brain molecular adaptations that are distinct from what is achieved by either fentanyl or methamphetamine use alone; that this polysubstance synergy involves unique transcriptional adaptations by brain region, accumulates as a function of drug experience, and contributes to the behavioral misallocation towards drug use over more beneficial rewarding activities that is the hallmark of drug addiction. We will test this overarching hypothesis in two Aims. In Aim 1, we will uncover the impact of fentanyl/methamphetamine polysubstance use during the withdrawal phase of the substance use trajectory. We will use drug-vs.-food self- administration choice procedures for saline, fentanyl alone, methamphetamine alone, and fentanyl/methamphetamine combinations to uncover how an extended history of polysubstance use synergizes to increase somatic withdrawal effects and drug taking behavior while experiencing withdrawal. We will then perform snRNAseq in the PFC and NAc to interrogate the brain cell type specific transcriptional adaptations in these rats. In Aim 2, we will similarly perform drug-vs.-food choice procedures and snRNAseq of these brain regions to explore the emergence of behavioral and transcriptional adaptations at the of initiation of drug use experience. We will go on to compare our snRNAseq data from Aims 1 and 2 to understand how the fentanyl/methamphetamine polysubstance cell type transcriptional profile changes over the substance use trajectory. This project will reveal how fentanyl and methamphetamine synergize to produce maladaptive drug choice behaviors and brain cell type specific transcriptional responses at distinct stages of the substance use trajectory that are common barriers to recovery. Results gained from this project will inform the discovery of novel and more efficacious pharmacological agents to treat the core decision making process that is uniquely disrupted by polysubstance use.

项目摘要 在这个直接响应RFA-DA-23-015的新R 01应用中，我们的研究团队建议研究 芬太尼和甲基苯丙胺多物质使用调节的细胞类型特异性分子机制 在物质使用轨迹的开始和停止阶段。我们建议克服目前 通过利用药物-vs.-的临床前测定进行多物质研究的局限性男性食物选择程序 和雌性大鼠，它们通过模拟 同时可用的成瘾性药物和替代药物之间的行为错误分配和决策 非药物制剂。我们将联合收割机将这些增强的行为模型与单核RNA测序相结合 内侧前额叶皮层（PFC）和脑桥核（NAc）的snRNAseq（snRNAseq）， 在药物强化和药物服用中，捕获和表征精确的药物诱导分子， 适应发生在特定的细胞类型，包括非神经元细胞。我们将直接检验假设 芬太尼和甲基苯丙胺联合使用的协同作用会增加药物使用， 行为和大脑分子适应，这与芬太尼或 甲基苯丙胺单独使用;这种多物质协同作用涉及独特的转录适应， 大脑区域，积累作为药物经验的函数，并有助于行为的错误分配 而不是更有益的奖励活动，这是吸毒成瘾的标志。我们将测试 这一总体假设有两个目标。在目标1中，我们将揭示芬太尼/甲基苯丙胺的影响 在物质使用轨迹的戒断阶段使用多种物质。我们将使用药物-vs。食物自 盐水、单独芬太尼、单独甲基苯丙胺的给药选择程序，以及 芬太尼/甲基苯丙胺组合，以揭示多种物质使用的长期历史如何协同作用 在经历戒断时增加躯体戒断效应和吸毒行为。然后我们将 在PFC和NAc中进行snRNAseq，以询问脑细胞类型特异性转录适应， 这些老鼠在目标2中，我们将类似地执行药物vs.这些大脑的食物选择程序和snRNAseq 区域，以探索在开始使用药物时出现的行为和转录适应 体验.我们将继续比较目标1和目标2的snRNAseq数据，以了解 芬太尼/甲基苯丙胺多物质细胞类型转录谱随物质使用的变化 弹道该项目将揭示芬太尼和甲基苯丙胺如何协同产生适应不良的药物 在物质使用的不同阶段的选择行为和脑细胞类型特异性转录反应 这是复苏的常见障碍。从这个项目中获得的结果将有助于发现 新的和更有效的药物治疗的核心决策过程， 被多种物质的使用所破坏。