Functional Epigenetic Profiling of Anti-Relapse Cannabidiol

抗复发大麻二酚的功能表观遗传学分析

• 批准号: 9317927
• 负责人: Nobuyoshi Suto
• 金额: $ 28.88万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9317927
• 关键词: Aftercare; Anatomy; Animal Model; Biological Availability; Brain; Cannabidiol; Cannabis sativa plant; Chronic; Clinical; Cocaine; Code; Cues; DNA; DNA Methylation; Development; Disease; Drug Addiction; Epigenetic Process; Fluorescence-Activated Cell Sorting; Gene Activation; Gene Expression; Gene Expression Profiling; Genes; Genetic Transcription; Goals; Histones; Hybrids; Knowledge; Laboratory Animals; Link; Mediating; Medical; Messenger RNA; Methodology; Methods; Modification; Molecular; Neuroglia; Neurons; Nucleus Accumbens; Opioid; Pharmaceutical Preparations; Poly A; Post-Translational Protein Processing; Proteins; Rattus; Regulator Genes; Relapse; Reporting; Research; Risk Factors; Route; Sampling; Site; Stress; Testing; Tetanus Helper Peptide; Therapeutic; Tissues; Transgenic Organisms; Translations; Vial device; alcohol seeking behavior; base; cell type; chromatin immunoprecipitation; chromatin remodeling; clinically relevant; cocaine relapse; cue reactivity; drug craving; drug modification; drug of abuse; drug relapse; epigenetic profiling; epigenome; gene repression; genome-wide; improved; interdisciplinary approach; mRNA Expression; methylation biomarker; molecular marker; neurobiological mechanism; next generation sequencing; novel; oligo (dT); relating to nervous system; response; transcriptome; transdermal cannabidiol; treatment duration

Drug addiction is a chronic relapsing disorder of compulsive drug craving, seeking and use. We have identified a substantial therapeutic potential of cannabidiol (CBD) – a non-psychoactive compound naturally occurring in the cannabis sativa plant – against drug relapse. A 7-day treatment of CBD via a clinically relevant transdermal route reduced both cue- and stress-triggered reinstatement of cocaine as well as alcohol seeking in rats – well- established animal models of drug relapse. Taken together with relevant reports on opioids, CBD appears to be effective against two major modes of relapse-promotion across three major classes of drugs of abuse. While the underlying neurobiological mechanism is still unclear, CBD's anti-relapse action (still evident at 138 days post-treatment) greatly outlasted its bioavailability (<18 days). It is thus likely that molecular alterations – via long-lasting epigenetic modifications – underlie the prolonged anti-relapse action of CBD. Previous studies have identified CBD-triggered molecular alterations in samples derived from homogenized tissues of the nucleus accumbens (NAc) – a brain site well-implicated in drug relapse. Available evidence however indicates that cue-triggered responses – such as drug craving and seeking – are mediated by `cue-reactive (activated)' rather than `non-reactive' neurons in NAc (and other sites). Given that only a small subset (3-10%) of neurons in NAc (and other sites) are reactive to drug cues, molecular alterations identified in samples derived from homogenized tissues – that contain both cue-reactive and non-reactive neurons as well as non-neural glia – may not be functionally relevant to CBD's anti-relapse action. In contrast, long-lasting epigenetic modifications unique to neurons in which drug cue-reactivity is modulated by CBD likely mediate the prolonged anti-relapse action of CBD. Based on the scientific premise outlined above, this R21 project will test the hypothesis that “CBD exerts its prolonged anti-relapse action via epigenetic modification of drug cue-reactive neurons”. The project will take a multidisciplinary approach incorporating (1) a well-established animal model of cocaine relapse, (2) a novel strain of transgenic rats for activity-dependent neural tagging, (3) cell type-specific molecular sampling via fluorescence activated cell sorting, and (4) next-generation sequencing for comprehensive profiling of epigenetic and transcriptional markers. The primary goal is to determine the long- lasting effects of CBD on gene expression – that are both cell type-specific and activity-dependent (on drug cue-reactivity) – at the level of epigenome and protein-coding transcriptome, and thereby help elucidate the molecular mechanisms functionally linked to CBD's prolonged anti-relapse action. Another primary goal is to establish the proof of concept for the functional epigenetic profiling of CBD as a potential anti-relapse medication – a novel method that can be applied to investigate the therapeutic action of other medications. The knowledge gained through such studies then can be utilized to improve the efficacy of medical treatments via precise identification of on- and off-target effects.

药物成瘾是一种慢性复发性疾病，表现为强迫性的药物渴望、寻求和使用。我们已经确定 大麻二酚 (CBD) 的巨大治疗潜力——一种天然存在于体内的非精神活性化合物 大麻植物——防止药物复发。通过临床相关的透皮疗法进行为期 7 天的 CBD 治疗 该路线减少了老鼠的提示和压力触发的可卡因恢复以及酒精寻求——嗯—— 建立药物复发动物模型。结合阿片类药物的相关报告，CBD 似乎 有效对抗三种主要滥用药物类别的两种主要复发促进模式。 虽然潜在的神经生物学机制仍不清楚，但 CBD 的抗复发作用（在 138 治疗后天）大大超过了其生物利用度（<18天）。因此，分子改变很可能—— 通过持久的表观遗传修饰——这是 CBD 长期抗复发作用的基础。之前的研究 已经在来自均质组织的样本中发现了 CBD 引发的分子改变 伏核（NAc）——一个与药物复发密切相关的大脑部位。然而现有证据表明 线索触发的反应——例如药物渴望和寻求——是由“线索反应（激活）”介导的 而不是 NAc（和其他位点）中的“非反应性”神经元。鉴于只有一小部分（3-10%）神经元 NAc（和其他位点）对药物线索有反应，在源自样品的分子改变中发现 均质化组织——包含提示反应性和非反应性神经元以及非神经胶质细胞—— 可能与 CBD 的抗复发作用在功能上无关。相比之下，持久的表观遗传修饰 CBD 调节药物提示反应性的神经元所特有，可能介导长期的抗复发 CBD 的行动。基于上述科学前提，这个 R21 项目将检验以下假设： “CBD 通过药物提示反应神经元的表观遗传修饰发挥其持久的抗复发作用”。这 项目将采取多学科方法，包括 (1) 完善的可卡因动物模型 复发，(2) 一种用于活动依赖性神经标记的新型转基因大鼠品系，(3) 细胞类型特异性 通过荧光激活细胞分选进行分子采样，以及 (4) 下一代测序 表观遗传和转录标记的综合分析。主要目标是确定长期 CBD 对基因表达的持久影响——既具有细胞类型特异性又依赖于活性（依赖于药物） 线索反应性）——在表观基因组和蛋白质编码转录组水平上，从而有助于阐明 分子机制在功能上与 CBD 的长期抗复发作用有关。另一个主要目标是 建立 CBD 功能性表观遗传分析作为潜在抗复发的概念证明 药物治疗——一种可用于研究其他药物治疗作用的新方法。这 通过此类研究获得的知识可以用于提高医疗效果 精确识别脱靶效应。