Functional Epigenetic Profiling of Anti-Relapse Cannabidiol

药物成瘾是一种慢性复发性的强迫性渴求、寻求和使用药物的障碍。我们已经确定 大麻二酚（CBD）的实质性治疗潜力-一种天然存在于 大麻植物-防止药物复发。通过临床相关的经皮给药对CBD进行7天治疗 路线减少了提示和压力触发的可卡因恢复以及大鼠的酒精寻求-以及- 建立药物复吸动物模型。结合阿片类药物的相关报告，CBD似乎 有效对付三大类药物的两种主要复吸促进模式。 虽然潜在的神经生物学机制尚不清楚，CBD的抗复发作用（在138 治疗后30天）大大超过其生物利用度（<18天）。因此很可能是分子改变- 通过持久的表观遗传修饰-CBD的长期抗复发作用的基础。以前的研究 已经确定了CBD触发的分子变化的样品来源于匀浆组织的 脑桥核（NAc）-与药物复发密切相关的脑部位。然而，现有证据表明， 线索触发的反应--如药物渴望和寻求--是由“线索反应（激活）”介导的， 而不是NAc（和其他部位）中的“非反应性”神经元。鉴于只有一小部分（3-10%）神经元 在NAc（和其他网站）是反应性的药物线索，分子改变中确定的样品来源于 均质化组织-包含线索反应性和非反应性神经元以及非神经胶质， 可能在功能上与CBD的抗复发作用无关。相反，持久的表观遗传修饰 独特的神经元，其中药物线索反应性是由CBD调制可能介导延长抗复发 CBD的行动。基于上述科学前提，R21项目将测试以下假设： CBD通过药物线索反应神经元的表观遗传修饰发挥其长期的抗复发作用。的 该项目将采取多学科方法，包括：（1）一个成熟的可卡因动物模型 复发，（2）活动依赖性神经标记的转基因大鼠新品系，（3）细胞类型特异性 通过荧光激活细胞分选进行分子取样，和（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-细胞类型特异性和活性依赖性（对药物 线索反应性）-在表观基因组和蛋白质编码转录组的水平，从而帮助阐明 与CBD的长期抗复发作用功能相关的分子机制。另一个主要目标是 建立CBD作为潜在抗复发药物的功能性表观遗传分析的概念验证 药物-一种新的方法，可用于研究其他药物的治疗作用。的 通过这些研究获得的知识然后可以用于通过以下方式来提高医学治疗的功效： 精确识别靶上效应和脱靶效应。

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.