Implementation of novel methodology to study the anti-relapse potential of cannabidiol

 描述（由申请人提供）：该探索性/开发性项目旨在建立新的方法学作为PI实验室的研究工具，以探索有趣的初步发现的神经生物学基础，即植物大麻素大麻二酚（CBD）减弱可卡因寻求，其效果显著超过治疗。导致可卡因成瘾的强迫性和慢性复发性的一个主要因素是由环境刺激引起的药物欲望，这些刺激已经成为可卡因主观效应的条件。在动物中，这些刺激物引起可卡因寻求的功效在很长一段时间内持续存在。 尽管在非强化条件下频繁接触，但仍有戒断期，反映了可卡因成瘾的强迫性。在初步研究中，CBD（大麻植物的主要非精神活性和非成瘾成分）显著减弱了持续性、强迫性样可卡因寻求动物模型的恢复，其作用在治疗终止后六周仍然没有减弱。这些研究结果表明，CBD逆转可卡因渴望和复发的神经可塑性，而不仅仅是短暂的药理学改善复发的脆弱性。因此，深入了解这些影响的机制可能对治疗药物的开发和理解强迫性可卡因寻求的神经和分子基础具有重要意义。该提案的目的是通过利用荧光激活细胞分选（FACS）和相关方法的最新进展来探索CBD持久的“抗恢复”作用的神经生物学基础。这些进展，由Co-I，Hope博士带头，允许快速高通量区域特异性识别（“神经映射”）编码特定行为的Fos表达神经元，同时以快速和定量的方式提供RNA，以允许表征来自单个大鼠的“行为激活”神经元的分子改变。该研究计划将扩展CBD对可卡因线索反应性的长期影响的探索，并在行为水平上寻求可卡因，并在实验室中建立FACS和相关方法，以确定与CBD寻求可卡因的持久衰减相关的大脑部位和基因表达。预计这些结果将为CBD发挥作用的神经和分子靶点提供重要的见解，并为随后的全面项目奠定基础，包括系统调查已确定的神经靶点的因果作用和基因表达变化介导CBD干扰可卡因寻求，利用流式细胞仪鉴定关键基因及其表观遗传调控机制，负责CBD作用的持久性，从而揭示预防复发的新的治疗靶点。

 DESCRIPTION (provided by applicant): This exploratory/developmental project is designed is to establish novel methodology as a research tool in the PI's laboratory to explore the neurobiological basis of intriguing preliminary findings that the phytocannabinoid cannabidiol (CBD) attenuates cocaine seeking with effects that significantly outlast treatment. A major factor contributing to the compulsive and chronically relapsing nature of cocaine addiction is drug desire elicited by environmental stimuli that have become conditioned to cocaine's subjective effects. In animals, the efficacy of these stimuli to elicit cocaine seeking perseverates over long periods of abstinence despite frequent exposure under non-reinforced conditions, reflective of the compulsive nature of cocaine addiction. In preliminary studies, CBD (the main non-psychoactive and non-addictive component of the cannabis sativa plant) significantly attenuated reinstatement in an animal model of perseverating, compulsive- like cocaine seeking, with effects that were still unabated six weeks after treatment termination. These findings suggest that CBD reverses neuroplasticity underlying cocaine craving and relapse beyond mere transient pharmacological amelioration of vulnerability to relapse. Insight into the mechanisms underlying these effects may therefore have major implications for treatment drug development and understanding of the neural and molecular basis of compulsive cocaine seeking. The objective of this proposal is to explore the neurobiological basis of CBD's lasting "anti-reinstatement" actions by exploiting recent advances in fluorescence-activated cell sorting (FACS) and associated methodologies. These advances, spearheaded by the Co-I, Dr. Hope, permit rapid high-throughput regionally specific identification ("neural mapping") of Fos-expressing neurons that encode specific behaviors while at the same time providing RNA in a rapid and quantitative manner to permit characterization of molecular alterations in "behaviorally activated" neurons from single rats. The research plan is to extend the exploration of CBD's long-lasting effects on responsiveness to cocaine cues and ensuing cocaine seeking at the behavioral level, and to establish FACS and associated methodologies in the lab to identify brain sites and gene expression linked to the lasting attenuation of cocaine seeking by CBD. The results are expected to provide essential insight into neural and molecular targets through which CBD exerts its actions and to lay the foundations for subsequent full-scale projects ranging from systematic investigation of causal roles of identified neural targets and gene expression changes in mediating CBD's interference with cocaine seeking, to utilization of FACS for the identification of key genes and their epigenetic regulatory mechanisms responsible for the persistence of CBD's actions, and thereby to reveal novel therapeutic targets for relapse prevention.