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

 描述(申请人提供)：这个探索性/开发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醇(CBD)抑制可卡因的寻找，其效果显著超过治疗。造成可卡因成瘾的强迫性和长期复发性的一个主要因素是由环境刺激引起的毒品欲望，这些环境刺激已经习惯于可卡因的主观影响。在动物身上，这些刺激诱导寻求可卡因的效果会持续很长时间。 尽管经常暴露在非强化条件下，但仍有一段禁欲时期，这反映了可卡因成瘾的强迫性。在初步研究中，CBD(大麻植物中主要的非精神活性和非成瘾成分)显著减弱了坚持、强迫性类可卡因寻求的动物模型的恢复，其效果在治疗终止六周后仍未减弱。这些发现表明，CBD逆转了潜在的可卡因渴望和复发的神经可塑性，而不仅仅是短暂的药物改善复发的易感性。因此，深入了解这些效应的潜在机制可能对治疗、药物开发和了解强迫性寻求可卡因的神经和分子基础具有重大意义。这项建议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“反恢复”行动的神经生物学基础。由Co-I的Hope博士带头的这些进展，可以对编码特定行为的Fos表达的神经元进行快速的高通量区域特异性鉴定(“神经图谱”)，同时以快速和定量的方式提供RNA，以便能够表征单个大鼠的“行为激活”神经元的分子变化。这项研究计划在行为水平上扩展CBD对可卡因线索的反应性和随后的可卡因寻求的长期影响的探索，并在实验室建立流式细胞仪和相关方法学，以确定与CBD持久减弱可卡因寻求有关的大脑部位和基因表达。这些结果有望为CBD发挥作用的神经和分子靶点提供必要的洞察，并为后续的全面项目奠定基础，范围从系统研究已识别的神经靶点的因果作用和基因表达变化调节CBD对可卡因寻找的干扰，到利用流式细胞仪识别CBD持续作用的关键基因及其表观遗传调控机制，从而揭示预防复发的新治疗靶点。

 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.