Cannabidiol: Lasting attenuation of ethanol seeking

 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，探索有趣的初步发现的神经生物学基础，即用植物大麻素大麻二醇短暂治疗显著减少乙醇(EtoH)寻求，其效果比治疗持续几个月。酒精中毒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酒精欲望，这些刺激已经成为药物主观效果的条件。事实上，这些刺激诱导乙醇寻求的有效性在长时间的禁欲中持续存在，尽管经常暴露在非强化条件下，这反映了酒精中毒的强迫性质。在初步研究中，大麻二酚(CBD)，大麻植物的主要非精神活性和非成瘾成分，显著减弱了强迫症样持续寻求乙醇的动物模型的恢复，其效果在治疗终止五个月后仍未减弱。这些发现表明，CBD逆转了与潜在的乙醇渴求和复发相关的适应不良学习的神经可塑性，而不仅仅是短暂的药物改善复发的易感性。因此，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乙醇寻求的神经和分子基础具有重要意义。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对乙醇线索的反应和随后的乙醇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持久减弱乙醇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重要的洞察力，并为后续的全面项目奠定基础，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介导CBD对Etoh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 brief treatment with the phytocannabinoid cannabidiol significantly reduces ethanol (EtOH) seeking with effects that outlast treatment by several months. A major factor contributing to the compulsive and chronically relapsing nature of alcoholism is EtOH desire elicited by environmental stimuli that have become conditioned to the drug's subjective effects. Indeed, the efficacy of these stimuli to elicit EtOH seeking perseverates over long periods of abstinence despite frequent exposure under non-reinforced conditions, reflective of the compulsive nature of alcoholism. In preliminary studies, cannabidiol (CBD), the main non-psychoactive and non-addictive component of the cannabis sativa plant significantly attenuated reinstatement in an animal model of compulsive-like perseverating EtOH seeking, with effects that were still unabated five months after treatment termination. These findings suggest that CBD reverses neuroplasticity linked to maladaptive learning underlying EtOH 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 EtOH 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 EtOH cues and ensuing EtOH 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 EtOH 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 EtOH 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.