Drug targeting the dynamics of opioid systems in alcohol dependence

项目总结 酒精使用障碍(AUD)是一种严重的疾病，具有严重的医学和社会后果。 在过去的几十年里，医疗方面几乎没有取得什么进展。我们正在采取翻译方法，并已 建立了一种动物模型，在该模型中，处于不同戒断阶段的酒精依赖大鼠是 调查。神经元的靶点是阿片系统和对手假说。我们建议最初的 兴奋效应是通过脑啡肽/u-阿片受体(MOP)和后来的发育来实现的。 负性强化(渴求)与强啡肽/kappa阿片受体(KOP)的活性有关。事实上， MOP拮抗剂纳曲酮是FDA批准的一种药物，有减少复发的适应症。KOP对手是 在不同的神经精神状态下进行临床试验。我们选择CERC-501作为指标药物， 可逆，在临床检查中明显耐受性良好。在行为记录之后，大脑样本将 被解剖并形成一个“脑库”，以供进一步分析。人们关注的焦点是杏仁核的中央核。 (CEA)，以及呈现MOP和/或KOP的电路。选定的标本将进行超分辨 显微镜(定量单分子定位显微镜，qSMLM)。一项初步研究表明， 急性剂量的乙醇扰乱MOP和KOP的定位，这一作用可被纳曲酮阻断。Etoh如何影响 细胞膜上的受体迁移率、受体聚集(同源和异源二聚体)及其与 富含蛋白质和脂肪的膜结构域将用荧光相关光谱(FCS)进行研究。AS 有了qSMLM，就可以在单分子水平上实现分辨率。这两种技术都将被用于研究联合 受体与与信号级联相关的蛋白质的定位。我们假设Etoh对 含有MOP和KOP的信号域的动态自组织，在 控制MOP和KOP组织的机制在慢性乙醇暴露过程中发展起来。使用OP拮抗剂 天然的MOP和KOP信号复合体在纳米水平上是稳定的。对这些的重点研究 机制将为乙醇诱导受体的分子机制提供关键的新见解 中断是可以防止或扭转的。这两种阿片系统的重要性可能不同。 并影响使用OP拮抗剂进行个性化治疗的选择。研究将包括 已知的N40D MOP基因会影响行为表型和对纳曲酮的敏感性。

PROJECT SUMMARY Alcohol use disorder (AUD) is a serious condition with severe medical and societal consequences.There has been little progress in medical treatment over the past decades. We are taking a translational approach and have established an animal model, where alcohol-dependent rats in various stages of abstinence are subjects of investigation. The neuronal target is the opioid systems and the opponent hypothesis. We propose that the initial euphoric effects are channeled through the enkephalin/mu-opioid receptor (MOP) and the later developing negative reinforcement (craving) is related to activity at the dynorphin/kappa-opioid receptor (KOP). In fact, the MOP antagonist, naltrexone is a FDA-approved agent with indication to reduce relapse. KOP antagonists are entering clinical trials in different neuropsychiatric conditions. We have chosen CERC-501 as index drug, being reversible and apparently well tolerated in clinical examination. After behavioral recordings, brain specimens will be dissected and form a “brain bank” for further analysis. A focus of interest is the central nucleus of the amygdala (CeA), and the circuitry presenting MOP and/or KOP. Selected specimens will undergo superresolution microscopy (quantitative Single Molecule Localization Microscopy, qSMLM). A pilot study showed that already an acute dose of EtOH disrupts localization of MOP and KOP, an effect blocked by naltrexone. How EtOH affects receptor mobility in the plasma membrane, receptor clustering (homo- and hetero-dimers) and association with protein- and lipid-rich membrane domains will be studied by fluorescence correlation spectroscopy (FCS). As with qSMLM resolution is achieved at the single-molecule level. Both technologies will be used to study co- localization of receptors with proteins of relevance for the signaling cascade. We hypothesize that EtOH perturbs the dynamic selforganization of signaling domains harboring MOPs and KOPs, that distinct alterations in mechanisms controlling MOP vs KOP organization develop during chronic EtOH exposure. With OP antagonists innate MOP and KOP signaling complexes are stabilized at the nanoscale level. Focused studies of these mechanisms will provide critical new insight into molecular mechanisms through which EtOH-induced receptor disruptions may be prevented or reversed. The commensurate importance of the two opioid systems may vary between individuals and influence the choice of personalized therapy with OP antagonists. Studies will include the N40D MOP genotype known to affect the behavioral phenotype and sensitivity to naltrexone.