Drug targeting the dynamics of opioid systems in alcohol dependence

项目摘要 酒精使用障碍（AUD）是一种严重的疾病，具有严重的医疗和社会后果。 在过去的几十年里，医疗方面几乎没有进展。我们正在采取一种转化的方法， 建立了一个动物模型，在不同阶段的戒酒酒精依赖大鼠的主题， 调查神经元靶点是阿片系统和对手假说。我们建议， 欣快效应是通过脑啡肽/μ阿片受体（MOP）和后来发展的 负强化（渴望）与强啡肽/κ-阿片受体（KOP）的活性有关。实际上 MOP拮抗剂纳洛酮是FDA批准的用于减少复发的药物。KOP拮抗剂是 在不同的神经精神疾病中进行临床试验。我们选择CERC-501作为索引药物， 在临床检查中可逆且明显耐受良好。在行为记录之后，大脑样本将 被解剖并形成一个“大脑银行”以供进一步分析。杏仁核的中央核是一个研究的焦点 (CeA)以及呈现MOP和/或KOP的电路。选定的标本将进行超分辨率 显微镜（定量单分子定位显微镜，qSMLM）。一项试点研究表明， 急性剂量的EtOH破坏MOP和KOP的定位，这种作用被纳洛酮阻断。EtOH如何影响 质膜中的受体迁移率、受体聚集（同源和异源二聚体）和与 将通过荧光相关光谱（FCS）研究富含蛋白质和脂质的膜区域。作为 在单分子水平上实现了qSMLM分辨率。这两项技术将用于研究合作， 用与信号级联相关的蛋白定位受体。我们假设乙醇干扰了 携带MOPs和KOP的信号结构域的动态自组织， 在慢性EtOH暴露期间，控制MOP与KOP组织的机制发展。使用OP拮抗剂 先天MOP和KOP信号传导复合物在纳米级水平上稳定。重点研究这些 机制将提供关键的新的洞察分子机制，通过EtOH诱导的受体 可以防止或逆转中断。两种阿片系统的同等重要性可能有所不同 个体之间的差异，并影响OP拮抗剂个性化治疗的选择。研究将包括 已知影响行为表型和对纳洛酮敏感性的N40 D MOP基因型。

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.