THE ROLE OF RYANODINE RECEPTORS IN DRUG SEEKING

兰尼碱受体在寻求毒品中的作用

• 批准号: 10171831
• 负责人: ARTHUR C RIEGEL
• 金额: $ 38.36万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-15 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10171831
• 关键词: Acute; Adult; Antioxidants; Automobile Driving; Behavior; Behavioral; Biochemical; Brain; Calcium; Cells; Chronic; Cocaine; Cocaine Dependence; Cognitive; Cognitive deficits; Complex; Cues; Cyclic AMP-Dependent Protein Kinases; Data; Development; Dissociation; Dopamine; Drug Addiction; Electrophysiology (science); Extinction (Psychology); Extravasation; FKBP1B gene; Frequencies; Genetic Recombination; Glutathione; Goals; Heroin; Heroin Dependence; Hippocampus (Brain); Hypersensitivity; Image; Impaired cognition; Ion Channel; Knock-in Mouse; Laboratories; Link; Measures; Mediating; Modification; Molecular; Neurons; Nucleus Accumbens; Oxidation-Reduction; Oxidative Stress; Oxides; Pathologic; Pathology; Pathway interactions; Pharmaceutical Preparations; Pharmacological Treatment; Phosphorylation; Physiological; Play; Population; Post-Translational Protein Processing; Prefrontal Cortex; Proteins; Rattus; Reactive Oxygen Species; Regulation; Relapse; Role; Ryanodine Receptor Calcium Release Channel; Ryanodine Receptors; Self Administration; Signal Transduction; Slice; Specificity; Stress; Sucrose; Testing; Therapeutic; Wild Type Mouse; addiction; catalyst; cell type; cocaine relapse prevention; driving behavior; drug development; drug seeking behavior; effective therapy; glutathione S-transferase pi; hippocampal pyramidal neuron; in vivo; new therapeutic target; novel; oxidation; patch clamp; pre-clinical; prevent; reinforced behavior; small molecule; translational approach; treatment strategy

There currently exist no effective pharmacological treatment options to prevent relapse to cocaine and heroin seeking. This is due to a principal lack of understanding of the underlying maladaptive cellular mechanisms driving this behavior. Considerable evidence suggests that the pre-frontal cortex (PFC) projection to the nucleus accumbens core (NAcore) represents a principle common pathway for triggering relapse. Recently, we found in rats that cocaine or heroin SA induces persistent hyper-excitability in PFC pyramidal neurons. The hyperexcitability is related to elevated intracellular Ca2+ that likely arises from malfunctioning ryanodine receptors (RYRs). Excess Ca2+ suppresses inhibitory Kv7 ion channels that serve to limit neuronal firing. Notably, this drug-induced malfunction in cell signaling persists even after extended extinction of the drug- reinforced behavior. As a result, a subpopulation of PFC neurons remains hyper-excitable and likely hypersensitive to drug-associated cues. In hippocampal neurons, excessive stimulation by stress is associated with phosphorylation and oxidation of RYR2 and depletion of the stabilizing subunit calstabin2 (FKBP12.6) from the channel complex, resulting in intracellular Ca2+ leak through RYRs and cognitive dysfunction. A novel RYR-targeted small molecule Rycal (S107) that stabilizes the RYR2 closed states of PKA hyperphosphorylated, and oxidized/nitrosylated channels, prevents intracellular Ca2+leak and prevented the stress-induced cognitive defects. Our preliminary data indicate that S107 treatment also reduces cue-induced cocaine seeking in rats. The long-term goal of this project is to understand the cellular signaling and physiological mechanisms by which RYRs regulate relapse behavior in hopes of identifying better therapeutic strategies for the treatment of drug addiction. Our central hypotheses are that: (1) cocaine- and heroin- produces pathological destabilization of RYR via redox post-translational modification, 2) causing RYR- dependent Ca2+ leak within activated PFC neurons projecting to the nucleus accumbens core (NAcore), and 3) that the anti-relapse effects of the RYR stabilizer S107 are due, at least in part, to its ability to reduce RYR Ca2+ “leak”. To better understand the cellular origin of the enduring adaptations in PFC inhibition, we propose to examine: the redox state of RYR and a battery of biochemical changes in both RYR and FKBP12.6 in Aim 1, RYR changes in specific subpopulations of neurons in Aim 2, and lastly translational strategies aimed at reducing cued reinstatement of drug seeking in Aim 3. Together, these studies will extend our extensive preliminary findings that link the RYR redox state and the candidate addiction therapeutic, S107 to regulation of relapse-like behaviors that drive the long-lasting changes observed in the PFC. We will determine if disruption of the PFC RYR2 and/or FKBP12.6 by cocaine and heroin is necessary for relapse and will provide mechanistic preclinical data in support of a novel target for drug development to treat heroin and cocaine addiction.

目前尚无有效的药物治疗选择，无法防止可卡因和海洛因继电器 寻求。这是由于主要缺乏对潜在不良适应性细胞机制的理解 推动这种行为。大量证据表明，前额叶皮层（PFC）投影 伏隔核（Nacore）代表了触发继电器的主要共同途径。最近，我们 在可卡因或海洛因SA的大鼠中发现，在PFC锥体神经元中诱导了持续的高脱位性。这 过度兴奋与细胞内Ca2+的升高有关，这可能是由于ryanodine出现的 受体（RYRS）。Excess Ca2+抑制可限制神经元放电的抑制性KV7离子通道。 值得注意的是，这种药物诱导的细胞信号功能传导的故障，即使在药物扩展后仍存在 加强行为。结果，PFC神经元的亚群仍然是高度驱动的，并且可能 对药物相关线索的高度敏感。在海马神经元中，压力过度刺激与 随着RYR2的磷酸化和氧化和稳定亚基Calstabin2的耗竭（FKBP12.6） 从通道复合物，导致细胞内Ca2+通过RYRS和认知功能障碍。小说 稳定PKA的RYR2封闭状态 高磷酸化，氧化/硝基化通道可防止细胞内Ca2+泄漏，并防止 压力引起的认知缺陷。我们的初步数据表明S107处理也减少了提示诱导的 可卡因在老鼠中寻求。该项目的长期目标是了解细胞信号传导和 RYRS调节继电器行为以识别更好疗法的生理机制 治疗吸毒的策略。我们的中心假设是：（1）可卡因和海洛因 - 通过氧化还原后翻译后的修饰产生RYR的病理不稳定，2）导致Ryr- 投射到伏隔核（Nacore）的活化的PFC神经元内的依赖性CA2+泄漏，3） RYR稳定器S107的抗释放作用至少部分归因于其降低RYR的能力 CA2+“泄漏”。为了更好地了解PFC抑制中持久适应的细胞起源，我们提出了 检查：RYR的氧化还原状态以及AIM 1中RYR和FKBP12.6的一系列生化变化， RYR在AIM 2中神经元特定亚群的变化，最后翻译的策略旨在 减少在AIM 3中恢复毒品的提示恢复。这些研究将扩大我们的广泛 将RYR氧化还原状态与候选成瘾疗法联系起来的初步发现，S107与调节 在PFC中观察到的持久变化的浮雕样行为。我们将确定是否 可卡因和海洛因对PFC RYR2和/或FKBP12.6的破坏是救济的必要条件，将提供 机械临床前数据支持用于治疗海洛因和可卡因的新型药物开发目标 瘾。