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方案来预防可卡因和海洛因复发 寻求。这是由于缺乏对潜在的适应不良细胞机制的了解 推动这种行为。大量证据表明，前额皮质（PFC）向大脑的投射 伏隔核核心（NAcore）代表了触发复发的主要共同途径。最近，我们 在大鼠中发现可卡因或海洛因 SA 会诱导 PFC 锥体神经元持续过度兴奋。这 过度兴奋与细胞内 Ca2+ 升高有关，这可能是由兰尼碱故障引起的 受体（RYR）。过量的 Ca2+ 会抑制抑制性 Kv7 离子通道，从而限制神经元放电。 值得注意的是，这种药物引起的细胞信号传导故障即使在药物长期消失后仍然存在。 强化行为。因此，PFC 神经元的一个亚群仍然处于过度兴奋状态，并且可能 对药物相关线索过敏。在海马神经元中，压力的过度刺激与 RYR2 的磷酸化和氧化以及稳定亚基 calstabin2 (FKBP12.6) 的耗尽 通道复合体中的钙离子通过 RYR 泄漏，导致细胞内 Ca2+ 泄漏和认知功能障碍。一本小说 RYR 靶向小分子 Rycal (S107)，可稳定 PKA 的 RYR2 关闭状态 过度磷酸化和氧化/亚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 治疗还可以减少提示诱导的 在老鼠身上寻找可卡因。该项目的长期目标是了解细胞信号传导和 RYRs 调节复发行为的生理机制，希望找到更好的治疗方法 治疗毒瘾的策略。我们的中心假设是：（1）可卡因和海洛因 通过氧化还原翻译后修饰产生 RYR 的病理性不稳定，2) 导致 RYR- 激活的 PFC 神经元内依赖的 Ca2+ 渗漏投射到伏隔核核心 (NAcore)，以及 3) RYR 稳定剂 S107 的抗复发作用至少部分归因于其降低 RYR 的能力 Ca2+“泄漏”。为了更好地理解 PFC 抑制中持久适应的细胞起源，我们建议 检查：目标 1 中 RYR 的氧化还原状态以及 RYR 和 FKBP12.6 中的一系列生化变化， 目标 2 中特定神经元亚群的 RYR 变化，以及最后的翻译策略旨在 减少目标 3 中提示性的药物寻求恢复。总的来说，这些研究将扩展我们广泛的研究范围。 初步发现将 RYR 氧化还原状态和候选成瘾治疗药物 S107 与调节联系起来 导致 PFC 中观察到的长期变化的类似复发的行为。我们将确定是否 可卡因和海洛因对 PFC RYR2 和/或 FKBP12.6 的破坏是复发所必需的，并且会提供 支持治疗海洛因和可卡因药物开发新靶点的机械临床前数据 瘾。