COCA: Project 1. Drug-induced ROS and Epigenetic Mechanisms

项目摘要-项目1考恩 治疗药物成瘾的一个主要挑战是对药物成瘾的分子机制缺乏了解。 吸毒会导致大脑功能的持续变化，即使在长期吸毒后，这种变化也会促进寻求毒品 禁欲。慢性药物使用涉及一个表观遗传过程，涉及染色质重塑酶， 组蛋白脱乙酰基酶5(HDAC5)，在纹状体中的刺神经元，其功能是限制发育。 多种与成瘾有关的行为，包括恢复吸食可卡因和寻求海洛因。然而， HDAC5的S在吸毒过程中和吸毒后的调节和作用是复杂的，人们对其知之甚少。可卡因和可卡因 海洛因产生活性氧和氮(ROS/RNS)，改变细胞的氧化还原状态，以及 氧化还原介导的半胱氨酸硫醇修饰改变靶蛋白的结构/功能，包括IIa类 HDAC。动物或人类反复使用抗氧化剂和谷胱甘肽前体N- 乙酰半胱氨酸(NAC)，减少恢复可卡因和海洛因寻找的易感性，以及NAC 阻止ROS推动的HDAC5核出口。项目1的长期目标是了解氧化还原 HDAC5和NAC调控海洛因和可卡因复发的蛋白质信号和表观遗传学机制 希望找到治疗药物成瘾的更好的治疗策略。我们 假设：(1)可卡因和海洛因产生的氧化应激改变HDAC5半胱氨酸硫基 伏隔核，促进核输出，减少HDAC5的抗复发作用，以及 (2)反复NAC治疗的抗复发效果至少部分归因于其保护HDAC5的能力 来自促进核排斥和基因表达变化的ROS信号事件。我们将使用 尖端的小鼠遗传学和依赖于cre的病毒工具，允许时间分隔和细胞类型特定 在吸毒和复发事件中对伏核核心神经元的操纵。 目标1.分子和细胞机制：在这个目标中，我们将确定可卡因和海洛因SA如何 调控HDAC5‘S氧化还原敏感的翻译后修饰、亚细胞分布、基因组结合和 基因表达的调控。 目的2.电路特异性：在这个目标中，我们将采用前沿的分子遗传学方法来剖析 HDAC5负性调控可卡因和海洛因毒品寻找的细胞类型特异性作用。 目的3.治疗：在这个目标中，我们将分析NAC对HDAC5的调节，并测试HDAC5的作用 NAC在减少海洛因和可卡因寻觅的恢复方面的长期能力。

PROJECT SUMMARY – Project 1 Cowan A major challenge for treating drug addiction is the poor understanding of the molecular mechanisms by which drug use produces persistent changes in brain function that facilitate drug seeking even after long periods of abstinence. Chronic drug use engages an epigenetic process, involving the chromatin-remodeling enzyme, histone deacetylase 5 (HDAC5), in striatal medium spiny neurons that functions to limit the development of multiple addiction-related behaviors, including reinstatement of cocaine and heroin seeking. However, HDAC5’s regulation and function during and after drug taking is complex and poorly understood. Cocaine and heroin produce reactive oxygen and nitrogen species (ROS/RNS) that alter the redox state of the cell, and redox-mediated cysteine thiol modifications alter the structure/function of target proteins, including class IIa HDACs. Repeated treatment of animals or humans with the antioxidant and glutathione precursor, N- acetylcysteine (NAC), reduces the vulnerability to reinstatement of cocaine and heroin seeking, and NAC blocks ROS-promoted HDAC5 nuclear export. The long-term goal of Project 1 is to understand the redox protein signaling and epigenetic mechanisms by which HDAC5 and NAC regulate heroin and cocaine relapse behaviors in hopes of identifying better therapeutic strategies for the treatment of drug addiction. We hypothesize that: (1) cocaine- and heroin-produced oxidative stress modifies HDAC5 cysteine thiol groups in the nucleus accumbens, which promotes nuclear export and reduces the anti-relapse actions of HDAC5, and (2) the anti-relapse effects of repeated NAC treatment are due, at least in part, to its ability to protect HDAC5 from ROS signaling events that promote nuclear exclusion and changes in gene expression. We will use cutting-edge mouse genetics and cre-dependent viral tools that allow for time-delimited and cell-type specific manipulation of nucleus accumbens core neurons during drug taking and relapse events. Aim 1. Molecular and Cellular Mechanisms: In this aim, we will determine how cocaine and heroin SA regulate HDAC5’s redox-sensitive posttranslational modifications, subcellular distribution, genomic binding and regulation of gene expression. Aim 2. Circuit Specificity: In this aim, we will take a cutting-edge molecular genetic approach to dissect the cell-type specific roles for HDAC5 to negatively-regulate cocaine and heroin drug seeking. Aim 3. Treatment: In this aim, we will analyze the regulation of HDAC5 by NAC, and test the role of HDAC5 in the long-lasting ability of NAC to reduce reinstatement of heroin and cocaine seeking.