Circuit and epigenetic mechanisms underlying incubation of methamphetamine craving

甲基苯丙胺渴望潜藏的回路和表观遗传机制

• 批准号: 9751825
• 负责人: Xuan Anna Li
• 金额: $ 24.9万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751825
• 关键词: Abstinence; Amygdaloid structure; Area; Binding; Bioinformatics; Brain; ChIP-seq; Cholera Toxin Protomer B; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Cocaine; Corpus striatum structure; Cues; Data; Data Analyses; Development; Dopamine; Dorsal; Drug Addiction; Drug usage; Environment; Enzymes; Epigenetic Process; Exposure to; Family; Future; Gene Targeting; Genomics; Grant; HDAC5 gene; Heroin; Human; Incubated; Institution; Knock-out; Knowledge; Mediating; Mentors; Mentorship; Methamphetamine; Methamphetamine dependence; Methods; Modeling; Neurons; Pharmaceutical Preparations; Phase; Play; Prefrontal Cortex; Protocols documentation; RNA; Rattus; Recording of previous events; Relapse; Research Personnel; Research Proposals; Role; Self Administration; Substantia nigra structure; System; Techniques; Technology; Testing; Thalamic structure; Tracer; Training; Training Support; Transcript; Transcriptional Regulation; Viral; Withdrawal; activity marker; base; brain tissue; career; craving; designer receptors exclusively activated by designer drugs; disorder later incidence prevention; drug craving; drug relapse; drug withdrawal; insight; methamphetamine user; neuromechanism; novel; novel strategies; nuclease; overexpression; professor; receptor; tenure track; therapeutic target; transcriptome sequencing

Cue-induced drug craving progressively increases after withdrawal, a phenomenon termed ‘incubation of drug craving’. During the last decade, mechanistic studies have primarily focused on incubation of cocaine craving. In contrast, very few studies have examined the mechanisms underlying incubation of methamphetamine (Meth) craving. Here, I propose to study circuit and epigenetic mechanisms underlying incubation of Meth craving with a focus on the dorsal striatum (DS) region. The choice of this brain area is based on my preliminary data implicating a critical role of DS neuronal activity and a potential role of DS histone deacetylase 5 (HDAC5, an epigenetic-related enzyme) in incubation of Meth craving. At the circuit level, I will use a retrograde tracing technique (CTb, cholera toxin subunit B) in combination with the neuronal activity marker Fos to identify critical afferent projections to the DS that are activated during incubation of Meth craving. Next, I will use a novel retro-DREADD (Designer Receptors Exclusively Activated by Designer Drugs) method to study the causal role of afferent projections to the DS in incubation of Meth craving. At the epigenetic level, I will combine chromatin immunoprecipitation-sequencing (ChIP-seq) with RNA-sequencing (RNA-seq) to identify downstream gene targets of HDAC5 in DS whose expression is altered after prolonged withdrawal (1 month) from Meth self-administration. I will also use the CRISPR-Cas9 system to examine the causal role of DS HDAC5 in incubation of Meth craving. My proposal will provide new insights on the mechanisms underlying Meth craving and relapse. Additionally, under the mentorship of Drs. Shaham, Nestler, and Cowan the proposal will provide me with an ideal training environment to prepare me for a future independent career as a researcher in an academic setting who explores basic mechanisms of drug addiction.

戒断后，提示诱发的药物渴望逐渐增加，这种现象被称为“潜伏期” 毒瘾”。在过去的十年中，机制研究主要集中在可卡因的孵化上 渴望。相比之下，很少有研究探讨孵化的机制。 对甲基苯丙胺（Meth）的渴望。在这里，我建议研究潜在的电路和表观遗传机制 冰毒渴望的孵化，重点关注背侧纹状体（DS）区域。这个大脑区域的选择是基于 我的初步数据表明 DS 神经元活动的关键作用和 DS 组蛋白的潜在作用 脱乙酰酶 5（HDAC5，一种表观遗传相关酶）在冰毒渴望的孵化过程中。在电路层面，我将 使用逆行追踪技术（CTb，霍乱毒素 B 亚基）结合神经元活动 标记 Fos 来识别在冰毒渴望孵化期间激活的 DS 的关键传入投射。 接下来，我将使用一种新颖的retro-DREADD（Designer Receptors Exclusively Activated by Designer Drugs）方法来 研究对 DS 的传入投射在冰毒渴望孵化中的因果作用。在表观遗传层面，我会 将染色质免疫沉淀测序 (ChIP-seq) 与 RNA 测序 (RNA-seq) 相结合来鉴定 DS 中 HDAC5 的下游基因靶标，其表达在长期停药后（1 个月）发生改变 来自冰毒自我管理。我还将使用 CRISPR-Cas9 系统来检查 DS HDAC5 的因果作用 正在酝酿对冰毒的渴望。我的建议将为冰毒渴望的潜在机制提供新的见解 并复发。此外，在博士的指导下。 Shaham、Nestler 和 Cowan 该提案将提供 我拥有理想的培训环境，为我未来作为一名研究人员的独立职业做好准备 探索药物成瘾基本机制的学术环境。