MicroRNA-mediated mechanisms of heroin drug seeking

MicroRNA介导的海洛因毒品寻找机制

• 批准号: 9259957
• 负责人: STEPHANIE Sillivan
• 金额: $ 16.02万
• 依托单位: SCRIPPS FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9259957
• 关键词: Abstinence; Address; Agonist; Alcohol or Other Drugs use; Alcohols; Amygdaloid structure; Animal Model; Animals; Behavior; Behavioral; Bioinformatics; Brain; Brain region; Cocaine; Complex; DNA Methylation; Data; Data Set; Disease model; Drug Metabolic Detoxication; Drug usage; Epidemiological trend; Fright; Funding; Future; Genes; Genetic Transcription; Genomics; Goals; Gold; Grant; Heroin; Human; Incidence; Individual; Knowledge; Learning; Maintenance; Mass Spectrum Analysis; Measures; Mechanics; Mediating; Mediation; Memory; Mentors; Methodology; MicroRNAs; Modeling; Modification; Molecular; Motivation; Mus; Opioid; Outcome; Overdose; Pathway interactions; Peptide Sequence Determination; Pharmaceutical Preparations; Pharmacology; Phase; Plasticizers; Post-Traumatic Stress Disorders; Pre-Clinical Model; Process; Proteins; Proteome; Proteomics; Protocols documentation; Regulation; Rehabilitation therapy; Relapse; Research; Rodent Model; Role; Self Administration; Set protein; Small RNA; Solid; Stress; Testing; Tissues; Training; Withdrawal; Work; addiction; behavioral study; cost; craving; drug craving; drug relapse; drug seeking behavior; effective therapy; experience; experimental study; flexibility; frontal lobe; in vivo; insight; methylation pattern; mouse model; new therapeutic target; novel; opioid use; prevent; programs; public health relevance; skills; substance use prevention; tool; transcriptome sequencing

 DESCRIPTION (provided by applicant): Epidemiological trends indicate that the incidence of abuse and overdose of the highly addictive opioid heroin have more than tripled in recent years. However, effective treatment options are lacking and this is reflected in the high rates of opioid relapse and overdose. At the center of this issue lies the ability of drug cravings to persist and even strengthen long after opioid detoxification. The behavioral mechanics of this "incubation of craving" effect have become well-characterized in recent years, but more molecular insight is required to identify druggable targets within the brain that sustain drug seeking after prolonged abstinence. To that end, the goal of this project is to identify mechanisms that sustain strong drug seeking, guided by the hypothesis that microRNA (miRNA)-mediated translational mechanisms contribute to continued heroin seeking after prolonged abstinence. While their role in opioid seeking has not been described, miRNAs are an ideal focus for this study because each miRNA can target hundreds of genes, giving a single miRNA a high degree of mechanistic flexibility and a wide genomic range, capable of orchestrating complex behaviors. Indeed, limited exploration into their role in addiction has yielded exciting and promising results, such a regulation by opioid agonists and modification of alcohol and cocaine seeking. To address the goal of the project, the mentored phase of the grant will be used to gain training in heroin self-administration, protein sequencing and bioinformatic tools to identify miRNA-protein target interactions. At present, the candidate is studying the role of miRNAs in traumatic memory storage in a mouse model of post-traumatic stress disorder (PTSD) and in preliminary data employed small RNA sequencing (miRNA-Seq) to measure lasting miRNA expression (>30 days) after a traumatic learning experience. During the mentored K99 period, the scope of the current project will be extended to perform proteomic analysis and align it with the existing miRNA-Seq dataset, enabling the identification of miRNA pathways that will be functionally tested for their support of traumatic stress memories in the PTSD model. In the R00 period, the candidate will then apply the acquired skills and knowledge of miRNAs and opioid pharmacology to study miRNA mechanisms in the incubation of heroin craving using a rodent model of heroin self-administration. Focusing on two brain regions involved in the incubation of opioid craving, the central amygdala and orbitofrontal cortex, expression of miRNAs and their pathways will be manipulated in vivo to functionally identify miRNA mechanisms that sustain drug seeking after 30 days of abstinence from heroin. Completion of this project will identify new mechanisms of opioid seeking in the drug- free brain, which represents a critical step towards preventing substance use relapse.

 描述（由申请人提供）：流行病学趋势表明，高度成瘾的阿片类海洛因滥用和过量服用的发生率近年来增加了两倍多。然而，缺乏有效的治疗方案，这反映在阿片类药物复发和过量用药的高发生率上。这个问题的核心在于药物渴望在阿片类药物戒毒后很长时间内持续存在甚至加强。近年来，这种“渴望的孵化”效应的行为机制已得到充分表征，但需要更多的分子洞察力来识别大脑内可在长期禁欲后维持药物寻找的可药物靶标。为此，该项目的目标是确定维持强烈药物寻求的机制，该假设以 microRNA (miRNA) 介导的翻译机制有助于长期戒毒后继续寻求海洛因为指导。虽然它们在阿片类药物寻求中的作用尚未被描述，但 miRNA 是这项研究的理想焦点，因为每个 miRNA 都可以靶向数百个基因，使单个 miRNA 具有高度的机制灵活性和广泛的基因组范围，能够协调复杂的行为。事实上，对它们在成瘾中的作用的有限探索已经产生了令人兴奋和有希望的结果，例如阿片类激动剂的调节以及对酒精和可卡因寻求的改变。为了实现该项目的目标，赠款的指导阶段将用于获得海洛因自我管理、蛋白质测序和生物信息学工具方面的培训，以识别 miRNA-蛋白质靶标相互作用。目前，该候选人正在创伤后应激障碍（PTSD）小鼠模型中研究miRNA在创伤记忆存储中的作用，并在初步数据中采用小RNA测序（miRNA-Seq）来测量创伤性学习经历后持久的miRNA表达（>30天）。在 K99 指导期间，当前项目的范围将扩大到进行蛋白质组分析，并将其与现有的 miRNA-Seq 数据集进行对齐，从而能够识别 miRNA 通路，这些通路将进行功能测试，以支持 PTSD 模型中的创伤应激记忆。在 R00 阶段，候选人将应用所获得的 miRNA 和阿片类药物药理学的技能和知识，使用海洛因自我给药的啮齿动物模型来研究 miRNA 在海洛因渴望孵化中的机制。重点关注参与阿片类药物渴望孵化的两个大脑区域，即中央杏仁核和眶额皮质，将在体内操纵 miRNA 的表达及其途径，以功能性地识别在戒除海洛因 30 天后维持药物寻求的 miRNA 机制。该项目的完成将确定在无药物的大脑中寻找阿片类药物的新机制，这是防止药物滥用复发的关键一步。