MicroRNA-mediated mechanisms of heroin drug seeking

MicroRNA介导的海洛因毒品寻找机制

• 批准号: 9981715
• 负责人: STEPHANIE Sillivan
• 金额: $ 23.89万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9981715
• 关键词: Abstinence; Address; Alcohol or Other Drugs use; Alcohols; Amygdaloid structure; Animal Model; Animals; Behavior; Behavioral; Brain; Brain region; Cocaine; Complex; DNA Methylation; Data; Data Set; Disease model; 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; Opioid agonist; Outcome; Overdose; Pathway interactions; Peptide Sequence Determination; Pharmaceutical Preparations; Pharmacology; Phase; 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; bioinformatics tool; comorbidity; cost; craving; drug craving; drug relapse; drug seeking behavior; druggable target; effective therapy; experience; experimental study; flexibility; frontal lobe; in vivo; insight; methylation pattern; miRNA expression profiling; mouse model; new therapeutic target; novel; opioid use; opioid withdrawal; prevent; programs; public health relevance; skills; substance use prevention; transcriptome sequencing; traumatic stress

 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（由申请方提供）：流行病学趋势表明，近年来，高度成瘾的阿片类海洛因滥用和过量的发生率增加了两倍多。然而，缺乏有效的治疗选择，这反映在阿片类药物复吸和过量的高比率上。这个问题的核心在于药物渴望在阿片类药物脱毒后长期持续甚至加强的能力。近年来，这种“渴望孵化”效应的行为机制已经得到了很好的表征，但需要更多的分子洞察力来确定大脑中的可药物靶点，这些靶点在长期禁欲后维持药物寻求。为此，该项目的目标是确定维持强烈药物寻求的机制，由microRNA（miRNA）介导的翻译机制有助于长期禁欲后继续寻求海洛因的假设指导。虽然它们在阿片寻求中的作用尚未描述，但miRNA是本研究的理想焦点，因为每个miRNA都可以靶向数百个基因，使单个miRNA具有高度的机械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，例如阿片激动剂的调节以及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的培训，以确定miRNA-蛋白质靶点相互作用。目前，候选人正在研究miRNAs在创伤后应激障碍（PTSD）小鼠模型中创伤记忆储存中的作用，并在初步数据中采用小RNA测序（miRNA-Seq）来测量创伤学习经历后持续的miRNA表达（>30天）。在指导K99期间，当前项目的范围将扩展到进行蛋白质组学分析，并将其与现有的miRNA-Seq数据集进行比对，从而能够识别miRNA通路，这些通路将在PTSD模型中进行功能测试，以支持创伤应激记忆。在R 00期间，候选人将应用所获得的miRNA和阿片类药物学的技能和知识，使用海洛因自我给药的啮齿动物模型研究海洛因渴望孵化的miRNA机制。集中在两个大脑区域参与阿片类药物渴求的孵化，中央杏仁核和眶额皮质，miRNA的表达及其通路将在体内进行操作，以功能性地识别在海洛因戒断30天后维持药物寻求的miRNA机制。该项目的完成将确定无毒品大脑中阿片类药物寻求的新机制，这是预防物质使用复发的关键一步。