Conditional Dicer1 manipulation to study miRNA involvement in opioid addiction

条件性 Dicer1 操作研究 miRNA 与阿片类药物成瘾的关系

• 批准号: 8322268
• 负责人: Gregory I Elmer
• 金额: $ 20.93万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8322268
• 关键词: Analgesics; Animal Model; Applications Grants; Architecture; Behavior; Behavioral; Bioinformatics; Biological; Brain region; Candidate Disease Gene; Chronic; Complex; Computer Simulation; Computing Methodologies; Databases; Dependence; Development; Disease; Dopamine; Drug Addiction; Drug Delivery Systems; Drug Exposure; Drug Tolerance; Emotional; Engineering; Enzymes; Family; Functional RNA; Gene Expression Profile; Genes; Genetic; Genetic Engineering; Genetic Techniques; Genetically Engineered Mouse; Genotype; Habits; Individual; Intake; Intervention; Intravenous; Investigation; Knowledge; Laboratories; Lead; Link; Mediating; Messenger RNA; Methods; MicroRNAs; Midbrain structure; Modeling; Molecular; Molecular Genetics; Molecular Profiling; Molecular Target; Morphine; Mus; Neuronal Differentiation; Neuronal Plasticity; Neurons; Opiate Addiction; Opioid; Opioid Analgesics; Pain; Pain management; Pathway interactions; Pattern; Pharmaceutical Preparations; Phenotype; Play; Process; Property; Psychological reinforcement; Rewards; Role; Self Administration; Societies; Staging; Surface; System; Techniques; Therapeutic; Therapeutic Intervention; Time; Ventral Striatum; addiction; adverse outcome; alternative treatment; approach behavior; brain tissue; cell type; clinically relevant; design; drug reward; economic cost; effective therapy; gamma-Aminobutyric Acid; high risk; insight; mRNA Expression; model design; neurobiological mechanism; neuropsychiatry; novel; novel strategies; novel therapeutics; promoter; reinforced behavior; research study; response; small hairpin RNA; trait; treatment strategy; Analgesics; Animal Model; Applications Grants; Architecture; Behavior; Behavioral; Bioinformatics; Biological; Brain region; Candidate Disease Gene; Chronic; Complex; Computer Simulation; Computing Methodologies; Databases; Dependence; Development; Disease; Dopamine; Drug Addiction; Drug Delivery Systems; Drug Exposure; Drug Tolerance; Emotional; Engineering; Enzymes; Family; Functional RNA; Gene Expression Profile; Genes; Genetic; Genetic Engineering; Genetic Techniques; Genetically Engineered Mouse; Genotype; Habits; Individual; Intake; Intervention; Intravenous; Investigation; Knowledge; Laboratories; Lead; Link; Mediating; Messenger RNA; Methods; MicroRNAs; Midbrain structure; Modeling; Molecular; Molecular Genetics; Molecular Profiling; Molecular Target; Morphine; Mus; Neuronal Differentiation; Neuronal Plasticity; Neurons; Opiate Addiction; Opioid; Opioid Analgesics; Pain; Pain management; Pathway interactions; Pattern; Pharmaceutical Preparations; Phenotype; Play; Process; Property; Psychological reinforcement; Rewards; Role; Self Administration; Societies; Staging; Surface; System; Techniques; Therapeutic; Therapeutic Intervention; Time; Ventral Striatum; addiction; adverse outcome; alternative treatment; approach behavior; brain tissue; cell type; clinically relevant; design; drug reward; economic cost; effective therapy; gamma-Aminobutyric Acid; high risk; insight; mRNA Expression; model design; neurobiological mechanism; neuropsychiatry; novel; novel strategies; novel therapeutics; promoter; reinforced behavior; research study; response; small hairpin RNA; trait; treatment strategy

DESCRIPTION (provided by applicant): Opioids are invaluable in pain management. Unfortunately, chronic opioid administration can lead to numerous adverse consequences such as a progressive decline in analgesic efficacy (tolerance), insurmountable pain and addiction. Thus, the extremely high therapeutic value of opioids is diminished by the detrimental impact of chronic opioid administration and abuse liability and can inflict enormous emotional and economic cost to individuals, families and society. Unfortunately, knowledge concerning the neurobiological mechanism associated with adverse consequences of chronic opioid administration has not produced an intervention or alternative treatment medication. A new approach is required to discover alternative treatment strategies. Our recent collaborative investigations have revealed a major role for canonical pathways involved in neuroplasticity and the discovery of microRNA (miRNA) involvement in morphine analgesic tolerance and drug self-administration. MiRNA expression is a strong candidate for coordinating the complex response to chronic drug exposure due to their network-like post-transcriptional effects on neuronal differentiation and dendritic architecture. Discovering the role of specific miRNA involvement in response to opioid self-administration may open the door to novel therapeutic strategies. The purpose of this CEBRA application is to define the role of miRNAs in self-administration using two novel strategies: 1) Genetically engineered mice with cell type-specific conditional knockdown of the rate- limiting miRNA processing enzyme, Dicer1, and 2) miRNA and mRNA expression profiling to discover miRNA:mRNA regulatory pairings involved in neuroadaptive changes. Our hypothesis is that escalated drug intake and the development of the self-administration habit is caused by a coordinated change in expression of select miRNAs. The following aims are proposed: Specific Aim 1: The purpose of this aim is to generate Cre-loxP animal models to enable timed and specific Dicer1 knockdown in dopaminergic and GA- BAergic neurons (using the Slc6a3 and Gad2 promotors, respectively) and phenotype self-administration behavior (and subsequent brain tissue) in a manner that can discriminate between active reinforcement and passive drug exposure. The results of this aim will provide valuable insight into miRNA's essential role, the cell- types involved and provide a phenotypic anchor for use in Aim 2. Specific Aim 2: Regional miRNA and mRNA expression profiles will be determined in multiple brain regions following morphine self-administration. The systematic process of expression profiling genotype-dependent behavioral responses to morphine and trait-specific associations/correlations with miRNA and mRNA expression will identify candidates specifically connected to self-administration. Computational methods will help prioritize miRNA:mRNA regulatory pairings and provide a functional framework to place the high-value pairs into targetable biological pathways. The multi- tiered, multi-disciplinary approach (behavior, molecular, bioinformatics) is designed to probe an unchartered layer of the genetic architecture (miRNA) to discover novel molecular targets for therapeutic intervention. PUBLIC HEALTH RELEVANCE: Opioid addiction inflicts enormous emotional and economic cost to individuals, families and society. Unfortunately, knowledge concerning the neurobiological mechanism associated with adverse consequences of chronic opioid administration has not produced an intervention or alternative treatment medication. A new focus is required to discover alternative treatment strategies--miRNAs provide a unique opportunity. The purpose of this grant application is to integrate of sophisticated behavioral techniques (yoked-control i.v. self-administration (SA) in the mouse), advanced genetic engineering (inducible cre-loxP system to delete Dicer1) and cutting edge molecular genetic techniques (miRNA expression profiling) to discover miRNA candidates involved in addiction and provide a discovery platform for truly novel treatment interventions.

描述（申请人提供）：阿片类药物在疼痛管理中是无价的。不幸的是，慢性阿片类药物给药会导致许多不利的后果，例如镇痛功效（耐受性），无法克服的疼痛和成瘾的逐渐下降。因此，阿片类药物的极高治疗价值因慢性阿片类药物给药和虐待责任的有害影响而降低，并可能给个人，家庭和社会带来巨大的情感和经济成本。不幸的是，关于与慢性阿片类药物的不利后果相关的神经生物学机制的知识尚未产生干预或替代治疗药物。需要一种新的方法来发现替代治疗策略。我们最近的合作研究揭示了涉及神经可塑性的规范途径的主要作用，并且发现了microRNA（miRNA）参与吗啡镇痛耐受性和药物自我给药。 miRNA表达是由于其网络样后对神经元分化和树突状结构的网络样作用，可以协调对慢性药物暴露的复杂反应的强大候选者。发现特定的miRNA参与对阿片类药物自我管理的作用可能打开了新型治疗策略的大门。 The purpose of this CEBRA application is to define the role of miRNAs in self-administration using two novel strategies: 1) Genetically engineered mice with cell type-specific conditional knockdown of the rate- limiting miRNA processing enzyme, Dicer1, and 2) miRNA and mRNA expression profiling to discover miRNA:mRNA regulatory pairings involved in neuroadaptive changes.我们的假设是，升级的药物摄入量和自我管理习惯的发展是由精选miRNA表达的协调变化引起的。提出了以下目的：具体目的1：此目的的目的是生成Cre-loxp动物模型，以在多巴胺能和gaergic神经元（分别使用SLC6A3和GAD2促进剂）和表型自动化行为（和随后的脑组织）中，在多巴胺能和gaer剂神经元中（分别使用SLC6A3和GAD2促进剂）中的定时和特定的DICER1敲低（分别使用SLC6A3和GAD2促进剂），以实现这种方式。该目标的结果将为miRNA的基本作用，涉及的细胞类型提供宝贵的见解，并提供了用于目标2的表型锚。特定目标2：区域miRNA和mRNA表达谱将在吗啡自加入后在多个大脑区域中确定。与miRNA和mRNA表达的形态依赖基因型依赖性行为反应的表达分析依赖性行为响应的系统过程将确定与自我管理专门连接的候选者。计算方法将有助于优先考虑miRNA：mRNA调节配对，并提供一个功能框架，以将高价值对放入目标生物学途径中。多层的多学科方法（行为，分子，生物信息学）旨在探测遗传结构（miRNA）未透明的层（miRNA），以发现用于治疗干预的新型分子靶标。 公共卫生相关性：阿片类药物成瘾对个人，家庭和社会造成了巨大的情感和经济成本。不幸的是，关于与慢性阿片类药物的不利后果相关的神经生物学机制的知识尚未产生干预或替代治疗药物。需要新的重点来发现替代治疗策略 - MIRNA提供了独特的机会。该赠款应用的目的是集成复杂的行为技术（鼠标中的i.v. i.v. i.v. i.v.自我给药（SA），高级基因工程（可诱导的CRE-loxp系统）以删除DICER1），并为了发现MirNA表达型候选者，并提供了一个涉及的概述，并提供了一个涉及MiRNA候选者的速度分子遗传技术（MiRNA表达分析）。