Translational and epigenetic profiling of cell types associated with addiction

与成瘾相关的细胞类型的翻译和表观遗传分析

基本信息

批准号：
7938631
负责人：
NATHANIEL HEINTZ
金额：
$ 231.35万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7938631
关键词：
Address Affinity Chromatography Amygdaloid structure Animals Antibodies Area Behavior Biological Assay Brain region Cell Nucleus Cells Central Nervous System Diseases Cocaine Corpus striatum structure DNA Data Data Analyses Data Set Development Drug Addiction Epigenetic Process Euchromatin Event Follow-Up Studies Gene Expression Genes Genetic Genome Genomics Goals Grant Histones Interneurons Intervention Knowledge Laboratories Maps Measures Methamphetamine Methodology Methylphenidate Modification Molecular Molecular Neurobiology Molecular Profiling Mus National Institute of Drug Abuse Neurons Nucleus Accumbens Population Property Regulation Research Ribosomes Ritalin Site Spinal Structure Substance abuse problem Thalamic structure Transgenic Mice Translating United States abstracting addiction cell type cholinergic chromatin immunoprecipitation cocaine exposure comparative cost drug addiction therapy drug seeking behavior frontal lobe in vivo meetings neural circuit novel programs protein profiling public health relevance response success

项目摘要

DESCRIPTION (provided by applicant): Project Summary/Abstract: Recent studies of addiction have highlighted several regions of the brain that are thought to be involved in goal directed and drug seeking behaviors. The specific neuronal classes involved in the regulation of these behaviors are beginning to be identified, and attempts to profile the molecular changes in these cell types occurring as a consequence of addiction have met with some success. While these studies demonstrate that it is possible to discover changes in cell types that are correlated with, and in some cases contribute to, addiction, our knowledge in this area is fragmentary and incomplete. This is due to technical obstacles that have faced this field for decades, and that have recently been overcome by novel methodologies developed in our laboratories. The objective of this program of research is to identify all of the changes in gene expression and accompanying alterations in epigenetic regulation that contribute to the addictive state in cell types known to be important in the neural circuitry controlling addiction. The approach we will take in this program is to: 1) employ TRAP methodology and bacTRAP transgenic mouse lines to comprehensively profile the translated mRNAs from fifteen mouse CNS cell types that are components of the neural circuitry controlling addiction; 2) to collect translational profiles from each of these cell types in mice exposed to cocaine, methylphenidate, and methamphetamine 3) perform in depth comparative analysis of these resulting microarray datasets to identify changes in gene expression that accompany the addictive state in each cell type; 4) to concurrently isolate nuclei from each cell type and map cell specific sites of mC and hmC modification to the neuronal genomes; 5) to conduct follow up studies on genomic loci identified in the preceding aims to map additional epigenetic regulatory events by ChIP assays using H3K9m2 and H3K27m3 specific antibodies to identify genomic loci associated with suppression of gene expression in euchromatin. This project will provide information and experimental animals that will stimulate addiction research in a broad spectrum of laboratories, and provide materials and a paradigm for comprehensive studies of these same neural circuits under other experimental conditions. As such, this program will have an enormous impact on modern molecular neurobiology, and on the development of novel targets for pharmacological interventions in CNS disorders. PUBLIC HEALTH RELEVANCE: According to the National Institute on Drug Abuse, "Estimates of the total overall costs of substance abuse in the United States ..... exceed half a trillion dollars annually". The goals of the project are to comprehensively profile molecular changes occurring in specific cell types in regions associated with addiction, and to concurrently map epigenetic changes occurring in these cell types. It will stimulate in depth research into cell specific molecular events that are responsible for establishment of the addictive state, and identify novel targets for the development of new therapies for drug addiction.

描述（由申请人提供）：项目摘要/摘要：成瘾的最新研究强调了大脑的几个区域，这些区域被认为与目标定向和寻求药物行为有关。这些行为调节所涉及的特定神经元类已开始识别，并且试图介绍由于成瘾而发生的这些细胞类型中分子变化的尝试已经取得了一些成功。尽管这些研究表明，有可能发现与成瘾相关的细胞类型的变化，而在某些情况下，我们在这一领域的知识是零碎和不完整的。这是由于数十年来面临该领域的技术障碍，并且最近通过我们的实验室中开发的新方法克服了这种障碍。该研究计划的目的是确定基因表达的所有变化以及表观遗传调节的变化，这些变化在控制成瘾的神经回路中已知重要的细胞类型中有助于成瘾状态。我们将采用该程序的方法是：1）采用陷阱方法和Bactrap转基因小鼠系列来全面介绍来自15个小鼠CNS细胞类型的翻译mRNA，这些mRNA是控制成瘾的神经回路的组成部分； 2）在暴露于可卡因，甲基甲酯和甲基苯丙胺的小鼠中，从每种细胞类型中收集翻译曲线3）对这些产生的微阵列数据集进行了深入的比较分析，以识别每种细胞类型中成瘾状态的基因表达变化； 4）同时分离核与MC和HMC修饰的每种细胞类型的特异性位点分离到神经元基因组； 5）在先前的目的中对基因组基因座进行后续研究，旨在通过使用H3K9M2和H3K27M3特异性抗体来绘制其他表观遗传调节事件，以鉴定与抑制抑制基因表达在呼吸素中的基因组基因瘤相关的基因组基因瘤。该项目将提供信息和实验性动物，这些动物将在各种实验室中刺激成瘾研究，并为在其他实验条件下对这些相同的神经回路进行全面研究提供材料和范式。因此，该程序将对现代分子神经生物学以及中枢神经系统疾病的药理干预措施的新靶标有巨大影响。公共卫生相关性：根据美国国家药物滥用研究所的说法，“估计美国药物滥用的总成本.....每年超过半万亿美元”。该项目的目标是全面介绍与成瘾相关区域中特定细胞类型中发生的分子变化，并同时映射这些细胞类型中发生的表观遗传变化。它将刺激对细胞特定分子事件的深入研究，这些事件负责建立成瘾状态，并确定新的药物成瘾疗法的新目标。