Next Generation Sequencing of Human Alcoholic Brain

人类酒精大脑的下一代测序

基本信息

批准号：
8912956
负责人：
R. DAYNE MAYFIELD
金额：
$ 29.27万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-05 至 2017-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8912956
关键词：
Address Alcohol abuse Alcohol consumption Alcohol dependence Alcohols Alternative Splicing Amygdaloid structure Animal Model Anterior Autopsy Award Biological Biological Process Brain Brain region Cell physiology Chromosome Mapping Classification Clinical Code Collaborations Complex Cromolyn Sodium DNA Microarray Chip Data Dependence Detection Diagnosis Dorsal Event Gene Expression Gene Expression Profile Gene Structure Genes Genome Genomics Goals Heavy Drinking Human Human body Indiana Individual Kinetics Knowledge Life Macaca Medial Mediating Medical Methodology Methods MicroRNAs Microarray Analysis Modeling Molecular Molecular Profiling Mus National Institute on Alcohol Abuse and Alcoholism Neurosciences Outcome Pathway Analysis Pattern Pilot Projects Population Prefrontal Cortex Price Property Publishing RNA RNA Splicing Reporting Research Personnel Reverse Transcriptase Polymerase Chain Reaction Rewards Rodent Sampling Signal Transduction Single Nucleotide Polymorphism Site Small RNA Systems Biology Technology Testing Tissues Transcript Universities Untranslated RNA Variant Weight Work alcohol exposure alcohol research base cDNA Arrays cingulate cortex cost differential expression drug of abuse drug reward functional group human disease human subject human tissue innovation interest new technology next generation next generation sequencing novel problem drinker research study targeted sequencing tissue resource transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): We hypothesize that long-term alcohol abuse results in changes in gene expression in brain and that these changes are responsible, at least partly, for alcohol dependence. We also hypothesize that there is significant structural variation in expression patterns of specific genes that can be detected by next generation sequencing. The goal of this project is to utilize this state-of-the-art technology to identify novel transcripts that are differentially expressed in brain regions Involved in drug reward of well-characterized human subjects as a function of alcohol intake, abuse, and dependence. To date, there are no published reports of gene expression in the human extended amygdala and related cortical projection regions. The results of these studies will identify as-yet-unknown alcohol responsive genes, including low abundance, splice variants, and expressed SNPs. In collaboration with other INIA investigators, we will have the opportunity to analyze and compare sequence data obtained from human, macaque, and rodents following alcohol exposure. The proposed experiments are a direct outcome of a prior INIA West Pilot Project awarded to the PI. Three Specific Aims are proposed: Aim 1 will test the hypothesis that next generation transcriptome profiling (RNA-seq) will identify differentially expressed RNAs in select brain regions of well-characterized alcohol-dependent cases compared to matched controls. Subsequent RT-PCR analysis will validate and define which of these are involved in mediating the effects of alcohol consumption. Aim 2 will test the hypothesis that next generation sequencing will identify unique small RNAs in human prefrontal cortex and amygdala. This will provide definitive information about the presence and brain regional distribution of these small RNA regulatory molecules, which will not only be of general importance in neuroscience, but will advance a molecular classification of alcohol dependence in key regions of the brain. Aim 3 will test the hypothesis that next generation sequencing will reveal novel expression changes in functional groups of genes found in the prefrontal cortex and amygdala of human alcoholics and changes will be seen in homologous brain regions using mouse and macaque models of excessive alcohol consumption. To address similarities and differences in transcriptome changes found in animal models of excessive alcohol consumption and human alcoholics across INIA sites, we will utilize differential expression and Weighted Gene Covariance Network Analysis (WGCNA).

描述（由申请人提供）：我们假设长期的酗酒会导致大脑基因表达的变化，并且这些变化至少部分造成了酒精依赖性。我们还假设，特定基因的表达模式存在显着的结构变化，可以通过下一代测序检测到。该项目的目的是利用这种最先进的技术来确定在涉及良好特征的人类受试者的药物奖励中差异表达的新成绩单，这是酒精摄入量，滥用和依赖性的函数。迄今为止，在人类扩展的杏仁核和相关皮质投射区域中尚无公开的基因表达报告。这些研究的结果将确定尚未尚未达到的酒精反应性基因，包括低丰度，剪接变体和表达的SNP。与其他INIA调查人员合作，我们将有机会分析和比较酒精暴露后从人，猕猴和啮齿动物获得的序列数据。拟议的实验是授予PI的先前INIA West Pilot项目的直接结果。提出了三个具体目标：AIM 1将检验以下假设：与匹配的对照相比，下一代转录组（RNA-SEQ）将在特征良好的酒精依赖性病例的精选大脑区域中鉴定出差异表达的RNA。随后的RT-PCR分析将验证并定义哪些参与介导饮酒的影响。 AIM 2将测试下一代测序将在人前额叶皮层和杏仁核中识别独特的小RNA的假设。这将提供有关这些小RNA调节分子的存在和大脑区域分布的明确信息，这不仅在神经科学中至关重要，而且还将推进大脑关键区域中酒精依赖性的分子分类。 AIM 3将检验以下假设：下一代测序将揭示在前额叶皮层和人类酒精中毒的杏仁核中发现的官能基因的新表达变化，并且使用过量酒精消耗的小鼠和猕猴在同源的大脑区域中可以看到变化。为了解决非洲饮酒和人类酗酒的动物模型中发现的相似性和差异，我们将利用差异表达和加权基因协方差网络分析（WGCNA）。