Translatome profiling of nicotine addiction

尼古丁成瘾的翻译组分析

• 批准号: 9550950
• 负责人: Jubao Duan
• 金额: $ 23.7万
• 依托单位: NORTHSHORE UNIVERSITY HEALTHSYSTEM
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9550950
• 关键词: Addictive Behavior; Alleles; Amygdaloid structure; Biological Assay; Biological Process; Brain; Brain region; Candidate Disease Gene; Cause of Death; Chemicals; Chronic Disease; Data; Data Set; Development; Drug Addiction; Electrophysiology (science); Exhibits; Expression Profiling; Future; Gene Expression; Gene Targeting; Genes; Genetic; Genetic study; Harvest; Hippocampus (Brain); Human; Hybrids; Inbred Strain; Intravenous; Knowledge; Link; Maintenance; Mass Spectrum Analysis; Measures; Mediating; Messenger RNA; Midbrain structure; Modeling; Molecular; Nature; Neurobiology; Neurons; Nicotine; Nicotine Dependence; Noise; Norway; Nucleus Accumbens; Parents; Parkinson Disease; Pathologic; Pathway interactions; Pharmaceutical Preparations; Phase; Process; Property; Protein Biosynthesis; Proteins; Proteomics; RNA; Rat-1; Rattus; Rattus norvegicus; Resolution; Rewards; Ribosomes; Risk Factors; Rodent; Saline; Self Administration; Sex Characteristics; Smoker; Smoking; Smoking Behavior; Surrogate Markers; Testing; Therapeutic Intervention; Tobacco; Transcript; Translations; Treatment Efficacy; Variant; Ventral Tegmental Area; addiction; base; brain tissue; cigarette smoking; differential expression; dopaminergic neuron; genetic variant; genome wide association study; genome-wide; health economics; improved; induced pluripotent stem cell; innovation; insight; knowledge base; neurobiological mechanism; neurochemistry; nicotine abuse; nicotine exposure; nicotine use; non-smoker; novel; sex; trait; transcriptome; transcriptome sequencing; translatome; whole genome

Abstract Smoking is the foremost preventable cause of death. Nicotine (NIC) determines the addictive nature of smoking. In rats, NIC addition has been commonly modeled by intravenous NIC self-administration (SA). We (Dr. Vezina) have recently shown that NIC sensitization, a process that appears after repeated exposure to NIC, can enhance the SA of NIC and other stimulants. NIC sensitization and SA in rats are known to correlate with neurobiological changes in multiple brain regions, however, the molecular basis of the neurobiological changes of NIC sensitization and SA as well as its relevance to human NIC abuse has yet to be determined. We propose to take advantage of the depth and breadth of RNA sequencing (RNAseq) to identify the gene expression profiles of NIC sensitization and SA by quantifying transcripts actively involved in protein synthesis (i.e., translatome) in these processes, and to explore its relevance to enhanced NIC use in humans. Translatome profiling better predicts protein abundance than conventional transcriptome profiling and has higher resolution and dynamic range than mass spectrometry-based proteomic assays. To model NIC sensitization and SA, we will innovatively use F1 progeny of two inbred strains, Fischer-344 (F344) and Brown Norway (BN) since F1 rats are (1) genetically identical, thus minimizing “genetic noise” on measured differential expression and (2) heterozygous at all loci where parental strains have different alleles, thus maximizing the informative heterozygous variants for analyzing allele-specific expression (ASE). We have shown that these F1 rats (F344/BN) exhibit robust NIC sensitization. AIM1: We will characterize the translatome profiles of NIC sensitization and SA by sequencing RNA transcripts bound to ribosomes in ventral tegmental area (VTA) and nucleus accumbens (NAc), the brain regions most relevant to NIC addiction. We will further identify genetic variants showing ASE of sensitization-associated gene translation. AIM2: To examine the relevance of NIC addiction-associated translatome profiles to the enhanced NIC use in humans, we will derive induced pluripotent stem cells (iPSCs) and midbrain DA neurons from both smokers and non-smokers, expose DA neurons to NIC, and examine how the NIC-induced expression changes in iPSC-neurons correlate with those observed in rat brain tissues. For genes correlated with NIC sensitization, we will further evaluate their relevance to human smoking behavior by performing a pathway-based association test in publicly available smoking GWAS datasets. Identifying novel gene targets relevant to NIC sensitization and SA will forge a new path to deepen our understanding of the neurobiology of human NIC abuse, helping develop more effective therapeutic interventions.

抽象的 吸烟是可预防的死亡原因。尼古丁（NIC）确定 吸烟。在大鼠中，NIC添加通常是通过静脉注射自我给药（SA）建模的。我们 （Vezina博士）最近表明，NIC敏感性是在重复接触后出现的过程 NIC可以增强NIC和其他兴奋剂的SA。已知大鼠的NIC传感化和SA相关 然而，随着多个大脑区域的神经生物学变化，神经生物学的分子基础 NIC传感和SA的变化以及与人类NIC滥用的相关性尚未确定。 我们建议利用RNA测序的深度和广度（RNASEQ）来识别 NIC敏感性和SA的基因表达谱通过量化积极参与的转录本 这些过程中的蛋白质合成（即翻译组），并探索与增强的NIC相关性 用于人类。翻译成分比传统转录组更好地预测蛋白质抽象 分析比基于质谱的蛋白质组学测定法具有更高的分辨率和动态范围。到 Model NIC Sensorization和SA，我们将创新使用两种近交菌株Fischer-344（F344）的F1进度。 和棕色挪威（BN），因为F1大鼠是（1）遗传学相同，从而最大程度地减少了“遗传噪声” 测量的差异表达和（2）在父母菌株具有不同等位基因的所有地方的杂合子， 从而最大程度地提高了信息丰富的杂合变量，以分析等位基因特异性表达（ASE）。我们有 表明这些F1大鼠（F344/bn）暴露了可靠的NIC传感器。 AIM1：我们将表征 通过测序与腹侧核糖体结合的RNA转录本的Translatome Sensorization和SA的翻译组曲线 隔离区域（VTA）和伏隔核（NAC），大脑区域与NIC成瘾最相关。我们将 进一步鉴定遗传变异显示了敏感性相关基因翻译的ASE。 AIM2：检查 与NIC成瘾相关的翻译谱与增强的NIC在人类中使用的相关性，我们将 来自吸烟者和非吸烟者的诱导多能干细胞（IPSC）和中脑DA神经元， 将DA神经元暴露于NIC，并检查NIC诱导的IPSC-神经元中的表达如何变化 在大鼠脑组织中观察到的那些。对于与NIC传感器相关的基因，我们将进一步评估 通过公开进行基于途径的关联测试，它们与人类吸烟行为的相关性 可用的吸烟GWAS数据集。识别与NIC传感器相关的新型基因靶标和SA 为加深我们对人类NIC虐待神经生物学的理解加深了一条新的途径，帮助发展更多 有效的治疗干预措施。