Exploiting zebrafish genetics to identify genes affecting addiction-related phenotypes.

利用斑马鱼遗传学来识别影响成瘾相关表型的基因。

基本信息

批准号：
9751829
负责人：
Caroline Helen Brennan
金额：
$ 42.82万
依托单位：
UNIV/LONDON-QUEEN MARY& WESTFIELD COLL
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9751829
关键词：
Adult Affect Alcohols Alleles Animal Model Behavior Behavioral Behavioral Assay Behavioral Genetics Biological Factors Biological Process Biology Breeding CRISPR/Cas technology Candidate Disease Gene Cardiovascular Diseases Cause of Death Cell physiology Cells Cessation of life Chronic Data Development Developmental Gene Disease Drug Addiction Epidemic Evolution Family Fishes Future Gene Expression Gene Expression Profiling Generations Genes Genetic Genetic Models Genetic Risk Genetic Screening Genetic Variation Genome Genotype Goals Health Care Costs Heritability Human Human Genetics Human Genome Impulsivity Individual Knock-out Malignant Neoplasms Maps Mediating Mental disorders Microarray Analysis Modeling Mutagenesis Mutation Nicotine Nicotine Dependence Outcome Outcome Study Pathway interactions Phenotype Population Predisposition Reaction Time Regulator Genes Research Rewards Risk Siblings Smoking Substance abuse problem System Testing Time Tobacco Tobacco Dependence Variant Vertebrates Work World Health Organization Zebrafish addiction causal variant cost density developmental genetics drug of abuse drug reward exome sequencing gene function genome wide association study improved individualized medicine insight mutant neural circuit neural patterning neuronal patterning next generation sequencing nicotine exposure novel personalized medicine population based preference programs protein function response screening smoking addiction transcriptome sequencing treatment strategy

项目摘要

Exploiting zebrafish genetics to identify genes affecting addiction-related phenotypes The cost of substance abuse in the US alone is estimated at $166 billion annually in preventable health care costs, with tobacco ($130 billion) and alcohol ($25 billion) abuse contributing to the overwhelming majority of this expense. The World Health Organization declared tobacco-related disease a global epidemic, predicted to cause an estimated 8 million annual deaths worldwide by 2030, if unabated. There is a vital need for increased understanding of the genetic and cell biological factors influencing vulnerability to addiction including smoking to help identify individuals at risk and to inform treatment strategies. As approaches to identify genetic risk are difficult in humans, research on genetic risk can be facilitated by the use of animal models. Zebrafish are an established developmental and behavioral genetic model species that show conserved responses to common drugs of abuse as well as behaviors that model core phenotypes predictive of vulnerability to addiction. We shall identify genes affecting core phenotypes associated with addiction by screening lines of ethyl nitrosurea (ENU)-mutagenised zebrafish for sensitivity to nicotine reward and impulsivity. We test the hypothesis that genes that show persistent adaptive changes in expression following chronic nicotine exposure are also addiction vulnerability genes. Our specific aims are: 1) To isolate 25 heritable loci affecting a) nicotine reward and b) impulsivity. i) We will use behavioral assays in adult ENU-mutagenized zebrafish to identify families showing altered nicotine reward and impulse control. We have developed a scalable, fully automated behavioral assay system for this purpose. We will use analysis of microarray or RNAseq data to identify genes that show persistent changes in expression following chronic nicotine exposure as candidates for genes affecting nicotine addiction vulnerability. We will pre-load the screen with families carrying mutations in these candidate genes. ii) We shall use exome sequencing to map the mutations in at least 10 families. The use of ENU- mutagenized lines facilitates identification of the causal mutations (by generation of strong phenotypes and/or by the ENU mutations acting as markers). 2) To identify the cell biological processes underlying the observed behavioral phenotypes. i) We shall use additional (predicted functional) ENU mutations or targeted knockouts of the identified genes to confirm genotype-phenotype associations. ii) We shall perform RNA sequencing to gain insight into possible effects on gene expression and cellular pathways affected. iii) We shall perform developmental analyses to test the hypothesis that behavioral effects are mediated by effects on neuronal patterning or neural circuit formation at developmental timepoints. The expected outcomes from this study are identification of novel genetic factors influencing core behaviors predictive of vulnerability to drug addiction and increased understanding of the cellular processes affected. These findings will enable future human genetic variation studies in these identified genes, with the potential to develop more personalized therapies. Results from this work will have a positive impact on individualized treatment efforts for tobacco dependence and addiction as a whole.

利用斑马鱼遗传学来识别影响成瘾相关表型的基因仅在美国，就可以预防健康，仅在美国滥用药物的费用为1660亿美元护理费用，烟草（1,300亿美元）和酒精（250亿美元）的滥用这笔费用的大部分。世界卫生组织宣布与烟草有关的疾病全球流行病预测，如果没有减弱，到2030年，全世界估计将导致大约800万例死亡。至关重要的需要增加对影响遗传和细胞生物学因素的理解成瘾的脆弱性，包括吸烟，以帮助识别有风险的人并告知治疗策略。由于人类难以识别遗传风险的方法，因此对遗传风险的研究可能是通过使用动物模型来促进。斑马鱼是既定的发展和行为遗传模型物种显示出对常见滥用药物以及行为的保守反应该模型核心表型可以预测成瘾的脆弱性。我们将确定影响核心的基因通过筛选硝基尿素（ENU） - 淡核化的筛选线与成瘾相关的表型斑马鱼对尼古丁奖励和冲动的敏感性。我们检验了显示的基因的假设慢性尼古丁暴露后表达的持续自适应变化也是成瘾脆弱性基因。我们的具体目的是： 1）分离25个可遗传的基因座影响a）尼古丁奖励和b）冲动。 i）我们将在成人核糖核酸斑马鱼中使用行为分析来识别家庭显示改变尼古丁的奖励和冲动控制。我们已经开发了一个可扩展的，充分的为此目的，自动行为测定系统。我们将使用微阵列的分析或RNASEQ数据以识别显示表达式持续变化以下的基因慢性尼古丁暴露是影响尼古丁成瘾基因的候选者脆弱性。我们将在其中携带突变的家庭预装屏幕候选基因。 ii）我们将使用外显子组测序来绘制至少10个家庭的突变。使用诱变线促进了因果突变的识别（通过产生强烈的表型和/或通过充当标记的ENU突变）。 2）确定观察到的行为表型的基础的细胞生物学过程。 i）我们将使用其他（预测的功能性）ENU突变或针对性的敲除确定的基因确认基因型 - 表型关联。 ii）我们将进行RNA测序以深入了解对基因的可能影响表达和细胞途径受到影响。 iii）我们将进行发育分析以检验行为效应的假设通过对神经元模式或神经回路形成的影响介导的发展时间点。这项研究的预期结果是对影响核心的新遗传因素的鉴定可预测吸毒成瘾的脆弱性和对细胞的理解增加的行为流程受影响。这些发现将使未来的人类遗传变异研究在这些发现基因，有可能开发更多个性化疗法。这项工作的结果将有一个对整个烟草依赖性和成瘾的个性化治疗工作的积极影响。