Genetic Factors Underlying Risk for Methamphetamine Intake and Associated Traits

甲基苯丙胺摄入风险和相关特征的遗传因素

基本信息

批准号：
10215457
负责人：
TAMARA J. RICHARDS
金额：
$ 34.7万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
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：
2018-09-30 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10215457
关键词：
Affect Agonist Alleles Amines Animal Genetics Animal Model Animals Behavior Behavior assessment Behavioral Breeding CRISPR/Cas technology Candidate Disease Gene Chromosome 10 Code Complex Confidence Intervals Consumption Crime Cyclic AMP Data Data Collection Development Disease Dopamine Drug Controls Drug Exposure Effectiveness Exhibits Gene Expression Gene Expression Profile Generations Genes Genetic Genetic Predisposition to Disease Genetic Variation Genomics Genotype Goals Homozygote Human Inbred Strain Inbred Strains Mice Individual Individual Differences Institutes Intake Knock-in Knock-in Mouse Lead Maps Measures Medial Methamphetamine Methamphetamine dependence Modeling Mus Mutation Nucleus Accumbens Oral Other Genetics Outcome Pathway Analysis Pharmaceutical Preparations Population Process Psychological reinforcement PubChem Publishing Quantitative Trait Loci Quinine Recombinant Inbred Strain Recording of previous events Research Rewards Risk Risk Factors Role Saccharin Self Administration Single Nucleotide Polymorphism Testing Tissues Translating Variant Violence addiction attenuation base child neglect conditioned place preference drinking drug reward effectiveness evaluation genetic manipulation genetic risk factor individual variation methamphetamine effect methamphetamine use monoamine multidrug abuse mutant non-genetic novel predictive test progenitor prospective protective factors receptor relating to nervous system response searchable database trait transcriptome transcriptome sequencing treatment response

项目摘要

Risk as a population measure can be assessed in animal models in the absence of drug exposure through a number of strategies, such as via the use of selectively bred animal lines. However, modelling risk is not a straightforward process, even in non-human models, since risk is not a unitary construct; thus, multiple genetic and non-genetic factors must be considered, and risk factors likely vary across individuals. Methamphetamine (MA) has powerful euphoric effects that encourage use; but, after many years of research, broadly effective medications have not been identified. We have developed a genetic animal model comprised of lines of mice selectively bred for high and low voluntary MA drinking (MAHDR and MALDR), with the goal of identifying genetic risk and protective factors for MA use. These lines differ for multiple MA traits critically relevant to MA use disorders. We mapped a region on mouse chromosome 10, that accounts for >50% of the genetic variance in MA consumption, and obtained data that provide evidence for the trace amine-associated receptor 1 gene, Taar1, as a quantitative trait gene for MA intake. This application is focused on Taar1 in risk for MA use, and on the discovery of genetic modifiers of the increased risk associated with a Taar1 mutation (Taar1m1J) that codes for a non-functional receptor; TAAR1. This will be accomplished through the identification of individual differences in the transcriptome that impact the effect of the Taar1m1J/m1J genotype on MA intake. Our findings will guide the examination of novel mechanisms for the treatment of MA use disorders. Three aims are proposed. In Aim 1, CRISPR-Cas9 Taar1m1J allele replaced mice, in which TAAR1 function has been restored specifically in MAHDR mice (MAHDR-Taar1+/+) will be compared to non-replaced controls (MAHDR-Taar1 m1J/m1J). MA-related traits that reliably differentiate the MAHDR and MALDR selected lines, which differ in Taar1 genotype, will be examined (i.e., MAHDR are all Taar1m1J/m1J and MALDR are Taar1+/+ or Taar1+/m1J). Studies with saccharin (as a tastant and a natural reward) and quinine (as a tastant) will also be performed. Behavioral assessment of the effectiveness of the allele swap will be tested using a TAAR1-specific agonist. In Aim 2, RNA-Seq data will be used in gene expression network analyses in recombinant inbred strains of mice (BXD RI) that all possess the Taar1m1J/m1J genotype. The goal is to identify genetic modifiers that reduce the impact of the Taar1m1J/m1J genotype on MA intake, because they could lead to new treatments. The nucleus accumbens (NAc) medial shell will be initially studied due for its critical role in drug reward. In Aim 3, RNA-Seq results, data base searches (DrugBank, Broad Institute Connectivity Map, PubChem at NCBI) and published results will be used to nominate neural targets to manipulate for their impact on MA intake and ultimately to identify novel treatments. The long-term goal of this research is to identify novel treatments that could reduce MA use, based on matching to individual genetic susceptibility “markers”, comprised of specific constellations of genes.

在没有通过A的药物暴露的情况下，可以在动物模型中评估风险作为人口措施数量的策略，例如通过使用有选择的繁殖动物线。但是，建模风险不是即使在非人类模型中，由于风险不是统一的结构，因此即使在非人类模型中也很简单。因此，多个通用必须考虑非遗传因素，风险因素可能因个人而异。甲基苯丙胺（MA）具有强大的欣快效果，可以鼓励使用；但是，经过多年的研究，有效尚未确定药物。我们已经开发了一个由小鼠系组成的遗传动物模型有选择地为高自愿和低自愿饮酒（Mahdr和Maldr）繁殖，目的是识别 MA使用的遗传风险和受保护因素。这些线与多种特征与MA有关的多种特征有所不同使用疾病。我们在鼠标10上绘制了一个区域，该区域占通用的50％ MA消耗的差异，并获得了提供痕量胺相关接收器证据的数据 1基因，taar1，作为MA摄入的定量性状基因。该应用的重点是有MA风险的TAAR1 在发现与TAAR1突变相关的风险增加（TAAR1M1J）的遗传修饰符时这代码为非功能接收器； taar1。这将通过确定影响TAAR1M1J/M1J基因型对MA摄入的影响的转录组的个体差异。我们的调查结果将指导检查新机制以治疗MA使用障碍。三个目标是建议的。在AIM 1中，CRISPR-CAS9 TAAR1M1J等位基因替换了TAAR1功能已恢复的老鼠将MAHDR小鼠（MAHDR-TAAR1+/+）中的特定于非替代对照（mahdr-taar1）进行比较（MAHDR-TAAR1） M1J/M1J）。与MA相关的特征可靠地区分MAHDR和MALDR选定的线，在TAAR1中有所不同基因型将进行检查（即Mahdr都是TAAR1M1J/M1J，MALDR为TAAR1+/+或TAAR1+/M1J）。研究还将执行糖精（作为味觉和自然奖励）和奎宁（作为味觉）。行为评估等位基因交换的有效性将使用TAAR1特异性激动剂进行测试。在AIM 2中， RNA-seq数据将用于小鼠重组近交菌株的基因表达网络分析（BXD） RI）所有都具有TAAR1M1J/M1J基因型。目的是确定减少影响的遗传修饰符 TAAR1M1J/M1J在MA摄入量上的基因型，因为它们可能导致新的治疗方法。伏隔核（NAC）内侧壳最初将因其在药物奖励中的关键作用而造成研究。在AIM 3中，RNA-seq结果，数据基本搜索（药品银行，广泛研究所的连接图，NCBI的PubChem）和已发布的结果将为用于提名神经目标以操纵其对MA摄入的影响，并最终确定新颖治疗。这项研究的长期目标是确定可以减少MA使用的新型治疗方法与单个遗传易感性“标记”匹配，该易感性“标记”由基因的特定星座组成。