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.

在没有药物暴露的情况下，可以通过以下方法在动物模型中评估风险作为一种群体指标：多种策略，例如通过使用选择性培育的动物品系。然而，建模风险并不是一个问题。即使在非人类模型中，过程也很简单，因为风险不是单一的结构，因此是多重遗传的；必须考虑非遗传因素，并且甲基苯丙胺的危险因素可能因人而异。 (MA) 具有强大的欣快作用，鼓励使用，但经过多年的研究，广泛有效；我们已经开发了一种由小鼠品系组成的遗传动物模型。针对高和低自愿 MA 饮酒（MAHDR 和 MALDR）进行选择性培育，目的是识别 MA 使用的遗传风险和保护因素这些品系因与 MA 密切相关的多个 MA 性状而异。我们绘制了小鼠 10 号染色体上的一个区域，该区域占遗传的 50% 以上。 MA消耗量的差异，并获得为微量胺相关受体提供证据的数据 1 基因 Taar1，作为 MA 摄入量的数量性状基因本应用重点关注 Taar1 与 MA 风险的关系。使用，以及发现与 Taar1 突变 (Taar1m1J) 相关的风险增加的遗传修饰剂编码非功能性受体；这将通过识别来完成。转录组的个体差异影响 Taar1m1J/m1J 基因型对 MA 摄入的影响。研究结果将指导治疗 MA 使用障碍的新机制的研究，其中三个目标是。在目标 1 中，CRISPR-Cas9 Taar1m1J 等位基因取代了小鼠，其中 TAAR1 功能已恢复。特别是在 MAHDR 小鼠 (MAHDR-Taar1+/+) 中，将与未替换的对照 (MAHDR-Taar1 m1J/m1J），可靠地区分 MAHDR 和 MALDR 所选品系（在 Taar1 中不同）的 MA 相关性状。基因型，将进行检查（即，MAHDR 均为 Taar1m1J/m1J，MALDR 均为 Taar1+/+ 或 Taar1+/m1J）。还将进行糖精（作为促味剂和自然奖励）和奎宁（作为促味剂）的行为。在目标 2 中，将使用 TAAR1 特异性激动剂来测试等位基因交换有效性的评估。 RNA-Seq 数据将用于重组近交系小鼠的基因表达网络分析 (BXD RI）均具有 Taar1m1J/m1J 基因型目标是识别减少影响的遗传修饰剂。 Taar1m1J/m1J 基因型对 MA 摄入量的影响，因为它们可能导致新的伏隔核治疗。 (NAc) 内侧壳因其在药物奖励中的关键作用而被首先研究。在目标 3 中，RNA-Seq 结果、数据。基础搜索（DrugBank、Broad Institute Connectivity Map、NCBI 的 PubChem）和发布的结果将用于指定神经目标来操纵它们对 MA 摄入的影响，并最终识别新的这项研究的长期目标是确定可以减少 MA 使用的新疗法。与由特定基因群组成的个体遗传易感性“标记”相匹配。