Identification of gene variants mediating the behavioral and physiological response to THC

鉴定介导 THC 行为和生理反应的基因变异

• 批准号: 10660808
• 负责人: Bob Mack Moore
• 金额: $ 75.8万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10660808
• 关键词: Absence of pain sensation; Accounting; Acute; Address; Alleles; American; Attitude; Behavior; Behavioral; Biological; Brain; Brain region; CNR1 gene; Candidate Disease Gene; Cannabinoids; Cannabis; Chromosome Mapping; Complex; Consumption; Corpus striatum structure; Data; Dose; Exposure to; Female; Genes; Genetic; Genetic Engineering; Genetic Variation; Genomics; Genotype; Goals; Health; Heritability; Human; Hybrids; Inbred Mouse; Individual; Individual Differences; Knowledge; Liquid Chromatography; Locomotion; Maps; Mediating; Membrane; Mental Depression; Motor; Mus; Neurons; Outcome; Parents; Pharmacogenomics; Phenotype; Physiological; Policies; Population; Proteins; Proteome; Proteomics; Psychotropic Drugs; Public Policy; Quantitative Trait Loci; Receptor Activation; Receptor Signaling; Recombinants; Regulation; Risk; Rodent; Role; Sex Differences; Signal Pathway; Signal Transduction; Spectrometry; THC exposure; Testing; Tetrahydrocannabinol; Tissues; United States; Validation; Variant; Work; antinociception; candidate validation; cannabinoid receptor; cohort; differential expression; genetic variant; genomic locus; mRNA Expression; male; marijuana use; natural hypothermia; neurotransmission; pharmacologic; receptor; regional difference; response; sex; trafficking; trait; transcriptome; transcriptome sequencing; virtual

Cannabis is one of the most widely used psychotropic drugs in the United States and changes in public policy are expected to further increase use. Current cultivars of cannabis have been selected for high Δ9- tetrahydrocannabinol (THC) levels and THC levels have tripled over the past few decades. There is increasing evidence that use of high potency cannabis containing high levels of THC increases the risk of adverse health and behavioral consequences. Many of the specific effectors (e.g., receptors, receptor signaling pathways) mediating response to THC have not yet been fully identified and virtually nothing is known about how individual genetic differences influence response. This lack of knowledge represents a critical barrier in understanding health risks and behaviors associated with high potency cannabis use. To address these issues we have developed a powerful pharmacogenomic screen in the BXD recombinant inbred mouse population to identify gene variants in effector proteins that regulate cannabinoid 1 receptor (CB1) levels, signal transduction, termination of signaling, and/or trafficking following acute exposure to high-dose (10 mg/kg, i.p.) THC. Previously, we demonstrated strain differences in the level of the major effector of THC responses, CB1, between BXD parental strains and demonstrated heritable variation in acute responses to THC in parental strains and BXDs. We have already identified suggestive quantitative trait loci (QTLs) and candidate causal QTL genes (QTGs) in our pharmacogenomic screen using only a subset (i.e., 20) of BXDs. Here, we expand our work to identify genetic modulators of CB1 protein levels (Aim 1) and to identify and validate genetic modulators of acute responses to high-dose THC (Aim 2). In Aim 1A we use a targeted proteomics approach to quantify striatal CB1 levels in membrane-enriched protein fractions in parents, reciprocal F1, and BXD strains. Differential expression in parental strains, allele-specific expression in the F1 and linkage mapping in the BXDs will identify and validate QTLs containing trans-acting regulators of striatal CB1. In Aim 1B we extend the analysis to two other regions and in Aim 1C we include exposure to high-dose THC. In Aim 2A we screen a large panel (i.e., 80) of BXDs for differential CB1 activation quantified as hypolocomotion, hypothermia, and antinociception following VEH or THC treatment and identify QTLs accounting for at least 30% of the variation in each trait at a target precision of ±2 Mb. In Aim 2B we generate matched proteomes and transcriptomes for parental strains and F1 and generate transcriptome data for expression QTL (eQTL) mapping in BXDs with the goal of facilitating the identification of causal QTGs. In Aim 2C we validate the biological role of QTGs in cannabinoid receptor signaling pathways and responses to THC. This is the first pharmacogenomic screen to identify variation in effectors of CB1 signaling and will reveal new targets underlying responses to high-dose THC and behaviors associated with cannabis use in rodents and humans.

大麻是美国使用最广泛的精神药物之一，公共政策的变化 期望进一步增加使用。为高δ9-选择了当前的大麻品种 在过去的几十年中，四氢大麻酚（THC）水平和THC水平增加了两倍。有在增加 使用高效力大麻（含有高水平的四氢大麻酚）的证据增加了不良健康的风险 和行为后果。许多特定效应（例如受体，受体信号通路） 介导对THC的反应尚未得到充分识别，几乎没有什么了解 个体遗传差异会影响反应。缺乏知识代表了一个关键的障碍 了解与高效力大麻使用相关的健康风险和行为。解决这些问题 我们已经在BXD重组近交小鼠种群中开发了强大的药物基因组学筛查 鉴定调节大麻素1受体（CB1）水平的效应蛋白中的基因变异，信号 急性暴露于高剂量（10 mg/kg，i.p.）后的转导，信号传导的终止和/或运输 THC。以前，我们证明了THC响应CB1，CB1的主要效应因子水平的应变差异 在BXD的父母菌株之间表现出对父母中THC的急性反应的可遗传变化 菌株和BXD。我们已经确定了暗示性的定量性状区域（QTL）和候选催化 我们的药物基因组筛查中的QTL基因（QTG）仅使用BXD的子集（即20）。在这里，我们扩展 我们识别CB1蛋白水平的遗传调节剂（目标1）并识别和验证遗传的工作 对大剂量THC的急性反应的调节剂（AIM 2）。在AIM 1A中，我们使用靶向蛋白质组学方法 量化父母，倒数F1和BXD中膜含有膜含量蛋白质的纹状体CB1水平 菌株。亲本菌株中的差异表达，等位基因在F1中的特定表达和链接映射 BXD将识别并验证含有纹状体CB1的跨作用调节剂的QTL。在AIM 1B中 将分析扩展到其他两个区域，在AIM 1C中，我们包括暴露于大剂量THC。在AIM 2A中我们 筛选一个大型面板（即80），用于差异CB1激活的BXDS（80），被定量为降压运动， 体温过低和抗伤害感受在VEH或THC治疗后，并识别至少考虑的QTL 目标精度为±2 Mb的每个性状的30％变化。在AIM 2B中，我们生成匹配的蛋白质组织 和父母菌株和F1的转录组，并生成表达式QTL（EQTL）的转录组数据 在BXD中映射，目的是支持识别因果QTG。在AIM 2C中，我们验证 QTG在大麻素受体信号通路和对THC的反应中的生物学作用。这是第一个 药物基因组学筛查以识别CB1信号效应的变化，并将揭示新目标 对啮齿动物和人类中使用大麻使用的大剂量THC和行为的基本反应。