Bridging genetic variation with behavior: Molecular and functional mechanisms of quantitative trait gene regulation of the stimulant and addictive properties of methamphetamine in mice

将遗传变异与行为联系起来：数量性状基因调节小鼠甲基苯丙胺的兴奋和成瘾特性的分子和功能机制

• 批准号: 9926356
• 负责人: CAMRON D BRYANT
• 金额: $ 0.85万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9926356
• 关键词: ADRA1B gene; AGFG1 gene; Addictive Behavior; Adrenergic Agents; Alleles; Alternative Splicing; Animal Model; Behavior; Biological; Chromosomes, Human, Pair 11; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Congenic Mice; Corpus striatum structure; DARPP; Data; Disease; Down-Regulation; Early Endosome; Environmental Risk Factor; Etiology; Exhibits; FDA approved; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Polymorphism; Genetic Transcription; Genetic Variation; Genetic study; Genomics; Genotype; Glutamates; Heroin Dependence; Human; Human Genetics; Immunoblotting; Immunohistochemistry; Inherited; Intercistronic Region; Knock-in; Knock-in Mouse; Knock-out; Knockout Mice; Link; Measures; Mediating; Mental Health; Methamphetamine; Methamphetamine dependence; Microdialysis; Midbrain structure; Molecular; Motor Activity; Mus; Neurobiology; Neurons; Nucleotides; Oral; Pathway interactions; Patients; Phenotype; Predisposition; Prevention; Property; Proteins; Psychostimulant dependence; Public Health; Quantitative Trait Loci; RNA-Binding Proteins; Regulatory Element; Relapse; Research; Self Administration; Series; Signal Transduction; Signaling Molecule; Synaptic Transmission; System; Testing; Therapeutic; Transcription Coactivator; Translating; United States; Variant; Wild Type Mouse; addiction; congenic; differential expression; dopaminergic neuron; gene discovery; genetic variant; genome wide association study; glutamatergic signaling; in vivo; insight; mRNA sequencing; monoamine; mu opioid receptors; mutant; nerve supply; neurobiological mechanism; neurochemistry; neuron development; neurotransmission; neurotransmitter release; noradrenergic; novel; novel therapeutics; nuclease; null mutation; psychostimulant; public health relevance; receptor; resilience; response; targeted nucleases; tool; trait; transcription activator-like effector nucleases; transcriptome; transcriptome sequencing; transcriptomics; transmission process; violent crime

 DESCRIPTION (provided by applicant): Methamphetamine (MA) addiction is a pervasive public health concern in the U.S. and is associated with violent crime, mental health decline, and a high rate of relapse in recovering patients. Despite decades of research, there are still no FDA-approved treatments for MA addiction. Both genetic and environmental factors contribute to the etiology of MA addiction; however, genome-wide association studies in humans are limited in their power to identify common variants associated with this devastating disease. Mouse genetics offers a powerful, complementary tool for accelerating novel gene discovery and biological mechanisms. We recently identified a 210 kb region on mouse chromosome 11 containing two protein coding genes (Hnrnph1 and Rufy1) that contributes to the locomotor stimulant response to MA. The primary objective of this proposal is to identify the novel quantitative trait gene (QTG) and the neurobiological mechanism by which this QTG regulates the stimulant and addictive properties of MA. In Aim 1, we will identify the QTG(s) that regulates MA sensitivity. We generated a 112 kb congenic mouse containing only Hnrnph1 and Rufy1, providing the unique opportunity to determine whether this region is both necessary and sufficient for MA sensitivity. Furthermore, we recently used transcriptional activator-like effecto nucleases (TALENs) to introduce null mutations in Hnrnph1 and Rufy1. Preliminary studies provide strong, replicable evidence that Hnrnph1 is a QTG for reduced MA sensitivity. To fully tackle all of the polymorphisms within the 210 kb region that could potentially contribute to behavior, we will also delete a 150 kb intergenic region using the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system to introduce double stranded breaks that flank the intergenic region. Finally, we will use CRISPR/Cas9 to generate knockin mice carrying SNPs from the candidate QTG. In Aim 2, we will use transcriptome analysis of the striatum and ventral midbrain via mRNA sequencing (RNA- seq) in mice carrying the congenic allele, the knockout allele of the QTG, or both alleles relative to wild-type mice to hone the molecular pathways and neurobiological mechanisms that link genetic variation with behavior. Preliminary analysis of a congenic line capturing the QTL for reduced MA sensitivity suggests perturbations in midbrain dopaminergic neuron development, glutamate signaling, and adrenergic signaling as potentially responsible for a reduction in MA-induced behavior. In Aim 3, we will use in vivo microdialysis, immunoblotting, immunohistochemistry, and operant oral MA self-administration in congenics and knockouts to functionally assess the neurochemical mechanism by which the QTG influences the stimulant and addictive properties of MA. The proposed studies employ a unique combination of genetics, genomics, and functional analyses to uncover the neurobiological mechanism by which a novel genetic factor contributes to addiction-relevant behaviors. These findings will likely have translational relevance in the genetic and molecular mechanisms that confer susceptibility/resilience to MA addiction and could inform new therapeutic avenues.