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A Reduced Complexity Cross in BALB/c substrains to identify the genetic basis of oxycodone dependence phenotypes

BALB/c 亚种中降低复杂性的杂交以确定羟考酮依赖性表型的遗传基础

基本信息

批准号：
10673804
负责人：
CAMRON D BRYANT
金额：
$ 71万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10673804
关键词：
Adaptive Behaviors Address Affective Alleles Behavior Behavioral Biological Body Weight decreased Boston Brain region CRISPR/Cas technology Candidate Disease Gene Cells Cessation of life Chromosome Mapping Collaborations Collection Complement Control Locus Cues Dependence Dose Drug Addiction Emotional Epidemic Future Gene Expression Gene Expression Profiling Gene Expression Regulation Genes Genetic Genetic Crosses Genetic Variation Genetic study Genomics Genotype Health Hereditary Disease Heritability Human Human Genome Inbred BALB C Mice Inbred Mouse Learning Maps Memory Mental disorders Modeling Molecular Mus Neuroanatomy Neurobiology Neurons Nucleotides Opiate Addiction Opioid Opioid agonist Outcome Oxycodone Pathway interactions Pharmaceutical Preparations Phenotype Process Quantitative Trait Loci Resolution Retrieval Rewards Risk Speed Statistical Study Substance Use Disorder Time Tissue-Specific Gene Expression Tissues Training Treatment Protocols United States Universities Validation Variant Withdrawal Work addiction behavioral outcome brain tissue candidate validation causal variant cell type drug reward drug withdrawal gene discovery gene network genetic analysis genome wide association study glial activation improved in vivo innovation insight model organism motor learning mouse model mu opioid receptors neuroadaptation novel opioid withdrawal single nucleus RNA-sequencing trait transcriptome sequencing whole genome

项目摘要

PROJECT SUMMARY Substance use disorders (SUDs) are heritable psychiatric disorders with a significant genetic component. Opioid dependence, one of the most heritable SUDS, has reached epidemic proportions in the United States. Human genome-wide association studies (GWAS) are statistically underpowered to detect the majority of common genetic variation that contributes to opioid dependence. Discovery-based genetics in mammalian model organisms is a powerful complement to human GWAS and can uncover novel genetic factors, biological pathways, and gene networks underlying addiction traits. Mouse models are advantageous because they enable collection of the relevant brain tissue at the appropriate time points under controlled opioid dosing. Furthermore, gene editing permits the validation of functional variants in vivo within the same species on a controlled, genetic background. Reduced Complexity Crosses (RCCs) are genetic crosses between inbred mouse substrains that are nearly genetically identical and can vastly improve the speed at which causal genetic factors can be identified. Our primary objective is to use an RCC between BALB/c substrains to discover the genetic and molecular basis of opioid addiction-relevant traits at two stages of opioid dependence following repeated administration of the mu opioid receptor agonist oxycodone (OXY; the active ingredient of Oxycontin®). We found robust differences between BALB/c substrains in opioid adaptive behaviors, including state-dependent learning of OXY-induced locomotor stimulation and reward following limited, low-dose administration (1.25 mg/kg, IP) as well as the emotional-affective component of opioid withdrawal and weight loss following repeated high-dose administration (40 mg/kg, IP). In Aim 1, we will map quantitative trait loci (QTLs) underlying these OXY phenotypes in an RCC F2 cross. In Aim 2, we will map QTLs controlling gene expression (eQTLs) in the relevant brain tissues of control F2 mice and in OXY-trained F2 mice. We will then nominate candidate causal genes and nucleotides underlying behavior by integrating eQTL with behavioral QTL analysis. To increase precision in assigning candidate variants with the regulation of gene expression and behavior and to identify biological pathways and opioid-adaptive gene networks in specific cell types, we will use single nucleus RNA- seq (snRNA-seq) of brain tissue following limited, low-dose OXY and repeated high-dose OXY. In Aim 3, we will validate candidate functional variants underlying OXY phenotypes using CRISPR/Cas9 gene editing of each of the two alternate alleles onto each reciprocal substrain background. This approach will allow us to demonstrate both necessity and sufficiency of the quantitative trait nucleotides. The proposed studies will identify the genetic basis of unique opioid phenotypes across two stages of opioid dependence. Independent from gene discovery, these studies have broader application in revealing novel, actionable insight toward cellular adaptations at progressive stages of the opioid addiction process and potentially improving behavioral outcomes.

项目总结