Elucidating causal mechanisms of ethanol-induced analgesia in BXD recombinant inbred mouse lines

阐明 BXD 重组近交系小鼠乙醇诱导镇痛的因果机制

• 批准号: 10825737
• 负责人: Walker David Rogers
• 金额: $ 4.21万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-10 至 2026-09-09
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10825737
• 关键词: Absence of pain sensation; Acute; Address; Affect; Alcohol consumption; Alcohols; Algorithms; American; Analgesics; Animals; Behavioral; Behavioral Genetics; Bioinformatics; Biological; Biological Assay; Brain region; Candidate Disease Gene; Characteristics; Data; Databases; Development; Dose; Ensure; Environmental Risk Factor; Ethanol; Event; GIRK2 subunit, G protein-coupled inwardly-rectifying potassium channel; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Gene Modified; Gene Targeting; Genes; Genetic; Genetic study; Genotype; Heritability; Human; Hyperalgesia; Inbred Mouse; Informatics; Knockout Mice; Link; Literature; Maps; Mediation; Methodology; Molecular; Mus; Nociception; Opioid Receptor; Pain; Pain management; Pathway Analysis; Pathway interactions; Pattern; Peripheral Nervous System Diseases; Persons; Phenotype; Population; Potassium Channel; Prefrontal Cortex; Process; Property; Public Health; Quantitative Trait Loci; Reaction Time; Recombinants; Reporting; Research; Risk; Rodent Model; Role; Sampling; Signal Pathway; Source; Symptoms; System; Tissue-Specific Gene Expression; Training; United States; Variant; Viral Vector; Withdrawal; addiction; alcohol use disorder; antinociception; attenuation; behavioral phenotyping; behavioral response; bioinformatics resource; candidate identification; candidate validation; chronic alcohol ingestion; chronic pain; chronic painful condition; design; diagnostic criteria; differential expression; experience; experimental study; gene network; genetic architecture; genome wide association study; genome-wide; genomic locus; large-scale database; mechanical allodynia; midbrain central gray substance; model organism; mouse genome; mouse model; neural circuit; overexpression; pain symptom; phenomics; response; training opportunity; trait; transcriptome sequencing; transcriptomics; trend; whole genome

Project Summary Alcohol use disorder (AUD) and pain are major public health problems affecting millions of people in the United States. More than 14 million Americans met the diagnostic criteria for AUD in 2019, and 50 million people in the United States were estimated to have a chronic pain condition. These public health crises are related, as people experiencing pain often report using alcohol to manage pain symptoms, and people with chronic pain are at an elevated risk for developing AUD. Drinking alcohol to alleviate pain is a maladaptive strategy and may lead to withdrawal-induced hyperalgesia, peripheral neuropathy, and AUD. While alcohol confers analgesic effects in a dose-dependent manner, genetic differences mean that not every person experiences this analgesia to the same degree. Previous research has revealed moderate-to-strong heritability (h2=0.42) for ethanol-induced analgesia in C57BL/6J and DBA/2J mice, but the genes underlying this relationship are still poorly characterized. Studies in rodent models have revealed evidence that opioid receptor systems, GIRK2 potassium channels, and GABAergic signaling pathways all have roles in ethanol analgesia, but no whole- genome interrogations have been conducted. This project aims to identify candidate genes modulating alcohol- induced analgesia using complementary genetic, behavioral and transcriptomic experiments in genetically informative BXD mouse populations. By mapping quantitative trait loci (QTL) for both basal and post-ethanol hot plate latency with BXD strains, we will identify genomic loci linked with variation in ethanol-induced analgesia. As an additional genome-wide approach, prefrontal cortex and periaqueductal gray samples from high- and low-ethanol analgesia BXD strains will be studied by RNA-seq. We will perform weighted gene co- expression network analysis (WGCNA) to identify differentially expressed genes in these two analgesia groups. By combining evidence from behavioral, statistical, and molecular methodologies, we will select the strongest ethanol analgesia candidate gene for experimental verification using either overexpression or knockout mouse models. This proposal comprises an investigative progression from identifying candidate genes by QTL mapping, to elucidating gene expression patterns in critical brain regions, to validating ethanol analgesia candidate genes. It represents an outstanding training opportunity for the candidate to design and perform a suite of systems genetics and animal behavioral experiments and contribute to the scientific understanding of substance-induced analgesia.

项目摘要 酒精使用障碍（AUD）和疼痛是影响美国数百万人的主要公共卫生问题。 States. 2019年，超过1400万美国人符合AUD的诊断标准， 据估计，美国人患有慢性疼痛症。这些公共卫生危机是相互关联的， 经历疼痛的人经常报告使用酒精来控制疼痛症状， 发展为AUD的风险较高。喝酒来缓解疼痛是一种适应不良的策略， 可能导致戒断诱发的痛觉过敏、周围神经病变和AUD。虽然酒精赋予 镇痛作用的剂量依赖性，遗传差异意味着不是每个人的经验， 这种镇痛作用。先前的研究表明，对于遗传性疾病，具有中度到强的遗传力（h2=0.42）。 C57 BL/6 J和DBA/2 J小鼠中乙醇诱导的镇痛，但这种关系的基因仍然存在。 特征不佳。在啮齿动物模型中的研究显示，阿片受体系统，GIRK 2 钾通道和GABA能信号通路都在乙醇镇痛中发挥作用，但没有整体- 进行了基因组研究。该项目旨在确定调节酒精的候选基因- 使用互补的遗传、行为和转录组实验在遗传学上诱导镇痛 信息丰富的BXD小鼠群体。通过定位基础和乙醇后的数量性状位点（QTL）， 热板潜伏期与BXD菌株，我们将确定基因座与乙醇诱导的变异， analgesia.作为一种额外的全基因组方法，来自大脑皮层的前额叶皮层和中脑导水管周围灰质样本 将通过RNA-seq研究高和低乙醇镇痛BXD菌株。我们将进行加权基因共- 表达网络分析（WGCNA），以确定差异表达的基因在这两个镇痛 组通过结合行为学、统计学和分子方法学的证据，我们将选择 最强的乙醇镇痛候选基因，用于使用过表达或 敲除小鼠模型。该提案包括从确定候选人 通过QTL定位基因，阐明关键脑区的基因表达模式，验证乙醇 镇痛候选基因它为候选人提供了一个出色的培训机会， 执行一套系统遗传学和动物行为实验，并有助于科学 了解物质诱导的镇痛。