Identifying genomic loci related to vulnerability to opioid addiction

识别与阿片类药物成瘾脆弱性相关的基因组位点

• 批准号: 10222637
• 负责人: JONATHAN GEWIRTZ
• 金额: $ 61.01万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10222637
• 关键词: ATAC-seq; Acute; Addictive Behavior; Anhedonia; Animal Genetics; Animal Model; Basic Science; Behavior; Behavioral; Bioinformatics; Biological; Biological Assay; ChIP-seq; Chromatin; Consumption; Cues; Data; Data Set; Drug usage; Economics; Epigenetic Process; Exhibits; Exploratory/Developmental Grant for Diagnostic Cancer Imaging; Exposure to; Extinction (Psychology); Future; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Genomics; Genotype; Goals; Impulsivity; Individual; Individual Differences; Intake; Maps; Measures; Medial; Minority; Modeling; Modification; Molecular Target; Morphine; National Institute of Drug Abuse; Opiate Addiction; Opioid; Pattern; Pharmaceutical Preparations; Phase; Phenotype; Play; Predisposition; Prefrontal Cortex; Prevention; Prevention approach; Prevention strategy; Rattus; Regulation; Regulator Genes; Relapse; Research; Rodent Model; Role; Self Administration; Severities; Sprague-Dawley Rats; System; Techniques; Transcriptional Activation; United States National Institutes of Health; Variant; Withdrawal; addiction; behavioral economics; behavioral phenotyping; bioinformatics tool; combinatorial; drug reinforcement; endophenotype; epigenetic regulation; epigenomics; experimental study; genetic variant; genomic data; genomic locus; improved; indexing; individual variation; innovation; next generation sequencing; novel; opioid use disorder; personalized approach; prospective; transcriptome sequencing; transcriptomics

Between 8-12% of people exposed to opioids develop opioid use disorder (OUD). Developing more effective preventions and treatments for OUD requires a better understanding of genomic and epigenetic mechanisms that underlie individual vulnerability to distinct stages along the OUD trajectory (e.g., initial use, compulsive use, relapse). The overall goal of this proposal is to use an outbred rat model (Sprague-Dawley) to identify novel downstream genes and upstream regulators of gene transcription involved in 3 behavioral phenotypes associated with distinct stages along the OUD trajectory. By comparing rats that show high versus low levels of addiction-like behavior at each stage, we will be able to identify changes in gene expression and their regulation associated specifically with susceptibility to opioid addiction from among the numerous effects of opioids that are unrelated to addiction. Anhedonia produced during withdrawal from acute morphine (i.e., withdrawal-induced anhedonia, WIA) will be used as an addiction phenotype of the earliest phase of OUD, i.e. prior to voluntary drug consumption. We have previously found that WIA is more strongly associated with a range of measures of subsequent i.v. morphine self-administration (SA) than these SA measures are with one another. Economic demand for morphine and reinstatement after extinction will serve as measures of drug reinforcement efficacy and propensity for relapse, respectively, after extensive morphine SA. To identify vulnerability-related genomic targets, we will use Next-Generation Sequencing (NGS) techniques and advanced bioinformatic tools to compare transcriptomic and epigenomic differences in rats exhibiting high versus low levels of WIA (Aim 1), demand (Aim 2) or reinstatement (Aim 3). Our epigenomic assays will map loci of chromosomal accessibility (using ATAC-seq) and of the stable chromatin mark H3K4me3 (using ChIP- seq). We will overlay each of these data sets onto RNA-seq data to identify genes showing differential activation in High- versus Low-Susceptibility rats, as well as upstream regulators of these transcriptional effects. These assays will focus on the medial prefrontal cortex (mPFC), a node within the mesocorticolimbic system that plays a pivotal role in addiction. We will also overlay epigenomic maps derived from our studies onto genotyping data derived from larger studies of Heterogeneous Stock (HS) rats manifesting High- versus Low addiction-related behavioral phenotypes (e.g., drug intake, impulsivity), in other NIDA Animal Genetics Consortium U01/P50 projects. We hypothesize that these comparisons will yield a set of downstream genes and upstream regulators associated with individual differences in early vulnerability to addiction-like behavior and its severity once established. We further hypothesize that our transcriptomic and epigenomic data will provide a viable roadmap for identifying genetic variants from larger genomic datasets associated with individual differences in OUD susceptibility. As such, our studies promise to yield novel genomic and molecular targets for developing more effective, individualized approaches for the prevention and treatment of OUD.

暴露于阿片类药物的人中，有8-12％的人患有阿片类药物使用障碍（OUD）。发展更有效 OUD的预防和治疗需要更好地了解基因组和表观遗传机制 这是个人沿OUD轨迹不同阶段的个人脆弱性的基础（例如，初始使用，强迫性 使用，复发）。该提案的总体目标是使用近代大鼠模型（Sprague-Dawley）来识别 新型的下游基因和基因转录的上游调节剂参与了3种行为表型 与OUD轨迹不同的阶段相关。通过比较显示高水平与低水平的大鼠 在每个阶段，类似成瘾的行为，我们将能够识别基因表达的变化及其它们的变化 与对阿片类药物成瘾易感性相关的调节，来自 与成瘾无关的阿片类药物。从急性吗啡退出期间产生的Anhedonia（即 戒断引起的Anhedonia，WIA）将用作最早的OUD阶段的成瘾表型，即 在自愿吸毒之前。我们以前已经发现WIA与 随后的i.v.的度量范围吗啡自我管理（SA）比这些SA措施与 其他。灭绝后吗啡和恢复原始的经济需求将作为药物的措施 广泛的吗啡SA之后，分别加强功效和复发的倾向。识别 与脆弱性相关的基因组目标，我们将使用下一代测序（NGS）技术和 先进的生物信息学工具比较表现出高的大鼠的转录组和表观基因组差异 与低水平的WIA（AIM 1），需求（AIM 2）或恢复原状（AIM 3）。我们的表观基因质分析将映射 染色体可及性的基因座（使用ATAC-SEQ）和稳定的染色质H3K4me3（使用芯片 - SEQ）。我们将这些数据集叠加到RNA-seq数据上，以识别显示差异的基因 高敏度大鼠的激活以及这些转录的上游调节剂 效果。这些测定将重点放在中内侧的前额叶皮层（MPFC）上，该节点是中质质体内的节点 在成瘾中起关键作用的系统。我们还将覆盖从我们的研究中得出的表观基因组图 从表现高的异质库存（HS）大鼠的较大研究中得出的基因分型数据 其他NIDA动物遗传学 联盟U01/p50项目。我们假设这些比较将产生一组下游基因 以及与早期脆弱性类似成瘾行为的个体差异相关的上游调节器 它的严重性曾经建立。我们进一步假设我们的转录组和表观基因组数据将 提供可行的路线图，以识别与与之相关的较大基因组数据集的遗传变异 OUD敏感性的个体差异。因此，我们的研究有望产生新的基因组和分子 开发更有效，个性化方法的目标，以预防和治疗OUD。