Gene Regulation in the Opioid Dependent Human Brain (Project 2)

阿片类药物依赖性人脑的基因调控（项目 2）

基本信息

批准号：
10493706
负责人：
Peter Christopher Scacheri
金额：
$ 58.91万
依托单位：
RESEARCH TRIANGLE INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10493706
关键词：
Acute Addictive Behavior Address Amygdaloid structure Architecture Autopsy Basal Ganglia Behavior Behavioral Biological Brain Brain region Cells ChIP-seq Chronic Collection Complex DNA Methylation Data Data Analyses Data Set Dependence Development Dimensions Dorsal Environmental Risk Factor Fluorescence-Activated Cell Sorting Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Genetic Engineering Genome Genomics Genotype Goals Histone Acetylation Human Impulsivity Individual Injections Institutes Intervention Intoxication Lateral Link Maps Measurement Meta-Analysis Methylation Motivation Mus Neurobiology Nucleus Accumbens Opiate Addiction Opioid Pathway interactions Pharmacology Prefrontal Cortex Public Health Quantitative Trait Loci RNA Regulator Genes Regulatory Element Resources Rewards SET gene Sampling Single Nucleotide Polymorphism Site Stress Substance Use Disorder Substance abuse problem Surveys Testing Time Tissues Transcript Validation Variant Viral Vector Withdrawal Work addiction brain tissue case control cell type cohort compulsion drug discovery epigenome executive function follow-up genetic association genetic variant genome wide association study genome-wide gun homicide histone modification human tissue in vivo incentive salience multiple omics negative affect novel novel therapeutics opioid abuse opioid epidemic opioid exposure opioid mortality opioid overdose opioid use overdose death relating to nervous system single cell analysis single-cell RNA sequencing synergism targeted treatment theories transcriptome transcriptome sequencing

项目摘要

PROJECT SUMMARY/ABSTRACT The ongoing opioid epidemic is one of the most important public health crises of our time with overdose fatalities quadrupling since 1999 and surpassing those of gun homicides in recent years. Given the individual and societal burden of opioid dependence, it is necessary to characterize the biological pathways to dependence that can be targeted for treatment and intervention. The work proposed in Project 2 combines multi-omics resources to examine specific hypotheses regarding the functional brain architecture of opioid addiction (OA). Guided by existing theory implicating the prefrontal cortex (PFC), nucleus accumbens (NAc), and amygdala in the cycle of substance abuse, we propose to generate and combine existing data at multiple omics levels in each brain region using postmortem human tissue collected from opioid overdose cases and matched controls. The measurement approaches we use are genome-wide, in each of the three brain regions and in each biological domain: single nucleotide polymorphism (SNP), epigenome (DNA methylation and histone acetylation), and transcript (via RNA-seq) in our combined sample (N=641) with follow-up single-cell validation/replication. We will accomplish these goals across three aims. In Aim 1, in the existing combined sample, we will fill out the brain regions not currently assessed in specific brain/regions or biological domains and perform analyses to detect region-specific and general (across-brain regions) methylation, histone acetylation, and gene expression differences between cases and controls. In addition, we will perform analyses aimed at integrating across those biological domains. In Aim 2, we will perform single-cell follow-up of Aim 1 findings to pinpoint all cell types within the PFC, NAc, and amygdala that drive the differences identified in Aim 1, with validation by RNA-seq in fluorescence activated cell sorting (FACS) separated cells and RNA-FISH. In Aim 3, we will genotype samples where needed and perform genetic association mapping—histone modification, DNA methylation, and gene expression QTL mapping—to identify genetic variants that account for the regulatory differences observed between cases and controls in Aim 1. These QTL approaches will allow us to identify regulatory differences with a genetic basis, versus those somatically acquired as a consequence of chronic opiate use, exposure, or other environmental factors. Our overall goal is to create a framework in which we can map, in brain, the OA functional relevance of genomic sets (genes, pathways, features) and generate results that can be leveraged in Project 1 and the Synergy Core through mapOA, and in Project 3 through selected genes, targeted for functional validation in vivo in the dlPFC and NAc using genetic engineering, viral vector injection, and pharmacology approaches in mice. Through this synergy we will distinguish between predisposing and exposure consequent dysregulation and move toward mechanistic understanding that can drive new treatments of OA.

项目概要/摘要持续的阿片类药物流行是我们这个时代最重要的公共卫生危机之一，因为药物过量自 1999 年以来，死亡人数翻了两番，并且近年来超过了枪支杀人的死亡人数。和阿片类药物依赖的社会负担，有必要描述其生物学途径项目 2 中提出的工作将治疗和干预作为目标。多组学资源来检查有关阿片类药物大脑功能结构的具体假设以涉及前额皮质（PFC）、伏隔核（NAc）的现有理论为指导。和杏仁核在药物滥用循环中的作用，我们建议生成并合并现有数据使用从阿片类药物过量中收集的死后人体组织，在每个大脑区域进行多个组学水平我们在每个病例和匹配的对照中使用的测量方法是全基因组的。三个大脑区域和每个生物领域：单核苷酸多态性（SNP）、表观基因组（DNA 甲基化和组蛋白乙酰化），以及我们的组合样本（N=641）中的转录物（通过RNA-seq）后续单细胞验证/复制。我们将通过三个目标来实现这些目标，在目标 1 中，我们将在现有的组合样本中进行填充。排除当前未在特定大脑/区域或生物领域评估的大脑区域，并执行分析以检测区域特异性和一般（跨脑区域）甲基化、组蛋白乙酰化和此外，我们将针对病例和对照之间的基因表达差异进行分析。在目标 2 中，我们将对目标 1 的发现进行单细胞随访查明 PFC、NAc 和杏仁核内驱动目标 1 中确定的差异的所有细胞类型，通过荧光激活细胞分选 (FACS) 分离细胞和 RNA-FISH 中的 RNA-seq 进行验证。 3、我们将在需要的地方对样本进行基因分型并进行遗传关联图谱——组蛋白修饰， DNA 甲基化和基因表达 QTL 作图——识别导致基因突变的遗传变异在目标 1 中观察到病例和对照之间的监管差异。这些 QTL 方法将使我们能够识别具有遗传基础的调节差异，与因身体获得的调节差异长期使用阿片类药物、接触阿片类药物或其他环境因素。我们的总体目标是创建一个框架，在该框架中我们可以在大脑中映射 OA 功能相关性基因组集（基因、途径、特征）并生成可在项目 1 和 Synergy Core 通过 mapOA，并在项目 3 中通过选定的基因，旨在进行功能验证使用基因工程、病毒载体注射和药理学方法在 dlPFC 和 NAc 中进行体内研究通过这种协同作用，我们将区分诱发因素和暴露后果。调节失调并朝着可推动 OA 新疗法的机制理解迈进。