Heroin-Induced genomic regulation of Ventral Pallidum neuron subtypes

海洛因诱导的腹侧苍白球神经元亚型的基因组调控

• 批准号: 10210378
• 负责人: Seth Abrams Ament
• 金额: $ 66.76万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 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/10210378
• 关键词: ATAC-seq; Abstinence; Acute; Animal Model; Applications Grants; BRAIN initiative; Back; Behavior; Behavioral; Biological Assay; Brain; CRISPR interference; Cell Nucleus; Cells; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Cues; Data; Data Set; Dependence; Dependovirus; Dorsal; Drug Addiction; Epidemic; Epigenetic Process; Foundations; Frequencies; Functional disorder; Funding; Gene Expression; Genes; Genetic; Genetic Transcription; Genomics; Globus Pallidus; Goals; Habenula; Health; Heroin; Heroin Dependence; Hour; Individual; Knowledge; Label; Lateral; Lead; Life; Link; Medial; Mediating; Messenger RNA; Modeling; Molecular; Neurobiology; Neurons; Neurosciences; Nucleus Accumbens; Opiate Addiction; Opioid; Output; Overdose; Pharmaceutical Preparations; Process; Proteins; Rattus; Regulation; Regulator Genes; Relapse; Research; Research Personnel; Rewards; Role; Self Administration; Thalamic structure; Therapeutic; Time; Tissues; Training; Transcription Process; Ventral Tegmental Area; addiction; cell type; chromatin immunoprecipitation; combat; dopaminergic neuron; drug abstinence; drug craving; drug seeking behavior; epigenome; epigenomics; falls; genome-wide; genomic locus; genomic tools; insight; molecular subtypes; multimodal data; multiple omics; nerve supply; network models; novel; opioid abuse; opioid use; opioid use disorder; psychostimulant; tool; transcriptome; transcriptome sequencing; transcriptomics; web portal

PROJECT SUMMARY/ABSTRACT Opioid use, dependence, and addiction have dramatically increased to epidemic proportions in recent years, leading to substantial financial and societal health burdens, as well as an increasing number of overdoses. To combat this epidemic, it is imperative that we understand the neurobiological underpinnings that lead to opioid use disorder. We must identify disrupted neuron subtypes in the brain in opioid use disorders and dysregulated molecules within these neurons that underlie cellular, circuit, and ultimately behavioral adaptations. Use of rat drug self-administration (SA) and relapse assays, which are considered the best available animal models of addiction, will allow a more complete understanding of the molecular mechanisms underlying the genomic, epigenetic, and transcriptional-induced cellular plasticity that drives the long-lasting drug seeking and propensity for heroin relapse. We will perform genome-wide transcriptome and open-chromatin profiling of ventral pallidum (VP) projection neuron subtypes in rat heroin SA, both acutely following drug cessation and after prolonged periods of drug abstinence. Here, we focus on the VP as a critical node in the brain’s reward circuit. Our studies will profile VP neurons that project to the nucleus accumbens, ventral tegmental area, medial dorsal thalamus, and lateral habenula. We will then integrate the transcriptomic and epigenomic data with complementary transcriptomic and epigenomic datasets, including multimodal data from the BRAIN Initiative describing cell type diversity in the VP and its output circuits. We will reconstruct cell type-specific gene co-expression and open chromatin networks and identify hub genes predicted to have central roles in immediate and prolonged abstinence from heroin, which could underlie subsequent relapse behavior. This collection of datasets and models will be made available through a biologist-friendly web portal based on our BRAIN Initiative-funded Neuroscience Multi-Omic Analytics platform. Using the data generated we will develop rat gene loci-specific CRISPR epigenomic targeting tools to determine the functional significance of key hub genes that are regulated in VP projection neuron subtypes. To achieve this goal, we will employ rat models of relapse in combination with advanced CRISPRa and CRISPRi AAV tools to enhance or reduce transcription of key hub genes during heroin SA or abstinence from heroin SA followed by cue-induced reinstatement. The studies proposed in this grant application will, for the first time, identify the distinct heroin-induced gene network adaptations occurring temporally in a cell-type-specific manner within a novel neurobiological circuit.

项目摘要/摘要 近年来，阿片类药物的使用，依赖性和成瘾对流行比例的增加显着增加 导致大量的金融和社会健康伯恩根（Burnens），以及越来越多的过量剂量。到 战斗这种流行病，我们必须了解导致阿片类药物的神经生物学基础 使用障碍。我们必须在阿片类药物使用障碍中确定大脑中的神经元亚型的破坏和失调 这些神经元内的分子，这些神经元是细胞，回路和最终行为适应的基础。使用大鼠 药物自我管理（SA）和继电器测定法，它们被认为是最好的动物模型 成瘾，将使对基因组基础的分子机制有更完整的了解， 表观遗传学和转录诱导的细胞可塑性，驱动持久的药物寻求和承诺 用于海洛因接力。 我们将执行全基因组转录组和腹侧颗粒（VP）投影的开染色质分析 大鼠海洛因SA中的神经元亚型，既急性戒烟，又是在延长药物后 节制。在这里，我们将重点放在副总裁作为大脑奖励电路中的关键节点。我们的研究将介绍VP 投射到伏隔核，腹侧侧侧区域，内侧丘脑的神经元，然后 Habenula。然后，我们将将转录组和表观基因组数据与完整的转录组和 表观基因组数据集，包括来自大脑倡议的多模式数据，描述了VP中的细胞类型多样性 及其输出电路。我们将重建细胞类型特异性基因共表达和开放染色质网络 并确定被预测的中心基因在直接和长期戒酒中具有核心作用，而海洛因的戒酒是 可能是随后的救济行为的基础。将提供此数据集和模型的集合 通过基于我们大脑倡议资助的神经科学多摩变分析的生物学家友好的Web门户网站 平台。 使用生成的数据，我们将开发大鼠基因基因座特异性CRISPPR表观基因组靶向工具来确定 在VP投影神经元亚型中调节的关键集线器基因的功能意义。实现这一目标 目标，我们将使用大鼠退休模式与先进的CRISPRA和CRISPRI AAV工具结合使用 增强或减少海洛因SA期间关键枢纽基因的转录或海洛因SA的禁欲，然后是 提示引起的恢复。本赠款申请中提出的研究将首次确定 不同的海洛因诱导的基因网络适应暂时以细胞类型的方式发生 新型神经生物学回路。