Regulation of Gene Enhancers in Human Heroin Use

项目总结/摘要-项目4 项目4的目标是描述基因组增强子区域的影响， 对基因表达的关键控制，在鸦片成瘾中。对几种组织的研究表明， 增强子区域由几种特定的组蛋白修饰“标记”，例如H3 K27 ac（组蛋白3Lys 27 乙酰化），并且富含几种染色质重塑因子。一个例子是BRD 4， BET（布罗莫结构域和额外末端）亚家族，其通过其 溴结构域，并在几种组织中与增强子紧密连接。我们发现组织中 在患有海洛因使用障碍的人的纹状体中H3 K27 ac和BRD 4两者， 与多年海洛因使用和关键基因网络的表达水平密切相关。类似的监管也出现在 自我注射海洛因的老鼠的纹状体。此外，我们发现， BRD 4/BET功能-用于癌症治疗的临床开发-在海洛因中发挥治疗样作用 啮齿动物的成瘾模型。本项目将以几种方式促进这些研究。我们将获得基因组- 海洛因使用障碍患者和匹配对照的NAc和DS中增强子区域的广泛图谱， 以及在海洛因自我给药后的大鼠中，通过使用ChIP-seq检测H3 K27 ac和BRD 4等标记， 基因和染色质分析核心。这将在分离的细胞核上单独进行 这些脑区的神经元和非神经元细胞。我们将与核心合作， 覆盖这些数据集的生物信息学方法，这是增强子的可靠鉴定所需的， 沿着我们目前在相同样本上生成的RNA-seq和ATAC-seq数据集， 将提供进一步了解特定的靶基因受确定的海洛因调节增强子。 我们将通过研究双向调节的作用来检验增强子调节在海洛因成瘾中的因果作用。 NAc和DS中BRD 4对大鼠模型中基因表达和行为终点的操纵，包括 筛选几种BRD 4/BET抑制剂的抗成瘾作用。这些研究提供了一个翻译 这是一个推进药物成瘾新疗法的机会。

PROJECT SUMMARY/ABSTRACT– PROJECT 4 Project 4’s objective is to characterize the influence of enhancer regions of the genome, which exert crucial control over gene expression, in opiate addiction. Work in several tissues has demonstrated that enhancer regions are “marked” by several specific histone modifications, such as H3K27ac (histone 3 Lys27 acetylation), and are enriched in several chromatin remodeling factors. One example is BRD4, a member of the BET (bromodomain and extra-terminal) subfamily, which binds acetylated histones through its bromodomains and is tightly linked with enhancers in several tissues. We have found elevated tissue levels of both H3K27ac and BRD4 in striatum of humans with heroin use disorders, with such elevations correlating strongly with years of heroin use and expression levels of key gene networks. Similar regulation is seen in striatum of rats that self-administer heroin. Moreover, we have found that small molecules that disrupt BRD4/BET function—in clinical development for cancer treatment—exert therapeutic-like effects in heroin addiction models in rodents. This Project will further these studies in several ways. We will obtain genome- wide maps of enhancer regions in NAc and DS of humans with heroin use disorders and matched controls, and in rats after heroin self-administration, by use of ChIP-seq for H3K27ac and BRD4, among other marks, in conjunction with the Gene and Chromatin Analysis Core. This will be performed separately on nuclei isolated from neurons and non-neuronal cells of these brain regions. We will work with the Core on advanced bioinformatics approaches to overlay these datasets, which is required for reliable identification of enhancers, along with RNA-seq and ATAC-seq datasets that we are currently generating on the same samples and that will provide further insight into the specific target genes affected by the identified heroin-regulated enhancers. We will test the causal role of enhancer regulation in heroin addiction by studying the effect of bidirectional manipulations of BRD4 in NAc and DS on gene expression and behavioral endpoints in rat models, including screening several BRD4/BET inhibitors for anti-addiction actions. These studies offer a translational opportunity to advance novel therapies for drug addiction.