Domain- and protein-selective BET mechanisms in cocaine-seeking behaviors

项目摘要 物质使用障碍（SUD）的表观遗传药物治疗是一个迅速扩大的研究领域。与 结合乙酰化组蛋白和调节药物诱导的转录适应的能力， 溴结构域和末端外结构域（BET）蛋白家族（BRD 2、BRD 3、BRD 4和BRDT）具有 成为有前途的治疗靶点。然而，迄今为止，只有泛BET布罗莫结构域抑制剂，小的 结合所有BET蛋白内的两个溴结构域（BD 1和BD 2）的分子，已经在SUD中进行了测试。 相关实验。在SUD模型中使用这些非选择性药理学抑制剂限制了我们的研究。 对单个BET蛋白、溴结构域选择性功能（BD 1与BD 2）和非溴结构域选择性功能的机制理解 溴结构域BET相互作用，调节药物诱导的神经行为适应。我们的新证据 实验室表明，选择性靶向个别BET结构域和蛋白质是一种新的和有效的 减少可卡因诱导的行为和转录反应而不引起副作用的策略 用泛BET抑制剂观察。以这些令人兴奋的数据为基础， 药物治疗SUD，我们建议使用高度选择性，临床相关的治疗，病毒介导的 方法和多组学分析来研究BET蛋白的结构域和蛋白质特异性机制 在寻求可卡因的行为中。为了实现这些目标，我们将首先研究行为和细胞类型- 结构域选择性BET抑制剂在可卡因经济需求中的特异性转录组学反应， 恢复程序。接下来，使用免疫共沉淀和质谱，我们将表征 短距离和快速接触可卡因自我给药后BRD 4相互作用组的变化， 研究，我们将使用新开发的工具来确定BRD 4的非布罗莫结构域机制在 可卡因自我管理最后，我们将询问BRD 2在细胞中的新的转录和行为作用。 寻求可卡因的行为总之，这些实验将为该领域做出重大贡献。 成瘾表观遗传学以及新的治疗目标，以减少可卡因的使用和复发。

Project Summary Epigenetic pharmacotherapy for substance use disorder (SUD) is a rapidly expanding area of research. With the ability to bind to acetylated histones and regulate drug-induced transcriptional adaptations, members of the bromodomain and extra-terminal domain (BET) family of proteins (BRD2, BRD3, BRD4 and BRDT) have emerged as promising therapeutic targets. To date, however, only pan-BET bromodomain inhibitors, small molecules that bind to both bromodomains (BD1 and BD2) within all BET proteins, have been tested in SUD- related experiments. The use of these non-selective pharmacological inhibitors in SUD models limits our mechanistic understanding of individual BET proteins, bromodomain-selective functions (BD1 vs. BD2), and non- bromodomain BET interactions that regulate drug-induced neurobehavioral adaptations. New evidence from our laboratory indicates that selectively targeting individual BET domains and proteins is a novel and effective strategy to reduce cocaine-induced behavioral and transcriptional responses without causing sides effects observed with pan-BET inhibitors. To build on these exciting data and to advance the field of epigenetic pharmacotherapy for SUD, we propose to use highly selective, clinically relevant treatments, viral-mediated approaches, and multiomic analysis to investigate domain- and protein-specific mechanisms of BET proteins during cocaine-seeking behaviors. To achieve these goals, we will first investigate the behavioral and cell type- specific transcriptomic responses of domain-selective BET inhibitors in cocaine economic demand and reinstatement procedures. Next, using co-immunoprecipitation and mass-spectrometry, we will characterize BRD4 interactome changes following short- and intermittent-access cocaine self-administration, and in functional studies, we will use newly developed tools to identify a role for non-bromodomain mechanisms of BRD4 in cocaine self-administration. Finally, we will interrogate novel transcriptional and behavioral roles for BRD2 in cocaine-seeking behaviors. Together, these experiments will provide significant contributions to the field of addiction epigenetics as well as novel therapeutic targets to reduce cocaine use and relapse.