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发展的研究领域。与 与乙酰化组蛋白结合并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)家族(BRD2、BRD3、BRD4和BRDT)具有 成为很有希望的治疗靶点。然而，到目前为止，只有PAN-BET溴域抑制剂，小 与所有BET蛋白中的两个溴结构域(Bd1和BD2)结合的分子已经在SUD- 相关实验。在SUD模型中使用这些非选择性药物抑制物限制了我们的 对单个BET蛋白、溴结构域选择功能(BD1vs.BD2)和非BET的机制的理解 调节药物诱导的神经行为适应的溴域BET相互作用。来自我们的新证据 实验室表明，选择性地靶向单个BET结构域和蛋白质是一种新颖而有效的方法 减少可卡因诱导的行为和转录反应而不产生副作用的策略 用PAN-BET抑制剂观察。以这些令人兴奋的数据为基础，推进表观遗传学领域 对于sud的药物治疗，我们建议使用高度选择性的、临床相关的治疗方法，由病毒介导。 研究BET蛋白结构域和蛋白质特异性机制的方法和多组学分析 在寻找可卡因的行为中。为了实现这些目标，我们将首先调查行为和细胞类型- 可卡因经济需求和可卡因需求中区域选择性BET抑制剂的特异性转录反应 复职程序。接下来，我们将使用免疫共沉淀和质谱分析来表征 短期和间歇使用可卡因后BRD4相互作用组的变化 研究中，我们将使用新开发的工具来确定BRD4的非溴域机制在 可卡因自我管理。最后，我们将询问BRD2在转录和行为方面的新角色 寻找可卡因的行为。总而言之，这些实验将为 成瘾、表观遗传学以及减少可卡因使用和复发的新治疗目标。

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.