Chan Siu Chiu, Selth Luke A, Li Yingming, Nyquist Michael D, Miao Lu, Bradner James E, Raj Ganesh V, Tilley Wayne D, Dehm Scott M
Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55905, USA.
Dame Roma Mitchell Cancer Research Laboratories and Freemasons Foundation Centre for Mens' Health, School of Medicine, The University of Adelaide, Adelaide, SA 5005, Australia.
Nucleic Acids Res. 2015 Jul 13;43(12):5880-97. doi: 10.1093/nar/gkv262. Epub 2015 Apr 23.
Androgen receptor (AR) variants (AR-Vs) expressed in prostate cancer (PCa) lack the AR ligand binding domain (LBD) and function as constitutively active transcription factors. AR-V expression in patient tissues or circulating tumor cells is associated with resistance to AR-targeting endocrine therapies and poor outcomes. Here, we investigated the mechanisms governing chromatin binding of AR-Vs with the goal of identifying therapeutic vulnerabilities. By chromatin immunoprecipitation and sequencing (ChIP-seq) and complementary biochemical experiments, we show that AR-Vs display a binding preference for the same canonical high-affinity androgen response elements (AREs) that are preferentially engaged by AR, albeit with lower affinity. Dimerization was an absolute requirement for constitutive AR-V DNA binding and transcriptional activation. Treatment with the bromodomain and extraterminal (BET) inhibitor JQ1 resulted in inhibition of AR-V chromatin binding and impaired AR-V driven PCa cell growth in vitro and in vivo. Importantly, this was associated with a novel JQ1 action of down-regulating AR-V transcript and protein expression. Overall, this study demonstrates that AR-Vs broadly restore AR chromatin binding events that are otherwise suppressed during endocrine therapy, and provides pre-clinical rationale for BET inhibition as a strategy for inhibiting expression and chromatin binding of AR-Vs in PCa.
在前列腺癌(PCa)中表达的雄激素受体(AR)变体(AR-Vs)缺乏AR配体结合域(LBD),并作为组成型活性转录因子发挥作用。患者组织或循环肿瘤细胞中的AR-V表达与对AR靶向内分泌治疗的抗性及不良预后相关。在此,我们研究了控制AR-Vs染色质结合的机制,目的是确定治疗弱点。通过染色质免疫沉淀和测序(ChIP-seq)及补充性生化实验,我们表明AR-Vs对与AR优先结合的相同经典高亲和力雄激素反应元件(AREs)表现出结合偏好,尽管亲和力较低。二聚化是组成型AR-V DNA结合和转录激活的绝对必要条件。用溴结构域和额外末端(BET)抑制剂JQ1处理导致AR-V染色质结合受到抑制,并损害了AR-V驱动的PCa细胞在体外和体内的生长。重要的是,这与下调AR-V转录本和蛋白质表达的新型JQ1作用相关。总体而言,本研究表明AR-Vs广泛恢复了在内分泌治疗期间否则会被抑制的AR染色质结合事件,并为BET抑制作为抑制PCa中AR-Vs表达和染色质结合的策略提供了临床前理论依据。