School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, 510515, China.
Department of Pharmacy, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China.
Acta Pharmacol Sin. 2022 Jul;43(7):1803-1815. doi: 10.1038/s41401-021-00783-5. Epub 2021 Nov 4.
The tumor suppressor gene BAP1 encodes a widely expressed deubiquitinase for histone H2A. Both hereditary and acquired mutations are associated with multiple cancer types, including cutaneous melanoma (CM), uveal melanoma (UM), and clear cell renal cell carcinoma (ccRCC). However, there is no personalized therapy for BAP1-mutant cancers. Here, we describe an epigenetic drug library screening to identify small molecules that exert selective cytotoxicity against BAP1 knockout CM cells over their isogenic parental cells. Hit characterization reveals that BAP1 loss renders cells more vulnerable to bromodomain and extraterminal (BET) inhibitor-induced transcriptional alterations, G1/G0 cell cycle arrest and apoptosis. The association of BAP1 loss with sensitivity to BET inhibitors is observed in multiple BAP1-deficient cancer cell lines generated by gene editing or derived from patient tumors as well as immunodeficient xenograft and immunocompetent allograft murine models. We demonstrate that BAP1 deubiquitinase activity reduces sensitivity to BET inhibitors. Concordantly, ectopic expression of RING1A or RING1B (H2AK119 E3 ubiquitin ligases) enhances sensitivity to BET inhibitors. The mechanistic study shows that the BET inhibitor OTX015 exerts a more potent suppressive effect on the transcription of various proliferation-related genes, especially MYC, in BAP1 knockout cells than in their isogenic parental cells, primarily by targeting BRD4. Furthermore, ectopic expression of Myc rescues the BET inhibitor-sensitizing effect induced by BAP1 loss. Our study reveals new approaches to specifically suppress BAP1-deficient cancers, including CM, UM, and ccRCC.
肿瘤抑制基因 BAP1 编码一种广泛表达的组蛋白 H2A 去泛素化酶。遗传性和获得性突变与多种癌症类型相关,包括皮肤黑色素瘤(CM)、葡萄膜黑色素瘤(UM)和透明细胞肾细胞癌(ccRCC)。然而,目前针对 BAP1 突变型癌症尚无个性化治疗方法。在这里,我们描述了一种针对表观遗传药物库的筛选,以鉴定对 BAP1 敲除 CM 细胞具有选择性细胞毒性的小分子,而对其同基因亲本细胞则没有。命中特征描述表明,BAP1 缺失使细胞对溴结构域和末端(BET)抑制剂诱导的转录改变、G1/G0 细胞周期停滞和细胞凋亡更加敏感。在通过基因编辑产生的多个 BAP1 缺陷型癌细胞系以及源自患者肿瘤的细胞系以及免疫缺陷性异种移植和免疫活性同种异体移植小鼠模型中,观察到 BAP1 缺失与对 BET 抑制剂的敏感性之间存在关联。我们证明 BAP1 去泛素化酶活性降低了对 BET 抑制剂的敏感性。一致地,RING1A 或 RING1B(H2AK119 E3 泛素连接酶)的异位表达增强了对 BET 抑制剂的敏感性。机制研究表明,BET 抑制剂 OTX015 在 BAP1 缺失细胞中的转录抑制作用比在其同基因亲本细胞中更强,对各种增殖相关基因(尤其是 MYC)的转录抑制作用更强,主要通过靶向 BRD4。此外,Myc 的异位表达挽救了由 BAP1 缺失引起的 BET 抑制剂增敏作用。我们的研究揭示了专门抑制 BAP1 缺陷型癌症(包括 CM、UM 和 ccRCC)的新方法。