Departments of Medical Oncology, Radiation Oncology, and Cancer Biology, Dana-Farber Cancer Institute and Harvard Medical School, Boston, MA 02215.
Proc Natl Acad Sci U S A. 2013 Oct 15;110(42):17041-6. doi: 10.1073/pnas.1305170110. Epub 2013 Oct 1.
Breast Cancer Type 1 Susceptibility Protein (BRCA1)-deficient cells have compromised DNA repair and are sensitive to poly(ADP-ribose) polymerase (PARP) inhibitors. Despite initial responses, the development of resistance limits clinical efficacy. Mutations in the BRCA C-terminal (BRCT) domain of BRCA1 frequently create protein products unable to fold that are subject to protease-mediated degradation. Here, we show HSP90-mediated stabilization of a BRCT domain mutant BRCA1 protein under PARP inhibitor selection pressure. The stabilized mutant BRCA1 protein interacted with PALB2-BRCA2-RAD51, was essential for RAD51 focus formation, and conferred PARP inhibitor as well as cisplatin resistance. Treatment of resistant cells with the HSP90 inhibitor 17-dimethylaminoethylamino-17-demethoxygeldanamycin reduced mutant BRCA1 protein levels and restored their sensitivity to PARP inhibition. Resistant cells also acquired a TP53BP1 mutation that facilitated DNA end resection in the absence of a BRCA1 protein capable of binding CtIP. Finally, concomitant increased mutant BRCA1 and decreased 53BP1 protein expression occur in clinical samples of BRCA1-mutated recurrent ovarian carcinomas that have developed resistance to platinum. These results provide evidence for a two-event mechanism by which BRCA1-mutant tumors acquire anticancer therapy resistance.
BRCA1 缺陷型细胞的 DNA 修复能力受损,对聚 ADP 核糖聚合酶(PARP)抑制剂敏感。尽管最初有反应,但耐药的发展限制了临床疗效。BRCA1 的 BRCA C 末端(BRCT)结构域的突变经常导致无法折叠的蛋白质产物,这些产物易受蛋白酶介导的降解。在这里,我们在 PARP 抑制剂选择压力下显示 HSP90 介导的 BRCA1 蛋白 BRCT 结构域突变体的稳定。稳定的突变 BRCA1 蛋白与 PALB2-BRCA2-RAD51 相互作用,对 RAD51 焦点形成至关重要,并赋予 PARP 抑制剂和顺铂耐药性。用 HSP90 抑制剂 17-二甲基氨基乙基氨基-17-脱甲氧基格尔德霉素处理耐药细胞,降低了突变 BRCA1 蛋白水平,并恢复了对 PARP 抑制的敏感性。耐药细胞还获得了 TP53BP1 突变,从而在没有能够与 CtIP 结合的 BRCA1 蛋白的情况下促进 DNA 末端切除。最后,在对铂类药物耐药的 BRCA1 突变复发性卵巢癌的临床样本中,同时出现突变 BRCA1 增加和 53BP1 蛋白表达减少。这些结果为 BRCA1 突变肿瘤获得抗癌治疗耐药性的两事件机制提供了证据。