1] The Breakthrough Breast Cancer Research Centre and CRUK Gene Function Laboratory, Institute of Cancer Research, London, UK [2] Département de médecine-Unité INSERM 981, Institut Gustave Roussy, Villejuif, France.
Oncogene. 2013 Nov 21;32(47):5377-87. doi: 10.1038/onc.2013.311. Epub 2013 Aug 12.
Excision repair cross-complementation group 1 (ERCC1) is a DNA repair enzyme that is frequently defective in non-small cell lung cancer (NSCLC). Although low ERCC1 expression correlates with platinum sensitivity, the clinical effectiveness of platinum therapy is limited, highlighting the need for alternative treatment strategies. To discover new mechanism-based therapeutic strategies for ERCC1-defective tumours, we performed high-throughput drug screens in an isogenic NSCLC model of ERCC1 deficiency and dissected the mechanism underlying ERCC1-selective effects by studying molecular biomarkers of tumour cell response. The high-throughput screens identified multiple clinical poly (ADP-ribose) polymerase 1 and 2 (PARP1/2) inhibitors, such as olaparib (AZD-2281), niraparib (MK-4827) and BMN 673, as being selective for ERCC1 deficiency. We observed that ERCC1-deficient cells displayed a significant delay in double-strand break repair associated with a profound and prolonged G₂/M arrest following PARP1/2 inhibitor treatment. Importantly, we found that ERCC1 isoform 202, which has recently been shown to mediate platinum sensitivity, also modulated PARP1/2 sensitivity. A PARP1/2 inhibitor-synthetic lethal siRNA screen revealed that ERCC1 deficiency was epistatic with homologous recombination deficiency. However, ERCC1-deficient cells did not display a defect in RAD51 foci formation, suggesting that ERCC1 might be required to process PARP1/2 inhibitor-induced DNA lesions before DNA strand invasion. PARP1 silencing restored PARP1/2 inhibitor resistance in ERCC1-deficient cells but had no effect in ERCC1-proficient cells, supporting the hypothesis that PARP1 might be required for the ERCC1 selectivity of PARP1/2 inhibitors. This study suggests that PARP1/2 inhibitors as a monotherapy could represent a novel therapeutic strategy for NSCLC patients with ERCC1-deficient tumours.
切除修复交叉互补基因 1(ERCC1)是一种 DNA 修复酶,在非小细胞肺癌(NSCLC)中经常存在缺陷。尽管 ERCC1 表达水平低与铂类药物敏感性相关,但铂类药物治疗的临床效果有限,这突显了需要替代治疗策略。为了发现针对 ERCC1 缺陷肿瘤的新机制为基础的治疗策略,我们在 ERCC1 缺陷的同源 NSCLC 模型中进行了高通量药物筛选,并通过研究肿瘤细胞反应的分子生物标志物来剖析 ERCC1 选择性作用的机制。高通量筛选确定了多种临床多聚(ADP-核糖)聚合酶 1 和 2(PARP1/2)抑制剂,如奥拉帕利(AZD-2281)、尼拉帕利(MK-4827)和 BMN 673,对 ERCC1 缺陷具有选择性。我们观察到,与 PARP1/2 抑制剂治疗后明显和持久的 G₂/M 期阻滞相关,ERCC1 缺陷细胞的双链断裂修复明显延迟。重要的是,我们发现最近已显示出介导铂类药物敏感性的 ERCC1 同工型 202 也调节了 PARP1/2 敏感性。PARP1/2 抑制剂合成致死性 siRNA 筛选表明,ERCC1 缺陷与同源重组缺陷呈上位性。然而,ERCC1 缺陷细胞未显示 RAD51 焦点形成缺陷,表明 ERCC1 可能需要在 DNA 链入侵之前处理 PARP1/2 抑制剂诱导的 DNA 损伤。PARP1 沉默恢复了 ERCC1 缺陷细胞对 PARP1/2 抑制剂的耐药性,但对 ERCC1 功能正常的细胞没有影响,支持 PARP1 可能是 PARP1/2 抑制剂对 ERCC1 选择性所必需的假设。这项研究表明,PARP1/2 抑制剂作为单一疗法可能代表 ERCC1 缺陷肿瘤的 NSCLC 患者的一种新的治疗策略。