Danish Cancer Society Research Center, Copenhagen, Denmark.
Cell Cycle. 2012 Oct 15;11(20):3837-50. doi: 10.4161/cc.22026. Epub 2012 Sep 14.
Impaired DNA damage response pathways may create vulnerabilities of cancer cells that can be exploited therapeutically. One such selective vulnerability is the sensitivity of BRCA1- or BRCA2-defective tumors (hence defective in DNA repair by homologous recombination, HR) to inhibitors of the poly(ADP-ribose) polymerase-1 (PARP-1), an enzyme critical for repair pathways alternative to HR. While promising, treatment with PARP-1 inhibitors (PARP-1i) faces some hurdles, including (1) acquired resistance, (2) search for other sensitizing, non-BRCA1/2 cancer defects and (3) lack of biomarkers to predict response to PARP-1i. Here we addressed these issues using PARP-1i on 20 human cell lines from carcinomas of the breast, prostate, colon, pancreas and ovary. Aberrations of the Mre11-Rad50-Nbs1 (MRN) complex sensitized cancer cells to PARP-1i, while p53 status was less predictive, even in response to PARP-1i combinations with camptothecin or ionizing radiation. Furthermore, monitoring PARsylation and Rad51 foci formation as surrogate markers for PARP activity and HR, respectively, supported their candidacy for biomarkers of PARP-1i responses. As to resistance mechanisms, we confirmed the role of the multidrug resistance efflux transporters and its reversibility. More importantly, we demonstrated that shRNA lentivirus-mediated depletion of 53BP1 in human BRCA1-mutant breast cancer cells increased their resistance to PARP-1i. Given the preferential loss of 53BP1 in BRCA-defective and triple-negative breast carcinomas, our findings warrant assessment of 53BP1 among candidate predictive biomarkers of response to PARPi. Overall, this study helps characterize genetic and functional determinants of cellular responses to PARP-1i and contributes to the search for biomarkers to exploit PARP inhibitors in cancer therapy.
DNA 损伤反应途径受损可能会使癌细胞产生易损性,从而可以进行治疗性利用。一种这样的选择性易损性是 BRCA1 或 BRCA2 缺陷肿瘤(因此在同源重组(HR)的 DNA 修复中存在缺陷)对聚(ADP-核糖)聚合酶-1(PARP-1)抑制剂的敏感性,PARP-1 是 HR 替代修复途径的关键酶。虽然有希望,但 PARP-1 抑制剂(PARPi)的治疗面临一些障碍,包括(1)获得性耐药,(2)寻找其他增敏的非 BRCA1/2 癌症缺陷,以及(3)缺乏预测 PARPi 反应的生物标志物。在这里,我们使用 PARP-1i 处理了来自乳腺癌、前列腺癌、结肠癌、胰腺癌和卵巢癌的 20 个人类细胞系来解决这些问题。Mre11-Rad50-Nbs1(MRN)复合物的异常使癌细胞对 PARP-1i 敏感,而 p53 状态的预测性较差,即使在与拓扑替康或电离辐射联合使用 PARP-1i 时也是如此。此外,分别监测 PARsylation 和 Rad51 焦点形成作为 PARP 活性和 HR 的替代标志物,支持它们作为 PARP-1i 反应生物标志物的候选物。至于耐药机制,我们证实了多药耐药外排转运蛋白的作用及其可逆性。更重要的是,我们证明了在人 BRCA1 突变型乳腺癌细胞中,shRNA 慢病毒介导的 53BP1 耗竭增加了它们对 PARP-1i 的耐药性。鉴于 BRCA 缺陷和三阴性乳腺癌中 53BP1 的优先丢失,我们的研究结果证明了在 PARPi 反应的候选预测生物标志物中评估 53BP1 的必要性。总体而言,这项研究有助于描述细胞对 PARP-1i 反应的遗传和功能决定因素,并有助于寻找生物标志物来利用 PARP 抑制剂进行癌症治疗。