Therapy induced rewiring of signalling networks often lead to acquirement of platinum-resistance, thereby necessitating the use of non-platinum agents as second-line treatment particularly for epithelial ovarian cancer (EOC). A prior subject-specific assessment can guide the choice of optimal non-platinum agent/s and possible targeted therapeutic/s. Assessment of protein-protein interactions are superior to simple cytotoxicity assays to determine therapeutic efficacy and associated molecular responses. Utilizing improved PIP3-AKT and ERK1/2 activation Bioluminescence Resonance Energy Transfer (BRET) sensors, we report chemotherapy-induced ERK1/2 activation predominantly in cisplatin-paclitaxel resistant EOC cells and increased activation of both ERK1/2 and AKT in malignant ascites derived cancer cells from platinum-resistant patients but not from treatment-naive or platinum-sensitive relapse patients. Further, majority of the non-platinum drugs except irinotecan increased ERK1/2 activation in platinum-taxol resistant cells as observed by live-cell BRET assessment which were associated with p90RSK1/2 and BAD activation along with upregulation of multidrug transporter gene ABCC1 and cell survival genes like cyclin D1 and Bcl2. Interestingly, only irinotecan was able to sensitize these resistant cells. Altogether, this first report of BRET based sensing of molecular pathway activations in platinum resistant cell lines and patient's derived cancer cells highlight the clinical potential of BRET sensors in management of therapy resistant cancer.

治疗诱导的信号网络重新布线通常会导致获得铂耐药性，因此需要使用非铂药物作为二线治疗，特别是对于上皮性卵巢癌 (EOC)。先前的特定主题评估可以指导最佳非铂药物和可能的靶向治疗的选择。评估蛋白质-蛋白质相互作用优于简单的细胞毒性测定，以确定治疗效果和相关的分子反应。利用改进的 PIP3-AKT 和 ERK1/2 激活生物发光共振能量转移 (BRET) 传感器，我们报告了化疗诱导的 ERK1/2 激活主要在顺铂-紫杉醇耐药的 EOC 细胞中，并且在来自铂耐药患者的恶性腹水癌细胞中 ERK1/2 和 AKT 的激活增加，但不是来自未治疗或对铂敏感的复发患者. 此外，通过活细胞 BRET 评估观察到，除伊立替康以外的大多数非铂类药物增加了铂类紫杉醇耐药细胞中 ERK1/2 的激活，这与 p90RSK1/2 和 BAD 激活以及多药转运蛋白基因 ABCC1 和细胞的上调有关。细胞周期蛋白 D1 和 Bcl2 等存活基因。有趣的是，只有伊立替康能够使这些耐药细胞敏感。总之，这是第一份基于 BRET 检测铂耐药细胞系和患者分子通路激活的报告。