Abstract P6-04-04: Maternal embryonic leucine zipper kinase (MELK) is a novel radiosensitizing and therapeutic target and is independently prognostic in triple-negative breast cancer

Abstract

Abstract Background: While effective targeted therapies exist for estrogen receptor (ER)-positive and HER2/neu-positive breast cancer, no such effective therapies exist for ER-negative, PR-negative, and HER2-negative (“triple negative”) cancers. Given the lack of targeted agents for triple negative (TN) disease and their relative radiation insensitivity, it is clear that additional targets for treatment are critically needed. Our previous work identified one such novel molecular target as maternal embryonic leucine zipper kinase (MELK), and we sought to investigate the impact of MELK expression on radiation response and patient outcomes. Methods: Using gene expression arrays, we interrogated the expression of MELK in 2,061 breast tumor samples as well as a panel of 51 breast cancer cell lines. We measured protein expression in TN cancers with western blotting and used clonogenic survival assays to quantitate radiosensitivity of BCC lines at baseline and after MELK inhibition. Multiple datasets were used to evaluate the prognostic import of MELK. Kaplan-Meier analysis using local control and survival data was performed. Chi squared scores were calculated to determine significance and hazard ratios (HR) and 95% confidence intervals (CI) were calculated. A Cox proportional hazards model was constructed to identify potential factors of survival. Results: We demonstrate that MELK expression is significantly elevated in human TN breast cancers, including chemoradiation resistant tumors (305 tumors compared to 1756 non-TN breast tumors; p-value 7.5 e-21). MELK protein and RNA expression is induced by ionizing radiation (5.6-7.5 fold at 72 hours, p-value <0.01). We characterized the radiation sensitivity of BCC lines and demonstrated that MELK expression is significantly correlated with radioresistance (as measured by clonogenic survival) in 21 breast cancer cell lines (R: 0.62, p-value 0.003). Inhibition of MELK using both siRNA and small molecule inhibitors induces radiation sensitivity in vitro with and enhancement ratio (ER) of 1.5-1.6. We demonstrate that high MELK expression is strongly correlated with p53 mutation positive status (p-value <0.001). Finally, local control and survival analyses of patients with BC showed that those patients whose tumors have high expression of MELK have significantly higher rates of LR after radiation and an overall poorer prognosis than patients with low expression of MELK (HR for LR 1.89-2.23, p-value 0.001; HR for overall survival 1.46-3.3; p-value <0.001 in 3 independent datasets). In multivariate analysis of all patients, only MELK expression and grade were significantly associated with worse local recurrence free (LRF) survival with a HR of 1.35 (95% CI 1.05-1.72, p-value < 0.01). Conclusion: Here, we identify MELK as a potential biomarker of radioresistance and target for radiosensitization in triple negative breast cancers. MELK overexpression was associated with local failure across multiple data sets. MVA identified MELK as the strongest factor associated with poor local control. Our results support the rationale for developing clinical strategies to inhibit MELK as a novel target in triple-negative breast cancer. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-04-04.