Digital PCR quantification of MGMT methylation refines prediction of clinical benefit from alkylating agents in glioblastoma and metastatic colorectal cancer
L Barault, A Amatu, F E Bleeker, C Moutinho, C Falcomatà, V Fiano, A Cassingena, G Siravegna, M Milione, P Cassoni, F De Braud, R Rudà, R Soffietti, T Venesio, A Bardelli, P Wesseling, P de Witt Hamer, F Pietrantonio, S Siena, M Esteller, A Sartore-Bianchi, F Di Nicolantonio, L Barault, A Amatu, F E Bleeker, C Moutinho, C Falcomatà, V Fiano, A Cassingena, G Siravegna, M Milione, P Cassoni, F De Braud, R Rudà, R Soffietti, T Venesio, A Bardelli, P Wesseling, P de Witt Hamer, F Pietrantonio, S Siena, M Esteller, A Sartore-Bianchi, F Di Nicolantonio
Abstract
Background: O(6)-methyl-guanine-methyl-transferase (MGMT) silencing by promoter methylation may identify cancer patients responding to the alkylating agents dacarbazine or temozolomide.
Patients and methods: We evaluated the prognostic and predictive value of MGMT methylation testing both in tumor and cell-free circulating DNA (cfDNA) from plasma samples using an ultra-sensitive two-step digital PCR technique (methyl-BEAMing). Results were compared with two established techniques, methylation-specific PCR (MSP) and Bs-pyrosequencing.
Results: Thresholds for MGMT methylated status for each technique were established in a training set of 98 glioblastoma (GBM) patients. The prognostic and the predictive value of MGMT methylated status was validated in a second cohort of 66 GBM patients treated with temozolomide in which methyl-BEAMing displayed a better specificity than the other techniques. Cutoff values of MGMT methylation specific for metastatic colorectal cancer (mCRC) tissue samples were established in a cohort of 60 patients treated with dacarbazine. In mCRC, both quantitative assays methyl-BEAMing and Bs-pyrosequencing outperformed MSP, providing better prediction of treatment response and improvement in progression-free survival (PFS) (P < 0.001). Ability of methyl-BEAMing to identify responding patients was validated in a cohort of 23 mCRC patients treated with temozolomide and preselected for MGMT methylated status according to MSP. In mCRC patients treated with dacarbazine, exploratory analysis of cfDNA by methyl-BEAMing showed that MGMT methylation was associated with better response and improved median PFS (P = 0.008).
Conclusions: Methyl-BEAMing showed high reproducibility, specificity and sensitivity and was applicable to formalin-fixed paraffin-embedded tissues and cfDNA. This study supports the quantitative assessment of MGMT methylation for clinical purposes since it could refine prediction of response to alkylating agents.
Keywords: DNA methylation; MGMT; alkylating agent; cell free circulating DNA; digital PCR; metastatic colorectal cancer.
© The Author 2015. Published by Oxford University Press on behalf of the European Society for Medical Oncology. All rights reserved. For permissions, please email: journals.permissions@oup.com.
Source: PubMed