L Barault1, A Amatu2, F E Bleeker3, C Moutinho4, C Falcomatà1, V Fiano5, A Cassingena2, G Siravegna6, M Milione7, P Cassoni5, F De Braud8, R Rudà9, R Soffietti9, T Venesio1, A Bardelli10, P Wesseling11, P de Witt Hamer12, F Pietrantonio7, S Siena2, M Esteller13, A Sartore-Bianchi2, F Di Nicolantonio14. 1. Experimental Clinical Molecular Oncology, Candiolo Cancer Institute-FPO, IRCCS, Candiolo (Turin). 2. Niguarda Cancer Center, Ospedale Niguarda Ca' Granda, Milan, Italy. 3. Department of Clinical Genetics, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands. 4. Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), Barcelona, Spain. 5. Department of Medical Sciences, University of Turin, Città Della Salute e Della Scienza Hospital, Turin. 6. Experimental Clinical Molecular Oncology, Candiolo Cancer Institute-FPO, IRCCS, Candiolo (Turin); Department of Oncology, University of Torino, Candiolo (Turin); FIRC Institute of Molecular Oncology (IFOM), Milan. 7. Departments of Pathology and Laboratory Medicine. 8. Medical Oncology, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan. 9. Department of Neuro-Oncology, University of Turin and Città Della Salute e Della Scienza Hospital, Turin, Italy. 10. Experimental Clinical Molecular Oncology, Candiolo Cancer Institute-FPO, IRCCS, Candiolo (Turin); Department of Oncology, University of Torino, Candiolo (Turin). 11. Department of Pathology, VU University Medical Center, Amsterdam; Department of Pathology, Radboud University Medical Center, Nijmegen. 12. Department Neurosurgery, Neurosurgical Center Amsterdam, VU University Medical Center, Amsterdam, The Netherlands. 13. Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), Barcelona, Spain; Department of Physiological Sciences II, School of Medicine, University of Barcelona, Catalonia; Institucio Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain. 14. Experimental Clinical Molecular Oncology, Candiolo Cancer Institute-FPO, IRCCS, Candiolo (Turin); Department of Oncology, University of Torino, Candiolo (Turin). Electronic address: federica.dinicolantonio@unito.it.
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.
BACKGROUND:O(6)-methyl-guanine-methyl-transferase (MGMT) silencing by promoter methylation may identify cancerpatients 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 GBMpatients 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.
Authors: Patricia Mondelo-Macía; Ana María Rodríguez-Ces; María Mercedes Suárez-Cunqueiro; Laura Muinelo Romay Journal: Adv Exp Med Biol Date: 2022 Impact factor: 3.650
Authors: E Martín-Sánchez; E Pernaut-Leza; S Mendaza; A Cordoba; F Vicente-Garcia; I Monreal-Santesteban; J Pérez Vizcaino; M J Díaz De Cerio; N Perez-Janices; I Blanco-Luquin; D Escors; A Ulazia-Garmendia; D Guerrero-Setas Journal: Virchows Arch Date: 2016-04-21 Impact factor: 4.064
Authors: Andrea Sartore-Bianchi; Federica Di Nicolantonio; Ludovic Barault; Alessio Amatu; Giulia Siravegna; Agostino Ponzetti; Sebastian Moran; Andrea Cassingena; Benedetta Mussolin; Chiara Falcomatà; Alexandra M Binder; Carmen Cristiano; Daniele Oddo; Simonetta Guarrera; Carlotta Cancelliere; Sara Bustreo; Katia Bencardino; Sean Maden; Alice Vanzati; Patrizia Zavattari; Giuseppe Matullo; Mauro Truini; William M Grady; Patrizia Racca; Karin B Michels; Salvatore Siena; Manel Esteller; Alberto Bardelli Journal: Gut Date: 2017-10-05 Impact factor: 23.059