Rocio Rosas-Alonso1,2, Julian Colmenarejo-Fernandez3,4, Olga Pernia3,4, Carlos Rodriguez-Antolín3,4, Isabel Esteban4,5, Ismael Ghanem6, Dario Sanchez-Cabrero4, Itsaso Losantos-Garcia7, Sara Palacios-Zambrano8, Gema Moreno-Bueno8,9,10, Javier de Castro4,6, Virginia Martinez-Marin6, Inmaculada Ibanez-de-Caceres11,12. 1. Epigenetics Laboratory. INGEMM, Paseo La Castellana 261. Edificio Bloque Quirúrgico Planta -2. University Hospital La Paz, 28046, Madrid, Spain. rocio.rosas@salud.madrid.org. 2. Experimental Therapies and Novel Biomarkers in Cancer. IdiPAZ, Madrid, Spain. rocio.rosas@salud.madrid.org. 3. Epigenetics Laboratory. INGEMM, Paseo La Castellana 261. Edificio Bloque Quirúrgico Planta -2. University Hospital La Paz, 28046, Madrid, Spain. 4. Experimental Therapies and Novel Biomarkers in Cancer. IdiPAZ, Madrid, Spain. 5. Pathology Department, La Paz University Hospital, Madrid, Spain. 6. Medical Oncology Department, La Paz University Hospital, Madrid, Spain. 7. Biostatistics Unit, La Paz University Hospital. IdiPAZ, Madrid, Spain. 8. MD Anderson Cancer Center, Madrid, Spain. 9. Biochemistry Department, UAM/ IIBm (CSIC-UAM), IdiPaz, Fundación MD Anderson Internacional, Madrid, Spain. 10. CIBERONC, Madrid, Spain. 11. Epigenetics Laboratory. INGEMM, Paseo La Castellana 261. Edificio Bloque Quirúrgico Planta -2. University Hospital La Paz, 28046, Madrid, Spain. inma.ibanezca@salud.madrid.org. 12. Experimental Therapies and Novel Biomarkers in Cancer. IdiPAZ, Madrid, Spain. inma.ibanezca@salud.madrid.org.
Abstract
BACKGROUND: The promoter hypermethylation of the methylguanine-DNA methyltransferase gene is a frequently used biomarker in daily clinical practice as it is associated with a favorable prognosis in glioblastoma patients treated with temozolamide. Due to the absence of adequately standardized techniques, international harmonization of the MGMT methylation biomarker is still an unmet clinical need for the diagnosis and treatment of glioblastoma patients. RESULTS: In this study we carried out a clinical validation of a quantitative assay for MGMT methylation detection by comparing a novel quantitative MSP using double-probe (dp_qMSP) with the conventional MSP in 100 FFPE glioblastoma samples. We performed both technologies and established the best cutoff for the identification of positive-methylated samples using the quantitative data obtained from dp_qMSP. Kaplan-Meier curves and ROC time dependent curves were employed for the comparison of both methodologies. CONCLUSIONS: We obtained similar results using both assays in the same cohort of patients, in terms of progression free survival and overall survival according to Kaplan-Meier curves. In addition, the results of ROC(t) curves showed that dp_qMSP increases the area under curve time-dependent in comparison with MSP for predicting progression free survival and overall survival over time. We concluded that dp_qMSP is an alternative methodology compatible with the results obtained with the conventional MSP. Our assay will improve the therapeutic management of glioblastoma patients, being a more sensitive and competitive alternative methodology that ensures the standardization of the MGMT-biomarker making it reliable and suitable for clinical use.
BACKGROUND: The promoter hypermethylation of the methylguanine-DNA methyltransferase gene is a frequently used biomarker in daily clinical practice as it is associated with a favorable prognosis in glioblastoma patients treated with temozolamide. Due to the absence of adequately standardized techniques, international harmonization of the MGMT methylation biomarker is still an unmet clinical need for the diagnosis and treatment of glioblastoma patients. RESULTS: In this study we carried out a clinical validation of a quantitative assay for MGMT methylation detection by comparing a novel quantitative MSP using double-probe (dp_qMSP) with the conventional MSP in 100 FFPE glioblastoma samples. We performed both technologies and established the best cutoff for the identification of positive-methylated samples using the quantitative data obtained from dp_qMSP. Kaplan-Meier curves and ROC time dependent curves were employed for the comparison of both methodologies. CONCLUSIONS: We obtained similar results using both assays in the same cohort of patients, in terms of progression free survival and overall survival according to Kaplan-Meier curves. In addition, the results of ROC(t) curves showed that dp_qMSP increases the area under curve time-dependent in comparison with MSP for predicting progression free survival and overall survival over time. We concluded that dp_qMSP is an alternative methodology compatible with the results obtained with the conventional MSP. Our assay will improve the therapeutic management of glioblastoma patients, being a more sensitive and competitive alternative methodology that ensures the standardization of the MGMT-biomarker making it reliable and suitable for clinical use.
Authors: Pierre Bady; Davide Sciuscio; Annie-Claire Diserens; Jocelyne Bloch; Martin J van den Bent; Christine Marosi; Pierre-Yves Dietrich; Michael Weller; Luigi Mariani; Frank L Heppner; David R Mcdonald; Denis Lacombe; Roger Stupp; Mauro Delorenzi; Monika E Hegi Journal: Acta Neuropathol Date: 2012-07-19 Impact factor: 17.088
Authors: C A Eads; K D Danenberg; K Kawakami; L B Saltz; C Blake; D Shibata; P V Danenberg; P W Laird Journal: Nucleic Acids Res Date: 2000-04-15 Impact factor: 16.971
Authors: 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 Journal: Ann Oncol Date: 2015-06-25 Impact factor: 32.976
Authors: Inmaculada Ibanez de Caceres; Essel Dulaimi; Amanda M Hoffman; Tahseen Al-Saleem; Robert G Uzzo; Paul Cairns Journal: Cancer Res Date: 2006-05-15 Impact factor: 12.701
Authors: Wolfgang Wick; Michael Platten; Christoph Meisner; Jörg Felsberg; Ghazaleh Tabatabai; Matthias Simon; Guido Nikkhah; Kirsten Papsdorf; Joachim P Steinbach; Michael Sabel; Stephanie E Combs; Jan Vesper; Christian Braun; Jürgen Meixensberger; Ralf Ketter; Regine Mayer-Steinacker; Guido Reifenberger; Michael Weller Journal: Lancet Oncol Date: 2012-05-10 Impact factor: 41.316
Authors: Javier Andrés Soto; Carlos Rodríguez-Antolín; Olga Vera; Olga Pernía; Isabel Esteban-Rodríguez; Maria Dolores Diestro; Javier Benitez; Fátima Sánchez-Cabo; Rafael Alvarez; Javier De Castro; Inmaculada Ibanez de Cáceres Journal: Clin Epigenetics Date: 2021-08-28 Impact factor: 6.551