Clinical validation of a novel quantitative assay for the detection of MGMT methylation in glioblastoma patients

Clin Epigenetics. 2021 Mar 9;13(1):52. doi: 10.1186/s13148-021-01044-2.

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.

Keywords: Dp_qMSP; Glioblastoma; MGMT methylation; MSP.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Antineoplastic Agents, Alkylating / therapeutic use
  • Biomarkers, Tumor / genetics*
  • Brain Neoplasms / mortality
  • Brain Neoplasms / pathology*
  • Cohort Studies
  • CpG Islands
  • DNA Methylation
  • DNA Modification Methylases / genetics
  • DNA Repair Enzymes / genetics
  • Epigenomics
  • Female
  • Glioblastoma / diagnosis*
  • Glioblastoma / drug therapy
  • Glioblastoma / genetics*
  • Glioblastoma / surgery
  • Humans
  • Kaplan-Meier Estimate
  • Male
  • Middle Aged
  • Polymerase Chain Reaction / methods
  • Polymerase Chain Reaction / trends
  • Prognosis
  • Progression-Free Survival
  • Promoter Regions, Genetic / genetics
  • Prospective Studies
  • ROC Curve
  • Sensitivity and Specificity
  • Temozolomide / therapeutic use
  • Tumor Suppressor Proteins / genetics

Substances

  • Antineoplastic Agents, Alkylating
  • Biomarkers, Tumor
  • Tumor Suppressor Proteins
  • DNA Modification Methylases
  • MGMT protein, human
  • DNA Repair Enzymes
  • Temozolomide