A Comparative Study of Methyl-BEAMing and Droplet Digital PCR for MGMT Gene Promoter Hypermethylation Detection

<b><i>Background:</i></b> <i>O-6-methylguanine-DNA methyltransferase</i> is responsible for the direct repair of O6-methylguanine lesions induced by alkylating agents, including temozolomide. <i>O-6-methylguanine-DNA methyltransferase</i> promoter hypermethylation is a well-established biomarker for temozolomide response in glioblastoma patients, also correlated with therapeutic response in colorectal cancer. <b><i>Objectives:</i></b> The ARETHUSA clinical trial aims to stratify colorectal cancer patients based on their mismatch repair status. Mismatch repair-deficient patients are eligible for treatment with immune checkpoint inhibitors (anti-PDL-1), whereas mismatch repair-proficient samples are screened for <i>O-6-methylguanine-DNA methyltransferase</i> promoter methylation to identify those suitable for temozolomide treatment. <b><i>Methods:</i></b> In this context, a subset of ARETHUSA metastatic colorectal cancer samples was used to compare two different techniques for assessing <i>O-6-methylguanine-DNA methyltransferase</i> hypermethylation: Methyl-BEAMing, a highly sensitive digital PCR approach that combines emulsion PCR and flow cytometry, and droplet digital PCR, a more automated procedure that enables the rapid, operator-independent analysis of a large number of samples. <b><i>Results:</i></b> Our study clearly demonstrates that the results obtained using Methyl-BEAMing and droplet digital PCR are comparable, with both techniques showing similar accuracy, sensitivity, and reproducibility. <b><i>Conclusions:</i></b> Digital droplet PCR proved to be an efficient method for detecting gene promoter methylation. However, the Methyl-BEAMing method has proved more sensitive for detecting low quantities of DNA.

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Journal
Diagnostics
Published
2024-11-05
DOI
https://doi.org/10.3390/diagnostics14222467
Citations
2
Primary Topic
Cancer Genomics and Diagnostics
Type
article
Field-Weighted Citation Impact
0.57
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article

A Comparative Study of Methyl-BEAMing and Droplet Digital PCR for MGMT Gene Promoter Hypermethylation Detection

Eleonora Gusmaroli, Sara E. Bellomo, Federica Di Nicolantonio, Emanuele Valtorta et al.
2 citations
Diagnostics
Cancer Genomics and Diagnostics
0.57
article

A Comparative Study of Methyl-BEAMing and Droplet Digital PCR for MGMT Gene Promoter Hypermethylation Detection

Eleonora Gusmaroli, Sara E. Bellomo, Federica Di Nicolantonio, Emanuele Valtorta, Alessandro Cavaliere, Alessandra Raimondi, Marco Macagno, Davide Ciardiello, Erica Bonazzina, Andrea Sartore‐Bianchi, Salvatore Siena, Ludovic Barault, Alessio Amatu, Federica Maione, Noemi Congiusta, Guido Serini, Nicola Personeni, Maria Costanza Aquilano, Lorenzo Gervaso, Katia Bencardino, Fariha Idrees, Filippo Pietrantonio, Valeria Pessei, Silvia Marsoni, Luca Lazzari, Armando Santoro, Emanuela Bonoldi, M.G. Zampino, Giuseppe Mondello
article en
2 citations

Abstract

<b><i>Background:</i></b> <i>O-6-methylguanine-DNA methyltransferase</i> is responsible for the direct repair of O6-methylguanine lesions induced by alkylating agents, including temozolomide. <i>O-6-methylguanine-DNA methyltransferase</i> promoter hypermethylation is a well-established biomarker for temozolomide response in glioblastoma patients, also correlated with therapeutic response in colorectal cancer. <b><i>Objectives:</i></b> The ARETHUSA clinical trial aims to stratify colorectal cancer patients based on their mismatch repair status. Mismatch repair-deficient patients are eligible for treatment with immune checkpoint inhibitors (anti-PDL-1), whereas mismatch repair-proficient samples are screened for <i>O-6-methylguanine-DNA methyltransferase</i> promoter methylation to identify those suitable for temozolomide treatment. <b><i>Methods:</i></b> In this context, a subset of ARETHUSA metastatic colorectal cancer samples was used to compare two different techniques for assessing <i>O-6-methylguanine-DNA methyltransferase</i> hypermethylation: Methyl-BEAMing, a highly sensitive digital PCR approach that combines emulsion PCR and flow cytometry, and droplet digital PCR, a more automated procedure that enables the rapid, operator-independent analysis of a large number of samples. <b><i>Results:</i></b> Our study clearly demonstrates that the results obtained using Methyl-BEAMing and droplet digital PCR are comparable, with both techniques showing similar accuracy, sensitivity, and reproducibility. <b><i>Conclusions:</i></b> Digital droplet PCR proved to be an efficient method for detecting gene promoter methylation. However, the Methyl-BEAMing method has proved more sensitive for detecting low quantities of DNA.

DiagnosticsVol. 14(22)
University of Milan (IT), Azienda Socio Sanitaria Territoriale Grande Ospedale Metropolitano Niguarda (IT), Casa di Cura Villa Garda (IT), IFOM (IT), Candiolo Cancer Institute (IT), Istituti di Ricovero e Cura a Carattere Scientifico (IT), Fondazione IRCCS Istituto Nazionale dei Tumori (IT), IRCCS Humanitas Research Hospital (IT), University of Turin (IT), European Institute of Oncology (IT)
Openalex Percentile: Top 30%
Cancer Genomics and Diagnostics
0.57
DOIOpen Access PDFOpenAlex
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