Assessment of Quantitative and Allelic MGMT Methylation Patterns as a Prognostic Marker in Glioblastoma

Lasse S Kristensen; Signe R Michaelsen; Henrik Dyrbye; Derya Aslan; Kirsten Grunnet; Ib J Christensen; Hans S Poulsen; Kirsten Grønbæk; Helle Broholm

doi:10.1093/jnen/nlv024

Assessment of Quantitative and Allelic MGMT Methylation Patterns as a Prognostic Marker in Glioblastoma

Lasse S Kristensen, Signe R Michaelsen, Henrik Dyrbye, Derya Aslan, Kirsten Grunnet, Ib J Christensen, Hans S Poulsen, Kirsten Grønbæk, Helle Broholm

18 Citations (Scopus)

Abstract

Methylation of the O(6)-methylguanine-DNA methyltransferase (MGMT) gene is a predictive and prognostic marker in newly diagnosed glioblastoma patients treated with temozolomide but how MGMT methylation should be assessed to ensure optimal detection accuracy is debated. We developed a novel quantitative methylation-specific PCR (qMSP) MGMT assay capable of providing allelic methylation data and analyzed 151 glioblastomas from patients receiving standard of care treatment (Stupp protocol). The samples were also analyzed by immunohistochemistry (IHC), standard bisulfite pyrosequencing, and genotyped for the rs1690252 MGMT promoter single nucleotide polymorphism. Monoallelic methylation was observed more frequently than biallelic methylation, and some cases with monoallelic methylation expressed the MGMT protein whereas others did not. The presence of MGMT methylation was associated with better overall survival (p = 0.006; qMSP and p = 0.002; standard pyrosequencing), and the presence of the protein was associated with worse overall survival (p = 0.009). Combined analyses of qMSP and standard pyrosequencing or IHC identified additional patients who benefited from temozolomide treatment. Finally, low methylation levels were also associated with better overall survival (p = 0.061; qMSP and p = 0.02; standard pyrosequencing). These data support the use of both MGMT methylation and MGMT IHC but not allelic methylation data as prognostic markers in patients with temozolomide-treated glioblastoma.

Original language	English
Journal	Journal of Neuropathology and Experimental Neurology
Volume	75
Issue number	3
Pages (from-to)	246-55
Number of pages	10
ISSN	0022-3069
DOIs	https://doi.org/10.1093/jnen/nlv024
Publication status	Published - Mar 2016

Keywords

Adult
Aged
Antineoplastic Agents, Alkylating
Brain Neoplasms
Chi-Square Distribution
DNA Methylation
DNA Modification Methylases
DNA Repair Enzymes
Dacarbazine
Female
Genetic Association Studies
Genotype
Glioblastoma
Humans
Male
Middle Aged
Polymorphism, Single Nucleotide
Promoter Regions, Genetic
Retrospective Studies
Survival Analysis
Tumor Suppressor Proteins
Young Adult
Journal Article
Research Support, Non-U.S. Gov't

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10.1093/jnen/nlv024

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title = "Assessment of Quantitative and Allelic MGMT Methylation Patterns as a Prognostic Marker in Glioblastoma",

abstract = "Methylation of the O(6)-methylguanine-DNA methyltransferase (MGMT) gene is a predictive and prognostic marker in newly diagnosed glioblastoma patients treated with temozolomide but how MGMT methylation should be assessed to ensure optimal detection accuracy is debated. We developed a novel quantitative methylation-specific PCR (qMSP) MGMT assay capable of providing allelic methylation data and analyzed 151 glioblastomas from patients receiving standard of care treatment (Stupp protocol). The samples were also analyzed by immunohistochemistry (IHC), standard bisulfite pyrosequencing, and genotyped for the rs1690252 MGMT promoter single nucleotide polymorphism. Monoallelic methylation was observed more frequently than biallelic methylation, and some cases with monoallelic methylation expressed the MGMT protein whereas others did not. The presence of MGMT methylation was associated with better overall survival (p = 0.006; qMSP and p = 0.002; standard pyrosequencing), and the presence of the protein was associated with worse overall survival (p = 0.009). Combined analyses of qMSP and standard pyrosequencing or IHC identified additional patients who benefited from temozolomide treatment. Finally, low methylation levels were also associated with better overall survival (p = 0.061; qMSP and p = 0.02; standard pyrosequencing). These data support the use of both MGMT methylation and MGMT IHC but not allelic methylation data as prognostic markers in patients with temozolomide-treated glioblastoma.",

keywords = "Adult, Aged, Antineoplastic Agents, Alkylating, Brain Neoplasms, Chi-Square Distribution, DNA Methylation, DNA Modification Methylases, DNA Repair Enzymes, Dacarbazine, Female, Genetic Association Studies, Genotype, Glioblastoma, Humans, Male, Middle Aged, Polymorphism, Single Nucleotide, Promoter Regions, Genetic, Retrospective Studies, Survival Analysis, Tumor Suppressor Proteins, Young Adult, Journal Article, Research Support, Non-U.S. Gov't",

author = "Kristensen, {Lasse S} and Michaelsen, {Signe R} and Henrik Dyrbye and Derya Aslan and Kirsten Grunnet and Christensen, {Ib J} and Poulsen, {Hans S} and Kirsten Gr{\o}nb{\ae}k and Helle Broholm",

note = "{\textcopyright} 2016 American Association of Neuropathologists, Inc.",

year = "2016",

month = mar,

doi = "10.1093/jnen/nlv024",

language = "English",

volume = "75",

pages = "246--55",

journal = "Journal of Neuropathology and Experimental Neurology",

issn = "0022-3069",

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TY - JOUR

T1 - Assessment of Quantitative and Allelic MGMT Methylation Patterns as a Prognostic Marker in Glioblastoma

AU - Kristensen, Lasse S

AU - Michaelsen, Signe R

AU - Dyrbye, Henrik

AU - Aslan, Derya

AU - Grunnet, Kirsten

AU - Christensen, Ib J

AU - Poulsen, Hans S

AU - Grønbæk, Kirsten

AU - Broholm, Helle

PY - 2016/3

Y1 - 2016/3

N2 - Methylation of the O(6)-methylguanine-DNA methyltransferase (MGMT) gene is a predictive and prognostic marker in newly diagnosed glioblastoma patients treated with temozolomide but how MGMT methylation should be assessed to ensure optimal detection accuracy is debated. We developed a novel quantitative methylation-specific PCR (qMSP) MGMT assay capable of providing allelic methylation data and analyzed 151 glioblastomas from patients receiving standard of care treatment (Stupp protocol). The samples were also analyzed by immunohistochemistry (IHC), standard bisulfite pyrosequencing, and genotyped for the rs1690252 MGMT promoter single nucleotide polymorphism. Monoallelic methylation was observed more frequently than biallelic methylation, and some cases with monoallelic methylation expressed the MGMT protein whereas others did not. The presence of MGMT methylation was associated with better overall survival (p = 0.006; qMSP and p = 0.002; standard pyrosequencing), and the presence of the protein was associated with worse overall survival (p = 0.009). Combined analyses of qMSP and standard pyrosequencing or IHC identified additional patients who benefited from temozolomide treatment. Finally, low methylation levels were also associated with better overall survival (p = 0.061; qMSP and p = 0.02; standard pyrosequencing). These data support the use of both MGMT methylation and MGMT IHC but not allelic methylation data as prognostic markers in patients with temozolomide-treated glioblastoma.

AB - Methylation of the O(6)-methylguanine-DNA methyltransferase (MGMT) gene is a predictive and prognostic marker in newly diagnosed glioblastoma patients treated with temozolomide but how MGMT methylation should be assessed to ensure optimal detection accuracy is debated. We developed a novel quantitative methylation-specific PCR (qMSP) MGMT assay capable of providing allelic methylation data and analyzed 151 glioblastomas from patients receiving standard of care treatment (Stupp protocol). The samples were also analyzed by immunohistochemistry (IHC), standard bisulfite pyrosequencing, and genotyped for the rs1690252 MGMT promoter single nucleotide polymorphism. Monoallelic methylation was observed more frequently than biallelic methylation, and some cases with monoallelic methylation expressed the MGMT protein whereas others did not. The presence of MGMT methylation was associated with better overall survival (p = 0.006; qMSP and p = 0.002; standard pyrosequencing), and the presence of the protein was associated with worse overall survival (p = 0.009). Combined analyses of qMSP and standard pyrosequencing or IHC identified additional patients who benefited from temozolomide treatment. Finally, low methylation levels were also associated with better overall survival (p = 0.061; qMSP and p = 0.02; standard pyrosequencing). These data support the use of both MGMT methylation and MGMT IHC but not allelic methylation data as prognostic markers in patients with temozolomide-treated glioblastoma.

KW - Adult

KW - Aged

KW - Antineoplastic Agents, Alkylating

KW - Brain Neoplasms

KW - Chi-Square Distribution

KW - DNA Methylation

KW - DNA Modification Methylases

KW - DNA Repair Enzymes

KW - Dacarbazine

KW - Female

KW - Genetic Association Studies

KW - Genotype

KW - Glioblastoma

KW - Humans

KW - Male

KW - Middle Aged

KW - Polymorphism, Single Nucleotide

KW - Promoter Regions, Genetic

KW - Retrospective Studies

KW - Survival Analysis

KW - Tumor Suppressor Proteins

KW - Young Adult

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1093/jnen/nlv024

DO - 10.1093/jnen/nlv024

M3 - Journal article

C2 - 26883115

SN - 0022-3069

VL - 75

SP - 246

EP - 255

JO - Journal of Neuropathology and Experimental Neurology

JF - Journal of Neuropathology and Experimental Neurology

IS - 3

ER -

Assessment of Quantitative and Allelic MGMT Methylation Patterns as a Prognostic Marker in Glioblastoma

Abstract

Keywords

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