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Impact on prognosis of the regional distribution of MGMT methylation with respect to the CpG island methylator phenotype and age in glioma patients

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Abstract

Clinical and molecular prognostic factors in gliomas include age, IDH mutation, the glioma CpG island methylator phenotype (G-CIMP+) and promoter methylation of the O6-methylguanine DNA-methyltransferase (MGMT) gene. Among these markers, a predictive value was reported in glioblastomas (GBM) for MGMT promoter methylation, in particular in elderly GBM patients. In this study, methylation data from 46 glioma samples with the Illumina 450K platform were obtained and extended using external data to include a total of 247 glioma samples. Methylation analysis of the whole MGMT gene with this platform revealed two strongly survival-associated CpG regions within the promoter and the gene body, which were confirmed in a reported dataset of high grade-gliomas. Methylation at the promoter (CpG 25, cg12981137 and the prognostic model MGMT-STP27) and at the gene body CpG 165 (cg07933035), were significantly associated with better overall survival, and strongly correlated with G-CIMP+ status. In this series, the prognostic value of MGMT methylation at the promoter was not observed in G-CIMP− cases, although around 50 % of them were MGMT-methylated. These results were also obtained in an homogeneously-treated series of chemoradiated G-CIMP− GBMs analyzed by MSP and qMSP, and confirmed in a reported pyrosequencing-analyzed series of gliomas. Interestingly, in contrast to the MGMT promoter, gene body methylation was of prognostic value in G-CIMP−patients older than 65 years. Our study highlights the relevance of the prognostic value of the different regions of methylation throughout the MGMT gene that could be affected by specific G-CIMP profiles and age groups.

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Acknowledgments

We acknowledge the Spanish Tumour Bank Network of the Centro Nacional de Investigaciones Oncológicas (Madrid, Spain) and the Tumor Bank of the Hospital Virgen de la Salud (BioB-HVS, Toledo, Spain) for providing tumor samples. This work was supported by Grants PI10/01974, PI10/01972, PI13/00800 and PI13/00055 from the Fondo de Investigaciones Sanitarias of the Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación, cofounded by the Fondo Europeo de Desarrollo Regional (FEDER 2007-2013).

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The authors have no conflicts of interest to declare.

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Authors and Affiliations

  1. Molecular Pathology Research Unit, Department of Pathology, Virgen de la Salud Hospital, Avda Barber 30, 45004, Toledo, Spain

    Pilar Mur, Teresa Hernández-Iglesias & Bárbara Meléndez

  2. Department of Neurosurgery, Virgen de la Salud Hospital, Avda Barber 30, 45004, Toledo, Spain

    Ángel Rodríguez de Lope

  3. Department of Pathology, Virgen de la Salud Hospital, Avda Barber 30, 45004, Toledo, Spain

    Francisco Javier Díaz-Crespo & Manuela Mollejo

  4. Department of Pathology, University Clinic Hospital, C/Villarroel 170, 08036, Barcelona, Spain

    Teresa Ribalta

  5. Department of Pathology, University Hospital Complex of Vigo (C.H.U.V.I), Rua Pizarro 22, 36204, Vigo, Spain

    Concepción Fiaño

  6. Department of Pathology, MD Anderson International, C/Arturo Soria 270, 28033, Madrid, Spain

    Juan Fernando García

  7. IdiPaz Research Unit, La Paz University Hospital, Pº de la Castellana 261, 28046, Madrid, Spain

    Juan Antonio Rey

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  1. Pilar Mur
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Supplementary Fig. 1

Principal component analysis (PCA) and hierarchical clustering using the CpGs with the highest variance (σ = 0.5, 1,268 probes). Two groups of tumors with different methylation profiles (orange G-CIMP+ tumors; green G-CIMP− tumors) are shown

Supplementary Fig. 2

Kaplan–Meier survival curves of glioma patients by G-CIMP status, grade, subtype, age, MGMT promoter (CpG 25) and gene body (CpG 165) methylation (P < 0.0001). ms median survival (days)

Supplementary Fig. 3

a Kaplan–Meier survival curves of the CpG 25 methylation status analyzed by pyrosequencing in 166 primary GBMs with wild-type IDH reported by Mulholland et al. (P > 0.05). b Kaplan–Meier survival curves of 68 GBM chemoradiated patients analyzed by means of MSP and qMSP, comparing MGMT methylated and unmethylated patients by age group (<60 or ≥60 years) (P > 0.05). Blue line MGMT methylated (M); green line MGMT unmethylated (UM); SE, standard error; ms median survival; dashed line ≥60 years; solid line <60 years

Supplementary Fig. 4

Kaplan–Meier survival curves of the CpG165 methylation status on G-CIMP− GBM patients by age group (<65 or ≥65 years). *P < 0.05. Blue line MGMT methylated (M); green line MGMT unmethylated (UM); Dashed line ≥65 years; solid line <65 years; ms median survival (days); SE standard error

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Mur, P., Rodríguez de Lope, Á., Díaz-Crespo, F.J. et al. Impact on prognosis of the regional distribution of MGMT methylation with respect to the CpG island methylator phenotype and age in glioma patients. J Neurooncol 122, 441–450 (2015). https://doi.org/10.1007/s11060-015-1738-9

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