MGMT Promoter-Methylation Analysis of Glioblastoma Using Illumina’s Infinium HumanMethylation450K Array: Subventricularzone Infiltration and MGMT Promoter Methylation Are Independent Prognostic Factors for Survival

Abstract BACKGROUND The CeTeG/NOA-09 trial recently showed a survival benefit for combination chemotherapy with CCNU/TMZ in glioblastoma patients with a methylated MGMT promoter as determined by quantitative Methylation-Specific PCR (qMSP). Identifying patient subgroups with a pronounced benefit from this novel treatment is crucial. Here, we report on the prognostic and predictive value of MGMT promoter methylation ratio determined by qMSP and investigate the concordance of pyrosequencing (PSQ) and qMSP for patients in this trial. METHODS qMSP and PSQ were used for MGMT promoter methylation analysis. The mITT population of the CeTeG/NOA-09 trial was used for multivariate analysis including the parameters MGMT promoter methylation ratio, RPA class and study center. RESULTS Patients of the mITT population of the CeTeG/NOA-09 trial (n=129) with MGMT promoter methylation ratio greater than 4 (qMSP) showed a superior overall survival compared to patients with borderline methylation ratio of 2–4 (p=0.0251). In the latter patients, treatment with CCNU/TMZ did not show a survival benefit (p=0.924). Multivariate analysis with treatment arm, RPA class and study center as covariates did not confirm a prognostic or predictive value of MGMT promoter methylation ratio (qMSP) for patients of the mITT population (n=129, HR=0.88; 95% CI: 0.72 – 1.08) or patients with a ratio greater than 4 (n=117, HR =0.86; 95% CI: 0.69 – 1.07). In a subset of 49 trial patients, qMSP and PSQ showed not only a high qualitative (45/49; 91.8%), but also a high quantitative concordance rate (Spearman correlation, r=0.83, p< 0.0001). CONCLUSION Glioblastoma patients with borderline MGMT promoter methylation (qMSP ratio 2–4) do not seem to benefit from combination treatment with CCNU/TMZ. Thus, we propose a qMSP cut-off of 4 as a novel decision tool for clinicians. qMSP and PSQ show a high concordance rate indicating that a decision for combination therapy can also be based on PSQ results.

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RESULTS OF THE INTERNATIONAL INTERLABORATORY COMPARISON OF MGMT PROMOTER METHYLATION ANALYSIS INVOLVING TWENTY-THREE ACADEMIC CENTERS IN GERMANY, AUSTRIA AND THE NETHERLANDS

Neuro-Oncology ◽

10.1093/neuonc/nou209.30 ◽

2014 ◽

Vol 16 (suppl 3) ◽

pp. iii49-iii50 ◽

Cited By ~ 3

Author(s):

G. Reifenberger ◽

B. Malzkorn ◽

T. Acker ◽

M. Bettstetter ◽

R. Buslei ◽

...

Keyword(s):

The Netherlands ◽

Promoter Methylation ◽

Interlaboratory Comparison ◽

Mgmt Promoter Methylation ◽

Methylation Analysis ◽

Mgmt Promoter

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Optimization of Quantitative MGMT Promoter Methylation Analysis Using Pyrosequencing and Combined Bisulfite Restriction Analysis

Journal of Molecular Diagnostics ◽

10.2353/jmoldx.2007.060167 ◽

2007 ◽

Vol 9 (3) ◽

pp. 368-381 ◽

Cited By ~ 140

Author(s):

Thomas Mikeska ◽

Christoph Bock ◽

Osman El-Maarri ◽

Anika Hübner ◽

Denise Ehrentraut ◽

...

Keyword(s):

Promoter Methylation ◽

Restriction Analysis ◽

Mgmt Promoter Methylation ◽

Methylation Analysis ◽

Mgmt Promoter

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Extent of Resection, MGMT Promoter Methylation Status and Tumor Location Independently Predict Progression-Free Survival in Adult Sporadic Pilocytic Astrocytoma

Cancers ◽

10.3390/cancers11081072 ◽

2019 ◽

Vol 11 (8) ◽

pp. 1072 ◽

Cited By ~ 2

Author(s):

Jungk ◽

Reinhardt ◽

Warta ◽

Capper ◽

Deimling ◽

...

Keyword(s):

Prognostic Factors ◽

Promoter Methylation ◽

Methylation Status ◽

Progression Free Survival ◽

Tumor Location ◽

Mgmt Promoter Methylation ◽

Free Survival ◽

Promoter Methylation Status ◽

Mgmt Promoter ◽

Mgmt Promoter Methylation Status

In adults, pilocytic astrocytomas (PA) account for less than 2% of gliomas, resulting in uncertainty regarding the clinical course and optimal treatment, particularly in cases where gross total resection (GTR) could not be achieved. Moreover, information on molecular markers and their prognostic impact is sparse. In order to improve risk stratification, we analyzed our institutional series of 58 patients aged 17 years and older with histology-proven intracranial PA World Health Organization grade I for clinical and molecular prognosticators. Anaplastic and NF1-associated tumors were excluded. O-6-methylguanine-DNA methyltransferase (MGMT) promoter methylation status was determined by pyrosequencing or 450k/850k DNA methylation array. A univariate log-rank test and multivariate StepAIC were applied to identify prognostic factors. The median age was 30 years (range 17–66). Tumors were located in the cerebral/cerebellar hemispheres, midline structures and cerebello-pontine angle in 53%, 38% and 9%. MGMT promoter methylation was present in eight patients (14%). GTR (39/58 patients) significantly reduced the likelihood of tumor recurrence (p = 0.0001). Tumor relapse occurred in 16 patients (28%) after a median progression-free survival (PFS) of 135 months (range 6–153 months); there was one tumor-related death. PFS at 5 and 10 years was 67% and 53%. In multivariate analysis, PFS was significantly prolonged in patients with GTR (HR 0.1; CI 0.03–0.37; p < 0.001), unmethylated MGMT promoter (HR 0.18; CI 0.05–0.64; p = 0.009) and midline tumors (HR 0.21; CI 0.06–0.78; p = 0.02). In conclusion, MGMT promoter methylation status and tumor location were identified as novel prognostic factors in adult PAs, pointing at distinct molecular subtypes and detecting patients in need of close observance and intensified treatment.

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MGMT promoter methylation analysis for allocating combined CCNU / TMZ chemotherapy: Lessons learned from the CeTeG / NOA ‐09 trial

International Journal of Cancer ◽

10.1002/ijc.33363 ◽

2020 ◽

Author(s):

Theophilos Tzaridis ◽

Niklas Schäfer ◽

Johannes Weller ◽

Joachim‐Peter Steinbach ◽

Uwe Schlegel ◽

...

Keyword(s):

Promoter Methylation ◽

Mgmt Promoter Methylation ◽

Lessons Learned ◽

Methylation Analysis ◽

Mgmt Promoter

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Prognostic Value of Three Different Methods of MGMT Promoter Methylation Analysis in a Prospective Trial on Newly Diagnosed Glioblastoma

PLoS ONE ◽

10.1371/journal.pone.0033449 ◽

2012 ◽

Vol 7 (3) ◽

pp. e33449 ◽

Cited By ~ 78

Author(s):

Arne Christians ◽

Christian Hartmann ◽

Axel Benner ◽

Jochen Meyer ◽

Andreas von Deimling ◽

...

Keyword(s):

Promoter Methylation ◽

Prognostic Value ◽

Prospective Trial ◽

Mgmt Promoter Methylation ◽

Newly Diagnosed ◽

Methylation Analysis ◽

Mgmt Promoter ◽

Newly Diagnosed Glioblastoma

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A preliminary comprehensive molecular-based nomogram for individualized estimation of survival in patients with newly diagnosed glioblastoma utilizing global microRNA expression data.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.2044 ◽

2019 ◽

Vol 37 (15_suppl) ◽

pp. 2044-2044

Author(s):

Denise Fabian ◽

Erica Hlavin Bell ◽

Joseph P. McElroy ◽

Tiantian Cui ◽

Jessica L. Fleming ◽

...

Keyword(s):

Overall Survival ◽

Prognostic Factors ◽

Clinical Data ◽

Mirna Expression ◽

Promoter Methylation ◽

Mgmt Promoter Methylation ◽

Expression Data ◽

Newly Diagnosed ◽

Mgmt Methylation ◽

Mgmt Promoter

2044 Background: Glioblastoma (GBM) is the most aggressive and common primary brain tumor. Nomograms are prediction models that help form individualized risk scores for cancer patients, which are valuable for treatment decision-making. The aim of this study is to create a refined nomogram by including novel molecular variables beyond MGMT promoter methylation. Methods: Clinical data and miRNA expression data were obtained from 226 newly diagnosed GBM patients. Clinical data included age at diagnosis, sex, Karnofsky performance status (KPS), extent of resection, O6-methylguanine-DNA methyltransferase ( MGMT) promoter methylation status, IDH mutation status and overall survival. Due to low representation of less than 13 cases each, IDH mutant glioblastomas and patients submitted to biopsy-only were excluded. Total RNA was isolated from formalin-fixed paraffin-embedded (FFPE) tissues; miRNA expression was subsequently measured using the NanoString human miRNA v3a assay. A Cox regression model was developed using glmnet R package with the elastic net penalty while adjusting for known prognostic factors. A dichotomized genomic score was created by finding the optimal cutpoint (maximum association with survival) of the linear combination of the selected. A nomogram was generated using known clinical prognostic factors, specifically age, sex, KPS, and MGMT status along with the dichotomized genomic score. Results: Four novel miRNAs were found to significantly correlate with overall survival and were used to create the dichotomized miRNA genomic score (GS). This score split the cohort into a poor performing group (GS_high) and a better performing group (GS_low) (p = 0.0031). A final nomogram was created using the Cox proportional hazards model (Figure 1). Factors that correlated with improved survival included younger age, KPS > 70, MGMT methylation and a low genomic score. Conclusions: This study is a proof of concept demonstrating that integration of molecular variables beyond MGMT methylation improve existing nomograms to provide individualized information about patient prognosis. Future directions include a more comprehensive analysis, including proteomic and methylation data, and subsequent validation in an external cohort. Finally, network analysis integrating molecular signatures of poor performers will help identify therapeutic targets.

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Long-term survival in glioblastoma: methyl guanine methyl transferase (MGMT) promoter methylation as independent favourable prognostic factor

Radiology and Oncology ◽

10.1515/raon-2015-0041 ◽

2016 ◽

Vol 50 (4) ◽

pp. 394-401 ◽

Cited By ~ 23

Author(s):

Uros Smrdel ◽

Mara Popovic ◽

Matjaz Zwitter ◽

Emanuela Bostjancic ◽

Andrej Zupan ◽

...

Keyword(s):

Prognostic Factors ◽

Promoter Methylation ◽

Mgmt Promoter Methylation ◽

Term Survival ◽

Long Term Survival ◽

Survivor Group ◽

Significant Difference ◽

Mgmt Promoter ◽

Term Survivor

Abstract Background In spite of significant improvement after multi-modality treatment, prognosis of most patients with glioblastoma remains poor. Standard clinical prognostic factors (age, gender, extent of surgery and performance status) do not clearly predict long-term survival. The aim of this case-control study was to evaluate immuno-histochemical and genetic characteristics of the tumour as additional prognostic factors in glioblastoma. Patients and methods Long-term survivor group were 40 patients with glioblastoma with survival longer than 30 months. Control group were 40 patients with shorter survival and matched to the long-term survivor group according to the clinical prognostic factors. All patients underwent multimodality treatment with surgery, postoperative conformal radiotherapy and temozolomide during and after radiotherapy. Biopsy samples were tested for the methylation of MGMT promoter (with methylation specific polymerase chain reaction), IDH1 (with immunohistochemistry), IDH2, CDKN2A and CDKN2B (with multiplex ligation-dependent probe amplification), and 1p and 19q mutations (with fluorescent in situ hybridization). Results Methylation of MGMT promoter was found in 95% and in 36% in the long-term survivor and control groups, respectively (p < 0.001). IDH1 R132H mutated patients had a non-significant lower risk of dying from glioblastoma (p = 0.437), in comparison to patients without this mutation. Other mutations were rare, with no significant difference between the two groups. Conclusions Molecular and genetic testing offers additional prognostic and predictive information for patients with glioblastoma. The most important finding of our analysis is that in the absence of MGMT promoter methylation, longterm survival is very rare. For patients without this mutation, alternative treatments should be explored.

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