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Significance of O6-methyl guanine methyltransferase promoter methylation in high grade glioma patients: optimal cutoff point, CpG locus, and genetic assay

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Abstract

Purpose

To evaluate O6-methyl guanine methyltransferase (MGMT) promoter methylation status in high grade glioma patients and to identify the best cutoff point as well as the most predictive CpG loci for patients survival.

Method

Consecutive high grade glioma patients treated with surgical gross total resection followed by concomitant radiochemotherapy and adjuvant chemotherapy were included in this retrospective observational study. Methylation status of MGMT promoter CpG island of resected tumor tissue were evaluated using next generation sequencing assay. The outcomes were grouped as CpG 70–78, CpG 79–83, CpG 84–87, CpG 70–87, and whole promoter. Quantitative analyses were dichotomized as methylated or unmethylated based on the cutoff points set to %10, and methylation was further graded as <%10 unmethylated, %10–30 low-methylated, and %30–100 high-methylated.

Results

Total of 95 patients with the mean age of 51.50 ± 12.36 years were included in the study. Overall survival (OS) and progression free survival (PFS) were 14.53 ± 1.92 (95% CI 10.77–18.30) and 10.90 ± 2.05 (95% CI 6.89–14.92) months, respectively. MGMT promoter was methylated in 38.2% of cases and high-methylated in 10.5% of cases. Methylation status of MGMT promoter was recognized as a very powerful predictor of OS and PFS. In particular, high-methylation of CpG 79–83 and CpG 84–87 islands at promoter region were strongly associated with better survival outcomes (p < 0.05).

Conclusion

Our outcomes support the prognostic value of MGMT promoter methylation in patients with high grade glioma. Sequencing of whole promoter CpG islands demonstrated that methylation of particular CpG sites might predict clinical outcomes more precisely.

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Funding

The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.

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Authors and Affiliations

  1. Department of Neurosurgery, Unye State Hospital, Ordu, Turkey

    Murat Buyuktepe

  2. Department of Medical Genetics, Ankara University School of Medicine, Ankara, Turkey

    Ibrahim Kaplan

  3. Departement of Neurosurgery, Ankara University School of Medicine, Ankara, Turkey

    Eyup Bayatli & Hasan Caglar Ugur

  4. INTERGEN Genetic Diagnosis and Research Center, Ankara, Turkey

    Haldun Dogan

Authors
  1. Murat Buyuktepe
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  2. Ibrahim Kaplan
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  3. Eyup Bayatli
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  4. Haldun Dogan
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  5. Hasan Caglar Ugur
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Contributions

All authors contributed to the study conception and design. M.B. performed material preparation and data collection. H.D. performed next generation sequencing, İ.K. and M.B. interpretated and grouped the methylation status. M.B. performed statistical analyses and wrote the first draft of the manuscript. All authors commented on previous versions of the manuscript, read and approved the final manuscript.

Corresponding author

Correspondence to Murat Buyuktepe.

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Competing interests

The authors declare no competing interests.

Conflict of interest

The authors have no relevant financial or non-financial interests to disclose.

Compliance with ethical standards

The study was approved by the Institutional Review Board of Ankara University and was conducted according to the tenets of the Declaration of Helsinki. (date: 13.03.2020; number: İ3-170-20 ) Written informed consent was obtained from each patient or patient’s guardian before enrollment.

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11060_2023_4397_MOESM1_ESM.jpg

Sup 1. MGMT gene (NM_002412) promoter/first exon region (hg19, chr10:131265097–131,265,807). green: promoter, purple: first exon (untranslated), blue: first intron, red: CpG’s, grey highlight: primer regions, blue highlight: non-CpG C’s

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Buyuktepe, M., Kaplan, I., Bayatli, E. et al. Significance of O6-methyl guanine methyltransferase promoter methylation in high grade glioma patients: optimal cutoff point, CpG locus, and genetic assay. J Neurooncol 164, 171–177 (2023). https://doi.org/10.1007/s11060-023-04397-1

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