Please use this identifier to cite or link to this item: doi:10.22028/D291-36386
Title: Bisulfite profiling of the MGMT promoter and comparison with routine testing in glioblastoma diagnostics
Author(s): Tierling, Sascha
Jürgens-Wemheuer, Wiebke M.
Leismann, Alea
Becker-Kettern, Julia
Scherer, Michael
Wrede, Arne
Breuskin, David
Urbschat, Steffi
Sippl, Christoph
Oertel, Joachim
Schulz-Schaeffer, Walter J.
Walter, Jörn
Language: English
Title: Clinical Epigenetics
Volume: 14
Issue: 1
Publisher/Platform: BMC
Year of Publication: 2022
Free key words: DNA methylation
Local deep bisulfte sequencing
DDC notations: 500 Science
610 Medicine and health
Publikation type: Journal Article
Abstract: Background: Promoter methylation of the DNA repair gene O6 -methylguanine-DNA methyltransferase (MGMT) is an acknowledged predictive epigenetic marker in glioblastoma multiforme and anaplastic astrocytoma. Patients with methylated CpGs in the MGMT promoter beneft from treatment with alkylating agents, such as temozolomide, and show an improved overall survival and progression-free interval. A precise determination of MGMT promoter methyla‑ tion is of importance for diagnostic decisions. We experienced that diferent methods show partially divergent results in a daily routine. For an integrated neuropathological diagnosis of malignant gliomas, we therefore currently apply a combination of methylation-specifc PCR assays and pyrosequencing. Results: To better rationalize the variation across assays, we compared these standard techniques and assays to deep bisulfte sequencing results in a cohort of 80 malignant astrocytomas. Our deep analysis covers 49 CpG sites of the expanded MGMT promoter, including exon 1, parts of intron 1 and a region upstream of the transcription start site (TSS). We observed that deep sequencing data are in general in agreement with CpG-specifc pyrosequencing, while the most widely used MSP assays published by Esteller et al. (N Engl J Med 343(19):1350–1354, 2000. 10.1056/NEJM200011093431901) and Felsberg et al. (Clin Cancer Res 15(21):6683–6693, 2009. 1078-0432.CCR-08-2801) resulted in partially discordant results in 22 tumors (27.5%). Local deep bisulfte sequencing (LDBS) revealed that CpGs located in exon 1 are suited best to discriminate methylated from unmethylated samples. Based on LDBS data, we propose an optimized MSP primer pair with 83% and 85% concordance to pyrosequencing and LDBS data. A hitherto neglected region upstream of the TSS, with an overall higher methylation compared to exon 1 and intron 1 of MGMT, is also able to discriminate the methylation status. Conclusion: Our integrated analysis allows to evaluate and redefne co-methylation domains within the MGMT pro‑ moter and to rationalize the practical impact on assays used in daily routine diagnostics.
DOI of the first publication: 10.1186/s13148-022-01244-4
URL of the first publication:
Link to this record: urn:nbn:de:bsz:291--ds-363867
ISSN: 1868-7083
Date of registration: 8-Jun-2022
Description of the related object: Supplementary Information
Related object:
Faculty: M - Medizinische Fakultät
NT - Naturwissenschaftlich- Technische Fakultät
Department: M - Neurochirurgie
M - Neuropathologie
NT - Biowissenschaften
Professorship: M - Prof. Dr. Joachim Oertel
M - Prof. Dr. Walter Schulz-Schaeffer
NT - Prof. Dr. Jörn Walter
Collections:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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