Abstract
Background High grade gliomas (HGGs) are aggressive, primary brain tumors with poor clinical outcomes. We aim to better understand glioma pathobiology and find potential therapeutic susceptibilities.
Methods We designed a custom panel of 664 cancer- and epigenetics-related genes, and employed targeted next generation sequencing to study the genomic landscape of somatic and germline variants in 182 gliomas of different malignancy grades. mRNA sequencing was performed to detect transcriptomic abnormalities.
Results In addition to known alterations in TP53, IDH1, ATRX, EGFR genes found in this cohort, we identified a novel, recurrent mutation in the TOP2A gene coding for Topoisomerase 2A occurring only in glioblastomas (GBM, WHO grade IV gliomas). Biochemical assays with recombinant proteins demonstrated stronger DNA binding and DNA supercoil relaxation activities of the variant proteins. GBM patients carrying the mutated TOP2A had shorter overall survival than those with the wild type TOP2A. Computational analyses of transcriptomic data showed that GBMs with the mutated TOP2A have different transcriptomic patterns suggesting higher transcriptomic activity.
Conclusion We identified a novel TOP2A E948Q variant that strongly binds to DNA and is more active than the wild type protein. Our findings suggest that the discovered TOP2A variant is gain–of-function mutation.
Key points
The most frequent genetic alterations in high grade gliomas are reported.
A new mutation in the TOP2A gene was found in 4 patients from Polish population.
A E948Q substitution changes TOP2A activities towards DNA.
The recurrent TOP2A variant is a gain-of-function mutation.
Importance of the study Glioblastoma is a deadly disease. Despite recent advancements in genomics and innovative targeted therapies, glioblastoma therapy has not shown improvements. Insights into glioblastoma biology may improve diagnosis, prognosis, and treatment prediction, directing to a better outcome. We performed targeted sequencing of 664 cancer genes, and identified a new variant of the TOP2A gene encoding topoisomerase 2A in glioblastomas. The TOP2A protein variant shows a higher affinity towards DNA and causes transcriptional alterations, suggesting a higher de novo transcription rate.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
Data access: All processed sequencing data generated in this study have been submitted to The European Genome-phenome Archive (EGA; https://ega-archive.org/) under accession number EGAS00001004556.
Funding: Studies were supported by the Foundation for Polish Science TEAM-TECH Core Facility project “NGS platform for comprehensive diagnostics and personalized therapy in neuro-oncology”.
Conflict of interest: The authors declare that they have no conflict of interest.
Consent for publication: All co-authors have read and approved of its submission to this journal.
Authorship: Conception and design: B.W., B.Kam. Clinical data preparation: A.M, T.C., A.K., G.Z., A.S., M.K., C.S., R.C., M.Ba., W.Ka., W.S., M.Bu., B.C., M.Z., E.I.S., W. Kl., P.N. Experimental part: B.G., K.P., P.S., B.W., S.K.K., R.G., P.W, M.M, B.Kaz., J.C. Data interpretation: B.Kam., B.W., B.G., K.P, A.J.C. Data analysis: B.W., A..J.C. Manuscript writing: B.G., K.P., B.W., B.Kam. Final approval of manuscript: All authors. Accountable for all aspect of the study: All authors.