The novel, recurrent mutation in theTOP2Agene results in the enhanced topoisomerase activity and transcription deregulation in glioblastoma

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 inTP53,IDH1,ATRX,EGFRgenes found in this cohort, we identified a novel, recurrent mutation in theTOP2Agene 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 mutatedTOP2Ahad shorter overall survival than those with the wild typeTOP2A. Computational analyses of transcriptomic data showed that GBMs with the mutatedTOP2Ahave 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 discoveredTOP2Avariant is gain–of-function mutation.

Key points

• The most frequent genetic alterations in high grade gliomas are reported.

• A new mutation in theTOP2Agene was found in 4 patients from Polish population.

• A E948Q substitution changes TOP2A activities towards DNA.

• The recurrentTOP2Avariant 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 theTOP2Agene encoding topoisomerase 2A in glioblastomas. The TOP2A protein variant shows a higher affinity towards DNA and causes transcriptional alterations, suggesting a higherde novotranscription rate.