The H3.3K27M oncohistone affects replication stress outcome and provokes genomic instability in pediatric glioma

Irena Bočkaj, Tosca E I Martini, Eduardo S de Camargo Magalhães, Petra L Bakker, Tiny G J Meeuwsen-de Boer, Inna Armandari, Saskia L Meuleman, Marin T Mondria, Colin Stok, Yannick P Kok, Bjorn Bakker, René Wardenaar, Jonas Seiler, Mathilde J C Broekhuis, Hilda van den Bos, Diana C J Spierings, Femke C A Ringnalda, Hans Clevers, Ulrich Schüller, Marcel A T M van VugtFloris Foijer*, Sophia W M Bruggeman*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

While comprehensive molecular profiling of histone H3.3 mutant pediatric high-grade glioma has revealed extensive dysregulation of the chromatin landscape, the exact mechanisms driving tumor formation remain poorly understood. Since H3.3 mutant gliomas also exhibit high levels of copy number alterations, we set out to address if the H3.3K27M oncohistone leads to destabilization of the genome. Hereto, we established a cell culture model allowing inducible H3.3K27M expression and observed an increase in mitotic abnormalities. We also found enhanced interaction of DNA replication factors with H3.3K27M during mitosis, indicating replication defects. Further functional analyses revealed increased genomic instability upon replication stress, as represented by mitotic bulky and ultrafine DNA bridges. This co-occurred with suboptimal 53BP1 nuclear body formation after mitosis in vitro, and in human glioma. Finally, we observed a decrease in ultrafine DNA bridges following deletion of the K27M mutant H3F3A allele in primary high-grade glioma cells. Together, our data uncover a role for H3.3 in DNA replication under stress conditions that is altered by the K27M mutation, promoting genomic instability and potentially glioma development.

Author summary

The childhood brain cancer high-grade glioma is a devastating disease that almost invariably ends in the death of the patient. The discovery that mutations in histone H3 variants are predominant in this type of cancer created great interest into the mechanism of transformation of these so-called oncohistones. Yet whereas most research is directed at understanding the role of mutant H3 in deregulating gene expression, we studied if and how it contributes to genomic instability-a research avenue that remains mostly unexplored. We discovered that the H3.3K27M oncohistone increases sensitivity to replication stress during S-phase, ultimately leading to mitotic aberrancies. This can explain why childhood high-grade glioma is among the most genomically unstable cancers.

Original languageEnglish
Article number1009868
Number of pages28
JournalPLoS genetics
Volume17
Issue number11
Early online date9-Nov-2021
DOIs
Publication statusPublished - Nov-2021

Keywords

  • INTRINSIC PONTINE GLIOMAS
  • COMMON FRAGILE SITES
  • HISTONE H3.3
  • MCM PROTEINS
  • HIGH-GRADE
  • CHROMOSOME SEGREGATION
  • HUMAN-CELLS
  • DNA-DAMAGE
  • CHROMATIN
  • CHAPERONE

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