TY - JOUR
T1 - The H3.3K27M oncohistone affects replication stress outcome and provokes genomic instability in pediatric glioma
AU - Bočkaj, Irena
AU - Martini, Tosca E.I.
AU - De Camargo Magalhães, Eduardo S.
AU - Bakker, Petra L.
AU - Meeuwsen-De Boer, Tiny G.J.
AU - Armandari, Inna
AU - Meuleman, Saskia L.
AU - Mondria, Marin T.
AU - Stok, Colin
AU - Kok, Yannick P.
AU - Bakker, Bjorn
AU - Wardenaar, René
AU - Seiler, Jonas
AU - Broekhuis, Mathilde J.C.
AU - Van Den Bos, Hilda
AU - Spierings, Diana C.J.
AU - Ringnalda, Femke C.A.
AU - Clevers, Hans
AU - Schüller, Ulrich
AU - Van Vugt, Marcel A.T.M.
AU - Foijer, Floris
AU - Bruggeman, Sophia W.M.
N1 - Publisher Copyright:
© 2021 Bočkaj et al.
PY - 2021/11/9
Y1 - 2021/11/9
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85119925929&partnerID=8YFLogxK
U2 - 10.1371/journal.pgen.1009868
DO - 10.1371/journal.pgen.1009868
M3 - Article
C2 - 34752469
AN - SCOPUS:85119925929
SN - 1553-7390
VL - 17
JO - PLoS Genetics
JF - PLoS Genetics
IS - 11
M1 - e1009868
ER -