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 Vugt; Floris Foijer; Sophia W M Bruggeman

doi:10.1371/journal.pgen.1009868

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

Hubrecht Institute

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

24 Citations (Scopus)

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.

Original language	English
Pages (from-to)	e1009868
Journal	PLoS Genetics
Volume	17
Issue number	11
DOIs	https://doi.org/10.1371/journal.pgen.1009868
Publication status	Published - Nov 2021

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Bočkaj, I., Martini, T. E. I., de Camargo Magalhães, E. S., Bakker, P. L., Meeuwsen-de Boer, T. G. J., Armandari, I., Meuleman, S. L., Mondria, M. T., Stok, C., Kok, Y. P., Bakker, B., Wardenaar, R., Seiler, J., Broekhuis, M. J. C., van den Bos, H., Spierings, D. C. J., Ringnalda, F. C. A., Clevers, H., Schüller, U., ... Bruggeman, S. W. M. (2021). The H3.3K27M oncohistone affects replication stress outcome and provokes genomic instability in pediatric glioma. PLoS Genetics, 17(11), e1009868. https://doi.org/10.1371/journal.pgen.1009868

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

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 = "Irena Bo{\v c}kaj and Martini, {Tosca E I} and {de Camargo Magalh{\~a}es}, {Eduardo S} and Bakker, {Petra L} and {Meeuwsen-de Boer}, {Tiny G J} and Inna Armandari and Meuleman, {Saskia L} and Mondria, {Marin T} and Colin Stok and Kok, {Yannick P} and Bjorn Bakker and Ren{\'e} Wardenaar and Jonas Seiler and Broekhuis, {Mathilde J C} and {van den Bos}, Hilda and Spierings, {Diana C J} and Ringnalda, {Femke C A} and Hans Clevers and Ulrich Sch{\"u}ller and {van Vugt}, {Marcel A T M} and Floris Foijer and Bruggeman, {Sophia W M}",

year = "2021",

month = nov,

doi = "10.1371/journal.pgen.1009868",

language = "English",

volume = "17",

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Bočkaj, I, Martini, TEI, de Camargo Magalhães, ES, Bakker, PL, Meeuwsen-de Boer, TGJ, Armandari, I, Meuleman, SL, Mondria, MT, Stok, C, Kok, YP, Bakker, B, Wardenaar, R, Seiler, J, Broekhuis, MJC, van den Bos, H, Spierings, DCJ, Ringnalda, FCA, Clevers, H, Schüller, U, van Vugt, MATM, Foijer, F & Bruggeman, SWM 2021, 'The H3.3K27M oncohistone affects replication stress outcome and provokes genomic instability in pediatric glioma', PLoS Genetics, vol. 17, no. 11, pp. e1009868. https://doi.org/10.1371/journal.pgen.1009868

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

PY - 2021/11

Y1 - 2021/11

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.

U2 - 10.1371/journal.pgen.1009868

DO - 10.1371/journal.pgen.1009868

M3 - Article

C2 - 34752469

SN - 1553-7404

VL - 17

SP - e1009868

JO - PLoS Genetics

JF - PLoS Genetics

IS - 11

ER -

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

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