DNMT3A Harboring Leukemia-Associated Mutations Directs Sensitivity to DNA Damage at Replication Forks.
Journal
Clinical cancer research : an official journal of the American Association for Cancer Research
ISSN: 1557-3265
Titre abrégé: Clin Cancer Res
Pays: United States
ID NLM: 9502500
Informations de publication
Date de publication:
15 02 2022
15 02 2022
Historique:
received:
06
08
2021
revised:
10
10
2021
accepted:
27
10
2021
pubmed:
31
10
2021
medline:
8
4
2022
entrez:
30
10
2021
Statut:
ppublish
Résumé
In acute myeloid leukemia (AML), recurrent DNA methyltransferase 3A (DNMT3A) mutations are associated with chemoresistance and poor prognosis, especially in advanced-age patients. Gene-expression studies in DNMT3A-mutated cells identified signatures implicated in deregulated DNA damage response and replication fork integrity, suggesting sensitivity to replication stress. Here, we tested whether pharmacologically induced replication fork stalling, such as with cytarabine, creates a therapeutic vulnerability in cells with DNMT3A(R882) mutations. Leukemia cell lines, genetic mouse models, and isogenic cells with and without DNMT3A(mut) were used to evaluate sensitivity to nucleoside analogues such as cytarabine in vitro and in vivo, followed by analysis of DNA damage and signaling, replication restart, and cell-cycle progression on treatment and after drug removal. Transcriptome profiling identified pathways deregulated by DNMT3A(mut) expression. We found increased sensitivity to pharmacologically induced replication stress in cells expressing DNMT3A(R882)-mutant, with persistent intra-S-phase checkpoint activation, impaired PARP1 recruitment, and elevated DNA damage, which was incompletely resolved after drug removal and carried through mitosis. Pulse-chase double-labeling experiments with EdU and BrdU after cytarabine washout demonstrated a higher rate of fork collapse in DNMT3A(mut)-expressing cells. RNA-seq studies supported deregulated cell-cycle progression and p53 activation, along with splicing, ribosome biogenesis, and metabolism. Together, our studies show that DNMT3A mutations underlie a defect in recovery from replication fork arrest with subsequent accumulation of unresolved DNA damage, which may have therapeutic tractability. These results demonstrate that, in addition to its role in epigenetic control, DNMT3A contributes to preserving genome integrity during replication stress. See related commentary by Viny, p. 573.
Identifiants
pubmed: 34716195
pii: 1078-0432.CCR-21-2863
doi: 10.1158/1078-0432.CCR-21-2863
pmc: PMC8866212
mid: NIHMS1754276
doi:
Substances chimiques
DNMT3A protein, human
0
Dnmt3a protein, mouse
0
DNA Methyltransferase 3A
EC 2.1.1.37
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
756-769Subventions
Organisme : Center for Biotechnology Research
ID : RRID: SCR_019119
Organisme : NCI NIH HHS
ID : R00 CA178191
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK121831
Pays : United States
Organisme : NIH HHS
ID : CA188561
Pays : United States
Organisme : NCI NIH HHS
ID : R21 CA252400
Pays : United States
Organisme : NIDDK NIH HHS
ID : T32 DK074367
Pays : United States
Organisme : NCI NIH HHS
ID : K99 CA178191
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA188561
Pays : United States
Commentaires et corrections
Type : CommentIn
Informations de copyright
©2021 The Authors; Published by the American Association for Cancer Research.
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