DNMT3A Harboring Leukemia-Associated Mutations Directs Sensitivity to DNA Damage at Replication Forks.

Autor:	Venugopal K; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Feng Y; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Nowialis P; Department of Anatomy and Cell Biology, University of Florida College of Medicine, Gainesville, Florida., Xu H; Department of Pediatrics, Division of Infectious Diseases, University of Florida College of Medicine, Gainesville, Florida., Shabashvili DE; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Berntsen CM; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Kaur P; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Krajcik KI; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Taragjini C; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Zaroogian Z; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Casellas Román HL; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Posada LM; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Gunaratne C; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Li J; Department of Medicine, Division of Hematology/ Oncology, University of Florida College of Medicine, Gainesville, Florida., Dupéré-Richer D; Department of Medicine, Division of Hematology/ Oncology, University of Florida College of Medicine, Gainesville, Florida., Bennett RL; Department of Medicine, Division of Hematology/ Oncology, University of Florida College of Medicine, Gainesville, Florida.; University of Florida Health Cancer Center, Gainesville, Florida., Pondugula S; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Riva A; University of Florida Health Cancer Center, Gainesville, Florida.; Bioinformatics Core, Interdisciplinary Center for Biotechnology Research, University of Florida, Gainesville, Florida., Cogle CR; Department of Medicine, Division of Hematology/ Oncology, University of Florida College of Medicine, Gainesville, Florida.; University of Florida Health Cancer Center, Gainesville, Florida., Opavsky R; Department of Anatomy and Cell Biology, University of Florida College of Medicine, Gainesville, Florida.; University of Florida Health Cancer Center, Gainesville, Florida., Law BK; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida.; University of Florida Health Cancer Center, Gainesville, Florida., Bhaduri-McIntosh S; Department of Pediatrics, Division of Infectious Diseases, University of Florida College of Medicine, Gainesville, Florida.; University of Florida Health Cancer Center, Gainesville, Florida.; Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville, Florida., Kubicek S; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria., Staber PB; Division of Hematology and Hemostaseology, Department of Medicine 1, Comprehensive Cancer Center Vienna, Medical University of Vienna, Vienna, Austria., Licht JD; Department of Medicine, Division of Hematology/ Oncology, University of Florida College of Medicine, Gainesville, Florida.; University of Florida Health Cancer Center, Gainesville, Florida., Bird JE; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida., Guryanova OA; Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville, Florida.; University of Florida Health Cancer Center, Gainesville, Florida.
Jazyk:	angličtina
Zdroj:	Clinical cancer research : an official journal of the American Association for Cancer Research [Clin Cancer Res] 2022 Feb 15; Vol. 28 (4), pp. 756-769.
DOI:	10.1158/1078-0432.CCR-21-2863
Abstrakt:	Purpose: 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. Experimental Design: 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. Results: 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. Conclusions: 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. (©2021 The Authors; Published by the American Association for Cancer Research.)
Databáze:	MEDLINE
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