A DNMT3A mutation common in AML exhibits dominant-negative effects in murine ES cells

Kim, Soo Jin; Zhao, Hongbo; Hardikar, Swanand; Singh, Anup K.; Goodell, Margaret A.; Chen, Taiping

doi:10.1182/blood-2013-02-483487

Cited by 150 publications

(134 citation statements)

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“…Heterodimerization of DNMT3A with DNMT3L enhances its methyltransferase activity [23,24]. Although mouse Dnmt3a R878H (corresponding to human R882H) mutant protein can still interact with wild-type Dnmt3a and Dnmt3b, co-expression of wild-type and mutant form in murine embryonic stem (ES) cells led to inhibition of the wild-type DNA methylation ability, consistent with dominant-negative effect of DNMT3A R882H mutations [25]. Moreover, DNMT3A R882H mutant was shown to inhibit wild-type de novo methylation activity by disrupting the formation of its functional tetramers [26], further confirming the dominant-negative role of this mutant.…”

Section: Dnmt3amentioning

confidence: 91%

Epigenetic dysregulation of hematopoietic stem cells and preleukemic state

Kunimoto

Nakajima

2017

Int J Hematol

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Section: Dnmt3amentioning

confidence: 91%

Epigenetic dysregulation of hematopoietic stem cells and preleukemic state

Kunimoto

Nakajima

2017

Int J Hematol

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“…[1][2][3][4] DNMT3A mutations in AML are almost exclusively heterozygous, and approximately 60% affect the arginine at amino acid position 882 (R882) in the methyltransferase domain. R882-mutant DNMT3A is a hypomorphic protein that also inhibits the remaining WT DNMT3A, dramatically reducing cellular DNA methyltransferase activity 5,6 including HOX genes, are significantly changed in DNMT3A-mutant AML. 4,8,12 In addition to the DNA methylation changes in Dnmt3a 2/2 HSCs, chromatin immunoprecipitation (ChIP)-sequencing (seq) and RNAseq data revealed evidence of perturbations of histone modifications.…”

Section: Introductionmentioning

confidence: 99%

DOT1L as a therapeutic target for the treatment of DNMT3A-mutant acute myeloid leukemia

Rau

Rodriguez²,

Luo

et al. 2016

Blood

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110

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• Data from Dnmt3a2/2 mice implicate Dot1l as a critical mediator of the malignant gene expression program of Dnmt3a-mediated leukemia.• Pharmacologic inhibition of DOT1L exerts potent antileukemic activity in DNMT3A-mutant human acute myeloid leukemia in vitro and in vivo.Mutations in DNA methyltransferase 3A (DNMT3A) are common in acute myeloid leukemia and portend a poor prognosis; thus, new therapeutic strategies are needed. The likely mechanism by which DNMT3A loss contributes to leukemogenesis is altered DNA methylation and the attendant gene expression changes; however, our current understanding is incomplete. We observed that murine hematopoietic stem cells (HSCs) in which Dnmt3a had been conditionally deleted markedly overexpress the histone 3 lysine 79 (H3K79) methyltransferase, Dot1l. We demonstrate that Dnmt3a 2/2 HSCs have increased H3K79 methylation relative to wild-type (WT) HSCs, with the greatest increases noted at DNA methylation canyons, which are regions highly enriched for genes dysregulated in leukemia and prone to DNA methylation loss with Dnmt3a deletion. These findings led us to explore DOT1L as a therapeutic target for the treatment of DNMT3A-mutant AML. We show that pharmacologic inhibition of DOT1L resulted in decreased expression of oncogenic canyon-associated genes and led to dose-and time-dependent inhibition of proliferation, induction of apoptosis, cell-cycle arrest, and terminal differentiation in DNMT3A-mutant cell lines in vitro. We show in vivo efficacy of the DOT1L inhibitor EPZ5676 in a nude rat xenograft model of DNMT3A-mutant AML. DOT1L inhibition was also effective against primary patient DNMT3A-mutant AML samples, reducing colony-forming capacity (CFC) and inducing terminal differentiation in vitro. These studies suggest that DOT1L may play a critical role in DNMT3A-mutant leukemia. With pharmacologic inhibitors of DOT1L already in clinical trials, DOT1L could be an immediately actionable therapeutic target for the treatment of this poor prognosis disease. (Blood. 2016;128(7):971-981)

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“…This kind of DNMT3A mutation confers reduced methyltransferase activity and promotes the possibility of dominant-negative consequences compared with the wild-type (WT) allele (2,9,10). Moreover, the DNMT3A mutation causes aberrant DNA hypomethylation and up-regulates a series of target genes involved in AML pathogenesis (11)(12)(13).…”

mentioning

confidence: 99%

Conditional knockin of Dnmt3a R878H initiates acute myeloid leukemia with mTOR pathway involvement

Dai

Wang

Huang

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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DNMT3A is frequently mutated in acute myeloid leukemia (AML). To explore the features of human AML with the hotspot DNMT3A R882H mutation, we generated Dnmt3a R878H conditional knockin mice, which developed AML with enlarged Lin − Sca1 + cKit + cell compartments. The transcriptome and DNA methylation profiling of bulk leukemic cells and the single-cell RNA sequencing of leukemic stem/progenitor cells revealed significant changes in gene expression and epigenetic regulatory patterns that cause differentiation arrest and growth advantage. Consistent with leukemic cell accumulation in G 2 /M phase, CDK1 was up-regulated due to mTOR activation associated with DNA hypomethylation. Overexpressed CDK1-mediated EZH2 phosphorylation resulted in an abnormal trimethylation of H3K27 profile. The mTOR inhibitor rapamycin elicited a significant therapeutic response in Dnmt3a R878H/WT mice.have been identified in a subset of acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), and acute lymphoblastic leukemia (ALL), with DNMT3A R882 being the hotspot (1-4). Clinical features of most AML cases with DNMT3A mutations include preferential involvement of a monocytic lineage (AML-M4 and -M5 subtypes), thrombocytosis, onset at a relatively old age, and poor prognosis (2, 5, 6). Careful genotypephenotype correlations suggest that patients manifest DNMT3A mutations in preleukemic hematopoietic stem cells (HSCs)/multipotent progenitors (MPPs), which exhibit a competitive advantage over normal HSCs. Because these mutations occur at a very early stage among genetic abnormalities, they are likely involved in the development of leukemia (7,8).Functionally, the DNMT3A R882 mutation might disrupt epigenetic regulation. This kind of DNMT3A mutation confers reduced methyltransferase activity and promotes the possibility of dominant-negative consequences compared with the wild-type (WT) allele (2, 9, 10). Moreover, the DNMT3A mutation causes aberrant DNA hypomethylation and up-regulates a series of target genes involved in AML pathogenesis (11-13).In vivo animal tests have shown that the Dnmt3a gene plays an essential role in hematopoiesis regulation. Dnmt3a −/− HSCs expand remarkably, but their differentiation is inhibited when Dnmt3a is conditionally inactivated in the murine hematopoietic system; this phenomenon is consistent with a preleukemic state (7). Moreover, all lethally irradiated mice transplanted with Dnmt3a-deleted HSCs died within 1 y and were diagnosed with a spectrum of malignancies similar to those observed in patients carrying DNMT3A mutations, including MDS, AML, primary myelofibrosis, and ALL, suggesting that Dnmt3a functions as a tumor suppressor (7). With a second hit of mutations in various genes such as N-RAS, C-KIT, or FLT3 in Dnmt3a −/− mice, Dnmt3a deletion induces leukemic transformation (14, 15). Although these results indicate a major role of Dnmt3a deletion in facilitating the development of leukemia, the in vivo roles of DNMT3A mutants in leukemogenesis still need to be addressed. In our previous work, b...

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A DNMT3A mutation common in AML exhibits dominant-negative effects in murine ES cells

Cited by 150 publications

References 22 publications

Epigenetic dysregulation of hematopoietic stem cells and preleukemic state

Epigenetic dysregulation of hematopoietic stem cells and preleukemic state

DOT1L as a therapeutic target for the treatment of DNMT3A-mutant acute myeloid leukemia

Conditional knockin of Dnmt3a R878H initiates acute myeloid leukemia with mTOR pathway involvement

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