Blood Res.  2020 Mar;55(1):17-26. 10.5045/br.2020.55.1.17.

Characteristics of DNMT3A mutations in acute myeloid leukemia

Affiliations
  • 1Department of Laboratory Medicine, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul, Korea. yonggoo@catholic.ac.kr, microkim@catholic.ac.kr
  • 2Catholic Genetic Laboratory Center, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul, Korea.
  • 3Cancer Research Institute, Division of Hematology, Department of Internal Medicine, Catholic Blood and Marrow Transplantation Center, College of Medicine, The Catholic University of Korea, Seoul, Korea.
  • 4Department of Research and Development, Genetree Research, Seoul, Korea.

Abstract

BACKGROUND
DNMT3A mutations occur in approximately 20% of AML cases and are associated with changes in DNA methylation. CDKN2B plays an important role in the regulation of hematopoietic progenitor cells and DNMT3A mutation is associated with CDKN2B promoter methylation. We analyzed the characteristics of DNMT3A mutations including their clinical significance in AML and their influence on promoter methylation and CDKN2B expression.
METHODS
A total of 142 adults, recently diagnosed with de novo AML, were enrolled in the study. Mutations in DNMT3A, CEBPA, and NPM1 were analyzed by bidirectional Sanger sequencing. We evaluated CDKN2B promoter methylation and expression using pyrosequencing and RT-qPCR.
RESULTS
We identified DNMT3A mutations in 19.7% (N=28) of enrolled patients with AML, which increased to 29.5% when analysis was restricted to cytogenetically normal-AML. Mutations were located on exons from 8-23, and the majority, including R882, were found to be present on exon 23. We also identified a novel frameshift mutation, c.1590delC, in AML with biallelic mutation of CEBPA. There was no significant difference in CDKN2B promoter methylation according to the presence or type of DNMT3A mutations. CDKN2B expression inversely correlated with CDKN2B promoter methylation and was significantly higher in AML with R882H mutation in DNMT3A. We demonstrated that DNMT3A mutation was associated with poor AML outcomes, especially in cytogenetically normal-AML. The DNMT3A mutation remained as the independent unfavorable prognostic factor after multivariate analysis.
CONCLUSION
We characterized DNMT3A mutations in AML and revealed the association between the DNMT3A mutation and CDKN2B expression and clinical outcome.

Keyword

R882H; Methylation; CDKN2B; DNMT3A

MeSH Terms

Adult
DNA Methylation
Exons
Frameshift Mutation
Hematopoietic Stem Cells
Humans
Leukemia, Myeloid, Acute*
Methylation
Multivariate Analysis

Figure

  • Fig. 1 Location of 15 different exonic mutations of DNMT3A.

  • Fig. 2 Prognostic factors for patients with acute myeloid leukemia. (A) OS and (C) EFS in total patients with AML. (B) OS and (D) EFS in patients with CN-AML. (A) Kaplan-Meier survival curves for OS in 142 total patients with AML (P=0.0484) and (B) 61 patients with CN-AML (P=0.0376). (C) EFS curve in total 142 patients with AML (P=0.0012) and (D) EFS curve in 61 patients with CN-AML (P=0.0019).

  • Fig. 3 The results of quantitative analysis of DNMT3A R882H in eight patients with acute myeloid leukemia with regard to the WHO 2016 classification.


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