Characterization of CEBPA mutations and promoter hypermethylation in pediatric acute myeloid leukemia

Iris H. I. M. Hollink, Marry M. van den Heuvel-Eibrink, Susan T. C. J. M. Arentsen-Peters, Martin Zimmermann, Justine K. Peeters, Peter J. M. Valk, Brian V. Balgobind, Edwin Sonneveld, Gertjan J. L. Kaspers, Eveline S. J. M. de Bont, Jan Trka, Andre Baruchel, Ursula Creutzig, Rob Pieters, Dirk Reinhardt, C. Michel Zwaan*

*Corresponding author for this work

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Abstract

Background

Dysfunctioning of CCAAT/enhancer binding protein alpha (C/EBP alpha) in acute myeloid leukemia can be caused, amongst others, by mutations in the encoding gene (CEBPA) and by promoter hypermethylation. CEBPA-mutated acute myeloid leukemia is associated with a favorable outcome, but this may be restricted to the case of double mutations in CEBPA in adult acute myeloid leukemia. In pediatric acute myeloid leukemia, data on the impact of these mutations are limited to one series, and data on promoter hypermethylation are lacking. Our objective was to investigate the characteristics, gene expression profiles and prognostic impact of the different CEBPA aberrations in pediatric acute myeloid leukemia.

Design and Methods

We screened a large pediatric cohort (n=252) for CEBPA single and double mutations by direct sequencing, and for promoter hypermethylation by methylation-specific polymerase chain reaction. Furthermore, we determined the gene-expression profiles (Affymetrix HGU133 plus 2.0 arrays) of this cohort (n=237).

Results

Thirty-four mutations were identified in 20 out of the 252 cases (7.9%), including 14 double-mutant and 6 single-mutant cases. CEBPA double mutations conferred a significantly better 5-year overall survival compared with single mutations (79% versus 25%, respectively; P=0.04), and compared with CEBPA wild-type acute myeloid leukemia excluding core-binding factor cases (47%; P=0.07). Multivariate analysis confirmed that the double mutations were an independent favorable prognostic factor for survival (hazard ratio 0.23, P=0.04). The combination of screening for promoter hypermethylation and gene expression profiling identified five patients with silenced CEBPA, of whom four cases relapsed. All cases characteristically expressed T-lymphoid markers. Moreover, unsupervised clustering of gene expression profiles showed a clustering of CEBPA double-mutant and silenced cases, pointing towards a common hallmark of abrogated C/EBP alpha-functioning in these acute myeloid leukemias.

Conclusions

We showed the independent favorable outcome of patients with CEBPA double-mutant acute myeloid leukemia in a large pediatric series. This molecular marker may, therefore, improve risk-group stratification in pediatric acute myeloid leukemia. For the first time, CEBPA-silenced cases are suggested to confer a poor outcome in pediatric acute myeloid leukemia, indicating that further investigation of this aberration is needed. Furthermore, clustering of gene expression profiles provided insight into the biological similarities and diversities of the different aberrations in CEBPA in pediatric acute myeloid leukemia.

Original languageEnglish
Pages (from-to)384-392
Number of pages9
JournalHaematologica
Volume96
Issue number3
DOIs
Publication statusPublished - Mar-2011

Keywords

  • Pediatric acute myeloid leukemia
  • CEBPA mutation
  • promoter hypermethylation
  • molecular marker
  • prognostic significance
  • BINDING-PROTEIN-ALPHA
  • ACUTE LYMPHOBLASTIC-LEUKEMIA
  • GENE-EXPRESSION PROFILES
  • CELLULAR-DRUG RESISTANCE
  • FACTOR-C/EBP-ALPHA
  • TRANSCRIPTION FACTOR
  • PROGNOSTIC-SIGNIFICANCE
  • INITIATING CELLS
  • AML
  • DIFFERENTIATION

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