Abstract
Hypermethylation of the distal CEBPA promoter region has been reported to result in the downregulation of CEBPA expression in several malignancies. However, the clinical implication of CEBPA hypermethylation in acute myeloid leukemia (AML) remains unclear. To investigate the correlation between CEBPA hypermethylation and clinical features in AML, quantitative MassARRAY analyses for CEBPA methylation status were performed on a cohort of 193 patients. High CEBPA methylation group (CEBPAhigh-meth, n=28) and low methylation group (CEBPAlow-meth, n=165) were defined by using two-way hierarchical clustering. With a median follow-up of 48 months, among the 125 patients receiving standard induction therapy, CEBPAhigh-meth was associated with better treatment response (complete remission rate 93.3% versus 73.6%, P=0.116). In patients with normal karyotype and without CEBPA and NPM1 mutations, the CEBPAhigh-meth had longer overall survival (OS) than the CEBPAlow-meth (P=0.028). Multivariate analysis further supported that the CEBPA methylation was an independent prognostic factor for disease free-survival (hazard ratio=0.416; 95% confidence interval, 0.223–0.777, P=0.006) and OS (hazard ratio=0.406; 95% confidence interval, 0.166–0.996, P=0.050). We conclude that CEBPA methylation status is a useful prognostic biomarker for AML patients.
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References
Tenen DG . Disruption of differentiation in human cancer: AML shows the way. Nat Rev Cancer 2003; 3: 89–101.
Kelly LM, Gilliland DG . Genetics of myeloid leukemias. Annu Rev Genomics Hum Genet 2002; 3: 179–198.
Cammenga J, Mulloy JC, Berguido FJ, MacGrogan D, Viale A, Nimer SD . Induction of C/EBPalpha activity alters gene expression and differentiation of human CD34+ cells. Blood 2003; 101: 2206–2214.
Keeshan K, Santilli G, Corradini F, Perrotti D, Calabretta B . Transcription activation function of C/EBPalpha is required for induction of granulocytic differentiation. Blood 2003; 102: 1267–1275.
Tada Y, Brena RM, Hackanson B, Morrison C, Otterson GA, Plass C . Epigenetic modulation of tumor suppressor CCAAT/enhancer binding protein alpha activity in lung cancer. J Natl Cancer Inst 2006; 98: 396–406.
Bennett KL, Hackanson B, Smith LT, Morrison CD, Lang JC, Schuller DE et al. Tumor suppressor activity of CCAAT/enhancer binding protein alpha is epigenetically down-regulated in head and neck squamous cell carcinoma. Cancer Res 2007; 67: 4657–4664.
Hackanson B, Bennett KL, Brena RM, Jiang J, Claus R, Chen SS et al. Epigenetic modification of CCAAT/enhancer binding protein alpha expression in acute myeloid leukemia. Cancer Res 2008; 68: 3142–3151.
Pabst T, Mueller BU, Zhang P, Radomska HS, Narravula S, Schnittger S et al. Dominant-negative mutations of CEBPA, encoding CCAAT/enhancer binding protein-alpha (C/EBPalpha), in acute myeloid leukemia. Nat Genet 2001; 27: 263–270.
Lin LI, Chen CY, Lin DT, Tsay W, Tang JL, Yeh YC et al. Characterization of CEBPA mutations in acute myeloid leukemia: most patients with CEBPA mutations have biallelic mutations and show a distinct immunophenotype of the leukemic cells. Clin Cancer Res 2005; 11: 1372–1379.
Preudhomme C, Sagot C, Boissel N, Cayuela JM, Tigaud I, de Botton S et al. Favorable prognostic significance of CEBPA mutations in patients with de novo acute myeloid leukemia: a study from the Acute Leukemia French Association (ALFA). Blood 2002; 100: 2717–2723.
Barjesteh van Waalwijk van Doorn-Khosrovani S, Erpelinck C, Meijer J, van Oosterhoud S, van Putten WL, Valk PJ et al. Biallelic mutations in the CEBPA gene and low CEBPA expression levels as prognostic markers in intermediate-risk AML. Hematol J 2003; 4: 31–40.
Frohling S, Schlenk RF, Stolze I, Bihlmayr J, Benner A, Kreitmeier S et al. CEBPA mutations in younger adults with acute myeloid leukemia and normal cytogenetics: prognostic relevance and analysis of cooperating mutations. J Clin Oncol 2004; 22: 624–633.
Schlenk RF, Dohner K, Krauter J, Frohling S, Corbacioglu A, Bullinger L et al. Mutations and treatment outcome in cytogenetically normal acute myeloid leukemia. N Engl J Med 2008; 358: 1909–1918.
Wouters BJ, Lowenberg B, Erpelinck-Verschueren CA, van Putten WL, Valk PJ, Delwel R . Double CEBPA mutations, but not single CEBPA mutations, define a subgroup of acute myeloid leukemia with a distinctive gene expression profile that is uniquely associated with a favorable outcome. Blood 2009; 113: 3088–3091.
Pabst T, Eyholzer M, Fos J, Mueller BU . Heterogeneity within AML with CEBPA mutations; only CEBPA double mutations, but not single CEBPA mutations are associated with favourable prognosis. Br J Cancer 2009; 100: 1343–1346.
Hou HA, Lin LI, Chen CY, Tien HF . Reply to ‘Heterogeneity within AML with CEBPA mutations; only CEBPA double mutations, but not single CEBPA mutations are associated with favorable prognosis’. Br J Cancer 2009; 101: 738–740.
Zhang DE, Zhang P, Wang ND, Hetherington CJ, Darlington GJ, Tenen DG . Absence of granulocyte colony-stimulating factor signaling and neutrophil development in CCAAT enhancer binding protein alpha-deficient mice. Proc Natl Acad Sci USA 1997; 94: 569–574.
Pabst T, Mueller BU, Harakawa N, Schoch C, Haferlach T, Behre G et al. AML1-ETO downregulates the granulocytic differentiation factor C/EBPalpha in t(8;21) myeloid leukemia. Nat Med 2001; 7: 444–451.
Helbling D, Mueller BU, Timchenko NA, Hagemeijer A, Jotterand M, Meyer-Monard S et al. The leukemic fusion gene AML1-MDS1-EVI1 suppresses CEBPA in acute myeloid leukemia by activation of calreticulin. Proc Natl Acad Sci USA 2004; 101: 13312–13317.
Helbling D, Mueller BU, Timchenko NA, Schardt J, Eyer M, Betts DR et al. CBFB-SMMHC is correlated with increased calreticulin expression and suppresses the granulocytic differentiation factor CEBPA in AML with inv(16). Blood 2005; 106: 1369–1375.
Perrotti D, Cesi V, Trotta R, Guerzoni C, Santilli G, Campbell K et al. BCR-ABL suppresses C/EBPalpha expression through inhibitory action of hnRNP E2. Nat Genet 2002; 30: 48–58.
Radomska HS, Basseres DS, Zheng R, Zhang P, Dayaram T, Yamamoto Y et al. Block of C/EBP alpha function by phosphorylation in acute myeloid leukemia with FLT3 activating mutations. J Exp Med 2006; 203: 371–381.
Kumagai T, Akagi T, Desmond JC, Kawamata N, Gery S, Imai Y et al. Epigenetic regulation and molecular characterization of C/EBPalpha in pancreatic cancer cells. Int J Cancer 2009; 124: 827–833.
Chim CS, Wong AS, Kwong YL . Infrequent hypermethylation of CEBPA promotor in acute myeloid leukaemia. Br J Haematol 2002; 119: 988–990.
Wouters BJ, Jorda MA, Keeshan K, Louwers I, Erpelinck-Verschueren CA, Tielemans D et al. Distinct gene expression profiles of acute myeloid/T-lymphoid leukemia with silenced CEBPA and mutations in NOTCH1. Blood 2007; 110: 3706–3714.
Tien HF, Wang CH, Lin MT, Lee FY, Liu MC, Chuang SM et al. Correlation of cytogenetic results with immunophenotype, genotype, clinical features, and ras mutation in acute myeloid leukemia. A study of 235 Chinese patients in Taiwan. Cancer Genet Cytogenet 1995; 84: 60–68.
An International System for Human Cytogenetic Nomenclature (1985) ISCN 1985. Report of the Standing Committee on Human Cytogenetic Nomenclature. Birth Defects Orig Artic Ser 1985; 21: 1–117.
Hou HA, Chou WC, Lin LI, Chen CY, Tang JL, Tseng MH et al. Characterization of acute myeloid leukemia with PTPN11 mutation: the mutation is closely associated with NPM1 mutation but inversely related to FLT3/ITD. Leukemia 2008; 22: 1075–1078.
Tang JL, Hou HA, Chen CY, Liu CY, Chou WC, Tseng MH et al. AML1/RUNX1 mutations in 470 adult patients with de novo acute myeloid leukemia: prognostic implication and interaction with other gene alterations. Blood 2009; 114: 5352–5361.
Chou WC, Tang JL, Lin LI, Yao M, Tsay W, Chen CY et al. Nucleophosmin mutations in de novo acute myeloid leukemia: the age-dependent incidences and the stability during disease evolution. Cancer Res 2006; 66: 3310–3316.
Lin LI, Lin TC, Chou WC, Tang JL, Lin DT, Tien HF . A novel fluorescence-based multiplex PCR assay for rapid simultaneous detection of CEBPA mutations and NPM mutations in patients with acute myeloid leukemias. Leukemia 2006; 20: 1899–1903.
Boissel N, Renneville A, Biggio V, Philippe N, Thomas X, Cayuela JM et al. Prevalence, clinical profile, and prognosis of NPM mutations in AML with normal karyotype. Blood 2005; 106: 3618–3620.
Müller C, Bremer A, Schreiber S, Eichwald S, Calkhoven CF . Nucleolar retention of a translational C/EBPalpha isoform stimulates rDNA transcription and cell size. EMBO J 2010; 29: 897–909.
Baer MR, Stewart CC, Lawrence D, Arthur DC, Byrd JC, Davey FR et al. Expression of the neural cell adhesion molecule CD56 is associated with short remission duration and survival in acute myeloid leukemia with t(8;21)(q22;q22). Blood 1997; 90: 1643–1648.
Raspadori D, Damiani D, Lenoci M, Rondelli D, Testoni N, Nardi G et al. CD56 antigenic expression in acute myeloid leukemia identifies patients with poor clinical prognosis. Leukemia 2001; 15: 1161–1164.
Di Bona E, Sartori R, Zambello R, Guercini N, Madeo D, Rodeghiero F . Prognostic significance of CD56 antigen expression in acute myeloid leukemia. Haematologica 2002; 87: 250–256.
Webber BA, Cushing MM, Li S . Prognostic significance of flow cytometric immunophenotyping in acute myeloid leukemia. Int J Clin Exp Pathol 2008; 1: 124–133.
Villa R, Morey L, Raker VA, Buschbeck M, Gutierrez A, De Santis F et al. The methyl-CpG binding protein MBD1 is required for PML-RARalpha function. Proc Natl Acad Sci USA 2006; 103: 1400–1405.
Grignani F, De Matteis S, Nervi C, Tomassoni L, Gelmetti V, Cioce M et al. Fusion proteins of the retinoic acid receptor-alpha recruit histone deacetylase in promyelocytic leukaemia. Nature 1998; 391: 815–818.
Di Croce L, Raker VA, Corsaro M, Fazi F, Fanelli M, Faretta M et al. Methyltransferase recruitment and DNA hypermethylation of target promoters by an oncogenic transcription factor. Science 2002; 295: 1079–1082.
Truong BT, Lee YJ, Lodie TA, Park DJ, Perrotti D, Watanabe N et al. CCAAT/enhancer binding proteins repress the leukemic phenotype of acute myeloid leukemia. Blood 2003; 101: 1141–1148.
Lee YJ, Jones LC, Timchenko NA, Perrotti D, Tenen DG, Kogan SC . CCAAT/enhancer binding proteins alpha and epsilon cooperate with all-trans retinoic acid in therapy but differ in their antileukemic activities. Blood 2006; 108: 2416–2419.
Figueroa ME, Wouters BJ, Skrabanek L, Glass J, Li Y, Erpelinck-Verschueren CA et al. Genome-wide epigenetic analysis delineates a biologically distinct immature acute leukemia with myeloid/T-lymphoid features. Blood 2009; 113: 2795–2804.
Acknowledgements
We thank the Department of Medical Research for sequencing reaction. We also thank the Department of Laboratory Medicine for immunophenotyping and the Department of Pathology for cytogenetic analysis. Finally, we thank the faculty and staff in the Division of Hematology at the National Taiwan University Hospital (NTUH) for referring their AML patients to participate in this study. This work was supported by a research grants from the National Science Council (NSC 96-2320-B-002-039-MY3), Taiwan, and a research grant from the National Taiwan University Hospital (NTUH 98-S1052), Taiwan.
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LIL and HFT designed this research; TCL, DLO and SLY performed the laboratory research; WCC, HAH and HFT collected patient samples and clinical information; TCL, HAH and LIL analyzed and interpreted the data; and TCL, LIL and HFT wrote this manuscript.
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Lin, TC., Hou, HA., Chou, WC. et al. CEBPA methylation as a prognostic biomarker in patients with de novo acute myeloid leukemia. Leukemia 25, 32–40 (2011). https://doi.org/10.1038/leu.2010.222
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DOI: https://doi.org/10.1038/leu.2010.222
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