The effect of decitabine on megakaryocyte maturation and platelet releaseFinancial support:This work was supported by NIH/NIDA grants DA020816 and DA004315.
Received: 22 November 2010
Accepted after major revision: 03 May 2011
25 November 2017 (online)
Thrombocytopenia is a common feature of myelodysplastic syndromes (MDS). 5-aza-2’-deoxycytidine (decitabine) has been used to treat MDS with an approximately 20% response rate in thrombocytopenia. However, the mechanism of how decitabine increases platelet count is not clear. In this study, we investigated the effect of decitabine on megakaryocyte maturation and platelet release in the mouse. The effect of decitabine on megakaryocyte maturation was studied in an in vitro megakaryocyte differentiation model utilising mouse bone marrow cells and mouse megakaryoblastic cell line L8057. Decitabine (2.5 μM) is able to induce L8057 cells to differentiate into a megakaryocyte-like polyploidy cells with positive markers of acetylcholinesterase and αIIb integrin (CD41). Higher expression of αIIb integrin was also found in primary mouse bone marrow cells and human cord blood CD34+ cells cultured with both thrombopoietin and decitabine as compared to thrombopoietin alone. In addition, we noted a 30% platelet count increase in Balb/c mice 12 hours after the injection of decitabine at a clinically relevant dose (15 mg/m2), suggesting a rapid platelet release from the spleen or bone marrow. Our data suggest that decitabine increases platelet counts by enhancing platelet release and megakaryocyte maturation.
- 1 Kantarjian H, Giles F, List A. et al. The incidence and impact of thrombocytopenia in myelodysplastic syndromes. Cancer 2007; 109: 1705-1714.
- 2 Steensma DP. Myelodysplasia, megakaryocytes, and methylation. Leuk Res 2004; 28: 775-776.
- 3 Nolte F, Hofmann WK. Molecular mechanisms involved in the progression of myelodysplastic syndrome. Future Oncol 2010; 6: 445-455.
- 4 Lee ST, Jang JH, Suh HC. et al. Idarubicin, cytarabine, and topotecan in patients with refractory or relapsed acute myelogenous leukemia and high-risk myelodysplastic syndrome. Am J Hematol 2001; 68: 237-245.
- 5 List A, Kurtin S, Roe DJ. et al. Efficacy of lenalidomide in myelodysplastic syndromes. N Engl J Med 2005; 352: 549-557.
- 6 Rosenfeld CS, Zeigler ZR, Shadduck RK. et al. Phase II study of roquinimex in myelodysplastic syndrome. Am J Clin Oncol 1997; 20: 189-192.
- 7 Platzbecker U, Haase M, Herbst R. et al. Activity of sirolimus in patients with myelodysplastic syndrome–results of a pilot study. Br J Haematol 2005; 128: 625-630.
- 8 Saba HI. Decitabine in the treatment of myelodysplastic syndromes. Ther Clin Risk Manag 2007; 3: 807-817.
- 9 de Vos D, van Overveld W. Decitabine: a historical review of the development of an epigenetic drug. Ann Hematol 2005; 84 (Suppl. 01) 3-8.
- 10 Issa JP. Decitabine. Curr Opin Oncol 2003; 15: 446-451.
- 11 Bonifer C, Lefevre P, Tagoh H. The regulation of chromatin and DNA-methylation patterns in blood cell development. Curr Top Microbiol Immunol 2006; 310: 1-12.
- 12 Hennessy BT, Garcia-Manero G, Kantarjian HM. et al. DNA methylation in haematological malignancies: the role of decitabine. Expert Opin Investig Drugs 2003; 12: 1985-1993.
- 13 Galm O, Herman JG, Baylin SB. The fundamental role of epigenetics in hematopoietic malignancies. Blood Rev 2006; 20: 1-13.
- 14 Gao S, Skeldal S, Krogdahl A. et al. CpG methylation of the PAI-1 gene 5’-flanking region is inversely correlated with PAI-1 mRNA levels in human cell lines. Thromb Haemost 2005; 94: 651-660.
- 15 Saunthararajah Y, Hillery CA, Lavelle D. et al. Effects of 5-aza-2’-deoxycytidine on fetal hemoglobin levels, red cell adhesion, and hematopoietic differentiation in patients with sickle cell disease. Blood 2003; 102: 3865-3870.
- 16 Pecci A, Malara A, Badalucco S. et al. Megakaryocytes of patients with MYH9-related thrombocytopenia present an altered proplatelet formation. Thromb Haemost 2009; 102: 90-96.
- 17 van den Oudenrijn S, von dem Borne AE, de Haas M. Differences in megakaryocyte expansion potential between CD34(+) stem cells derived from cord blood, peripheral blood, and bone marrow from adults and children. Exp Hematol 2000; 28: 1054-1061.
- 18 Herman JG, Graff JR, Myohanen S. et al. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA 1996; 93: 9821-9826.
- 19 Oakeley EJ, Schmitt F, Jost JP. Quantification of 5-methylcytosine in DNA by the chloroacetaldehyde reaction. Biotechniques 1999; 27: 744-752.
- 20 Kaushansky K. Thrombopoietin: the primary regulator of megakaryocyte and platelet production. Thromb Haemost 1995; 74: 521-525.
- 21 Vainchenker W, Methia N, Debili N. et al. c-mpl, the thrombopoietin receptor. Thromb Haemost 1995; 74: 526-528.
- 22 Wadhwa M, Thorpe R. Haematopoietic growth factors and their therapeutic use. Thromb Haemost 2008; 99: 863-873.
- 23 Teofili L, Martini M, Di Mario A. et al. Expression of p15(ink4b) gene during megakaryocytic differentiation of normal and myelodysplastic hematopoietic progenitors. Blood 2001; 98: 495-497.
- 24 Kanaji S, Kanaji T, Jacquelin B. et al. Thrombopoietin initiates demethylation-based transcription of GP6 during megakaryocyte differentiation. Blood 2005; 105: 3888-3892.
- 25 Gandhi MJ, Drachman JG, Reems JA. et al. A novel strategy for generating platelet-like fragments from megakaryocytic cell lines and human progenitor cells. Blood Cells Mol Dis 2005; 35: 70-73.
- 26 Kaminska J, Klimczak-Jajor E, Skierski J. et al. Effects of inhibitor of Src kinases, SU6656, on differentiation of megakaryocytic progenitors and activity of alpha1,6-fucosyltransferase. Acta Biochim Pol 2008; 55: 499-506.
- 27 Lannutti BJ, Blake N, Gandhi MJ. et al. Induction of polyploidization in leukemic cell lines and primary bone marrow by Src kinase inhibitor SU6656. Blood 2005; 105: 3875-3878.
- 28 Bluteau D, Lordier L, Di Stefano A. et al. Regulation of megakaryocyte maturation and platelet formation. J Thromb Haemost 2009; 7 (Suppl. 01) 227-234.
- 29 van den Bosch J, Lubbert M, Verhoef G. et al. The effects of 5-aza-2’-deoxycytidine (Decitabine) on the platelet count in patients with intermediate and high-risk myelodysplastic syndromes. Leuk Res 2004; 28: 785-790.
- 30 Hofmann WK, Kalina U, Koschmieder S. et al. Defective megakaryocytic development in myelodysplastic syndromes. Leuk Lymphoma 2000; 38: 13-19.
- 31 Kalina U, Hofmann WK, Koschmieder S. et al. Alteration of c-mpl-mediated signal transduction in CD34(+) cells from patients with myelodysplastic syndromes. Exp Hematol 2000; 28: 1158-1163.