Expression data from AZA treated megakaryocytic cells under TPO-RA stimulation

Thrombocytopenia is an independent prognostic indicator in patients with MDS (de Swart, L. et al, Br J Haematol, 2015) and we sought to evaluate the contribution of azacitidine to the worsening of this condition in patients with MDS and acute myeloid leukemia (AML). Since MDS/AML may have lower amounts of residual healthy cells, we sought to model this effect of AZA on healthy cord blood derived megakaryocytic cells. We used microarrays to detail the global programme of gene expression in the presence of azacitidine under megakaryocytic differentiation stimulants and identified distinct classes of dysregulated genes and pathways during this process. To assess the effects of AZA on megakaryocytic cells at the transcriptional level, we treated cord blood mononuclear cells with or without 0.3uM AZA for 16 hours in serum and cytokine free conditions in plain stem span media in the presence of thrombopoietin receptor agonists (TPO-RA) 100ng/mL TPO or 5ug/mL eltrombopag (EP). Followed by FACS sorting of viable cMPL expressing cells for microarray analysis.