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CUDC-907 monotherapy in AML
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Figure 2. CUDC-907 treatment induces apoptosis and inhibits colony formation in primary acute myeloid leukemia cells, but spares normal human bone marrow mononuclear cells. (A) Primary samples from patients with FLT3-ITD-positive and FLT3-ITD-negative acute myeloid leukemia (AML) (n=14 and n=61, respectively) were treated with variable concentrations of CUDC-907 in 96-well plates for 72 h and then viable cells were determined using MTT reagent. The IC50 values are means of dupli- cates from one experiment due to limited samples. The horizontal lines indicate the median. (B, C) Primary AML patient samples were treated with CUDC-907 for 24 h and then subjected to annexin V-FITC/propidium iodide (PI) staining and flow cytometry analyses. Representative dot plots are shown (B). Mean percent of annexin V+ cells ± standard error of mean (SEM) are shown (panel C). (D) Primary AML patient samples were treated with CUDC-907 for 24 h. Whole cell lysates were subjected to western blotting. (E) Primary AML patient samples were cultured with vehicle control or 100 nM CUDC-907 for 24 h and then plated in methylcellulose. After incubation for 2 weeks, the number of surviving AML cells capable of generating leukemia colonies (AML-CFU) were enumerated. Data are presented as mean ± SEM. **P<0.01, ***P<0.001. (F) Normal human bone marrow mononuclear cells from a single donor were cultured with vehicle control or 100 nM CUDC-907 for 24 h and then plated in methylcellulose. After incubation for 2 weeks, the number of surviving hematopoietic cells capable of generating colonies was counted. Total erythroid and myeloid colonies are presented as mean ± SEM. (G) The numbers of BFU-E, CFU-E, CFU-G, CFU-M, CFU-GM, and CFU-GEMM colonies are presented as mean ± SEM. AML: acute myeloid leukemia; CUDC: CUDC-907; cf-Caspase 3: cleaved caspase 3; cf-PARP: cleaved PARP; BMMNC: bone marrow mononuclear cells; BFU-E: burst-forming unit – erythroid; CFU-E: colony-form- ing unit - erythroid, CFU-G: colony-forming unit – granulocyte; CFU-M: colony-forming unit - monocyte, CFU-GM: colony-forming unit – granulocyte, macrophage; CFU-GEMM: colony-forming unit – granulocyte, erythroid, macrophage megaryocyte.
haematologica | 2019; 104(11)
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