Fig. 1: Characterization of VEN-resistance development in the BCP-ALL cell line RS4;11. | Cell Death & Disease

Fig. 1: Characterization of VEN-resistance development in the BCP-ALL cell line RS4;11.

From: Venetoclax resistance in acute lymphoblastic leukemia is characterized by increased mitochondrial activity and can be overcome by co-targeting oxidative phosphorylation

Fig. 1

A–C BCP-ALL cell line RS4;11 was cultured and treated with increasing concentrations of VEN (ranging from 6 nM, 12 nM, 25 nM, 50 nM and 100 nM) or the corresponding concentrations of solvent DMSO over time. Five DMSO control and VEN-resistant lines were generated each over a period of 8 months of continuous treatment. To test VEN resistance development, cells were treated with increasing concentrations of VEN (DMSO, 0.1 nM; 1 nM; 10 nM; 50 nM; 100 nM; 500 nM; 1 µM; 10 µM and 25 µM) for 72 h (N = 5 biological replicates/N = 1 in technical triplicates). Determination of half maximal effective concentration values (EC50) of VENsens line 1 compared to VENins line 1-5 (FSC/SSC criteria, N = 1 in technical triplicates ± standard deviation) showed increasing EC50 values from 6.4 nM to 35.2 µM over time in all VEN treated lines. D VENsens and VENins RS4;11 lines were stained with 1 μM CellTrace™ Violet and median fluorescence intensities (MFI) were determined every 24 h from day 1 to day 7. MFI values were normalized to MFIs of day 1 (N = 3 in triplicates, ± standard deviation). E VENsens and VENins RS4;11 lines were treated with DMSO or the combination of Asparaginase, Dexamethasone and Vincristine for 72 h as stated. Cell death was assessed using FSC/SSC criteria and normalized to corresponding DMSO control. (N = 3 in triplicates). F, G VENsens and VENins RS4;11 lines were incubated with increasing concentrations of Daunorubicine (DMSO; 0.1 ng/ml; 1 ng/ml; 10 ng/ml; 50 ng/ml; 100 ng/ml; 500 ng/ml and 1 000 ng/ml) and Staurosporine (DMSO; 1 nM; 5 nM; 10 nM; 25 nM; 50 nM; 100 nM; 250 nM) for 72 h. EC50 values were determined by FSC/SSC criteria and normalized to corresponding DMSO control (N = 3 in triplicates). H VENsens and VENins RS4;11 lines were cultured under drug holiday conditions for 20 weeks. Cells were incubated with increasing concentrations of VEN (DMSO; 0.1 nM; 1 nM; 10 nM; 50 nM; 100 nM; 500 nM; 1 µM; 10 µM and 25 µM) for 72 h. Cell death was assessed using FSC/SSC criteria comparing VENsens line 1 and and VENins RS4;11 lines 1-5 (N = 1 in technical triplicates).

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