Figure 4.
MYC-dependent aerobic glycolysis is impaired in CDK7-inhibited MM cells. (A) DepMap CRISPR screen (Avana library 18Q4) dependency data indicating that MM cell lines are among the most sensitive to HK2 depletion based on cell line rank. Mean of chronos scores for each disease type are shown in the graph. (B-C) H929 and AMO1 cells were treated with DMSO or YKL-5-124 and analyzed with a glycolysis stress assay on a Seahorse XFe96 extracellular flux analyzer. (B-C) ECAR was detected at baseline, after injection of glucose, oligomycin, and 2-deoxy-D-glucos e (2-DG). Basal glycolytic rate and spare glycolytic capacity were analyzed by overall ECAR in control and YKL-5-124–treated groups at different concentrations of YKL-5-124. (D) Representative 18F-FDG PET-computed tomography images (upper panel) and quantification (lower panel) of H929 cell xenografts in mice after treatment (10 mg/kg YKL-5-124 or vehicle, for 3 days). Bar graphs represent the SUV maximum and corresponding TV for mice. (E) LDH activity was measured in cell lysate from AMO1 cells treated with YKL-5-124 for 24 hours. (F) Culture supernatant (5 μL) from both untreated cells after 6 and 24 hours of culture, and 24 hour–treated cells was used to measure lactate secretion using Lactate-Glo assay. (G) H929 and AMO1 cells were transduced with empty vector or HK2-overexpression vector and subjected to western blot analysis or treated with YKL-5-124 for 72 hours for cell killing assessment. IC50 values are shown in the graph. (H) H929 and AMO1 cells were cultured in glucose or galactose (10 mM) media for a week and treated with DMSO or increasing concentrations of YKL-5-124. Cellular viability was determined by CTG assay. (I) Three MM cell lines (H929, AMO1, and MM1S) were cultured in the presence of different concentrations of YKL-5-124 with or without bortezomib (2.5 nM), lenalidomide (5 μM), melphalan (2.5 μM), or carfilzomib (1 nM), and cell survival was assessed by CTG. Data are presented as CI values evaluated using the Calcusyn software. CI, combination index; DMSO, dimethyl sulfoxide; ECAR, extracellular acidification rate; IC50, 50% inhibitory; PET, positron emission tomography; SUV, standardized uptake value; TV, tumor volume.

MYC-dependent aerobic glycolysis is impaired in CDK7-inhibited MM cells. (A) DepMap CRISPR screen (Avana library 18Q4) dependency data indicating that MM cell lines are among the most sensitive to HK2 depletion based on cell line rank. Mean of chronos scores for each disease type are shown in the graph. (B-C) H929 and AMO1 cells were treated with DMSO or YKL-5-124 and analyzed with a glycolysis stress assay on a Seahorse XFe96 extracellular flux analyzer. (B-C) ECAR was detected at baseline, after injection of glucose, oligomycin, and 2-deoxy-D-glucos e (2-DG). Basal glycolytic rate and spare glycolytic capacity were analyzed by overall ECAR in control and YKL-5-124–treated groups at different concentrations of YKL-5-124. (D) Representative 18F-FDG PET-computed tomography images (upper panel) and quantification (lower panel) of H929 cell xenografts in mice after treatment (10 mg/kg YKL-5-124 or vehicle, for 3 days). Bar graphs represent the SUV maximum and corresponding TV for mice. (E) LDH activity was measured in cell lysate from AMO1 cells treated with YKL-5-124 for 24 hours. (F) Culture supernatant (5 μL) from both untreated cells after 6 and 24 hours of culture, and 24 hour–treated cells was used to measure lactate secretion using Lactate-Glo assay. (G) H929 and AMO1 cells were transduced with empty vector or HK2-overexpression vector and subjected to western blot analysis or treated with YKL-5-124 for 72 hours for cell killing assessment. IC50 values are shown in the graph. (H) H929 and AMO1 cells were cultured in glucose or galactose (10 mM) media for a week and treated with DMSO or increasing concentrations of YKL-5-124. Cellular viability was determined by CTG assay. (I) Three MM cell lines (H929, AMO1, and MM1S) were cultured in the presence of different concentrations of YKL-5-124 with or without bortezomib (2.5 nM), lenalidomide (5 μM), melphalan (2.5 μM), or carfilzomib (1 nM), and cell survival was assessed by CTG. Data are presented as CI values evaluated using the Calcusyn software. CI, combination index; DMSO, dimethyl sulfoxide; ECAR, extracellular acidification rate; IC50, 50% inhibitory; PET, positron emission tomography; SUV, standardized uptake value; TV, tumor volume.

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