Fig. 4: Blockade of glutamine pathway through glutaminase knockdown and glutaminase inhibition in combination with complex I inhibition induces metabolic shutdown in T-ALL cells in vitro. | Nature Communications

Fig. 4: Blockade of glutamine pathway through glutaminase knockdown and glutaminase inhibition in combination with complex I inhibition induces metabolic shutdown in T-ALL cells in vitro.

From: Inhibition of mitochondrial complex I reverses NOTCH1-driven metabolic reprogramming in T-cell acute lymphoblastic leukemia

Fig. 4

a Enrichment of glutamine metabolism components in T-ALL TARGET cohort; (Gene Ontology analysis); two-sided t test; The center line represents the median and whiskers represents maximum (Q3+1.5*IQR) and minimum value (Q1+1.5*IQR). b Viability of NOTCH1-mutated T-ALL cell lines (96 h, 0–100 nM of IACS01759) upon following conditions: no nutrients, pyruvate (Pyr), glutamine (Gln), glutamine+pyruvate (Glut+Pyr), glucose+pyruvate (Glc+Pyr), glucose+glutamine+pyruvate (Glc+Glut+Pyr), normalized to DMSO. c AUC upon IACS-010759 treatment (by CTG) calculated from (b), normalized to DMSO from Gln+Glc+Pyr, for NOTCH1-wt (blue) (n = 3) and NOTCH1-mutated (black) T-ALL cell lines (n = 4) (mean ± SD, n = 3 independent experiments, n = 3 replicates/condition). Two-way ANOVA; *p = 0.02; ***p = 0.0002; ****p < 0.0001; d Representative OCR response in NOTCH1-mutated T-ALL cell lines JURKAT subjected to doxycycline-induced knockdown of GLS (blue) and treatment with IACS-010759 (10 nM) for 4 h (red), or the combination (green) (mean ± SD, n = 3 biological replicates, n = 4 replicates/condition). e Cell viability in NOTCH1-mutated T-ALL patient samples (n = 4) treated with CB-839 (1 μM) and IACS-01759 (10 nM) after 96 h, by FL (mean ± SD, n = 4 independent patient samples, n = 3 replicates/condition); one-way ANOVA; *p = 0.039; **p = 0.0017; ****p < 0.0001. f Reactive oxygen species (ROS) -MFI of in NOTCH1-mutant T-ALL cell lines (n = 3) treated 96 h with CB-839 (1 μM) and IACS-010759 (10 nM). (mean ± SD, n = 3) by FL; one-way ANOVA; *p = 0.0202; ***p = 0.0005; ****p < 0.0001. g Basal OCR in NOTCH1-mutated T-ALL cell lines after treatment with vehicle, IACS-010759 (10 nM, 4 h), CB-839 (1 μM, 12 h), or IACS-010759/CB-839 combination (mean ± SD, n = 7 cell lines, n = 3 independent experiments, n = 4 replicates per condition); one-way ANOVA: ***p = 0.0003; ****p < 0.0001. h SIRM ultra-HPLC MS of NOTCH1-mutated T-ALL cell line PF-382 after enrichment with 13C515N2-glutamine and treatment with the combination of 10 nM IACS-010759 and 1 μM CB-839 or vehicle for 12 h. i Gene expression analysis in NOTCH1-mutant T-ALL after treatment with the combination of 10 nM IACS-010759 and 1 μM CB-839 compared to vehicle; (x-gene rank in the pathway list; y-running enrichment score).

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