Fig. 6: Co-targeting of PPIA and glutaminase potently inhibits tumor growth of NRF2-hyperactivated NSCLC.

A Cell viability of A549 cells treated with CsA (8 μM) and CB-839 (0.08 μM) combination (Com) in presence of 2, 1, 0.5 mM glutamine (Gln). B 3D clonogenic assay of A549-mCherry cells treated with vehicle, CsA, CB-839, or their combination (n = 3 independent experiments). After growing for 2 weeks, formed clones were quantitated by using Image J. The synergistic anti-proliferative effect was evaluated by Combenefit. Blue indicates synergy, while red indicates antagonism between drugs. C Lung orthotopic model was established by tail-vein injection of A549-Luc cells. Mice were divided into four groups (n = 6 mice per group): vehicle control (p.o. twice daily), CB-839 (150 mg/kg, p.o. twice daily), CsA (20 mg/kg, i.p. every 3 days), and CB-839/CsA combination (150 mg/kg, p.o. twice daily; 20 mg/kg, i.p. every 3 days). Bioluminescence imaging was performed every 7 days using a PerkinElmer IVIS Spectrum CT system. D Statistics of lung metastatic nodules in the lung orthotopic model with different treatments (n = 6 mice per group). E Kaplan–Meier survival analysis of different groups in lung orthotopic model formed by tail-vein injection of A549-luc cells (n = 6 mice per group). F, G Tumor growth in NCG mice bearing MT−101 F or WT-201 G PDX xenograft treated with the vehicle (p.o. twice daily), CB-839 (150 mg/kg, p.o. twice daily), CsA (20 mg/kg, i.p. every 3 days), and CB-839/CsA combination (n = 6 mice per group). MT−101 PDX harbors concurrent KEAP1 and KRAS mutations (KEAP1^G333C and KRAS^G12D). WT-201 PDX has KEAP1^WT and KRAS^WT. A represents mean ± SD of three independent experiments. P values were analyzed using Two-tailed unpaired Student’s t-test for A and D, using Log-rank (Mantel-Cox) test for E, and using One-way ANOVA for (F, G). P < 0.05 was considered statistically significant. Source data are provided as a Source Data file.