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. Author manuscript; available in PMC: 2018 Apr 2.
Published in final edited form as: Nat Med. 2017 Oct 2;23(11):1362–1368. doi: 10.1038/nm.4407

Figure 4. Keap1-mutant cells display a robust sensitivity to glutaminase inhibition.

Figure 4

a) Schematic of glutamine uptake by Slc1a5 and hydrolysis of glutamine to glutamate by Gls. Inhibitors of Gls are shown in red. b) Relative viability assayed by cell-titer glo (relative luminescent units) on KP and KPK cells after treatment with CB-839 (left) or BPTES (right) for 72 hrs. All data points are relative to vehicle treated controls (n = 4 technical replicates/data point). c) Cumulative population doublings of KP and KPK cells in the presence of vehicle, CB-839 or BPTES (n = 4 technical replicates/data point) after 6 days in culture. d) Trypan blue exclusion viability counts of indicated human lung cancer cell lines. Each cell line was cultured in the presence of vehicle or 500nM CB-839 (n = 4 technical replicates/cell line). Displayed results are normalized against vehicle treated cell lines after 72 hrs of treatment. A549 and H1975 are TP53-wild type, all others are TP53-mutant. e) Subcutaneous tumor volumes of KP and KPK treated with vehicle or CB-839 starting from day 13 measured over time for 25 days (n = 6 tumors/genotype/treatment). Related to Fig 4f. f) Final tumor masses related to Supplementary Data Fig 11b. *p < 0.05, ****p < 0.0001 obtained from 1-way ANOVA with Tukey's post hoc test. g) Orthotopic growth measurements of KP and KPK cells treated with vehicle or CB-839 starting from day 13 (n = 4 mice/genotype/treatment). Quantitation of luminescence (photon flux) in mice orthotopically transplanted with KP or KPK cells transduced with a vector expressing Luciferase. Relative photon flux calculated by normalizing all time points per animal to initial measurements at 10 days post transplantation. Individual groups depicted in Supplementary Data Fig 11c. ***p < 0.001 obtained from 2-way ANOVA. h) Subcutaneous tumor volumes of KP-ix (inducible GOF-Nrf2) treated with vehicle or CB-839 in the presence or absence of doxycycline (DOX) (n = 6 mice/DOX treatment). Individual groups and full experiment depicted in Supplementary Data Fig 11d. i) Five patient-derived xenograft (PDX) models treated with vehicle or CB-839 for the indicated amount of days. Individual groups and full experiments depicted in Supplementary Data Fig 11g and h. All error bars depict s.e.m.