Extended Data Fig. 1: Kras activation drives metabolic reprogramming and confers resistance to GLS inhibition.
From: The amino acid transporter SLC7A5 is required for efficient growth of KRAS-mutant colorectal cancer
a, Normalized enrichment scores (NES) of metabolic signature gene sets from ‘Hallmark’ gene set collection in Apcfl/fl KrasG12D/+ mouse intestine. Gene sets with nominal P values after multiple hypotheses correction and FDR, q < 0.05 indicated for each gene set. b, Diagram of glycolysis and glutaminolysis pathway intermediates. 13C-glucose (13C6)-labelled carbons (red); 13C-Glutamine (13C5)-labelled carbons (blue). c, Schematic illustrating role of GLS in glutaminolysis. d, Glutamine hydrolysis in Apcfl/fl KrasG12D/+ organoids after 24 h treatment with GLS inhibitor (GLSi; 1 µM), CB839 (1 µM), BPTES (20 µM), or DMSO (vehicle) followed by 13C5-Glutamine–tracing using LC-MS. Plots show mean percentage of 13C5-glutamine-derived 13C5-glutamate (peak area/µg of protein) ± s.e.m. Each treatment performed as an independent experiment. *P =0.05, one-way Mann–Whitney U test. e, Relative viability (mean ± s.e.m.) of Apcfl/fl KrasG12D/+ organoids treated as in d for 72 h. f, H&E and BrdU staining showing resistance to GLSi treatment for 72 h in the small intestine (SI) and colon of Apcfl/fl KrasG12D/+ mice (representative of 4 biologically independent mice). Treatment began 24 h after induction. Scale bars, 100 µm. g, Number of BrdU-positive cells per half-crypt (mean ± s.e.m.) in SI and colon from (f). n=4 biological replicates per group; 25 half-crypts were scored per mouse. h, Schematic illustrating leucine or valine transamination by BCAT and nitrogen (orange ovals) fate after transamination. i, Plots show mean percentage of 15N-leucine-derived or 15N-valine-derived glutamate (peak area/µg of protein) ± s.e.m. j, Mass spectrometry imaging (MSI) of 13C5-glutamine derived TCA cycle intermediates in APC and APC KRAS small intestinal tissue. Plots show ion abundance of isotope-labelled glutamine-derived intermediates ± s.e.m. n = 4 biological replicates per group. P using two-way ANOVA, with Tukey’s multiple comparison test. In d, e, i n = 3 biological replicates in technical triplicates each per group.
