Figure 5 from Metabolic Reprogramming of Tumor-Associated Macrophages Using Glutamine Antagonist JHU083 Drives Tumor Immunity in Myeloid-Rich Prostate and Bladder Cancers

JHU083-induced glutamine antagonism affects tumor cell metabolism and induces cell death in urologic tumors. A, Log-fold changes of glutamine utilizing enzymes after JHU083 treatment vs. control tumors in CD45⁻ sorted cells from B6CaP tumors, followed by scRNA-seq. B, Western blot showing qualitative changes in the levels of glutamine synthesizing/utilizing enzymes and transporters in the CD45⁻ fraction of MB49 tumors. C, Percentage of GLUT1⁺ CD45⁻ live cells determined by flow cytometry in B6CaP tumors (n = 7/group). D, Targeted metabolomic analysis of B6CaP tumors by LC-MS/MS (n = 3/group). E, Volcano plot showing key metabolite levels of JHU083-treated vs. nontreated control tumors based on the metabolomic analysis shown in D. F, Absolute quantification of metabolites by LC/MS-MS (n = 3 or 5/group). G and H, Western blot images showing qualitative changes in c-MYC, phospho-c-MYC, and HIF-1ɑ in MB49 tumors following JHU083 treatment, and (H and I) MTT assay in DON-treated MB49 cells and immunoblot of cleaved caspase 3 quantification in CD45⁻ fraction MB49 tumors (J). Statistical analyses were performed using the unpaired t test. (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001).