Figure 4 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 caused a divergent metabolic response affecting glycolysis, purine metabolism, and succinate in TAMs. A and B, Surface and intracellular expression of GLUT1 and HKII (percentage positive population and mean fluorescence intensity) on B6CaP-derived TAMs. C, GSEA showing the enrichment of the Hallmark glycolysis gene set among DEGs in bulk RNA-sequenced FACS-sorted B6CaP-derived TAMs following JHU083 treatment relative to control. D, Heat map showing the differential metabolites in TAMs sorted from JHU083-treated and control B6CaP tumors (n = 3/group). E, Volcano plot showing log2 fold change vs. –log10 (FDR-corrected P value), representing fold changes in metabolite abundance in one-carbon metabolism, purine nucleotide metabolism, and hexosamine pathway in JHU083-treated vs. control TAMs. F, Normalized relative labeled metabolites from U-13C glucose in the TCA cycle in TAMs derived from B6CaP tumors (n = 9/group in two independent experiments). G, Normalized relative metabolite abundances in the TCA cycle in TAMs derived from B6CaP tumors (n = 9/group from two independent experiments) and (H) log fold-change of TCA cycle enzymes and inflammatory cytokine transcripts in TAMs (from scRNA-seq at an early time point (day 7 posttreatment). DEGs for GSEA in C were calculated with DESeq2, and statistics on genes of interest in H were calculated with the Wilcoxon rank-sum test. All other statistical analyses were performed using the unpaired t test (*, P P P P