Figure 1 from Metabolic Reprogramming of Tumor-Associated Macrophages Using Glutamine Antagonist JHU083 Drives Tumor Immunity in Myeloid-Rich Prostate and Bladder Cancers

The antitumor activity of the glutamine antagonist JHU083 against urologic tumors is myeloid dependent. A, Dot plots showing expression levels and fractional abundance of myeloid cells expressing glutamine metabolism enzymes, GLUL, and GLS across samples (tumor, involved, distal, and benign). B, Violin plots showing the expression levels and fractional abundance of different cell types expressing GLUL and GLS. Data for A and B were generated using the previously published and publicly available scRNA-seq integrated dataset from nine metastatic prostate cancer and seven benign BM control patient samples by Kfoury and colleagues (41). C, Schematic diagrams showing the heterotopic syngeneic urologic tumor (B6CaP and MB49) models and therapeutic JHU083 treatment strategy. D and E, Tumor volume and tumor weight measurements (post-necropsy) in the B6CaP (n = 8/group) and MB49 (n = 9 or 10/group) tumors. F and G, Effects of CD8+ T cell depletion on tumor volume and animal survival for (F) MB49 (n = 4–5/group) and (G) B6CaP (n = 4–6/group) tumors, respectively. H and I, Effect of CD4+ T cell depletion on tumor volume of (H) MB49 (n = 4–5/group) and (I) B6CaP (n = 3–5/group) tumors and animal survival. Antitumor activity of JHU083-reprogrammed and adoptively transferred (J) TAMs (CD45+ Ly6G− CD3− CD11b+ F4.80+; n = 5 or 10/group) and (K) TIMs (CD45+ Ly6G− CD3− Ly6C (hi); n = 4 or 7/group) on MB49 tumor volume (L). Briefly, live, FACS-sorted TAMs or TIMs were mixed in a 1:1 ratio with cultured MB49 cells and were subsequently implanted on flanks to generate tumors in recipient mice. Data are represented as mean values ± SEM. All experiments were independently validated, at least in two separate biological replicates. Statistical analyses were performed using either t test or a two-way ANOVA using Bonferroni’s multiple comparisons (*, P P P P F, JHU083 relative to control, P = 0.0412; α-CD8b relative to α-CD8b + JHU083, P = 0.0342; α-CD8b + JHU083 vs. JHU083, P = ns. I, JHU083 vs. control, P = 0.01; α-CD4 vs. α-CD4 + JHU083, P = 0.0801; α-CD4 + JHU083 vs. JHU083, P = ns).