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posted on 2024-07-02, 07:21 authored by Monali Praharaj, Fan Shen, Alex J. Lee, Liang Zhao, Thomas R. Nirschl, Debebe Theodros, Alok K. Singh, Xiaoxu Wang, Kenneth M. Adusei, Kara A. Lombardo, Raekwon A. Williams, Laura A. Sena, Elizabeth A. Thompson, Ada Tam, Srinivasan Yegnasubramanian, Edward J. Pearce, Robert D. Leone, Jesse Alt, Rana Rais, Barbara S. Slusher, Drew M. Pardoll, Jonathan D. Powell, Jelani C. Zarif 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 < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001).
Funding
National Institutes of Health (NIH)
Prostate Cancer Foundation (PCF)
Maryland Cigarette Restitution Fund
History
ARTICLE ABSTRACT
Glutamine metabolism in tumor microenvironments critically regulates antitumor immunity. Using the glutamine-antagonist prodrug JHU083, we report potent tumor growth inhibition in urologic tumors by JHU083-reprogrammed tumor-associated macrophages (TAMs) and tumor-infiltrating monocytes. We show JHU083-mediated glutamine antagonism in tumor microenvironments induced by TNF, proinflammatory, and mTORC1 signaling in intratumoral TAM clusters. JHU083-reprogrammed TAMs also exhibited increased tumor cell phagocytosis and diminished proangiogenic capacities. In vivo inhibition of TAM glutamine consumption resulted in increased glycolysis, a broken tricarboxylic acid (TCA) cycle, and purine metabolism disruption. Although the antitumor effect of glutamine antagonism on tumor-infiltrating T cells was moderate, JHU083 promoted a stem cell–like phenotype in CD8+ T cells and decreased the abundance of regulatory T cells. Finally, JHU083 caused a global shutdown in glutamine-utilizing metabolic pathways in tumor cells, leading to reduced HIF-1α, c-MYC phosphorylation, and induction of tumor cell apoptosis, all key antitumor features. Altogether, our findings demonstrate that targeting glutamine with JHU083 led to suppressed tumor growth as well as reprogramming of immunosuppressive TAMs within prostate and bladder tumors that promoted antitumor immune responses. JHU083 can offer an effective therapeutic benefit for tumor types that are enriched in immunosuppressive TAMs.
