Figure 1 from Metabolic Reprogramming of Tumor-Associated Macrophages Using Glutamine Antagonist JHU083 Drives Tumor Immunity in Myeloid-Rich Prostate and Bladder Cancers
Authors: Praharaj Monali , Shen Fan , Lee Alex J. , Zhao Liang , Nirschl Thomas R. , Theodros Debebe , Singh Alok K. , Wang Xiaoxu , Adusei Kenneth M. , Lombardo Kara A. , Williams Raekwon A. , Sena Laura A. , Thompson Elizabeth A. , Tam Ada , Yegnasubramanian Srinivasan , Pearce Edward J. , Leone Robert D. , Alt Jesse , Rais Rana , Slusher Barbara S. , Pardoll Drew M. , Powell Jonathan D. , Zarif Jelani C. 
Published: July 2024

<p>The antitumor activity of the glutamine antagonist JHU083 against urologic tumors is myeloid dependent. <b>A,</b> Dot plots showing expression levels and fractional abundance of myeloid cells expressing glutamine metabolism enzymes, <i>GLUL,</i> and <i>GLS</i> across samples (tumor, involved, distal, and benign). <b>B,</b> Violin plots showing the expression levels and fractional abundance of different cell types expressing <i>GLUL</i> and <i>GLS</i>. Data for <b>A</b> and <b>B</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 (<a href="#bib41" target="_blank">41</a>). <b>C,</b> Schematic diagrams showing the heterotopic syngeneic urologic tumor (B6CaP and MB49) models and therapeutic JHU083 treatment strategy. <b>D</b> and <b>E,</b> Tumor volume and tumor weight measurements (post-necropsy) in the B6CaP (<i>n</i> = 8/group) and MB49 (<i>n</i> = 9 or 10/group) tumors. <b>F</b> and <b>G,</b> Effects of CD8<sup>+</sup> T cell depletion on tumor volume and animal survival for (<b>F</b>) MB49 (<i>n</i> = 4–5/group) and (<b>G</b>) B6CaP (<i>n</i> = 4–6/group) tumors, respectively. <b>H</b> and <b>I,</b> Effect of CD4<sup>+</sup> T cell depletion on tumor volume of (<b>H</b>) MB49 (<i>n</i> = 4–5/group) and (<b>I</b>) B6CaP (<i>n</i> = 3–5/group) tumors and animal survival. Antitumor activity of JHU083-reprogrammed and adoptively transferred (<b>J)</b> TAMs (CD45<sup>+</sup> Ly6G<sup>−</sup> CD3<sup>−</sup> CD11b<sup>+</sup> F4.80<sup>+</sup>; <i>n</i> = 5 or 10/group) and (<b>K</b>) TIMs (CD45<sup>+</sup> Ly6G<sup>−</sup> CD3<sup>−</sup> Ly6C <sup>(hi)</sup>; <i>n</i> = 4 or 7/group) on MB49 tumor volume (<b>L</b>). 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 <i>t</i> test or a two-way ANOVA using Bonferroni’s multiple comparisons (*, <i>P</i> < 0.05; **, <i>P</i> < 0.01; ***, <i>P</i> < 0.001; ****, <i>P</i> < 0.0001). Log-rank (Mantel–Cox) tests were performed for survival analysis (<b>F,</b> JHU083 relative to control, <i>P</i> = 0.0412; α-CD8b relative to α-CD8b + JHU083, <i>P</i> = 0.0342; α-CD8b + JHU083 vs. JHU083, <i>P</i> = ns. <b>I,</b> JHU083 vs. control, <i>P</i> = 0.01; α-CD4 vs. α-CD4 + JHU083, <i>P</i> = 0.0801; α-CD4 + JHU083 vs. JHU083, <i>P</i> = ns).</p>