Fig. 4

GPX4 knockdown blunts antitumor effects of an ICI treatment regimen in obesity-related TNBC. (A) Hallmark gene sets commonly enriched in metM-Wnt^lung tumors of control mice vs. DIO mice determined by gene set enrichment analysis (GSEA) (n = 12–13/group). (B) Heat map of selected immune related genes in metM-Wnt^lung tumors from DIO mice, relative to controls (n = 13, normalized to gene expression from n = 12 tumors from controls) (C) Study design to assess the potential of GPX4 expression and immunotherapy treatment to ameliorate metM-Wnt^lung tumor growth in a murine model of TNBC and DIO. (D and E) Body weights (D) and tumor weights (E) of DIO mice receiving orthotopic injection of metM-Wnt^lung shScramble or shGPX4 cells, combined with either ICI or IgG treatment. Measurements taken at study endpoint (n = 15/group). (F and G) Percent tumoral CD8+ (F) and CD4+ (G) cells in tumors, as a percentage of CD45 + cells (n = 8/group). (H) Percent tumoral PD1+/TIM3 + exhausted CD8 + T cells, as a percentage of CD3 + T cells (n = 8/group). (I) Percent tumoral FoxP3 + regulatory T cells as a percentage of CD4 + cells (n = 8/group). (J) Percent tumoral putative MDSCs, as a percentage of CD45 + cells (n = 8/group). Statistical significance determined by one-way ANOVA with Tukey’s multiple comparison test (D), two-way ANOVA with Tukey’s multiple comparison test (E-J) (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001)