FIGURE 5 from EZH2 Inhibition Promotes Tumor Immunogenicity in Lung Squamous Cell Carcinomas

EZH2 inhibition alone and combined with immunotherapy is effective at controlling tumor burden in mouse models of LSCC. A, Representative MRI scans of autochthonous mice from each treatment arm at baseline and after treatment. B, Waterfall plot showing change in tumor volume for each mouse on all treatment arms, ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05 by one-way ANOVA with multiple comparisons and Holm-Šídák post hoc test on log₂-transformed values. C, H&E and HALO nuclear phenotyper images showing the cells within an autochthonous Lkb1/Pten tumor and a syngeneic graft seeded from Lkb1/Pten tumoroids. D, Percentage tumor growth from the syngeneic mouse model during 14 days of indicated treatments. ***, P = 0.0004; ****, P < 0.0001 by one-way ANOVA with multiple comparisons and Holm-Šídák post hoc test, *, P = 0.024 by two-tailed t test on log₂-transformed values, Mice/tumors n are placebo = 4/8, EPZ6438 = 5/9, anti-PD1 = 6/8, combo = 5/9, mean ± SEM. is plotted. E, Flow cytometry analysis of dissociated tumors from the syngeneic grafts from the indicated treatment arms at day 14. Percentage of EpCAM+ cells expressing IA/IE or PD-L1 are graphed, mean ± SEM is plotted, placebo n = 7, EZH2 inhibitor n = 7, anti-PD1 n = 8, combo n = 7 with two experimental replicates each, *, P = 0.035; ***, P = 0.0008 by one-way ANOVA with multiple comparisons and Holm-Šídák post hoc test. F, From the same tumor grafts, MFI for HLA-A in the EpCAM+ cells was graphed, mean ± SEM is plotted, placebo n = 7, EZH2 inhibitor n = 7, anti-PD1 n = 8, combo n = 7 with 2 experimental replicates each, **, P = 0.0015 by one-way ANOVA with multiple comparisons and Holm-Šídák post hoc test. G, From tumor grafts, PD1+/CD3+/CD4+ cells and PD1+/CD3+/CD8+ were gated and percentage of cells bound to Rat-IgG2A antibody are graphed, mean ± SEM is plotted, placebo n = 6, EZH2 inhibitor n = 6, anti-PD1 n = 7, combo n = 7; **, P < 0.006; ***, P = 0.0001; ****, P < 0.0001 by one-way ANOVA with multiple comparisons and Holm-Šídák post hoc test. H, From the grafts, percentage of CD3+/SSC-low cells within the CD45+ fraction and percentage of CD8+ cells within the CD3+ fraction were graphed, please see Supplementary Fig. S5C for representative gates, placebo n = 8, EZH2 inhibitor n = 8, anti-PD1 n = 9, combo n = 7 with two experimental replicates each, *, P = 0.0197; ****, P < 0.0001 by one-way ANOVA with multiple comparisons and Holm-Šídák post hoc test. See also Supplementary Fig. S5.