FIGURE 5 from EZH2 Inhibition Promotes Tumor Immunogenicity in Lung Squamous Cell Carcinomas
Authors: DuCote Tanner J. , Song Xiulong , Naughton Kassandra J. , Chen Fan , Plaugher Daniel R. , Childress Avery R. , Gellert Abigail R. , Skaggs Erika M. , Qu Xufeng , Liu Jinze , Liu Jinpeng , Li Fei , Wong Kwok-Kin , Brainson Christine F. 
Published: February 2024

<p>EZH2 inhibition alone and combined with immunotherapy is effective at controlling tumor burden in mouse models of LSCC. <b>A,</b> Representative MRI scans of autochthonous mice from each treatment arm at baseline and after treatment. <b>B,</b> Waterfall plot showing change in tumor volume for each mouse on all treatment arms, ****, <i>P</i> < 0.0001; ***, <i>P</i> < 0.001; **, <i>P</i> < 0.01; *, <i>P</i> < 0.05 by one-way ANOVA with multiple comparisons and Holm-Šídák <i>post hoc</i> test on log<sub>2</sub>-transformed values. <b>C,</b> H&E and HALO nuclear phenotyper images showing the cells within an autochthonous <i>Lkb1/Pten</i> tumor and a syngeneic graft seeded from <i>Lkb1/Pten</i> tumoroids. <b>D,</b> Percentage tumor growth from the syngeneic mouse model during 14 days of indicated treatments. ***, <i>P</i> = 0.0004; ****, <i>P</i> < 0.0001 by one-way ANOVA with multiple comparisons and Holm-Šídák <i>post hoc</i> test, *, <i>P</i> = 0.024 by two-tailed <i>t</i> test on log<sub>2</sub>-transformed values, Mice/tumors n are placebo = 4/8, EPZ6438 = 5/9, anti-PD1 = 6/8, combo = 5/9, mean ± SEM. is plotted. <b>E,</b> 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 <i>n</i> = 7, EZH2 inhibitor <i>n</i> = 7, anti-PD1 <i>n</i> = 8, combo <i>n</i> = 7 with two experimental replicates each, *, <i>P</i> = 0.035; ***, <i>P</i> = 0.0008 by one-way ANOVA with multiple comparisons and Holm-Šídák <i>post hoc</i> test. <b>F,</b> From the same tumor grafts, MFI for HLA-A in the EpCAM+ cells was graphed, mean ± SEM is plotted, placebo <i>n</i> = 7, EZH2 inhibitor <i>n</i> = 7, anti-PD1 <i>n</i> = 8, combo <i>n</i> = 7 with 2 experimental replicates each, **, <i>P</i> = 0.0015 by one-way ANOVA with multiple comparisons and Holm-Šídák <i>post hoc</i> test. <b>G,</b> 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 <i>n</i> = 6, EZH2 inhibitor <i>n</i> = 6, anti-PD1 <i>n</i> = 7, combo <i>n</i> = 7; **, <i>P</i> < 0.006; ***, <i>P</i> = 0.0001; ****, <i>P</i> < 0.0001 by one-way ANOVA with multiple comparisons and Holm-Šídák <i>post hoc</i> test. <b>H,</b> 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 <a href="#SMF5" target="_blank">Supplementary Fig. S5C</a> for representative gates, placebo <i>n</i> = 8, EZH2 inhibitor <i>n</i> = 8, anti-PD1 <i>n</i> = 9, combo <i>n</i> = 7 with two experimental replicates each, *, <i>P</i> = 0.0197; ****, <i>P</i> < 0.0001 by one-way ANOVA with multiple comparisons and Holm-Šídák <i>post hoc</i> test. See also <a href="#SMF5" target="_blank">Supplementary Fig. S5</a>.</p>