Fig. 3: Caspase-1-mediated IL1β processing within TNBC cells contributes to a functionally and spatially distinct TIME.
From: Caspase-1-dependent spatiality in triple-negative breast cancer and response to immunotherapy
a Schematic of IL1β-P2A where P2A25 is inserted at the caspase-1 cleavage site within the IL1β gene, allowing caspase-independent processing of IL1β via endogenous ribosomal skipping. Created with BioRender.com, released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license. Growth curves, tumor weights, flow cytometric quantification (F4/80 and CD11b double positive cells as a proportion of CD45 positive immune cells) and IHC quantification of F4/80 positive cells in wild type (n = 5), ERα-overexpressing (n = 6) and ERα-overexpressing P2A rescue KBP (n = 8) allografts, tumor weight *: p = 0.024, F4/80+ CD11b+ in CD45+ *: p = 0.0137, **: p = 0.0042, ***: p = 0.00027 (b), in KBP shETS1 allografts with and without P2A rescue in mice fed regular (-dox) or doxycycline-containing (+dox) diet at tumor onset (n = 5 mice/group), tumor weight *: p = 0.0446, F4/80+ CD11b+ in CD45+ *: p = 0.0143, F4/80 in total cells *: p = 0.0197, ***: p = 0.0007, ****: p < 0.0001 (c), in wild type, caspase-1 KO and caspase-1 KO P2A KBP allografts (n = 7 mice/group), tumor volume *: p = 0.0326, F4/80 in total cells *: p = 0.0316, tumor volume ***: p = 0.0001; tumor weight ***: p = 0.0001; F4/80+ CD11b+ in CD45+ ***: p = 0.0007 (d), and in mock-treated wild type KBP, VX-765-treated KBP and VX-765-treated P2A rescue KBP allografts (n = 6 mice/group), tumor weight **: p = 0.0039; F4/80+ CD11b+ in CD45+ **: p = 0.004, ***: p = 0.0002, ****: p < 0.0001 (e). Representative IHC images (f) and quantification (g) of the location (expressed as percentage of total cells in tumor core/tumor periphery) of CD3+ and CD8+ cells in wild type, caspase-1 KO and VX-765-treated KBP allografts (n = 3 mice/group), CD3+ wild type vs caspase-1 KO: ** p = 0.0067; wild type vs wild type + VX-765: **: p = 0.0011; CD8+ wild type vs caspase-1 KO: **: p = 0.0053; wild type vs wild type + VX-765: **: p = 0.0027; Mean with SEM, two-way ANOVA with Tukey’s multiple comparisons test for growth curves (b–e), one-way ANOVA with Dunnett’s multiple comparisons test for tumor weights and immune cell quantification (b, d, e, g), one-way ANOVA with Tukey’s multiple comparisons test for tumor weights and macrophage quantification (c). ns = not significant, scalebar corresponds to 50μm.
