Figure 4. Kmt2d-mutant tumors are associated with increased innate and adaptive immune infiltration.

(A) Schematic of experimental design for generating the syngeneic LLC dual-tumor model. LLC-Vector and LLC-sgKmt2d cells were transplanted into the left and right flanks of C57BL/6J mice, respectively, followed by treatment with PBS or anti-PD1. Tumor-infiltrating immune cells were analyzed by FACS analysis.

(B) Abundance of intratumoral CD45⁺ pan-immune cells, CD4⁺ T cells, CD8⁺ T cells, and IFN-γ⁺ CD8⁺ T cells in LLC-Vector or LLC-sgKmt2d tumors, treated with PBS (n ≥ 5) or aPD1 (n ≥ 5). Mann-Whitney test: sgKmt2d + aPD1 vs. Vector + aPD1: CD45⁺ cells, P = 0.052; CD4⁺ T cells, P = 0.0029; CD8⁺ T cells, P = 0.0003; IFN-γ⁺ CD8⁺ T cells, P = 0.0079.

(C) The intratumoral abundance of monocytes, neutrophils, dendritic cells, macrophages, TAM1 (tumor-associated macrophage 1) and TAM2 in LLC-Vector or LLC-sgKmt2d tumors, treated with PBS (n = 5) or aPD1 (n = 5). Mann-Whitney test, sgKmt2d + aPD1 vs Vector + aPD1: monocytes, P = 0.4206; neutrophils, P = 0.3095; dendritic cells, P = 0.0159; macrophages, P = 0.0079; TAM1, P = 0.0317; TAM2, P = 0.0159.

(D) PD1-expression in different immune populations present within LLC-Vector or LLC-sgKmt2d tumors, treated with PBS (n = 5) or aPD1 (n = 5). Mann-Whitney test, sgKmt2d + aPD1 vs Vector + aPD1: PD1⁺CD4⁺ T cells, P = 0.2222; PD1⁺CD8⁺ T cells, P = 0.2222; PD1⁺ macrophages, P = 0.4206; PD1⁺monocytes, P = 0.6905.

(E) Volcano plot of the Spearman correlation between KMT2D mRNA expression and macrophage abundance in 33 cancer types. Red dots indicate cancer types in which KMT2D is significantly negatively correlated with macrophage abundance (adjusted P < 0.05).

(F) Scatter plots comparing KMT2D expression and macrophage abundance in TCGA BLCA, BRCA, LUSC, and LIHC cohorts.

Error bars: All data points in this figure are presented as mean ± s.e.m. Asterisks: * P < 0.05, ** P < 0.01, *** P < 0.001.

See also: Supplementary Fig. S4, S5, S6