Fig. 5: IRF2 and POU2F1 repression by mutated-β-catenin is a major tumor cell-intrinsic mechanism of immune exclusion in CTNNB1-mutated HCC.
From: Precision targeting of β-catenin induces tumor reprogramming and immunity in hepatocellular cancers
a Schematic of pipeline comparing whole transcriptomes of β-catenin-mutated HCC to β-catenin knockout livers. b Dot plot highlighting Irf2 and Pou2f1 target genes within the Zone 3 CTNNB1 WT and MUT (GS+) cell population between LNP-CTRL and LNP-CTNNB1 from Fig. 3a, b. c Heatmap of z-scored expression values of IRF2/POU2F1 target genes in TCGA-LIHC patients (n = 374) and adjacent normal (n = 50). Data stratified by CTNNB1- (n = 98), AXIN1- (n = 18), and APC-mutated patients (n = 3). d Boxplots of normalized expression of IRF2, POU2F1, and IRF2/POU2F1 target genes stratified by adjacent normal (n = 50), Wnt/β-catenin-mutant (n = 119), and -wild-type (n = 255). e Schematic of β-catenin-hMet (β-M) animals co-injected with pT3 or IRF2 at time of hydrodynamic tail vein injection (HDTVi) and sacrificed at 7.5-weeks post-HDTVi. f Representative gross liver images from β-M-pT3 and β-M-IRF2 animals. Scale bar indicates 1 centimeter (cm). g, h Liver weights (p = 0.0008) and liver weight/body weight (LW/BW) (p = 0.0003) comparing β-M-pT3 (n = 7) and β-M-IRF2 (n = 12) animals at 7.5-week timepoint. i Schematic of β-catenin-Nrf2 (β-N) animals co-injected with pT3 or POU2F1 at time of HDTVi and sacrificed at 10.7-weeks post-HDTVi. j Representative gross liver images from β-N-pT3 and β-N-POU2F1 animals. k, l Liver weights (p = 0.0013) and LW/BW (p = 0.0005) comparing β-N-pT3 (n = 4) and β-N-POU2F1 (n = 4) animals at 10.7-week timepoint. m Representative IHC images for CD4, CD8, and CD20 comparing β-N-pT3 and β-N-POU2F1 animals at 10.7-week timepoint. n GO pathway gene set enrichment analysis comparing β-M-POU2F1 to β-M-pT3. Created in BioRender. Lehrich (2025) https://BioRender.com/25qvbwj. Lehrich (2025) https://BioRender.com/qqvvb4x. Lehrich (2025) https://BioRender.com/ecs8omd. For (g, h), (k, l), data presented as mean values ± standard deviation (SD) and P-values calculated by unpaired two-tailed Student’s t-test. For (d), the center line shows the median, the box limits show the interquartile range (IQR; the range between the 25th and 75th percentile) and the whiskers show 1.5× IQR. For (d), One-way ANOVA with Tukey-HSD post-hoc adjusted p-values comparing Wnt/β-catenin-mutant vs wild-type are: p = 0.80, p = 0.54, and p = 0.009 for IRF2, POU2F1, and IRF2/POU2F1 target gene expression, repsectively. For (n), NES was normalized from ES, which was calculated by Kolmogorov–Smirnov-like statistic and the adjusted p-value was calculated using the one-tailed empirical permutation test procedure. Source data are provided as a Source Data File. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
