Journal: eBioMedicine
Article Title: DNA methylation mediates the immunosuppressive tumour microenvironment in metastatic endometrial clear cell carcinoma
doi: 10.1016/j.ebiom.2025.105954
Figure Lengend Snippet: Epigenetic silencing of the ETS1 regulon compromises anti-tumour immunity . a–g. Analysis of the ECCC sequencing cohort with paired transcriptomic and methylation data (Pn, n = 16; Pm, n = 14). a. Motif enrichment analysis of immune-related hypermethylated DMRs. The y-axis shows the enrichment significance (–log10 (p-value)); the x-axis indicates the motif rank. Significant enrichment was observed for transcription factor binding motifs from the ETS (left) and ZF (right) families. p-Values were determined using a hypergeometric test. b–d. Reduced regulon activity of ETS1 (b, p = 0.002), GATA6 (c, p = 0.038), and PRDM1 (d, p = 0.034) in Pm versus Pn tumours. e. Correlation between ETS1 expression and its target genes. The x-axis represents the Spearman correlation coefficient; the y-axis shows the corresponding p-value. p-Values were calculated by Spearman correlation test. f. Functional enrichment analysis of ETS1 target genes across the GO database. The y-axis shows the enrichment significance (–log10 (q-value)), derived from a hypergeometric test with Benjamini–Hochberg adjustment. The x-axis lists significantly enriched terms. Numbers above bars indicate the count of enriched genes per term. g. Regulatory network of the ETS1 regulon. Transcription factors are represented as ovals and target genes as rectangles; connecting lines indicate regulatory interactions. Oval size corresponds to interaction strength; blue borders denote targets overlapping with downregulated DEGs. h–j. Correlation of ETS1 regulon activity with TME features: TIL density (h, ρ = 0.885, p < 0.001), effector cell signature score (i, ρ = 0.692, p < 0.001), and TLS signature score (j, ρ = 0.726, p < 0.001). p-Values were calculated by Spearman correlation test. k–m, Analyses were performed on the overall ECCC cohort (Pn, n = 27; Pm, n = 24). k. Representative immunohistochemical staining of ETS1. Scale bar = 100 μm. l-m. ETS1 protein expression (H-score) in tumour cells (l, p = 0.248) and immune cells (m, p = 0.002) in Pm versus Pn tumours. All p-values were determined using the Mann–Whitney U test unless otherwise specified. ECCC, endometrial clear cell carcinoma; DMR, differentially methylated region; DMG, differentially methylated gene; Pn, non-metastatic primary tumours; Pm, metastatic primary tumours; DEG, differentially expressed gene; TME, tumour microenvironment; TIL, tumour-infiltrating lymphocyte; TLS, tertiary lymphoid structure.
Article Snippet: Endogenous peroxidase activity was blocked with 3% hydrogen peroxide for 10 min, followed by incubation with primary ETS1 antibody (Cat#66598-1-Ig, RRID: AB_2881958 , Proteintech, China) overnight at 4 °C.
Techniques: Sequencing, Methylation, Binding Assay, Activity Assay, Expressing, Functional Assay, Derivative Assay, Immunohistochemical staining, Staining, MANN-WHITNEY