Figure 8 from OPG-Producing B Cells and RANKL-Expressing T Cells Define Immune Signatures Predictive of Bone Metastases in Breast Cancer

Divergent immune signatures defined by RANKL and OPG expression are associated with differential metastatic risk in BRCA-Basal breast tumors. A, Spearman correlation analysis between TNFSF11 (RANKL) gene expression and CD4+ T cell subpopulations in BRCA-Basal tumors (n = 191, TCGA dataset). Significant positive correlations were observed with total CD4+ T cells (EPIC, TIMER), naive and memory subsets (CIBERSORT, XCELL), and Tregs (QUANTISEQ). An inverse correlation with tumor purity suggests an immune (rather than tumor) source of RANKL expression. B,Z-score risk analysis identified high infiltration of CD4+ central memory T cells as significantly associated with increased clinical risk in the BRCA-Basal subtype (Z = 2.253; P < 0.05). C, Spearman correlations between TNFRSF11B (OPG) expression and B cell subpopulations in BRCA-Basal tumors. Significant positive associations were detected with total B cells (QUANTISEQ), naive B cells, and class-switched memory B cells (XCELL), alongside a negative correlation with tumor purity, reinforcing an immune origin for OPG expression. D,Z-score risk analysis revealed that high plasma B cell infiltration was significantly associated with reduced clinical risk (Z = −2.078; P < 0.05), implicating a protective immune profile related to OPG expression. Data were obtained from the TCGA cohort using TIMER 2.0 and analyzed through multiple deconvolution algorithms (EPIC, TIMER, CIBERSORT, XCELL, QUANTISEQ). A and D, Correlation analyses between RANKL (Tnfsf11) or OPG (Tnfrsf11b) expression and immune cell infiltration across the BRCA-Basal TCGA cohort (n = 191) were performed using the TIMER, EPIC, CIBERSORT, XCELL, and QUANTISEQ deconvolution platforms. Statistical significance was determined by Spearman’s rank correlation test. B and C, Clinical risk associations for each immune subset were computed using Z-score analysis (two-tailed). Significance thresholds: P < 0.05.