Abstract
Background: As a member of the tumor necrosis factor (TNF) family, TNF superfamily 4 (TNFSF4) plays a crucial role in various immune-related processes. However, its biological function in pan-cancer remains largely unexplored. Methods: RNA-sequencing (RNA-seq) data and corresponding clinical variables were obtained from the Cancer Genome Atlas (TCGA). Immunotherapy cohorts were retrieved from Gene Expression Omnibus (GEO). Tumor Immune Estimation Resource was used to evaluate tumor-infiltrating immune cell levels. The Tumor Immune Single-cell Hub (TISCH) 2 database was used to examine TNFSF4 expression across various tumor cell subsets. Gene set enrichment analysis (GSEA) was performed to investigate TNFSF4-associated signaling pathways. Results: Bioinformatic analyses of TNFSF4 across TCGA cancers revealed that TNFSF4 expression was highly increased in cancers compared to normal tissues. Further immunohistochemistry staining of multiple tumor samples validated this finding. Univariate Cox regression and survival analyses identified TNFSF4 as a risk factor in most cancers. Furthermore, TNFSF4 expression increased with tumor stage progression in several cancers. In the two immunotherapy cohorts from the GEO database, the proportion of partial response/complete response patients was higher in the TNFSF4 low-expression group. Single-cell RNA-seq data from the TISCH database indicated that TNFSF4 expression was predominantly observed in proliferative and exhausted T cells. Correlation analysis demonstrated a positive association between TNFSF4 and immunomodulatory genes. Ultimately, GSEA revealed that TNFSF4 was related to immune response and epithelial-mesenchymal transition pathways. Conclusions: This multi-omics analysis highlights the role of TNFSF4 in tumor progression and immune modulation. High TNFSF4 expression correlates with poor survival and may affect immunotherapy efficacy, suggesting its potential prognostic biomarker and therapeutic target.