Natural killer cell-related gene signature predicts immune cell infiltration and improved survival in bladder cancer

Natural killer (NK) cell-based therapies represent a promising immunotherapeutic approach for cancer treatment. This study aims to identify and evaluate the prognostic significance of NK cell-related genes in bladder cancer (BC).

Using RNA-seq data from The Cancer Genome Atlas (TCGA) BLCA cohort, an NK cell-related gene signature was constructed and validated via multivariate Cox regression. Functional enrichment analyses (GO and KEGG) were conducted to explore associated biological processes. Immune infiltration states were assessed using ESTIMATE and CIBERSORT algorithms. Immunohistochemistry (IHC) and multiplex immunofluorescence (mIF) were performed to validate key findings at the protein level.

An eight-gene prognostic signature based on NK cell-related genes was established, enabling stratification of BC patients into high- and low-risk groups. Patients in the high-risk group exhibited significantly worse overall survival (OS) and progression-free survival (PFS). Multivariate analysis confirmed the signature's independence from other clinical variables. The signature was also correlated with distinct immune infiltration patterns, including CD8⁺ T cells, CD4⁺ activated memory T cells, follicular helper T cells, regulatory T cells, activated NK cells, activated dendritic cells, and M0/M2 macrophages. Notably, the key signature gene RAC3 was found to be highly expressed in a subset of tumors. This elevated RAC3 expression was associated with enhanced proliferation (increased Ki67) and reduced infiltration of CD8⁺ T cells and CD56⁺ NK cells, indicating an immunosuppressive microenvironment. Consistently, the TIDE score was significantly lower in the high-risk group, suggesting enhanced potential responsiveness to immunotherapy.

The NK cell-related gene signature serves as a robust prognostic biomarker and predictor of immunotherapeutic efficacy in BC. These findings provide new insights into BC biology and facilitate personalized immunotherapy strategies.