A comprehensive plasma-based approach to thromboinflammation in bladder cancer: integrating lipidomics, thrombin generation, and NETosis biomarkers.

BACKGROUND

Lipid metabolism reprogramming is a hallmark of cancer, yet its systemic manifestations and functional implications in bladder cancer (BC) remain poorly understood. This study aimed to identify plasma lipidomic profiles with diagnostic, staging, and prognostic value in BC, and to explore their associations with thromboinflammatory mechanisms, particularly NETosis and thrombin generation.

METHODS

Plasma lipidomics was characterized by ultra-high-performance liquid chromatography high-resolution mass spectrometry from 87 BC patients (non-muscle-invasive BC, NMIBC, and muscle-invasive BC, MIBC) and 30 controls. NETosis markers, active DNaseI concentration, C-reactive protein, fibrinogen, IL-6, prothrombin F1+2, thrombin-antithrombin (TAT) complexes, and thrombin generation tests (TGT) were assessed in parallel. Statistical analyses in R included PCA, ROC curve analysis, elastic net penalized logistic regression, Cox regression, natural cubic spline modeling for survival analyses, and pathway enrichment analyses.

RESULTS

BC patients exhibited increased total lipid load and distinct lipidomic signatures. Phosphatidylcholine (PC) levels decreased with disease severity and were independently associated with poor overall survival (up to 85% reduction in MIBC). Multivariable elastic net models integrating lipidomic and clinical variables achieved high diagnostic accuracy for discriminating BC patients from controls (AUC = 0.943) and for distinguishing MIBC from NMIBC (AUC = 0.860). BC patients displayed increased NETosis, inflammation, and thrombin formation, while decreased active DNaseI. Potentially diagnostic lipids correlated with NETosis, inflammatory markers and TAT, and inversely with active DNaseI and TGT parameters. Given the cross-sectional and observational design, these findings should be interpreted as hypothesis-generating and do not allow causal inference.

CONCLUSIONS

Plasma lipidomic profiling identifies biomarkers with diagnostic and prognostic potential in BC and reveals associations with NETosis, active DNaseI dysfunction, and thrombin generation. While causality cannot be inferred from this exploratory study, these findings support lipidomics as a minimally invasive tool for BC stratification and as a window into cancer-associated immunothrombosis, providing a rationale for future longitudinal and mechanistic studies targeting lipid-neutrophil interactions.