Elevated neutrophil extracellular traps as potential diagnostic biomarkers in pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most aggressive malignancies with limited diagnostic and prognostic markers. Neutrophil extracellular traps (NETs) have recently been implicated in cancer progression, but their clinical relevance in PDAC remains underexplored. In this preliminary study, we investigated NET levels in both peripheral blood and tumor tissues from PDAC patients (n = 30) and explored their associations with clinicopathological features. We quantified NETs using a multiparametric approach that included immunohistochemistry, immunofluorescence, qRT-PCR, ELISA, and flow cytometry. Plasma NET-associated DNA was measured using the Quant-iT PicoGreen assay. Receiver operating characteristic (ROC) analysis was performed to assess the diagnostic potential of NETs. We found NET levels were significantly elevated in both tumor tissue and the circulation of PDAC patients compared with healthy controls. The levels of NETs were strikingly higher in patients with advanced tumor stages and grades. Patients' NETs showed a typical morphology with enlarged nuclei, thread-like DNA fibres, and MPO-positive granules, which was confirmed by immunofluorescence analysis. The ROC analysis demonstrated that NETs displayed promising diagnostic performance (AUC = 0.852) compared to widely used conventional markers CEA and CA19-9 within the study cohort. In conclusion, our findings provide preliminary evidence that increased NETosis is associated with PDAC progression and highlight its potential as a diagnostic biomarker. Larger, independent studies are needed to validate these observations and to determine the clinical relevance of NET-associated markers in PDAC.