Neutrophil Extracellular Traps Correlate with Tumor Necrosis and Size in Human Malignant Melanoma Metastases

Abstract

Neutrophil extracellular traps (NETs) are web-like structures released by neutrophils that kill invading microorganisms. However, NETs also promote tumor growth and impair the functionality of T-cells in cancer. Therefore, this study aimed at characterizing NET distribution within human melanoma metastases (n = 81 of 60 patients) by immunofluorescence staining for neutrophils (CD15) and NETs (H3Cit) in order to identify targets for NET-directed therapies. The results show that 49.3% of the metastases contained neutrophils (n = 40) and 30.8% (n = 25) contained NETs, 68% of them very densely infiltrated. A total of 75% of CD15-positive neutrophils and 96% of NET-containing metastases were necrotic while metastases without neutrophil infiltration were predominantly non-necrotic. A higher amount of NETs correlated significantly with greater tumor size. Consistently, all metastases with a cross-sectional area greater than 2.1 cm2 contained neutrophils. Analysis of metastasis from different sites revealed NETs to be present in skin, lymph node, lung and liver metastases. Taken together, our study was the first to observe NET infiltration in a larger cohort of human melanoma metastases. These results set the stage for further research regarding NET-directed therapies in metastatic melanoma.