Neutrophil extracellular traps and neutrophil-related mediators in human thyroid cancer

Luca Modestino; Leonardo Cristinziano; Remo Poto; Annagioia Ventrici; Marialuisa Trocchia; Silvia Martina Ferrari; Poupak Fallahi; Sabrina Rosaria Paparo; Gianni Marone; Alessandro Antonelli; Gilda Varricchi; Maria Rosaria Galdiero

doi:10.3389/fimmu.2023.1167404

Neutrophil extracellular traps and neutrophil-related mediators in human thyroid cancer

Front Immunol. 2023 Aug 29:14:1167404. doi: 10.3389/fimmu.2023.1167404. eCollection 2023.

Authors

Luca Modestino^{1

2

3}, Leonardo Cristinziano^{1

2

3}, Remo Poto^{1

2

3}, Annagioia Ventrici^{1

2

3}, Marialuisa Trocchia^{1

2

3}, Silvia Martina Ferrari⁴, Poupak Fallahi⁵, Sabrina Rosaria Paparo⁶, Gianni Marone^{1

2

3

7}, Alessandro Antonelli⁶, Gilda Varricchi^{1

2

3

7}, Maria Rosaria Galdiero^{1

2

3}

Affiliations

¹ Department of Translational Medical Sciences (DiSMeT), University of Naples Federico II, Naples, Italy.
² Center for Basic and Clinical Immunology Research (CISI), University of Naples Federico II, Naples, Italy.
³ World Allergy Organization Center of Excellence, University of Naples Federico II, Naples, Italy.
⁴ Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
⁵ Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Pisa, Italy.
⁶ Department of Surgical, Medical and Molecular Pathology and Critical Area, University of Pisa, Pisa, Italy.
⁷ Institute of Experimental Endocrinology and Oncology (IEOS), National Research Council (CNR), Naples, Italy.

Abstract

Background: Polymorphonuclear neutrophils (PMNs) are the main effector cells in inflammatory responses and play multiple roles in thyroid cancer (TC). PMNs contain and release a plethora of mediators, including granular enzymes [e.g., myeloperoxidase (MPO), pentraxin-3 (PTX3) and matrix metalloproteinase-9 (MMP-9)], and neutrophil extracellular traps (NETs). The aim of this study was to evaluate NETs and neutrophil-derived mediators as possible biomarkers in TC patients.

Methods: 20 patients with differentiated thyroid cancer (DTC), 26 patients with dedifferentiated thyroid cancer (De-DTC), 26 patients with multinodular goiter (MNG) and 22 healthy controls (HCs) were recruited. Serum concentrations of free DNA (dsDNA), nucleosomes, citrullinated histone H3 (CitH3) and MPO-DNA complexes were evaluated as NET biomarkers. Neutrophil-related mediators such as MPO, PTX3, MMP-9, CXCL8, and granulocyte-monocyte colony-stimulating factor (GM-CSF) were measured by ELISA.

Results: Serum levels of all four NET biomarkers were increased in DeDTC patients compared to HCs. CitH3 serum levels were selectively increased in both DeDTC and DTC patients compared to HCs and MNG patients. MPO-DNA complexes and nucleosomes were selectively increased only in DeDTC patients compared to HCs and MNG patients. Moreover, MPO-DNA complexes were selectively increased in DeDTC patients compared to DTC patients also. MPO circulating levels were selectively increased in the DeDTC patient subgroup compared to HCs. Circulating levels of PTX3, MMP-9 and GM-CSF were increased in DTC and DeDTC patients compared to HCs. Nucleosomes positively correlated with dsDNA, CitH3, MPO and CXCL8. MPO-DNA complexes positively correlated with dsDNA, CitH3, CXCL8, MPO and nucleosome levels. Moreover, three out of the four NET biomarkers (i.e., dsDNA, nucleosomes and MPO-DNA complexes) were increased in elderly patients compared to young patients and in patients with metastatic disease at diagnosis compared to non metastatic patients. Nucleosomes were higher in males compared to females.

Conclusion: MPO-DNA complexes, nucleosomes and, to some extent, CitH3 levels seem to correlate with malignancy and severity of progressive TC. Moreover, serum concentrations of PMN-related mediators (MPO, PTX3, GM-CSF) were increased in TCs compared to MNG and HCs.

Keywords: NETs biomarkers; dedifferentiated thyroid cancer; neutrophil extracellular traps; neutrophils; thyroid cancer.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenocarcinoma*
Aged
Extracellular Traps*
Female
Granulocyte-Macrophage Colony-Stimulating Factor
Histones
Humans
Male
Matrix Metalloproteinase 9
Neutrophils
Nucleosomes
Thyroid Neoplasms*

Substances

Matrix Metalloproteinase 9
Nucleosomes
Granulocyte-Macrophage Colony-Stimulating Factor
Histones

Grants and funding

This work was supported in part by grants from the CISILab Project (University of Naples Federico II), TIMING Project and Campania Bioscience (Regione Campania) and by MIUR-PRIN 2017M8YMR8_005 and AIRC under MFAG 2020 (grant number 25123) to MRG.