多形核中性粒细胞（PMNs）是炎症反应中的主要效应细胞，在甲状腺癌（TC）中扮演多种角色。PMNs包含并释放大量介质，包括颗粒酶（例如髓过氧化物酶（MPO），蛋白质-3（PTX3）和基质金属蛋白酶-9（MMP-9）），以及中性粒细胞外陷阱（NETs）。本研究旨在评估NETs和中性粒细胞来源的介质作为可能的TC患者生物标志物。招募了20例分化型甲状腺癌（DTC）患者，26例去分化型甲状腺癌（De-DTC）患者，26例多结节性甲状腺肿（MNG）患者和22例健康对照组（HCs）。通过酶联免疫吸附法（ELISA）测定了游离DNA（dsDNA），核小体，精氨酸化组蛋白H3（CitH3）和MPO-DNA复合物的血清浓度作为NET生物标志物。通过ELISA测量了MPO，PTX3，MMP-9，CXCL8和粒细胞单核细胞集落刺激因子（GM-CSF）等中性粒细胞相关介质的血清水平。与HCs相比，De-DTC患者的四种NET生物标志物的血清水平均增加。CitH3血清水平在De-DTC和DTC患者中有选择性增加，与HCs和MNG患者相比。MPO-DNA复合物和核小体仅在De-DTC患者中有选择性增加，与HCs和MNG患者相比。此外，MPO-DNA复合物在De-DTC患者中相对于DTC患者也有选择性增加。与HCs相比，MPO循环水平在De-DTC患者亚组中有选择性增加。与HCs相比，DTC和De-DTC患者的PTX3，MMP-9和GM-CSF循环水平均增加。核小体与dsDNA，CitH3，MPO和CXCL8呈正相关。MPO-DNA复合物与dsDNA，CitH3，CXCL8，MPO和核小体水平呈正相关。此外，四种NET生物标志物中的三种（即dsDNA，核小体和MPO-DNA复合物）在老年患者中相对于年轻患者增加，在诊断时有转移性疾病的患者中相对于无转移的患者增加。男性的核小体比女性高。MPO-DNA复合物，核小体，在某种程度上CitH3水平似乎与进展性甲状腺癌的恶性程度相关。此外，与MNG和HCs相比，TC患者的PMN相关介质（MPO，PTX3，GM-CSF）的血清浓度增加。Copyright © 2023 Modestino，Cristinziano，Poto，Ventrici，Trocchia，Ferrari，Fallahi，Paparo，Marone，Antonelli，Varricchi和Galdiero。

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.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.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.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.Copyright © 2023 Modestino, Cristinziano, Poto, Ventrici, Trocchia, Ferrari, Fallahi, Paparo, Marone, Antonelli, Varricchi and Galdiero.