凝血因子 XIII-A 转谷氨酰胺酶有助于中性粒细胞胞外诱捕网（NET）介导的纤维蛋白（原）网络形成和交联。
Factor XIII-A Transglutaminase Contributes to Neutrophil Extracellular Trap (NET)-mediated Fibrin(ogen) Network Formation and Crosslinking.
作者信息
Soltani Fatemeh, Welman Mélanie, Samani Sahar Ebrahimi, Pacis Alain, Lordkipanidzé Marie, Kaartinen Mari T
机构信息
Division of Experimental Medicine, Department of Medicine, Faculty of Medicine and Health Sciences, McGill University, Montreal, Canada.
Research Center, Montreal Heart Institute, Montreal, Quebec, Canada.
出版信息
Thromb Haemost. 2025 Jan 20. doi: 10.1055/a-2504-1559.
BACKGROUND
Neutrophil extracellular traps can contribute to thrombosis via stabilization of fibrin network, which is normally conducted by plasma transglutaminase, Factor XIII-A as part of coagulation cascade. The possible presence and activity of FXIII-A in neutrophils or during NETosis are unknown. Here, we investigated potential presence of FXIII-A in neutrophils and participation in NET-fibrin(ogen) interaction in vitro METHODS: Data mining of human and mouse mRNA expression in whole-body scRNA sequence atlases was conducted. mRNA and protein expression was assessed in isolated mouse bone marrow neutrophils. NETosis was induced using 12-phorbol 13-myristate acetate (PMA), and the transglutaminase activity was assessed with 5-(biotinamido)pentylamine incorporation to plasma fibronectin and a fluorescence-fibrin(ogen)-based activity assay using ATTO488-Cadaverine. Externalization of FXIII-A and its interaction with neutrophil extracellular trap (NET) markers, namely, decondensed DNA, CitH3, and MPO, were examined with immunofluorescence microscopy. NET-fibrin(ogen) interaction was investigated with and without serum and/or transglutaminase inhibitor, NC9. Effect of soluble fibrinogen and fibrin(ogen) network on NETosis was also assessed.
RESULTS
Data mining of RNAseq atlases showed expression in adipose tissue, blood, and bone marrow neutrophils. mRNA expression and protein production were confirmed in isolated neutrophils where expression was comparable to that of macrophages and monocytes. FXIII-A was externalized and active as a transglutaminase and colocalized with NET markers during NETosis. FXIII-A transglutaminase activity promoted NET-fibrin(ogen) interaction and entrapment of neutrophils within fibrin(ogen) matrix. Soluble fibrinogen or fibrin(ogen) network did not induce NETosis.
CONCLUSION
This study identifies neutrophils as a source of FXIII-A and suggests its role in stabilizing NET-fibrin(ogen) matrix structures.
背景
中性粒细胞胞外诱捕网可通过稳定纤维蛋白网络促进血栓形成，这一过程通常由血浆转谷氨酰胺酶（凝血因子 XIII-A）作为凝血级联反应的一部分来完成。中性粒细胞中或在中性粒细胞胞外诱捕网形成过程中凝血因子 XIII-A 的可能存在情况及其活性尚不清楚。在此，我们研究了凝血因子 XIII-A 在中性粒细胞中的潜在存在情况以及其在体外参与中性粒细胞胞外诱捕网-纤维蛋白（原）相互作用的情况。
方法
对人和小鼠全身单细胞 RNA 序列图谱中的 mRNA 表达进行数据挖掘。对分离出的小鼠骨髓中性粒细胞的 mRNA 和蛋白质表达进行评估。使用 12-佛波醇 13-肉豆蔻酸酯乙酸酯（PMA）诱导中性粒细胞胞外诱捕网形成，并通过将 5-（生物素酰胺基）戊胺掺入血浆纤连蛋白以及使用 ATTO488-尸胺进行基于荧光纤维蛋白（原）的活性测定来评估转谷氨酰胺酶活性。通过免疫荧光显微镜检查凝血因子 XIII-A 的外化及其与中性粒细胞胞外诱捕网（NET）标志物（即解聚的 DNA、瓜氨酸化组蛋白 H3 和髓过氧化物酶）的相互作用。在有或无血清和/或转谷氨酰胺酶抑制剂 NC9 的情况下研究中性粒细胞胞外诱捕网-纤维蛋白（原）的相互作用。还评估了可溶性纤维蛋白原和纤维蛋白（原）网络对中性粒细胞胞外诱捕网形成的影响。
结果
RNA 测序图谱的数据挖掘显示在脂肪组织、血液和骨髓中性粒细胞中有表达。在分离出的中性粒细胞中证实了 mRNA 表达和蛋白质产生，其表达与巨噬细胞和单核细胞的表达相当。在中性粒细胞胞外诱捕网形成过程中，凝血因子 XIII-A 作为转谷氨酰胺酶外化并具有活性，且与中性粒细胞胞外诱捕网标志物共定位。凝血因子 XIII-A 的转谷氨酰胺酶活性促进了中性粒细胞胞外诱捕网-纤维蛋白（原）的相互作用以及中性粒细胞在纤维蛋白（原）基质中的捕获。可溶性纤维蛋白原或纤维蛋白（原）网络未诱导中性粒细胞胞外诱捕网形成。
结论
本研究确定中性粒细胞是凝血因子 XIII-A 的来源，并表明其在稳定中性粒细胞胞外诱捕网-纤维蛋白（原）基质结构中的作用。
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