PMID- 35095884 OWN - NLM STAT- MEDLINE DCOM- 20220223 LR - 20220223 IS - 1664-3224 (Electronic) IS - 1664-3224 (Linking) VI - 12 DP - 2021 TI - Increased Circulating Cell-Free DNA in Eosinophilic Granulomatosis With Polyangiitis: Implications for Eosinophil Extracellular Traps and Immunothrombosis. PG - 801897 LID - 10.3389/fimmu.2021.801897 [doi] LID - 801897 AB - BACKGROUND: Endogenous DNA derived from nuclei or mitochondria is released into the blood circulation as cell-free DNA (cfDNA) following cell damage or death. cfDNA is associated with various pathological conditions; however, its clinical significance in antineutrophil cytoplasmic antibody-associated vasculitis (AAV) remains unclear. This study aimed to evaluate the clinical significance of cfDNA in AAV. METHODS: We enrolled 35 patients with AAV, including 10 with eosinophilic granulomatosis with polyangiitis (EGPA), 13 with microscopic polyangiitis, and 12 with granulomatosis with polyangiitis. Serum cf-nuclear DNA (cf-nDNA) and cf-mitochondrial DNA (cf-mtDNA) levels were measured by quantitative polymerase chain reaction before and after the initiation of immunosuppressive therapy. Tissue samples from EGPA patients were examined by immunofluorescence and transmission electron microscopy. The structure of eosinophil extracellular traps (EETs) and neutrophil extracellular traps (NETs) and stability against DNase were assessed in vitro. Platelet adhesion of EETs were also assessed. RESULTS: Serum cf-nDNA and cf-mtDNA levels were significantly higher in AAV than in healthy controls, with the highest levels in EGPA; however, serum DNase activities were comparable among all groups. cf-nDNA and cf-mtDNA decreased after treatment and were associated with disease activity only in EGPA. Blood eosinophil count and plasma D-dimer levels were significantly correlated with cf-nDNA in EGPA and cf-mtDNA. EGPA tissue samples showed lytic eosinophils and EETs in small-vessel thrombi. The structure of EETs showed bolder net-like chromatin threads in vitro and EETs showed greater stability against DNase than NETs. EETs provided a scaffold for platelet adhesion. CONCLUSION: cfDNA was increased in EGPA, associated with disease activity. The presence of DNase-resistant EETs in small-vessel thrombi might contribute to higher concentration of cfDNA and the occurrence of immunothrombosis in EGPA. CI - Copyright (c) 2022 Hashimoto, Ueki, Kamide, Miyabe, Fukuchi, Yokoyama, Furukawa, Azuma, Oka, Takeuchi, Kanno, Ishida-Yamamoto, Taniguchi, Hashiramoto and Matsui. FAU - Hashimoto, Teppei AU - Hashimoto T AD - Division of Diabetes, Endocrinology and Clinical Immunology, Department of Internal Medicine, Hyogo College of Medicine, Nishinomiya, Japan. FAU - Ueki, Shigeharu AU - Ueki S AD - Department of General Internal Medicine and Clinical Laboratory Medicine, Akita University Graduate School of Medicine, Akita, Japan. FAU - Kamide, Yosuke AU - Kamide Y AD - National Hospital Organization Sagamihara National Hospital, Clinical Research Center, Sagamihara, Japan. FAU - Miyabe, Yui AU - Miyabe Y AD - Department of General Internal Medicine and Clinical Laboratory Medicine, Akita University Graduate School of Medicine, Akita, Japan. FAU - Fukuchi, Mineyo AU - Fukuchi M AD - Department of General Internal Medicine and Clinical Laboratory Medicine, Akita University Graduate School of Medicine, Akita, Japan. FAU - Yokoyama, Yuichi AU - Yokoyama Y AD - Pathology and Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, United States. FAU - Furukawa, Tetsuya AU - Furukawa T AD - Division of Diabetes, Endocrinology and Clinical Immunology, Department of Internal Medicine, Hyogo College of Medicine, Nishinomiya, Japan. FAU - Azuma, Naoto AU - Azuma N AD - Division of Diabetes, Endocrinology and Clinical Immunology, Department of Internal Medicine, Hyogo College of Medicine, Nishinomiya, Japan. FAU - Oka, Nobuyuki AU - Oka N AD - Department of Neurology, Kyoto Konoe Rehabilitation Hospital, Kyoto, Japan. FAU - Takeuchi, Hiroki AU - Takeuchi H AD - Department of Neurology, National Hospital Organization Minami Kyoto Hospital, Kyoto, Japan. FAU - Kanno, Kyoko AU - Kanno K AD - Department of Dermatology, Asahikawa Medical University, Asahikawa, Japan. FAU - Ishida-Yamamoto, Akemi AU - Ishida-Yamamoto A AD - Department of Dermatology, Asahikawa Medical University, Asahikawa, Japan. FAU - Taniguchi, Masami AU - Taniguchi M AD - National Hospital Organization Sagamihara National Hospital, Clinical Research Center, Sagamihara, Japan. FAU - Hashiramoto, Akira AU - Hashiramoto A AD - Department of Biophysics, Kobe University Graduate School of Health Sciences, Kobe, Japan. FAU - Matsui, Kiyoshi AU - Matsui K AD - Division of Diabetes, Endocrinology and Clinical Immunology, Department of Internal Medicine, Hyogo College of Medicine, Nishinomiya, Japan. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20220112 PL - Switzerland TA - Front Immunol JT - Frontiers in immunology JID - 101560960 RN - 0 (Antibodies, Antineutrophil Cytoplasmic) RN - 0 (Biomarkers) RN - 0 (Cell-Free Nucleic Acids) RN - 0 (Fibrin Fibrinogen Degradation Products) RN - 0 (Immunosuppressive Agents) RN - 0 (fibrin fragment D) SB - IM MH - Aged MH - Antibodies, Antineutrophil Cytoplasmic/blood/immunology MH - Biomarkers MH - Blood Platelets/immunology/metabolism/pathology/ultrastructure MH - *Cell-Free Nucleic Acids MH - Diagnosis, Differential MH - Disease Susceptibility/immunology MH - Eosinophils/*immunology/*metabolism/*pathology MH - Extracellular Traps/immunology/metabolism MH - Female MH - Fibrin Fibrinogen Degradation Products MH - Granulomatosis with Polyangiitis/diagnosis/*etiology/*metabolism/therapy MH - Humans MH - Immunosuppressive Agents/therapeutic use MH - Kidney Function Tests MH - Leukocyte Count MH - Liquid Biopsy/methods MH - Male MH - Microscopic Polyangiitis/diagnosis MH - Middle Aged MH - Platelet Aggregation MH - *Thromboinflammation/complications/diagnosis/etiology/immunology PMC - PMC8790570 OTO - NOTNLM OT - ANCA-associated vasculitis OT - cell-free DNA OT - eosinophil extracellular traps (EETs) OT - eosinophilic granulomatosis with polyangiitis OT - immunothrombosis OT - thrombosis COIS- The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2022/02/01 06:00 MHDA- 2022/02/24 06:00 PMCR- 2021/01/01 CRDT- 2022/01/31 05:56 PHST- 2021/10/26 00:00 [received] PHST- 2021/12/20 00:00 [accepted] PHST- 2022/01/31 05:56 [entrez] PHST- 2022/02/01 06:00 [pubmed] PHST- 2022/02/24 06:00 [medline] PHST- 2021/01/01 00:00 [pmc-release] AID - 10.3389/fimmu.2021.801897 [doi] PST - epublish SO - Front Immunol. 2022 Jan 12;12:801897. doi: 10.3389/fimmu.2021.801897. eCollection 2021.