Elucidation of DNase resistance acquisition mechanism of neutrophil extracellular traps in ANCA-associated vasculitis

• Project/Area Number: 19K16575
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 49020:Human pathology-related
• Research Institution: Hokkaido University
• Principal Investigator: Masuda Sakiko 北海道大学, 保健科学研究院, 講師 (10763617)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2021: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2020: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2019: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: ANCA関連血管炎 / 好中球細胞外トラップ / NETs / DNase I / プロピルチオウラシル / DNase抵抗性NETs / 唾液

Outline of Research at the Start

抗好中球細胞質抗体（ANCA）関連血管炎の病因病態形成において、好中球細胞外トラップ（NETs）が関与しており、生体内の主要なNETs分解因子であるDNase Iに抵抗性を示すNETsが形成されている。本研究では、DNase I抵抗性NETsの形成機序を明らかにすることを目的とする。DNase I感受性NETsとDNase I抵抗性NETsのタンパク質をプロテオーム解析により比較し、NETsにDNase I抵抗性を付与するタンパクを明らかにし、DNase I感受性NETsへと導く薬剤候補を明らかにする。

Outline of Final Research Achievements

The proteins of DNase I-sensitive NETs or DNase I-resistant NETs were compared by proteomics analysis, and narrowed down 18 protein candidates that are considered to confer DNase I resistance to NETs. When candidate proteins were added during NET induction, the formation of normal NET was suppressed by the five proteins, and the formation of abnormal NET resistant to DNase I became the main component. Addition of an inhibitory antibody to one of these proteins during NETs induction suppressed the formation of normal and abnormal NETs.

Academic Significance and Societal Importance of the Research Achievements

ANCA関連血管炎では分解因子DNase Iに抵抗性を持つNETsが産生され、その結果自己抗体の産生や組織障害が誘導される。本研究では、NETsにDNase I抵抗性を付与する可能性のあるタンパクを絞り込んだ。そのうちの１つのタンパクに対する阻害抗体は異常なDNase I抵抗性NETsの産生を抑制したことから、本研究の成果はDNase I抵抗性NETsが関連する疾患の治療に応用できる可能性がある。

Research Products

• [Journal Article] Involvement of neutrophil extracellular traps in the pathogenesis of glomerulonephritis in a case of systemic lupus erythematosus and antineutrophil cytoplasmic antibody-associated vasculitis overlap syndrome (2022)
  • Author(s): Senda Arisa、Sasai Ryutaro、Kato Kurumi、Nishibata Yuka、Masuda Sakiko、Ishizu Akihiro、Takahara Noriko
  • Journal Title: CEN Case Reports Volume: - Issue: 3 Pages: 339-346
  • DOI: 10.1007/s13730-021-00682-y
  • Peer Reviewed
• [Journal Article] Possible implication of intermolecular epitope spreading in the production of anti-glomerular basement membrane antibody in anti-neutrophil cytoplasmic antibody-associated vasculitis (2022)
  • Author(s): Yuka Nishibata, Mayu Nonokawa, Yuto Tamura, Rio Higashi, Ku Suzuki, Hideyuki Hayashi, Sakiko Masuda, Daigo Nakazawa, Satoshi Tanaka, Utano Tomaru, Akihiro Ishizu
  • Journal Title: Clinical and Experimental Rheumatology Volume: - Pages: 691-704
  • DOI: 10.55563/clinexprheumatol/6oq9du
  • Peer Reviewed / Open Access
• [Journal Article] Phorbol 12-myristate 13-acetate stimulation under hypoxia induces nuclear swelling with DNA outflow but not extracellular trap formation of neutrophils (2022)
  • Author(s): Masuda Sakiko、Kato Kurumi、Ishibashi Misato、Nishibata Yuka、Sugimoto Ayako、Nakazawa Daigo、Tanaka Satoshi、Tomaru Utano、Tsujino Ichizo、Ishizu Akihiro
  • Journal Title: Experimental and Molecular Pathology Volume: 125 Pages: 104754-104754
  • DOI: 10.1016/j.yexmp.2022.104754
  • Peer Reviewed / Open Access
• [Journal Article] Spontaneously regressed granulomatosis with polyangiitis: A case report (2021)
  • Author(s): Ota Hiroki、Sato Chisa、Igarashi Akira、Inoue Sumito、Masuda Sakiko、Ishizu Akihiro、Watanabe Masafumi
  • Journal Title: Respiratory Investigation Volume: 59 Issue: 3 Pages: 372-376
  • DOI: 10.1016/j.resinv.2020.12.002
  • Peer Reviewed
• [Journal Article] Neutrophil fixation protocols suitable for substrates to detect anti-neutrophil cytoplasmic antibodies by indirect immunofluorescence (2021)
  • Author(s): Nishibata Yuka、Matsuzawa Shun、Satomura Yosuke、Ohtsuka Takeshi、Kuhara Motoki、Masuda Sakiko、Tomaru Utano、Ishizu Akihiro
  • Journal Title: Pathology - Research and Practice Volume: 228 Pages: 153661-153661
  • DOI: 10.1016/j.prp.2021.153661
  • NAID: 120007171671
  • Peer Reviewed / Open Access
• [Presentation] 多発血管炎性肉芽腫症(GPA)の自然退縮における,好中球細胞外トラップ(NETs)分解について 当院で経験した1症例を通して (2021)
  • Author(s): 太田 啓貴, 佐藤 千紗, 五十嵐 朗, 邨野 浩義, 梁 秀鼎, 町田 浩祥, 佐藤 建人, 中野 寛之, 根本 貴子, 西脇 道子, 木村 友美, 山内 啓子, 佐藤 正道, 井上 純人, 渡辺 昌文, 益田 紗季子, 石津 明洋
  • Organizer: 日本呼吸器学会
• [Presentation] myosin light chain6を認議する抗好中球細胞外トラップ(NETs)抗体はNETs分解阻害活性を持つ (2021)
  • Author(s): 西端友香, 益田紗季子, 外丸詩野, 石津明洋
  • Organizer: 日本リウマチ学会総会・学術集会
• [Presentation] 低酸素環境が好中球細胞外トラップ形成へ与える影響 (2021)
  • Author(s): 益田紗季子, 西端友香, 田中敏, 外丸詩野, 辻野一三, 石津明洋
  • Organizer: 日本病理学会