Development of inhibitors of osteoclast differentiation with an anti-citrullinated protein monoclonal antibody

• Project/Area Number: 19K08908
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 54020:Connective tissue disease and allergy-related
• Research Institution: Hiroshima University
• Principal Investigator: SUGIYAMA EIJI 広島大学, 病院(医), 名誉教授 (70179167)
• Co-Investigator(Kenkyū-buntansha): 小澤 龍彦 富山大学, 学術研究部医学系, 准教授 (10432105) ; 應原 一久 広島大学, 医系科学研究科(歯), 助教 (80550425)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2021: ¥390,000 (Direct Cost: ¥300,000、Indirect Cost: ¥90,000); Fiscal Year 2020: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2019: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 関節リウマチ / 抗シトルリン化蛋白抗体 / SKGマウス / モノクローナル抗体 / シトルリン化フィブリノーゲン / LPS / 滑膜細胞 / CXCL10 / 抗シトルリン化ペプチド抗体 / ＳＫＧマウス / 破骨細胞

Outline of Research at the Start

関節リウマチ（RA）は自己免疫性の慢性炎症性疾患で、その原因因子として遺伝的要因、環境的要因が明らかになっている。その病態としては、シトルリン化タンパク質に対する自己抗体（抗シトルリン化ペプチド抗体、ACPA)の増加がRAの発症、進展に深く関与している。申請者のグループはこれまでに、SKGマウスを用いた実験で、環境因子として歯周炎がRAを重篤化することをmRNA調節タンパク性因子HuR、補体C5aに着目し、明らかにしてきた。本研究ではACPAによる関節破壊メカニズムを破骨細胞分化に着目して明らかにするとともに、中和抗体として用いることで新たな治療薬としての可能性を検討する。

Outline of Final Research Achievements

We investigated the function of an anti-citrullinated protein monoclonal antibody (CCP-Ab1) obtained from rheumatoid arthritis patients, and found that the corresponding antigen is citrullinated fibrinogen. In addition, intravenous administration of CCP-Ab1 in arthritis-prone mouse model mice (SKG mice) resulted in worsening arthritis, progressive joint destruction, increased bone resorption, and increased osteoclasts at the boundary between bone and synovium. On the other hand, embryonic CCP-Ab1 did not show worsening of arthritis, suggesting that CCP-Ab1 acquires its virulence through repeated somatic mutations.

Academic Significance and Societal Importance of the Research Achievements

関節リウマチ診療では抗シトルリン化蛋白抗体（ACPA）は診断マーカー、関節破壊予測マーカーとして広く、使用しているが、その病原性に関しては不明である。今回の研究では関節リウマチから得られたACPAが強い病原性を有すること、その対応抗原はシトルリン化フィブリノーゲンであることを明らかにした。この実験結果はACPAそのものを治療ターゲットとして応用できることを示唆しており、新たなリウマチ治療の開発に貢献するものと思われる。

Research Products

• [Journal Article] The Lipopolysaccharide Mutant Re-LPS Is a Useful Tool for Detecting LPS Contamination in Rheumatoid Synovial Cell Cultures (2021)
  • Author(s): Kohno Hiroki、Ouhara Kazuhisa、Mokuda Sho、Tokunaga Tadahiro、Sugimoto Tomohiro、Watanabe Hirofumi、Ishitoku Michinori、Yoshida Yusuke、Mizuno Noriyoshi、Ozawa Tatsuhiko、Kawataka Masatoshi、Hirata Shintaro、Kishi Hiroyuki、Sugiyama Eiji
  • Journal Title: Pathobiology Volume: 89 Issue: 2 Pages: 92-100
  • DOI: 10.1159/000520022
  • Peer Reviewed
• [Journal Article] Nailfold capillaries and myositis specific antibodies in anti-melanoma differentiation-associated gene 5 antibody-positive dermatomyositis (2021)
  • Author(s): Sugimoto, T. Mokuda, S. Kohno, H. Ishitoku, M. Araki, K. Watanabe, H. Tokunaga, T. Yoshida, Y. Hirata, S. Sugiyama, E.
  • Journal Title: Rheumatology (Oxford) Volume: - Issue: 5 Pages: 2006-2015
  • DOI: 10.1093/rheumatology/keab681
  • Peer Reviewed
• [Journal Article] Angiotensin-converting Enzyme 2, a SARS-CoV-2 Receptor, Is Upregulated by Interleukin 6 through STAT3 Signaling in Synovial Tissues. (2020)
  • Author(s): Mokuda S, Tokunaga T, Masumoto J, Sugiyama E.
  • Journal Title: J Rheumatol. Volume: 47 Issue: 10 Pages: 2553-2558
  • DOI: 10.3899/jrheum.200547
  • Peer Reviewed / Open Access
• [Journal Article] Physiologic Target, Molecular Evolution, and Pathogenic Functions of a Monoclonal Anti-Citrullinated Protein Antibody Obtained From a Patient With Rheumatoid Arthritis. (2020)
  • Author(s): Ozawa T, Ouhara K, Tsuda R, Munenaga S, Kurihara H, Kohno H, Hamana H, Kobayashi E, Taki H, Tobe K, Sugiyama E, Muraguchi A, Kishi H.
  • Journal Title: Arthritis Rheumatol. Volume: 72 Issue: 12 Pages: 2040-2049
  • DOI: 10.1002/art.41426
  • Peer Reviewed / Open Access
• [Journal Article] Activin A Expressed in Rheumatoid Synovial Cells Downregulates TNFα-Induced CXCL10 Expression and Osteoclastogenesis (2020)
  • Author(s): Kuranobu T, Mokuda S, Oi K, Tokunaga T, Yukawa K, Kohno H, Yoshida Y, Hirata S, Sugiyama E
  • Journal Title: Pathobiology Volume: - Issue: 3 Pages: 198-207
  • DOI: 10.1159/000506260
  • Peer Reviewed
• [Journal Article] Effect of Porphyromonas gingivalis infection on gut dysbiosis and resultant arthritis exacerbation in mouse model (2020)
  • Author(s): Hamamoto Yuta、Ouhara Kazuhisa、Munenaga Syuichi、Shoji Mikio、Ozawa Tatsuhiko、Hisatsune Jyunzo、Kado Isamu、Kajiya Mikihito、Matsuda Shinji、Kawai Toshihisa、Mizuno Noriyoshi、Fujita Tsuyoshi、Hirata Shintaro、Tanimoto Kotaro、Nakayama Koji、Kishi Hiroyuki、Sugiyama Eiji、Kurihara Hidemi
  • Journal Title: Arthritis Research & Therapy Volume: 22 Issue: 1 Pages: 249-249
  • DOI: 10.1186/s13075-020-02348-z
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] TGFβ1 Regulates Human RANKL-Induced Osteoclastogenesis via Suppression of NFATc1 Expression (2020)
  • Author(s): Tokunaga T, Mokuda S, Kohno H, Yukawa K, Kuranobu T, Oi K, Yoshida Y, Hirata S, Sugiyama E
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 3 Pages: 800-800
  • DOI: 10.3390/ijms21030800
  • Peer Reviewed / Open Access
• [Journal Article] Serum CXCL10 levels are associated with better responses to abatacept treatment of rheumatoid arthritis. (2019)
  • Author(s): Yukawa K, Mokuda S, Kohno H, Oi K, Kuranobu T, Tokunaga T, Yoshida Y, Yamana J, Iwahashi M, Hirata S, Yamana S, Sugiyama E.
  • Journal Title: Clin Exp Rheumatol. Volume: -
  • Peer Reviewed
• [Presentation] Usefulness of nailfold video-capillaroscopy for diagnosing patients positive for anti-centromere antibody. (2021)
  • Author(s): Tomohiro Sugimoto, Hiroki Kobayashi, Harumichi Suma, Yohei Hosokawa, Michinori Ishitoku, Hiroki Kohno, Hirofumi Watanabe, Tadahiro Tokunaga, Shinpei Ohnari, Yusuke Yoshida, Sho Mokuda, Shintaro Hirata, Eiji Sugiyama
  • Organizer: 第６５回日本リウマチ学会総会・学術集会
• [Presentation] Angiotensin-converting enzyme 2, a SARS-CoV-2 receptor, is upregulated by IL-6 via STAT3 signaling in synovial fibroblasts and lymphocytes. (2021)
  • Author(s): Sho Mokuda, Tadahiro Tokunaga, Junya Masumoto, Eiji Sugiyama
  • Organizer: 第６５回日本リウマチ学会総会・学術集会
• [Presentation] リウマチ滑膜細胞のTLR4活性化研究におけるLPSのコンタミネーションの検出に、変異型LPS・Re-LPSは有用である (2021)
  • Author(s): 河野紘輝、應原一久、徳永忠浩、茂久田翔、水野智仁、小澤龍彦、川高正聖、平田信太郎、岸裕幸、杉山英二
  • Organizer: 第３６回日本臨床リウマチ学会