Analysis of cellular dynamics in arthritis and evaluation of drug mechanism

• Project/Area Number: 19K17906
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 54020:Connective tissue disease and allergy-related
• Research Institution: Osaka University
• Principal Investigator: Kikuta Junichi 大阪大学, 生命機能研究科, 准教授 (60710069)
• Project Period (FY): 2019-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2020: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2019: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 生体イメージング / 二光子励起顕微鏡 / 破骨細胞 / 関節破壊 / 分子標的治療薬 / 関節リウマチ / 免疫学

Outline of Research at the Start

本研究は、これまでノウハウを蓄積してきた生体二光子励起イメージング技術を駆使して、関節炎・関節破壊に関わる炎症細胞の遊走動態を可視化し、関節炎の発症機序を明らかにするとともに、関節リウマチ治療において臨床応用されている分子標的治療薬の生体内動態・薬効発現メカニズムを解明する。

Outline of Final Research Achievements

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by synovial joint inflammation and progressive bone destruction. In this study, we established an intravital imaging system to observe synovial tissues using a multiphoton microscopy and succeeded in visualizing pathological osteoclasts and their precursors at the pannus-bone interface. Combined with fluorescence-labeling of CTLA4-Ig, a biological agent used for the treatment of RA, we identified that CTLA4-Ig was distributed predominantly within the inflamed synovium and bound to CX3CR1+ macrophages and CD140a+ fibroblasts, but not to mature osteoclasts. Intravital synovial imaging system facilitates investigation of cellular dynamics in the pathogenesis of arthritis, and would thus be useful for evaluating drug mechanism and efficacy of biological agents within the synovial microenvironment in vivo.

Academic Significance and Societal Importance of the Research Achievements

本研究は、ライブイメージング技術の特徴を生かし、生体の炎症関節内における破骨細胞の動態や分子標的治療薬の体内動態を細胞レベルで時空間的に解明したもので、国際的にも独自性が高く、学術的に大きな意義がある。最先端のイメージング技術によって得られた基礎研究成果を臨床現場へ還元することにより、国内外のリウマチ医療へ貢献することができ、その社会的意義も極めて大きい。

Research Products

• [Journal Article] Development of an intravital imaging system for the synovial tissue reveals the dynamics of CTLA-4 Ig in vivo (2020)
  • Author(s): Hasegawa Tetsuo、Kikuta Junichi、Sudo Takao、Yamashita Erika、Seno Shigeto、Takeuchi Tsutomu、Ishii Masaru
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 13480-13480
  • DOI: 10.1038/s41598-020-70488-y
  • Peer Reviewed / Open Access
• [Journal Article] Local sympathetic neurons promote neutrophil egress from the bone marrow at the onset of acute inflammation (2020)
  • Author(s): Ao Tomoka、Kikuta Junichi、Sudo Takao、Uchida Yutaka、Kobayashi Kenta、Ishii Masaru
  • Journal Title: International Immunology Volume: 32 Issue: 11 Pages: 727-736
  • DOI: 10.1093/intimm/dxaa025
  • NAID: 120006952209
  • Peer Reviewed / Open Access
• [Journal Article] In vivo dynamic analysis of BMP-2-induced ectopic bone formation. (2020)
  • Author(s): Hashimoto K, Kaito T, Furuya M, Seno S, Okuzaki D, Kikuta J, Tsukazaki H, Matsuda H, Yoshikawa H, Ishii M.
  • Journal Title: Sci Rep. Volume: 10 Issue: 1 Pages: 4751-4751
  • DOI: 10.1038/s41598-020-61825-2
  • Peer Reviewed / Open Access
• [Journal Article] Intravital multiphoton microscopy as a novel tool in the field of immunopharmacology. (2020)
  • Author(s): Morimoto A, Kikuta J, Ishii M.
  • Journal Title: Pharmacol Ther. Volume: 206 Pages: 107429-107429
  • DOI: 10.1016/j.pharmthera.2019.107429
  • Peer Reviewed
• [Journal Article] Thrombomodulin induces anti-inflammatory effects by inhibiting the rolling adhesion of leukocytes in vivo. (2020)
  • Author(s): Nishizawa S, Kikuta J, Seno S, Kajiki M, Tsujita R, Mizuno H, Sudo T, Ao T, Matsuda H, Ishii M.
  • Journal Title: J Pharmacol Sci. Volume: 143 Issue: 1 Pages: 17-22
  • DOI: 10.1016/j.jphs.2020.01.001
  • Peer Reviewed / Open Access
• [Journal Article] Identification of a novel arthritis-associated osteoclast precursor macrophage regulated by FoxM1. (2019)
  • Author(s): Hasegawa T, Kikuta J, Sudo T, Matsuura Y, Matsui T, Simmons S, Ebina K, Hirao M, Okuzaki D, Yoshida Y, Hirao A, Kalinichenko VV, Yamaoka K, Takeuchi T, Ishii M.
  • Journal Title: Nat Immunol. Volume: 20 Issue: 12 Pages: 1631-1643
  • DOI: 10.1038/s41590-019-0526-7
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] In Vivo Multicolor Imaging with Fluorescent Probes Revealed the Dynamics and Function of Osteoclast Proton Pumps (2019)
  • Author(s): Minoshima Masafumi、Kikuta Junichi、Omori Yuta、Seno Shigeto、Suehara Riko、Maeda Hiroki、Matsuda Hideo、Ishii Masaru、Kikuchi Kazuya
  • Journal Title: ACS Central Science Volume: 5 Issue: 6 Pages: 1059-1066
  • DOI: 10.1021/acscentsci.9b00220
  • Peer Reviewed / Open Access
• [Presentation] 骨破壊・免疫炎症の生体イメージング (2020)
  • Author(s): 菊田順一, 石井優
  • Organizer: 第41回日本炎症・再生医学会
  • Invited
• [Presentation] 骨免疫イメージングアップデート (2020)
  • Author(s): 菊田順一, 石井優
  • Organizer: 第38回日本骨代謝学会学術集会
  • Invited
• [Presentation] 生体イメージングによる疾患・創薬研究 (2019)
  • Author(s): 菊田順一, 石井優
  • Organizer: 第63回日本リウマチ学会総会・学術集会
  • Invited
• [Presentation] In vivo visualization of immune cell dynamics and drug actions in rheumatic diseases. (2019)
  • Author(s): Kikuta J, Ishii M.
  • Organizer: EAGOR2019
  • Int'l Joint Research / Invited
• [Presentation] 生体試料を見る (2019)
  • Author(s): 菊田順一, 石井優
  • Organizer: 第37回日本骨代謝学会学術集会
  • Invited