Tissue homeostasis of intervertebral disc

• Project/Area Number: 15K10420
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Keio University
• Principal Investigator: Fujita Nobuyuki 慶應義塾大学, 医学部(信濃町), 講師 (30348685)
• Co-Investigator(Kenkyū-buntansha): 松本 守雄 慶應義塾大学, 医学部(信濃町), 教授 (40209656)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 椎間板 / 髄核 / 繊維輪 / 酸化ストレス / 細胞株 / 椎間板変性 / 小胞体ストレス / JAK/STAT / FSTL1 / 脊索腫

Outline of Final Research Achievements

We show that a chordoma-derived cell line U-CH1-N cells highly express the nucleus pulposus (NP) marker genes, including T, KRT19, and CD24. These observations were further confirmed by immunocytochemistry and flow cytometry. Reporter analyses showed that transcriptional activity of T was enhanced in U-CH1-N cells. Chondrogenic capacity of U-CH1-N cells was verified by evaluating the expression of ECM genes and Alcian blue staining. Of note, we found that proliferation and synthesis of chondrogenic ECM proteins were largely dependent on T in U-CH1-N cells. In accordance, knockdown of the T transcripts suppressed the expression of PCNA, a gene essential for DNA replication, and SOX5 and SOX6, the master regulators of chondrogenesis. On the other hand, the CD24-silenced cells showed no reduction in the mRNA expression level of the chondrogenic ECM genes. These results suggest that U-CH1-N shares important biological properties with notochordal NP cells.

Research Products

• [Journal Article] The unfolded protein response mediated by PERK is casually related to the pathogenesis of intervertebral disc degeneration. (2017)
  • Author(s): T. Fujii, N. Fujita, S. Suzuki, K. Watanabe, T. Tsuji, K. Ishii, T. Takaki, K. Umezawa, T. Miyamoto, K. Horiuchi, M. Matsumoto, M. Nakamura
  • Journal Title: Journal of Orthopaedic Research Volume: - Issue: 5 Pages: 1334-1345
  • DOI: 10.1002/jor.23787
  • Peer Reviewed
• [Journal Article] Potential Involvement of the IL-6/JAK/STAT3 Pathway in the Pathogenesis of Intervertebral Disc Degeneration. (2017)
  • Author(s): Suzuki S, Fujita N, Fujii T, Watanabe K, Yagi M, Tsuji T, Ishii K, Miyamoto T, Horiuchi K, Nakamura M, Matsumoto M.
  • Journal Title: Spine (Phila Pa 1976) Volume: 42 Issue: 14 Pages: E817-E824
  • DOI: 10.1097/brs.0000000000001982
  • Peer Reviewed
• [Journal Article] Potential Involvement of the IL-6/JAK/STAT3 Pathway in the Pathogenesis of Intervertebral Disc Degeneration (2016)
  • Author(s): Suzuki S, Fujita N, Fujii T, Watanabe K, Yagi M, Tsuji T, Ishii K, Miyamoto T, Horiuchi K, Nakamura M, Matsumoto M
  • Journal Title: Spine Volume: 印刷中
  • Peer Reviewed
• [Journal Article] Chordoma-derived cell line U-CH1-N recapitulates the biological properties of notochordal nucleus pulposus cells (2016)
  • Author(s): Fujita N, Suzuki S, Watanabe K, Ishii K, Watanabe R, Shimoda M, Takubo K, Tsuji T, Toyama Y, Miyamoto T, Horiuchi K, Nakamura M, Matsumoto M
  • Journal Title: J Orthop Res Volume: ３４ Issue: 8 Pages: 1341-50
  • DOI: 10.1002/jor.23320
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Excessive reactive oxygen species are therapeutic targets for intervertebral disc degeneration. (2017)
  • Author(s): Fujita N, Suzuki S, Fujii T, Watanabe K, Ishii K, Tsuji T, Miyamoto T, Horiuchi K, Matsumoto M, Nakamura M.
  • Organizer: 30th annual meeting of Japanese Society of Cartilage Metabolism
  • Invited
• [Presentation] Potential Involvement of unfolded protein response in the pathogenesis of intervertbral disc degeneration. (2017)
  • Author(s): Fujita N, Fujii T.
  • Organizer: ORS PSRS 4th International Spine Research Symposium
  • Int'l Joint Research
• [Presentation] Molecular mechanisms of intervertebral disc degeneration. (2017)
  • Author(s): Fujita N.
  • Organizer: 5th International Spine symposium of Four Universities
  • Invited
• [Presentation] 椎間板変性に酸化ストレスが関与している (2017)
  • Author(s): 藤田順之
  • Organizer: 第35回日本骨代謝学会
  • Invited
• [Presentation] 椎間板変性に対する抗酸化剤の効果. (2017)
  • Author(s): 藤田順之
  • Organizer: 第20回日本低侵襲脊椎外科学会
  • Invited
• [Presentation] 腰痛に対する抗酸化治療 (2017)
  • Author(s): 藤田順之
  • Organizer: 第25回日本腰痛学会
  • Invited
• [Patent(Industrial Property Rights)] 小胞体ストレスを標的とする椎間板変性症対する治療薬 (2017)
  • Inventor(s): 藤田順之、藤井武、松本守雄、中村雅也
  • Industrial Property Rights Holder: 藤田順之、藤井武、松本守雄、中村雅也
  • Industrial Property Rights Type: 特許
  • Filing Date: 2017
• [Patent(Industrial Property Rights)] 椎間板変性症に対する治療薬ＪＡＫ‐ＳＴＡＴ３阻害剤 (2017)
  • Inventor(s): 藤田順之、鈴木悟士、松本守雄、中村雅也
  • Industrial Property Rights Holder: 藤田順之、鈴木悟士、松本守雄、中村雅也
  • Industrial Property Rights Type: 特許
  • Filing Date: 2017