Development of a novel osteoinductive factor using NELL1 proteins

• Project/Area Number: 26350959
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biomolecular chemistry
• Research Institution: Nagoya University
• Principal Investigator: Niimi Tomoaki 名古屋大学, 生命農学研究科, 講師 (30377791)
• Co-Investigator(Renkei-kenkyūsha): KURODA Shun'ichi 大阪大学, 産業科学研究所, 教授 (60263406)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 骨分化 / 細胞接着 / 再生医療 / 骨芽細胞

Outline of Final Research Achievements

The NELL1 gene was originally identified in craniosynostosis patients as being specifically unregulated within prematurely fusing sutures. Because of its potent osteoinductive activity, NELL1 protein may be useful for bone regeneration therapy. However, there is little knowledge regarding NELL1 receptors and NELL1-mediated signaling pathways. In this study, we demonstrate that NELL1 binds to cell surface proteoglycans through its thrombospondin-1 N-terminal (TSPN) domain. Major heparin-binding sites were identified on the three-dimensional structural model of the TSPN domain of NELL1. Mutant analysis of the heparin-binding sites indicated that the heparin-binding activity of the TSPN domain is involved in interaction of NELL1 with cell surface proteoglycans. We also found that NELL1 binds to Robo3 receptor through its EGF-like repeats.

Research Products

• [Journal Article] Mapping the heparin-binding site of the osteoinductive protein NELL1 by site-directed mutagenesis. (2015)
  • Author(s): Takahashi, K., Imai, A., Iijima, M., Yoshimoto, N., Maturana, A. D., Kuroda, S., and Niimi, T.
  • Journal Title: FEBS Letters Volume: 589 Issue: 24PartB Pages: 4026-4032
  • DOI: 10.1016/j.febslet.2015.11.032
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Oligomerization-induced conformational change in the C-terminal region of Nel-like molecule 1 (NELL1) protein is necessary for the efficient mediation of murine MC3T3-E1 cell adhesion and spreading (2014)
  • Author(s): Nakamura, Y., Hasebe, A., Takahashi, K., Iijima, M., Yoshimoto, N., Maturana, A. D., Ting, K., Kuroda, S., and Niimi T.
  • Journal Title: Journal of Biological Chemistry Volume: 289 Issue: 14 Pages: 9781-9794
  • DOI: 10.1074/jbc.m113.507020
  • Peer Reviewed / Open Access
• [Presentation] 骨分化誘導タンパク質NELL1のRoboファミリー受容体への結合能の解析 (2017)
  • Author(s): 山本菜央佳、新美友章
  • Organizer: 日本農芸化学会2017年度大会
  • Place of Presentation: 京都女子大学
  • Year and Date: 2017-03-17
• [Presentation] 骨分化誘導タンパク質NELL1のヘパリン結合部位の解析 (2015)
  • Author(s): 高橋謙嘉、今井杏理紗、新美友章
  • Organizer: 第38回日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2015-12-01
• [Presentation] 骨分化誘導タンパク質NELL1のヘパリン結合活性の役割 (2014)
  • Author(s): 今井杏理紗、黒田俊一、新美友章
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ橫浜（橫浜）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 骨形成蛋白質NELL1のインテグリンおよびヘパラン硫酸プロテオグリカンを介したシグナル伝達 (2014)
  • Author(s): 高橋謙嘉、黒田俊一、新美友章
  • Organizer: 第46回日本結合組織学会学術大会・第61回マトリックス研究会大会合同学術集会
  • Place of Presentation: ウインクあいち（名古屋）
  • Year and Date: 2014-06-05 – 2014-06-07