Elucidation of regulatory mechanisms of skeletal via Smad4

• Project/Area Number: 17K11026
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Saitama Medical University
• Principal Investigator: Tsukamoto Sho 埼玉医科大学, 医学部, 助教 (20707658)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 骨形成 / 骨系統疾患 / 骨粗鬆症 / シグナル伝達 / 骨代謝 / 骨・軟骨代謝学 / 内軟骨性骨化 / 骨、軟骨代謝学

Outline of Final Research Achievements

Members of the transforming growth factor-β (TGF-β) family regulate skeletal formation through activating Smad transcription factors. Smad4 is a critical coactivator essential for the Smad-dependent intracellular signaling. We established Smad4 conditional knockout (Smad4 cKO) mice under the control of tamoxifen. The Smad4 cKO mice showed an increase in bone mass in trabecular bone. They also showed an increase in β-catenin positive osteoblasts in spongiosa. The analysis of the Wnt signaling related genes revealed that expression of Wnt7b was increased in Smad4 cKO mice. These results suggest that the inhibition of Smad4-dependent signaling enhances bone formation through increased expression of Wnt7b.

Academic Significance and Societal Importance of the Research Achievements

本研究から、出生後におけるSmad4を介したシグナルは、骨形成を抑制している可能性を見出した。さらに、TGF-βファミリーと骨形成を強力に亢進するWntシグナルの新たなクロストークの発見は、骨粗鬆症をはじめとした骨代謝関連の疾患の新たな治療標的となる可能性がある。

Research Products

• [Journal Article] Fibrodysplasia ossificans progressiva: Review and research activities in Japan (2020)
  • Author(s): Haga Nobuhiko、Nakashima Yasuharu、Kitoh Hiroshi、Kamizono Junji、Katagiri Takenobu、Saijo Hideto、Tsukamoto Sho、Shinoda Yusuke、Sawada Ryoko、Nakahara Yasuo
  • Journal Title: Pediatrics International Volume: 62 Issue: 1 Pages: 3
  • DOI: 10.1111/ped.14065
  • Peer Reviewed / Open Access
• [Journal Article] Discovery of Heterotopic Bone-Inducing Activity in Hard Tissues and the TGF-β Superfamily (2018)
  • Author(s): Katagiri Takenobu、Tsukamoto Sho、Nakachi Yutaka、Kuratani Mai
  • Journal Title: International Journal of Molecular Sciences Volume: 19 Issue: 11 Pages: 3586-3586
  • DOI: 10.3390/ijms19113586
  • Peer Reviewed / Open Access
• [Journal Article] Recent Topics in Fibrodysplasia Ossificans Progressiva (2018)
  • Author(s): Katagiri Takenobu、Tsukamoto Sho、Nakachi Yutaka、Kuratani Mai
  • Journal Title: Endocrinology and Metabolism Volume: 33 Issue: 3 Pages: 331-331
  • DOI: 10.3803/enm.2018.33.3.331
  • Peer Reviewed / Open Access
• [Journal Article] Effects of FKBP12 and type II BMP receptors on signal transduction by ALK2 activating mutations associated with genetic disorders (2018)
  • Author(s): Machiya Aiko、Tsukamoto Sho、Ohte Satoshi、Kuratani Mai、Fujimoto Mai、Kumagai Keigo、Osawa Kenji、Suda Naoto、Bullock Alex N.、Katagiri Takenobu
  • Journal Title: Bone Volume: 111 Pages: 101-108
  • DOI: 10.1016/j.bone.2018.03.015
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Heterotopic bone induction via BMP signaling: Potential therapeutic targets for fibrodysplasia ossificans progressiva (2018)
  • Author(s): Katagiri Takenobu、Tsukamoto Sho、Kuratani Mai
  • Journal Title: Bone Volume: 109 Pages: 241-250
  • DOI: 10.1016/j.bone.2017.07.024
  • Peer Reviewed / Open Access
• [Presentation] Wnt7b expressed by hypertrophic chondrocytes is a critical inducer of bone formation during endochondral ossification (2019)
  • Author(s): Tsukamoto S, Kuratani M, Sekine N, Okubo M, Tanaka S, Jimi E, Oda H, Katagiri T
  • Organizer: ASBMR 2019 Annual meeting
  • Int'l Joint Research
• [Presentation] A crosstalk between Wnt and TGF-β/BMP families regulates endochondral bone formation (2019)
  • Author(s): Tsukamoto S, Kuratani M, Sekine N, Okubo M, Tanaka S, Jimi E, Oda H, Katagiri T
  • Organizer: Gordon Research Conference on Wnt Signaling
  • Int'l Joint Research
• [Presentation] 内軟骨性骨化におけるBMPとWntの新たなクロストーク (2019)
  • Author(s): 塚本翔、倉谷麻衣、関根典子、大久保美里、田中伸哉、自見英治郎、織田弘美、片桐岳信
  • Organizer: 第5回日本骨免疫学会
• [Presentation] Wnt7bは分化成熟した軟骨細胞が発現する内軟骨性骨化における骨形成促進因子である (2019)
  • Author(s): 塚本翔、倉谷麻衣、田中伸哉、自見英治郎、織田弘美、片桐岳信
  • Organizer: 第37回日本骨代謝学会学術集会
  • Int'l Joint Research
• [Presentation] Identiffication of a critical amino acid residue in ALK2 for the binding and inhibition by the anti-ALK2 blocking antibody (2019)
  • Author(s): Katagiri T, Tsukamoto S, Kuratani M, Tsuji S, Takaishi K, Kunikata T, Nakamura K, Kawaguchi Y, Hasegawa J
  • Organizer: ASBMR 2019 Annual meeting
  • Int'l Joint Research
• [Presentation] 抗ALK2阻害抗体の結合と阻害活性に重要なALK2細胞外領域アミノ酸残基の同定 (2019)
  • Author(s): 片桐岳信、塚本翔、倉谷麻衣
  • Organizer: 第37回日本骨代謝学会学術集会
• [Presentation] Identiffication of a critical amino acid residue in ALK2 for the binding and inhibition by the anti-ALK2 blocking antibody (2019)
  • Author(s): Katagiri T, Tsukamoto S, Kuratani M, Tsuji S, Takaishi K, Kunikata T, Nakamura K, Kawaguchi Y, Hasegawa J
  • Organizer: FASEB SRC on TGF-β Superfamily
  • Int'l Joint Research
• [Presentation] A role of dimer formation of ALK2, a BMP typeI receptor in the activation of intracellular signaling (2019)
  • Author(s): Katagiri T, Tsuji S, Tsukamoto S, Kuratani M, Ohte S, Sekine N, Kumagai K, Osawa K, Takaishi K, Nakamura K, Kawaguchi Y, Hasegawa J
  • Organizer: Gordon Research Conference on Cartilage
  • Int'l Joint Research
• [Presentation] A novel crosstalk between TGF-β/BMP and Wnt families through Smad4 in endochondral ossification (2018)
  • Author(s): Tsukamoto S, Kuratani M, Sekine N, Okubo M, Nakachi Y, Tanaka S, Jimi E, Oda H, Katagiri T
  • Organizer: ASBMR 2018 Annual Meeting
  • Int'l Joint Research
• [Presentation] A novel crosstalk between TGF-β/BMP and Wnt families through Smad4 in endochondral ossification (2018)
  • Author(s): Tsukamoto S, Kuratani M, Sekine N, Okubo M, Nakachi Y, Tanaka S, Jimi E, Oda H, Katagiri T
  • Organizer: Mechanistic and Therapeutic Advances in Rare Skeletal Diseases Research Meeting [A meeting in affiliation with ASBMR 2018]
  • Int'l Joint Research
• [Presentation] A novel crosstalk between BMP and Wnt7b in postnatal endochondral ossification (2018)
  • Author(s): Tsukamoto S, Kuratani M, Sekine N, Okubo M, Nakachi Y, Tanaka S, Jimi E, Oda H, Katagiri, T
  • Organizer: 12th International BMP Conference
  • Int'l Joint Research
• [Presentation] A novel crosstalk between the TGF-β and Wnt families in endochondral ossification (2018)
  • Author(s): Tsukamoto S, Kuratani M, Sekine N, Okubo M, Nakachi Y, Katagiri T
  • Organizer: The 16th RCGM International Symposium of Academic Frontier
• [Presentation] TGF-βファミリーはWntシグナルを介して内軟骨性骨化を制御する (2018)
  • Author(s): 塚本翔、倉谷麻衣、田中伸哉、自見英治郎、織田弘美、片桐岳信
  • Organizer: 第36回日本骨代謝学会学術集会
• [Presentation] An antibody against ALK2 extracellular domain reveals a role of dimer formation for signal activation (2018)
  • Author(s): Katagiri T, Tsuji S, Tsukamoto S, Kuratani M, Ohte S, Takaishi K, Nakamura K, Kawaguchi Y, Hasegawa J
  • Organizer: ASBMR 2018 Annual Meeting
  • Int'l Joint Research
• [Presentation] Expression levels of BMP ligands are increased by muscle trauma (2018)
  • Author(s): Kuratani M, Tsukamoto S, Sekine N, Okubo M, Nakachi Y, Katagiri, T
  • Organizer: 12th International BMP Conference
  • Int'l Joint Research
• [Presentation] 進行性骨化性線維異形成症（FOP）モデルマウスを用いた抗ALK2抗体の解析 (2018)
  • Author(s): 片桐岳信、辻真之介、塚本翔、倉谷麻衣、大手聡、高石巨澄
  • Organizer: 第36回日本骨代謝学会学術集会
• [Presentation] Analysis of skeletal growth in Smad4 knockout mice (2017)
  • Author(s): Tsukamoto S, Sekine N, Kuratani M, Kumagai K, Tanaka S, Jimi E, Oda H, Katagiri T
  • Organizer: 2017 FASEB Science Research Conferences on The TGF-β Superfamily
  • Int'l Joint Research
• [Presentation] Development of blocking monoclonal antibodies against ALK2, which is a type I receptor for BMPs (2017)
  • Author(s): Katagiri T, Tsuji S, Tsukamoto S, Ohte S, Kumagai K, Osawa K, Takaishi K, Nakamura K, Kawaguchi Y, Hasegawa J
  • Organizer: 2017 FASEB Science Research Conferences on The TGF-β Superfamily
  • Int'l Joint Research
• [Presentation] Postnatal ablation of Smad4 enhances endochondral ossification in epiphyseal growth plate (2017)
  • Author(s): Tsukamoto S, Sekine N, Kuratani M, Kumagai K, Tanaka S, Jimi E, Oda H, Katagiri T
  • Organizer: 2017 ASBMR Annual Meeting
  • Int'l Joint Research
• [Presentation] Development of blocking monoclonal antibodies against ALK2, which is a type I receptor for BMPs (2017)
  • Author(s): Katagiri T, Tsuji S, Tsukamoto S, Ohte S, Kumagai K, Osawa K, Takaishi K, Nakamura K, Kawaguchi Y, Hasegawa J
  • Organizer: 2017 ASBMR Annual Meeting
  • Int'l Joint Research
• [Presentation] Analysis of skeletal phenotypes in Smad4 conditional knockout mice (2017)
  • Author(s): Tsukamoto S, Sekine N, Kuratani M, Kumagai K, Jimi E, Katagiri T
  • Organizer: The 65th Annual Meeting of Japanese Association for Dental Research
  • Int'l Joint Research
• [Presentation] Analysis of the TGF-β family ligands that are increased in regenerating skeletal muscle and induce BMP signaling via a mutant ALK2 associated with fibrodysplasia ossificans progressiva (2017)
  • Author(s): Kuratani M, Tsukamoto S, Katagiri T
  • Organizer: The 65th Annual Meeting of Japanese Association for Dental Research
  • Int'l Joint Research
• [Presentation] 転写共役因子Smad4は軟骨細胞分化を介して骨形成を制御する (2017)
  • Author(s): 塚本翔、関根典子、倉谷麻衣、熊谷桂吾、大久保美里、片桐岳信
  • Organizer: 第15回RCGMフロンティアシンポジウム
• [Presentation] BMP受容体ALK2を介した異所性骨化モデルの構築 (2017)
  • Author(s): 倉谷麻衣、塚本翔、関根典子、片桐岳信
  • Organizer: 第15回RCGMフロンティアシンポジウム
• [Presentation] TGF-βファミリーの軟骨細胞分化を介した骨形成への役割 (2017)
  • Author(s): 塚本翔
  • Organizer: 第14回彩の国骨フォーラム
• [Remarks] 埼玉医科大学ゲノム医学研究センター
  • URL: http://www.saitama-med.ac.jp/genome/