膜電位操作回路による生体骨構築のための基盤研究－光照射による骨リモデリング制御－

• Project Area: Synthetic biology for the comprehension of biomolecular networks
• Project/Area Number: 24119505
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Complex systems
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: 納富 拓也 東京医科歯科大学, 難治疾患研究所, 特任講師 (70542249)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥14,040,000 (Direct Cost: ¥10,800,000、Indirect Cost: ¥3,240,000); Fiscal Year 2013: ¥7,020,000 (Direct Cost: ¥5,400,000、Indirect Cost: ¥1,620,000); Fiscal Year 2012: ¥7,020,000 (Direct Cost: ¥5,400,000、Indirect Cost: ¥1,620,000)
• Keywords: 骨代謝 / 破骨細胞 / 骨芽細胞 / 光 / 膜電位

Research Abstract

超高齢化社会を迎えて、骨粗鬆症を代表とする骨疾患は増え続けている。この治療・改善には、骨リモデリング機構を明らかにするとともに、そのコントロールが必要である。光活性化型分子を用いて、骨芽細胞・破骨細胞に膜電位操作回路を導入することに成功した。本研究では、創出した膜電位操作回路導入骨芽細胞・破骨細胞を用いて、骨リモデリング制御により、生体内において骨構築をおこなうことを目的とした。
本年度では、マウス生体内において、膜電位操作を介した骨リモデリング制御を通して骨構築をおこなうことを目的とした。確立した膜電位操作回路を用いて、生体内骨構築を試みた。頭頂骨由来の骨芽細胞に膜電位操作回路を導入して、マトリゲルとともにマウス頭頂骨欠損部に埋め込んだところ、細胞群の欠損部位における定着が確認された。透明プラスチックを通して光照射による膜電位操作をおこなうと、8週間後に骨欠損部の骨量回復が対照群に比べて顕著に認められた。この結果から、本研究で開発した膜電位操作回路の生体内における適用性が確認された。また、膜電位操作による細胞機能コントロールについても検討をすすめた。分子の細胞内輸送に焦点をあてて検討をおこなったところ、骨芽細胞において、破骨細胞形成因子の一つであるRANKLの細胞内輸送と膜電位変動の関係が明らかとなった。その細胞内輸送には、多様なイオンチャネルが関与しており、特に電位依存性のカルシウムチャネルの役割を確認した。これらの結果から、非興奮性細胞における膜電位変動の重要性が明らかとなった。

Current Status of Research Progress

Reason

25年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

25年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Migration linked to FUCCI-indicated cell cycle is controlled by PTH and mechanical stress (2014)
  • Author(s): Jumpei Shirakawa, Yoichi Ezura, Shuichi Moriya, Makiri Kawasaki, Takayuki Yamada, Takuya Notomi, Tetsuya Nakamoto, Tadayoshi Hayata, Atsushi Miyawaki, Ken Omura, Masaki Noda
  • Journal Title: J Cell Physiol Volume: -
  • Peer Reviewed
• [Journal Article] Stability of mRNA influences osteoporotic bone mass via Cnot3 (2014)
  • Author(s): Chiho Watanabe, Masahiro Morita, Yoichi Ezura, Tetsuya Nakamoto, Tadayoshi Hayata, Chisato Kikuguchi, Li Xue, Yasuhiro Kobayashi, Naoyuki Takahashi, Takuya Notomi, Keiji Moriyama, Tadashi Yamamoto, Masaki Noda
  • Journal Title: Proc Natl Acad Sci U S A Volume: 111 Pages: 2692-7
  • Peer Reviewed
• [Journal Article] Osteoblastic differentiation enhances expression of TRPV4 that is required for calcium oscillation induced by mechanical force (2013)
  • Author(s): Takafumi Suzuki, Takuya Notomi (Corresponding author), Daizuke Miyajima, Fumitaka Mizoguchi, Tadayoshi Hayata, Tetsuya Nakamoto, Ryo Hanyu, Paksinee Kamolratanakul, Atsuko Mizuno, Makoto Suzuki, Yoichi Ezura, Yuichi Izumi, Masaki Noda
  • Journal Title: Bone Volume: 54 Pages: 172-8
  • Peer Reviewed
• [Journal Article] Nck1 deficiency accelerates unloading-induced bone loss (2013)
  • Author(s): Aryal Smiriti, Kentaro Miyai, Yoichi Ezura, Tadayoshi Hayata, Takuya Notomi, Tetsuya Nakamoto, Tony Pawson, Masaki Noda
  • Journal Title: J. Cell. Physiol. Volume: 228 Pages: 1397-1403
  • Peer Reviewed
• [Journal Article] 骨研究フロンティア-基礎から臨床まで- 骨形成・骨吸収における関連因子の働き (2013)
  • Author(s): 野田政樹、江面陽一、早田匡芳、守屋秀一、白川純平、Smiriti Arayal、納富拓也、渡辺千穂、長尾雅史、羽生亮
  • Journal Title: 日本臨牀 Volume: 71 Pages: 161-166
• [Journal Article] Identification of two pore channel 2 as a novel regulator of osteoclastogenesis (2012)
  • Author(s): Takuya Notomi
  • Journal Title: Journal of Biological Chemistry Volume: 287(42) Issue: 42 Pages: 35057-64
  • DOI: 10.1074/jbc.m111.328930
  • Peer Reviewed
• [Journal Article] Profilin1 regulates sternum development and endochondral bone formation (2012)
  • Author(s): Daisuke Miyajima
  • Journal Title: J. Biol. Chem. Volume: 287 Pages: 33545-53
  • Peer Reviewed
• [Journal Article] Anabolic action of parathyroid hormone regulated by the beta2-adrenergic receptor (2012)
  • Author(s): Hanyu, R.
  • Journal Title: Proc Natl Acad Sci U S A Volume: 109 Issue: 19 Pages: 7433-7438
  • DOI: 10.1073/pnas.1109036109
  • Peer Reviewed
• [Journal Article] CIZ/NMP4 is expressed in B16 melanoma and forms a positive feedback loop with RANKL to promote migration of the melanoma cells (2012)
  • Author(s): Sakuma, T., Nakamoto, T., Hemmi, H., Kitazawa, S., Kitazawa, R., Notomi, T., Hayata, T., Ezura, Y., Amagasa, T., and Noda, M
  • Journal Title: Journal of cellular physiology Volume: 227 Issue: 7 Pages: 2807-2812
  • DOI: 10.1002/jcp.24066
  • Peer Reviewed
• [Presentation] Lysosomal Calcium Channel, TPC2, Regulates Osteoclastogenesis via Generation of Intracellular Ca2+ Response and Subsequent NFATc1 Localization: A Novel Mechanism of Osteoclastic Ca2+ Signaling (2012)
  • Author(s): Takuya Notomi
  • Organizer: ASBMR 2012
  • Place of Presentation: Minneapolis, Minnesota, USA
  • Year and Date: 2012-10-12
• [Presentation] Parathyroid Hormone Stimulates Tob1 Expression in Osteoblastic Cells in vitro and in vivo (2012)
  • Author(s): Shuichi Moriya
  • Organizer: ASBMR 2012
  • Place of Presentation: Minneapolis, Minnesota, USA
  • Year and Date: 2012-10-12
• [Presentation] PTH Enhances Mechanical Stress-induced Osteoblast Proliferation in Calvarial Derived Osteoblasts via Up-regulation of CyclinD1 Expression (2012)
  • Author(s): Shirakawa Jumpei
  • Organizer: ASBMR 2012
  • Place of Presentation: Minneapolis, Minnesota, USA
  • Year and Date: 2012-10-12
• [Presentation] Cnot3 (Ccr4-not complex subunit3), a Regulator of mRNA Stability, Regulates Bone Mass and Gene Expression Related to Osteoclast Formation (2012)
  • Author(s): Chiho Watanabe
  • Organizer: ASBMR 2012
  • Place of Presentation: Minneapolis, Minnesota, USA
  • Year and Date: 2012-10-12
• [Presentation] Identification of Signature Genes Selectively Expressed in Mesenchymal Stem Cells Derived from Synovial Joint Tissues (2012)
  • Author(s): Yoichi Ezura
  • Organizer: ASBMR 2012
  • Place of Presentation: Minneapolis, Minnesota, USA
  • Year and Date: 2012-10-12
• [Presentation] Nck, an Actin Cytoskeleton Modulator, Controls Expression of Osteocytic Genes, Phosphate Homeostasis by Regulating FGF 23 Expression in Bone and Maintains Bone Mass (2012)
  • Author(s): Smriti Aryal
  • Organizer: ASBMR 2012
  • Place of Presentation: Minneapolis, Minnesota, USA
  • Year and Date: 2012-10-12
• [Presentation] TGF-β1 Decreases Ift88 Expression in Chondrocytic Cell Line ATDC5 (2012)
  • Author(s): Makiri Kawasaki
  • Organizer: ASBMR 2012
  • Place of Presentation: Minneapolis, Minnesota, USA
  • Year and Date: 2012-10-12
• [Presentation] Two Pore Channel 2を介したリソソーム由来Ca2+による新たな破骨細胞分化制御機構 (2012)
  • Author(s): 納富拓也
  • Organizer: 第30回 日本骨代謝学会
  • Place of Presentation: 東京
  • Year and Date: 2012-07-19
• [Presentation] 一次繊毛タンパクBbs3は骨代謝に関与する (2012)
  • Author(s): 川崎真希理
  • Organizer: 第30回 日本骨代謝学会
  • Place of Presentation: 東京
  • Year and Date: 2012-07-19
• [Presentation] 骨量制御の新転写後性分子機構：mRNA deadenylase であるCcr4-not complex構成因子Cnot3の欠失による高回転型の骨量減少の解析 (2012)
  • Author(s): 渡辺千穂
  • Organizer: 第30回 日本骨代謝学会
  • Place of Presentation: 東京
  • Year and Date: 2012-07-19
• [Presentation] 骨芽細胞におけるGタンパク質共役型受容体(GPCR)によるRANKL制御の検討 (2012)
  • Author(s): 守屋秀一
  • Organizer: 第30回 日本骨代謝学会
  • Place of Presentation: 東京
  • Year and Date: 2012-07-19
• [Presentation] ヒト骨髄および滑膜由来間葉系細胞において異なるCpGメチル化を示す遺伝子群の探索と骨軟骨細胞分化の制御に関わる転写因子群の抽出：RUNX2およびRUNX3, D+X5, ALX4遺伝子 (2012)
  • Author(s): 江面陽一
  • Organizer: 第30回 日本骨代謝学会
  • Place of Presentation: 東京
  • Year and Date: 2012-07-19
• [Presentation] Depolarizing Membrane Potential by PTH and VD3 Regulates RANKL-intracellular Transportation; A Novel Mechanism of PTH- and VD3-induced Osteoclastogenesis
  • Author(s): Takuya Notomi, Miyuki Kuno, Yoichi Ezura, Masaki Noda
  • Organizer: ASBMR 2013
  • Place of Presentation: メリーランド州、米国
• [Presentation] Genes significantly highly expressed in synovium derived stromal cells than in bone marrow derived cells are conserved both in mouse and human, and may contribute to higher potential for chondrogenic differentiation
  • Author(s): Yoichi Ezura, Tadayoshi Hayata, Takuya Notomi, Ichiro Sekiya, Masaki Noda
  • Organizer: ASBMR 2013
  • Place of Presentation: メリーランド州、米国
• [Presentation] PTH Additively Enhances The Mechanical Stress-induced Proliferation of Calvarial Osteoblasts
  • Author(s): Jumpei Shirakawa, Yoichi Ezura, Makiri Kawasaki, Takayuki Yamada, Shuichi Moriya, Takuya Notomi, Tadayoshi Hayata, Ken Omura, Masaki Noda
  • Organizer: ASBMR 2013
  • Place of Presentation: メリーランド州、米国
• [Presentation] TGF-β1 inhibits maturation of chondrogenic cell line ATDC5 by impeding canonical hedgehog signaling through direct down-regulation of ciliary component gene Ift88
  • Author(s): Makiri Kawasaki, Tetsuya Nakamoto, Takuya Notomi, Tadayoshi Hayata, Yoichi Ezura, Masaki Noda
  • Organizer: ASBMR 2013
  • Place of Presentation: メリーランド州、米国
• [Presentation] Tob1, a BMP repressor, is activated by parathyroid hormone in osteoblasts in vitro and in vivo and reciprocally regulates PTH signaling
  • Author(s): Shuichi Moriya, Tadayoshi Hayata, Jumpei Shirakawa, Makiri Kawasaki, Takuya Notomi, Yoichi Ezura, Kazuo Kaneko, Masaki Noda
  • Organizer: ASBMR 2013
  • Place of Presentation: メリーランド州、米国
• [Presentation] β2Adrenergic Receptor agonist suppresses BMP-induced osteoblastic differentiation in MC3T3E-1 cells while epinephrine modulates it differently
  • Author(s): Takayuki Yamada, Tadayoshi Hayata, Takuya Notomi, Yoichi Ezura, Kiyoshi Harada, Masaki Noda
  • Organizer: ASBMR 2013
  • Place of Presentation: メリーランド州、米国
• [Presentation] 破骨細胞分化段階における細胞内Ca2+動態とイオンチャネルの役割
  • Author(s): 納富拓也、江面陽一、野田政樹
  • Organizer: 日本薬理学会年会 2014
  • Place of Presentation: 仙台国際センター
  • Invited
• [Presentation] 滑膜・半月板および靭帯由来間葉系幹細胞における選択的発現遺伝子の同定
  • Author(s): 江面陽一、早田匡芳、中元哲也、納富拓也、関谷一郎、宗田大、野田政樹
  • Organizer: 第31回 日本骨代謝学会
  • Place of Presentation: 神戸国際会議場