c-Srcリン酸化を介する転写因子NF-κBの活性化と骨吸収の制御

• Project/Area Number: 12137205
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Osaka University
• Principal Investigator: 米田 俊之 大阪大学, 大学院・歯学研究科, 教授 (80142313)
• Co-Investigator(Kenkyū-buntansha): 西村 理行 大阪大学, 大学院・歯学研究科, 助教授 (60294112) ; 平賀 徹 大阪大学, 大学院・歯学研究科, 講師 (70322170)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥51,900,000 (Direct Cost: ¥51,900,000); Fiscal Year 2004: ¥13,600,000 (Direct Cost: ¥13,600,000); Fiscal Year 2003: ¥13,500,000 (Direct Cost: ¥13,500,000); Fiscal Year 2002: ¥12,500,000 (Direct Cost: ¥12,500,000); Fiscal Year 2001: ¥12,300,000 (Direct Cost: ¥12,300,000)
• Keywords: 破骨細胞 / 骨吸収 / RANKL / c-Src / NFAT / c-Jun / JNK / アクチン / Chk / Csk

Research Abstract

1.In vivoでのNFATファミリー遺伝子の破骨細胞の分化・機能における役割
研究代表者らは、RANKLにより制御される転写因子として、NFATフアミリーが破骨細胞の分化制御において重要性な役割を演じていることを明らかにした。さらにNFATファミリーのin vivoにおける役割を明らかにするために、NFAT遺伝子の改変マウスを作成し、この遺伝子改変マウスの表現型を詳細に解析を行った。その結果、このトランスジェニックマウスでは、破骨細胞の形成ならびに骨吸収が著明に亢進していることが明らかとなった。
2.c-Srcの活性化機構の解明
破骨細胞の骨吸収活性制御に必須であるc-Srcの機能調節機構を明らかにするために、c-Src制御キナーゼCskおよびその調節因子Cbpの破骨細胞における関与を検討した。その結果、破骨細胞では、CskをLipid RaftにリクルートするCbpの発現が著しく低下しており、その結果、c-Src活性が高く維持されていることが明らかとなった。さらにアデノウィルスを用いたCbp導入実験により、Csk活性の促進、c-Src活性の低下、骨吸収の低下が確認され、この知見の正当性を強く支持できた。
3.TRAF6とNFATの相互関連に関する検討
研究代表者らは、NFATの発現および機能の活性化に関して細胞内シグナル伝達分子TRAF6が、密接に関与していることを見出した。さらに分子細胞生物学的アプローチにより、TRAF6とNFATの相互関係の解明を行った結果、TRAF6は、NFATの転写活性を促進させ、破骨細胞分化を促進していることが示唆された。

Research Products

• [Journal Article] CCAAT/enhancer binding protein β isoform, LIP, regulates commitment of osteoblasts and adipocytes (2005)
  • Author(s): Hata K
  • Journal Title: Mol Cell Biol 25 Pages: 1971-1979
• [Journal Article] A synthetic peptide fragment of human MEPE stimulates new bone formation in vitro and in vivo. (2004)
  • Author(s): Hayashibara T
  • Journal Title: J Bone Miner Res 19 Pages: 455-462
• [Journal Article] Reciprocal roles of Msx2 in regulation of osteoblast and adipocyte differentiation. (2004)
  • Author(s): Ichida F
  • Journal Title: J Biol Chem 279 Pages: 34015-34022
• [Journal Article] Critical roles of c-Jun signaling in regulation of NFAT family and RANKL-regulated osteoclast differentiation (2004)
  • Author(s): Ikeda F
  • Journal Title: J Clin Invest 114 Pages: 475-484
• [Journal Article] Differential Roles of Smad1 and p38 Kinase in the regulation of PPAR_γ during BMP2-induced Adipogenesis (2003)
  • Author(s): Hata K
  • Journal Title: Mol Biol Cell 14 Pages: 545-555
• [Journal Article] PTHrP Rescues ATDC5 Cells from Apoptosis Induced by FGF Receptor 3 Mutation (2003)
  • Author(s): Yamanaka Y
  • Journal Title: J Bone Mineral Res 18 Pages: 1395-1403
• [Journal Article] The Role of Smads in BMP Signaling. (2003)
  • Author(s): Nishimura R
  • Journal Title: Front Biosci 8
• [Journal Article] Insulin-like growth factor-1 (IGF-1) rescues the mutated FGF receptor 3 (G380R) expressing ATDC5 cells from apoptosis through phosphatidylinositol 3-kinase and mitogen-activated protein kinase (2003)
  • Author(s): Koike M
  • Journal Title: J Bone Miner Res 18 Pages: 2043-2051
• [Journal Article] Bone morphogenetic protein receptor signaling is necessary for normal murine postnatal bone formation. (2002)
  • Author(s): Zhao M
  • Journal Title: J Cell Biol. 157 Pages: 1049-1060
• [Journal Article] Cbfa1 induces osteoblastic differentiation of C2C12 cells without interactions with Smad1 and Smad5 (2002)
  • Author(s): Nishimura R
  • Journal Title: Bone 31 Pages: 303-312
• [Journal Article] A Bone-Seeking Clone Exhibits Different Biological Properties from the MDA-MB-231 Parental Human Breast Cancer Cells and a Brain-seeking Clone in Vivo and In Vitro (2001)
  • Author(s): Yoneda T
  • Journal Title: J Bone Miner Res 16 Pages: 1486-1495
• [Journal Article] Decreased c-Src expression enhances osteoblast differentiation and bone formation (2000)
  • Author(s): Marzia M
  • Journal Title: J Cell Biol 151 Pages: 311-320
• [Book] フロンティバイオデンティストリー先端歯科医学の創生 (2005)
  • Author(s): 浜田 茂幸
  • Total Pages: 273
  • Publisher: 大阪大学出版会
• [Publications] Koike M: "Insulin-like growth factor-1(IGF-1) rescues the mutated FGF receptor 3 (G380R) expressing ATDC5 cells from apoptosis through phosphatidylinositol 3-kinase and mitogen activated protein kinase"J Bone Miner Res. 18. 2043-2051 (2003)
• [Publications] Nishimura R: "The Role of Smads in BMP Signaling."Front Biosci. 8. S275-S284 (2003)
• [Publications] Yamanaka Y: "PTHrP Rescues ATDC5 Cells from Apoptosis Induced by FGF Receptor 3 Mutation"J Bone Miner Res. 18. 1395-1403 (2003)
• [Publications] Hata K: "Differential Roles of Smad1 and p38 Kinase in the regulation of PPARγ during BMP2-induced Adipogenesis"Mol Biol Cell. 14. 545-555 (2003)
• [Publications] Nishimura R: "Cbfa1 induces osteoblastic differentiation of C2C12 cells without interactions with Smad1 and Smad5"Bone. 31. 303-312 (2002)
• [Publications] Zhao M: "Bone morphogenetic proton receptor signaling is necessary for normal murine postnatal bone formation."J Cell Biol.. 157. 1049-1060 (2002)
• [Publications] Yoneda T: "A Bone-Seeking Clone Exhibits Different Biological Properties from the MDA-MB-231 Parental Human Breast Cancer Cells and a Brain-seeking Clone in Vivo and in Vitro"J Bone Miner Res. 16. 1486-1495 (2001)
• [Publications] Mazia M: "Decreased c-Src expression enhances osteoblast differentiation and bone formation"J Cell Biol. 151. 311-320 (2000)
• [Publications] Hata K: "Differential Roles of Smad1 and p38 Kinase in the regulation of PPARγ during BMP2-induced Adipogenesis"Mol Biol Cell. (印刷中). (2003)
• [Publications] Nishimura R: "Cbfa1 induces osteoblastic differentiation of C2C12 cells without interactions with Smad1 and Smad5"Bone. 31. 303-312 (2002)
• [Publications] Zhao M: "Bone morphogenetic protein receptor signaling is necessary for normal murine postnatal bone formation"J Cell Biol.. 157. 1049-1060 (2002)
• [Publications] Yoneda T: "A Bone-Seeking Clone Exhibits Different Biological Properties from the MDA-MB-231 Parental Human Breast Cancer Cells and a Brain-seekina Clone in Vivo and In Vitro"J Bone Miner Res. 16. 1486-1495 (2001)
• [Publications] Michigami T: "Cell-cell contact between marrow stromal cells and myeloma cells via VCAM-1 andα4β1 integrin enhances production of osteoclast-stimulating activity in culture"Blood. 96. 1953-1960 (2000)
• [Publications] Marzia M: "Decreased c-Src expression enhances osteoblast differentiation and bone formation"J Cell Biol. 151. 311-320 (2000)
• [Publications] Lai CF: "Transforming Growth Factor-beta Up-regulates the beta 5 Integrin Subunit Expression via Sp1 and Smad Sianaling"J Biol Chem. 275. 36400-36406 (2000)
• [Publications] Yoneda T: "Actions of bisphosphonate on bone metastasis in animal models of breast cancer"Cancer. 88. 2979-2988 (2000)
• [Publications] Ji X et al.: "Patterns of gene expression associated with BMP-2-induced osteoblast and adipocyte differentiation of mesenchymal progenitor cell 3T3-F442A"J Bone Miner Met. 18. 132-139 (2000)
• [Publications] Y Takaoka et al.: "The pancreas : A storehouse of protein anabolic factors and bone/calcium metabolism-regulating factors"J Bone Miner Met. 18. 2-8 (2000)
• [Publications] Toshiyuki Yoneda et al.: "Cellular and molecular basis of preferential metastasis of breast cancer to bone"J Orthop Sci. 5. 75-81 (2000)
• [Publications] Michigami T et al.: "Cell-cell contact between marrow stromal cells and myeloma cells via VCAM-1 and α4β1 integrin enhances production of osteoclast-stimulating activity in culture"Blood. 96. 1953-1960 (2000)
• [Publications] Marzia M et al.: "Decreased c-Src expression enhances osteoblast differentiation and bone formation"J Cell Biol. 151. 311-312 (2000)
• [Publications] Lal CF et al.: "Transforming Growth Factor-beta Up-regulates the beta 5 Integrin Subunit Expression via Sp1 and Smad Signaling"J Biol Chem. 275. 36400-364 (2000)
• [Publications] Yoneda T et al.: "Actions of bisphosphonate on bone metastasis in animal models of breast cancer"Cancer. 88. 2979-2988 (2000)
• [Publications] Yoneda T et al.: "A Bone-Seeking Clone Exhibits Different Biological Properties from the MDA-MB-231 Parental Human Breast Cancer Cells and a Brain-seeking Clone in Vivo and in Vitro"J Bone Miner Res. 16. 1486-1495 (2001)
• [Publications] Ji X et al.: "Patterns of gene expression associated with BMP-2-induced osteoblast and adipocyte differentlation of mesenchymal progenitor cell 3T3-F442A"J Bone Miner Met. 18. 132-139 (2000)
• [Publications] Y Takaoka et al.: "The pancreas : A storehouse of protein anabolic factors and bone/calcium metabolism-regulating factors."J Bone Miner Met. 18. 2-8 (2000)
• [Publications] Toshiyuki Yoneda et al.: "Cellular and molecular basis of preferential metastasis of breast cancer to bone."J Orthop Sci. 5. 75-81 (2000)
• [Publications] Michigami T et al: "Cell-cell contact between marrow stromal cells and myeloma cells via VCAM-1 andα4β1 integrin enhances production of osteoclast-stimulating activity in culture"Blood. 96. 1953-60 (2000)
• [Publications] Marzia M et al: "Decreased c-Src expression enhances osteoblast differentiation and bone formation"J Cell Biol. 151. 311-20 (2000)
• [Publications] Lai CF et al: "Transforming Growth Factor-beta Up-regulates the beta 5 Integrin Subunit Expression via Sp1 and Smad Signaling."J Biol Chem. 275. 36400-36406 (2000)
• [Publications] Yoneda T et al: "Actions of bisphosphonate on bone metastasis in animal models of breast cancer."Cancer. 88. 2979-2988 (2000)
• [Publications] Yoneda T et al: "A Bone-Seeking Clone Exhibits Different Biological Properties from the MDA-MB-231 Parental Human Breast Cancer Cells and a Brain-seeking Clone in Vivo and in Vitro"J Bone Miner Res. (印刷中). (2001)