転写因子Cbfalを中心とした骨格形成機構の解明

• Project/Area Number: 12137204
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Nagasaki University (2004); Osaka University (2000-2003)
• Principal Investigator: 小守 壽文 長崎大学, 大学院・医歯薬学総合研究科, 教授 (00252677)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥53,500,000 (Direct Cost: ¥53,500,000); Fiscal Year 2004: ¥13,400,000 (Direct Cost: ¥13,400,000); Fiscal Year 2003: ¥13,600,000 (Direct Cost: ¥13,600,000); Fiscal Year 2002: ¥13,300,000 (Direct Cost: ¥13,300,000); Fiscal Year 2001: ¥13,200,000 (Direct Cost: ¥13,200,000)
• Keywords: Cbfa1 / Runx2 / Runx3 / 軟骨細胞 / Ihh / ランクスファミリー / 骨格形成 / 転写因子 / 破骨細胞分化 / RANKL / OPG / 脂肪細胞分化 / IL-11 / 骨芽細胞 / 骨 / 軟骨 / トランスジェニックマウス / ノックアウトマウス / 血管進入 / 間葉系細胞 / 骨形成

Research Abstract

大部分の骨は、内軟骨生骨化によって形成される。すなわち、軟骨細胞は成熟後、骨に置換される。転写因子Cbfa1/Runx2のノックアウトマウスは、軟骨で骨格は形成されるが、骨形成を全く見ない。これは、骨芽細胞の分化が起こらないことから生じるが、その軟骨細胞の成熟も阻害されている。実際、Runx2およびドミナントネガティブ型Runx2の軟骨細胞特異的トランスジェニックマウスでは、軟骨細胞の分化促進および抑制がそれぞれ見られた。したがって、Runx2は、軟骨細胞の成熟にも重要な因子である。しかし、Runx2のノックアウトマウスの一部の骨格では軟骨細胞の成熟が見られ、Runx2以外の因子も軟骨細胞の成熟に関与していると考えられる。そこで、同じランクスファミリーに属し、軟骨細胞に発現しているCbfa3/Runx3のヘテロ変異マウスとRunx2ヘテロ変異マウスを交配し、最終的にRunx2とRunx3のダブルノックアウトマウスを作製した。このダブルノックアウトマウスでは、全身の骨格で、軟骨細胞の成熟が完全に抑制されていた。したがって、Runx2とRunx3が、軟骨細胞の成熟に必須な因子であることが明らかとなった。さらにRunx2とRunx3の遺伝子量の減少に比例して、四肢の著明な短縮が認められた。そして、軟骨細胞の増殖に重要なインディアンヘッジホグ(Ihh)の発現が消失していた。Runx2ノックアウトマウスでは、軟骨細胞の増殖が低下しており、Runx2は、Ihhのプローモーターに直接結合し、その転写を促進していた。一方、Runx3は間接的にIhhの発現を誘導していると考えられた。したがって、Runx2とRunx3は、Ihhの発現調節を介して、軟骨細胞の増殖も調節していることが明らかとなった。

Research Products

• [Journal Article] Dexamethasone inhibits insulin-induced chondrogenesis of ATDC5 cells by preventing PI3K-Akt signaling and DNA binding of Runx2. (2004)
  • Author(s): T.Fujita
  • Journal Title: J Cell.Biochem. 93 Pages: 374-383
• [Journal Article] Runx2 induces osteoblast and chondrocyte differentiation and enhances their migration by coupling with PI3K-Akt signaling. (2004)
  • Author(s): T.Fujita
  • Journal Title: J Cell Biol. 166 Pages: 85-95
• [Journal Article] Statins inhibit osteoblast migration by inhibiting Rac-Akt signaling. (2004)
  • Author(s): R.Fukuyama
  • Journal Title: Biochem.Biophys.Res.Commun. 315 Pages: 636-642
• [Journal Article] Menin Is Required for BMP2-induced osteoblastic differentiation through the interaction with Smad1/5 and Runx2. (2004)
  • Author(s): H.Sowa
  • Journal Title: J Biol.Chem. 279 Pages: 40267-40275
• [Journal Article] Aged mice require full transcription factor, Runx2/Cbfa1, gene dosage for cancellous bone regeneration after bone marrow ablation. (2004)
  • Author(s): K.Tsuji
  • Journal Title: J.Bone Miner.Res. 19 Pages: 1481-1489
• [Journal Article] Msx2 regulates osteoblast differentiation in Cbfa1/Runx2-independent fashion and inhibits adipocyte differentiation by suppressing PPARg and C/EBP family. (2004)
  • Author(s): F.Ichida
  • Journal Title: J Biol.Chem. 279 Pages: 34015-34022
• [Journal Article] Runx2-deficient mice lack mandibular condylar cartilage and have deformed Meckel's cartilage. (2004)
  • Author(s): S.Shibata
  • Journal Title: Anat.Embryol. 208 Pages: 273-280
• [Journal Article] Expression of Dentin Matrix Protein 1 (DMP1) during Fracture Healing. (2004)
  • Author(s): S.Toyosawa
  • Journal Title: Bone 35 Pages: 553-561
• [Journal Article] Runx2 and Runx3 are essential for chondrocyte maturation and Runx2 regulates limb growth through induction of Indian hedgehog. (2004)
  • Author(s): C.A.Yoshida
  • Journal Title: Genes Dev. 18 Pages: 952-963
• [Journal Article] Micro-CT evaluation of tooth, calvaria and mechanical stress-induced tooth movement in adult Runx2/Cbfa1 heterozygous knock-out mice. (2004)
  • Author(s): CR.Chung
  • Journal Title: J.Med.Dent.Sci. 51 Pages: 105-113
• [Journal Article] Delayed tooth eruption and suppressed osteoclast number in the eruption pathway of heterozygous Runx2/Cbfa1 knockout mice. (2004)
  • Author(s): S.Yoda
  • Journal Title: Arch.Oral Biol. 49 Pages: 435-442
• [Journal Article] Isolation and characterization of a mesenchymal cell line that differentiates into osteoblasts in response to BMP-2 from calvariae of GFP transgenic mice (2004)
  • Author(s): A.Kadowaki
  • Journal Title: Bone 34 Pages: 993-1003
• [Journal Article] Impairment of bone healing by insulin receptor substrate-1 deficiency. (2004)
  • Author(s): T.Shimoaka
  • Journal Title: J.Biol.Chem. 279 Pages: 15314-15322
• [Journal Article] Evaluation of 9.4-T MR microimaging in assessing normal and defective fetal bone development : comparison of MR imaging and histological findings. (2004)
  • Author(s): Y.Ichikawa
  • Journal Title: Bone 34 Pages: 619-628
• [Journal Article] Expression of dentin matrix protein 1 in tumors causing oncogenic osteomalacia. (2004)
  • Author(s): S.Toyosawa
  • Journal Title: Modern Pathol. 17 Pages: 573-578
• [Journal Article] mRNA expression and protein localization of dentin matrix protein 1 during dental root formation. (2004)
  • Author(s): S.Toyosawa
  • Journal Title: Bone 34 Pages: 124-133
• [Journal Article] Runx2 deficiency in chondrocytes causes adipogenic changes in vitro. (2004)
  • Author(s): H.Enomoto
  • Journal Title: J.Cell Sci. 117 Pages: 417-425
• [Journal Article] Regulation of skeletal development by the Runx family of transcription factors.
  • Author(s): T.Komori
  • Journal Title: J.Cell.Biochem. (in press)
• [Journal Article] Inhibition of Cdk6 expression through p38 MAP kinase is involved in differentiation of mouse prechondrocyte ATDC5.
  • Author(s): T.Moro
  • Journal Title: J.Cell.Physiol. (in press)
• [Publications] A.Kadowaki: "Isolation and characterization of a mesenchymal cell line that differentiates into osteoblasts in response to BMP-2 from calvariae of GFP transgenic mice."Bone. (in press).
• [Publications] T.Shimoaka: "Impairment of bone healing by insulin receptor substrate-1 deficiency."J.Biol.Chem.. (in press).
• [Publications] Y.Ichikawa: "Evaluation of 9.4-T MR micro-imaging in assessing normal and defective fetal bone development : comparison of MR imaging and histological findings."Bone. (in press).
• [Publications] S.Toyosawa: "Expression of dentin matrix protein 1 in tumors causing oncogenic osteomalacia."Modern Pathol.. (in press).
• [Publications] R.Fukuyama: "Statins inhibit osteoblast migration by inhibiting Rac-Akt signaling."Biochem.Biophys.Res.Commun.. 315. 636-642 (2004)
• [Publications] S.Toyosawa: "mRNA expression and protein localization of dentin matrix protein 1 during dental root formation."Bone. 34. 124-133 (2004)
• [Publications] H.Enomoto: "Runx2 deficiency in chondrocytes causes adipogenic changes in vitro."J.Cell Sci.. 117. 417-425 (2004)
• [Publications] J.Pratap: "Cell growth regulatory role of Runx2 during proliferative expansion of preosteoblasts."Cancer Res.. 63. 5357-5362 (2003)
• [Publications] H.Enomoto: "Induction of osteoclast differentiation by Runx2 through RANKL and OPG regulation and partial rescue of osteoclastogenesis in Runx2-/- mice by RANKL transgene."J.Biol.Chem.. 278. 23971-23977 (2004)
• [Publications] M.Takamoto: "Hedgehog signaling enhances core-binding factor a1 and receptor activator of nuclear factor-kappaB ligand (RANKL) gene expression in chondrocytes."J.Endocrinol.. 177. 413-421 (2003)
• [Publications] K.Hirata: "Transplantation of skin fibroblasts expressing BMP-2 promotes bone repair more effectively than those expressing Runx2."Bone. 32. 502-512 (2003)
• [Publications] T.Komori: "Requisite roles of Runx2 and Cbfb in skeletal development."J.Bone Miner.Metab.. 21. 193-197 (2003)
• [Publications] K.Hirata: "Transplantation of skin fibroblasts expressing BMP-2 promotes bone repair more effectively than those expressing Runx2"Bone. (in press).
• [Publications] T.Komori: "Requisite roles of Runx2 and Cbfb in skeletal development"J.Bone Miner.Metab.. (in press).
• [Publications] M.Iwamoto: "Runx2 expression and action in chondrocytes are regulated by retinoid signaling and parathyroid hormone-related peptide(PTHrP)"Osteoarthritis and Cartilage. 11. 6-15 (2003)
• [Publications] C.A.Yoshida: "Core-binding factor β interacts with Runx2 and is required for skeletal development"Nature Genetics. 32. 633-638 (2002)
• [Publications] I.Taniuchi: "Differential requirements for Runx proteins in CD4 repression and epigenetic silencing during T lymphocyte development"Cell. 111. 621-633 (2002)
• [Publications] M.Enomoto-Iwamoto: "The Wnt antagonist Frzb-1 regulates chondrocyte maturation and long bone development during limb skeletogenesis"Dev.Biol.. 251. 142-156 (2002)
• [Publications] T.Komori: "Runx2, a multifunctional transcription factor in skeletal development"J.Cell.Biochem.. 87. 1-8 (2002)
• [Publications] Z.Shen: "Negative regulation of BMP/Smad signaling by Cas-interacting zinc finger protein in osteoblasts"J.Biol.Chem.. 277. 29840-29846 (2002)
• [Publications] T.Komori: "The Growth Plate"I.M.Shapiro, B.Boyan, H.C.Anderson ed. (IOS Press, Amsterdam). 19-23 (2002)
• [Publications] M.Himeno: "Impaired vascular invasion of Cbfa1-deficient cartilage engrafted in spleen"J.Bone Miner.Res.. (in press).
• [Publications] M.J.G.Jimenez: "A regulatory cascade involving retinoic acid, Cbfa1, and matrix metalloproteinases is coupled to the development of a process of perichondrial invasion and osteogenic differentiation during bone formation"J.Cell Biol.. 155. 1333-1344 (2001)
• [Publications] S.Toyosawa: "Dentin matrix protein is predominantly expressed in chicken and rat osteocytes, but not in osteoblasts"J.Bone Miner.Res.. 16. 2017-2026 (2001)
• [Publications] W.Liu: "Overexpression of Cbfa1 in osteoblasts inhibits osteoblast maturation and causes osteopenia with multiple fractures"J.Cell Biol.. 155. 157-166 (2001)
• [Publications] M.Enomoto-Iwamoto: "Participation of Cbfa1 in regulation of chondrocyte maturation"Osteoarthritis Cartilage. 9. S76-S84 (2001)
• [Publications] C.Ueta: "Skeletal malformations caused by overexpression of Cbfa1 or its dominant negative form in chondrocytes"J.Cell Biol.. 153. 87-100 (2001)
• [Publications] T.Komori: "Cbfa1, a key transcription factor for skeletogenesis"In Recent Research Development in Molecular & Cellular Biology, S. G. Pandalai ed. (Research Signpost). 13 (2001)
• [Publications] Ueta,C.,: "Skeletal malformations caused by overexpression of Cbfa1 or its dominant negative form in chondrocytes."J.Cell Biol.. (in press).
• [Publications] Ikegame,M.,: "Tensile stress induces bone morphogenetic protein 4 (BMP-4) in preosteoblastic and fibroblastic cells, which later differentiate into osteoblasts leading to osteogenesis in the mouse calvariae in organ culture."J.Bone Miner.Res.. 16. 24-32 (2001)
• [Publications] Lee,K.,: "Runx2/PEBP2αA/Cbfa1 is a common target of TGF-b1 and BMP-2 and cooperation between Runx2 and Smad5 induces osteoblast specific gene expression in the pluripotent mesenchymal precursor cell line C2C12."Mol.Cell.Biol.. 20. 8783-8792 (2000)
• [Publications] Komori,T.: "A fundamental transcription factor for bone and cartilage."Biochem.Biophys.Res.Commun.. 276. 813-816 (2000)
• [Publications] Yamaguchi,A.,: "Regulation of osteoblast differentiation mediated by bone morphogenetic proteins, bedgehogs, and Cbfa1."Endocrine Rev.. 21. 393-411 (2000)
• [Publications] Kobayashi,H.,: "Multilineage differentiation of Cbfa1-deficient calvarial cells in vitro."Biochem.Biophys.Res.Commun.. 273. 630-636 (2000)
• [Publications] Komori,T.: "Cbfa1, In Recent Research Development in Molecular & Cellular Biology"S.G.Pandalai ed.(Research Signpost)(in press.).