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Search for key molecules in bone and cartilage differentiation

Research Project

Project/Area Number 13557153
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Functional basic dentistry
Research InstitutionTokyo Medical and Dental University

Principal Investigator

NIFUJI Akira  Tokyo Medical and Dental University, Medical Research Institute, Department of Molecular Pharmacology, Associate Professor, 難治疾患研究所, 助教授 (00240747)

Co-Investigator(Kenkyū-buntansha) NODA Masaki  Tokyo Medical and Dental University, Medical Research Institute, Department of Molecular Pharmacology, Professor, 難治疾患研究所, 教授 (50231725)
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥12,000,000 (Direct Cost: ¥12,000,000)
Fiscal Year 2003: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2002: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2001: ¥5,600,000 (Direct Cost: ¥5,600,000)
KeywordsDifferentiation / Bone / cloning / Cartilage / BMP
Research Abstract

Various type of signaling molecules and transcription factors are involved in bone and cartilage differentiation. Studies based on loss of function indicate that Runx 2, Osterix, and Sox 9 are indispensable for development of such hard tissues. Although those molecules are critical for skeletetogenesis, there still would be other uncharacterized molecules that govern bone and cartilage differentiation. To search for such molecules, we tried to establish a functional screening assay system specifically utilized for cartilage differentiation. We used type Xl collagen promoter as a monitor to differentiate into chondrocyte, since endogenous type Xl collagen expression is induced following the cartilage differentiation. We transfected collagen promoter plus beta-galactosidase construct (pXlcol-Z) into a teratocarcinoma derived fibroblastic cell line Fl 2. Using antibiotics we isolated stably expressed transformants. Among them, we selected one transformant, which barely expressed LacZ at steady state level but is induced to express upon treatment with BMP. Since it is known that BMP induced chondrogenic differentiation in vitro, BMP responsive promoter may reflect at least one aspect of chondrogenic differentiation. We named such transformants as pR5-1 cells. We next analyzed Lac~ Z expression by FACS. BMP treated pR5-l cells showed higher amount of LacZ positive cells than untreated cells, showing that FACS can be useful to collect cells in which the promoter is activated. We then prepared chondrocyte specific cDNAs. We used teratocarcinoma derived mesodermal cell line Cl as a source of RNA. We prepared cDNA libraries based on retrovirus vector plasmid and transfected them into a packaging cell line. Then we collected supernatant of those cells. We infected retrovirus cDNA library into pR5-1 cells. We, then, collected highly LacZ positive pR5-1 cells. We are now under way to isolate and characterize integrated genes.

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Nifuji A, Miura N, Kato N, Kellermann O, Noda M: "Bone morphogenetic protein regulation of forkhead/winged helix transcription factor Foxc2 (Mfh1) in a murine mesodermal cell line C1 and in skeletal precursor cells."J Bone Miner Res.. 16・10. 1765-1771 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Asou Y, Nifuji A, Tsuji K, Shinomiya K, Olson EN, Koopman P, Noda M: "Coordinated expression of scleraxis and Sox9 genes during embryonic development of tendors and cartilage."J Orthop Res.. 20・4. 827-833 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shen ZJ, Nakamoto T, Tsuji K, Nifuji A, Miyaznono K, Komori T, Hirai H: "Negative regulation of bone morphogenetic protein/Smad signaling by Cas-interacting zinc finger protein in osteoblasts."J Biol Chem.. 277・33. 29840-29846 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Takamoto M, Tsuji K, Yamashita T, Sasaki H, Yano T, Taketani Y, Komori T, Nifuji A, Noda M: "Hedgehog signaling enhances core-binding factor a1 and receptor activator of nuclear factor kappa-β ligand (RANKL) gene expression in chondrocytes."Journal of Endocrinology. 177. 413-421 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nifuji A, Kellermann O, Noda M: "Noggin inhibits chondrogenic but not osteogenic differentiation in mesodermal stem cell line C1 and skeletal cells."Endocrinology. 145・7. 3434-3442 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nifuji A, Miura N, Kato N, Kellermann O, Noda M: "Bone morphogenetic protein regulation of forkhead/winged helix transcription factor Foxc2 (MTh 1) in a murine mesoderrnal cell line Cl and in skeletal piecursor cells."J Bone Min Res. 16(10). 1765-1771 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Asou Y, Nifuji A.Tsuji K, Shinomiya K, Olson EN, Koopman P, Noda M.: "Coordinated expression of scieraxis and Sox9 genes during embryonic development of tendons and cartilage."J Orthop Res. 20(4). 827-833 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shen ZJ, Nakamoto T, Tsuji K, Nifuji A, Miyazono K, Komon T, Hirai H, Noda M.: "Negative regulation of bone morphogenetic protein/Smad signaling by Cas-interacting zinc finger protein in osteoblasts."J Biol Chem.. 277(33). 29840-28946 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Takamoto M, Tsuji K, Yamashita T, Sasaki H, Yano T, Taketani Y, Komon T.Nifuji A.Noda M: "Hedgehog signaling enhances core-binding factor al and receptor activator of nuclear factor-lcappaB ligand (RANKL) gene expression in chondrocytes"Journal of Endocrinology. 177. 413-421 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nifuji A.Kellermann O, Noda M.: "Noggin inhibits chondrogenic but not osteogenic differentiation in mesodermal stem cell line cl and skeletal cells."Endocrinology. 145. 3434-3442 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yagi K, Tsuji K, Nifuji A, et al.: "Bone morphogenetic protein-2 enhances osterix gene expression in chondrocytes"J Cell Biochem. 88・6. 1077-1083 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Salingcarnboriboon R, Yoshitake H, et al.: "Establishment of tendon-derived cell lines exhibiting pluripotent mesenchymal stem cell-like property"Exp Cell Res. 287・2. 289-300 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Asou Y, Nifuji A, Tsuji K, Shinomiya K, Olson EN, Koopman P, Noda M.: "Coordinated expression of scieraxis and Sox9 genes during embryonic development of tendons and cartilage"J Orthop Res.. 20(4). 827-833 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Shen ZJ., Nakamoto T., Tsuji K., Nifuji A., Miyazono K., Komori T., Hirai H., Noda M.: "Negative regulation of bone morphogenetic protein/Smad signaling by Cas-interacting zinc finger protein in osteoblasts"J Biol Chem.. 277(33). (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nifuji A, Miura N, Kato N, Kellermann O, Noda M.: "Bone morphogenetic protein regulation of forkhcad/winged helix transcription factor Foxc2 (Mfhl) in a murine mesodermal cell line C1 and in skeletal precursor cells"J Bone Miner Res. 16・10. 1765-1771 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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