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Enzyme Replacement Therapy for Fabry Disease :Production of Efficient α-Galactosidase

Research Project

Project/Area Number 13672409
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用薬理学・医療系薬学
Research InstitutionTokyo Metropolitan Organization for Medical Research

Principal Investigator

KASE Ryoichi  The Tokyo Metropolitan Institute of Medical Science, 東京都臨床医学総合研究所, 研究員 (20150203)

Co-Investigator(Kenkyū-buntansha) SIMMOTO Michie  The Tokyo Metropolitan Institute of Medical Science, 東京都臨床医学総合研究所, 研究員 (20216237)
SAKURABA Hitoshi  The Tokyo Metropolitan Institute of Medical Science, 東京都臨床医学総合研究所, 研究員 (60114493)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordsFadry Disease / α-Galactosidase / Lysosome / Enzyme Replacement / α-ガラクトシターゼ
Research Abstract

The genetic defect of α-galactosidase encoded by a gene localized to the X- chromosomal region, Xq22, results in Fabry disease. This disease is characterized by the systemic intralysosomal accumulation of neutral sphingolipids, predominantly globotriaosylceramide. In our basic studies of α-galactosidase enzyme replacement therapy, first, we constructed the clones of the methylotropic yeast Pichia pastois. Then, the glyco-chains of the recombinant α-galactosidase was trimmed by α-mannosidase, and the effective up-take by Fabry fibroblasts was observed. The administrated α-galactosidase could be recaptured via the mannose 6-phosphate receptors by cells, a phenomenon commonly referred as receptor-mediated endocytosis. Second, we introduced site-directed mutations into α-galactosidase. X-ray crysrarographic analysis revealed that two Asp (170 and 231) residues were involved in catalitic-site of the enzyme. We made Asp (D) to Glu (E) changed mutants D170E, D231E, and D170E/D231E (double mutation), respectively. These mutants were Vmax mutants. The kinetic studies revealed that D231 played the important role in the rate limiting step of the α-galactosidase reaction

Report

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

    (17 results)

All Other

All Publications (17 results)

  • [Publications] utsumi K.: "Western blotting analysis of the -hexosaminidase -and -subunits in cultured fibroblasts from cases of various forms of GM2 gangliosidosis"Acta Neurol.Scand.. 105. 427-430 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Naganawa Y.: "In Vitro Study of Encapsulation Therapy for Fabry Disease Using Genetically Engineered CHO Cell Line"Cell Transplant. 11. 325-329 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Takahashi H.: "Long-term systemic therapy of Fabry disease adeno-associated virus-mediated muscle-directed gene transfer"Proc.Natl.Acad.Sci.USA. 99. 13777-13782 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Chiba Y.: "Production in yeast of alpha-galactosidase A, a lysosomal enzyme applicable to enzyme replacement therapy for Fabry disease"Glycobiology. 12. 821-828 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okumiya T.: "Imbalanced substrate specificity of mutant β-galactosidase in patients with Morquio B disease"Mol.Genet.Metab.. 78. 51-58 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shimada T: "Long Term Correction of Lipid Storage in Multiple Organs of Fabry Mice by Direct Injection of AAV Vectors into Skeltal Muscle"Am. J. Hum. Genet.. 69. 678 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Utsumi K: "Western Blotting Analysis of the β- Hexoseaminidase α- and β-Subunits in Cultured Fibroblasts from Cases of Various Forms of GM2 Gangliosidosis"Acta Neurol. Scand.. 105. 427-430 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Naganawa Y: "In Vitro Study of Encapsulation Therapy for Fabry Disease Using Genetically Engineered CHO Cell Line"Cell Transplant.. 11. 325-329 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Takahashi H: "Long-Term Systemic Therapy of Fabry Disease by Adeno-Associated Virus-Mediated Muscle-Directed Gene Transfer"Proc. Natl. Acad. Sci. USA. 99. 13777-13782 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Chiba Y: "Production in Yeast of Alpha-Galactosidase A, a Lysosomal Enzyme Applicable to Enzyme Replacement Therapy for Fabry Disease"Glycobiology. 12. 821-828 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okumiya T: "Imbalanced Substrate Specificity of Mutant β-Galactosidase in Patients with Morquio B Disease"Mol. Genet. Metab.. 78. 51-58 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Utsumi K.: "Western blotting analysis of the β-hexosaminidase α-and β-subunits in cultured fibroblasts from cases of various forms of GM_2 gangliosidosis"Acta Neurol. Scand.. 105. 427-430 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Naganawa Y.: "In vitro study of encapsulation Therapy for Fabry disease using genetically engineered CHO cell line"Cell Transplant. 11. 325-329 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Takahashi H.: "Long-term systemic therapy of Fabry disease by adeno-associated virus-mediated muscle-directed gene transfer"Proc. Natl. Acad. Sci. USA. 99. 13777-13782 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Chiba Y.: "Production in yeast of alpha-galactosidase A, a lysosomal enzyme applicable to enzyme replacement therapy for Fabry disease"Glycobiology. 12. 821-828 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Okumiya T.: "Imbalanced substrate specificity of mutant β-galactosidase in patients with Morquio B disease"Mol. Genet. Metabol.. 78. 51-58 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Shimada T.: "Long term correction of storage in multipleorgans of Fabry mice by direct of injection AAV vectors into skeltal muscle"Am. J. Hum. Genat.. 69. 678 (2001)

    • Related Report
      2001 Annual Research Report

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

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