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Basic research regarding gene therapy for leukemia by using homologous recombination

Research Project

Project/Area Number 08457275
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Hematology
Research InstitutionNiigata University

Principal Investigator

TAKAHASHI Masuhiro  Niigata University, College of Biomedical Technology, Department of Medical Technology, Professor, 医療技術短期大学部, 教授 (90179531)

Co-Investigator(Kenkyū-buntansha) FURUKAWA Tatsuo  Niigata University, School of Medicine, Division of Bioclean Room, Associate Pro, 医学部・附属病院, 助教授 (00272849)
KOIKE Tadashi  Niigata University, School of Medicine, Division of Blood Transfusion, Associate, 医学部・附属病院, 助教授 (30170161)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1997: ¥700,000 (Direct Cost: ¥700,000)
Keywordshomologous recombination / bcr-abl / thymidine kinase / neo / gene replacement / bvr-abl融合遺伝子 / homologous recombination / leukemia / gene therapy / BCR-ABL oncogane / RT-PCR / Souther Blotting / Neo / Thymidine kinase
Research Abstract

Ideal somatic gene therapy for genetic diseases is to treat by the replacement of an abnormal gene with a normal gene and to normalize the characteristics of abnormal cells. Up to now, repair of an abnormal gene can be performed only by homologous recombination between that abnormal gene and the gene for correction. Our study was designed to discover whether homologous recombination occurs in hematopoietic cells and if is possible to establish gene therapy using homologous recombination. Murine hematopoietic cell line, FDC-P2, which was factor-dependent but became factor-independent by being transfected with p210bcr-abl was used for the target cell for homologous recombination. The plasmid for homologous recombination was made by inserting neo-gene in the region of bcr-abl, thymidine kinase-gene in the region outside of bcr-abl, and placing bcr-abl region reversely for defining the homologous recombination region to bcr-abl. Transfection was done by electroporation and selection was performed by adding G418 and gancyclovir. One clone was established and confirmed to be replaced by the plasmid made for homologous recombination by RT-PCR findings of thymidine kinase (-), neo (+), bcr-abl (-) and a reasonable change of thr band by Southern blot analysis. The clone became factor-independent. These findings suggest the possibility of homologous recombination-associated gene replacement therapy for hematological diseases including leukemia.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Takahashi M: "Effects of transfection of p210 bcr-abl and bcr-v-abl into the factor-dependent human leakemia cell line HSM-911." Leuk Res. 21(11/12). 1115-1123 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Toba K: "Characterization and semiticrty to IL-2 and IFN-α of leukemer cells from apatient with lenge granular lymyhocytic leakemia associatid・・・" Leuk Res. 21(10). 941-950 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tsukada N: "The heterogenous expression of CD80. CD86 and other adhesion molecules on leukemia and lynphoma cells and their induction by IF" J Exp Clin Cancer Res. 16. 171-176 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ichimura T: "The 14-3-3 protein pinds its target proteins with a commonsite located towards the C-teanines" FEBS Lett. 413. 273-276 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Komata T: "B7-1(CD80)-transfected human glioma cells and IL-12 directly stimulate allogeneic CD8+Tells" J Immunotherapy. 20. 256-264 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takahashi M: "Confiscation of antonomous proliferation from P210 bcr-abl transfected cells by re-transfection with DNA for homologous・・・" Tranogenics. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takahashi M,Shigeno N,Takahashi H,Suzuki N,Masuko M,Nikkuni K,Toba K,Furukawa T,Aoki S,Kishi K,Koike T,Aizawa Y: "Effects of transfection of P210bcr-abl and bcr-v-abl into the factor-dependent human leukemia cell line HSM-911" Leuk Res. 21 (11/12). 1115-1123 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Toba K,Koike T,Takahashi M,Kishi K,Hashimoto S,Takahashi H,Uesugi Y,Ishikawa T,Aoki S,Maeo S,Naito M,Aizawa Y,Shibata A: "Characterization and sensitivity to interleukin-2 and interferon-alpha of leukemic cells from a patient with large granular lymphocytic leukemia associated with chronic active Epstein-Barr virus infection" Leuk Res. 21 (10). 941-950 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tsukada N,Aoki S,Maruyama S,Kishi K,Takahashi M,Aizawa Y: "The heterogeneous expression of CD80, CD86 and other adhesion molecules on leukemia and lymphoma cells and their induction by interferon" J Exp Clin Cancer Res. 16. 171-176 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ichimura T,Ito N,Itagaki C,Takahashi M,Horigome T,Omata S,Ohno S,Isobe T: "The 14-3-3 protein binds its target proteins with a common site located towards the C-terminus." FEBS Lett. 413. 273-276 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Komata T,Tanaka R,Yamamoto K,Oda T,Ono K,Yoshida S,Takahashi M: "B7-1 (CD80) -transfected human glioma cells and interleukin-12 directly stimulate allogeneic CD8+ T cells" J Im munotherapy. 20. 256-264 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takahashi M,Shigeno N,Suzuki N,Takahashi H,Nikkuni K,Masuko M,Sakaue M,Furukawa T,Kishi K,Koike T,Aizawa Y: "Confiscation of Autonomous Proliferation from P210bcr-abl Transfected Cells by Re-transfection with DNA for Homologous Recombination" Transgenics. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takahashi M,Narita M,Shigeno N,Takahashi H,Furukawa T,Koike T,Aizawa Y: "Cotinine (metabolite of nicotine) suppresses the growth of hematopoietic progenitor cells at the concentration range equivalent to its serum concentration in smokers" Stem Cells. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takahashi M: "Effects of transgection of P210 bcr - abl and bcr - v - abl into the factor depandent fuman leakemia cell line HSM - 911" Leuk Res. 21(11/12). 1115-1123 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Toba K: "Chanactecigation and semectivity to IL-2 and IFN - α of leukement cells from apatient uithe lange granula lymphocytic leupcmua assouiated" Leuk Res. 21(10). 941-950 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Tsukada N: "The heterogenous expreasian of CD86 and other adhession moleeules on leukemia and lympcoma cells and thein imduction by IF" J Exp Clin Canca Res. 16. 171-176 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ichimura T: "The 14 - 3 - 3 protein kinds its tanget proteins with a commor site located towards the cteancino" FEBS Lett. 413. 273-276 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Komata T: "B7 1 (CD80) - tranfected human gluoma cells and IL - 12 directly stimu late allagenaic CD8 + T cells" J lmmunotherapy. 20. 256-264 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Takahashi M: "Confiscatron of autonamaus proliferation from P210 bcr - abl transfecterd cells by retransfetion with DNA for fomologous" Transgenics. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Tobak. et al: "Profile of cell cycle in hematopoietic malignancy by DNAIRNA quantitation using 7AADIPY" Experimental Hematology. 24(8). 894-901 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hanawa H. et al: "Novel analysis of minimal rasidual disease in leukemia with TCR beta rearrangement" Leukemia Research. (in press).

    • Related Report
      1996 Annual Research Report
  • [Publications] 鈴木訓充: "相同組換えによる異常遺伝子を標的とした遺伝子破壊の試み-増血器腫瘍への遺伝子治療の可能性" 新潟医学会雑誌. (in press).

    • Related Report
      1996 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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