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New vector system for the gene therapy of abdominal cancers.

Research Project

Project/Area Number 09557103
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Digestive surgery
Research InstitutionNagoya University

Principal Investigator

NIMURA Yuji  Nagoya University School of Medicine Professor, 医学部, 教授 (80126888)

Co-Investigator(Kenkyū-buntansha) KANAI Michio  Kasugai City Hospital Chief of the Surgical Department, 助手・外科部長 (50242871)
NAGINO Masato  Nagoya University School of Medicine Associate Professor, 医学部, 講師 (20237564)
HAGIWARA Masatoshi  Tokyo Medical and Dental University School of Medicine, Professor, 難治疾患研究所, 教授 (10208423)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥12,800,000 (Direct Cost: ¥12,800,000)
Fiscal Year 1998: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1997: ¥8,800,000 (Direct Cost: ¥8,800,000)
Keywordsgene therapy / retroviral vector / peritoneal dessemination / transcriptional regulation / 腹膜播種癌 / レトロウイルスベクター / ドキシサイクリン / on / off制御 / GFP
Research Abstract

Tight control of gene expression in temporal and quantitativemanner is an invaluable tool in both basic and clinical biology. The desired characteristics of the inducible system are as follows. (I) The regulator should be activated only upon administration of an exogenousmediator, and terminated when the exogenous stimulus is removed. (ii) The mediator should be non toxic and active upon oral administration. (iii) The system should not activate other endogenous cellular genes. We constricted a retroviral vector that has an autoregulatory cassette and express the gene of interest in response to oral administration of doxycycline (dox) invivo. The cassette contains the all components of the reversetetracycline-regulated (rtTA) system (Gossen M.et al. 1995), a drugselectable marker (BSD) with the internal ribosome entry site (IRES) and the gene of interest (GFP). The retroviral long term repeat enhancer and promoter elements driv-expression of the taransactivator (rTetR-VP16) and BSD, but … More the translation was terminated at the 3'-site of BSD. The expression of GFP was controlled under the tandem tet operator sequencesand the cytomegalovirus minimal promoter in a dose-dependent manner to dox. FACS analyses showed that GFP-fluoresence was induced in two-ordermagnitude in the retrovirus-infected 208F cells dependent on the amount of dox. Furthermore oral administration of dox could induce GFP protein in thetransplanted 208F cells in the peritoneal cavity of a nude mouse. Thus, this reverse tetracycline-regulated retroviral vector (RTRRV) system allowseasy delivery of controllable genes to cultured cells and transgenicanimals. However, the control system did not work in several human cancercells. This may be attributed to the species-dependent susceptibility toretorviruses. And we found that the introduction of a toxic gene into RTRRVkilled the PA317 retrovirus packaging cells, suggesting that RTRRV still suffer from problems of higher basal levels of gene expression undernoninduced condition and pleiotropic effects on host cell genes. If we will solve these problems mentions above, this inducible retroviral vector system may pave the way to the controlled gene expression during a particular window of time in gene therapy applications. Less

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Moriguchi T,et al.: "Purification and identification of a major activator for p38 from osmotically shocked cells.Activation of MAPKK6 by osmotic shock,tumor necrosis factor-a and H202"J Biol Chem. 271. 26981-26988 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Watanabe Y,et al.: "Characterization of a serum response factor-like protein in saccharomyces cerevisiae,Rlml,which has transcriptional activity regulated by the Mpk1(Slt2)mitogen-activated protein kinase pathway"Mol Cell Biol. 17. 2615-2623 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kamimoto T,et al.: "Dymple,a novel dynamin-like high molecular weight GTPase lacking a proline-rich C-terminal domani in mammalian cells"J Biol Chem. 273. 1044-1051 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shimomura A,et al.: "Dominant negative ATF1 blocks cyclic AMP-induced neurite outgrowth in PC12D cells"J Neurochem. 70. 1029-1034 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nimura Y,et al.: "Aggressive surgical treatment of hilar cholangiocarcinoma"J Hep Bil Pancr Surg. 5. 52-61 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sakamoto E,et al.: "The pattern of infiltration at the proximal border of hilar bile duct carcinoma A histologic analysis of 62 resected cases"Ann Surg. 227. 405-411 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Moriguchi T: "Purification and identification of a major activator for p38 from osmotically shocked cells. Activation of MAPKK6 by osmotic shock, tumor necrosis factor-a and H202"J Biol Chem. 271. 26981-26988 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Watanabe Y: "Characterization of a serum response factor-like protein in saccharomyces cerevisiae, Rlm1, which has transcriptional activity regulated by the Mpk 1 (Slt 2) mitogen-activated protein kinase pathway"Mol Cell Biol. 17. 2615-2623 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kamimoto T: "Dymple, a novel dynamin-like high molecular weight GTPase lacking a proline-rich C-terminal domani in mammalian cells"J Biol Chem. 273. 1044-1051 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shimomura A: "Dominant negative ATF 1 blocks cyclic AMP-induced neurite outgrowth in PC12D cells"J Neurochem. 70. 1029-1034 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nimura Y: "Aggressive surgical treatment of hilar cholangiocarcinoma"J Hep Bil Pancr Surg. 5. 52-61 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sakamoto E: "The pattern of infiltration at the proximal border of hilar bile duct carcinoma. A histologic analysis of 62 resected cases"Ann Surg. 227. 405-411 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Moriguchi T,et al.: "Purification and identification of a major activator for p38 from osmotically shocked cells.Aclivation of MAPKK6 by osmotic shock,tumor necrosis factor-a and H202" J Biol Chem. 271. 26981-26988 (1996)

    • Related Report
      1998 Annual Research Report
  • [Publications] Watanabe Y,et al.: "Characterization of a serum response factor-like protein in saccharomyces cerevisiae,Rlml,which has transcriptional activity regulated by the Mpkl(Slt2)mitogen-activated protein kinase pathway" Mol Cell Biol. 17. 2615-1623 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kamimoto T,et al.: "Dymple,a novel dynamin-like high molecular weight GTPase lacking a proline-rich C-terminal domani in mammalian cells" J Biol Chem. 273. 1044-1051 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shimomura A,et al.: "Dominant negative ATFl blocks cyclic AMP-induced neurite outgrowth in PC12D cells" J Neurochem. 70. 1029-1034 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Nimura Y,et al.: "Aggressive surgical treatment of hilar cholangiocarcinoma" J Hep Bil Pancr Surg. 5. 52-61 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sakamoto E,et al.: "The pattern of infiltration at the proximal A histologic analysis of 62 resected cases." Ann Surg. 227. 405-411 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Watsuji,T., Okamoto,Y., et al: "Controlled gene expression will a reverse tetracycline regulated retroviral vector(RTRV)system." Biochem, Biophys. Res. Commun. 234(3). 769-773 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kamimoto,T., Nagai,Y., et al: "Dymple, a Novel Dynamin-like High Molecular Weight GTP ase Lacking a Proline-rich Carboxyl-terminal Domain in Mammalian Cells." J. Biol. Chem. 273(2). 1044-1051 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nagai,Y., Kojima,T., et al: "Identification of a novel nuclear speokle-type protein, SPOP," FEBS Letters. 418. 23-26 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Muro,Y., Ogawa,Y., et al: "Autoantibody to thioredoxin reductase in an ovarian cancer patient." Biochem. Biophys. Res. Commun.(in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Watanabe,Y., Takaetsu,G., et al: "Characterization of a Serum Response Factor-Like Protein in Saccharomyces cerevisiae, Rlm1, Which Has Transcriptional Activity Regulated by the Mpk1(Slt2)Mitogen-Activated Protein Kinase Pathway." Mol. Cell. Biol.17(5). 2615-2623 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kobayashi,M Shimomura,A., et al: "Phosphorylation of ATF-1 enhances its DNA binding and transcription of the Na, K-ATP ase al subunit gene promotor." Research Nucleic Acid. 25(4). 877-882 (1997)

    • Related Report
      1997 Annual Research Report

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

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