| Project/Area Number |
09470145
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Respiratory organ internal medicine
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| Research Institution | Tohoku University |
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Principal Investigator |
YAEKASHIWA Masahiro Institute of Development, Aging and Cancer, Tohoku University, Assistant Professor, 加齢医学研究所, 助手 (70261477)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Toshikazu Division of Biochemistry, Biomedical Research Center, Osaka University Medical S, 医学部, 教授 (00049397)
SATOH Ken Institute of Development, Aging and Cancer, Tohoku University, Associate Profess, 加齢医学研究所, 助教授 (00215782)
NUKIWA Toshihiro Institute of Development, Aging and Cancer, Tohoku University, Professor, 加齢医学研究所, 教授 (40129036)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥12,600,000 (Direct Cost: ¥12,600,000)
Fiscal Year 1998: ¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 1997: ¥8,900,000 (Direct Cost: ¥8,900,000)
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| Keywords | HGF / Gene therapy / Bleomycin / Transgene / Lung Fibrosis |
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| Research Abstract |
Hepatocyte growth factor (HGF) is a pleiotropic factor that plays an essential role in lung repair and regeneration after injury. In this project, we tried to obtain adequate expression of HGF from transgene in vivo for therapy of lung fibrosis that follows lung injury.
Methods
1. The expression of HGF protein was examined when human HGF cDNA was introduced into cultured cells with pCI-neo Mammalian Expression vector.
2. The expression of HGF protein was examined when rat HGF cDNA was introduced into mice with adenoviral vector (Adex1CAHHGF : constructed with rat HGF gene under control of GAG promoter). Therapeutic effects of HGF protein from transgene was evaluated in mouse models of lung fibrosis induced by bleomycin (BLM).
3. The mechanism of therapeutic effects of HGF on lung fibrosis was analyzed in vitro and in vivo.
Results
1. Human HGF cDNA was introduced in culture cell lines (A549 cell, 293 cell) with pCI-neo mammalian expression vector. The expression of HGF protein was confirmed in the supernatant and transduced cells by ELISA.The transduced cells expressed HGF constantly.
2. High expression of HGF protein and mRNA in liver was observed in mice after intraperitoneal administration of Adex1CAHHGF.The concentration of HGF in liver of mice treated with adenoviralvector elevated at day 3 (286.0 ng/g tissue) and returned to normal level at day 7 (119.9 ng/g tissue). The concentration of HGF in lung moved as that in liver. The lung fibrosis by BLM was suppressed with administration of Adex1CAHHGF.Treatment with HGF resulted in resistance to Staurosporin- and BLM-induced apoptotic cell death in vitro.
Administration of HGF showed anti-TGF-beta1 effects in vivo.
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