| Project/Area Number |
18591651
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | Research Institute, Osaka Medical Center for Cancer and Cardiovascular Disaeses |
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Principal Investigator |
YAMAMURA Hisako Research Institute, Osaka Medical Center for Cancer and Cardiovascular Disaeses, Department of Molecular Medicine, Assocoate Director (50342994)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Katsuhitto Osaka Medical Center for Cancer and Cardiovascular Diseases, Department of Molecular Medicine, Director (40211338)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥300,000)
Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2006: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | Sarcoma / Malignant mesothelioma / Herpes virus / Gene therapy / Hypoxia |
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| Research Abstract |
We added 57 amino acids polypeptide within the ODD sequence of HIFlalpha, including proline residue at 564, to the amino terminal of HSV-1 ICP4 ptotein. Further, we added nuclear localization signal (NLS) comprising 23 amino acids to the amino terminus of ODD-ICP4.
The CMV-NLS-ODD-ICP4-IRES-LacZ-polyA fragment was inserted into the Stu I site of pKpX2 (J. Virol.62,196-205,1988) (kindly provided by Dr. Weller in Coneticut University), and was linealized by digestion with XhoI. The resulting 11.3kb targeted recombination vector, UL39-CMV-NLS-ODD-ICP4-IRES-LacZ-polyA-UL39 was co-transfected with a mutant HSV-1 DNA, d120 into Vero cells (2.5x105cells/well). We isolated a single plaque of the recombinant HSV-1.
The recombinant HSV-1 preferentially replicated in the hypoxic condition (O2 1%) and showed more potent anti-tumor activity. Expression of ICP4 was also enhanced in the hypoxic condition. The recombinant HSV-1 showed prominent anti-tumor activity against human malignant mesothelioma, leiomyosarcoma and breast cancer by direct injection into the tumors.
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