2007 Fiscal Year Final Research Report Summary
Development of targeted gene therapy to incurable sarcoma and malignant mesothelioma
| Project/Area Number |
18591651
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Orthopaedic surgery
|
|---|
| Research Institution | Research Institute, Osaka Medical Center for Cancer and Cardiovascular Disaeses |
|---|
Principal Investigator |
YAMAMURA Hisako Research Institute, Osaka Medical Center for Cancer and Cardiovascular Disaeses, Department of Molecular Medicine, Assocoate Director (50342994)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Katsuhitto Osaka Medical Center for Cancer and Cardiovascular Diseases, Department of Molecular Medicine, Director (40211338)
|
|---|
| Project Period (FY) |
2006 – 2007
|
| Keywords | Sarcoma / Malignant mesothelioma / Herpes virus / Gene therapy / Hypoxia |
|---|
| 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.
|
