2005 Fiscal Year Final Research Report Summary
Targeted gene therapy for incurable sarcoma using a novel replication-selective and oncolytic herpes virus harboring GM-CSF gene
| Project/Area Number |
16591521
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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 | Osaka Medical Center for Cancer & Cardiovascular Diseases |
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Principal Investigator |
YAMAMURA Hisako Osaka Medical Center for Cancer & Cardiovascular Diseases, Department of Molecular Medicine, Associate director, 研究所, 主任研究員 (50342994)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Katsuhito Osaka Medical Center for Cancer & Cardiovascular Diseases, Department of Molecular Medicine, Director, 研究所, 部長 (40211338)
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| Project Period (FY) |
2004 – 2005
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| Keywords | herpes simplex virus / calponin / oncolytic virus / gene therapy / sarcoma / GM-CSF |
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| Research Abstract |
We have previously described a thymidme kinase (TK)-defective type I herpes simplex virus (HSV-1) mutant d12.CALP in which smooth muscle-specific calponin promoter drives expression of the RS1 gene encoding an essential trans-activating factor (ICP4) for viral genes (Yamamura et al. Cancer Res. 61; 3969-3977,2001). In order to improve efficacy and safety for pre-clinical and clinical testing, we have developed a new conditionally replicating oncolytic HSV-1 (d12.CALPΔRR GM-CSF) in which smooth muscle-specific calponin transcriptional regulatory sequence drives the RS1 gene and the human GM-CSF cDNA via bicistronic expression using the internal ribosomal entry site (IRES2). The engineered HSV-1 is a derivative of ICP4-null mutant d120 carrying the intact TK gene and an insertional mutation in the U_L39 gene encoding a large subunit of ribonucleotide reductase (ICP6), an essential enzyme for viral replication. d12.CALPΔRR GM-CSF also contains the Escherichia coli lacZ gene under the cont
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rol of the intrinsic U_L39 gene promoter to trace viral replication. We isolated three clones of the d12.CALPΔRR GM-CSF using limited dilution methods after infection to the SK-LMS-1 human leiomyosarcoma cells, following the selection of two cycles of infection to ICP4-transfected Vero cells. We also investigated the replication of the d12.CALPΔRR in ICP4-transfected Vero cells in serum free medium VP-SFM (Invitrogen), and successfully carried out a large scale viral purification using this medium. The sensitivity to GCV of the d12.CALPΔRR virus in VP-SFM was comparable to that in 10%FBS/DMEM medium. We further confirmed that replication of the d12.CALPΔRR virus in MethA fibrosarcoma cells in immuno-competent mice could induce MethA-specific anti-tumor immunity. We suggest that in addition to a direct oncolytic effects on the tumors, novel anti-leiomyosarcoma agents d12.CALPΔRR and d12.CALPΔRR GM-CSF may destroy sarcoma tumors in remote organs via systemic anti-tumor immunity, and thus should be investigated further as a challenging therapy for metastasis of incurable human sarcoma tumors Less
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Research Products
(4 results)
