| Project/Area Number |
18590857
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Respiratory organ internal medicine
|
|---|
| Research Institution | Kyushu University |
|---|
Principal Investigator |
TAKAYAMA Koichi Kyushu University, University hospital, Assistant Professor (50274444)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
HARADA Taishi Kyushu University, Graduate Schood of Medical Sciences, Assistant Professor (10380619)
|
|---|
| Project Period (FY) |
2006 – 2007
|
| Project Status |
Completed (Fiscal Year 2007)
|
| Budget Amount *help |
¥3,760,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥360,000)
Fiscal Year 2007: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2006: ¥2,200,000 (Direct Cost: ¥2,200,000)
|
|---|
| Keywords | Gene Therapy / Adenovirus / Tet on system / Lung Cancer / Cell Vehicle / ウィルスベクター / 制限増殖型アデノウィルス / 薬剤誘導プロモーター |
|---|
| Research Abstract |
The aim of this project is to establish the cell vehicle system for the viral vector carrier. To do this, drug inducible promoter based conditionally replicative adenovirus was made at first. Tetracycline inducible promoter was chosen to switch the promoter activation easily. The tetracycline inducible promoter based replicative adenovirus, AdTRE-E1, was constructed by the replacement of wild type El promoter with tetracycline inducible promoter. AdTRE-E1 proliferated in the infected human fibroblast culture cells (WI38) in the presence of tetracycline in the culture medium. Contrary, the virus was silent in the absence of tetracycline. Therefore, addition of tetracycline in the medium functioned as a switch for viral replication initiation. In the next step, telomerase promoter based replicative adenovirus (AdTERT-E1) was used with the tetracycline inducible system for cancer treatment. WI38 cells were co-infected with AdTRE-E1 and AdTERT-E1 simultaneously, and then tetracycline was added into the culture medium. After 72 hours from infection, the viruses proliferated rapidly and destroyed the infected cells and were released into the medium. Apart of the medium containing viruses was transferred to the intact cancer cell culture flask. AdTERT-E1 infected into the cancer cells and proliferated in the cells independently without tetracycline. As a result, infected WI38 cells carried the viruses in the cells keeping the virus ability of its infectivity. The tetracycline inducible system functioned in the peritoneal space in the nude mouse. When the infected WI38 cells were implanted into the mouse intraperitoneally, viruses proliferated in the peritoneal space after feeding of tetracycline in the drinking water. Therefore, this system functioned in vivo as well as in vitro, and was possible for the cell vehicle system for virus carrier.
|
