| Project/Area Number |
11694309
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Hematology
|
|---|
| Research Institution | Jichi Medical School |
|---|
Principal Investigator |
KUME Akihiro Jichi Medical School, School of Medicine, Associate Professor, 医学部, 助教授 (10264293)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MIZUKAMI Hiroaki Jichi Medical School, School of Medicine, Assistant Professor, 医学部, 助手 (20311938)
HANAZONO Yutaka Jichi Medical School, School of Medicine, Assistant Professor, 医学部, 講師 (70251246)
OZAWA Keiya Jichi Medical School, School of Medicine, Professor, 医学部, 教授 (30137707)
OKADA Takashi Jichi Medical School, School of Medicine, Assistant Professor, 医学部, 助手 (00326828)
間野 博行 自治医科大学, 医学部, 助教授 (90240704)
卜部 匡司 自治医科大学, 医学部, 助手 (40213516)
|
|---|
| Project Period (FY) |
1999 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2000: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1999: ¥2,400,000 (Direct Cost: ¥2,400,000)
|
|---|
| Keywords | gene therapy / hematopoietic stem cells / chronic granulomatous disease / selective amplifier gene / レトロウイルスベクター |
|---|
| Research Abstract |
In order to improve the efficacy of hematopoietic stem cell gene therapy, a retroviral vector with high transduction/expression efficiency and an in vivo expansion system of transduced hematopoietic cells were developed. 1) A new retroviral vector (MGK) was constructed, by assembling the long terminal repeats and the primer tRNA binding site from MSCV and the packaging and splicing signals of gag-killed MFG. 2) For in vivo expansion of transduced hematopoietic cells, a novel system named 'selective amplifier genes' was developed. Murine bone marrow cells were transduced with a bicistronic retroviral vector containing a tamoxifen-responsive selective amplifier gene and the EGFP marker gene, and transplanted to lethally irradiated hosts. After hematopoietic reconstitution, a subset of the recipients were challenged with tamoxifen. The challenged animals showed significantly higher frequency of EGFP-expressing cells than the control animals. The results indicated that the transduced hematopoietic stem/progenitor cells were expanded by the selective amplifier gene/tamoxifen system, and in vivo expansion of the gene-modified cells is feasible.
These results were presented at the annual meetings of American Society of Gene Therapy and the American Society of Hematology. We invited Dr. M. C. Dinauer from Indiana University, U.S.A., to discuss about their clinical gene transfer trial for X-linked chronic granulomatous disease. We also discussed with collaborators in Tokyo, Nagasaki and Kumamoto to set up a clinical protocol of gene therapy for chronic granulomatous disease.
|
