| Project/Area Number |
16590960
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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 |
Hematology
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| Research Institution | JICHI MEDICAL UNIVERSITY |
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Principal Investigator |
TAKATOKU Masaaki JICHI MEDICAL UNIVERSITY, School of Medicine, Assistant Professor, 医学部, 講師 (20285787)
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| Co-Investigator(Kenkyū-buntansha) |
UEDA Masuzu JICHI MEDICAL UNIVERSITY, School of Medicine, Instructor, 医学部, 助手 (60364501)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2005: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | gene therapy / CGD / SAG / iBMT / 造血幹細胞移植 |
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| Research Abstract |
1.Gene transduction and Transplantation experiment to the target cells.
1)The optimization of the transplantation preconditioning.
To examine which is better methods for high engraftment efficiency under no preconditioning using intra tail vein bone marrow transplantation (IV-BMT) or intra bone marrow transplantation (iBMT). Our results demonstrated that it was no significant difference of engraftment efficiency between IV-BMT and iBMT. Both methods were also very low-efficiency.
Therefore, we used non-myeloablative irradiation (2Gy) as preconditioning. Then we compared engraftment efficiency with IV-BMT or iBMT. As this result, the engraftments were confirmed in both methods, and also we detected high expression of the treatment gene. (reactive oxygen production rate : IV-BMT 10%,iBMT 30%)
2)in vivo amplification by the selective amplified gene (SAG).
We developed the vector which included treatment gene (GP91^<phox>) in retrovirus vector mounted SAG. The transduced cells were transplanted in the mouse model, and the gene system action was confirmed.
As this result, the engraftment was confirmed in the mouse which did iBMT after non-myeloablative irradiation, and in addition, the peripheral blood cell population which introduced the treatment gene were increased after the Erythropoietin (EPO) administration as molecular switch of SAG. But this increase was the transience, and it was the EPO stimulus dependence.
2.The safety of this system.
We analyzed gene integration site of the retrovirus using the linear amplified mediated polymerase chain reaction (LAM-PCR) method.
The random integration of the vector virus was confirmed without increase bits of the specific integration site as a result of tracking the blood cell in our mouse models.
It will be observed in the long term in future too.
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