| Project/Area Number |
07670905
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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 |
Pediatrics
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| Research Institution | The Jikei University School of Medicine |
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Principal Investigator |
OHASHI Touya The Jikei University School of Medicine assi.prof., 小児科, 講師 (60160595)
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| Co-Investigator(Kenkyū-buntansha) |
SUGAMA Seiichi The Jikei University School of Medicine senior investigator, 小児科, 助手 (10154452)
ETO Yoshikatu The Jikei University School of Medicine prof., 小児科, 助手 (50056909)
IDA Hiroyuki The Jikei University School of Medicine assi.prof., 小児科, 講師 (90167255)
津田 隆 東京慈恵会医科大学, 医学部・小児科学, 助手 (50188554)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1995: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | Sly disease / retrovirus / beta-glucuronidase / adenovirus / CD34+ cells / transduction / NOD-SCIDマウス / LTCIC(Long Term Culture Iniciating Cell) / ムコ多糖症VII型 / ヒト骨髄細胞 / β-glucuronidase遺伝子 / CD34+細胞 / サザンブロット法 / 骨髄前駆細胞 / モデルマウス / ムコ多糖症 / 遺伝子治療 |
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| Research Abstract |
Mucopolysaccharidosis type VII (Sly disease) is a lysosomal storage disease caused by inherited deficiency of the lysosomal enzyme beta-glucuronidase. A murine model of this disorder has been well characterized and used to study a number of forms of experimental therapies including gene therapy. In this study, we generated recombinant adenovirus which express human beta-glucuronidase and administered this recombinant adenovirus to model mice intravenously. The beta-glucuronidase activities in liver and spleen were elevated and expression persisted for at least 35 days. Pathological adnormalities of these tissues were improved. Following administration of the recombinant adenovirus directly into the lateral ventricles of mutant mice, the recombinant adenovirus infected mainly ependymal cells and choroid. Only a small number of brain parenchymal cells were infected.
As an alternative approach, we also study transfer of the human beta-glucuronidase gene to human hematopoietic cells. Bone marrow transplantation in mucopolysaccharidosis patients in early clinical stages has been reported to halt the progression of neurological deterioration. As a first step, we generated recombinant retrovirus which express human beta-glucuronidase gene and infected this virus to CD34+ cells purified from umbilical cord blood. Using a centrifugation promoted infection protocol, we got transduction efficiency (Ave.67%) in colony forming unit granulocyte/macrophage and burst forming unit erythrocyte. Experiments are underway whether transduced CD34+ cells migrate to brain in severe combined immuno-deficient mice.
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