1998 Fiscal Year Final Research Report Summary
Gene therapy for Sly disease
| Project/Area Number |
09670837
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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 T. Jikei Univ., Dept.of Pediatrics assi prof., 医学部・小児科, 講師 (60160595)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI H. Jikei Univ., Dept.of Pediatrics senior investigator, 医学部・小児科, 助手 (90266619)
IDA H. Jikei Univ., Dept.of Pediatrics assi prof., 医学部・小児科, 講師 (90167255)
ETO Y. Jikei Univ., Dept.of Pediatrics prof., 医学部・小児科, 教授 (50056909)
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| Project Period (FY) |
1997 – 1998
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| Keywords | Sly disease / Macrophage / retrovirus / gene therapy / beta-glucuronidase |
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| Research Abstract |
The deficiency of human beta-glucuronidase(HBG) results in MPS type VII(Sly syndrome). In this study, we tested the ability to target macrophages with gene therapy for murine MPS VII.We harvested bone marrow cells from syngeneic normal mice and cultivated in CSF-1 containing medium for 2 weeks. More than 95 % of the cells were double positive for CD18/CD11b and macrophage specific antibody, F4/80 by flowcytometry. We gave 2x10^6 cells to the non-myeloablated Sly mouse. One week post-transplantation, donor cells populated liver and spleen. The HBG activity increased from 0.9*0.7 to 28.4*12.5 u/mg and 0.7*0.4 to 29.7*23.1 u/mg in liver and spleen respectively. However, the pathology was unchanged. The HBG activity in liver and spleen decreased by 5 weeks to 3.7*1.5 and 2.3*0.5 respectively, but the pathological improvements were significant and glycosaminoglycan storage was largely cleared. We assayed tissue glycosaminoglycan content by HPLC method. The contents of dermatan sulfate, hepa
… More
ran sulfate, chondroitin sulfate and hyaluronan decreased almost normal level. Next, the macrophages were collected from Sly mouse by the method above, transduced by HBG expressing retrovirus(MFG-HBG), and given to non-myeloablated Sly mouse. By 5 weeks post-transplantation, HBG activity was only marginally above the control level. However, many HBG positive cells were observed and pathological improvement was significant. The glycosaminoglycan content in liver and spleen was decreased to nearly normal level. We also tried second administration of these genetically modified cells to the Sly mouse. The enzymatic activity in live and spleen increased as same level in initial administration. Finally, we tested the ability of MFG-HBG to transduced human macrophage. We cultivated human macrophages from cord blood using GM-CSF and IL-3 and transduecd by MFG-HBG.Three weeks after, HBG activity increased from 833*56 to 6533*877u/mg. The transgene was in the cells. These data suggest that macrophage transplantation is promising for treatment of murine MPS VII without mycloablation, and macrophage may be a good targets for gene therapy for Sly syndrome. Less
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Research Products
(16 results)
