1994 Fiscal Year Final Research Report Summary
Gene therapy for inherited neurodegenerative disease using fetal bone marrows stem cell transplantation.
| Project/Area Number |
05670701
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pediatrics
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| Research Institution | The Jikei Univ.School of Medicine |
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Principal Investigator |
TOKORO Toshiharu Jikei Univ.Dep.of Pediatr.ass.prof., 医学部, 講師 (40112841)
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| Co-Investigator(Kenkyū-buntansha) |
IDA H Jikei Univ.Dep.of Pediatr.investigator senior investigator, 医学部, 助手 (90167255)
OHASHI T Jikei Univ.Dep.of Pediatr., senior, 医学部, 助手 (60160595)
ETO Y Jikei Univ.Dep.of Pediatr., prof., 医学部, 教授 (50056909)
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| Project Period (FY) |
1993 – 1994
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| Keywords | Niemann-Pick disease type C / fetus gene therapy / deficient cholesterol esterification / retroviral vector |
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| Research Abstract |
In this experiment, we tried to elucidate the mechanism of pathogenesis of humann Niemann-Pick disease type C and to have a basic approach for cloning of the defective gene in NPC and to create gene therapy for rat fetus by using of retrovirus vector.
1) We detected the attenuated elevation of cytoplasmic calcium concentration following the uptaking of Low Density Lipoprotein in type C Niemann-Pick fibroblast. Moreover, we found that calcium channel agonist (YC-170) improve the attemuated elevation of calcium concentration and the deficient cholesterol esterification by 40% and 90% of normal control, respectively. These data indicate that the attemuated elevation of cytoplasmic calcim concentration was strongly related to the etiology of humann Niemann-Pick disease type C.
2) We confirmed the synthesis of cholesterol within cells was strongly accelerated in transformed NPC fibroblast by origin defective simian virus 40 and we showed the hypersensitivity of NPC cell lines transformed by SV40 to HMG-CoA reductase inhibitor (Lovastatin, Sinvastatin). Therefor, these cell lines and HMG-CoA reductase inhibitor could be useful for the cloning of the defective gene in NPC.
3) We have the succesful transduction and expression of ASA gene in human MLD fibroblast using a newly developed retroviral vector. This is a first step in evaluating the potential of gene therapy for congenital metabolic disorder. And we tried to have the gene theraly for rat fetus by using this developed retroviral vector transduced with ASA gene. After injection of this vector into amonion vein, we tried to find the expression of ASA gene in several organs such as liver, spleen, and kidny but we could not find any protein expression in these organs by immunohistochemical staining of human ASA antibody. Now we are trying to have gene therapy of MPS mouse fetus by using adeno associated vector which was transduced by alpha-glucuronidase gene.
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