1995 Fiscal Year Final Research Report Summary
Gene therapy of urea cycle deficirncy
| Project/Area Number |
06454610
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Human genetics
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| Research Institution | Kumamoto University |
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Principal Investigator |
MATSUDA Ichiro Kumamoto University School of Medicine Department of Pediatrics Professor, 医学部, 教授 (10000986)
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| Co-Investigator(Kenkyū-buntansha) |
SAITO Izumu The University of Tokyo The Institute of Medical Science Laboratory of Molecular, 医科学研究所, 助教授 (70158913)
INDO Yasuhiro Kumamoto University School of Medicine Department of Hospital Assistant Professo, 医学部・附属病院, 助手 (40244131)
ENDO Fumio Kumamoto University School of Medicine Department of Hospital Lecturer, 医学部・附属病院, 講師 (00176801)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Gene therapy / Urea cycle / Ornithine transcarbamylase / Adenovirus vector |
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| Research Abstract |
Ornithine transcarbamylase (OTC) deficiency, the most commom and severe inborn error of the urea cycle in humans, remains without adequate treatment, and ortality rates are high. Adenoviral vectors provide an efficient system for gene delivery, but there are problems, including toxicity. Efficient promoters that reduce the amount of vector required for treatment need to be developed. We constructed two recombinant adenoviral vectors, AdexCAGhOTC and AdexSRalphahOTC,which harbor the human OTC gene under transcriptional control of CAG (a modified chicken beta-actin promoter with CMV-IE enhancer) and SRalpha (the SV 40 early prommoter with the R segment and part of the U5 segment of the HTLV-1 LTR) , respectively. Each was tested in adult spf^<ash> mice, an animal model of human OTC deficiency, and in primary human hepatocytes with OTC deficiency. Spf^<ash> mice have a pronounced orotic aciduria as seen in humans. A complete recovery of hepatic OTC activity with minimal tissue damage was observed in these animals following the intravenous administration of AdexCAGhOTC alone. Western blot analysis confirmed hepatic OTC expression and normalization of orotic aciduria was evident for 60 days. Enzyme activities of primary human hepatocytes infected with AdexCAGhOTC were 10-40 times higher than those with AdexSRalphahOTC.Thus, the adenoviral vector with an efficient promoter such as CAG,can be given further considerartion for possible gene therapy in humans with OTC deficiency.
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