1997 Fiscal Year Final Research Report Summary
GENE THERAPY ON HEPATIC ENZYME DEFICIENCY.
| Project/Area Number |
08457218
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | TOHOKU UNIVERSITY |
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Principal Investigator |
NARISAWA Kuniaki TOHOKU UNIVERSITY,SCHOOL OF MEDICINE,PROFESSOR, 医学部, 教授 (90004647)
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| Co-Investigator(Kenkyū-buntansha) |
KURE Shigeo TOHOKU UNIVERSITY,SCHOOL OF MEDICINE,RESEARCH ASSOCIATE, 医学部, 助手 (10205221)
MATSUBARA Yoichi TOHOKU UNIVERSITY,SCHOOL OF MEDICINE,ASSOCIATE PROFESSOR, 医学部, 助教授 (00209602)
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| Project Period (FY) |
1996 – 1997
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| Keywords | HEPATIC ENZYME DEFICIENCY / PHENYLKETONURIA / GENE THERAPY / ADNOVIRUS VECTOR / SERUM PHENYLALANINE / COAT COLOR |
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| Research Abstract |
We investigated gene therapy on hepatic enzyme deficiency using PKU model mice. We constructed a replication-defective recombinant adenovirus harboring human PAH cDNA under the control of a potent CAG promoter using a cosmid-cassette method. Infection of COS7 cells with the recombinant virus at m.o.i=3.0 in vitro produced the PAH activity equivalent to normal hepatocytes. When the solution containing 1.2x10^9 p.f.u of the virus was infused into the tail vein of PKU model mice, PAH cDNA and enzymatic PAH activity were mainly detected in liver. Serum phenylalanine concentration decreased to normal level within 24 hrs. However, the biochemical change lasted for only 10 days and re-administration of the virus failed to correct hyperphenylalaninemia due to host immune response against the adenovirus. When the mice were treated with daily administration of an immunosuppressant FK506, the duration of gene expression was prolonged to more than 35 days and repeated gene delivery was able to decrease the serum phenylalanine level. In these experiments, hypopigmented PKU mice showed distinct pigmentation of coat, from grayish color to black, regardless of FK506 treatment. The phenotypic reversal was presumably due to improved tyrosine metaboLism. The duration of hypopigmentation closely correlated with that of the normalization of phenylalanine level. Our study is the first to demonstrate the collection of physical phenotype in PKU mice by gene transfer, suggesting the feasibility of gene therapy on PKU.
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