Abstract
Phenylketonuria (PKU) is caused by deficiency of phenylalanine hydroxylase (PAH) in the liver. Patients with PKU show increased L-phenylalanine in blood, which leads to mental retardation and hypopigmentation of skin and hair. As a step toward gene therapy for PKU, we constructed a replication-defective, E1/E3-deleted recombinant adenovirus harboring human PAH cDNA under the control of a potent CAG promoter. When a solution containing 1.2 × 109 plaque-forming units of the recombinant adenovirus was infused into tail veins of PKU model mice (Pahenu2), predominant expression of PAH activity was observed in the liver. The gene transfer normalized the serum phenylalanine level within 24 h. However, it also provoked a profound host immune response against the recombinant virus; as a consequence, the biochemical changes lasted for only 10 d and rechallenge with the virus failed to reduce the serum phenylalanine concentration. Administration of an immunosuppresant, FK506, to mice successfully blocked the host immune response, prolonged the duration of gene expression to more than 35 d, and allowed repeated gene delivery. We noted a change in coat pigmentation from grayish to black after gene delivery. The current study is the first to demonstrate the reversal of hypopigmentation, one of the major clinical phenotypes of PKU in mice as well as in humans, by adenovirus-mediated gene transfer, suggesting the feasibility of gene therapy for PKU.
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Abbreviations
- PKU:
-
phenylketonuria
- PAH:
-
phenylalanine hydroxylase
- DMEM:
-
Dulbecco modified Eagle medium
- FBS:
-
fetal bovine serum
- pfu:
-
plaque-forming units
- ALT:
-
alanine aminotransferase
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Acknowledgements
The authors thank Dr. Alexandra Shedlovsky for a gift of Pahenu2 and critical reading of the manuscript. They also thank Dr. Jun-ichi Miyazaki for providing the CAG promoter and pUC-CAGGS and Ms. Kumiko Osawa for technical assistance.
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This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports and Science of Japan and grants from the Ministry of Health and Public Welfare of Japan.
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Nagasaki, Y., Matsubara, Y., Takano, H. et al. Reversal of Hypopigmentation in Phenylketonuria Mice by Adenovirus-Mediated Gene Transfer. Pediatr Res 45, 465–473 (1999). https://doi.org/10.1203/00006450-199904010-00003
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DOI: https://doi.org/10.1203/00006450-199904010-00003
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