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Long-term correction of hyperphenylalaninemia by AAV-mediated gene transfer leads to behavioral recovery in phenylketonuria mice

Gene Therapy volume 11, pages 1081–1086 (2004)Cite this article

Abstract

Classical phenylketonuria (PKU) is a metabolic disorder caused by a deficiency of the hepatic enzyme phenylalanine hydroxylase (PAH). If untreated, accumulation of phenylalanine will damage the developing brain of affected individuals, leading to severe mental retardation. Here, we show that a liver-directed PAH gene transfer brought about long-term correction of hyperphenylalaninemia and behavioral improvement in a mouse model of PKU. A recombinant adeno-associated virus (AAV) vector carrying the murine PAH cDNA was constructed and administered to PAH-deficient mice (strain PAHenu2) via the portal vein. Within 2 weeks of treatment, the hyperphenylalaninemic phenotype improved and completely normalized in the animals treated with higher vector doses. The therapeutic effect persisted for 40 weeks in male mice, while serum phenylalanine concentrations in female animals gradually returned to pretreatment levels. Notably, this long-term correction of hyperphenylalaninemia was associated with a reversal of hypoactivity observed in PAHenu2 mice. While locomotory activity over 24 h and exploratory behavior were significantly decreased in untreated PAHenu2 mice compared with the age-matched controls, these indices were completely normalized in 12-month-old male PKU mice with lowered serum phenylalanine. These results demonstrate that AAV-mediated liver transduction ameliorated the PKU phenotype, including central nervous system dysfunctions.

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Acknowledgements

We are grateful to Dr JA Chiorini for pAAV5LacZ and 5RepCapA, Dr J Miyazaki for pCAGGS, Dr T Shiga for PAHenu2 mice, Avigen for pLadeno1, and Advantec Toyo for #545 filter paper. We also thank Dr Y Hakamata for technical assistance in the animal experiments. This work was supported in part by grants from the Ministry of Education, Culture, Sports, Science and Technology, and the Ministry of Health, Labor and Welfare, Japan.

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Authors and Affiliations

  1. Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical School, Tochigi, Japan

    S Mochizuki, H Mizukami, T Ogura, T Okada, K Ozawa & A Kume

  2. Department of Pediatrics, Tokyo Medical University, Tokyo, Japan

    S Mochizuki & A Hoshika

  3. Department of Medical Genetics, Tohoku University Graduate School of Medicine, Sendai, Japan

    S Kure, A Ichinohe, K Kojima & Y Matsubara

  4. Division of Organ Replacement, Center for Molecular Medicine, Jichi Medical School, Tochigi, Japan

    E Kobayahi

  5. Division of Hematology, Department of Medicine, Jichi Medical School, Tochigi, Japan

    K Ozawa

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  1. S Mochizuki
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Mochizuki, S., Mizukami, H., Ogura, T. et al. Long-term correction of hyperphenylalaninemia by AAV-mediated gene transfer leads to behavioral recovery in phenylketonuria mice. Gene Ther 11, 1081–1086 (2004). https://doi.org/10.1038/sj.gt.3302262

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  • DOI: https://doi.org/10.1038/sj.gt.3302262

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