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Administration-route and gender-independent long-term therapeutic correction of phenylketonuria (PKU) in a mouse model by recombinant adeno-associated virus 8 pseudotyped vector-mediated gene transfer

Gene Therapy volume 13, pages 587–593 (2006)Cite this article

Abstract

Phenylketonuria (PKU) is an inborn error of metabolism caused by deficiency of the hepatic enzyme phenylalanine hydroxylase (PAH) which leads to high blood phenylalanine (Phe) levels and consequent damage of the developing brain with severe mental retardation if left untreated in early infancy. The current dietary Phe restriction treatment has certain clinical limitations. To explore a long-term nondietary restriction treatment, a somatic gene transfer approach in a PKU mouse model (C57Bl/6-Pahenu2) was employed to examine its preclinical feasibility. A recombinant adeno-associated virus (rAAV) vector containing the murine Pah-cDNA was generated, pseudotyped with capsids from AAV serotype 8, and delivered into the liver of PKU mice via single intraportal or tail vein injections. The blood Phe concentrations decreased to normal levels (⩽100 μ M or 1.7 mg/dl) 2 weeks after vector application, independent of the sex of the PKU animals and the route of application. In particular, the therapeutic long-term correction in females was also dramatic, which had previously been shown to be difficult to achieve. Therapeutic ranges of Phe were accompanied by the phenotypic reversion from brown to black hair. In treated mice, PAH enzyme activity in whole liver extracts reversed to normal and neither hepatic toxicity nor immunogenicity was observed. In contrast, a lentiviral vector expressing the murine Pah-cDNA, delivered via intraportal vein injection into PKU mice, did not result in therapeutic levels of blood Phe. This study demonstrates the complete correction of hyperphenylalaninemia in both males and females with a rAAV serotype 8 vector. More importantly, the feasibility of a single intravenous injection may pave the way to develop a clinical gene therapy procedure for PKU patients.

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Acknowledgements

We thank N Blau for helpful discussions, D McDonald for the BTBR-Pahenu2 mouse strain, CO Harding for the mouse Pah-cDNA, H Büeler for AAV vector plasmids, G Fischer for tail vein injections, the Vector Core of the Department of Medicine form the University of Pennsylvania for the AAV2/8 plasmid, the mass spectrometry unit for Phe determinations from dried blood spots (H Troxler), CW Heizmann and FH Sennhauser for their continuous support, and A Marti(c)nez for recombinant PAH and anti-PAH antibodies. This work was supported by grants from the Anna Mueller Grocholski and the Swiss National Science Foundations (to BT).

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  1. Division of Clinical Chemistry and Biochemistry, University Children's Hospital Zürich, Zürich, Switzerland

    Z Ding & B Thöny

  2. Department of Visceral and Transplantation Surgery, University Hospital of Zürich, Zürich, Switzerland

    P Georgiev

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  1. Z Ding
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Correspondence to B Thöny.

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We determined blood Phe concentrations with the remaining animals in the experiments shown in Figure 2 after 54 weeks of treatment following intraportal vein application and found for the three females Phe values of 180, 198 and 301 μ M, and after 47 weeks of treatment following tail vain application for the two females Phe values of 301 and 816 μ M.

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Ding, Z., Georgiev, P. & Thöny, B. Administration-route and gender-independent long-term therapeutic correction of phenylketonuria (PKU) in a mouse model by recombinant adeno-associated virus 8 pseudotyped vector-mediated gene transfer. Gene Ther 13, 587–593 (2006). https://doi.org/10.1038/sj.gt.3302684

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