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Early gestational gene transfer with targeted ATP7B expression in the liver improves phenotype in a murine model of Wilson's disease

Abstract

The ideal gene therapy for metabolical liver disorders would target hepatocytes before the onset of disease and be durable, non-toxic and non-immunogenic. Early gestational gene transfer can achieve such goals. Here, we demonstrate that prenatal gene transfer of human Atp7b reduces liver pathology and improves biochemical markers in Atp7b−/− mice, a murine model of Wilson's disease (WD). Following prenatal injection of lentivirus vector containing the human Atp7b gene under the transcriptional control of a liver-specific promoter, the full-length ATP7B was detectable in mouse livers for the entire duration of experiments (20 weeks after birth). In contrast to a marked pathology in non-injected animals, livers from age-matched treated mice consistently demonstrated normal gross and histological morphology. Hepatic copper content was decreased in the majority of treated mice, although remaining copper levels varied. Improvement of hepatic copper metabolism was further apparent from the presence of copper-bound ceruloplasmin in the sera and normalization of the mRNA levels for HMG CoA-reductase. With this approach, the complete loss of copper transport function can be ameliorated, as evident from phenotypical improvement in treated Atp7b−/− mice. This study provides proof of principle for in utero gene therapy in WD and other liver-based enzyme deficiencies.

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Abbreviations

WD:

Wilson's disease

ATP7B:

adenosine triphosphate 7B

LSP:

liver-specific promoter

GFP:

green fluorescent protein

MND:

modified myeloid proliferative sarcoma virus promoter

HMG CoA reductase:

3-hydroxy-3-methyl-glutaryl-CoA reductase

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Acknowledgements

The authors thank Dr Jeffrey M Bergelson (Children's Hospital of Philadelphia) for the enterocyte line Caco-2, Dr David Baltimore (California Institute of Technology) for the HIV-1-based transfer vector, a modified FG12 plasmid, Dr Katherine A High (Children's Hospital of Philadelphia) for the liver-specific promoter, Archana Bora (Children's Hospital of Philadelphia) for help with qRT-PCR, and Keith Alcorn (Children's Hospital of Philadelphia) for help with animal husbandry, and Dr Ann Hubbard (Johns Hopkins University) for helpful discussions. JLR was supported by grant 5T32HD46402 from the National Institutes of Health (AWF); SL and LG were supported by grant P01 GM067166 from the National Institutes of Health to SL. Additional support was provided by funds from the Ruth and Tristram C Colket, Jr Chair of Pediatric Surgery (AWF).

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Correspondence to A W Flake.

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Roybal, J., Endo, M., Radu, A. et al. Early gestational gene transfer with targeted ATP7B expression in the liver improves phenotype in a murine model of Wilson's disease. Gene Ther 19, 1085–1094 (2012). https://doi.org/10.1038/gt.2011.186

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