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Sequencing and characterization of the porcine α-galactosidase A gene: towards the generation of a porcine model for Fabry disease

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Abstract

Fabry disease is an inherited lysosomal disorder caused by a deficiency of alpha-galactosidase A (α-gal A). The systemic accumulation of substrate, mainly globotriaosylceramide (Gb3), results in organ failure. Although Gb3 accumulation has been observed in an α-gal A-deficient mouse model, important clinical manifestations were not seen. The pursuit of effective treatment for Fabry disease through gene therapy, for example, has been hampered by the lack of a relevant large animal model to assess the efficacy and safety of novel therapies. Towards assembling the tools to generate an alternative animal model, we have sequenced and characterized the porcine ortholog of the α-gal A gene. When compared to the human α-gal A, the porcine α-gal A showed a high level of homology in the coding regions and located at chromosome Xq22. Cell lysate and supernatants from Fabry patient-derived fibroblasts transduced with a lentiviral vector (LV) carrying the porcine α-gal A cDNA (LV/porcine α-gal A), showed high levels of α-gal A activity and its enzymological stability was similar to that of human α-gal A. Uptake of secreted porcine α-gal A was observed into non-transduced cells and was partially inhibited by soluble mannose-6-phosphate. Furthermore, Gb3 accumulation was reduced in Fabry patient-derived fibroblasts transduced with the LV/porcine α-gal A. In conclusion, we elucidated and characterized the porcine α-gal A gene and enzyme. Similarity in enzymatic profile and chromosomal location between α-gal A of porcine and human origins may be of great advantage for the development of a large animal model for Fabry disease.

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Acknowledgements

We thank Miss Takane Takemoto and Miss Miho Hachiman for their technical help. This research was supported in part by a Grant-in-Aid for Idiopathic Cardiomyopathy Research Committee from the Ministry of Health, Labour and Welfare of Japan.

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Authors and Affiliations

  1. Division of Cardiac Repair and Regeneration, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima, 890-8520, Japan

    Makoto Yoshimitsu, Koji Higuchi, Chuwa Tei & Toshihiro Takenaka

  2. Division of Stem Cell and Developmental Biology, Ontario Cancer Institute, Toronto, ON, M5G-2M1, Canada

    Xin Fan & Jeffrey A. Medin

  3. Frontier Science Research Center, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima, 890-8520, Japan

    Sonshin Takao

  4. Department of Medical Biophysics and the Institute of Medical Science, University of Toronto, Toronto, ON, M5G-2M1, Canada

    Jeffrey A. Medin

  5. Department of Cardiovascular, Respiratory and Metabolic Medicine, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima, 890-8520, Japan

    Chuwa Tei

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  1. Makoto Yoshimitsu
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  2. Koji Higuchi
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  3. Xin Fan
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  4. Sonshin Takao
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  5. Jeffrey A. Medin
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  7. Toshihiro Takenaka
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Correspondence to Toshihiro Takenaka.

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Makoto Yoshimitsu and Koji Higuchi authors contributed equally to this work.

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Yoshimitsu, M., Higuchi, K., Fan, X. et al. Sequencing and characterization of the porcine α-galactosidase A gene: towards the generation of a porcine model for Fabry disease. Mol Biol Rep 38, 3145–3152 (2011). https://doi.org/10.1007/s11033-010-9985-5

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