Faculty Collaboration Database

Sequencing and characterization of the porcine α-galactosidase A gene: towards the generation of a porcine model for Fabry disease. Mol Biol Rep 2011 Jun;38(5):3145-52

Date

02/05/2010

Pubmed ID

20131008

DOI

10.1007/s11033-010-9985-5

Scopus ID

2-s2.0-80052457781 (requires institutional sign-in at Scopus site)   5 Citations

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.

Author List

Yoshimitsu M, Higuchi K, Fan X, Takao S, Medin JA, Tei C, Takenaka T

Author

Jeffrey A. Medin PhD Professor in the Pediatrics department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Cloning, Molecular
Disease Models, Animal
Fabry Disease
Fibroblasts
Gene Transfer Techniques
Genetic Therapy
Genetic Vectors
HeLa Cells
Humans
Lentivirus
Sus scrofa
Trihexosylceramides
alpha-Galactosidase