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Lysosomal delivery of therapeutic enzymes in cell models of Fabry disease

  • Original Article
  • Published:
Journal of Inherited Metabolic Disease

Abstract

The success of enzymatic replacement in Gaucher disease has stimulated development of targeted protein replacement for other lysosomal disorders, including Anderson-Fabry disease, which causes fatal cardiac, cerebrovascular and renal injury: deficiency of lysosomal α-Galactosidase A induces accumulation of glycosphingolipids. Endothelial cell storage was the primary endpoint in a clinical trial that led to market authorization. Two α-Galactosidase A preparations are licensed worldwide, but fatal outcomes persist, with storage remaining in many tissues. We compare mechanisms of uptake of α -Galactosidase A into cells relevant to Fabry disease, in order to investigate if the enzyme is targeted to the lysosomes in a mannose-6-phosphate receptor dependent fashion, as generally believed. α -Galactosidase A uptake was examined in fibroblasts, four different endothelial cell models, and hepatic cells in vitro. Uptake of europium-labeled human α -Galactosidase A was measured by time-resolved fluorescence. Ligand-specific uptake was quantified in inhibitor studies. Targeting to the lysosome was determined by precipitation and by confocal microscopy. The quantity and location of cation-independent mannose-6-phosphate receptors in the different cell models were investigated using confocal microscopy. Uptake and delivery of α -Galactosidase A to lysosomes in fibroblasts is mediated by the canonical mannose-6-phosphate receptor pathway, but in endothelial cells in vitro this mechanism does not operate. Moreover, this observation is supported by a striking paucity of expression of cation independent mannose-6-phosphate receptors on the plasma membrane of the four endothelial cell models and by little delivery of enzyme to lysosomes, when compared with fibroblasts. If these observations are confirmed in vivo, alternative mechanisms will be needed to explain the ready clearance of storage from endothelial cells in patients undergoing enzyme replacement therapy.

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Acknowledgments

We thank the Society for Mucopolysaccharide Diseases for essential moral and financial support for the project. This work was partly supported by grants from the Cambridge Biomedical Research Centre (Metabolic Theme) funded by the National Institute of Health Research, UK. We wish to thank Prof. R. Schiffmann and Dr. C. R. Kaneski who kindly provided the Immortalized Fabry Endothelial cell line-1 (IMFE-1), Maria del Mar Roldan Ortiz for her expert technical assistance in cell culturie and Prof. N Morrell and Dr. Xudong Yang for providing us with Human Pulmonary Arthery Endothelial cells (HPAEC).

Conflict of interest

This work was partly supported by the Society for Mucopolysaccharide Diseases.

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  1. Department of Medicine Addenbrooke’s Hospital, University of Cambridge, Hills Road, Cambridge, CB2 0QQ, UK

    D. Marchesan, T. M. Cox & P. B. Deegan

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  1. D. Marchesan
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  2. T. M. Cox
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  3. P. B. Deegan
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Correspondence to P. B. Deegan.

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Communicated by: Robin Lachmann

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Marchesan, D., Cox, T.M. & Deegan, P.B. Lysosomal delivery of therapeutic enzymes in cell models of Fabry disease. J Inherit Metab Dis 35, 1107–1117 (2012). https://doi.org/10.1007/s10545-012-9472-3

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  • DOI: https://doi.org/10.1007/s10545-012-9472-3

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