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Molecular basis for globotriaosylceramide regulation and enzyme uptake in immortalized aortic endothelial cells from Fabry mice

  • Original Article
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Journal of Inherited Metabolic Disease

Abstract

Fabry disease is caused by deficient activity of α-galactosidase A and subsequent intracellular accumulation of glycosphingolipids, mainly globotriaosylceramide (Gb3). Vascular endothelial cells may play important roles in disease pathogenesis, and are one of the main target cell types in therapeutic interventions. In this study, we generated immortalized aortic endothelial cell lines from a mouse model of Fabry disease. These cells retained endothelial cell-specific markers and functions. Gb3 expression level in one of these clones (referred to as FMEC2) was highly susceptible to culture media, and appeared to be regulated by glucosylceramide synthase. Results also showed that Gb3 could be upregulated by hydrocortisone. FMEC2 express the mannose 6-phosphate receptor and sortilin but not the mannose receptor. Uptake studies suggested that sortilin plays a role in the binding and internalization of mammalian cell-produced α-galactosidase A. Moss-aGal (a plant-made enzyme) was endocytosed by FMEC2 via a receptor other than the aforementioned receptors. In conclusion, this study suggests that glucosylceramide synthase and hydrocortisone may play important roles in modulating Gb3 levels in Fabry mouse aortic endothelial cells, and that endocytosis of recombinant α-galactosidase A involves a combination of multiple receptors depending on the properties of the enzyme.

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Acknowledgments

We thank Dilechia Hawthorne (Institute of Metabolic Disease, Dallas) for her technical assistance during the study. This work was supported by Baylor Research Institute.

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Authors and Affiliations

  1. Institute of Metabolic Disease, Baylor Research Institute, 3812 Elm Street, Dallas, TX, 75226, USA

    Xing-Li Meng, Taniqua S. Day, Nathan McNeill, Paula Ashcraft, Jin-Song Shen & Raphael Schiffmann

  2. Greenovation Biotech GmbH, Freiburg, Germany

    Thomas Frischmuth

  3. Sanofi Genzyme, 49 New York Avenue, Framingham, MA, 01701, USA

    Seng H. Cheng

  4. Department of Internal Medicine, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX, 75390, USA

    Zhi-Ping Liu

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  1. Xing-Li Meng
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  2. Taniqua S. Day
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  3. Nathan McNeill
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  4. Paula Ashcraft
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  7. Zhi-Ping Liu
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  8. Jin-Song Shen
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  9. Raphael Schiffmann
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Correspondence to Jin-Song Shen.

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Conflict of interest

Xing-Li Meng, Taniqua S. Day, Nathan McNeill, Paula Ashcraft, Zhi-Ping Liu, and Jin-Song Shen declare that they have no conflict of interest. Thomas Frischmuth is employee in Greenovation Biotech. Seng H. Cheng is employee in Genzyme. Raphael Schiffmann received research funds and honoraria in the past 5 years from Genzyme, Shire and Amicus Therapeutics.

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Communicated by: Robin Lachmann

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Meng, XL., Day, T.S., McNeill, N. et al. Molecular basis for globotriaosylceramide regulation and enzyme uptake in immortalized aortic endothelial cells from Fabry mice. J Inherit Metab Dis 39, 447–455 (2016). https://doi.org/10.1007/s10545-016-9920-6

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  • DOI: https://doi.org/10.1007/s10545-016-9920-6

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