title: Molecular basis for globotriaosylceramide regulation and enzyme uptake in immortalized aortic endothelial cells from Fabry mice.
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.
authors.affiliation: Institute of Metabolic Disease, Baylor Research Institute, 3812 Elm Street, Dallas, TX, 75226, USA.
authors.forename: Xing-Li
authors.initials: XL
authors.lastname: Meng
authors.affiliation: Institute of Metabolic Disease, Baylor Research Institute, 3812 Elm Street, Dallas, TX, 75226, USA.
authors.forename: Taniqua S
authors.initials: TS
authors.lastname: Day
authors.affiliation: Institute of Metabolic Disease, Baylor Research Institute, 3812 Elm Street, Dallas, TX, 75226, USA.
authors.forename: Nathan
authors.initials: N
authors.lastname: McNeill
authors.affiliation: Institute of Metabolic Disease, Baylor Research Institute, 3812 Elm Street, Dallas, TX, 75226, USA.
authors.forename: Paula
authors.initials: P
authors.lastname: Ashcraft
authors.affiliation: Greenovation Biotech GmbH, Freiburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Frischmuth
authors.affiliation: Sanofi Genzyme, 49 New York Avenue, Framingham, MA, 01701, USA.
authors.forename: Seng H
authors.initials: SH
authors.lastname: Cheng
authors.affiliation: Department of Internal Medicine, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX, 75390, USA.
authors.forename: Zhi-Ping
authors.initials: ZP
authors.lastname: Liu
authors.affiliation: Institute of Metabolic Disease, Baylor Research Institute, 3812 Elm Street, Dallas, TX, 75226, USA. Jinsong.Shen@baylorhealth.edu.
authors.forename: Jin-Song
authors.initials: JS
authors.lastname: Shen
authors.affiliation: Institute of Metabolic Disease, Baylor Research Institute, 3812 Elm Street, Dallas, TX, 75226, USA.
authors.forename: Raphael
authors.initials: R
authors.lastname: Schiffmann
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism; D000818:Animals; D001011:Aorta / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D002460:Cell Line; D004195:Disease Models, Animal; D004705:Endocytosis / Q000502:physiology; D042783:Endothelial Cells / Q000201:enzymology* / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000201:enzymology / Q000378:metabolism*; D000795:Fabry Disease / Q000201:enzymology* / Q000378:metabolism*; D005964:Glucosyltransferases / Q000378:metabolism; D006028:Glycosphingolipids / Q000378:metabolism; D037181:Lectins, C-Type / Q000378:metabolism; D008297:Male; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D017527:Receptor, IGF Type 2 / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D014281:Trihexosylceramides / Q000378:metabolism*; D000519:alpha-Galactosidase / Q000378:metabolism; D000099183:Sortilin
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D015415:Biomarkers; D005964:Glucosyltransferases; D006028:Glycosphingolipids; D037181:Lectins, C-Type; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D017527:Receptor, IGF Type 2; D011956:Receptors, Cell Surface; D014281:Trihexosylceramides; D000519:alpha-Galactosidase; D000099183:Sortilin; C018549:globotriaosylceramide; C023246:ceramide glucosyltransferase