title: Establishment and characterization of Fabry disease endothelial cells with an extended lifespan.
abstract: Fabry disease is an inborn error of glycosphingolipid catabolism resulting from a deficiency of lysosomal enzyme alpha-galactosidase A. The major clinical manifestations of the disease, such as stroke, cardiac dysfunction, and renal impairment, are thought to be caused by vasculopathy due to progressive accumulation of globotriaosylceramide in vascular endothelial cells. The pathogenesis of the vasculopathy has not been elucidated. Since in vitro studies using primary endothelial cells are hampered by the limited lifespan of these cells, the availability of cultured endothelial cells with an extended lifespan is critical for the study of the vasculopathy of Fabry disease. We therefore generated an endothelial cell line from a Fabry hemizygote by introduction of human telomerase reverse transcriptase gene. The cell line has markedly extended lifespan compared to parental primary cells. The cells stably express many key markers of endothelial cells such as von Willebrand factor, CD31, CD34, and endothelial nitric oxide synthase (eNOS) and retain functional characteristics such as uptake of acetylated low-density lipoprotein, responsiveness to angiogenic growth factors, up-regulation of eNOS production upon extracellular stimuli, and formation of tube-like structures on Matrigel basement membrane matrix. The cells show significantly reduced activity of alpha-galactosidase A compared with primary endothelial cells from normal individuals and accumulate globotriaosylceramide in lysosomes. This cell line will provide a useful in vitro model of Fabry disease and will facilitate systematic studies to investigate pathogenic mechanisms and explore new therapeutic approaches for Fabry disease.
authors.affiliation: Developmental and Metabolic Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Building 10, Room 3D04, 9000 Rockville Pike, Bethesda, MD 20892-1260, USA.
authors.forename: Jin-Song
authors.initials: JS
authors.lastname: Shen
authors.forename: Xing-Li
authors.initials: XL
authors.lastname: Meng
authors.forename: Raphael
authors.initials: R
authors.lastname: Schiffmann
authors.forename: Roscoe O
authors.initials: RO
authors.lastname: Brady
authors.forename: Christine R
authors.initials: CR
authors.lastname: Kaneski
mesh_terms: D000107:Acetylation; D018952:Antigens, CD34 / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002455:Cell Division; D002461:Cell Line, Transformed; D004730:Endothelium, Vascular / Q000378:metabolism / Q000473:pathology*; D000795:Fabry Disease / Q000473:pathology*; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D016133:Polymerase Chain Reaction; D019098:Telomerase / Q000378:metabolism; D014281:Trihexosylceramides / Q000378:metabolism; D014841:von Willebrand Factor / Q000378:metabolism
chemical_list: D018952:Antigens, CD34; D015415:Biomarkers; D015815:Cell Adhesion Molecules; D008077:Lipoproteins, LDL; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D014281:Trihexosylceramides; D014841:von Willebrand Factor; C018549:globotriaosylceramide; D052250:Nitric Oxide Synthase Type III; C509186:TERT protein, human; D019098:Telomerase