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Establishment and characterization of Fabry disease endothelial cells with an extended lifespan
Jin-Song Shen1, Xing-Li Meng, Raphael Schiffmann
1Developmental 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.
Insights
Researchers developed a novel endothelial cell line from Fabry disease patients. This extended-lifespan cell line aids in studying Fabry disease vasculopathy and developing new therapies.
Area of Science:
- Genetics and Molecular Biology
- Cell Biology
- Rare Diseases
Background:
- Fabry disease is a genetic disorder causing alpha-galactosidase A deficiency.
- This leads to globotriaosylceramide accumulation, particularly in vascular endothelial cells, causing vasculopathy.
- Studying Fabry disease vasculopathy is challenging due to the limited lifespan of primary endothelial cells.
Purpose of the Study:
- To generate a long-lived endothelial cell line from a Fabry disease patient.
- To establish a reliable in vitro model for studying Fabry disease pathogenesis.
- To facilitate research into novel therapeutic strategies for Fabry disease.
Main Methods:
- Generated an endothelial cell line from a Fabry hemizygote by introducing the human telomerase reverse transcriptase gene.
- Characterized the cell line for endothelial markers (von Willebrand factor, CD31, CD34, eNOS) and functional properties.
- Assessed alpha-galactosidase A activity and globotriaosylceramide accumulation in the cell line.
Main Results:
- The generated cell line exhibited a significantly extended lifespan compared to primary cells.
- The cells maintained key endothelial cell markers and functional characteristics.
- The cell line demonstrated reduced alpha-galactosidase A activity and globotriaosylceramide accumulation, characteristic of Fabry disease.
Conclusions:
- A novel, extended-lifespan endothelial cell line from a Fabry disease patient has been successfully created.
- This cell line serves as a valuable in vitro model for investigating Fabry disease vasculopathy.
- The model will aid in understanding disease mechanisms and developing targeted therapies.
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.
