Abstract
Background
Cystinosis is an autosomal recessive disease caused by intralysosomal cystine accumulation, treated with cysteamine. Recently, new adverse effects of cysteamine were reported. Skin biopsies showed microvascular proliferation (angioendotheliomatosis). To examine the mechanism of angioendotheliomatosis associated with cysteamine toxicity, we examined the effect of cysteamine on human dermal microvascular endothelial cells (HDMVEC).
Methods
After cysteamine exposure (range 0–3.0 mM) during 24 h, cell viability was measured using water soluble tetrazolium salt-1 (WST-1) in both control HDMVEC and fibroblasts. Cell proliferation and apoptosis rate were measured in HDMVEC by bromodeoxyuridine (BrdU) incorporation and caspase 3 and caspase 7 activity, respectively. Intracellular glutathione (GSH) was measured in HDMVEC after cysteamine exposure of 0, 0.1 or 1.0 mM. Medium and cysteamine were refreshed every 6 h to mimic the in vivo situation. Next, cell viability in HDMVEC was measured after 24 h of GSH exposure (range 0–10.0 mM).
Results
HDMVEC viability and proliferation increased after cysteamine exposure 0.03–3.0 mM (p < 0.01) and 0.03–1.0 mM (p = 0.01) respectively; cell viability in fibroblasts was not affected by incubation with cysteamine. Apoptosis remained unaffected by incubation with 0–1.0 mM cysteamine, 3.0 mM caused increased apoptosis. Intracellular GSH was significantly increased after incubation with cysteamine 0.1 mM (p = 0.02) and 1.0 mM (p < 0.01). HDMVEC viability increased after exposure to GSH 1.0–5.0 mM (p < 0.01).
Conclusion
Cysteamine concentrations, similar to those described in plasma of cystinosis patients, stimulate HDMVEC viability and proliferation and increase intracellular GSH content. We postulate that this mechanism might underlie angioendotheliomatosis induced by cysteamine.
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Acknowledgements
Elena Levtchenko is supported by the Fund for Scientific Research, Flanders (Belgium) (F.W.O. Vlaanderen) by a Fundamental Clinical Investigatorship (1801110 N). Martine Besouw was supported by grants of Cystinosis Research Foundation and Cystinosis Research Network and Cystinosis Foundation Ireland.
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This study was funded by the Cystinosis Research Foundation, Cystinosis Research Network, Cystinosis Foundation Ireland and the Fund for Scientific Research, Flanders, Belgium (FWO Vlaanderen). All authors confirm their independence from the sponsors and declare that the content of the article has not been influenced by the sponsor.
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Communicated by: Bruce A Barshop
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Besouw, M., van den Heuvel, L., van Eijsden, R. et al. Increased human dermal microvascular endothelial cell survival induced by cysteamine. J Inherit Metab Dis 36, 1073–1077 (2013). https://doi.org/10.1007/s10545-013-9588-0
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DOI: https://doi.org/10.1007/s10545-013-9588-0