Altered status of glutathione and its metabolites in cystinotic cells

• Published in: Nephrology, dialysis, transplantation dialysis, transplantation, 2005-09, Vol.20 (9), p.1828-1832
• Main Authors: Levtchenko, Elena, de Graaf-Hess, Adriana, Wilmer, Martijn, van den Heuvel, Lambertus, Monnens, Leo, Blom, Henk
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Biological and medical sciences; Blood. Blood and plasma substitutes. Blood products. Blood cells. Blood typing. Plasmapheresis. Apheresis; Bone marrow, stem cells transplantation. Graft versus host reaction; Cells, Cultured; Child; Child, Preschool; Cornea - metabolism; Cystine - metabolism; cystinosis; Cystinosis - metabolism; Dipeptides - metabolism; Emergency and intensive care: renal failure. Dialysis management; Female; fibroblasts; Fibroblasts - metabolism; glutathione; Glutathione - blood; Glutathione - metabolism; glutathione disulfide; Glutathione Disulfide - metabolism; Humans; Intensive care medicine; Male; Medical sciences; Neutrophils - metabolism; Oxidative Phosphorylation; polymorphonuclear cells; Skin - cytology; Skin - metabolism; Skin - pathology; Transfusions. Complications. Transfusion reactions. Cell and gene therapy
• ISSN: 0931-0509; 1460-2385
• DOI: 10.1093/ndt/gfh932

Background. Cystinosis is an autosomal recessive disorder, caused by mutations of the lysosomal cystine carrier cystinosin, encoded by the CTNS gene (17p13). The concomitant intralysosomal cystine accumulation leads to multi-organ damage, with kidneys being the first affected. Altered mitochondrial oxidative phosphorylation has been demonstrated in animal proximal tubules loaded with cystine dimethyl ester, mimicking cystine accumulation in cystinosis, but has not been confirmed in cells of patients with cystinosis. Furthermore, the link between cystine accumulation and mitochondrial damage is also missing. We hypothesized that cytosolic cysteine deficiency resulting in intracellular glutathione (GSH) shortage might be involved in cellular dysfunction in cystinosis. Methods. Components of the γ-glutamyl cycle were measured in cultured skin fibroblasts (n = 9) and polymorphonuclear (PMN) leukocytes (n = 15) derived from patients with cystinosis and compared with the values in cultured fibroblasts (n = 9) and PMN cells (n = 18) of healthy controls. Results. Cystine content in cystinotic fibroblasts and PMN cells was significantly elevated compared with the controls, consistent with the lysosomal cystine accumulation in these cells. Although no reduction of total intracellular GSH content was found in cystinotic cells, it inversely correlated with cystine levels. Furthermore, GSH disulfide (GSSG) was elevated in cystinotic cells, resulting in an increased GSSG/total GSH (%) ratio. No relationship between intracellular cystine and GSH was found in control fibroblasts and PMN cells. Conclusion. An elevated GSSG/total GSH (%) ratio might indicate increased oxidative stress present in cystinotic cells. Inverse correlation between cystine accumulation and intracellular GSH content indicates that under stress conditions such as intensive energy demand or increased oxidative insult, cystinotic cells may be more prone to GSH depletion.