{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T13:03:37Z","timestamp":1774962217009,"version":"3.50.1"},"reference-count":35,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2013,7,30]],"date-time":"2013-07-30T00:00:00Z","timestamp":1375142400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Experimental Physiology"],"published-print":{"date-parts":[[2013,10]]},"abstract":"<jats:title>New findings<\/jats:title><jats:p><jats:list list-type=\"explicit-label\">\n<jats:list-item><jats:p><jats:bold>What is the central question of this study?<\/jats:bold><\/jats:p><jats:p>Are the molecular and cellular consequences of cystine dimethylester loading different from an alternative and more specific model of the cystinotic kidney proximal tubule epithelial cell, obtained by <jats:italic>CTNS<\/jats:italic> gene silencing.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p><jats:bold>What is the main finding and its importance?<\/jats:bold><\/jats:p><jats:p>Using a human\u2010derived kidney proximal tubular cell line, HK\u20102, we demonstrated that cystine dimethylester loading induces detrimental effects independent of lysosomal cystine accumulation. <jats:italic>CTNS<\/jats:italic> gene silencing, which resulted in comparable levels of cystine accumulation, evidence of oxidative stress and reduced ATP concentration but unaltered ATP generation capacity, represents a more useful model for investigating biochemical alterations in cystinosis.<\/jats:p><\/jats:list-item>\n<\/jats:list><\/jats:p><jats:p>Using the cystine dimethylester (CDME) loading technique to achieve elevated lysosomal cystine levels, ATP depletion has previously been postulated to be responsible for the renal dysfunction in cystinosis, a genetic disorder characterized by an excessive accumulation of cystine in the lysosomes. However, this is unlikely to be the sole factor responsible for the complexity of cell stress associated with cystinosis. Moreover, CDME has been shown to induce a direct toxic effect on mitochondrial ATP generation. Using a human\u2010derived proximal tubular epithelial cell line, we compared the effects of CDME loading with small interfering RNA\u2010mediated cystinosin, lysosomal cystine transporter (<jats:italic>CTNS<\/jats:italic>) gene silencing on glutathione redox status, reactive oxygen species levels, oxidative stress index, antioxidant enzyme activities and ATP generating capacity. The CDME\u2010loaded cells displayed increased total glutathione content, extensive superoxide depletion, augmented oxidative stress index, decreased catalase activity, normal superoxide dismutase activity and compromised ATP generation. In contrast, cells subjected to <jats:italic>CTNS<\/jats:italic> gene inhibition demonstrated decreased total glutathione content, increased superoxide levels, unaltered oxidative stress index, unaltered catalase activity, induction of superoxide dismutase activity and normal ATP generation. Our data indicate that many CDME\u2010induced effects are independent of lysosomal cystine accumulation, which further underscores the limited value of CDME loading for studying the pathogenesis of cystinosis. <jats:italic>CTNS<\/jats:italic> gene inhibition, which results in intracellular cystine accumulation, is a more realistic approach for investigating biochemical alterations in cystinosis.<\/jats:p>","DOI":"10.1113\/expphysiol.2013.073809","type":"journal-article","created":{"date-parts":[[2013,6,29]],"date-time":"2013-06-29T09:32:52Z","timestamp":1372498372000},"page":"1505-1517","source":"Crossref","is-referenced-by-count":13,"title":["Cystine dimethylester loading promotes oxidative stress and a reduction in ATP independent of lysosomal cystine accumulation in a human proximal tubular epithelial cell 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