Mapping cyclosporine-induced changes in protein secretion by renal cells using stable isotope labeling with amino acids in cell culture (SILAC).

Fabien Lamoureux 1, 2, 3 Louis Noël Gastinel 1 Elodie Mestre 1 Pierre Marquet 1, 2, * Marie Essig 1, 4
* Auteur correspondant
1 PIST - Pharmacologie des Immunosuppresseurs et de la Transplantation
2 Service de Pharmacologie, toxicologie et pharmacovigilance [CHU Limoges]
3 Service de Néphrologie [Rouen]
4 Service de Néphrologie, Dialyse, Transplantations [CHU Limoges]
Abstract : Nephrotoxicity is an adverse event that strongly limits the use of the immunosuppressant cyclosporine in solid organ transplantation and the precise molecular mechanisms underlying this toxicity remain unclear. MS-based proteomic analysis of the secretome of HEK-293 renal cells exposed to cyclosporine was performed to identify changes in protein secretion, as a first step to discover potential biomarkers of such nephrotoxicity. To detect and quantify the perturbed proteins in the culture medium we used SILAC and nano-scale liquid chromatography followed by MALDI-TOF/TOF mass spectrometry. Among 106 proteins identified, 80 were quantified in both forward/reverse SILAC experiments and quantitative proteomic analysis revealed altered levels of expression for 24 secreted proteins. These included the down-regulation of a number of extracellular matrix/cell adhesion components, and the up-regulation of secreted cyclophilins A and B, macrophage inhibition factor and phosphatidylethanolamine-binding protein 1. These changes in protein secretion were not prevented by co-incubation with the antioxidant N-acetylcysteine, suggesting that they were not triggered by cyclosporine-induced oxidative stress. The results from the present study provide important new knowledge to gain insights into the molecular mechanisms of cyclosporine-related toxicity. Some of the proteins identified here should be tested as potential biomarkers of cyclosporine nephrotoxicity in subsequent clinical studies.
Type de document :
Article dans une revue
Journal of Proteomics, Elsevier, 2012, 75 (12), pp.3674-87. 〈10.1016/j.jprot.2012.04.024〉
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http://www.hal.inserm.fr/inserm-00925556
Contributeur : Pierre Marquet <>
Soumis le : mercredi 8 janvier 2014 - 11:23:57
Dernière modification le : mardi 5 juin 2018 - 10:14:37

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Fabien Lamoureux, Louis Noël Gastinel, Elodie Mestre, Pierre Marquet, Marie Essig. Mapping cyclosporine-induced changes in protein secretion by renal cells using stable isotope labeling with amino acids in cell culture (SILAC).. Journal of Proteomics, Elsevier, 2012, 75 (12), pp.3674-87. 〈10.1016/j.jprot.2012.04.024〉. 〈inserm-00925556〉

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