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bioRxiv · 10.1101/2021.07.29.454229

A multi-omics study to investigate tacrolimus nephrotoxicity mechanisms

AOUAD, H.·FAUCHER, Q.·SAUVAGE, F.-L.·PINAULT, E.·BARROT, C.-C.·ARNION, H.·MARQUET, P.·ESSIG, M.
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Abstract

BackgroundTacrolimus, an immunosuppressive drug prescribed to a majority of transplanted patients is nephrotoxic, through still unclear mechanisms. This study aims to evaluate the impact of tacrolimus on a lineage of proximal tubular cells using a multi-omics approach. MethodsLLC-PK1 cells were exposed to 5 M of tacrolimus for 24h. Intracellular proteins and metabolites, and extracellular metabolites were extracted and analysed by LC-MS/MS. The transcriptional expression of the dysregulated proteins PCK-1, FBP1 and FBP2 was measured using RT-qPCR. ResultsIn our cell model, tacrolimus impacted different metabolic pathways including those of arginine (e.g., citrulline, ornithine) (p < 0.0001), amino acids (e.g., valine, isoleucine, aspartic acid) (p < 0.0001) and pyrimidine (p<0.01). In addition, it induced oxidative stress (p < 0.01) as shown by a decrease in total cell glutathione quantity. It impacted cell energy through an increase in Krebs cycle intermediates (e.g., citrate, aconitate, fumarate) (p < 0.01) and down-regulation of PCK-1 (p < 0.05) and FPB1 (p < 0.01), which are key enzymes in gluconeogenesis. Apart from glucose synthesis, gluconeogenesis is an important process in kidney mediated acid-base balance control. ConclusionThe variations found using this multi-omics approach clearly point towards a dysregulation of energy production in epithelial cells of the renal tubule, and potentially of their functions, that may be implicated in tacrolimus nephrotoxicity in the clinics.

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AOUAD, H., FAUCHER, Q., SAUVAGE, F.-L., PINAULT, E., BARROT, C.-C., ARNION, H., MARQUET, P., ESSIG, M.. 2021-07-30. A multi-omics study to investigate tacrolimus nephrotoxicity mechanisms. https://doi.org/10.1101/2021.07.29.454229

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