SILAC-Based Proteomics of Primary Human Kidney Cells treated with Sex Hormones.

ABSTRACT: Male sex predispose to many kidney diseases. Considering that androgens exert deleterious effects in a variety of cell types within kidney, we hypothesized that dihydrotestosterone (DHT) would impair the biology of the renal tubular cell by inducing changes in the proteome. We employed stable isotope labeling with amino acids (SILAC) in an indirect spike-in fashion to accurately quantify the proteome in DHT- and 17?-estradiol (EST)-treated human proximal tubular epithelial cells (PTEC), in 4 different experiments. Of the 5043 quantified proteins, 76 were differentially regulated. Biological processes related to energy metabolism were significantly enriched among DHT-regulated proteins. SILAC ratios of 3 candidates representing glycolysis, N-acetylglucosamine metabolism and fatty acid ?-oxidation, namely glucose-6-phosphate isomerase (GPI), glucosamine-6-phosphate-N-acetyltransferase 1 (GNPNAT1) and mitochondrial trifunctional protein subunit alpha (HADHA), were verified in vitro. In vivo, renal GPI and HADHA protein expression was increased in males. Furthermore, male sex was associated to higher GPI, GNPNAT1 and HADHA protein expression in diabetic Akita mice. Functional group enrichment analysis revealed a link between our DHT-regulated proteins and glycosphingolipid metabolism within the male kidney. This is the most in depth quantitative proteomic study of human primary PTEC response to sex hormone treatment. In this study we open new perspectives on how to explore the molecular mechanisms responsible for the deleterious effects of androgens in the context of kidney disease, especially diabetic nephropathy.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Ana Konvalinka  

PROVIDER: MSV000080854 | MassIVE | Fri Mar 31 23:44:00 BST 2017

SECONDARY ACCESSION(S): PXD003811

REPOSITORIES: MassIVE