APOL1 polymorphism modulates sphingolipid profile of human podocytes
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ORIGINAL ARTICLE
APOL1 polymorphism modulates sphingolipid profile of human podocytes Manuela Valsecchi 1 & Valentina Cazzetta 1,2 & Ferdinando Oriolo 2 & Xiqian Lan 3 & Rocco Piazza 4 & Moin A. Saleem 5 & Pravin C. Singhal 6 & Domenico Mavilio 1,2 & Joanna Mikulak 1,2 & Massimo Aureli 1 Received: 18 June 2020 / Revised: 19 August 2020 / Accepted: 2 September 2020 # The Author(s) 2020
Abstract Apolipoprotein L1 (APOL1) wild type (G0) plays a role in the metabolism of sphingolipids, glycosphingolipids, sphingomyelin and ceramide, which constitute bioactive components of the lipid rafts (DRM). We asked whether APOL1 variants (APOL1-Vs) G1 and G2 carry the potential to alter the metabolism of sphingolipids in human podocytes. The sphingolipid pattern in HPs overexpressing either APOL1G0 or APOL1-Vs was analysed by using a thin mono- and bi-dimensional layer chromatography, mass-spectrometry and metabolic labelling with [1-3H]sphingosine. HP G0 and G1/G2-Vs exhibit a comparable decrease in lactosylceramide and an increase in the globotriaosylceramide content. An analysis of the main glycohydrolases activity involved in glycosphingolipid catabolism showed an overall decrease in the activeness of the tested enzymes, irrespective of the type of APOL1-Vs expression. Similarly, the high throughput cell live-based assay showed a comparable increased action of the plasma membrane glycosphingolipid-glycohydrolases in living cells independent of the genetic APOL1 expression profile. Importantly, the most significative modification of the sphingolipid pattern induced by APOL1-Vs occurred in DRM resulted with a drastic reduction of radioactivity associated with sphingolipids. G1/G2-Vs present a decrease amount of globotriaosylceramide and globopentaosylceramide compared to G0. Additionally, ceramide at the DRM site and lactosylceramide in general, showed a greatest fall in G1/G2 in comparison with G0. Additionally, the levels of glucosylceramide decreased only in the DRM of human podocytes overexpressing G1/G2-Vs. These findings suggest that altered sphingolipidsprofiles may contribute to the deranged functionality of the plasma membrane in APOL1 risk milieu. Keywords APOL1 . APOL1 polymorphism . HIVAN . Sphingolipids . Podocytes . Lipid rafts . Plasma membrane Joanna Mikulak and Massimo Aureli contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10719-020-09944-w) contains supplementary material, which is available to authorized users. * Massimo Aureli [email protected] 1
Department of Medical Biotechnologies and Translational Medicine (BioMeTra), University of Milan, Milan, Italy
2
Unit of Clinical and Experimental Immunology, Humanitas Clinical and Research Center – IRCCS, Rozzano, MI, Italy
3
Key Laboratory for Aging and Regenerative Medicine, School of Pharmacy, Southwest Medical University, Luzhou, Sichuan, China
4
Department of Medicine and Surgery, University of Milan-Bicocca, Monza, Italy
5
Pediatric Academic Renal Unit, Universi
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