Pd63-10 Single Cell Analysis Reveals the Effect of Anti-Pla2r and Anti Thsd7a Sera on Human Glomerular Cells

You have accessJournal of UrologyInfections/Inflammation/Cystic Disease of the Genitourinary Tract: Kidney & Bladder II (PD63)1 May 2024PD63-10 SINGLE CELL ANALYSIS REVEALS THE EFFECT OF ANTI-PLA2R AND ANTI THSD7A SERA ON HUMAN GLOMERULAR CELLS Qi Zhang, Jack Wetzels, Paolo Cravedi, Laura Perin, and Stefano Da Sacco Qi ZhangQi Zhang , Jack WetzelsJack Wetzels , Paolo CravediPaolo Cravedi , Laura PerinLaura Perin , and Stefano Da SaccoStefano Da Sacco View All Author Informationhttps://doi.org/10.1097/01.JU.0001009384.23104.ca.10AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Primary membranous nephropathy (MN) is a leading cause of nephrotic syndrome in adults due to the deposition of anti-podocyte antibodies in the glomerular subepithelial space. Several podocyte proteins have been identified as targets of these autoantibodies, with PLA2R1 and THSD7A making up about 80-85% of the total. We investigated the specific effect of anti-PLA2R and anti-THSD7A on glomerular cells using single cell transcriptomics on a human glomerulus-on-a-chip (GOAC) system. METHODS: GOACs were generated using human primary podocytes and glomerular endothelial cells and cultured with serum containing anti-PLA2R+ or anti-THSD7A+ antibodies for 72 hours. Sera from healthy individuals were used as a control. Three samples from different individuals were used for each group. Albumin leakage assay was performed on GOAC to confirm injury. Single cell RNA-seq analysis (scRNA-seq) with a depth of 30K reads/cell was performed on isolate cells. Downstream analyses were done using UMAP, gene and pathway enrichment, and intra- and inter-cluster comparative transcriptomics. RESULTS: Exposure to anti-PLA2R and anti-THSD7A sera from MN patients confirmed injury on the GOAC as shown by onset of albumin leakage. scRNAseq analysis showed robust activation of the complement pathway in both cohorts, as expected. Preliminary analysis also showed different activation of injury pathways in both human podocytes and human glomerular endothelial cells. CONCLUSIONS: In conclusion, we have successfully performed scRNAseq data on human cells obtained from GOACs exposed to patients' sera. This combination of technologies will help us unravel the glomerular mechanisms of injury in MN and provide potential new targets for the treatment of nephropathies and other glomerular diseases. Source of Funding: NIH NIDDK © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e1296 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Qi Zhang More articles by this author Jack Wetzels More articles by this author Paolo Cravedi More articles by this author Laura Perin More articles by this author Stefano Da Sacco More articles by this author Expand All Advertisement PDF downloadLoading ...