Mp69-01 Glomerular Endothelial Dysfunction Contributes to Disease Progression in Alport Syndrome

You have accessJournal of UrologyInfections/Inflammation/Cystic Disease of the Genitourinary Tract: Kidney & Bladder I (MP69)1 May 2024MP69-01 GLOMERULAR ENDOTHELIAL DYSFUNCTION CONTRIBUTES TO DISEASE PROGRESSION IN ALPORT SYNDROME Sargis Sedrakyan, Hasmik Soloyan, Matthew Thornton, Geremy Clair, Janielle Cuala, Qi Zhang, Senta Georgia, Paolo Cravedi, Andrea Angeletti, Roger De Filippo, Stefano Da Sacco, and Laura Perin Sargis SedrakyanSargis Sedrakyan , Hasmik SoloyanHasmik Soloyan , Matthew ThorntonMatthew Thornton , Geremy ClairGeremy Clair , Janielle CualaJanielle Cuala , Qi ZhangQi Zhang , Senta GeorgiaSenta Georgia , Paolo CravediPaolo Cravedi , Andrea AngelettiAndrea Angeletti , Roger De FilippoRoger De Filippo , Stefano Da SaccoStefano Da Sacco , and Laura PerinLaura Perin View All Author Informationhttps://doi.org/10.1097/01.JU.0001008892.86171.de.01AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Chronic kidney disease (CKD) affects more than 10% of the population worldwide, and our understanding of the mechanisms in many forms of CKD are poorly understood. Alport syndrome (AS), is a form of CKD caused by mutations in the collagen IVα3, α4, or α5 gene. Even though injury to glomerular endothelial cells (GEC) in AS is well established, the role of GEC in Alport progression has not been elucidated. Here, we describe the role of mitochondria and lipid metabolism in GEC injury in an animal model of AS, and the potential of using amniotic fluid stem cell (AFSC) derived extracellular vesicles, (EVs) as a rescue strategy to restore glomerular homeostasis. METHODS: The phasor approach to fluorescent lifetime imaging microscopy (FLIM) was applied to evaluate the mitochondria and metabolic changes in GEC of AS vs WT mice. GEC isolated by FACS from Tek-tdT reporter AS and WT mice were compared by bulk RNA-seq, lipidomic and flow cytometric analysis. In vitro, silencing experiments on primary human GEC were performed to study the role of fatty acid synthase (FASN) in mitochondrial dysfunction and GEC damage. FASN-carrying AFSC-EVs and control nanoparticles were applied both in vitro and in vivo to restore lipid homeostasis in GEC. RESULTS: FLIM studies showed strong correlation between the metabolic state of GEC and the age or severity of disease in AS mice. RNA-seq analysis revealed changes in the pathways associated with lipid metabolism and mitochondria function. Mitochondrial dysfunction was confirmed using flow cytometric analysis of the MitoTracker signal in tdTomato expressing GEC. Lipidome analysis revealed high abundance of triglycerides in AS GEC. We confirmed accumulation of lipid droplets in the glomeruli of AS mice and in FASN KO human primary GEC, in vitro. These results suggest potential mitochondrial dysfunction in GEC. AFSC derived EV treatment restored lipid homeostasis in GEC, both in vitro and in vivo. CONCLUSIONS: We report for the first-time mitochondrial dysfunction in Alport GEC, and the ability of AFSC-derived EVs to rescue this phenotype. Better understanding of the metabolic changes in AS GEC could lead to the development of targeted new therapies for other forms of CKD. Source of Funding: California Institute for Regenerative MedicineNIDDK, National Institute of Health © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e1118 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Sargis Sedrakyan More articles by this author Hasmik Soloyan More articles by this author Matthew Thornton More articles by this author Geremy Clair More articles by this author Janielle Cuala More articles by this author Qi Zhang More articles by this author Senta Georgia More articles by this author Paolo Cravedi More articles by this author Andrea Angeletti More articles by this author Roger De Filippo More articles by this author Stefano Da Sacco More articles by this author Laura Perin More articles by this author Expand All Advertisement PDF downloadLoading ...