Abstract: TH-OR092
Glomerular Endothelial Metabolic Reprogramming in Alport Syndrome: Aberrant Mitochondrial Oxidative Phosphorylation (OXPHOS) Activation and Extracellular Vesicle-Mediated Rescue
Session Information
- Shaping Pediatric Kidney Disease: Developmental Insights, Stem Cell Systems, Novel Biomarkers, and More
October 22, 2026 | Location: Room 603, Convention Center
Abstract Time: 04:40 PM - 04:50 PM
Category: Genetic Diseases of the Kidneys
- 1202 Genetic Diseases of the Kidneys: Non-Cystic (Complex and Non-Cystic Monogenic)
Authors
- Soloyan, Hasmik, Children's Hospital Los Angeles, Los Angeles, California, United States
- Cuala, Janielle M., University of Southern California, Los Angeles, California, United States
- Nelson, Jonathan W., University of Southern California Keck School of Medicine, Los Angeles, California, United States
- Clair, Geremy, Pacific Northwest National Laboratory, Richland, Washington, United States
- Merscher, Sandra, University of Miami Miller School of Medicine, Miami, Florida, United States
- Fornoni, Alessia, University of Miami Miller School of Medicine, Miami, Florida, United States
- Georgia, Senta K., Children's Hospital Los Angeles, Los Angeles, California, United States
- De Filippo, Roger E., Children's Hospital Los Angeles, Los Angeles, California, United States
- Perin, Laura, Children's Hospital Los Angeles, Los Angeles, California, United States
- Sedrakyan, Sargis, Children's Hospital Los Angeles, Los Angeles, California, United States
Background
Alport syndrome (AS) is driven by type IV collagen mutations leading to progressive kidney disease. While structural defects of the glomerular basement membrane and podocyte injury are well described, the role of glomerular endothelial cells (GECs) remains poorly defined. We previously showed that GEC injury precedes podocyte loss, suggesting a primary role in disease initiation. Here, we investigate whether lipid driven mitochondrial dysfunction in GECs contributes to AS pathobiology and whether these abnormalities can be reversed by human amniotic fluid stem cell-derived extracellular vesicles (hAFSC-EVs).
Methods
Using the Col4a5-/- model, we integrated single-cell RNA sequencing, proteomics, lipidomics, and fluorescence lifetime imaging microscopy (FLIM) to define GEC-specific metabolic alterations across disease stages. Mechanistic studies were performed in primary human GECs with FASN knockdown. hAFSC-EVs were evaluated in vitro and in vivo using single- and two-dose regimens.
Results
AS GECs showed early metabolic reprogramming with reduced FASN, increased CD36, and triglyceride accumulation, establishing a lipotoxic phenotype. Multi-omics revealed enrichment of lipid metabolism and oxidative phosphorylation pathways, with mitochondrial depletion and a shift from glycolysis to OXPHOS, reflected by increased bound NADH on FLIM. FASN deficiency recapitulated these changes in human GECs, promoting fatty acid uptake, mitochondrial dysfunction, lipid accumulation, and impaired barrier function, with up to a 44% increase in permeability, linking metabolic injury to endothelial dysfunction. hAFSC-EVs reversed these changes by restoring FASN, ACACA, and CD36, normalizing NADH balance, reducing lipid burden, and improving barrier integrity in vitro. In vivo, effects were dose-dependent: one dose had minimal impact, while two doses reprogrammed endothelial transcription, suppressed lipid oxidation, and restored NADH balance toward wild-type levels, consistent with recovery.
Conclusion
These findings identify GECs as central drivers in Alport pathobiology through lipid metabolic dysregulation and mitochondrial stress. hAFSC-EVs reverse these defects, restoring endothelial homeostasis and barrier integrity. Targeting GEC metabolism reframes AS as a multicellular metabolic disease and provides a mechanistically grounded therapeutic strategy.
Funding
- NIDDK Support