Abstract: FR-PO803
Deciphering Antigen-Specific Glomerular Injury Mechanisms in Membranous Nephropathy via Single-Cell RNA Sequencing (scRNA-seq) and a Glomerulus-on-a-Chip Model
Session Information
- Glomerular Diseases: Inflammation and Immunology
October 25, 2024 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology
Authors
- Da Sacco, Stefano, Children's Hospital Los Angeles, Los Angeles, California, United States
- Perin, Laura, Children's Hospital Los Angeles, Los Angeles, California, United States
- Zhang, Qi, Children's Hospital Los Angeles, Los Angeles, California, United States
- Wetzels, Jack F., Radboud Universiteit, Nijmegen, Gelderland, Netherlands
- De Filippo, Roger E., Children's Hospital Los Angeles, Los Angeles, California, United States
- Cravedi, Paolo, Icahn School of Medicine at Mount Sinai, New York, New York, United States
Background
Primary membranous nephropathy (MN) is a prevalent cause of adult nephrotic syndrome, caused by anti-podocyte antibodies targeting proteins like PLA2R1 and THSD7A in the glomerular subepithelial space. This study explores the effects of these antibodies on glomerular cells using single-cell transcriptomics within a human glomerulus-on-a-chip (GOAC) model.
Methods
Human primary podocytes and glomerular endothelial cells were cultured within GOACs and subjected to sera containing anti-PLA2R or anti-THSD7A antibodies for a duration of 72 hours. Sera from healthy donors served as controls. Albumin permeability assays confirmed cellular injury, and single-cell RNA sequencing (scRNA-seq) was performed at a depth of 80,000 reads per cell, with subsequent detailed transcriptomic evaluations, followed by comprehensive transcriptomic analysis.
Results
Anti-PLA2R and anti-THSD7A exposure induced albumin leakage in GOACs, signaling injury. scRNA-seq revealed a pronounced activation of the complement pathway. For example, in podocytes, antigen-specific injury-related pathways such as EIF2 and mTOR signaling, and PTEN in glomerular endothelial cells, exhibited differential responses to anti-PLA2R sera, as opposed to anti-THSD7A or healthy sera, suggesting that concurrent early injury mechanisms drive podocyte dysregulation together with the complement cascade.
Conclusion
The study validates the application of scRNA-seq for the analysis of human cells in GOACs exposed to MN patient-derived sera. This methodological synergy offers insights into the pathogenic mechanisms at play in MN, providing a foundation for the identification of new therapeutic avenues for glomerular disorders.
Funding
- NIDDK Support