Single-Nucleus RNA Sequencing Analysis of Interferon-Exposed APOL1 High-Risk Transgenic Mouse Reveals Distinct Injury Patterns in Podocytes and Endothelial Cells
- Genetic Diseases: Glomerulopathies - II
November 04, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1202 Genetic Diseases of the Kidneys: Non-Cystic
- Latt, Khun Zaw, National Institutes of Health, Bethesda, Maryland, United States
- Yoshida, Teruhiko, National Institutes of Health, Bethesda, Maryland, United States
- Shrivastav, Shashi, National Institutes of Health, Bethesda, Maryland, United States
- Heymann, Jurgen, National Institutes of Health, Bethesda, Maryland, United States
- Zhao, Yongmei, National Institutes of Health, Bethesda, Maryland, United States
- Rosenberg, Avi Z., Johns Hopkins University, Baltimore, Maryland, United States
- Kopp, Jeffrey B., National Institutes of Health, Bethesda, Maryland, United States
APOL1 high-risk genetic variants, termed G1 and G2, compared to the common variant G0, cause focal segmental glomerulosclerosis (FSGS), collapsing glomerulopathy, and arterionephrosclerosis in individuals with sub-Saharan African ancestry. Gene expression profiles of glomerular cells in APOL1-mediated cell injury have not been studied at the single cell level. We performed single-nucleus RNA sequencing (snRNA-seq) of glomeruli from wild-type (WT) and bacterial artificial chromosome (BAC)/APOL1-G0 and -G1 transgenic mice following interferon-g administration.
BAC/APOL1 transgenic mice received a single dose of interferon-g, administered retro-orbitally, and were euthanized 24 hours later. Glomeruli were isolated and nuclei were captured for snRNA-seq. The snRNA-seq data from BAC/APOL1-G0, BAC/APOL1-G1 and wild-type mice were integrated and analyzed using Seurat4. Podocytes and glomerular endothelial cell clusters were identified by expression of known marker genes. Subclustering and pseudotime analysis were performed to identify cell subpopulations and their characteristic gene signatures.
The combined analysis of snRNA-seq datasets identified podocyte clusters expressing Nphs1 and glomerular endothelial cell (GEC) cluster expressing Ehd3 and Hecw2. APOL1 was expressed by podocytes and endothelial cells. Expression levels were similar between APOL1-G0 and APOL1-G1 expressing cells within each cluster. Podocytes and GECs showed similar gene expression patterns of early injury but distinct patterns with more severe injury. Genes specific to the podocyte severe injury cluster in BAC/APOL1 mice showed higher expression in human FSGS APOL1 high-risk urinary podocytes, compared to APOL1-low-risk podocytes.
Following interferon administration to BAC/APOL1 mice, we observed a unique gene expression profile in a APOL1-G1 risk allele mouse compared to a APOL1-G0 mouse. APOL1 was expressed in podocytes and GECs. However, the gene expression patterns of injury in these two cell types were different and may reflect different roles by which these cells contribute to glomerular pathology in APOL1 glomerular disease.
- NIDDK Support