Abstract: TH-OR061

Single-Cell Transcriptome Profiling of Glomeruli from Healthy and Nephritic Mice

Session Information

Category: Glomerular

  • 1001 Glomerular: Basic/Experimental Immunology and Inflammation

Authors

  • Chung, Jun-Jae, Genentech, South San Francisco, California, United States
  • Chen, Jasmine, Genentech, South San Francisco, California, United States
  • Goldstein, Leonard, Genentech, South San Francisco, California, United States
  • Modrusan, Zora, Genentech, South San Francisco, California, United States
  • Shaw, Andrey S., Genentech, South San Francisco, California, United States
Background

Disruption of glomerular homeostasis leads to filtration barrier damage and kidney disease. Determining the gene expression profiles of cells that comprise the glomerulus will facilitate understanding of the processes involved in kidney diseases. Single-cell RNA-seq analysis can provide information not attainable using bulk RNA-seq, such as cell subpopulations and heterogeneity of cellular responses to injury.

Methods

Single cell suspensions of glomerular cells were prepared via magnetic bead isolation followed by enzymatic digestion from healthy C57BL/6J mice or mice with anti-GBM glomerulonephritis. The individual transcriptome of thousands of cells in the mouse glomeruli were obtained using the droplet-based 10x Genomics ChromiumTM technology and Next-Generation Sequencing.

Results

Principle component analysis (PCA) of cells from healthy glomeruli clearly distinguished the three major cell types in the glomerulus (podocytes, mesangial cells, endothelial cells). Analysis of differentially expressed genes identified novel candidates for mesangial cell markers. After induction of anti-GBM glomerulonephritis, we detected accumulation of immune cells and significant changes in the gene expression patterns of glomerular cells. After resolution of the heterologous phase, global gene expression pattern of podocytes largely reverted back to its original state, whereas the transcriptome profile of mesangial cells and endothelial cells remained altered. We observed heterogeneous expression of CCL2 in the mesangium of a CCL2-reporter mouse, which was increased after induction of glomerulonephritis. CCL2 expression was enriched in mesangial cells that had higher levels of immediate early genes, suggesting these cells may act as sentinels during glomerular injury. Comparing the transcriptomes of each cell type from healthy and nephritic glomeruli reveal candidate genes for further functional studies.

Conclusion

Single-cell RNA-seq reveals previously undetected heterogeneity in the injury response of glomerular cells.