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Abstract: TH-PO1076

Single-Cell Transcriptome Profiling of the Mouse and Human Glomerulus

Session Information

Category: Glomerular Diseases

  • 1204 Podocyte Biology

Authors

  • He, Bing, Karolinska Institutet , Stockholm, Sweden
  • Chen, Ping, Karolinska Institutet , Stockholm, Sweden
  • Zambrano Sevilla, Sonia, Karolinska Institutet , Stockholm, Sweden
  • Dabaghie, Dina, Karolinska Institutet , Stockholm, Sweden
  • Guo, Jing, Karolinska Institutet , Stockholm, Sweden
  • Lal, Mark, AstraZeneca, Gothenburg, Sweden
  • Sandberg, Rickard, Karolinska Institutet , Stockholm, Sweden
  • Patrakka, Jaakko, Karolinska Institutet , Stockholm, Sweden
Background

Though the mouse is widely used to model human glomerular diseases, systematic transcriptome comparison of principal cell types forming the glomerular filtration barrier between two species is lacking. To address this question, we generated single cell RNA sequencing (scRNA-seq) libraries from mouse and human glomerular cells.

Methods

Mouse glomeruli were isolated from 8 wt adult C57BL/6J mice via magnetic bead perfusion. Human healthy glomeruli were purified from 8 donor kidney biopsies via a sieving procedure. Viable single cells of enriched glomeruli were unbiasedly sorted to 384-well plates and scRNA-seq was performed using the Smart-seq2 protocol. For data comparison between 2 species, we focused on podocytes, glomerular endothelial cells (GEC) and mesangial cells (MC).

Results

In total, 2416 mouse cells and 788 human cells passed quality control. Unsupervised clustering of these cells identified Nphs1+ podocytes, Kdr+ GECs, Pdgfrb+ MCs and other cell types such as tubular cells and immune cells. Interestingly, Cldn1+ mouse parietal epithelial cells were captured. Overall comparison showed more genes detected in podocytes than other two cell types. In podocytes, about 70 genes were identified as human-specific, of which half of them showed restricted expression in podocytes. Most human-specific genes have not been implicated in the podocyte function. However, important exceptions were detected, such as PLA2R1 encoding a major autoantigen of human membranous nephropathy, which was absent in mouse podocytes. On the other hand, only 5 genes were identified as mouse-specific. In GEC and MC, < 20 genes showed apparent human-specificity and only 3 genes mouse-specificity. Differential species-specific cell expression patterns for selected genes were validated by analyzing bulk RNA-seq data, qPCR and immunostaining.

Conclusion

Our results highlight differences between mouse and human glomerular molecular signatures that are essential to design and interpret translational studies.

Funding

  • Commercial Support