Abstract: TH-OR080

Single-Cell Analysis of Progenitor Cell Dynamics and Lineage Specification of the Human Fetal Kidney

Session Information

Category: Developmental Biology and Inherited Kidney Diseases

  • 401 Developmental Biology

Authors

  • Cebrian Ligero, Cristina, University of Michigan Medical School, Ann Arbor, Michigan, United States
  • Menon, Rajasree, University of Michigan Medical School, Ann Arbor, Michigan, United States
  • Otto, Edgar A., University of Michigan Medical School, Ann Arbor, Michigan, United States
  • Kokoruda, Austin, University of Michigan Medical School, Ann Arbor, Michigan, United States
  • Zhou, Jian, Princeton University, Princeton, New Jersey, United States
  • Zhang, Zidong, Princeton University, Princeton, New Jersey, United States
  • Troyanskaya, Olga, Princeton University, Princeton, New Jersey, United States
  • Spence, Jason R., University of Michigan Medical School, Ann Arbor, Michigan, United States
  • Kretzler, Matthias, University of Michigan Medical School, Ann Arbor, Michigan, United States
Background

The study of animal models has identified a plethora of genes and pathways driving the repetitive and reciprocal interactions between the Ureteric Bud (UB) and the Metanephric Mesenchyme (MM) that give rise to the collecting system and the nephron pool. However, these expression patterns and how they drive differentiation have not been systematically characterized in the human kidney. We have used single-cell transcriptomics to study individual cell dynamics and characterize the expression profile of the human kidney.

Methods

Fetal kidneys (105 to 115 days of gestation) were dissociated to single cells and processed for DropSeq workflow as described by the McCarroll lab. Individual cells were identified by barcodes, and transcripts were tagged with Unique Molecular Identifiers. Paired-end RNASeq was performed on a HiSeq2500 platform. Bioinformatics analysis employed the Picard tools developed by the Broad Institute and unsupervised clustering algorithms were executed with the R package toolkit “Seurat”. RNA profiles were mapped into trajectories derived using a nonparametric ridge estimation statistical framework. Gene expression was confirmed by immunofluorescence on fetal kidneys.

Results

Single cell transcriptome analyses of 3,865 cells (fig.A) enabled the distinction of UB-4 and MM-1 progenitors as well as their intermediate and differentiated lineages including the mature collecting ducts-18, the renal vesicle and comma- and s-shaped bodies-2, immature-9 and mature podocytes-13, proximal tubules-6, Henle’s loop and distal tubules-8, as well as mesangium-5 and cortical-5 and medullary interstitium-10. Importantly, known as well as novel markers for these cell types were revealed in the analysis.

Conclusion

We have generated an accurate map of gene expression and lineage relationships (fig.B) in the human fetal kidney. These results confirm the expression of genes identified by studying animal models. New gene-expression patterns have also been identified that may help understand human renal development.

Funding

  • NIDDK Support