Abstract: FR-PO365
Single-Nucleus RNA Sequencing Identifies Sex-Specific Proximal Tubular Cell Differentiation Pathways
Session Information
- Genetics, Development, Regeneration
November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Development‚ Stem Cells‚ and Regenerative Medicine
- 500 Development‚ Stem Cells‚ and Regenerative Medicine
Authors
- Lu, Yuehan, Cardiff University, Cardiff, Cardiff, United Kingdom
- Liao, Chia-Te, Taipei Medical University, Taipei, Taiwan
- Talabani, Bnar, Cardiff University, Cardiff, Cardiff, United Kingdom
- Smith, Tanya A., Cardiff University, Cardiff, Cardiff, United Kingdom
- Andrews, Robert, Cardiff University, Cardiff, Cardiff, United Kingdom
- Deshpande, Sumukh, Cardiff University, Cardiff, Cardiff, United Kingdom
- Bowen, Timothy, Cardiff University, Cardiff, Cardiff, United Kingdom
- Taylor, Philip, Cardiff University, Cardiff, Cardiff, United Kingdom
- Fraser, Donald, Cardiff University, Cardiff, Cardiff, United Kingdom
Group or Team Name
- Wales Kidney Research Unit
Background
Postpartum kidney growth is substantial but proliferation and differentiation pathways underpinning nephron elongation are not well defined. Here we performed sequential characterization of mouse kidney transcriptomics at the single cell level to address this.
Methods
Single nuclear RNA sequencing (snRNAseq) was performed on freshly harvested kidney tissue from male and female mice at age 1, 2, 4, 12 weeks of age (n=2, a total of 16 mice) using the 10x platform. Sequencing employed the Illumina Novaseq platform and downstream analysis was conducted in R using Seurat and associated packages.
Results
Unbiased clustering was performed on 68,775 nuclei. All expected cell types were identified. High levels of proliferation were evident at early time points in some (eg. Tubular) but not other (eg. Podocyte) clusters. Proliferation was especially evident in Proximal Tubular Cells (PTC’s) which are the most abundant cell type in the adult kidney. Uniquely when compared to other kidney cell types, PTC’s demonstrated sex-specific expression profiles at 4 and 12 weeks. Mapping of PTC differentiation pathways using techniques including trajectory and RNA Velocity analyses delineated increasing PTC specialization and sex-specific phenotype specification. Ligand-receptor analysis identified key cues.
Conclusion
Our single-cell transcriptomics data provide a library of cellular states observed during kidney growth. We have identified PTC differentiation pathways that lead to sex-specific tubular cell phenotypes.