Abstract: FR-PO942
Single Cell RNA-Seq Identifies Molecular Fingerprints of Endothelial Cell Subpopulations in Kidney
Session Information
- Development, Stem Cells, Regenerative Medicine - II
October 26, 2018 | Location: Exhibit Hall, San Diego Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 501 Development, Stem Cells, and Regenerative Medicine: Basic
Authors
- Zhou, Alex Xianghua, AstraZeneca Gothenburg, Mölndal, Sweden
- He, Liqun, Uppsala University, Uppsala, Sweden
- Betsholtz, Christer, Uppsala University, Uppsala, Sweden
- Laerkegaard Hansen, Pernille B., AstraZeneca Gothenburg, Mölndal, Sweden
Background
The kidney consists of functionally and anatomically discrete vascular plexus, and correspondingly distinct endothelial cell (EC) subpopulations. The precise targeting of renal vascular plexus in treatment requires a comprehensive understanding of the molecular characteristics of EC subpopulations. This study aims to distinguish renal EC subpopulations through dissection of their molecular fingerprints using single cell RNA-Seq (scRNA-Seq) analysis.
Methods
The BTBR mouse was perfused with magnetic Dynabeads that were subsequently trapped in glomeruli. The kidneys were enzymatically digested and glomeruli containing Dynabeads were removed. The cell suspension was further incubated with Pecam1 antibody and Pecam1+ single cells were FACS sorted onto a 384-well plate. Single EC cDNA library was generated and RNA-Seq was performed on Illumina HiSeq 2500. Unsupervised clustering of EC subpopulations was performed with BackSPIN analysis.
Results
Pecam1 antibody selection of renal cell suspension produced a rather purified EC population, with ~80% successfully sequenced single cells being ECs. The ECs were clustered into 3 major subpopulations: the big vessel ECs (~30% of total ECs), peritubular capillary ECs (~36%), and glomerular ECs (~34%). These subpopulations displayed distinct molecular fingerprints. In particular, the glomerular and peritubular ECs showed differential gene expression patterns in vascular endothelial growth factor receptors, angiopoietin receptors, adrenomedullin system, caveolae-associated proteins, endocytic transport proteins, etc. The recent whole kidney scRNA-Seq analysis (Park et al, Science 2018) provided us an overall picture of the renal single cell population but limited resolution in ECs (Pecam1+ ~1% of total cell population, peritubular ECs ~10% of EC population). In contrast, a dedicated EC scRNA-Seq may facilitate balancing various EC subpopulations in a single study and revealing in-depth definition of EC subpopulations in kidney.
Conclusion
The renal EC subpopulations display certain distinct gene expression patterns as revealed by scRNA-seq, which underscores the potential of selecting specific mechanisms for targeted vascular therapies in renal diseases.
Funding
- Commercial Support – AstraZeneca Gothenburg