Abstract: FR-PO404
Angiotensin II-Induced Cellular and Transcriptional Remodeling of Mouse Kidney Stroma
Session Information
- Hypertension and CVD: Basic
November 03, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Hypertension and CVD
- 1601 Hypertension and CVD: Basic
Authors
- Nelson, Jonathan W., Oregon Health & Science University, Portland, Oregon, United States
- Gurley, Susan B., Oregon Health & Science University, Portland, Oregon, United States
Background
The kidney stroma consists of perivascular cells that play multiple crucial roles. Recent data have shown that murine kidney stromal cells abundantly express the type 1a angiotensin receptor, however, how these cells respond to AngII treatment is poorly understood. Moreover, the cellular heterogeneity of the kidney stroma has been poorly characterized at the molecular level because they comprise a small percentage of kidney cells.
Methods
Pdgfrb-creERT2 mice were crossed with the INTACT mouse to create Pdgfrb-INTACT mice which express GFP connected to the nuclear envelope protein. Pdgfrb-INTACT mice were implanted with osmotic minipumps containing either saline (Veh) or Angiotensin II (AngII, 1000ng/kg/min) for 21 days. Nuclei were extracted from kidneys followed by fluorescence activated nuclei sorting (FANS) and microfluidic partitioning (10X Genomics). After sequencing the resulting cDNA libraries and deconvolution (Cell Ranger) the dataset was dimensionally reduced, integrated, and evaluated for differentially expressed genes (DEGs) with Seurat.
Results
16449 nuclei were sequenced (9644 from Veh; 6805 from AngII). Nuclei were enriched for expression of Pdgfrb and divided into 7 populations (Figure A): cortical fibroblasts, medullary fibroblasts, papillary fibroblasts, adaptive fibroblasts, contractile cells, mesangial cells, and cycling cells. Veh and AngII treated nuclei were present in all cell populations (Figure B), but not at the same proportion. Suggestive of AngII inducing myofibroblast remodeling, there was an increase in cycling cells (5 in Veh vs. 31 in AngII) with a concurrent decrease in contractile cells (1394 in Veh vs. 607 in AngII). While each population had a unique set of DEGs, one gene that was more abundant in AngII treated cells was Pde10a.
Conclusion
The mouse kidney stroma is transcriptionally diverse with significant cellular heterogeneity. AngII treatment alters both the cellular composition of the kidney stroma as well as the genes that are expressed within each cell population, highlighting the significant impact of AngII.
Targeted Single-Nucleus RNAsequencing of Kidney Stromal Cells from Mice Treated with Angiotensin II.
Funding
- NIDDK Support