Abstract: SA-PO729
Single-Cell RNA Sequencing of Myofibroblasts to Identify Therapeutic Targets in CKD
Session Information
- CKD: Mechanisms - III
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: CKD (Non-Dialysis)
- 2103 CKD (Non-Dialysis): Mechanisms
Authors
- He, Liqun, Uppsala University, Uppsala, Sweden
- Betsholtz, Christer, Uppsala University, Uppsala, Sweden
- Jeansson, Marie, Karolinska Institutet, Huddinge, Sweden
Background
Renal tubulointerstitial fibrosis is predictive of progressive decline in kidney function, independent of underlying disease. It is characterized by an increase in activated cells, myofibroblasts, which produce and deposit extracellular matrix. This project aim at an in depth analysis of myofibroblasts to further understand the progression of fibrosis and to identify novel targets for development of therapeutics.
Methods
Mice with Pdgfra-GFP and Pdgfrb-GFP reporters were utilized to identify pericytes, fibroblasts and myofibroblasts. Mice were subjected to unilateral ureter obstruction (UUO) and analyzed at different time points after UUO. GFP positive cells from UUO and contralateral (CL) kidneys were enzymatically digested and sorted into 384-well plates by FACS for single cell RNA-seq using SmartSeq2. Cluster analysis was performed with BackSPIN and Pagoda2.
Results
Pdgfra-GFP and Pdgfrb-GFP positive cells were significantly increased after UUO and colocalized with markers of fibrosis (ASMA and vimentin). Analysis of the single cell transcriptomes from Pdgfra-GFP and Pdgfrb-GFP cells clustered CL cells and UUO cells separately. Several genes were exclusively expressed in cells from UUO kidneys, while some genes were seen in CL kidneys but lost in UUO kidneys.
Ongoing studies include trajectory analysis from the different time point as well as confirmation of results with immunohistochemistry and RNAScope.
Conclusion
Our study will give insight into the temporal changes of fibroblast and pericytes transcriptomes on the single cell level during fibrosis progression.
Funding
- Private Foundation Support