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Single cell G-protein coupled receptor profiling of activated kidney fibroblasts expressing transcription factor 21
Harmandeep Kaur1, Veera Ganesh Yerra1, Sri Nagarjun Batchu1
1Keenan Research Centre for Biomedical Science and Li Ka Shing Knowledge Institute, St. Michael's Hospital, Toronto, Ontario, Canada.
Background And Purpose:
Activated fibroblasts deposit fibrotic matrix in chronic kidney disease (CKD) and G-protein coupled receptors (GPCRs) are the most druggable therapeutic targets. Here, we set out to establish a transcriptional profile that identifies activated kidney fibroblasts and the GPCRs that they express.
Experimental Approach:
RNA sequencing and single cell qRT-PCR were performed on mouse kidneys after unilateral ureteral obstruction (UUO). Candidate expression was evaluated in mice with UUO or diabetes or injected with adriamycin or folic acid. Intervention studies were conducted in mice with diabetes or UUO. Correlative histology was performed in human kidney tissue.
Key Results:
Transcription factor 21 (Tcf21)+ cells that expressed 2 or 3 of Postn, Acta2 and Pdgfra were highly enriched for fibrogenic genes and were defined as activated kidney fibroblasts. Tcf21+ α-smooth muscle actin (α-SMA)+ interstitial cells accumulated in kidneys of mice with UUO or diabetes or injected with adriamycin or folic acid, whereas renin-angiotensin system blockade attenuated increases in Tcf21 in diabetic mice. Fifty-six GPCRs were up-regulated in single Tcf21+ kidney fibroblasts, the most up-regulated being Adgra2 and S1pr3. Adenosine receptors, Adora2a/2b, were up-regulated in Tcf21+ fibroblasts and the adenosine receptor antagonist, caffeine decreased Tcf21 upregulation and kidney fibrosis in UUO mice. TCF21, ADGRA2, S1PR3 and ADORA2A/2B were each detectable in α-SMA+ interstitial cells in human kidney samples.
Conclusion And Implications:
Tcf21 is a marker of kidney fibroblasts that are enriched for fibrogenic genes in CKD. Further analysis of the GPCRs expressed by these cells may identify new targets for treating CKD.
Linked Articles:
This article is part of a themed issue on Translational Advances in Fibrosis as a Therapeutic Target. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v180.22/issuetoc.
Insights
This study identifies Transcription factor 21 (Tcf21) as a marker for activated kidney fibroblasts in chronic kidney disease (CKD). Targeting specific G-protein coupled receptors (GPCRs) expressed by these cells, like adenosine receptors, may offer new therapeutic strategies for treating kidney fibrosis.
Area of Science:
- Nephrology
- Cell Biology
- Pharmacology
Background:
- Activated fibroblasts contribute to fibrotic matrix deposition in chronic kidney disease (CKD).
- G-protein coupled receptors (GPCRs) represent promising therapeutic targets for fibrotic diseases.
Purpose of the Study:
- To define a transcriptional profile of activated kidney fibroblasts.
- To identify GPCRs expressed by activated kidney fibroblasts for potential therapeutic targeting in CKD.
Main Methods:
- RNA sequencing and single-cell qRT-PCR on mouse kidneys following unilateral ureteral obstruction (UUO).
- Evaluation of candidate gene expression in various kidney injury models (UUO, diabetes, adriamycin, folic acid).
- Intervention studies in diabetic and UUO mouse models, with correlative human kidney tissue analysis.
Main Results:
- Transcription factor 21 (Tcf21)+ cells expressing specific fibrogenic markers were identified as activated kidney fibroblasts.
- Upregulation of 56 GPCRs, notably Adgra2 and S1pr3, was observed in Tcf21+ fibroblasts.
- Caffeine, an adenosine receptor antagonist, reduced Tcf21 upregulation and kidney fibrosis in UUO mice, with TCF21, ADGRA2, S1PR3, and ADORA2A/2B detected in human kidney samples.
Conclusions:
- Tcf21 serves as a reliable marker for kidney fibroblasts enriched in fibrogenic genes in CKD.
- Further investigation of GPCRs in these activated fibroblasts could unveil novel therapeutic targets for CKD treatment.
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