This study systematically analyzed the transcriptomic and cell communication network changes induced by the TRPC6 inhibitor SH045 in a mouse model of renal fibrosis using single-cell RNA sequencing technology, revealing the anti-inflammatory and anti-fibrotic functions of ECRIN endothelial cells and F2 fibroblasts, and providing a novel therapeutic target for chronic kidney disease.
Literature Overview
This paper, 'Single-Cell RNA Sequencing Delineates Renal Anti-Fibrotic Mechanisms Mediated by TRPC6 Inhibition', published in Advanced Science, reviews TRPC6 channel protein's critical regulatory role in renal fibrosis. By employing scRNA-Seq and spatial transcriptomics, the study comprehensively characterizes SH045's impact on cellular composition and inflammatory microenvironment in mouse UUO and 2m post-I/R models, and validates the existence of these cell subpopulations in CKD patient samples, providing theoretical foundations for TRPC6 as a therapeutic target.
Background Knowledge
Chronic kidney disease (CKD) is a globally prevalent disease with no effective cure, characterized by tubulointerstitial fibrosis and persistent low-grade inflammation. TRPC6 channel protein has emerged as a research hotspot for its regulatory role in fibrosis, with existing studies confirming its involvement in cardiac and renal fibrosis. However, its cell-type-specific mechanisms in kidneys remain unclear. This study systematically characterized SH045's regulatory network on cellular composition and signaling pathways in renal fibrosis models using scRNA-Seq and spatial transcriptomics. A novel endothelial cell subpopulation (ECRIN) was identified with its role in VEGF and GAS signaling, while F2 fibroblasts' anti-fibrotic mechanisms through Prnp transcription factor network were revealed. These findings expand our understanding of renal fibrosis cellular heterogeneity and provide molecular evidence for developing TRPC6-targeted therapies.
Research Methods and Experiments
The research team administered daily injections of SH045 or vehicle to mouse UUO and 2m post-I/R models. Single-cell RNA sequencing was performed using microfluidic chips, combined with spatial transcriptomics to characterize cell-type-specific gene expression changes. Key protein regulation (e.g., PrP, CD31, Ki67) was validated through immunofluorescence and Western blot. Pseudotime analysis and differentiation trajectories were conducted using Milo and Monocle3 algorithms to investigate signaling pathway activity dynamics. Cellular communication network remodeling was analyzed with CellChat and NCEM models, identifying the central roles of ECRIN endothelial cells and F2 fibroblasts in VEGF, GAS, and JAK-STAT signaling pathways.
Key Conclusions and Perspectives
Research Significance and Prospects
This study pioneers the single-cell characterization of TRPC6's cell-type-specific mechanisms in renal fibrosis, providing transcriptomic and signaling pathway evidence for developing anti-fibrotic therapies targeting TRPC6. Future studies should explore functional evolution of ECRIN and F2 fibroblasts across fibrosis stages and their conservation in other organ fibrosis. Cross-species validation using human CKD cohorts can assess TRPC6 inhibition's translational potential for clinical applications.
Conclusion
By integrating single-cell transcriptomics and spatial analysis, this study systematically elucidates the anti-inflammatory and anti-fibrotic mechanisms of TRPC6 inhibitor SH045 in renal fibrosis. The discovery of ECRIN endothelial cells and F2 fibroblasts introduces novel cytological perspectives for fibrosis research and establishes molecular foundations for TRPC6-targeted therapeutic strategies. These findings not only expand our understanding of CKD microenvironment dynamics but also identify potential targets for cell-type-specific treatments.
