Single-Cell RNA Sequencing Delineates Renal Anti-Fibrotic Mechanisms Mediated by TRPC6 Inhibition

Area of Science:

• Nephrology
• Molecular Biology
• Immunology

Background:

• Chronic kidney disease (CKD) involves persistent inflammation and fibrosis, often leading to end-stage renal disease.
• Transient Receptor Potential Canonical 6 (TRPC6) channels are implicated in tubular injury and renal fibrosis.

Purpose of the Study:

• To define renoprotective cell compositions and transcriptional programs in response to TRPC6 inhibition.
• To explore the translational relevance of TRPC6 inhibition in human CKD.

Main Methods:

• Single-cell RNA sequencing (scRNA-Seq) of murine models (UUO and 2m post-I/R) treated with a selective TRPC6 inhibitor (SH045).
• scRNA-Seq analysis of human CKD kidney samples.
• Immunofluorescence and Western blot analysis to validate findings at the protein level.

Main Results:

• TRPC6 inhibition mitigated tubular injury and fibrosis in murine models.
• Global transcriptional shifts observed in CKD, including diverse inflammatory and endothelial cells.
• A novel endothelial cell subpopulation, ECRIN, was identified, regulating VEGF and GAS signaling.
• TRPC6 inhibition activated a Prnp transcription factor network, alleviating renal fibrosis.

Conclusions:

• TRPC6 inhibition demonstrates therapeutic potential for CKD by modulating cellular and transcriptional pathways.
• Identification of ECRIN cells and the Prnp network offers new insights into CKD pathogenesis and treatment strategies.