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Kidney Week

Abstract: TH-OR015

Single-Nuclear Transcriptomics Reveal Distinct Profibrotic Genes in Proximal Tubular and Endothelial Cells in a Novel Swine Model of CKD

Session Information

Category: CKD (Non-Dialysis)

  • 2303 CKD (Non-Dialysis): Mechanisms

Authors

  • Chade, Alejandro R., University of Missouri, Columbia, Missouri, United States
  • Eirin, Alfonso, Mayo Clinic Minnesota, Rochester, Minnesota, United States
Background

Renal fibrosis is the final stage of chronic kidney disease (CKD), irrespective of the etiology, and strategies to offset the progression to end-stage kidney disease are needed. We took advantage of a novel translational model of CKD in swine and single-nucleus RNA sequencing (snRNA-seq) to characterize the renal cell-specific transcriptomic landscape and identify pro-fibrotic genes that could serve as potential therapeutic targets.

Methods

Kidneys from normal and CKD pigs (n=3/group) were harvested, nuclei isolated, libraries prepared, and snRNA-seq performed. UMAP plots were generated with Seurat (v4.4.0) using default parameters. The expression profile of marker genes across single-nuclei clusters was visualized to define renal cell types and ranked based on the number of differentially expressed genes (DEGs). Endothelial and proximal tubular cells were the top cell types, subjected to subsequent analysis, and DEGs were sorted based on their involvement in kidney fibrosis.

Results

Thirty clusters were identified, filtered by canonical gene markers, and revealed 18 renal cell types (Fig. 1A-B). Endothelial and proximal tubular cells showed the highest number of pro-fibrotic DEGs (Fig. 1C-1D). LAMA3, upregulated in CKD endothelial cells, was selected for mechanistic investigation. LAMA3 partakes in ECM accumulation and the decline of renal function with aging but has not been linked to CKD. Notably, targeted silencing of LAMA3 (siRNA) ameliorated TGF-beta immunoreactivity, suggesting attenuated fibrogenic signaling (Fig. 1E).

Conclusion

TThe renal single-nuclear transcriptomic landscape of a preclinical model of CKD reveals endothelial and proximal tubular cells with the most significant number of pro-fibrotic DEGs. The candidacy of LAMA3 as a target to ameliorate pro-fibrotic activity could pave the way for new strategies to target fibrosis and slow the progression of CKD.

Funding

  • NIDDK Support

Digital Object Identifier (DOI)