# Renal Single-Cell RNA Sequencing and Digital Cytometry in Dogs with X-Linked Hereditary Nephropathy

## TL;DR

This study uses advanced sequencing and digital cytometry to explore kidney disease in dogs, revealing cellular changes and potential treatment targets.

## Contribution

The study introduces the use of single-cell RNA sequencing and digital cytometry in a canine model of hereditary kidney disease.

## Key findings

- Differential gene expression was identified in pathways related to podocyte dysfunction and tubular inflammation.
- Machine learning-empowered digital cytometry revealed changes in cell type proportions across CKD stages.
- Single-cell methods uncovered cell-type specific pathways in proximal tubule cells affected by CKD.

## Abstract

Chronic kidney disease (CKD) is common in dogs, yet understanding the specific cellular changes in this condition remains challenging. In this study, we used single-cell RNA sequencing (scRNA-seq) to analyze kidney tissues from dogs with X-linked hereditary nephropathy (XLHN), a naturally occurring model of canine CKD. We identified differentially expressed genes in the pathways linked to podocyte dysfunction and tubular inflammation. These findings were validated using digital cytometry in additional dogs. The integration of these advanced techniques enhances our understanding of CKD progression in dogs and helps in identifying potential therapeutic targets.

Chronic kidney disease (CKD) significantly affects canine health, but the precise cellular mechanisms of this condition remain elusive. In this study, we used single-cell RNA sequencing (scRNA-seq) to profile renal cellular gene expression in a canine model of X-linked hereditary nephropathy (XLHN). Dogs with this condition exhibit juvenile-onset CKD similar to that seen in human Alport syndrome. Post-mortem renal cortical tissues from an affected male dog and a heterozygous female dog were processed to obtain single-cell suspensions. In total, we recovered up to 13,190 cells and identified 11 cell types, including major kidney cells and immune cells. Differential gene expression analysis comparing the affected male and heterozygous female dogs identified cell-type specific pathways that differed in a subpopulation of proximal tubule cells. These pathways included the integrin signaling pathway and the pathway for inflammation mediated by chemokine and cytokine signaling. Additionally, using machine learning-empowered digital cytometry, we deconvolved bulk mRNA-seq data from a previous canine study, revealing changes in cell type proportions across CKD stages. These results underline the utility of single-cell methodologies and digital cytometry in veterinary nephrology.