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Renal Single-Cell RNA Sequencing and Digital Cytometry in Dogs with X-Linked Hereditary Nephropathy
Candice P Chu1, Daniel Osorio2, Mary B Nabity1
1Department of Veterinary Pathobiology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX 77843, USA.
Animals : an Open Access Journal From MDPI
|July 29, 2025
Summary
Single-cell RNA sequencing revealed distinct cellular pathways in proximal tubule cells of dogs with X-linked hereditary nephropathy, a model for canine chronic kidney disease. This highlights new cellular mechanisms in kidney disease progression.
Area of Science:
- Veterinary Medicine
- Nephrology
- Genomics
Background:
- Chronic kidney disease (CKD) poses a significant health burden in dogs.
- The specific cellular mechanisms underlying canine CKD, particularly in genetic forms, are not well understood.
- X-linked hereditary nephropathy (XLHN) in dogs serves as a relevant model for human Alport syndrome, offering insights into juvenile-onset CKD.
Purpose of the Study:
- To investigate the cellular and molecular mechanisms of CKD in a canine model using single-cell RNA sequencing (scRNA-seq).
- To identify cell-type specific gene expression differences in canine X-linked hereditary nephropathy (XLHN).
- To explore the utility of advanced computational methods like digital cytometry in analyzing canine kidney disease.
Main Methods:
- Single-cell RNA sequencing (scRNA-seq) was performed on renal cortical tissues from an affected male and a heterozygous female dog with XLHN.
- Identification and annotation of major kidney and immune cell types were conducted.
- Differential gene expression analysis was used to compare cell populations between the affected and unaffected dogs.
- Machine learning-powered digital cytometry was employed to deconvolute bulk mRNA sequencing data from a previous study.
Main Results:
- Eleven distinct cell types were identified from approximately 13,190 recovered cells.
- A subpopulation of proximal tubule cells in the affected dog showed differential expression in pathways related to integrin signaling and chemokine/cytokine-mediated inflammation.
- Digital cytometry analysis revealed alterations in cell type proportions across different stages of canine CKD.
Conclusions:
- Single-cell RNA sequencing provides a high-resolution view of cellular heterogeneity in canine kidney disease.
- Specific signaling and inflammatory pathways are dysregulated in proximal tubule cells of dogs with XLHN.
- scRNA-seq and digital cytometry are valuable tools for advancing research in veterinary nephrology and understanding CKD pathogenesis.
