NU13 - Renal Single-cell RNA Sequencing in Dogs With a Naturally Occurring Progressive Chronic Kidney Disease

Abstract:

Background: Single-cell RNA sequencing (scRNA-seq) is a novel technology to profile gene expression on the cellular level. To date, the application of scRNA-seq in canine tissues is limited, including in chronic kidney disease.

Hypothesis/Objectives: We aimed to identify cell clusters in diseased canine kidney tissues and hypothesized that altered gene expression can be detected in advanced CKD.

Animals: One affected and one carrier dog from a research colony with X-linked hereditary nephropathy, a naturally occurring animal model for canine CKD.

Methods: Prospective study. One gram of fresh kidney cortex was collected after euthanasia for renal failure. Single-cell suspension was submitted for library preparation using the Chromium Single-Cell 3′ Kit and sequenced on an Illumina NovaSeq. Data were analyzed and annotated using CellRanger, CellMarker, and PanglaoDB. Differential expression analysis was performed in Seurat R-packages with the Wilcoxon Rank Sum test (adjusted P-value < 0.05).

Results: In total, we recovered 15,212 cells and identified 14 cell clusters in the kidney tissues, including major kidney cells (proximal and distal tubule cells, podocytes, mesangial cells, and collecting duct cells) and inflammatory cells. Comparing renal cells in the affected dog (rapid disease progression) to the carrier (slow disease progression), 133 differentially expressed genes were identified, such as MT2A (P < 4.65E-35) and APOC3 (P < 2.74E-68) that were involved in oxidative damage, fibrosis, and impeded vascular regeneration in human CKD.

Conclusions and clinical importance: We showed that scRNA-seq is suitable for discovering promising therapeutic targets in canine tissues and providing new insights into CKD pathogenesis.