Abstract
Dogs with X-linked hereditary nephropathy (XLHN) are an animal model for Alport syndrome in humans and progressive chronic kidney disease (CKD). Using mRNA sequencing (mRNA-seq), we have characterized the gene expression profile affecting the progression of XLHN; however, the microRNA (miRNA, miR) expression remains unknown. With small RNA-seq and quantitative RT-PCR (qRT-PCR), we used 3 small RNA-seq analysis tools (QIAGEN OmicSoft Studio, miRDeep2, and CPSS 2.0) to profile differentially expressed renal miRNAs, top-ranked miRNA target genes, and enriched biological processes and pathways in CKD progression. Twenty-three kidney biopsies were collected from 5 dogs with XLHN and 4 age-matched, unaffected littermates at 3 clinical time points (T1: onset of proteinuria, T2: onset of azotemia, and T3: advanced azotemia). We identified up to 23 differentially expressed miRNAs at each clinical time point. Five miRNAs (miR-21, miR-146b, miR-802, miR-142, miR-147) were consistently upregulated in affected dogs. We identified miR-186 and miR-26b as effective reference miRNAs for qRT-PCR. This study applied small RNA-seq to identify differentially expressed miRNAs that might regulate critical pathways contributing to CKD progression in dogs with XLHN.
Highlights
Dogs with X-linked hereditary nephropathy (XLHN) are an animal model for Alport syndrome in humans and progressive chronic kidney disease (CKD)
Using quantitative RT-PCR (qRT-PCR), we examined the expression of these 7 miRNAs using 10 XLHN affected canine kidney tissues sequenced in the current study and 2 canine kidney tissue controls used in a previous s tudy[3] so there were 2 samples from each group at each
Dogs with XLHN have been studied as an example of canine CKD and used as an animal model for human Alport syndrome
Summary
Dogs with X-linked hereditary nephropathy (XLHN) are an animal model for Alport syndrome in humans and progressive chronic kidney disease (CKD). This study applied small RNA-seq to identify differentially expressed miRNAs that might regulate critical pathways contributing to CKD progression in dogs with XLHN. We aimed to identify differentially expressed renal miRNAs that may contribute to CKD progression in dogs by regulating gene expression. We used small RNA-seq on RNA isolated from 23 canine kidney biopsies to characterize and compare miRNA expression in 5 dogs with XLHN and 4 controls at 3 clinical time points (T1: onset of proteinuria, T2: onset of azotemia, and T3: advanced azotemia). Based on the small RNA-seq data from 23 biopsies, CPSS 2.0 produced better genome mapping rate and identified more differentially expressed miRNAs. We conducted gene ontology (GO) and pathway analyses to characterize the miRNA targets among sample groups at specific time points. Our results deepen the understanding of the molecular mechanisms in Alport
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