Stability and profiling of urinary microRNAs in healthy cats and cats with pyelonephritis or other urological conditions

Abstract Background Specific biomarkers of pyelonephritis (PN) in cats are lacking. MicroRNAs (miRNAs) have diagnostic potential in human nephropathies. Objectives To investigate the presence/stability of miRNAs in whole urine of cats and the discriminatory potential of selected urinary miRNAs for PN in cats. Animals Twelve healthy cats, 5 cats with PN, and 13 cats with chronic kidney disease (n = 5), subclinical bacteriuria (n = 3), and ureteral obstructions (n = 5) recruited from 2 companion animal hospitals. Methods Prospective case‐control study. Expression profiles of 24 miRNAs were performed by quantitative PCR (qPCR). Effect of storage temperature (4°C [24 hours], −20°C, and −80°C) was determined for a subset of miRNAs in healthy cats. Results Urinary miR‐4286, miR‐30c, miR‐204, miR4454, miR‐21, miR‐16, miR‐191, and miR‐30a were detected. For the majority of miRNAs tested, storage at 4°C and −20°C resulted in significantly lower miRNA yield compared to storage at −80°C (mean log2fold changes across miRNAs from −0.5  ± 0.4 SD to −1.20 ± 0.4 SD (4°C versus −80°C) and from −0.7 ± 0.2 SD to −1.20 ± 0.3 SD (−20°C versus −80°C)). Cats with PN had significantly upregulated miR‐16 with a mean log2fold change of 1.0 ± 0.4 SD, compared with controls (−0.1 ± 0.2, P = .01) and other urological conditions (0.6 ± 0.3, P = .04). Conclusions Upregulation of miR16 might be PN‐specific, pathogen‐specific (Escherichia coli), or both.