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

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 degrees C [24 hours], -20 degrees C, and -80 degrees 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 degrees C and -20 degrees C resulted in significantly lower miRNA yield compared to storage at -80 degrees C (mean log2fold changes across miRNAs from -0.5 +/- 0.4 SD to -1.20 +/- 0.4 SD (4 degrees C versus -80 degrees C) and from -0.7 +/- 0.2 SD to -1.20 +/- 0.3 SD (-20 degrees C versus -80 degrees 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.