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. 2020 Jan;34(1):166-175.
doi: 10.1111/jvim.15628. Epub 2019 Nov 13.

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

Lisbeth R Jessen  1 , Lise N Nielsen  1 , Ida N Kieler  1 , Rebecca Langhorn  1 , Bert J Reezigt  2 , Susanna Cirera  3
Affiliations

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

Lisbeth R Jessen et al. J Vet Intern Med. 2020 Jan.

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.

Keywords: Biomarker; CKD; UTI; feline; pathogen-directed microRNA expression; subclinical bacteriuria; ureteral obstruction.

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Conflict of interest statement

Authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Flow diagram of the study set‐up and miRNAs tested in the different parts of the study
Figure 2
Figure 2
A‐H: The effect of storage temperature (4°C, −20°C, and −80°C) on the yield of the miRNAs used in the clinical study; A, miR‐16; B, miR‐30a; C, miR‐204; D, miR‐4286; E, miR‐21; F, miR‐30c; G, miR‐4454; H, miR‐191—stability study part II. Data represent cDNA duplicates from 5 healthy cats. Colors represent miRNA from the same cat‐sample stored at 3 different temperatures. *Statistically significant
Figure 3
Figure 3
First (PC1) and second (PC2) principal component, from the principal component analysis including the miRNAs (miR‐16, miR‐21, miR‐30c, miR‐204, miR‐30a, miR‐4286, miR‐191, and miR‐4454) from the clinical study, colored by group (chronic kidney disease [CKD]), healthy controls (Controls), ureteral obstruction (UO), pyelonephritis (PN), and subclinical bacteriuria (SB), and the name of the cultured pathogen when relevant
Figure 4
Figure 4
A‐D: Log2 fold changes of the miRNAs A, miR‐16; B, miR‐30a; C, miR‐204; D, miR‐4286 from the clinical study divided by group: pyelonephritis (PN), healthy controls (Controls), and other urological conditions (Other). *Statistically significant
See this image and copyright information in PMC

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