Department of Animal Medicine and Surgery, School of Veterinary Sciences, Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Catalonia, Spain.
Centre for Research in Agricultural Genomics (CRAG) CSIC-IRTA-UAB-UB, Campus UAB, Cerdanyola del Vallès, Catalonia, Spain.
PLoS One. 2022 Jul 20;17(7):e0270067. doi: 10.1371/journal.pone.0270067. eCollection 2022.
MicroRNAs (miRNAs) are short non-coding RNAs that regulate gene expression at the post-transcriptional level. miRNAs have been found in urine and have shown diagnostic potential in human nephropathies. Here, we aimed to characterize, for the first time, the feline urinary miRNAome and explore the use of urinary miRNA profiles as non-invasive biomarkers for feline pyelonephritis (PN). Thirty-eight cats were included in a prospective case-control study and classified in five groups: healthy Control cats (n = 11), cats with PN (n = 10), cats with subclinical bacteriuria or cystitis (SB/C, n = 5), cats with ureteral obstruction (n = 7) and cats with chronic kidney disease (n = 5). By small RNA sequencing we identified 212 miRNAs in cat urine, including annotated (n = 137) and putative novel (n = 75) miRNAs. The 15 most highly abundant urinary miRNAs accounted for nearly 71% of all detected miRNAs, most of which were previously identified in feline kidney. Ninety-nine differentially abundant (DA) miRNAs were identified when comparing Control cats to cats with urological conditions and 102 DA miRNAs when comparing PN to other urological conditions. Tissue clustering analysis revealed that the majority of urine samples clustered close to kidney, which confirm the likely cellular origin of the secreted urinary miRNAs. Relevant DA miRNAs were verified by quantitative real-time PCR (qPCR). Eighteen miRNAs discriminated Control cats from cats with a urological condition. Of those, seven miRNAs were DA by both RNAseq and qPCR methods between Control and PN cats (miR-125b-5p, miR-27a-3p, miR-21-5p, miR-27b-3p, miR-125a-5p, miR-17-5p and miR-23a-3p) or DA between Control and SB/C cats (miR-125b-5p). Six additional miRNAs (miR-30b-5p, miR-30c, miR-30e-5p, miR-27a-3p, miR-27b-39 and miR-222) relevant for discriminating PN from other urological conditions were identified by qPCR alone (n = 4) or by both methods (n = 2) (P<0.05). This panel of 13 miRNAs has potential as non-invasive urinary biomarkers for diagnostic of PN and other urological conditions in cats.
微小 RNA（miRNAs）是一种短的非编码 RNA，可在转录后水平调节基因表达。已经在尿液中发现了 miRNAs，并显示出在人类肾病中的诊断潜力。在这里，我们首次旨在描述猫的尿 miRNA 组，并探索尿 miRNA 谱作为非侵入性生物标志物用于猫肾盂肾炎（PN）。38 只猫被纳入前瞻性病例对照研究，并分为五组：健康对照组猫（n = 11）、PN 猫（n = 10）、亚临床菌尿或膀胱炎猫（SB/C，n = 5）、输尿管阻塞猫（n = 7）和慢性肾病猫（n = 5）。通过小 RNA 测序，我们在猫尿中鉴定出 212 个 miRNAs，包括注释（n = 137）和推定的新 miRNAs（n = 75）。15 个最丰富的尿 miRNA 占所有检测到的 miRNA 的近 71%，其中大部分以前在猫的肾脏中发现。当比较对照组猫与有泌尿系统疾病的猫时，有 99 个差异丰富（DA）miRNAs，当比较 PN 与其他泌尿系统疾病时，有 102 个 DA miRNAs。组织聚类分析表明，大多数尿液样本与肾脏聚类密切相关，这证实了分泌尿 miRNA 的可能细胞来源。通过定量实时 PCR（qPCR）验证了相关的 DA miRNAs。18 个 miRNAs 将对照组猫与有泌尿系统疾病的猫区分开来。其中，7 个 miRNAs 通过 RNAseq 和 qPCR 方法在对照组和 PN 猫之间存在差异（miR-125b-5p、miR-27a-3p、miR-21-5p、miR-27b-3p、miR-125a-5p、miR-17-5p 和 miR-23a-3p）或在对照组和 SB/C 猫之间存在差异（miR-125b-5p）。通过 qPCR 单独（n = 4）或两种方法（n = 2）鉴定出另外 6 个（miR-30b-5p、miR-30c、miR-30e-5p、miR-27a-3p、miR-27b-39 和 miR-222）与区分 PN 和其他泌尿系统疾病相关的 miRNAs（P<0.05）。这组 13 个 miRNAs 具有作为 PN 和其他猫泌尿系统疾病诊断的非侵入性尿生物标志物的潜力。
