Fecal miRNA Profiling of Yorkshire Terrier Enteropathy

Abstract

MicroRNAs (miRNAs) are small non-coding RNAs involved in gene regulation and are potential biomarkers for several diseases, including canine enteropathies. While metabolite profiling and microbiome in canine enteropathies have been previously explored, data on miRNA expression remain limited. This study aimed to profile miRNA expression in Yorkshire Terrier canine enteropathy using Illumina sequencing and quantitative PCR (qPCR) to compare miRNA levels between sick and healthy dogs from fecal samples. Despite the hypothesis that disease-related alterations in miRNA levels would differentiate sick dogs from controls, no significant differences were observed between the groups in either sequencing or qPCR analyses. These findings suggest that miRNA profiles may not vary significantly in the context of Yorkshire Terrier enteropathy and indicate that other molecular or metabolomic markers may be more indicative of disease state. This study also indicates that fecal samples may not be an ideal sample type for miRNA profiling. This study contributes to the understanding of molecular signatures in canine enteropathies and provides a basis for further research into alternative biomarkers for diagnosis and monitoring.

Keywords: IBD; biomarker; canine IBD; chronic enteropathy; inflammatory bowel diseases; miRNA.

Figure 1

Normalized relative expression of target miRNAs using the comparative ΔΔCT method for primers P1–P34. miRNA expression levels were quantified from fecal samples collected from Yorkshire Terriers with and without enteropathy. Bars represent normalized data. No statistically significant differences were observed between sick and healthy dogs in either sequencing or qPCR analyses. The primers employed in this analysis were validated by the manufacturer (miRCURY™ microRNA, Qiagen [25]), ensuring high specificity and sensitivity for the selected miRNAs. Despite this, the absence of detectable differences could be attributed to several factors. One possibility is biological variability within the groups, where individual differences in miRNA expression may have masked subtle changes. Additionally, it is possible that the selected miRNAs are not directly involved in YTE pathophysiology, and the disease may instead involve other molecular pathways or regulatory mechanisms.