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Expression analysis of lung miRNAs responding to ovine VM virus infection by RNAseq

ABSTRACT: MicroRNAs (miRNAs) are short endogenous, single-stranded, non-coding small RNA molecules of about 22 nucleotides in length. They regulate gene expression post-transcriptionally by silencing mRNA molecules and they regulate many physiological processes. Visna-Maedi virus (VMV) is a lentivirus that causes Visna-Maedi disease (VM) in sheep characterised by pneumonia, mastitis, arthritis and encephalitis; and affects cells of the monocyte/macrophage lineage. So far, there are no studies in the role of miRNAs in this viral disease. Using RNAseq technology and bioinformatics analysis the expression of miRNAs in different phases of the disease were studied. A total of 212 miRNAs were found, of which 46 were conserved sequences found for the first time in sheep and 12 were completely novel. Differential expression analysis showed changes in several miRNAs comparing uninfected and seropositive groups, but did not detect significant differences between seropositive asymptomatic and diseased sheep. The high increase in expression of oar-miR-21 agrees with the increase of the same miRNA detected in other viral diseases. In addition, the target prediction of dysregulated miRNAs revealed that they control genes involved in proliferation-related signalling pathways like PI3K-Akt, AMPK and ErbB.

ORGANISM(S): Ovis aries

PROVIDER: GSE115415 | GEO | 2018/12/19

REPOSITORIES: GEO

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Project description:Effective management and control of parasitic infections on farms depends on their early detection. Traditional serological diagnostic methods for Fasciola hepatica infection in livestock are specific and sensitive, but currently the earliest detection of the parasite only occurs at approximately three weeks post-infection. At this timepoint, parasites have already entered the liver and caused the tissue damage and immunopathology that results in reduced body weight and loss in productivity. Here, we investigated whether the differential abundance of micro(mi)miRNAs in sera of F. hepatica-infected sheep has potential as a tool for the early diagnosis of infection. Using miRNA sequencing analysis, we discovered specific profiles of sheep miRNAs at both the pre-hepatic and hepatic infection phases in comparison to non-infected sheep. In addition, six F. hepatica-derived miRNAs were specifically identified in sera from infected sheep. Thus, a panel of differentially expressed miRNAs comprising four sheep (miR-3231-3p; miR133-5p; 3957-5p; 1197-3p) and two parasite miRNAs (miR-124-3p; miR-Novel-11-5p) were selected as potential biomarkers. The expression of these candidates in sera samples from longitudinal sheep infection studies collected between 7 days and 23 weeks was quantified using RT-qPCR and compared to samples from age-matched non-infected sheep. We identified oar-miR-133-5p and oar-miR-3957-5p as promising biomarkers of fasciolosis, detecting infection as early as 7 days. The differential expression of the other selected miRNAs was not sufficient to diagnose infection; however, our analysis found that the most abundant forms of fhe-miR-124-3p in sera were sequence variants (IsomiRs) of the canonical miRNA, highlighting the critical importance of primer design for accurate diagnostic RT-qPCR. Accordingly, this investigative study suggests that certain miRNAs are biomarkers of F. hepatica infection and validates miRNA-based diagnostics for the detection of fasciolosis in sheep.

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Expression analysis of lung miRNAs responding to ovine VM virus infection by RNAseq

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