Integrated Lung and Tracheal mRNA-Seq and miRNA-Seq Analysis of Dogs with an Avian-Like H5N1 Canine Influenza Virus Infection

Abstract

Avian-like H5N1 canine influenza virus (CIV) causes severe respiratory infections in dogs. However, the mechanism underlying H5N1 CIV infection in dogs is unknown. The present study aimed to identify differentially expressed miRNAs and mRNAs in the lungs and trachea in H5N1 CIV-infected dogs through a next-generation sequencing-based method. Eighteen 40-day-old beagles were inoculated intranasally with CIV, A/canine/01/Guangdong/2013 (H5N1) at a tissue culture infectious dose 50 (TCID50) of 106, and lung and tracheal tissues were harvested at 3 and 7 d post-inoculation. The tissues were processed for miRNA and mRNA analysis. By means of miRNA-gene expression integrative negative analysis, we found miRNA–mRNA pairs. Lung and trachea tissues showed 138 and 135 negative miRNA–mRNA pairs, respectively. One hundred and twenty negative miRNA–mRNA pairs were found between the different tissues. In particular, pathways including the influenza A pathway, chemokine signaling pathways, and the PI3K-Akt signaling pathway were significantly enriched in all groups in responses to virus infection. Furthermore, dysregulation of miRNA and mRNA expression was observed in the respiratory tract of H5N1 CIV-infected dogs and notably, TLR4 (miR-146), NF-κB (miR-34c) and CCL5 (miR-335), CCL10 (miR-8908-5p), and GNGT2 (miR-122) were found to play important roles in regulating pathways that resist virus infection. To our knowledge, the present study is the first to analyze miRNA and mRNA expression in H5N1 CIV-infected dogs; furthermore, the present findings provide insights into the molecular mechanisms underlying influenza virus infection.

The miRNA-targeted transcriptome of porcine alveolar macrophages upon infection with Porcine Reproductive and Respiratory Syndrome Virus

“…The absence of detectable phenotypes effects on macrophage function and levels of PRRSV infection cannot be solely accounted to the reduced sensitivity of miRNA mimics in PAMs. The miR-335-5p has been reported as one of the miRNAs down-regulated in PBMCs from HIV-1 and HIV-2 infected patients 58 and for having a role in the pathogenesis of influenza in the respiratory tract of dogs 59 . However, a recent atlas has shown that this miRNA locus is expressed at low levels in most tissues and cell types of the lung tract 60 .…”

Intestinal Expression of miR-130b, miR-410b, and miR-98a in Experimental Canine Echinococcosis by Stem-Loop RT-qPCR

“…We showed that cfa-miR-410 gene expression was upregulated in the intestinal cells of dogs infected experimentally with E. granulosus. In an avian-like H5N1 canine influenza virus model, cfa-miR-410 expression was decreased in the lung and tracheal tissues (34). Several studies on miR-410 suggested probable links with immune responses.…”

Interactions Among Expressed MicroRNAs and mRNAs in the Early Stages of Fowl Adenovirus Aerotype 4-Infected Leghorn Male Hepatocellular Cells

“…Each group was processed with three independent replicates. Total RNA was extracted from the cells using TRIzol (Ambion, Life Technologies, Carlsbad, CA, United States) according to the manual instructions (Fu et al, 2018). Subsequently, total RNA was qualified and quantified using a Nano Drop and Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, United States).…”

“…Twelve total RNA samples were obtained to generate the cDNA libraries for each sample according to the manufacturer's instructions (Fu et al, 2018). Next, the samples were used to enrich poly(A) mRNA, which was fragmented into short pieces by oligo (dT) magnetic beads (Invitrogen).…”