Journal
VETERINARY RESEARCH
Volume 52, Issue 1, Pages -Publisher
BMC
DOI: 10.1186/s13567-021-00892-3
Keywords
HPAIV; Exosomes; Small RNA sequencing; miRNA
Categories
Funding
- Vietnam National Foundation for Science and Technology Development (NAFOSTED) [106.022019.01]
- National Research Foundation of the Republic of Korea [NRF-2021R1A2C2005236]
- NIFA from the USDA [2017-6701526793]
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This study identified significantly differentially expressed miRNAs in resistant chickens compared to susceptible chickens after H5N1 infection, with a focus on regulation of immune-related genes, particularly the MAPK pathway molecules. The results showed higher miRNA expression levels in resistant chickens than susceptible chickens, indicating a potential for using exosomal miRNA expression as biomarkers for disease resistance against HPAIV H5N1.
Exosomes are membrane vesicles containing proteins, lipids, DNA, mRNA, and micro RNA (miRNA). Exosomal miRNA from donor cells can regulate the gene expression of recipient cells. Here, Ri chickens were divided into resistant (Mx/A; BF2/B21) and susceptible (Mx/G; BF2/B13) trait by genotyping of Mx and BF2 genes. Then, Ri chickens were infected with H5N1, a highly pathogenic avian influenza virus (HPAIV). Exosomes were purified from blood serum of resistant chickens for small RNA sequencing. Sequencing data were analysed using FastQCv0.11.7, Cutadapt 1.16, miRBase v21, non-coding RNA database, RNAcentral 10.0, and miRDeep2. Differentially expressed miRNAs were determined using statistical methods, including fold-change, exactTest using edgeR, and hierarchical clustering. Target genes were predicted using miRDB. Gene ontology analysis was performed using gProfiler. Twenty miRNAs showed significantly different expression patterns between resistant control and infected chickens. Nine miRNAs were up-regulated and 11 miRNAs were down-regulated in the infected chickens compared with that in the control chickens. In target gene analysis, various immune-related genes, such as cytokines, chemokines, and signalling molecules, were detected. In particular, mitogen-activated protein kinase (MAPK) pathway molecules were highly controlled by differentially expressed miRNAs. The result of qRT-PCR for miRNAs was identical with sequencing data and miRNA expression level was higher in resistant than susceptible chickens. This study will help to better understand the host immune response, particularly exosomal miRNA expression against HPAIV H5N1 and could help to determine biomarkers for disease resistance.
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