Exosomal miRNA profiling from H5N1 avian influenza virus-infected chickens
Yeojin Hong1, Anh Duc Truong2, Jiae Lee1
1Department of Animal Science and Technology, Chung-Ang University, Anseong, 17546, Republic of Korea.
Abstract:
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.
Insights
This study reveals that exosomal microRNAs (miRNAs) in chickens play a key role in resisting H5N1 avian influenza. Resistant chickens show distinct miRNA expression patterns, offering potential biomarkers for disease resistance.
Area of Science:
- * Avian immunology and virology
- * Molecular biology and genetics
- * Exosome biology and extracellular vesicles
Background:
- * Exosomes are crucial intercellular communicators, carrying diverse biomolecules including microRNAs (miRNAs).
- * Exosomal miRNAs can modulate recipient cell gene expression, influencing host responses.
- * Avian influenza virus (AIV) poses a significant threat to poultry, necessitating research into host resistance mechanisms.
Purpose of the Study:
- * To investigate the role of exosomal miRNAs in the host immune response of Ri chickens to H5N1 highly pathogenic avian influenza virus (HPAIV).
- * To identify differentially expressed miRNAs in resistant versus susceptible chickens upon H5N1 infection.
- * To explore potential exosomal miRNA biomarkers for avian disease resistance.
Main Methods:
- * Genotyping of Mx and BF2 genes to classify Ri chickens into resistant and susceptible groups.
- * H5N1 HPAIV infection model in chickens.
- * Small RNA sequencing of exosomes purified from chicken blood serum.
- * Bioinformatic analysis including miRNA identification, differential expression analysis (edgeR), target gene prediction (miRDB), and gene ontology analysis (gProfiler).
- * Quantitative real-time PCR (qRT-PCR) for miRNA validation.
Main Results:
- * Twenty miRNAs exhibited significantly different expression levels between resistant control and H5N1-infected chickens.
- * Nine miRNAs were upregulated and 11 were downregulated in infected chickens compared to controls.
- * Target gene analysis identified immune-related genes, including cytokines, chemokines, and signaling molecules, particularly within the mitogen-activated protein kinase (MAPK) pathway.
- * qRT-PCR results confirmed sequencing data, showing higher miRNA expression in resistant than susceptible chickens.
Conclusions:
- * Exosomal miRNAs are significantly altered during H5N1 HPAIV infection in chickens.
- * Specific exosomal miRNAs are associated with host resistance to H5N1, potentially by regulating immune-related pathways like MAPK.
- * These findings highlight exosomal miRNAs as potential biomarkers for predicting and enhancing avian disease resistance against HPAIV.
More Related Videos
12:09Avian Influenza Surveillance with FTA Cards: Field Methods, Biosafety, and Transportation Issues Solved
Published on: August 2, 2011
08:10Production of High-Titer Infectious Influenza Pseudotyped Particles with Envelope Glycoproteins from Highly Pathogenic H5N1 and Avian H7N9 Viruses
Published on: January 15, 2020
