Exosomal miRNA profiling from H5N1 avian influenza virus-infected chickens

Authors: HONG, Y - Chung-Ang University; TROUNG, AD - National Institute Of Veterinary Research; LEE, J - Chung-Ang University; VU, TH - Chung-Ang University; LEE, S - Chung-Ang University; SONG, KD - Jeonbuk National University; Lillehoj, Hyun; HONG, YH - Chung-Ang University

Submitted to: Scientific Reports
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/2/2021
Publication Date: 3/3/2021
Publication URL: https://handle.nal.usda.gov/10113/7299277
Citation: Hong, Y., Troung, A., Lee, J., Vu, T., Lee, S., Song, K., Lillehoj, H.S., Hong, Y. 2021. Exosomal miRNA profiling from H5N1 avian influenza virus-infected chickens. Scientific Reports. https://doi.org/10.1186/s13567-021-00892-3.
DOI: https://doi.org/10.1186/s13567-021-00892-3

Interpretive Summary: Information concerning poultry immune system and its molecular interactions with pathogens that modulate physiological responses in disease conditions in chickens is limited. ARS scientists collaborated with Korean scientists to investigate the mechanisms of viral pathogenesis using avian influenza virus as a model in order to develop a logical control strategy against avian influenza infections. In this trial which was carried out in South Korea, microRNA (miRNA) from chickens infected with avian influenza were sequenced and analysed. Twenty miRNAs showed significantly different expression patterns that were associated with resistant phenotype and the following gene analysis showed that many miRNA changes were related to various immune-related genes, such as cytokines, chemokines, and signalling molecules. Detailed investigation of these genes will lead to better understanding of how poultry immune system interact with avian influenza. Taken together, this study provides an insight into the different molecular signalling mechanisms that virus-resistant chickens use in defence against avian influenza virus. Furthermore, the finding of this study improves our understanding of anti-viral host immune response that will eventually facilitate the development of biomarkers for avian influenza and an avian influenza vaccine.

Technical 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 infected with H5N1, a highly pathogenic avian influenza virus (HPAIV). Exosomes were purified 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. This study will help to better understand the host immune response, particularly exosomal miRNA expression against HPAIV H5N1. Finally, these results could help develop an avian influenza vaccine and determine biomarkers for disease resistance.