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https://doi.org/10.5713/ab.22.0373    [Accepted] Published online January 11, 2023.
Exosome-mediated delivery of gga-miR-20a-5p regulates immune response of chicken macrophages by targeting IFNGR2, MAPK1, MAP3K5, and MAP3K14
Yeojin Hong1  , Jubi Heo1  , Suyeon Kang1  , Thi Hao Vu1  , Hyun S Lillehoj2  , Yeong Ho Hong1,* 
1Department of Animal Science and Technology, Chung-Ang University, Anseong 17546, Korea
2Animal Biosciences and Biotechnology Laboratory, Agricultural Research Services, United States Department of Agriculture, Beltsville, MD 20705, USA
Correspondence:  Yeong Ho Hong, Tel: +82-31-670-3025, Fax: +82-31-675-3108, Email: yhong@cau.ac.kr
Received: 20 September 2022   • Revised: 21 November 2022   • Accepted: 24 December 2022
Abstract
Objective
This study aims to evaluate the target genes of gga-miR-20a-5p and the regulated immune responses in the chicken macrophage cell line, HD11, by the exosome-mediated delivery of miR-20a-5p.
Methods
Exosomes were purified from the chicken macrophage cell line HD11. Then, mimic gga-miR-20p or negative control miRNA were internalized into HD11 exosomes. HD11 cells were transfected with gga-miR-20a-5p or negative control miRNA containing exosomes. After 44 h of transfection, cells were incubated with or without 5 μg/mL poly(I:C) for 4 h. Then, expression of target genes and cytokines was evaluated by qRT-PCR.
Results
Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted IFNGR2, MAPK1, MAP3K5, and MAP3K14. Moreover, the exosome-mediated delivery of gga-miR-20a-5p successfully repressed the expression of IFNGR2, MAPK1, MAP3K5, and MAP3K14 in HD11 cells. The expressions of interferon-stimulated genes (ISGs) (MX1, EIF2A, and OASL) and proinflammatory cytokines (IFNG, IL1B, and TNFA) were also downregulated by exosomal miR-20a-5p. In addition, the proliferation of HD11 cells was increased by exosomal miR-20a-5p.
Conclusion
The exosome-mediated delivery of gga-miR-20a-5p regulated immune responses by controlling the MAPK and apoptotic signaling pathways. Furthermore, we expected that exosomal miR-20a-5p could maintain immune homeostasis against HPAIV H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.
Keywords: Apoptosis; Exosomes; miRNAs; miR-20a-5p; MAPK Signaling Pathway
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