Exosome-mediated delivery of gga-miR-20a-5p regulates immune response of chicken macrophages by targeting IFNGR2, MAPK1, MAP3K5, and MAP3K14
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
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 quantitative realtime polymerase chain reaction. Results: Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted interferon gamma receptor 2 (IFNGR2), mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase kinase kinase 5 (MAP3K5), and mitogenactivated protein kinase kinase kinase 14 (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 (MX dynamin like GTPase 1 [MX1], eukaryotic translation initiation factor 2A [EIF2A], and oligoadenylate synthase-like [OASL]) and proinflammatory cytokines (interferongamma [IFNG], interleukin-1 beta [IL1B], and tumor necrosis factor-alpha [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 highly pathogenic avian influenza virus H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.
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 quantitative realtime polymerase chain reaction. Results: Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted interferon gamma receptor 2 (IFNGR2), mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase kinase kinase 5 (MAP3K5), and mitogenactivated protein kinase kinase kinase 14 (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 (MX dynamin like GTPase 1 [MX1], eukaryotic translation initiation factor 2A [EIF2A], and oligoadenylate synthase-like [OASL]) and proinflammatory cytokines (interferongamma [IFNG], interleukin-1 beta [IL1B], and tumor necrosis factor-alpha [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 highly pathogenic avian influenza virus H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.
참고문헌 (47)
- Lu M, "Exosome-based small RNA delivery : progress and prospects", Asian J Pharm, 13, pp.1~11 (2018)[DOI]
- Bakhshandeh B, "A comprehensive review on exosomes and microvesicles as epigenetic factors", Curr Stem Cell Res Ther, 12, pp.31~36 (2017)[DOI]
- Mathivanan S, "Exosomes : extracellular organelles important in intercellular communication", J Proteomics, 73, pp.1907~1920 (2010)[DOI]
- Gross JC, "Active Wnt proteins are secreted on exosomes", Nat Cell Biol, 14, pp.1036~1045 (2012)[DOI]
- Tkach M, "Communication by extracellular vesicles : where we are and where we need to go", Cell, 164, pp.1226~1232 (2016)[DOI]
- Bartel DP, "MicroRNAs : genomics, biogenesis, mechanism, and function", Cell, 116, pp.281~297 (2004)[DOI]
- He L, "MicroRNAs : small RNAs with a big role in gene regulation", Nat Rev Genet, 5, pp.522~531 (2004)[DOI]
- Melo SA, "Cancer exosomes perform cell-independent microRNA biogenesis and promote tumorigenesis", Cancer Cell, 26, pp.707~721 (2014)[DOI]
- Hannafon BN, "Intercellular communication by exosome-derived microRNAs in cancer", Int J Mol Sci, 14, pp.14240~14269 (2013)[DOI]
- Li XJ, "Exosomal MicroRNA MiR-1246 promotes cell proliferation, invasion and drug resistance by targeting CCNG2 in breast cancer", Cell Physiol Biochem, 44, pp.1741~1748 (2017)[DOI]
- Jeong K, "Exosome-mediated microrna-497 delivery for anti-cancer therapy in a microfluidic 3d lung cancer model", Lab Chip, 20, pp.548~557 (2020)[DOI]
- Lee GW, "Exosome mediated transfer of miRNA-140 promotes enhanced chondrogenic differentiation of bone marrow stem cells for enhanced cartilage repair and regeneration", J Cell Biochem, 121, pp.3642~3652 (2020)[DOI]
- Rezaei R, "Exosome-mediated delivery of functionally active miRNA-375-3p mimic regulate epithelial mesenchymal transition(EMT)of colon cancer cells", Life Sci, 269, pp.119035~ (2021)[DOI]
- Johnsen KB, "A comprehensive overview of exosomes as drug delivery vehicles—endogenous nanocarriers for targeted cancer therapy", Biochim Biophys Acta Rev Cancer, 1846, pp.75~87 (2014)[DOI]
- Lamichhane TN, "Exogenous DNA loading into extracellular vesicles via electroporation is size-dependent and enables limited gene delivery", Mol Pharmaceutics, 12, pp.3650~3657 (2015)[DOI]
- Paranjpe M, "Nanoparticle-mediated pulmonary drug delivery : a review", Int J Mol Sci, 15, pp.5852~5873 (2014)[DOI]
- Tan S, "Cell or cell membrane-based drug delivery systems", Theranostics, 5, pp.863~881 (2015)[DOI]
- Wu Y, "MiR-20a-5p is multifunctional regulator in chickens immune responses induced by NDV, IBDV and AIV vaccines respectively", Res Sq (2021)[DOI]
- Philippe L, "MiR-20a regulates ASK1 expression and TLR4-dependent cytokine release in rheumatoid fibroblast-like synoviocytes", Ann Rheum Dis, 72, pp.1071~1079 (2013)[DOI]
- Zhang G, "Down-regulation of miR-20a-5p triggers cell apoptosis to facilitate mycobacterial clearance through targeting JNK2 in human macrophages", Cell Cycle, 15, pp.2527~2538 (2016)[DOI]
- Chang R, "MicroRNA-20a-5p suppresses IL-17 production by targeting OSM and CCL1 in patients with Vogt-Koyanagi-Harada disease", Br J Ophthalmol, 102, pp.282~290 (2018)[DOI]
- Hong Y, "Exosomal miRNA profiling from H5N1 avian influenza virus-infected chickens", Vet Res, 52, pp.36~ (2021)[DOI]
- Klasing KC, "Influence of cell sources, stimulating agents, and incubation conditions on release of interleukin-1from chicken macrophages", Dev Comp Immunol, 11, pp.385~394 (1987)[DOI]
- Hong Y, "Immunomodulatory effects of avian β-defensin 5 in chicken macrophage cell line", Res Vet Sci, 132, pp.81~87 (2020)[DOI]
- 김하늘, "Exosome-mediated Let7c-5p Delivery for Breast Cancer Therapeutic Development", Biotechnology and Bioprocess Engineering, 25(4), pp.513~520 (2020), 한국생물공학회[DOI]
- Livak KJ, "Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method", Methods, 25, pp.402~408 (2001)[DOI]
- Fu X, "miR-20a-5p", TGFBR2axis affects pro-inflammatory macrophages and aggravates liver fibrosis, 2020, pp.10, 107~
- Xiong Y, "MiR-20a is upregulated in anaplastic thyroid cancer and targets LIMK1", PLoS One, 9, pp.e96103~ (2014)[DOI]
- Wang M, "miR-17-5p", 20a are important markers for gastric cancer and murine double minute 2 participates in their functional regulation, 2013, pp.49, 2010~2021
- Guo L, "Breast cancer cell-derived exosomal miR-20a-5p promotes the proliferation and differentiation of osteoclasts by targeting SRCIN1", Cancer Med, 8, pp.5687~5701 (2019)[DOI]
- Laengsri V, "Cervical cancer markers : epigenetics and microRNAs", Lab Med, 49, pp.97~111 (2018)[DOI]
- Hirayama D, "The phagocytic function of macrophage-enforcing innate immunity and tissue homeostasis", Int J Mol Sci, 19, pp.92~ (2018)[DOI]
- Alexopoulou L, "Recognition of double-stranded RNA and activation of NF-κB by toll-like receptor 3", Nature, 413, pp.732~738 (2001)[DOI]
- Kawai T, "Toll-like receptor and RIG-I-like receptor signaling", Ann NY Acad Sci, 1143, pp.1~20 (2008)[DOI]
- Jorgovanovic D, "Roles of IFN-γin tumor progression and regression : a review", Biomark Res, 8, pp.49~ (2020)[DOI]
- Tang M, "Interferon-gamma-mediated osteoimmunology", Front Immunol, 9, pp.1508~ (2018)[DOI]
- Kim You-Me, "Type I and III interferon responses in SARS-CoV-2 infection", Experimental and Molecular Medicine, 53, pp.1~11 (2021), 생화학분자생물학회[DOI]
- Johnsen IB, "Toll-like receptor 3-elicited MAPK activation induces stabilization of interferon-β mRNA", Cytokine, 57, pp.337~346 (2012)[DOI]
- Peroval MY, "A critical role for MAPK signalling pathways in the transcriptional regulation of toll like receptors", PloS One, 8, pp.e51243~ (2013)[DOI]
- Sun R, "Toll-like receptor 3(TLR3)induces apoptosis via death receptors and mitochondria by up-regulating the transactivating p63 isoform α(TAP63α)", J Biol Chem, 286, pp.15918~15928 (2011)[DOI]
- Richer MJ, "Toll-like receptor 3 signaling on macrophages is required for survival following coxsackievirus B4 infection", PloS One, 4, pp.e4127~ (2009)[DOI]
- Idriss HT, "TNFα and the TNF receptor superfamily : Structure-function relationship(s)", Microsc Res Tech, 50, pp.184~195 (2000)[DOI]
- Liu Q, "The cytokine storm of severe influenza and development of immunomodulatory therapy", Cell Mol Immunol, 13, pp.3~10 (2016)[DOI]
- Tisoncik JR, "Into the eye of the cytokine storm", Microbiol Mol Biol Rev, 76, pp.16~32 (2012)[DOI]
- D'Elia RV, "Targeting the"cytokine storm"for therapeutic benefit", Clin Vaccine Immunol, 20, pp.319~327 (2013)[DOI]
- Gu Y, "The mechanism behind influenza virus cytokine storm", Viruses, 13, pp.1362~ (2021)[DOI]
- Yu J, "Regulation of host immune responses against influenza a virus infection by mitogenactivated protein kinases(MAPKs)", Microorganisms, 8, pp.1067~ (2020)[DOI]
