본문 바로가기
KERD
논문KCI 등재

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)

  1. Lu M, "Exosome-based small RNA delivery : progress and prospects", Asian J Pharm, 13, pp.1~11 (2018)[DOI]
  2. Bakhshandeh B, "A comprehensive review on exosomes and microvesicles as epigenetic factors", Curr Stem Cell Res Ther, 12, pp.31~36 (2017)[DOI]
  3. Mathivanan S, "Exosomes : extracellular organelles important in intercellular communication", J Proteomics, 73, pp.1907~1920 (2010)[DOI]
  4. Gross JC, "Active Wnt proteins are secreted on exosomes", Nat Cell Biol, 14, pp.1036~1045 (2012)[DOI]
  5. Tkach M, "Communication by extracellular vesicles : where we are and where we need to go", Cell, 164, pp.1226~1232 (2016)[DOI]
  6. Bartel DP, "MicroRNAs : genomics, biogenesis, mechanism, and function", Cell, 116, pp.281~297 (2004)[DOI]
  7. He L, "MicroRNAs : small RNAs with a big role in gene regulation", Nat Rev Genet, 5, pp.522~531 (2004)[DOI]
  8. Melo SA, "Cancer exosomes perform cell-independent microRNA biogenesis and promote tumorigenesis", Cancer Cell, 26, pp.707~721 (2014)[DOI]
  9. Hannafon BN, "Intercellular communication by exosome-derived microRNAs in cancer", Int J Mol Sci, 14, pp.14240~14269 (2013)[DOI]
  10. 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]
  11. 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]
  12. 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]
  13. 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]
  14. 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]
  15. 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]
  16. Paranjpe M, "Nanoparticle-mediated pulmonary drug delivery : a review", Int J Mol Sci, 15, pp.5852~5873 (2014)[DOI]
  17. Tan S, "Cell or cell membrane-based drug delivery systems", Theranostics, 5, pp.863~881 (2015)[DOI]
  18. Wu Y, "MiR-20a-5p is multifunctional regulator in chickens immune responses induced by NDV, IBDV and AIV vaccines respectively", Res Sq (2021)[DOI]
  19. 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]
  20. 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]
  21. 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]
  22. Hong Y, "Exosomal miRNA profiling from H5N1 avian influenza virus-infected chickens", Vet Res, 52, pp.36~ (2021)[DOI]
  23. 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]
  24. Hong Y, "Immunomodulatory effects of avian β-defensin 5 in chicken macrophage cell line", Res Vet Sci, 132, pp.81~87 (2020)[DOI]
  25. 김하늘, "Exosome-mediated Let7c-5p Delivery for Breast Cancer Therapeutic Development", Biotechnology and Bioprocess Engineering, 25(4), pp.513~520 (2020), 한국생물공학회[DOI]
  26. 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]
  27. Fu X, "miR-20a-5p", TGFBR2axis affects pro-inflammatory macrophages and aggravates liver fibrosis, 2020, pp.10, 107~
  28. Xiong Y, "MiR-20a is upregulated in anaplastic thyroid cancer and targets LIMK1", PLoS One, 9, pp.e96103~ (2014)[DOI]
  29. 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
  30. 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]
  31. Laengsri V, "Cervical cancer markers : epigenetics and microRNAs", Lab Med, 49, pp.97~111 (2018)[DOI]
  32. Hirayama D, "The phagocytic function of macrophage-enforcing innate immunity and tissue homeostasis", Int J Mol Sci, 19, pp.92~ (2018)[DOI]
  33. Alexopoulou L, "Recognition of double-stranded RNA and activation of NF-κB by toll-like receptor 3", Nature, 413, pp.732~738 (2001)[DOI]
  34. Kawai T, "Toll-like receptor and RIG-I-like receptor signaling", Ann NY Acad Sci, 1143, pp.1~20 (2008)[DOI]
  35. Jorgovanovic D, "Roles of IFN-γin tumor progression and regression : a review", Biomark Res, 8, pp.49~ (2020)[DOI]
  36. Tang M, "Interferon-gamma-mediated osteoimmunology", Front Immunol, 9, pp.1508~ (2018)[DOI]
  37. 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]
  38. Johnsen IB, "Toll-like receptor 3-elicited MAPK activation induces stabilization of interferon-β mRNA", Cytokine, 57, pp.337~346 (2012)[DOI]
  39. Peroval MY, "A critical role for MAPK signalling pathways in the transcriptional regulation of toll like receptors", PloS One, 8, pp.e51243~ (2013)[DOI]
  40. 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]
  41. Richer MJ, "Toll-like receptor 3 signaling on macrophages is required for survival following coxsackievirus B4 infection", PloS One, 4, pp.e4127~ (2009)[DOI]
  42. Idriss HT, "TNFα and the TNF receptor superfamily : Structure-function relationship(s)", Microsc Res Tech, 50, pp.184~195 (2000)[DOI]
  43. Liu Q, "The cytokine storm of severe influenza and development of immunomodulatory therapy", Cell Mol Immunol, 13, pp.3~10 (2016)[DOI]
  44. Tisoncik JR, "Into the eye of the cytokine storm", Microbiol Mol Biol Rev, 76, pp.16~32 (2012)[DOI]
  45. D'Elia RV, "Targeting the"cytokine storm"for therapeutic benefit", Clin Vaccine Immunol, 20, pp.319~327 (2013)[DOI]
  46. Gu Y, "The mechanism behind influenza virus cytokine storm", Viruses, 13, pp.1362~ (2021)[DOI]
  47. Yu J, "Regulation of host immune responses against influenza a virus infection by mitogenactivated protein kinases(MAPKs)", Microorganisms, 8, pp.1067~ (2020)[DOI]

관련 논문AI추천

MicroRNA expression profiling in the lungs of genetically different Ri chicken lines against the highly pathogenic avian influenza H5N1 virus이수연, 허주비, 홍여진, 홍영호, Anh Duc Truong, Hyun S. Lillehoj, Suyeon Kang, Thi Hao Vu한국축산학회 · 한국축산학회지2023 · 제65권 제4호Distribution and differential expression of microRNAs in the intestinal mucosal layer of necrotic enteritis induced Fayoumi chickens홍영호, 데이벤드란렝가라즈, Anh Duc Truong, Hyun S. Lillehoj, Jihye Ban아세아·태평양축산학회 · Animal Bioscience2017 · 제30권 제7호The highly pathogenic H5N1 avian influenza virus induces the mitogen-activated protein kinase signaling pathway in the trachea of two Ri chicken lines부 티 하오, 홍영호, Heo Jubi, Hong Yeojin, Lee Sooyeon, Lillehoj,Hyun S., Truong Anh Duc아세아·태평양축산학회 · Animal Bioscience2022 · 제35권 제7호Enrichment and verification of differentially expressed miRNAs in bursa of Fabricius in two breeds of duckChunchun Han, He-he Liu, Hua He, Jiwen Wang, Jun Luo, Junying Liu, Tao Zhang아세아·태평양축산학회 · Animal Bioscience2017 · 제30권 제7호Expression patterns of innate immunity-related genes in response to polyinosinic:polycytidylic acid (poly[I:C]) stimulation in DF-1 chicken fibroblast cells장현준, Ki-Duk Song한국축산학회 · 한국축산학회지2020 · 제62권 제3호In vitro analysis of antiviral immune response against avian influenza virus in chicken tracheal epithelial cells부 티 하오, 홍영호, 츠엉앵득, Heo Jubi, Kim CH아세아·태평양축산학회 · Animal Bioscience2024 · 제37권 제12호

이 간행물의 다른 논문 (15건)