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image of Boosting Antiviral Defenses: stat3 Inhibition Enhances Type I Interferon Response to Respiratory Syncytial Virus by Silencing KAP1

Abstract

Introduction

The antiviral effects of type I interferons [IFNs] on respiratory syncytial virus [RSV]-infected airway epithelial cells have been identified. We aim to further reveal the mechanism of stat3 and kruppel-associated box-associated protein 1 [KAP1] in RSV-infected airway epithelial cells.

Methods

Using the A549 cell line, we investigated the impact of RSV infection, KAP1 overexpression, and stat3 inhibition with Stattic. Cell counting kit 8 assay was used to determine the viability, and enzyme-linked immunosorbent assay was applied to measure the levels of IL-6, IL-8, IL-1β, IFN-α, and IFN-β. Viral replication was tested plaque assay. Meanwhile, quantitative real-time reverse transcription polymerase chain reaction or/and western blot were applied to measure the expressions of p-stat3 and KAP1 in the cells.

Results

RSV infection repressed the viability, upregulated p-stat3 and KAP1 expressions, elevated levels of inflammation-related factors [IL-6, IL-8, IL-1β], and type I IFN immune response-associated factors [IFN-α, IFN-β], and promoted viral replication in A549 cells. Stattic attenuated the promoting effect of RSV on inflammation-related factors and viral replication, but enhanced its impact on IFN-α and IFN-β levels in the cells. More importantly, KAP1 overexpression reversed the effects of Stattic on viability, inflammation [IL-6, IL-8, IL-1β], type I IFN immune response [IFN-α, IFN-β], and viral replication in RSV-infected A549 cells.

Conclusion

Our findings unveil the pivotal role of stat3 inhibition in potentiating type I IFN-mediated antiviral responses against RSV in lung epithelial cells, revealing KAP1 as a potential therapeutic target for combating respiratory viral infections.

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2025-05-13
2025-09-14

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References

  1. Rima B. Collins P. Easton A. Fouchier R. Kurath G. Lamb R.A. Lee B. Maisner A. Rota P. Wang L. ICTV virus taxonomy profile: Pneumoviridae. J. Gen. Virol. 2017 98 12 2912 2913 10.1099/jgv.0.000959 29087278
    [Google Scholar]
  2. Coultas J.A. Smyth R. Openshaw P.J. Respiratory syncytial virus (RSV): A scourge from infancy to old age. Thorax 2019 74 10 986 993 10.1136/thoraxjnl‑2018‑212212 31383776
    [Google Scholar]
  3. Domachowske J.B. Anderson E.J. Goldstein M. The future of respiratory syncytial virus disease prevention and treatment. Infect. Dis. Ther. 2021 10 S1 47 60 10.1007/s40121‑020‑00383‑6 33656652
    [Google Scholar]
  4. Villenave R. Broadbent L. Douglas I. Lyons J.D. Coyle P.V. Teng M.N. Tripp R.A. Heaney L.G. Shields M.D. Power U.F. Induction and antagonism of antiviral responses in respiratory syncytial virus-infected pediatric airway epithelium. J. Virol. 2015 89 24 12309 12318 10.1128/JVI.02119‑15 26423940
    [Google Scholar]
  5. Hijano D.R. Vu L.D. Kauvar L.M. Tripp R.A. Polack F.P. Cormier S.A. Role of type I interferon (IFN) in the respiratory syncytial virus (RSV) immune response and disease severity. Front. Immunol. 2019 10 566 10.3389/fimmu.2019.00566 30972063
    [Google Scholar]
  6. Antunes K.H. Fachi J.L. de Paula R. da Silva E.F. Pral L.P. dos Santos A.Á. Dias G.B.M. Vargas J.E. Puga R. Mayer F.Q. Maito F. Zárate-Bladés C.R. Ajami N.J. Sant’Ana M.R. Candreva T. Rodrigues H.G. Schmiele M. Silva Clerici M.T.P. Proença-Modena J.L. Vieira A.T. Mackay C.R. Mansur D. Caballero M.T. Marzec J. Li J. Wang X. Bell D. Polack F.P. Kleeberger S.R. Stein R.T. Vinolo M.A.R. de Souza A.P.D. Microbiota-derived acetate protects against respiratory syncytial virus infection through a GPR43-type 1 interferon response. Nat. Commun. 2019 10 1 3273 10.1038/s41467‑019‑11152‑6 31332169
    [Google Scholar]
  7. Corsello T. Qu Y. Ivanciuc T. Garofalo R.P. Casola A. Antiviral activity of extracellular vesicles derived from respiratory syncytial virus-infected airway epithelial cells. Front. Immunol. 2022 13 886701 10.3389/fimmu.2022.886701 36032066
    [Google Scholar]
  8. Mathieu M. Martin-Jaular L. Lavieu G. Théry C. Specificities of secretion and uptake of exosomes and other extracellular vesicles for cell-to-cell communication. Nat. Cell Biol. 2019 21 1 9 17 10.1038/s41556‑018‑0250‑9 30602770
    [Google Scholar]
  9. Iwai K. Yamamoto S. Yoshida M. Shiba K. Isolation of extracellular vesicles in saliva using density gradient ultracentrifugation. Methods Mol. Biol. 2017 1660 343 350 10.1007/978‑1‑4939‑7253‑1_27 28828669
    [Google Scholar]
  10. Gupta R. Radicioni G. Abdelwahab S. Dang H. Carpenter J. Chua M. Mieczkowski P.A. Sheridan J.T. Randell S.H. Kesimer M. Intercellular communication between airway epithelial cells is mediated by exosome-like vesicles. Am. J. Respir. Cell Mol. Biol. 2019 60 2 209 220 10.1165/rcmb.2018‑0156OC 30230353
    [Google Scholar]
  11. Zhu X. He Z. Yuan J. Wen W. Huang X. Hu Y. Lin C. Pan J. Li R. Deng H. Liao S. Zhou R. Wu J. Li J. Li M. IFITM3-containing exosome as a novel mediator for anti-viral response in dengue virus infection. Cell. Microbiol. 2015 17 1 105 118 10.1111/cmi.12339 25131332
    [Google Scholar]
  12. Chahar H.S. Corsello T. Kudlicki A.S. Komaravelli N. Casola A. Respiratory syncytial virus infection changes cargo composition of exosome released from airway epithelial cells. Sci. Rep. 2018 8 1 387 10.1038/s41598‑017‑18672‑5 29321591
    [Google Scholar]
  13. Stanifer M.L. Guo C. Doldan P. Boulant S. Importance of type I and III interferons at respiratory and intestinal barrier surfaces. Front. Immunol. 2020 11 608645 10.3389/fimmu.2020.608645 33362795
    [Google Scholar]
  14. Randolph K. Hyder U. D’Orso I. KAP1/TRIM28: Transcriptional activator and/or repressor of viral and cellular programs? Front. Cell. Infect. Microbiol. 2022 12 834636 10.3389/fcimb.2022.834636 35281453
    [Google Scholar]
  15. Brattås P.L. Jönsson M.E. Fasching L. Nelander Wahlestedt J. Shahsavani M. Falk R. Falk A. Jern P. Parmar M. Jakobsson J. TRIM28 controls a gene regulatory network based on endogenous retroviruses in human neural progenitor cells. Cell Rep. 2017 18 1 1 11 10.1016/j.celrep.2016.12.010 28052240
    [Google Scholar]
  16. Sheng W. LaFleur M.W. Nguyen T.H. Chen S. Chakravarthy A. Conway J.R. Li Y. Chen H. Yang H. Hsu P.H. Van Allen E.M. Freeman G.J. De Carvalho D.D. He H.H. Sharpe A.H. Shi Y. LSD1 ablation stimulates anti-tumor immunity and enables checkpoint blockade. Cell 2018 174 3 549 563.e19 10.1016/j.cell.2018.05.052 29937226
    [Google Scholar]
  17. Cañadas I. Thummalapalli R. Kim J.W. Kitajima S. Jenkins R.W. Christensen C.L. Campisi M. Kuang Y. Zhang Y. Gjini E. Zhang G. Tian T. Sen D.R. Miao D. Imamura Y. Thai T. Piel B. Terai H. Aref A.R. Hagan T. Koyama S. Watanabe M. Baba H. Adeni A.E. Lydon C.A. Tamayo P. Wei Z. Herlyn M. Barbie T.U. Uppaluri R. Sholl L.M. Sicinska E. Sands J. Rodig S. Wong K.K. Paweletz C.P. Watanabe H. Barbie D.A. Tumor innate immunity primed by specific interferon-stimulated endogenous retroviruses. Nat. Med. 2018 24 8 1143 1150 10.1038/s41591‑018‑0116‑5 30038220
    [Google Scholar]
  18. Li Q. Qin Y. Wang W. Jia M. Zhao W. Zhao C. KAP1-mediated epigenetic suppression in anti-RNA viral responses by direct targeting RIG-I and MDA5. J. Immunol. 2021 207 1 1903 1910 10.4049/jimmunol.2100342 34497149
    [Google Scholar]
  19. Li S. Hu K. Li L. Shen Y. Huang J. Tang L. Zhang L. Shao R. Lu H. Yang Y. Stattic alleviates acute hepatic damage induced by LPS/ d-galactosamine in mice. Innate Immun. 2021 27 2 201 209 10.1177/1753425920988330 33576722
    [Google Scholar]
  20. King C.A. Li X. Barbachano-Guerrero A. Bhaduri-McIntosh S. STAT3 regulates lytic activation of kaposi’s sarcoma-associated herpesvirus. J. Virol. 2015 89 22 11347 11355 10.1128/JVI.02008‑15 26339061
    [Google Scholar]
  21. Lu N. Yang Y. Liu H. Ding X. Ou Y. Xia J. Inhibition of respiratory syncytial virus replication and suppression of RSV-induced airway inflammation in neonatal rats by colchicine. 3 Biotech. 2019 9 11 392 10.1007/s13205‑019‑1917‑z 31656730
    [Google Scholar]
  22. Xu Y. Gao Z. Hu R. Wang Y. Wang Y. Su Z. Zhang X. Yang J. Mei M. Ren Y. Li M. Zhou X. PD-L2 glycosylation promotes immune evasion and predicts anti-EGFR efficacy. J. Immunother. Cancer 2021 9 10 e002699 10.1136/jitc‑2021‑002699 34697216
    [Google Scholar]
  23. Ueba O. Respiratory syncytial virus. I. Concentration and purification of the infectious virus. Acta Med. Okayama 1978 32 4 265 272 153087
    [Google Scholar]
  24. Kim K.S. Kim A.R. Piao Y. Lee J.H. Quan F.S. A rapid, simple, and accurate plaque assay for human respiratory syncytial virus (HRSV). J. Immunol. Meth. 2017 446 15 20 10.1016/j.jim.2017.03.020 28373028
    [Google Scholar]
  25. Pan Y. Wen X. Hao D. Wang Y. Wang L. He G. The role of IL-37 in skin and connective tissue diseases. Biomed. Pharmacoth. 2020 122 109705 10.1016/j.biopha.2019.109705 31918276
    [Google Scholar]
  26. Li Q.Y. Xu H.Y. Yang H.J. Effect of proinflammatory factors TNF-α,IL-1β, IL-6 on neuropathic pain. Zhongguo Zhongyao Zazhi 2017 42 19 3709 3712 29235283
    [Google Scholar]
  27. Hillyer P. Shepard R. Uehling M. Krenz M. Sheikh F. Thayer K.R. Huang L. Yan L. Panda D. Luongo C. Buchholz U.J. Collins P.L. Donnelly R.P. Rabin R.L. Differential responses by human respiratory epithelial cell lines to respiratory syncytial virus reflect distinct patterns of infection control. J. Virol. 2018 92 15 e02202-17 10.1128/JVI.02202‑17 29769339
    [Google Scholar]
  28. Zhang C. Li N. Niu F. Baicalein triazole prevents respiratory tract infection by RSV through suppression of oxidative damage. Microb. Pathog. 2019 131 227 233 10.1016/j.micpath.2019.03.026 30943433
    [Google Scholar]
  29. Feng H. Yi R. Wu S. Wang G. Sun R. Lin L. Zhu S. Nie Z. He Y. Wang S. Wang P. Shu J. Wu L. KAP1 positively modulates influenza A virus replication by interacting with PB2 and NS1 proteins in human lung epithelial cells. Viruses 2022 14 4 689 10.3390/v14040689 35458419
    [Google Scholar]
  30. Kamitani S. Togi S. Ikeda O. Nakasuji M. Sakauchi A. Sekine Y. Krüppel-associated box-associated protein 1 negatively regulates TNF-α-induced NF-κB transcriptional activity by influencing the interactions among STAT3, p300, and NF-κB/p65. J. Immunol. 2011 187 5 2476 2483 10.4049/jimmunol.1003243 21810609
    [Google Scholar]
  31. Xu F. Li X. Yan L. Yuan N. Fang Y. Cao Y. Xu L. Zhang X. Xu L. Ge C. An N. Jiang G. Xie J. Zhang H. Jiang J. Li X. Yao L. Zhang S. Zhou D. Wang J. Autophagy promotes the repair of radiation-induced DNA damage in bone marrow hematopoietic cells via enhanced STAT3 signaling. Radiat. Res. 2017 187 3 382 396 10.1667/RR14640.1 28327001
    [Google Scholar]
  32. Liu S. Liu S. Yu Z. Zhou W. Zheng M. Gu R. Hong J. Yang Z. Chi X. Guo G. Li X. Chen N. Huang S. Wang S. Chen J.L. STAT3 regulates antiviral immunity by suppressing excessive interferon signaling. Cell Rep. 2023 42 7 112806 10.1016/j.celrep.2023.112806 37440406
    [Google Scholar]
  33. Wang K. Liu Z. Zhao M. Zhang F. Wang K. Feng N. Fu F. Li J. Li J. Liu Y. Zhang S. Fan R. Guo H. Pei J. κ-opioid receptor activation promotes mitochondrial fusion and enhances myocardial resistance to ischemia and reperfusion injury via STAT3-OPA1 pathway. Eur. J. Pharmacol. 2020 874 172987 10.1016/j.ejphar.2020.172987 32032598
    [Google Scholar]
  34. Kellar G.G. Barrow K.A. Rich L.M. Debley J.S. Wight T.N. Ziegler S.F. Reeves S.R. Loss of versican and production of hyaluronan in lung epithelial cells are associated with airway inflammation during RSV infection. J. Biol. Chem. 2021 296 100076 10.1074/jbc.RA120.016196 33187989
    [Google Scholar]
  35. Feng Q. Su Z. Song S. Xu H. Zhang B. Yi L. Tian M. Wang H. Histone deacetylase inhibitors suppress RSV infection and alleviate virus-induced airway inflammation. Int. J. Mol. Med. 2016 38 3 812 822 10.3892/ijmm.2016.2691 27460781
    [Google Scholar]
  36. Lazear H.M. Schoggins J.W. Diamond M.S. Shared and distinct functions of type I and type III interferons. Immunity 2019 50 4 907 923 10.1016/j.immuni.2019.03.025 30995506
    [Google Scholar]
  37. Bochnakian A. Zhen A. Zisoulis D.G. Idica A. KewalRamani V.N. Neel N. Daugaard I. Hamdorf M. Kitchen S. Lee K. Pedersen I.M. Interferon-inducible microRNA miR-128 modulates HIV-1 replication by targeting TNPO3 mRNA. J. Virol. 2019 93 20 e00364-19 10.1128/JVI.00364‑19 31341054
    [Google Scholar]
/content/journals/cmm/10.2174/0115665240351066250330235743
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Boosting Antiviral Defenses: stat3 Inhibition Enhances Type I Interferon Response to Respiratory Syncytial Virus by Silencing KAP1
Bentham Science Publishers ; https://doi.org/10.2174/0115665240351066250330235743
/content/journals/cmm/10.2174/0115665240351066250330235743
/content/journals/cmm/10.2174/0115665240351066250330235743
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  • Article Type:     Research Article
Keywords: lung epithelial cells ; Signal transducer and activator of transcription 3 ; kruppel-associated box-associated protein 1 ; immune response ; Stattic ; respiratory syncytial virus ; type I interferon

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