Cyclophilin inhibitors restrict Middle East respiratory syndrome coronavirus via interferon-lambda in vitro and in mice

While severe coronavirus infections, including Middle East respiratory syndrome coronavirus (MERS-CoV), cause lung injury with high mortality rates, protective treatment strategies are not approved for clinical use.We elucidated the molecular mechanisms by which the cyclophilin inhibitors cyclosporin A (CsA) and alisporivir (ALV) restrict MERS-CoV to validate their suitability as readily available therapy in MERS-CoV infection.Calu-3 cells and primary human alveolar epithelial cells (hAECs) were infected with MERS-CoV and treated with CsA or ALV or inhibitors targeting cyclophilin inhibitor-regulated molecules including calcineurin, nuclear factor of activated T-cells (NFATs) or mitogen-activated protein kinases. Novel CsA-induced pathways were identified by RNA sequencing and manipulated by gene knockdown or neutralising antibodies. Viral replication was quantified by quantitative real-time PCR and 50% tissue culture infective dose. Data were validated in a murine MERS-CoV infection model.Both CsA and ALV reduced MERS-CoV titres and viral RNA replication in Calu-3 cells and hAECs, improving epithelial integrity. While neither calcineurin nor NFAT inhibition reduced MERS-CoV propagation, blockade of c-Jun N-terminal kinase diminished infectious viral particle release but not RNA accumulation. Importantly, CsA induced interferon regulatory factor 1 (IRF1), a pronounced type III interferon (IFNlambda) response and expression of antiviral genes. Downregulation of IRF1 or IFNlambda increased MERS-CoV propagation in the presence of CsA. Importantly, oral application of CsA reduced MERS-CoV replication in vivo, correlating with elevated lung IFNlambda levels and improved outcome.We provide evidence that cyclophilin inhibitors efficiently decrease MERS-CoV replication in vitro and in vivo via upregulation of inflammatory antiviral cell responses, in particular IFNlambda. CsA might therefore represent a promising candidate for treating MERS-CoV infection.

  • Sauerhering, L.
  • Kupke, A.
  • Meier, L.
  • Dietzel, E.
  • Hoppe, J.
  • Gruber, A. D.
  • Gattenloehner, S.
  • Witte, B.
  • Fink, L.
  • Hofmann, N.
  • Zimmermann, T.
  • Goesmann, A.
  • Nist, A.
  • Stiewe, T.
  • Becker, S.
  • Herold, S.
  • Peteranderl, C.

Keywords

  • Alveolar Epithelial Cells/drug effects/metabolism/virology
  • Animals
  • Calcineurin Inhibitors/pharmacology
  • Cell Culture Techniques
  • Coronavirus Infections/metabolism/*prevention & control
  • Cyclophilins/*antagonists & inhibitors
  • Cyclosporine/*pharmacology
  • Disease Models, Animal
  • Humans
  • Interferon Regulatory Factor-1/drug effects/metabolism
  • Interferons/drug effects/*metabolism
  • Mice
  • Middle East Respiratory Syndrome Coronavirus/*drug effects/physiology
  • Virus Replication/drug effects
Publication details
DOI: 10.1183/13993003.01826-2019
Journal: Eur Respir J
Number: 5
Work Type: Original
Access number: 32616594
See publication on PubMed
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