Cynarin blocks Ebola virus replication by counteracting VP35 inhibition of interferon-beta production

Angela Corona, Elisa Fanunza, Cristiano Salata, Melody Jane Morwitzer, Simona Distinto, Luca Zinzula, Cinzia Sanna, Aldo Frau, Gian Luca Daino, Marina Quartu, Orazio Taglialatela-Scafati, Daniela Rigano, St Patrick Reid, Alì Mirazimi, Enzo Tramontano

Research output: Contribution to journalArticlepeer-review

Abstract

Ebola virus (EBOV) is one of the deadliest infective agents whose lethality is linked to the ability to efficiently bypass the host's innate antiviral response. EBOV multifunctional protein VP35 plays a major role in viral replication both as polymerase cofactor and interferon (IFN) antagonist. By hiding the non-self 5′-ppp dsRNA from the cellular receptor RIG-I, VP35 prevents its activation and inhibits IFN-β production. Blocking VP35-dsRNA interaction and IFN-β suppression is a validated drug target. We screened a library of natural extracts and found that cynarin inhibits dsRNA-VP35 binding with an IC50 value of 8.5 μM. It reverts the EBOV VP35 inhibition of IFN-β production, while it does not induce IFN production by itself. Docking experiments suggest that the molecule can bind on the end-capping pocket of VP35 C-terminal Interferon Inhibitory domain (IID), and differential scanning fluorimetry confirmed that cynarin interacts with VP35-IID with a KD of 12 μM. Cynarin was further tested in an EBOV minigenome assay but did not inhibit VP35 polymerase cofactor activity. When evaluated during challenge of IFN-susceptible A549 cells with EBOV isolate derived from the 2014 West African outbreak, cynarin was able to inhibit viral replication with an EC50 value of 9.1 μM, showing no significant cytotoxicity. Our findings show that cynarin blocks EBOV replication by acting directly on VP35 and subverting its IFN antagonism function but not cofactor function, and as such identify the first EBOV inhibitor with this mode of action.

Original languageEnglish (US)
Article number105251
JournalAntiviral Research
Volume198
DOIs
StatePublished - Feb 2022

Keywords

  • Cynarin
  • Dicaffeolyliquinic acid
  • Ebola virus VP35
  • Ebola virus inhibition
  • RNA binding protein inhibitor
  • Type I IFN antagonist
  • Viral immune antagonist

ASJC Scopus subject areas

  • Pharmacology
  • Virology

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