GNS561 exhibits potent in vitro antiviral activity against SARS-CoV-2 through autophagy inhibition

  1. Philippe Halfon
  2. Eloïne Bestion
  3. Keivan Zandi
  4. Julien Andreani
  5. Jean-Pierre Baudoin
  6. Bernard La Scola
  7. Jean-Louis Mege
  8. Soraya Mezouar
  9. Raymond F. Schinazi

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Abstract

Since December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2/2019-nCoV) has spread quickly worldwide, with more than 29 million cases and 920,000 deaths. Interestingly, coronaviruses were found to subvert and hijack the autophagic process to allow their viral replication. One of the spotlights had been focused on the autophagy inhibitors as a target mechanism effective in the inhibition of SARS-CoV-2 infection. Consequently, chloroquine (CQ) and hydroxychloroquine (HCQ), a derivative of CQ, was suggested as the first potentially be therapeutic strategies as they are known to be autophagy inhibitors. Then, they were used as therapeutics in SARS-CoV-2 infection along with remdesivir, for which the FDA approved emergency use authorization. Here, we investigated the antiviral activity and associated mechanism of GNS561, a small basic lipophilic molecule inhibitor of late-stage autophagy, against SARS-CoV-2. Our data indicated that GNS561 showed the highest antiviral effect for two SARS-CoV-2 strains compared to CQ and remdesivir. Focusing on the autophagy mechanism, we showed that GNS561, located in LAMP2-positive lysosomes, together with SARS-CoV-2, blocked autophagy by increasing the size of LC3-II spots and the accumulation of autophagic vacuoles in the cytoplasm with the presence of multilamellar bodies characteristic of a complexed autophagy. Finally, our study revealed that the combination of GNS561 and remdesivir was associated with a strong synergistic antiviral effect against SARS-CoV-2. Overall, our study highlights GNS561 as a powerful drug in SARS-CoV-2 infection and supports that the hypothesis that autophagy inhibitors could be an alternative strategy for SARS-CoV-2 infection.

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  1. ScreenIT

    SciScore for 10.1101/2020.10.06.327635: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    RandomizationSix randomly selected microscopic fields of cells were acquired and collected by Zen 3.0 (Blue Edition) software (Zeiss).
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Then, the coverslips were incubated in blocking buffer for 45 min at room temperature with antibodies against LC3-II (Sigma Aldrich, 1:200)
    LC3-II
    suggested: None
    The secondary antibodies used to reveal LC3 and LAMP2 were Alexa 594-labeled anti-rabbit secondary antibody and Alexa 546-labeled anti-mouse secondary antibody (Invitrogen, USA), respectively.
    LC3
    suggested: None
    LAMP2
    suggested: None
    anti-rabbit
    suggested: None
    anti-mouse
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Vero E6 cells cultured on coverslips were initially treated or not by a fluorescent analog of GNS561, GNS561G, for 2 hours and were then infected by SARS-CoV-2 (IHU-MI6, MOI 0.1).
    Vero E6
    suggested: None
    Software and Algorithms
    SentencesResources
    Six randomly selected microscopic fields of cells were acquired and collected by Zen 3.0 (Blue Edition) software (Zeiss).
    Zen
    suggested: None

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: We found the following clinical trial numbers in your paper:

    IdentifierStatusTitle
    NCT03316222RecruitingStudy of GNS561 in Patients With Liver Cancer
    NCT04333914RecruitingProspective Study in Patients With Advanced or Metastatic Ca…

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

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  2. Version published to 10.1101/2020.10.06.327635v1 on bioRxiv