A Small interfering RNA lead targeting RNA-dependent RNA-polymerase effectively inhibit the SARS-CoV-2 infection in Golden Syrian hamster and Rhesus macaque

  1. Se Hun Gu
  2. Chi Ho Yu
  3. Youngjo Song
  4. Na Young Kim
  5. Euni Sim
  6. Jun Young Choi
  7. Dong Hyun Song
  8. Gyeung Haeng Hur
  9. Young Kee Shin
  10. Seong Tae Jeong

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Abstract

A small interfering RNA (siRNA) inhibitors have demonstrated the novel modality for suppressing infectious diseases. Sixty-one siRNA molecules, predicted by the bioinformatics programs, were screened for the possibility of treating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using an in vitro plaque assay. Among six siRNA leads with the efficacy of reducing plaque number, the siRNA targeting RNA-dependent RNA polymerase (RdRp) showed a reduction in SARS-CoV-2 infection-induced fever and virus titer in the Golden Syrian hamster and rhesus macaque. These results suggest the potential for RdRp targeting siRNA as a new treatment for the coronavirus disease 2019 (COVID-19).

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  1. ScreenIT

    SciScore for 10.1101/2020.07.07.190967: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: All inoculations and handling of the animals, as well as the method of euthanasia and collection of tissues, were performed according to well-established protocols approved by the Institutional Animal Care and Use Committee of the Agency for Defense Development (ADD-IACUC-20-12 and ADD-IACUC-20-13).
    RandomizationFirst, we confirmed whether Syrian hamsters were infected with SARS-CoV-2; 6-week-old male Syrian hamsters were randomly segregated into 5 groups.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableFirst, we confirmed whether Syrian hamsters were infected with SARS-CoV-2; 6-week-old male Syrian hamsters were randomly segregated into 5 groups.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    In vitro efficacy test using Vero E6 cells (siRNA transfection): Vero E6 cells were inoculated into a 24-well plate, 1 mL each, through a 10% FBS DMEM without penicillin/streptomycin to ensure that the 24-well plate is 80–90% confluent.
    Vero E6
    suggested: None
    Software and Algorithms
    SentencesResources
    Potential siRNA sequences targeting viral genes were predicted using the programs siDirect (http://sidirect2.rnai.jp/) and VIRsiRNApred (http://crdd.osdd.net/servers/virsirnapred/).
    siDirect
    suggested: (siDirect, RRID:SCR_004853)

    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: No clinical trial numbers were referenced.

    Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    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

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. ScreenIT

    SciScore for 10.1101/2020.07.07.190967: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementAll inoculations and handling of the animals , as well as the method of euthanasia and collection of tissues , were performed according to well-established protocols approved by the Institutional Animal Care and Use Committee of the Agency for Defense Development ( ADD-IACUC-20-12 and ADDIACUC-20-13) .RandomizationFirst , we confirmed whether Syrian hamsters were infected with SARS-CoV-2; 6-week-old male Syrian hamsters were randomly segregated into 5 groups .Blindingnot detected.Power Analysisnot detected.Sex as a biological variableFirst , we confirmed whether Syrian hamsters were infected with SARS-CoV-2; 6-week-old male Syrian hamsters were randomly segregated into 5 groups .Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The Syrian hamster model was used for demonstrating protective efficacy of neutralizing antibodies against SARS-CoV-2.
    SARS-CoV-2
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The next day , the Vero E6 cells were transfected with 100 nM siRNA using Lipofectamine RNAiMAX transfection reagent ( Invitrogen , USA) , according to the manufacturer’s instructions .
    Vero E6
    suggested: None
    Wang et al presented that SARS-CoV replication was efficiently inhibited by siRNA in Vero cells.
    Vero
    suggested: CLS Cat# 605372/p622_VERO, CVCL_0059
    Software and Algorithms
    SentencesResources
    Potential siRNA sequences targeting viral genes were predicted using the programs siDirect ( http://sidirect2.rnai.jp/ ) and VIRsiRNApred ( http://crdd.osdd.net/servers/virsirnapred/).
    siDirect
    suggested: (siDirect, SCR_004853)

    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 Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    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).

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore is not a substitute for expert review. SciScore checks for the presence and correctness of RRIDs (research resource identifiers) in the manuscript, and detects sentences that appear to be missing RRIDs. SciScore also checks to make sure that rigor criteria are addressed by authors. It does this by detecting sentences that discuss criteria such as blinding or power analysis. SciScore does not guarantee that the rigor criteria that it detects are appropriate for the particular study. Instead it assists authors, editors, and reviewers by drawing attention to sections of the manuscript that contain or should contain various rigor criteria and key resources. For details on the results shown here, including references cited, please follow this link.

  3. Version published to 10.1101/2020.07.07.190967v1 on bioRxiv