HTCC as a highly effective polymeric inhibitor of SARS-CoV-2 and MERS-CoV

  1. Aleksandra Milewska
  2. Ying Chi
  3. Artur Szczepanski
  4. Emilia Barreto-Duran
  5. Kevin Liu
  6. Dan Liu
  7. Xiling Guo
  8. Yiyue Ge
  9. Jingxin Li
  10. Lunbiao Cui
  11. Marek Ochman
  12. Maciej Urlik
  13. Sylwia Rodziewicz-Motowidlo
  14. Fengcai Zhu
  15. Krzysztof Szczubialka
  16. Maria Nowakowska
  17. Krzysztof Pyrc

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Abstract

The beginning of 2020 brought us information about the novel coronavirus emerging in China. Rapid research resulted in the characterization of the pathogen, which appeared to be a member of the SARS-like cluster, commonly seen in bats. Despite the global and local efforts, the virus escaped the healthcare measures and rapidly spread in China and later globally, officially causing a pandemic and global crisis in March 2020. At present, different scenarios are being written to contain the virus, but the development of novel anticoronavirals for all highly pathogenic coronaviruses remains the major challenge. Here, we describe the antiviral activity of previously developed by us HTCC compound (N-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride), which may be used as potential inhibitor of currently circulating highly pathogenic coronaviruses – SARS-CoV-2 and MERS-CoV.

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  1. ScreenIT

    SciScore for 10.1101/2020.03.29.014183: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: The study was approved by the Bioethical Committee of the Medical University of Silesia in Katowice, Poland (approval no: KNW/0022/KB1/17/10 dated 16.02.2010).
    Consent: Written consent was obtained from all patients.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Cell culture: Vero and Vero E6 (Cercopithecus aethiops; kidney epithelial; ATCC: CCL-81 and CRL-1586), Huh7 (Homo sapiens; hepatocellular carcinoma; ECACC: 01042712) and A549 cells with ACE2 overexpression (A549/ACE2)29 were cultured in Dulbecco’s MEM (Thermo Fisher Scientific, Poland) supplemented with 3% fetal bovine serum (heat-inactivated; Thermo Fisher Scientific, Poland) and antibiotics: penicillin (100 U/ml), streptomycin (100 μg/ml), and ciprofloxacin (5 μg/ml).
    A549
    suggested: None
    A549/ACE2)29
    suggested: None
    Virus preparation and titration: MERS-CoV stock (isolate England 1, 1409231v, National Collection of Pathogenic Viruses, Public Health England, United Kingdom) was generated by infecting monolayers of Vero cells.
    Vero
    suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)
    Virus yield was assessed by titration on fully confluent Vero or Vero E6 cells in 96-well plates, according to the method of Reed and Muench.
    Vero E6
    suggested: None
    Virus infection: In in vitro experiments, fully confluent Vero, Vero E6, or Huh7 cells in 96-well plates (TPP) were exposed to MERS-CoV, SARS-CoV-2 or mock at a TCID50 of 400 per ml in the presence of tested polymer or control medium.
    Huh7
    suggested: None
    Pseudovirus production and transduction: 293T cells were seeded on 10 cm2 dishes, cultured for 24 h at 37°C with 5% CO2 and transfected using polyethyleneimine (Sigma-Aldrich, Poland) with the lentiviral packaging plasmid (psPAX), the VSV-G envelope plasmid (pMD2G) or SARS-CoV-2 S glycoprotein (pCAGGS-SARS-CoV-2-S) and third plasmid encoding GFP protein (Lego-G2).
    293T
    suggested: None
    A549/ACE2 cells were seeded in 48-wells plates, cultured for 24 h at 37°C with 5% CO2 and transduced with pseudoviruses harboring VSV-G or S-SARS-CoV-2 proteins or lacking the fusion protein (∆Env) in the presence of polybrene (4 µg/ml; Sigma-Aldrich, Poland) and HTCC-77 (100 μg/ml) or control PBS.
    A549/ACE2
    suggested: None
    Software and Algorithms
    SentencesResources
    The Netherlands) and processed using ImageJ 1.47v (National Institutes of Health, Bethesda, MD, USA).
    ImageJ
    suggested: (ImageJ, RRID:SCR_003070)

    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

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  2. Version published to 10.1101/2020.03.29.014183v1 on bioRxiv