Synergistic Block of SARS-CoV-2 Infection by Combined Drug Inhibition of the Host Entry Factors PIKfyve Kinase and TMPRSS2 Protease
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Abstract
Infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is causing the coronavirus disease 2019 (COVID-2019) global pandemic. There are ongoing efforts to uncover effective antiviral agents that could mitigate the severity of the disease by controlling the ensuing viral replication.
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SciScore for 10.1101/2021.06.01.446623: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Field Sample Permit: Isolation and propagation of SARS-CoV-2: Human samples were obtained under the Helsinki University Hospital laboratory research permit 30 HUS/32/2018 § 16. Isolation of SARS-CoV-2 from a COVID-19 Briefly, a nasopharyngeal swab in 500 μl of Copan UTM® Universal Transport Medium was inoculated on Calu-3 cells (P1) and incubated for 1 h at 37°C, after which the inoculum was removed and replaced with Minimum Essential Medium supplemented with 2% FBS, L-glutamine, penicillin and streptomycin. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication Contamination: The media was changed in all cell … SciScore for 10.1101/2021.06.01.446623: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Field Sample Permit: Isolation and propagation of SARS-CoV-2: Human samples were obtained under the Helsinki University Hospital laboratory research permit 30 HUS/32/2018 § 16. Isolation of SARS-CoV-2 from a COVID-19 Briefly, a nasopharyngeal swab in 500 μl of Copan UTM® Universal Transport Medium was inoculated on Calu-3 cells (P1) and incubated for 1 h at 37°C, after which the inoculum was removed and replaced with Minimum Essential Medium supplemented with 2% FBS, L-glutamine, penicillin and streptomycin. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication Contamination: The media was changed in all cell types every 2 days and regularly tested for presence of mycoplasma. Table 2: Resources
Antibodies Sentences Resources H6021, Sigma-Aldrich), and Alexa 647 fluorescently labelled goat anti-rabbit antibody (Thermo Fisher, cat. A32733) anti-rabbitsuggested: (Thermo Fisher Scientific Cat# A32733, RRID:AB_2633282)Briefly, viral NP was detected with an in house developed rabbit polyclonal antibody (31) counterstained with Alexa-647-conjugated goat anti rabbit secondary antibody, and nuclear staining done using Hoechst DNA dye. anti rabbit secondary antibody,suggested: NoneExperimental Models: Cell Lines Sentences Resources Cell Culture: VeroE6 (ATCC CRL-1586) VeroE6suggested: NoneVeroE6, VeroE6+TMPRSS2, Caco-2, and Calu-3 cells were maintained in DMEM supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin. Calu-3suggested: NoneBSR-T7 cells were derived from BHC cells (42) and grown in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin. BSR-T7suggested: CCLV Cat# CCLV-RIE 0583, RRID:CVCL_RW96)Caco-2 cells stably expressing human ACE2 were generated by transduction with third generation lentivirus pLenti7.3/V5 DEST ACE2-EmGFP (prepared by the cloning facility Dream-Lab, Institute of Biotechnology, University of Helsinki, Helsinki, Finland; the expression of Emerald GFP is driven by the SV40 promoter. Caco-2suggested: NoneIn all experiment’s cells were kept at 37oC and 5% (for Vero, Vero+TMRPSS2, and Caco-2 cells) or 7% CO2 (for Calu-3 cells) and media was pre warmed to 37°C. Verosuggested: NoneVirus titers were determined by plaque assay in VeroE6+TMPRSS2 cells. VeroE6+TMPRSS2suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)Software and Algorithms Sentences Resources For statistical analysis of the data in Figures 2 B, C and Figure 4 each replicate titration was fitted separately using IgorPro to determine the EC550 for each experiment. IgorProsuggested: NoneResults 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:
Identifier Status Title NCT04446377 Recruiting A Study of LAM-002A for the Prevention of Progression of COV… NCT04608266 Recruiting CAMOVID : Evaluation of Efficacy and Safety of Camostat Mesy… NCT04623021 Completed A Study Evaluating the Efficacy and Safety of CKD-314 (Nafab… NCT04418128 Not yet recruiting Clinical Efficacy of Nafamostat Mesylate for COVID-19 Pneumo… 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.
Results from scite Reference Check: We found no unreliable references.
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