Berberine and Obatoclax Inhibit SARS-Cov-2 Replication in Primary Human Nasal Epithelial Cells In Vitro
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Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged as a new human pathogen in late 2019 and it has infected over 100 million people in less than a year. There is a clear need for effective antiviral drugs to complement current preventive measures, including vaccines. In this study, we demonstrate that berberine and obatoclax, two broad-spectrum antiviral compounds, are effective against multiple isolates of SARS-CoV-2. Berberine, a plant-derived alkaloid, inhibited SARS-CoV-2 at low micromolar concentrations and obatoclax, which was originally developed as an anti-apoptotic protein antagonist, was effective at sub-micromolar concentrations. Time-of-addition studies indicated that berberine acts on the late stage of the viral life cycle. In agreement, berberine mildly affected viral RNA synthesis, but it strongly reduced infectious viral titers, leading to an increase in the particle-to-pfu ratio. In contrast, obatoclax acted at the early stage of the infection, which is in line with its activity to neutralize the acidic environment in endosomes. We assessed infection of primary human nasal epithelial cells that were cultured on an air-liquid interface and found that SARS-CoV-2 infection induced and repressed expression of specific sets of cytokines and chemokines. Moreover, both obatoclax and berberine inhibited SARS-CoV-2 replication in these primary target cells. We propose berberine and obatoclax as potential antiviral drugs against SARS-CoV-2 that could be considered for further efficacy testing.
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SciScore for 10.1101/2020.12.23.424189: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Post-operative residual tissue originating from the posterior nasal septum of a 17-year-old male was obtained after informed consent, according to the principles of the declaration of Helsinki.
IRB: The study was approved by the Committee on Research Involving Human Subjects Arnhem Nijmegen (CMO NL2020-6517) and conducted according to the principles of the Declaration of Helsinki (last updated 2013) and in accordance with the Medical Research Involving Human Subjects Act (Dutch: WMO).Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Post-operative residual tissue originating from the posterior nasal … SciScore for 10.1101/2020.12.23.424189: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Post-operative residual tissue originating from the posterior nasal septum of a 17-year-old male was obtained after informed consent, according to the principles of the declaration of Helsinki.
IRB: The study was approved by the Committee on Research Involving Human Subjects Arnhem Nijmegen (CMO NL2020-6517) and conducted according to the principles of the Declaration of Helsinki (last updated 2013) and in accordance with the Medical Research Involving Human Subjects Act (Dutch: WMO).Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Post-operative residual tissue originating from the posterior nasal septum of a 17-year-old male was obtained after informed consent, according to the principles of the declaration of Helsinki. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources S1) were stained for 1 h with anti-SARS-CoV Spike S1 subunit human IgG1 (BEI Resources) and mouse anti-dsRNA J2 monoclonal antibody (Scicons) anti-SARS-CoVsuggested: Nonehuman IgG1suggested: Noneanti-dsRNA J2suggested: (SCICONS Cat# 10010200, RRID:AB_2651015)Experimental Models: Cell Lines Sentences Resources Cells: African green monkey Vero E6 (ATCC CRL-1586) and Vero FM (ATCC CCL-81) kidney cells were grown in Dulbecco’s modified Eagle medium containing 4.5 g/L glucose and L-glutamine (Gibco), supplemented with 10% fetal calf serum (FCS, Sigma Aldrich), 100 μg/ml streptomycin and 100 U/ml penicillin (Gibco). E6suggested: NoneVerosuggested: NoneVero FM cells seeded in 24-well plates were infected in duplicate with 100 μl 2-fold serial dilutions of patient material in a total volume of 200 μl using virus isolation medium [DMEM (Gibco) containing, 20 mM HEPES buffer (Gibco), 100 μg/ml streptomycin (Gibco), 100 IU/ml penicillin (Gibco), 1% amphotericin B (Gibco)]. Vero FMsuggested: NoneVirus titration: Vero E6 cells were seeded onto 12-well plates at a density of 5 × 105 cells/well. Vero E6suggested: NoneSoftware and Algorithms Sentences Resources The 50% cytotoxicity concentration (CC50) value was estimated by four parameter logistic regression of the data, using Graphpad Prism (version 5.0). Graphpad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)S1B were captured using a Zeiss LSM9000 microscope with a 63× oil objective and processed using FIJI software. FIJIsuggested: (Fiji, RRID:SCR_002285)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:
Identifier Status Title NCT03281096 Unknown status A Research of Berberine Hydrochloride to Prevent Colorectal … NCT03333265 Recruiting Primary Chemoprevention of Familial Adenomatous Polyposis Wi… NCT03378934 Recruiting Anti-platelet Effect of Berberine in Patients After Percutan… NCT02808351 Recruiting Berberine Prevent Contrast-induced Nephropathy in Patients W… NCT02983188 Completed Berberine as Adjuvant Treatment for Schizophrenia Patients NCT03976336 Recruiting Evaluating the Tolerability and Effects of Berberine on Majo… NCT03198572 Recruiting Efficacy and Safety of Berberine in Non-alcoholic Steatohepa… NCT02737943 Unknown status Effect of Mebo Dressing Versus Standard Care on Managing Don… 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.
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