Cholesterol 25-hydroxylase suppresses SARS-CoV-2 replication by blocking membrane fusion
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Abstract
The novel severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the etiological agent of coronavirus disease-2019 (COVID-19), has swept the world in unprecedented speed. In a few months, SARS-CoV-2 has infected millions of people and caused tens of thousands of deaths. There are no Food and Drug Administration-approved antivirals or vaccines yet available and clinical treatments are limited to supportive therapies that help alleviate the symptoms. Thus, there is an urgent need to identify effective antivirals as countermeasures before safe and effective vaccines are developed, tested, and then produced on a large scale. Our approach is to harness the germline-encoded interferon antiviral response to inhibit SARS-CoV-2 replication thereby limiting its pathogenicity.
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SciScore for 10.1101/2020.06.08.141077: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Cells were permeabilized and stained with antibodies against DAPI (P36962, Thermo Fisher), LAMP1 (9091S, Cell Signaling), LBPA (MABT837, Sigma), and Rab4 (ab13252, Abcam). antibodies against DAPI (P36962suggested: NoneLAMP1suggested: (Cell Signaling Technology Cat# 9091, RRID:AB_2687579)Rab4suggested: (Abcam Cat# ab13252, RRID:AB_2269374)Proteins were resolved in SDS-PAGE and analyzed by antibody as described (45) using the following antibodies and … SciScore for 10.1101/2020.06.08.141077: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Cells were permeabilized and stained with antibodies against DAPI (P36962, Thermo Fisher), LAMP1 (9091S, Cell Signaling), LBPA (MABT837, Sigma), and Rab4 (ab13252, Abcam). antibodies against DAPI (P36962suggested: NoneLAMP1suggested: (Cell Signaling Technology Cat# 9091, RRID:AB_2687579)Rab4suggested: (Abcam Cat# ab13252, RRID:AB_2269374)Proteins were resolved in SDS-PAGE and analyzed by antibody as described (45) using the following antibodies and dilutions: GAPDH (631402, Biolegend), GFP (2555S, Cell Signaling), SARS-CoV-2 S1 (40590-T62, Sino Biological), SARS-CoV-2 S2 (40592-T62, Sino Biological), and V5 (13202S, Cell Signaling). GAPDHsuggested: (BioLegend Cat# 631402, RRID:AB_2107422)GFPsuggested: (Cell Signaling Technology Cat# 2555, RRID:AB_10692764)V5 (13202S, Cell Signaling).suggested: NoneSecondary antibodies were anti-rabbit (#7074, Cell Signaling) or anti-mouse (#7076, Cell Signaling) immunoglobulin G horseradish peroxidase-linked antibodies. anti-rabbit (#7074, Cell Signaling)suggested: (Cell Signaling Technology Cat# 7074, RRID:AB_2099233)anti-mousesuggested: (Cell Signaling Technology Cat# 7076, RRID:AB_330924)Experimental Models: Cell Lines Sentences Resources Flow cytometry: HEK293-hACE2 or HEK293-hACE2-TMPRSS2 cells with or without CH25H expression were inoculated with wild-type VSV-GFP, VSV-SARS-CoV, or VSV-SARS-CoV-2 at an MOI = 10 (based on titers in Vero cells) for 1 hr at 37°C. HEK293-hACE2-TMPRSS2suggested: NoneVerosuggested: NoneSoftware and Algorithms Sentences Resources GFP positive cells were determined by BD LSRFortessa™ X-20 cell analyzer and analyzed by FlowJo v10.6.2 (BD). FlowJosuggested: (FlowJo, RRID:SCR_008520)Images were visualized by Volocity v6.3 and quantification was determined by ImageJ (NIH). Volocitysuggested: (Volocity 3D Image Analysis Software, RRID:SCR_002668)ImageJsuggested: (ImageJ, RRID:SCR_003070)1E, 3B, S1C, S1D, and S2B was calculated by Student’s t test using Prism 8.4.2 (GraphPad). Prismsuggested: (PRISM, RRID:SCR_005375)GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: Thank you for sharing your data.
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.
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