SARS-CoV-2 envelope protein causes acute respiratory distress syndrome (ARDS)-like pathological damages and constitutes an antiviral target
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
Cytokine storm and multi-organ failure are the main causes of SARS-CoV-2-related death. However, the origin of excessive damages caused by SARS-CoV-2 remains largely unknown. Here we show that the SARS-CoV-2 envelope (2-E) protein alone is able to cause acute respiratory distress syndrome (ARDS)-like damages in vitro and in vivo. 2-E proteins were found to form a type of pH-sensitive cation channels in bilayer lipid membranes. As observed in SARS-CoV-2-infected cells, heterologous expression of 2-E channels induced rapid cell death in various susceptible cell types and robust secretion of cytokines and chemokines in macrophages. Intravenous administration of purified 2-E protein into mice caused ARDS-like pathological damages in lung and spleen. A dominant negative mutation lowering 2-E channel activity attenuated cell death and SARS-CoV-2 production. Newly identified channel inhibitors exhibited potent anti-SARS-CoV-2 activity and excellent cell protective activity in vitro and these activities were positively correlated with inhibition of 2-E channel. Importantly, prophylactic and therapeutic administration of the channel inhibitor effectively reduced both the viral load and secretion of inflammation cytokines in lungs of SARS-CoV-2-infected transgenic mice expressing human angiotensin-converting enzyme 2 (hACE-2). Our study supports that 2-E is a promising drug target against SARS-CoV-2.
Article activity feed
-
-
SciScore for 10.1101/2020.06.27.174953: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: All animal procedures were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals, under protocols approved and strictly followed by the Institutional Animal Care and Use Committees (IACUC) Randomization All image data shown are representative of at least three randomly selected fields. Blinding not detected. Power Analysis not detected. Sex as a biological variable Male C57BL/6 mice were obtained from Shanghai SLAC Laboratory Animal Co., Ltd. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Blots were probed with antibodies against HA-Tag (C29F4) (3724, CST, USA), GSDMD … SciScore for 10.1101/2020.06.27.174953: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: All animal procedures were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals, under protocols approved and strictly followed by the Institutional Animal Care and Use Committees (IACUC) Randomization All image data shown are representative of at least three randomly selected fields. Blinding not detected. Power Analysis not detected. Sex as a biological variable Male C57BL/6 mice were obtained from Shanghai SLAC Laboratory Animal Co., Ltd. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Blots were probed with antibodies against HA-Tag (C29F4) (3724, CST, USA), GSDMD (L60) (93709, CST, USA), HA-Tagsuggested: (Cell Signaling Technology Cat# 3724, RRID:AB_1549585)Experimental Models: Cell Lines Sentences Resources Cell culture and treatment: HEK293, MCF-7, Caco2, Vero E6, HeLa, HepG2, SH-SY5Y cells were grown in 90% DMEM basal medium (Gibco, USA) supplemented with 10% fetal bovine serum (Gibco, USA), 2 mM L-glutamine and 100 units /mL penicillin/streptomycin (Gibco, USA). MCF-7suggested: NoneCaco2suggested: NoneVero E6suggested: RRID:CVCL_XD71)HeLasuggested: NoneHepG2suggested: NoneSH-SY5Ysuggested: None1% NEAA (Gibco, USA) was added in above medium for A498 cells culture. A498suggested: NCI-DTP Cat# A498, RRID:CVCL_1056)Besides, HCT116 and HT-29 cells were grown in McCoy’s 5A basal medium (Gibco, USA), PC3 and A549 cells were grown in RPMI-1640 basal medium (Hyclone, USA) and CHO cells were grown in DMEM/F-12 basal medium (Gibco, USA) supplemented as above. 16HBE cells were grown in KM (ScienCell, USA) medium. HCT116suggested: NoneHT-29suggested: NonePC3suggested: NoneA549suggested: NoneCHOsuggested: NoneExperimental Models: Organisms/Strains Sentences Resources Male C57BL/6 mice were obtained from Shanghai SLAC Laboratory Animal Co., Ltd. C57BL/6suggested: NoneSoftware and Algorithms Sentences Resources The percentages of differently labeled cells were calculated by FlowJo 7.6. FlowJosuggested: (FlowJo, RRID:SCR_008520)The MS raw data were analyzed with MaxQuant (http://maxquant.org/, MaxQuantsuggested: (MaxQuant, RRID:SCR_014485)The currents were digitized using pClamp 10.2 software (Molecular Devices, US). pClampsuggested: (pClamp, RRID:SCR_011323)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: We detected the following sentences addressing limitations in the study:One limitation of our study is the lack of evidence in the context of SARS-CoV-2 infection in vivo, which can technically be achieved by generating of SARS-CoV-2 without 2-E using reverse genetics. However, the 2-E deleted SARS-CoV-2 may replicate more effectively and thus this experiment was not conducted. Although the in vivo antiviral activity of the newly identified channel inhibitors needs further studies, their potent antiviral activity in vitro and excellent protection effects against ARDS-like damage in vivo shed light on the drug development of 2-E channel inhibitors. Given that 2-E can function as ion channels in the viral membranes, similar to how they function in host cells, we propose that 2-E channels may represent a new class of dualfunction targets against SARS-CoV-2.
Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 21. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.
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.
-
