Anticancer pan-ErbB inhibitors reduce inflammation and tissue injury and exert broad-spectrum antiviral effects

  1. Sirle Saul
  2. Marwah Karim
  3. Luca Ghita
  4. Pei-Tzu Huang
  5. Winston Chiu
  6. Verónica Durán
  7. Chieh-Wen Lo
  8. Sathish Kumar
  9. Nishank Bhalla
  10. Pieter Leyssen
  11. Farhang Alem
  12. Niloufar A. Boghdeh
  13. Do H.N. Tran
  14. Courtney A. Cohen
  15. Jacquelyn A. Brown
  16. Kathleen E. Huie
  17. Courtney Tindle
  18. Mamdouh Sibai
  19. Chengjin Ye
  20. Ahmed Magdy Khalil
  21. Kevin Chiem
  22. Luis Martinez-Sobrido
  23. John M. Dye
  24. Benjamin A. Pinsky
  25. Pradipta Ghosh
  26. Soumita Das
  27. David E. Solow-Cordero
  28. Jing Jin
  29. John P. Wikswo
  30. Dirk Jochmans
  31. Johan Neyts
  32. Steven De Jonghe
  33. Aarthi Narayanan
  34. Shirit Einav

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Abstract

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  1. Version published to 10.1172/jci169510
  2. Version published to 10.1101/2021.05.15.444128v3 on bioRxiv
  3. Version published to 10.1101/2021.05.15.444128v2 on bioRxiv
  4. ScreenIT

    SciScore for 10.1101/2021.05.15.444128: (What is this?)

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

    Table 1: Rigor

    EthicsField Sample Permit: EBOV and MARV work was conducted at the high-containment BSL4 facilities at the United States Army Medical Research Institute of Infectious Diseases.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line AuthenticationContamination: All cells were tested negative for mycoplasma by the MycoAlert mycoplasma detection kit (Lonza, Morristown, NJ)

    Table 2: Resources

    Antibodies
    SentencesResources
    Blots were blocked with 5% BSA/TBST and blotted with anti-ErbB4 (Santa Cruz), ErbB2 (Cell Signaling), ErbB1, AKT, ERK, p38 (Cell Signaling), P-ErbB4 (Tyr1284), P-ErbB2(Tyr1248), P-AKT(Ser473), P-ERK(Thr202/Tyr204), P-p38 (Thr180/Tyr182) (Cell Signaling), and β-actin (Sigma-Aldrich, catalog A3854) antibodies.
    anti-ErbB4
    suggested: (Cell Signaling Technology Cat# 4757, RRID:AB_2099987)
    ErbB2
    suggested: None
    ERK
    suggested: (Thermo Fisher Scientific Cat# 85-86195-11, RRID:AB_2574775)
    p38
    suggested: None
    Tyr1284
    suggested: None
    Tyr1248
    suggested: None
    P-ERK(Thr202/Tyr204
    suggested: None
    β-actin ( Sigma-Aldrich , catalog A3854 ) antibodies
    suggested: None
    β-actin
    suggested: None
    Cell lysates were obtained at 1.5 and/or 24 hours post-infection followed by Western blot analysis with antibodies targeting the phosphorylated and total forms of ErbB1, 2 and 4, p38/MAPK, ERK, and AKT, as described(Zona et al., 2013).
    AKT
    suggested: None
    Cells were incubated with mouse mAb SARS-CoV-2 nucleocapsid antibody (SinoBiological, 1:100) and rabbit Claudin 7 polyclonal antibody (ThermoFisher, 1:200) overnight at 4°C, followed by 1 hour incubation at room temperature with goat anti-mouse AF488 (ThermoFisher, 1:400)
    anti-mouse
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Briefly, 30 µL Vero-E6-eGFP cells were added at 8000 cells/well to columns 1-24 24 hours before infection.
    Vero-E6-eGFP
    suggested: None
    Viral stocks preparation and/or passaging: Belgium-GHB-03021 SARS-CoV-2 strain was recovered from a nasopharyngeal swab taken from an asymptomatic patient returning from Wuhan, China early February 2020 (Spiteri et al., 2020) and passaged 6 times on Huh7 and Vero E6 cells.
    Vero E6
    suggested: RRID:CVCL_XD71)
    VEEV-TC-83-nLuc RNA was transcribed in vitro from cDNA plasmid templates linearized with MluI via MEGAscript SP6 kit (Invitrogen #AM1330) and electroporated into BHK-21 cells.
    BHK-21
    suggested: ATCC Cat# CRL-6282, RRID:CVCL_1914)
    Dose response curves with lapatinib in Vero and Calu-3 cells (Figures 1E and 2B, respectively) were performed with a P3 USA-WA1/2020 SARS-CoV-2 virus harboring no deletion or point mutations in the MBC domain.
    Calu-3
    suggested: KCLB Cat# 30055, RRID:CVCL_0609)
    Vero (ATCC), HEK-293T (ATCC)
    Vero
    suggested: None
    HEK-293T
    suggested: CCLV Cat# CCLV-RIE 1018, RRID:CVCL_0063)
    Huh7 cells (Apath LLC) and Calu-3 cells (ATCC) were grown in DMEM (10-013-CV, Corning) supplemented with 10% FBS (Omega Scinetific, Inc), 1% l-glutamine, 1% nonessential amino acids (Corning) and 1% penicillin-streptomycin (Gibco).
    Huh7
    suggested: None
    Resistance studies: VEEV (TC-83) was used to inoculate U-87 MG cells at MOI of 0.1 and passaged every 24 hours by transferring an equal volume of viral supernatant to naive cells under increasing drug selection (2.5-5 μM, passages 1–3; 5-10 μM, passages 4-7; 10-15 μM, passages 8-10).
    U-87 MG
    suggested: None
    Recombinant DNA
    SentencesResources
    DENV2 (New Guinea C strain) TSV01 Renilla reporter plasmid (pACYC NGC FL) was a gift from Pei-Yong Shi (University of Texas Medical Branch, Galveston, Texas, USA) (Zou et al., 2011).
    pACYC
    suggested: RRID:Addgene_102793)
    pDONR223-ErbB4 was a gift from William Hahn & David Root (Addgene plasmid # 23875; http://n2t.net/addgene:23875; RRID:Addgene_23875) (Johannessen et al., 2010).
    detected: RRID:Addgene_23875)
    The ORF was recombined into a gateway compatible pGluc destination vector using Gateway technology (Invitrogen) and the construct was verified using Sanger sequencing.
    pGluc
    suggested: None
    Similarly, DENV RNA was transcribed in vitro from pACYC-DENV2-NGC plasmid by mMessage/mMachine (
    pACYC-DENV2-NGC
    suggested: None
    Software and Algorithms
    SentencesResources
    Band intensity was quantified with ImageJ software (NIH)
    ImageJ
    suggested: (ImageJ, RRID:SCR_003070)
    Adjustment for brightness, contrast and color balance were done using Fiji software.
    Fiji
    suggested: (Fiji, RRID:SCR_002285)
    Quantification and Statistical Analysis: All data were analyzed with GraphPad Prism software.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    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.

    Results from scite Reference Check: We found no unreliable references.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  5. Version published to 10.1101/2021.05.15.444128v1 on bioRxiv