Chitinase 3-like-1 is a Therapeutic Target That Mediates the Effects of Aging in COVID-19

  1. Suchitra Kamle
  2. Bing Ma
  3. Chuan Hua He
  4. Bedia Akosman
  5. Yang Zhou
  6. Chang Min Lee
  7. Wafik S. El-Deiry
  8. Kelsey Huntington
  9. Olin Liang
  10. Jason T. Machan
  11. Min-Jong Kang
  12. Hyeon Jun Shin
  13. Emiko Mizoguchi
  14. Chun Geun Lee
  15. Jack A. Elias

Reviewed by

Read the full article See related articles

Listed in

Log in to save this article

Abstract

COVID-19 is caused by the SARS-CoV-2 (SC2) virus and is more prevalent and severe in the elderly and patients with comorbid diseases (CM). Because chitinase 3-like-1 (CHI3L1) is induced during aging and CM, the relationships between CHI3L1 and SC2 were investigated. Here we demonstrate that CHI3L1 is a potent stimulator of the SC2 receptor ACE2 and viral spike protein priming proteases (SPP), that ACE2 and SPP are induced during aging and that anti-CHI3L1, kasugamycin and inhibitors of phosphorylation, abrogate these ACE2- and SPP-inductive events. Human studies also demonstrated that the levels of circulating CHI3L1 are increased in the elderly and patients with CM where they correlate with COVID-19 severity. These studies demonstrate that CHI3L1 is a potent stimulator of ACE2 and SPP; that this induction is a major mechanism contributing to the effects of aging during SC2 infection and that CHI3L1 coopts the CHI3L1 axis to augment SC2 infection. CHI3L1 plays a critical role in the pathogenesis of and is an attractive therapeutic target in COVID-19.

Article activity feed

  1. Version published to 10.1101/2021.01.05.425478v2 on bioRxiv
  2. ScreenIT

    SciScore for 10.1101/2021.01.05.425478: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: The protocols that were used in these studies were evaluated and approved by the Institutional Animal Care and Use Committee (IACUC) at Brown University.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Th primary antibodies used in this study are: α-ACE2 (Abcam, ERR4435(2), ab108252), α-TMPRSS2 (Abcam, EPR3861, ab92323), α-CTSL (R&D Systems, AF1515). β-actin (Santa Cruz Biotechnology, Sc47778).
    α-ACE2
    suggested: None
    β-actin
    suggested: (Santa Cruz Biotechnology Cat# sc-47778, RRID:AB_626632)
    The sections were then incubated with primary antibodies (α-Ace2 (R&D Systems, AF3437), α-Tmprss2 (Abcam, ab92323), α-Ctsl (R&D Systems, AF1515), α-CC10 (Santa Cruz Biotechnology, sc-365992), α-SPC (Abcam, ab90716), α-CD31 (BD Pharmingen, 550274), α-transgelin (Abcam, ab14106)) overnight at 4°C.
    α-Tmprss2
    suggested: None
    α-Ctsl
    suggested: None
    α-CC10
    suggested: None
    α-SPC
    suggested: None
    α-CD31
    suggested: None
    α-transgelin
    suggested: None
    Double label immunohistochemistry: Double label immunohistochemistry was employed as previously described by our lab (34) Generation of monoclonal antibodies against CHI3L1 (FRG): The murine monoclonal anti-CHI3L1 antibody (FRG) was generated using peptide antigen (amino acid 223-234 of human CHI3L1) as immunogen through Abmart Inc (Berkeley Heights, NJ).
    CHI3L1
    suggested: None
    The lysates were then incubated with recombinant S protein for 2 hrs and the un-cleaved S and cleaved S1 and S2 proteins were evaluated by Western immunoblot analysis using anti-SARS-CoV-2 Spike antibody (Proscience, #3525).
    anti-SARS-CoV-2
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    HEK-293T cells were transfected with the FRG construct using Lipofectamine™ 3000 (Invitrogen, # L3000015).
    HEK-293T
    suggested: None
    Assessment of the effects of CHI3L1 on S protein-ACE2 binding and S protein processing: The effects of CHI3L1 on the binding of ACE2 and S protein were evaluated using Calu-3 cells and recombinant S protein.
    Calu-3
    suggested: KCLB Cat# 30055, RRID:CVCL_0609)
    The plasmids containing WT and 4 mutant forms of CHI3L1 (M230 (Ser-Arg), M235 (Ser-Arg), M237 (Thr-Phe), M239 (Tyr-Arg)) were transfected to HEK293 cells (ATCC® CRL-1573™) and each recombinant protein was purified using HisPur™ Ni-NTA column (Invitrogen, Cat# 88226).
    HEK293
    suggested: None
    Software and Algorithms
    SentencesResources
    Flow cytometry analysis of GFP (+) cells was carried out 48 h after infection on a BD LSRII flow cytometer and with the FlowJo software.
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Quantification and Statistical analysis: Statistical evaluations were undertaken with SPSS or GraphPad Prism software.
    SPSS
    suggested: (SPSS, RRID:SCR_002865)
    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 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 33. 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.

    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.

  3. Version published to 10.1101/2021.01.05.425478v1 on bioRxiv