Hydroxychloroquine (HCQ) decreases the benefit of anti-PD-1 immune checkpoint blockade in tumor immunotherapy

  1. Janna Krueger
  2. Francois Santinon
  3. Alexandra Kazanova
  4. Mark E. Issa
  5. Bruno Larrivee
  6. Richard Kremer
  7. Catalin Milhalcioiu
  8. Christopher E. Rudd

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Abstract

Immunotherapy using checkpoint blockade (ICB) with antibodies such as anti-PD-1 has revolutionised the treatment of many cancers. Despite its use to treat COVID-19 patients and autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis, the effect of hydroxychloroquine (HCQ) on cancer immunotherapy has not been examined. In this study, remarkably, we find that HCQ alone, or in combination with azithromycin (AZ), at doses used to treat patients, decreased the therapeutic benefit of anti-PD-1 in cancer immunotherapy. No deleterious effect was seen on untreated tumors. Mechanistically, HCQ and HCQ/AZ inhibited PD-L1 expression on tumor cells, while specifically targeting the anti-PD-1 induced increase in progenitor CD8 + CD44 + PD-1 + TCF1 + tumor infiltrating T cells (TILs) and the generation of CD8 + CD44 + PD-1 + effectors. Surprisingly, it also impaired the appearance of a subset of terminally exhausted CD8 + TILs. No effect was seen on the presence of CD4 + T cells, FoxP3 + regulatory T cells (Tregs), thymic subsets, B cells, antibody production, myeloid cells, or the vasculature of mice. This study indicates for the first time that HCQ and HCQ/AZ negatively impact the ability of anti-PD-1 checkpoint blockade to promote tumor rejection.

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  1. Version published to 10.1371/journal.pone.0251731
  2. ScreenIT

    SciScore for 10.1101/2020.09.29.20193110: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: The human study was approved by the McGill University Health Centre (MUHC) Research Ethics Board #2021-6881.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Anti-B16-PDL1 antibody assay: Plasma was collected from heparinized murine peripheral blood by cetrifugation for 15 min at 2000g.
    Anti-B16-PDL1
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Mice, aged 7-8 weeks, were implanted intradermally with 50,000 B16-PD-L1 melanoma cells that overexpress PD-L1.
    B16-PD-L1
    suggested: None
    Briefly, 5×105 B16-PDL1 tumour cells were added to a 96-well plated, blocked with 10% FBS-PBS, and washed with PBS.
    B16-PDL1
    suggested: None
    Plasma samples were diluted 1:16 in PBS and were added to B16 cells in a volume of 100 μl, mixed and then incubated for 1 hour at 4°C.
    B16
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Mice: C57BL/6 mice were housed at the Hôpital Maisonneuve Rosemont animal facility (Montreal, QC, Canada).
    C57BL/6
    suggested: None
    Software and Algorithms
    SentencesResources
    FACS analyses were performed by using FlowJo software or Cytobank for viSNE analyses.
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Cytobank
    suggested: (Cytobank, RRID:SCR_014043)
    Quantification of retinal vasculature branch points was done with the use of the angiogenesis analyzer tool for ImageJ.
    ImageJ
    suggested: (ImageJ, RRID:SCR_003070)

    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: Please consider improving the rainbow (“jet”) colormap(s) used on pages 41, 36 and 38. 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 10.1101/2020.09.29.20193110v1 on medRxiv