Innate immune activation by checkpoint inhibition in human patient-derived lung cancer tissues

  1. Teresa Fan  Is a corresponding author
  2. Richard M Higashi
  3. Huan Song
  4. Saeed Daneshmandi
  5. Angela L Mahan
  6. Matthew S Purdom
  7. Therese J Bocklage
  8. Thomas A Pittman
  9. Daheng He
  10. Chi Wang
  11. Andrew N Lane  Is a corresponding author
  1. University of Kentucky, United States
  2. Baptist Health, United States

Abstract

Although Pembrolizumab-based immunotherapy has significantly improved lung cancer patient survival, many patients show variable efficacy and resistance development. A better understanding of the drug's action is needed to improve patient outcomes. Functional heterogeneity of the tumor microenvironment (TME) is crucial to modulating drug resistance; understanding of individual patients' TME that impacts drug response is hampered by lack of appropriate models. Lung organotypic tissue slice cultures (OTC) with patients' native TME procured from primary and brain-metastasized (BM) non-small cell lung cancer (NSCLC) patients were treated with Pembrolizumab and/or beta-glucan (WGP, an innate immune activator). Metabolic tracing with 13C6-Glc/13C5,15N2-Gln, multiplex immunofluorescence (mIF), and digital spatial profiling (DSP) were employed to interrogate metabolic and functional responses to Pembrolizumab and/or WGP. Primary and BM PD-1+ lung cancer OTC responded to Pembrolizumab and Pembrolizumab + WGP treatments, respectively. Pembrolizumab activated innate immune metabolism and functions in primary OTC, which were accompanied by tissue damage. DSP analysis indicated an overall decrease in immunosuppressive macrophages and T cells but revealed microheterogeneity in immune responses and tissue damage. Two TMEs with altered cancer cell properties showed resistance. Pembrolizumab or WGP alone had negligible effects on BM-lung cancer OTC but Pembrolizumab + WGP blocked central metabolism with increased pro-inflammatory effector release and tissue damage. In depth metabolic analysis and multiplex TME imaging of lung cancer OTC demonstrated overall innate immune activation by Pembrolizumab but heterogeneous responses in the native TME of a patient with primary NSCLC. Metabolic and functional analysis also revealed synergistic action of Pembrolizumab and WGP in OTC of metastatic NSCLC.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Excel spreadsheets of data used for tables and figures will be deposited at Dryad.

The following data sets were generated

Article and author information

Author details

  1. Teresa Fan

    University of Kentucky, LEXINGTON, United States
    For correspondence
    teresa.fan@uky.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7292-8938
  2. Richard M Higashi

    Department of Toxicology and Cancer Biology, University of Kentucky, Lexington, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Huan Song

    Department of Toxicology and Cancer Biology, University of Kentucky, Lexington, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Saeed Daneshmandi

    Department of Toxicology and Cancer Biology, University of Kentucky, Lexington, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Angela L Mahan

    Baptist Health, Louisville, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Matthew S Purdom

    Deptartment of Pathology and Laboratory Medicine, University of Kentucky, Lexington, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Therese J Bocklage

    Deptartment of Pathology and Laboratory Medicine, University of Kentucky, Lexington, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Thomas A Pittman

    Department of Neurosurgery, University of Kentucky, Lexington, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Daheng He

    Deptartment of Pathology and Laboratory Medicine, University of Kentucky, Lexington, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Chi Wang

    Deptartment of Pathology and Laboratory Medicine, University of Kentucky, Lexington, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Andrew N Lane

    Department of Toxicology and Cancer Biology, University of Kentucky, Lexington, United States
    For correspondence
    andrew.lane@uky.edu
    Competing interests
    The authors declare that no competing interests exist.

Funding

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Human subjects: Surgical patients were consented for freshly resected tissue specimens under the approved IRB protocol (14-0288-F6A; 13-LUN-94-MCC) of the University of Kentucky (UKy).

Reviewing Editor

  1. Fabio Malavasi, University of Torino Medical School, Italy

Publication history

  1. Received: April 20, 2021
  2. Accepted: July 26, 2021
  3. Accepted Manuscript published: August 18, 2021 (version 1)
  4. Accepted Manuscript updated: August 20, 2021 (version 2)
  5. Accepted Manuscript updated: August 31, 2021 (version 3)
  6. Version of Record published: September 27, 2021 (version 4)

Copyright

© 2021, Fan et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Teresa Fan
  2. Richard M Higashi
  3. Huan Song
  4. Saeed Daneshmandi
  5. Angela L Mahan
  6. Matthew S Purdom
  7. Therese J Bocklage
  8. Thomas A Pittman
  9. Daheng He
  10. Chi Wang
  11. Andrew N Lane
(2021)
Innate immune activation by checkpoint inhibition in human patient-derived lung cancer tissues
eLife 10:e69578.
https://doi.org/10.7554/eLife.69578
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    We searched the PubMed (MEDLINE) database for articles regarding sarcoma-associated ncRNAs from inception to August 17, 2022. Studies investigating the roles of host-derived miRNAs, long ncRNAs, and circular RNAs in sarcoma were included. Data relating to the roles of ncRNAs in therapeutic regulation and their applicability as biomarkers for predicting the therapeutic response of sarcomas were extracted. Two independent researchers assessed the quality of the studies using the Würzburg Methodological Quality Score (W-MeQS).

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    Observational studies revealed the ectopic expression of ncRNAs in sarcoma patients who had different responses to antitumor treatments. Experimental studies have confirmed crosstalk between cellular pathways pertinent to chemotherapy, targeted therapy, and radiotherapy resistance. Of the included studies, W-MeQS scores ranged from 3 to 10 (average score = 5.42). Of the 12 articles that investigated ncRNAs as biomarkers, none included a validation cohort. Selective reporting of the sensitivity, specificity, and receiver operating curves was common.

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    Although ncRNAs appear to be good candidates as biomarkers for predicting treatment response and therapeutics for sarcoma, their differential expression across tissues complicates their application. Further research regarding their potential for inhibiting or activating these regulatory molecules to reverse treatment resistance may be useful.

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