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, 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.

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Published in eLife, v. 10.

© 2021, Fan et al.

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

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Funding Information

University of Louisville (1P01CA163223-01A1): Andrew N Lane; Teresa WM Fan

University of Louisville (1U24DK097215-01A1): Richard M Higashi; Teresa WM Fan; Andrew N Lane

University of Louisville (5P20GM121327): Andrew N Lane

Related Content

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 have been deposited at Dryad.

The following data sets were generated:

Lane AN Fan TWM Higashi RM Song H Daneshmandi S Mahan AL Purdom MS Pittman TA He D Wang C (2021) Dryad Digital Repository Innate immune activation by checkpoint inhibition in patient-derived lung cancer tissues. https://doi.org/10.5061/dryad.n5tb2rbwc

elife-69578-supp1-v4.docx (17 kB)
Transparent reporting form

elife-69578-supp2-v4.pdf (184 kB)
Supplementary file 1: Table S1. Patient demographics and clinical characteristics.

elife-69578-supp3-v4.pdf (213 kB)
Supplementary file 2: Primary NSCLC tissue of UK131 patient stains positive forsquamous cell carcinoma marker, PD-1 and PD-L1.

elife-69578-supp4-v4.docx (18 kB)
Supplementary file 3: Brain-metastasized NSCLC tissue of UK2035 patient stains positivefor neuroendocrine markers, PD-1 and PD-L1.

elife-69578-transrepform1-v4.docx (246 kB)
Supplementary file 4: Table S2. Information on primary antibodies used forimmunofluorescent staining.