Journal Article
. 2021 Jul; 11:611365.
doi: 10.3389/fonc.2021.611365.

Explant Modeling of the Immune Environment of Head and Neck Cancer

Shay Sharon 1 Thomas Duhen 2 Shelly Bambina 2 Jason Baird 2 Rom Leidner 2 Bryan Bell 2 Nardy Casap 1 Marka Crittenden 2 Swetha Vasudevan 3 Maria Jubran 3 Nataly Kravchenko-Balasha 3 Michael Gough 2 
  • PMID: 34221953
  •     58 References


Patients exhibit distinct responses to immunotherapies that are thought to be linked to their tumor immune environment. However, wide variations in outcomes are also observed in patients with matched baseline tumor environments, indicating that the biological response to treatment is not currently predictable using a snapshot analysis. To investigate the relationship between the immune environment of tumors and the biological response to immunotherapies, we characterized four murine head and neck squamous cell carcinoma (HNSCC) models on two genetic backgrounds. Using tumor explants from those models, we identified correlations between the composition of infiltrating immune cells and baseline cytokine profiles prior to treatment. Following treatment with PD-1 blockade, CTLA-4 blockade, or OX40 stimulation, we observed inter-individual variability in the response to therapy between genetically identical animals bearing the same tumor. These distinct biological responses to treatment were not linked to the initial tumor immune environment, meaning that outcome would not be predictable from a baseline analysis of the tumor infiltrates. We similarly performed the explant assay on patient HNSCC tumors and found significant variability between the baseline environment of the tumors and their response to therapy. We propose that tumor explants provide a rapid biological assay to assess response to candidate immunotherapies that may allow matching therapies to individual patient tumors. Further development of explant approaches may allow screening and monitoring of treatment responses in HNSCC.

Keywords: CTLA4; OX40; PD1; cytokine; explant; head and neck cancer; immunotherapy; tumor.

IFN-γ-related mRNA profile predicts clinical response to PD-1 blockade.
Mark Ayers, Jared Lunceford, +12 authors, Terrill K McClanahan.
J Clin Invest, 2017 Jun 27; 127(8). PMID: 28650338    Free PMC article.
Highly Cited.
Dendritic Cell Maturation Defines Immunological Responsiveness of Tumors to Radiation Therapy.
Tiffany C Blair, Shelly Bambina, +8 authors, Michael J Gough.
J Immunol, 2020 Apr 29; 204(12). PMID: 32341058    Free PMC article.
The Society for Immunotherapy of Cancer consensus statement on immunotherapy for the treatment of squamous cell carcinoma of the head and neck (HNSCC).
Ezra E W Cohen, R Bryan Bell, +16 authors, Robert L Ferris.
J Immunother Cancer, 2019 Jul 17; 7(1). PMID: 31307547    Free PMC article.
Highly Cited.
LacZ inducible, antigen/MHC-specific T cell hybrids.
S Sanderson, N Shastri.
Int Immunol, 1994 Mar 01; 6(3). PMID: 8186188
Highly Cited.
Tumor vaccines expressing flt3 ligand synergize with ctla-4 blockade to reject preimplanted tumors.
Michael A Curran, James P Allison.
Cancer Res, 2009 Sep 10; 69(19). PMID: 19738077    Free PMC article.
Defining Immunogenic and Radioimmunogenic Tumors.
Terry R Medler, Tiffany C Blair, Marka R Crittenden, Michael J Gough.
Front Oncol, 2021 Apr 06; 11. PMID: 33816320    Free PMC article.
The neoadjuvant paradigm reinvigorated: a review of pre-surgical immunotherapy in HNSCC.
Margaret Stafford, John Kaczmar.
Cancers Head Neck, 2020 Mar 21; 5. PMID: 32195008    Free PMC article.
Fc-dependent depletion of tumor-infiltrating regulatory T cells co-defines the efficacy of anti-CTLA-4 therapy against melanoma.
Tyler R Simpson, Fubin Li, +11 authors, Sergio A Quezada.
J Exp Med, 2013 Jul 31; 210(9). PMID: 23897981    Free PMC article.
Highly Cited.
Digital pathology-aided assessment of tumor-infiltrating T lymphocytes in advanced stage, HPV-negative head and neck tumors.
Emma J de Ruiter, Reinout H de Roest, +4 authors, Stefan M Willems.
Cancer Immunol Immunother, 2020 Jan 26; 69(4). PMID: 31980916    Free PMC article.
Evaluation of Explant Responses to STING Ligands: Personalized Immunosurgical Therapy for Head and Neck Squamous Cell Carcinoma.
Jason R Baird, R Bryan Bell, +12 authors, Michael J Gough.
Cancer Res, 2018 Sep 19; 78(21). PMID: 30224374
Mechanism of tumor rejection with doublets of CTLA-4, PD-1/PD-L1, or IDO blockade involves restored IL-2 production and proliferation of CD8(+) T cells directly within the tumor microenvironment.
Stefani Spranger, Holly K Koblish, +3 authors, Thomas F Gajewski.
J Immunother Cancer, 2014 May 16; 2. PMID: 24829760    Free PMC article.
Highly Cited.
Bystander CD8+ T cells are abundant and phenotypically distinct in human tumour infiltrates.
Yannick Simoni, Etienne Becht, +29 authors, Evan W Newell.
Nature, 2018 May 18; 557(7706). PMID: 29769722
Highly Cited.
The prognostic role of tumor infiltrating T-lymphocytes in squamous cell carcinoma of the head and neck: A systematic review and meta-analysis.
Emma J de Ruiter, Marc L Ooft, Lot A Devriese, Stefan M Willems.
Oncoimmunology, 2017 Nov 18; 6(11). PMID: 29147608    Free PMC article.
Highly Cited. Review.
Decoding cancer heterogeneity: studying patient-specific signaling signatures towards personalized cancer therapy.
Efrat Flashner-Abramson, Swetha Vasudevan, +2 authors, Nataly Kravchenko-Balasha.
Theranostics, 2019 Aug 15; 9(18). PMID: 31410207    Free PMC article.
Personalized disease signatures through information-theoretic compaction of big cancer data.
Swetha Vasudevan, Efrat Flashner-Abramson, +2 authors, Nataly Kravchenko-Balasha.
Proc Natl Acad Sci U S A, 2018 Jul 07; 115(30). PMID: 29976841    Free PMC article.
Genetic basis for clinical response to CTLA-4 blockade in melanoma.
Alexandra Snyder, Vladimir Makarov, +18 authors, Timothy A Chan.
N Engl J Med, 2014 Nov 20; 371(23). PMID: 25409260    Free PMC article.
Highly Cited.
Cancer immunology. Mutational landscape determines sensitivity to PD-1 blockade in non-small cell lung cancer.
Naiyer A Rizvi, Matthew D Hellmann, +21 authors, Timothy A Chan.
Science, 2015 Mar 15; 348(6230). PMID: 25765070    Free PMC article.
Highly Cited.
Genomic correlates of response to CTLA-4 blockade in metastatic melanoma.
Eliezer M Van Allen, Diana Miao, +19 authors, Levi A Garraway.
Science, 2015 Sep 12; 350(6257). PMID: 26359337    Free PMC article.
Highly Cited.
International validation of the consensus Immunoscore for the classification of colon cancer: a prognostic and accuracy study.
Franck Pagès, Bernhard Mlecnik, +96 authors, Jérôme Galon.
Lancet, 2018 May 15; 391(10135). PMID: 29754777
Highly Cited.
CD8+ tumour-infiltrating lymphocytes in relation to HPV status and clinical outcome in patients with head and neck cancer after postoperative chemoradiotherapy: A multicentre study of the German cancer consortium radiation oncology group (DKTK-ROG).
Panagiotis Balermpas, Franz Rödel, +21 authors, Emmanouil Fokas.
Int J Cancer, 2015 Jul 17; 138(1). PMID: 26178914
Type, density, and location of immune cells within human colorectal tumors predict clinical outcome.
Jérôme Galon, Anne Costes, +13 authors, Franck Pagès.
Science, 2006 Sep 30; 313(5795). PMID: 17008531
Highly Cited.
PD-1 and PD-L1 expression in HNSCC primary cancer and related lymph node metastasis - impact on clinical outcome.
Sven Schneider, Lorenz Kadletz, +7 authors, Gregor Heiduschka.
Histopathology, 2018 May 10; 73(4). PMID: 29742291
Anti-CTLA-4 antibodies of IgG2a isotype enhance antitumor activity through reduction of intratumoral regulatory T cells.
Mark J Selby, John J Engelhardt, +4 authors, Alan J Korman.
Cancer Immunol Res, 2014 Apr 30; 1(1). PMID: 24777248
Highly Cited.
Association between severe treatment-related lymphopenia and progression-free survival in patients with newly diagnosed squamous cell head and neck cancer.
Jian L Campian, Guneet Sarai, +2 authors, Stuart A Grossman.
Head Neck, 2013 Nov 01; 36(12). PMID: 24174270    Free PMC article.
PD-L1 expression in cancer patients receiving anti PD-1/PD-L1 antibodies: A systematic review and meta-analysis.
Sara Gandini, Daniela Massi, Mario Mandalà.
Crit Rev Oncol Hematol, 2016 Feb 21; 100. PMID: 26895815
Highly Cited. Systematic Review.
Association of Immunologic Markers With Survival in Upfront Resectable Pancreatic Cancer.
Ephraim S Tang, Philippa H Newell, +4 authors, Michael J Gough.
JAMA Surg, 2018 Aug 10; 153(11). PMID: 30090929    Free PMC article.
Tumor infiltrating lymphocytes and survival in patients with head and neck squamous cell carcinoma.
Nghia Nguyen, Emily Bellile, +15 authors, Head and Neck SPORE Program Investigators.
Head Neck, 2016 Feb 18; 38(7). PMID: 26879675    Free PMC article.
Highly Cited.
Association of PD-1, PD-1 ligands, and other features of the tumor immune microenvironment with response to anti-PD-1 therapy.
Janis M Taube, Alison Klein, +7 authors, Robert A Anders.
Clin Cancer Res, 2014 Apr 10; 20(19). PMID: 24714771    Free PMC article.
Highly Cited.
The Dynamic Entropy of Tumor Immune Infiltrates: The Impact of Recirculation, Antigen-Specific Interactions, and Retention on T Cells in Tumors.
Tiffany C Blair, Alejandro F Alice, +2 authors, Michael J Gough.
Front Oncol, 2021 May 11; 11. PMID: 33968757    Free PMC article.
PD-1 and CTLA-4 combination blockade expands infiltrating T cells and reduces regulatory T and myeloid cells within B16 melanoma tumors.
Michael A Curran, Welby Montalvo, Hideo Yagita, James P Allison.
Proc Natl Acad Sci U S A, 2010 Feb 18; 107(9). PMID: 20160101    Free PMC article.
Highly Cited.
Novel prognostic clinical factors and biomarkers for outcome prediction in head and neck cancer: a systematic review.
Volker Budach, Ingeborg Tinhofer.
Lancet Oncol, 2019 Jun 05; 20(6). PMID: 31162105
Systematic Review.
OX40 (CD134) controls memory T helper 2 cells that drive lung inflammation.
Shahram Salek-Ardakani, Jianxun Song, +4 authors, Michael Croft.
J Exp Med, 2003 Jul 16; 198(2). PMID: 12860930    Free PMC article.
Exploring Alzheimer's Disease Molecular Variability via Calculation of Personalized Transcriptional Signatures.
Hila Dagan, Efrat Flashner-Abramson, +3 authors, Nataly Kravchenko-Balasha.
Biomolecules, 2020 Apr 01; 10(4). PMID: 32225014    Free PMC article.
ERK1/2 regulation of CD44 modulates oral cancer aggressiveness.
Nancy P Judd, Ashley E Winkler, +7 authors, Ravindra Uppaluri.
Cancer Res, 2011 Nov 17; 72(1). PMID: 22086849    Free PMC article.
Co-expression of CD39 and CD103 identifies tumor-reactive CD8 T cells in human solid tumors.
Thomas Duhen, Rebekka Duhen, +12 authors, Andrew D Weinberg.
Nat Commun, 2018 Jul 15; 9(1). PMID: 30006565    Free PMC article.
Highly Cited.
PD-1 blockade augments Th1 and Th17 and suppresses Th2 responses in peripheral blood from patients with prostate and advanced melanoma cancer.
John Dulos, Gregory J Carven, +12 authors, Annemieke M Boots.
J Immunother, 2012 Feb 07; 35(2). PMID: 22306905
QuPath: Open source software for digital pathology image analysis.
Peter Bankhead, Maurice B Loughrey, +10 authors, Peter W Hamilton.
Sci Rep, 2017 Dec 06; 7(1). PMID: 29203879    Free PMC article.
Highly Cited.
Predictive correlates of response to the anti-PD-L1 antibody MPDL3280A in cancer patients.
Roy S Herbst, Jean-Charles Soria, +19 authors, F Stephen Hodi.
Nature, 2014 Nov 28; 515(7528). PMID: 25428504    Free PMC article.
Highly Cited.
Combination CTLA-4 blockade and 4-1BB activation enhances tumor rejection by increasing T-cell infiltration, proliferation, and cytokine production.
Michael A Curran, Myoungjoo Kim, +2 authors, James P Allison.
PLoS One, 2011 May 12; 6(4). PMID: 21559358    Free PMC article.
Highly Cited.
Pembrolizumab for the treatment of non-small-cell lung cancer.
Edward B Garon, Naiyer A Rizvi, +25 authors, KEYNOTE-001 Investigators.
N Engl J Med, 2015 Apr 22; 372(21). PMID: 25891174
Highly Cited.
Pan-cancer deconvolution of tumour composition using DNA methylation.
Ankur Chakravarthy, Andrew Furness, +10 authors, Tim R Fenton.
Nat Commun, 2018 Aug 15; 9(1). PMID: 30104673    Free PMC article.
On a fundamental structure of gene networks in living cells.
Nataly Kravchenko-Balasha, Alexander Levitzki, +4 authors, R D Levine.
Proc Natl Acad Sci U S A, 2012 Mar 07; 109(12). PMID: 22392990    Free PMC article.
ClustVis: a web tool for visualizing clustering of multivariate data using Principal Component Analysis and heatmap.
Tauno Metsalu, Jaak Vilo.
Nucleic Acids Res, 2015 May 15; 43(W1). PMID: 25969447    Free PMC article.
Highly Cited.
Timing of PD-1 Blockade Is Critical to Effective Combination Immunotherapy with Anti-OX40.
David J Messenheimer, Shawn M Jensen, +6 authors, Bernard A Fox.
Clin Cancer Res, 2017 Sep 01; 23(20). PMID: 28855348    Free PMC article.
Highly Cited.
Multispectral Imaging of T and B Cells in Murine Spleen and Tumor.
Zipei Feng, Shawn M Jensen, +4 authors, Bernard A Fox.
J Immunol, 2016 Mar 20; 196(9). PMID: 26994219    Free PMC article.
Tumor cure by radiation therapy and checkpoint inhibitors depends on pre-existing immunity.
Marka R Crittenden, Lauren Zebertavage, +7 authors, Michael J Gough.
Sci Rep, 2018 May 05; 8(1). PMID: 29725089    Free PMC article.
Multiparametric immune profiling in HPV- oral squamous cell cancer.
Zipei Feng, Daniel Bethmann, +14 authors, Bernard A Fox.
JCI Insight, 2017 Jul 21; 2(14). PMID: 28724788    Free PMC article.
Highly Cited.
PD-L1: a novel prognostic biomarker in head and neck squamous cell carcinoma.
Tim Müller, Martin Braun, +9 authors, Peter Brossart.
Oncotarget, 2017 Sep 09; 8(32). PMID: 28881780    Free PMC article.
Cutting Edge: OX40 agonists can drive regulatory T cell expansion if the cytokine milieu is right.
Carl E Ruby, Melissa A Yates, +5 authors, Andrew D Weinberg.
J Immunol, 2009 Sep 30; 183(8). PMID: 19786544    Free PMC article.
CTLA4 blockade and GM-CSF combination immunotherapy alters the intratumor balance of effector and regulatory T cells.
Sergio A Quezada, Karl S Peggs, Michael A Curran, James P Allison.
J Clin Invest, 2006 Jun 17; 116(7). PMID: 16778987    Free PMC article.
Highly Cited.
mTOR and MEK1/2 inhibition differentially modulate tumor growth and the immune microenvironment in syngeneic models of oral cavity cancer.
Harrison Cash, Sujay Shah, +4 authors, Clint Allen.
Oncotarget, 2015 Oct 28; 6(34). PMID: 26506415    Free PMC article.
The head and neck cancer immune landscape and its immunotherapeutic implications.
Rajarsi Mandal, Yasin Şenbabaoğlu, +8 authors, Luc Gt Morris.
JCI Insight, 2016 Oct 26; 1(17). PMID: 27777979    Free PMC article.
Highly Cited.
Cell cycle and viral and immunologic profiles of head and neck squamous cell carcinoma as predictable variables of tumor progression.
Massimo Rittà, Marco De Andrea, +8 authors, Santo Landolfo.
Head Neck, 2008 Dec 17; 31(3). PMID: 19072995
Information-theoretic analysis of phenotype changes in early stages of carcinogenesis.
F Remacle, Nataly Kravchenko-Balasha, Alexander Levitzki, R D Levine.
Proc Natl Acad Sci U S A, 2010 May 19; 107(22). PMID: 20479229    Free PMC article.
Prognostic Significance of Stromal Versus Intratumoral Infiltrating Lymphocytes in Different Subtypes of Breast Cancer Treated With Cytotoxic Neoadjuvant Chemotherapy.
Thaer Khoury, Vidya Nagrale, +3 authors, Song Yao.
Appl Immunohistochem Mol Morphol, 2017 Feb 12; 26(8). PMID: 28187033    Free PMC article.
Blockade of PD-1 Signaling Enhances Th2 Cell Responses and Aggravates Liver Immunopathology in Mice with Schistosomiasis japonica.
Sha Zhou, Xin Jin, +8 authors, Chuan Su.
PLoS Negl Trop Dis, 2016 Oct 30; 10(10). PMID: 27792733    Free PMC article.
pcaMethods--a bioconductor package providing PCA methods for incomplete data.
Wolfram Stacklies, Henning Redestig, +2 authors, Joachim Selbig.
Bioinformatics, 2007 Mar 09; 23(9). PMID: 17344241
Highly Cited.
Activation of OX40 augments Th17 cytokine expression and antigen-specific uveitis.
Zili Zhang, Wenwei Zhong, +6 authors, James T Rosenbaum.
Am J Pathol, 2010 Oct 19; 177(6). PMID: 20952591    Free PMC article.