A High-Throughput Immune-Oncology Screen Identifies EGFR Inhibitors as Potent Enhancers of Antigen-Specific Cytotoxic T-lymphocyte Tumor Cell Killing
Journal
Cancer Immunology Research
Journal Volume
6
Journal Issue
12
Pages
1511-1523
Date Issued
2018
Author(s)
Lizotte P.H.
Luster T.A.
Cavanaugh M.E.
Taus L.J.
Wang S.
Dhaneshwar A.
Mayman N.
Yang A.
Kulkarni M.
Badalucco L.
Fitzpatrick E.
Kuraguchi M.
Bittinger M.
Kirschmeier P.T.
Gray N.S.
Barbie D.A.
Janne P.A.
Abstract
We developed a screening assay in which luciferized ID8 expressing OVA was cocultured with transgenic CD8? T cells specifically recognizing the model antigen in an H-2b–restricted manner. The assay was screened with a small-molecule library to identify compounds that inhibit or enhance T cell–mediated killing of tumor cells. Erlotinib, an EGFR inhibitor, was the top compound that enhanced T-cell killing of tumor cells. Subsequent experiments with erlotinib and additional EGFR inhibitors validated the screen results. EGFR inhibitors increased both basal and IFNg-induced MHC class-I presentation, which enhanced recognition and lysis of tumor cell targets by CD8? cytotoxic T lymphocytes. The ID8 cell line was also transduced to constitutively express Cas9, and a pooled CRISPR screen, utilizing the same target tumor cell/T-cell assay, identified single-guide (sg)RNAs targeting EGFR that sensitized tumor cells to T cell–mediated killing. Combination of PD-1 blockade with EGFR inhibition showed significant synergistic efficacy in a syngeneic model, further validating EGFR inhibitors as immunomodulatory agents that enhance checkpoint blockade. This assay can be screened in high-throughput with small-molecule libraries and genome-wide CRISPR/Cas9 libraries to identify both compounds and target genes, respectively, that enhance or inhibit T-cell recognition and killing of tumor cells. Retrospective analyses of squamous-cell head and neck cancer (SCCHN) patients treated with the combination of afatinib and pembrolizumab demonstrated a rate of clinical activity exceeding that of each single agent. Prospective clinical trials evaluating the combination of an EGFR inhibitor and PD-1 blockade should be conducted. ?2018 American Association for Cancer Research.
SDGs
Other Subjects
afatinib; epidermal growth factor receptor kinase inhibitor; erlotinib; major histocompatibility antigen class 1; pembrolizumab; programmed death 1 receptor; afatinib; antineoplastic agent; epidermal growth factor receptor; firefly luciferase; monoclonal antibody; PDCD1 protein, human; pembrolizumab; programmed death 1 receptor; protein kinase inhibitor; animal cell; animal experiment; animal model; antigen specificity; Article; cancer patient; cancer research; cancer screening; CD8+ T lymphocyte; cell killing; clustered regularly interspaced short palindromic repeat; computer assisted tomography; controlled study; CRISPR-CAS9 system; cytolysis; cytotoxic T lymphocyte; drug efficacy; drug potentiation; enhancer region; enzyme linked immunosorbent assay; flow cytometry; genome-wide association study; head and neck cancer; high throughput screening; high throughput sequencing; hypopharynx squamous cell carcinoma; ID8 cell line; immunomodulation; in vivo study; larynx squamous cell carcinoma; medical record; molecular library; monotherapy; mouse; mouth squamous cell carcinoma; nonhuman; nuclear magnetic resonance imaging; oncology; open reading frame; oropharynx squamous cell carcinoma; retrospective study; target cell; treatment duration; tumor cell; animal; C57BL mouse; coculture; CRISPR Cas system; cytotoxic T lymphocyte; drug effect; drug screening; genetics; head and neck tumor; high throughput screening; human; immunology; procedures; tumor cell line; Afatinib; Animals; Antibodies, Monoclonal, Humanized; Antineoplastic Combined Chemotherapy Protocols; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Coculture Techniques; CRISPR-Cas Systems; Drug Screening Assays, Antitumor; ErbB Receptors; Head and Neck Neoplasms; High-Throughput Screening Assays; Humans; Luciferases, Firefly; Mice, Inbred C57BL; Programmed Cell Death 1 Receptor; Protein Kinase Inhibitors; Squamous Cell Carcinoma of Head and Neck; T-Lymphocytes, Cytotoxic
Publisher
American Association for Cancer Research Inc.
Type
journal article