Abstract 4750: The immunosuppressive tumor microenvironment (TME) in nasopharyngeal carcinoma: implications for immunotherapy
Authors: Duffield Amy S. , Ascierto Maria Libera , Anders Robert A. , Taube Janis M. , McMiller Tracee L. , Engle Elizabeth L. , Meeker Alan K. , Berger Alan E. , Pardoll Drew M. , Ambinder Richard F. , Topalian Suzanne L. 
Published: August 2018

Abstract
Nasopharyngeal carcinoma (NPC) is an EBV-driven tumor that shows variable expression of PD-L1 and ~20% objective response rate to anti-PD-1 monotherapy. As novel immune checkpoint inhibitors are being developed, combination therapies may allow for more effective treatment of both newly diagnosed and relapsed NPC. We characterized the TME in 13 cases of EBV+ NPC from the Johns Hopkins Pathology archives (7 primary tumors, 6 metastases). EBV status was confirmed with EBER ISH. Immunohistochemistry (IHC) was conducted on all cases for CD3, CD4, CD8, CD20, FoxP3, PD-1, PD-L1, LAG-3, TIM-3, GITR, IDO, COX2, and pSTAT3. Gene expression profiling (GEP) was performed on 7 cases with sufficient material (4 primary lesions, 3 metastases), using multiplex qRT-PCR for a panel of 61 candidate immune-related genes (Duffield, Blood Advances 2017). The immunosuppressive ligand PD-L1 was expressed on tumor cells in 11/13 cases (mean 22% tumor cells+, range 0-57%), as well as on infiltrating macrophages. The NPC inflammatory infiltrate was diverse, including CD4+, CD8+, CD20+ and CD68+ cells, and showed variable expression of immune-regulatory molecules. In all 13 cases, lymphocytes expressing PD-1 (mean 36% positive, range 8-70%), LAG-3 (7%; 1-30%) and GITR (12%; 2-27%) were found. FoxP3+ and TIM-3+ lymphocytes were infrequent. IDO+ macrophages were also infrequent; however, 7/13 NPCs showed expression of the immunosuppressive metabolic enzyme IDO by a proportion of tumor cells. Compared to 12 EBV+ Hodgkin lymphomas (Duffield, Blood Advances 2017), EBV+ NPCs demonstrated a Th17 cytokine profile with overexpression of IL1A, IL17RC, IL23A, and IL23R. The generation of pathogenic Th17 responses requires phosphorylation of the STAT3 transcription factor, and IHC confirmed that a subset of inflammatory cells in all NPC cases expressed pSTAT3 (mean 10%; range 1-40%). Additionally, upregulated gene expression characterizing activated macrophages was found (IDO, IL1A, IL12A, LYZ, TLR3). Of note, several molecules upregulated in the NPC TME are capable of inducing PD-L1 expression on human monocytes in vitro, including IL-1A and IL-32-gamma (Taube, Clin Cancer Res 2015; Duffield, Blood Advances 2017). Importantly PTGS2 (COX2), with known proinflammatory and immunosuppressive properties, was over-expressed in NPCs along with the downstream modulator CXCL8 (IL-8); IHC revealed COX2 expression in tumor cells but not infiltrating immune cells. In summary, NPC is characterized by markers of an immunosuppressive TME, including immune checkpoints and metabolic modulators. While these findings should be explored in a larger cohort, they have potential implications for designing combination NPC treatment regimens with anti-PD-1, which might include inhibitors of LAG-3, IDO, IL-17/-23, COX2, and/or IL-8. Funded by the Bristol-Myers Squibb International Immuno-Oncology Network and NCI R01 CA142779
Citation Format: Amy S. Duffield, Maria Libera Ascierto, Robert A. Anders, Janis M. Taube, Tracee L. McMiller, Elizabeth L. Engle, Alan K. Meeker, Alan E. Berger, Drew M. Pardoll, Richard F. Ambinder, Suzanne L. Topalian. The immunosuppressive tumor microenvironment (TME) in nasopharyngeal carcinoma: implications for immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4750.