associated head and neck squamous cell carcinomacancerres.aacrjournals.org/content/canres/early/2013/01/01/0008-5472... · 1 Evidence for a role of the PD-1:PD-L1 pathway in immune
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
1
Evidence for a role of the PD-1:PD-L1 pathway in immune resistance of HPV-
associated head and neck squamous cell carcinoma
Sofia Lyford-Pike1, Shiwen Peng2, Geoffrey D. Young1,3, Janis M. Taube2,4, William H. Westra1,2, Belinda Akpeng1, Tullia C. Bruno5, Jeremy D. Richmon1, Hao Wang6, Justin A. Bishop2, Lieping Chen7, Charles G. Drake5, 8, Suzanne L.Topalian3, 5, Drew M. Pardoll5, 9, Sara I. Pai1, 5, 10
Departments of 1Otolaryngology-Head and Neck Surgery, 2Pathology, 3Surgery, 4Dermatology, 5Oncology, 6Biostatistics, Johns Hopkins University School of Medicine and Sidney Kimmel Comprehensive Cancer Center, Baltimore, MD, USA; 7Department of Immunobiology, Yale University School of Medicine, New Haven, CT, USA; 8Department of Urology, James Buchanan Brady Urological Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA; and 9Immunology and Hematopoiesis Division, Sidney Kimmel Comprehensive Cancer Center, Baltimore, MD, USA
Running Title: PD-1:PD-L1 pathway in head and neck cancers
Keywords: PD-1; PD-L1; immune checkpoint; adapative resistance; HPV; head and neck cancers; oropharyngeal cancers; squamous cell carcinomas
Financial Support: This work was supported by the NIH/NIDCR P50 DE019032 Head and Neck Cancer SPORE grant (WHW, SIP), NIH/NIDCD T32000027 grant (SLP), and NIH R01 CA142779 (SLT).
10Corresponding author and to whom proofs and reprint requests should be sent: Sara I. Pai, MD, PhD 601 N. Caroline Street, JHOC 6th floor Baltimore, MD 21287 Phone: 410-502-9825 Fax: 410-955-0035 Email: [email protected]
Disclosures: Drs. Charles Drake and Suzanne Topalian have served as consultants to Bristol-Myers Squibb (BMS). Dr. Drake has licensed patents to BMS. Dr. Topalian has received research grants from BMS. Word Count: 4,614 Figures: 6; Supplemental Files: 4 Tables: 1
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
REFERENCES 1. Gillison ML, Koch WM, Capone RB, Spafford M, Westra WH, Wu L, et al. Evidence for a causal association between human papillomavirus and a subset of head and neck cancers. J Natl Cancer Inst 2000;92(9):709-20. 2. Chaturvedi Ak, Engels EA, Pfeiffer RM, Hernandez BY, Xiao W, Kim E, et al. Human papillomavirus and rising oropharyngeal cancer incidence in the United States. J Clin Oncol 2011;10;29(32):4294-301. 3. Begum S, Cao D, Gillison M, Zahurak M, Westra WH. Tissue distribution of human papillomavirus 16 DNA integration in patients with tonsillar carcinoma. Clin Cancer Res 2005;11(16):694-9. 4. Singhi AD and Westra WH. Comparison of human papillomavirus in situ hybridization and p16 immunohistochemistry in the detection of human papillomavirus-associated head and neck acner based on a prospective clinical experience. Cancer 2010;116(9):2166-73. 5. Gillison ML, D’Souza G, Westra W, Sugar E, Xiao W, Begum S, et al. Distinct risk factor profiles for human papillomavirus type 16-positive and human papilomavirus type 16-negative head and neck cancers. J Natl Cancer Inst 2008; 100(6):407-20. 6. Ang KK, Harris J, Wheeler R, Weber R, Rosenthal DI, Nguyen-Tan PF, et al. Human papillomavirus and survival of patients with oropharyngeal cancer. N Engl J Med 2010;363(1):24-35. 7. Westra WH. The changing face of head and neck cancer in the 21st century: the impact of HPV on the epidemiology and pathology of oral cancer. Head Neck Pathol. 2009;1:78-81. 8. Hodi FS, O’Day, SJ, McDermott DF, Weber RW, Sosman JA, Haanen JB, et al. Improved survival with ipilumimab in patients with metastatic melanoma. N Engl J Med 2010;363:711-23. 9. Brahmer JR, Drake CG, Wollner I, Powderly JD, Picus J, Sharfman WH, et al. Phase I study of single-agent anti-programmed death-1 (MDX-1106) in refractory solid tumors: safety, clinical activity, pharmacodynamics, and immunologic correlates. J Clin Oncol 2010;28:2167-75. 10. Topalian SL, Hodi FS, Brahmer JR, Gettinger SN, Smith DC, McDermott DF, et al. Safety, activity, and immune correlates of anti-PD-1 antibody in cancer. N Eng J Med 2012;366(26):2443-54.
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
11. Brahmer JR, Tykodi SS, Chow LQM, Hwu WJ, Topalian SL, Hwu P, et al. Safety and activity of anti-PD-L1 antibody in patients with advanced cancer. N Eng J Med 2012;366(26):2455-65. 12. Taube JM, Anders RA, Young GD, Xu H, Sharma R, McMiller TL, et al. Colocalization of inflammatory response with B7-H1 expression in human melanocytic lesions supports an adaptive resistance mechanism of immune escape. Sci Transl Med 2012 4(127):127-37. 13. Dong H, Strome SE, Salomao DR, Tamura H, Hirano F, Flies DB, et al. Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism of immune evasion. Nat Med 2002;8:793-800. 14. Strome SE, Dong H, Tamura H, Voxx, SG, Flies DB, Tamada K, et al. B7-H1 blockade augments adoptive T-cell immunotherapy for squamous cell carcinoma. Cancer Res 2003;63:6501-05. 15. Peng S, Lyford-Pike S, Akpeng B, Wu A, Hung CF, Hannaman D, et al. Low-dose cyclophosphamide administered as daily or single dose enhances the antitumor effects of a therapeutic HPV vaccine. Cancer Immunol Immunother 2012 Aug 4 [Epub ahead of print]. 16. Chen J, Li G, Meng H, Fan Y, Song Y, Wang S, et al. Upregulation of B7-H1 expression is associated with macrophage infiltration in hepatocellular carcinomas. Cancer Immunol Immunother 2012 61(1):101-8. 17. Zhao Q, Xiao X, Wu Y, Wei Y, Zhu LY, Zhou J, et al. Interleukin-17-educated monocytes suppress cytotoxic T cell function through B7-H1 in hepatocellular carcinoma patients. Eur J Immunol 2011;41:2314-22. 18. Parsa AT, Waldron JS, Panner A, Crane CA, Parney IF, Barry JJ, et al. Loss of tumor suppressor PTEN function increases B7-H1 expression and immunoresistance in glioma. Nat Med 2007;13(1):84-8. 19. Won HS, Jung CK, Chun SH, Kang JH, Kim YS, Sun DI, et al. Difference in expression of EGFR, pAkt, and PTEN between oropharyngeal and oral cavity squamous cell carcinoma. Oral Oncol 2012;48(1):985-990. 20. Agrawal N, Frederick MJ, Pickering CR, Gettegowda C, Chang K, Li RJ, et al. Exome sequencing of head and neck squamous cell carcinoma reveals inactivating mutations in NOTCH1. Science 2011;333(6046):1154-7. 21. Stransky N, Egloff AM, Tward AD, Kostic AD, Cibulskis K, Sivachenko A, et al. Science 2011;333(6046):1157-60.
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
22. Eerola Ak, Soini Y, Paakko P. A high number of tumor infiltrating lymphocytes are associated with a small tumor size, low tumor stage, and a favorable prognosis in operated small cell lung carcinoma. Clin Can Res 2000;6:1875-81. 23. Schumaher K, Haensch W, Roefzaad C, Schlag PM. Prognostic significance of activated CD8(+) T cell infiltrations within esophageal carcinomas. Cancer Res 2001;61:3932-6. 24. Zhang L, Conejo-Garcia JR, Katsarors D, Gimotty PA, Massobrio M, Regnani G, et al. Intratumoral T cells, recurrence, and survival in epithelial ovarian cancer. N Engl J Med 2003;348:203-13. 25. Galon J, Costes A, Sanchez-Cabo F, Kirilovsky A, Mlecnik B, Lagorce-Pages C, et al. Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science 2006;313:1960-4. 26. Hirano F, Kaneko K, Tamura H, Dong H, Wang S, Ichikawa M, et al. Blockade of B7-H1 and PD-1 by monoclonal antibodies potentiates cancer therapeutic immunity. Cancer Res 2005; 65(3):1089-96. 27. Porichis F, Kwon DS, Zupkosky J, Tighe DP, McMullen A, Brockman MA, et al. Responsiveness of HIV-specific CD4 T cells to PD-1 blockade. Blood 2011;118(4):965-74.
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
FIGURE LEGENDS Figure 1. The reticulated epithelium of benign tonsil tissue expresses high levels of PD-L1. A. Chronically inflamed tonsil tissue demonstrates localized PD-L1 expression within the reticulated epithelium of tonsillar crypts (long arrow). Magnification, x40. The inset (magnification, x400) demonstrates cell surface staining of the crypt epithelial cells. In contrast, the surface epithelium of the tonsils was negative for PD-L1 expression (short arrows) (B, C). Magnification, x40. Figure 2. High levels of PD-1 receptor on TILs from HPV-HNSCC. A. Representative flow cytometry of CD4+ T cell population expressing PD-1 in various tissues. Summary graph with mean frequency of CD4+PD-1(+) T cells in non-cancerous tonsils and HPV-HNSCC as compared to peripheral blood. B. Similar analysis performed for CD8+PD-1(+) T cell population. The circle on the representative flow data highlights a subpopulation of CD8+ TILs with high levels of PD-1 expression which was not observed in benign tonsils. (•) denotes chronic tonsillitis specimens and (�) denotes HPV-HNSCC. Figure 3. High levels of PD-L1 expression present in the tumor microenvironment of HPV-HNSCC. A. Hematoxylin and eosin stain of HPV-HNSCC demonstrates the proto-typic tumor nests (depicted by thin arrow) surrounded by a dense inflammatory stroma (depicted by thick arrow). Serial sections evaluated for B, HPV ISH (arrows mark areas of blue, intranuclear staining); C, p16 IHC; and D-F, PD-L1 IHC. Two patterns of PD-L1 staining were observed: peripheral tumoral staining (D, E) and diffuse intratumoral staining (F). Magnification, 400x Figure 4. Co-localization of TILs with PD-L1 expression in HPV-HNSCC. A. Hematoxylin and eosin stain of HPV-HNSCC (thin arrow marks tumor nests and thick arrow marks inflammatory stroma). Serial sections evaluated for: B, CD3; C, CD4; D, CD8; E, PD-L1; and F, CD68 expression. Red arrows indicate a representative area with clusters of PD-L1 and CD68 expression in serial sections (E, F). Magnification, 400x. Figure 5: Increased expression of IFN-� and CD8 mRNA in PD-L1(+) vs. (-) tonsil cancers. IFN-�, CD8, CD4, and GAPDH were evaluated by quantitative RT-PCR in both PD-L1(+) (n=3) and PD-L1(-) (n=6) tumors . Error bars represent standard error of the mean. Cycle threshold is on a log2 scale; lower numbers indicate greater expression. *p=0.003. **p=0.002. Results are representative of two separate experiments. Figure 6. PD-1 expressing CD8+ TILs are functionally anergic relative to peripheral blood PD-1 expressing T cells A. Representative flow cytometry comparing IFN-γ production by CD8+ T cells sorted by PD-1expression in PBMCs and TILs and stimulated with PMA/ionomycin. B. Summary graph of the ratio of IFN- γ production by PD-1(+) to PD-1(-) TILs and peripheral blood, in response to PMA/ionomycin. A value greater than 1 indicates increased IFN-γ production in PD-1(+) compared to PD-1(-) T cells, and a value less than 1 indicated a reduction. (•) denotes PBMC and (�) denotes TILs.
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384
Published OnlineFirst January 3, 2013.Cancer Res Sofia Lyford-Pike, Shiwen Peng, Geoffrey D Young, et al. carcinomaresistance of HPV-associated head and neck squamous cell Evidence for a role of the PD-1:PD-L1 pathway in immune
Updated version
10.1158/0008-5472.CAN-12-2384doi:
Access the most recent version of this article at:
To order reprints of this article or to subscribe to the journal, contact the AACR Publications
Permissions
Rightslink site. Click on "Request Permissions" which will take you to the Copyright Clearance Center's (CCC)
.http://cancerres.aacrjournals.org/content/early/2013/01/01/0008-5472.CAN-12-2384To request permission to re-use all or part of this article, use this link
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. Author Manuscript Published OnlineFirst on January 3, 2013; DOI: 10.1158/0008-5472.CAN-12-2384