Abstract 1748: Head and neck cancer-derived exosomes facilitate carcinogenesis in the murine 4NQO chemically induced oropharyngeal cancer
Authors: Razzo Beatrice M. , Hong Chang-Sook , Fabian Kellsye P. , Ludwig Nils , Sharma Priyanka , Storkus Walter J. , Whiteside Theresa L. 
Published: August 2018

Purpose: An increasing body of evidence suggests that tumor-derived exosomes (TEX) promote tumor formation and progression. This study aims to show that exosomes produced by the SCCVII murine tumor cell line promote progression of pre-malignant oropharyngeal lesions to malignant tumors. The 4-Nitroquinoline 1-oxide (4NQO) tumor model is chemically induced in immunocompetent mice. It recapitulates all the histological stages and molecular changes associated with of oral carcinogenesis described for human head and neck squamous cell carcinoma (HNSCC). Experimental design: Exosomes were isolated from supernatant of a murine as well as human HNSCC cell-lines by mini size-exclusion chromatography (miniSEC). Morphology, number, protein and the molecular cargo of the recovered exosomes were determined. Exosome-mediated in vitro inhibition of lymphocyte proliferation and induction of CD8+ T-cell apoptosis were demonstrated. C57BL/6 mice were treated with 4NQO supplied in drinking water (100ug/ml) for 16 weeks and all developed oral and esophageal pre-malignant lesions. Exosomes were administered via a single tail-vein or retro-orbital injection at week 18. Tumor sizes and overall tumor burden per mouse were measured with a caliper. Proliferation indexes of individual tumors were measured by immunohistochemistry. Splenocytes were harvested and were analyzed for the presence and frequency of myeloid derived suppressor cells (MDSC) and Tregs, and for in vitro responsiveness of T cells to exogenous activators. Results: SCCVII-derived exosomes were found to carry an immunosuppressive cargo, including CD39, CD73, PD-L1, FasL and COX-2. One-time 90ug injections (IV) of exosomes to groups of &gt;8 mice induced a statistically significant increase in the number of developing tumors and in the overall tumor burden per mouse, relative to control mice injected with PBS. The average number of tumors per mouse was 6.3 versus 3.6, respectively (p &lt; 0.005). TEX derived from human HNSCC cell lines also induced a greater though not statistically significant number of tumors in 4NQO-treated mice. The MDSC frequency was increased in the exosome-injected mice relative to controls. The Treg frequency was not altered. Conclusions: The data demonstrate that the delivery of Head and Neck Cancer-derived exosomes enhanced carcinogenesis of the 4NQO-induced tumors in mice. Augmented carcinogenesis was associated with systemic immunosuppression and increases in the frequency of MDSCs in spleens of mice treated with exosomes.

Citation Format: Beatrice M. Razzo, Chang-Sook Hong, Kellsye P. Fabian, Nils Ludwig, Priyanka Sharma, Walter J. Storkus, Theresa L. Whiteside. Head and neck cancer-derived exosomes facilitate carcinogenesis in the murine 4NQO chemically induced oropharyngeal cancer [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 1748.