Investigating cancer-associated fibroblast and NK cell interactions in head and neck cancer

Abstract

Head and neck cancer is an aggressive malignant neoplasm, usually diagnosed at advanced stage and associated with a poor prognosis. Recently, immunotherapy has shown promising results in controlling the disease outcome and overall survival of these patients, but the clinical response is still limited. In this context, the tumor microenvironment (TME), especially the cancer-associated fibroblasts (CAFs), plays a fundamental role in modulating the antitumor immune response, favoring tumor progression, evasion and resistance to immunotherapy. Among the anti-tumor effector cells, Natural Killer (NK) cells have an important role and many therapeutic strategies have been investigated to improve NK cell activation, cytotoxicity and tumor infiltration. However, how CAFs impairs NK cell function in the TME is poorly understood. Thus, the aim of this study is to understand the mechanisms involved in the interaction of CAFs with NK cells in head and neck cancer. In this way, data from single cell RNA sequencing (scRNAseq) from head and neck tumors will be used to characterize the phenotype and function of CAFs and NK cells as well as to establish interaction pathways and regulatory gene networks between both cell types. The spatial relationships of these cells will be evaluated by multiplexed immunohistochemistry in tissue sections of head and neck cancer. Additionally, cell lines derived from head and neck cancer, primary CAFs, the cell line NK92 and fresh NK isolated from healthy donors will be used to carry out in vitro assays, with a 2D, 3D and organotypic tumor model approaches as well as in vivo assays, in order to investigate the effects of CAFs on NK cell activation, cytotoxicity and infiltration. Finally, we will evaluate the NK anti-tumor response after treatment with Setanexib, a specific CAFs inhibitor, in a mouse model of head and neck cancer. (AU)