AKT3 Is a Novel Regulator of Cancer-Associated Fibroblasts in Head and Neck Squamous Cell Carcinoma

SIMPLE SUMMARY: Cancer-associated fibroblasts (CAFs) promote epithelial-to-mesenchymal transition, angiogenesis, and immunosuppression, resulting in tumor progression. The PI3K-AKT pathway is known to play vital roles in various cellular activities, including proliferation, growth, metabolism, and survival. In the present study, we sought to identify the key regulator of CAFs in head and neck squamous cell carcinoma (HNSCC) and elucidated the vital roles of AKT3, one of the AKT isoforms, in CAFs. A loss-of-function approach revealed that AKT3 in CAFs promoted their myofibroblastic phenotype and immunosuppressive characteristics. Moreover, the infiltration of AKT3-positive CAFs into tumors was positively correlated with that of various immune cells and an unfavorable prognosis in HNSCC patients. Our findings suggest that AKT3 is a potential biomarker to evaluate the CAF activity and immunosuppressive microenvironment in HNSCC. Furthermore, AKT3 is a potential target for cancer therapy that inhibits the pro-tumoral function of CAFs. ABSTRACT: Cancer-associated fibroblasts (CAFs) play vital roles in tumor progression by promoting epithelial-to-mesenchymal transition, angiogenesis, and immunosuppression. In the present study, we sought to identify the key regulators of the pro-tumoral functions of CAFs in head and neck squamous cell carcinoma (HNSCC). mRNA expression data obtained from The Cancer Genome Atlas revealed that CAF-specific mRNA expression correlated with genes that relate to an immunosuppressive microenvironment in a HNSCC cohort. RNA sequencing of CAFs and normal fibroblasts isolated from HNSCC specimens identified 1127 differentially expressed genes (DEGs) and several upregulated pathways in CAFs. Among the 1127 DEGs, we identified 13 immune function-related genes and focused on AKT3 as a potential regulator of CAFs. The targeted depletion of AKT3 in CAFs revealed that AKT3 promotes their myofibroblastic phenotype. AKT3-transduced CAFs exhibited downregulated the expression of immunosuppressive cytokine genes, impairing T-cell suppression and pro-tumoral macrophage induction. The immunohistochemistry of 72 HNSCC patients showed that AKT3 expression in CAFs positively correlated with tumor infiltration by CAFs, tumor-associated macrophages, dendritic cells, and T cells. Moreover, AKT3 expression in CAFs was an independent prognostic factor for overall survival. In conclusion, AKT3 is a potential target for cancer therapy that inhibits the pro-tumoral function of CAFs and reverses CAF-mediated immunosuppression.