Recurrent HNSCC harbor an immunosuppressive tumor immune microenvironment suggesting successful tumor immune evasion.

PURPOSE Recurrent tumors (RTs) of head and neck squamous cell carcinoma (HNSCC) occur in up to 60% with poor therapeutic response and detrimental prognosis. We hypothesized that HNSCC RTs successfully evade anti-tumor immune response and aimed to reveal tumor immune microenvironment (TIME) changes of primary tumors (PTs) and corresponding RTs. EXPERIMENTAL DESIGN Tumor infiltrating leukocytes (TILs) of 300 PTs and 108 RTs from two large, independent and clinically well-characterized HNSCC cohorts (discovery cohort/DC, validation cohort/VD) were compared by immunohistochemistry. mRNA expression analysis of 730 immune-related genes was performed for 18 PTs and RTs after adjuvant chemoradiotherapy (CRT). The effect of chemotherapy- and radiation-resistance was assessed with an in vitro spheroid/immunocyte coculture model. RESULTS TIME analysis revealed overall decrease of TILs with significant loss of CD8+ T-cells (DC p=0.045/VC p<0.0001) and B-lymphocytes (DC p=0.036/VC p<0.0001) in RTs compared to PTs in both cohorts. Decrease predominantly occurred in RTs after CRT. Gene expression analysis confirmed loss of TILs (p=0.0004) and B-lymphocytes (p<0.0001) and showed relative increase of neutrophils (p=0.018), macrophages (p<0.0001), dendritic cells (p=0.0002) and mast cells (p=0.0057) as well as lower overall expression of immune related genes (p=0.018) in RTs after CRT. Genes involved in B-lymphocyte functions and number of tertiary lymphoid structures showed the strongest decrease. SPP1 and MAPK1 were upregulated in vivo and in vitro indicating their potential suitability as therapeutic targets in CRT resistance. CONCLUSIONS HNSCC RTs have an immunosuppressive TIME, which is particularly apparent after adjuvant CRT and might substantially contribute to poor therapeutic response and prognosis.