Soluble Immune Biomarkers as a Diagnostic Tool of Head and Neck Squamous Cell Carcinoma Histological Inflammatory Subtypes

Abstract

Head and neck squamous cell carcinoma (HNSCC) is a devastating malignancy that occurs in close proximity to vital structures. Despite all advances in diagnostic and therapeutic measures, the overall 5-year survival rate stays at ~65% and can be dismal for recurrent and advanced stages. There is accumulating evidence indicating the immune suppressive potential of HNSCC by which it can escape and/ or suppress the immune system. The recent advent of immunotherapy showed unprecedented improvement in overall response of advanced stage HNSCC. Yet, the overall response rate of HNSCC to immunotherapy remains at ~ 15%. Hence, further understanding of HNSCC tumor inflammation is warranted. The hypothesis of the current work is that the histological inflammatory subtype (HIS) of HNSCC can provide the basis for patient stratification that can be distinguished from the cytokine profile in peripheral blood, therefore better treatment strategies and patient outcomes. Paired tumor tissue and plasma of 104 HNSCC cases were collected according to UMB IRB protocol. Scoring of the HIS subtype was carried out using immunohistochemistry (IHC) of the emerging immune biomarker Semaphorin 4D (Sema4D). Our cohort showed 52% HIS inflamed (HIS-INF), 40% immune excluded (HIS-IE) and 8% deserted (HIS-ID). Differential gene expression (DGE) analysis of 10 HNSCC tumor tissues using NanoString immune-oncologic (human IO-360) 700 genes set showed that tumors with HIS-IE clustered as IFN-γ negative/ low immune signature compared to HIS-INF and were higher in hypoxia gene expression and the myeloid cellular compartment, while the HIS-INF demonstrated higher lymphoid component. DGE revealed a novel association between the high soluble Sema4D in plasma (HsS4D) and higher transcriptional level of Osteopontin (OPN) in the tumor tissue, that we also demonstrated in vitro. Furthermore, using ELISA-Luminex™ system, HIS-IE tumor tissue was significantly distinguished from the HIS-INF, in almost 40 traditional cytokines detected in the paired plasma samples. In conclusion, our work demonstrated a stratification model of HNSCC based on the underlying HIS profile that can be detected via the soluble cytokine panel in blood. These findings can open new avenues for patient stratification and enhance personalized clinical care in the field of HNSCC.