A 14-Marker Multiplexed Imaging Panel for Prognostic Biomarkers and Tumor Heterogeneity in Head and Neck Squamous Cell Carcinoma

Abstract

Recent advances made in treatment for head and neck squamous cell carcinoma (HNSCC) highlight the need for new prediction tools to guide therapeutic strategies. In this study, we aimed to develop a HNSCC-targeting multiplex immunohistochemical (IHC) panel that can evaluate prognostic factors and the intratumor heterogeneity of HNSCC. To identify IHC-based tissue biomarkers that constitute new multiplex IHC panel, a systematic review and meta-analysis were performed to analyze reported IHC biomarkers in laryngeal and pharyngeal SCC in the period of 2008–2018. The Cancer Genome Atlas (TCGA) and Reactome pathway databases were used to validate the prognostic and functional significance of the identified biomarkers. A 14-marker chromogenic multiplex IHC panel including identified biomarkers was used to analyze untreated HNSCC tissue. Forty-five high-quality studies and thirty-one candidate tissue biomarkers were identified (N = 7062). Prognostic validation in TCGA laryngeal and pharyngeal SCC cohort (N = 205) showed that β-catenin, DKK1, PINCH1, ADAM10, and TIMP1 were significantly associated with poor prognosis, which were related to functional categories such as immune system, cellular response, cell cycle, and developmental systems. Selected biomarkers were assembled to build a 14-marker panel, evaluating heterogeneity and polarized expression of tumor biomarkers in the tissue structures, which was particularly related to activation of Wnt/β-catenin pathway. Integrated IHC analysis based on a systemic review and meta-analysis provides an in situ proteomics tool to assess the aggressiveness and intratumor heterogeneity of HNSCC.

Harnessing Oncolytic Extracellular Vesicles for Tumor Cell‐Preferential Cytoplasmic Delivery of Misfolded Proteins for Cancer Immunotherapy

“…To better understand the heterogeneous TME, we used multiplex immunohistochemistry of tumor tissues. [36] Consistent with the immune profile data acquired from flow cytometry, bRSVF-EV treatment notably incremented the population of CD8 + T cells, which spread throughout the tumor tissues (Figure 4I). Collectively, these findings demonstrate that bRSVF-EV can effectively inhibit tumor growth in vivo by activating an antitumor immune response.…”

The role of tetraspanins pan-cancer

“…From this, we propose a new hypothesis. ADAM10 has now been shown to exert a vital part in tumorigenesis and tumor development in HNSC ( Huang et al., 2014 ; Loganathan et al., 2020 ; Mitsuda et al., 2021 ). In our study, consistent with LIHC, TSPAN15 was relatively highly expressed in HNSC ( Figures 2 B and 2D), with poorer survival outcomes ( Figure 3 C) and high correlation with stromal and immune scores ( Figure 7 C), suggesting that TSPAN15 may also play a role in HNSC.…”

Molecular and nutritional markers in head and neck cancer

“…This strategy was recently applied by Mitsuda et al . [10 ▪▪ ] who built in SCCHN a 14-marker panel particularly related to the activation of Wnt/β-catenin pathway. Interestingly, the developed panel enabled a simultaneous measurement of tumor, immune and stroma biomarkers on a single tumor slide.…”

“…Notably, a single quantitative assessment was done using image cytometry. Also of interest, the approach developed by the authors permitted to identify multiple biomarkers-positive cancer cells, which might be related to the presence of aggressive subpopulations and localized at the invasive margin of studied tumors [10 ▪▪ ]. The next step beyond the establishment of such a multimarker panel would be to leverage the multiple expression profile through an appropriate biostatistical analysis allowing a global prognostic index to be established.…”