Table_2_A 14-Marker Multiplexed Imaging Panel for Prognostic Biomarkers and Tumor Heterogeneity in Head and Neck Squamous Cell Carcinoma.docx

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.

头颈部鳞状细胞癌（head and neck squamous cell carcinoma, HNSCC）治疗领域的最新进展，凸显了开发新型预测工具以指导治疗策略的迫切需求。本研究旨在构建一种针对头颈部鳞状细胞癌的多重免疫组化（multiplex immunohistochemical, IHC）检测组合，用于评估该肿瘤的预后因素与瘤内异质性。为筛选可用于搭建新型多重免疫组化组合的免疫组化组织生物标志物，研究团队开展了系统综述与荟萃分析，对2008至2018年间报道的喉及咽部鳞状细胞癌免疫组化生物标志物进行梳理分析。本研究借助癌症基因组图谱（The Cancer Genome Atlas, TCGA）与Reactome通路数据库，对筛选获得的生物标志物的预后及功能意义进行验证。研究人员采用包含上述筛选标志物的14标志物显色型多重免疫组化组合，对未经治疗的头颈部鳞状细胞癌组织展开分析。最终纳入45项高质量研究，筛选得到31个候选组织生物标志物，总样本量N=7062。在TCGA喉及咽部鳞状细胞癌队列（N=205）中开展的预后验证结果显示，β-连环蛋白（β-catenin）、DKK1、PINCH1、ADAM10及TIMP1均与不良预后显著相关，上述标志物涉及免疫系统、细胞应答、细胞周期及发育系统等功能类别。研究团队将筛选得到的生物标志物整合构建14标志物检测组合，用于评估肿瘤生物标志物在组织结构中的异质性与极化表达模式，该组合与Wnt/β-连环蛋白通路的激活尤为相关。基于系统综述与荟萃分析的整合免疫组化分析，为评估头颈部鳞状细胞癌的侵袭性及瘤内异质性提供了一种原位蛋白质组学工具。