Single-cell transcriptome profiling of the stepwise progression of head and neck cancer

Head and neck squamous cell carcinoma (HNSCC) undergoes stepwise progression from normal tissue to metastasized tumors. To comprehensively delineate the heterogeneity of tumor cells and their interactions during the progression of HNSCC, we employed single-cell RNA-seq (scRNA-seq) profiling for the tissues of normal, leukoplakia, HNSCC, and metastasized HNSCC. We identified novel clones of malignant cells expressing LGALS7B and fibroblasts expressing CXCL8, and the abundance of these cells in the tumor tissue was associated with poorer prognostic outcomes. In addition, we demonstrated that fibroblasts in leukoplakia and tumor tissue express COL1A1, which interacts with CD44+ malignant cells, facilitating HNSCC progression. CD4+FOXP3+ regulatory T cells expand in leukoplakia and express LAIR2 with collagen stimulation, which may provide a favorable microenvironment for tumor progression. In conclusion, stromal and immune cell interactions play critical roles in the stepwise progression of HNSCC, and we have identified steps during HNSCC progression for possible therapeutic interventions Overall design: Single-cell transcriptome profiling from 37 tissue specimens of non-tumoral surrounding normal tissue (NL, n = 9), leukoplakia (LP, n = 4), primary cancer (CA, n = 20), and metastatic tumors in lymph nodes (LN, n = 4)

头颈部鳞状细胞癌（Head and neck squamous cell carcinoma, HNSCC）会经历从正常组织到转移性肿瘤的逐步进展过程。为全面解析HNSCC进展过程中肿瘤细胞的异质性及其相互作用，我们对正常组织、白斑、HNSCC以及转移性HNSCC的组织样本开展了单细胞RNA测序（single-cell RNA-seq, scRNA-seq）分析。我们鉴定出了表达LGALS7B的恶性细胞新克隆，以及表达CXCL8的成纤维细胞新亚群；这类细胞在肿瘤组织中的丰度与较差的预后结局显著相关。此外，我们证实白斑组织与肿瘤组织中的成纤维细胞可表达COL1A1，该蛋白可与CD44+恶性细胞发生相互作用，进而促进HNSCC的进展。CD4+FOXP3+调节性T细胞会在白斑组织中扩增，并在胶原刺激下表达LAIR2，这可能为肿瘤进展营造了有利的微环境。综上，基质细胞与免疫细胞的相互作用在HNSCC的逐步进展中发挥关键调控作用，我们还鉴定出了HNSCC进展过程中可作为潜在治疗干预靶点的阶段。整体实验设计：本研究对37份组织标本开展了单细胞转录组分析，涵盖非肿瘤性癌旁正常组织（NL，n=9）、白斑（LP，n=4）、原发性癌（CA，n=20）以及淋巴结转移性肿瘤（LN，n=4）