Background/Objectives:Head and neck squamous cell carcinoma (HNSCC) is a biologically heterogeneous malignancy with poor outcomes in advanced disease. Increasing evidence indicates that the tumor microenvironment, particularly cancer-associated fibroblasts (CAFs), plays an important role in tumor progression and immune regulation. However, the diversity of CAF subsets and their clinical relevance in HNSCC remain incompletely understood. This study aimed to characterize CAF heterogeneity and assess the prognostic significance of CAF subset-specific transcriptional programs.Methods:Single-cell RNA sequencing data from HNSCC tumors were analyzed to identify CAF subsets based on differentially expressed genes. CAF subset-specific gene signatures were used to construct prognostic risk models for overall survival (OS) and progression-free survival (PFS) in The Cancer Genome Atlas HNSCC cohort, with validation in an independent dataset. CAF-driven prognostic groups were defined, and their immune landscapes and biological pathways were evaluated. Bulk RNA sequencing of primary CAF cultures was performed for validation.Results:Six CAF subsets were identified, including myofibroblastic (myCAF), inflammatory (iCAF), antigen-presenting, and extracellular matrix-related CAFs. Risk scores derived from inflammatory CAF subsets consistently predicted shorter OS across independent cohorts, whereas PFS prediction showed greater cohort dependency. CAF-based stratification identified patient subgroups with distinct immune profiles and pathway enrichment patterns. These results were supported by validation analyses and by bulk RNA sequencing of primary CAFs, demonstrating preservation of myCAF- and iCAF-like transcriptional programs ex vivo.Conclusions:CAF heterogeneity has important prognostic and immunological implications in HNSCC. Inflammatory CAF-related transcriptional programs represent robust markers of patient survival and may complement tumor-intrinsic biomarkers.
背景/目的：头颈鳞状细胞癌（HNSCC）是一种生物学异质性恶性肿瘤，晚期患者预后较差。越来越多的证据表明，肿瘤微环境，特别是癌症相关成纤维细胞（CAFs），在肿瘤进展和免疫调节中发挥重要作用。然而，CAFs亚群的多样性及其在HNSCC中的临床相关性仍未完全明确。本研究旨在描述CAFs的异质性，并评估CAFs亚群特异性转录程序的预后意义。
方法：通过分析HNSCC肿瘤的单细胞RNA测序数据，基于差异表达基因识别CAFs亚群。利用CAFs亚群特异性基因特征，在癌症基因组图谱（TCGA）HNSCC队列中构建总生存期（OS）和无进展生存期（PFS）的预后风险模型，并在独立数据集中进行验证。确定了CAFs驱动的预后分组，并评估了其免疫景观和生物学通路。通过对原代CAFs培养物进行批量RNA测序以验证结果。
结果：共识别出六种CAFs亚群，包括肌成纤维细胞性CAF（myCAF）、炎症性CAF（iCAF）、抗原呈递性CAF及细胞外基质相关CAF。源自炎症性CAF亚群的风险评分在不同独立队列中均一致预测较短的OS，而PFS预测则显示出更大的队列依赖性。基于CAFs的分层确定了具有不同免疫特征和通路富集模式的患者亚组。验证分析及原代CAFs的批量RNA测序结果支持了上述发现，证明myCAF和iCAF样转录程序在体外培养中得以保留。
结论：CAFs的异质性在HNSCC中具有重要的预后和免疫学意义。炎症性CAF相关的转录程序可作为患者生存的可靠标志物，并可能补充肿瘤内在生物标志物。
肿瘤(癌症)患者之家