YWHAG过表达与喉鳞状细胞癌免疫荒漠微环境及不良预后相关

文献信息

中文摘要

背景： 喉鳞状细胞癌（LSCC）仍是头颈部恶性肿瘤中癌症相关死亡的主要原因。肿瘤免疫微环境在决定疾病结局中起关键作用；然而，LSCC中特定的免疫细胞组成及其预后相关性尚未完全阐明。
方法： 应用CIBERSORT对来自381例TCGA-HNSCC样本的批量RNA测序数据进行分析，以估计22种免疫细胞类型的相对比例。选择LSCC（n = 64）进行深入分析。基于免疫谱的共识聚类将患者分为三个簇。在簇1（免疫荒漠）与簇2和3（免疫活跃）之间进行差异表达基因（DEGs）和通路富集分析。使用Kaplan-Meier生存分析进行预后评估。在一个独立的机构LSCC患者队列（n = 23）中，通过免疫组织化学评估YWHAG蛋白表达，并检查其与临床病理参数的相关性。
结果： 在HNSCC各亚部位中，LSCC肿瘤显示出显著高于其他解剖部位肿瘤的初始B细胞和浆细胞浸润（Kruskal-Wallis检验，校正P < 0.05）。LSCC的共识聚类识别出三种不同的免疫谱：簇1（免疫荒漠，以M0巨噬细胞和嗜酸性粒细胞为主）、簇2（免疫热，富集B细胞、浆细胞和CD8+ T细胞）和簇3（混合炎症型）。三级淋巴结构（TLS）相关基因在簇1中一致且显著下调（P < 0.05）。此外，通路分析显示B细胞相关通路在簇1中显著受到抑制。在I-III期肿瘤中的DEG分析显示YWHAG是簇1中唯一显著上调的基因。高YWHAG表达与所有分期中显著较短的总生存期相关（log-rank P < 0.05）。在机构免疫组织化学队列中，强YWHAG染色与较大的肿瘤大小显著相关。
结论： LSCC具有不同的免疫微环境亚型。富集B细胞和TLS相关基因特征的肿瘤表现出有利的结局，而YWHAG表达升高的免疫荒漠肿瘤与不良预后相关。YWHAG表达可能与不良结局相关，可能是由于抗肿瘤免疫反应减少和肿瘤大小增加。

英文摘要

BACKGROUND: Laryngeal squamous cell carcinoma (LSCC) remains a major cause of cancer-related mortality among head and neck malignancies. The tumor immune microenvironment plays a critical role in determining disease outcome; however, the specific immune cell composition and its prognostic relevance in LSCC have not been fully elucidated.
METHODS: CIBERSORT was applied to bulk RNA-sequencing data from 381 TCGA-HNSCC samples to estimate the relative fractions of 22 immune cell types. LSCCs (n = 64) were selected for in-depth analyses. Consensus clustering based on immune profiles stratified patients into three clusters. Differentially expressed genes (DEGs) and pathway enrichment analyses were performed between Cluster 1 (immune-desert) and Clusters 2 & 3 (immune-active). Kaplan-Meier survival analysis was used for prognostic assessment. YWHAG protein expression was evaluated by immunohistochemistry in an independent institutional cohort of patients with LSCC (n = 23), and its correlation with clinicopathological parameters was examined.
RESULTS: Across HNSCC subsites, LSCC tumors exhibited significantly higher infiltration of naive B cells and plasma cells than tumors from other anatomical sites (Kruskal-Wallis test, adjusted P < 0.05). Consensus clustering of LSCC identified three distinct immune profiles: Cluster 1 (immune-desert, dominated by M0 macrophages and eosinophils), Cluster 2 (immune-hot, enriched for B cells, plasma cells, and CD8+ T cells), and Cluster 3 (mixed-inflammatory). Tertiary lymphoid structure (TLS)-related genes were uniformly and significantly downregulated in Cluster 1 (P < 0.05). Furthermore, pathway analysis revealed that B cell-related pathways were significantly suppressed in Cluster 1. DEG analysis within Stage I-III tumors revealed YWHAG as the sole gene significantly upregulated in Cluster 1. High YWHAG expression was associated with significantly shorter overall survival across all stages (log-rank P < 0.05). In the institutional immunohistochemistry cohort, strong YWHAG staining was significantly associated with larger tumor size.
CONCLUSIONS: LSCC harbors distinct immune microenvironmental subtypes. Tumors enriched for B cell- and TLS- related gene signatures demonstrate favorable outcomes, whereas immune-desert tumors with elevated YWHAG expression are associated with poor prognosis. YWHAG expression may be associated with unfavorable outcomes, possibly due to reduced antitumor immune responses and increased tumor size.