YWHAG overexpression correlates with an immune-desert microenvironment and poor prognosis in laryngeal squamous cell carcinoma

Abstract

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.