多组学与机器学习鉴定ALOX5作为慢性鼻窦炎伴鼻息肉中铁死亡相关上皮屏障功能障碍驱动因子

文献信息

中文摘要

慢性鼻窦炎伴鼻息肉（CRSwNP）是上气道常见的慢性炎症性疾病，其潜在发病机制仍不完全清楚。本研究以内质网应激（ERS）相关基因集为分析起点，系统筛选并鉴定与CRSwNP密切相关的核心基因。通过整合加权基因共表达网络分析（WGCNA）、差异表达分析和多种机器学习算法，鉴定出4个枢纽基因（ALOX5、HMOX1、PXDN和CARD9），这些基因对CRSwNP均表现出良好的诊断性能（AUC = 0.863-0.962，P < 0.05）。采用RT-qPCR、Western blotting和免疫组织化学验证这些枢纽基因在CRSwNP组织中的异常表达。值得注意的是，ALOX5的表达水平与CRSwNP的临床严重程度呈显著正相关（R = 0.40-0.41，P < 0.01）。单细胞转录组分析进一步揭示，铁死亡相关特征在CRSwNP患者的鼻上皮细胞中显著富集。免疫荧光染色和原代鼻上皮细胞实验表明，ALOX5表达与上皮紧密连接蛋白呈负相关，且靶向ALOX5可有效调节铁死亡过程。总之，本研究鉴定了在CRSwNP中具有潜在诊断价值和功能相关性的枢纽基因，并阐明了ALOX5介导的上皮铁死亡可能参与疾病进展，从而为CRSwNP的精准诊断和靶向治疗提供了新见解。

英文摘要

Chronic rhinosinusitis with nasal polyps (CRSwNP) is a common chronic inflammatory disease of the upper airway, and its underlying pathogenesis remains incompletely understood. In this study, endoplasmic reticulum stress (ERS)-related gene sets were employed as the analytical starting point to systematically screen and identify core genes closely associated with CRSwNP. Through the integration of weighted gene co-expression network analysis (WGCNA), differential expression analysis, and multiple machine learning algorithms, 4 hub genes (ALOX5, HMOX1, PXDN, and CARD9) were identified, all of which exhibited favorable diagnostic performance for CRSwNP (AUC = 0.863-0.962, P < 0.05). RT-qPCR, Western blotting, and immunohistochemistry were performed to validate the aberrant expression of these hub genes in CRSwNP tissues. Notably, the expression level of ALOX5 was significantly positively correlated with the clinical severity of CRSwNP (R = 0.40-0.41, P < 0.01). Single-cell transcriptomic analysis further revealed that ferroptosis-related signatures were significantly enriched in nasal epithelial cells from patients with CRSwNP. Immunofluorescence staining and primary nasal epithelial cell experiments indicated that ALOX5 expression was negatively correlated with epithelial tight junction proteins and that targeting ALOX5 effectively modulated the ferroptosis process. Collectively, this study identified hub genes with potential diagnostic value and functional relevance in CRSwNP and elucidated the potential involvement of ALOX5-mediated epithelial ferroptosis in disease progression, thereby providing novel insights into the precise diagnosis and targeted therapy of CRSwNP.