ATP/P2X7信号通路作为鼻息肉早期复发的潜在预测生物标志物：与炎症小体激活及上皮屏障功能障碍的相关性

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中文摘要

背景： 慢性鼻窦炎伴鼻息肉（CRSwNP）以显著的上皮重塑和屏障功能障碍为特征，导致术后复发率高。然而，与早期复发相关的分子机制仍知之甚少。
方法： 我们对同一患者初次手术前和复发时获得的鼻息肉组织进行了配对蛋白质组学分析（发现队列），以识别复发相关蛋白。在独立队列中验证候选蛋白，并通过受试者工作特征（ROC）曲线和Kaplan-Meier分析评估其与复发风险的关联。通过免疫荧光进一步表征所识别候选蛋白的表达和细胞定位。使用ATP刺激和特异性药理学抑制剂在原代人鼻上皮细胞中进行机制研究。
结果： 配对蛋白质组学分析确定P2X7、PDE4D和JADE2为复发组织中上调最高的前3种蛋白。其中，只有P2X7在验证队列中持续升高，并且主要定位于鼻上皮（P < 0.01）。ROC曲线分析表明，组织P2X7水平可能作为CRSwNP术后复发的潜在预测标志物（AUC = 0.678，P = 0.022）。Kaplan-Meier分析进一步显示，高P2X7表达组的复发风险显著更高（HR = 2.323，P = 0.015）。复发组织表现出NLRP3和IL-1β水平升高，上皮屏障标志物occludin和E-cadherin表达降低，并伴有细胞外ATP浓度增加。在体外，ATP刺激上调P2X7表达，诱导NLRP3和切割的IL-1β，并下调occludin和E-cadherin。值得注意的是，这些效应被P2X7特异性抑制剂（A-438079）逆转。
结论： 配对蛋白质组学分析揭示了复发前和复发后鼻息肉之间不同的蛋白表达谱，P2X7可作为早期复发的有前景的预测生物标志物。在机制上，ATP/P2X7轴可能通过促进炎症小体激活和上皮屏障破坏来驱动疾病复发。

英文摘要

BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by pronounced epithelial remodeling and barrier dysfunction, contributing to a high rate of postoperative recurrence. However, molecular mechanisms associated with early recurrence remain poorly understood.
METHODS: We performed paired proteomic profiling of nasal polyp tissues obtained from the same patients before initial surgery and at the time of recurrence (discovery cohort) to identify recurrence-associated proteins. Candidate proteins were validated in an independent cohort, and their association with recurrence risk was assessed by receiver operating characteristic (ROC) curve and Kaplan-Meier analyses. The expression and cellular localization of the identified candidate were further characterized by immunofluorescence. Mechanistic investigations were conducted in primary human nasal epithelial cells using ATP stimulation and specific pharmacological inhibitors.
RESULTS: Paired proteomic analysis identified P2X7, PDE4D, and JADE2 as the top 3 upregulated proteins in recurrent tissues. Among these, only P2X7 was consistently elevated in the validation cohort and predominantly localized to the nasal epithelium (P < 0.01). ROC curve analysis indicated that tissue P2X7 level might serve as a potential predictive marker for postoperative recurrence of CRSwNP. (AUC = 0.678, P = 0.022). Kaplan-Meier analysis further showed that the risk of recurrence was significantly higher in the high P2X7 expression group (HR = 2.323, P = 0.015). Recurrent tissues exhibited elevated levels of NLRP3 and IL-1β, reduced expression of epithelial barrier markers occludin and E-cadherin, accompanied by increased extracellular ATP concentrations. In vitro, ATP stimulation upregulated P2X7 expression, induced NLRP3 and cleaved IL-1β, and downregulated occludin and E-cadherin. Notably, these effects were reversed by a P2X7-specific inhibitor (A-438079).
CONCLUSION: Paired proteomic profiling revealed distinct protein expression profiles between pre- and post-recurrence nasal polyps, with P2X7 serving as a promising predictive biomarker for early recurrence. Mechanistically, the ATP/P2X7 axis may drive disease recurrence by potentially promoting inflammasome activation and epithelial barrier disruption.