Abstract：
BACKGROUND: Cilia loss and impaired motile ciliary functions are among the typical pathological features of chronic rhinosinusitis with nasal polyps (CRSwNP). IL17A and IL22 are the canonical cytokines of type 3 inflammation, exhibiting similar functional effects on epithelial cells. In this study, we sought to examine the effects of IL17A and IL22 on ciliated cells and investigate the potential involvement of Hippo-YAP signaling in their influence on ciliogenesis.
METHODS: We assessed both the mRNA and protein expression levels of IL17A and IL22 in nasal tissues obtained from patients with CRSwNP and compared them to those from healthy controls. To further explore the impact of IL17A and IL22, we established a primary human nasal epithelial cell model using different concentrations (2 ng/mL, 10 ng/mL, 50 ng/mL) for a duration of 28 days in an air-liquid interface culture. Additionally, we employed the inhibitor verteporfin to investigate whether IL17A and IL22 exert their effects on ciliated cells via the Hippo-YAP pathway.
RESULTS: The mRNA and protein levels of IL17A and IL22 in CRSwNP were significantly higher than those in healthy controls, revealing a robust correlation between IL17A and IL22. YAP was highly expressed in the nucleus of ciliated cells in CRSwNP and displayed a positive correlation with clinical symptoms. Both IL17A and IL22 were found to reduce the number of ciliated cells. IL17A, but not IL22, suppressed ciliogenesis by disrupting the proper development and docking of the basal body of ciliated cells, resulting in motile ciliary dysfunctions. Furthermore, the expression of YAP within the nucleus of ciliated cells gradually declined as these cells reached the final stage of differentiation. However, this process was obstructed by IL17A only. YAP inhibitors, such as verteporfin, markedly reversed the effects of IL17A by increasing the proportion of ciliated cells, suppressing nuclear YAP expression in these cells, and enhancing ciliary beating frequency.
CONCLUSIONS: Both IL17A and IL22 are overexpressed in nasal epithelium of CRSwNP, which is associated with the impairment of epithelial cell differentiation. Furthermore, IL17A has been shown to exert a disruptive effect on morphogenesis of motile cilia via activation of YAP.
METHODS: We assessed both the mRNA and protein expression levels of IL17A and IL22 in nasal tissues obtained from patients with CRSwNP and compared them to those from healthy controls. To further explore the impact of IL17A and IL22, we established a primary human nasal epithelial cell model using different concentrations (2 ng/mL, 10 ng/mL, 50 ng/mL) for a duration of 28 days in an air-liquid interface culture. Additionally, we employed the inhibitor verteporfin to investigate whether IL17A and IL22 exert their effects on ciliated cells via the Hippo-YAP pathway.
RESULTS: The mRNA and protein levels of IL17A and IL22 in CRSwNP were significantly higher than those in healthy controls, revealing a robust correlation between IL17A and IL22. YAP was highly expressed in the nucleus of ciliated cells in CRSwNP and displayed a positive correlation with clinical symptoms. Both IL17A and IL22 were found to reduce the number of ciliated cells. IL17A, but not IL22, suppressed ciliogenesis by disrupting the proper development and docking of the basal body of ciliated cells, resulting in motile ciliary dysfunctions. Furthermore, the expression of YAP within the nucleus of ciliated cells gradually declined as these cells reached the final stage of differentiation. However, this process was obstructed by IL17A only. YAP inhibitors, such as verteporfin, markedly reversed the effects of IL17A by increasing the proportion of ciliated cells, suppressing nuclear YAP expression in these cells, and enhancing ciliary beating frequency.
CONCLUSIONS: Both IL17A and IL22 are overexpressed in nasal epithelium of CRSwNP, which is associated with the impairment of epithelial cell differentiation. Furthermore, IL17A has been shown to exert a disruptive effect on morphogenesis of motile cilia via activation of YAP.
摘要：
背景：纤毛丢失和纤毛运动功能受损是慢性鼻-鼻窦炎伴鼻息肉（CRSwNP）的典型病理特征之一。白细胞介素-17A(IL-17A)和白细胞介素-22(IL-22)是3型炎症的典型细胞因子,对上皮细胞表现出类似的功能作用。在这项研究中,我们试图检查IL-17A和IL-22对纤毛细胞的影响，并研究Hippo-Yes相关蛋白（YAP）信号传导对纤毛发生的影响。
方法：我们评估了从CRSwNP患者获得的鼻组织中IL-17A和IL-22的mRNA和蛋白质表达水平，并将其与健康对照进行了比较。为了进一步探索IL-17A和IL-22的影响，我们使用不同浓度（2ng/mL，10ng/mL,50ng/mL），在气液界面（ALI）培养物中持续28天。此外,我们使用抑制剂维替泊芬（VP）来研究IL-17A和IL-22是否通过Hippo-YAP途径对纤毛细胞发挥作用。
结果：CRSwNP中IL-17A和IL-22的mRNA和蛋白水平明显高于健康对照组，揭示了IL-17A和IL-22之间的强相关性。YAP在CRSwNP的纤毛细胞核中高表达，并与临床症状呈正相关。发现IL-17A和IL-22均可减少纤毛细胞的数量。IL-17A,但不是IL-22，通过破坏纤毛细胞基体的适当发育和对接来抑制纤毛生成，导致活动纤毛功能障碍。此外，随着纤毛细胞达到分化的最后阶段，纤毛细胞核内YAP的表达逐渐下降。然而,该过程仅被IL-17A阻碍。YAP抑制剂，比如Verteporfin,通过增加纤毛细胞的比例显着逆转了IL-17A的作用，抑制这些细胞中的核YAP表达，增强纤毛跳动频率。
结论：IL-17A和IL-22在CRSwNP的鼻上皮中过表达，这与上皮细胞分化受损有关。此外，已显示IL-17A通过激活YAP对活动纤毛的形态发生具有破坏性作用。
方法：我们评估了从CRSwNP患者获得的鼻组织中IL-17A和IL-22的mRNA和蛋白质表达水平，并将其与健康对照进行了比较。为了进一步探索IL-17A和IL-22的影响，我们使用不同浓度（2ng/mL，10ng/mL,50ng/mL），在气液界面（ALI）培养物中持续28天。此外,我们使用抑制剂维替泊芬（VP）来研究IL-17A和IL-22是否通过Hippo-YAP途径对纤毛细胞发挥作用。
结果：CRSwNP中IL-17A和IL-22的mRNA和蛋白水平明显高于健康对照组，揭示了IL-17A和IL-22之间的强相关性。YAP在CRSwNP的纤毛细胞核中高表达，并与临床症状呈正相关。发现IL-17A和IL-22均可减少纤毛细胞的数量。IL-17A,但不是IL-22，通过破坏纤毛细胞基体的适当发育和对接来抑制纤毛生成，导致活动纤毛功能障碍。此外，随着纤毛细胞达到分化的最后阶段，纤毛细胞核内YAP的表达逐渐下降。然而,该过程仅被IL-17A阻碍。YAP抑制剂，比如Verteporfin,通过增加纤毛细胞的比例显着逆转了IL-17A的作用，抑制这些细胞中的核YAP表达，增强纤毛跳动频率。
结论：IL-17A和IL-22在CRSwNP的鼻上皮中过表达，这与上皮细胞分化受损有关。此外，已显示IL-17A通过激活YAP对活动纤毛的形态发生具有破坏性作用。
