TFEB信号通路减弱nlrp3驱动的严重哮喘炎症反应
2022/02/28
摘要
背景：由循环和肺部驻留的单核细胞介导的nlrp3驱动的炎症反应是哮喘发病的关键因素。自噬抑制哮喘模型中NLRP3诱导的单核细胞激活。然而，自噬及其主要调控因子转录因子EB (TFEB)对人哮喘单核细胞反应的影响仍未被探索。在这里，我们研究了自噬和TFEB信号通路的激活是否抑制哮喘个体的炎症单核细胞反应。
方法：采用LPS/ATP刺激哮喘患者(n = 83)和健康对照组(n = 46)的外周血CD14+单核细胞，诱导NLRP3活化，并加入或不加入自噬诱导物雷帕霉素。检测ASC斑点、caspase-1激活、IL-1β和IL-18水平、线粒体功能、ROS释放和mTORC1信号。通过LC3点状细胞形成、p62/SQSTM1降解和TFEB激活来评估自噬。在严重哮喘(SA)模型中，我们使用NLRP3−/−小鼠和/或给药MCC950研究了NLRP3信号通路的作用，并使用骨髓特异性TFEB过表达小鼠或给药TFEB激活剂海藻糖研究了TFEB激活的影响。
结果：我们观察到NLRP3炎症小体激活增加，伴随与哮喘严重程度相关的循环单核细胞自噬受损。SA患者还表现为线粒体功能障碍和ROS积累。自噬无法抑制nlrp3驱动的单核细胞反应，这是由于TFEB活化缺陷和过度的mTORC1信号通路。阻断NLRP3可抑制炎症细胞因子释放和相关气道疾病。TFEB激活可恢复受损的自噬，减轻nlrp3驱动的肺部炎症，并改善SA表型。
结论：我们的研究揭示了TFEB介导的单核细胞炎症反应重编程的关键作用，提高了这一途径可用于SA治疗的前景。
TFEB signaling attenuates NLRP3-driven inflammatory responses in severe asthma
Theofani E, Semitekolou M, Samitas K, et al.
Abstract
BACKGROUND：NLRP3-driven inflammatory responses by circulating and lung-resident monocytes are critical drivers of asthma pathogenesis. Autophagy restrains NLRP3- induced monocyte activation in asthma models. Yet, the effects of autophagy and its master regulator, transcription factor EB (TFEB), on monocyte responses in human asthma remain unexplored. Here, we investigated whether activation of autophagy and TFEB signaling suppress inflammatory monocyte responses in asthmatic individuals.
METHODS：Peripheral blood CD14+ monocytes from asthmatic patients (n = 83) and healthy controls (n = 46) were stimulated with LPS/ATP to induce NLRP3 activation with or without the autophagy inducer, rapamycin. ASC specks, caspase-1 activation, IL-1β and IL-18 levels, mitochondrial function, ROS release, and mTORC1 signaling were examined. Autophagy was evaluated by LC3 puncta formation, p62/SQSTM1 degradation and TFEB activation. In a severe asthma (SA) model, we investigated the role of NLRP3 signaling using Nlrp3−/− mice and/or MCC950 administration, and the effects of TFEB activation using myeloid-specific TFEB-overexpressing mice or administration of the TFEB activator, trehalose.
RESULTS：We observed increased NLRP3 inflammasome activation, concomitant with impaired autophagy in circulating monocytes that correlated with asthma severity. SA patients also exhibited mitochondrial dysfunction and ROS accumulation. Autophagy failed to inhibit NLRP3-driven monocyte responses, due to defective TFEB activation and excessive mTORC1 signaling. NLRP3 blockade restrained inflammatory cytokine release and linked airway disease. TFEB activation restored impaired autophagy, attenuated NLRP3-driven pulmonary inflammation, and ameliorated SA phenotype.
CONCLUSIONS：Our studies uncover a crucial role for TFEB-mediated reprogramming of monocyte inflammatory responses, raising the prospect that this pathway can be therapeutically harnessed for the management of SA.
背景：由循环和肺部驻留的单核细胞介导的nlrp3驱动的炎症反应是哮喘发病的关键因素。自噬抑制哮喘模型中NLRP3诱导的单核细胞激活。然而，自噬及其主要调控因子转录因子EB (TFEB)对人哮喘单核细胞反应的影响仍未被探索。在这里，我们研究了自噬和TFEB信号通路的激活是否抑制哮喘个体的炎症单核细胞反应。
方法：采用LPS/ATP刺激哮喘患者(n = 83)和健康对照组(n = 46)的外周血CD14+单核细胞，诱导NLRP3活化，并加入或不加入自噬诱导物雷帕霉素。检测ASC斑点、caspase-1激活、IL-1β和IL-18水平、线粒体功能、ROS释放和mTORC1信号。通过LC3点状细胞形成、p62/SQSTM1降解和TFEB激活来评估自噬。在严重哮喘(SA)模型中，我们使用NLRP3−/−小鼠和/或给药MCC950研究了NLRP3信号通路的作用，并使用骨髓特异性TFEB过表达小鼠或给药TFEB激活剂海藻糖研究了TFEB激活的影响。
结果：我们观察到NLRP3炎症小体激活增加，伴随与哮喘严重程度相关的循环单核细胞自噬受损。SA患者还表现为线粒体功能障碍和ROS积累。自噬无法抑制nlrp3驱动的单核细胞反应，这是由于TFEB活化缺陷和过度的mTORC1信号通路。阻断NLRP3可抑制炎症细胞因子释放和相关气道疾病。TFEB激活可恢复受损的自噬，减轻nlrp3驱动的肺部炎症，并改善SA表型。
结论：我们的研究揭示了TFEB介导的单核细胞炎症反应重编程的关键作用，提高了这一途径可用于SA治疗的前景。
（中日友好医院呼吸与危重症医学科 李春晓 摘译 林江涛 审校）
（Allergy, 2022.）
（Allergy, 2022.）
TFEB signaling attenuates NLRP3-driven inflammatory responses in severe asthma
Theofani E, Semitekolou M, Samitas K, et al.
Abstract
BACKGROUND：NLRP3-driven inflammatory responses by circulating and lung-resident monocytes are critical drivers of asthma pathogenesis. Autophagy restrains NLRP3- induced monocyte activation in asthma models. Yet, the effects of autophagy and its master regulator, transcription factor EB (TFEB), on monocyte responses in human asthma remain unexplored. Here, we investigated whether activation of autophagy and TFEB signaling suppress inflammatory monocyte responses in asthmatic individuals.
METHODS：Peripheral blood CD14+ monocytes from asthmatic patients (n = 83) and healthy controls (n = 46) were stimulated with LPS/ATP to induce NLRP3 activation with or without the autophagy inducer, rapamycin. ASC specks, caspase-1 activation, IL-1β and IL-18 levels, mitochondrial function, ROS release, and mTORC1 signaling were examined. Autophagy was evaluated by LC3 puncta formation, p62/SQSTM1 degradation and TFEB activation. In a severe asthma (SA) model, we investigated the role of NLRP3 signaling using Nlrp3−/− mice and/or MCC950 administration, and the effects of TFEB activation using myeloid-specific TFEB-overexpressing mice or administration of the TFEB activator, trehalose.
RESULTS：We observed increased NLRP3 inflammasome activation, concomitant with impaired autophagy in circulating monocytes that correlated with asthma severity. SA patients also exhibited mitochondrial dysfunction and ROS accumulation. Autophagy failed to inhibit NLRP3-driven monocyte responses, due to defective TFEB activation and excessive mTORC1 signaling. NLRP3 blockade restrained inflammatory cytokine release and linked airway disease. TFEB activation restored impaired autophagy, attenuated NLRP3-driven pulmonary inflammation, and ameliorated SA phenotype.
CONCLUSIONS：Our studies uncover a crucial role for TFEB-mediated reprogramming of monocyte inflammatory responses, raising the prospect that this pathway can be therapeutically harnessed for the management of SA.
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肺部炎症对人和小鼠胰岛素抵抗的不同影响
