Abstract：
BACKGROUND: The role of IL-13 on the airway epithelium in severe asthma leading to airway remodeling remains poorly understood.
OBJECTIVE: To study IL-13 induced airway remodeling on goblet cells and cilia in the airway epithelium in severe asthma and the impact of an anti-IL4Rα antibody, dupilumab, in vitro.
METHODS: Quantitative CT (qCT) lungs and endobronchial biopsies and brushings were obtained in 51 participants (22 severe, 11 non-severe asthma and 18 healthy participants) in the Severe Asthma Research Program (SARPIII) and measured for mucin and cilia related proteins. Epithelial cells were differentiated in air-liquid interphase (ALI) with IL-13 +/-dupilumab and assessed for mucin, cilia, cilia beat frequency (CBF) and epithelial integrity (transepithelial electrical resistance, TEER).
RESULTS: Increased Muc5AC (Δ+263.2±92.7 lums/EpiArea) and decreased ciliated cells (Δ-0.07±0.03 Foxj1+cells/EpiArea) were observed in biopsies from severe asthma when compared to healthy (p<0.01 and p=0.047 respectively). RNAseq of epithelial cell brushes confirmed a Muc5AC increase with a decrease in a 5-gene cilia-related mean in severe asthma compared to healthy (all p<0.05). IL-13 (5 ng/mL) differentiated ALI cultures of healthy and asthmatic (severe and non-severe participants) increased Muc5AC, decreased cilia (α-acytl-tubulin) in healthy (Δ+6.5±1.5%, Δ-14.1±2.7%; all p<0.001 respectively) and asthma (Δ+4.4±2.5%, Δ-13.1±2.7%; p=0.084, p<0.001 respectively); decreased epithelial integrity (TEER) in healthy (-140.9±21.3 [ohms], p<0.001) while decreasing CBF in asthma (Δ-4.4±1.7 [Hz], p<0.01). When dupilumab was added to ALI with IL-13, there was no significant decrease in Mu5AC but there was restoration of cilia in healthy and asthma participants (absolute increase of 67.5% and 32.5% cilia, all p<0.05 respectively) while CBF increased (Δ+3.6±1.1 [Hz], p<0.001) and TEER decreased (only in asthma Δ-37.8±16.2 [ohms] p<0.05).
CONCLUSIONS: IL-13 drives features of airway remodeling in severe asthma which are partially reversed by inhibiting IL-4Rα receptor in vitro.
OBJECTIVE: To study IL-13 induced airway remodeling on goblet cells and cilia in the airway epithelium in severe asthma and the impact of an anti-IL4Rα antibody, dupilumab, in vitro.
METHODS: Quantitative CT (qCT) lungs and endobronchial biopsies and brushings were obtained in 51 participants (22 severe, 11 non-severe asthma and 18 healthy participants) in the Severe Asthma Research Program (SARPIII) and measured for mucin and cilia related proteins. Epithelial cells were differentiated in air-liquid interphase (ALI) with IL-13 +/-dupilumab and assessed for mucin, cilia, cilia beat frequency (CBF) and epithelial integrity (transepithelial electrical resistance, TEER).
RESULTS: Increased Muc5AC (Δ+263.2±92.7 lums/EpiArea) and decreased ciliated cells (Δ-0.07±0.03 Foxj1+cells/EpiArea) were observed in biopsies from severe asthma when compared to healthy (p<0.01 and p=0.047 respectively). RNAseq of epithelial cell brushes confirmed a Muc5AC increase with a decrease in a 5-gene cilia-related mean in severe asthma compared to healthy (all p<0.05). IL-13 (5 ng/mL) differentiated ALI cultures of healthy and asthmatic (severe and non-severe participants) increased Muc5AC, decreased cilia (α-acytl-tubulin) in healthy (Δ+6.5±1.5%, Δ-14.1±2.7%; all p<0.001 respectively) and asthma (Δ+4.4±2.5%, Δ-13.1±2.7%; p=0.084, p<0.001 respectively); decreased epithelial integrity (TEER) in healthy (-140.9±21.3 [ohms], p<0.001) while decreasing CBF in asthma (Δ-4.4±1.7 [Hz], p<0.01). When dupilumab was added to ALI with IL-13, there was no significant decrease in Mu5AC but there was restoration of cilia in healthy and asthma participants (absolute increase of 67.5% and 32.5% cilia, all p<0.05 respectively) while CBF increased (Δ+3.6±1.1 [Hz], p<0.001) and TEER decreased (only in asthma Δ-37.8±16.2 [ohms] p<0.05).
CONCLUSIONS: IL-13 drives features of airway remodeling in severe asthma which are partially reversed by inhibiting IL-4Rα receptor in vitro.
摘要：
背景：在严重哮喘导致气道重塑的过程中，IL-13对气道上皮的作用仍知之甚少。
目的：研究IL-13诱导的重度哮喘气道上皮杯状细胞和纤毛的气道重塑以及抗IL4Rα抗体的影响，dupilumab,在体外。
方法：在51名参与者中获得了定量CT（qCT）肺和支气管内活检和刷检（22名重度，11名非重度哮喘和18名健康参与者）参加了重度哮喘研究计划（SARPIII），并测量了粘蛋白和纤毛相关蛋白。用IL-13+/-dupilumab在气液间期（ALI）中分化上皮细胞，并评估粘蛋白，纤毛,纤毛搏动频率(CBF)和上皮完整性(跨上皮电阻，TEER).
结果：在重度哮喘的活检中观察到Muc5AC增加（Δ263.2±92.7lums/EpiArea）和纤毛细胞减少（Δ-0.07±0.03Foxj1细胞/EpiArea）与健康相比（分别为p<0.01和p=0.047）。上皮细胞刷的RNAseq证实，与健康相比，重度哮喘中Muc5AC增加，5基因纤毛相关平均值降低（所有p<0.05）。IL-13（5ng/mL）分化的健康和哮喘（重度和非重度参与者）的ALI培养物增加Muc5AC，健康的纤毛（α-acytl-微管蛋白）减少（Δ6.5±1.5％，Δ-14.1±2.7％；所有p分别<0.001）和哮喘（Δ4.4±2.5％，Δ-13.1±2.7％；p=0.084，p<0.001）；健康患者的上皮完整性（TEER）降低（-140.9±21.3[ohms]，p<0.001），同时降低哮喘患者的CBF（Δ-4.4±1.7[Hz]，p<0.01)。当dupilumab与IL-13一起加入ALI时，Mu5AC没有显着降低，但健康和哮喘参与者的纤毛恢复（绝对增加67.5％和32.5％纤毛，所有p分别<0.05)，而CBF增加(Δ+3.6±1.1[Hz]，p<0.001）和TEER降低（仅在哮喘中Δ-37.8±16.2[ohms]p<0.05）。
结论：IL-13驱动重度哮喘气道重塑的特征，其在体外通过抑制IL-4Rα受体而部分逆转。
目的：研究IL-13诱导的重度哮喘气道上皮杯状细胞和纤毛的气道重塑以及抗IL4Rα抗体的影响，dupilumab,在体外。
方法：在51名参与者中获得了定量CT（qCT）肺和支气管内活检和刷检（22名重度，11名非重度哮喘和18名健康参与者）参加了重度哮喘研究计划（SARPIII），并测量了粘蛋白和纤毛相关蛋白。用IL-13+/-dupilumab在气液间期（ALI）中分化上皮细胞，并评估粘蛋白，纤毛,纤毛搏动频率(CBF)和上皮完整性(跨上皮电阻，TEER).
结果：在重度哮喘的活检中观察到Muc5AC增加（Δ263.2±92.7lums/EpiArea）和纤毛细胞减少（Δ-0.07±0.03Foxj1细胞/EpiArea）与健康相比（分别为p<0.01和p=0.047）。上皮细胞刷的RNAseq证实，与健康相比，重度哮喘中Muc5AC增加，5基因纤毛相关平均值降低（所有p<0.05）。IL-13（5ng/mL）分化的健康和哮喘（重度和非重度参与者）的ALI培养物增加Muc5AC，健康的纤毛（α-acytl-微管蛋白）减少（Δ6.5±1.5％，Δ-14.1±2.7％；所有p分别<0.001）和哮喘（Δ4.4±2.5％，Δ-13.1±2.7％；p=0.084，p<0.001）；健康患者的上皮完整性（TEER）降低（-140.9±21.3[ohms]，p<0.001），同时降低哮喘患者的CBF（Δ-4.4±1.7[Hz]，p<0.01)。当dupilumab与IL-13一起加入ALI时，Mu5AC没有显着降低，但健康和哮喘参与者的纤毛恢复（绝对增加67.5％和32.5％纤毛，所有p分别<0.05)，而CBF增加(Δ+3.6±1.1[Hz]，p<0.001）和TEER降低（仅在哮喘中Δ-37.8±16.2[ohms]p<0.05）。
结论：IL-13驱动重度哮喘气道重塑的特征，其在体外通过抑制IL-4Rα受体而部分逆转。
