Comprehensive Pneumology Center, Helmholtz Center Munich, Ludwig Maximilians University Munich, University Hospital Grosshadern, Member of the German Center for Lung Research (DZL), Munich, Germany.
Asklepios Clinics, Gauting, Germany.
Eur Respir J. 2015 Oct;46(4):1150-66. doi: 10.1183/09031936.00183214. Epub 2015 Apr 30.
Chronic obstructive pulmonary disease (COPD) is characterised by a progressive loss of lung tissue. Inducing repair processes within the adult diseased lung is of major interest and Wnt/β-catenin signalling represents a promising target for lung repair. However, the translation of novel therapeutic targets from model systems into clinical use remains a major challenge.We generated murine and patient-derived three-dimensional (3D) ex vivo lung tissue cultures (LTCs), which closely mimic the 3D lung microenvironment in vivo. Using two well-known glycogen synthase kinase-3β inhibitors, lithium chloride (LiCl) and CHIR 99021 (CT), we determined Wnt/β-catenin-driven lung repair processes in high spatiotemporal resolution using quantitative PCR, Western blotting, ELISA, (immuno)histological assessment, and four-dimensional confocal live tissue imaging.Viable 3D-LTCs exhibited preserved lung structure and function for up to 5 days. We demonstrate successful Wnt/β-catenin signal activation in murine and patient-derived 3D-LTCs from COPD patients. Wnt/β-catenin signalling led to increased alveolar epithelial cell marker expression, decreased matrix metalloproteinase-12 expression, as well as altered macrophage activity and elastin remodelling. Importantly, induction of surfactant protein C significantly correlated with disease stage (per cent predicted forced expiratory volume in 1 s) in patient-derived 3D-LTCs.Patient-derived 3D-LTCs represent a valuable tool to analyse potential targets and drugs for lung repair. Enhanced Wnt/β-catenin signalling attenuated pathological features of patient-derived COPD 3D-LTCs.
慢性阻塞性肺疾病（COPD）的特征是肺组织的进行性丧失。在成人患病肺中诱导修复过程是非常重要的，Wnt/β-catenin 信号代表了肺修复的一个有前途的靶点。然而，将新的治疗靶点从模型系统转化为临床应用仍然是一个主要的挑战。我们生成了鼠和患者来源的三维（3D）离体肺组织培养物（LTC），这些培养物非常类似于体内的 3D 肺微环境。使用两种众所周知的糖原合酶激酶-3β抑制剂氯化锂（LiCl）和 CHIR 99021（CT），我们使用定量 PCR、Western blot、ELISA、（免疫）组织学评估和四维共聚焦活组织成像，以高时空分辨率确定 Wnt/β-catenin 驱动的肺修复过程。可行的 3D-LTC 可在长达 5 天的时间内保持肺结构和功能。我们证明了在鼠和来自 COPD 患者的患者来源的 3D-LTC 中成功激活了 Wnt/β-catenin 信号。Wnt/β-catenin 信号导致肺泡上皮细胞标志物表达增加，基质金属蛋白酶-12 表达减少，以及巨噬细胞活性和弹性蛋白重塑改变。重要的是，表面活性剂蛋白 C 的诱导与患者来源的 3D-LTC 中的疾病阶段（预计用力呼气量的百分比）显著相关。患者来源的 3D-LTC 是分析潜在靶点和肺修复药物的有价值的工具。增强的 Wnt/β-catenin 信号减弱了患者来源的 COPD 3D-LTC 的病理性特征。
