Asporin Promotes TGF-β-induced Lung Myofibroblast Differentiation by Facilitating Rab11-Dependent Recycling of TβRI
Asporin Promotes TGF-β-induced Lung Myofibroblast Differentiation by Facilitating Rab11-Dependent Recycling of TβRI
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DOI：
10.1165/rcmb.2021-0257oc
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发表时间：
2022-02-01
影响因子：
6.4
通讯作者：
Huang, Wenqi
中科院分区：
文献类型：
--
作者：
Huang, Shaojie;Lai, Xiaofan;Huang, Wenqi
特发性肺纤维化(IPF)是一种慢性进展性纤维性肺部疾病，病死率和发病率均较高。ASPN(Asporin)是富含亮氨酸的蛋白多糖家族中的一员，在组织损伤和再生中发挥重要作用。然而，ASPN在IPF中的确切病理生理作用及其分子机制尚不清楚。我们试图研究ASPN在肺纤维化发展中的作用以及靶向ASPN相关信号通路的治疗潜力。在我们的研究中，从基因表达总览数据库下载了三个微阵列数据集，并通过生物信息学分析筛选出差异表达的基因。Hub基因是从蛋白质-蛋白质相互作用网络中挑选出来的。检测肺纤维化模型小鼠肺组织中ASPN的表达，并通过体外转染ASPN shRNA载体，检测ASPN在转化生长因子-β/Smad信号转导中的作用。用生物素化方法检测ASPN敲除后细胞质膜TFG-β受体I(TβRI)和TβRI循环的变化。结果表明，肺纤维化模型小鼠肺组织中ASPN表达增加，且主要定位于α-SMA1肌成纤维细胞。体外实验证明，ASPN基因敲除通过调节TβRI的稳定性抑制了转化生长因子-β/Smad信号转导和肌成纤维细胞的分化。进一步的分子机制表明，ASPN基因敲除通过抑制TβRI以Rab11依赖的方式循环到细胞表面，从而抑制了TGF-β/Smad信号转导，并促进了溶酶体介导的TβRI的降解。总之，我们的发现为ASPN作为治疗肺纤维化的新的药理靶点的使用提供了重要的证据。
Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive fibrotic lung disease with high mortality and morbidity. ASPN (asporin), a member of the small leucine-rich proteoglycan family, plays crucial roles in tissue injury and regeneration. However, the precise pathophysiological role of ASPN and its molecular mechanisms in IPF remain unknown. We sought to investigate the role of ASPN during the development of pulmonary fibrosis and the therapeutic potential of targeting ASPN-related signaling pathways. In our study, three microarray datasets were downloaded from the Gene Expression Omnibus database, and differentially expressed genes were screened out by bioinformatic analysis. Hub genes were selected from the protein-protein interaction network. ASPN was examined in lung tissues from pulmonary fibrosis mouse models, and the role of ASPN in transforming growth factor (TGF)-beta/Smad signaling was determined by transfection with ASPN shRNA vectors in vitro. Biotinylation assays were conducted to measure plasma membrane TFG-beta receptor I (T beta RI) and T beta RI recycling after ASPN knockdown. The results showed ASPN expression was increased in the lungs of pulmonary fibrosis mouse models, and ASPN was primarily localized in alpha-SMA1 myofibroblasts. In vitro experiments proved that ASPN knockdown inhibited TGF-beta/Smad signaling and myofibroblast differentiation by regulating the stability of T beta RI. Further molecular mechanisms revealed that ASPN knockdown inhibited TGF-beta/Smad signaling by suppressing recycling of T beta RI to the cell surface in a Rab11-dependent manner and facilitated lysosome-mediated degradation of T beta RI. In conclusion, our findings provide important evidence for the use of ASPN as a novel pharmacological target for treating pulmonary fibrosis.