Fibronectin on the Surface of Extracellular Vesicles Mediates Fibroblast Invasion
Fibronectin on the Surface of Extracellular Vesicles Mediates Fibroblast Invasion
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DOI：
10.1165/rcmb.2018-0062oc
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发表时间：
2019-03-01
影响因子：
6.4
通讯作者：
Thannicka, Victor J.
中科院分区：
文献类型：
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作者：
Chanda, Diptiman;Otoupalova, Eva;Thannicka, Victor J.
细胞外囊泡（EV）是核内体和质膜衍生的纳米大小的囊泡，参与细胞间信号传导。虽然EV货物可能通过多种机制发出信号，但EV表面上的信号传导组分如何介导细胞信号传导还不太清楚。在这项研究中，我们发现成纤维细胞衍生的EV在囊泡表面携带纤连蛋白，这一点通过基于质谱的蛋白质组学（所有理论质谱的顺序窗口采集）和流式细胞术分析得到证实。经历复制性衰老或转化生长因子β 1诱导的衰老的成纤维细胞和从患有年龄相关性肺病、特发性肺纤维化的人类受试者中分离的成纤维细胞分泌的EV数量高于其各自的对照。成纤维细胞衍生的EV在受体成纤维细胞中诱导侵袭性表型。这种侵袭性成纤维细胞表型依赖于纤维连接蛋白的EV表面定位、与纤维连接蛋白受体α 5 β 1整合素的相互作用以及涉及粘着斑激酶和Src家族激酶的侵袭相关信号通路的激活。细胞上清液中的EV，未结合细胞外基质，能够介导受体成纤维细胞上的侵袭信号，支持EV表面纤连蛋白与受体细胞质膜的直接相互作用。总之，这些研究揭示了成纤维细胞侵袭的EV信号传导的新机制，其可能与纤维化疾病和癌症的发病机制相关。
Extracellular vesicles (EVs) are endosome and plasma membrane-derived nano-sized vesicles that participate in intercellular signaling. Although EV cargo may signal via multiple mechanisms, how signaling components on the surface of EVs mediate cellular signaling is less well understood. In this study, we show that fibroblast-derived EVs carry fibronectin on the vesicular surface, as evidenced by mass spectrometry-based proteomics (Sequential Window Acquisition of all Theoretical Mass Spectra) and flow-cytometric analyses. Fibroblasts undergoing replicative senescence or transforming growth factor beta 1-induced senescence and fibroblasts isolated from human subjects with an age-related lung disorder, idiopathic pulmonary fibrosis, secreted higher numbers of EVs than their respective controls. Fibroblast-derived EVs induced an invasive phenotype in recipient fibroblasts. This invasive fibroblast phenotype was dependent on EV surface localization of fibronectin, interaction with the fibronectin receptor alpha 5 beta 1 integrin, and activation of invasion-associated signaling pathways involving focal adhesion kinase and Src family kinases. EVs in the cellular supernatant, unbound to the extracellular matrix, were capable of mediating invasion signaling on recipient fibroblasts, supporting a direct interaction of EV surface fibronectin with the plasma membrane of recipient cells. Together, these studies uncover a novel mechanism of EV signaling of fibroblast invasion that may be relevant in the pathogenesis of fibrotic diseases and cancer.