Directional secretomes reflect polarity-specific functions in an in vitro model of human bronchial epithelium.
Directional secretomes reflect polarity-specific functions in an in vitro model of human bronchial epithelium.
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DOI：
10.1165/rcmb.2013-0188oc
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发表时间：
2013-09
影响因子：
6.4
通讯作者：
D. Pillai;Binu-John Sankoorikal;Eric Johnson;A. N. Seneviratne;Jessica Zurko;K. Brown;Y. Hathout;M. Rose
中科院分区：
文献类型：
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作者：
D. Pillai;Binu-John Sankoorikal;Eric Johnson;A. N. Seneviratne;Jessica Zurko;K. Brown;Y. Hathout;M. Rose
传导气道上皮的极性负责其定向分泌。这是肺完整性和功能的基本特征，决定了外部环境（顶端）和上皮下结构（基底外侧）之间的相互作用。在体外人支气管上皮细胞（HBE）分化模型中确定定向分泌组可以为肺生物学和肺部疾病带来有价值的见解。正常原代HBE细胞（n = 3）分化为呼吸道上皮。处理顶部和基底侧分泌物（24小时）用于蛋白质组分析和途径分析。在所有HBE培养物的分泌物中共鉴定出243种蛋白质。其中，51%被归类为分泌蛋白，包括真分泌蛋白（36%）和外泌体蛋白（15%）。仔细检查显示，所有供体中69种顶端蛋白和13种基底外侧蛋白的分泌一致，25种蛋白的分泌差异。在一式三份的独立实验中，使用蛋白质印迹或ELISA验证顶端分泌物中的膜联蛋白A4和基底外侧分泌物中的桥粒芯糖蛋白-2的表达。据我们所知，这是首次在体外分化HBE模型中定义顶侧和基底侧分泌体的研究。这些数据表明，上皮极性指导蛋白质分泌与不同模式的生物过程的顶端和基底侧表面，这是符合正常的支气管上皮稳态功能。将这种体外定向分泌组模型应用于肺部疾病可能会阐明其分子病理生理学，并有助于确定潜在的治疗靶点。
The polarity of the conducting airway epithelium is responsible for its directional secretion. This is an essential characteristic of lung integrity and function that dictates interactions between the external environment (apical) and subepithelial structures (basolateral). Defining the directional secretomes in the in vitro human bronchial epithelial (HBE) differentiated model could bring valuable insights into lung biology and pulmonary diseases. Normal primary HBE cells (n = 3) were differentiated into respiratory tract epithelium. Apical and basolateral secretions (24 h) were processed for proteome profiling and pathway analysis. A total of 243 proteins were identified in secretions from all HBE cultures combined. Of these, 51% were classified as secreted proteins, including true secreted proteins (36%) and exosomal proteins (15%). Close examination revealed consistent secretion of 69 apical proteins and 13 basolateral proteins and differential secretion of 25 proteins across all donors. Expression of Annexin A4 in apical secretions and Desmoglein-2 in basolateral secretions was validated using Western blot or ELISA in triplicate independent experiments. To the best of our knowledge, this is the first study defining apical and basolateral secretomes in the in vitro differentiated HBE model. The data demonstrate that epithelial polarity directs protein secretion with different patterns of biological processes to the apical and basolateral surfaces that are consistent with normal bronchial epithelium homeostatic functions. Applying this in vitro directional secretome model to lung diseases may elucidate their molecular pathophysiology and help define potential therapeutic targets.