KIF5A transports collagen vesicles of myofibroblasts during pleural fibrosis.
KIF5A transports collagen vesicles of myofibroblasts during pleural fibrosis.
复制标题
DOI：
10.1038/s41598-017-04437-7
复制
发表时间：
2017-07-04
影响因子：
4.6
通讯作者：
Ikebe M
中科院分区：
文献类型：
--
作者：
Kamata H;Tsukasaki Y;Sakai T;Ikebe R;Wang J;Jeffers A;Boren J;Owens S;Suzuki T;Higashihara M;Idell S;Tucker TA;Ikebe M
纤维化涉及组织中细胞外基质蛋白的产生，并且通常在损伤或创伤之前。在胸膜纤维化中，过量胶原沉积导致胸膜增厚、僵硬增加和肺功能受损。肌成纤维细胞是增加胶原沉积的原因，然而，含有用于分泌的囊泡的前胶原的转运的分子机制是未知的。在这里，我们研究了驱动蛋白对人胸膜间皮细胞（HPMCs）中含胶原-1（Col-1）的囊泡运输的作用。在许多货物运输驱动蛋白中，KIF 5A在TGF-β诱导的间皮-间充质转化（MesoMT）期间显著上调。使用超分辨结构照明显微镜和DUO-Link技术，我们发现KIF 5A与Col-1含有囊泡共定位。KIF 5A敲低显著减少Col-1分泌，并减弱TGF-β诱导的Col-1在细胞周边定位的增加。活细胞成像显示含有GFP-KIF 5A和mCherry-Col-1的囊泡一起移动。Kymography显示这些分子以0.56 μm/sec的平均速度连续运动，表明运动是定向的而不是扩散限制的过程。此外，在炭黑博莱霉素小鼠模型中，KIF 5A与Col-1和α-平滑肌肌动蛋白一起在胸膜增厚中显著上调沿着。这些结果支持我们的假设，KIF 5A是负责胶原蛋白的运输和分泌从HPMCs。
Fibrosis involves the production of extracellular matrix proteins in tissues and is often preceded by injury or trauma. In pleural fibrosis excess collagen deposition results in pleural thickening, increased stiffness and impaired lung function. Myofibroblasts are responsible for increased collagen deposition, however the molecular mechanism of transportation of procollagen containing vesicles for secretion is unknown. Here, we studied the role of kinesin on collagen-1 (Col-1) containing vesicle transportation in human pleural mesothelial cells (HPMCs). Among a number of cargo transporting kinesins, KIF5A was notably upregulated during TGF-β induced mesothelial-mesenchymal transition (MesoMT). Using superresolution structured illumination microscopy and the DUO-Link technique, we found that KIF5A colocalized with Col-1 containing vesicles. KIF5A knock-down significantly reduced Col-1 secretion and attenuated TGF-β induced increment in Col-1 localization at cell peripheries. Live cell imaging revealed that GFP-KIF5A and mCherry-Col-1 containing vesicles moved together. Kymography showed that these molecules continuously move with a mean velocity of 0.56 μm/sec, suggesting that the movement is directional but not diffusion limited process. Moreover, KIF5A was notably upregulated along with Col-1 and α-smooth muscle actin in pleural thickening in the carbon-black bleomycin mouse model. These results support our hypothesis that KIF5A is responsible for collagen transportation and secretion from HPMCs.