Program in Cell, Molecular and Developmental Biology, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts, USA.
PLoS One. 2013;8(2):e55911. doi: 10.1371/journal.pone.0055911. Epub 2013 Feb 8.
Epithelial-mesenchymal interactions play a crucial role in branching morphogenesis, but very little is known about how endothelial cells contribute to this process. Here, we examined how anti-angiogenic miR-221 and pro-angiogenic miR-130a affect airway and vascular development in the fetal lungs. Lung-specific effects of miR-130a and miR-221 were studied in mouse E14 whole lungs cultured for 48 hours with anti-miRs or mimics to miR-130a and miR-221. Anti-miR 221 treated lungs had more distal branch generations with increased Hoxb5 and VEGFR2 around airways. Conversely, mimic 221 treated lungs had reduced airway branching, dilated airway tips and decreased Hoxb5 and VEGFR2 in mesenchyme. Anti-miR 130a treatment led to reduced airway branching with increased Hoxa5 and decreased VEGFR2 in the mesenchyme. Conversely, mimic 130a treated lungs had numerous finely arborized branches extending into central lung regions with diffusely localized Hoxa5 and increased VEGFR2 in the mesenchyme. Vascular morphology was analyzed by GSL-B4 (endothelial cell-specific lectin) immunofluorescence. Observed changes in airway morphology following miR-221 inhibition and miR-130a enhancement were mirrored by changes in vascular plexus formation around the terminal airways. Mouse fetal lung endothelial cells (MFLM-91U) were used to study microvascular cell behavior. Mimic 221 treatment resulted in reduced tube formation and cell migration, where as the reverse was observed with mimic 130a treatment. From these data, we conclude that miR-221 and miR-130a have opposing effects on airway and vascular morphogenesis of the developing lung.
上皮-间充质相互作用在分支形态发生中起着至关重要的作用，但关于内皮细胞如何促进这一过程知之甚少。在这里，我们研究了抗血管生成 miR-221 和促血管生成 miR-130a 如何影响胎儿肺中的气道和血管发育。在培养了 48 小时的 E14 全肺中，用抗 miR-130a 和 miR-221 的抗 miR 或 mimics 研究了 miR-130a 和 miR-221 的肺特异性作用。用抗 miR-221 处理的肺有更多的远端分支世代，气道周围的 Hoxb5 和 VEGFR2 增加。相反，模拟 221 处理的肺气道分支减少，气道尖端扩张，间充质中的 Hoxb5 和 VEGFR2 减少。用抗 miR-130a 处理导致气道分支减少，间充质中的 Hoxa5 增加和 VEGFR2 减少。相反，模拟 130a 处理的肺有许多精细分支延伸到中央肺区，间充质中的 Hoxa5 弥散定位增加，VEGFR2 增加。通过 GSL-B4（内皮细胞特异性凝集素）免疫荧光分析血管形态。miR-221 抑制和 miR-130a 增强后气道形态的变化与终末气道周围血管丛形成的变化相吻合。使用小鼠胎儿肺内皮细胞（MFLM-91U）研究微血管细胞行为。模拟 221 处理导致管形成和细胞迁移减少，而模拟 130a 处理则相反。从这些数据中，我们得出结论，miR-221 和 miR-130a 对发育中肺的气道和血管形态发生有相反的影响。
