Morphological single cell profiling of the epithelial-mesenchymal transition
Morphological single cell profiling of the epithelial-mesenchymal transition
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DOI：
10.1039/c6ib00139d
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发表时间：
2016-01-01
影响因子：
2.5
通讯作者：
Wong, Ian Y.
中科院分区：
文献类型：
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作者：
Leggett, Susan E.;Sim, Jea Yun;Wong, Ian Y.
单细胞对生化处理的反应具有异质性，这使体外和体内实验分析变得复杂。特别是，应激扰动可能诱导上皮-间质转化（EMT），通过这种转化，致密、敏感的细胞采用了细长、抗性的表型。然而，传统的基于种群平均值的生物化学测量方法不能解决单细胞的异质性和可塑性问题。在这里，我们使用单细胞形态学的高含量成像来分类EMT后不同的表型亚群。我们首先通过主调节剂Snail在乳腺上皮细胞中描述了72小时内明确定义的EMT诱导。我们发现EMT与波形蛋白面积增加以及细胞核和细胞质的延长有关。这些形态学特征被整合到一个高斯混合模型中，该模型对上皮细胞和间充质细胞表型进行分类，准确率为bb0.92%。然后，我们将该分析应用于由较少控制的emt诱导刺激产生的异质群体，包括生长因子（tgf - β 1）、细胞密度和化疗药物（紫杉醇）。我们的定量单细胞方法有潜力筛选大量异质细胞群的许多类型的表型变异性，因此可能为靶向治疗的临床前评估提供预测分析。
Single cells respond heterogeneously to biochemical treatments, which can complicate the analysis of in vitro and in vivo experiments. In particular, stressful perturbations may induce the epithelial-mesenchymal transition (EMT), a transformation through which compact, sensitive cells adopt an elongated, resistant phenotype. However, classical biochemical measurements based on population averages over large numbers cannot resolve single cell heterogeneity and plasticity. Here, we use high content imaging of single cell morphology to classify distinct phenotypic subpopulations after EMT. We first characterize a well-defined EMT induction through the master regulator Snail in mammary epithelial cells over 72 h. We find that EMT is associated with increased vimentin area as well as elongation of the nucleus and cytoplasm. These morphological features were integrated into a Gaussian mixture model that classified epithelial and mesenchymal phenotypes with >92% accuracy. We then applied this analysis to heterogeneous populations generated from less controlled EMT-inducing stimuli, including growth factors (TGF-beta 1), cell density, and chemotherapeutics (Taxol). Our quantitative, single cell approach has the potential to screen large heterogeneous cell populations for many types of phenotypic variability, and may thus provide a predictive assay for the preclinical assessment of targeted therapeutics.