Spatial and temporal dynamics of RhoA activities of single breast tumor cells in a 3D environment revealed by a machine learning-assisted FRET technique.
Spatial and temporal dynamics of RhoA activities of single breast tumor cells in a 3D environment revealed by a machine learning-assisted FRET technique.
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DOI：
10.1016/j.yexcr.2021.112939
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发表时间：
2022-01-15
影响因子：
3.7
通讯作者：
Wu M
中科院分区：
文献类型：
--
作者：
Cheung BCH;Hodgson L;Segall JE;Wu M
癌细胞的特征之一是它们在细胞外基质(ECM)内迁移的特殊能力，以进入循环系统，这是癌症转移的关键步骤。RhoA是一种小的GTP酶，在细胞迁移过程中是一个关键的分子开关，在肌动蛋白收缩和板脂突起之间切换。目前对RhoA在细胞迁移中的活性的了解在很大程度上源于使用FRET生物传感器对二维(2D)衬底上的细胞进行的研究。越来越多的证据表明，细胞在更具生理相关性的三维(3D)环境中表现不同。然而，荧光成像的低信噪比阻碍了对三维RhoA活性的研究。在本文中，我们提出了一种FRET技术与机器学习辅助的细胞分割方法相结合，使用RhoA生物传感器来跟踪单个乳腺肿瘤细胞(MDA-MB-231)在3D和2D环境中迁移的时空动态。我们发现，与3D胶原基质相比，在2D纤维连接蛋白涂层玻璃上迁移的单个细胞的RhoA活性沿细胞长轴的极化程度更高。特别是，与3D相比，2D中细胞的RhoA活动在迁移过程中表现出明显的从前到后和从后到前的运动。最后，不管维度如何，RhoA极化与细胞形状之间存在适度的相关性。
One of the hallmarks of cancer cells is their exceptional ability to migrate within the extracellular matrix (ECM) for gaining access to the circulatory system, a critical step of cancer metastasis. RhoA, a small GTPase, is known to be a key molecular switch that toggles between actomyosin contractility and lamellipodial protrusion during cell migration. Current understanding of RhoA activity in cell migration has been largely derived from studies of cells plated on a two-dimensional (2D) substrate using a FRET biosensor. There has been increasing evidence that cells behave differently in a more physiologically relevant three-dimensional (3D) environment. However, studies of RhoA activities in 3D have been hindered by low signal-to-noise ratio in fluorescence imaging. In this paper, we present a FRET technique in conjunction with a machine learning-assisted cell segmentation method to follow the spatiotemporal dynamics of RhoA activities of single breast tumor cells (MDA-MB-231) migrating in a 3D as well as a 2D environment using a RhoA biosensor. We found that RhoA activity is more polarized along the long axis of the cell for single cells migrating on 2D fibronectin-coated glass versus those embedded in 3D collagen matrices. In particular, RhoA activities of cells in 2D exhibit a distinct front-to-back and back-to-front movement during migration in contrast to those in 3D. Finally, regardless of dimensionality, RhoA polarization is found to be moderately correlated with cell shape.
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影响因子：
3.4
作者：
Costigliola, Nancy;Kapustina, Maryna T.;Jacobson, Ken
通讯作者：
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影响因子：
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作者：
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影响因子：
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DOI：
10.1007/978-1-4939-8612-5_6
发表时间：
2018-01-01
期刊：
RHO GTPASES: METHODS AND PROTOCOLS, 2ND EDITION
影响因子：
--
作者：
Bell, George R. R.;Natwick, Dean E.;Collins, Sean R.
通讯作者：
Collins, Sean R.
影响因子：
14.8
作者：
Hodgson L;Spiering D;Sabouri-Ghomi M;Dagliyan O;DerMardirossian C;Danuser G;Hahn KM
通讯作者：
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