Abstract
In order to metastasise, triple negative breast cancer (TNBC) must make dynamic changes in cell shape. The shape of all eukaryotic cells is regulated by Rho Guanine Nucleotide Exchange Factors (RhoGEFs), which activate Rho-family GTPases in response to mechanical and informational cues. In contrast, Rho GTPase-activating proteins (RhoGAPs) inhibit Rho GTPases. However, which RhoGEFs and RhoGAPS couple TNBC cell shape to changes in their environment is very poorly understood. Moreover, whether the activity of particular RhoGEFs and RhoGAPs become dysregulated as cells evolve the ability to metastasise is not clear. Towards the ultimate goal of identifying RhoGEFs and RhoGAPs that are essential for TNBC metastasis, we performed an RNAi screen to isolate RhoGEFs and RhoGAPs that contribute to the morphogenesis of the highly metastatic TNBC cell line LM2, and its less-metastatic parental cell line MDA-MB-231. For ~6 million cells from each cell line, we measured 127 different features following the depletion of 142 genes. Using a linear classifier scheme we also describe the morphological heterogeneity of each gene-depleted population.
Highlights
Background & SummaryThe spread of metastatic cells around the body, and subsequent colonisation of distant sites, is the leading cause of breast cancer patient death[1,2]
Cell shape is regulated by Rho GTPases, which are inactive in a GDP-bound form, but undergo a conformational change upon the binding of GTP, which allows them to activate and recruit downstream effectors[6]
Rho GTP Exchange Factors (RhoGEFs) activate Rho GTPases by inducing the exchange of bound GDP for GTP on Rho GTPases[7]; whereas Rho GTPase-activating proteins (RhoGAPs) inhibit Rho GTPase activity by accelerating GTP hydrolysis[8,9,10]
Summary
Background & SummaryThe spread of metastatic cells around the body, and subsequent colonisation of distant sites, is the leading cause of breast cancer patient death[1,2]. We present both image data, as well as 127 features describing the average cell shape and YAP/TAZ localisation for each well (siRNA) (Fig. 1).
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