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Mechanotransduction and YAP-dependent matrix remodelling is required for the generation and maintenance of cancer-associated fibroblasts

Abstract

To learn more about cancer-associated fibroblasts (CAFs), we have isolated fibroblasts from different stages of breast cancer progression and analysed their function and gene expression. These analyses reveal that activation of the YAP transcription factor is a signature feature of CAFs. YAP function is required for CAFs to promote matrix stiffening, cancer cell invasion and angiogenesis. Remodelling of the ECM and promotion of cancer cell invasion requires the actomyosin cytoskeleton. YAP regulates the expression of several cytoskeletal regulators, including ANLN and DIAPH3, and controls the protein levels of MYL9 (also known as MLC2). Matrix stiffening further enhances YAP activation, thus establishing a feed-forward self-reinforcing loop that helps to maintain the CAF phenotype. Actomyosin contractility and Src function are required for YAP activation by stiff matrices. Further, transient ROCK inhibition is able to disrupt the feed-forward loop, leading to a long-lasting reversion of the CAF phenotype.

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Figure 1: Isolation and characterization of fibroblasts from different disease stages.
Figure 2: YAP is activated in CAFs.
Figure 3: YAP is activated in CAFs in human disease.
Figure 4: YAP is required for CAF function.
Figure 5: Identification of YAP-regulated mRNA and proteins required for CAF function.
Figure 6: Actomyosin- and Src-dependence of YAP activation in CAFs.
Figure 7: Model outlining the role of YAP in the generation and maintenance of CAFs.

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Acknowledgements

F.C., N.E., S.H., R.P.J., S.I.C., K.H. and E.S. are financially supported by Cancer Research UK. A.G-G. was financially supported by a Royal Society Newton Fellowship, E.M. is in receipt of a Dorothy Hodgkins Postgraduate Award (DHPA) from the Engineering and Physical Sciences Research Council. G.C. is in receipt of a Royal Society University Research Fellowship. We thank N. Tapon, B. Thompson and laboratory members for help and advice throughout this work.

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F.C. carried out all the experiments except those noted otherwise. N.E. performed all the quantitative real-time PCR analyses and generated data for Fig. 7e. E.S. generated data for Figs 3, 6g and 7b. E.M. and G.C. performed all the AFM analyses. A.G-G. and S.H. isolated and immortalized several breast and human CAFs. R.P.J. wrote the script for organotypic invasion quantification and helped analyse data for Supplementary Fig. S2c,e. S.I.C., K.H. and P.W. provided clinical material. F.C. and E.S. conceived the study and wrote the manuscript.

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Correspondence to Erik Sahai.

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Calvo, F., Ege, N., Grande-Garcia, A. et al. Mechanotransduction and YAP-dependent matrix remodelling is required for the generation and maintenance of cancer-associated fibroblasts. Nat Cell Biol 15, 637–646 (2013). https://doi.org/10.1038/ncb2756

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