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YAP-dependent induction of amphiregulin identifies a non-cell-autonomous component of the Hippo pathway

Abstract

The Hippo signalling pathway regulates cellular proliferation and survival, thus has profound effects on normal cell fate and tumorigenesis1,2,3. The pivotal effector of this pathway is YAP (yes-associated protein), a transcriptional co-activator amplified in mouse and human cancers, where it promotes epithelial to mesenchymal transition (EMT) and malignant transformation4,5,6,7,8,9,10. So far, studies of YAP target genes have focused on cell-autonomous mediators; here we show that YAP-expressing MCF10A breast epithelial cells enhance the proliferation of neighbouring untransfected cells, implicating a non-cell-autonomous mechanism. We identify the gene for the epidermal growth factor receptor (EGFR) ligand amphiregulin (AREG) as a transcriptional target of YAP, whose induction contributes to YAP-mediated cell proliferation and migration, but not EMT. Knockdown of AREG or addition of an EGFR kinase inhibitor abrogates the proliferative effects of YAP expression. Suppression of the negative YAP regulators LATS1 and 2 (large tumour suppressor 1 and 2) is sufficient to induce AREG expression, consistent with physiological regulation of AREG by the Hippo pathway. Genetic interaction between the Drosophila YAP orthologue Yorkie and Egfr signalling components supports the link between these two highly conserved signalling pathways. Thus, YAP-dependent secretion of AREG indicates that activation of EGFR signalling is an important non-cell-autonomous effector of the Hippo pathway, which has implications for the regulation of both physiological and malignant cell proliferation.

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Figure 1: A YAP-induced secreted factor enhances EGF-independent growth of MCF10A cells.
Figure 2: Amphiregulin (AREG) mediates YAP-induced, EGF-independent growth.
Figure 3: Regulation of AREG by the Hippo pathway.
Figure 4: Requirement of AREG for YAP-mediated cell migration and 3D acini formation.
Figure 5: Evolutionary conservation of interactions between EGFR and Hippo pathways in Drosophila.

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Acknowledgements

We thank D. Pan, M. Frolov, G. Halder and Z. Lai for providing Drosophila stocks. We appreciate discussions with J. Wells, F. Yang and T. Zhang and thank B. Fowle for assistance with electron microscopy. This work was supported by the National Institutes of Health (NIH) grant R01 95281, the Doris Duke Foundation Distinguished Clinical Investigator Award, the National Foundation for Cancer Research grant and the Howard Hughes Medical Institute (to D.A.H.); NIH grants CA080111 and CA089393 and the Breast Cancer Research Foundation (to J.S.B.); NIH T32 training grants CA09361-27 (to J.Z.) and CA09361 (to M.O.); NIH grants F32 CA117737 (to G.A.S.); NIH grants GM81607 and GM053203 and the Saltonstall Foundation (to N.J.D.); and the Tosteson postdoctoral Fellowship (to J.Y.J.).

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J.Z., J.J., M.O., J.S.B., N.J.D. and D.A.H. designed the research; J.Z., J.J., M.Y., M.O., G.A.S. and R.W. performed experiments; J.Z., J.J., M.O., J.S.B., N.J.D. and D.A.H. analysed data; and J.Z., J.J., J.S.B., N.J.D. and D.A.H. wrote the paper.

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Correspondence to Daniel A. Haber.

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Zhang, J., Ji, JY., Yu, M. et al. YAP-dependent induction of amphiregulin identifies a non-cell-autonomous component of the Hippo pathway. Nat Cell Biol 11, 1444–1450 (2009). https://doi.org/10.1038/ncb1993

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