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A gain-of-functional screen identifies the Hippo pathway as a central mediator of receptor tyrosine kinases during tumorigenesis

Oncogene volume 39, pages 334–355 (2020)Cite this article

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Abstract

The Hippo pathway has emerged as a key signaling pathway that regulates various biological functions. Dysregulation of the Hippo pathway has been implicated in a broad range of human cancer types. While a number of stimuli affecting the Hippo pathway have been reported, its upstream kinase and extracellular regulators remain largely unknown. Here we performed the first comprehensive gain-of-functional screen for receptor tyrosine kinases (RTKs) regulating the Hippo pathway using an RTK overexpression library and a Hippo signaling activity biosensor. Surprisingly, we found that the majority of RTKs could regulate the Hippo signaling activity. We further characterized several of these novel relationships [TAM family members (TYRO3, AXL, METRK), RET, and FGFR family members (FGFR1 and FGFR2)] and found that the Hippo effectors YAP/TAZ are central mediators of the tumorigenic phenotypes (e.g., increased cell proliferation, transformation, increased cell motility, and angiogenesis) induced by these RTKs and their extracellular ligands (Gas6, GDNF, and FGF) through either PI3K or MAPK signaling pathway. Significantly, we identify FGFR, RET, and MERTK as the first RTKs that can directly interact with and phosphorylate YAP/TAZ at multiple tyrosine residues independent of upstream Hippo signaling, thereby activating their functions in tumorigenesis. In conclusion, we have identified several novel kinases and extracellular stimuli regulating the Hippo pathway. Our findings also highlight the pivotal role of the Hippo pathway in mediating Gas6/GDNF/FGF-TAM/RET/FGFR-MAPK/PI3K signaling during tumorigenesis and provide a compelling rationale for targeting YAP/TAZ in RTK-driven cancers.

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Acknowledgements

This work was supported by grants from Canadian Institute of Health Research (CIHR#119325, 148629), Canadian Breast Cancer Foundation to XY and CIHR (CIHR#142303) to LMM. TA is supported by the Vanier Canada Graduate Scholarship and Ontario International Graduate Scholarship. HJJvR was supported by a Queen Elizabeth II Graduate Scholarship in Science and Technology. SMM was supported by a Master’s CIHR award. SMM, TM, PK, and TA are supported by studentships from the Terry Fox Research Institute Training Program in Transdisciplinary Cancer Research. We thank Dr David Lillicrap and Dr Paula James for providing the HUVEC, Telo-HEC, and BOEC cells; Dr Camargo for STBS-luc reporter plasmid; Mina Ghahremani for help in making figures; Dr Abdi Ghafari for Aortic ring angiogenesis assay; Elham Ghourbanpour and Dr Lillicrap for helping in Nucleofection transfection; Dr Guan for providing us with HEK293A-LATS knockout cells, and Carrie Wei for help in analyzing motility assay; and Dr Peter Greer for reading the manuscript.

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  1. Department of Pathology and Molecular Medicine, Queen’s University, Kingston, ON, K7L 3N6, Canada

    Taha Azad, Kazem Nouri, Helena J. Janse van Rensburg, Sarah M. Maritan, Liqing Wu, Yawei Hao, Tess Montminy, Jihang Yu, Prem Khanal, Lois M. Mulligan & Xiaolong Yang

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  1. Taha Azad
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TA, LMM and XY designed the study and individual experiments. TA, KN, HJJvR, SMM, LW, YH, JY and PK performed experiments with supervision from XY and LMM. XY and LMM provided resources/equipment and secured funding. TA and XY wrote the manuscript.

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Correspondence to Xiaolong Yang.

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Azad, T., Nouri, K., Janse van Rensburg, H.J. et al. A gain-of-functional screen identifies the Hippo pathway as a central mediator of receptor tyrosine kinases during tumorigenesis. Oncogene 39, 334–355 (2020). https://doi.org/10.1038/s41388-019-0988-y

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