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Neratinib inhibits Hippo/YAP signaling, reduces mutant K-RAS expression, and kills pancreatic and blood cancer cells

Oncogene volume 38, pages 5890–5904 (2019)Cite this article

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Abstract

Prior studies demonstrated that the irreversible ERBB1/2/4 inhibitor neratinib caused plasma membrane-associated mutant K-RAS to localize in intracellular vesicles, concomitant with its degradation. Herein, we discovered that neratinib interacted with the chemically distinct irreversible ERBB1/2/4 inhibitor afatinib to reduce expression of ERBB1, ERBB2, K-RAS and N-RAS; this was associated with greater-than-additive cell killing of pancreatic tumor cells. Knock down of Beclin1, ATG16L1, Rubicon or cathepsin B significantly lowered the ability of neratinib to reduce ERBB1 and K-RAS expression, and to cause tumor cell death. Knock down of ATM-AMPK suppressed vesicle formation and knock down of cathepsin B-AIF significantly reduced neratinib lethality. PKG phosphorylates K-RAS and HMG CoA reductase inhibitors reduce K-RAS farnesylation both of which remove K-RAS from the plasma membrane, abolishing its activity. Neratinib interacted with the PKG activator sildenafil and the HMG CoA reductase inhibitor atorvastatin to further reduce K-RAS expression, and to further enhance cell killing. Neratinib is also a Ste20 kinase family inhibitor and in carcinoma cells, and hematopoietic cancer cells lacking ERBB1/2/4, it reduced K-RAS expression and the phosphorylation of MST1/3/4/Ezrin by ~ 30%. Neratinib increased LATS1 phosphorylation as well as that of YAP and TAZ also by ~ 30%, caused the majority of YAP to translocate into the cytosol and reduced YAP/TAZ protein levels. Neratinib lethality was enhanced by knock down of YAP. Neratinib, in a Rubicon-dependent fashion, reduced PAK1 phosphorylation and that of its substrate Merlin. Our data demonstrate that neratinib coordinately suppresses both mutant K-RAS and YAP function to kill pancreatic tumor cells.

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Acknowledgements

PD acknowledges funding by the Commonwealth Health Research Board (CHRB) of Virginia. JFH acknowledges funding by CPRIT RP170233.

Funding

Support for the present study was funded from philanthropic funding from Massey Cancer Center and the Universal Inc. Chair in Signal Transduction Research.

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Authors and Affiliations

  1. Departments of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA, 23298-0035, USA

    Paul Dent, Laurence Booth & Jane L. Roberts

  2. Department of Integrative Biology and Pharmacology, University of Texas Health Science Center, Houston, TX, 77030, USA

    Junchen Liu & John F. Hancock

  3. Departments of Medicine, Virginia Commonwealth University, Richmond, VA, 23298-0035, USA

    Andrew Poklepovic

  4. Puma Biotechnology Inc., 1880 Wilshire Blvd, Los Angeles, CA, 90024, USA

    Alshad S. Lalani

  5. Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY, 11724, USA

    David Tuveson

  6. Inflammation & Autoimmunity Group, NIEHS, Research Triangle Park, NC, 27709, USA

    Jennifer Martinez

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Dent, P., Booth, L., Roberts, J.L. et al. Neratinib inhibits Hippo/YAP signaling, reduces mutant K-RAS expression, and kills pancreatic and blood cancer cells. Oncogene 38, 5890–5904 (2019). https://doi.org/10.1038/s41388-019-0849-8

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