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NEDD9/Arf6-dependent endocytic trafficking of matrix metalloproteinase 14: a novel mechanism for blocking mesenchymal cell invasion and metastasis of breast cancer

Oncogene volume 34, pages 3662–3675 (2015)Cite this article

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Abstract

NEDD9 is an established marker of invasive and metastatic cancers. NEDD9 downregulation has been shown to dramatically reduce cell invasion and metastasis in multiple tumors. The mechanisms by which NEDD9 regulates invasion are largely unknown. In the current study, we have found that NEDD9 is required for matrix metalloproteinase 14 (MMP14) enzymatic recovery/recycling through the late endosomes to enable disengagement of tissue inhibitor of matrix metalloproteinase 2 (TIMP2) and tumor invasion. Depletion of NEDD9 decreases targeting of the MMP14/TIMP2 complex to late endosomes and increases trafficking of MMP14 from early/sorting endosomes back to the surface in a small GTPase ADP ribosylation factor-6 (Arf6)-dependent manner. NEDD9 directly binds to Arf6-GTPase-activating protein, ARAP3 and Arf6-effector GGA3, thereby facilitating the Arf6 inactivation required for MMP14/TIMP2 targeting to late endosomes. Re-expression of NEDD9 or a decrease in Arf6 activity is sufficient to restore MMP14 activity and the invasive properties of tumor cells. Importantly, NEDD9 inhibition by Vivo-Morpholinos, an antisense therapy, decreases primary tumor growth and metastasis in xenograft models of breast cancer. Collectively, our findings uncover a novel mechanism to control tumor-cell dissemination through NEDD9/Arf6-dependent regulation of MMP14/TIMP2 trafficking, and validate NEDD9 as a clinically relevant therapeutic target to treat metastatic cancer.

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Acknowledgements

We thank Mrs Yoho and Drs Schaller and Weed for critically reading the manuscript and providing administrative support during manuscript preparation. We also thank the West Virginia University Microscope Imaging Facility, supported by the Mary Babb Randolph Cancer Center and NIH grants P20 RR016440, P30 RR032138/GM103488 and P20 RR016477. The WVU Flow Cytometry Core Facility was supported by NIH grants P30 GM103488, P30 RR032138 and RCP1101809. We also thank Mr Farrugia for his assistance in the analysis of the microarray data. This work was supported by grants from the NIH-NCI (CA148671 to ENP), the Susan G Komen For the Cure Foundation (KG100539 to ENP) and in part by NIH/NCRR (5 P20 RR016440-09).

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Authors and Affiliations

  1. Mary Babb Randolph Cancer Center, West Virginia University School of Medicine, Morgantown, WV, USA

    Y V Loskutov, V K Kozyreva, R J Ice, A V Ivanov, R B Wysolmerski & E N Pugacheva

  2. Department of Biochemistry, West Virginia University School of Medicine, Morgantown, WV, USA

    P Y Kozyulina, B C Jones, T J Roston, A V Ivanov & E N Pugacheva

  3. Department of Pathology, West Virginia University School of Medicine, Morgantown, WV, USA

    M B Smolkin

  4. Department of Neurobiology and Anatomy, West Virginia University School of Medicine, Morgantown, WV, USA

    R B Wysolmerski

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Loskutov, Y., Kozyulina, P., Kozyreva, V. et al. NEDD9/Arf6-dependent endocytic trafficking of matrix metalloproteinase 14: a novel mechanism for blocking mesenchymal cell invasion and metastasis of breast cancer. Oncogene 34, 3662–3675 (2015). https://doi.org/10.1038/onc.2014.297

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