title: NEDD9/Arf6-dependent endocytic trafficking of matrix metalloproteinase 14: a novel mechanism for blocking mesenchymal cell invasion and metastasis of breast cancer.
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.
authors.affiliation: Mary Babb Randolph Cancer Center, West Virginia University School of Medicine, Morgantown, WV, USA.
authors.forename: Y V
authors.initials: YV
authors.lastname: Loskutov
authors.affiliation: Department of Biochemistry, West Virginia University School of Medicine, Morgantown, WV, USA.
authors.forename: P Y
authors.initials: PY
authors.lastname: Kozyulina
authors.affiliation: Mary Babb Randolph Cancer Center, West Virginia University School of Medicine, Morgantown, WV, USA.
authors.forename: V K
authors.initials: VK
authors.lastname: Kozyreva
authors.affiliation: Mary Babb Randolph Cancer Center, West Virginia University School of Medicine, Morgantown, WV, USA.
authors.forename: R J
authors.initials: RJ
authors.lastname: Ice
authors.affiliation: Department of Biochemistry, West Virginia University School of Medicine, Morgantown, WV, USA.
authors.forename: B C
authors.initials: BC
authors.lastname: Jones
authors.affiliation: Department of Biochemistry, West Virginia University School of Medicine, Morgantown, WV, USA.
authors.forename: T J
authors.initials: TJ
authors.lastname: Roston
authors.affiliation: Department of Pathology, West Virginia University School of Medicine, Morgantown, WV, USA.
authors.forename: M B
authors.initials: MB
authors.lastname: Smolkin
authors.affiliation: 1] Mary Babb Randolph Cancer Center, West Virginia University School of Medicine, Morgantown, WV, USA [2] Department of Biochemistry, West Virginia University School of Medicine, Morgantown, WV, USA.
authors.forename: A V
authors.initials: AV
authors.lastname: Ivanov
authors.affiliation: 1] Mary Babb Randolph Cancer Center, West Virginia University School of Medicine, Morgantown, WV, USA [2] Department of Neurobiology and Anatomy, West Virginia University School of Medicine, Morgantown, WV, USA.
authors.forename: R B
authors.initials: RB
authors.lastname: Wysolmerski
authors.affiliation: 1] Mary Babb Randolph Cancer Center, West Virginia University School of Medicine, Morgantown, WV, USA [2] Department of Biochemistry, West Virginia University School of Medicine, Morgantown, WV, USA.
authors.forename: E N
authors.initials: EN
authors.lastname: Pugacheva
mesh_terms: D000089982:ADP-Ribosylation Factor 6; D020727:ADP-Ribosylation Factors / Q000378:metabolism*; D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement; D011992:Endosomes / Q000378:metabolism*; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism*; D008648:Mesoderm / Q000166:cytology / Q000473:pathology; D051379:Mice; D009362:Neoplasm Metastasis; D009368:Neoplasm Transplantation; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010750:Phosphoproteins / Q000378:metabolism*; D015398:Signal Transduction; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000378:metabolism*
chemical_list: D000089982:ADP-Ribosylation Factor 6; D048868:Adaptor Proteins, Signal Transducing; C099942:NEDD9 protein, human; D016376:Oligonucleotides, Antisense; D010750:Phosphoproteins; C560823:TIMP2 protein, human; D019716:Tissue Inhibitor of Metalloproteinase-2; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14; D020727:ADP-Ribosylation Factors; C000715348:ARF6 protein, human; C000715727:Arf6 protein, mouse