title: Regulated delivery of molecular cargo to invasive tumour-derived microvesicles.
abstract: Cells release multiple, distinct forms of extracellular vesicles including structures known as microvesicles, which are known to alter the extracellular environment. Despite growing understanding of microvesicle biogenesis, function and contents, mechanisms regulating cargo delivery and enrichment remain largely unknown. Here we demonstrate that in amoeboid-like invasive tumour cell lines, the v-SNARE, VAMP3, regulates delivery of microvesicle cargo such as the membrane-type 1 matrix metalloprotease (MT1-MMP) to shedding microvesicles. MT1-MMP delivery to nascent microvesicles depends on the association of VAMP3 with the tetraspanin CD9 and facilitates the maintenance of amoeboid cell invasion. VAMP3-shRNA expression depletes shed vesicles of MT1-MMP and decreases cell invasiveness when embedded in cross-linked collagen matrices. Finally, we describe functionally similar microvesicles isolated from bodily fluids of ovarian cancer patients. Together these studies demonstrate the importance of microvesicle cargo sorting in matrix degradation and disease progression.
authors.affiliation: Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors.forename: James W
authors.initials: JW
authors.lastname: Clancy
authors.affiliation: Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors.forename: Alanna
authors.initials: A
authors.lastname: Sedgwick
authors.affiliation: Institut Curie, Centre de Recherche, Paris F-75248, France.
authors.forename: Carine
authors.initials: C
authors.lastname: Rosse
authors.affiliation: Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors.forename: Vandhana
authors.initials: V
authors.lastname: Muralidharan-Chari
authors.affiliation: Institut Curie, Centre de Recherche, Paris F-75248, France.
authors.forename: Graca
authors.initials: G
authors.lastname: Raposo
authors.affiliation: Northern Indiana Cancer Consortium, Michiana Hematology Oncology, Mishawaka, Indiana 46545, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Method
authors.affiliation: Institut Curie, Centre de Recherche, Paris F-75248, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Chavrier
authors.affiliation: Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors.forename: Crislyn
authors.initials: C
authors.lastname: D'Souza-Schorey
mesh_terms: D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D005456:Fluorescent Dyes; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D006801:Humans; D008545:Melanoma; D009361:Neoplasm Invasiveness; D009369:Neoplasms / Q000098:blood supply*; D009389:Neovascularization, Pathologic / Q000378:metabolism*; D021381:Protein Transport; D051040:Vesicle-Associated Membrane Protein 3 / Q000235:genetics / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; C494946:VAMP3 protein, human; D051040:Vesicle-Associated Membrane Protein 3