title: α5β1 integrin recycling promotes Arp2/3-independent cancer cell invasion via the formin FHOD3.
abstract: Invasive migration in 3D extracellular matrix (ECM) is crucial to cancer metastasis, yet little is known of the molecular mechanisms that drive reorganization of the cytoskeleton as cancer cells disseminate in vivo. 2D Rac-driven lamellipodial migration is well understood, but how these features apply to 3D migration is not clear. We find that lamellipodia-like protrusions and retrograde actin flow are indeed observed in cells moving in 3D ECM. However, Rab-coupling protein (RCP)-driven endocytic recycling of α5β1 integrin enhances invasive migration of cancer cells into fibronectin-rich 3D ECM, driven by RhoA and filopodial spike-based protrusions, not lamellipodia. Furthermore, we show that actin spike protrusions are Arp2/3-independent. Dynamic actin spike assembly in cells invading in vitro and in vivo is regulated by Formin homology-2 domain containing 3 (FHOD3), which is activated by RhoA/ROCK, establishing a novel mechanism through which the RCP-α5β1 pathway reprograms the actin cytoskeleton to promote invasive migration and local invasion in vivo.
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Nikki R
authors.initials: NR
authors.lastname: Paul
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Allen
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Chapman
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Maria
authors.initials: M
authors.lastname: Morlan-Mairal
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Egor
authors.initials: E
authors.lastname: Zindy
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Jacquemet
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Laura
authors.initials: L
authors.lastname: Fernandez del Ama
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Nermina
authors.initials: N
authors.lastname: Ferizovic
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: David M
authors.initials: DM
authors.lastname: Green
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, England, UK.
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Howe
authors.affiliation: Randall Division of Cell and Molecular Biophysics, Cardiovascular Division, BHF Research Excellence Centre, King's College London, London SE1 1UL, England, UK.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Ehler
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Adam
authors.initials: A
authors.lastname: Hurlstone
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, England, UK patrick.caswell@manchester.ac.uk.
authors.forename: Patrick T
authors.initials: PT
authors.lastname: Caswell
mesh_terms: D051377:Actin-Related Protein 2 / Q000235:genetics / Q000378:metabolism*; D051378:Actin-Related Protein 3 / Q000235:genetics / Q000378:metabolism*; D000199:Actins / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement; D005260:Female; D000080865:Formins; D006801:Humans; D039081:Integrin alpha5beta1 / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D010766:Phosphorylation; D021381:Protein Transport; D011554:Pseudopodia / Q000378:metabolism* / Q000473:pathology; D034622:RNA Interference; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D015027:Zebrafish; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: C495926:ACTR2 protein, human; C495928:ACTR3 protein, human; D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; C485994:FHOD3 protein, human; D000080865:Formins; D039081:Integrin alpha5beta1; D008565:Membrane Proteins; D008840:Microfilament Proteins; C488966:RAB11FIP1 protein, human; D054460:rho-Associated Kinases