title: Cellular adhesome screen identifies critical modulators of focal adhesion dynamics, cellular traction forces and cell migration behaviour.
abstract: Cancer cells migrate from the primary tumour into surrounding tissue in order to form metastasis. Cell migration is a highly complex process, which requires continuous remodelling and re-organization of the cytoskeleton and cell-matrix adhesions. Here, we aimed to identify genes controlling aspects of tumour cell migration, including the dynamic organization of cell-matrix adhesions and cellular traction forces. In a siRNA screen targeting most cell adhesion-related genes we identified 200+ genes that regulate size and/or dynamics of cell-matrix adhesions in MCF7 breast cancer cells. In a subsequent secondary screen, the 64 most effective genes were evaluated for growth factor-induced cell migration and validated by tertiary RNAi pool deconvolution experiments. Four validated hits showed significantly enlarged adhesions accompanied by reduced cell migration upon siRNA-mediated knockdown. Furthermore, loss of PPP1R12B, HIPK3 or RAC2 caused cells to exert higher traction forces, as determined by traction force microscopy with elastomeric micropillar post arrays, and led to considerably reduced force turnover. Altogether, we identified genes that co-regulate cell-matrix adhesion dynamics and traction force turnover, thereby modulating overall motility behaviour.
authors.affiliation: Leiden Academic Centre for Drug Research, Division of Toxicology, Leiden University, The Netherlands.
authors.forename: Michiel
authors.initials: M
authors.lastname: Fokkelman
authors.affiliation: Leiden Academic Centre for Drug Research, Division of Toxicology, Leiden University, The Netherlands.
authors.forename: Hayri E
authors.initials: HE
authors.lastname: Balcıoğlu
authors.affiliation: Leiden Academic Centre for Drug Research, Division of Toxicology, Leiden University, The Netherlands.
authors.forename: Janna E
authors.initials: JE
authors.lastname: Klip
authors.affiliation: Imaging &Bioinformatics, Leiden Institute of Advanced Computer Science, Leiden University, The Netherlands.
authors.forename: Kuan
authors.initials: K
authors.lastname: Yan
authors.affiliation: Imaging &Bioinformatics, Leiden Institute of Advanced Computer Science, Leiden University, The Netherlands.
authors.forename: Fons J
authors.initials: FJ
authors.lastname: Verbeek
authors.affiliation: Leiden Academic Centre for Drug Research, Division of Toxicology, Leiden University, The Netherlands.
authors.forename: Erik H J
authors.initials: EH
authors.lastname: Danen
authors.affiliation: Leiden Academic Centre for Drug Research, Division of Toxicology, Leiden University, The Netherlands.
authors.forename: Bob
authors.initials: B
authors.lastname: van de Water
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D002465:Cell Movement; D005109:Extracellular Matrix / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D005260:Female; D022001:Focal Adhesions / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D061986:MCF-7 Cells; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D009363:Neoplasm Proteins