title: Inhibitory cortactin nanobodies delineate the role of NTA- and SH3-domain-specific functions during invadopodium formation and cancer cell invasion.
abstract: Cancer cells exploit different strategies to escape from the primary tumor, gain access to the circulation, disseminate throughout the body, and form metastases, the leading cause of death by cancer. Invadopodia, proteolytically active plasma membrane extensions, are essential in this escape mechanism. Cortactin is involved in every phase of invadopodia formation, and its overexpression is associated with increased invadopodia formation, extracellular matrix degradation, and cancer cell invasion. To analyze endogenous cortactin domain function in these processes, we characterized the effects of nanobodies that are specific for the N-terminal acidic domain of cortactin and expected to target small epitopes within this domain. These nanobodies inhibit cortactin-mediated actin-related protein (Arp)2/3 activation, and, after their intracellular expression in cancer cells, decrease invadopodia formation, extracellular matrix degradation, and cancer cell invasion. In addition, one of the nanobodies affects Arp2/3 interaction and invadopodium stability, and a nanobody targeting the Src homology 3 domain of cortactin enabled comparison of 2 functional regions in invadopodium formation or stability. Given their common and distinct effects, we validate cortactin nanobodies as an instrument to selectively block and study distinct domains within a protein with unprecedented precision, aiding rational future generation of protein domain-selective therapeutic compounds.-Bertier, L., Boucherie, C., Zwaenepoel, O., Vanloo, B., Van Troys, M., Van Audenhove, I., Gettemans, J. Inhibitory cortactin nanobodies delineate the role of NTA- and SH3-domain-specific functions during invadopodium formation and cancer cell invasion.
authors.affiliation: Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University-Campus Rommelaere, Ghent, Belgium.
authors.forename: Laurence
authors.initials: L
authors.lastname: Bertier
authors.affiliation: Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University-Campus Rommelaere, Ghent, Belgium.
authors.forename: Ciska
authors.initials: C
authors.lastname: Boucherie
authors.affiliation: Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University-Campus Rommelaere, Ghent, Belgium.
authors.forename: Olivier
authors.initials: O
authors.lastname: Zwaenepoel
authors.affiliation: Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University-Campus Rommelaere, Ghent, Belgium.
authors.forename: Berlinda
authors.initials: B
authors.lastname: Vanloo
authors.affiliation: Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University-Campus Rommelaere, Ghent, Belgium.
authors.forename: Marleen
authors.initials: M
authors.lastname: Van Troys
authors.affiliation: Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University-Campus Rommelaere, Ghent, Belgium.
authors.forename: Isabel
authors.initials: I
authors.lastname: Van Audenhove
authors.affiliation: Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University-Campus Rommelaere, Ghent, Belgium jan.gettemans@ugent.be.
authors.forename: Jan
authors.initials: J
authors.lastname: Gettemans
mesh_terms: D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D051356:Cortactin / Q000737:chemistry* / Q000378:metabolism; D000939:Epitopes; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D009361:Neoplasm Invasiveness; D000069261:Podosomes / Q000502:physiology*; D000072417:Protein Domains; D061905:Single-Domain Antibodies / Q000502:physiology*
chemical_list: C543733:CK-0944666; D051356:Cortactin; D000939:Epitopes; D007211:Indoles; D061905:Single-Domain Antibodies
keywords: Arp2/3 complex; actin; intrabody; matrix metalloprotease; single domain antibody