WIP is necessary for matrix invasion by breast cancer cells.

Actin filament assembly and reorganisation during cell migration and invasion into extracellular matrices is a well-documented phenomenon. Among actin-binding proteins regulating its polymerisation, the members of the WASP (Wiskott Aldrich Syndrome Protein) family are generally thought to play the most significant role in supporting cell invasiveness. In situ, cytosolic N-WASP (neural WASP) is associated with a partner protein termed WIP (WASP Interacting Protein) that is bound to the N-terminal domain of N-WASP. Despite much effort, rather little is known about the role of WIP in regulating N-WASP and consequent actin-filament assembly. Even less is known about the function of WIP within the specialised cell adhesion and attachment structures known as podosomes and invadopodia. In particular, whilst the interaction of WIP with known participants in the development and maturation of invadopodia such as N-WASP, the Arp2/3 complex and cortactin has been described, little is known concerning the direct contribution of WIP to invadopodia and its potential role as a regulator of cancer cell invasion. In this report, we use 2D and 3D culture systems to describe the role played by WIP in modulating the morphology and invasiveness of metastatic breast cancer cells in vitro, as well as its effect on the process of mesenchymal-epithelial transition (MET) seen in these cells. We demonstrate that WIP is necessary for invadopodium formation and matrix degradation by basal breast cancer cells, but not sufficient to induce invasiveness in luminal cells.

García E ，Machesky LM ，Jones GE ，Antón IM ... -  《-》

WIP and WICH/WIRE co-ordinately control invadopodium formation and maturation in human breast cancer cell invasion.

García E ，Ragazzini C ，Yu X ，Cuesta-García E ，Bernardino de la Serna J ，Zech T ，Sarrió D ，Machesky LM ，Antón IM ... -  《Scientific Reports》

Invadopodia proteins, cortactin, N-WASP and WIP differentially promote local invasiveness in ameloblastoma.

Cell migration and invasion through interstitial tissues are dependent upon several specialized characteristics of the migratory cell notably generation of proteolytic membranous protrusions or invadopodia. Ameloblastoma is a benign odontogenic epithelial neoplasm with a locally infiltrative behaviour. Cortactin and MMT1-MMP are two invadopodia proteins implicated in its local invasiveness. Other invadopodia regulators, namely N-WASP, WIP and Src kinase remain unclarified. This study addresses their roles in ameloblastoma. Eighty-seven paraffin-embedded ameloblastoma cases (20 unicystic, 47 solid/multicystic, 3 desmoplastic and 17 recurrent) were subjected to immunohistochemistry for expression of cortactin, N-WASP, WIP, Src kinase and F-actin, and findings correlated with clinicopathological parameters. Invadopodia proteins (except Src kinase) and F-actin were widely detected in ameloblastoma (cortactin: n = 73/87, 83.9%; N-WASP: n = 59/87; 67.8%; WIP: n = 77/87; 88.5%; and F-actin: n = 87/87, 100%). Protein localization was mainly cytoplasmic and/or membranous, and occasionally nuclear for F-actin. Cortactin, which functions as an actin-scaffolding protein, demonstrated significantly higher expression levels within ameloblastoma tumoral epithelium than in stroma (P < 0.05). N-WASP, which coordinates actin polymerization and invadopodia-mediated extracellular matrix degradation, was overexpressed in the solid/multicystic subtype (P < 0.05). WIP, an upstream regulator of N-WASP, and F-actin were significantly upregulated along the tumour invasive front compared to tumour centres (P < 0.05). Except for males with cortactin overexpression, other clinical parameters (age, ethnicity and anatomical site) showed no significant correlations. Present results suggest that local invasiveness of ameloblastoma is dependent upon the migratory potential of its tumour cells as defined by their distribution of cortactin, N-WASP and WIP in correlation with F-actin cytoskeletal dynamics.

Siar CH ，Rahman ZA ，Tsujigiwa H ，Mohamed Om Alblazi K ，Nagatsuka H ，Ng KH ... -  《-》

WIP: WASP-interacting proteins at invadopodia and podosomes.

Regulated cell invasion resulting from migratory and matrix-degrading events is an essential step in physiological processes such as the inflammatory response and tissue repair. Cell invasion is also thought to be a critical parameter in pathological conditions such as cancer metastasis. The migration of normal and cancer cells is largely driven by the actin cytoskeleton, which controls cell shape, adhesion and contractility. Podosomes and invadopodia are actin-rich protrusions that drive invasion in normal and cancer cells. These structures protrude from the basal region of the cell facing the extracellular matrix, where they adhere to and degrade the matrix, thus facilitating invasive migration. WASP (Wiskott-Aldrich syndrome protein) and WIP (WASP-interacting protein) localise to the actin rich core of podosomes and play a critical role in their formation. More recently, studies performed on microarray data sets from cancer patients of several tumour categories show a strong correlation between reduced WIP expression and improved prognosis. In this article, we identify endogenous WIP at the distal tips of cancer cell invasive protrusions and we summarise recent advances in the study of the roles of WIP- and WASP-protein families during migration and invasion of normal and cancer cells related to podosome and invadopodium generation.

García E ，Jones GE ，Machesky LM ，Antón IM ... -  《-》

Intersectin adaptor proteins are associated with actin-regulating protein WIP in invadopodia.

Invasive cancer cells form actin-rich membrane protrusions called invadopodia that degrade extracellular matrix and facilitate cell invasion and metastasis. WIP (WASP-interacting protein) together with N-WASP (neural Wiskott-Aldrich syndrome protein) are localized in invadopodia and play a crucial role in their formation. Here we show that WIP interacts with endocytic adaptor proteins of the intersectin (ITSN) family, ITSN1 and ITSN2. The interaction is mediated by the SH3 domains of ITSNs and the middle part of the WIP proline-rich motifs. We have also demonstrated that ITSN1, WIP and N-WASP can form a complex in cells. Endogenous ITSN1 and ITSN2 are located in invasive protrusions of MDA-MB-231 breast cancer cell line. Moreover, data from immunofluorescent analysis revealed co-localization of ITSN1 and WIP at sites of invadopodia formation and in clathrin-coated pits. Together, these findings provide insights into the molecular mechanisms of invadopodia formation and identify ITSNs as scaffold proteins involved in this process.

Gryaznova T ，Kropyvko S ，Burdyniuk M ，Gubar O ，Kryklyva V ，Tsyba L ，Rynditch A ... -  《-》