Signalling platforms in cancer: molecular and functional characterisation and translational strategies.
The main interest of our lab is to investigate cell signalling originating from cell adhesion and growth factor receptor in the control of cell transformation and cancer cell progression. In our lab we have contributed to define that integrin-dependent adhesion and growth factor/cytokine stimulation co-ordinately regulate convergent signalling pathways that control cell growth, migration and invasion in normal and transformed cells. Our current research is focused on the following points:
• the cross-talk between beta1 integrin and the EGF receptor
• the role of the adaptor protein p130Cas in development and tumorigenesis
• the role of the adaptor protein p140Cap in development and tumorigenesis.
Description
It is well known that when cells become transformed and malignant, they acquire anchorage-independent growth and invasive properties. It is also well established that the signalling originated from the integrin family of cell-matrix receptors is central to the conversion from a normal to a tumour phenotype leading to cell transformation and cancer cell progression (Cabodi and Defilippi, 2006).
In our lab we have contributed to define that integrin-dependent adhesion and growth factor/cytokine stimulation co-ordinately regulate convergent signalling pathways that control cell growth, spreading and migration in normal cells (Moro et al., 1998, 2002; Cabodi, Morello et al. 2009; Defilippi et al., 2005). Therefore understanding the molecular mechanisms by which integrins and growth factors co-operate in the control of cell growth, migration and survival from apoptosis is a fundamental relevant task to the study of cell transformation. Increasing evidences indicate that integrins co-operate with growth factor receptors in the control of cell survival and proliferation, by formation of macromolecular transducing complexes.
In this context we have identified three main signalling platforms into the cells
1) A platform constituted by integrins and the EGF receptor, essential for full activation of EGFR in response to EGF, and for EGF-dependent downstream signalling (Cabodi and Defilippi, 2006; Cabodi, Morello et al., 2009).
2) A platform formed by the adaptor protein p130Cas. Due to the presence of multiple conserved sequence motifs and extensive post-translational modifications, it promotes protein-protein interactions leading to multi-protein complexes (Defilippi et al., 2006). In normal cells, interaction of p130Cas with other proteins modulates cell motility, survival and proliferation. We have already determined the involvement of p130Cas in mammary gland development and tumorigenesis (Cabodi et al., 2006) and we are in the process of analysing the involvement of p130Cas in cell transformation by specific oncogenes.
3) A platform formed by the adaptor protein p140Cap. We recently identified p140Cap as a novel adaptor protein and a downstream effector of cell-matrix and growth factor signalling (Di Stefano et al., 2004; Di Stefano, Damiano et al., 2007). our data show that p140Cap controls Src kinase activity, resulting in decreased cell proliferation, motility and invasion, both in vivo and in vitro, thus regulating tumorigenic properties of breast cancer cells (Damiano, Di Stefano et al., 2010). Over-expression of the p140Cap adaptor might thus constitute a potential tool to revert cancer cells to a more differentiated, less invasive phenotype.
Based on these results we are currently characterising these molecules with molecular and functional approaches, using the following models:
• In vitro cell cultures from breast, lung and colon tumours, over-expressing or silenced for specific components of the different platforms
• Transgenic mice over-expressing p130Cas and p140Cap in the mammary gland
• Knock-out animals for the p140Cap protein
• Selective inhibitors for preclinical studies
• Normal and cancer stem cells derived from the transgenic mice.
Relevant publications:
Laura Damiano, Paola Di Stefano, Maria del Pilar Camacho Leal, Matteo Barba, Fabrizio Mainiero, Sara Cabodi, Luca Tordella, Anna Sapino, Isabella Castellano, Marta Canel, Margaret Frame, Emilia Turco, Paola Defilippi p140Cap Dual Regulation of E-cadherin/EGFR Cross-talk and Ras signalling in Tumour Cell Scatter and Proliferation Oncogene 2010, in press 1: Jaworski J, Kapitein LC, Gouveia SM, Dortland BR, Wulf PS, Grigoriev I, Camera P, Spangler SA, Di Stefano P, Demmers J, Krugers H, Defilippi P, Akhmanova A, Hoogenraad CC. Dynamic microtubules regulate dendritic spine morphology and synaptic plasticity. Neuron. 2009 Jan 15;61(1):85-100. 2: Ito H, Atsuzawa K, Sudo K, Di Stefano P, Iwamoto I, Morishita R, Takei S, Semba R, Defilippi P, Asano T, Usuda N, Nagata K. Characterization of a multidomain adaptor protein, p140Cap, as part of a pre-synaptic complex. J Neurochem. 2008 Oct;107(1):61-72. 3: Di Stefano P, Damiano L, Cabodi S, Aramu S, Tordella L, Praduroux A, Piva R, Cavallo F, Forni G, Silengo L, Tarone G, Turco E, Defilippi P. p140Cap protein suppresses tumour cell properties, regulating Csk and Src kinase activity. EMBO J. 2007 Jun 20;26(12):2843-55. 4: Boeri Erba E, Matthiesen R, Bunkenborg J, Schulze WX, Di Stefano P, Cabodi S, Tarone G, Defilippi P, Jensen ON. Quantitation of multisite EGF receptor phosphorylation using mass spectrometry and a novel normalization approach. J Proteome Res. 2007 Jul;6(7):2768-85. 5: Cabodi S, Tinnirello A, Di Stefano P, Bisarò B, Ambrosino E, Castellano I, Sapino A, Arisio R, Cavallo F, Forni G, Glukhova M, Silengo L, Altruda F, Turco E, Tarone G, Defilippi P. p130Cas as a new regulator of mammary epithelial cell proliferation, survival, and HER2-neu oncogene-dependent breast tumorigenesis. Cancer Res. 2006 May 1;66(9):4672-80. 6: Defilippi P, Di Stefano P, Cabodi S. p130Cas: a versatile scaffold in signaling networks. Trends Cell Biol. 2006 May;16(5):257-63. 7: Ambrogio C, Voena C, Manazza AD, Piva R, Riera L, Barberis L, Costa C, Tarone G, Defilippi P, Hirsch E, Boeri Erba E, Mohammed S, Jensen ON, Palestro G, Inghirami G, Chiarle R. p130Cas mediates the transforming properties of the anaplastic lymphoma kinase. Blood. 2005 Dec 1;106(12):3907-16. 8: Dentelli P, Rosso A, Garbarino G, Calvi C, Lombard E, Di Stefano P, Defilippi P, Pegoraro L, Brizzi MF. The interaction between KDR and interleukin-3 receptor (IL-3R) beta common modulates tumor neovascularization. Oncogene. 2005 Sep 22;24(42):6394-405. 9: Boeri Erba E, Bergatto E, Cabodi S, Silengo L, Tarone G, Defilippi P, Jensen ON. Systematic analysis of the epidermal growth factor receptor by mass spectrometry reveals stimulation-dependent multisite phosphorylation. Mol Cell Proteomics. 2005 Aug;4(8):1107-21. 10: Cherubini A, Hofmann G, Pillozzi S, Guasti L, Crociani O, Cilia E, Di Stefano P, Degani S, Balzi M, Olivotto M, Wanke E, Becchetti A, Defilippi P, Wymore R, Arcangeli A. Human ether-a-go-go-related gene 1 channels are physically linked to beta1 integrins and modulate adhesion-dependent signaling. Mol Biol Cell. 2005 Jun;16(6):2972-83. 11: Cabodi S, Moro L, Baj G, Smeriglio M, Di Stefano P, Gippone S, Surico N, Silengo L, Turco E, Tarone G, Defilippi P. p130Cas interacts with estrogen receptor alpha and modulates non-genomic estrogen signaling in breast cancer cells. J Cell Sci. 2004 Mar 15;117(Pt 8):1603-11. 12: Defilippi P, Vallés AM. The winding road from adhesive receptors to the nucleus. EMBO Rep. 2002 Apr;3(4):312-6. 13: Moro L, Dolce L, Cabodi S, Bergatto E, Boeri Erba E, Smeriglio M, Turco E, Retta SF, Giuffrida MG, Venturino M, Godovac-Zimmermann J, Conti A, Schaefer E, Beguinot L, Tacchetti C, Gaggini P, Silengo L, Tarone G, Defilippi P. Integrin-induced epidermal growth factor (EGF) receptor activation requires c-Src and p130Cas and leads to phosphorylation of specific EGF receptor tyrosines. J Biol Chem. 2002 Mar 15;277(11):9405-14. 14: Moro L, Venturino M, Bozzo C, Silengo L, Altruda F, Beguinot L, Tarone G, Defilippi P. Integrins induce activation of EGF receptor: role in MAP kinase induction and adhesion-dependent cell survival. EMBO J. 1998 Nov 16;17(22):6622-32. |
