Centro Interdipartimentale di Biotecnologie Molecolari "Guido Tarone"
Paola Defilippi - PI
Full Professor of Cellular and Applied Biology, University of Torino, Italy
- Vincenzo Salemme, post-doctoral fellow
- Mario De Gregorio, PhD student
- Francesca Nigrellli, PhD student
- Vittorio Belmonte, Fellow
- Christian Mosoni Fellow
- Master students: Beatrice Bersia, Giulia Dessy, Matteo Poncina, Lucrezia Roetgen, Andrea Scavuzzo, , Noemi Zuccaro
Research activity
Molecular and functional characterization of signaling platforms and translational strategies
Cell matrix adhesion is mainly regulated by the integrin family of extracellular matrix receptors which are crucial for cell life. My group has been involved since the early ‘90 years in studying integrin signaling with special interest in the pathophysiology of integrin- and growth factor- dependent control of cell growth and migration. Over the years, we use cellular and molecular biology techniques, generating new cell and animal models to deeply investigate signaling pathways and their functional relevance. My lab was one of the first to provide evidence of an extensive crosstalk between beta1 integrin and EGF receptor (EGFR) that involves physical interaction among the proteins at the cell surface (Moro et al., EMBO J. 1998 PMID:9822606; Moro et al., JBC 2002 PMID:11756413). Our work also established that Integrin/EGFR cross talk is essential for priming EGFR activity in normal and cancer cells (Morello et al., Oncogene 2011 PMID:21478906).
Defilippi’s group had identified and fully characterized at the genetic and functional level adaptor proteins that build up very upstream molecular platforms in integrin/RTK-dependent signals. Adaptor proteins are generally devoid of any enzymatic or transcriptional activity, and their modular structure with various binding motifs allows the formation of multi-protein signaling complexes. This results in the induction and/or maintenance of signaling pathways with pleiotropic effects on cell motility, cell adhesion, cytoskeleton remodeling, invasion, survival, and proliferation. Among them, the p130Cas (encoded by the BCAR1 gene) and p140Cap (encoded by the SRCIN1 gene) adaptor proteins have been extensively studied in cell and animal models, providing evidence of their role in the control of pathways leading to cytoskeleton organization, cell motility and cell invasion.
Cancer cell Biology
This is particularly relevant in cancer cells, regulating cancer progression (Cabodi et al., Cancer Res 2006 PMID:16651418; Di Stefano et al., EMBO J 2007 PMID:17525734; Damiano et al., Oncogene 2010 PMID:20453886; Cabodi et al., Nat Rev Cancer 2010, PMID:21102636; Bisaro et al., Breast Can Res 2014 PMID:25606587; Grasso et al., Nat Commun 2017 PMID 28300085). The above findings provide evidence for a role of p130Cas as a positive regulator of both proliferation and survival in normal and transformed mammary epithelial cells. In contrast, p140Cap, in the same breast cancer cell models, behaves as an oncosuppressor which opposes and interferes with cancer features (Centonze et al., Front Cell Dev Biol. 2021 PMID: 34708040; Salemme et al., Cell Mol Life Sci 2021 PMID: 33079227). p140Cap behaves as an oncosuppresor also in neuroblastoma (Grasso, Cangelosi et al., Cell Death and Diff 2019 PMID:31488891).
Ongoing and future research
Breast cancer is a highly heterogeneous disease, at both inter- and intra-tumor levels, and this heterogeneity is a crucial determinant of malignant progression and response to treatments. In addition to genetic diversity and plasticity of cancer cells, the tumor microenvironment contributes to tumor heterogeneity shaping the physical and biological surroundings of the tumor. We recently dissect intra-tumor heterogeneity as a current urgent need to better define breast cancer biology and to develop therapeutic strategies targeting the microenvironment as helpful tools for combined and personalized treatment (Salemme et al., Front Oncol. 2021 PMID: 3377775; Salemme et al., Front Oncol. 2023 PMID: 37265795). We presented evidence that, by its ability to interact with and inhibit β-Catenin in the breast cancer stem cell compartment, p140Cap can orchestrate an anti-tumor immune response, influencing the composition of the TME immune infiltrate to prevent the establishment of a tumor conducive immune environment (Salemme et al., Nat Commun 2023 PMID: 37169737). It is our goal to mechanistically address the relevance of the WNT/β-Catenin pathway in BC, specifically on the TIC compartment, to depict the molecular interplay between p140Cap and BC heterogeneity, and the impact of p140Cap on drug responsiveness. We will offer a comprehensive picture of new opportunities for therapeutic interventions aimed at providing an effective response to counteract tumor progression.
p140Cap family proteins and synaptic plasticity
We have shown that the p140Cap adaptor is highly expressed in neurons, with an enrichment in synaptic structures, where it participates in dendritic spine remodeling (Jaworski et al., Neuron 2009 PMID:19146815; Nat Comm 2013 PMID:23868368; Repetto et al., J Neurosc 2014 PMID:24453341), in synaptogenesis and development of hippocampal inhibitory circuits (Alfieri et al., Front Mol Neurosc 2017 PMID 28713243; Russo et al., Cer Cortex 2017 PMID:29161354) and in drug addiction response (Damez-Werno et al., PNAS 2016 PMID 27506785; Calipari et al., J Neurosc 2019 PMID: 31092585). Our findings provide the molecular and functional dissection of p140Cap as a new active member of a highly dynamic synaptic network involved in memory consolidation, LTP and LTD that are known to be altered in neurological and psychiatric disorders. p140Cap directly binds the GluN2A subunit of NMDAR and modulates the GluN2A-associated molecular network. Moreover, we show for the first time that p140Cap also associates to synaptic lipid rafts and controls the selective recruitment of GluN2A and PSD95 to this specific compartment (Angelini, Morellato et al, JNS, 2022, PMID: 35953295).
Ongoing and future research
Human induced pluripotent stem cells (iPSCs) have been generated by reprogramming adult cells to an embryonic-like state. Since iPSCs can differentiate into neurons, by manipulating the culture conditions, this provides researchers with valuable insights into human neurobiology through an in vitro model. We have already generated iPSC-derived neurons (iN) in p140Cap-/- iN versus WT controls, where we will study the role of p140Cap in human synaptic signaling mainly focusing on molecular pathways related to the actin cytoskeleton and the glutamatergic synaptic activity. Several single-point mutations of p140Cap have been identified in the GeneMatcher platform. These variations give rise to missense variants of p140Cap in subjects presenting neurological disorders such as epilepsy, autism spectrum disorders, and intellectual disorders. Thus, we will reintroduce in the KO iPSCs the WT version of p140Cap and the mutated forms to analyze how the mutants affect the molecular signaling and electrophysiological activity of iN.
Piedmont Region F.E.S.R. 2014-2020 DEFLeCT: Digital tEchnology For Lung Cancer Treatment, co-PI, 70,000 €.
MUR PRIN 2015 2016-2018, “Biological modifiers of HER2 in cancer: from genomic landscape and mechanistic signaling to therapeutic efficacy of HER2 inhibitors”. PI and Project co-ordinator, 369,000 €.
AIRC ID20107 2018-2022“Functional events and molecular mechanisms controlled by p140Cap in curbing the aggressiveness of ERBB2 breast cancers” PI, 664,000 €.
AIRC ID27353 2023-2027, “Functional role of the adaptor protein p140cap in breast cancer: molecular mechanisms and tumor sensitivity to therapy”, PI, 825,000 €.
Health Ministry Finalized Research 2021 2023-2026, “Capturing tumor heterogeneity exploiting a living biobank of ER-positive Breast Cancer Patient-Derived Organoids (PDO)and Xenografts (PDX), to personalize cancer treatment and overcome treatment resistance.”. Co-applicant, 67.500 €.
MUR PRIN 2022 2023-2025, “Protein-protein interactions as regulators of molecular complexes involved in synaptic plasticity: the paradigm of the adaptor protein p140Cap” PI and Project co-ordinator 189.409 €.
MUR PRIN PNRR 2022 2023-2025, Mevalonate pathway and cholesterol modulation by p140Cap and UBIAD1 in breast cancer” PI 232.000 €.
PNRR M4C2-Investimento 1.4-CN00000041 "Finanziato dall'Unione Europea NextGenerationEU" 60.000 €
Publications: https://scholar.google.com/citations?hl=it&user=uv9dveoAAAAJ&view_op=list_works&sortby=pubdate (Link Scopus)
1: Salemme V, Vedelago M, Sarcinella A, Moietta F, Piccolantonio A, Moiso E, Centonze G, Manco M, Guala A, Lamolinara A, Angelini C, Morellato A, Natalini D, Calogero R, Incarnato D, Oliviero S, Conti L, Iezzi M, Tosoni D, Bertalot G, Freddi S, Tucci FA, De Sanctis F, Frusteri C, Ugel S, Bronte V, Cavallo F, Provero P, Gai M, Taverna D, Turco E, Pece S, Defilippi P. p140Cap inhibits β-Catenin in the breast cancer stem cell compartment instructing a protective anti-tumor immune response. Nat Commun. 2023 May 11;14(1):2350. doi: 10.1038/s41467-023-37824-y. PMID: 37169737; PMCID: PMC10175288.
2: Costamagna A, Natalini D, Camacho Leal MDP, Simoni M, Gozzelino L, Cappello P, Novelli F, Ambrogio C, Defilippi P, Turco E, Giovannetti E, Hirsch E, Cabodi S, Martini M. Docking Protein p130Cas Regulates Acinar to Ductal Metaplasia During Pancreatic Adenocarcinoma Development and Pancreatitis. Gastroenterology. 2022 Apr;162(4):1242-1255.e11. doi: 10.1053/j.gastro.2021.12.242. Epub 2021 Dec 17. PMID: 34922945.
3: Angelini C, Morellato A, Alfieri A, Pavinato L, Cravero T, Bianciotto OT, Salemme V, Natalini D, Centonze G, Raspanti A, Garofalo T, Valdembri D, Serini G, Marcantoni A, Becchetti A, Giustetto M, Turco E, Defilippi P. p140Cap regulates the composition and localization of the NMDAR complex in synaptic lipid rafts. J Neurosci. 2022 Aug 10;42(38):7183–200. doi:10.1523/JNEUROSCI.1775-21.2022. Epub ahead of print. PMID: 35953295; PMCID:PMC9512579.
4: Grasso S, Cangelosi D, Chapelle J, Alzona M, Centonze G, Lamolinara A, Salemme V, Angelini C, Morellato A, Saglietto A, Bianchi FT, Cabodi S, Salaroglio IC, Fusella F, Ognibene M, Iezzi M, Pezzolo A, Poli V, Di Cunto F, Eva A, Riganti C, Varesio L, Turco E, Defilippi P. The SRCIN1/p140Cap adaptor protein negatively regulates the aggressiveness of neuroblastoma. Cell Death Differ. 2020 Feb;27(2):790-807. doi: 10.1038/s41418-019-0386-6. Epub 2019 Jul 8.
5: Grasso S, Chapelle J, Salemme V, Aramu S, Russo I, Vitale N, Verdun di Cantogno L, Dallaglio K, Castellano I, Amici A, Centonze G, Sharma N, Lunardi S, Cabodi S, Cavallo F, Lamolinara A, Stramucci L, Moiso E, Provero P, Albini A, Sapino A, Staaf J, Di Fiore PP, Bertalot G, Pece S, Tosoni D, Confalonieri S, Iezzi M, Di Stefano P, Turco E, Defilippi P. The scaffold protein p140Cap limits ERBB2-mediated breast cancer progression interfering with Rac GTPase-controlled circuitries. Nat Commun. 2017 Mar 16;8:14797. doi: 10.1038/ncomms14797.
6: Repetto D, Camera P, Melani R, Morello N, Russo I, Calcagno E, Tomasoni R, Bianchi F, Berto G, Giustetto M, Berardi N, Pizzorusso T, Matteoli M, Di Stefano P, Missler M, Turco E, Di Cunto F, Defilippi P. p140Cap regulates memory and synaptic plasticity through Src-mediated and citron-N-mediated actin reorganization. J Neurosci. 2014 Jan 22;34(4):1542-53. doi:10.1523/JNEUROSCI.2341-13.2014. PMID: 24453341; PMCID: PMC6705312.
7: Morello V, Cabodi S, Sigismund S, Camacho-Leal MP, Repetto D, Volante M,cPapotti M, Turco E, Defilippi P. β1 integrin controls EGFR signaling and tumorigenic properties of lung cancer cells. Oncogene. 2011 Sep 29;30(39):4087-96. doi: 10.1038/onc.2011.107. Epub 2011 Apr 11. PMID: 21478906.
8: Cabodi S, del Pilar Camacho-Leal M, Di Stefano P, Defilippi P. Integrin signalling adaptors: not only figurants in the cancer story. Nat Rev Cancer. 2010 Dec;10(12):858-70. doi: 10.1038/nrc2967. Epub 2010 Nov 24. PMID: 21102636.
9: 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. doi: 10.1038/sj.emboj.7601724. Epub 2007 May 24. PMID: 17525734; PMCID: PMC1894765.
10: 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. doi: 10.1093/emboj/17.22.6622. PMID: 9822606; PMCID: PMC1171008.
