Target Identification and Validation Unit


This part of the Program is based on the assumption that there is a fundamental biological heterogeneity among tumors, and these differences must be understood and exploited to identify cellular pathways governing the biology of the tumor within a specific patient, that can be targeted pharmacologically. In our view, rather than taking a "descriptive" approach to the anatomy of tumors, a "functional" strategy has better chances to lead to the identification of immediately validated targets. To this end we have generated an in vivo RNAi screening platform with the aim to screen a large cohort of patients' samples transplanted in immunocompromised animals (NSG mice) with lentiviral-based shRNA libraries of epigenetic and metabolic targets and kinases.

  • Research projects

    Our interest is focused on aggressive cancers for which patients have few effective treatment options, namely metastatic melanoma, metastatic breast carcinoma resistant to conventional therapy and adenocarcinoma of the pancreas. Freshly ex-vivo explanted human tumors are transplantated in NSG mice mainly orthotopically, according to well-established techniques.
    Patient-derived xenografts (PDX) phenocopy the heterogeneity and the complexity of the tumor of origin. Moreover, an extensive immunophenotypic characterization of serial transplantation of the human tumors has shown a stably reproducible propagation of the original tumor. A panel of human tumor markers, as well as whole exome sequencing and epigenetic and proteomic profile of the tumors, are used to better characterize the human phenocopy in the mouse. Our aim is to stratify our cohort of PDXs according to: i) in vivo growth properties; ii) phenotypic markers; iii) genetic lesions; iv) characterization of intracellular signaling networks and v) epigenetic profile. At the end we might be able to generate a prognostic signature of the patient and the most suitable model for RNAi in vivo screenings.
    To set up the in vivo RNAi screening in our model systems we are investigating the frequency of tumor initiating cells in the tumor samples and the interaction of the tumor cells with the microenvironment. We have set up the proper amplification and sequencing technique to evaluate hairpins’ representation in the tumor and analyse and validate target genes and pathways.

    Melanoma is an aggressive disease with high metastatic potential and resistance to cytotoxic agents. The molecular mechanisms involved in the progression of the malignancy and the genetic markers associated with metastatic melanoma dissemination and the acquisition of chemoresistance are only beginning to be defined. An understanding of the underlying molecular biology of melanoma provides a necessary basis to enable the generation of more effective therapeutic modalities. Our interest is to understand which are the molecular pathways involved in melanoma-genesis by an in vivo approach. In particular, we will pursue in vivo RNAi screens in metastatic melanoma PDXs to identify druggable genes that are essential for melanoma growth and progression.
    Breast cancer is not a single disease. Deregulation of specific gene pathways is associated with different sensitivity to chemotherapy in various subtypes. Some clinically relevant molecular aberrations are identified in a subset of metastatic breast cancer patient population or also, due to tumor heterogeneity, within subclones of tumor metastases. Triple negative (TN), luminal B (LB) and HER2 positive (H+) breast cancer are characterized by high risk of relapse following adjuvant therapy, resulting in a rapidly fatal clinical course. To identify genetic alterations and pathways involved in tumor resistance we will couple in vivo RNAi-based screening in a xeno-transplantation setting obtained from tumor metastases to exome sequencing on tumor samples from metastases, archival primary and xenograft.

  • Publications

    • Aladowicz E, Ferro L, Vitali GC, Venditti E, Fornasari L, Lanfrancone L: Molecular networks in melanoma invasion and metastasis. Future Oncol. 2013 May;9(5):713-26.
    • Turco MY, Furia L, Dietze A, Fernandez Diaz L, Ronzoni S, Sciullo A, Simeone A, Constam D, Faretta M, Lanfrancone L: Cellular heterogeneity during embryonic stem cell differentiation to epiblast stem cells is revealed by the ShcD/RaLP adaptor protein. Stem Cells. 2012 Nov;30(11):2423-36.
    • Bosotti R, Carpinelli P, Healy S, Locatelli G, Cappella P, Lanfrancone L, Calogero R, Moll J, Isacchi A: Transcriptional analysis of the Aurora inhibitor Danusertib leading to biomarker identification in TP53 wild type cells. Gene. 2012 Feb 25;494(2):202-8.
    • Licciulli S, Luise C, Scafetta G, Capra M, Giardina G, Nuciforo P, Bosari S, Viale G, Mazzarol G, Tonelli C, Lanfrancone L, Alcalay M: Pirin inhibits cellular senescence in melanocytic cells. Am J Pathol. 2011 May;178(5):2397-406.
    • Licciulli S, Luise C, Zanardi A, Giorgetti L, Viale G, Lanfrancone L, Carbone R, Alcalay M: Pirin delocalization in melanoma progression identified by high content immuno-detection based approaches. BMC Cell Biol. 2010 Jan 20;11:5.
    • Pasini L, Turco MY, Luzi L, Aladowicz E, Fagiani E, Lanfrancone L: Melanoma: targeting signaling pathways and RaLP. Expert Opin Ther Targets. 2009 Jan;13(1):93-104.
    • Santoriello C, Deflorian G, Pezzimenti F, Kawakami K, Lanfrancone L, d'Adda di Fagagna F, Mione M: Expression of H-RASV12 in a zebrafish model of Costello syndrome causes cellular senescence in adult proliferating cells. Dis Model Mech. 2009 Jan-Feb;2(1-2):56-67.



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