ITT Istituto Toscano Tumori (logo)
Ricerca avanzata
 

Experimental Oncology

Institute of Clinical Physilogy, CNR Siena Unit
c/o Toscana Life Sciences Foundation
via Fiorentina 1 - 53100 Siena
Tel. 0577-1916295
Fax 0577-1916298
website

Principal Investigator: Dr. Caterina Cinti

staff picture   Staff

  • Caterina Cinti, Ph.D, Director of IFC-CNR, Siena Unit, Molecular Biologist (CNR personnel)
  • Roberto Giordano, Molecular Biologist (CNR personnel)
  • Mariela Estenoz, B.S., Cell Biologist (CNR personnel)
  • Rosalba Casula, Administrative (CNR personnel)
  • Anna Rosa Magnano, Ph.D, Electron Microscopist (University of Siena personnel)
  • Dario La Sala, Ph.D, Molecular Biologist ( grant funded personnel)
  • Margherita Cerrone, PhD, Cellular Biologist (grant funded personnel)

The Institute of Clinical Physiology- CNR, Siena unit originated as a branch of the Institute of Clinical Physiology – CNR, Pisa unit, due to an agreement in 2004 between the University of Siena and CNR. It consists of 4 units of CNR personnel employed full-time, two grant funded people and a graduated technician of the University of Siena, which has been located within CNR for years. The Institute of Clinical Physiology- CNR, Siena unit has a central role in basic and technological research in the area of experimental oncology. The principal sources of funding are provided by CNR. Other sources of funding are: MIUR research funds and international funds (NIH and the EC). Scientific production of the IFC-CNR section of Siena culminates in approximately 5 articles a year published in high impact international journals. The IFC-CNR Institute of Siena will be permanently located within the Toscana Life Sciences (TLS) laboratories in Siena Torre Fiorentina’s NOVARTIS scientific complex since July 2007 and will be provided with research laboratories for studies in molecular biology, genetics and electron microscopy.

The main focus of the IFC-CNR section of Siena is in experimental oncology and is concentrated in understanding the pathogenetic mechanism of specific tumor histotypes (such as retinoblastoma, osteosarcoma, lung and breast tumors, chemotherapy drug sensitivity and resistance) with the aim of identifying tumor specific molecular markers and new therapeutic targets. The research concerning this operative unit can be summarized as follows:

Main Research Themes

  1. Development of new therapeutic strategies for multi-drug resistant tumors through gene targeting.
    Main achievements: Drug resistant osteosarcoma cell line has been isolated in our laboratory, starting from a clone of osteosarcoma drug sensitive cells, in order to develop an experimental model for the study of the molecular mechanisms of drug resistant bone tumors. Molecular markers and the regulation mechanisms of genes linked to drug resistant phenotype were identified. Several in vitro experiments of therapeutic strategies were conducted (hypomethylation of DNA and chemotherapy) in order to evaluate possible new therapeutic protocols to cure drug-resistant osteosarcoma (figure 1).
    Current work: We are planning to select new drug-resistant tumor cell lines to use as experimental models in identifying the tumor-specific target protein.
    Future plans: Develop new therapeutic strategies, to be tested both in vitro and in vivo on drug sensitive and resistant tumors, based on the use of demethylating and deacetylating agents alone or in combination with: a) chemotherapy drugs, b) new target specific drugs (synthetic peptides, gene and antibody therapy), c) short-interfering RNAs.
  2. Development of cDNA tumor-specific microarrays for diagnostic use.
    Main achievements: To evaluate the global expression of the genes that characterize each tumor histotype, cDNA microarrays (MWG Human Cancer Array), containing 1853 human genes associated with tumor development and progression, were used to analyze various tumor histotypes (renal carcinoma, medullary thyroid carcinoma, malignant mesothelioma, skin melanoma, three different histotypes of non-Hodgkin lymphoma, retinoblastoma, and osteosarcoma).
    Current work: We are currently analyzing the data obtained from hierarchical clustering analysis which take into consideration the quantity of data regarding differential expression of cDNA microarrays reported by the various cell lines.
    Future plans It will be constructed a gene bank that characterizes each tumor histotype in order to design diagnostic cDNA microarrays. It will be developed a Software which will be able to correlate gene profile with its functional information present in the GenBank.
  3. Animal models for the study of the neoplastic transformation processes and for development of new therapeutic strategies.
    Main achievements: In vitro experiments on retinoblastoma and non-small-cell lung adenocarcinoma cell lines have evidenced some possible mechanisms of gene deregulation. Analyses of promoter methylation (MSP) and gene expression (cDNA microarrays) were conducted on both retinoblastoma and lung adenocarcinoma cell lines. Epigenetic modifications, due to cytosine hypermethylation on the promoters of some genes, are necessary to determine gene silencing and can compete together with other genetic alterations to neoplastic transformation. Treatments with appropriate doses of DNA methyltrasferase (5-AZA-2-deoxycytidine) and/or histon-deacetylase (Tricostatin A) inhibitors reactivate the expression of tumor-suppressor genes suggesting a possible use of these drugs in the therapy of retinoblastoma and lung cancers (figure 2).
    Current work: To identify the epigenetically modified gene pathways involved in neoplastic transformation of retinoblastoma and lung adenocarcinoma, we are analyzing data (hierarchical clustering) obtained from cDNA microarray analysis performed on these two tumor histotypes treated with demethylating /deacetylating agents.
    Future Plans: In order to evaluate in vivo the effect of treatments with demethylating/deacetylating agents alone or in combination with other drugs, and to develop a new therapeutic protocol for retinoblastoma and lung adenocarcinoma, it will be conducted experiments on three different animal models that develop retinoblastoma and one animal model that develops lung adenocarcinoma.
    Animal models of retinoblastoma: 1) female 6-7 week old CD1 nu/nu mice (nude mice) will receive an injection of approximately 106 Y79 or Weri-Rb1 human tumor cells; 2) transgenic mice over-expressing the SV40 large T antigen (SV40-Tag) which develop retina tumors similar to the human form of retinoblastoma; 3)mutant Rb1 murine model that conditionally express inactive Rb1 after intranasal administration of adenoviral vector Ad-Cre and develop the retinoblastoma after some week of infection.
    Animal models of lung adenocarcinoma: mutant LSL-K-Ras G12D murine model that conditionally expresses active K-Ras after intranasal administration of an adenoviral vector (Ad-Cre) and develop an atypical hyperplasia adenomatosa after 2 weeks of infection, a papillary adenoma after 6 weeks, diffuse adenoma at 12 weeks and adenocarcinoma at 16 weeks.

Research Grants

PRINCIPAL INVESTIGATOR (PI)

  • 1999-2000 CNR Project "Characterization of the functional regions of the oncosuppressor gene Rb2/p130 and identification of mutations that cause its inactivation" (CNR annual program)
  • 2000-2001 CNR Project "Characterization of the functional regions of the oncosuppressor gene Rb2/p130 and identification of mutations that cause its inactivation" (CNR annual program)
  • 2001-2002 CNR Project "Characterization of the functional regions of the oncosuppressor gene Rb2/p130 and identification of mutations that cause its inactivation" (CNR annual program)
  • 2001-2003 CNR Project "Role played by the genes in the retinoblastoma (RBs) family and the genes correlated in neoplastic transformation and in embryogenesis" (CNR triennial program)
  • 2002-2003 CRN Project "Role played by the genes in the retinoblastoma (RBs) and the genes correlated in neoplastic transformation and drug resistance" (CNR annual program)
  • 2003-2004 ISPESL-Research Project "Immunological markers of alterations caused by non-ionized radiation"
  • 2003-2008 NIH Project "pRb2/p130: from the mechanisms to gene therapy". (NIH 2 RO1 CA 060999-09)
  • 2006-2008 CNR Drug Project: Innovative therapeutic strategy in the treatment of human neuroblastoma: combination therapy with demethylate drugs and exposure to very low frequency magnetic fields and to the cyclotron resonance frequency.
  • 2006-2008 CNR Drug Project: Clinical implications of epigenetic events carried by the oncosuppressors and DNS repair genes in the genesis of SNC tumors: development of innovative therapeutic systems on murine animal models with retinoblastoma.

CO-PRINCIPAL INVESTIGATOR

  • 1992-1996 CNR Finalized Project Genetic Engineering, sub-project on Mapping and sequencing of the human genome, "Sorting of the X chromosome and its parts using flow cytometry of cellular lines and somatic hybrids (localization of the oligonucleotide of the STS X chromosome library using the PRINS technique)"
  • 1993-1994 IOR Continuing Research Project, "Codivilla Putti" Research Institute, Rizzoli Orthopedic Institute, "In vitro methods to study cartilaginous physiopathology: cellular response and osmotic stress"
  • 1993-1994 IOR Continuing Research Project, "Codivilla Putti" Research Institute, Rizzoli Orthopedic Institute,, "Immunohistochemical study of the relationship between dystrophin, cytoskeleton and cellular membrane"
  • 1993-1994 IOR Continuing Research Project, "Codivilla Putti" Research Institute, Rizzoli Orthopedic Institute, "Study of gene p53 mutations in tumoral osteosarcoma cellular lines"
  • 1993-1994 IOR Continuing Research Project, "Codivilla Putti" Research Institute, Rizzoli Orthopedic Institute, "Localization of the dystrophin gene on metaphasical chromosomes in confocal microscopy and electron microscopy using the PRINS technique"
  • 1994-1995 IOR Continuing Research Project, "Codivilla Putti" Research Institute, Rizzoli Orthopedic Institute,," Use of synthetic oligonucleotides for the identification of single dystrophin gene exons"
  • 1994-1995 IOR Continuing Research Project, "Codivilla Putti" Research Institute, Rizzoli Orthopedic Institute, "Cytochemical investigation on glycosaminoglycan of the cartilagin matrix in dyschondroplasia of mechanical-osmotic stress"
  • 1996-2002 NIH Project "Molecular characterization of one of the members of the retinoblastoma family" (NIH PO1 CA56309)
  • 2000-2006 MURST Project (LAG-CO3), Plans to develop the scientific network, "Study of biomedical and agroalimentary genes of interest"
  • 2001-2003 NIH Project "Functional interaction between HIV-1 products and cell cycle proteins" (NIH 1 R21 CA94802-01)
  • 2002-2003 CNR-MIUR Oncology Strategic Project, Research Program "Preclinical molecular therapy in oncology"
  • 2003-2004 Strategic Project of in the Biomedical Area, Ministry of Health, "New combination therapeutic strategies: DNA hypomethylates and bioimmunotherapy".
  • 2004-2006 AIRC Project 2004 "ER-transcriptional regulation by pRB proteins: potential prognostic markers and therapeutic targets in breast cancer"
  • 2004-2006 Monte dei Paschi di Siena (MPS) Project 2004: "Role of pRb2/p130 in the genesis of medulloblastoma".

Publications

  1. P.P.Claudio, C. Cinti, A. Giordano. Application of the primer in situ DNA synthesis (PRINS) technique to titer recombinant virus and evaluation of the efficiency of viral transduction. Anal. Biochem. 29: 96-101 (2001).
  2. Zamparelli A, Masciullo V, Bovicelli A, Santini D, Ferrandina G, Minimo C, Terzano P, Costa S, Cinti C, Ceccarelli C, Mancuso S, Scambia G, Bovicelli L, Giordano A. Expression of cell-cycle-associated proteins pRb2/p130 and p27kip in vulvar squamous cell carcinomas. Hum Pathol. 32:4-9. (2001).
  3. N. Zini, C. Trimarchi, P.P. Claudio, P. Stiegler, Marinelli F, M.C. Maltarello, D. La Sala, G. De Falco, G. Russo, G. Ammirati, A. Giordano, N.M. Maraldi, C. Cinti. pRb2/p130 and p107 control cell growth by multiple strategies and in association with different compartments within the nucleus. J. Cell. Physiol. 189: 34-44 (2001).
  4. Maraldi NM, Giordano A, Manzoli L, Falconi M, Pol AD, Cinti C. Genetic alterations at the nuclear localization signal of the RB2/p130 gene occur in lymphoid tumor but not in osteosarcoma cell lines. Adv Enzyme Regul. 41:31-55 (2001).
  5. Commentary by C. Cinti, M. Macaluso. Risk of endometrial carcinoma associated with BRCA mutation. J. Women's Oncol. Review. 1: 145 (2001).
  6. Commentary by C. Cinti and C. Kumar. E-cadherin and alfa-catenin expression during tumor progression of cervical carcinoma. J. Women's Oncol. Review. 1(2): 137 (2001).
  7. Commentary by C. Trimarchi and C. Cinti. Role of vascular endothelial growth factor in the stimulation of cellular invasion and signaling of breast cancer cell. J. Women's Oncol. Review. 1(2): 139. (2001).
  8. Claudio PP, Zamparelli A, Zupi A, Califano L, Bellan C, Minimo C, Howard C, Micheli P, Cinti C, Leoncini L, De Rosa G, Giardino C, Giordano A. Expression of cell-cycle regulated prb2/p130, p107, E2F4, p27 and of PCNA in salivary gland tumors. Prognostic and diagnostic implications. Rivista Italiana di Chirurgia Maxillo-Facciale XII . 1:37-48 (2001)
  9. Pittoggi, C., Magnano, A.R., Sciamanna, I., Giordano, R., Lorenzini, R. and Spadafora, C. (2001) - Specific localization of transcription factors in the chromatin of mouse mature spermatozoa. Mol Reprod Dev. 60(1): 97-106. Wiley-Liss, New York, USA.
  10. C. Cinti, L. Stuppia, N.M. Maraldi. Combined use of PRINS and fish in the study of dystrophin gene. Am. J. Medic. Genet. 107: 115-118 (2002).
  11. C. Bellan, S. Lazzi, M. Zazzi, AV. Lalinga, N. Palummo, P. Galieni, T. Marafioti, T. Tonini, C.Cinti, L. Leoncini, S.Pileri, P.Tosi. Immunoglobulin gene rearrangement analysis in composite Hodgkin's disease and large B-cell lymphoma: evidence for receptor revision of immunoglobulin heavy chain variable region genes in Hodgkin-Reed-Sternberg cells? Diagnostic Mol. Pathol. 11: 2-8 (2002).
  12. Macaluso M, Russo G, Cinti C, Russo A. The Ras family genes: an interesting link between cell cycle and cancer. J.Cell.Physiol. 192: 125-130 (2002). Review
  13. S. Lazzi, C Bellan, G De Falco, C Cinti, F Ferrari, A Nyongo, PP Claudio, GM Tosi, R Vatti, A Gloghini, A Carbone, A Giordano, L Leoncini and P Tosi.Expression of RB2/p130 tumor suppressor gene in AIDS-related non-Hodgkin's lymphomas. Implications for disease pathogenesis. Human Pathol 33:723-731(2002)
  14. C Bellan, G De Falco, GM Tosi, S Lazzi, F Ferrari, G Morbini, S Bartolomei, P Toti, P Mangiavacchi, G Cevenini, C Trimarchi, C Cinti, A Giordano, L Leoncini, P Tosi H Cottier. Missing expression of pRb2/p130 in human retinoblastomas is associated with reduced apoptosis and lesser differentiation. Inv.Ophthal.Visual Sci. 43:3602-3608 (2002).
  15. B. Baccetti, S.Capitani, G. Collodel, M. Estenoz, L. Gambera and P. Piomboni. Infertile spermatozoa in a human carrier of robertsonian translocation 14;22. Fertil. Steril., 78(5): 1127-1130 (2002).
  16. La Sala D., Macaluso M., Trimarchi C., Giordano A., Cinti C. Triggering of p73-dependent apoptosis in osteosarcoma is under the control of E2Fs-pRb2/p130 complexes. Oncogene 22: 3518-3529 (2003).
  17. M Macaluso, C Cinti, G Russo, A Russo & A.Giordano. pRb2/p130-E2F4/5-HDAC1-SUV39H1-DNMT1 multimolecular complexes mediate the transcription of Estrogen Receptor-α in breast cancer. Oncogene 22:3511-3517 (2003).
  18. G. De Falco, C. Bellan, S. Lazzi, P.P. Claudio, D. La Sala, C. Cinti, P. Tosi, A. Giordano and L. Leoncini. The interaction between HIV-1 Tat and pRb2/p130: a possible mechanism in the pathogenesis of AIDS-related neoplasms. Oncogene 22: 6214-6219 (2003).
  19. D.-D Gan, M. Macaluso, C. Cinti, A. Giordano. How a normal human cell becomes a cancer cell. J.Exp.Clin.Cancer Res. 22: 421-427 (2003).
  20. Sciamanna, I., Barberi, L., Martire, A., Pittoggi, C., Beraldi R., Giordano, R., Magnano, A.R., Hodgson, C. and Spadafora, C. (2003) – Sperm endogenous reverse transcriptase as mediator of new genetic information. Biochem. Bioph. Res. Co. 312: 1039-1046. Elsevier Science Ireland Ltd
  21. Marinelli F., La Sala D., Cicciotti G., Cattini L., Trimarchi C., Putti S., Zamparelli A., Giuliani L., Tomassetti G. and Cinti C. Exposure to 900 MHz electromagnetic field induces an unbalance between pro-apoptotic and pro-survival signals in T-lymphoblastoid leukemia CCRF-CEM cells. J.Cell.Physiol. 198:324-332 (2004)
  22. Trimarchi C, La Sala D, Zamparelli A, Cinti C. Detection of apoptotic deoxyribonucleic acid break by in situ nick translation. Methods Mol Biol. 285:113-8. (2004)
  23. Riccio M, Dembic M, Cinti C, Santi S. Multifluorescence labeling and colocalization analyses. Methods Mol Biol. 285:171-7. (2004)
  24. Zini N, Solimando L, Cinti C, Maraldi NM. Single and double colloidal gold labeling in postembedding immunoelectron microscopy. Methods Mol Biol. 285:161-9. (2004)
  25. White MK, Cinti C. A morphologic approach to detect apoptosis based on electron microscopy. Methods Mol Biol. 285:105-11. (2004)
  26. Macaluso M, Montanari M, Cinti C and Giordano A. Modulation of cell cycle components by epigenetic and genetic events. Sem.Oncol. 32: 452-457 (2005).
  27. G.M. Tosi , C. Trimarchi , M. Macaluso , D. La Sala, A. Ciccodicola , S. Lazzi, M. Massaro-Giordano, A. Caporossi, A. Giordano and C. Cinti. Genetic and epigenetic alterations of RB2/p130 tumor suppressor gene in human sporadic retinoblastoma: implications for pathogenesis and therapeutic approach. Oncogene 24: 5827-5836. (2005).
  28. C. Cinti, M.Macaluso and A.Giordano. Tumor-specific exon 1 mutations could be the "hit event" predisposing Rb2/p130 gene to epigenetic silencing in lung cancer. Oncogene 24: 5821-5826 (2005).
  29. B. Baccetti, G. Collodel, M. Estenoz, D. Manca, E. Moretti, P. Piomboni. Gene deletions in an infertile man with sperm fibrous sheath dysplasia. Hum. Reprod. 20(10):2790-4 (2005).
  30. G. Collodel, E. Moretti, S. Capitani, P. Piomboni, C. Anichini, M. Estenoz, B. Baccetti. TEM, FISH and molecular studies in infertile men with pericentric inversion of chromosome 9. 38(4):122-7 (2006).
  31. Pittoggi C., Beraldi R., Sciamanna I., Barberi L., Giordano R, Magnano A.R., Torosantucci L., Pescarmona E. and Spadafora C - Generation of biologically active retro-genes upon interaction of mouse spermatozoa with exogenous DNA. Mol Reprod Dev. 2006 Oct;73(10):1239-46 (2006)
  32. Roberti A, La Sala D. and Cinti C. Multiple genetic and epigenetic interacting mechanisms contribute to clonally selection of drug resistant tumors: current views and new therapeutic prospective. J.Cell.Physiol. 207: 571-581 (2006).

Books chapters:

  • R. Santoro, F.Marinelli, G. Turchetti, N.Zini, K. Scotlandi, E. Falcieri, C. Cinti, N.M. Maraldi. From Fractals in biology and medicine, Fractal analysis of chromatin during apoptosis. Edit by GA Losa, D Merlini, TF Nonnenmacher, ER Weibel, BertelsmannSpring Publishing Group, Vol 3, p 77-84 (2002).
  • Leoncini L, C Bellan, Cinti C, Giordano A. From Cancer Drug Discovery and development: Cell cycle inhibitors in cancer therapy, Tumor suppressor genes as diagnostic tools. Edit by A.Giordano and KJ Soprano, Humana Press Inc.,Totowa, NJ. Chapter 4, p: 49-82 (2002).
  • Cinti C, Trimarchi C, Giordano A. In G1 phase progression, G1 progression and apoptosis. Edited by J. Boonstra, Landes Bioscience publishers, Eurekah.com and Kluwer Academic/Plenum Publishers. Chapter 12, p: 199-235 (2003).
  • M.K. White, T.G. Schneider, C Cinti. In Cyclin and cyclin dependent kinase protocols, A morphologic approach to detect apoptosis based on electron microscopy. Edit by A.Giordano and G. Romano, Humana Press Inc.,Totowa, NJ. Chapter 13, p: 105-111 (2004)
  • C.Trimarchi, D. La Sala, A. Zamparelli, C. Cinti. In Cyclin and cyclin dependent kinase protocols, Detection of apoptotic DNA break by in situ Nick Translation. Edit by A.Giordano and G. Romano, Humana Press Inc.,Totowa, NJ. Chapter 14, p: 113-118 (2004)
  • N. Zini, L. Solimando, C. Cinti, N.M. Maraldi. In Cyclin and cyclin dependent kinase protocols, Single and double colloidal gold labeling in post-embedding immunoelectron microscopy. Edit by A.Giordano and G. Romano, Humana Press Inc.,Totowa, NJ. Chapter 21, p: 161-169 (2004)
  • M. Riccio, M. Dembic, C. Cinti, S. Santi. In Cyclin and cyclin dependent kinase protocol, Multifluorescence labeling and co-localization analyses. Edit by A.Giordano and G. Romano, Humana Press Inc.,Totowa, NJ. Chapter 22, p: 171-177 (2004)
  • L. Stuppia, D. La Sala and C. Cinti. In PRINS and In Situ PCR Protocols Combined FISH and PRINS labeling for the analysis of the dystrophin gene. Edit by F. Pellestor. Humana Press Inc.,Totowa, NJ. Charter 11, p: 115-122 (2006).

Collaborations

  • Department of Oncology, University Hospital of Siena:
    1) Division of Human Pathology;
    2) Division of Medical Oncology and Immunotherapy
  • Institute of Biomedical Technology (ITB), CNR Bari;
  • Institute of Neuroscience (IN), CNR Pisa;
  • Institute of Neurobiology and Molecular Medicine(INMM), CNR Roma;
  • Istituto Superiore di Sanità, Roma;
  • Institute of Regina Elena, Roma;
  • Joan C. Edwards School of Medicine, Marshall University, Huntington, USA;

^ torna all'inizio