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LABORATORY OF: RNA Technology 2
CONTACT PERSON: Dr.Simona Zupo (IST)
Phone +39 010 5737582 E-mail: simona.zupo@istge.it

Description of Laboratory and Expertise:

Since 1996 Simonetta Zupo and her group have been working on the pathophysiology of B lymphocytes, conducting studies both on the normal B lymphocyte populations and B lymphoproliferative diseases. Part of her studies have regarded the phenotypical and functional characterization of B cell-Chronic lymphocytic leukaemia (B-CLL). Although the B-CLL cells are morphologically homogeneous, the disease has a heterogeneous clinical course. This group contributed to the identification of subsets of CLL patients based on phenotypical, molecular and functional features. Recently, it has been demonstrated that some of these characteristics can be considered new prognostic markers. In the last 5 years Simonetta Zupo has been collaborating with Dr. CM Croce studying the role of miRNA on the pathogenesis of B-CLL. These studies provided evidences that some miRNA (miR15a and miR16-1) are involved in the deregulation of the apoptotic process in the leukaemic B cells.

Abstract of Activities:

miRNA are a small ncRNA class of genes conserved in sequence between distantly related organisms , suggesting that these molecules participate in essential processes. The functions of miRNA are highly varied and include the modulation of hematopoietic lineage differentiation in mammals. Several recent data proved that the alterations in microRNA genes play a critical role in the pathophysiology of many human cancers. Loss or amplification of miRNA genes has been reported in a variety of cancers including B-CLL and altered patterns of miRNA expression may affect cell cycle or survival programs (fig 1). Recently, Croce’s group with our collaboration demonstrated that the levels of miR-15 and miR-16 inversely correlate with the bcl2 protein expression and that bcl2 repression by these miR induces apoptosis in leukemic cells. The deletion/mutation of these microRNA observed in B-CLL cells is responsible for the overexpression of bcl2 in these neoplastic cells and can be considered pathogenetic lesions in CLL (Figs.1, 2).
Starting from this knowledge, this group intends to investigate the following points: 1) the role of miRNA in the pathogenesis of other lymphoproliferative diseases, besides B-CLL. 2) the role of the altered miRNA in the mechanisms of post-transcriptional regulation of BCL2-mRNA in B-CLL cells compared with those observed in their normal counterpart B populations.

Detailed Research Activities:

The studies about miRNA profile in lymphoproliferative diseases are based on the comparison between the patterns obtained in normal B cell counterparts and those obtained in the leukemic cells of the specific diseases. Therefore, the first step is to purify normal B cell subsets considered the normal counterparts of some lymphoma from tonsil and spleen. Briefly, a FACS sorting methodology or magnetic beads methods allows us to separate the normal B cells in three different populations: Follicular mantle (FM), Germinal Center (GC) and Marginal Zone B cells (MZ). In addition, both resting and activated MZ cells and plasmacells will be prepared. At the same time, leukemic cells will be purified from biopsies or other materials of patients affected by lymphoma. The main kind of lymphoma that we intend to investigate will be: Follicular lymphoma, mantle cell lymphoma, myeloma , B-CLL. RNA and protein will be prepared from these cellular materials and miRNA patterns will be detected.

Applications and Developments:

Several chromosomal translocations and other genomic abnormalities have a clear diagnostic and prognostic significance, as well some gene expression profiles. In the same way, the altered miRNA expression in lymphoproliferative diseases could be clinically exploited.
The ultimate aim of these researches is to provide new tools for diagnosis and prognosis of the these diseases based on miRNA profiling. Moreover, the potential use of miRNA in cancer therapy is now being explored. The theoretical rationale is based on the fact that miRNA are natural antisense interactors that regulate many genes involved in survival and proliferation. There are several examples of this kind of translational research, such as chronic myelogenous leukaemia and Gleevec, a drug that inhibits the kinase involved in the disease-specific chromosomic translocation BCR-ABL.

Ongoing collaborations:

Croce CM, M.D., is Director of the Department of Molecular Virology , Immunology and Medical genetics, Colleges of Medicine and Public Health and Cancer Center. Ohio State University, Columbia, Ohio.
His main interest is to investigate the function of miRNAs, in particular the micro RNA signatures in human normal and tumor cells.

Most recent and significant publications

Piccaluga PP, Agostinelli C, Califano A, Carbone A, Fantoni L, Ferrari S, Gazzola A, Gloghini A, Righi S, Rossi M, Tagliafico E, Zinzani PL, Zupo S, Baccarani M, Pileri SA. Gene expression analysis of angioimmunoblastic lymphoma indicates derivation from T follicular helper cells and vascular endothelial growth factor deregulation.
Cancer Res. 2007 Nov 15;67(22):10703-10.

Calin GA, Liu CG, Ferracin M, Hyslop T, Spizzo R, Sevignani C, Fabbri M, Cimmino A, Lee EJ, Wojcik SE, Shimizu M, Tili E, Rossi S, Taccioli C, Pichiorri F, Liu X, Zupo S, Herlea V, Gramantieri L, Lanza G, Alder H, Rassenti L, Volinia S, Schmittgen TD, Kipps TJ, Negrini M, Croce CM.
Ultraconserved regions encoding ncRNAs are altered in human leukemias and carcinomas.
Cancer Cell. 2007 Sep;12(3):215-29

Gargiulo L, Lastraioli S, Cerruti G, Serra M, Loiacono F, Zupo S, Luzzatto L, Notaro R Highly homologous T-cell receptor beta sequences support a common target for autoreactive T cells in most patients with paroxysmal nocturnal hemoglobinuria.
Blood. 2007 Jun 1;109(11):5036-42. Epub 2007 Feb 6.

Ropolo M, Degan P, D'Errico M, Dogliotti E, Zupo S, Poggi A, Frosina G, Complementation of the oxidatively damaged DNA repair defect in Cockayne syndrome A and B cells by Escherichia coli formamidopyrimidine DNA glycosylase.
Free Radic Biol Med. 2007 Jun 15;42(12):1807-17. Epub 2007 Mar 12.

Gene expression analysis of peripheral T cell lymphoma, unspecified, reveals distinct profiles and new potential therapeutic targets.
J Clin Invest. 2007 Mar;117(3):823-34. Epub 2007 Feb 15

Dominietto A.,Pozzi S., Miglino M., Albarracin F., Piaggio G., Bertolotti F., Grasso R., Zupo S., Raiola AM., Gobbi M., Frassoni F., Bacigalupo A. Donor lymphocyte infusions for the treatment of minimal residual disease in acute leukemia.
Blood. 2007 Jun 1;109(11):5063-4.

Ropolo M, Geroldi A., Degan P., Andreotti V., Zupo S., Poggi A., Reed A., Kelley MR., Frosina G. Accelerated repair and reduced mutagenicity of oxidative DNA damage in human bladder cells expressing the E. coli FPG protein.
Int J Cancer. 2006 Apr 1;118(7):1628-34

De Totero D., Meazza T., Zupo S., Cutrona G., Matis S.,Colombo M., Balleari E., Pierri I., Fabbi M., Capaia M., Azzarone B., Gobbi M., Ferrarini M., Ferrini S. Interleukin-21 receptor (IL-21R) is up-regulated by CD40 triggering and mediates proapoptotic signals in chronic lymphocytic leukemia B cells.
Blood. 2006 May 1;107(9):3708-15. Epub 2006 Jan 3.

Piccaluga PP., Agostinelli C., Zupo S:, Facchetti F., Falini B., Ferrarini M., Gallamini A., Novero D., Paulli M., Zinzani PL., Dalla favera R. Pileri S Gene expression analysis of peripheral T-cell lymphoma not otherwise specified reveals two distinct subgroups and recurrente PDGFR-alpha deregulation Heamatologica reports 2006 2:66-67

Cimmino A.-Calin G.-Fabbri M.-Iorio M.-Ferracin M.-Shimizu M.-Wojcik S.-Aqeilan R.-Zupo S.-Dono M.-Rassenti L.- Alder H.-Volinia S.-Liu C.-Kipps T.-Negrini M.-Croce C. Mir/15 And Mir/16 Induce Apoptosis By Targeting Bcl2. Proc. Natl. Acad. Sci. Usa 2005 102:13944/13949

Messmer B.-Messmer D.-Allen S.-Kolitz J.-Kudalkar P.-Cesar D.-Murphy E.-Koduru P.-Ferrarini M.- Zupo S.-Cutrona G.-Damle R.-Wasil T.-Rai K.-Hellerstein M.-Chiorazzi N. In vivo measurements document the dynamic cellular kinetics of chronic lymphocytic leukemia B cells.
J Clin Invest. 2005 Mar;115(3):755-64.

Calin G.-Liu C.-Sevignani C.-Ferracin M.-Felli N.-Dumitru C.-Shimizu M.-Cimmino A.-Zupo S.-Dono M. -Dell'aquila M.-Alder H.-Rassenti L.-Kipps T.-Bullrich F.-Negrini M.-Croce C. MicroRNA profiling reveals distinct signatures in B cell chronic lymphocytic leukemias.
Proc Natl Acad Sci U S A. 2004 Aug 10;101(32):11755-60

Liu C.-Calin G.-Meloon B.-Gamliel N.-Sevignani C.-Ferracin M.-Dumitru C.-Shimizu M.-Zupo S.-Dono M. -Alder H.-Bullrich F.-Negrini M.-Croce C. An oligonucleotide microchip for genome-wide microRNA profiling in human and mouse tissues.
Proc Natl Acad Sci U S A. 2004 Jun 29;101(26):9740-4. Epub 2004 Jun 21