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ABSTRACT: Accumulating evidence supports the potential of proteasome inhibitors as immunosuppressants. Proteasome inhibitors interfere with antigen processing and presentation, as well as with the signaling cascades involved in immune cell function and survival. Both myeloma and healthy plasma cells appear to be highly susceptible to proteasome inhibitors due to impaired proteasomal activity in both cell types. As a consequence, these agents can be used to reduce antibody production and thus prevent antibody-induced tissue damage. Several clinical studies have explored the potential of bortezomib, a peptide boronate proteasome inhibitor, for treating immune disorders, such as antibody-mediated organ rejection and graft-versus-host disease (GVHD), with encouraging results. Here, we discuss the biological rationale for the use of proteasome inhibitors as immunosuppressive agents and review the clinical experience with bortezomib in immune-mediated diseases.
Seminars in Hematology 07/2012; 49(3):270-6. · 3.99 Impact Factor
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Claudia Bello,
Giovanna Dal Bello,
Michele Cea,
Aimable Nahimana,
Dominique Aubry,
Anna Garuti,
Giulia Motta, Eva Moran,
Floriana Fruscione,
Paolo Pronzato, [......],
Franco Patrone,
Alberto Ballestrero,
Marc Dupuis,
Bernard Sordat,
Kaspar Zimmermann,
Jacqueline Loretan,
Markus Wartmann,
Michel A Duchosal,
Alessio Nencioni,
Pierre Vogel
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ABSTRACT: New derivatives of 1,4-dideoxy-1,4-imino-D-ribitol have been prepared and evaluated for their cytotoxicity on solid and haematological malignancies. 1,4-Dideoxy-5-O-[(9Z)-octadec-9-en-1-yl]-1,4-imino-D-ribitol (13, IC(50) ∼2 μM) and its C(18)-analogues (IC(50) <10 μM) are cytotoxic toward SKBR3 (breast cancer) cells. 13 also inhibits (IC(50) ∼8 μM) growth of JURKAT cells.
Bioorganic & medicinal chemistry 07/2011; 19(24):7720-7. · 2.82 Impact Factor
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Michele Cea,
Debora Soncini,
Floriana Fruscione,
Lizzia Raffaghello,
Anna Garuti,
Laura Emionite, Eva Moran,
Mirko Magnone,
Gabriele Zoppoli,
Daniele Reverberi, [......],
Salvatore Casciaro,
Ivana Pierri,
Gianluca Damonte,
Filippo Ansaldi,
Marco Gobbi,
Vito Pistoia,
Alberto Ballestrero,
Franco Patrone,
Santina Bruzzone,
Alessio Nencioni
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ABSTRACT: Aberrant histone deacetylase (HDAC) activity is frequent in human leukemias. However, while classical, NAD(+)-independent HDACs are an established therapeutic target, the relevance of NAD(+)-dependent HDACs (sirtuins) in leukemia treatment remains unclear. Here, we assessed the antileukemic activity of sirtuin inhibitors and of the NAD(+)-lowering drug FK866, alone and in combination with traditional HDAC inhibitors. Primary leukemia cells, leukemia cell lines, healthy leukocytes and hematopoietic progenitors were treated with sirtuin inhibitors (sirtinol, cambinol, EX527) and with FK866, with or without addition of the HDAC inhibitors valproic acid, sodium butyrate, and vorinostat. Cell death was quantified by propidium iodide cell staining and subsequent flow-cytometry. Apoptosis induction was monitored by cell staining with FITC-Annexin-V/propidium iodide or with TMRE followed by flow-cytometric analysis, and by measuring caspase3/7 activity. Intracellular Bax was detected by flow-cytometry and western blotting. Cellular NAD(+) levels were measured by enzymatic cycling assays. Bax was overexpressed by retroviral transduction. Bax and SIRT1 were silenced by RNA-interference. Sirtuin inhibitors and FK866 synergistically enhanced HDAC inhibitor activity in leukemia cells, but not in healthy leukocytes and hematopoietic progenitors. In leukemia cells, HDAC inhibitors were found to induce upregulation of Bax, a pro-apoptotic Bcl2 family-member whose translocation to mitochondria is normally prevented by SIRT1. As a result, leukemia cells become sensitized to sirtuin inhibitor-induced apoptosis. In conclusion, NAD(+)-independent HDACs and sirtuins cooperate in leukemia cells to avoid apoptosis. Combining sirtuin with HDAC inhibitors results in synergistic antileukemic activity that could be therapeutically exploited.
PLoS ONE 01/2011; 6(7):e22739. · 4.09 Impact Factor
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Gabriele Zoppoli,
Michele Cea,
Debora Soncini,
Floriana Fruscione,
Justine Rudner, Eva Moran,
Irene Caffa,
Davide Bedognetti,
Giulia Motta,
Riccardo Ghio,
Fabio Ferrando,
Alberto Ballestrero,
Silvio Parodi,
Claus Belka,
Franco Patrone,
Santina Bruzzone,
Alessio Nencioni
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ABSTRACT: The nicotinamide phosphoribosyltransferase (Nampt) inhibitor APO866 depletes intracellular nicotinamide adenine dinucleotide (NAD(+)) and shows promising anticancer activity in preclinical studies. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) binds to plasma membrane receptors DR4 and DR5 and induces apoptosis via caspase-8 and -10. Here we have explored the interaction between APO866 and TRAIL in leukemia cell lines and in primary B-cell chronic lymphocytic leukemia cells.
Cells were treated with APO866, TRAIL, or their combination. Viability and mitochondrial transmembrane potential (ΔΨ(m)) were determined by cell staining with propidium iodide and tetramethylrhodamine ethyl ester, respectively, and flow cytometry. Nampt and γ-tubulin levels, as well as caspase-3 cleavage were detected by immunoblotting. DR4 and DR5 expression were assessed by immunostaining and flow cytometry. Caspases were inhibited with zVAD-FMK and zDEVD-FMK; autophagy with 3-methyladenine, LY294002, and wortmannin. Intracellular NAD(+) and adenosine triphosphate (ATP) were measured by cycling assays and high-performance liquid chromatography (HPLC), respectively.
APO866 induced NAD(+) depletion, ΔΨ(m) dissipation, and ATP shortage in leukemia cells, thereby leading to autophagic cell death. TRAIL induced caspase-dependent apoptosis. TRAIL addition to APO866 synergistically increased its activity in leukemia cells by enhancing NAD(+) depletion, ΔΨ(m) dissipation, and ATP shortage. No DR5 upregulation at the cell surface in response to APO866 was observed. Remarkably, in healthy leukocytes APO866 and TRAIL were poorly active and failed to show any cooperation.
Activation of the extrinsic apoptotic cascade with TRAIL selectively amplifies the sequelae of Nampt inhibition in leukemia cells, and appears as a promising strategy to enhance APO866 activity in hematological malignancies.
Experimental hematology 11/2010; 38(11):979-88. · 3.11 Impact Factor
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Michele Cea,
Gabriella Cirmena,
Anna Garuti,
Ilaria Rocco,
Claudia Palermo,
Antonia Cagnetta, Eva Moran,
Nicoletta Colombo,
Raffaella Grasso,
Giuseppina Fugazza,
Marco Gobbi,
Alessio Nencioni,
Alberto Ballestrero,
Franco Patrone
Leukemia research 05/2010; 34(9):e240-2. · 2.36 Impact Factor
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Claudia Bello,
Michele Cea,
Giovanna Dal Bello,
Anna Garuti,
Ilaria Rocco,
Gabriella Cirmena, Eva Moran,
Aimable Nahimana,
Michel A Duchosal,
Floriana Fruscione,
Paolo Pronzato,
Francesco Grossi,
Franco Patrone,
Alberto Ballestrero,
Marc Dupuis,
Bernard Sordat,
Alessio Nencioni,
Pierre Vogel
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ABSTRACT: Novel alpha-mannosidase inhibitors of the type (2R,3R,4S)-2-({[(1R)-2-hydroxy-1-arylethyl]amino}methyl)pyrrolidine-3,4-diol have been prepared and assayed for their anticancer activities. Compound 30 with the aryl group=4-trifluoromethylbiphenyl inhibits the proliferation of primary cells and cell lines of different origins, irrespective of Bcl-2 expression levels, inducing a G2/Mcell cycle arrest and by modification of genes involved in cell cycle progression and survival.
Bioorganic & medicinal chemistry 03/2010; 18(9):3320-34. · 2.82 Impact Factor
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Gabriele Zoppoli, Eva Moran,
Debora Soncini,
Michele Cea,
Anna Garuti,
Ilaria Rocco,
Gabriella Cirmena,
Valentina Grillo,
Luca Bagnasco,
Giancarlo Icardi,
Filippo Ansaldi,
Silvio Parodi,
Franco Patrone,
Alberto Ballestrero,
Alessio Nencioni
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ABSTRACT: Lapatinib, a dual HER2 and EGFR tyrosine kinase inhibitor is highly active in HER2+ breast cancer. However, its efficacy is limited by either primary or acquired resistance. Although mutations in ras genes are rarely found in breast cancer, H-ras overexpression is frequently observed. Moreover, genetic alterations that do not directly involve ras such as Brk amplification, ultimately result in increased ras signaling. Using SKBR3 cells, a HER2+ breast cancer cell line that is naturally devoid of mutations in PI3KCA, PTEN, BRAF, and ras we show that both H-ras overexpression and expression of an oncogenic ras allele (ras V12) reduce susceptibility to lapatinib in analogy to what observed with activating PI3KCA mutations and with a constitutively active form of Akt. Importantly, we found that resistance to lapatinib due to ras overexpression or to ras V12 is overcome by MEK inhibition with U0126, suggesting a key role for the MEK-Erk pathway in ras-induced resistance. Similar results were obtained in BT474 cells, another HER+ breast cancer cell line. Therefore, our data indicate that overexpressed/mutated ras may act as a biological modifier of the response to lapatinib. Combining MEK inhibitors with lapatinib may help overcome this form of resistance and increase the efficacy of lapatinib in these tumors.
Current cancer drug targets 03/2010; 10(2):168-75. · 5.13 Impact Factor
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Computational Intelligence Methods for Bioinformatics and Biostatistics - 7th International Meeting, CIBB 2010, Palermo, Italy, September 16-18, 2010, Revised Selected Papers; 01/2010
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ABSTRACT: Monoclonal antibodies have been the most successful therapeutics ever brought to cancer treatment by immune technologies. The use of monoclonal antibodies in B-cell Non-Hodgkin's lymphomas (NHL) represents the greatest example of these advances, as the introduction of the anti-CD20 antibody rituximab has had a dramatic impact on how we treat this group of diseases today. Despite this success, several questions about how to optimize the use of monoclonal antibodies in NHL remain open. The best administration schedules, as well as the optimal duration of rituximab treatment, have yet to be determined. A deeper knowledge of the mechanisms underlying resistance to rituximab is also necessary in order to improve the activity of this and of similar therapeutics. Finally, new antibodies and biological agents are entering the scene and their advantages over rituximab will have to be assessed. We will discuss these issues and present an overview of the most significant clinical studies with monoclonal antibodies for NHL treatment carried out to date.
Clinical and Developmental Immunology 01/2010; 2010:428253. · 1.84 Impact Factor
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Alessio Nencioni,
Michele Cea,
Anna Garuti,
Mario Passalacqua,
Lizzia Raffaghello,
Debora Soncini, Eva Moran,
Gabriele Zoppoli,
Vito Pistoia,
Franco Patrone,
Alberto Ballestrero
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ABSTRACT: The efficacy of anti-HER2 therapeutics, such as lapatinib and trastuzumab, is limited by primary and acquired resistance. Cellular adaptations that allow breast cancer cell to survive prolonged HER2 inhibition include de-repression of the transcription factor FOXO3A with consequent estrogen receptor activation, and/or increased HER3 signaling. Here, we used low-density arrays, quantitative PCR, and western blotting to determine how HER2 signaling inhibition with lapatinib or PI3K inhibitors affects the expression of genes involved in breast cancer metastatic spread and overall prognosis. Retroviral transgenesis was used to express constitutively active forms of Akt in the HER2(+) breast cancer cell line SKBR3, and Grb7 in MCF7 cells. Specific gene silencing was obtained by siRNAs transfection. A murine BT474 xenograft cancer model was used to assess the effect of lapatinib on gene expression in vivo. We found that lapatinib induces upregulation of Grb7, an adaptor protein involved in receptor tyrosine kinase signaling and promoting cell survival and cell migration. Grb7 upregulation induced by lapatinib was found to occur in cancer cells in vitro and in vivo. We demonstrate that Grb7 upregulation is recreated by PI3K inhibitors while being prevented by constitutively active Akt. Thus, Grb7 is repressed by PI3K signaling and lapatinib-mediated Akt inhibition is responsible for Grb7 de-repression. Finally, we show that Grb7 removal by RNA-interference reduces breast cancer cell viability and increases the activity of lapatinib. In conclusion, Grb7 upregulation is a potentially adverse consequence of HER2 signaling inhibition. Preventing Grb7 accumulation and/or its interaction with receptor tyrosine kinases may increase the benefit of HER2-targeting drugs.
PLoS ONE 01/2010; 5(2):e9024. · 4.09 Impact Factor
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Michele Cea,
Gabriele Zoppoli,
Santina Bruzzone,
Floriana Fruscione, Eva Moran,
Anna Garuti,
Ilaria Rocco,
Gabriella Cirmena,
Salvatore Casciaro,
Francesca Olcese,
Ivana Pierri,
Antonia Cagnetta,
Fabio Ferrando,
Riccardo Ghio,
Marco Gobbi,
Alberto Ballestrero,
Franco Patrone,
Alessio Nencioni
Blood 07/2009; 113(23):6035-7; author reply 6037-8. · 9.90 Impact Factor
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Santina Bruzzone,
Floriana Fruscione,
Sara Morando,
Tiziana Ferrando,
Alessandro Poggi,
Anna Garuti,
Agustina D'Urso,
Martina Selmo,
Federica Benvenuto,
Michele Cea,
Gabriele Zoppoli, Eva Moran,
Debora Soncini,
Alberto Ballestrero,
Bernard Sordat,
Franco Patrone,
Raul Mostoslavsky,
Antonio Uccelli,
Alessio Nencioni
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ABSTRACT: Nicotinamide phosphoribosyltransferase (Nampt) inhibitors such as FK866 are potent inhibitors of NAD(+) synthesis that show promise for the treatment of different forms of cancer. Based on Nampt upregulation in activated T lymphocytes and on preliminary reports of lymphopenia in FK866 treated patients, we have investigated FK866 for its capacity to interfere with T lymphocyte function and survival. Intracellular pyridine nucleotides, ATP, mitochondrial function, viability, proliferation, activation markers and cytokine secretion were assessed in resting and in activated human T lymphocytes. In addition, we used experimental autoimmune encephalomyelitis (EAE) as a model of T-cell mediated autoimmune disease to assess FK866 efficacy in vivo. We show that activated, but not resting, T lymphocytes undergo massive NAD(+) depletion upon FK866-mediated Nampt inhibition. As a consequence, impaired proliferation, reduced IFN-gamma and TNF-alpha production, and finally autophagic cell demise result. We demonstrate that upregulation of the NAD(+)-degrading enzyme poly-(ADP-ribose)-polymerase (PARP) by activated T cells enhances their susceptibility to NAD(+) depletion. In addition, we relate defective IFN-gamma and TNF-alpha production in response to FK866 to impaired Sirt6 activity. Finally, we show that FK866 strikingly reduces the neurological damage and the clinical manifestations of EAE. In conclusion, Nampt inhibitors (and possibly Sirt6 inhibitors) could be used to modulate T cell-mediated immune responses and thereby be beneficial in immune-mediated disorders.
PLoS ONE 01/2009; 4(11):e7897. · 4.09 Impact Factor
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ABSTRACT: Dendritic cells are professional antigen-presenting cells with a key role in both immunity induction and tolerance maintenance. Dendritic cells are highly specialized in antigen capture, processing and presentation, and express co-stimulation signals which activate T lymphocytes and NK cells. Dendritic cells generated in culture and loaded with an antigen efficiently induce antigen-specific immunity after injection. More recently, methods have been developed that target antigens to dendritic cells in vivo, bypassing the need for ex vivo cell manipulations. Numerous ongoing studies aim to evaluate the effectiveness of dendritic cell vaccines in preventing tumor relapses and extending patients' survival. Further implementation of this form of immunotherapy is expected following the identification of the mechanisms controlling dendritic cell immunogenicity, and from a better understanding of the cell dynamics whereby immune responses are orchestrated. Here, we discuss these new insights together with an overview of the dendritic cell-based clinical studies carried out to date.
Advanced Drug Delivery Reviews 02/2008; 60(2):173-83. · 11.50 Impact Factor
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ABSTRACT: Proteasomal degradation is the main mechanism accounting for intracellular protein degradation. Not only is the proteasome involved in physiological protein turnover, it is also called into play by processes such as signal transduction, cell cycle, and apoptosis. Despite the ubiquitous distribution of the proteasome and its putative essential function, proteasome inhibitors have been developed that can be safely administered with acceptable side effects. Importantly, these compounds have been found to possess antitumor activity and are presently incorporated into the treatment of multiple myeloma and mantle cell lymphoma. In 2003, bortezomib (velcade) was the first compound in this category to have received FDA approval. The mechanisms for the antitumor activity of bortezomib and related drugs remain elusive. NF- kappaB inhibition by proteasome inhibitors may play a role in some instances. Recently, terminally differentiated plasma cells have been shown to downregulate proteasome expression and overall proteasome activity, which may account for the exquisite susceptibility of multiple myeloma cells to proteasome inhibition. New proteasome inhibitors with improved pharmacological properties are being developed and will soon enter clinical experimentation. Finally, studies addressing the usefulness of these compounds in other types of malignancies are ongoing and may soon expand the number of applications of these new therapeutic agents.
Journal of B.U.ON.: official journal of the Balkan Union of Oncology 10/2007; 12 Suppl 1:S95-9. · 0.61 Impact Factor
