95 results on '"Maria Chiara Fontana"'
Search Results
2. Inotuzumab ozogamicin is effective in relapsed/refractory extramedullary B acute lymphoblastic leukemia
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Luca Bertamini, Jacopo Nanni, Giovanni Marconi, Mariachiara Abbenante, Valentina Robustelli, Francesco Bacci, Antonella Matti, Stefania Paolini, Chiara Sartor, Silvia Lo Monaco, Maria Chiara Fontana, Stefano De Polo, Michele Cavo, Antonio Curti, Giovanni Martinelli, and Cristina Papayannidis
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Inotuzumab ozogamicin ,Extramedullary ,Acute lymphoblastic leukemia ,PET-CT scan ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Abstract Background Extramedullary involvement of B-cell Acute Lymphoblastic Leukemia (EM-ALL) is a rare occurrence, characterized by dismal outcome and the absence of a defined and shared therapeutic approach. In the landscape of innovative compounds, inotuzumab ozogamicin (IO) is a promising drug, whose mechanism of action relies on the killing of CD22 positive leukemic cells, through the delivery, after cell binding, of a molecule of calicheamicin. Case presentation We report two cases of CD22 positive relapsed EM-ALL treated with IO, obtained as compassionate use. Case 1, a 66 years old woman, affected by Philadelphia (Ph) negative B-ALL, relapsed with extramedullary involvement after 6 standard chemotherapy courses, who reached a complete metabolic response with IO treatment. Case 2, a 67 years old man with Ph positive B-ALL, initially treated with ponatinib, a third generation tyrosine-kinase inhibitor (TKI), obtaining a prolonged deep molecular remission. Nevertheless, for skin relapse during TKI treatment, the patient received local radiotherapy and, shortly after, standard chemotherapy, as multiple abdominal sites of relapse were detected too, with no response. The patient then received IO, obtained as compassionate use, with a good metabolic response. Conclusions These two cases suggest a possible key role of IO in the setting of advanced CD22 positive ALL, and underline its potential activity also in patients with EM involvement, relapsed after or refractory to conventional chemotherapy. Despite the well known hepatotoxic effect of the compound (Sinusoid Occlusive Syndrome), neither of them had such adverse event, moreover the second patient safely underwent allogeneic bone marrow transplantation.
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- 2018
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3. Targeting WEE1 to enhance conventional therapies for acute lymphoblastic leukemia
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Andrea Ghelli Luserna Di Rorà, Neil Beeharry, Enrica Imbrogno, Anna Ferrari, Valentina Robustelli, Simona Righi, Elena Sabattini, Maria Vittoria Verga Falzacappa, Chiara Ronchini, Nicoletta Testoni, Carmen Baldazzi, Cristina Papayannidis, Maria Chiara Abbenante, Giovanni Marconi, Stefania Paolini, Sarah Parisi, Chiara Sartor, Maria Chiara Fontana, Serena De Matteis, Ilaria Iacobucci, Pier Giuseppe Pelicci, Michele Cavo, Timothy J. Yen, and Giovanni Martinelli
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Acute lymphoblastic leukemia ,WEE1 inhibitor ,Chemo-sensitizer agent ,G2/M checkpoint ,Diseases of the blood and blood-forming organs ,RC633-647.5 ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Abstract Background Despite the recent progress that has been made in the understanding and treatment of acute lymphoblastic leukemia (ALL), the outcome is still dismal in adult ALL cases. Several studies in solid tumors identified high expression of WEE1 kinase as a poor prognostic factor and reported its role as a cancer-conserving oncogene that protects cancer cells from DNA damage. Therefore, the targeted inhibition of WEE1 kinase has emerged as a rational strategy to sensitize cancer cells to antineoplastic compounds, which we evaluate in this study. Methods The effectiveness of the selective WEE1 inhibitor AZD-1775 as a single agent and in combination with different antineoplastic agents in B and T cell precursor ALL (B/T-ALL) was evaluated in vitro and ex vivo studies. The efficacy of the compound in terms of cytotoxicity, induction of apoptosis, and changes in gene and protein expression was assessed using different B/T-ALL cell lines and confirmed in primary ALL blasts. Results We showed that WEE1 was highly expressed in adult primary ALL bone marrow and peripheral blood blasts (n = 58) compared to normal mononuclear cells isolated from the peripheral blood of healthy donors (p = 0.004). Thus, we hypothesized that WEE1 could be a rational target in ALL, and its inhibition could enhance the cytotoxicity of conventional therapies used for ALL. We evaluated the efficacy of AZD-1775 as a single agent and in combination with several antineoplastic agents, and we elucidated its mechanisms of action. AZD-1775 reduced cell viability in B/T-ALL cell lines by disrupting the G2/M checkpoint and inducing apoptosis. These findings were confirmed in human primary ALL bone marrow and peripheral blood blasts (n = 15). In both cell lines and primary leukemic cells, AZD-1775 significantly enhanced the efficacy of several tyrosine kinase inhibitors (TKIs) such as bosutinib, imatinib, and ponatinib, and of chemotherapeutic agents (clofarabine and doxorubicin) in terms of the reduction of cell viability, apoptosis induction, and inhibition of proliferation. Conclusions Our data suggest that WEE1 plays a role in ALL blast’s survival and is a bona fide target for therapeutic intervention. These data support the evaluation of the therapeutic potential of AZD-1775 as chemo-sensitizer agent for the treatment of B/T-ALL.
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- 2018
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4. Pharmacological Inhibition of WIP1 Sensitizes Acute Myeloid Leukemia Cells to the MDM2 Inhibitor Nutlin-3a
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Maria Chiara Fontana, Jacopo Nanni, Andrea Ghelli Luserna di Rorà, Elisabetta Petracci, Antonella Padella, Martina Ghetti, Anna Ferrari, Giovanni Marconi, Simona Soverini, Ilaria Iacobucci, Cristina Papayannidis, Antonio Curti, Ernesta Audisio, Maria Benedetta Giannini, Michela Rondoni, Francesco Lanza, Michele Cavo, Giovanni Martinelli, and Giorgia Simonetti
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AML ,novel therapeutic targets ,WIP1 ,MDM2 ,Biology (General) ,QH301-705.5 - Abstract
In acute myeloid leukemia (AML), the restoration of p53 activity through MDM2 inhibition proved efficacy in combinatorial therapies. WIP1, encoded from PPM1D, is a negative regulator of p53. We evaluated PPM1D expression and explored the therapeutic efficacy of WIP1 inhibitor (WIP1i) GSK2830371, in association with the MDM2 inhibitor Nutlin-3a (Nut-3a) in AML cell lines and primary samples. PPM1D transcript levels were higher in young patients compared with older ones and in core-binding-factor AML compared with other cytogenetic subgroups. In contrast, its expression was reduced in NPM1-mutated (mut, irrespective of FLT3-ITD status) or TP53-mut cases compared with wild-type (wt) ones. Either Nut-3a, and moderately WIP1i, as single agent decreased cell viability of TP53-wt cells (MV-4-11, MOLM-13, OCI-AML3) in a time/dosage-dependent manner, but not of TP53-mut cells (HEL, KASUMI-1, NOMO-1). The drug combination synergistically reduced viability and induced apoptosis in TP53-wt AML cell line and primary cells, but not in TP53-mut cells. Gene expression and immunoblotting analyses showed increased p53, MDM2 and p21 levels in treated TP53-wt cells and highlighted the enrichment of MYC, PI3K-AKT-mTOR and inflammation-related signatures upon WIP1i, Nut-3a and their combination, respectively, in the MV-4-11 TP53-wt model. This study demonstrated that WIP1 is a promising therapeutic target to enhance Nut-3a efficacy in TP53-wt AML.
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- 2021
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5. Acute Myeloid Leukemia Mutations: Therapeutic Implications
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Cristina Papayannidis, Chiara Sartor, Giovanni Marconi, Maria Chiara Fontana, Jacopo Nanni, Gianluca Cristiano, Sarah Parisi, Stefania Paolini, and Antonio Curti
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acute myeloid leukemia ,mutations ,FLT3 ,IDH1-2 ,resistance ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Acute Myeloid Leukemia (AML) is an extremely heterogeneous group of hematological neoplasms, for which allogeneic stem cell transplantation (HSCT) still represents the only potentially curative option in the majority of cases. However, elderly age and clinically severe comorbidities may often exclude a wide amount of patients from this therapeutic approach, underlying the urgent need for alternative strategies. Thanks to the introduction of advanced high-throughput techniques, light is being shed on the pathogenesis of AML, identifying molecular recurrent mutations as responsible for the onset, as well as progression, of disease. As a consequence, and in parallel, many new compounds, including targeted therapies (FMS-like tyrosine kinase 3 (FLT3) and Isocitrate dehydrogenase 1-2 (IDH1-2) inhibitors), have found a wide room of application in this setting, and are now available in daily practice, or in late phases of clinical development. Moreover, several further innovative molecules are currently under investigation, and promising results for many of them have already been reported. In this review, we will present an update on the most relevant molecular alterations of AML, focusing on the most frequent genomic mutations of the disease, for which compounds have been approved or are still currently under investigation.
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- 2019
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6. Data from Release of IFNγ by Acute Myeloid Leukemia Cells Remodels Bone Marrow Immune Microenvironment by Inducing Regulatory T Cells
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Antonio Curti, Marilena Ciciarello, Michele Cavo, Sergio Rutella, Mario Paolo Colombo, Giovanni Martinelli, Milena Piccioli, Darina Ocadlikova, Emanuela Ottaviani, Lorenza Bandini, Gianluca Cristiano, Claudio Tripodo, Alessandro Gulino, Martina Pazzaglia, Maria Chiara Fontana, Jayakumar Vadakekolathu, Sabina Sangaletti, Giorgia Simonetti, Barbara Bassani, and Giulia Corradi
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Purpose:The stromal and immune bone marrow (BM) landscape is emerging as a crucial determinant for acute myeloid leukemia (AML). Regulatory T cells (Treg) are enriched in the AML microenvironment, but the underlying mechanisms are poorly elucidated. Here, we addressed the effect of IFNγ released by AML cells in BM Treg induction and its impact on AML prognosis.Experimental Design:BM aspirates from patients with AML were subdivided according to IFNG expression. Gene expression profiles in INFγhigh and IFNγlow samples were compared by microarray and NanoString analysis and used to compute a prognostic index. The IFNγ release effect on the BM microenvironment was investigated in mesenchymal stromal cell (MSC)/AML cell cocultures. In mice, AML cells silenced for ifng expression were injected intrabone.Results:IFNγhigh AML samples showed an upregulation of inflammatory genes, usually correlated with a good prognosis in cancer. In contrast, in patients with AML, high IFNG expression was associated with poor overall survival. Notably, IFNγ release by AML cells positively correlated with a higher BM suppressive Treg frequency. In coculture experiments, IFNγhigh AML cells modified MSC transcriptome by upregulating IFNγ-dependent genes related to Treg induction, including indoleamine 2,3-dioxygenase 1 (IDO1). IDO1 inhibitor abrogated the effect of IFNγ release by AML cells on MSC-derived Treg induction. In vivo, the genetic ablation of IFNγ production by AML cells reduced MSC IDO1 expression and Treg infiltration, hindering AML engraftment.Conclusions:IFNγ release by AML cells induces an immune-regulatory program in MSCs and remodels BM immunologic landscape toward Treg induction, contributing to an immunotolerant microenvironment.See related commentary by Ferrell and Kordasti, p. 2986
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- 2023
7. Supplementary Data from Release of IFNγ by Acute Myeloid Leukemia Cells Remodels Bone Marrow Immune Microenvironment by Inducing Regulatory T Cells
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Antonio Curti, Marilena Ciciarello, Michele Cavo, Sergio Rutella, Mario Paolo Colombo, Giovanni Martinelli, Milena Piccioli, Darina Ocadlikova, Emanuela Ottaviani, Lorenza Bandini, Gianluca Cristiano, Claudio Tripodo, Alessandro Gulino, Martina Pazzaglia, Maria Chiara Fontana, Jayakumar Vadakekolathu, Sabina Sangaletti, Giorgia Simonetti, Barbara Bassani, and Giulia Corradi
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Supplementary Data from Release of IFNγ by Acute Myeloid Leukemia Cells Remodels Bone Marrow Immune Microenvironment by Inducing Regulatory T Cells
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- 2023
8. Release of IFNγ by Acute Myeloid Leukemia Cells Remodels Bone Marrow Immune Microenvironment by Inducing Regulatory T Cells
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Giulia Corradi, Barbara Bassani, Giorgia Simonetti, Sabina Sangaletti, Jayakumar Vadakekolathu, Maria Chiara Fontana, Martina Pazzaglia, Alessandro Gulino, Claudio Tripodo, Gianluca Cristiano, Lorenza Bandini, Emanuela Ottaviani, Darina Ocadlikova, Milena Piccioli, Giovanni Martinelli, Mario Paolo Colombo, Sergio Rutella, Michele Cavo, Marilena Ciciarello, Antonio Curti, Corradi, Giulia, Bassani, Barbara, Simonetti, Giorgia, Sangaletti, Sabina, Vadakekolathu, Jayakumar, Fontana, Maria Chiara, Pazzaglia, Martina, Gulino, Alessandro, Tripodo, Claudio, Cristiano, Gianluca, Bandini, Lorenza, Ottaviani, Emanuela, Ocadlikova, Darina, Piccioli, Milena, Martinelli, Giovanni, Colombo, Mario Paolo, Rutella, Sergio, Cavo, Michele, Ciciarello, Marilena, and Curti, Antonio
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Cancer Research ,Bone Marrow Cells ,Mesenchymal Stem Cells ,Settore MED/08 - Anatomia Patologica ,T-Lymphocytes, Regulatory ,Interferon-gamma ,Leukemia, Myeloid, Acute ,Mice ,Oncology ,Bone Marrow ,hemic and lymphatic diseases ,Tumor Microenvironment ,Animals ,IFNγ, Acute Myeloid Leukemia, Bone Marrow Immune Microenvironment - Abstract
Purpose: The stromal and immune bone marrow (BM) landscape is emerging as a crucial determinant for acute myeloid leukemia (AML). Regulatory T cells (Treg) are enriched in the AML microenvironment, but the underlying mechanisms are poorly elucidated. Here, we addressed the effect of IFNγ released by AML cells in BM Treg induction and its impact on AML prognosis. Experimental Design: BM aspirates from patients with AML were subdivided according to IFNG expression. Gene expression profiles in INFγhigh and IFNγlow samples were compared by microarray and NanoString analysis and used to compute a prognostic index. The IFNγ release effect on the BM microenvironment was investigated in mesenchymal stromal cell (MSC)/AML cell cocultures. In mice, AML cells silenced for ifng expression were injected intrabone. Results: IFNγhigh AML samples showed an upregulation of inflammatory genes, usually correlated with a good prognosis in cancer. In contrast, in patients with AML, high IFNG expression was associated with poor overall survival. Notably, IFNγ release by AML cells positively correlated with a higher BM suppressive Treg frequency. In coculture experiments, IFNγhigh AML cells modified MSC transcriptome by upregulating IFNγ-dependent genes related to Treg induction, including indoleamine 2,3-dioxygenase 1 (IDO1). IDO1 inhibitor abrogated the effect of IFNγ release by AML cells on MSC-derived Treg induction. In vivo, the genetic ablation of IFNγ production by AML cells reduced MSC IDO1 expression and Treg infiltration, hindering AML engraftment. Conclusions: IFNγ release by AML cells induces an immune-regulatory program in MSCs and remodels BM immunologic landscape toward Treg induction, contributing to an immunotolerant microenvironment. See related commentary by Ferrell and Kordasti, p. 2986
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- 2022
9. Genome-wide association study identifies susceptibility loci for acute myeloid leukemia
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Anne M. Dickinson, Gail Jones, David C. Linch, Clare Lendrem, David Grimwade, Richard A. Larson, Andrew D. Skol, Yaobo Xu, Adam Ivey, Wei-Yu Lin, Manja Meggendorfer, Rosemary E. Gale, Inés Gómez-Seguí, Giovani Marconi, Jean Norden, Jude Fitzgibbon, Mette K. Andersen, M Bornhäuser, Sarah E. Fordham, Amanda F. Gilkes, Heinz Sill, Eric A. Hungate, José Cervera, Friedrich Stölzel, Julia Gaal-Wesinger, Kim Piechocki, Wendy Stock, Theresa Hahn, Konstantin Strauch, David Allsup, Kenan Onel, Claire Elstob, Alyssa I. Clay-Gilmour, Nicola J. Sunter, Jelena D. Milosevic Feenstra, Meyling Cheok, Abrar Alharbi, Ann K. Daly, Sally Jeffries, Lisa Wagenführ, Olaf Heidenreich, Robert Kralovics, Alan K. Burnett, Giovanni Martinelli, Desiree Kunadt, Christian Gieger, Francesco Lo-Coco, Leo Ruhnke, Maria Teresa Voso, Junke Wang, Catherine Park, Nigel H. Russell, Chimène Moreilhon, Robert Kerrin Hills, Claude Preudhomme, Graham Jackson, Daniel Nowak, Maria Chiara Fontana, James M. Allan, Heidi Altmann, Richard S. Houlston, Anne S. Quante, Michelle M. Le Beau, Thahira Rahman, Christoph Röllig, Rebecca Darlay, Sophie Raynaud, Helen Marr, Csaba Bödör, Louise Palm, Thomas Cluzeau, Szilvia Krizsán, Heather J. Cordell, Mathew Collin, Torsten Haferlach, Lara E. Sucheston-Campbell, Wolf-Karsten Hofmann, Kimmo Porkka, Andras Masszi, Hervé Dombret, Miguel A. Sanz, Elisabeth Douglas, Tobias Menne, HUS Comprehensive Cancer Center, University Management, Helsinki University Hospital Area, Department of Oncology, and Hematologian yksikkö
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Oncology ,General Physics and Astronomy ,Genome-wide association study ,Disease ,0302 clinical medicine ,AML ,HLA Antigens ,hemic and lymphatic diseases ,Histone methylation ,Cancer genomics ,RISK ,0303 health sciences ,Multidisciplinary ,Myeloid leukemia ,Middle Aged ,CLONAL EVOLUTION ,CANCER ,3. Good health ,HLA ,Leukemia, Myeloid, Acute ,030220 oncology & carcinogenesis ,GENE-MUTATIONS ,medicine.medical_specialty ,Genotype ,Science ,Locus (genetics) ,HIF-1-ALPHA ,Human leukocyte antigen ,Polymorphism, Single Nucleotide ,Article ,Acute myeloid leukaemia ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Aldehyde Reductase ,Internal medicine ,medicine ,Humans ,Genetic Predisposition to Disease ,030304 developmental biology ,Genetic association ,OLDER PATIENTS ,Whites ,business.industry ,HUMAN-LEUKOCYTE ANTIGEN ,Reproducibility of Results ,Cancer ,General Chemistry ,Settore MED/15 ,medicine.disease ,IMMUNE ESCAPE ,Risk factors ,Case-Control Studies ,3111 Biomedicine ,business ,Genome-Wide Association Study - Abstract
Acute myeloid leukemia (AML) is a hematological malignancy with an undefined heritable risk. Here we perform a meta-analysis of three genome-wide association studies, with replication in a fourth study, incorporating a total of 4018 AML cases and 10488 controls. We identify a genome-wide significant risk locus for AML at 11q13.2 (rs4930561; P = 2.15 × 10−8; KMT5B). We also identify a genome-wide significant risk locus for the cytogenetically normal AML sub-group (N = 1287) at 6p21.32 (rs3916765; P = 1.51 × 10−10; HLA). Our results inform on AML etiology and identify putative functional genes operating in histone methylation (KMT5B) and immune function (HLA)., Genome wide association studies in cancer are used to understand the heritable genetic contribution to disease risk. Here, the authors perform a genome wide association study in European patients with acute myeloid leukemia and identify loci associated with risk of developing the disease.
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- 2021
10. Genome-wide association study identifies susceptibility loci for acute myeloid leukemia
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Michelle M. Le Beau, Thahira Rahman, Yaobo Xu, Wendy Stock, Andrew D. Skol, Abrar Alharbi, David Allsup, Claire Elstob, Lara E. Sucheston-Campbell, Lisa Wagenführ, Olaf Heidenreich, Claude Preudhomme, Tobias Menne, Szilvia Krizsán, Rebecca Darlay, Jelena D. Milosevic Feenstra, David C. Linch, Sophie Raynaud, Helen Marr, Christian Gieger, Francesco Lo-Coco, David Grimwade, Maria Teresa Voso, Junke Wang, Christoph Röllig, Clare Lendrem, Wolf-Karsten Hofmann, Mathew Collin, Manja Meggendorfer, Friedrich Stölzel, Wei-Yu Lin, Ann K. Daly, Theresa Hahn, Torsten Haferlach, Sally Jeffries, Julia Gaal-Wesinger, Konstantin Strauch, Giovani Marconi, Amanda F. Gilkes, Chimène Moreilhon, Giovanni Martinelli, Anne M. Dickinson, Robert Kerrin Hills, Alan K. Burnett, Mette K. Andersen, Leo Ruhnke, Kimmo Porkka, Catherine Park, Desiree Kunadt, Nigel H. Russell, M Bornhäuser, Alyssa I. Clay-Gilmour, Hervé Dombret, Sarah E. Fordham, Eric A. Hungate, Miguel A. Sanz, Inés Gómez-Seguí, Csaba Bödör, Jean Norden, Elisabeth Douglas, Rosemary E. Gale, Heinz Sill, Kim Piechocki, Richard A. Larson, Robert Kralovics, Meyling Cheok, Heidi Altmann, Richard S. Houlston, Andras Masszi, Anne S. Quante, Louise Palm, Thomas Cluzeau, Heather J. Cordell, Nicola J. Sunter, Graham Jackson, Daniel Nowak, Maria Chiara Fontana, James M. Allan, José Cervera, Kenan Onel, Gail Jones, Adam Ivey, and Jude Fitzgibbon
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0303 health sciences ,Myeloid leukemia ,Locus (genetics) ,Genome-wide association study ,Human leukocyte antigen ,Biology ,3. Good health ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,030220 oncology & carcinogenesis ,Histone methylation ,Cancer research ,Etiology ,030304 developmental biology ,Genetic association - Abstract
Acute myeloid leukemia (AML) is a hematological malignancy with an undefined heritable risk. Here we performed a meta-analysis of three genome-wide association studies, with replication in a fourth study, incorporating a total of 4018 AML cases and 10488 controls. We identified a genome-wide significant risk locus for AML at 11q13.2 (rs4930561; P = 2.15 × 10-8; KMT5B). We also identified a genome-wide significant risk locus for the cytogenetically normal AML sub-group (N=1287) at 6p21.32 (rs3916765; P = 1.51 × 10-10; HLA). Our results inform on AML etiology by identifying putative functional genes operating in histone methylation (KMT5B) and immune function (HLA).
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- 2021
11. Exploring the ATR-CHK1 Pathway in the Response of Doxorubicin-induced DNA Damages in Acute Lymphoblastic Leukemia Cells
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Claudio Cerchione, Antonella Padella, Anna Maria Ferrari, Andrea Ghelli Luserna di Rorà, Lorenzo Ledda, Enrica Imbrogno, Giorgia Simonetti, Monica Fumagalli, Maria Teresa Bochicchio, Maria Chiara Fontana, Roberta Napolitano, Annalisa Imovilli, Gerardo Musuraca, Martina Ghetti, Matteo Bocconcelli, Giovanni Martinelli, Chiara Liverani, Michela Rondoni, Matteo Paganelli, Seydou Sanogo, and Valentina Robustelli
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biology ,Chemistry ,Health, Toxicology and Mutagenesis ,Topoisomerase ,T cell ,Cell Biology ,Cell cycle ,Toxicology ,Prexasertib ,medicine.anatomical_structure ,Apoptosis ,biology.protein ,medicine ,Cancer research ,Cytotoxic T cell ,biological phenomena, cell phenomena, and immunity ,Fragmentation (cell biology) ,Mitosis - Abstract
Doxorubicin (Dox) is one of the most commonly used anthracyclines for the treatment of solid and hematological tumors such as B−/T cell acute lymphoblastic leukemia (ALL). Dox compromises topoisomerase II enzyme functionality, thus inducing structural damages during DNA replication and causes direct damages intercalating into DNA double helix. Eukaryotic cells respond to DNA damages by activating the ATM-CHK2 and/or ATR-CHK1 pathway, whose function is to regulate cell cycle progression, to promote damage repair, and to control apoptosis. We evaluated the efficacy of a new drug schedule combining Dox and specific ATR (VE-821) or CHK1 (prexasertib, PX) inhibitors in the treatment of human B−/T cell precursor ALL cell lines and primary ALL leukemic cells. We found that ALL cell lines respond to Dox activating the G2/M cell cycle checkpoint. Exposure of Dox-pretreated ALL cell lines to VE-821 or PX enhanced Dox cytotoxic effect. This phenomenon was associated with the abrogation of the G2/M cell cycle checkpoint with changes in the expression pCDK1 and cyclin B1, and cell entry in mitosis, followed by the induction of apoptosis. Indeed, the inhibition of the G2/M checkpoint led to a significant increment of normal and aberrant mitotic cells, including those showing tripolar spindles, metaphases with lagging chromosomes, and massive chromosomes fragmentation. In conclusion, we found that the ATR-CHK1 pathway is involved in the response to Dox-induced DNA damages and we demonstrated that our new in vitro drug schedule that combines Dox followed by ATR/CHK1 inhibitors can increase Dox cytotoxicity against ALL cells, while using lower drug doses. Graphical abstract • Doxorubicin activates the G2/M cell cycle checkpoint in acute lymphoblastic leukemia (ALL) cells. • ALL cells respond to doxorubicin-induced DNA damages by activating the ATR-CHK1 pathway. • The inhibition of the ATR-CHK1 pathway synergizes with doxorubicin in the induction of cytotoxicity in ALL cells. • The inhibition of ATR-CHK1 pathway induces aberrant chromosome segregation and mitotic spindle defects in doxorubicin-pretreated ALL cells.
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- 2021
12. Integrated genomic-metabolic classification of acute myeloid leukemia defines a subgroup with NPM1 and cohesin/DNA damage mutations
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Samantha Bruno, Andrea Ghelli Luserna di Rorà, Anna Maria Ferrari, Giovanni Marconi, Rossella De Tommaso, Carlo Mengucci, Maria Teresa Bochicchio, Cristina Papayannidis, Margherita Perricone, Antonella Padella, Francesco Capozzi, Claudio Delpino, Torsten Haferlach, Jacopo Nanni, Emanuela Ottaviani, Emanuela Scarpi, Martina Pazzaglia, Gastone Castellani, Eugenia Franchini, Giovanni Martinelli, Gianfranco Picone, Martina Ghetti, Annalisa Astolfi, Maria Chiara Fontana, Ilaria Iacobucci, Giorgia Simonetti, Michele Cavo, Roberta Napolitano, Viviana Guadagnuolo, Michela Tebaldi, Eugenio Fonzi, Daniel Remondini, Carmen Baldazzi, Simonetti, Giorgia, Mengucci, Carlo, Padella, Antonella, Fonzi, Eugenio, Picone, Gianfranco, Delpino, Claudio, Nanni, Jacopo, De Tommaso, Rossella, Franchini, Eugenia, Papayannidis, Cristina, Marconi, Giovanni, Pazzaglia, Martina, Perricone, Margherita, Scarpi, Emanuela, Fontana, Maria Chiara, Bruno, Samantha, Tebaldi, Michela, Ferrari, Anna, Bochicchio, Maria Teresa, Ghelli Luserna Di Rorà, Andrea, Ghetti, Martina, Napolitano, Roberta, Astolfi, Annalisa, Baldazzi, Carmen, Guadagnuolo, Viviana, Ottaviani, Emanuela, Iacobucci, Ilaria, Cavo, Michele, Castellani, Gastone, Haferlach, Torsten, Remondini, Daniel, Capozzi, Francesco, and Martinelli, Giovanni
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0301 basic medicine ,Male ,Cancer Research ,Chromosomal Proteins, Non-Histone ,Metabolic alteration ,Cell Cycle Proteins ,medicine.disease_cause ,0302 clinical medicine ,hemic and lymphatic diseases ,Cancer genomics ,Genomic subgroup ,Purine metabolism ,Aged, 80 and over ,Mutation ,Myeloid leukemia ,Nuclear Proteins ,Hematology ,Genomics ,Middle Aged ,Prognosis ,Cancer metabolism ,Chromatin ,3. Good health ,Metabolic cluster ,Leukemia, Myeloid, Acute ,Oncology ,030220 oncology & carcinogenesis ,Molecular classification ,Female ,Nucleophosmin ,Adult ,NPM1 ,congenital, hereditary, and neonatal diseases and abnormalities ,Adolescent ,DNA damage ,Metabolomic ,Biology ,Article ,Acute myeloid leukaemia ,03 medical and health sciences ,Young Adult ,Metabolomics ,medicine ,Metabolome ,Humans ,Aged ,030104 developmental biology ,Cancer research ,DNA Damage - Abstract
Although targeting of cell metabolism is a promising therapeutic strategy in acute myeloid leukemia (AML), metabolic dependencies are largely unexplored. We aimed to classify AML patients based on their metabolic landscape and map connections between metabolic and genomic profiles. Combined serum and urine metabolomics improved AML characterization compared with individual biofluid analysis. At intracellular level, AML displayed dysregulated amino acid, nucleotide, lipid, and bioenergetic metabolism. The integration of intracellular and biofluid metabolomics provided a map of alterations in the metabolism of polyamine, purine, keton bodies and polyunsaturated fatty acids and tricarboxylic acid cycle. The intracellular metabolome distinguished three AML clusters, correlating with distinct genomic profiles: NPM1-mutated(mut), chromatin/spliceosome-mut and TP53-mut/aneuploid AML that were confirmed by biofluid analysis. Interestingly, integrated genomic-metabolic profiles defined two subgroups of NPM1-mut AML. One was enriched for mutations in cohesin/DNA damage-related genes (NPM1/cohesin-mut AML) and showed increased serum choline + trimethylamine-N-oxide and leucine, higher mutation load, transcriptomic signatures of reduced inflammatory status and better ex-vivo response to EGFR and MET inhibition. The transcriptional differences of enzyme-encoding genes between NPM1/cohesin-mut and NPM1-mut allowed in silico modeling of intracellular metabolic perturbations. This approach predicted alterations in NAD and purine metabolism in NPM1/cohesin-mut AML that suggest potential vulnerabilities, worthy of being therapeutically explored.
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- 2021
13. Loss of PALB2 predicts poor prognosis in acute myeloid leukemia and suggests novel therapeutic strategies targeting the DNA repair pathway
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Andrea Ghelli Luserna di Rorà, Maria Chiara Fontana, Giorgia Simonetti, Giovanni Marconi, Cristina Papayannidis, Antonella Padella, Nicoletta Testoni, Giovanni Martinelli, Torsten Haferlach, Anna Maria Ferrari, Gastone Castellani, Elisabetta Petracci, Eugenio Fonzi, Carmen Baldazzi, Padella A., Fontana M.C., Marconi G., Fonzi E., Petracci E., Ferrari A., Baldazzi C., Papayannidis C., Ghelli Luserna Di Rora A., Testoni N., Castellani G., Haferlach T., Martinelli G., and Simonetti G.
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Adult ,Male ,Poor prognosis ,DNA Repair ,PALB2 ,Synthetic lethality ,Polymorphism, Single Nucleotide ,lcsh:RC254-282 ,Acute myeloid leukaemia ,03 medical and health sciences ,Young Adult ,0302 clinical medicine ,Text mining ,hemic and lymphatic diseases ,Correspondence ,Cancer genomics ,Medicine ,Humans ,030304 developmental biology ,Aged ,Aged, 80 and over ,0303 health sciences ,business.industry ,Myeloid leukemia ,Hematology ,DNA Repair Pathway ,Middle Aged ,Prognosis ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,synthetic lethality ,3. Good health ,Leukemia, Myeloid, Acute ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,Female ,business ,Fanconi Anemia Complementation Group N Protein ,Gene Deletion - Abstract
Dear Editor, Acute myeloid leukemia (AML) patients carrying complex karyotype or aneuploidies have a very poor prognosis, with a 5-year overall survival (OS)
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- 2021
14. Safety profile and impact on survival of tyrosine kinase inhibitors versus conventional therapy in relapse or refractory FLT3 positive acute myeloid leukemia patients
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Maria Teresa Bochicchio, Michele Cavo, Sarah Parisi, Carmen Baldazzi, Chiara Sartor, Giovanni Martinelli, Jacopo Nanni, Simone Ragaini, Matteo Olivi, Nicoletta Testoni, Stefano De Polo, Antonio Curti, Maddalena Raffini, Maria Chiara Fontana, Cristina Papayannidis, Emanuela Ottaviani, Francesca Bonifazi, Annalisa Talami, Gianluca Cristiano, Stefania Paolini, Mariachiara Abbenante, Giovanni Marconi, Luca Bertamini, Marconi G., De Polo S., Martinelli G., Nanni J., Bertamini L., Talami A., Olivi M., Ragaini S., Abbenante M.C., Sartor C., Ottaviani E., Bochicchio M.T., Parisi S., Fontana M.C., Cristiano G., Raffini M., Baldazzi C., Testoni N., Bonifazi F., Paolini S., Curti A., Cavo M., and Papayannidis C.
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Oncology ,Adult ,Male ,Cancer Research ,medicine.medical_specialty ,Survival ,Adolescent ,medicine.medical_treatment ,Antineoplastic Agents ,Disease ,Disease-Free Survival ,03 medical and health sciences ,0302 clinical medicine ,Refractory ,AML ,Internal medicine ,hemic and lymphatic diseases ,Medicine ,Chemotherapy ,Humans ,Adverse effect ,FLT3 ,Protein Kinase Inhibitors ,Aged ,business.industry ,Surrogate endpoint ,Myeloid leukemia ,Retrospective cohort study ,Hematology ,Middle Aged ,TKI ,respiratory tract diseases ,Survival Rate ,Leukemia, Myeloid, Acute ,fms-Like Tyrosine Kinase 3 ,030220 oncology & carcinogenesis ,Mutation ,Quality of Life ,Neoplastic cell ,Female ,Safety ,business ,030215 immunology - Abstract
Relapsed or refractory (R/R) acute myeloid leukemia (AML) has a poor prognosis, and new therapies are a major clinical need. When mutated, FLT3 drives neoplastic cell proliferation. New drugs (i.e., tyrosine kinase inhibitors, TKIs) showed effectiveness in FLT3-AML and promise to change disease history and outcome. We evaluated the benefit conferred by TKIs in terms of survival, burden of complications and surrogate endpoint of quality of life in a retrospective cohort of 49 FLT3 positive, R/R AML patients. Patients who received TKIs were compared to those treated with conventional chemotherapy. Treatment with TKIs conferred a better OS and wea associated with a lower burden and severity of adverse events. Importantly, patients who received TKIs showed reduced time of hospitalization. In conclusion, treatment with TKI in R/R FLT3-AML was related to a better survival, less and milder AEs, and shorter hospitalization.
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- 2020
15. Targeting WEE1 to enhance conventional therapies for acute lymphoblastic leukemia
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Simona Righi, Valentina Robustelli, Sarah Parisi, Nicoletta Testoni, Maria Vittoria Verga Falzacappa, Chiara Sartor, Cristina Papayannidis, Maria Chiara Abbenante, Timothy J. Yen, Michele Cavo, Andrea Ghelli Luserna di Rorà, Chiara Ronchini, Serena De Matteis, Pier Giuseppe Pelicci, Enrica Imbrogno, Anna Maria Ferrari, Neil Beeharry, Giovanni Marconi, Maria Chiara Fontana, Giovanni Martinelli, Ilaria Iacobucci, Carmen Baldazzi, Elena Sabattini, Stefania Paolini, Ghelli Luserna Di Rorà, Andrea, Beeharry, Neil, Imbrogno, Enrica, Ferrari, Anna, Robustelli, Valentina, Righi, Simona, Sabattini, Elena, Verga Falzacappa, Maria Vittoria, Ronchini, Chiara, Testoni, Nicoletta, Baldazzi, Carmen, Papayannidis, Cristina, Abbenante, Maria Chiara, Marconi, Giovanni, Paolini, Stefania, Parisi, Sarah, Sartor, Chiara, Fontana, Maria Chiara, De Matteis, Serena, Iacobucci, Ilaria, Pelicci, Pier Giuseppe, Cavo, Michele, Yen, Timothy J, and Martinelli, Giovanni
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0301 basic medicine ,Cancer Research ,medicine.medical_specialty ,T cell ,Cell Cycle Proteins ,Acute lymphoblastic leukemia ,Peripheral blood mononuclear cell ,lcsh:RC254-282 ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cell Line, Tumor ,Internal medicine ,medicine ,Humans ,Clofarabine ,Molecular Biology ,G2/M checkpoint ,Hematology ,business.industry ,lcsh:RC633-647.5 ,Research ,Ponatinib ,Nuclear Proteins ,lcsh:Diseases of the blood and blood-forming organs ,Precursor Cell Lymphoblastic Leukemia-Lymphoma ,Protein-Tyrosine Kinases ,Chemo-sensitizer agent ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,3. Good health ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,chemistry ,030220 oncology & carcinogenesis ,Cancer cell ,Cancer research ,WEE1 inhibitor ,business ,Bosutinib ,Tyrosine kinase ,medicine.drug - Abstract
Background Despite the recent progress that has been made in the understanding and treatment of acute lymphoblastic leukemia (ALL), the outcome is still dismal in adult ALL cases. Several studies in solid tumors identified high expression of WEE1 kinase as a poor prognostic factor and reported its role as a cancer-conserving oncogene that protects cancer cells from DNA damage. Therefore, the targeted inhibition of WEE1 kinase has emerged as a rational strategy to sensitize cancer cells to antineoplastic compounds, which we evaluate in this study. Methods The effectiveness of the selective WEE1 inhibitor AZD-1775 as a single agent and in combination with different antineoplastic agents in B and T cell precursor ALL (B/T-ALL) was evaluated in vitro and ex vivo studies. The efficacy of the compound in terms of cytotoxicity, induction of apoptosis, and changes in gene and protein expression was assessed using different B/T-ALL cell lines and confirmed in primary ALL blasts. Results We showed that WEE1 was highly expressed in adult primary ALL bone marrow and peripheral blood blasts (n = 58) compared to normal mononuclear cells isolated from the peripheral blood of healthy donors (p = 0.004). Thus, we hypothesized that WEE1 could be a rational target in ALL, and its inhibition could enhance the cytotoxicity of conventional therapies used for ALL. We evaluated the efficacy of AZD-1775 as a single agent and in combination with several antineoplastic agents, and we elucidated its mechanisms of action. AZD-1775 reduced cell viability in B/T-ALL cell lines by disrupting the G2/M checkpoint and inducing apoptosis. These findings were confirmed in human primary ALL bone marrow and peripheral blood blasts (n = 15). In both cell lines and primary leukemic cells, AZD-1775 significantly enhanced the efficacy of several tyrosine kinase inhibitors (TKIs) such as bosutinib, imatinib, and ponatinib, and of chemotherapeutic agents (clofarabine and doxorubicin) in terms of the reduction of cell viability, apoptosis induction, and inhibition of proliferation. Conclusions Our data suggest that WEE1 plays a role in ALL blast’s survival and is a bona fide target for therapeutic intervention. These data support the evaluation of the therapeutic potential of AZD-1775 as chemo-sensitizer agent for the treatment of B/T-ALL. Electronic supplementary material The online version of this article (10.1186/s13045-018-0641-1) contains supplementary material, which is available to authorized users.
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- 2018
16. Impact of Comorbidities on Prognosis of Elderly Patients with Acute Myeloid Leukemia Who Receive Hypomethylating Agents
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Sarah Parisi, Chiara Sartor, Renato Fanin, Davide Lazzarotto, Stefania Paolini, Maria Chiara Abbenante, Giovanni Martinelli, Maria Chiara Fontana, Nicoletta Testoni, Michele Cavo, Gianluca Cristiano, Maria Benedetta Giannini, Rania Abd-alatif, Cristina Papayannidis, Anna Candoni, Chiara Di Giovanni Bezzi, Antonio Curti, Carmen Baldazzi, Emanuela Ottaviani, Jacopo Nanni, Giovanni Marconi, Roberta di Nicola, and Lorenza Bandini
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Oncology ,medicine.medical_specialty ,business.industry ,Internal medicine ,Immunology ,medicine ,Myeloid leukemia ,Cell Biology ,Hematology ,business ,Biochemistry - Abstract
BACKGROUND: Many efforts have been made in the attempt to address the conundrum question of fitness definition ad prognosis prediction in elderly acute myeloid leukemia (AML) patients. Parametric definitions are expected to give an advantage in patient stratification; however, clinical examination remain de facto pivotal to formulate therapy decisions and frequently the comorbidities are empirically evaluated. METHODS: We conducted a multicenter study collecting baseline comorbidity, laboratory data, CTCAE 4.0.3 adverse events (AE), and outcome of elderly patients (>65 years old) with new onset AML who received hypomethylating agents as 1st line therapy. We tested the impact on prognosis of baseline clinical and biological risk factors. Furthermore, we evaluated a score - acute myeloid leukemia-composite model, AML-CM (Mukherjee et al, 2017) - developed in chemotherapy-eligible patients, that accounts for baseline comorbidities, laboratory parameters, age and cytogenetic-molecular risk. The study was approved by local Ethical Authority (316/2019/Oss/AOUBo). RESULTS: We collected data from 131 consecutive elderly patients who received 1 st line HMAs between January 2008 and January 2021. Patients had a median age of 76 years (IQR 72 -79). Seventy-seven out of 131 patients (58.8%) had de novo AML, 32/131 (32.8%) had secondary AML, and 11/131 (8.4%) had therapy-related AML. Out of 123 evaluable patients, 43 (34.9%) had complex karyotype, 1 (0.8%) inv(16), 59 (48.4) normal karyotype, 18 (14.7%) other alterations; 8/108 patients harbored FLT3 ITD mutation (7.4%, 23 not tested), 12/101 NPM1 mutation (11.9%, 30 not tested). Based on these data, 111 patients were evaluable for ELN2010 risk stratification; 9 over 111 patients (8.1%) were stratified in the low risk, 42/111 (37.8%) in intermediate-1 risk, 17/111(15.3%) in intermediate-2 risk, and 43/111 (38.7%) in high-risk class. As expected, most of the patients had at least one comorbidity. Particularly, baseline arrhythmia was present in 29/130 (22.1%, 1 no data), cardiovascular comorbidity in 20/130 (15.4%, 1 no data), diabetes in 20/131 (15.3%), cerebrovascular comorbidity in 11/131 (8.4%), kidney disease in 15/130 (11.5%, 1 no data), lung chronic disease 19/130 (14.6% 1 no data), hypoalbuminemia in 25/111 patients (22.5%, 20 no data). With a median follow up of 28.2 months, median overall survival (OS) of the entire cohort was 15.8 months (95% C.I. 11.2-19.4). We confirmed that patient who obtained a response (complete remission, partial response, hematological improvement) after 2 months of therapy had the best OS (figure A, median OS of 21 months for responders vs 7.4 months for non-responders, p To test the impact of comorbidities combined with cytogenetic and molecular risk, AML-CM was used. Our results indicate that AML-CM score was able to stratify prognosis in elderly patients receiving frontline HMAs (figure B, median OS in score group 1: 29.7 months, score group 2: 16.5 months, score group 3: 11.2 months, score group 4: 6.6 months, p=.038). The worse prognosis of patients with higher AML-CM score, which includes patients with increased baseline comorbidities, may be explained with a higher incidence of AEs (84.55, 116.01, 131.45, 229.3 events for 100 patients per year for score group 1,2,3, and 4, respectively) and infections (53.80, 55.10, 85.95, 140.13 events for 100 patients per year for score group 1,2,3, and 4, respectively), in patients with higher baseline comorbidities. CONCLUSION: In this study we found that baseline comorbidities, captured by AML-CM score, may define prognosis of elderly patients receiving 1st line HMAs; parametric comorbidity scores may improve our ability to predict outcome and tailor interventions. The impact of comorbidity on OS may be increased with novel and more aggressive therapy. For this reason, specific studies on functional fitness tests and geriatric assessments are highly warranted in patients receiving HMAs plus venetoclax. This work was supported by Bologna AIL. CP and AC shared last authorship. Figure 1 Figure 1. Disclosures Martinelli: Daichii Sankyo: Consultancy; Pfizer: Consultancy, Speakers Bureau; Astellas: Consultancy, Speakers Bureau; Roche: Consultancy; Abbvie: Consultancy; Stemline Therapeutics: Consultancy; Celgene /BMS: Consultancy, Speakers Bureau; Incyte: Consultancy; Jazz Pharmaceuticals: Consultancy. Cavo: Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel Accommodations, Speakers Bureau; Amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Takeda: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Novartis: Honoraria; AbbVie: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: TRAVEL, ACCOMMODATIONS, EXPENSES, Speakers Bureau; Adaptive Biotechnologies: Consultancy, Honoraria; GlaxoSmithKline: Consultancy, Honoraria; Sanofi: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Bristol-Myers Squib: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau. Papayannidis: Pfizer, Amgen, Novartis: Honoraria. Curti: Abbvie: Membership on an entity's Board of Directors or advisory committees; Novartis: Membership on an entity's Board of Directors or advisory committees; Pfizer: Membership on an entity's Board of Directors or advisory committees; Jazz Pharma: Membership on an entity's Board of Directors or advisory committees.
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- 2021
17. Aneuploid acute myeloid leukemia exhibits a signature of genomic alterations in the cell cycle and protein degradation machinery
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Elisa Ficarra, Ilaria Iacobucci, Carmen Baldazzi, Giorgia Simonetti, Elisa Zuffa, Emanuela Ottaviani, Jesús M. Hernández, Antonella Padella, Stefania Paolini, Eugenia Franchini, Federica Zanotti, Giovanni Martinelli, Peter Vandenberghe, Anna Maria Ferrari, Samantha Bruno, Marco Sazzini, Torsten Haferlach, Nicoletta Testoni, Gastone Castellani, Marco Manfrini, Annalisa Astolfi, Simona Bernardi, Italo Faria do Valle, Maria Antonella Laginestra, Jan Cools, Maria Chiara Fontana, Cristina Papayannidis, Giovanni Marconi, Eugenio Fonzi, Daniel Remondini, Michele Cavo, Viviana Guadagnuolo, Lars Bullinger, Simonetti, Giorgia, Padella, Antonella, do Valle, Italo Farìa, Fontana, Maria Chiara, Fonzi, Eugenio, Bruno, Samantha, Baldazzi, Carmen, Guadagnuolo, Viviana, Manfrini, Marco, Ferrari, Anna, Paolini, Stefania, Papayannidis, Cristina, Marconi, Giovanni, Franchini, Eugenia, Zuffa, Elisa, Laginestra, Maria Antonella, Zanotti, Federica, Astolfi, Annalisa, Iacobucci, Ilaria, Bernardi, Simona, Sazzini, Marco, Ficarra, Elisa, Hernandez, Jesus Maria, Vandenberghe, Peter, Cools, Jan, Bullinger, Lar, Ottaviani, Emanuela, Testoni, Nicoletta, Cavo, Michele, Haferlach, Torsten, Castellani, Gastone, Remondini, Daniel, and Martinelli, Giovanni
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Male ,Cancer Research ,Hematologic Malignancies ,Gene Dosage ,Aneuploidy ,medicine.disease_cause ,whole exome sequencing ,genomic ,acute myeloid leukemia, aneuploidy, cell cycle, genomics, mutation, ubiquitination, whole exome sequencing ,0302 clinical medicine ,hemic and lymphatic diseases ,Gene Regulatory Networks ,030212 general & internal medicine ,Aged, 80 and over ,Mutation ,Gene Expression Regulation, Leukemic ,Myeloid leukemia ,bioinformatics ,Middle Aged ,Cell cycle ,3. Good health ,Leukemia, Myeloid, Acute ,Oncology ,030220 oncology & carcinogenesis ,Female ,Original Article ,cell cycle ,Erratum ,Adult ,acute myeloid leukemia ,aneuploidy ,genomics ,mutation ,ubiquitination ,Cancer Research, bioinformatics ,DNA repair ,Protein degradation ,NO ,Young Adult ,03 medical and health sciences ,Exome Sequencing ,medicine ,Humans ,Genetic Predisposition to Disease ,Aged ,business.industry ,Gene Expression Profiling ,Original Articles ,medicine.disease ,Protein ubiquitination ,Chromosome Banding ,Rad50 ,Proteolysis ,Cancer research ,Disease Site ,business - Abstract
Background Aneuploidy occurs in more than 20% of acute myeloid leukemia (AML) cases and correlates with an adverse prognosis. Methods To understand the molecular bases of aneuploid acute myeloid leukemia (A‐AML), this study examined the genomic profile in 42 A‐AML cases and 35 euploid acute myeloid leukemia (E‐AML) cases. Results A‐AML was characterized by increased genomic complexity based on exonic variants (an average of 26 somatic mutations per sample vs 15 for E‐AML). The integration of exome, copy number, and gene expression data revealed alterations in genes involved in DNA repair (eg, SLX4IP, RINT1, HINT1, and ATR) and the cell cycle (eg, MCM2, MCM4, MCM5, MCM7, MCM8, MCM10, UBE2C, USP37, CK2, CK3, CK4, BUB1B, NUSAP1, and E2F) in A‐AML, which was associated with a 3‐gene signature defined by PLK1 and CDC20 upregulation and RAD50 downregulation and with structural or functional silencing of the p53 transcriptional program. Moreover, A‐AML was enriched for alterations in the protein ubiquitination and degradation pathway (eg, increased levels of UHRF1 and UBE2C and decreased UBA3 expression), response to reactive oxygen species, energy metabolism, and biosynthetic processes, which may help in facing the unbalanced protein load. E‐AML was associated with BCOR/BCORL1 mutations and HOX gene overexpression. Conclusions These findings indicate that aneuploidy‐related and leukemia‐specific alterations cooperate to tolerate an abnormal chromosome number in AML, and they point to the mitotic and protein degradation machineries as potential therapeutic targets., Aneuploid acute myeloid leukemia (A‐AML) is associated with genomic and transcriptional alterations in the cell cycle and protein degradation pathways. The upregulation of PLK1 and CDC20 and the downregulation of RAD50 and of a p53‐related signature are hallmarks of A‐AML.
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- 2019
18. Novel and Rare Fusion Transcripts Involving Transcription Factors and Tumor Suppressor Genes in Acute Myeloid Leukemia
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Simona Soverini, Viviana Guadagnuolo, Elisa Ficarra, Giulia Paciello, Andrea Ghelli Luserna di Rorà, Clelia Tiziana Storlazzi, Antonella Padella, Maria Chiara Fontana, Carmen Baldazzi, Nicoletta Testoni, Anna Maria Ferrari, Rossella De Tommaso, Simona Righi, Cristina Papayannidis, Valentina Robustelli, George Giotopoulos, Eugenia Franchini, Samantha Bruno, Giovanni Martinelli, Claudia Haferlach, Martina Ghetti, Emanuela Ottaviani, Brian J. P. Huntly, Anna Stengel, Elena Sabattini, Giorgia Simonetti, Ilaria Iacobucci, Apollo - University of Cambridge Repository, Giotopoulos, George [0000-0003-1390-6592], Huntly, Brian [0000-0003-0312-161X], Padella A., Simonetti G., Paciello G., Giotopoulos G., Baldazzi C., Righi S., Ghetti M., Stengel A., Guadagnuolo V., De Tommaso R., Papayannidis C., Robustelli V., Franchini E., Di Rora A.G.L., Ferrari A., Fontana M.C., Bruno S., Ottaviani E., Soverini S., Storlazzi C.T., Haferlach C., Sabattini E., Testoni N., Iacobucci I., Huntly B.J.P., Ficarra E., and Martinelli G.
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0301 basic medicine ,Cancer Research ,BCL11B ,Biology ,ZEB2-BCL11B ,acute myeloid leukemia ,Article ,03 medical and health sciences ,Chimera (genetics) ,0302 clinical medicine ,hemic and lymphatic diseases ,medicine ,Transcription factor ,medicine.diagnostic_test ,rare fusion genes ,Myeloid leukemia ,medicine.disease ,Phenotype ,Leukemia ,030104 developmental biology ,Oncology ,Fusion transcript ,030220 oncology & carcinogenesis ,Cancer research ,Rare fusion gene ,Fluorescence in situ hybridization - Abstract
Approximately 18% of acute myeloid leukemia (AML) cases express a fusion transcript. However, few fusions are recurrent across AML and the identification of these rare chimeras is of interest to characterize AML patients. Here, we studied the transcriptome of 8 adult AML patients with poorly described chromosomal translocation(s), with the aim of identifying novel and rare fusion transcripts. We integrated RNA-sequencing data with multiple approaches including computational analysis, Sanger sequencing, fluorescence in situ hybridization and in vitro studies to assess the oncogenic potential of the ZEB2-BCL11B chimera. We detected 7 different fusions with partner genes involving transcription factors (OAZ-MAFK, ZEB2-BCL11B), tumor suppressors (SAV1-GYPB, PUF60-TYW1, CNOT2-WT1) and rearrangements associated with the loss of NF1 (CPD-PXT1, UTP6-CRLF3). Notably, ZEB2-BCL11B rearrangements co-occurred with FLT3 mutations and were associated with a poorly differentiated or mixed phenotype leukemia. Although the fusion alone did not transform murine c-Kit+ bone marrow cells, 45.4% of 14q32 non-rearranged AML cases were also BCL11B-positive, suggesting a more general and complex mechanism of leukemogenesis associated with BCL11B expression. Overall, by combining different approaches, we described rare fusion events contributing to the complexity of AML and we linked the expression of some chimeras to genomic alterations hitting known genes in AML.
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- 2019
19. Identification of Two DNMT3A Mutations Compromising Protein Stability and Methylation Capacity in Acute Myeloid Leukemia
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Maddalena Raffini, Samantha Bruno, Maria Chiara Fontana, Martina Pazzaglia, Anna Maria Ferrari, Valentina Robustelli, Simona Soverini, Andrea Ghelli Luserna di Rorà, Maria Teresa Bochicchio, Claudia Venturi, Giovanna Prisinzano, Cristina Papayannidis, Lorenza Bandini, Emanuela Ottaviani, Giovanni Marconi, Eugenia Franchini, Chiara Sartor, Giovanni Martinelli, Maria Chiara Abbenante, Antonella Padella, Giorgia Simonetti, Bruno S., Bochicchio M.T., Franchini E., Padella A., Marconi G., Ghelli Luserna Di Rora A., Venturi C., Raffini M., Prisinzano G., Ferrari A., Bandini L., Robustelli V., Pazzaglia M., Fontana M.C., Sartor C., Abbenante M.C., Papayannidis C., Soverini S., Ottaviani E., Simonetti G., and Martinelli G.
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Acute Myeloid Leukemia ,0301 basic medicine ,Genetics ,Article Subject ,business.industry ,Myeloid leukemia ,Methylation ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,lcsh:RC254-282 ,Stop codon ,3. Good health ,Frameshift mutation ,03 medical and health sciences ,Exon ,030104 developmental biology ,0302 clinical medicine ,Oncology ,CpG site ,030220 oncology & carcinogenesis ,DNA methylation ,embryonic structures ,Missense mutation ,Medicine ,business ,Research Article - Abstract
Somatic mutations of DNMT3A occur in about 20% of acute myeloid leukemia (AML) patients. They mostly consist in heterozygous missense mutations targeting a hotspot site at R882 codon, which exhibit a dominant negative effect and are associated with high myeloblast count, advanced age, and poor prognosis. Other types of mutations such as truncations, insertions, or single-nucleotide deletion also affect the DNMT3A gene, though with lower frequency. The present study aimed to characterize two DNMT3A gene mutations identified by next-generation sequencing (NGS), through analysis of protein stability and DNA methylation status at CpG islands. The first mutation was a single-nucleotide variant of DNMT3A at exon 20 causing a premature STOP codon (c.2385G > A; p.Trp795∗; NM_022552.4). The DNMT3A mutation load increased from 4.5% to 38.2% during guadecitabine treatment, with a dominant negative effect on CpG methylation and on protein expression. The second mutation was a novel insertion of 35 nucleotides in exon 22 of DNMT3A (NM_022552.4) that introduced a STOP codon too, after the amino acid Glu863 caused by a frameshift insertion (c.2586_2587insTCATGAATGAGAAAGAGGACATCTTATGGTGCACT; p. Thr862_Glu863fsins). The mutation, which was associated with reduced DNMT3A expression and CpG methylation, persisted at relapse with minor changes in the methylation profile and at protein level. Our data highlight the need to better understand the consequences of DNMT3A mutations other than R882 substitutions in the leukemogenic process in order to tailor patient treatments, thus avoiding therapeutic resistance and disease relapse.
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- 2019
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20. The alteration in key regulator genes of autophagy is mainstream mechanism of therapy resistance and impact prognosis of acute myelogenous leukemia (AML): results from diagnosis genomic analysis on 148 consecutive patients treated with intensive chemotherapy and long-term survival follow-up
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Giovanni Marconi, Cristina Papayannidis, Maria Chiara Fontana, Antonella Padella, Anna Ferrari, Eugenia Franchini, Stefania Paolini, Maria Chiara Abbenante, Chiara Sartor, Francesca Volpato, Viviana Guadagnuolo, Silvia Lo Monaco, Elena Tenti, Andrea Ghelli Luserna Di Rora, Valentina Robustelli, Nicoletta Testoni, Giorgia Simonetti, Emanuela Ottaviani, Giovanni Martinelli, and Giovanni Marconi, Cristina Papayannidis, Maria Chiara Fontana, Antonella Padella, Anna Ferrari, Eugenia Franchini, Stefania Paolini, Maria Chiara Abbenante, Chiara Sartor, Francesca Volpato, Viviana Guadagnuolo, Silvia Lo Monaco, Elena Tenti, Andrea Ghelli Luserna Di Rora, Valentina Robustelli, Nicoletta Testoni, Giorgia Simonetti, Emanuela Ottaviani, Giovanni Martinelli
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Autophagy, Acute Myeloid Leukemia - Published
- 2017
21. Co-occurrence of alterations in the DNA damage repair genes synergize with uncontrolled proliferation and associate with very-poor prognosis in acute myeloid leukemia patients
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Antonella Padella, Giorgia Simonetti, Maria Chiara Fontana, Marco Manfrini, Giovanni Marconi, Anna Ferrari, Italo Faria do Valle, Marianna Garonzi, Cristina Papayannidis, Eugenia Franchini, Elisa Zuffa, Viviana Guadagnuolo, Samantha Bruno, Andrea Ghelli Luserna di Rorà, Emanuela Ottaviani, Daniel Remondini, Massimo Delledonne, Giovanni Martinelli, and Antonella Padella, Giorgia Simonetti, Maria Chiara Fontana, Marco Manfrini, Giovanni Marconi, Anna Ferrari, Italo Faria do Valle, Marianna Garonzi, Cristina Papayannidis, Eugenia Franchini, Elisa Zuffa, Viviana Guadagnuolo, Samantha Bruno, Andrea Ghelli Luserna di Rorà, Emanuela Ottaviani, Daniel Remondini, Massimo Delledonne, Giovanni Martinelli
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AML, DNA repair - Published
- 2017
22. Separase overexpression defines a new subset of acute myeloma leukemia patients characterized by high CD34 and MYC levels
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Giorgia Simonetti, Antonella Padella, Simona Righi, Maria Chiara Fontana, Marco Manfrini, Cristina Papayannidis, Giovanni Marconi, Carmen Baldazzi, Marianna Garonzi, Alberto Ferrarini, Massimo Delledonne, Nicoletta Testoni, Elena Sabattini, Giovanni Martinelli, and Giorgia Simonetti, Antonella Padella, Simona Righi, Maria Chiara Fontana, Marco Manfrini, Cristina Papayannidis, Giovanni Marconi, Carmen Baldazzi, Marianna Garonzi, Alberto Ferrarini, Massimo Delledonne, Nicoletta Testoni, Elena Sabattini, Giovanni Martinelli
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Acute Myeloid Leukemia, gene expression - Published
- 2017
23. Distinct pattern of alterations in tp53 mutated and wild type acute myeloid leukemia (AML) patients
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Anna Ferrari, Eugenio Fonzi, Maria Chiara Fontana, Andrea Ghelli Luserna Di Rorà, Marco Manfrini, Antonella Padella, Carmen Baldazzi, Cristina Papayannidis, Giovanni Marconi, Stefania Paolini, Viviana Guadagnuolo, Margherita Perricone, Valentina Robustelli, Enrica Imbrogno, Eugenia Franchini, Claudia Venturi, Elisa Zuffa, Maria Chiara Abbenante, Giorgia Simonetti, Nicoletta Testoni, Emanuela Ottaviani, Martinelli Giovanni, and Anna Ferrari, Eugenio Fonzi, Maria Chiara Fontana, Andrea Ghelli Luserna Di Rorà, Marco Manfrini, Antonella Padella, Carmen Baldazzi, Cristina Papayannidis, Giovanni Marconi, Stefania Paolini, Viviana Guadagnuolo, Margherita Perricone, Valentina Robustelli, Enrica Imbrogno, Eugenia Franchini, Claudia Venturi, Elisa Zuffa, Maria Chiara Abbenante, Giorgia Simonetti, Nicoletta Testoni, Emanuela Ottaviani, Martinelli Giovanni
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TP53, Acute Myeloid Leukemia - Published
- 2017
24. SETD2 Non-Genomic Loss of Function and p53 Functional Inactivation in Advanced Systemic Mastocytosis (SM): Pathogenetic and Therapeutic Implications
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Manuela Mancini, Cecilia Monaldi, Sara De Santis, Cristina Papayannidis, Michela Rondoni, Maria Chiara Abbenante, Luana Bavaro, Margherita Martelli, Elisa Ficarra, Giulia Paciello, Viviana Guadagnuolo, Maria Chiara Fontana, Marco Manfrini, Roberta Zanotti, Livio Pagano, Francesco Albano, Fabio Ciceri, Chiara Elena, Patrizia Tosi, Massimo Delledonne, Peter Valent, Michele Cavo, Giovanni Martinelli, Simona Soverini, and Manuela Mancini, Cecilia Monaldi, Sara De Santis, Cristina Papayannidis, Michela Rondoni, Maria Chiara Abbenante, Luana Bavaro, Margherita Martelli, Elisa Ficarra, Giulia Paciello, Viviana Guadagnuolo, Maria Chiara Fontana, Marco Manfrini, Roberta Zanotti, Livio Pagano, Francesco Albano, Fabio Ciceri, Chiara Elena, Patrizia Tosi, Massimo Delledonne, Peter Valent, Michele Cavo, Giovanni Martinelli, Simona Soverini
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Systemic Mastocytosis, SETD2 - Published
- 2017
25. In Systemic Masocytosis, Midostaurin Targets Both Kit and Aurora Kinase a Reverting H3K36Me3 Deficiency and Synergizes with Second-Generation Tyrosine Kinase Inhibitors
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Roberta Zanotti, Peter Valent, Massimiliano Bonifacio, Livio Pagano, Marianna Criscuolo, Chiara Elena, Patrizia Tosi, Cristina Papayannidis, Fabio Ciceri, Michela Rondoni, Samantha Bruno, Luana Bavaro, Luciano Xumerle, Margherita Martelli, Simona Soverini, Francesco Albano, Luigi Scaffidi, Giovanni Martinelli, Antonio Curti, Manuela Mancini, Michele Cavo, Maria Chiara Fontana, Sara De Santis, Cecilia Monaldi, Massimo Delledonne, and C. Avanzato
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business.industry ,Immunology ,Cell Biology ,Hematology ,medicine.disease ,Biochemistry ,Dasatinib ,chemistry.chemical_compound ,Leukemia ,chemistry ,Nilotinib ,medicine ,Cancer research ,Midostaurin ,Danusertib ,Aurora Kinase A ,Annexin A5 ,business ,Tyrosine kinase ,medicine.drug - Abstract
H3K36 tri-methylation by SETD2 has been linked to a plethora of pathways critical for the regulation of a multitude of cellular processes, including transcription, DNA replication and DNA repair after damage, yet the functional implications of perturbed H3K36 tri-methylation by SETD2 mutations or non-genomic loss of function in leukemia are still unclear. We have previously reported that the HMC-1.1 and -1.2 mast cell leukemia (MCL) cell lines and many patients (pts) with advanced systemic mastocytosis (advSM) display H3K36Me3 deficiency as a result of non-genomic loss of function of SETD2. Proteasome inhibition restored SETD2 protein expression and H3K36me3, suggesting that a functional protein is produced but rapidly degraded. To understand the mechanisms underlying this phenomenon, we used an in silico approach to identify candidate SETD2-interacting proteins, followed by experimental confirmation by co-immunoprecipitation. We found that, after proteasomal inhibition, SETD2 co-immunoprecipitates with Aurora Kinase A (AKA). We also found that AKA was overexpressed and hyper-activated in pts with advSM compared to ISM pts and to a pool of healthy donors, and that AKA can phosphorylate SETD2. Both pharmacological inhibition by Danusertib and siRNA-mediated knock-down of AKA rescued SETD2 expression and activity, raising the hypothesis that phosphorylation by AKA might be implicated in proteasome-mediated degradation of SETD2. The new standard of therapy in advSM is midostaurin, that inhibits the activity of both wild‐type and D816V mutant KIT, as well as of various other kinases including AKA. Therefore we investigated if midostaurin effects may be addressed to AKA inhibition and consequent SETD2/H3K36me3 rescue. To this purpose, HMC-1 cells were treated with 5 µM midostaurin for 24 h and AKA, SETD2 and H3K36me3 expression were evaluated by western blotting. Treatment with midostaurin was able to inhibit AKA activity by about 60%, partially restoring SETD2 expression and H3K36Me3. Moreover, midostaurin treatment of HMC-1 cells at micromolar doses induced cytostatic but not cytotoxic effects as shown by cell growth curves performed in liquid medium and as confirmed by annexin V/PI staining and subsequent cytofluorimetric analysis of apoptotic cell death. Our observations in cell lines were confirmed in neoplastic mast cells collected from six patients with advSM before and after three months of midostaurin treatment. Western blotting showed that midostaurin treatment in vivo indeed results in a rescue of SEDT2 expression and activity, associated with a partial de-phosphorylation of AKA. Our subsequent experimental step was therefore to hypothesize and test the possibility of a combined treatment between midostaurin and second generation TKIs to induce not only a cytostatic but also a cytotoxic effect both in our in vitro models and in primary cells obtained from bone marrow samples of advSM patients. We performed growth curves in liquid medium and clonogenic assays to evaluate the therapeutic potential of pharmacological combination of midostaurin with Nilotinib and Dasatinib in HMC-1 cells and in neoplastic mast cells from 3 patients with advSM and we observed in all cases synergistic effects at nanomolar doses. Moreover, cytofluorimetric analysis of apoptotic cell death in HMC-1 cells showed an important advantage in using the combination of the two drugs, compared to single agents, underlined by the significant reduction of drug doses used to obtain cytotoxic effects (Figure 1). Our results suggest that AKA-mediated post-translational modifications contribute to SETD2 non-genomic loss of function in advSM. Inhibiting AKA and c-Kit activity by midostaurin in combination with a second generation TKI is a promising therapeutic strategy in patients with SETD2/H3K36Me3 deficiency. Acknowledgments: study supported by AIRC (project code 16996) and AIL (Associazione Italiana contro le Leucemia, Linfomi e Mieloma). Disclosures Papayannidis: Incyte: Honoraria; Shire: Honoraria; Novartis: Honoraria; Teva: Honoraria; Amgen: Honoraria; Pfizer: Honoraria. Bonifacio:Pfizer: Honoraria; Amgen: Honoraria; Novartis: Honoraria; Incyte: Honoraria; BMS: Honoraria. Albano:Incyte: Membership on an entity's Board of Directors or advisory committees; Novartis: Membership on an entity's Board of Directors or advisory committees. Elena:Novartis: Consultancy; Pfizer: Consultancy. Valent:Deciphera: Honoraria, Research Funding; Blueprint: Research Funding; Pfizer: Honoraria; Celgene: Honoraria; Novartis: Consultancy, Honoraria, Research Funding. Cavo:celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: travel accommodations, Speakers Bureau; janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: travel accommodations, Speakers Bureau; bms: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; sanofi: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; novartis: Honoraria; takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; AbbVie: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees. Soverini:Incyte: Consultancy.
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- 2019
26. AML-CM Score Predicts Prognosis in Hemato-Geriatric Patients with New-Onset Acute Myeloid Leukemia (AML) Who Receive Hypomethylating Agents (HMA)
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Roberta di Nicola, Giovanni Marconi, Gianluca Cristiano, Stefania Paolini, Michele Baccarani, Carmen Baldazzi, Antonio Curti, Sarah Parisi, Lorenza Bandini, Maria Chiara Abbenante, Nicoletta Testoni, Maria Chiara Fontana, Jacopo Nanni, Chiara Sartor, Giovanni Martinelli, Cristina Papayannidis, Emanuela Ottaviani, and Michele Cavo
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medicine.medical_specialty ,education.field_of_study ,Incidence (epidemiology) ,Immunology ,Population ,Cell Biology ,Hematology ,medicine.disease ,Biochemistry ,Comorbidity ,Log-rank test ,symbols.namesake ,Interquartile range ,Internal medicine ,Chi-square test ,medicine ,symbols ,education ,Survival analysis ,Fisher's exact test - Abstract
Background Although much efforts have been made to precisely define fitness of AML patients, in patients who are not candidate to chemotherapy, there is no prognostic model and the respective weight of AML biology and patient fitness are not well established. Here we test AML-CM score (Sorror, JAMA 2018), that is validated in fit population, in a set of old AML patients who received HMAs. Methods We retrospectively collected data of consecutive patients who received HMAs in our institution from 1st Jan 2008 with an age > 65 years at AML diagnosis. AML-CM score was applied to all the patients. Patients were divided in 4 groups (score 1-4: group 1, score 5-6: group 2; score 7-9: group 3, score > 9: group 4) and in 2 macro-groups (score 1-6: group A and score > 6 group B) for the analyses. Descriptive data are presented as median with interquartile ranges (IQR). Adverse events are graded according to CTCAE v4.03. Survival analysis was conducted with Kaplan-Meyer and are presented as 95% confidence intervals (C.I.) and differences in overall survival (OS) were tested with 2-side log rank test. Fisher exact test and Person's chi squared test were used whenever appropriate. Results At data cut-off, 1st Jan 2019, 60 consecutive patients received decitabine or azacytidine as 1st line therapy for AML. Median age of the population was 75.94 years (IQR 72.53-80.38). Most of the patients (37/62, 59.7%) had de novo AML, 19/62 (30.6%) had AML secondary to previous myeloid disorders and 6/62 (9.7%) had AML secondary to chemotherapy or radiotherapy. Most of the patients were smokers (19/33, 57.57%, 29 no data), and few were usual drinkers (4/16, 25.00%, 46 no data). In our set, out of 62 patients, 2 patients (3.2%) had inv(3), 1 (1.6%) a translocation involving 11q23, 1 (1.6%) del(5q), 4 (6.4%) mon(7) or del (7q), 1 (1.6%) del(17p), 15 (24.2%) complex karyotype, 27 (43.5%) normal karyotype, 4 (6.5%) other alterations and 5 were not evaluable; 3/17 (17.65%, 45 no data) harbored IDH2 mutation, 1/16 (6.25%) IDH2 mutation, 2/33 FLT3 mutation (6.06%, 29 no data), 1/24 (4.17%, 38 no data), 2/15 (13.33%, 47 no data) TP53 mutation. According to ELN 2017, 3/62 patients (4.83%) had low risk, 34/62 (54.84%) intermediate risk and 23/62 (37.10%) high risk AML. According to AML-CM score, 13/62 patients (20.97%) were in group A, 20/62 (32.36%) in group B, 21/62 (33.87%) in group C, 6/62 (9.68%) in group D, 2/62 (3.23%) were not allocated for incomplete AML-CM score. There was no difference in term of age, ELN risk, secondary AML prevalence, HMA administered, or response to HMA according to ELN criteria between group 1, 2, 3, 4 or between macro-group A and B. Cardiovascular comorbidity, diabetes mellitus, obesity, previous tumor, hypoalbuminemia, elevated LDH were prevalent in higher risk AML-CM groups (3-4) and in macro-group B. Median OS was 658 days (95% C.I. 316-1000) in group 1, 556 days (95% C.I. 463-649 in group 2, 243 days (95% C.I. 153-353) in group 3, 107 days (95% C.I. 47-167) in group 4 (p=.021, figure 1A). Furthermore, we observed a median OS of 589 days (95% C.I. 328-850) in macro-group A and 219 days (95% C.I. 96-342) in macro-group B (p=.003, figure 1B). Reduced survival was correlated with a non-statistical trend toward augmented incidence of infections and adverse events in higher risk AML-CM groups (3-4). Conclusions AML-CM is a useful indicator of prognosis in old patients that receive HMAs. Prognosis in our set is influenced by comorbidity (measured with AML-CM, a quantitative score) more than by disease biology. We identified a group of patients (macro-group A) that has median OS after HMAs outlying OS reported in literature. This brilliant result can be due to lower comorbidity. AML-CM could help in defining candidate patients for therapy intensification and care utilization or for team comorbidity management. GM and RDN equally contributed Figure 1 Disclosures Martinelli: Roche: Consultancy; Novartis: Consultancy; ARIAD: Consultancy; BMS: Consultancy; Pfizer: Consultancy. Baccarani:Novartis: Consultancy, Speakers Bureau; Incyte: Consultancy, Speakers Bureau; Takeda: Consultancy. Papayannidis:Pfizer: Honoraria; Teva: Honoraria; Shire: Honoraria; Novartis: Honoraria; Amgen: Honoraria; Incyte: Honoraria. Cavo:janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: travel accommodations, Speakers Bureau; bms: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; sanofi: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: travel accommodations, Speakers Bureau; AbbVie: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; novartis: Honoraria; takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau.
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- 2019
27. Aurora Kinase a/MDM2-Mediated SETD2 Loss of Function in Chronic Myeloid Leukemia Patients in Blast Crisis Can be Therapeutically Targeted Inducing Apoptotic Cell Death in a Caspase-Dependent Way
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Elena Trabacchi, Francesco Albano, Giovanni Martinelli, Elisa Dan, Marzia Salvucci, Cecilia Monaldi, Elisabetta Calistri, Nicola Orofino, Luana Bavaro, Margherita Martelli, Massimiliano Bonifacio, Mario Tiribelli, Gabriele Gugliotta, Michele Cavo, Maria Chiara Fontana, Samantha Bruno, Sara Galimberti, Barbara Sinigaglia, Simona Soverini, Annalisa Imovilli, Fausto Castagnetti, Gianni Binotto, Michele Baccarani, Antonella Gozzini, Patrizia Pregno, Elena Tenti, Monica Crugnola, Alessandra Iurlo, Manuela Mancini, Gianantonio Rosti, Sara De Santis, Elisabetta Abruzzese, and Claudia Baratè
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biology ,business.industry ,education ,Immunology ,Myeloid leukemia ,Caspase 3 ,Cell Biology ,Hematology ,medicine.disease ,Biochemistry ,Leukemia ,Imatinib mesylate ,medicine ,biology.protein ,Cancer research ,Danusertib ,Aurora Kinase A ,business ,health care economics and organizations ,Caspase ,K562 cells - Abstract
One of the hallmarks of chronic myeloid leukemia (CML) is genomic instability, that fosters the acquisition of tyrosine kinase inhibitor (TKI)-resistant BCR-ABL1 mutations and/or of additional chromosomal aberrations leading to progression to blast crisis (BC). Inactivating mutations in the SETD2 tumor suppressor occur in solid tumors and acute leukemias. SETD2 trimethylates histone H3 Lysine 36 (H3K36Me3) playing a key role in maintaining DNA integrity. We have recently demonstrated that, in CML, SETD2 loss of function may occur at the post-translational level. Reduced or null SETD2 and H3K36Me3 was detected in 83/96 (86%) patients (pts) with BC CML as compared to a pool of healthy donors and to chronic phase (CP) pts at diagnosis. Proteasome inhibition in primary cells from pts with undetectable SETD2 restored H3K36Me3 and led to accumulation of hyper-ubiquitinated SETD2. In K562 cells (SETD2/H3K36Me3low), we observed that after proteasome inhibition hyper-ubiquitinated SETD2 co-immunoprecipitates with MDM2. MDM2 inhibition rescued SETD2 expression and activity, suggesting that MDM2 is implicated in SETD2 reduced stability. Co-IP also showed that SETD2 interacts with Aurora Kinase A (AKA) a S/T kinase frequently overexpressed in CML. We found that AKA phosphorylates SETD2, and its inhibition rescued SETD2 expression and activity. To investigate whether SETD2/H3K36Me3 loss may be a druggable lesion, we performed clonogenic assays in LAMA84 (SETD2/H3K36Me3high) cells before and after SETD2 silencing, in imatinib-sensitive K562 (SETD2/H3K36Me3low) cells and in IM-resistant K562 cells, that are characterized by complete SETD2 loss. The extent of reduction of clonogenic growth after proteasomal, AKA or MDM2 inhibition was found to be inversely correlated to SETD2 residual expression. These observations were confirmed in cells from both CP (n=2) and BC (n=4) CML pts showing different levels of SETD2 expression and activity. Further experiments were performed in the aforementioned cell lines to verify if reduced clonogenic potential was due to cytostatic or cytotoxic effects. Apoptotic cell death was quantified by annexin V/propidium iodide staining and flow cytometry. Proteasomal inhibition by bortezomib, carfilzomib and ixazomib and AKA de-phosphorylation by Danusertib caused a time-dependent increase of annexin-V-positive cells by activating the mitochondrial apoptotic pathway as reflected by an increase in Bax expression and induction of the cleavage of caspase-3,-9 and PARP. Moreover, all drug treatments as single agent, at nanomolar doses (Bortezomib: 10 nM, Carfilzomib: 5 nM, Ixazomib: 10 nM and Danusertib: 500 nM) induced a significant increase of the DNA double-strand break marker γH2AX, suggesting that in a SETD2 knock-down context, proteasomal and AKA inhibition propagates genomic instability by forcing the cells through successive replication cycles, ultimately resulting in apoptosis from mitotic catastrophe. Reduced SETD2/H3K36Me3 levels, in association with MDM2 and AKA hyper-activation, were also detected when the CD34+ cell fraction of 10 CML-CP pts, was compared to the total mononuclear cell fraction or to the CD34+ compartment obtained from a pool of healthy donors. We thus hypothesized that leukemia progenitor cells, showing higher MDM2 and AKA activity and consequent SETD2 loss, accumulate genetic aberrations despite inhibition of BCR-ABL1 kinase. Studies are ongoing to verify if MDM2 or AKA inhibition may restore SETD2 expression and function and induce cell death. Finally, it has already been shown that alterations of epigenetic regulators such as the KDM4 family members control tumor cell proliferation in a variety of cancers including acute myeloid leukemia. Recent findings have identified KDM4 demethylases as putative therapeutic targets in a SETD2 mutated context and illustrated the efficacy of KDM4 inhibitors in AML therapy. Starting from these evidences, we will test the same approach in BC CML models. In conclusion, phosphorylation by AKA and ubiquitination by MDM2 contribute to SETD2 non-genomic loss of function in BC CML and in CD34+ leukemic progenitors. Restoring physiological H3K36Me3 may help to improve the outcome of this critical subset of pts. Acknowledgments: Study supported by AIRC (project code 16996), AIL (Associazione Italiana contro le Leucemia, Linfomi e Mieloma), Italian Ministry of Health, project GR-2016-02364880. Disclosures Gugliotta: Pfizer: Honoraria; Novartis: Honoraria; Incyte: Honoraria. Castagnetti:Novartis: Honoraria; Incyte: Honoraria; Pfizer: Honoraria; Bristol Myers Squiib: Consultancy, Honoraria. Rosti:BMS: Speakers Bureau; Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Pfizer: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Incyte: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau. Iurlo:Incyte: Other: Speaker Honoraria; Novartis: Other: Speaker Honoraria; Pfizer: Other: Speaker Honoraria. Abruzzese:Incyte: Consultancy; Novartis: Consultancy; Pfizer: Consultancy; BMS: Consultancy. Pregno:Incyte: Consultancy, Honoraria; Pfizer: Honoraria; Novartis: Honoraria; Bristol Myers Squibb: Honoraria. Crugnola:Novartis: Honoraria; Incyte: Honoraria. Albano:Novartis: Membership on an entity's Board of Directors or advisory committees; Incyte: Membership on an entity's Board of Directors or advisory committees. Bonifacio:Novartis: Honoraria; Amgen: Honoraria; Pfizer: Honoraria; Incyte: Honoraria; BMS: Honoraria. Tiribelli:Pfizer: Membership on an entity's Board of Directors or advisory committees; Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Incyte: Membership on an entity's Board of Directors or advisory committees. Baccarani:Novartis: Consultancy, Speakers Bureau; Incyte: Consultancy, Speakers Bureau; Takeda: Consultancy. Martinelli:Roche: Consultancy; Pfizer: Consultancy; BMS: Consultancy; Novartis: Consultancy; ARIAD: Consultancy. Cavo:amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; AbbVie: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: travel accommodations, Speakers Bureau; novartis: Honoraria; takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; bms: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; sanofi: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: travel accommodations, Speakers Bureau. Soverini:Incyte: Consultancy.
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- 2019
28. The Prolonged Inhibition of Chk1/Chk2 Kinases Enhances Genetic Instability and Compromises the Efficacy of Chemotherapy Against Acute Lymphoblastic Leukemia Cells
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Antonella Padella, Martina Ghetti, Maria Chiara Fontana, Eugenio Fonzi, Anna Maria Ferrari, Roberta Napolitano, Maria Teresa Bochicchio, Giovanni Martinelli, Andrea Ghelli Luserna di Rorà, Enrica Imbrogno, and Giorgia Simonetti
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POLD1 ,Kinase ,DNA damage ,Immunology ,Cell Biology ,Hematology ,DNA Repair Pathway ,Biology ,medicine.disease ,Biochemistry ,Apoptosis ,Cell culture ,Acute lymphocytic leukemia ,Cancer cell ,medicine ,Cancer research - Abstract
Acute lymphoblastic leukemia (ALL) cells respond to chemotherapy, or more generally to DNA damages, by activating different DNA Damage Response (DDR) pathways. DDR-pathways regulate cell cycle progression and DNA damages repair. Molecular and functional alterations in key DDR-related genes drastically affect the effectiveness of DNA-damaging treatments in cancer cells. For this reasons selective DDR-inhibitors have been developed in order to sensitize cancer cells against conventional chemotherapy. Despite the proven efficacy of DDR-inhibitors in cancer treatment, only few studies have highlighted the biological consequences the prolonged inhibition of DDR-pathways in cancer cells. We hypothesized that the protracted inhibition of the DDR pathways may generate resistant clones characterized by an increased genetic instability. The aim of the study was to evaluate biological consequences of the prolonged inhibition of two crucial DDR-related kinase such as cell cycle checkpoint kinase 1 (Chk1) and 2 (Chk2) in B cells ALL. In particular, we investigated the consequences of Chk1/Chk2 inhibition in term of increase of genetic instability and in term of responsiveness to chemotherapy agents. Starting from B-ALL cell line NALM-6, we generated a resistant model (hereafter referred as N6R-PF8) by treating the parental cells with increasing concentration of PF-00477736 (Chk1/Chk2 inhibitor) for more than a year and increasing the IC50 value of 10-folds. From a molecular point of view, the N6R-PF8 accumulated significant molecular alterations. SNP microarray analysis highlighted different alterations in DDR-related genes and, in particular, in the ATM/CHK2 pathway. Three regions in copy number LOSS (CN=1) containing several genes involved in cell cycle checkpoint regulation (ATM and NPAT) and in the apoptosis (BIRC2, CASP1 and CASP5) were detectable only in NALM-6 parental cell lines and were copy number neutral in the resistant model. Immunoblotting analysis confirmed that in N6R-PF8 cells the ATM/CHK2 and ATR/CHK1 down-stream pathways were significant over-expressed and activated in comparison to the parental cell. Whole exome sequencing analysis showed that the two cell lines were characterized by different mutational profiles and that N6R-PF8 cells harboured significantly more genetic alterations in comparison with NALM-6 cells. Interestingly, crucial genes involved in DNA repair pathway (MLH3, NBN, POLD1 and PMS2) have been found altered only in N6R-PF8 cells. From a functional point of view, the molecular alterations characterizing the N6R-PF8 significantly compromised the cytotoxicity of PF-00477736 and of different DNA damaging agents in comparison to parental cells. Furthermore, the treatment with ATR/ATM inhibitor restored the sensitivity of N6R-PF8 to PF-00477736 and to different chemotherapy agents. In this scenario the level of expression of these two kinases seems to correlate with the sensitivity to DNA damaging agents and to PF-00477736. Finally, we confirmed that the protracted inhibition of crucial DDR-related kinase may increase the overall genetic instability in ALL cells and compromise the efficacy of DNA damaging based therapies. Disclosures Martinelli: Roche: Consultancy; ARIAD: Consultancy; Novartis: Consultancy; Pfizer: Consultancy; BMS: Consultancy.
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- 2019
29. SETD2 and histone H3 lysine 36 methylation deficiency in advanced systemic mastocytosis
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T Haferlach, Livio Pagano, Massimo Delledonne, Simona Soverini, Antonella Padella, Fabio Ciceri, Patrizia Tosi, L Riccioni, Marco Manfrini, Giovanni Martinelli, Manja Meggendorfer, Serena Merante, F Morigi, C De Benedittis, Domenica Gangemi, Luana Bavaro, Michela Rondoni, Manuela Mancini, Roberta Zanotti, Paolo Savini, Viviana Guadagnuolo, Giovanni Poletti, Michele Cavo, Maria Chiara Fontana, Luigi Scaffidi, Chiara Elena, Giorgina Specchia, Francesco Albano, Elisa Zago, Peter Valent, Raffaele A. Calogero, Cristina Papayannidis, Martinelli, G., Mancini, M., De Benedittis, C., Rondoni, M., Papayannidis, C., Manfrini, M., Meggendorfer, M., Calogero, R., Guadagnuolo, V., Fontana, M. C., Bavaro, L., Padella, A., Zago, E., Pagano, L., Zanotti, R., Scaffidi, L., Specchia, G., Albano, F., Merante, S., Elena, C., Savini, P., Gangemi, D., Tosi, P., Ciceri, F., Poletti, G., Riccioni, L., Morigi, F., Delledonne, M., Haferlach, T., Cavo, M., Valent, P., Soverini, S., Martinelli, G, Mancini, M, De Benedittis, C, Rondoni, M, Papayannidis, C, Manfrini, M, Meggendorfer, M, Calogero, R, Guadagnuolo, V, Fontana, M. C, Bavaro, L, Padella, A, Zago, E, Pagano, L, Zanotti, R, Scaffidi, L, Specchia, G, Albano, F, Merante, S, Elena, C, Savini, P, Gangemi, D, Tosi, P, Ciceri, F, Poletti, G, Riccioni, L, Morigi, F, Delledonne, M, Haferlach, T, Cavo, M, and Valent, P
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0301 basic medicine ,Adult ,Male ,Cancer Research ,Proteasome Endopeptidase Complex ,systemic mastocytosis, mast cell leukemia, SETD2 ,Apoptosis ,Biology ,Methylation ,Histones ,03 medical and health sciences ,chemistry.chemical_compound ,Histone H3 ,Mastocytosis, Systemic ,SETD2 ,Cell Line, Tumor ,medicine ,Humans ,Midostaurin ,Mast Cells ,Systemic mastocytosis ,Aged ,Bortezomib ,Lysine ,leukemia ,Hematology ,Histone-Lysine N-Methyltransferase ,Middle Aged ,medicine.disease ,Mast cell leukemia ,Prognosis ,Staurosporine ,3. Good health ,Settore MED/15 - MALATTIE DEL SANGUE ,030104 developmental biology ,Oncology ,chemistry ,Histone methyltransferase ,Mutation ,Cancer research ,Female ,Original Article ,K562 Cells ,Mastocytosis ,medicine.drug - Abstract
The molecular basis of advanced systemic mastocytosis (SM) is not fully understood and despite novel therapies the prognosis remains dismal. Exome sequencing of an index-patient with mast cell leukemia (MCL) uncovered biallelic loss-of-function mutations in the SETD2 histone methyltransferase gene. Copy-neutral loss-of-heterozygosity at 3p21.3 (where SETD2 maps) was subsequently found in SM patients and prompted us to undertake an in-depth analysis of SETD2 copy number, mutation status, transcript expression and methylation levels, as well as functional studies in the HMC-1 cell line and in a validation cohort of 57 additional cases with SM, including MCL, aggressive SM and indolent SM. Reduced or no SETD2 protein expression-and consequently, H3K36 trimethylation-was found in all cases and inversely correlated with disease aggressiveness. Proteasome inhibition rescued SETD2 expression and H3K36 trimethylation and resulted in marked accumulation of ubiquitinated SETD2 in SETD2-deficient patients but not in patients with near-normal SETD2 expression. Bortezomib and, to a lesser extent, AZD1775 alone or in combination with midostaurin induced apoptosis and reduced clonogenic growth of HMC-1 cells and of neoplastic mast cells from advanced SM patients. Our findings may have implications for prognostication of SM patients and for the development of improved treatment approaches in advanced SM.Leukemia advance online publication, 30 June 2017; doi:10.1038/leu.2017.183.
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- 2018
30. PS976 METABOLIC REPROGRAMMING OF ACUTE MYELOID LEUKEMIA STEM- PROGENITOR CELLS AT RELAPSE
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Daniel Remondini, Martina Ghetti, Cristina Papayannidis, Giorgia Simonetti, R. De Tommaso, Martina Pazzaglia, Maria Teresa Bochicchio, Maria Chiara Fontana, Jacopo Nanni, Samantha Bruno, Eugenia Franchini, Giovanni Martinelli, Anna Maria Ferrari, Roberta Napolitano, Antonella Padella, and Giovanni Marconi
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business.industry ,Metabolic reprogramming ,Cancer research ,Myeloid leukemia ,Medicine ,Hematology ,Progenitor cell ,business - Published
- 2019
31. S123 SETD2 LOSS OF FUNCTION IS A RECURRENT EVENT IN ADVANCED-PHASE CHRONIC MYELOID LEUKEMIA INDUCED BY POST-TRANSLATIONAL MECHANISMS THAT CAN BE THERAPEUTICALLY TARGETED
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Gabriele Gugliotta, Annalisa Imovilli, Patrizia Pregno, Marzia Salvucci, Francesco Albano, Claudia Baratè, Michele Baccarani, Alessandra Iurlo, B. Sinigallia, Mario Tiribelli, Elena Tenti, E. Abbruzzese, Elisa Dan, Fausto Castagnetti, S. De Santis, Monica Crugnola, Massimiliano Bonifacio, Manuela Mancini, Elisabetta Calistri, Luana Bavaro, Margherita Martelli, Michele Cavo, Sara Galimberti, Antonella Gozzini, Simona Soverini, Gianni Binotto, Giovanni Rosti, Cecilia Monaldi, Elena Trabacchi, Giovanni Martinelli, Maria Chiara Fontana, and Nicola Orofino
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Recurrent event ,Post translational ,business.industry ,SETD2 ,Advanced phase ,Cancer research ,Medicine ,Myeloid leukemia ,Hematology ,business ,Loss function - Published
- 2019
32. PS1437 SETD2 NON-GENOMIC LOSS OF FUNCTION IN ADVANCED SYSTEMIC MASTOCYTOSIS (SM): PATHOGENETIC AND THERAPEUTIC IMPLICATIONS
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Cecilia Monaldi, Giulia Paciello, Peter Valent, Michele Cavo, Michela Rondoni, Chiara Elena, Francesco Albano, Elisa Ficarra, S. De Santis, Fabio Ciceri, Cristina Papayannidis, Roberta Zanotti, Patrizia Tosi, Luana Bavaro, Maria Chiara Fontana, Margherita Martelli, Livio Pagano, Manuela Mancini, Massimo Delledonne, Giovanni Martinelli, and Simona Soverini
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SETD2 ,business.industry ,Immunology ,medicine ,Hematology ,Systemic mastocytosis ,medicine.disease ,business ,Loss function - Published
- 2019
33. Chromothripsis in acute myeloid leukemia: Biological features and impact on survival
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Michele Cavo, Zdenek Racil, Viviana Guadagnuolo, Antonella Padella, Maria Chiara Fontana, Michael Steurer, Michael Doubek, Emanuela Ottaviani, Nicoletta Testoni, Giovanni Marconi, Marco Manfrini, Torsten Haferlach, Carmen Baldazzi, Stefania Paolini, Simona Soverini, Andrea Ghelli Luserna di Rorà, Vincenza Solli, Robert Kralovics, Anna Maria Ferrari, Eugenio Fonzi, Cristina Papayannidis, Eugenia Franchini, Giovanni Martinelli, Lukáš Semerád, Ilaria Iacobucci, Jelena D. Milosevic Feenstra, Giorgia Simonetti, Fontana, Maria Chiara, Marconi, Giovanni, Feenstra, Jelena D. Milosevic, Fonzi, Eugenio, Papayannidis, Cristina, Ghelli Luserna Di Rorá, Andrea, Padella, Antonella, Solli, Vincenza, Franchini, Eugenia, Ottaviani, Emanuela, Ferrari, Anna, Baldazzi, Carmen, Testoni, Nicoletta, Iacobucci, Ilaria, Soverini, Simona, Haferlach, Torsten, Guadagnuolo, Viviana, Semerad, Luka, Doubek, Michael, Steurer, Michael, Racil, Zdenek, Paolini, Stefania, Manfrini, Marco, Cavo, Michele, Simonetti, Giorgia, Kralovics, Robert, and Martinelli, Giovanni
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0301 basic medicine ,Genome instability ,Oncology ,Adult ,Male ,medicine.medical_specialty ,NPM1 ,Cancer Research ,Adolescent ,Ring chromosome ,Polymorphism, Single Nucleotide ,Article ,03 medical and health sciences ,Young Adult ,Internal medicine ,Chromosome instability ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Biomarkers, Tumor ,Humans ,Genetic Predisposition to Disease ,Ring Chromosomes ,In Situ Hybridization, Fluorescence ,Aged ,Proportional Hazards Models ,Aged, 80 and over ,Chromothripsis ,Hematology ,business.industry ,Myeloid leukemia ,Adult Acute Myeloid Leukemia ,Middle Aged ,Prognosis ,3. Good health ,Chromosome Banding ,Leukemia, Myeloid, Acute ,030104 developmental biology ,Treatment Outcome ,Anesthesiology and Pain Medicine ,Mutation ,Female ,business ,Nucleophosmin - Abstract
Chromothripsis is a one-step genome-shattering catastrophe resulting from disruption of one or few chromosomes in multiple fragments and consequent random rejoining and repair. This study defines incidence of chromothripsis in 395 newly diagnosed adult acute myeloid leukemia (AML) patients from three institutions, its impact on survival and its genomic background. SNP 6.0 or CytoscanHD Array (Affymetrix??) were performed on all samples. We detected chromothripsis with a custom algorithm in 26/395 patients. Patients harboring chromothripsis had higher age (p???=???0.002), ELN high risk (HR) (p?????0.001), lower white blood cell (WBC) count (p???=???0.040), TP53 loss, and/or mutations (p?????0.001) while FLT3 (p???=???0.025), and NPM1 (p???=???0.032) mutations were mutually exclusive with chromothripsis. Chromothripsis-positive patients showed a worse overall survival (OS) (p?????0.001) compared with HR patients (p???=???0.011) and a poor prognosis in a COX-HR optimal regression model. Chromothripsis presented the hallmarks of chromosome instability [i.e., TP53 alteration, 5q deletion, higher mean of copy number alteration (CNA), complex karyotype, alterations in DNA repair, and cell cycle] and focal deletions on chromosomes 4, 7, 12, 16, and 17. CBA. FISH showed that chromothripsis is associated with marker, derivative, and ring chromosomes. In conclusion, chromothripsis frequently occurs in AML (6.6%) and influences patient prognosis and disease biology.
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- 2017
34. Chromothripsis in AML patients: A new mechanism of cancer initiation and progression
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Maria Chiara Fontana, Viviana Guadagnuolo, Cristina Papayannidis, Giovanni Marconi, Giorgia Simonetti, Antonella Padella, Marco Manfrini, Barbara Santacroce, Margherita Perricone, Silvia Lo Monaco, Emanuela Ottaviani, Simona Soverini, Michele Cavo, Giovanni Martinelli, and Maria Chiara Fontana, Viviana Guadagnuolo, Cristina Papayannidis, Giovanni Marconi, Giorgia Simonetti, Antonella Padella, Marco Manfrini, Barbara Santacroce, Margherita Perricone, Silvia Lo Monaco, Emanuela Ottaviani, Simona Soverini, Michele Cavo, Giovanni Martinelli
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Chromotripsis, Acute Myeloid Leukemia - Published
- 2016
35. New JAK2 heterozygous loss: A role in overall survival in acute myeloid leukemia patients
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Viviana Guadagnuolo, Maria Chiara Fontana, Cristina Papayannidis, Marco Manfrini, Antonella Padella, Giorgia Simonetti, Anna Ferrari, Giovanni Marconi, Andrea Ghelli Luserna di Rorà, Stefania Paolini, MariaChiara Abbenante, Sarah Parisi, Chiara Sartor, Emanuela Ottaviani, Giovanni Martinelli, and Viviana Guadagnuolo, Maria Chiara Fontana, Cristina Papayannidis, Marco Manfrini, Antonella Padella, Giorgia Simonetti, Anna Ferrari, Giovanni Marconi, Andrea Ghelli Luserna di Rorà, Stefania Paolini, MariaChiara Abbenante, Sarah Parisi, Chiara Sartor, Emanuela Ottaviani, Giovanni Martinelli
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JAK2, Acute Myeloid Leukemia - Published
- 2016
36. You have accessAlterations in Pathways Regulating Phosphatidil Inositol 3 Phosphate (PI3P) Produce Both Cell Proliferation and Therapy Resistance, and Define a Group of Patients with Poor Prognosis in Acute Myeloid Leukemia (AML)
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Giovanni Marconi, Cristina Papayannidis, Maria Chiara Fontana, Antonella Padella, Giorgia Simonetti, Marco Manfrini, Anna Ferrari, Eugenia Franchini, Stefania Paolini, Sarah Parisi, Chiara Sartor, Maria Chiara Abbenante, Silvia Lo Monaco, Francesca Volpato, Elena Tenti, Viviana Guadagnuolo, Margherita Perricone, Maria Teresa Bochicchio, Andrea Ghelli Luserna Di Rora, Carmen Baldazzi, Valentina Robustelli, Nicoletta Testoni, Simona Soverini, Emanuela Ottaviani, Giovanni Martinelli, and Giovanni Marconi, Cristina Papayannidis, Maria Chiara Fontana, Antonella Padella, Giorgia Simonetti, Marco Manfrini, Anna Ferrari, Eugenia Franchini, Stefania Paolini, Sarah Parisi, Chiara Sartor, Maria Chiara Abbenante, Silvia Lo Monaco, Francesca Volpato, Elena Tenti, Viviana Guadagnuolo, Margherita Perricone, Maria Teresa Bochicchio, Andrea Ghelli Luserna Di Rora, Carmen Baldazzi, Valentina Robustelli, Nicoletta Testoni, Simona Soverini, Emanuela Ottaviani, Giovanni Martinelli
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PI3P, Acute Myeloid leukemia - Published
- 2016
37. Microarray analysis to identifiy novel copy number alterations in acute myeloid leukemia
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Andrea Ghelli Luserna di Rorà, Luca Bertamini, Antonella Padella, Samantha Bruno, Valentina Robustelli, Carmen Baldazzi, Nicoletta Testoni, Eugenio Fonzi, Jacopo Nanni, Emanuela Ottaviani, Eugenia Franchini, Giovanni Martinelli, Maria Chiara Fontana, Giorgia Simonetti, Anna Maria Ferrari, Cristina Papayannidis, Maria Chiara Abbenante, Stefania Paolini, Giovanni Marconi, Fontana, MARIA CHIARA, Marconi, Giovanni, Papayannidis, Cristina, Fonzi, Eugenio, Ottaviani, Emanuela, Franchini, Eugenia, Ferrari, Anna, Simonetti, Giorgia, Padella, Antonella, Bruno, Samantha, Testoni, Nicoletta, Baldazzi, Carmen, GHELLI LUSERNA DI RORÀ, Andrea, Robustelli, Valentina, Abbenante, Mariachiara, Paolini, Stefania, Nanni, Jacopo, Bertamini, Luca, and Martinelli, Giovanni
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Cancer Research ,Tumor biology ,Oncology ,Microarray analysis techniques ,Myeloid leukemia ,Computational biology ,Biology ,Bioinformatics ,SNP array - Abstract
11622 Background: SNP microarray can detect Copy Number Alterations (CNAs) which could be predictive of response and can help define therapeutic strategies. Our aim is to improve conventional cytogenetic analysis and identify new genetic alterations relevant to leukemogenesis by a SNP array-based genotyping approach. Methods: We performed SNP 6.0/Cytoscan HD (Affymetrix) on 235 Acute Myeloid Leukemia (AML) patients at diagnosis. Seventy-eight/235 samples were also performed by Whole Exome Sequencing, WES (HiSeq,Illumina). SNP Array data were analyzed by Nexus Copy Number (BioDiscovery) and R Core Team. Results: We found several genes preferentially deleted, including MRPS5 (14.8%), PHF6 (9.3%), SCAPER (7.2%), CASK (5.9%), WNK (4.6%), STAG2 (4.2%), LRRK1 (3.4%), PALB2 (3.4%), genes preferentially amplified were RABL2B (16.1%), NF2 (10.2%), NBPF9 (7.6%), JAK2 (6.8%), RB1, NF1 and KMT2A (4.2%), PTEN (3.4%), TP73 and SMAD2 (2.5%). Single-copy losses and deletions were enriched (p < .001) for genes mapping in these pathways: aberrant PD-1 signaling, loss of function of SMAD4 in cancer and SMAD4 MH2 Domain mutants in cancer. The pathways significantly (p < .001) deregulated in our cohort with single copy gain and homozygous amplification were: regulation of transcription and nucleic acid, negative regulation of metabolic processes, constitutive signaling by aberrant PI3K in cancer and PI3K/AKT network. In order to define driver alterations, we correlate deletions and losses with mutational data. We found losses are also targeted by mutations ( BRCA2, LRRK1), while deleted genes, as CASK, CDK6 and MAPT, were involved in pathways affected by genomic mutations ( CASK deletion and MPP6 mutation, CDK6 deletion and PPM1D mutation, MAPT deletion and SPAG5mutation). Conclusions: We have identified new CNAs and pathways involving novel potential leukemia-related genes. The comparison between SNP and WES data could provide important findings on prognosis of AML patients. Minimal deleted regions of genes in deregulated pathways deserve further investigation in order to identify new genes which could be relevant AML biomarkers. Ackn: ELN, AIL, AIRC,prog. Regione-Università 2010-12 (L. Bolondi),FP7 NGS-PTL project,HARMONY.
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- 2017
38. Prognostic significance of alterations of pathways regulating autophagy in acute myeloid leukemia
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Giovanni Marconi, Maria Chiara Fontana, Cristina Papayannidis, Antonella Padella, Silvia Lo Monaco, Maria Chiara Abbenante, Chiara Sartor, Luca Bertamini, Jacopo Nanni, Andrea Ghelli Luserna di Rorà, Valentina Robustelli, Elena Tenti, Eugenio Fonzi, Giorgia Simonetti, Emanuela Ottaviani, Carmen Baldazzi, Nicoletta Testoni, Giovanni Martinelli, Marconi, Giovanni, Fontana, MARIA CHIARA, Papayannidis, Cristina, Padella, Antonella, LO MONACO, Silvia, Abbenante, Mariachiara, Sartor, Chiara, Bertamini, Luca, Nanni, Jacopo, GHELLI LUSERNA DI RORÀ, Andrea, Robustelli, Valentina, Tenti, Elena, Fonzi, Eugenio, Simonetti, Giorgia, Ottaviani, Emanuela, Baldazzi, Carmen, Testoni, Nicoletta, and Martinelli, Giovanni
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0301 basic medicine ,Cancer Research ,Leukemia ,business.industry ,Mechanism (biology) ,Autophagy ,Cancer ,Myeloid leukemia ,medicine.disease ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Oncology ,Apoptosis ,030220 oncology & carcinogenesis ,Cancer research ,Medicine ,Treatment resistance ,business - Abstract
7038 Background: Nowadays, science is debating if autophagy in cancer can lead to therapy resistance or it can favor apoptosis. Autophagy pathways are involved pro-apoptotic mechanism, or they can improve stresses survival eliminating damaged mitochondria and proteins. Levels and activity of pro-apoptotic and anti-apoptotic proteins (eg. bcl-2 and p53), high levels of cAMP, and a pink/park complex could play as fulcrum on this lever. Our study aims to define the role of autophagy in AML. Methods: We analyzed 148 consecutive non M3 AML with Affymetrix SNP array. We screened all patients for TP53, FLT3, NMP1 mutations. Patients was treated with intensive induction chemotherapy regimens. Survival data were collected prospectively, with a median follow-up of 18 months. Results: Autophagy alteration (gene group 1: ULK1 CHR11; ULK1 CHR17; BECN1; ATG14; AMBRA1; UVRAG; ATG9A; ATG9B; PIK3C3; PIK3R4) was related to lower Complete Remission rate (CR%) after induction in univariate (p < .001) and multivariable regression model with age, karyotype, secondary AML, TP53 mutation (p = .014); autophagy alteration shown to confer worst Overal Survival (OS) (p < .001) and was significantly associated with complex karyotype and TP53 mutation (p < .001). We detected significant differences in term of survival independently both in gain and loss in group 1 genes (p < .001). Alterations in genes in cAMP pathway (group 2: SESN1; PRKAA1 CHR 3; PRKAB1: PRKAA1 CHR 1: PRKAG1 CHR11; PRKAG1 CHR 7; PRKAG3; PRKAB1) and in genes that could be related to a switch from a physiological role of autophagy to a resiliency mechanism (group 3: CCND1; BCL2; PINK1; PARK2; TP53; MDM1; MDM4) showed to confer worst OS (p < .001 in both groups); Alteration in group 2 and group 3 were related to lower CR% after induction (p < .001 in both groups). Whole Exome Sequencing on 56 patients in our set did not found any significant mutation in genes we analyzed with the exception of TP53. Conclusions: Alterations in autophagy regulator genes are associated with poor prognosis and therapy resistance. A loss in autophagy could block apoptosis, a gain could confer cell resiliency. Acknowledgements: ELN, AIL, AIRC, Progetto Regione-Università 2010-12,FP7 NGS-PTL, HARMONY
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- 2017
39. Deficient necroptosis pathway as a negative prognostic factor in acute myeloid leukemia
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Antonella Padella, Sarah Parisi, Carmen Baldazzi, Eugenio Fonzi, Nicoletta Testoni, Giovanni Marconi, Maria Chiara Fontana, Valentina Robustelli, Cristina Papayannidis, Luca Bertamini, Anna Maria Ferrari, Andrea Ghelli Luserna di Rorà, Silvia Lo Monaco, Jacopo Nanni, Eugenia Franchini, Giovanni Martinelli, Emanuela Ottaviani, Stefania Paolini, Maria Chiara Abbenante, Giorgia Simonetti, LO MONACO, Silvia, Marconi, Giovanni, Fontana, MARIA CHIARA, Papayannidis, Cristina, Fonzi, Eugenio, Baldazzi, Carmen, Testoni, Nicoletta, Ottaviani, Emanuela, Franchini, Eugenia, Ferrari, Anna, Simonetti, Giorgia, Padella, Antonella, GHELLI LUSERNA DI RORÀ, Andrea, Robustelli, Valentina, Paolini, Stefania, Abbenante, Mariachiara, Parisi, Sarah, Nanni, Jacopo, Bertamini, Luca, and Martinelli, Giovanni
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Cancer Research ,Prognostic factor ,Tumor biology ,business.industry ,Necroptosis ,Myeloid leukemia ,Necrotic cell ,Oncology ,Interferon ,Transcription (biology) ,medicine ,Cancer research ,Receptor ,business ,Gene ,medicine.drug - Abstract
11611 Background: Necroptosis is a type of necrotic cell death involving several genes transcription and activation of molecular mechanisms as death receptors, interferon, toll-like receptors, intracellular RNA and DNA sensors.The process is leading by the family of receptor-interacting protein kinase ( RIPK3, RIPK2, RIPK1) and the MLKL substrate. Losses of RIPK3 or MLKL, as well as deficiency in apoptosis, could allow tumor cells to escape the immunomediated cells death (ICD). Methods: We performed SNP Arrays (Cytoscan HD and SNP 6.0, Affymetrix) on a cohort of 300 non-M3 AML patients at diagnosis and we analyzed the Overall Survival (OS) of our patients with deficiency on necroptosis pathways. Survival was analyzed with Kaplan-Mayer method and Log-Rank test. We further analyze the relevance of different prognostic factors by the use of COX-Hazard Ratio statistical analysis. Results: We find that 18 patients presented a loss of RIPK1 or MLKL (nobody presented losses in RIPK3/RIPK2) and 13/18 patients were older than 65 years old. The Overall Survival (OS) of patients with alterations in these genes is significantly lower than control group, with a median OS of 3 vs 6 month respectively (p
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- 2017
40. Abstract 2964: Pharmacological inhibition of WIP1 by GSK2830371 sensitizes AML cells to MDM2 inhibitor Nutlin-3a
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Andrea Ghelli Luserna di Rorà, Martina Pazzaglia, Antonella Padella, Giorgia Simonetti, Michele Cavo, Simona Soverini, Giovanni Martinelli, Jacopo Nanni, Giovanni Marconi, Enrica Imbrogno, Matteo Bocconcelli, Ilaria Iacobucci, Maria Chiara Fontana, Cristina Papayannidis, Anna Maria Ferrari, and Maria Teresa Bochicchio
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Cancer Research ,biology ,medicine.diagnostic_test ,DNA repair ,Chemistry ,In vitro ,Oncology ,Downregulation and upregulation ,Western blot ,Apoptosis ,Cell culture ,Annexin ,Cancer research ,biology.protein ,medicine ,Mdm2 - Abstract
Introduction: PPM1D (wild-type p53-inducible protein phosphatase WIP1) is a member of the PP2C family serine/threonine phosphatase involved in negative regulation of cell stress response pathways, leading to suppression of p53 after stress. To restore p53 function through MDM2 inhibition (Nutlin-3a) is a promising approach in AML. Promising results of the combination of MDM2 inhibitors and WIP1 inhibitor (WIP1i), obtained in many preclinical studies of solid tumors, paved the way for their application in AML. We investigated whether the inhibition of WIP1 (GSK2830371) could sensitize AML cell lines and primary cells to Nutlin-3a (Nut-3a) in order to obtain a novel therapeutic strategy for AML patients, based on restoration of p53 activity. Methods: In vitro viability (by WST-1 reagent) and Annexin-V/PI apoptosis assay were performed on a panel of TP53-wt AML cells (MOLM-13, MV-4-11 and OCI-AML3), on TP53-mutated NOMO-1 AML cells and on primary AML samples. Gene expression profile (GEP) and Western Blot (WB) analyses were performed on MV-4-11 and NOMO-1 after 16h. Results: Combined inhibition of increasing dosage of Nut-3a (0.5 to 5 uM) and WIP1i (5 to 20 uM) synergistically reduces TP53-wt AML cells viability, while NOMO-1 resulted to be insensitive to the combination. The combination index analyses showed a synergistically effect of the combination of both compounds on TP53-wt AML cells. Annexin V/PI staining showed that WIP1i sensitizes TP53-wt AML cell lines and primary samples to Nut-3a-induced apoptosis, when compared with single treatment. No effect was seen in NOMO-1. GEP demonstrated that MV-4-11 cells exhibits a major response to drug combination with an upregulation of cell cycle control genes, a downregulation of DNA repair machinery genes, upregulation of MDM2 and of TP53-downstream genes (eg.CDKN1A), confirming the activation of p53 pathway. NOMO-1 cells showed upregulation of resiliency-mechanism and confirmed insensitivity to both treatment. WB analysis confirmed GEP data showing an increased expression of p53 and p21 in wt-TP53 cell line after 16h of combined-treatment. Conclusions: We identified a novel synergistic drug combination between Nut-3a and WIP1i that induces apoptosis in AML cell lines and primary samples. GEP and WB of TP53-wt MV-4-11 and TP53-mutated NOMO-1 cells showed mechanisms underlying drug sensitivity and resistance giving novel insights on potential markers of response and novel drugable targets. In vivo studies are needed to confirm these preclinical data. Supported by: AIRC, FP7-NGS-PTL, Fondazione del Monte, HARMONY. Citation Format: Maria Chiara Fontana, Jacopo Nanni, Giovanni Marconi, Martina Pazzaglia, Matteo Bocconcelli, Antonella Padella, Simona Soverini, Ilaria Iacobucci, Cristina Papayannidis, Anna Ferrari, Maria Teresa Bochicchio, Enrica Imbrogno, Michele Cavo, Andrea Ghelli Luserna di Rora, Giorgia Simonetti, Giovanni Martinelli. Pharmacological inhibition of WIP1 by GSK2830371 sensitizes AML cells to MDM2 inhibitor Nutlin-3a [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2964.
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- 2019
41. Abstract 3100: Blinatumomab is safe and effective in relapsed and MRD positive B-ALL CD19+ patients: The bologna compassionate program experience
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Giovanni Martinelli, Stefania Paolini, Claudio Cerchione, Alessandra Santoro, Valentina Robustelli, Simona Soverini, Caterina De Benedittis, Enrica Imbrogno, Andrea Ghelli Luserna Di Rora, Sarah Parisi, Chiara Sartor, Giovanni Marconi, Silvia Lo Monaco, Maria Chiara Abbenante, Maria Chiara Fontana, Antonella Padella, Anna Ferrari, Giorgia Simonetti, Elena Tenti, Federica Frabetti, Francesca Volpato, Samantha Bruno, Fabiana Mammoli, Maria Teresa Bocchicchio, Carmen Baldazzi, and Cristina Papayannidis
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Cancer Research ,Oncology - Abstract
Adult B-ALL patients still have a dismal prognosis, due to a high incidence of relapse even after allogenic SCT. Safety and efficacy of Blinatumomab, an anti CD3-CD19 Bite antibody, has been demonstrated both in MRD positive patients and in relapsed/refractory (R/R) setting. To evaluate safety profile and efficacy of Blinatumomab, obtained through a compassionate use, in a cohort of 18 adult patients affected by MRD+ or R/R B-ALL treated at Bologna University. From March 2015 to December 2017, 18 patients received Blinatumomab at the standard dosage (9 mcg/d x 7 days, 28 mcg/d x 21 days) in 28-days courses. All the patients were hospitalized to receive the first course of therapy. The following courses, based on the good safety profile of the compound, were administered in outpatient setting. 18 patients (M/F = 10/8; median age 43, range 18-73) have been treated. Philadelphia (Ph) chromosome was detected in 8/18 patients. 10 patients were MRD+ (5 Ph pos and 5 Ph neg); E2A-PBX1 and MLL-AF4 rearrangements were found in two patients. 8 patients had a R/R disease (3 Ph pos and 5 Ph neg). Median WBC count before starting therapy with Blinatumomab was 5400/mmc (range 500-76500). All the patients had previously received many lines of therapy (median 4, range 1-7). In 4 cases an alloTMO was already performed, and two patients had received two transplants. 12/18 patients were referred to us by other Italian Institutions. All the patients received at least one course of Blinatumomab. In one case three courses were administered; an elderly patient is actually receiving the fifth course. Globally, 32 courses of therapy have been administered (median 2, range 1-5). Bone marrow evaluations, including cytogenetics, molecular biology and immunophenotyping analysis were performed at the beginning of every course of therapy in order to assess patients' disease status. MRD evaluation was assessed through BCR-ABL fusion transcript quantitative analysis in Ph pos ALL patients and Ig rearrangment in Ph negative patients. Monitoring of adverse events was periodically performed. 16/18 patients are evaluable for response, at least to one cycle (one patient died during the first course, one patient is still receiving the first course). 9/16 (56%) patients obtained a CR (7/9 MRD+ and 2/7 R/R). In 7/9 (78%) responders patients a molecular CR was reached, (in 6 patients after the first course, in one case after the second one). 5 responders proceeded to alloBMT and are actually alive in CR (median follow-up after transplant 240 days). In terms of toxicity, one patient developed a grade IV neurological event (mental confusion, tremor), which completely resolved after a transient drug withdrawal. Our results confirm the high rate of response and to Blinatumomab in a poor patients' population, and the good management profile of the compound. Note: This abstract was not presented at the meeting. Citation Format: Giovanni Martinelli, Stefania Paolini, Claudio Cerchione, Alessandra Santoro, Valentina Robustelli, Simona Soverini, Caterina De Benedittis, Enrica Imbrogno, Andrea Ghelli Luserna Di Rora, Sarah Parisi, Chiara Sartor, Giovanni Marconi, Silvia Lo Monaco, Maria Chiara Abbenante, Maria Chiara Fontana, Antonella Padella, Anna Ferrari, Giorgia Simonetti, Elena Tenti, Federica Frabetti, Francesca Volpato, Samantha Bruno, Fabiana Mammoli, Maria Teresa Bocchicchio, Carmen Baldazzi, Cristina Papayannidis. Blinatumomab is safe and effective in relapsed and MRD positive B-ALL CD19+ patients: The bologna compassionate program experience [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3100.
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- 2019
42. Abstract 5279: Metabolic profiling defines a new characterization of acute myeloid leukemia and identifies NPM1-mutated cases as a distinct subgroup
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Samantha Bruno, Anna Maria Ferrari, Rossella De Tommaso, Italo Faria do Valle, Jacopo Nanni, Margherita Perricone, Maria Chiara Fontana, Martina Pazzaglia, Antonella Padella, Maria Teresa Bochicchio, Emanuela Ottaviani, Cristina Papayannidis, Claudio Cerchione, Eugenia Franchini, Giovanni Martinelli, Enrica Imbrogno, Giorgia Simonetti, Giovanni Marconi, Daniel Remondini, and Eugenio Fonzi
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0301 basic medicine ,Cancer Research ,NPM1 ,IDH1 ,CD33 ,CD34 ,Myeloid leukemia ,Biology ,medicine.disease_cause ,Molecular biology ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Oncology ,030220 oncology & carcinogenesis ,medicine ,KRAS ,Gene ,Exome sequencing - Abstract
Genomic and functional alterations of enzymatic activity drive cancer metabolic reprogramming along with mutations of tumor suppressors and oncogenes. IDH1/2 lesions represent a paradigmatic example in acute myeloid leukemia (AML). The study aimed to stratify AML patients based on their metabolic landscape and to map potential connections between their metabolic and genomic profile. The genomic landscape of 166 AML patients was obtained by whole exome sequencing. Variants were called by MuTect and Varscan2. Six additional cases were analyzed by targeted sequencing (SOPHiA GENETICS). Metabolites were quantified by mass spectrometry of bone marrow cells (35 CD34+ and 15 CD33+ AML from the above-mentioned cohort) and compared with CD34+ cord blood and CD33+ healthy blood cells (n=21 each, Metabolon) by Welch's t-test. In AML, 17% of somatic variants targeted metabolism-related genes: 38% were enzymes (according to Recon2), including 3% of electron transport chain genes encoded by mitochondrial DNA (COX1-3, ND1-6), 3% were AML-related genes with a known involvement in cell metabolism (e.g. IDH1/2, MYC, KRAS) and 59% were metabolic regulators, defined by gene ontology annotation. Ninety-one% of patients carried at least one mutation in a metabolism-related gene, with 43% of variants rated as damaging. The most represented pathways were lipid, carbohydrate, nucleotide (AK9, H6PD), amino acids (IDO2) and glucose metabolism (PKM, HK3). Principal component analysis of metabolic data showed a distinct profile between AML and healthy cells, with a predictive accuracy of 86% and 95% for CD34+ and CD33+ cells, respectively. Conversely, few differences were observed between CD34+ and CD33+ AML. Unsupervised hierarchical clustering clearly defined 3 AML clusters (C1-3). Moreover, 3 subgroups could be identified in C3 without ambiguous assignments. C1 was enriched for NPM1-mutated (mut) cases (83%, 33%, 27% in C1, C2 and C3, respectively, p=0.03). NPM1-mutated AML were distinguished by a 12-metabolites signature. They showed increased levels of spermidine, cytidine 2′ or 3′-monophosphate (P), thymidine 3′-monoP, uridine-2',3'-cyclic monoP and decrease of inosine 5'-monoP, suggesting altered polyamine, pyrimidine and purine metabolism. Moreover, NPM1-mut AML had reduced levels of intermediates involved in acyl carnitine, lysophospholipid, phosphatidylethanolamine and sphingolipid metabolism. Overall, mutations of metabolism-related genes are common in AML. We defined a metabolic-based classification of AML and identified a new metabolic signature based on 12 metabolites that distinguish NPM1-mut AML from wild-type cases and healthy CD34+/CD33+ cells. Major alterations in the nucleotide and polyamine pathways suggest novel potential therapeutic approaches. Supported by: EHA research fellowship award, AIRC, FP7-NGS-PTL, Fondazione del Monte. Citation Format: Giorgia Simonetti, Antonella Padella, Eugenio Fonzi, Martina Pazzaglia, Margherita Perricone, Maria Chiara Fontana, Samantha Bruno, Maria Teresa Bochicchio, Eugenia Franchini, Jacopo Nanni, Giovanni Marconi, Italo F. do Valle, Rossella De Tommaso, Anna Ferrari, Enrica Imbrogno, Claudio Cerchione, Cristina Papayannidis, Emanuela Ottaviani, Daniel Remondini, Giovanni Martinelli. Metabolic profiling defines a new characterization of acute myeloid leukemia and identifies NPM1-mutated cases as a distinct subgroup [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5279.
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- 2019
43. Acute Myeloid Leukemia Mutations: Therapeutic Implications
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Giovanni Marconi, Sarah Parisi, Chiara Sartor, Gianluca Cristiano, Antonio Curti, Maria Chiara Fontana, Stefania Paolini, Cristina Papayannidis, and Jacopo Nanni
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0301 basic medicine ,Review ,Disease ,acute myeloid leukemia ,Bioinformatics ,Catalysis ,lcsh:Chemistry ,resistance ,Inorganic Chemistry ,Pathogenesis ,03 medical and health sciences ,Therapeutic approach ,0302 clinical medicine ,Gene Frequency ,Antineoplastic Combined Chemotherapy Protocols ,Biomarkers, Tumor ,Humans ,Medicine ,Genetic Predisposition to Disease ,Molecular Targeted Therapy ,Physical and Theoretical Chemistry ,FLT3 ,lcsh:QH301-705.5 ,Protein Kinase Inhibitors ,Molecular Biology ,Alleles ,Spectroscopy ,Clinical Trials as Topic ,business.industry ,Organic Chemistry ,Myeloid leukemia ,General Medicine ,mutations ,Computer Science Applications ,Transplantation ,Leukemia, Myeloid, Acute ,Treatment Outcome ,030104 developmental biology ,Isocitrate dehydrogenase ,lcsh:Biology (General) ,lcsh:QD1-999 ,030220 oncology & carcinogenesis ,Mutation ,Hematological neoplasm ,IDH1-2 ,Stem cell ,business - Abstract
Acute Myeloid Leukemia (AML) is an extremely heterogeneous group of hematological neoplasms, for which allogeneic stem cell transplantation (HSCT) still represents the only potentially curative option in the majority of cases. However, elderly age and clinically severe comorbidities may often exclude a wide amount of patients from this therapeutic approach, underlying the urgent need for alternative strategies. Thanks to the introduction of advanced high-throughput techniques, light is being shed on the pathogenesis of AML, identifying molecular recurrent mutations as responsible for the onset, as well as progression, of disease. As a consequence, and in parallel, many new compounds, including targeted therapies (FMS-like tyrosine kinase 3 (FLT3) and Isocitrate dehydrogenase 1-2 (IDH1-2) inhibitors), have found a wide room of application in this setting, and are now available in daily practice, or in late phases of clinical development. Moreover, several further innovative molecules are currently under investigation, and promising results for many of them have already been reported. In this review, we will present an update on the most relevant molecular alterations of AML, focusing on the most frequent genomic mutations of the disease, for which compounds have been approved or are still currently under investigation.
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- 2019
44. PB1726 DOUBLE FLUDARABINE-BASED INDUCTION AND INFECTIVE RISK: THE BOLOGNA EXPERIENCE
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Maria Chiara Fontana, Jacopo Nanni, Antonio Curti, Sarah Parisi, M. Ottaviani, S. De Polo, L. Bandini, Chiara Sartor, M. Stanzani, Annalisa Talami, Stefania Paolini, Giovanni Martinelli, Matteo Olivi, Mariachiara Abbenante, R.E. Lewis, Luca Bertamini, Michele Cavo, Simone Ragaini, Cristina Papayannidis, Giovanni Marconi, and Gianluca Cristiano
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Oncology ,medicine.medical_specialty ,business.industry ,Internal medicine ,medicine ,Hematology ,business ,Fludarabine ,medicine.drug - Published
- 2019
45. Aurora Kinase a/MDM2-Mediated SETD2 Loss of Function in Chronic Myeloid Leukemia Patients in Blast Crisis Induces Genetic Instability and Can be Therapeutically Targeted
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Mario Tiribelli, Massimiliano Bonifacio, Barbara Sinigaglia, Annalisa Imovilli, Simona Soverini, Giovanni Martinelli, Antonella Gozzini, Alessandra Iurlo, Gabriele Gugliotta, Maria Chiara Fontana, Elisa Dan, Michele Baccarani, Elisabetta Calistri, Elisabetta Abruzzese, Luana Bavaro, Margherita Martelli, Monica Crugnola, Gianantonio Rosti, Manuela Mancini, Marzia Salvucci, Sara Galimberti, Claudia Baratè, Fausto Castagnetti, Elena Trabacchi, Patrizia Pregno, Cecilia Monaldi, Michele Cavo, Francesco Albano, Sara De Santis, Nicola Orofino, Elena Tenti, and Gianni Binotto
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Blast Crisis ,biology ,business.industry ,Immunology ,Myeloid leukemia ,Cell Biology ,Hematology ,Biochemistry ,SETD2 ,biology.protein ,Cancer research ,Medicine ,Mdm2 ,Aurora Kinase A ,business ,Loss function - Abstract
The SETD2 protein is a histone methyltransferase that specifically catalyzes the trimethylation of Lysine 36 on histone H3 (H3K36me3). SETD2/H3K36me3 are implicated in transcript elongation and splicing, DNA repair, chromosome segregation. SETD2 gene deletions and/or mutations (mostly frameshift or nonsense) have been reported in solid tumors (clear cell renal cell carcinoma, bladder cancer, lung cancer, melanoma, endometrial cancer) and in acute leukemias. Using a Western Blotting (WB) approach to screen for SETD2 protein expression and for H3K36me3 levels in a relatively large cohort of 80 advanced-phase chronic myeloid leukemia (CML) patients (pts), we could detect reduced or null SETD2 and H3K36me3 in 86% of pts as compared to a pool of healthy donors and to chronic phase (CP) pts at diagnosis who achieved optimal responses to TKI, but neither mutations/deletions nor transcriptional down-regulation were the underlying causes. Inhibition of proteasome-mediated degradation in primary cells from pts with undetectable SETD2 restored H3K36me3 and led to accumulation of hyper-ubiquitinated SETD2, suggesting that a functional protein is produced but rapidly degraded. Moreover, proteasome inhibition was found to induce apoptosis and to reduce clonogenic growth. In K562 cells (SETD2/H3K36me3low), co-immunoprecipitation (co-IP) performed before and after proteasome inhibition showed accumulation of the hyper-ubiquitinated form of SETD2 bound to MDM2. MDM2 inhibition by SP-141 resulted in cytostatic effects and restored SETD2 expression and activity. Superimposable results were achieved by siRNA-mediated silencing of MDM2, suggesting that MDM2 is implicated in SETD2 reduced stability. Co-IP also showed that SETD2 interacts with Aurora Kinase A a Ser-Thr kinase frequently overexpressed in CML. We found that Aurora Kinase A phosphorylates SETD2, and both pharmacological inhibition by Danusertib and siRNA-mediated silencing rescued SETD2 expression and activity. Next, to investigate whether SETD2/H3K36me3 loss may contribute to genetic instability, LAMA 84 (SETD2/H3K36Me3high) and K562 (SETD2/H3K36me3low) cells were studied by WB and immunofluorescence (IF) to assess phosphorylated histone 2A.X (γH2AX) and Rad51 foci in steady state conditions and after sub-lethal DNA damage by UV exposure. The same studies were performed after SETD2 silencing for 3 months. Cells with low or silenced SETD2 had significantly higher levels of γH2AX and were unable to induce homologous recombination (HR) repair after DNA damage. Clonogenic assays performed in LAMA 84 cells before and after SETD2 silencing, in K562 (SETD2/H3K36me3low) and in imatinib-resistant (IM-R) K562 cells which have lost SETD2 expression and activity, suggested that reduction of clonogenic growth after proteasomal or MDM2 inhibition is strictly dependent on SETD2 expression and functional status (Figure 1A). First and second generation proteasome inhibitors (bortezomib, carfilzomib and ixazomib) inhibited the clonogenic potential of the mononuclear cell fraction from both CP (n=2) and blast crisis (BC) (n=4) CML pts at subnanomolar concentrations, with the extent of anti-tumor activity clearly anti-correlated with SETD2 expression and H3K36me3 levels: pts with lower SETD2 expression showed lower LD50 when compared with pts with higher SETD2 expression and H3K36me3 levels (Figure 1B). Similarly, clonogenic assays performed by administrating increasing doses of SP-141 (from 0.25 to 1.25 µM) suggested that MDM2 specific inhibition had more significant effects in BC-CML pts showing low SETD2 levels and activity as compared to BC-CML pts showing intermediate SETD2 levels and activity and to CP CML pts. In conclusion, phosphorylation by Aurora Kinase A and ubiquitination by MDM2 contribute to SETD2 non-genomic loss of function in advanced-phase CML. Loss of SETD2/H3K36me3 is associated with increased DNA damage and impaired HR repair. Restoring physiological H3K36me3 levels may help improve the outcome of this critical subset of pts. Acknowledgments: study supported by AIRC (project code 16996) and AIL (Associazione Italiana contro le Leucemia, Linfomi e Mieloma). Figure 1. Figure 1. Disclosures Castagnetti: Incyte: Consultancy, Honoraria; Pfizer: Consultancy, Honoraria; Bristol Meyers Squibb: Consultancy, Honoraria; Novartis: Consultancy, Honoraria. Gugliotta:Novartis: Honoraria; Pfizer: Honoraria; Bristol-Myers Squibb: Honoraria; Incyte: Honoraria. Abruzzese:Pfizer: Consultancy; Novartis: Consultancy; BMS: Consultancy; Ariad: Consultancy. Bonifacio:Incyte: Consultancy; Pfizer: Consultancy; Amgen: Consultancy; Novartis: Research Funding; Bristol Myers Squibb: Consultancy. Martinelli:Ariad/incyte: Consultancy; Pfizer: Consultancy; Celgene: Consultancy; Amgen: Consultancy; Janssen: Consultancy; Roche: Consultancy. Cavo:Adaptive Biotechnologies: Honoraria, Membership on an entity's Board of Directors or advisory committees; GlaxoSmithKline: Honoraria, Membership on an entity's Board of Directors or advisory committees; Bristol-Myers Squibb: Honoraria, Membership on an entity's Board of Directors or advisory committees; Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees; AbbVie: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees. Soverini:Bristol Myers Squibb: Consultancy; Incyte Biosciences: Consultancy; Novartis: Consultancy.
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- 2018
46. Higher Expression of PALB2 Predict Poor Prognosis in AML Patients and Identifies Potential Targets of Synthetic Lethal Therapies
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Giorgia Simonetti, Antonella Padella, Emanuela Ottaviani, Andrea Ghelli Luserna di Rorà, Simona Soverini, Giovanni Marconi, Cristina Papayannidis, Michele Cavo, Maria Chiara Fontana, and Giovanni Martinelli
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Poor prognosis ,Expression (architecture) ,business.industry ,PALB2 ,Immunology ,Cancer research ,Medicine ,Cell Biology ,Hematology ,business ,Biochemistry - Abstract
Background - Partner and localizer of BRCA2 (PALB2) plays a key role in the DNA damage repair (DDR). Genomic alterations of DDR genes rarely occur in AML, while their deregulation at transcriptional level is a known mechanism exploited by leukemic cells in order to sustain the high genetic instability and to continue proliferation. Aim - We aimed to characterize the role of PALB2 in AML by investigating its expression levels and its prognostic value, in order to evaluate its potential as target of therapies based on a synthetic lethality approaches. Methods - Gene expression profiling (GEP, Affymetrix) was performed on bone marrow cells of 7 healthy donors (HD) and 60 AML patients with more than 80% blast cells. K-means clustering of patients according to the expression of PALB2 was performed and differences in survival were assessed d by Kaplan-Meier survival analysis. Results - Our cohort was characterized by a median age at diagnosis of 60 years-old. Twelve out of 43 patients harbored a mutation in FLT3 (27.9%, 17 patients not tested); 8 patients were NPM1 mutated (27.6%, 31 patients not tested); 2 patients were TP53 mutated (3%, all patients tested). According to ELN2017 guidelines, 17 cases were high-risk AML, 38 cases were intermediate, 3 cases had low-risk classification and 2 cases were not classified due to the lack of prognostic markers. We detected variable levels of PALB2 mRNA (range 52.90-244.37) in AML patients and its median expression was higher compared to HD (129.26 vs 67.85, respectively; p=.019). We clustered our patients according to PALB2 expression and we defined 2 groups of patients: cluster H and L with higher and lower expression levels of PALB2, respectively (cluster centers 158.86 and 105.41, respectively; figure A). Notably, HD revealed PALB2 expression values comparable with the cluster L (range 52.90-99.60). No differences were detected in term of incidence mutations in FLT3 and NPM1, white blood cell count at diagnosis, age at diagnosis and prevalence of karyotype alterations. Patients were treated with best supportive therapy (n=11/58, therapy data missing for 2 patients), hypomethylating agents (n=2/58) and intensive chemotherapy ( n=45/58). Within patients treated with intensive chemotherapy, we compared complete remission rate after induction and we found no differences between H and L. However, patients with higher expression of PALB2 had worst overall survival than patients in cluster L (median survival group H=397 days; CI 95%=288.9-505.0, L=not reached; p=.045; figure B). Notably, we confirmed worst prognosis in patients in cluster H when considering 33 out of 45 patients with intermediate/low risk karyotype (i.e. normal karyotype, t(8;21) or less than 3 aberrations according to ELN2017; p=.026). Conclusion - We identified a subgroup of AML patients with higher expression of PALB2, which predicted poorer prognosis in patients treated with curative intent and it associated with poorer prognosis in patients with low/intermediate risk. While patients carrying mutations in PALB2 (and BRCA1/2) are candidate for PARP inhibitors (PARPi) therapies in breast cancers, few clinical trials with PARPi are available in AML and the frequency of mutations is very low. Our data opens a new scenario in which PALB2 may be a target of therapies in AML based on synthetic lethal approaches targeting the DDR pathway. However, a better understanding of the biological role of PALB2 in AML and its interaction with other alterations is needed. Supported by Fondazione Del Monte Figure. Figure. Disclosures Cavo: Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; AbbVie: Honoraria, Membership on an entity's Board of Directors or advisory committees; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees; Adaptive Biotechnologies: Honoraria, Membership on an entity's Board of Directors or advisory committees; Bristol-Myers Squibb: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; GlaxoSmithKline: Honoraria, Membership on an entity's Board of Directors or advisory committees. Soverini:Novartis: Consultancy; Bristol Myers Squibb: Consultancy; Incyte Biosciences: Consultancy. Martinelli:Roche: Consultancy; Celgene: Consultancy, Speakers Bureau; Janssen: Consultancy; Ariad/Incyte: Consultancy; Amgen: Consultancy; Pfizer: Consultancy, Speakers Bureau; Abbvie: Consultancy; Jazz Pharmaceuticals: Consultancy; Novartis: Speakers Bureau.
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- 2018
47. Biology of Acute Myeloid Leukemia (AML) with Monosomy of Chromosome 7 or Loss of 7q. a Study on 487 Patients Analyzed By Gene Expression Profile (GEP), Single Nucleotide Polymorphism (SNP) Arrays and Metabolomics
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Sarah Parisi, Jacopo Nanni, Chiara Sartor, Maria Chiara Abbenante, Simona Soverini, Annalisa Talami, Luca Bertamini, Nicoletta Testoni, Torsten Haferlach, Samantha Bruno, Maria Teresa Bochicchio, Antonio Curti, Simone Ragaini, Stefano De Polo, Anna Maria Ferrari, Matteo Olivi, Eugenio Fonzi, Giovanni Marconi, Michele Cavo, Giovanni Martinelli, Cristina Papayannidis, Maria Chiara Fontana, Robert Kralovics, Emanuela Ottaviani, Martina Pazzaglia, Stefania Paolini, Carmen Baldazzi, Jelena D. Milosevic Feenstra, and Giorgia Simonetti
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Chromosome 7 (human) ,Monosomy ,Immunology ,Myeloid leukemia ,Single-nucleotide polymorphism ,Cell Biology ,Hematology ,Biology ,medicine.disease ,Biochemistry ,Gene expression profiling ,Leukemia ,medicine ,Chromosome abnormality ,Cancer research ,Cell aging - Abstract
Introduction Monosomy 7 (-7) and interstitial deletions of chromosome 7 (7q-) are among the most recurrent chromosomal aberrations found in myeloid neoplasms. Patients carrying these cytogenetical alterations present a poor overall survival (OS), mainly due to a low sensitivity to standard chemotherapy and a high incidence of relapse. In our study, we aimed to disentangle the biology of patients with -7/7q- and find new candidate therapeutic targets for a disease with such a dismal prognosis, by integrating wide genomic approaches. Methods We collected bone marrow samples from 487 adult patients at diagnosis, treated in 3 institutions: Institute L. & A. Seragnoli (Italy, n = 213), CEMM (Austria, n = 160), University of Michigan (US, n = 114, GSE23452). Three hundred ninety-five samples were analyzed by SNP arrays (Affymetrix™), 51 samples by mass spectrometry (Metabolon™) and 57 samples by GEP (Affymetrix™) approaches. Chi-squared, fisher's exact test and ANOVA were used to test differences in proportion and distributions. False discovery rate, Bonferroni correction, and Welch's correction were calculated whenever appropriate. Results Among the 474 patients with evaluable karyotype, 65 (13.7%) had -7/7q-; 47 (9.9%) had -7, while 18 (3.8%) had 7q-. In our sets, the median age at AML diagnosis was 64 years (21-86) and most of the subjects had a de novo AML (65.1%). WBC count at diagnosis was significantly lower in -7/7q- patients (10.4 vs 35.2 k/mm3 p Within patients tested for FLT3, NPM1 and TP53 mutation at diagnosis, 1/50 among -7/7q- patients vs 59/300 controls harbored FLT3 ITD mutation (350 patients tested, 2% vs 19.7%, p In terms of outcome, -7/7q- AML had a median of overall survival of 10.3 months (95% C.I 5.8-14.8), which accounted for 49.5 months (95% C.I. 40.5-58.4) in other AML cases. GEP data of a cohort of 57 patients (8 with -7/7q- and 49 controls) revealed that 24 genes were under-expressed in -7/7q- AML (with e-4 significance threshold, Figure 1A). Twenty-three out of 24 genes mapped on chromosome 7; one gene, COX17, mapped on chromosome 3 (Figure 1A). COX17 plays a role in the recruitment of copper to mitochondria. By metabolomic analyses, we considered quantitative data of 300 different metabolites in 32 patients (4 with -7/7q- vs 28 controls, 19 patients were excluded because samples at diagnosis were not available). In -7/7q- AML, fatty acids (sphingomyelin, 1-linoleoylglycerol), β-cytrilglutamate and the UDP were overrepresented if compared with other AML cases; on the other hand, galactiol and 1-(1-enyl-palmitoyl)-2-linoleoyl-GPC were underrepresented in -7/7q- patients. Furthermore, -7/7q- AML cells seem to accumulate 3-hydroxy-3-methylglutarate and to have lower levels of 2-Hydroxyglutarate (Figure 1C). With SNP arrays, we considered copy number alterations in 395 patients (52 -7/7q- patients vs 343 controls, Figure 1B). 5q was the most recurrent concurrent deletion, with a minimal common deleted region (MCDR) in q31.3-q33.3. Additionally, 17p (MCDR p11.2 - p13.1), 12p (MCDR p12.3-p13.1), 16q (MCDRs q11.2-q12.1, q21-q22.1 and q24.2-q24.3), 16p (MCDR p11.2) and chromosome 4 (MCDRs q34.1 and q35.2) deletions also co-occurred in -7/7q-, listed per frequencies (Figure 1B). These regions are enriched for genes controlling cell signaling, DNA transcription, post-transcriptional modifications (such as SUMOylation), mRNA splicing and cellular senescence. Conclusions SNP, GEP and metabolomic approaches gave new insights on -7/7q- AML biology, identifying 24 new genes differentially expressed in -7/7q-, and 6 MCDR associated with -7/7q- AML. Deletion of chromosome 16q and 4 were never reported in literature associated to AML. Furthermore, for the first time, we described metabolites associated with -7/7q- AML. These data may represent a useful backbone to search for candidate targets in the setting of one of the most aggressive AML subtypes. Supported by:EHA Non-Clinical Junior Research Fellowship,HARMONY,Fondazione del Monte,FP7-NGS-PTL,AIRC. *MCF and MO equally contributed &CP and GS shared the last authorship Disclosures Haferlach: MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Kralovics:MyeloPro Diagnostics and Research GmbH: Equity Ownership. Soverini:Incyte Biosciences: Consultancy; Bristol Myers Squibb: Consultancy; Novartis: Consultancy. Martinelli:Ariad/Incyte: Consultancy; Celgene: Consultancy, Speakers Bureau; Novartis: Speakers Bureau; Pfizer: Consultancy, Speakers Bureau; Amgen: Consultancy; Janssen: Consultancy; Abbvie: Consultancy; Roche: Consultancy; Jazz Pharmaceuticals: Consultancy. Cavo:Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; GlaxoSmithKline: Honoraria, Membership on an entity's Board of Directors or advisory committees; Adaptive Biotechnologies: Honoraria, Membership on an entity's Board of Directors or advisory committees; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees; Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees; AbbVie: Honoraria, Membership on an entity's Board of Directors or advisory committees; Bristol-Myers Squibb: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau.
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- 2018
48. MDM2 and Aurora Kinase a Contribute to SETD2 Loss of Function in Advanced Systemic Mastocytosis: Implications for Pathogenesis and Treatment
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Elisa Ficarra, Cecilia Monaldi, Peter Valent, C. Avanzato, Simona Soverini, Abbenante maria Chiara, Massimiliano Bonifacio, Marianna Criscuolo, Patrizia Tosi, Roberta Zanotti, Luana Bavaro, Maria Chiara Fontana, Margherita Martelli, Chiara Elena, Sara De Santis, Cristina Papayannidis, Luigi Scaffidi, Giovanni Martinelli, Luciano Xumerle, Michele Cavo, Francesco Albano, Giulia Paciello, Fabio Ciceri, Michela Rondoni, Livio Pagano, Antonio Curti, Manuela Mancini, and Massimo Delledonne
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biology ,Bortezomib ,business.industry ,Immunology ,Cell Biology ,Hematology ,Mast cell leukemia ,medicine.disease ,Biochemistry ,Ixazomib ,Ubiquitin ligase ,chemistry.chemical_compound ,chemistry ,medicine ,biology.protein ,Cancer research ,Mdm2 ,Danusertib ,Aurora Kinase A ,Clonogenic assay ,business ,medicine.drug - Abstract
The SETD2 gene encodes the only methyltransferase responsible for histone H3 lysine 36 trimethylation (H3K36Me3) in humans. H3K36me3 play a key role in preserving the fidelity of transcription elongation and splicing. In addition, SETD2/H3K36me3 have more recently been implicated in the maintenance of genomic integrity by regulating homologous recombination (HR) repair, Mismatch Repair (MMR) mitotic spindle assembly and chromosome segregation. SETD2 deletions and/or inactivating mutations occur in many solid tumors and have recently been found also in acute leukemias. We have reported that the HMC-1.1 and -1.2 mast cell leukemia (MCL) cell lines and many advanced systemic mastocytosis (SM) patients (pts) display H3K36Me3 deficiency as a result of non-genomic loss of function of SETD2. Proteasome inhibition restored SETD2 protein expression and H3K36me3, suggesting that a functional protein is produced but rapidly degraded. In an attempt to uncover the mechanisms underlying this phenomenon, we used an in silico approach to identify candidate SETD2-interacting proteins, followed by experimental confirmation by co-immunoprecipitation (co-IP). We found that, after proteasomal inhibition, SETD2 co-immunoprecipitates with the ubiquitin E3 ligase MDM2. Treatment with the MDM2 inhibitor SP-141 rescued SETD2 expression and H3K36Me3, suggesting that MDM2 may play a role in SETD2 degradation in ASM and MCL. Moreover, SP-141 treatment of HMC-1 cells at micromolar doses induced cytostatic but not cytotoxic effects as shown by cell growth curves. Clonogenic assays supported the cytostatic effects of SP141 in HMC-1.1 and -1.2 cells. siRNA-mediated knock-down of MDM2 also rescued SETD2 expression and activity, further supporting the hypothesis that SETD2 hyper-ubiquitination by MDM2 plays a role in SETD2 reduced stability and proteasomal degradation. Co-IP also showed that SETD2 interacts with Aurora Kinase A, as it was suggested in silico. We found that Aurora A is overexpressed in advanced SM and may target SETD2 for phosphorylation. Both pharmacological inhibition by Danusertib and siRNA-mediated silencing of Aurora A rescued SETD2 expression and activity, raising the hypothesis that phosphorylation by Aurora A might be the trigger for MDM-2 mediated degradation of SETD2. To evaluate whether increased DNA damage and reduced HR proficiency can be observed in SETD2/H3K36Me3-deficient SM, we used western blotting (WB) and immunofluorescence (IF) to assess phosphorylated histone 2A.X (γH2AX) and Rad51 foci. Compared to cells from healthy controls, SETD2- and H3K36Me3-deficient cell lines and pts had significantly higher levels of γH2AX and lower levels of Rad51. RNA-seq in SETD2-deficient pts showed evidence of transcription and splicing defects like transcription-induced chimeras, intron retention and non-canonical splicing patterns not observed in healthy donors. Next, the ROSAD816V cell line, which displays SETD2 and H3K36me3 levels superimposable to healthy donors, was studied by WB and IF to assess γH2AX and Rad51 in steady state and after sub-lethal DNA damage by UV exposure. The same experiments were carried out after SETD2 silencing for 2 months. Cells with silenced SETD2 had significantly higher levels of γH2AX and were unable to activate the HR repair. Interestingly, clonogenic assays in ROSAD816V cells before and after SETD2 silencing showed that reduction of clonogenic potential after proteasomal or MDM2 inhibition is indeed SETD2-dependent (Figure 1A). Finally, we performed clonogenic assays to evaluate the therapeutic potential of bortezomib, carfilzomib and ixazomib in neoplastic mast cells from 3 patients with advanced SM and we observed in all cases that both first and second generation inhibitors induced a significant reduction of clonogenic activity at nanomolar doses (Figure 1B). Taken together, our results suggest that AKA and MDM2-mediated post-translational modifications contribute to SETD2 non-genomic loss of function in advanced SM. Loss of SETD2 and H3K36me3 is associated with increased DNA damage and transcription and splicing defects in patients. Inhibiting AKA or MDM2 activity or proteasome-mediated degradation are promising therapeutic strategies in patients with low SETD2 expression levels. Acknowledgments: study supported by AIRC (project code 16996) and AIL (Associazione Italiana contro le Leucemia, Linfomi e Mieloma). Figure 1. Figure 1. Disclosures Bonifacio: Incyte: Consultancy; Pfizer: Consultancy; Amgen: Consultancy; Novartis: Research Funding; Bristol Myers Squibb: Consultancy. Pagano:Janssen: Speakers Bureau; Merck: Speakers Bureau; Gilead: Speakers Bureau; Basilea: Speakers Bureau; Pfizer: Speakers Bureau. Valent:Novartis: Honoraria; Pfizer: Honoraria; Incyte: Honoraria. Cavo:Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Bristol-Myers Squibb: Honoraria, Membership on an entity's Board of Directors or advisory committees; AbbVie: Honoraria, Membership on an entity's Board of Directors or advisory committees; GlaxoSmithKline: Honoraria, Membership on an entity's Board of Directors or advisory committees; Adaptive Biotechnologies: Honoraria, Membership on an entity's Board of Directors or advisory committees; Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees. Soverini:Novartis: Consultancy; Incyte Biosciences: Consultancy; Bristol Myers Squibb: Consultancy.
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- 2018
49. A New Gene Expression Profile Signature CRLF2 Overexpression Based Identifies Novel Adult 'Triple Negative' Acute Lymphoblastic Leukemia Subgroups
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Antonella Padella, Silvia Vitali, Giorgia Simonetti, Daniel Remondini, Simona Righi, Alessandra Santoro, Maria Chiara Fontana, Andrea Ghelli Luserna di Rorà, Nicoletta Testoni, Eugenio Fonzi, Massimiliano Bonafè, Michele Cavo, Elena Sabattini, Anna Maria Ferrari, Giovanni Pasquini, Castellani Gastone, Giulia Ferrari, Cristina Papayannidis, Michela Tebaldi, Maria Chiara Abbenante, Valentina Robustelli, Giovanni Marconi, Samanta Salvi, Giovanni Martinelli, Enrica Imbrogno, Jesús María Hernández-Rivas, and Carmen Baldazzi
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Brachial Plexus Neuritis ,Immunology ,Cell Biology ,Hematology ,Biology ,medicine.disease ,Biochemistry ,law.invention ,Gene expression profiling ,law ,Acute lymphocytic leukemia ,Gene expression ,medicine ,Cancer research ,Immunohistochemistry ,Interleukin-7 receptor ,Burkitt's lymphoma ,Polymerase chain reaction - Abstract
Background: The heterogeneous and poor survival group of Philadelphia negative (Ph-) B-ALL patients (pts) that doesn't have the most recurrent adult rearrangements (BCR-ABL1 t(9;22); TCF3-PBX1 t(1;19); MLL-AF4 t(4;11)) are collectively referred to as "triple negative" (Ph-/-/-) ALL. CRLF2 is frequently altered in adult B-ALL, especially in Ph-like pts (50-75% of cases). Alterations that lead, in the majority of cases, to a CRLF2 overexpression. Adult pts with CRLF2 upregulated have poor outcome and novel strategies are needed to improve it. Aims: Clustering and biological characterization of Ph-/-/- ALL (that represents 61% of adult B-ALL; Roberts KG, J Clin Oncol. 2016), considering CRLF2 overexpression event, in order to define and assess biomarkers in this subgroup to test new drugs. Patients and Methods: Gene Expression Profiling (GEP; HTA 2.0 Affymetrix) were performed on 55 Ph-/-/- ALL, 29 B-ALL Ph+ at different time point of the disease and on 7 mononuclear cell of healthy donors. Data were normalized with the Expression Console Software. Successively we cluster triple negative GEP data with our validated pipeline, based on CRLF2 upregulation and in the top ten-gene list. Ph-/-/- ALL samples were then characterized for the presence of gene fusions, Copy Number Alterations (CNAs) and mutations using different approaches (TruSight Pancancer-Illumina; MLPA and/or dMLPA-MRC-Holland; SNP Array-Affymetrix; 454 Junior-Roche and PCR). Results: Clustering our Ph-/-/- gene expression data using the impact of the 10 single genes in our cohort, we could identify a defined 2-clusters-subdivision (Gr1 and Gr2; Fig 1A). The Gr2 is characterized by CTGF, CRLF2 and CD200 (Gr2=3C-up; Fig 1B) overexpression and it represents 14.1% of all B-ALL. The Gr2 GEP is similar to Ph+ one. Fusion copy number alteration and mutational screening done, detected that 3C-Up group has a higher frequency of Ph-like associated lesions (primarily CRLF2, JAK2, IL7R mutations or deletion), that mainly affect JAK-STAT pathway. Also IKZF1 and EBF1 deletions are significantly associated to Gr2 (p=0.003; p=0.016). RAS pathway genes are highly affected in Gr1. Molecular characterization shed light on a very heterogeneous scenario especially in the group 1, suggesting the need of a more discerning clustering for this group. In spite of the small number of cases is required, preliminary Gr1 subclustering discerns MLLr and ZNF384 gene expression subgroups. Notably p53 pathway is enriched in both groups but with different deregulated genes: CHEK2 is upregulated in the group1 and CDK6 in the Gr2. CRLF2 and CD200 immunoblotting and CD200 immunohistochemistry preliminary analyses suggest that protein expression of CRFL2 and CD200 are higher in Gr2 in comparison to Gr1. Conclusions: we identified a new signature, related to CRLF2 high expression, to classify Ph-/-/- ALL B-based on 10 genes. 3C-up represents 14.1% of all B-ALL and it is characterized by a) high co-expression of three main genes: CRLF2, CTGF and CD200; b) IKZF1 deletion; c) JAK-STAT pathway mutations/fusions/deletions. Gr1 represents 46.9% of all B-ALL. Gr2 GEP similarity to Ph+ one, suggests that this Gr2 could contain Ph-like pts. This new Ph-/-/- subclassification identify new potential therapeutic targets with available drug (α-CTGF, α-CD200, CDK2, CHK2 and CDK6 inhibitors; tyrosine kinase inhibitors already effective on Ph+ and Ph-like) to test. Supported by: ELN, AIL, AIRC, project Regione-Università 2010-12 (L. Bolondi), FP7 NGS-PTL project, HARMONY project, Fondazione del Monte BO e RA project. Figure. Figure. Disclosures Cavo: Bristol-Myers Squibb: Honoraria, Membership on an entity's Board of Directors or advisory committees; Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees; Adaptive Biotechnologies: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; AbbVie: Honoraria, Membership on an entity's Board of Directors or advisory committees; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees; GlaxoSmithKline: Honoraria, Membership on an entity's Board of Directors or advisory committees; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau. Martinelli:Novartis: Speakers Bureau; Abbvie: Consultancy; Jazz Pharmaceuticals: Consultancy; Janssen: Consultancy; Pfizer: Consultancy, Speakers Bureau; Roche: Consultancy; Celgene: Consultancy, Speakers Bureau; Ariad/Incyte: Consultancy; Amgen: Consultancy.
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- 2018
50. Mitoxantrone, Etoposide and Cytarabine (MEC) Can Induce Deep Complete Remission and Is an Effective Bridge Therapy to Allotransplantation (SCT) in Refractory/Relapsed Acute Myeloid Leukemia (AML) Patients
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Nicoletta Testoni, Michele Cavo, Maria Chiara Fontana, Matteo Olivi, Simone Ragaini, Carmen Baldazzi, Sarah Parisi, Maria Teresa Bochicchio, Stefano De Polo, Emanuela Ottaviani, Chiara Sartor, Giovanni Martinelli, Maria Chiara Abbenante, Luca Bertamini, Cristina Papayannidis, Stefania Paolini, Annalisa Talami, Jacopo Nanni, Antonio Curti, and Giovanni Marconi
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medicine.medical_specialty ,business.industry ,Immunology ,Cell Biology ,Hematology ,medicine.disease ,Biochemistry ,Minimal residual disease ,Chemotherapy regimen ,Helsinki declaration ,Fludarabine ,Clinical trial ,Log-rank test ,Internal medicine ,medicine ,business ,Etoposide ,Febrile neutropenia ,medicine.drug - Abstract
Introduction Relapsed/refractory (R/R) AML patients continue to be a formidable clinical challenge, mainly in consideration of associated very poor outcome, with a median overall survival (OS) of less than 12 months. SCT represents the only curative option for these patients. Although, there is no standard-of-care approach which may serve as a bridge to SCT. Our study aims to investigate the effectiveness of MEC regimen as a rescue therapy for R/R AML patients by specifically addressing the CR rate, including minimal residual disease (MRD) negativity, the number of patients who subsequently underwent SCT and the presence of predictive factors of response. Methods Fifty-five consecutive adult AML patients were treated with MEC regimen in our Institution. In patients under 66 years old, we administered mitoxantrone 6 mg/sqm/die from day 1 to day 6, etoposide 100 mg/sqm/die from day 1 to day 6 and cytarabine 1000mg/sqm/die from day 1 to day 6, whereas in patients over 66 years old, the treatment schedule was reduced to 4 consecutive days. Data were retrospectively collected by using RedCap in accordance with Helsinki declaration and GCP. We used Kaplan-Meyer to estimate survival, and log rank to test differences in survival. Chi-squared, fisher's exact test and linear-by-linear correlation were used to test differences in proportions and distributions. Response was defined in accordance with 2017 ELN recommendations. CTCAE 4.03 was used to grade adverse events. MRD was assessed with WT1 or specific fusion transcripts. Results Fifty-five patients received MEC from 2008 to 2018. Age at diagnosis ranged from 17 to 72 years, with a median age of 51 years. Our set was enriched for high-risk patients. Interestingly, twenty percent of patients harbored FLT3-ITD at diagnosis (table 1). Two main groups were included: resistant AML, 28/55 patients (50,9%), and relapsed AML, 27/55 patients (49,1%). At induction, almost half of patients received "3+7" (n=25, 45,5%), while fludarabine-based regimens were administered to 14 patients (25,5%). In our set, after MEC median duration of hospitalization was 30 days (14-78); PMN >500/mm3 was reached after 26 days (range 18-67). Fever and febrile neutropenia was the most recurrent adverse events (AE). AEs were low in grade; out of 80 graded AEs, 38 (47,5%) were grade 2, 27 (33,8%) were grade 3, 9 (11,3%) were grade 4 and only 3 events resulted in death (3,8%). E. coli was the most recurrent cause of infection (10 cases). Overall, 25/55 patients (45,5%) achieved a complete remission (CR) after one course of MEC chemotherapy. Twelve patients (21.9%) achieved MRD negativity and 13 patients (23,6%) obtained an MRD+ CR or had no MRD test. Six patients (10,9%) had a partial response (PR) and 1 patient (1,8%)had hematological improvement (HI). Four patients (7.3%) died during post-MEC aplastic phase. Disease risk at diagnosis and R/R status did not influence the chance to obtain CR (figure 1 A). In 12 patients, a second MEC was administered. Four out of 12 patient improved their response with the 2nd MEC (2 patients obtained MRD - from MRD+ CR, 1 patient obtained PR and 1 patients obtained CR from hematological improvement). MEC was an effective bridge to SCT, 32/55 patients (58,2%, figure 1 B), received SCT; 15/32 patients (46,9%) received SCT directly after the 1st course of MEC, 9/32 patients (28,1%) after the 2nd course of MEC and 2 patients (6,3%) after an additional course of post-remission chemotherapy. Of note, only 6 patients (18,8%), who were not responsive to MEC, underwent SCT after an alternative rescue therapy. Median overall survival (OS) from MEC was 455 days (95% C.I. 307-602 days.); 1-year OS, 3-year OS and 5-years OS were 57,9%, 33,2% and 23,1%, respectively (std. error ± 0,067). Patients who responded to MEC (CR MRD+ or CR MRD- after 1 or 2 courses) had better OS than non-responders (median OS 1389 vs 160 days, p=.003). Stepwise multiple logistic regression analysis with COX-HR model established that pre-MEC R/R status, diagnosis class risk, response to one or two courses of MEC, and SCT were independent predictors of survival in the optimal model. Conclusions Taken together, our data indicate that MEC is an effective salvage regimen with affordable toxicity, and gives a high chance to obtain CR. MEC is particularly useful as a bridge to SCT, and has to be considered as a rescue therapy whenever a clinical trial is not available. *GM and AT equally contributed Disclosures Martinelli: Ariad/incyte: Consultancy; Pfizer: Consultancy; Celgene: Consultancy; Amgen: Consultancy; Janssen: Consultancy; Roche: Consultancy. Cavo:Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Adaptive Biotechnologies: Honoraria, Membership on an entity's Board of Directors or advisory committees; AbbVie: Honoraria, Membership on an entity's Board of Directors or advisory committees; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees; Bristol-Myers Squibb: Honoraria, Membership on an entity's Board of Directors or advisory committees; GlaxoSmithKline: Honoraria, Membership on an entity's Board of Directors or advisory committees.
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- 2018
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