64 results on '"Lorenza Pecciarini"'
Search Results
2. Gene Fusion Detection in NSCLC Routine Clinical Practice: Targeted-NGS or FISH?
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Lorenza Pecciarini, Emanuela Brunetto, Greta Grassini, Valeria De Pascali, Francesca Rita Ogliari, Anna Talarico, Giovanna Marra, Gilda Magliacane, Miriam Redegalli, Gianluigi Arrigoni, Chiara Lazzari, Vanesa Gregorc, Alessandra Bulotta, Claudio Doglioni, and Maria Giulia Cangi more...
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fusion ,NGS ,FISH ,ALK ,ROS1 ,RET ,Cytology ,QH573-671 - Abstract
The ability to identify the broadest range of targetable gene fusions is crucial to facilitate personalized therapy selection for advanced lung adenocarcinoma (LuADs) patients harboring targetable receptor tyrosine kinase (RTK) genomic alterations. In order to evaluate the most effective testing approach for LuAD targetable gene fusion detection, we analyzed 210 NSCLC selected clinical samples, comparing in situ (Fluorescence In Situ Hybridization, FISH, and ImmunoHistoChemistry, IHC) and molecular (targeted RNA Next-Generation Sequencing, NGS, and RealTime-PCR, RT-PCR) approaches. The overall concordance among these methods was high (>90%), and targeted RNA NGS was confirmed to be the most efficient technique for gene fusion identification in clinical practice, allowing the simultaneous analysis of a large set of genomic rearrangements at the RNA level. However, we observed that FISH was useful to detect targetable fusions in those samples with inadequate tissue material for molecular testing as well as in those few cases whose fusions were not identified by the RNA NGS panel. We conclude that the targeted RNA NGS analysis of LuADs allows accurate RTK fusion detection; nevertheless, standard methods such as FISH should not be dismissed, as they can crucially contribute to the completion of the molecular characterization of LuADs and, most importantly, the identification of patients as candidates for targeted therapies. more...
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- 2023
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Catalog
3. Novel insights into the genetics and epigenetics of MALT lymphoma unveiled by next generation sequencing analyses
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Luciano Cascione, Andrea Rinaldi, Alessio Bruscaggin, Chiara Tarantelli, Alberto J. Arribas, Ivo Kwee, Lorenza Pecciarini, Afua A. Mensah, Valeria Spina, Elaine Y.L. Chung, Lodovico Terzi di Bergamo, Stephan Dirnhofer, Alexandar Tzankov, Roberto N. Miranda, Ken H. Young, Alexandra Traverse-Glehen, Gianluca Gaidano, Steven H. Swerdlow, Randy Gascoyne, Raul Rabadan, Maurilio Ponzoni, Govind Bhagat, Davide Rossi, Emanuele Zucca, and Francesco Bertoni more...
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Diseases of the blood and blood-forming organs ,RC633-647.5 - Published
- 2019
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4. Next Generation Sequencing in Non-Small Cell Lung Cancer: Pitfalls and Opportunities
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Chiara Lazzari, Alessandra Bulotta, Maria Giulia Cangi, Gabriele Bucci, Lorenza Pecciarini, Silvia Bonfiglio, Vincenza Lorusso, Stefania Ippati, Gianluigi Arrigoni, Greta Grassini, Claudio Doglioni, and Vanesa Gregorc more...
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non-small cell lung cancer ,next-generation sequencing ,molecular tumor board ,Medicine (General) ,R5-920 - Abstract
Lung cancer remains the first cause of cancer-related deaths worldwide. Thanks to the improvement in the knowledge of the biology of non-small cell lung cancer (NSCLC), patients’ survival has significantly improved. A growing number of targetable molecular alterations have been identified. Next-generation sequencing (NGS) has become one of the methodologies entered in clinical practice and was recently recommended by the European society for medical oncology (ESMO) to perform a comprehensive molecular characterization in patients with cancer. The current review provides an overview of the clinical trials that have explored the impact of NGS in patients with cancer, its limits, and advantages. more...
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- 2020
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5. Modeling multiple myeloma-bone marrow interactions and response to drugs in a 3D surrogate microenvironment
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Daniela Belloni, Silvia Heltai, Maurilio Ponzoni, Antonello Villa, Barbara Vergani, Lorenza Pecciarini, Magda Marcatti, Stefania Girlanda, Giovanni Tonon, Fabio Ciceri, Federico Caligaris-Cappio, Marina Ferrarini, and Elisabetta Ferrero more...
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Diseases of the blood and blood-forming organs ,RC633-647.5 - Abstract
Multiple myeloma develops primarily inside the bone marrow microenvironment, that confers pro-survival signals and drug resistance. 3D cultures that reproduce multiple myeloma-bone marrow interactions are needed to fully investigate multiple myeloma pathogenesis and response to drugs. To this purpose, we exploited the 3D Rotary Cell Culture System bioreactor technology for myeloma-bone marrow co-cultures in gelatin scaffolds. The model was validated with myeloma cell lines that, as assessed by histochemical and electron-microscopic analyses, engaged contacts with stromal cells and endothelial cells. Consistently, pro-survival signaling and also cell adhesion-mediated drug resistance were significantly higher in 3D than in 2D parallel co-cultures. The contribution of the VLA-4/VCAM1 pathway to resistance to bortezomib was modeled by the use of VCAM1 transfectants. Soluble factor-mediated drug resistance could be also demonstrated in both 2D and 3D co-cultures. The system was then successfully applied to co-cultures of primary myeloma cells-primary myeloma bone marrow stromal cells from patients and endothelial cells, allowing the development of functional myeloma-stroma interactions and MM cell long-term survival. Significantly, genomic analysis performed in a high-risk myeloma patient demonstrated that culture in bioreactor paralleled the expansion of the clone that ultimately dominated in vivo. Finally, the impact of bortezomib on myeloma cells and on specialized functions of the microenvironment could be evaluated. Our findings indicate that 3D dynamic culture of reconstructed human multiple myeloma microenvironments in bioreactor may represent a useful platform for drug testing and for studying tumor-stroma molecular interactions. more...
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- 2018
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6. CDC25A Protein Stability Represents a Previously Unrecognized Target of HER2 Signaling in Human Breast Cancer: Implication for a Potential Clinical Relevance in Trastuzumab Treatment
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Emanuela Brunetto, Anna Maria Ferrara, Francesca Rampoldi, Anna Talarico, Elena DalCin, Greta Grassini, Lorenzo Spagnuolo, Isabella Sassi, Antonella Ferro, Lucia Veronica Cuorvo, Mattia Barbareschi, Sara Piccinin, Roberta Maestro, Lorenza Pecciarini, Claudio Doglioni, and Maria Giulia Cangi more...
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Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
The CDC25A-CDK2 pathway has been proposed as critical for the oncogenic action of human epidermal growth factor receptor 2 (HER2) in mammary epithelial cells. In particular, transgenic expression of CDC25A cooperates with HER2 in promoting mammary tumors, whereas CDC25A hemizygous loss attenuates the HER2-induced tumorigenesis penetrance. On the basis of this evidence of a synergism between HER2 and the cell cycle regulator CDC25A in a mouse model of mammary tumorigenesis, we investigated the role of CDC25A in human HER2-positive breast cancer and its possible implications in therapeutic response. HER2 status and CDC25A expression were assessed in 313 breast cancer patients and we found statistically significant correlation between HER2 and CDC25A (P = .007). Moreover, an HER2-positive breast cancer subgroup with high levels of CDC25A and very aggressive phenotype was identified (P = .005). Importantly, our in vitro studies on breast cancer cell lines showed that the HER2 inhibitor efficacy on cell growth and viability relied also on CDC25A expression and that such inhibition induces CDC25A down-regulation through phosphatidylinositol 3-kinase/protein kinase B pathway and DNA damage response activation. In line with this observation, we found a statistical significant association between CDC25A overexpression and trastuzumab-combined therapy response rate in two different HER2-positive cohorts of trastuzumab-treated patients in either metastatic or neoadjuvant setting (P = .018 for the metastatic cohort and P = .021 for the neoadjuvant cohort). Our findings highlight a link between HER2 and CDC25A that positively modulates HER2- targeted therapy response, suggesting that, in HER2-positive breast cancer patients, CDC25A overexpression affects trastuzumab sensitivity. more...
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- 2013
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7. Correction: Establishment and Characterization of PCL12, a Novel CD5+ Chronic Lymphocytic Leukaemia Cell Line.
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Andreas Agathangelidis, Lydia Scarfò, Federica Barbaglio, Benedetta Apollonio, Maria Teresa Sabrina Bertilaccio, Pamela Ranghetti, Maurilio Ponzoni, Gabriella Leone, Valeria De Pascali, Lorenza Pecciarini, Paolo Ghia, Federico Caligaris-Cappio, and Cristina Scielzo more...
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Medicine ,Science - Published
- 2015
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8. Establishment and Characterization of PCL12, a Novel CD5+ Chronic Lymphocytic Leukaemia Cell Line.
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Andreas Agathangelidis, Lydia Scarfò, Federica Barbaglio, Benedetta Apollonio, Maria Teresa Sabrina Bertilaccio, Pamela Ranghetti, Maurilio Ponzoni, Gabriella Leone, Valeria De Pascali, Lorenza Pecciarini, Paolo Ghia, Federico Caligaris-Cappio, and Cristina Scielzo more...
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Medicine ,Science - Abstract
Immortalized cell lines representative of chronic lymphocytic leukemia (CLL) can assist in understanding disease pathogenesis and testing new therapeutic agents. At present, very few representative cell lines are available. We here describe the characterization of a new cell line (PCL12) that grew spontaneously from the peripheral blood (PB) of a CLL patient with progressive disease and EBV infection. The CLL cell origin of PCL12 was confirmed after the alignment of its IGH sequence against the "original" clonotypic sequence. The IGH gene rearrangement was truly unmutated and no CLL-related cytogenetic or genetic lesions were detected. PCL12 cells express CD19, CD20, CD5, CD23, low levels of IgM and IgD and the poor-outcome-associated prognostic markers CD38, ZAP70 and TCL1. In accordance with its aggressive phenotype the cell line is inactive in terms of LYN and HS1 phosphorylation. BcR signalling pathway is constitutively active and anergic in terms of p-ERK and Calcium flux response to α-IgM stimulation. PCL12 cells strongly migrate in vitro in response to SDF-1 and form clusters. Finally, they grow rapidly and localize in all lymphoid organs when xenotrasplanted in Rag2-/-γc-/- mice. PCL12 represents a suitable preclinical model for testing pharmacological agents. more...
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- 2015
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9. Routine Molecular Profiling in Both Resectable and Unresectable Pancreatic Adenocarcinoma: Relevance of Cytologic Samples
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Miriam Redegalli, Greta Grassini, Gilda Magliacane, Lorenza Pecciarini, Marco Schiavo Lena, Chanel E. Smart, Rebecca L. Johnston, Nicola Waddell, Roberta Maestro, Marina Macchini, Giulia Orsi, Maria Chiara Petrone, Gemma Rossi, Gianpaolo Balzano, Massimo Falconi, Paolo G. Arcidiacono, Michele Reni, Claudio Doglioni, and Maria Giulia Cangi more...
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Hepatology ,Gastroenterology - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive disease, for which it is crucial to promptly detect actionable and prognostic alterations to drive specific therapeutic decisions, regardless of tumor resectability status. Endoscopic ultrasonography-guided fine-needle aspiration (EUS-FNA) is of key importance for PDAC diagnosis and can contribute significantly to tumor molecular profiling.Comprehensive genomic profile by targeted next-generation sequencing (NGS) was performed on 2 independent PDAC patient cohorts. Cohort 1 consisted of 77 patients with resectable PDAC for whom the histologic sample at the time of resection was available; for 56 patients cytologic specimens at the time of diagnosis also were obtained by EUS-FNA. Cohort 2 consisted of 20 patients with unresectable PDAC, for whom only the EUS-FNA cytologic sample was available.In cohort 1, a complete concordant mutational profile between the cytologic sample at diagnosis and the corresponding histologic specimen after surgery was observed in 88% of the cases, proving the ability to detect potential clinically relevant alterations in cytologic samples by NGS analysis. Notably, clinically actionable mutations were identified in 20% of patients. In cohort 2, comprehensive mutational profiling was obtained successfully for all samples. Consistent with the findings of cohort 1, KRAS, TP53, CDKN2A, and SMAD4 were the most altered genes. Most importantly, 15% of the patients harbored actionable mutations.Our findings show the feasibility of an NGS approach using both surgical specimens and cytologic samples. The model proposed in this study can be included successfully in the clinical setting for comprehensive molecular profiling of all PDAC patients irrespective of their surgical eligibility. more...
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- 2022
10. Safety and efficacy of a dose-dense short-term therapy in patients with MYC-translocated aggressive lymphoma
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Andrés J. M. Ferreri, Piera Angelillo, Federico Erbella, Chiara Cattaneo, Luisa Verga, Arben Lleshi, Bernardino Allione, Maurilio Ponzoni, Fabio Facchetti, Chiara Pagani, Marco Foppoli, Lorenza Pecciarini, Marianna Sassone, Sara Steffanoni, Elena Flospergher, Giuseppe Rossi, Michele Spina, and Alessandro Re more...
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Lymphoma, B-Cell ,Lymphoma ,Hematopoietic Stem Cell Transplantation ,Cytarabine ,COVID-19 ,HIV Infections ,Hematology ,Transplantation, Autologous ,Burkitt Lymphoma ,Antibodies, Monoclonal, Murine-Derived ,Vincristine ,Doxorubicin ,Antineoplastic Combined Chemotherapy Protocols ,Humans ,Prednisone ,Rituximab ,Cyclophosphamide ,In Situ Hybridization, Fluorescence ,Etoposide ,Retrospective Studies - Abstract
Patients with aggressive B-cell lymphoma and MYC rearrangement at fluorescence in situ hybridization exhibit poor outcome after R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, prednisone). In the last decade, 68 patients with Burkitt lymphoma ([BL] n = 46) or high-grade B-cell lymphoma ([HGBCL] single, double, or triple hit; n = 22) were treated with a dose-dense, short-term therapy termed “CARMEN regimen” at 5 Italian centers. Forty-six (68%) patients were HIV+. CARMEN included a 36-day induction with sequential, single weekly doses of cyclophosphamide, vincristine, rituximab, methotrexate, etoposide, and doxorubicin plus intrathecal chemotherapy, followed by high-dose-cytarabine–based consolidation. Patients who did not achieve complete remission (CR) after induction received BEAM (carmustina, etoposide, cytarabine, melfalan)-conditioned autologous stem cell transplantation (ASCT) after consolidation. Sixty-one (90%) patients completed induction, and 59 (87%) completed consolidation. Seventeen patients received ASCT. Grade 4 hematological toxicity was common but did not cause treatment discontinuation; grade 4 nonhematological toxicity was recorded in 11 (16%) patients, with grade 4 infections in 6 (9%). Six (9%) patients died of toxicity (sepsis in 4, COVID-19, acute respiratory distress syndrome). CR rate after the whole treatment was 73% (95% confidence interval [CI], 55% to 91%) for patients with HGBCL and 78% (95% CI, 66% to 90%) for patients with BL. At a median follow-up of 65 (interquartile range, 40-109) months, 48 patients remain event free, with a 5-year progression-free survival of 63% (95% CI, 58% to 68%) for HGBCL and 72% (95% CI, 71% to 73%) for BL, with a 5-year overall survival (OS) of 63% (95% CI, 58% to 68%) and 76% (95% CI, 75% to 77%), respectively. HIV seropositivity did not have a detrimental effect on outcome. This retrospective study shows that CARMEN is a safe and active regimen both in HIV-negative and -positive patients with MYC-rearranged lymphomas. Encouraging survival figures, attained with a single dose of doxorubicin and cyclophosphamide, deserve further investigation in HGBCL and other aggressive lymphomas. more...
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- 2022
11. Locally Performed HRD Testing for Ovarian Cancer? Yes, We Can!
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Gilda Magliacane, Emanuela Brunetto, Silvia Calzavara, Alice Bergamini, Giovanni Battista Pipitone, Giovanna Marra, Miriam Redegalli, Greta Grassini, Emanuela Rabaiotti, Gianluca Taccagni, Lorenza Pecciarini, Paola Carrera, Giorgia Mangili, Claudio Doglioni, and Maria Giulia Cangi more...
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Cancer Research ,Oncology ,BRCA ,HRD ,HRR ,NGS ,molecular testing ,ovarian cancer ,PARPi - Abstract
Assessment of HRD status is now essential for ovarian cancer patient management. A relevant percentage of high-grade serous carcinoma (HGSC) is characterized by HRD, which is caused by genetic alterations in the homologous recombination repair (HRR) pathway. Recent trials have shown that not only patients with pathogenic/likely pathogenic BRCA variants, but also BRCAwt/HRD patients, are sensitive to PARPis and platinum therapy. The most common HRD test is Myriad MyChoice CDx, but there is a pressing need to offer an alternative to outsourcing analysis, which typically requires high costs and lengthy turnaround times. In order to set up a complete in-house workflow for HRD testing, we analyzed a small cohort of HGSC patients using the CE-IVD AmoyDx HRD Focus Panel and compared our results with Myriad’s. In addition, to further deepen the mechanisms behind HRD, we analyzed the study cohort by using both a custom NGS panel that analyzed 21 HRR-related genes and FISH analysis to determine the copy numbers of PTEN and EMSY. We found complete concordance in HRD status detected by the Amoy and the Myriad assays, supporting the feasibility of internal HRD testing. more...
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- 2022
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12. Intratumoral cellular heterogeneity: Implications for drug resistance in patients with non-small cell lung cancer
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Maria Grazia Viganò, Vanesa Gregorc, Francesca Rita Ogliari, Greta Grassini, Lorenza Pecciarini, Aurora Mirabile, Gianluigi Arrigoni, Alessandra Bulotta, Maria Giulia Cangi, Claudio Doglioni, Chiara Lazzari, Abdelrahman Khater, Stefania Ippati, Mario Mandalà, Giulia Veronesi, Gregorc, V., Lazzari, C., Mandala, M., Ippati, S., Bulotta, A., Cangi, M. G., Khater, A., Vigano, M. G., Mirabile, A., Pecciarini, L., Ogliari, F. R., Arrigoni, G., Grassini, G., Veronesi, G., and Doglioni, C. more...
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0301 basic medicine ,Oncology ,Genome instability ,Genomic instability ,Cancer Research ,medicine.medical_specialty ,Tumor heterogeneity ,Review ,Drug resistance ,NSCLC ,03 medical and health sciences ,0302 clinical medicine ,Non-small cell lung cancer ,Internal medicine ,medicine ,In patient ,Epigenetics ,Lung cancer ,RC254-282 ,Targeted agents ,business.industry ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,medicine.disease ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer cell ,Non small cell ,business - Abstract
Simple Summary The number of druggable tumor-specific molecular alterations in the treatment of non-small cell lung cancer (NSCLC) has grown significantly in the past decade. Emerging technologies such as liquid biopsy and single-cell methods allow for studying targetable drivers and develop personalized treatments. However, although new therapies confer prolonged disease control and high tumor response rates, most patients eventually progress on targeted treatments. Intratumoral heterogeneity is a frequent event in NSCLC, driving the tumor cells to develop adaptive or new resistance mechanisms within the drug environment. This review summarizes the current and upcoming research on the biological role of tumor heterogeneity, highlighting the link between early and acquired drug resistance and tumoral heterogeneity in targetable driver mutated NSCLC. Abstract Tailored therapies based on the identification of molecular targets currently represent a well-established therapeutic scenario in the treatment of non-small cell lung cancer (NSCLC) patients. However, while aiming to improve patients’ response to therapy, development of resistance is frequently observed in daily clinical practice. Intratumoral heterogeneity is a frequent event in NSCLC, responsible for several critical issues in patients’ diagnosis and treatment. Advances in single-cell sequencing technologies have allowed in-depth profiling of tumors and attributed intratumoral heterogeneity to genetic, epigenetic, and protein modification driven diversities within cancer cell populations. This review highlights current research on the biological role of tumor heterogeneity and its impact on the development of acquired resistance in NSCLC patients. more...
- Published
- 2021
13. Next Generation Sequencing in Non-Small Cell Lung Cancer: Pitfalls and Opportunities
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Greta Grassini, Vanesa Gregorc, Lorenza Pecciarini, Stefania Ippati, Chiara Lazzari, Claudio Doglioni, Silvia Bonfiglio, Alessandra Bulotta, Vincenza Lorusso, Maria Giulia Cangi, Gabriele Bucci, and Gianluigi Arrigoni more...
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0301 basic medicine ,Oncology ,medicine.medical_specialty ,Clinical Biochemistry ,Review ,molecular tumor board ,DNA sequencing ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,medicine ,In patient ,Lung cancer ,non-small cell lung cancer ,lcsh:R5-920 ,business.industry ,Cancer ,medicine.disease ,Clinical Practice ,Clinical trial ,030104 developmental biology ,030220 oncology & carcinogenesis ,next-generation sequencing ,Non small cell ,business ,lcsh:Medicine (General) - Abstract
Lung cancer remains the first cause of cancer-related deaths worldwide. Thanks to the improvement in the knowledge of the biology of non-small cell lung cancer (NSCLC), patients’ survival has significantly improved. A growing number of targetable molecular alterations have been identified. Next-generation sequencing (NGS) has become one of the methodologies entered in clinical practice and was recently recommended by the European society for medical oncology (ESMO) to perform a comprehensive molecular characterization in patients with cancer. The current review provides an overview of the clinical trials that have explored the impact of NGS in patients with cancer, its limits, and advantages. more...
- Published
- 2020
14. Acute Lung injury evolution in Covid-19
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Giulio Rossi, Antonella Arcadu, Vanni Agnoletti, Luca Donati, Federica Pedica, Alessandra Dubini, Claudio Doglioni, Marco Chilosi, Elisabetta Fabbri, Vittorio Sambri, Silvia Puglisi, Claudia Ravaglia, Venerino Poletti, Carmela Grosso, Sara Piciucchi, Stefano Maitan, Vincenzo Bronte, Stefano Ugel, Giovanni Pizzolo, Simona di Cesare, Lorenza Pecciarini, Athol U. Wells, Emanuele Russo, Antonio Vizzuso, Giovanni Poletti, Franco Stella, and Emiliano Gamberini more...
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Pathology ,medicine.medical_specialty ,Lung ,business.industry ,Microangiopathy ,Lung biopsy ,Hyperplasia ,Lung injury ,medicine.disease ,Pathogenesis ,medicine.anatomical_structure ,medicine ,Cytokine storm ,business ,Diffuse alveolar damage - Abstract
BackgroundPathogenesis of Coronavirus disease 2019 (Covid-19) is poorly understood. Most histologic studies come from post-mortem analysis, with existing data indicating that histologic features of acute respiratory distress syndrome are typically present in fatal cases. However, this observation may be misleading, due to confounding factors in pre-terminal disease, including injury resulting from prolonged mechanical ventilation. Ante-mortem lung biopsy may provide major pathogenetic insights, potentially providing a basis for novel treatment approaches.AimThis comparative, multicenter, prospective, observational study was planned to identify ante-mortem histological profile and immunohistochemical features of lung tissue in patients with Covid-19 in early and late phases of the disease, including markers of inflammatory cells and major pathways involved in the cytokine storm triggering.MethodsEnrolled patients underwent lung biopsy, according to the study protocol approved by local Ethical Committee, either within 15 days of the first symptoms appearing (early phase) or after >15 days (more advanced disease). Key exclusion criteria were excessive or uncorrectable bleeding risk and cardiovascular disease with heart failure. Lung samples were obtained by conventional transbronchial biopsy, trans-bronchial lung cryobiopsy or surgical lung biopsy.Results23 patients were enrolled: 12 patients underwent lung biopsy within 15 days and 11 patients more than 15 days after the onset of symptoms. Early biopsies were characterized by spots of patchy acute lung injury (ALI) with alveolar type II cells hyperplasia and significant vascular abnormalities (disordered angiogenesis with alveolar capillary hyperplasia, luminal enlargement and thickened walls of pulmonary venules, perivascular CD4-T-cell infiltration), with no hyaline membranes. In the later stages, the alveolar architecture appeared disrupted, with areas of organizing ALI, venular congestion and capillary thromboembolic microangiopathy. Striking phenotypic features were demonstrated in hyperplastic pneumocytes and endothelial cells, including the expression of phospho-STAT3 and molecules involved in immunoinhibitory signals (PD-L1 and IDO-1). Alveolar macrophages exhibited macrophage-related markers (CD68, CD11c, CD14) together with unusual markers, such as DC-Lamp/CD208, CD206, CD123/IL3AR.ConclusionA morphologically distinct “Covid pattern” was identified in the earlier stages of the disease, with prominent epithelial and endothelial cell abnormalities, that may be potentially reversible, differing strikingly from findings in classical diffuse alveolar damage. These observations may have major therapeutic implications, justifying studies of early interventions aimed at mitigating inflammatory organ injury. more...
- Published
- 2020
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15. Prospective Validation of the Italian Alliance Against Cancer Lung Panel in Patients With Advanced Non-Small-Cell Lung Cancer
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Laura Bonanno, Concetta Di Micco, Alessandro Guida, Lorenza Pecciarini, Gianmaria Frigè, Gabriele Bucci, Arnaud Ceol, Simona Coco, Arianna Marinello, Carlo Genova, Pier Giuseppe Pelicci, Maria Giulia Cangi, Gennaro Ciliberto, Paolo Graziano, Simonetta Buglioni, Alessandro Bandiera, Angelo Delmonte, Chiara Lazzari, Silvia Bonfiglio, Paolo Vigneri, Luca Toschi, Gianmarco Motta, Ruggero De Maria, Luca Mazzarella, Antonio Rossi, and Vanesa Gregorc more...
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0301 basic medicine ,Pulmonary and Respiratory Medicine ,Oncology ,Cancer Research ,medicine.medical_specialty ,bioinformatics ,molecular tumor board ,next-generation sequencing ,NSCLC ,personalized therapy ,Lung Neoplasms ,Bioinformatics ,Personalized therapy ,Molecular tumor board ,Sensitivity and Specificity ,03 medical and health sciences ,Therapeutic approach ,Next-generation sequencing ,Carcinoma, Non-Small-Cell Lung ,Early Detection of Cancer ,Genomics ,Humans ,Italy ,Mass Screening ,Precision Medicine ,Prospective Studies ,0302 clinical medicine ,Settore MED/04 - PATOLOGIA GENERALE ,Internal medicine ,Medicine ,In patient ,Lung cancer ,Non-Small-Cell Lung ,Lung ,Molecular screening ,business.industry ,Carcinoma ,Cancer ,medicine.disease ,Precision medicine ,respiratory tract diseases ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Non small cell ,business - Abstract
Background The deeper knowledge of non–small-cell lung cancer (NSCLC) biology and the discovery of driver molecular alterations have opened the era of precision medicine in lung oncology, thus significantly revolutionizing the diagnostic and therapeutic approach to NSCLC. In Italy, however, molecular assessment remains heterogeneous across the country, and numbers of patients accessing personalized treatments remain relatively low. Nationwide programs have demonstrated that the creation of consortia represent a successful strategy to increase the number of patients with a molecular classification. Patients and Methods The Alliance Against Cancer (ACC), a network of 25 Italian Research Institutes, has developed a targeted sequencing panel for the detection of genomic alterations in 182 genes in patients with a diagnosis of NSCLC (ACC lung panel). One thousand metastatic NSCLC patients will be enrolled onto a prospective trial designed to measure the sensitivity and specificity of the ACC lung panel as a tool for molecular screening compared to standard methods. Results and Conclusion The ongoing trial is part of a nationwide strategy of ACC to develop infrastructures and improve competences to make the Italian research institutes independent for genomic profiling of cancer patients. more...
- Published
- 2020
16. Acute Lung injury evolution in Covid-19
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Claudio, Doglioni, primary, Claudia, Ravaglia, additional, Giulio, Rossi, additional, Alessandra, Dubini, additional, Federica, Pedica, additional, Sara, Piciucchi, additional, Antonio, Vizzuso, additional, Lorenza, Pecciarini, additional, Franco, Stella, additional, Stefano, Maitan, additional, Vanni, Agnoletti, additional, Emiliano, Gamberini, additional, Emanuele, Russo, additional, Silvia, Puglisi, additional, Antonella, Arcadu, additional, Luca, Donati, additional, Simona, Di Cesare, additional, Carmela, Grosso, additional, Giovanni, Poletti, additional, Vittorio, Sambri, additional, Elisabetta, Fabbri D, additional, Giovanni, Pizzolo, additional, Stefano, Ugel, additional, Vincenzo, Bronte, additional, Athol, Wells U, additional, Marco, Chilosi, additional, and Venerino, Poletti, additional more...
- Published
- 2020
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17. A dose-dense short-term therapy for human immunodeficiency virus/acquired immunodeficiency syndrome patients with high-risk Burkitt lymphoma or high-grade B-cell lymphoma: safety and efficacy results of the 'CARMEN' phase II trial
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Michele Spina, Maurilio Ponzoni, Luigi Rigacci, Bernardino Allione, Alessandro Re, Marco Foppoli, Andrés J.M. Ferreri, Marianna Sassone, Lorenza Pecciarini, Giovanni Donadoni, Luca Fumagalli, Chiara Cattaneo, Teresa Calimeri, Fabio Facchetti, Arben Lleshi, Giuseppe Rossi, Luisa Verga, and Daris Ferrari more...
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Adult ,Male ,Antimetabolites, Antineoplastic ,medicine.medical_specialty ,Lymphoma, B-Cell ,HIV Infections ,MYC ,central nervous system prophylaxis ,Antiviral Agents ,Transplantation, Autologous ,Gastroenterology ,03 medical and health sciences ,0302 clinical medicine ,Autologous stem-cell transplantation ,Interquartile range ,Internal medicine ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Mucositis ,Humans ,Melphalan ,Etoposide ,high-grade B-cell lymphoma ,Acquired Immunodeficiency Syndrome ,Meningeal Lymphoma ,business.industry ,Cytarabine ,Hematopoietic Stem Cell Transplantation ,Burkitt lymphoma ,Hematology ,Middle Aged ,medicine.disease ,Carmustine ,Lymphoma ,Tolerability ,030220 oncology & carcinogenesis ,double-hit lymphoma ,Human Immunodeficiency Virus ,Female ,business ,030215 immunology ,medicine.drug - Abstract
A few prospective trials in HIV-positive patients with Burkitt lymphoma (BL) or high-grade B-cell lymphoma (HGBL) have been reported. Investigated therapies have shown good efficacy but relevant safety problems, with high rates of interruptions, severe mucositis, septic complications, and fungal infections. Here, we report the results of a multicentre phase II trial addressing a new dose-dense, short-term therapy aimed at maintaining efficacy and improving tolerability. The experimental programme included a 36-day polychemotherapy induction followed by high-dose cytarabine-based consolidation and response-tailored BEAM (carmustine, etoposide, cyatarabine, and melphalan)- conditioned autologous stem cell transplantation (ASCT). This therapy would be considered active if ≥11 complete remissions (CR) after induction (primary endpoint) were recorded among 20 assessable patients. HIV-positive adults (median age 42, range 26-58; 16 males) with untreated BL (n = 16), HGBL (n = 3) or double-hit lymphoma (n = 1) were enrolled. All patients had high-risk features, with meningeal and bone marrow infiltration in five and nine patients respectively. The experimental programme was safe and active in a multicentre setting, with only two episodes of grade 4 non-haematological toxicity (hepatotoxicity and mucositis), and no cases of systemic fungal infections; two patients died of toxicity (bacterial infections). Response after induction (median duration: 47 days; interquartile range 41-54), was complete in 13 patients and partial in five [overall response rate = 90%; 95% confidence interval (CI) = 77-100]. All responders received consolidation, and five required autologous stem cell transplant. At a median follow-up of 55 (41-89) months, 14 patients are relapse-free and 15 are alive, with a five-year progression-free survival and an overall survival of 70% (95% CI = 60-80%) and 75% (95% CI = 66-84) respectively. No patient with cerebrospinal fluid (CSF)/meningeal lymphoma experienced central nervous system recurrence. With respect to previously reported regimens, this programme was delivered in a shorter period, and achieved the main goal of maintaining efficacy and improving tolerability. more...
- Published
- 2020
18. Evidence of a common cell origin in a case of pancreatic mixed intraductal papillary mucinous neoplasm–neuroendocrine tumor
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Lorenza Pecciarini, Greta Grassini, Claudio Doglioni, Renaud Maire, Marco Schiavo Lena, Aurel Perren, Maria Giulia Cangi, Ilaria Francaviglia, Massimo Falconi, Stefano Partelli, Schiavo Lena, M., Cangi, M. G., Pecciarini, L., Francaviglia, I., Grassini, G., Maire, R., Partelli, S., Falconi, M., Perren, A., and Doglioni, C. more...
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0301 basic medicine ,Male ,Pathology ,medicine.medical_specialty ,KRAS and GNAS mutation ,Adenoma ,Cell ,DNA Mutational Analysis ,Pancreatic Intraductal Neoplasms ,medicine.disease_cause ,Pathology and Forensic Medicine ,03 medical and health sciences ,0302 clinical medicine ,CDKN2A ,Pancreatic neuroendocrine tumor ,medicine ,GNAS complex locus ,Biomarkers, Tumor ,Humans ,Cyclin D1 amplification ,610 Medicine & health ,Molecular Biology ,biology ,Intraductal papillary mucinous neoplasm ,Precursor lesion ,Mixed neuroendocrine non-neuroendocrine neoplasms ,Cell Biology ,General Medicine ,Middle Aged ,medicine.disease ,Adenocarcinoma, Mucinous ,CDKN2A mutation ,Carcinoma, Papillary ,Pancreatic Neoplasms ,stomatognathic diseases ,Neuroendocrine Tumors ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,biology.protein ,570 Life sciences ,KRAS ,Who classification ,Carcinoma, Pancreatic Ductal - Abstract
Recently, the term mixed neuroendocrine non-neuroendocrine neoplasms (MiNEN) has been proposed as an umbrella definition covering different possible combinations of mixed neuroendocrine-exocrine neoplasms. Among these, the adenoma plus neuroendocrine tumor (NET) combination is among the rarest and not formally recognized by the 2019 WHO Classification. In this setting, the debate between either collision tumors or true mixed neoplasms is still unsolved. In this report, a pancreatic intraductal papillary mucinous neoplasm (IPMN) plus a NET is described, and the molecular investigations showed the presence in both populations of the same KRAS, GNAS, and CDKN2A mutations and the amplification of the CCND1 gene. These data prove clonality and support a common origin of both components, therefore confirming the true mixed nature. For this reason, mixed neuroendocrine-exocrine neoplasms, in which the exocrine component is represented by a glandular precursor lesion (adenoma/IPMN) only, should be included into the MiNEN family. more...
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- 2020
19. Identification and monitoring of somatic mutations in circulating cell-free tumor DNA in lung cancer patients
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Emanuela Brunetto, Greta Grassini, Chiara Lazzari, Lorenza Pecciarini, Ilaria Francaviglia, Vanesa Gregorc, Daniela Medicina, Alessandra Bulotta, Elena Dal Cin, Maria Giulia Cangi, Salvatore Girlando, Claudio Doglioni, Chanel Smart, Gilda Magliacane, Francaviglia, I., Magliacane, G., Lazzari, C., Grassini, G., Brunetto, E., Dal Cin, E., Girlando, S., Medicina, D., Smart, C. E., Bulotta, A., Gregorc, V., Pecciarini, L., Doglioni, C., and Cangi, M. G. more...
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0301 basic medicine ,Pulmonary and Respiratory Medicine ,Oncology ,Adult ,Male ,Cancer Research ,medicine.medical_specialty ,Lung Neoplasms ,Drug resistance ,Target therapy ,Molecular diagnostic ,Circulating Tumor DNA ,03 medical and health sciences ,T790M ,0302 clinical medicine ,Antineoplastic Agents, Immunological ,Internal medicine ,Positron Emission Tomography Computed Tomography ,Biopsy ,medicine ,Biomarkers, Tumor ,Humans ,Molecular Targeted Therapy ,Lung cancer ,Lung ,Protein Kinase Inhibitors ,In Situ Hybridization, Fluorescence ,Aged ,Aged, 80 and over ,Massive parallel sequencing ,medicine.diagnostic_test ,business.industry ,Liquid Biopsy ,High-Throughput Nucleotide Sequencing ,ctDNA ,DNA, Neoplasm ,Middle Aged ,medicine.disease ,Molecular diagnostics ,Resistance mutation ,030104 developmental biology ,NGS ,030220 oncology & carcinogenesis ,Mutation ,Female ,EGFR Activating Mutation ,business - Abstract
Objectives Circulating cell-free tumor DNA (ctDNA) isolated from the peripheral blood of non-small-cell lung cancer (NSCLC) patients provides biomarkers for both therapeutic target selection, particularly when direct tumor biopsy is unfeasible, and also for drug resistance monitoring. This study evaluates the reliability and feasibility of ctDNA analysis in an in-house clinical molecular diagnostic workflow. Materials and methods Mutation profiling by both standard methods and Next-Generation sequencing (NGS) was carried out and compared on 2 independent lung cancer patient cohorts. Cohort 1 consisted of 50 EGFR-mutated NSCLC patients, established on tumour biopsy, for whom ctDNA was collected at disease progression after TKI-inhibitor treatment and could be used to monitor drug resistance. Cohort 2 consisted of 50 newly diagnosed lung cancer patients for whom tumour biopsy was not possible and only ctDNA was available, providing the possibility of biomarker identification. Results ctDNA analysis of Cohort 1 verified the persistence of the tumour-detected EGFR activating mutation at disease progression by both standard and NGS methods, in 84% and 92% of the cases respectively. The T790M EGFR resistance mutation was identified in 71% of the ctDNA EGFR mutated samples providing vital information for their disease management. In newly diagnosed Cohort 2 patients, EGFR activating mutations were detected in 16% of the patients by both standard and NGS analysis of ctDNA in peripheral blood, providing indication to targeted-therapy otherwise unavailable for this group of patients. Conclusion The presented study investigated lung cancer ctDNA analysis, comparing conventional methods versus NGS sequencing, and demonstrated the successful use of plasma ctDNA as a template for targeted NGS tumor gene panel in an in-house routine clinical practice. More importantly, these data underline the advantages of the clinical application of ctDNA NGS analysis for identification of therapeutic targets, real-time monitoring of therapy, and resistance mechanisms in lung cancer patients. more...
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- 2019
20. MYD88 L265P MUTATION DETECTION IN THE AQUEOUS HUMOR OF PATIENTS WITH VITREORETINAL LYMPHOMA
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Chiara Giuffrè, Elisabetta Miserocchi, Francesco Bandello, Maurilio Ponzoni, Teresa Calimeri, Giulio Modorati, Lorenza Pecciarini, Andrés J.M. Ferreri, Ilaria Francaviglia, Maria Giulia Cangi, Miserocchi, E., Ferreri, A. J. M., Giuffre, C., Cangi, M. G., Francaviglia, I., Calimeri, T., Ponzoni, M., Pecciarini, L., Bandello, F. M., and Modorati, G. M. more...
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Male ,genetic structures ,medicine.medical_treatment ,DNA Mutational Analysis ,Vitrectomy ,Polymerase Chain Reaction ,anterior chamber paracentesis ,law.invention ,0302 clinical medicine ,Intraocular Lymphoma ,law ,Paracentesis ,Medicine ,Prospective Studies ,Prospective cohort study ,Polymerase chain reaction ,Aged, 80 and over ,medicine.diagnostic_test ,Primary central nervous system lymphoma ,vitreoretinal lymphoma ,General Medicine ,DNA, Neoplasm ,Middle Aged ,MYD88 L265P mutation ,030220 oncology & carcinogenesis ,Mutation (genetic algorithm) ,Female ,Waldenstrom Macroglobulinemia ,medicine.medical_specialty ,Aqueous humor ,Slit Lamp Microscopy ,Aqueous Humor ,03 medical and health sciences ,Ophthalmology ,Biomarkers, Tumor ,Humans ,Aged ,primary central nervous system lymphoma ,business.industry ,medicine.disease ,eye diseases ,Vitreous Body ,Mutation ,Myeloid Differentiation Factor 88 ,030221 ophthalmology & optometry ,sense organs ,business ,Vitreoretinal lymphoma - Abstract
Purpose To detect the presence of MYD88 L265P mutation in the aqueous humor of patients with cytologically proven vitreoretinal lymphoma. Methods Eight consecutive patients with bilateral vitreoretinal lymphoma (16 eyes) were prospectively evaluated. Genomic DNA was extracted from aqueous samples after paracentesis and vitreous humor samples after diagnostic vitrectomy. MYD88 codon 265 mutation was investigated by both amplification-refractory mutation system polymerase chain reaction approach and pyrosequencing assay in the aqueous humor of all patients and in the vitreous of 6 patients. A control group of 8 age-matched patients with established diagnosis of noninfectious uveitis was also tested for the presence of MYD88 L265P mutation in the aqueous humor. Results Eight patients (three men, five women) with mean age of 69.5 years (range 50-85 years) were considered. All the patients tested for MYD88 L265P in the vitreous (six) were positive, and this result was consistent with cytological examination in all samples but one. The MYD88 L265P mutation was found in the aqueous of 6 patients (75%), and in 3 of them, the mutation was present in both eyes. Results of MYD88 L265P mutation in aqueous and vitreous sample were consistent in 7 of the 8 eyes with available samples. The aqueous humor of the noninfectious uveitis control group was negative for the detection of MYD88 L265P mutation. Conclusion MYD88 mutation was detected in the aqueous humor of 75% of patients with cytologically proven vitreoretinal lymphoma. This technique may be considered as an additional diagnostic tool in the detection of the disease. more...
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- 2018
21. Rapid targeted somatic mutation analysis of solid tumors in routine clinical diagnostics
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Paola Bartocci, Gianluca Barbieri, Gilda Magliacane, Claudio Doglioni, Gianluigi Arrigoni, Greta Grassini, Lorenza Pecciarini, Maria Giulia Cangi, Elena Dal Cin, Ilaria Francaviglia, Magliacane, Gilda, Grassini, Greta, Bartocci, Paola, Francaviglia, Ilaria, Dal Cin, Elena, Barbieri, Gianluca, Arrigoni, Gianluigi, Pecciarini, Lorenza, Doglioni, Claudio, and Cangi Maria, Giulia more...
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Neuroblastoma RAS viral oncogene homolog ,Proto-Oncogene Proteins B-raf ,Genotyping Techniques ,Class I Phosphatidylinositol 3-Kinases ,medicine.medical_treatment ,DNA Mutational Analysis ,medicine.disease_cause ,Bioinformatics ,Polymorphism, Single Nucleotide ,Targeted therapy ,GTP Phosphohydrolases ,molecular diagnostics ,Proto-Oncogene Proteins p21(ras) ,symbols.namesake ,Phosphatidylinositol 3-Kinases ,Neoplasms ,medicine ,Pathology ,Humans ,Genotyping ,Retrospective Studies ,Sanger sequencing ,Base Sequence ,business.industry ,target therapy ,Cancer ,High-Throughput Nucleotide Sequencing ,Membrane Proteins ,Reproducibility of Results ,solid tumors ,Molecular diagnostics ,medicine.disease ,mutations ,Research Paper: Pathology ,ErbB Receptors ,Oncology ,Molecular Diagnostic Techniques ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Mutation ,symbols ,high throughput mass-spectrometry ,KRAS ,business - Abstract
// Gilda Magliacane 1, * , Greta Grassini 1, * , Paola Bartocci 2, * , Ilaria Francaviglia 1 , Elena Dal Cin 1 , Gianluca Barbieri 2 , Gianluigi Arrigoni 1 , Lorenza Pecciarini 1 , Claudio Doglioni 1 , Maria Giulia Cangi 1 1 Unit of Pathology, IRCCS San Raffaele Scientific Institute, Milano, Italy 2 Diatech Pharmacogenetics Company, Jesi, Italy * These authors have contributed equally to this work Correspondence to: Maria Giulia Cangi, e-mail: cangi.mariagiulia@hsr.it Claudio Doglioni, e-mail: doglioni.claudio@hsr.it Keywords: Pathology, molecular diagnostics, solid tumors, high throughput mass-spectrometry, target therapy, mutations Received: July 01, 2015 Accepted: August 14, 2015 Published: September 02, 2015 ABSTRACT Tumor genotyping is an essential step in routine clinical practice and pathology laboratories face a major challenge in being able to provide rapid, sensitive and updated molecular tests. We developed a novel mass spectrometry multiplexed genotyping platform named PentaPanel to concurrently assess single nucleotide polymorphisms in 56 hotspots of the 5 most clinically relevant cancer genes, KRAS, NRAS, BRAF , EGFR and PIK3CA for a total of 221 detectable mutations. To both evaluate and validate the PentaPanel performance, we investigated 1025 tumor specimens of 6 different cancer types (carcinomas of colon, lung, breast, pancreas, and biliary tract, and melanomas), systematically addressing sensitivity, specificity, and reproducibility of our platform. Sanger sequencing was also performed for all the study samples. Our data showed that PentaPanel is a high throughput and robust tool, allowing genotyping for targeted therapy selection of 10 patients in the same run, with a practical turnaround time of 2 working days. Importantly, it was successfully used to interrogate different DNAs isolated from routinely processed specimens (formalin-fixed paraffin embedded, frozen, and cytological samples), covering all the requirements of clinical tests. In conclusion, the PentaPanel platform can provide an immediate, accurate and cost effective multiplex approach for clinically relevant gene mutation analysis in many solid tumors and its utility across many diseases can be particularly relevant in multiple clinical trials, including the new basket trial approach, aiming to identify appropriate targeted drug combination strategies. more...
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- 2015
22. Modeling multiple myeloma-bone marrow interactions and response to drugs in a 3d surrogate microenvironment
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Stefania Girlanda, Marina Ferrarini, Lorenza Pecciarini, Elisabetta Ferrero, Federico Caligaris-Cappio, Magda Marcatti, Giovanni Tonon, Maurilio Ponzoni, Antonello Villa, Silvia Heltai, Barbara Vergani, Fabio Ciceri, Daniela Belloni, Belloni, D, Heltai, S, Ponzoni, M, Villa, A, Vergani, B, Pecciarini, L, Marcatti, M, Girlanda, S, Tonon, G, Ciceri, F, Caligaris-Cappio, F, Ferrarini, M, Ferrero, E, Belloni, Daniela, Heltai, Silvia, Ponzoni, Maurilio, Villa, Antonello, Vergani, Barbara, Pecciarini, Lorenza, Marcatti, Magda, Girlanda, Stefania, Tonon, Giovanni, Ciceri, Fabio, Caligaris-Cappio, Federico, Ferrarini, Marina, and Ferrero, Elisabetta more...
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0301 basic medicine ,3D model ,medicine.medical_specialty ,Stromal cell ,Cell Survival ,Cell Culture Techniques ,Drug Resistance ,Cell Communication ,Models, Biological ,Article ,Plasma Cell Disorders ,Bortezomib ,03 medical and health sciences ,Bioreactors ,0302 clinical medicine ,Bone Marrow ,immune system diseases ,hemic and lymphatic diseases ,Internal medicine ,Cell Adhesion ,Tumor Microenvironment ,medicine ,Humans ,Multiple myeloma ,Tumor microenvironment ,Hematology ,Chemistry ,Endothelial Cells ,medicine.disease ,Coculture Techniques ,Bone Marrow Microenvironment ,030104 developmental biology ,medicine.anatomical_structure ,Cell culture ,030220 oncology & carcinogenesis ,Cancer research ,Gelatin ,Bone marrow ,Stromal Cells ,Clone (B-cell biology) ,Multiple Myeloma ,medicine.drug - Abstract
Multiple myeloma develops primarily inside the bone marrow microenvironment, that confers pro-survival signals and drug resistance. 3D cultures that reproduce multiple myeloma-bone marrow interactions are needed to fully investigate multiple myeloma pathogenesis and response to drugs. To this purpose, we exploited the 3D Rotary Cell Culture System bioreactor technology for myeloma-bone marrow co-cultures in gelatin scaffolds. The model was validated with myeloma cell lines that, as assessed by histochemical and electron-microscopic analyses, engaged contacts with stromal cells and endothelial cells. Consistently, pro-survival signaling and also cell adhesion-mediated drug resistance were significantly higher in 3D than in 2D parallel co-cultures. The contribution of the VLA-4/VCAM1 pathway to resistance to bortezomib was modeled by the use of VCAM1 transfectants. Soluble factor-mediated drug resistance could be also demonstrated in both 2D and 3D co-cultures. The system was then successfully applied to co-cultures of primary myeloma cells-primary myeloma bone marrow stromal cells from patients and endothelial cells, allowing the development of functional myeloma-stroma interactions and MM cell long-term survival. Significantly, genomic analysis performed in a high-risk myeloma patient demonstrated that culture in bioreactor paralleled the expansion of the clone that ultimately dominated in vivo. Finally, the impact of bortezomib on myeloma cells and on specialized functions of the microenvironment could be evaluated. Our findings indicate that 3D dynamic culture of reconstructed human multiple myeloma microenvironments in bioreactor may represent a useful platform for drug testing and for studying tumor-stroma molecular interactions. more...
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- 2018
23. Establishment, characterization and long-term culture of human endocrine pancreas-derived microvascular endothelial cells
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Augusto Pessina, Emilio Ciusani, Lorenza Pecciarini, Giulio Alessandri, Anna Ferri, Rita Nano, Silvia Pellegrini, Lorenzo Piemonti, Erica Dugnani, Luisa Pascucci, Valeria Sordi, Valentina Ceserani, Sordi, V., Ferri, A., Ceserani, V., Ciusani, E., Dugnani, E., Pellegrini, S., Nano, R., Pecciarini, L., Pessina, A., Pascucci, L., Piemonti, Lorenzo, Alessandri, G., and Pathology/molecular and cellular medicine more...
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0301 basic medicine ,CD31 ,Vascular Endothelial Growth Factor A ,Cancer Research ,medicine.medical_specialty ,Endothelium ,Angiogenesis ,endothelial cell line ,islets ,pancreas ,primary culture ,Immunology ,Biology ,03 medical and health sciences ,chemistry.chemical_compound ,Islets of Langerhans ,Antigens, CD ,Internal medicine ,von Willebrand Factor ,Journal Article ,medicine ,Immunology and Allergy ,Humans ,Genetics (clinical) ,Cells, Cultured ,Transplantation ,Vascular Endothelial Growth Factor Receptor-1 ,Research Support, Non-U.S. Gov't ,Pancreatic islets ,Interleukin-8 ,Endothelial Cells ,Cell Biology ,Cadherins ,Cell biology ,Vascular endothelial growth factor ,Endothelial stem cell ,Vascular endothelial growth factor A ,030104 developmental biology ,Endocrinology ,medicine.anatomical_structure ,Oncology ,Vascular endothelial growth factor C ,chemistry ,Microvessels ,Endothelium, Vascular - Abstract
Background In vitro primary cultures of microvascular endothelial cells from endocrine pancreas are difficult to obtain, but can be a very helpful tool for studies of islet biology, transplantation and regenerative medicine. Methods We applied a protocol recently described for the isolation and culture of brain microvascular endothelial cells (EC) on human pancreatic islets. EC obtained were characterized in terms of morphological (light and transmission electron microscopy), phenotypical (by immunofluorescence and flow cytometry) and functional (cord formation assay and protein secretion by multiplex bead-based assay) characteristics. Results EC were obtained from 25% of islet preparations processed. Two primary endothelial cell lines showed high proliferative potential and were deeply characterized: they presented endothelial cell morphology and expressed CD31, CD49a, CD49e, CD34, von Willebrand Factor (vWF), Vascular Endothelial CAdherin (VE-CAD), Tyrosine Kinase with Ig and EGF Homology Domains-2 (TIE2), Vascular Endothelial Growth Factor Receptor 1 (VEGFR1), Ulex lectin and the endothelium endocrine-specific marker nephrin. Besides, they were able to form cordons in vitro and secreted factors involved in the process of angiogenesis such as Vascular Endothelial Growth Factor (VEGF), Monocyte Chemotactic Protein 1 (MCP-1), interleukin (IL)-8 and Melanoma Growth Stimulatory Activity Alpha (GROα). These cell lines were termed Human Islet Microvascular Endothelial Cells (HIMEC). Discussion This study establishes a simple and effective strategy for isolation and long-term culture of EC derived from human pancreatic islet. HIMEC in culture preserve phenotype and functional properties and are, therefore, a useful tool for future experiments of in vitro pancreas modelling, co-transplantation with pancreatic islets, re-vascularization of scaffold or matrix for regenerative medicine purposes. © 2017 International Society for Cellular Therapy more...
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- 2016
24. General population low-count CLL-like MBL persists over time without clinical progression, although carrying the same cytogenetic abnormalities of CLL
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Antonis Dagklis, Lorenza Pecciarini, Agnieszka Janus, Francesca Cottini, Paolo Ghia, Cinzia Sala, Daniela Toniolo, Lydia Scarfò, Federico Caligaris-Cappio, Cristina Scielzo, Claudia Fazi, Anna Talarico, Fazi, C, Scarfò, L, Pecciarini, L, Cottini, F, Dagklis, A, Janus, A, Talarico, A, Scielzo, C, Sala, C, Toniolo, D, CALIGARIS CAPPIO, Federico, and Ghia, PAOLO PROSPERO more...
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Adult ,Male ,Lymphocytosis ,Receptors, Antigen, T-Cell, alpha-beta ,T-Lymphocytes ,Chronic lymphocytic leukemia ,Immunology ,Population ,chemical and pharmacologic phenomena ,Biology ,Biochemistry ,immune system diseases ,hemic and lymphatic diseases ,medicine ,Humans ,Lymphocyte Count ,education ,In Situ Hybridization, Fluorescence ,Aged ,Aged, 80 and over ,Chromosome Aberrations ,B-Lymphocytes ,education.field_of_study ,Chromosomes, Human, Pair 12 ,Chromosomes, Human, Pair 13 ,Cell Biology ,Hematology ,Middle Aged ,Flow Cytometry ,bacterial infections and mycoses ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,V(D)J Recombination ,Leukemia ,Monoclonal ,Disease Progression ,Chromosome abnormality ,Monoclonal B-cell lymphocytosis ,Female ,medicine.symptom ,CD5 ,Chromosomes, Human, Pair 17 ,Follow-Up Studies - Abstract
Monoclonal B-cell lymphocytosis (MBL) is classified as chronic lymphocytic leukemia (CLL)–like, atypical CLL, and CD5− MBL. The number of B cells per microliter divides CLL-like MBL into MBL associated with lymphocytosis (usually detected in a clinical setting) and low-count MBL detected in the general population (usually identified during population screening). After a median follow-up of 34 months we reevaluated 76 low-count MBLs with 5-color flow cytometry: 90% of CLL-like MBL but only 44.4% atypical CLL and 66.7% CD5− MBL persisted over time. Population-screening CLL-like MBL had no relevant cell count change, and none developed an overt leukemia. In 50% of the cases FISH showed CLL-related chromosomal abnormalities, including monoallelic or biallelic 13q deletions (43.8%), trisomy 12 (1 case), and 17p deletions (2 cases). The analysis of the T-cell receptor β (TRBV) chains repertoire showed the presence of monoclonal T-cell clones, especially among CD4highCD8low, CD8highCD4low T cells. TRBV2 and TRBV8 were the most frequently expressed genes. This study indicates that (1) the risk of progression into CLL for low-count population-screening CLL-like MBL is exceedingly rare and definitely lower than that of clinical MBL and (2) chromosomal abnormalities occur early in the natural history and are possibly associated with the appearance of the typical phenotype. more...
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- 2011
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25. Abstract 645: Targeted next generation sequencing in molecular diagnostics of solid tumors
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Maria Giulia Cangi, Greta Grassini, Lorenza Pecciarini, Valeria De Pascali, Chiara Iacona, Elena Dal Cin, Anna Talarico, Claudio Doglioni, Ilaria Francaviglia, and Gilda Magliacane
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Cancer Research ,Molecular pathology ,Cancer ,Genomics ,Computational biology ,Biology ,Molecular diagnostics ,medicine.disease ,medicine.disease_cause ,DNA sequencing ,Oncology ,medicine ,ROS1 ,Copy-number variation ,KRAS - Abstract
Cancer genomics role in patients management underlines the importance of rigorous validation of molecular diagnostics approaches to guide treatment decisions. Nowadays only next generation sequencing (NGS) assays can satisfy the need to match a comprehensive tumor molecular profile to the right therapies. Nevertheless, given the complexity, cost, and outcome implications for patients, a clinically valid NGS assay requires careful consideration of gene panel size, inclusion of appropriate markers, ability to detect multiple genomic aberration types, performance with low quality/quantity of nucleic acids, and workflow feasibility. In this study we describe the introduction of the Oncomine Comprehensive Assay (OCA) into a Molecular Pathology diagnostic framework. The OCA analyzes 143 genes of which 73 oncogenes are interrogated for mutational hotspots and 26 tumor suppressor genes for all exons. In addition, the OCA detects copy number variations in 49 genes and fusion drivers in 22 genes. Multiplexed PCR-based DNA and RNA libraries were sequenced on Ion S5 and data analysis was performed by the Torrent Suite using the Oncomine pipeline. The OCA was clinically validated analyzing a cohort of 200 tumor samples of 4 solid tumor types (100 Non Small Cell Lung Cancers, 40 Colon Adenocarcinomas, 40 Pancreatic Adenocarcinomas, 20 Esophageal Adenocarcinomas), for which routine molecular analysis was available. We successfully identified relevant somatic point mutations, indels, and high-level CNAs: TP53, KRAS and EGFR were the genes most frequently mutated though alteration frequencies varied among different tumor types, as expected. OCA DNA profiling demonstrated 95% concordance with standard methods for detecting commonly targeted somatic variants with highly concordant observed variant allele frequencies for detection of KRAS, EGFR and BRAF mutations. Also high-level CNAs strongly correlated with available routine standard tests. The concordance of OCA analysis and standard tests at the RNA level was definitely lower (80%): a subset of 45 lung samples were tested for ALK, RET, ROS1 rearrangements by OCA RNA assay and we successfully detected 15 of the 24 expected fusions, indicating the feasibility of identifying them by this NGS panel, but also stressing the limits of a targeted approach to detect widely variable breakpoints in the analyzed gene fusions. In order to detect all the ALK, ROS1, and RET fusions variants, we perform the parallel validation of the Archer FusionPlex RNA-based ALK, RET, and ROS1 gene fusion assay in 8 lung adenocarcinomas. Results are under evaluation. We conclude that the OCA DNA analysis can be a valuable tool in solid tumor molecular diagnostics being able to identify both common and additional relevant variants beyond current routine practice, and represents a broadly applicable targeted NGS assay which allows advanced precision oncology for different solid tumors types. Citation Format: Greta Grassini, Valeria De Pascali, Ilaria Francaviglia, Gilda Magliacane, Elena Dal Cin, Anna Talarico, Chiara Iacona, Claudio Doglioni, Lorenza Pecciarini, Maria Giulia Cangi. Targeted next generation sequencing in molecular diagnostics of solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 645. more...
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- 2018
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26. Abstract 5580: Identification and monitoring of somatic mutations in circulating cell-free DNA by next-generation sequencing in patients with lung adenocarcinoma
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Alessandra Bulotta, Claudio Doglioni, Ilaria Francaviglia, Gilda Magliacane, Daniela Medicina, Greta Grassini, Salvatore Girlando, Lorenza Pecciarini, Maria Giulia Cangi, and Chiara Lazzari
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Neuroblastoma RAS viral oncogene homolog ,Oncology ,Cancer Research ,medicine.medical_specialty ,business.industry ,medicine.disease ,medicine.disease_cause ,Real-time polymerase chain reaction ,Internal medicine ,ROS1 ,Medicine ,Adenocarcinoma ,Digital polymerase chain reaction ,KRAS ,business ,Lung cancer ,Genotyping - Abstract
The analysis of circulating cell-free tumor DNA (ctDNA), which can be obtained from plasma by non-invasive procedures, was proven useful to provide biomarkers in the management of non-small-cell lung cancer (NSCLC) patients. Several studies assessed ctDNA prognostic and predictive value as source of key data for therapeutic target selection and drug resistance in such patients. The purpose of the present study was to compare the assessment of NSCLC common hot spot mutations in ctDNA samples using the NGS Oncomine cfDNA Lung Cancer assay (ThermoFisher) to the standard clinical tests (i.e. real-time quantitative PCR and droplet digital PCR), performed in both FFPE tumor and ctDNA samples. In particular we aimed to evaluate the feasibility and accuracy of this assay for (1) the detection of EGFR/KRAS mutations in 50 patients with newly diagnosed lung cancer and no possibility to obtain tissue samples and (2) the identification of anti-EGFR treatment acquired resistance mutations in 50 NSCLC EGFR-mutated patients with disease progression. The Oncomine cfDNA Lung assay is a multiplexed sequencing test, designed to detect 150 hotspot mutations in 11 lung cancer-related genes (ALK, BRAF, EGFR, ERBB2, KRAS, MAP2K1, MET, NRAS, PIK3CA, ROS1, and TP53) with a limit of detection as low as 0.1%. We analyzed a total of 100 ctDNAs which were sequenced using the Ion S5 system. Libraries were templated using the Ion Chef and multiplexed as 8 libraries on a 520 chip. Data analysis was performed in Torrent Suite using the variant Caller plugin. The total process time, from ctDNA isolation to result reporting, was as short as 4 days, supporting a workflow where blood samples are received early on day 1 and final variant calls are available on day 4. ctDNA NGS analysis for the newly diagnosed patients with no available tumor sample identified KRAS mutations (7%) and more importantly, targetable EGFR mutations (10%). For the patients with progression disease, EGFR acquired resistance mutations were found in 78% of the cases. Overall, the most frequently mutated genes were EGFR (85%), TP53 (26%), PIK3CA (18%), ALK (10%), and MET (8%). Taken the EGFR mutation detected by routine methods as the gold standard, the concordance with EGFR variants detected by NGS was 97%. Interestingly, the NGS approach detected the same EGFR mutations with the same allelic frequency of standard methods. These preliminary data confirm the potential of the Oncomine cfDNA lung assay for plasma genotyping which allows both noninvasive multiplexed detection of targetable genomic alterations and monitoring of acquired resistance mutations in lung cancer. Citation Format: Ilaria Francaviglia, Gilda Magliacane, Greta Grassini, Salvatore Girlando, Daniela Medicina, Chiara Lazzari, Alessandra Bulotta, Lorenza Pecciarini, Claudio Doglioni, Maria Giulia Cangi. Identification and monitoring of somatic mutations in circulating cell-free DNA by next-generation sequencing in patients with lung adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5580. more...
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- 2018
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27. Chlamydophila psittaciis viable and infectious in the conjunctiva and peripheral blood of patients with ocular adnexal lymphoma: Results of a single‐center prospective case–control study
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Antonio Giordano Resti, S. Magnino, Lorenza Pecciarini, Riccardo Dolcetti, Lucia Malabarba, Maria Giulia Cangi, Maurilio Ponzoni, Elisa Pasini, Nadia Vicari, G.P. Dognini, Andrés J.M. Ferreri, Silvano Rossini, Claudio Doglioni, Ferreri, Ajm, Dolcetti, R, Dognini, Gp, Malabarba, L, Vicari, N, Pasini, E, Ponzoni, Maurilio, Cangi, Mg, Pecciarini, L, Resti, Ag, Doglioni, Claudio, Rossini, S, and Magnino, S. more...
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Adult ,DNA, Bacterial ,Male ,Conjunctival Neoplasm ,Cancer Research ,Conjunctiva ,Conjunctival Neoplasms ,Polymerase Chain Reaction ,Peripheral blood mononuclear cell ,Ocular Adnexal Lymphoma ,Risk Factors ,Occupational Exposure ,medicine ,Humans ,Prospective Studies ,Animal Husbandry ,Aged ,Aged, 80 and over ,Chlamydia psittaci ,Infectivity ,biology ,business.industry ,MALT lymphoma ,Environmental Exposure ,Lymphoma, B-Cell, Marginal Zone ,Environmental exposure ,Chlamydia Infections ,Middle Aged ,Conjunctivitis ,biology.organism_classification ,medicine.disease ,medicine.anatomical_structure ,Chlamydophila psittaci ,Oncology ,Case-Control Studies ,Chronic Disease ,Immunology ,Leukocytes, Mononuclear ,Orbital Neoplasms ,Female ,business - Abstract
Ocular adnexal MALT lymphoma (OAML) is linked to Chlamydophila psittaci (Cp) infection. Viability and infectivity of Cp, demonstrated by growth in culture, has not been yet investigated in these patients. We conducted a single-center prospective case-control study to assess the prevalence, viability and infectivity of Cp in 20 OAML patients and 42 blood donors registered in a 6-month period. The presence of Cp in conjunctival swabs and peripheral blood mononuclear cells (PBMC) of patients and donors was assessed by TETR-PCR and in vitro cultures. From an epidemiological point of view, OAML patients often resided in rural areas, and reported a history of chronic conjunctivitis and prolonged contact with household animals (85% vs. 38% of donors; p = 0.00001). Cp was detected in lymphoma tissue in 15 (75%) patients. Cp DNA was detected in conjunctival swabs and/or PBMC from 10 (50%) patients and in PBMC from 1 (2%) donor (p = 0.01). Viability and infectivity of Cp, demonstrated by growth in culture, were confirmed in conjunctival swabs and/or PBMC from 5 (25%) patients, but not in donors (p = 0.002). This prospective study demonstrates, for the first time, that Cp present in the conjunctiva and PBMC of OAML patients is capable to grow and be isolated in cell cultures. Cp infection is common in OAML patients and exceptional in blood donors. Epidemiological data of OAML patients (prolonged contact with household animals and chronic conjunctivitis) are consistent with Cp exposure risk. (C) 2008 Wiley-Liss, Inc. Ocular adnexal MALT lymphoma (OAML) is linked to Chlamydophila psittaci (Cp) infection. Viability and infectivity of Cp, demonstrated by growth in culture, has not been yet investigated in these patients. We conducted a single-center prospective case-control study to assess the prevalence, viability and infectivity of Cp in 20 OAML patients and 42 blood donors registered in a 6-month period. The presence of Cp in conjunctival swabs and peripheral blood mononuclear cells (PBMC) of patients and donors was assessed by TETR-PCR and in vitro cultures. From an epidemiological point of view, OAML patients often resided in rural areas, and reported a history of chronic conjunctivitis and prolonged contact with household animals (85% vs. 38% of donors; p = 0.00001). Cp was detected in lymphoma tissue in 15 (75%) patients. Cp DNA was detected in conjunctival swabs and/or PBMC from 10 (50%) patients and in PBMC from 1 (2%) donor (p = 0.01). Viability and infectivity of Cp, demonstrated by growth in culture, were confirmed in conjunctival swabs and/or PBMC from 5 (25%) patients, but not in donors (p = 0.002). This prospective study demonstrates, for the first time, that Cp present in the conjunctiva and PBMC of OAML patients is capable to grow and be isolated in cell cultures. Cp infection is common in OAML patients and exceptional in blood donors. Epidemiological data of OAML patients (prolonged contact with household animals and chronic conjunctivitis) are consistent with Cp exposure risk. (C) 2008 Wiley-Liss, Inc. more...
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- 2008
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28. Human malignant mesothelioma is recapitulated in immunocompetent BALB/c mice injected with murine AB cells
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Hilda de Vries, Roberta Frapolli, Elena Gatti, Camilla Recordati, Michela Frenquelli, Maurizio D'Incalci, Marco Bianchi, Massimo Venturini, Antonello E. Spinelli, Francesco De Marchis, Luca Crippa, Renzo Boldorini, Eltjona Rrapaj, Rosanna Mezzapelle, Eugenio Scanziani, Claudio Doglioni, Massimo P. Crippa, Chiara Ceriotti, Laura Perani, Silvia Valtorta, Lorenza Pecciarini, Alessandro Preti, Rosa Maria Moresco, Anton Berns, Mezzapelle, R, Rrapaj, E, Gatti, E, Ceriotti, C, De Marchis, F, Preti, A, Spinelli, A, Perani, L, Venturini, M, Valtorta, S, Moresco, R, Pecciarini, L, Doglioni, C, Frenquelli, M, Crippa, L, Recordati, C, Scanziani, E, De Vries, H, Berns, A, Frapolli, R, Boldorini, R, D'Incalci, M, Bianchi, M, Crippa, M, Spinelli, Ae, Moresco, Rm, Doglioni, Claudio, de Vries, H, D’Incalci, M, Bianchi, MARCO EMILIO, and Crippa, Mp more...
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Mesothelioma ,0301 basic medicine ,Pathology ,medicine.medical_specialty ,Lung Neoplasms ,Antineoplastic Agents ,Pemetrexed ,HMGB1 ,Deoxycytidine ,Article ,Proinflammatory cytokine ,BALB/c ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Malignant Mesotheliom ,Cell Line, Tumor ,medicine ,Mesothelioma, HMGB1, in vivo, imaging ,cancer ,Animals ,Humans ,HMGB1 Protein ,Cisplatin ,Mice, Inbred BALB C ,Multidisciplinary ,biology ,business.industry ,Mesothelioma, Malignant ,biology.organism_classification ,medicine.disease ,Survival Analysis ,Gemcitabine ,030104 developmental biology ,Cell culture ,mesothelioma ,030220 oncology & carcinogenesis ,biology.protein ,Female ,business ,Immunocompetence ,Neoplasm Transplantation ,medicine.drug - Abstract
Malignant Mesothelioma is a highly aggressive cancer, which is difficult to diagnose and treat. Here we describe the molecular, cellular and morphological characterization of a syngeneic system consisting of murine AB1, AB12 and AB22 mesothelioma cells injected in immunocompetent BALB/c mice, which allows the study of the interplay of tumor cells with the immune system. Murine mesothelioma cells, like human ones, respond to exogenous High Mobility Group Box 1 protein, a Damage-Associated Molecular Pattern that acts as a chemoattractant for leukocytes and as a proinflammatory mediator. The tumors derived from AB cells are morphologically and histologically similar to human MM tumors, and respond to treatments used for MM patients. Our system largely recapitulates human mesothelioma, and we advocate its use for the study of MM development and treatment. Malignant Mesothelioma is a highly aggressive cancer, which is difficult to diagnose and treat. Here we describe the molecular, cellular and morphological characterization of a syngeneic system consisting of murine AB1, AB12 and AB22 mesothelioma cells injected in immunocompetent BALB/c mice, which allows the study of the interplay of tumor cells with the immune system. Murine mesothelioma cells, like human ones, respond to exogenous High Mobility Group Box 1 protein, a Damage-Associated Molecular Pattern that acts as a chemoattractant for leukocytes and as a proinflammatory mediator. The tumors derived from AB cells are morphologically and histologically similar to human MM tumors, and respond to treatments used for MM patients. Our system largely recapitulates human mesothelioma, and we advocate its use for the study of MM development and treatment more...
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- 2016
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29. Characterization of t(6;11)(p21;q12) in a renal-cell carcinoma of an adult patient
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Claudio Doglioni, Ettore Macrì, Crocifissa Lo Cunsolo, Lorenza Pecciarini, Guido Martignoni, Elena Dal Cin, M. Giulia Cangi, Pecciarini, L, Cangi, Mg, LO CUNSOLO, C, Macri', E, DAL CIN, E, Martignoni, G, and Doglioni, Claudio more...
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Cancer Research ,Molecular Sequence Data ,Chromosomal translocation ,Biology ,Polymerase Chain Reaction ,Translocation, Genetic ,Fusion gene ,Renal cell carcinoma ,Genetics ,medicine ,Carcinoma ,Humans ,Amino Acid Sequence ,Carcinoma, Renal Cell ,In Situ Hybridization, Fluorescence ,DNA Primers ,Base Sequence ,Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ,Chromosomes, Human, Pair 11 ,Helix-Loop-Helix Motifs ,breakpoint cluster region ,Chromosome Mapping ,Chromosome ,Middle Aged ,medicine.disease ,Molecular biology ,Fusion protein ,Kidney Neoplasms ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,Karyotyping ,TFEB ,Chromosomes, Human, Pair 6 ,Female ,Gene Fusion - Abstract
Renal-cell carcinoma (RCC) constitutes a heterogeneous group of tumors with specific chromosome aberrations. Recently, a new small group of RCC, occurring in children and young adults, has been described as characterized by t(6;11)(p21;q12). It has been shown that this translocation results in the fusion of the 5′ portion of the ALPHA gene (11q12) with the transcription factor gene TFEB (6p21). Herewith, we report the first complete cytogenetic and molecular characterization of a t(6;11)-positive RCC of an adult patient, a 54-year-old woman. The tumor was histologically defined as RCC with peculiar features and it was negative for epithelial markers and positive for melanocytic markers. Chromosome QFQ banding analysis of short-term cultured cells from the RCC showed t(6;11)(p21;q12) as the sole cytogenetic abnormality. The translocation was confirmed by FISH analysis. RT-PCR analysis, performed on total RNA isolated from both neoplastic and normal tissue samples, revealed an ALPHA–TFEB chimeric transcript in the tumor sample; sequencing of the RT-PCR product defined a novel TFEB gene breakpoint cluster region, broader than the one reported thus far. Western blot analysis showed a band at the expected size of wild-type TFEB in the neoplastic tissue compared to the normal sample, supporting that the fusion gene does not encode for a chimeric protein but it causes an upregulation of the wild-type TFEB. Our data contribute to define better this rare RCC type, which is typical not only of childhood but can also be found in adulthood. © 2007 Wiley-Liss, Inc. more...
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- 2007
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30. Correction: Establishment and Characterization of PCL12, a Novel CD5+ Chronic Lymphocytic Leukaemia Cell Line
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Paolo Ghia, Gabriella Leone, Lorenza Pecciarini, Lydia Scarfò, Maurilio Ponzoni, Andreas Agathangelidis, Cristina Scielzo, Maria Teresa Sabrina Bertilaccio, Federico Caligaris-Cappio, Pamela Ranghetti, Valeria De Pascali, Federica Barbaglio, and Benedetta Apollonio more...
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Gene Rearrangement ,Multidisciplinary ,Lymphocytic leukaemia ,ZAP-70 Protein-Tyrosine Kinase ,business.industry ,Science ,Correction ,CD5 Antigens ,Leukemia, Lymphocytic, Chronic, B-Cell ,Mice ,Phenotype ,Cell culture ,Cell Line, Tumor ,Cancer research ,Medicine ,Animals ,Humans ,CD5 ,business ,Neoplasm Transplantation - Abstract
Immortalized cell lines representative of chronic lymphocytic leukemia (CLL) can assist in understanding disease pathogenesis and testing new therapeutic agents. At present, very few representative cell lines are available. We here describe the characterization of a new cell line (PCL12) that grew spontaneously from the peripheral blood (PB) of a CLL patient with progressive disease and EBV infection. The CLL cell origin of PCL12 was confirmed after the alignment of its IGH sequence against the "original" clonotypic sequence. The IGH gene rearrangement was truly unmutated and no CLL-related cytogenetic or genetic lesions were detected. PCL12 cells express CD19, CD20, CD5, CD23, low levels of IgM and IgD and the poor-outcome-associated prognostic markers CD38, ZAP70 and TCL1. In accordance with its aggressive phenotype the cell line is inactive in terms of LYN and HS1 phosphorylation. BcR signalling pathway is constitutively active and anergic in terms of p-ERK and Calcium flux response to α-IgM stimulation. PCL12 cells strongly migrate in vitro in response to SDF-1 and form clusters. Finally, they grow rapidly and localize in all lymphoid organs when xenotrasplanted in Rag2-/-γc-/- mice. PCL12 represents a suitable preclinical model for testing pharmacological agents. more...
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- 2015
31. Alterations of β-Catenin Pathway in Non-Melanoma Skin Tumors
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Sara Piccinin, Michela Armellin, Lorenza Pecciarini, Concetta Chiarelli, Roberta Maestro, Maria Giulia Cangi, Silvia Demontis, Claudio Doglioni, Tamara Vukosavljevic, and Mauro Boiocchi
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Mutation ,Pathology ,medicine.medical_specialty ,biology ,Adenomatous polyposis coli ,Wnt signaling pathway ,Gene mutation ,medicine.disease_cause ,medicine.disease ,Pathology and Forensic Medicine ,Catenin ,Trichoepithelioma ,biology.protein ,medicine ,Cancer research ,AXIN2 ,Carcinogenesis - Abstract
To determine the role of β-catenin pathway in human skin carcinogenesis, 135 non-melanoma skin tumors were analyzed for β-catenin expression and gene mutations. Intense nucleo-cytoplasmic immunoreactivity for C terminus β-catenin antibodies was observed in all pilomatricomas and in single cases of trichoepithelioma and squamous cell carcinoma showing peculiar signs of matrical differentiation. Moderate increase of β-catenin nuclear staining was detected in a significant proportion of basal cell carcinomas, Bowen disease, spiroadenomas, and occasionally also in squamous cell carcinomas, but in these neoplasms only a limited fraction of tumor cells accumulated β-catenin. Molecular analysis revealed that β-catenin gene mutations are a peculiar feature of skin tumors with matrical differentiation and correlate with a pattern of intense and diffuse β-catenin nuclear expression. In contrast, adenomatous polyposis coli (APC) and AXIN2 mutations were not involved in skin tumorigenesis. Analysis of Wnt pathway revealed that TCF-1 and MITF-M were selectively induced in the tumor types harboring β-catenin mutations, indicating that a Wnt/β-catenin pathway involving TCF-1 and MITF-M is activated in these tumors. Interestingly, high expression levels of TCF-3 were found in basal cell carcinomas and spiroadenomas. TCF-3 is reported to act as a negative modulator of β-catenin degradation pathway. Thus, the moderate increase of β-catenin nuclear staining detected in these tumor types might, at least in part, be due to a TCF-3-dependent mechanism. Finally, we found that the presence of β-catenin mutations significantly correlated with loss of nuclear immunoreactivity for an antibody raised against the N terminus of β-catenin (αABC). Thus, a combined analysis with C terminus-β-catenin antibodies and αABC Ab may represent a powerful investigative approach for the detection of β-catenin structural alterations. more...
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- 2003
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32. Uterine inflammatory myofibroblastic tumor in a 10-year-old girl presenting as polypoid mass
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Lorenza Pecciarini, Paolo Scollo, Maurilio Ponzoni, Paolo Amico, Giuseppe Pelosi, Filippo Fraggetta, Massimo Ippolito, Claudio Doglioni, Fraggetta, F, Doglioni, C, Scollo, P, Pecciarini, L, Ippolito, M1, Amico, P, Pelosi, G, and Ponzoni, Maurilio more...
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Inflammation ,Cancer Research ,medicine.medical_specialty ,business.industry ,media_common.quotation_subject ,Dermatology ,Immunohistochemistry ,Diagnosis, Differential ,Neoplasms, Muscle Tissue ,Polyps ,Oncology ,Lymphatic Metastasis ,Uterine Neoplasms ,medicine ,Biomarkers, Tumor ,Humans ,Female ,Girl ,business ,Child ,Watchful Waiting ,media_common - Published
- 2015
33. Abstract 744: ctDNA use in molecular diagnostics
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Greta Grassini, Maria Giulia Cangi, Lorenza Pecciarini, Ilaria Francaviglia, Claudio Doglioni, Elena DalCin, Alessandra Bulotta, Gilda Magliacane, Monika Ducceschi, Chiara Lazzari, and Vanesa Gregorc
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Oncology ,Cancer Research ,medicine.medical_specialty ,business.industry ,medicine.disease_cause ,Molecular diagnostics ,medicine.disease ,Cell-free fetal DNA ,Internal medicine ,medicine ,Carcinoma ,Digital polymerase chain reaction ,KRAS ,business ,Lung cancer ,Genotyping ,Allele frequency - Abstract
The analysis of circulating cell-free tumor DNA (ctDNA), which can be obtained from plasma by non-invasive procedures, is expected to provide useful biomarkers in the management of non-small-cell lung cancer (NSCLC) patients. Indeed, several studies have assessed ctDNA prognostic and predictive value as source of key data for therapeutic targets and drug resistance in carcinoma patients. The expanding number of targeted therapeutics for NSCLC always expects a real-time wider tumor genotyping, and the use of ctDNA as either a complement or an alternative to tumor tissue DNA (ttDNA) could be a valid option for Next-Generation Sequencing (NGS) of key cancer genes. However, isolation and enrichment of ctDNA is a big challenge because of its high degree of fragmentation and its low concentration against the normally occurring background of cell-free DNA derived from healthy cells. Therefore standardized methods for ctDNA extraction and analysis are crucial aspects in the setting-up of a molecular diagnostic approach. In this study we aimed to evaluate the ctDNA use for molecular profiling, also analyzing the impact of both pre-analytical and analytical variables on DNA yield and mutation detection. Matched ttDNA and ctDNA from 30 NSCLC patients were extracted, quantified and quality-controlled, and then investigated by different standard methods (real-time PCR, digital PCR, Mass Spectrometry genotyping) for EGFR, KRAS, BRAF, PIK3CA status. Mutational screening of ctDNA samples by IonTorrent NGS (Oncomine™ Lung cfDNA Assay, ThermoFisher) was also performed. Further, we extended ctDNA evaluation to additional 30 lung cancer patients with no available tumor sample. We found that cell free DNA concentration in plasma correlated with both stage and number of metastatic sites. Analyzing matched ttDNA and ctDNA by standard methods, we identified mutations in EGFR, KRAS, PIK3CA genes, with an overall concordance of 77%. Interestingly, Oncomine Lung cfDNA assay detected these same mutations with the same allelic frequency of standard methods; mutations in TP53 and ALK genes were also found. In the 30 patients with no available tumor sample, we found EGFR (10%) and KRAS (7%) mutations on ctDNA by standard methods; NGS analysis is under evaluation. This study evaluated the use of multiple different methods to detect mutations in NSCLC and showed that ctDNA can be a feasible option for clinical monitoring of lung cancers, including for those patients who cannot undergo invasive diagnostic procedures, due to either comorbidities or absence of biopsable tumor lesions. Citation Format: Gilda Magliacane, Greta Grassini, Ilaria Francaviglia, Elena DalCin, Chiara Lazzari, Alessandra Bulotta, Monika Ducceschi, Vanesa Gregorc, Lorenza Pecciarini, Claudio Doglioni, Maria Giulia Cangi. ctDNA use in molecular diagnostics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 744. doi:10.1158/1538-7445.AM2017-744 more...
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- 2017
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34. Abstract 3519: TGF-β pathway alteration in pancreatic neuroendocrine tumors: characterization of a novel SMAD3 translocation
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Luca Albarello, Maria Giulia Cangi, Michaela Bowden, Lorenza Pecciarini, Massimo Loda, Massimo Falconi, Emanuela Brunetto, Greta Grassini, Valeria De Pascali, Claudio Doglioni, Francesca Invernizzi, and Anna Talarico more...
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Cancer Research ,Oncology ,Chemistry ,Tgf β pathway ,medicine ,Cancer research ,Chromosomal translocation ,Neuroendocrine tumors ,medicine.disease - Abstract
Pancreatic neuroendocrine tumors (PanNETs) are the second most common malignancy of the pancreas, and the only curative option is represented by surgery. PanNETs can be classified as well- or poorly- differentiated, based on histology. Poorly-differentiated neuroendocrine carcinomas (NECs) represent less than 10% of cases, but they are highly malignant and associated with poor prognosis, and, most importantly, the inconsistency in their clinical management is due to the insufficient data on NEC genetic features. In order to widen our knowledge of PanNET genetic alterations, we performed RNASeq analysis (using Illumina NextSeq platform) of a group of 18 PanNETs and 2 NECs. TopHat-Fusion algorithm identified, among others, a fusion involving SMAD3 transcription factor and CCDC149; the alteration of SMAD3 in a poorly differentiated NEC was particularly interesting since SMAD3 is involved in TGF-b signaling, which has an oncogenic role in malignant cells. The SMAD3-CCDC149 fusion was confirmed in the index case by RT-PCR using primers spanning the breakpoint, followed by Sanger sequencing. The fusion was also detected by FISH analysis performed on both FFPE and frozen samples. By both FISH analysis and Sanger sequencing we identified two other poorly-differentiated NEC samples characterized by the presence of the SMAD3-CCDC149 translocation. Immunohistochemical (IHC) staining revealed that the 3 tumors, positive for the SMAD3-CCDC149 fusion, were characterized by nuclear localization of SMAD3. In order to test the hypothesis that the fusion generates an active form of SMAD3 that localizes exclusively within the nucleus, we transfected BON-1 cell line with plasmids containing either SMAD3 full length or SMAD3 truncated form (the latter mimics the fusion product) showing that only the truncated form of SMAD3 had an exclusively nuclear localization. Interestingly, FISH analysis performed on 3 well-differentiated PanNET samples did not detected the presence of the SMAD3-CDC149 translocation and IHC analysis showed cytoplasmic localization of SMAD3. Moreover, IHC analysis performed on a cohort of 40 cases of non-pancreatic neuroendocrine tumors (NETs) revealed that only poor-differentiated tumors showed a significant percentage of samples with nuclear localization of SMAD3. These data support the hypothesis that the translocation is a typical phenomenon of poorly-differentiated tumors and suggests that it could be involved also in non-pancreatic NETs. In this study we identified a previous unknown SMAD3 translocation in poor-differentiated NECs, which generates an active SMAD3 form. Deepening the knowledge of the fusion product will shed light on PanNET genetic alterations, opening the way to new therapeutic strategies. Citation Format: Emanuela Brunetto, Greta Grassini, Valeria De Pascali, Anna Talarico, Francesca Invernizzi, Michaela Bowden, Massimo Loda, Luca Albarello, Massimo Falconi, Lorenza Pecciarini, Maria G. Cangi, Claudio Doglioni. TGF-β pathway alteration in pancreatic neuroendocrine tumors: characterization of a novel SMAD3 translocation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3519. doi:10.1158/1538-7445.AM2017-3519 more...
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- 2017
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35. Abstract 756: Molecular characterization of triploid and tetraploid urine FISH samples
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Chiara Iacona, Massimo Freschi, Maria Giulia Cangi, Lorenza Pecciarini, Ilaria Francaviglia, Anna Talarico, Valeria De Pascali, and Claudio Doglioni
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Genetics ,Cancer Research ,Polysomy ,medicine.diagnostic_test ,Aneuploidy ,Biology ,medicine.disease ,medicine.disease_cause ,Molecular biology ,Molecular cytogenetics ,Oncology ,Chromosome 3 ,Tetrasomy ,medicine ,Trisomy ,Carcinogenesis ,Fluorescence in situ hybridization - Abstract
Urothelial carcinomas (UC) are characterized, among other aberrations, by chromosome 3, 7, 17 gains and 9p21 (p16) loss. Since UC cells readily exfoliate into the urine, molecular cytogenetics techniques have been used to detect cells consistent with UC diagnosis and follow-up: a multitarget Fluorescence In Situ Hybridization (FISH) test, composed of 3, 7 and 17 centromeres and 9p21 locus multicolor probes, has been proved useful for atypical cells identification in both void urine and lower and upper urinary tract samples before morphological changes are apparent through cytological and histological examination. To date, there are no uniform criteria for urine FISH scoring though the test is commonly considered positive following the UroVysion Bladder Cancer Kit criteria (Abbott Molecular): 4 or more cells with a gain of 2 or more chromosomes (polysomy), and/or 12 or more cells with 9p21 homozygous deletion. This definition of polysomy includes trisomy and tetrasomy classes, but, as tetraploidy may represent cells at S/G2 phase, this criterion may lead to false-positives; moreover it has been described that also inflammation induces tetraploidy, and that small percentages of normal urine cells show trisomy/tetrasomy. On the other hand, triploidy and tetraploidy may have a direct role in tumorigenesis, as an intermediate state in the development of cancer aneuploidy. At the moment little is reported about the clinical correlations of urine FISH cases showing only trisomy/tetrasomy and more data are needed to clarify the diagnostic value of triploid and tetraploid urothelial cells. Aiming to better classify patients with a trisomy/tetrasomy urine FISH result, we extracted genomic DNA from 50 fixed urine cell suspensions, previously tested by urine FISH, and investigated them for the presence of fibroblast growth factor receptor 3 (FGFR3) and telomerase reverse transcriptase (TERT) promoter mutations, which constitute the most recurrent somatic alterations in BC. In particular frequent mutations in exons 7, 10 and 15 of FGFR3 characterize non-muscle invasive UC while mutations in the core promoter region of TERT gene, mainly at -124 and -146 positions from the ATG start site, have emerged as the most frequent somatic genetic alteration in both non-muscle invasive and muscle invasive UC. All specimens were subjected to PCR amplification and Sanger sequencing for FGFR3 exon 7 and 10, and for the C228T and C250T TERT promoter mutations. A preliminary analysis of 20 urine FISH cases showed 4 samples with trisomy/tetrasomy, and 1 of these 4 had a mutation of the TERT promotor. Complete results of the FGFR3 and TERT promoter molecular analysis will be presented and correlated to both FISH results and patients clinical data, in order to evaluate if the presence of FGFR3 and/or TERT promoter mutations in urine samples with FISH detected trisomy/tetrasomy may help to identify patients with a higher risk of recurrence and progression. Citation Format: Lorenza Pecciarini, Valeria De Pascali, Ilaria Francaviglia, Anna Talarico, Chiara Iacona, Massimo Freschi, Maria Giulia Cangi, Claudio Doglioni. Molecular characterization of triploid and tetraploid urine FISH samples [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 756. doi:10.1158/1538-7445.AM2017-756 more...
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- 2017
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36. Identifying the primary sites of metastatic carcinoma: the increasing role of immunohistochemistry
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M. Giulia Cangi, Lorenza Pecciarini, and Claudio Doglioni
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Pathology ,medicine.medical_specialty ,Cytokeratin ,Primary sites ,Thyroid Transcription Factor 1 ,Unknown primary ,medicine ,Immunohistochemistry ,Biology ,Pathology and Forensic Medicine ,Site of origin ,Metastatic carcinoma ,Metastatic tumours - Abstract
Metastatic carcinoma from a tumour of unknown origin is a frequent clinical problem and identification of the primary site of origin may represent a difficult challenge for the clinician. Although most patients with metastatic carcinoma of unknown primary site have an ominous prognosis, there are subgroups of patients who are eligible for specific therapy and it is therefore mandatory to exploit every effort in order to identify accurately the tumour origin. The pathologist has acquired an increasing role in the characterization of the site of origin of these tumours by utilizing immunohistochemistry and molecular biological techniques. Recently, new powerful markers like cytokeratin 7 and 20, thyroid transcription factor 1 and Hep-1 have become available for the identification of the primary site of metastatic tumours, widening our limited repertoire of site-specific tumour antibodies. With the appropriate utilization of a restricted panel of antibodies it is now frequently possible to determine the site of origin of a metastatic carcinoma, solving several difficult clinico-pathological problems. However, this is not the end of the story: we still need more site- and tumour-specific markers in order to increase our diagnostic accuracy further. more...
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- 2001
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37. PAX8-PPAR γ 1 Fusion in Oncogene Human Thyroid Carcinoma
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Todd G. Kroll, Chang-Jie Chen, Elisabetta Mueller, Bruce M. Spiegelman, Lorenza Pecciarini, Pasha Sarraf, and Jonathan A. Fletcher
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chemistry.chemical_classification ,endocrine system ,Multidisciplinary ,endocrine system diseases ,Oncogene ,Thyroid ,Peroxisome proliferator-activated receptor ,Biology ,medicine.disease_cause ,medicine.disease ,Thyroid carcinoma ,Transactivation ,medicine.anatomical_structure ,chemistry ,medicine ,Cancer research ,Carcinoma ,Carcinogenesis ,PAX8 - Abstract
Chromosomal translocations that encode fusion oncoproteins have been observed consistently in leukemias/lymphomas and sarcomas but not in carcinomas, the most common human cancers. Here, we report that t(2;3)(q13;p25), a translocation identified in a subset of human thyroid follicular carcinomas, results in fusion of the DNA binding domains of the thyroid transcription factor PAX8 to domains A to F of the peroxisome proliferator–activated receptor (PPAR) γ1. PAX8-PPARγ1 mRNA and protein were detected in 5 of 8 thyroid follicular carcinomas but not in 20 follicular adenomas, 10 papillary carcinomas, or 10 multinodular hyperplasias. PAX8-PPARγ1 inhibited thiazolidinedione-induced transactivation by PPARγ1 in a dominant negative manner. The experiments demonstrate an oncogenic role for PPARγ and suggest that PAX8-PPARγ1 may be useful in the diagnosis and treatment of thyroid carcinoma. more...
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- 2000
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38. A 'twist box' code of p53 inactivation: twist box: p53 interaction promotes p53 degradation
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Lorenza Pecciarini, Sara Piccinin, Sabrina Rossi, Camillo Rosano, Flavia Pivetta, Angelo Paolo Dei Tos, Sara Sessa, Claudio Doglioni, Alessandra Grizzo, Elena Tonin, Lucia Zanatta, Maura Sonego, Roberta Maestro, Silvia Demontis, Piccinin, S, Tonin, E, Sessa, S, Demontis, S, Rossi, S, Pecciarini, L, Zanatta, L, Pivetta, F, Grizzo, A, Sonego, M, Rosano, C, Dei Tos, Ap, Doglioni, Claudio, and Maestro, R. more...
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Cancer Research ,Epithelial-Mesenchymal Transition ,animal structures ,DNA Copy Number Variations ,Mice, Nude ,Biology ,Tp53 mutation ,Mice ,medicine ,Animals ,Humans ,Phosphorylation ,RNA, Small Interfering ,Twist ,Genetics ,C-terminus ,Twist-Related Protein 1 ,Nuclear Proteins ,Proto-Oncogene Proteins c-mdm2 ,Sarcoma ,Cell Biology ,Cancer treatment ,Cell biology ,Repressor Proteins ,Cell Transformation, Neoplastic ,Mechanism of action ,Oncology ,biology.protein ,Mdm2 ,RNA Interference ,Cancer development ,Tumor Suppressor Protein p53 ,medicine.symptom - Abstract
Twist proteins have been shown to contribute to cancer development and progression by impinging on different regulatory pathways, but their mechanism of action is poorly defined. By investigating the role of Twist in sarcomas, we found that Twist1 acts as a mechanism alternative to TP53 mutation and MDM2 overexpression to inactivate p53 in mesenchymal tumors. We provide evidence that Twist1 binds p53 C terminus through the Twist box. This interaction hinders key posttranslational modifications of p53 and facilitates its MDM2-mediated degradation. Our study suggests the existence of a Twist box code of p53 inactivation and provides the proof of principle that targeting the Twist box:p53 interaction might offer additional avenues for cancer treatment. Twist proteins have been shown to contribute to cancer development and progression by impinging on different regulatory pathways, but their mechanism of action is poorly defined. By investigating the role of Twist in sarcomas, we found that Twist1 acts as a mechanism alternative to TP53 mutation and MDM2 overexpression to inactivate p53 in mesenchymal tumors. We provide evidence that Twist1 binds p53 C terminus through the Twist box. This interaction hinders key posttranslational modifications of p53 and facilitates its MDM2-mediated degradation. Our study suggests the existence of a Twist box code of p53 inactivation and provides the proof of principle that targeting the Twist box:p53 interaction might offer additional avenues for cancer treatment. more...
- Published
- 2012
39. HER2 Testing in Gastric Cancer
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Lorenza Pecciarini, Luca Albarello, Claudio Doglioni, Albarello, Luca, Pecciarini, Lorenza, and Doglioni, Claudio
- Subjects
Oncology ,medicine.medical_specialty ,Receptor, ErbB-2 ,medicine.medical_treatment ,Breast Neoplasms ,Antibodies, Monoclonal, Humanized ,Pathology and Forensic Medicine ,Trastuzumab ,Stomach Neoplasms ,Internal medicine ,Carcinoma ,medicine ,Humans ,skin and connective tissue diseases ,neoplasms ,In Situ Hybridization, Fluorescence ,Randomized Controlled Trials as Topic ,Chemotherapy ,business.industry ,Stomach ,Gene Amplification ,Cancer ,Antibodies, Monoclonal ,medicine.disease ,Immunohistochemistry ,Clinical trial ,medicine.anatomical_structure ,Clinical Trials, Phase III as Topic ,Monoclonal ,Esophagogastric Junction ,Anatomy ,Breast carcinoma ,business ,medicine.drug - Abstract
Molecular therapies targeting HER2 are part of the established drug armamentarium in breast carcinoma. Now the ToGA trial, an international multicenter phase III clinical study, involving 24 countries globally, has shown that the anti-HER2 humanized monoclonal antibody Trastuzumab is effective in prolonging survival in HER2-positive carcinoma of the stomach and the gastroesophageal junction (GEJ). Similarly to breast carcinoma, > 20% of gastric cancers show HER2 overexpression and/or amplification, and this percentage increases to 33% in GEJ tumors. Thus, as in breast carcinoma, pathologists are now asked to evaluate HER2 status in gastric carcinoma samples. As validated in the ToGA trial, the HER2 testing criteria that must be used in evaluating both gastric carcinoma biopsies and surgical specimens significantly differ from those routinely applied in breast carcinoma. The main variations with regard to the pattern of reactivity in HER2-expressing cells are as follows: the completeness of membrane staining is not a "conditio sine qua non" and the number of stained cells necessary to consider a case as positive is different. We must also take note of the much more frequent heterogeneity of HER2 positivity in gastric cancer compared with breast carcinoma and the less stringent correlation between HER2 amplification and protein overexpression that is observed in gastric carcinoma, where more than 20% of cases may carry HER2 amplification, although of low level, without HER2 expression. In these patients, in the ToGA trial, there was no apparent benefit from adding Trastuzumab to chemotherapy: for this reason the European Medicines Agency, while approving usage of Trastuzumab for metastatic adenocarcinoma treatment, indicated HER2 testing by immunohistochemistry as first evaluation assay, followed by fluorescence in situ hybridization in 2+ equivocal cases. HER2 testing in gastric carcinoma is a new field, opening several opportunities: for patients with gastric cancer, this is a new promising therapeutic option; for pathologists, strengthening our role in therapy selection and emphasizing our duty of providing accurate and reproducible HER2 testing results; for all interested in understanding the biology of gastric and GEJ cancer and in discovering new possible molecular therapy targets. ZR 0 ZS 0 ZB 51 Z8 7 more...
- Published
- 2011
40. Urocortin in amniotic fluid and Down syndrome
- Author
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Michela, Torricelli, Chiara, Voltolini, Giulia, Biliotti, Claudio, Giorlandino, Flavio, De Pascalis, Maria, De Bonis, Alvaro, Mesuraca, Alessia, Giovannelli, Lorenza, Pecciarini, and Felice, Petraglia more...
- Subjects
Adult ,Pregnancy ,Case-Control Studies ,Pregnancy Trimester, Second ,Amniocentesis ,Humans ,Female ,Gestational Age ,Down Syndrome ,Amniotic Fluid ,Urocortins ,Retrospective Studies - Published
- 2009
41. Chlamydia infection and lymphomas: association beyond ocular adnexal lymphomas highlighted by multiple detection methods
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Elena Dal Cin, Maurilio Ponzoni, S. Magnino, Lorenza Pecciarini, Riccardo Dolcetti, Elisa Pasini, Stefano Grassi, Luciano Sacchi, Rosalba Stefano, Antonia A. Lettini, Massimo Guidoboni, Claudio Doglioni, Maria Giulia Cangi, Andrés J.M. Ferreri, Ponzoni, Maurilio, Ferreri, Ajm, Guidoboni, M, Lettini, Aa, Cangi, Mg, Pasini, E, Sacchi, L, Pecciarini, L, Grassi, S, Dal Cin, E, Stefano, R, Magnino, S, Dolcetti, R, and Doglioni, Claudio more...
- Subjects
DNA, Bacterial ,Cancer Research ,Pathology ,medicine.medical_specialty ,Lymphoma ,Immunofluorescence ,Eye neoplasm ,Polymerase Chain Reaction ,Monocytes ,medicine ,Humans ,Microdissection ,Chlamydia psittaci ,Chlamydia ,medicine.diagnostic_test ,biology ,Eye Neoplasms ,Macrophages ,MALT lymphoma ,Chlamydia Infections ,Psittacosis ,medicine.disease ,biology.organism_classification ,Oncology ,Chlamydophila psittaci ,Immunology ,Immunohistochemistry - Abstract
Purpose: Chlamydia psittaci (Cp) has been associated to ocular adnexal lymphomas (OAL) with variable geographic distribution. Herein, we used multiple Chlamdia detection tools to identify Cp elementary bodies-containing cell and to assess Cp prevalence in both nodal and extranodal lymphomas. Experimental Design: TETR-PCR, immunohistochemistry, immunofluorescence, electron microscopy, and laser-capture microdissection were done in 35 OALs to define their effect in Chlamydia detection and, moreover, to identify the Cp cellular carrier. Cp prevalence was screened by TETR-PCR in 205 extraorbital lymphomas and 135 nonneoplastic controls. Results: Twenty-six (74%) OALs were associated with Cp infection: immunohistochemistry, immunofluorescence, and laser-capture microdissection-assisted PCR showed that monocytes/macrophages were the Cp carriers; electron microscopy showed the presence of intact CP elementary bodies into these cells. Immunohistochemistry and TETR-PCR showed a 70% concordance rate (P = 0.001). Cp DNA was equally prevalent in non-OAL, nodal, and extranodal lymphomas: among the latter, it was more common in diffuse large B-cell lymphomas of the skin (P = 0.03) and Waldeyer's ring. Conclusions: This multiparametric approach shows, for the first time, that monocytes/macrophages are the carriers of Cp, Cp seems preferentially associated with lymphomas arising in organs primarily exposed to antigens. The clinical implications of these findings deserve to be prospectively investigated. Purpose: Chlamydia psittaci (Cp) has been associated to ocular adnexal lymphomas (OAL) with variable geographic distribution. Herein, we used multiple Chlamdia detection tools to identify Cp elementary bodies-containing cell and to assess Cp prevalence in both nodal and extranodal lymphomas. Experimental Design: TETR-PCR, immunohistochemistry, immunofluorescence, electron microscopy, and laser-capture microdissection were done in 35 OALs to define their effect in Chlamydia detection and, moreover, to identify the Cp cellular carrier. Cp prevalence was screened by TETR-PCR in 205 extraorbital lymphomas and 135 nonneoplastic controls. Results: Twenty-six (74%) OALs were associated with Cp infection: immunohistochemistry, immunofluorescence, and laser-capture microdissection-assisted PCR showed that monocytes/macrophages were the Cp carriers; electron microscopy showed the presence of intact CP elementary bodies into these cells. Immunohistochemistry and TETR-PCR showed a 70% concordance rate (P = 0.001). Cp DNA was equally prevalent in non-OAL, nodal, and extranodal lymphomas: among the latter, it was more common in diffuse large B-cell lymphomas of the skin (P = 0.03) and Waldeyer's ring. Conclusions: This multiparametric approach shows, for the first time, that monocytes/macrophages are the carriers of Cp, Cp seems preferentially associated with lymphomas arising in organs primarily exposed to antigens. The clinical implications of these findings deserve to be prospectively investigated. PURPOSE: Chlamydia psittaci (Cp) has been associated to ocular adnexal lymphomas (OAL) with variable geographic distribution. Herein, we used multiple Chlamydia detection tools to identify Cp elementary bodies-containing cell and to assess Cp prevalence in both nodal and extranodal lymphomas. EXPERIMENTAL DESIGN: TETR-PCR, immunohistochemistry, immunofluorescence, electron microscopy, and laser-capture microdissection were done in 35 OALs to define their effect in Chlamydia detection and, moreover, to identify the Cp cellular carrier. Cp prevalence was screened by TETR-PCR in 205 extraorbital lymphomas and 135 nonneoplastic controls. RESULTS: Twenty-six (74%) OALs were associated with Cp infection: immunohistochemistry, immunofluorescence, and laser-capture microdissection-assisted PCR showed that monocytes/macrophages were the Cp carriers; electron microscopy showed the presence of intact Cp elementary bodies into these cells. Immunohistochemistry and TETR-PCR showed a 70% concordance rate (P = 0.001). Cp DNA was equally prevalent in non-OAL, nodal, and extranodal lymphomas: among the latter, it was more common in diffuse large B-cell lymphomas of the skin (P = 0.03) and Waldeyer's ring. CONCLUSIONS: This multiparametric approach shows, for the first time, that monocytes/macrophages are the carriers of Cp, Cp seems preferentially associated with lymphomas arising in organs primarily exposed to antigens. The clinical implications of these findings deserve to be prospectively investigated. more...
- Published
- 2008
42. Constitutive overexpression of CDC25A in primary human mammary epithelial cells results in both defective DNA damage response and chromosomal breaks at fragile sites
- Author
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Sara Piccinin, Anna Talarico, Lorenza Pecciarini, Stefano Grassi, Claudio Doglioni, Elena Dal Cin, Roberta Maestro, Alessandra Grizzo, Maria Giulia Cangi, Cangi, Mg, Piccinin, S, Pecciarini, L, Talarico, A, DAL CIN, E, Grassi, S, Grizzo, A, Maestro, R, and Doglioni, Claudio more...
- Subjects
Aphidicolin ,Genome instability ,Cancer Research ,CDC25A ,Cell cycle checkpoint ,DNA Repair ,DNA damage ,Blotting, Western ,Fluorescent Antibody Technique ,Breast Neoplasms ,Biology ,chemistry.chemical_compound ,Humans ,cdc25 Phosphatases ,Mammary Glands, Human ,Cells, Cultured ,In Situ Hybridization, Fluorescence ,Chromosomal fragile site ,Cell Cycle ,DNA replication ,Epithelial Cells ,Cell cycle ,Up-Regulation ,Cell Transformation, Neoplastic ,Oncology ,chemistry ,Cancer research ,Female ,DNA Damage - Abstract
CDC25A phosphatase, an essential component of the cell cycle machinery, is also a key player in integrating the specific signals of checkpoint control in response to DNA damage. There are several lines of evidence that indicate a role for CDC25A in cancer (level opment, consistent with the fact that its overexpression is detected in human cancers. In particular we previously reported that CDC25A is overexpressed also in early breast carcinoma. Recent data suggest that oncogene activation during early stages of tumor development causes DNA replication stress resulting in the induction of DNA damage response (DDR) and that the selection of cells defecting in their DDR could lead to malignant progression. To address how CDC25A overexpression contributes to breast cancer development we established it cell model in which CDC25A was constitutively overexpressed in hTERT-immortalized primary human mammary epithelial cells. At the earliest passages following CDC25A transduction we observed DDR signs associated with unscheduled DNA replication origins. In the latest passages DDR was significantly impaired and, even after ionizing radiation exposition, cells failed to induce G1 and G2 checkpoints; moreover DNA replication stress conditions, such as aphidicolin treatment, highlighted increased fragile site breakages and destabilized chromosomes Just in these latest passages cells. Our data suggest that CDC25A overexpression, pushing the cell through the cell cycle transitions, induces DDR alterations that might enhance genomic instability. (C) 2008 Wiley-Liss, Inc. OI Maestro, Roberta/0000-0002-6642-5592 ZR 0 ZS 0 Z8 1 CDC25A phosphatase, an essential component of the cell cycle machinery, is also a key player in integrating the specific signals of checkpoint control in response to DNA damage. There are several lines of evidence that indicate a role for CDC25A in cancer development, consistent with the fact that its overexpression is detected in human cancers. In particular we previously reported that CDC25A is overexpressed also in early breast carcinoma. Recent data suggest that oncogene activation during early stages of tumor development causes DNA replication stress resulting in the induction of DNA damage response (DDR) and that the selection of cells defecting in their DDR could lead to malignant progression. To address how CDC25A overexpression contributes to breast cancer development we established a cell model in which CDC25A was constitutively overexpressed in hTERT-immortalized primary human mammary epithelial cells. At the earliest passages following CDC25A transduction we observed DDR signs associated with unscheduled DNA replication origins. In the latest passages DDR was significantly impaired and, even after ionizing radiation exposition, cells failed to induce G1 and G2 checkpoints; moreover DNA replication stress conditions, such as aphidicolin treatment, highlighted increased fragile site breakages and destabilized chromosomes just in these latest passages cells. Our data suggest that CDC25A overexpression, pushing the cell through the cell cycle transitions, induces DDR alterations that might enhance genomic instability. more...
- Published
- 2008
43. Chlamydia-psittaci-eradicating antibiotic therapy in patients with advanced-stage ocular adnexal MALT lymphoma
- Author
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G.P. Dognini, G. Santambrogio, M. Ponzoni, Antonio Giordano Resti, Andrés J.M. Ferreri, Nadia Vicari, S. Magnino, Claudio Doglioni, Maria Giulia Cangi, Elisa Pasini, C. De Conciliis, Lorenza Pecciarini, Riccardo Dolcetti, Ferreri, Ajm, Dognini, Gp, Ponzoni, Maurilio, Pecciarini, L, Cangi, Mg, Santambrogi, G, Resti, Ag, De Conciliis, C, Magnino, S, Pasini, E, Vicari, N, Dolcetti, R, and Doglioni, Claudio more...
- Subjects
Male ,Pathology ,medicine.medical_specialty ,Psittacosis ,Subcutaneous Tissue ,Ocular Adnexal MALT lymphoma ,Antibiotic therapy ,medicine ,Humans ,In patient ,Aged ,Doxycycline ,Chlamydia psittaci ,Aged, 80 and over ,biology ,business.industry ,Advanced stage ,Remission Induction ,Hematology ,Lymphoma, B-Cell, Marginal Zone ,Middle Aged ,medicine.disease ,biology.organism_classification ,Dermatology ,Lymphoma ,Anti-Bacterial Agents ,Oncology ,Chlamydophila psittaci ,Orbital Neoplasms ,Female ,business ,medicine.drug - Published
- 2007
44. A transcription-dependent micrococcal nuclease-resistant fragment of the urokinase-type plasminogen activator promoter interacts with the enhancer
- Author
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Francesco Blasi, Paolo Luraghi, Lorenza Pecciarini, Davide Munari, Carmelo Ferrai, Maria Giulia Cangi, Claudio Doglioni, Massimo P. Crippa, Ferrai, C, Munari, D, Luraghi, P, Pecciarini, L, Cangi, Mg, Doglioni, Claudio, Blasi, F, and Crippa, Mp more...
- Subjects
Chromatin Immunoprecipitation ,Models, Genetic ,Transcription, Genetic ,biology ,Cell Biology ,Urokinase-Type Plasminogen Activator ,Biochemistry ,Molecular biology ,Chromatin ,Rats ,Enhancer Elements, Genetic ,Transcription (biology) ,biology.protein ,Transcriptional regulation ,Animals ,Micrococcal Nuclease ,Promoter Regions, Genetic ,Enhancer ,Molecular Biology ,Chromatin immunoprecipitation ,Plasminogen activator ,Gene ,Micrococcal nuclease - Abstract
We show the interaction between the enhancer and the minimal promoter of urokinase-type plasminogen activator gene during active transcription by coupling micrococcal nuclease digestion of cross-linked, sonicated chromatin, and chromatin immunoprecipitation. This approach allowed the precise identification of the interacting genomic fragments, one of which is resistant to micrococcal nuclease cleavage. The interacting fragments form a single transcriptional control unit, as indicated by their common protein content. Furthermore, we show that the enhancer-MP interaction persists during the early stages of transcription and is lost upon alpha-amanitin treatment, indicating the requirement for active transcription. Our results support a looping model of interaction between the enhancer and the MP of the urokinase-type plasminogen activator gene. more...
- Published
- 2007
45. A woman and her canary: a tale of chlamydiae and lymphomas
- Author
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Lorenza Pecciarini, Riccardo Dolcetti, S. Magnino, G.P. Dognini, Antonio Giordano Resti, Maurilio Ponzoni, Claudio Doglioni, Antonis Dagklis, Elisa Pasini, Nadia Vicari, Paolo Ghia, Andrés J.M. Ferreri, Maria Giulia Cangi, Dolcetti, R, Magnino, S, Doglioni, Claudio, Cangi, Mg, Pecciarini, L, Ghia, PAOLO PROSPERO, Dagklis, A, Pasini, E, Vicari, N, Dognini, Gp, Resti, Ag, and Ponzoni, Maurilio more...
- Subjects
Cancer Research ,Oncology ,biology ,Chlamydiales ,Immunology ,Chlamydophila infections ,medicine ,Chlamydiae ,Second primary cancer ,medicine.disease ,biology.organism_classification ,Virology ,Lymphoma - Published
- 2007
46. Alterations of beta-catenin pathway in non-melanoma skin tumors: loss of alpha-ABC nuclear reactivity correlates with the presence of beta-catenin gene mutation
- Author
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Claudio, Doglioni, Sara, Piccinin, Silvia, Demontis, Maria Giulia, Cangi, Lorenza, Pecciarini, Concetta, Chiarelli, Michela, Armellin, Tamara, Vukosavljevic, Mauro, Boiocchi, and Roberta, Maestro
- Subjects
Cell Nucleus ,Genes, APC ,Skin Neoplasms ,DNA Mutational Analysis ,Antibodies, Monoclonal ,Immunohistochemistry ,Cytoskeletal Proteins ,Axin Protein ,Mutation ,Trans-Activators ,Humans ,beta Catenin ,Skin ,Regular Articles - Abstract
To determine the role of beta-catenin pathway in human skin carcinogenesis, 135 non-melanoma skin tumors were analyzed for beta-catenin expression and gene mutations. Intense nucleo-cytoplasmic immunoreactivity for C terminus beta-catenin antibodies was observed in all pilomatricomas and in single cases of trichoepithelioma and squamous cell carcinoma showing peculiar signs of matrical differentiation. Moderate increase of beta-catenin nuclear staining was detected in a significant proportion of basal cell carcinomas, Bowen disease, spiroadenomas, and occasionally also in squamous cell carcinomas, but in these neoplasms only a limited fraction of tumor cells accumulated beta-catenin. Molecular analysis revealed that beta-catenin gene mutations are a peculiar feature of skin tumors with matrical differentiation and correlate with a pattern of intense and diffuse beta-catenin nuclear expression. In contrast, adenomatous polyposis coli (APC) and AXIN2 mutations were not involved in skin tumorigenesis. Analysis of Wnt pathway revealed that TCF-1 and MITF-M were selectively induced in the tumor types harboring beta-catenin mutations, indicating that a Wnt/beta-catenin pathway involving TCF-1 and MITF-M is activated in these tumors. Interestingly, high expression levels of TCF-3 were found in basal cell carcinomas and spiroadenomas. TCF-3 is reported to act as a negative modulator of beta-catenin degradation pathway. Thus, the moderate increase of beta-catenin nuclear staining detected in these tumor types might, at least in part, be due to a TCF-3-dependent mechanism. Finally, we found that the presence of beta-catenin mutations significantly correlated with loss of nuclear immunoreactivity for an antibody raised against the N terminus of beta-catenin (alphaABC). Thus, a combined analysis with C terminus-beta-catenin antibodies and alphaABC Ab may represent a powerful investigative approach for the detection of beta-catenin structural alterations. more...
- Published
- 2003
47. High syndecan-1 expression in breast carcinoma is related to an aggressive phenotype and to poorer prognosis
- Author
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Enzo Galligioni, Claudio Doglioni, Francesco Mauri, Mattia Barbareschi, A. Lucenti, Orazio Caffo, Silvio Veronese, Patrick Maisonneuve, Maria Giulia Cangi, Lorenza Pecciarini, D. Aldovini, Paolo Dalla Palma, Barbareschi, M, Maisonneuve, P, Aldovini, D, Cangi, Mg, Pecciarini, L, Mauri, Fa, Veronese, S, Caffo, O, Lucenti, A, Palma, Pd, Galligioni, E, and Doglioni, Claudio more...
- Subjects
Oncology ,Cancer Research ,medicine.medical_specialty ,animal structures ,Syndecans ,Receptor, ErbB-2 ,Estrogen receptor ,Antineoplastic Agents ,Breast Neoplasms ,Adenocarcinoma ,Syndecan 1 ,Immunoenzyme Techniques ,Internal medicine ,Adjuvant therapy ,Carcinoma ,Medicine ,Humans ,RNA, Messenger ,RNA, Neoplasm ,Survival rate ,Survival analysis ,DNA Primers ,Membrane Glycoproteins ,business.industry ,Reverse Transcriptase Polymerase Chain Reaction ,Cancer ,Middle Aged ,medicine.disease ,Prognosis ,carbohydrates (lipids) ,Survival Rate ,Endocrinology ,Receptors, Estrogen ,Chemotherapy, Adjuvant ,Lymphatic Metastasis ,Female ,Proteoglycans ,Syndecan-1 ,Stromal Cells ,business ,Breast carcinoma ,Receptors, Progesterone ,Follow-Up Studies - Abstract
BACKGROUND Syndecan-1 is a transmembrane heparan sulphate proteoglycan that is involved in cell–cell adhesion, organization of cell–matrix adhesion, and regulation of growth factor signaling. METHODS Specimens from 254 consecutive breast carcinoma (BC) cases (110 N0, 144 N1/2) with long-term follow-up (median, 95 months) were immunostained for syndecan-1, estrogen receptor (ER), progesterone receptor (PgR), and p53; in 154 cases, c-erbB-2 status was known. Syndecan-1 mRNA and protein expression also were evaluated in 20 breast tissue samples (10 normal and tumor pairs). RESULTS Syndecan-1 was expressed at high levels in 106 (42%) BCs; syndecan-1 up-regulation was confirmed by reverse transcriptase–polymerase chain reaction (RT-PCR) studies. High syndecan-1 expression was associated with high histologic grade, large tumor size, high mitotic count, c-erbB-2 overexpression, and ER and PgR negative status. At univariate survival analysis syndecan overexpression was related to poor prognosis (P < 0.01 for both overall survival (OS) and disease-free survival). Bivariate survival analysis showed an additive adverse effect for syndecan-1 and c-erbB-2 overexpression. At multivariate analysis, syndecan-1 overexpression was independently associated with poor OS (hazard ratio [HR], 1.71; 95% confidence interval [CI], 1.08–2.69). High syndecan-1 expression also was of independent prognostic value for OS in the group of 102 ER-negative patients (HR, 2.42; 95% CI, 1.21–4.82). Stratifying patients on the basis of the type of adjuvant therapy given, high syndecan-1 expression was associated with a higher risk of death only in patients treated with the cyclophosphamide-methotrexate-fluorouracil regimen (HR, 1.9; P = 0.09); at multivariate analysis for OS, this association proved to be of independent statistical significance (P = 0.03; HR, 2.15). CONCLUSIONS Syndecan-1 is expressed at high levels in a significant percentage of breast carcinomas and is related to an aggressive phenotype and poor clinical behavior. Cancer 2003;98:474–83. © 2003 American Cancer Society. DOI 10.1002/cncr.11515 more...
- Published
- 2003
48. p63, a p53 homologue, is a selective nuclear marker of myoepithelial cells of the human breast
- Author
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Claudio Doglioni, Ettore Macrì, Lorenza Pecciarini, Giuseppe Viale, Maria Giulia Cangi, Aroldo Rizzo, Mattia Barbareschi, Barbareschi, M, Pecciarini, L, Cangi, Mg, Macri, E, Rizzo, A, Viale, G, and Doglioni, Claudio more...
- Subjects
Genetic Markers ,Pathology ,medicine.medical_specialty ,Stromal cell ,Breast Neoplasms ,Biology ,Pathology and Forensic Medicine ,Biomarkers, Tumor ,medicine ,Humans ,Genes, Tumor Suppressor ,Breast ,RNA, Messenger ,Progenitor cell ,Lymph node ,DNA Primers ,Cell Nucleus ,Reverse Transcriptase Polymerase Chain Reaction ,Tumor Suppressor Proteins ,Carcinoma ,Myoepithelial cell ,Membrane Proteins ,Epithelial Cells ,Ductal carcinoma ,Genes, p53 ,Phosphoproteins ,medicine.disease ,Immunohistochemistry ,Squamous metaplasia ,DNA-Binding Proteins ,medicine.anatomical_structure ,Trans-Activators ,Female ,Surgery ,sense organs ,Anatomy ,Stem cell ,Transcription Factors - Abstract
Myoepithelial cells (MCs) constitute the basal cell layer of normal mammary epithelia, and their identification is of particular diagnostic value because they are retained in most benign lesions while being lost in malignancy. Several MC immunocytochemical markers are currently available for diagnostic purposes, with special reference to smooth muscle-related antigens. p63 is a member of the p53 gene family, and its germline mutations are associated with severe mammary developmental defects in both rodents and humans. Different p63 isoforms have been identified, some of which (Delta Np63) are preferentially expressed in the epithelial basal cells of different or-ans and have been considered as possible markers of stem cells/reserve cells. We investigated immunohistochemically 384 samples of normal and diseased human breast, including 300 invasive carcinomas, using four antibodies recognizing all p63 isoforms, or the Delta Np63 isoforms. Twenty cytologic specimens were also investigated. Furthermore, snap-frozen tissue samples from three fibroadenomas and 10 invasive ductal carcinomas with their paired non-neoplastic tissues and three corresponding lymph node metastases were evaluated for the expression of p63 mRNA by RT-PCR. In normal breast tissue p63 immunoreactivity was confined to the nuclei of MCs. In all benign lesions p63-immunoreactive cells formed a continuous basal rim along the epithelial structures. Stromal cells, and in particular myofibroblasts, were consistently unreactive. Adenomyoepitheliomas showed nuclear staining in most neoplastic cells. A peripheral rim of p63-immunoreactive cells was retained surrounding lobular and ductal carcinoma in situ, although it was discontinuous as opposed to the normal structures. Invasive breast carcinomas were consistently devoid of nuclear p63 staining, with the exception of the two adenoid-cystic carcinomas, of the two ductal carcinomas with squamous metaplasia, and of 11 (4.6%) ductal carcinomas not otherwise specified, showing p63 immunoreactivity in a minor fraction (5-15%) of the neoplastic cells. In comparison with other MC markers, p63 was the most specific, being restricted exclusively to MCs, whereas antibodies to smooth muscle actin and, to a lesser extent, calponin also decorated stromal myofibroblasts. In the cytologic preparations p63 immunoreactivity was a consistent feature of "naked nuclei" and of a subset of cells surrounding benign epithelial clusters. RT-PCR experiments with primers specific for different p63 isoforms documented that normal tissues and fibroadenomas preferentially expressed the Delta Np63 isoforms. Our study demonstrates that in normal and pathologic breast tissues MCs consistently express the Delta Np63 isoforms. We suggest p63 as a reliable, highly specific, and sensitive MC marker in both histologic and cytologic preparations. Furthermore, because p63 immunoreactivity in adult epithelia is normally restricted to progenitor cells, it can be speculated that it mi-ht be a clue for the identification of the still elusive breast progenitor cells. ZR 0 Z8 16 ZB 189 ZS 2 more...
- Published
- 2001
49. Urocortin in amniotic fluid and Down syndrome
- Author
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Lorenza Pecciarini, Alessia Giovannelli, Giulia Biliotti, Felice Petraglia, Michela Torricelli, Claudio Giorlandino, Alvaro Mesuraca, Flavio De Pascalis, Chiara Voltolini, and Maria De Bonis
- Subjects
Urocortin ,Down syndrome ,medicine.medical_specialty ,Amniotic fluid ,medicine.diagnostic_test ,Obstetrics and Gynecology ,Biology ,medicine.disease ,Endocrinology ,Internal medicine ,medicine ,Amniocentesis ,Trisomy ,Genetics (clinical) - Published
- 2009
- Full Text
- View/download PDF
50. Small thymus in very low birth weight infants born to mothers with subclinical chorioamnionitis
- Author
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Franco Bagnoli, Lorenza Pecciarini, Paolo Toti, Claudio De Felice, Rosa Santopietro, and Michela Stumpo
- Subjects
Male ,medicine.medical_specialty ,thymus ,chorioamnionitis ,preterm ,Thymus Gland ,Chorioamnionitis ,Small thymus ,Atrophy ,Pregnancy ,Humans ,Infant, Very Low Birth Weight ,Medicine ,Risk factor ,Subclinical infection ,business.industry ,Obstetrics ,Infant, Newborn ,Case-control study ,medicine.disease ,Radiography ,Low birth weight ,Case-Control Studies ,Pediatrics, Perinatology and Child Health ,Female ,medicine.symptom ,business - Abstract
Chorioamnionitis, a major cause of preterm birth with significant neonatal morbidity and mortality, frequently occurs in mothers who are free of symptoms. A combined clinical, radiologic, and pathologic study of 129 very low birth weight infants indicated a significant association between a markedly decreased thymic size at birth and subclinical chorioamnionitis. more...
- Published
- 1999
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