67 results on '"Pecciarini L"'
Search Results
2. Prospective Validation of the Italian Alliance Against Cancer Lung Panel in Patients With Advanced Non–Small-Cell Lung Cancer
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Gregorc, V., Mazzarella, L., Lazzari, C., Graziano, P., Vigneri, P., Genova, C., Toschi, L., Ciliberto, G., Bonanno, L., Delmonte, A., Bucci, G., Rossi, A., Motta, G., Coco, S., Marinello, A., Buglioni, S., Cangi, M. G., Di Micco, C., Bandiera, A., Bonfiglio, S., Pecciarini, L., Guida, A., Ceol, A., Frige', G., De Maria Marchiano, Ruggero, Pelicci, P. G., De Maria R. (ORCID:0000-0003-2255-0583), Gregorc, V., Mazzarella, L., Lazzari, C., Graziano, P., Vigneri, P., Genova, C., Toschi, L., Ciliberto, G., Bonanno, L., Delmonte, A., Bucci, G., Rossi, A., Motta, G., Coco, S., Marinello, A., Buglioni, S., Cangi, M. G., Di Micco, C., Bandiera, A., Bonfiglio, S., Pecciarini, L., Guida, A., Ceol, A., Frige', G., De Maria Marchiano, Ruggero, Pelicci, P. G., and De Maria R. (ORCID:0000-0003-2255-0583)
- 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.
- Published
- 2021
3. Chlamydia psittaci-eradicating antibiotic therapy in patients with advanced-stage ocular adnexal MALT lymphoma
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Ferreri, A. J. M., Dognini, G. P., Ponzoni, M., Pecciarini, L., Cangi, M. G., Santambrogio, G., Resti, A. G., De Conciliis, C., Magnino, S., Pasini, E., Vicari, N., Dolcetti, R., and Doglioni, C.
- Published
- 2008
4. Modeling multiple myeloma-bone marrow interactions and response to drugs in a 3d surrogate microenvironment
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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, Ferrero, Elisabetta, 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
- 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 myelomabone marrow co-cultures in gelatin scaffolds. The model was validated with myeloma cell lines that, as assessed by histochemical and electronmicroscopic analyses, engaged contacts with stromal cells and endothelial cells. Consistently, pro-survival signaling and also cell adhesionmediated 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 highrisk 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.
- Published
- 2018
5. Human malignant mesothelioma is recapitulated in immunocompetent BALB/c mice injected with murine AB cells
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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, Crippa, M., Valtorta, Silvia, MORESCO, ROSA MARIA, 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, Crippa, M., Valtorta, Silvia, and MORESCO, ROSA MARIA
- 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.
- Published
- 2016
6. Bcl2, Bcl6, MYC, MALT 1 and Bcl10 rearrangements in nodal diffuse large B-cell lymphomas: A multicenter evaluation of a new set of fish probes and correlation with clinical outcome
- Author
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Tibiletti, M., Martin, V., Bernasconi, B., Del Curto, B., Pecciarini, L., Uccella, S., Giancarlo Pruneri, Ponzoni, M., Mazzucchelli, L., Martinelli, G., Pinotti, G., Ferreri, A., Doglioni, C., Zucca, E., Capella, C., Bertoni, F., Tibiletti, M, Martin, V, Bernasconi, B, Del Curto, B, Pecciarini, L, Uccella, S, Pruneri, G, Ponzoni, Maurilio, Mazzucchelli, L, Martinelli, G, Pinotti, G, Ferreri, A, Doglioni, Claudio, Zucca, E, Capella, C, and Bertoni, F.
- Published
- 2008
7. Immunoglobulin Gene Repertoire in Ocular Adnexa Lymphomas (OAL): Hints on the Nature of the Antigenic Stimulation
- Author
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Dagklis A, Ponzoni M, Ferreri AJM, Giulia M, Pecciarini L, Dolcetti R, Stamatopoulos K, Doglioni C, Ghia P, Dagklis, A, Ponzoni, M, Ferreri, Ajm, Giulia, M, Pecciarini, L, Dolcetti, R, Stamatopoulos, K, Doglioni, C, and Ghia, P
- Published
- 2008
8. CHLAMYDOPHILA PSITTACI (CP) IS VIABLE AND INFECTIOUS IN THE CONJUNCTIVA AND PERIPHERAL BLOOD OF PATIENTS WITH OCULAR ADNEXAL MALT LYMPHOMA (OAML): RESULTS OF A PROSPECTIVE CASE-CONTROL STUDY
- Author
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Ferreri, J. M., Riccardo Dolcetti, Dognini, P., Malabarba, L., Vicari, N., Pasini, E., Ponzoni, M., Cangi, M. G., Pecciarini, L., Resti, A. Giordano, Rossini, S., Doglioni, C., Magnino, S., Ferreri, Jm, Dolcetti, R, Dognini, P, Malabarba, L, Vicari, N, Pasini, E, Ponzoni, Maurilio, Cangi, Mg, Pecciarini, L, Resti, Ag, Rossini, S, Doglioni, C, and Magnino, S.
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- 2008
9. Prevalence of Chlamydia psittaci infection in nodal and extranodal non-Hodgkin lymphomas
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Ferreri, J. M., Dolcetti, R., Guidoboni, M., Maria Giulia Cangi, Pecciarini, L., Dognini, G. P., Ghia, P. P., Malnati, M., Pasini, E., Doglioni, C., Ponzoni, M., Ferreri, Jm, Dolcetti, R, Guidoboni, M, Cangi, Mg, Pecciarini, L, Dognini, Gp, Ghia, Pp, Malnati, M, Pasini, E, Doglioni, C, and Ponzoni, M
- Published
- 2007
10. Characterization of t(6,11)(p21;q12) in renal cell carcinoma of an adult patient
- Author
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Pecciarini, L, Cangi, M, LO CUNSOLO, C, Macri', E, DAL CIN, E, Martignoni, Guido, and Doglioni, C.
- Published
- 2007
11. Carcinomi con traslocazioni coinvolgenti i geni della famiglia MiTF/TFE
- Author
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Gobbo, S, Tardanico, R, Martignoni, Guido, Pea, Maurizio, Pecciarini, L, Brunelli, Matteo, Balzarini, P, Macri, E, Cossu Rocca, P, Chilosi, Marco, Menestrina, Fabio, and Doglioni, C.
- Published
- 2005
12. Neuroendocrinology of human placenta
- Author
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Petraglia, F., Florio, P., Luisi, S., Severi, F. M., Bocchi, C., Torricelli, M., Cobellis, L., Centini, G., Guidoni, G., Pecciarini, L., Cito, G., and Picciolini, E.
- Published
- 2003
13. Inhibins and activins in pregnancy
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Petraglia, F., Florio, P., Luisi, S., Severi, F. M., Ciarmela, P., Picciolini, E., Pecciarini, L., Cobellis, L., Bocchi, C., Bagnoli, F., and Buonocore, G.
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- 2003
14. p53-p63 cross-talk in lymphomas
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Doglioni, C, Cangi, G, Pecciarini, L, Zamo', Alberto, Canal, F, and Chilosi, Marco
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- 2002
15. Absent C-reactive response in early-onset neonatal sepsi. Relation to clinical characteristics and outcome
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De Felice, C., Liberatori, S., Bini, Luca, Stabile, M. E., Magi, Barbara, Marzocchi, Barbara, Pecciarini, L., Pallini, V., and Bracci, R.
- Published
- 1997
16. Estroprogestinici e mammella: indicazioni e controindicazioni nelle adolescenti
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Franchi, F, Vanni, A, Funicelli, V, Cresti, M, Giovani, M, Pecciarini, L, and Gioffre', WALTER RENATO
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- 1992
17. A Woman and Her Canary: A Tale of Chlamydiae and Lymphomas
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Ferreri, A. J. M., primary, Dolcetti, R., additional, Magnino, S., additional, Doglioni, C., additional, Cangi, M. G., additional, Pecciarini, L., additional, Ghia, P., additional, Dagklis, A., additional, Pasini, E., additional, Vicari, N., additional, Dognini, G. P., additional, Resti, A. G., additional, and Ponzoni, M., additional
- Published
- 2007
- Full Text
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18. A screening study of Chlamydia psittaci infection in 172 cases of nodal and extranodal non-hodgkin lymphomas
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Ferren, A. J. M., Dolcetti, R., Guidoboni, M., Maria Giulia Cangi, Pecciarini, L., Dognini, G. P., Ghia, P., Malnati, M., Pasini, E., Doglioni, C., Ponzoni, M., Ferren, Ajm, Dolcetti, R, Guidoboni, M, Cangi, Mg, Pecciarini, L, Dognini, Gp, Ghia, P, Malnati, M, Pasini, E, Doglioni, C, and Ponzoni, M
19. Malformazioni fetali riscontrate con ultrasuoni in un solo gemello in corso di gravidanze plurime. Considerazioni medico legali su 4 casi [Fetal malformations detected by ultrasonics in a twin in multiple pregnancies. Medico-legal considerations on 4 cases]
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Norelli, G. A., Centini, G., Calvelli, P., Pecciarini, L., Monti, E., and Severi, F. M.
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- 1988
20. Range di normalità dell'alpha-fetoproteina nel liquido amniotico e nel plasma materno [Normal range of alpha-fetoprotein in amniotic fluid and maternal plasma]
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Monti, E., gabriele centini, Landi, D., Pecciarini, L., Pasqui, L., and Severi, Filiberto Maria
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- 1987
21. Establishment and Characterization of PCL12, a Novel CD5+ Chronic Lymphocytic Leukaemia Cell Line
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Pamela Ranghetti, Maria Teresa Sabrina Bertilaccio, Gabriella Leone, Benedetta Apollonio, Federica Barbaglio, Cristina Scielzo, Maurilio Ponzoni, Lydia Scarfò, Andreas Agathangelidis, Paolo Ghia, Lorenza Pecciarini, Valeria De Pascali, Federico Caligaris-Cappio, Agathangelidis, A, Scarfò, L, Barbaglio, F, Apollonio, B, Bertilaccio, Mt, Ranghetti, P, Ponzoni, M, Leone, G, De Pascali, V, Pecciarini, L, Ghia, PAOLO PROSPERO, Caligaris Cappio, F, Scielzo, C., Agathangelidis, A., Scarfo', L., Barbaglio, F., Apollonio, B., Bertilaccio, M. T., Ranghetti, P., Ponzoni, M., Leone, G., De Pascali, V., Pecciarini, L., Ghia, P., and Caligaris-Cappio, F.
- Subjects
Chronic lymphocytic leukemia ,lcsh:Medicine ,Biology ,CD5 Antigens ,CD19 ,Mice ,immune system diseases ,Cell Line, Tumor ,hemic and lymphatic diseases ,Calcium flux ,medicine ,Animals ,Humans ,lcsh:Science ,Gene Rearrangement ,CD20 ,ZAP-70 Protein-Tyrosine Kinase ,Multidisciplinary ,ZAP70 ,lcsh:R ,hemic and immune systems ,Gene rearrangement ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Phenotype ,biology.protein ,Cancer research ,lcsh:Q ,CD5 ,Immortalised cell line ,Neoplasm Transplantation ,Research Article - 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.
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- 2015
22. 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.
- Subjects
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.
- Published
- 2021
23. 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.
- Subjects
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.
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- 2020
24. 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.
- Subjects
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.
- Published
- 2019
25. 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.
- Subjects
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.
- Published
- 2018
26. 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
- Subjects
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.
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- 2018
27. 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
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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
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- 2016
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
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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
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- 2016
29. 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
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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.
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- 2011
30. 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.
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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.
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- 2008
31. 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
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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.
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- 2007
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
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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. 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.
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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.
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- 2012
34. 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
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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.
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- 2008
35. Constitutive overexpression of CDC25A in primary human mammary epithelial cells results in both defective DNA damage response and chromosomal breaks at fragile sites
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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
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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.
- Published
- 2008
36. Chlamydia-psittaci-eradicating antibiotic therapy in patients with advanced-stage ocular adnexal MALT lymphoma
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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
- 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
37. A transcription-dependent micrococcal nuclease-resistant fragment of the urokinase-type plasminogen activator promoter interacts with the enhancer
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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
- 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.
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- 2007
38. A woman and her canary: a tale of chlamydiae and lymphomas
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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
- Subjects
Cancer Research ,Oncology ,biology ,Chlamydiales ,Immunology ,Chlamydophila infections ,medicine ,Chlamydiae ,Second primary cancer ,medicine.disease ,biology.organism_classification ,Virology ,Lymphoma - Published
- 2007
39. High syndecan-1 expression in breast carcinoma is related to an aggressive phenotype and to poorer prognosis
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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
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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
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- 2003
40. p63, a p53 homologue, is a selective nuclear marker of myoepithelial cells of the human breast
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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
- 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
- Published
- 2001
41. Routine Molecular Profiling in Both Resectable and Unresectable Pancreatic Adenocarcinoma: Relevance of Cytologic Samples.
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Redegalli M, Grassini G, Magliacane G, Pecciarini L, Schiavo Lena M, Smart CE, Johnston RL, Waddell N, Maestro R, Macchini M, Orsi G, Petrone MC, Rossi G, Balzano G, Falconi M, Arcidiacono PG, Reni M, Doglioni C, and Cangi MG
- Subjects
- Humans, Endoscopic Ultrasound-Guided Fine Needle Aspiration, Pancreatic Neoplasms, Pancreatic Neoplasms diagnosis, Pancreatic Neoplasms genetics, Pancreatic Neoplasms surgery, Adenocarcinoma diagnosis, Adenocarcinoma genetics, Adenocarcinoma surgery, Carcinoma, Pancreatic Ductal diagnosis, Carcinoma, Pancreatic Ductal genetics, Carcinoma, Pancreatic Ductal surgery
- Abstract
Background & Aims: 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., Methods: 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., Results: 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., Conclusions: 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., (Copyright © 2023 AGA Institute. Published by Elsevier Inc. All rights reserved.)
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- 2023
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42. Gene Fusion Detection in NSCLC Routine Clinical Practice: Targeted-NGS or FISH?
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Pecciarini L, Brunetto E, Grassini G, De Pascali V, Ogliari FR, Talarico A, Marra G, Magliacane G, Redegalli M, Arrigoni G, Lazzari C, Gregorc V, Bulotta A, Doglioni C, and Cangi MG
- Subjects
- Humans, Anaplastic Lymphoma Kinase genetics, In Situ Hybridization, Fluorescence methods, Receptor Protein-Tyrosine Kinases genetics, RNA therapeutic use, Gene Fusion genetics, Lung Neoplasms pathology, Carcinoma, Non-Small-Cell Lung pathology, Adenocarcinoma of Lung
- 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.
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- 2023
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43. Locally Performed HRD Testing for Ovarian Cancer? Yes, We Can!
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Magliacane G, Brunetto E, Calzavara S, Bergamini A, Pipitone GB, Marra G, Redegalli M, Grassini G, Rabaiotti E, Taccagni G, Pecciarini L, Carrera P, Mangili G, Doglioni C, and Cangi MG
- 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.
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- 2022
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44. Safety and efficacy of a dose-dense short-term therapy in patients with MYC-translocated aggressive lymphoma.
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Ferreri AJM, Angelillo P, Erbella F, Cattaneo C, Verga L, Lleshi A, Allione B, Ponzoni M, Facchetti F, Pagani C, Foppoli M, Pecciarini L, Sassone M, Steffanoni S, Flospergher E, Rossi G, Spina M, and Re A
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- Humans, Rituximab therapeutic use, Vincristine adverse effects, Etoposide adverse effects, Retrospective Studies, In Situ Hybridization, Fluorescence, Antibodies, Monoclonal, Murine-Derived, Antineoplastic Combined Chemotherapy Protocols adverse effects, Transplantation, Autologous, Cyclophosphamide adverse effects, Prednisone therapeutic use, Cytarabine adverse effects, Doxorubicin adverse effects, Hematopoietic Stem Cell Transplantation, COVID-19, Burkitt Lymphoma drug therapy, Burkitt Lymphoma genetics, Lymphoma, B-Cell drug therapy, Lymphoma drug therapy, HIV Infections drug therapy
- 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., (© 2022 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.)
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- 2022
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45. Case report: EML4::NTRK3 gene fusion in a patient with metastatic lung adenocarcinoma successfully treated with entrectinib.
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Lazzari C, Pecciarini L, Doglioni C, Pedica F, Gajate AMS, Bulotta A, Gregorc V, and Cangi MG
- Abstract
Rearrangements involving the neurotrophin kinase ( NTRK ) genes NTRK1 , NTRK2 and NTRK3 with different fusion partners have been observed in both adult and pediatric solid tumors. Larotrectinib and entrectinib have been the first tumor-agnostic compounds approved for the treatment of NTRK fusion-positive tumors. Here, we report the first case of a female patient with a diagnosis of stage IV lung adenocarcinoma harboring the EML4::NTRK3 gene fusion, and successfully treated with entrectinib., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Lazzari, Pecciarini, Doglioni, Pedica, Gajate, Bulotta, Gregorc and Cangi.)
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- 2022
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46. Intratumoral Cellular Heterogeneity: Implications for Drug Resistance in Patients with Non-Small Cell Lung Cancer.
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Gregorc V, Lazzari C, Mandalá M, Ippati S, Bulotta A, Cangi MG, Khater A, Viganò MG, Mirabile A, Pecciarini L, Ogliari FR, Arrigoni G, Grassini G, Veronesi G, and Doglioni C
- 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.
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- 2021
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47. Next Generation Sequencing in Non-Small Cell Lung Cancer: Pitfalls and Opportunities.
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Lazzari C, Bulotta A, Cangi MG, Bucci G, Pecciarini L, Bonfiglio S, Lorusso V, Ippati S, Arrigoni G, Grassini G, Doglioni C, and Gregorc V
- 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.
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- 2020
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48. Novel insights into the genetics and epigenetics of MALT lymphoma unveiled by next generation sequencing analyses.
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Cascione L, Rinaldi A, Bruscaggin A, Tarantelli C, Arribas AJ, Kwee I, Pecciarini L, Mensah AA, Spina V, Chung EYL, di Bergamo LT, Dirnhofer S, Tzankov A, Miranda RN, Young KH, Traverse-Glehen A, Gaidano G, Swerdlow SH, Gascoyne R, Rabadan R, Ponzoni M, Bhagat G, Rossi D, Zucca E, and Bertoni F
- Subjects
- Humans, Lymphoma, B-Cell, Marginal Zone classification, Prognosis, Biomarkers, Tumor genetics, Epigenesis, Genetic, Gene Expression Regulation, Neoplastic, High-Throughput Nucleotide Sequencing methods, Lymphoma, B-Cell, Marginal Zone genetics, Lymphoma, B-Cell, Marginal Zone pathology
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- 2019
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49. Modeling multiple myeloma-bone marrow interactions and response to drugs in a 3D surrogate microenvironment.
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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, and Ferrero E
- Subjects
- Bioreactors, Bortezomib pharmacology, Cell Adhesion, Cell Survival, Coculture Techniques, Drug Resistance, Endothelial Cells, Gelatin, Humans, Multiple Myeloma drug therapy, Stromal Cells, Tumor Microenvironment, Bone Marrow pathology, Cell Communication, Cell Culture Techniques, Models, Biological, Multiple Myeloma pathology
- 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., (Copyright© 2018 Ferrata Storti Foundation.)
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- 2018
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50. Establishment, characterization and long-term culture of human endocrine pancreas-derived microvascular endothelial cells.
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Sordi V, Ferri A, Ceserani V, Ciusani E, Dugnani E, Pellegrini S, Nano R, Pecciarini L, Pessina A, Pascucci L, Piemonti L, and Alessandri G
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- Antigens, CD metabolism, Cadherins metabolism, Cells, Cultured, Endothelial Cells cytology, Humans, Interleukin-8 metabolism, Microvessels cytology, Vascular Endothelial Growth Factor A metabolism, Vascular Endothelial Growth Factor Receptor-1, von Willebrand Factor metabolism, Endothelial Cells metabolism, Endothelium, Vascular cytology, Islets of Langerhans cytology
- 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., (Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
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