50 results on '"Vallacchi, V"'
Search Results
2. miR-146a-5p impairs melanoma resistance to kinase inhibitors by targeting COX2 and regulating NFkB-mediated inflammatory mediators
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Vergani, E, Dugo, M, Cossa, M, Frigerio, S, Di Guardo, L, Gallino, G, Mattavelli, I, Vergani, B, Lalli, L, Tamborini, E, Valeri, B, Gargiuli, C, Shahaj, E, Ferrarini, M, Ferrero, E, Gomez Lira, M, Huber, V, Vecchio, M, Sensi, M, Leone, B, Santinami, M, Rivoltini, L, Rodolfo, M, Vallacchi, V, Vergani E., Dugo M., Cossa M., Frigerio S., Di Guardo L., Gallino G., Mattavelli I., Vergani B., Lalli L., Tamborini E., Valeri B., Gargiuli C., Shahaj E., Ferrarini M., Ferrero E., Gomez Lira M., Huber V., Vecchio M. D., Sensi M., Leone B. E., Santinami M., Rivoltini L., Rodolfo M., Vallacchi V., Vergani, E, Dugo, M, Cossa, M, Frigerio, S, Di Guardo, L, Gallino, G, Mattavelli, I, Vergani, B, Lalli, L, Tamborini, E, Valeri, B, Gargiuli, C, Shahaj, E, Ferrarini, M, Ferrero, E, Gomez Lira, M, Huber, V, Vecchio, M, Sensi, M, Leone, B, Santinami, M, Rivoltini, L, Rodolfo, M, Vallacchi, V, Vergani E., Dugo M., Cossa M., Frigerio S., Di Guardo L., Gallino G., Mattavelli I., Vergani B., Lalli L., Tamborini E., Valeri B., Gargiuli C., Shahaj E., Ferrarini M., Ferrero E., Gomez Lira M., Huber V., Vecchio M. D., Sensi M., Leone B. E., Santinami M., Rivoltini L., Rodolfo M., and Vallacchi V.
- Abstract
Background: Targeted therapy with BRAF and MEK inhibitors has improved the survival of patients with BRAF-mutated metastatic melanoma, but most patients relapse upon the onset of drug resistance induced by mechanisms including genetic and epigenetic events. Among the epigenetic alterations, microRNA perturbation is associated with the development of kinase inhibitor resistance. Here, we identified and studied the role of miR-146a-5p dysregulation in melanoma drug resistance. Methods: The miR-146a-5p-regulated NFkB signaling network was identified in drug-resistant cell lines and melanoma tumor samples by expression profiling and knock-in and knock-out studies. A bioinformatic data analysis identified COX2 as a central gene regulated by miR-146a-5p and NFkB. The effects of miR-146a-5p/COX2 manipulation were studied in vitro in cell lines and with 3D cultures of treatment-resistant tumor explants from patients progressing during therapy. Results: miR-146a-5p expression was inversely correlated with drug sensitivity and COX2 expression and was reduced in BRAF and MEK inhibitor-resistant melanoma cells and tissues. Forced miR-146a-5p expression reduced COX2 activity and significantly increased drug sensitivity by hampering prosurvival NFkB signaling, leading to reduced proliferation and enhanced apoptosis. Similar effects were obtained by inhibiting COX2 by celecoxib, a clinically approved COX2 inhibitor. Conclusions: Deregulation of the miR-146a-5p/COX2 axis occurs in the development of melanoma resistance to targeted drugs in melanoma patients. This finding reveals novel targets for more effective combination treatment. [MediaObject not available: see fulltext.] Graphical Abstract: [Figure not available: see fulltext.]
- Published
- 2020
3. 3D tumor explant as a novel platform to investigate therapeutic pathways and predictive biomarkers in cancer patients
- Author
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Rodolfo, M, Huber, V, Cossa, M, Gallino, G, Leone, B, Vallacchi, V, Rivoltini, L, Vergani, E, Rodolfo, Monica, Huber, Veronica, Cossa, Mara, Gallino, Gianfrancesco, Leone, Biagio E, Vallacchi, Viviana, Rivoltini, Licia, Vergani, Elisabetta, Rodolfo, M, Huber, V, Cossa, M, Gallino, G, Leone, B, Vallacchi, V, Rivoltini, L, Vergani, E, Rodolfo, Monica, Huber, Veronica, Cossa, Mara, Gallino, Gianfrancesco, Leone, Biagio E, Vallacchi, Viviana, Rivoltini, Licia, and Vergani, Elisabetta
- Abstract
Immunotherapy with immune checkpoint inhibitors can induce durable clinical responses in different human malignancies but the number of responding patients remains globally modest. The limited therapeutic efficacy of ICI depends on multiple factors, among which the immune suppressive features of the tumor microenvironment play a key role. For this reason, experimental models that enable dissection of the immune-hostile tumor milieu components are required to unravel how to overcome resistance and obtain full-fledged anti-tumor immunity. Recent evidence supports the usefulness of 3D ex vivo systems in retaining features of tumor microenvironment to elucidate molecular and immunologic mechanisms of response and resistance to immune checkpoint blockade. In this perspective article we discuss the recent advances in patient-derived 3D tumor models and their potential in support of treatment decision making in clinical setting. We will also share our experience with dynamic bioreactor tumor explant culture of samples from melanoma and sarcoma patients as a reliable and promising platform to unravel immune responses to immune checkpoint inhibitors.
- Published
- 2022
4. Is impaired response to PD-1 blockers of high serum PD-1 patients related to immune complexes?
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Daveri, E., Luison, E., Vallacchi, V., Vergani, B., Leone, B.E., Garassino, M.C., Figini, M., and Rivoltini, L.
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- 2021
- Full Text
- View/download PDF
5. 1537P Dissecting inflammation in patients with desmoid fibromatosis to identify prognostic biomarkers
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Vallacchi, V., primary, Bergamaschi, L., additional, Perrone, F., additional, Rini, F., additional, Rivoltini, L., additional, Castelli, C., additional, Gronchi, A., additional, and Colombo, C., additional
- Published
- 2021
- Full Text
- View/download PDF
6. 970P Assessment of anti-PD-1 antibody and immune complex binding to Fcγ receptors and clinical implications
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Daveri, E., primary, Luison, E., additional, Vallacchi, V., additional, Vergani, B., additional, Leone, B.E., additional, Garassino, M.C., additional, Figini, M., additional, and Rivoltini, L., additional
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- 2021
- Full Text
- View/download PDF
7. The density and spatial tissue distribution of CD8(+) and CD163(+) immune cells predict response and outcome in melanoma patients receiving MAPK inhibitors
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Massi, D., Rulli, E., Cossa, M., Valeri, B., Rodolfo, M., Merelli, B., De Logu, F., Nassini, R., Del Vecchio, M., Di Guardo, L., De Penni, R., Guida, M., Sileni, V. C., Di Giacomo, A. M., Tucci, M., Occelli, M., Portelli, F., Vallacchi, V., Consoli, F., Quaglino, P., Queirolo, P., Baroni, G., Carnevale-Schianca, F., Cattaneo, L., Minisini, A., Palmieri, G., Rivoltini, L., Mandala, M., Simi, S., and Galli, F.
- Subjects
Male ,0301 basic medicine ,Oncology ,Cancer Research ,Kaplan-Meier Estimate ,B7-H1 Antigen ,0302 clinical medicine ,Receptors ,Tumor Microenvironment ,Immunology and Allergy ,Melanoma ,beta Catenin ,Middle Aged ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,MAP Kinase Kinase Kinases ,CD ,3. Good health ,Differentiation ,030220 oncology & carcinogenesis ,Cell Surface ,Myeloid cells ,Melanoma prognosis ,Microenvironment ,T lymphocytes ,Aged ,Antigens, CD ,Antigens, Differentiation, Myelomonocytic ,CD8 Antigens ,Female ,Humans ,Programmed Cell Death 1 Ligand 2 Protein ,Protein Kinase Inhibitors ,Proto-Oncogene Proteins B-raf ,Receptors, Cell Surface ,Molecular Medicine ,Research Article ,medicine.medical_specialty ,Immunology ,lcsh:RC254-282 ,03 medical and health sciences ,Immune system ,Immunity ,Internal medicine ,medicine ,Progression-free survival ,Antigens ,Pharmacology ,Tumor microenvironment ,Performance status ,business.industry ,Myelomonocytic ,medicine.disease ,030104 developmental biology ,business ,CD163 ,CD8 - Abstract
Background Clinical response to MAPK inhibitors in metastatic melanoma patients is heterogeneous for reasons still needing to be elucidated. As the patient immune activity contributes to treatment clinical benefit, the pre-existing level of immunity at tumor site may provide biomarkers of disease outcome to therapy. Here we investigated whether assessing the density and spatial tissue distribution of key immune cells in the tumor microenvironment could identify patients predisposed to respond to MAPK inhibitors. Methods Pretreatment tumor biopsies from a total of 213 patients (158 for the training set and 55 for the validation set) treated with BRAF or BRAF/MEK inhibitors within the Italian Melanoma Intergroup were stained with selected immune markers (CD8, CD163, beta-catenin, PD-L1, PD-L2). Results, obtained by blinded immunohistochemical scoring and digital image analysis, were correlated with clinical response and outcome by multivariate logistic models on response to treatment and clinical outcome, adjusted for American Joint Committee on Cancer stage, performance status, lactate dehydrogenase and treatment received. Results Patients with high intratumoral, but not peritumoral, CD8(+) T cells and concomitantly low CD163(+) myeloid cells displayed higher probability of response (OR 9.91, 95% CI 2.23-44.0, p = 0.003) and longer overall survival (HR 0.34, 95% CI 0.16-0.72, p = 0.005) compared to those with intratumoral low CD8(+) T cells and high CD163(+) myeloid cells. The latter phenotype was instead associated with a shorter progression free survival (p = 0.010). In contrast, PD-L1 and PD-L2 did not correlate with clinical outcome while tumor beta-catenin overexpression showed association with lower probability of response (OR 0.48, 95% CI 0.21-1.06, p = 0.068). Conclusions Analysis of the spatially constrained distribution of CD8(+) and CD163(+) cells, representative of the opposite circuits of antitumor vs protumor immunity, respectively, may assist in identifying melanoma patients with improved response and better outcome upon treatment with MAPK inhibitors. These data underline the role of endogenous immune microenvironment in predisposing metastatic melanoma patients to benefit from therapies targeting driver-oncogenic pathways.
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- 2019
- Full Text
- View/download PDF
8. Overcoming melanoma resistance to vemurafenib by targeting CCL2-induced miR-34a, miR-100 and miR-125b
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Vergani, E, Di Guardo, L, Dugo, M, Rigoletto, S, Tragni, G, Ruggeri, R, Perrone, F, Tamborini, E, Gloghini, A, Arienti, F, Vergani, B, Deho, P, De Cecco, L, Vallacchi, V, Frati, P, Shahaj, E, Villa, A, Santinami, M, De Braud, F, Rivoltini, L, Rodolfo, M, Rodolfo, M., VERGANI, BARBARA, VILLA, ANTONELLO, Vergani, E, Di Guardo, L, Dugo, M, Rigoletto, S, Tragni, G, Ruggeri, R, Perrone, F, Tamborini, E, Gloghini, A, Arienti, F, Vergani, B, Deho, P, De Cecco, L, Vallacchi, V, Frati, P, Shahaj, E, Villa, A, Santinami, M, De Braud, F, Rivoltini, L, Rodolfo, M, Rodolfo, M., VERGANI, BARBARA, and VILLA, ANTONELLO
- Abstract
In melanoma, the adaptative cell response to BRAF inhibitors includes altered patterns of cytokine production contributing to tumor progression and drug resistance. Among the factors produced by PLX4032-resistant melanoma cell lines, CCL2 was higher compared to the sensitive parental cell lines and increased upon drug treatment. CCL2 acted as an autocrine growth factor for melanoma cells, stimulating the proliferation and resistance to apoptosis. In patients, CCL2 is detected in melanoma cells in tumors and in plasma at levels that correlate with tumor burden and lactate dehydrogenase. Vemurafenib treatment increased the CCL2 levels in plasma, whereas the long-term clinical response was associated with low CCL2 levels. Increased CCL2 production was associated with miRNA deregulation in the resistant cells. miR-34a, miR-100 and miR-125b showed high expression in both resistant cells and in tumor biopsies that were obtained from treated patients, and they were involved in the control of cell proliferation and apoptosis. Inhibition of CCL2 and of the selected miRNAs restored both the cell apoptosis and the drug efficacy in resistant melanoma cells. Therefore, CCL2 and miRNAs are potential prognostic factors and attractive targets for counteracting treatment resistance in metastatic melanoma.
- Published
- 2016
9. Tenth annual meeting of the Italian Network for Tumor Biotherapy (NIBIT), SIENA, Italy, November 5-7, 2012
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Maio, M., Nicolay, H. J. M., Ascierto, P. A., Belardelli, F., Camerini, R., Colombo, M. P., Queirolo, P., Ridolfi, R., Russo, V., Parisi, G., Cutaia, O., Fonsatti, E., Parmiani, G., Mennonna, D., Carluccio, S., Bellone, M., Maccalli, C., Brendolan, A., Mondino, A., Corti, A., Bondanza, A., Locatelli, F., Seliger, B., Filaci, G., Rosato, A., Pittoni, P., Tazzari, M., Rivoltini, L., Anichini, A., Vallacchi, V., Zappasodi, R., Quaglino, E., Moresco, R. M., Camisaschi, C., Calabro, L., Ferrucci, P. F., Fratta, E., Ugel, S., van Baren, N., Guidoboni, M., Covre, A., Carbone, E., Aurisicchio, L., Palmieri, G., Di Giacomo, A. M., Volonte, A., Jachetti, E., and Sangiolo, D.
- Subjects
Cancer Research ,medicine.medical_specialty ,business.industry ,Immunology ,Cancer ,medicine.disease ,NIBIT ,Oncology ,Family medicine ,Immunotherapy ,Networks ,Cancer, Immunology, Immunotherapy, Networks, NIBIT ,Immunology and Allergy ,Medicine ,business - Published
- 2013
10. Transcriptional profiling of melanoma sentinel nodes identify patients with poor outcome and reveal an association of CD30(+) T lymphocytes with progression
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Vallacchi, V, Vergani, E, Camisaschi, C, Deho, P, Cabras, A, Sensi, M, De Cecco, L, Bassani, N, Ambrogi, F, Carbone, A, Crippa, F, Vergani, B, Frati, P, Arienti, F, Patuzzo, R, Villa, A, Biganzoli, E, Canevari, S, Santinami, M, Castelli, C, Rivoltini, L, Rodolfo, M, VERGANI, BARBARA, VILLA, ANTONELLO, Rodolfo, M., Vallacchi, V, Vergani, E, Camisaschi, C, Deho, P, Cabras, A, Sensi, M, De Cecco, L, Bassani, N, Ambrogi, F, Carbone, A, Crippa, F, Vergani, B, Frati, P, Arienti, F, Patuzzo, R, Villa, A, Biganzoli, E, Canevari, S, Santinami, M, Castelli, C, Rivoltini, L, Rodolfo, M, VERGANI, BARBARA, VILLA, ANTONELLO, and Rodolfo, M.
- Abstract
Sentinel lymph nodes set the stance of the immune system to a localized tumor and are often the first site to be colonized by neoplastic cells that metastasize. To investigate how the presence of neoplastic cells in sentinel lymph nodes may trigger pathways associated with metastatic progression, we analyzed the transcriptional profiles of archival sentinel node biopsy specimens obtained from melanoma patients. Biopsies from positive nodes were selected for comparable tumor infiltration, presence or absence of further regional node metastases, and relapse at 5-year follow-up. Unsupervised analysis of gene expression profiles revealed immune response to be a major gene ontogeny represented. Among genes upregulated in patients with progressing disease, the TNF receptor family member CD30/TNFRSF8 was confirmed in biopsy specimens from an independent group of patients. Immunohistochemical analysis revealed higher numbers of CD30(+) lymphocytes in nodes from progressing patients compared with nonprogressing patients. Phenotypic profiling demonstrated that CD30(+) lymphocytes comprised a broad population of suppressive or exhausted immune cells, such as CD4(+)Foxp3(+) or PD1(+) subpopulations and CD4(-)CD8(-) T cells. CD30(+) T lymphocytes were increased in peripheral blood lymphocytes of melanoma patients at advanced disease stages. Our findings reinforce the concept that sentinel nodes act as pivotal sites for determining progression patterns, revealing that the presence of CD30(+) lymphocytes at those sites associate positively with melanoma progression.
- Published
- 2014
11. Molecular profiling of the 'plexinome' in melanoma and pancreatic cancer
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Balakrishnan, A, Penachioni, Jy, Lamba, S, Bleeker, Fe, Zanon, C, Rodolfo, M, Vallacchi, V, Scarpa, A, Felicioni, L, Buck, M, Marchetti, A, Comoglio, Pm, Bardelli, A, Tamagnone, Luca, Tamagnone L (ORCID:0000-0002-2884-7946), Balakrishnan, A, Penachioni, Jy, Lamba, S, Bleeker, Fe, Zanon, C, Rodolfo, M, Vallacchi, V, Scarpa, A, Felicioni, L, Buck, M, Marchetti, A, Comoglio, Pm, Bardelli, A, Tamagnone, Luca, and Tamagnone L (ORCID:0000-0002-2884-7946)
- Abstract
Plexins are transmembrane high-affinity receptors for semaphorins, regulating cell guidance, motility, and invasion. Functional evidences implicate semaphorin signals in cancer progression and metastasis. Yet, it is largely unknown whether plexin genes are genetically altered in human tumors. We performed a comprehensive gene copy analysis and mutational profiling of all nine members of the plexin gene family (plexinome), in melanomas and pancreatic ductal adenocarcinomas (PDACs), which are characterized by high metastatic potential and poor prognosis. Gene copy analysis detected amplification of PLXNA4 in melanomas, whereas copy number losses of multiple plexin genes were seen in PDACs. Somatic mutations were detected in PLXNA4, PLXNB3, and PLXNC1; providing the first evidence that these plexins are mutated in human cancer. Functional assays in cellular models revealed that some of these missense mutations result in loss of plexin function. For instance, c.1613G>A, p.R538H mutation in the extracellular domain of PLXNB3 prevented binding of the ligand Sema5A. Moreover, although PLXNA4 signaling can inhibit tumor cell migration, the mutated c.5206C>T, p.H1736Y allele had lost this activity. Our study is the first systematic analysis of the "plexinome" in human tumors, and indicates that multiple mutated plexins may be involved in cancer progression.
- Published
- 2009
12. CCN3/NOV matricellular protein is expressed in melanoma progression
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Vallacchi, V., primary, Daniotti, M., additional, Ratti, F., additional, Perbal, B., additional, Parmiani, G., additional, and Rodolfo, M., additional
- Published
- 2006
- Full Text
- View/download PDF
13. Targeting Immune Regulatory Networks to Counteract Immune Suppression in Cancer
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Camisaschi C, Vallacchi V, Vergani E, Marcella Tazzari, Ferro S, Tuccitto A, Kuchuk O, Shahaj E, Sulsenti R, Castelli C, Rodolfo M, Rivoltini L, and Huber V
14. 3D tumor explant as a novel platform to investigate therapeutic pathways and predictive biomarkers in cancer patients
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Rodolfo, Monica, Huber, Veronica, Cossa, Mara, Gallino, Gianfrancesco, Leone, Biagio E., Vallacchi, Viviana, Rivoltini, Licia, Vergani, Elisabetta, Rodolfo, M, Huber, V, Cossa, M, Gallino, G, Leone, B, Vallacchi, V, Rivoltini, L, and Vergani, E
- Subjects
bioreactor ,sarcoma ,Immunology ,melanoma ,Immunology and Allergy ,immunotherapy ,patient-derived 3D tumor explant - Abstract
Immunotherapy with immune checkpoint inhibitors can induce durable clinical responses in different human malignancies but the number of responding patients remains globally modest. The limited therapeutic efficacy of ICI depends on multiple factors, among which the immune suppressive features of the tumor microenvironment play a key role. For this reason, experimental models that enable dissection of the immune-hostile tumor milieu components are required to unravel how to overcome resistance and obtain full-fledged anti-tumor immunity. Recent evidence supports the usefulness of 3D ex vivo systems in retaining features of tumor microenvironment to elucidate molecular and immunologic mechanisms of response and resistance to immune checkpoint blockade. In this perspective article we discuss the recent advances in patient-derived 3D tumor models and their potential in support of treatment decision making in clinical setting. We will also share our experience with dynamic bioreactor tumor explant culture of samples from melanoma and sarcoma patients as a reliable and promising platform to unravel immune responses to immune checkpoint inhibitors.
- Published
- 2022
15. miR-146a-5p impairs melanoma resistance to kinase inhibitors by targeting COX2 and regulating NFkB-mediated inflammatory mediators
- Author
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Elisabetta Ferrero, Marina Ferrarini, Chiara Gargiuli, Viviana Vallacchi, Michele Del Vecchio, Macarena Gomez Lira, Eriomina Shahaj, Mara Cossa, Elena Tamborini, Barbara Vergani, Luca Lalli, Mario Santinami, Barbara Valeri, Monica Rodolfo, Gianfrancesco Gallino, Simona Frigerio, Veronica Huber, Marialuisa Sensi, Licia Rivoltini, Elisabetta Vergani, Lorenza Di Guardo, Matteo Dugo, Ilaria Mattavelli, Biagio Eugenio Leone, Vergani, E, Dugo, M, Cossa, M, Frigerio, S, Di Guardo, L, Gallino, G, Mattavelli, I, Vergani, B, Lalli, L, Tamborini, E, Valeri, B, Gargiuli, C, Shahaj, E, Ferrarini, M, Ferrero, E, Gomez Lira, M, Huber, V, Vecchio, M, Sensi, M, Leone, B, Santinami, M, Rivoltini, L, Rodolfo, M, and Vallacchi, V
- Subjects
Proto-Oncogene Proteins B-raf ,BRAF/MEK inhibitors ,medicine.medical_treatment ,lcsh:Medicine ,Drug resistance ,Models, Biological ,Biochemistry ,Targeted therapy ,03 medical and health sciences ,0302 clinical medicine ,miR-146a-5p ,Cell Line, Tumor ,microRNA ,medicine ,Humans ,Epigenetics ,BRAF/MEK inhibitor ,lcsh:QH573-671 ,Extracellular Signal-Regulated MAP Kinases ,Melanoma ,Protein Kinase Inhibitors ,Molecular Biology ,030304 developmental biology ,Mitogen-Activated Protein Kinase Kinases ,0303 health sciences ,Kinase ,business.industry ,lcsh:Cytology ,Research ,lcsh:R ,NF-kappa B ,Cell Biology ,medicine.disease ,Gene Expression Regulation, Neoplastic ,Gene expression profiling ,MicroRNAs ,Cyclooxygenase 2 ,Drug Resistance, Neoplasm ,Apoptosis ,030220 oncology & carcinogenesis ,Cancer research ,Inflammation Mediators ,Melanoma resistance ,business ,Proto-Oncogene Proteins c-akt ,COX2 ,Signal Transduction - Abstract
Background Targeted therapy with BRAF and MEK inhibitors has improved the survival of patients with BRAF-mutated metastatic melanoma, but most patients relapse upon the onset of drug resistance induced by mechanisms including genetic and epigenetic events. Among the epigenetic alterations, microRNA perturbation is associated with the development of kinase inhibitor resistance. Here, we identified and studied the role of miR-146a-5p dysregulation in melanoma drug resistance. Methods The miR-146a-5p-regulated NFkB signaling network was identified in drug-resistant cell lines and melanoma tumor samples by expression profiling and knock-in and knock-out studies. A bioinformatic data analysis identified COX2 as a central gene regulated by miR-146a-5p and NFkB. The effects of miR-146a-5p/COX2 manipulation were studied in vitro in cell lines and with 3D cultures of treatment-resistant tumor explants from patients progressing during therapy. Results miR-146a-5p expression was inversely correlated with drug sensitivity and COX2 expression and was reduced in BRAF and MEK inhibitor-resistant melanoma cells and tissues. Forced miR-146a-5p expression reduced COX2 activity and significantly increased drug sensitivity by hampering prosurvival NFkB signaling, leading to reduced proliferation and enhanced apoptosis. Similar effects were obtained by inhibiting COX2 by celecoxib, a clinically approved COX2 inhibitor. Conclusions Deregulation of the miR-146a-5p/COX2 axis occurs in the development of melanoma resistance to targeted drugs in melanoma patients. This finding reveals novel targets for more effective combination treatment. Graphical Abstract
- Published
- 2020
16. Pazopanib for treatment of advanced extraskeletal myxoid chondrosarcoma: a multicentre, single-arm, phase 2 trial
- Author
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A. Lecesne, Dominga Racanelli, Jean-Yves Blay, Anna Maria Frezza, Marie Karanian, Silvia Brich, Daniel Bernabeu, Antonio Lopez-Pousa, María Ángeles Vaz Salgado, Paolo G. Casali, Sarah Dumont, Silvia Stacchiotti, Carlo Morosi, Chiara Castelli, Josefina Cruz, Monica Brenca, Antonio Gutierrez, Giovanni Grignani, Roberta Maestro, Nicolas Penel, Stefano Ferrari, Nadia Hindi, Andrés Redondo, Gianpaolo Dagrada, Javier Martin-Broto, Emanuela Palmerini, Paola Collini, Enrique de Álava, Antoine Italiano, Viviana Vallacchi, Grupo Español de Investigación en Sarcomas, Italian Sarcoma Group, French Sarcoma Group, GlaxoSmithKline, Novartis, Stacchiotti S., Ferrari S., Redondo A., Hindi-Muniz N., Palmerini E., Vaz Salgado M.A., Frezza A.M., Casali P.G., Gutierrez A., Lopez-Pousa A., Grignani G., Italiano A., LeCesne A., Dumont S., Blay J.Y., Penel N., Bernabeu D., de Alava E., Karanian M., Morosi C., Brich S., Dagrada G.P., Vallacchi V., Castelli C., Brenca M., Racanelli D., Maestro R., Collini P., Cruz J., and Martin-Broto J.
- Subjects
0301 basic medicine ,Oncology ,Male ,musculoskeletal diseases ,medicine.medical_specialty ,Indazoles ,animal structures ,Population ,Chondrosarcoma ,Soft Tissue Neoplasms ,Disease-Free Survival ,Pazopanib ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Clinical endpoint ,Humans ,education ,Aged ,Neoplasm Staging ,Retrospective Studies ,education.field_of_study ,Sulfonamides ,business.industry ,Retrospective cohort study ,Extraskeletal Myxoid Chondrosarcoma ,Middle Aged ,medicine.disease ,musculoskeletal system ,extraskeletal myxoid chondrosarcoma, pazopanib, target therapy, angiogenesis ,030104 developmental biology ,Pyrimidines ,Treatment Outcome ,Response Evaluation Criteria in Solid Tumors ,030220 oncology & carcinogenesis ,Cohort ,embryonic structures ,Female ,Sarcoma ,business ,Neoplasms, Connective and Soft Tissue ,medicine.drug - Abstract
[Background] Extraskeletal myxoid chondrosarcoma is a rare sarcoma with low sensitivity to cytotoxic chemotherapy. Retrospective evidence suggests that antiangiogenic drugs could be a treatment option. We aimed to investigate the activity of pazopanib, an antiangiogenic drug, in patients with advanced extraskeletal myxoid chondrosarcoma., [Methods] In this single-arm, open-label phase 2 trial, three parallel independent cohorts of different histotypes of advanced sarcomas were recruited (extraskeletal myxoid chondrosarcoma, typical solitary fibrous tumour, and malignant-dedifferentiated solitary fibrous tumour). In each cohort, patients received pazopanib. In this Article, we report the results of the cohort of patients with advanced extraskeletal myxoid chondrosarcoma. Separate reporting of the three cohorts was prespecified in the study protocol. In this cohort, adult patients (aged ≥18 years) with a diagnosis of NR4A3-translocated, metastatic, or unresectable extraskeletal myxoid chondrosarcoma, who had Response Evaluation Criteria in Solid Tumors (RECIST) progression in the previous 6 months, and had an Eastern Cooperative Oncology Group performance status of 0–2, were enrolled at 11 study sites of the Spanish, Italian, and French sarcoma groups. Patients received oral pazopanib (800 mg/day) continuously, until disease progression, unacceptable toxicity, death, non-compliance, patient refusal, or investigator's decision. The primary endpoint was the proportion of patients achieving an objective response according to RECIST 1·1 in the modified intention-to-treat population (patients who provided consent and had a central molecularly confirmed diagnosis of extraskeletal myxoid chondrosarcoma). The safety analysis included all patients who received at least one dose of pazopanib. This study is registered with ClinicalTrials.gov, number NCT02066285., [Findings] Between June 24, 2014, and Jan 17, 2017, 26 patients entered the study and started pazopanib. Of these, 23 met the eligibility criteria for the modified intention-to-treat analysis. Median follow-up was 27 months (IQR 18–30). 22 patients (one patient died before the primary analysis) were evaluable for the primary endpoint: four (18% [95% CI 1–36]) had a RECIST objective response. No deaths or grade 4 adverse events occurred. The most frequent grade 3 adverse events were hypertension (nine [35%] of 26 patients), increased concentration of alanine aminotransferase (six [23%]), and increased aspartate aminotransferase (five [19%])., [Interpretation] Pazopanib had clinically meaningful antitumour activity in patients with progressive and advanced extraskeletal myxoid chondrosarcoma, and could be considered a suitable option after failure to respond to first-line anthracycline-based chemotherapy in these patients., Funding: Spanish Group for Research on Sarcomas, Italian Sarcoma Group, French Sarcoma Group, GlaxoSmithKline, and Novartis.
- Published
- 2019
17. Transcriptional Profiling of Melanoma Sentinel Nodes Identify Patients with Poor Outcome and Reveal an Association of CD30+ T Lymphocytes with Progression
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Antonello Domenico Cabras, Paola Deho, Chiara Camisaschi, Monica Rodolfo, Barbara Vergani, Federica Crippa, Niccolò Bassani, Antonino Carbone, Roberto Patuzzo, Silvana Canevari, Paola Frati, Flavio Arienti, Mario Santinami, Loris De Cecco, Licia Rivoltini, Viviana Vallacchi, Elia Biganzoli, Federico Ambrogi, Chiara Castelli, Antonello Villa, Marialuisa Sensi, Elisabetta Vergani, Vallacchi, V, Vergani, E, Camisaschi, C, Deho, P, Cabras, A, Sensi, M, De Cecco, L, Bassani, N, Ambrogi, F, Carbone, A, Crippa, F, Vergani, B, Frati, P, Arienti, F, Patuzzo, R, Villa, A, Biganzoli, E, Canevari, S, Santinami, M, Castelli, C, Rivoltini, L, and Rodolfo, M
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,CD30 ,T-Lymphocytes ,Population ,Ki-1 Antigen ,Antigens, CD30 ,Immune system ,Biopsy ,medicine ,Humans ,education ,Melanoma ,education.field_of_study ,medicine.diagnostic_test ,Sentinel Lymph Node Biopsy ,business.industry ,Computational Biology ,FOXP3 ,Sentinel node ,medicine.disease ,Immunohistochemistry ,Treatment Outcome ,T-Lymphocyte ,Oncology ,Disease Progression ,Transcriptome ,business ,CD8 ,Human - Abstract
Sentinel lymph nodes set the stance of the immune system to a localized tumor and are often the first site to be colonized by neoplastic cells that metastasize. To investigate how the presence of neoplastic cells in sentinel lymph nodes may trigger pathways associated with metastatic progression, we analyzed the transcriptional profiles of archival sentinel node biopsy specimens obtained from melanoma patients. Biopsies from positive nodes were selected for comparable tumor infiltration, presence or absence of further regional node metastases, and relapse at 5-year follow-up. Unsupervised analysis of gene expression profiles revealed immune response to be a major gene ontogeny represented. Among genes upregulated in patients with progressing disease, the TNF receptor family member CD30/TNFRSF8 was confirmed in biopsy specimens from an independent group of patients. Immunohistochemical analysis revealed higher numbers of CD30+ lymphocytes in nodes from progressing patients compared with nonprogressing patients. Phenotypic profiling demonstrated that CD30+ lymphocytes comprised a broad population of suppressive or exhausted immune cells, such as CD4+Foxp3+ or PD1+ subpopulations and CD4−CD8− T cells. CD30+ T lymphocytes were increased in peripheral blood lymphocytes of melanoma patients at advanced disease stages. Our findings reinforce the concept that sentinel nodes act as pivotal sites for determining progression patterns, revealing that the presence of CD30+ lymphocytes at those sites associate positively with melanoma progression. Cancer Res; 74(1); 130–40. ©2014 AACR.
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- 2014
18. Overcoming melanoma resistance to vemurafenib by targeting CCL2-induced miR-34a, miR-100 and miR-125b
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Viviana Vallacchi, Flavio Arienti, Elena Tamborini, Monica Rodolfo, Paola Frati, Elisabetta Vergani, Paola Deho, Filippo de Braud, Licia Rivoltini, Loris De Cecco, Barbara Vergani, Federica Perrone, Mario Santinami, Roberta Ruggeri, Gabrina Tragni, Antonello Villa, Lorenza Di Guardo, Annunziata Gloghini, Matteo Dugo, Sara Rigoletto, Eriomina Shahaj, Vergani, E, Di Guardo, L, Dugo, M, Rigoletto, S, Tragni, G, Ruggeri, R, Perrone, F, Tamborini, E, Gloghini, A, Arienti, F, Vergani, B, Deho, P, De Cecco, L, Vallacchi, V, Frati, P, Shahaj, E, Villa, A, Santinami, M, De Braud, F, Rivoltini, L, and Rodolfo, M
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0301 basic medicine ,Male ,Indoles ,medicine.medical_treatment ,Drug resistance ,Tumor Cells, Cultured ,Medicine ,Vemurafenib ,Chemokine CCL2 ,Sulfonamides ,Reverse Transcriptase Polymerase Chain Reaction ,Melanoma ,Middle Aged ,Prognosis ,Gene Expression Regulation, Neoplastic ,Survival Rate ,Cytokine ,Oncology ,miRNAs ,Female ,MiRNA ,CCL2 ,medicine.drug ,Research Paper ,Adult ,BRAF inhibitor ,Blotting, Western ,Real-Time Polymerase Chain Reaction ,03 medical and health sciences ,microRNA ,melanoma ,Humans ,RNA, Messenger ,Aged ,Neoplasm Staging ,drug resistance ,business.industry ,Cell growth ,Gene Expression Profiling ,medicine.disease ,MicroRNAs ,030104 developmental biology ,Apoptosis ,Tumor progression ,Drug Resistance, Neoplasm ,Case-Control Studies ,Immunology ,Cancer research ,business - Abstract
In melanoma, the adaptative cell response to BRAF inhibitors includes altered patterns of cytokine production contributing to tumor progression and drug resistance. Among the factors produced by PLX4032-resistant melanoma cell lines, CCL2 was higher compared to the sensitive parental cell lines and increased upon drug treatment. CCL2 acted as an autocrine growth factor for melanoma cells, stimulating the proliferation and resistance to apoptosis. In patients, CCL2 is detected in melanoma cells in tumors and in plasma at levels that correlate with tumor burden and lactate dehydrogenase. Vemurafenib treatment increased the CCL2 levels in plasma, whereas the long-term clinical response was associated with low CCL2 levels. Increased CCL2 production was associated with miRNA deregulation in the resistant cells. miR-34a, miR-100 and miR-125b showed high expression in both resistant cells and in tumor biopsies that were obtained from treated patients, and they were involved in the control of cell proliferation and apoptosis. Inhibition of CCL2 and of the selected miRNAs restored both the cell apoptosis and the drug efficacy in resistant melanoma cells. Therefore, CCL2 and miRNAs are potential prognostic factors and attractive targets for counteracting treatment resistance in metastatic melanoma.
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- 2015
19. Is impaired response to PD-1 blockers of high serum PD-1 patients related to immune complexes?
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Marina Chiara Garassino, E. Luison, Biagio Eugenio Leone, Barbara Vergani, M. Figini, E. Daveri, V. Vallacchi, Licia Rivoltini, Daveri, E, Luison, E, Vallacchi, V, Vergani, B, Leone, B, Garassino, M, Figini, M, and Rivoltini, L
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business.industry ,Programmed Cell Death 1 Receptor ,High serum ,MEDLINE ,Neoplasms, Second Primary ,Antigen-Antibody Complex ,Hematology ,B7-H1 Antigen ,Immune system ,Text mining ,Oncology ,Immunology ,Humans ,Medicine ,business ,Melanoma ,Human - Full Text
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20. Circulating Tumor DNA in Patients with Desmoid Fibromatosis during Active Surveillance.
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Bergamaschi L, Zorza M, Rini F, Perrone F, Rivoltini L, Gronchi A, Pasquali S, Zaffaroni N, Vallacchi V, and Colombo C
- Abstract
Background: Sporadic desmoid fibromatosis (DF) is a rare locally aggressive tumor characterized by mutation in exon 3 of CTNNB1 (T41A, S45F, and S45P). Standard of care is active surveillance (AS), but 30% require treatment. DF clinical course is unpredictable and identification of prognostic markers is needed to tailor strategy. In this prospective study, we investigated the consistency between mutation detected in tumor biopsies with that detected in plasma by digital droplet PCR (ddPCR) and the association between circulating tumor DNA (ctDNA) abundancy with clinical outcome., Patients and Methods: A total of 56 patients and 10 healthy donors were included. CTNNB1 mutation status of DF biopsies was determined by Sanger and in case of WT CTNNB1 with NGS. In matched plasma samples at enrollment and during AS at specific timepoints, we evaluated cfDNA quantity and ctDNA., Results: ctDNA levels were measured in 46 patients with CTNNB1 mutation. Detection rate for T41A, S45F and S45P was 68%, 42% and 100%, respectively. S45P variant has been detected in all patients with S45P mutation. Longitudinal assessment of ctDNA during AS in nine patients (four with regression and five with progression as first event according to RECIST) showed a concordance between the event and ctDNA level change in six out of nine patients tested (4/5 with progression and 2/4 with regression)., Conclusions: Results of ctDNA analysis support its potential clinical implementation as diagnostic tool in specific clinical scenarios where biopsy can be challenging. A prospective clinical trial needs to be performed to evaluate the potential role of ctDNA as predictive biomarker., (© 2024. Society of Surgical Oncology.)
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- 2024
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21. Spatial distribution of tumour immune infiltrate predicts outcomes of patients with high-risk soft tissue sarcomas after neoadjuvant chemotherapy.
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Pasquali S, Vallacchi V, Lalli L, Collini P, Barisella M, Romagosa C, Bague S, Coindre JM, Dei Tos AP, Palmerini E, Quagliuolo V, Martin-Broto J, Lopez-Pousa A, Grignani G, Blay JY, Beveridge RD, Casiraghi E, Brich S, Renne SL, Bergamaschi L, Vergani B, Sbaraglia M, Casali PG, Rivoltini L, Stacchiotti S, and Gronchi A
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- Humans, Female, Male, Prognosis, Middle Aged, Adult, Aged, Treatment Outcome, Tumor Microenvironment immunology, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Biomarkers, Tumor, Immunohistochemistry, Neoadjuvant Therapy, Sarcoma drug therapy, Sarcoma mortality, Sarcoma immunology, Sarcoma pathology, Lymphocytes, Tumor-Infiltrating immunology, Lymphocytes, Tumor-Infiltrating metabolism
- Abstract
Background: Anthracycline-based neoadjuvant chemotherapy (NAC) may modify tumour immune infiltrate. This study characterized immune infiltrate spatial distribution after NAC in primary high-risk soft tissue sarcomas (STS) and investigate association with prognosis., Methods: The ISG-STS 1001 trial randomized STS patients to anthracycline plus ifosfamide (AI) or a histology-tailored (HT) NAC. Four areas of tumour specimens were sampled: the area showing the highest lymphocyte infiltrate (HI) at H&E; the area with lack of post-treatment changes (highest grade, HG); the area with post-treatment changes (lowest grade, LG); and the tumour edge (TE). CD3, CD8, PD-1, CD20, FOXP3, and CD163 were analyzed at immunohistochemistry and digital pathology. A machine learning method was used to generate sarcoma immune index scores (SIS) that predict patient disease-free and overall survival (DFS and OS)., Findings: Tumour infiltrating lymphocytes and PD-1+ cells together with CD163+ cells were more represented in STS histologies with complex compared to simple karyotype, while CD20+ B-cells were detected in both these histology groups. PD-1+ cells exerted a negative prognostic value irrespectively of their spatial distribution. Enrichment in CD20+ B-cells at HI and TE areas was associated with better patient outcomes. We generated a prognostic SIS for each tumour area, having the HI-SIS the best performance. Such prognostic value was driven by treatment with AI., Interpretation: The different spatial distribution of immune populations and their different association with prognosis support NAC as a modifier of tumour immune infiltrate in STS., Funding: Pharmamar; Italian Ministry of Health [RF-2019-12370923; GR-2016-02362609]; 5 × 1000 Funds-2016, Italian Ministry of Health; AIRC Grant [ID#28546]., Competing Interests: Declaration of interests Sandro Pasquali reports institutional research funds from Pharmamar, Ikena Oncology, and ASTX Pharmaceutical. Cleofe Romagosa reports institutional research funds from Pharmamar. Angelo Paolo Dei Tos reports institutional research funds from Pharmamar. Javier Martin-Broto reports institutional research funds from Pharmamar, Adaptimmune, Amgen, AROG, Bayer, Blueprint, BMS, Celgene, Daiichi Sankyo, Deciphera, Eisai, Forma, GSK, IMMIX Biopharma, Karyopharm, Lilly, LIXTE, Nektar, Novartis, Pfizer, Roche, and PharmaMar and compensations for advisory board or consulting relationship from Tecnofarma. Jean-Yves Blay reports compensations for advisory board or consulting relationship with Merck Sharp & Dohme, Merck Serono, Daiichi-Sankyo, Astellas, Alexo Oncology, Samyang Biopharm, Hanmi, Daewoong and Amgen and has received institutional research funds from Genentech/Roche, Merck Sharp & Dohme, Merck Serono, Daiichi-Sankyo, Astellas and Amgen. Giovanni Grignani reports institutional research funds from Eli Lilly and Company, GlaxoSmithKline, Merk, Novartis, and Pharmamar. Emanuela Palmerini reports compensations for advisory boards from Daiichi Sankyo Company, Daiichi Sankyo Europe GmbH, Deciphera Pharmaceuticals Inc., EUSA Pharma (US) LLC, and SynOx Therapeutics. Paolo Casali received honoraria for speaker, consultancy, or advisory roles from: Bayer, Deciphera, Eisai, Eli Lilly, and Pfizer; his unit received funds from: Advenchen Laboratories, Amgen Dompé, AROG Pharmaceuticals, Bayer, Blueprint Medicines, Daiichi Sankyo, Deciphera, Eisai, Eli Lilly, Epizyme Inc, Glaxo, Karyopharm Pharmaceuticals, Novartis, Pfizer, and PharmaMar. Silvia Stacchiotti reports institutional research funds from: Advenchen Laboratories, Amgen Dompé, AROG Pharmaceuticals, Bayer, Blueprint Medicines, Daiichi Sankyo, Deciphera, Eisai, Eli Lilly, Epizyme Inc, Glaxo Smith Kline, Karyopharm Pharmaceuticals, Novartis, Pfizer, PharmaMar, and SpringWorks; honoraria, consultancy, or advisory role from: Bayer, Bavarian Nordic, Boehringer, Deciphera, Daiichi Sankyo Pharma, Gentili, Glaxo Smith Kline, Inhibrix, Maxivax, PharmaMar, and Servier; travel, accommodations, expenses from: PharmaMar. Alessandro Gronchi reports compensations for advisory boards from Novartis, Pfizer, Bayer, Lilly, PharmaMar, SpringWorks, and Deciphera and institutional research grants from PharmaMar and Nanobiotix. The other authors declare no conflicts of interest., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)
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- 2024
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22. Multistep tumor genetic evolution and changes in immunogenicity trigger immune-mediated disease eradication in stage IV melanoma: lessons from a single case.
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Vallacchi V, Vergani E, Cossa M, Gargiuli C, Busico A, Devecchi A, Dugo M, Bergamaschi L, De Cecco L, Cavalieri S, Valeri B, Tamborini E, Gallino G, Del Vecchio M, Santinami M, Sensi M, Rivoltini L, Di Guardo L, and Rodolfo M
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- Humans, Ipilimumab therapeutic use, Vemurafenib, T-Lymphocytes pathology, Receptors, Antigen, T-Cell therapeutic use, Tumor Microenvironment, Melanoma drug therapy, Melanoma genetics, Melanoma pathology
- Abstract
Durable remissions are observed in 10%-20% of treated patients with advanced metastatic melanoma but the factors associated with long-term complete clinical responses are largely unknown. Here, we report the molecular characteristics of tumor evolution during disease progression along a 9-year clinical course in a patient with advanced disseminated melanoma who received different treatments, including trametinib, ipilimumab, radiation, vemurafenib, surgical tumor debulking and a second ipilimumab course, ultimately achieving complete long-term disease remission.Longitudinal analyses of therapies-resistant metastatic tumors revealed the effects of different treatments on tumor's microenvironment and immunogenicity, ultimately creating a milieu favorable to immunotherapy response. Monitoring of the temporal dynamics of T cells by analysis of the T cell receptor (TCR) repertoire in the tumor and peripheral blood during disease evolution indicated that T-cell clones with common TCR rearrangements, present at low levels at baseline, were maintained and expanded after immunotherapy, and that TCR diversity increased. Analysis of genetic, molecular, and cellular components of the tumor depicted a multistep process in which treatment with kinase inhibitors strongly conditioned the immune microenvironment creating an inflamed milieu converting cold into hot tumors, while ipilimumab impacted and increased the TCR repertoire, a requirement for tumor rejection.Since the optimal sequencing of treatment with antibodies targeting immune checkpoints and kinase inhibitors for advanced melanoma is still clinically debated, this case indicates that immunotherapy success is possible even after progression on targeted therapy., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)
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- 2024
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23. Development of a Molecular Blood-Based Immune Signature Classifier as Biomarker for Risks Assessment in Lung Cancer Screening.
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Fortunato O, Huber V, Segale M, Cova A, Vallacchi V, Squarcina P, Rivoltini L, Suatoni P, Sozzi G, Pastorino U, and Boeri M
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- Humans, Early Detection of Cancer methods, Leukocytes, Mononuclear, Biomarkers, Tumor genetics, Lung Neoplasms genetics, MicroRNAs genetics
- Abstract
Background: Low-dose CT (LDCT) screening trials have shown that lung cancer early detection saves lives. However, a better stratification of the screening population is still needed. In this respect, we generated and prospectively validated a plasma miRNA signature classifier (MSC) able to categorize screening participants according to lung cancer risk. Here, we aimed to deeply characterize the peripheral immune profile and develop a diagnostic immune signature classifier to further implement blood testing in lung cancer screening., Methods: Peripheral blood mononuclear cell (PBMC) samples collected from 20 patients with LDCT-detected lung cancer and 20 matched cancer-free screening volunteers were analyzed by flow cytometry using multiplex panels characterizing both lymphoid and myeloid immune subsets. Data were validated in PBMC from 40 patients with lung cancer and 40 matched controls and in a lung cancer specificity set including 27 subjects with suspicious lung nodules. A qPCR-based gene expression signature was generated resembling selected immune subsets., Results: Monocytic myeloid-derived suppressor cell (MDSC), polymorphonuclear MDSC, intermediate monocytes and CD8+PD-1+ T cells distinguished patients with lung cancer from controls with AUCs values of 0.94/0.72/0.88 in the training, validation, and lung cancer specificity set, respectively. AUCs raised up to 1.00/0.84/0.92 in subgroup analysis considering only MSC-negative subjects. A 14-immune genes expression signature distinguished patients from controls with AUC values of 0.76 in the validation set and 0.83 in MSC-negative subjects., Conclusions: An immune-based classifier can enhance the accuracy of blood testing, thus supporting the contribution of systemic immunity to lung carcinogenesis., Impact: Implementing LDCT screening trials with minimally invasive blood tests could help reduce unnecessary procedures and optimize cost-effectiveness., (©2022 The Authors; Published by the American Association for Cancer Research.)
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- 2022
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24. Genetic Layout of Melanoma Lesions Is Associated with BRAF/MEK-Targeted Therapy Resistance and Transcriptional Profiles.
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Vergani E, Busico A, Dugo M, Devecchi A, Valeri B, Cossa M, Di Guardo L, De Cecco L, Feltrin E, Valle G, Deho P, Frigerio S, Lalli L, Gallino G, Del Vecchio M, Santinami M, Pruneri G, Tamborini E, Rivoltini L, Sensi M, Vallacchi V, and Rodolfo M
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- Humans, Vemurafenib therapeutic use, Antineoplastic Combined Chemotherapy Protocols therapeutic use, MAP Kinase Kinase Kinases genetics, MAP Kinase Kinase Kinases therapeutic use, Mutation, Chromatin, Mechanistic Target of Rapamycin Complex 1, Mitogen-Activated Protein Kinase Kinases, Protein Kinase Inhibitors pharmacology, Protein Kinase Inhibitors therapeutic use, Proto-Oncogene Proteins B-raf genetics, Melanoma drug therapy, Melanoma genetics, Melanoma pathology
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The genetic landscape of melanoma resistance to targeted therapy with small molecules inhibiting BRAF and MEK kinases is still largely undefined. In this study, we portrayed in detail the somatic alterations of resistant melanoma and explored the associated biological processes and their integration with transcriptional profiles. By targeted next-generation sequencing and whole-exome sequencing analyses, a list of 101 genes showing imbalance in metastatic tumors from patients with a complete/durable response or disease progression during therapy with vemurafenib or with dabrafenib and trametinib was defined. Classification of altered genes in functional categories indicated that the mutational pattern of both resistant tumors and melanoma cell lines was enriched in gene families involved in oncogenic signaling pathways and in DNA repair. Integration of genomic and transcriptomic features showed that the enrichment of mutations in gene sets associated with anabolic processes, chromatin alterations, and IFN-α response determined a significant positive modulation of the same gene signatures at the transcriptional level. In particular, MTORC1 signaling was enriched in tumors from poorly responsive patients and in resistant tumors excised from treated patients. Results indicate that genetic patterns are associated with melanoma resistance to targeted therapy and disclose the underlying key molecular pathways to define drug combinations for improved personalized therapies., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
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- 2022
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25. Extracellular vesicles in anti-tumor immunity.
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Vergani E, Daveri E, Vallacchi V, Bergamaschi L, Lalli L, Castelli C, Rodolfo M, Rivoltini L, and Huber V
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- Humans, Immunotherapy, Immunity, Cell Line, Tumor, Extracellular Vesicles metabolism
- Abstract
To what extent extracellular vesicles (EVs) can impact anti-tumor immune responses has only started to get unraveled. Their nanometer dimensions, their growing number of subtypes together with the difficulties in defining their origin hamper their investigation. The existence of tumor cell lines facilitated advance in cancer EV understanding, while capturing information about phenotypes and functions of immune cell EVs in this context is more complex. The advent of immunotherapy with immune checkpoint inhibitors has further deepened the need to dissect the impact of EVs during immune activation and response, not least to contribute unraveling and preventing the generation of resistance occurring in the majority of patients. Here we discuss the factors that influence anddrive the immune response in cancer patients in the context of cancer therapeutics and the roles or possible functions that EVs can have in this scenario. With immune cell-derived EVs as leitmotiv, we will journey from EV discovery and subtypes through physiological and pathological functions, from similarities with tumor EVs to measures to revert detrimental consequences on immune responses to cancer., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)
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- 2022
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26. 3D models for melanoma γδ T cell-based immunotherapy.
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Huber V, Vallacchi V, Daveri E, and Vergani E
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- Humans, Receptors, Antigen, T-Cell, gamma-delta, Immunotherapy, Melanoma therapy
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- 2022
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27. Fasting-Mimicking Diet Is Safe and Reshapes Metabolism and Antitumor Immunity in Patients with Cancer.
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Vernieri C, Fucà G, Ligorio F, Huber V, Vingiani A, Iannelli F, Raimondi A, Rinchai D, Frigè G, Belfiore A, Lalli L, Chiodoni C, Cancila V, Zanardi F, Ajazi A, Cortellino S, Vallacchi V, Squarcina P, Cova A, Pesce S, Frati P, Mall R, Corsetto PA, Rizzo AM, Ferraris C, Folli S, Garassino MC, Capri G, Bianchi G, Colombo MP, Minucci S, Foiani M, Longo VD, Apolone G, Torri V, Pruneri G, Bedognetti D, Rivoltini L, and de Braud F
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- Antineoplastic Agents administration & dosage, Breast Neoplasms diet therapy, Breast Neoplasms immunology, Breast Neoplasms metabolism, Colorectal Neoplasms diet therapy, Colorectal Neoplasms immunology, Colorectal Neoplasms metabolism, Female, Humans, Male, Middle Aged, Prospective Studies, Treatment Outcome, Antineoplastic Agents therapeutic use, Breast Neoplasms drug therapy, Colorectal Neoplasms drug therapy, Fasting
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In tumor-bearing mice, cyclic fasting or fasting-mimicking diets (FMD) enhance the activity of antineoplastic treatments by modulating systemic metabolism and boosting antitumor immunity. Here we conducted a clinical trial to investigate the safety and biological effects of cyclic, five-day FMD in combination with standard antitumor therapies. In 101 patients, the FMD was safe, feasible, and resulted in a consistent decrease of blood glucose and growth factor concentration, thus recapitulating metabolic changes that mediate fasting/FMD anticancer effects in preclinical experiments. Integrated transcriptomic and deep-phenotyping analyses revealed that FMD profoundly reshapes anticancer immunity by inducing the contraction of peripheral blood immunosuppressive myeloid and regulatory T-cell compartments, paralleled by enhanced intratumor Th1/cytotoxic responses and an enrichment of IFNγ and other immune signatures associated with better clinical outcomes in patients with cancer. Our findings lay the foundations for phase II/III clinical trials aimed at investigating FMD antitumor efficacy in combination with standard antineoplastic treatments. SIGNIFICANCE: Cyclic FMD is well tolerated and causes remarkable systemic metabolic changes in patients with different tumor types and treated with concomitant antitumor therapies. In addition, the FMD reshapes systemic and intratumor immunity, finally activating several antitumor immune programs. Phase II/III clinical trials are needed to investigate FMD antitumor activity/efficacy. This article is highlighted in the In This Issue feature, p. 1 ., (©2021 The Authors; Published by the American Association for Cancer Research.)
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- 2022
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28. Liquid Biopsy and Radiological Response Predict Outcomes Following Discontinuation of Targeted Therapy in Patients with BRAF Mutated Melanoma.
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Di Guardo L, Randon G, Corti F, Vallacchi V, Raimondi A, Fucà G, Bini M, Maurichi A, Patuzzo R, Gallino G, Mattavelli I, Ruggeri R, Angi M, Cossa M, Valeri B, Cimminiello C, Santinami M, Rivoltini L, de Braud F, Rodolfo M, and Vecchio MD
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- Humans, Liquid Biopsy, Proto-Oncogene Proteins B-raf genetics, Retrospective Studies, Melanoma drug therapy, Melanoma genetics, Neoplasms, Second Primary
- Abstract
Background: Outcomes of patients with metastatic melanoma discontinuing BRAF-targeted therapy for cumulative toxicity after sustained response are unknown., Materials and Methods: This retrospective case series analysis conducted at a single Cancer Center in Italy included patients with BRAF mutated metastatic melanoma treated with a BRAF inhibitor as a single agent or in combination with a MEK inhibitor between June 1, 2011 and January 1, 2020 and interrupted treatment due to cumulative toxicity after achieving complete response (CR) or long-lasting partial response (PR; i.e. >12 months)., Results: We included 24 patients with a median treatment duration of 59.4 months (95% confidence interval [CI], 55.4-63.4; range, 12-88). CR and PR were achieved in 71% and 29% of patients, respectively. At a median follow-up after treatment discontinuation of 37.8 months (95% CI, 33.7-41.9), the 12-month progression-free survival after discontinuation (dPFS) rate was 70.8% (95% CI 54.8-91.6) and 24-month dPFS rate was 58.3% (95% CI, 41.6-81.8). Baseline patient and tumor characteristics as well as treatment duration and best response did not significantly impact on dPFS. Patients with CR and negative circulating tumor DNA (ctDNA) at time of discontinuation had a significantly improved dPFS compared with patients with either radiological residual disease or ctDNA positivity (p = .007). No patient in CR with undetectable ctDNA experienced progression., Conclusion: The risk of progression is high even in patients with sustained sensitivity to BRAF/MEK inhibitors. Integration of liquid biopsy in clinical trials investigating the optimal management of patients with sustained sensitivity to BRAF/MEK inhibitors is warranted., Implications for Practice: Outcomes of patients with metastatic melanoma discontinuing BRAF-targeted therapy for cumulative toxicity are unknown. This study analyzed patients with sustained responses (median treatment duration 59.4 months). Twelve- and 24-month progression-free survival following discontinuation were 70.8% and 58.3%, respectively. Complete response and negative circulating tumor DNA at time of discontinuation are promising prognostic biomarkers in this setting., (© 2021 The Authors. The Oncologist published by Wiley Periodicals LLC on behalf of AlphaMed Press.)
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- 2021
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29. Genetic Variants and Somatic Alterations Associated with MITF-E318K Germline Mutation in Melanoma Patients.
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Vergani E, Frigerio S, Dugo M, Devecchi A, Feltrin E, De Cecco L, Vallacchi V, Cossa M, Di Guardo L, Manoukian S, Peissel B, Ferrari A, Gallino G, Maurichi A, Rivoltini L, Sensi M, and Rodolfo M
- Subjects
- Adult, Aged, Aged, 80 and over, Chromosomes, Human, Pair 9, Cyclin-Dependent Kinase Inhibitor p16 genetics, Female, Heterozygote, Humans, Male, Middle Aged, Proto-Oncogene Proteins B-raf genetics, Receptor, Melanocortin, Type 1 genetics, Exome Sequencing, Young Adult, Melanoma, Cutaneous Malignant, Germ-Line Mutation, Melanoma genetics, Microphthalmia-Associated Transcription Factor genetics, Skin Neoplasms genetics
- Abstract
The MITF-E318K variant has been implicated in genetic predisposition to cutaneous melanoma. We addressed the occurrence of MITF-E318K and its association with germline status of CDKN2A and MC1R genes in a hospital-based series of 248 melanoma patients including cohorts of multiple, familial, pediatric, sporadic and melanoma associated with other tumors. Seven MITF-E318K carriers were identified, spanning every group except the pediatric patients. Three carriers showed mutated CDKN2A, five displayed MC1R variants, while the sporadic carrier revealed no variants. Germline/tumor whole exome sequencing for this carrier revealed germline variants of unknown significance in ATM and FANCI genes and, in four BRAF-V600E metastases, somatic loss of the MITF wild-type allele, amplification of MITF-E318K and deletion of a 9p21.3 chromosomal region including CDKN2A and MTAP. In silico analysis of tumors from MITF-E318K melanoma carriers in the TCGA Pan-Cancer-Atlas dataset confirmed the association with BRAF mutation and 9p21.3 deletion revealing a common genetic pattern. MTAP was the gene deleted at homozygous level in the highest number of patients. These results support the utility of both germline and tumor genome analysis to define tumor groups providing enhanced information for clinical strategies and highlight the importance of melanoma prevention programs for MITF-E318K patients.
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- 2021
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30. Molecular Determinants of Soft Tissue Sarcoma Immunity: Targets for Immune Intervention.
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Tazzari M, Bergamaschi L, De Vita A, Collini P, Barisella M, Bertolotti A, Ibrahim T, Pasquali S, Castelli C, and Vallacchi V
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- Animals, Biomarkers, Tumor immunology, Humans, Sarcoma drug therapy, Sarcoma pathology, Biomarkers, Tumor antagonists & inhibitors, Gene Expression Regulation, Neoplastic, Immunotherapy methods, Molecular Targeted Therapy, Sarcoma immunology, Tumor Microenvironment immunology
- Abstract
Soft tissue sarcomas (STSs) are a family of rare malignant tumors encompassing more than 80 histologies. Current therapies for metastatic STS, a condition that affects roughly half of patients, have limited efficacy, making innovative therapeutic strategies urgently needed. From a molecular point of view, STSs can be classified as translocation-related and those with a heavily rearranged genotype. Although only the latter display an increased mutational burden, molecular profiles suggestive of an "immune hot" tumor microenvironment are observed across STS histologies, and response to immunotherapy has been reported in both translocation-related and genetic complex STSs. These data reinforce the notion that immunity in STSs is multifaceted and influenced by both genetic and epigenetic determinants. Cumulative evidence indicates that a fine characterization of STSs at different levels is required to identify biomarkers predictive of immunotherapy response and to discover targetable pathways to switch on the immune sensitivity of "immune cold" tumors. In this review, we will summarize recent findings on the interplay between genetic landscape, molecular profiling and immunity in STSs. Immunological and molecular features will be discussed for their prognostic value in selected STS histologies. Finally, the local and systemic immunomodulatory effects of the targeted drugs imatinib and sunitinib will be discussed.
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- 2021
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31. Integrated transcriptional-phenotypic analysis captures systemic immunomodulation following antiangiogenic therapy in renal cell carcinoma patients.
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Rinchai D, Verzoni E, Huber V, Cova A, Squarcina P, De Cecco L, de Braud F, Ratta R, Dugo M, Lalli L, Vallacchi V, Rodolfo M, Roelands J, Castelli C, Chaussabel D, Procopio G, Bedognetti D, and Rivoltini L
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- Aged, Carcinoma, Renal Cell drug therapy, Carcinoma, Renal Cell metabolism, Carcinoma, Renal Cell pathology, Female, Follow-Up Studies, Gene Expression Regulation, Neoplastic, Humans, Kidney Neoplasms drug therapy, Kidney Neoplasms metabolism, Kidney Neoplasms pathology, Male, Myeloid-Derived Suppressor Cells immunology, Prognosis, Survival Rate, Transcriptome, Tumor Cells, Cultured, Angiogenesis Inhibitors therapeutic use, Biomarkers, Tumor genetics, Carcinoma, Renal Cell immunology, Immunomodulation, Indazoles therapeutic use, Kidney Neoplasms immunology, Pyrimidines therapeutic use, Sulfonamides therapeutic use, Tumor Microenvironment
- Abstract
Background: The combination of immune checkpoint blockade (ICB) with standard therapies is becoming a common approach for overcoming resistance to cancer immunotherapy in most human malignancies including metastatic renal cell carcinoma (mRCC). In this regard, insights into the immunomodulatory properties of antiangiogenic agents may help designing multidrug schedules based on specific immune synergisms., Methods: We used orthogonal transcriptomic and phenotyping platforms combined with functional analytic pipelines to elucidate the immunomodulatory effect of the antiangiogenic agent pazopanib in mRCC patients. Nine patients were studied longitudinally over a period of 6 months. We also analyzed transcriptional data from The Cancer Genome Atlas (TCGA) RCC cohort (N = 571) to assess the prognostic implications of our findings. The effect of pazopanib was assessed in vitro on NK cells and T cells. Additionally, myeloid-derived suppressor (MDSC)-like cells were generated from CD14
+ monocytes transfected with mimics of miRNAs associated with MDSC function in the presence or absence of pazopanib., Results: Pazopanib administration caused a rapid and dramatic reshaping in terms of frequency and transcriptional activity of multiple blood immune cell subsets, with a downsizing of MDSC and regulatory T cells in favor of a strong enhancement in PD-1 expressing cytotoxic T and Natural Killer effectors. These changes were paired with an increase of the expression of transcripts reflecting activation of immune-effector functions. This immunomodulation was marked but transient, peaking at the third month of treatment. Moreover, the intratumoral expression level of a MDSC signature (MDSC INT) was strongly associated with poor prognosis in RCC patients. In vitro experiments indicate that the observed immunomodulation might be due to an inhibitory effect on MDSC-mediated suppression, rather than a direct effect on NK and T cells., Conclusions: The marked but transient nature of this immunomodulation, peaking at the third month of treatment, provides the rationale for the use of antiangiogenics as a preconditioning strategy to improve the efficacy of ICB., (© 2021 The Authors. Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.)- Published
- 2021
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32. miR-146a-5p impairs melanoma resistance to kinase inhibitors by targeting COX2 and regulating NFkB-mediated inflammatory mediators.
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Vergani E, Dugo M, Cossa M, Frigerio S, Di Guardo L, Gallino G, Mattavelli I, Vergani B, Lalli L, Tamborini E, Valeri B, Gargiuli C, Shahaj E, Ferrarini M, Ferrero E, Gomez Lira M, Huber V, Del Vecchio M, Sensi M, Leone BE, Santinami M, Rivoltini L, Rodolfo M, and Vallacchi V
- Subjects
- Cell Line, Tumor, Cyclooxygenase 2 genetics, Extracellular Signal-Regulated MAP Kinases metabolism, Gene Expression Regulation, Neoplastic drug effects, Humans, Melanoma pathology, MicroRNAs genetics, Mitogen-Activated Protein Kinase Kinases antagonists & inhibitors, Mitogen-Activated Protein Kinase Kinases metabolism, Models, Biological, Protein Kinase Inhibitors pharmacology, Proto-Oncogene Proteins B-raf antagonists & inhibitors, Proto-Oncogene Proteins B-raf metabolism, Proto-Oncogene Proteins c-akt metabolism, Signal Transduction drug effects, Cyclooxygenase 2 metabolism, Drug Resistance, Neoplasm drug effects, Drug Resistance, Neoplasm genetics, Inflammation Mediators metabolism, Melanoma drug therapy, Melanoma genetics, MicroRNAs metabolism, NF-kappa B metabolism, Protein Kinase Inhibitors therapeutic use
- Abstract
Background: Targeted therapy with BRAF and MEK inhibitors has improved the survival of patients with BRAF-mutated metastatic melanoma, but most patients relapse upon the onset of drug resistance induced by mechanisms including genetic and epigenetic events. Among the epigenetic alterations, microRNA perturbation is associated with the development of kinase inhibitor resistance. Here, we identified and studied the role of miR-146a-5p dysregulation in melanoma drug resistance., Methods: The miR-146a-5p-regulated NFkB signaling network was identified in drug-resistant cell lines and melanoma tumor samples by expression profiling and knock-in and knock-out studies. A bioinformatic data analysis identified COX2 as a central gene regulated by miR-146a-5p and NFkB. The effects of miR-146a-5p/COX2 manipulation were studied in vitro in cell lines and with 3D cultures of treatment-resistant tumor explants from patients progressing during therapy., Results: miR-146a-5p expression was inversely correlated with drug sensitivity and COX2 expression and was reduced in BRAF and MEK inhibitor-resistant melanoma cells and tissues. Forced miR-146a-5p expression reduced COX2 activity and significantly increased drug sensitivity by hampering prosurvival NFkB signaling, leading to reduced proliferation and enhanced apoptosis. Similar effects were obtained by inhibiting COX2 by celecoxib, a clinically approved COX2 inhibitor., Conclusions: Deregulation of the miR-146a-5p/COX2 axis occurs in the development of melanoma resistance to targeted drugs in melanoma patients. This finding reveals novel targets for more effective combination treatment. Video Abstract.
- Published
- 2020
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33. microRNAs Shape Myeloid Cell-Mediated Resistance to Cancer Immunotherapy.
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Daveri E, Vergani E, Shahaj E, Bergamaschi L, La Magra S, Dosi M, Castelli C, Rodolfo M, Rivoltini L, Vallacchi V, and Huber V
- Subjects
- Drug Resistance, Neoplasm genetics, Epigenesis, Genetic, Humans, Immunomodulation genetics, Immunosuppression Therapy, Immunotherapy, Myeloid-Derived Suppressor Cells immunology, Myeloid-Derived Suppressor Cells metabolism, Neoplasms metabolism, Neoplasms therapy, Tumor Escape immunology, MicroRNAs genetics, Myeloid Cells immunology, Myeloid Cells metabolism, Neoplasms genetics, Neoplasms immunology
- Abstract
Immunotherapy with immune checkpoint inhibitors can achieve long-term tumor control in subsets of patients. However, its effect can be blunted by myeloid-induced resistance mechanisms. Myeloid cells are highly plastic and physiologically devoted to wound healing and to immune homeostasis maintenance. In cancer, their physiological activities can be modulated, leading to an expansion of pro-inflammatory and immunosuppressive cells, the myeloid-derived suppressor cells (MDSCs), with detrimental consequences. The involvement of MDSCs in tumor development and progression has been widely investigated and MDSC-induced immunosuppression is acknowledged as a mechanism hindering effective immune checkpoint blockade. Small non-coding RNA molecules, the microRNAs (miRs), contribute to myeloid cell regulation at different levels, comprising metabolism and function, as well as their skewing to a MDSC phenotype. miR expression can be indirectly induced by cancer-derived factors or through direct miR import via extracellular vesicles. Due to their structural stability and their presence in body fluids miRs represent promising predictive biomarkers of resistance, as we recently found by investigating plasma samples of melanoma patients undergoing immune checkpoint blockade. Dissection of the miR-driven involved mechanisms would pave the way for the identification of new druggable targets. Here, we discuss the role of these miRs in shaping myeloid resistance to immunotherapy with a special focus on immunosuppression and immune escape., (Copyright © 2020 Daveri, Vergani, Shahaj, Bergamaschi, La Magra, Dosi, Castelli, Rodolfo, Rivoltini, Vallacchi and Huber.)
- Published
- 2020
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34. Network modeling of patients' biomolecular profiles for clinical phenotype/outcome prediction.
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Gliozzo J, Perlasca P, Mesiti M, Casiraghi E, Vallacchi V, Vergani E, Frasca M, Grossi G, Petrini A, Re M, Paccanaro A, and Valentini G
- Subjects
- Algorithms, Artificial Intelligence, Breast Neoplasms epidemiology, Colorectal Neoplasms epidemiology, Computational Biology methods, Datasets as Topic, Female, Humans, Individuality, Male, Pancreatic Neoplasms epidemiology, Phenotype, Prognosis, Transcriptome, Treatment Outcome, Breast Neoplasms diagnosis, Colorectal Neoplasms diagnosis, Gene Regulatory Networks, Neural Networks, Computer, Pancreatic Neoplasms diagnosis
- Abstract
Methods for phenotype and outcome prediction are largely based on inductive supervised models that use selected biomarkers to make predictions, without explicitly considering the functional relationships between individuals. We introduce a novel network-based approach named Patient-Net (P-Net) in which biomolecular profiles of patients are modeled in a graph-structured space that represents gene expression relationships between patients. Then a kernel-based semi-supervised transductive algorithm is applied to the graph to explore the overall topology of the graph and to predict the phenotype/clinical outcome of patients. Experimental tests involving several publicly available datasets of patients afflicted with pancreatic, breast, colon and colorectal cancer show that our proposed method is competitive with state-of-the-art supervised and semi-supervised predictive systems. Importantly, P-Net also provides interpretable models that can be easily visualized to gain clues about the relationships between patients, and to formulate hypotheses about their stratification.
- Published
- 2020
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35. Pazopanib for treatment of advanced extraskeletal myxoid chondrosarcoma: a multicentre, single-arm, phase 2 trial.
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Stacchiotti S, Ferrari S, Redondo A, Hindi N, Palmerini E, Vaz Salgado MA, Frezza AM, Casali PG, Gutierrez A, Lopez-Pousa A, Grignani G, Italiano A, LeCesne A, Dumont S, Blay JY, Penel N, Bernabeu D, de Alava E, Karanian M, Morosi C, Brich S, Dagrada GP, Vallacchi V, Castelli C, Brenca M, Racanelli D, Maestro R, Collini P, Cruz J, and Martin-Broto J
- Subjects
- Aged, Antineoplastic Combined Chemotherapy Protocols adverse effects, Chondrosarcoma pathology, Disease-Free Survival, Female, Humans, Indazoles, Male, Middle Aged, Neoplasm Staging, Neoplasms, Connective and Soft Tissue pathology, Pyrimidines adverse effects, Retrospective Studies, Soft Tissue Neoplasms pathology, Sulfonamides adverse effects, Treatment Outcome, Antineoplastic Combined Chemotherapy Protocols administration & dosage, Chondrosarcoma drug therapy, Neoplasms, Connective and Soft Tissue drug therapy, Pyrimidines administration & dosage, Soft Tissue Neoplasms drug therapy, Sulfonamides administration & dosage
- Abstract
Background: Extraskeletal myxoid chondrosarcoma is a rare sarcoma with low sensitivity to cytotoxic chemotherapy. Retrospective evidence suggests that antiangiogenic drugs could be a treatment option. We aimed to investigate the activity of pazopanib, an antiangiogenic drug, in patients with advanced extraskeletal myxoid chondrosarcoma., Methods: In this single-arm, open-label phase 2 trial, three parallel independent cohorts of different histotypes of advanced sarcomas were recruited (extraskeletal myxoid chondrosarcoma, typical solitary fibrous tumour, and malignant-dedifferentiated solitary fibrous tumour). In each cohort, patients received pazopanib. In this Article, we report the results of the cohort of patients with advanced extraskeletal myxoid chondrosarcoma. Separate reporting of the three cohorts was prespecified in the study protocol. In this cohort, adult patients (aged ≥18 years) with a diagnosis of NR4A3-translocated, metastatic, or unresectable extraskeletal myxoid chondrosarcoma, who had Response Evaluation Criteria in Solid Tumors (RECIST) progression in the previous 6 months, and had an Eastern Cooperative Oncology Group performance status of 0-2, were enrolled at 11 study sites of the Spanish, Italian, and French sarcoma groups. Patients received oral pazopanib (800 mg/day) continuously, until disease progression, unacceptable toxicity, death, non-compliance, patient refusal, or investigator's decision. The primary endpoint was the proportion of patients achieving an objective response according to RECIST 1·1 in the modified intention-to-treat population (patients who provided consent and had a central molecularly confirmed diagnosis of extraskeletal myxoid chondrosarcoma). The safety analysis included all patients who received at least one dose of pazopanib. This study is registered with ClinicalTrials.gov, number NCT02066285., Findings: Between June 24, 2014, and Jan 17, 2017, 26 patients entered the study and started pazopanib. Of these, 23 met the eligibility criteria for the modified intention-to-treat analysis. Median follow-up was 27 months (IQR 18-30). 22 patients (one patient died before the primary analysis) were evaluable for the primary endpoint: four (18% [95% CI 1-36]) had a RECIST objective response. No deaths or grade 4 adverse events occurred. The most frequent grade 3 adverse events were hypertension (nine [35%] of 26 patients), increased concentration of alanine aminotransferase (six [23%]), and increased aspartate aminotransferase (five [19%])., Interpretation: Pazopanib had clinically meaningful antitumour activity in patients with progressive and advanced extraskeletal myxoid chondrosarcoma, and could be considered a suitable option after failure to respond to first-line anthracycline-based chemotherapy in these patients., Funding: Spanish Group for Research on Sarcomas, Italian Sarcoma Group, French Sarcoma Group, GlaxoSmithKline, and Novartis., (Copyright © 2019 Elsevier Ltd. All rights reserved.)
- Published
- 2019
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36. Immunosuppressive circuits in tumor microenvironment and their influence on cancer treatment efficacy.
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Tuccitto A, Shahaj E, Vergani E, Ferro S, Huber V, Rodolfo M, Castelli C, Rivoltini L, and Vallacchi V
- Subjects
- Animals, Humans, Immunotherapy methods, Neoplasms immunology, Tumor Escape immunology, Tumor Microenvironment immunology
- Abstract
It has been for long conceived that hallmarks of cancer were intrinsic genetic features driving tumor development, proliferation, and progression, and that targeting such cell-autonomous pathways could be sufficient to achieve therapeutic cancer control. Clinical ex vivo data demonstrated that treatment efficacy often relied on the contribution of host immune responses, hence introducing the concept of tumor microenvironment (TME), namely the existence, along with tumor cells, of non-tumor components that could significantly influence tumor growth and survival. Among the complex network of TME-driving forces, immunity plays a key role and the balance between antitumor and protumor immune responses is a major driver in contrasting or promoting cancer spreading. TME is usually a very immunosuppressed milieu because of a vast array of local alterations contrasting antitumor adaptive immunity, where metabolic changes contribute to cancer dissemination by impairing T cell infiltration and favoring the accrual and activation of regulatory cells. Subcellular structures known as extracellular vesicles then help spreading immunosuppression at systemic levels by distributing genetic and protein tumor repertoire in distant tissues. A major improvement in the knowledge of TME is now pointing the attention back to tumor cells; indeed, recent findings are showing how oncogenic pathways and specific mutations in tumor cells can actually dictate the nature and the function of immune infiltrate. As our information on the reciprocal interactions regulating TME increases, finding a strategy to interfere with TME crosstalk becomes more complex and challenging. Nevertheless, TME interactions represent a promising field for the discovery of novel biomarkers and therapeutic targets for improving treatment efficacy in cancer.
- Published
- 2019
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37. Tumor-derived microRNAs induce myeloid suppressor cells and predict immunotherapy resistance in melanoma.
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Huber V, Vallacchi V, Fleming V, Hu X, Cova A, Dugo M, Shahaj E, Sulsenti R, Vergani E, Filipazzi P, De Laurentiis A, Lalli L, Di Guardo L, Patuzzo R, Vergani B, Casiraghi E, Cossa M, Gualeni A, Bollati V, Arienti F, De Braud F, Mariani L, Villa A, Altevogt P, Umansky V, Rodolfo M, and Rivoltini L
- Subjects
- Animals, Female, Humans, Leukocytes, Mononuclear pathology, Male, Melanoma, Experimental pathology, Melanoma, Experimental therapy, Mice, Myeloid-Derived Suppressor Cells pathology, Immunotherapy, Leukocytes, Mononuclear immunology, Melanoma, Experimental immunology, MicroRNAs metabolism, Myeloid-Derived Suppressor Cells immunology, RNA, Neoplasm immunology
- Abstract
The accrual of myeloid-derived suppressor cells (MDSCs) represents a major obstacle to effective immunotherapy in cancer patients, but the mechanisms underlying this process in the human setting remain elusive. Here, we describe a set of microRNAs (miR-146a, miR-155, miR-125b, miR-100, let-7e, miR-125a, miR-146b, miR-99b) that are associated with MDSCs and resistance to treatment with immune checkpoint inhibitors in melanoma patients. The miRs were identified by transcriptional analyses as being responsible for the conversion of monocytes into MDSCs (CD14+HLA-DRneg cells) mediated by melanoma extracellular vesicles (EVs) and were shown to recreate MDSC features upon transfection. In melanoma patients, these miRs were increased in circulating CD14+ monocytes, plasma, and tumor samples, where they correlated with the myeloid cell infiltrate. In plasma, their baseline levels clustered with the clinical efficacy of CTLA-4 or programmed cell death protein 1 (PD-1) blockade. Hence, MDSC-related miRs represent an indicator of MDSC activity in cancer patients and a potential blood marker of a poor immunotherapy outcome.
- Published
- 2018
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38. microRNA Expression in Sentinel Nodes from Progressing Melanoma Patients Identifies Networks Associated with Dysfunctional Immune Response.
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Vallacchi V, Camisaschi C, Dugo M, Vergani E, Deho P, Gualeni A, Huber V, Gloghini A, Maurichi A, Santinami M, Sensi M, Castelli C, Rivoltini L, and Rodolfo M
- Abstract
Sentinel node biopsy (SNB) is a main staging biomarker in melanoma and is the first lymph node to drain the tumor, thus representing the immunological site where anti-tumor immune dysfunction is established and where potential prognostic immune markers can be identified. Here we analyzed microRNA (miR) profiles in archival tumor-positive SNBs derived from melanoma patients with different outcomes and performed an integrated analysis of transcriptional data to identify deregulated immune signaling networks. Twenty-six miRs were differentially expressed in melanoma-positive SNB samples between patients with disease progression and non-progressing patients, the majority being previously reported in the regulation of immune responses. A significant variation in miR expression levels was confirmed in an independent set of SNB samples. Integrated information from genome-wide transcriptional profiles and in vitro assessment in immune cells led to the identification of miRs associated with the regulation of the TNF receptor superfamily member 8 ( TNFRSF8 ) gene encoding the CD30 receptor, a marker increased in lymphocytes of melanoma patients with progressive disease. These findings indicate that miRs are involved in the regulation of pathways leading to immune dysfunction in the sentinel node and may provide valuable markers for developing prognostic molecular signatures for the identification of stage III melanoma patients at risk of recurrence., Competing Interests: The authors declare no conflict of interest.
- Published
- 2016
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39. Targeting Immune Regulatory Networks to Counteract Immune Suppression in Cancer.
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Camisaschi C, Vallacchi V, Vergani E, Tazzari M, Ferro S, Tuccitto A, Kuchuk O, Shahaj E, Sulsenti R, Castelli C, Rodolfo M, Rivoltini L, and Huber V
- Abstract
The onset of cancer is unavoidably accompanied by suppression of antitumor immunity. This occurs through mechanisms ranging from the progressive accumulation of regulatory immune cells associated with chronic immune stimulation and inflammation, to the expression of immunosuppressive molecules. Some of them are being successfully exploited as therapeutic targets, with impressive clinical results achieved in patients, as in the case of immune checkpoint inhibitors. To limit immune attack, tumor cells exploit specific pathways to render the tumor microenvironment hostile for antitumor effector cells. Local acidification might, in fact, anergize activated T cells and facilitate the accumulation of immune suppressive cells. Moreover, the release of extracellular vesicles by tumor cells can condition distant immune sites contributing to the onset of systemic immune suppression. Understanding which mechanisms may be prevalent in specific cancers or disease stages, and identifying possible strategies to counterbalance would majorly contribute to improving clinical efficacy of cancer immunotherapy. Here, we intend to highlight these mechanisms, how they could be targeted and the tools that might be available in the near future to achieve this goal., Competing Interests: The authors declare no conflict of interest.
- Published
- 2016
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40. Overcoming melanoma resistance to vemurafenib by targeting CCL2-induced miR-34a, miR-100 and miR-125b.
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Vergani E, Di Guardo L, Dugo M, Rigoletto S, Tragni G, Ruggeri R, Perrone F, Tamborini E, Gloghini A, Arienti F, Vergani B, Deho P, De Cecco L, Vallacchi V, Frati P, Shahaj E, Villa A, Santinami M, De Braud F, Rivoltini L, and Rodolfo M
- Subjects
- Adult, Aged, Blotting, Western, Case-Control Studies, Chemokine CCL2 genetics, Female, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, Humans, Male, Melanoma drug therapy, Melanoma metabolism, Melanoma pathology, Middle Aged, Neoplasm Staging, Prognosis, RNA, Messenger genetics, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Survival Rate, Tumor Cells, Cultured, Vemurafenib, Chemokine CCL2 metabolism, Drug Resistance, Neoplasm genetics, Indoles pharmacology, Melanoma genetics, MicroRNAs genetics, Sulfonamides pharmacology
- Abstract
In melanoma, the adaptative cell response to BRAF inhibitors includes altered patterns of cytokine production contributing to tumor progression and drug resistance. Among the factors produced by PLX4032-resistant melanoma cell lines, CCL2 was higher compared to the sensitive parental cell lines and increased upon drug treatment. CCL2 acted as an autocrine growth factor for melanoma cells, stimulating the proliferation and resistance to apoptosis. In patients, CCL2 is detected in melanoma cells in tumors and in plasma at levels that correlate with tumor burden and lactate dehydrogenase. Vemurafenib treatment increased the CCL2 levels in plasma, whereas the long-term clinical response was associated with low CCL2 levels.Increased CCL2 production was associated with miRNA deregulation in the resistant cells. miR-34a, miR-100 and miR-125b showed high expression in both resistant cells and in tumor biopsies that were obtained from treated patients, and they were involved in the control of cell proliferation and apoptosis. Inhibition of CCL2 and of the selected miRNAs restored both the cell apoptosis and the drug efficacy in resistant melanoma cells. Therefore, CCL2 and miRNAs are potential prognostic factors and attractive targets for counteracting treatment resistance in metastatic melanoma.
- Published
- 2016
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41. Immune cells in the melanoma microenvironment hold information for prediction of the risk of recurrence and response to treatment.
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Camisaschi C, Vallacchi V, Castelli C, Rivoltini L, and Rodolfo M
- Subjects
- Gene Expression Regulation, Neoplastic, Humans, Immunotherapy, Lymphocytes, Tumor-Infiltrating pathology, Melanoma genetics, Melanoma therapy, Neoplasm Metastasis, Neoplasm Recurrence, Local, Prognosis, Treatment Outcome, Tumor Microenvironment genetics, Lymphocytes, Tumor-Infiltrating immunology, Melanoma diagnosis, Melanoma immunology, Tumor Microenvironment immunology
- Abstract
Melanoma is an immunogenic tumor and immunotherapy treatment has established an increase in disease-free and overall survival in melanoma patients. However, a complex network of immunosuppressive mechanisms has been demonstrated to occur at the tumor site and in locoregional immune districts, such as sentinel lymph nodes (SLNs). The interplay between tumor cells and the local microenvironment leads to a tumor-driven shaping of the immune response that results in a heterogeneous cellular and molecular composition of tumor infiltrating lymphocytes (TILs). Several studies have reported the potential prognostic value of TILs infiltrating primary tumors and the association of 'immune signature' in SLNs and in melanoma metastases with prognosis and responsiveness to immunotherapeutic approaches. However, a systematic and deeper characterization of the local immunological status of TILs and SLNs is still required to refine melanoma stage classification.
- Published
- 2014
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42. Transcriptional profiling of melanoma sentinel nodes identify patients with poor outcome and reveal an association of CD30(+) T lymphocytes with progression.
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Vallacchi V, Vergani E, Camisaschi C, Deho P, Cabras AD, Sensi M, De Cecco L, Bassani N, Ambrogi F, Carbone A, Crippa F, Vergani B, Frati P, Arienti F, Patuzzo R, Villa A, Biganzoli E, Canevari S, Santinami M, Castelli C, Rivoltini L, and Rodolfo M
- Subjects
- Computational Biology, Disease Progression, Humans, Immunohistochemistry, Melanoma pathology, T-Lymphocytes pathology, Transcriptome, Treatment Outcome, Ki-1 Antigen immunology, Melanoma genetics, Melanoma immunology, Sentinel Lymph Node Biopsy methods, T-Lymphocytes immunology
- Abstract
Sentinel lymph nodes set the stance of the immune system to a localized tumor and are often the first site to be colonized by neoplastic cells that metastasize. To investigate how the presence of neoplastic cells in sentinel lymph nodes may trigger pathways associated with metastatic progression, we analyzed the transcriptional profiles of archival sentinel node biopsy specimens obtained from melanoma patients. Biopsies from positive nodes were selected for comparable tumor infiltration, presence or absence of further regional node metastases, and relapse at 5-year follow-up. Unsupervised analysis of gene expression profiles revealed immune response to be a major gene ontogeny represented. Among genes upregulated in patients with progressing disease, the TNF receptor family member CD30/TNFRSF8 was confirmed in biopsy specimens from an independent group of patients. Immunohistochemical analysis revealed higher numbers of CD30(+) lymphocytes in nodes from progressing patients compared with nonprogressing patients. Phenotypic profiling demonstrated that CD30(+) lymphocytes comprised a broad population of suppressive or exhausted immune cells, such as CD4(+)Foxp3(+) or PD1(+) subpopulations and CD4(-)CD8(-) T cells. CD30(+) T lymphocytes were increased in peripheral blood lymphocytes of melanoma patients at advanced disease stages. Our findings reinforce the concept that sentinel nodes act as pivotal sites for determining progression patterns, revealing that the presence of CD30(+) lymphocytes at those sites associate positively with melanoma progression.
- Published
- 2014
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43. Identification of MET and SRC activation in melanoma cell lines showing primary resistance to PLX4032.
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Vergani E, Vallacchi V, Frigerio S, Deho P, Mondellini P, Perego P, Cassinelli G, Lanzi C, Testi MA, Rivoltini L, Bongarzone I, and Rodolfo M
- Subjects
- Cell Line, Tumor, Cell Proliferation drug effects, Drug Resistance, Neoplasm genetics, Humans, Melanoma genetics, Mutation, PTEN Phosphohydrolase genetics, PTEN Phosphohydrolase metabolism, Proto-Oncogene Proteins B-raf genetics, Proto-Oncogene Proteins c-met antagonists & inhibitors, Signal Transduction drug effects, Vemurafenib, src-Family Kinases antagonists & inhibitors, Antineoplastic Agents pharmacology, Indoles pharmacology, Melanoma enzymology, Protein Kinase Inhibitors pharmacology, Proto-Oncogene Proteins c-met metabolism, Sulfonamides pharmacology, src-Family Kinases metabolism
- Abstract
PLX4032/vemurafenib is a first-in-class small-molecule BRAF(V600E) inhibitor with clinical activity in patients with BRAF mutant melanoma. Nevertheless, drug resistance develops in treated patients, and strategies to overcome primary and acquired resistance are required. To explore the molecular mechanisms involved in primary resistance to PLX4032, we investigated its effects on cell proliferation and signaling in a panel of 27 genetically characterized patient-derived melanoma cell lines. Cell sensitivity to PLX4032 was dependent on BRAF(V600E) and independent from other gene alterations that commonly occur in melanoma such as PTEN loss, BRAF, and MITF gene amplification. Two cell lines lacking sensitivity to PLX4032 and harboring a different set of genetic alterations were studied as models of primary resistance. Treatment with the MEK inhibitor UO126 but not with PLX4032 inhibited cell growth and ERK activation. Resistance to PLX4032 was maintained after CRAF down-regulation by siRNA indicating alternative activation of MEK-ERK signaling. Genetic characterization by multiplex ligation-dependent probe amplification and analysis of phosphotyrosine signaling by MALDI-TOF mass spectrometry analysis revealed the activation of MET and SRC signaling, associated with the amplification of MET and of CTNNB1 and CCND1 genes, respectively. The combination of PLX4032 with drugs or siRNA targeting MET was effective in inhibiting cell growth and reducing cell invasion and migration in melanoma cells with MET amplification; similar effects were observed after targeting SRC in the other cell line, indicating a role for MET and SRC signaling in primary resistance to PLX4032. Our results support the development of classification of melanoma in molecular subtypes for more effective therapies.
- Published
- 2011
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44. Molecular profiling of the "plexinome" in melanoma and pancreatic cancer.
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Balakrishnan A, Penachioni JY, Lamba S, Bleeker FE, Zanon C, Rodolfo M, Vallacchi V, Scarpa A, Felicioni L, Buck M, Marchetti A, Comoglio PM, Bardelli A, and Tamagnone L
- Subjects
- Humans, Mutation, Phylogeny, Polymerase Chain Reaction, Adenocarcinoma genetics, Gene Expression Profiling, Melanoma genetics, Pancreatic Neoplasms genetics, Receptors, Cell Surface genetics
- Abstract
Plexins are transmembrane high-affinity receptors for semaphorins, regulating cell guidance, motility, and invasion. Functional evidences implicate semaphorin signals in cancer progression and metastasis. Yet, it is largely unknown whether plexin genes are genetically altered in human tumors. We performed a comprehensive gene copy analysis and mutational profiling of all nine members of the plexin gene family (plexinome), in melanomas and pancreatic ductal adenocarcinomas (PDACs), which are characterized by high metastatic potential and poor prognosis. Gene copy analysis detected amplification of PLXNA4 in melanomas, whereas copy number losses of multiple plexin genes were seen in PDACs. Somatic mutations were detected in PLXNA4, PLXNB3, and PLXNC1; providing the first evidence that these plexins are mutated in human cancer. Functional assays in cellular models revealed that some of these missense mutations result in loss of plexin function. For instance, c.1613G>A, p.R538H mutation in the extracellular domain of PLXNB3 prevented binding of the ligand Sema5A. Moreover, although PLXNA4 signaling can inhibit tumor cell migration, the mutated c.5206C>T, p.H1736Y allele had lost this activity. Our study is the first systematic analysis of the "plexinome" in human tumors, and indicates that multiple mutated plexins may be involved in cancer progression.
- Published
- 2009
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45. Regulatory role of CCN3 in melanoma cell interaction with the extracellular matrix.
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Vallacchi V and Rodolfo M
- Subjects
- Animals, Cell Adhesion, Cell Movement, Cell Proliferation, Humans, Integrins metabolism, Neoplasm Metastasis, Protein Isoforms metabolism, Cell Communication, Extracellular Matrix metabolism, Melanoma metabolism, Melanoma pathology, Nephroblastoma Overexpressed Protein metabolism
- Abstract
It is increasingly clear that melanoma cells modify their environment not only through the release of growth factors (GFs) and cytokines that have autocrine or paracrine effects and strongly modulate the immune response, but also by secreting proteins that become structural or transient components of the extracellular matrix (ECM). Melanoma cell secreted proteins play a significant role in cell-ECM interactions, helping tumor cells to invade neighbouring stroma, disseminate and survive in other tissue contexts. CCN3/NOV (nephroblastoma overexpressed) is a matricellular protein that belongs to the CCN family of proteins containing six members in humans. Its structure consists of modules related to functional domains previously identified in major regulatory proteins: insulin-like growth factor-binding protein (IGFBP), von Willebrand factor type C repeats (VWC), thrombospondin type 1 repeats, and secreted regulatory factors containing cysteine knot motifs. Extensive studies have indicated that the biological properties of CCN3 are dependent upon the cellular context, and its role in melanoma seems to recapitulate cell context functions.
- Published
- 2009
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46. CCN3/nephroblastoma overexpressed matricellular protein regulates integrin expression, adhesion, and dissemination in melanoma.
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Vallacchi V, Daniotti M, Ratti F, Di Stasi D, Deho P, De Filippo A, Tragni G, Balsari A, Carbone A, Rivoltini L, Parmiani G, Lazar N, Perbal B, and Rodolfo M
- Subjects
- Animals, Cell Adhesion physiology, Cell Line, Tumor, Connective Tissue Growth Factor, Disease Progression, Female, Humans, Immediate-Early Proteins biosynthesis, Immediate-Early Proteins genetics, Immunohistochemistry, Integrins genetics, Intercellular Signaling Peptides and Proteins biosynthesis, Intercellular Signaling Peptides and Proteins genetics, Melanoma genetics, Melanoma metabolism, Mice, Mice, SCID, Nephroblastoma Overexpressed Protein, Oligonucleotide Array Sequence Analysis, Transfection, Immediate-Early Proteins physiology, Integrins biosynthesis, Intercellular Signaling Peptides and Proteins physiology, Melanoma pathology
- Abstract
CCN3/nephroblastoma overexpressed belongs to the CCN family of genes that encode secreted proteins associated with the extracellular matrix (ECM) and exert regulatory effects at the cellular level. Overexpression of CCN3 was shown in metastatic melanoma cells compared with cells of the primary tumor from the same patient. Analysis of short-term cultures from 50 primary and metastatic melanomas revealed a heterogeneous expression pattern of both the 46-kDa full-length cytoplasmic/secreted protein and the 32-kDa nuclear-truncated form. The different protein expression patterns were not associated with gene alterations or polymorphisms. Like the metastatic cells expressing high levels of the 46-kDa CCN3, cells transfected to overexpress CCN3 showed increased adhesion to ECM proteins, whereas inhibition of CCN3 expression by small interfering RNA decreased adhesion to laminin and vitronectin. CCN3 overexpression induced increased expression of laminin and vitronectin integrin receptors alpha 7 beta 1 and alpha v beta 5 by increasing their mRNA production. Moreover, CCN3 secreted by melanoma cells acted as an adhesion matrix protein for melanoma cells themselves. Analysis of CCN3 protein expression with respect to melanoma progression detected the protein in all visceral metastases tested and in most nodal metastases from relapsing patients but in only a few nodal metastases from nonrelapsing patients and cutaneous metastases. Consistently, xenotransplantation in immunodeficient mice showed a higher metastatic potential of melanoma cells overexpressing CCN3. Together, these data indicate a role for CCN3 in melanoma cell interaction with the ECM by regulating integrin expression, resulting in altered cell adhesion and leading melanoma progression to aggressive disease.
- Published
- 2008
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47. Detection of mutated BRAFV600E variant in circulating DNA of stage III-IV melanoma patients.
- Author
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Daniotti M, Vallacchi V, Rivoltini L, Patuzzo R, Santinami M, Arienti F, Cutolo G, Pierotti MA, Parmiani G, and Rodolfo M
- Subjects
- Base Sequence, DNA Primers, Humans, Melanoma genetics, Pilot Projects, DNA, Neoplasm blood, Melanoma blood, Point Mutation, Proto-Oncogene Proteins B-raf genetics
- Abstract
BRAFV600E is the most represented somatic point mutation in cutaneous melanoma, thus providing a unique molecular marker for this disease. The development of efficient methods for its detection in free circulating DNA of patients may lead to the improvement of diagnostic and prognostic tools. With this aim, we evaluated whether BRAFV600E represents a detectable marker in the plasma/serum from melanoma patients in a pilot study. Circulating cell-free DNA was extracted from the serum or plasma of 15 healthy donors and 41 melanoma patients at different clinical stages and obtained either presurgery or after surgery during follow-up. Quantitative analysis showed higher levels of circulating free DNA in patients compared to controls, with the highest levels detected in samples obtained presurgery and at stage IV. Four different PCR methods were compared for their capacity to amplify a few copies of BRAFV600E in wild-type DNA. BRAFV600E was detectable in circulating DNA of 12 patients and in none of the controls; only 1 PCR method reproducibly amplified BRAFV600E. Positive samples were obtained from 8/13 patients at stage IV and from 4/24 patients at stage III, but not in 4 patients at stage I-II; half of the positives were obtained presurgery and half at follow-up. Correspondence between circulating DNA and related tumors were examined for 20 patients, and a correlation was found for stage IV patients. In conclusion, this method can be utilized for monitoring the disease in stage IV melanoma patients but it appears unsatisfactory for the early detection of melanoma.
- Published
- 2007
- Full Text
- View/download PDF
48. DHCR24 gene expression is upregulated in melanoma metastases and associated to resistance to oxidative stress-induced apoptosis.
- Author
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Di Stasi D, Vallacchi V, Campi V, Ranzani T, Daniotti M, Chiodini E, Fiorentini S, Greeve I, Prinetti A, Rivoltini L, Pierotti MA, and Rodolfo M
- Subjects
- Androstenes pharmacology, Antineoplastic Agents pharmacology, Antineoplastic Agents, Phytogenic pharmacology, Blotting, Northern, Cholesterol metabolism, Cisplatin pharmacology, Etoposide pharmacology, Humans, Hydrogen Peroxide pharmacology, Melanoma secondary, Nerve Tissue Proteins metabolism, Oxidants pharmacology, Oxidoreductases Acting on CH-CH Group Donors metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, Skin Neoplasms pathology, Tumor Cells, Cultured, Up-Regulation, Apoptosis, Gene Expression Regulation, Neoplastic, Melanoma genetics, Nerve Tissue Proteins genetics, Oxidative Stress, Oxidoreductases Acting on CH-CH Group Donors genetics, Skin Neoplasms genetics
- Abstract
The DHCR24 gene encoding for the 3beta-hydroxysterol delta24-reductase, an oxidoreductase involved in cholesterol biosynthesis, was isolated by subtractive hybridization as highly expressed in a short-term melanoma cell line derived from a cutaneous metastases (S/M2) compared to that obtained from the autologous primary tumor (S/P). DHCR24 (alias seladin-1, diminuto/dwarf1 homolog) has been reported to act as an antiapoptotic factor in neurons. Gene expression analysis by Northern blot confirmed that DHCR24 was 5-fold upregulated in S/M2 compared to S/P cells. High levels of DHCR24 gene expression were detected in 13/25 melanoma metastases and in 1/7 primary melanomas by real-time PCR, indicating that upregulation of this gene may occur in melanoma progression. In S/M2 cells, high DHCR24 gene expression associated with resistance to apoptosis triggered by oxidative stress induced by exposure to hydrogen peroxide. DHCR24 gene transfer was shown to protect melanoma cells from H2O2-induced cytotoxicity. Although higher cholesterol levels were shown in S/M2 cells compared to S/P cells, DHCR24 gene transfer did not increase cholesterol content. To evaluate whether DHCR24 acts as an antiapoptotic factor in melanoma metastases, the cytotoxic effect of chemotherapeutic agents was tested in DHCR24 transfectants and in the presence of a DHCR24 inhibitor, U18666A. High DHCR24 gene expression in transfectants did not result in a higher resistance to cytotoxic agents; treatment with U18666A was cytotoxic in S/P cells with a lower DHCR24 content and showed additive cytotoxic effect only when associated with H2O2 and not with cysplatin or etoposide, indicating that the DHCR24 protective effect is exerted through an oxidative stress-specific mechanism., (Copyright 2005 Wiley-Liss, Inc)
- Published
- 2005
- Full Text
- View/download PDF
49. Genetic progression of metastatic melanoma.
- Author
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Rodolfo M, Daniotti M, and Vallacchi V
- Subjects
- Disease Progression, Genetic Predisposition to Disease, Humans, Prognosis, Signal Transduction, Genetic Markers, Melanoma genetics, Melanoma physiopathology, Neoplasm Metastasis genetics, Skin Neoplasms genetics, Skin Neoplasms physiopathology
- Abstract
Melanoma progression is well defined in its clinical, histopathological and biological aspects, but the molecular mechanism involved and the genetic markers associated to metastatic dissemination are only beginning to be defined. The recent development of high-throughput technologies aimed at global molecular profiling of cancer is switching on the spotlight at previously unknown candidate genes involved in melanoma, such as WNT5A and BRAF. In fact, several tumor suppressors and oncogenes have been shown to be involved in melanoma pathogenesis, including CDKN2A, PTEN, TP53, RAS and MYC, though they have not been related to melanoma subtypes or validated as prognostic markers. Here, we have reviewed the published data relative to the major genes involved in melanoma pathogenesis, which may represent important markers for the identification of genetic profiles of melanoma subtypes.
- Published
- 2004
- Full Text
- View/download PDF
50. BRAF alterations are associated with complex mutational profiles in malignant melanoma.
- Author
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Daniotti M, Oggionni M, Ranzani T, Vallacchi V, Campi V, Di Stasi D, Torre GD, Perrone F, Luoni C, Suardi S, Frattini M, Pilotti S, Anichini A, Tragni G, Parmiani G, Pierotti MA, and Rodolfo M
- Subjects
- Adult, Aged, Aged, 80 and over, Cyclin-Dependent Kinase 4, Cyclin-Dependent Kinases genetics, Female, Genes, p16, Genes, p53, Humans, Male, Melanoma etiology, Melanoma mortality, Middle Aged, PTEN Phosphohydrolase, Phosphoric Monoester Hydrolases genetics, Promoter Regions, Genetic, Proto-Oncogene Proteins B-raf, Tumor Suppressor Proteins genetics, Melanoma genetics, Mutation, Proto-Oncogene Proteins, Proto-Oncogene Proteins c-raf genetics
- Abstract
To evaluate the mutational profiles associated with BRAF mutations in human melanoma, we have studied BRAF, RAS, PTEN, TP53, CDKN2A and CDK4 genes and their expression in melanoma lesions. Owing to the lack of sufficient material from fresh specimens, we employed short-term cell lines obtained from melanoma biopsies. In all, 41 melanoma obtained from eight primary lesions, 20 nodal, 11 cutaneous and two visceral metastases from patients with sporadic (n=31), familial (n=4) and multiple melanoma (n=2) were analysed. The results revealed novel missense mutations in the BRAF, PTEN, CDKN2A and CDK4 genes. Overall, activating mutations of BRAF and loss of functional p16 and ARF were detected in the majority of melanomas (29/41, 36/41 and 29/41, respectively), while PTEN alterations/loss, NRAS and TP53 mutations occurred less frequently (6/41, 6/41 and 10/41, respectively). In the resulting 12 mutational profiles, p16/ARF loss associated with mutated BRAFV599E was the most represented (n=15). In addition, TP53 and PTEN mutations were always accompanied with BRAF alterations, while PTEN loss was found in association with CDKN2A or TP53 mutations in the absence of BRAF activation. The p16/ARFDelta+BRAF/RAS profile was significantly associated with a longer survival, while complex mutational profiles were detected in highly aggressive disease and poor survival. These data support the existence of several molecularly defined melanoma groups which likely reflect different clinical/biological behaviour, thus suggesting that a more extensive molecular classification of melanoma would significantly impact its clinical management.
- Published
- 2004
- Full Text
- View/download PDF
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