23 results on '"Curtarello, M."'
Search Results
2. Allelic Imbalance Analysis in Liquid Biopsy to Monitor Locally Advanced Esophageal Cancer Patients During Treatment
- Author
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Boldrin, E., Curtarello, M., Fassan, M., Rugge, M., Realdon, S., Alfieri, R., Amadori, A., and Saggioro, D.
- Subjects
allelic imbalance ,circulating cell free DNA (cfDNA) ,esophageal adenocarcinoma (EADC) ,esophageal cancer ,esophageal squamous cell carcinoma (ESCC) ,liquid biopsy ,longitudinal studies ,loss of heterozygosis (LOH) ,Cancer Research ,Oncology ,Original Research - Abstract
Esophageal cancer (EC) is a highly aggressive tumor, and the current monitoring procedures are partially inadequate to evaluate treatment efficacy. The aim of this study was to investigate whether allelic imbalance analysis in liquid biopsy could be used as an additional tool to monitor tumor burden in EC patients. For this purpose, circulating cell-free DNA (cfDNA) from 52 patients with a locally advanced EC, which underwent neoadjuvant treatment and resection, was analyzed. Data from four representative longitudinally followed patients are also reported. Furthermore, 17 DNAs from formalin-fixed paraffin-embedded (FFPE) tumor samples were analyzed and compared to time-matched cfDNAs. To look for allelic imbalance, which is the main genetic alteration in both EC histotypes, we used a panel of five microsatellites (MSs) and three single-nucleotide polymorphisms (SNPs) near genes described as frequently altered. The Fisher exact and Mann-Whitney U tests were used to analyze categorical and continuous data, respectively. The correlation coefficient between cfDNA and FFPE-DNA was calculated with the Pearson's correlation test. We found that the selected tumor-related alterations are present in cfDNA of both adenocarcinoma (EADC) and squamous cell carcinoma (ESCC) with similar frequencies. The only exception were the MSs, one downstream and one upstream, of SMAD4 of which the loss was only observed in EADC (26 vs. 0%, P = 0.018). More interestingly, longitudinal studies disclosed that in patients with disease progression, tumor-related alterations were present in cfDNA before overt clinical or instrumental signs of relapse. In conclusion, our data indicate that the evaluation of tumor-related gene allelic imbalance in cfDNA might be a useful tool to complement the current monitoring procedures for EC patients and to guide their management.
- Published
- 2020
3. 19P PIK3CA mutations in HER2-positive early breast cancer patients enrolled in the adjuvant randomized short-HER study
- Author
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Guarneri, V., primary, Dieci, M.V., additional, Bisagni, G., additional, Brandes, A.A., additional, Frassoldati, A., additional, Cavanna, L., additional, Musolino, A., additional, Giotta, F., additional, Rimanti, A., additional, Garrone, O., additional, Bertone, E., additional, Cagossi, K., additional, Nanni, O., additional, Piacentini, F., additional, Orvieto, E., additional, Curtarello, M., additional, Chic, N., additional, D'Amico, R., additional, Prat, A., additional, and Conte, P.F., additional
- Published
- 2020
- Full Text
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4. Abstract P1-13-02: Withdrawn
- Author
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Guarneri, V, primary, Dieci, MV, additional, Bisagni, G, additional, Brandes, AA, additional, Frassoldati, A, additional, Cavanna, L, additional, Musolino, A, additional, Giotta, F, additional, Cavazzini, G, additional, Garrone, O, additional, Bertone, E, additional, Cagossi, K, additional, Nanni, O, additional, Ferro, A, additional, Donadio, M, additional, Aieta, M, additional, Zamagni, C, additional, Piacentini, F, additional, Maiorana, A, additional, Ragazzi, M, additional, Cucchi, MC, additional, Querzoli, P, additional, Orsi, N, additional, Curtarello, M, additional, Urso, L, additional, Amadori, A, additional, Orvieto, E, additional, Vicini, R, additional, Balduzzi, S, additional, D'Amico, R, additional, and Conte, P, additional
- Published
- 2018
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5. VEGF-targeted therapy stably modulates the glycolytic phenotype of tumor cells
- Author
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Curtarello, M, Zulato, E, Nardo, G, Valtorta, S, Guzzo, G, Rossi, E, Esposito, G, Msaki, A, Pastò, A, Rasola, A, Persano, L, Ciccarese, F, Bertorelle, R, Todde, S, Plebani, M, Schroer, H, Walenta, S, Mueller Klieser, W, Amadori, A, Moresco, R, Indraccolo, S, Valtorta, Silvia, TODDE, SERGIO CAMILLO, MORESCO, ROSA MARIA, Indraccolo, S., Curtarello, M, Zulato, E, Nardo, G, Valtorta, S, Guzzo, G, Rossi, E, Esposito, G, Msaki, A, Pastò, A, Rasola, A, Persano, L, Ciccarese, F, Bertorelle, R, Todde, S, Plebani, M, Schroer, H, Walenta, S, Mueller Klieser, W, Amadori, A, Moresco, R, Indraccolo, S, Valtorta, Silvia, TODDE, SERGIO CAMILLO, MORESCO, ROSA MARIA, and Indraccolo, S.
- Abstract
Anti-VEGF therapy perturbs tumor metabolism, severely impairing oxygen, glucose, and ATP levels. In this study, we investigated the effects of anti-VEGF therapy in multiple experimental tumor models that differ in their glycolytic phenotypes to gain insights into optimal modulation of the metabolic features of this therapy. Prolonged treatments induced vascular regression and necrosis in tumor xenograft models, with highly glycolytic tumors becoming treatment resistant more rapidly than poorly glycolytic tumors. By PET imaging, prolonged treatments yielded an increase in both hypoxic and proliferative regions of tumors. A selection for highly glycolytic cells was noted and this metabolic shift was stable and associated with increased tumor aggressiveness and resistance to VEGF blockade in serially transplanted mice. Our results support the hypothesis that the highly glycolytic phenotype of tumor cells studied in xenograft models, either primary or secondary, is a cell-autonomous trait conferring resistance to VEGF blockade. The finding that metabolic traits of tumors can be selected by antiangiogenic therapy suggests insights into the evolutionary dynamics of tumor metabolism.
- Published
- 2015
6. AMPK inhibition enhances apoptosis in MLL-rearranged pediatric B-acute lymphoblastic leukemia cells
- Author
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Accordi, B., Galla, L., Milani, G., Curtarello, M., Serafin, V., Lissandron, V., Viola, G., Te Kronnie, G., De Maria Marchiano, Ruggero, Petricoin, E. F., Liotta, L. A., Indraccolo, S., Basso, G., De Maria, R. (ORCID:0000-0003-2255-0583), Accordi, B., Galla, L., Milani, G., Curtarello, M., Serafin, V., Lissandron, V., Viola, G., Te Kronnie, G., De Maria Marchiano, Ruggero, Petricoin, E. F., Liotta, L. A., Indraccolo, S., Basso, G., and De Maria, R. (ORCID:0000-0003-2255-0583)
- Abstract
The serine/threonine kinase AMP-activated protein kinase (AMPK) and its downstream effectors, including endothelial nitric oxide synthase and BCL-2, are hyperactivated in B-cell precursor-acute lymphoblastic leukemia (BCP-ALL) cells with MLL gene rearrangements. We investigated the role of activated AMPK in supporting leukemic cell survival and evaluated AMPK as a potential drug target. Exposure of leukemic cells to the commercial AMPK inhibitor compound C resulted in massive apoptosis only in cells with MLL gene rearrangements. These results were confirmed by targeting AMPK with specific short hairpin RNAs. Compound C-induced apoptosis was associated with mitochondrial membrane depolarization, reactive oxygen species production, cytochrome c release and caspases cleavage, indicating intrinsic apoptosis pathway activation. Treatment with low concentrations of compound C resulted in a strong antileukemic activity, together with cytochrome c release and cleavage of caspases and poly(ADP-ribose) polymerase, also in MLL-rearranged primary BCP-ALL samples. Moreover, AMPK inhibition in MLL-rearranged cell lines synergistically enhanced the antiproliferative effects of vincristine, daunorubicin, cytarabine, dexamethasone and L-asparaginase in most of the evaluated conditions. Taken together, these results indicate that the activation of the AMPK pathway directly contributes to the survival of MLL-rearranged BCP-ALL cells and AMPK inhibitors could represent a new therapeutic strategy for this high-risk leukemia. © 2013 Macmillan Publishers Limited All rights reserved.
- Published
- 2013
7. AMPK inhibition enhances apoptosis in MLL-rearranged pediatric B-acute lymphoblastic leukemia cells
- Author
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Accordi, B, primary, Galla, L, additional, Milani, G, additional, Curtarello, M, additional, Serafin, V, additional, Lissandron, V, additional, Viola, G, additional, te Kronnie, G, additional, De Maria, R, additional, Petricoin, E F, additional, Liotta, L A, additional, Indraccolo, S, additional, and Basso, G, additional
- Published
- 2012
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8. VEGF-targeted therapy stably modulates the glycolytic phenotype of tumor cells
- Author
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Giovanni Esposito, Rosa Maria Moresco, Elisabetta Zulato, Henrike Schroer, Francesco Ciccarese, Matteo Curtarello, Andrea Rasola, Wolfgang Mueller-Klieser, Luca Persano, Stefan Walenta, Mario Plebani, Aichi Msaki, Giulia Guzzo, Stefano Indraccolo, Elisabetta Rossi, Roberta Bertorelle, Anna Pastò, Alberto Amadori, Silvia Valtorta, Sergio Todde, Giorgia Nardo, Curtarello, M, Zulato, E, Nardo, G, Valtorta, S, Guzzo, G, Rossi, E, Esposito, G, Msaki, A, Pastò, A, Rasola, A, Persano, L, Ciccarese, F, Bertorelle, R, Todde, S, Plebani, M, Schroer, H, Walenta, S, Mueller Klieser, W, Amadori, A, Moresco, R, and Indraccolo, S
- Subjects
Vascular Endothelial Growth Factor A ,Cancer Research ,Pathology ,medicine.medical_specialty ,Necrosis ,medicine.medical_treatment ,Angiogenesis Inhibitors ,Mice, SCID ,Biology ,SCID ,Antibodies, Monoclonal, Humanized ,Antibodies ,Cell Line ,Targeted therapy ,Mice ,Random Allocation ,Cell Line, Tumor ,Neoplasms ,Monoclonal ,Animals ,Bevacizumab ,Female ,Glycolysis ,Humans ,MCF-7 Cells ,Mice, Inbred BALB C ,Molecular Targeted Therapy ,Phenotype ,Xenograft Model Antitumor Assays ,medicine ,cancer-cell ,Anti-VEGF therapy ,Humanized ,Inbred BALB C ,MED/36 - DIAGNOSTICA PER IMMAGINI E RADIOTERAPIA ,Tumor ,positron emission tomography, antiangiogenesis, glucose metabolism, hypoxia ,Blockade ,Vascular endothelial growth factor A ,Oncology ,Cell culture ,Cancer research ,MED/06 - ONCOLOGIA MEDICA ,medicine.symptom - Abstract
Anti-VEGF therapy perturbs tumor metabolism, severely impairing oxygen, glucose, and ATP levels. In this study, we investigated the effects of anti-VEGF therapy in multiple experimental tumor models that differ in their glycolytic phenotypes to gain insights into optimal modulation of the metabolic features of this therapy. Prolonged treatments induced vascular regression and necrosis in tumor xenograft models, with highly glycolytic tumors becoming treatment resistant more rapidly than poorly glycolytic tumors. By PET imaging, prolonged treatments yielded an increase in both hypoxic and proliferative regions of tumors. A selection for highly glycolytic cells was noted and this metabolic shift was stable and associated with increased tumor aggressiveness and resistance to VEGF blockade in serially transplanted mice. Our results support the hypothesis that the highly glycolytic phenotype of tumor cells studied in xenograft models, either primary or secondary, is a cell-autonomous trait conferring resistance to VEGF blockade. The finding that metabolic traits of tumors can be selected by antiangiogenic therapy suggests insights into the evolutionary dynamics of tumor metabolism. Cancer Res; 75(1); 120–33. ©2014 AACR.
- Published
- 2015
9. Anti-VEGF therapy selects for clones resistant to glucose starvation in ovarian cancer xenografts.
- Author
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Boso D, Tognon M, Curtarello M, Minuzzo S, Piga I, Brillo V, Lazzarini E, Carlet J, Marra L, Trento C, Rasola A, Masgras I, Caporali L, Del Ben F, Brisotto G, Turetta M, Pastorelli R, Brunelli L, Navaglia F, Esposito G, Grassi A, and Indraccolo S
- Subjects
- Animals, Female, Humans, Mice, Cell Line, Tumor, Clone Cells metabolism, Clone Cells pathology, Mice, Inbred NOD, Mice, SCID, Oxidative Phosphorylation, Xenograft Model Antitumor Assays, Glucose metabolism, Ovarian Neoplasms drug therapy, Ovarian Neoplasms genetics, Ovarian Neoplasms pathology, Vascular Endothelial Growth Factor A antagonists & inhibitors
- Abstract
Background: Genetic and metabolic heterogeneity are well-known features of cancer and tumors can be viewed as an evolving mix of subclonal populations, subjected to selection driven by microenvironmental pressures or drug treatment. In previous studies, anti-VEGF therapy was found to elicit rewiring of tumor metabolism, causing marked alterations in glucose, lactate ad ATP levels in tumors. The aim of this study was to evaluate whether differences in the sensitivity to glucose starvation existed at the clonal level in ovarian cancer cells and to investigate the effects induced by anti-VEGF therapy on this phenotype by multi-omics analysis., Methods: Clonal populations, obtained from both ovarian cancer cell lines (IGROV-1 and SKOV3) and tumor xenografts upon glucose deprivation, were defined as glucose deprivation resistant (GDR) or glucose deprivation sensitive (GDS) clones based on their in vitro behaviour. GDR and GDS clones were characterized using a multi-omics approach, including genetic, transcriptomic and metabolic analysis, and tested for their tumorigenic potential and reaction to anti-angiogenic therapy., Results: Two clonal populations, GDR and GDS, with strikingly different viability following in vitro glucose starvation, were identified in ovarian cancer cell lines. GDR clones survived and overcame glucose starvation-induced stress by enhancing mitochondrial oxidative phosphorylation (OXPHOS) and both pyruvate and lipids uptake, whereas GDS clones were less able to adapt and died. Treatment of ovarian cancer xenografts with the anti-VEGF drug bevacizumab positively selected for GDR clones that disclosed increased tumorigenic properties in NOD/SCID mice. Remarkably, GDR clones were more sensitive than GDS clones to the mitochondrial respiratory chain complex I inhibitor metformin, thus suggesting a potential therapeutic strategy to target the OXPHOS-metabolic dependency of this subpopulation., Conclusion: A glucose-deprivation resistant population of ovarian cancer cells showing druggable OXPHOS-dependent metabolic traits is enriched in experimental tumors treated by anti-VEGF therapy., (© 2023. Italian National Cancer Institute ‘Regina Elena’.)
- Published
- 2023
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10. p53/ TP53 Status Assessment in Gastroesophageal Adenocarcinoma.
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Boldrin E, Piano MA, Bernaudo F, Alfieri R, Biasin MR, Montagner IM, Volpato A, Mattara G, Lamacchia F, Magni G, Rosato A, Scapinello A, Pilati P, and Curtarello M
- Abstract
Chromosomal instability (CIN) is very frequent in gastroesophageal adenocarcinoma (GEA) and it is characterized by TP53 deletions/mutations resulting in p53 nuclear accumulation, as revealed by immunohistochemistry (IHC), which considers the cases with "high" staining levels to be positive. Aiming to improve aberrant TP53 detection, droplet digital PCR (ddPCR) was used to evaluate TP53 deletion in formalin-fixed, paraffin-embedded DNA (FFPE-DNA) and cell-free DNA (cfDNA). To further investigate the mutational TP53 profile, next-generation sequencing (NGS) was performed in a subset of FFPE samples. After combining "low" and "high" IHC staining level groups, the proportion of deletion events was significantly higher compared to the "intermediate" group (72.9% vs. 47.5%, p -value = 0.002). The ddPCR TP53 deletion assay was feasible for cfDNA but only had good agreement (72.7%, Cohen's kappa = 0.48) with the assay performed with FFPE-DNA of the "low-level" group. NGS analysis confirmed that, in the "low-level" group, a high percentage (66.7%) of cases were aberrant, with disruptive mutations that probably led to p53 loss. Data suggested that p53 IHC alone underestimates the CIN phenotype in GEA and that molecular analysis in both solid and liquid biopsies could be integrated with it; in particular, in cases of completely negative staining.
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- 2023
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11. Putative Clinical Potential of ERBB2 Amplification Assessment by ddPCR in FFPE-DNA and cfDNA of Gastroesophageal Adenocarcinoma Patients.
- Author
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Boldrin E, Mazza M, Piano MA, Alfieri R, Montagner IM, Magni G, Scaini MC, Vassallo L, Rosato A, Pilati P, Scapinello A, and Curtarello M
- Abstract
Anti-HER2 monoclonal antibody trastuzumab improves the survival of those patients with advanced gastroesophageal adenocarcinoma (GEA) exhibiting HER2/ ERBB2 overexpression/amplification. The current gold standard methods used to diagnose the HER2 status in GEA are immunohistochemistry (IHC) and silver or fluorescence in situ hybridization (SISH or FISH). However, they do not permit spatial and temporal tumor monitoring, nor do they overcome intra-cancer heterogeneity. Droplet digital PCR (ddPCR) was used to implement the assessment of HER2 status in formalin-fixed paraffin-embedded (FFPE) tumor DNA from a retrospective cohort (86 patients) and in cell-free DNA (cfDNA) samples from a prospective cohort (28 patients). In comparison to IHC/SISH, ddPCR assay revealed ERBB2 amplification in a larger patient fraction, including HER2 2+ and 0-1+ of the retrospective cohort (45.3% vs. 15.1%). In addition, a considerable number of HER2 2+ and 0-1+ prospective patients who were negative in FFPE by both IHC/SISH and ddPCR, showed ERBB2 amplification in the cfDNA collected just before surgery. cfDNA analysis in a few longitudinal cases revealed an increasing ERBB2 trend at progression. In conclusion, ddPCR in liquid biopsy may improve the detection rate of HER2 positive patients, preventing those patients who could benefit from targeted therapy from being incorrectly excluded.
- Published
- 2022
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12. MSI Analysis in Solid and Liquid Biopsies of Gastroesophageal Adenocarcinoma Patients: A Molecular Approach.
- Author
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Boldrin E, Piano MA, Alfieri R, Mazza M, Vassallo L, Scapinello A, Pilati P, and Curtarello M
- Subjects
- Adult, Aged, Aged, 80 and over, Cell-Free Nucleic Acids genetics, Female, Humans, Liquid Biopsy methods, Male, Microsatellite Instability, Middle Aged, Prospective Studies, Retrospective Studies, Adenocarcinoma genetics, Adenocarcinoma pathology, Esophageal Neoplasms genetics, Esophageal Neoplasms pathology, Esophagogastric Junction pathology, Stomach Neoplasms genetics, Stomach Neoplasms pathology
- Abstract
Gastroesophageal adenocarcinoma (GEA) patients with the microsatellite instability (MSI) subtype emerged as optimal candidates for immunotherapy. To date, immunohistochemistry (IHC) is the gold standard for MSI assessment in formalin-fixed paraffin-embedded (FFPE) specimens. However, IHC, although useful for diagnostic typing, cannot be used to analyze cell-free DNA (cfDNA) in liquid biopsy, a tool that could overcome tumor heterogeneity and enable longitudinal monitoring. In order to find an alternative diagnostic method to IHC, we analyzed 86 retrospective GEAs FFPE samples with multiplex PCR. Moreover, to verify the feasibility of MSI detection in liquid biopsy, cfDNA samples of five patients that resulted in having MSI in a prospective cohort of 35 patients were evaluated by multiplex PCR, real-time PCR and droplet digital PCR (ddPCR). Analysis of FFPE showed 100% concordance between multiplex PCR and IHC (Cohen's Kappa agreement = 1). On the contrary, only ddPCR was able to detect MSI in cfDNAs of T3/T4 GEA patients. In conclusion, data highlight the molecular analysis as an optimal alternative to IHC for the diagnostic typing and suggest that the ddPCR assay can be considered as the most reliable and promising molecular approach to detect MSI in the cfDNA of GEA patients.
- Published
- 2021
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13. Genetic Perturbation of Pyruvate Dehydrogenase Kinase 1 Modulates Growth, Angiogenesis and Metabolic Pathways in Ovarian Cancer Xenografts.
- Author
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Venturoli C, Piga I, Curtarello M, Verza M, Esposito G, Venuto S, Navaglia F, Grassi A, and Indraccolo S
- Subjects
- Animals, Cell Line, Tumor, Female, Glycolysis, Heterografts, Humans, Mice, Mice, Inbred NOD, Neovascularization, Pathologic, Ovarian Neoplasms genetics, Pyruvate Dehydrogenase Acetyl-Transferring Kinase metabolism
- Abstract
Pyruvate dehydrogenase kinase 1 (PDK1) blockade triggers are well characterized in vitro metabolic alterations in cancer cells, including reduced glycolysis and increased glucose oxidation. Here, by gene expression profiling and digital pathology-mediated quantification of in situ markers in tumors, we investigated effects of PDK1 silencing on growth, angiogenesis and metabolic features of tumor xenografts formed by highly glycolytic OC316 and OVCAR3 ovarian cancer cells. Notably, at variance with the moderate antiproliferative effects observed in vitro, we found a dramatic negative impact of PDK1 silencing on tumor growth. These findings were associated with reduced angiogenesis and increased necrosis in the OC316 and OVCAR3 tumor models, respectively. Analysis of viable tumor areas uncovered increased proliferation as well as increased apoptosis in PDK1-silenced OVCAR3 tumors. Moreover, RNA profiling disclosed increased glucose catabolic pathways-comprising both oxidative phosphorylation and glycolysis-in PDK1-silenced OVCAR3 tumors, in line with the high mitotic activity detected in the viable rim of these tumors. Altogether, our findings add new evidence in support of a link between tumor metabolism and angiogenesis and remark on the importance of investigating net effects of modulations of metabolic pathways in the context of the tumor microenvironment.
- Published
- 2021
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14. PIK3CA Mutation in the ShortHER Randomized Adjuvant Trial for Patients with Early HER2 + Breast Cancer: Association with Prognosis and Integration with PAM50 Subtype.
- Author
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Guarneri V, Dieci MV, Bisagni G, Brandes AA, Frassoldati A, Cavanna L, Musolino A, Giotta F, Rimanti A, Garrone O, Bertone E, Cagossi K, Nanni O, Piacentini F, Orvieto E, Griguolo G, Curtarello M, Urso L, Paré L, Chic N, D'Amico R, Prat A, and Conte P
- Subjects
- Adult, Aged, Antineoplastic Combined Chemotherapy Protocols administration & dosage, B7-H1 Antigen genetics, Biomarkers, Tumor genetics, Breast Neoplasms genetics, Breast Neoplasms pathology, CD8 Antigens genetics, Cell Cycle Proteins genetics, Disease-Free Survival, Female, Humans, Kaplan-Meier Estimate, Lymphocytes, Tumor-Infiltrating drug effects, Middle Aged, Mutation genetics, Prognosis, Programmed Cell Death 1 Receptor genetics, Receptors, Estrogen genetics, Trans-Activators genetics, Trastuzumab adverse effects, Breast Neoplasms drug therapy, Class I Phosphatidylinositol 3-Kinases genetics, Receptor, ErbB-2 genetics, Trastuzumab administration & dosage
- Abstract
Purpose: We explored the prognostic effect of PIK3CA mutation in HER2
+ patients enrolled in the ShortHER trial., Patients and Methods: The ShortHER trial randomized 1,253 patients with HER2+ breast cancer to 9 weeks or 1 year of adjuvant trastuzumab combined with chemotherapy. PIK3CA hotspot mutations in exon 9 and 20 were analyzed by pyrosequencing. Expression of 60 genes, including PAM50 genes was measured using the nCounter platform., Results: A mutation of the PIK3CA gene was detected in 21.7% of the 803 genotyped tumors. At a median follow-up of 7.7 years, 5-year disease-free survival (DFS) rates were 90.6% for PIK3CA mutated and 86.2% for PIK3CA wild-type tumors [HR, 0.84; 95% confidence interval (CI), 0.56-1.27; P = 0.417]. PIK3CA mutation showed a favorable prognostic impact in the PAM50 HER2-enriched subtype ( n = 232): 5-year DFS 91.8% versus 76.1% (log-rank P = 0.049; HR, 0.46; 95% CI, 0.21-1.02). HER2-enriched/ PIK3CA mutated versus wild-type tumors showed numerically higher tumor-infiltrating lymphocytes (TIL) and significant upregulation of immune-related genes (including CD8A, CD274, PDCD1, and MYBL2 , a proliferation gene involved in immune processes). High TILs as well as the upregulation of PDCD1 and MYBL2 were associated with a significant DFS improvement within the HER2-enriched subtype (HR, 0.82; 95% CI, 0.68-0.99; P = 0.039 for 10% TILs increment; HR, 0.81; 95% CI, 0.65-0.99; P = 0.049 for PDCD1 expression; HR, 0.72; 95% CI, 0.53-0.99; P = 0.042 for MYBL2 expression)., Conclusions: PIK3CA mutation showed no prognostic impact in the ShortHER trial. Within the HER2-enriched molecular subtype, patients with PIK3CA mutated tumors showed better DFS versus PIK3CA wild-type, which may be partly explained by upregulation of immune-related genes., (©2020 American Association for Cancer Research.)- Published
- 2020
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15. Detection of LINE-1 hypomethylation in cfDNA of Esophageal Adenocarcinoma Patients.
- Author
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Boldrin E, Curtarello M, Dallan M, Alfieri R, Realdon S, Fassan M, and Saggioro D
- Subjects
- Adenocarcinoma pathology, Adult, Aged, Barrett Esophagus pathology, Disease Progression, Esophageal Neoplasms pathology, Feasibility Studies, Female, Humans, Male, Middle Aged, Monitoring, Physiologic methods, Adenocarcinoma genetics, Barrett Esophagus genetics, Cell-Free Nucleic Acids genetics, DNA Methylation, Esophageal Neoplasms genetics, Long Interspersed Nucleotide Elements genetics
- Abstract
DNA methylation plays an important role in cancer development. Cancer cells exhibit two types of DNA methylation alteration: site-specific hypermethylation at promoter of oncosuppressor genes and global DNA hypomethylation. This study evaluated the methylation patterns of long interspersed nuclear element (LINE-1) sequences which, due to their relative abundance in the genome, are considered a good surrogate indicator of global DNA methylation. LINE-1 methylation status was investigated in the cell-free DNA (cfDNA) of 21 patients, 19 with esophageal adenocarcinoma (EADC) and 2 with Barrett's esophagus (BE). The two BE patients and one EADC patient were also analyzed longitudinally. Methylation status was analyzed using restriction enzymes and DNA amplification. This methodology was chosen to avoid bisulfite conversion, which we considered inadequate for cfDNA analysis. Indeed, cfDNA is characterized by poor quality and low concentration, and bisulfite conversion might worsen these conditions. Results showed that hypomethylated LINE-1 sequences are present in EADC cfDNA. Furthermore, longitudinal studies in BE suggested a correlation between methylation status of LINE-1 sequences in cfDNA and progression to EADC. In conclusion, our study indicated the feasibility of our methodological approach to detect hypomethylation events in cfDNA from EADC patients, and suggests LINE-1 methylation analysis as a new possible molecular assay to integrate into patient monitoring.
- Published
- 2020
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16. Rewiring of Lipid Metabolism and Storage in Ovarian Cancer Cells after Anti-VEGF Therapy.
- Author
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Curtarello M, Tognon M, Venturoli C, Silic-Benussi M, Grassi A, Verza M, Minuzzo S, Pinazza M, Brillo V, Tosi G, Ferrazza R, Guella G, Iorio E, Godfroid A, Sounni NE, Amadori A, and Indraccolo S
- Subjects
- Apoptosis drug effects, Benzoates therapeutic use, Benzylamines therapeutic use, Bevacizumab therapeutic use, Blotting, Western, Cell Line, Tumor, Cell Proliferation drug effects, Cell Survival drug effects, Female, Humans, Immunohistochemistry, Magnetic Resonance Spectroscopy, Vascular Endothelial Growth Factor A antagonists & inhibitors, Xenograft Model Antitumor Assays, Lipid Metabolism drug effects, Ovarian Neoplasms drug therapy, Ovarian Neoplasms metabolism, Vascular Endothelial Growth Factor A metabolism
- Abstract
Anti-angiogenic therapy triggers metabolic alterations in experimental and human tumors, the best characterized being exacerbated glycolysis and lactate production. By using both Liquid Chromatography-Mass Spectrometry (LC-MS) and Nuclear Magnetic Resonance (NMR) analysis, we found that treatment of ovarian cancer xenografts with the anti-Vascular Endothelial Growth Factor (VEGF) neutralizing antibody bevacizumab caused marked alterations of the tumor lipidomic profile, including increased levels of triacylglycerols and reduced saturation of lipid chains. Moreover, transcriptome analysis uncovered up-regulation of pathways involved in lipid metabolism. These alterations were accompanied by increased accumulation of lipid droplets in tumors. This phenomenon was reproduced under hypoxic conditions in vitro, where it mainly depended from uptake of exogenous lipids and was counteracted by treatment with the Liver X Receptor (LXR)-agonist GW3965, which inhibited cancer cell viability selectively under reduced serum conditions. This multi-level analysis indicates alterations of lipid metabolism following anti-VEGF therapy in ovarian cancer xenografts and suggests that LXR-agonists might empower anti-tumor effects of bevacizumab.
- Published
- 2019
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17. Silencing of miR-182 is associated with modulation of tumorigenesis through apoptosis induction in an experimental model of colorectal cancer.
- Author
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Perilli L, Tessarollo S, Albertoni L, Curtarello M, Pastò A, Brunetti E, Fassan M, Rugge M, Indraccolo S, Amadori A, Bortoluzzi S, and Zanovello P
- Subjects
- Animals, Caco-2 Cells, Cell Cycle genetics, Colorectal Neoplasms pathology, Disease Models, Animal, Gene Expression Regulation, Neoplastic, HT29 Cells, Heterografts, Humans, Mice, Mice, Inbred NOD, Mice, SCID, Transfection, Tumor Burden genetics, Up-Regulation genetics, Apoptosis genetics, Carcinogenesis genetics, Colorectal Neoplasms genetics, Gene Silencing, MicroRNAs genetics
- Abstract
Background: miR-182-5p (miR-182) is an oncogenic microRNA (miRNA) found in different tumor types and one of the most up-regulated miRNA in colorectal cancer (CRC). Although this microRNA is expressed in the early steps of tumor development, its role in driving tumorigenesis is unclear., Methods: The effects of miR-182 silencing on transcriptomic profile were investigated using two CRC cell lines characterized by different in vivo biological behavior, the MICOL-14
h-tert cell line (dormant upon transfer into immunodeficient hosts) and its tumorigenic variant, MICOL-14tum . Apoptosis was studied by annexin/PI staining and cleaved Caspase-3/PARP analysis. The effect of miR-182 silencing on the tumorigenic potential was addressed in a xenogeneic model of MICOL-14tum transplant., Results: Endogenous miR-182 expression was higher in MICOL-14tum than in MICOL-14h-tert cells. Interestingly, miR-182 silencing had a strong impact on gene expression profile, and the positive regulation of apoptotic process was one of the most affected pathways. Accordingly, annexin/PI staining and caspase-3/PARP activation demonstrated that miR-182 treatment significantly increased apoptosis, with a prominent effect in MICOL-14tum cells. Moreover, a significant modulation of the cell cycle profile was exerted by anti-miR-182 treatment only in MICOL-14tum cells, where a significant increase in the fraction of cells in G0/G1 phases was observed. Accordingly, a significant growth reduction and a less aggressive histological aspect were observed in tumor masses generated by in vivo transfer of anti-miR-182-treated MICOL-14tum cells into immunodeficient hosts., Conclusions: Altogether, these data indicate that increased miR-182 expression may promote cell proliferation, suppress the apoptotic pathway and ultimately confer aggressive traits on CRC cells.- Published
- 2019
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18. Absence of Neurofibromin Induces an Oncogenic Metabolic Switch via Mitochondrial ERK-Mediated Phosphorylation of the Chaperone TRAP1.
- Author
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Masgras I, Ciscato F, Brunati AM, Tibaldi E, Indraccolo S, Curtarello M, Chiara F, Cannino G, Papaleo E, Lambrughi M, Guzzo G, Gambalunga A, Pizzi M, Guzzardo V, Rugge M, Vuljan SE, Calabrese F, Bernardi P, and Rasola A
- Subjects
- Animals, CRISPR-Cas Systems genetics, Cell Line, HSP90 Heat-Shock Proteins antagonists & inhibitors, HSP90 Heat-Shock Proteins genetics, Humans, Hypoxia-Inducible Factor 1, alpha Subunit metabolism, MAP Kinase Kinase 1 genetics, MAP Kinase Kinase 1 metabolism, Mice, Molecular Dynamics Simulation, Neurofibromin 1 deficiency, Phosphorylation, Protein Structure, Tertiary, RNA Interference, RNA, Small Interfering metabolism, Signal Transduction drug effects, Succinate Dehydrogenase antagonists & inhibitors, Succinate Dehydrogenase metabolism, Succinates chemistry, Succinates pharmacology, ras Proteins metabolism, HSP90 Heat-Shock Proteins metabolism, Mitochondria metabolism, Mitogen-Activated Protein Kinase 1 metabolism, Mitogen-Activated Protein Kinase 3 metabolism, Neurofibromin 1 genetics
- Abstract
Mutations in neurofibromin, a Ras GTPase-activating protein, lead to the tumor predisposition syndrome neurofibromatosis type 1. Here, we report that cells lacking neurofibromin exhibit enhanced glycolysis and decreased respiration in a Ras/ERK-dependent way. In the mitochondrial matrix of neurofibromin-deficient cells, a fraction of active ERK1/2 associates with succinate dehydrogenase (SDH) and TRAP1, a chaperone that promotes the accumulation of the oncometabolite succinate by inhibiting SDH. ERK1/2 enhances both formation of this multimeric complex and SDH inhibition. ERK1/2 kinase activity is favored by the interaction with TRAP1, and TRAP1 is, in turn, phosphorylated in an ERK1/2-dependent way. TRAP1 silencing or mutagenesis at the serine residues targeted by ERK1/2 abrogates tumorigenicity, a phenotype that is reverted by addition of a cell-permeable succinate analog. Our findings reveal that Ras/ERK signaling controls the metabolic changes orchestrated by TRAP1 that have a key role in tumor growth and are a promising target for anti-neoplastic strategies., (Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
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19. A hypoxic signature marks tumors formed by disseminated tumor cells in the BALB-neuT mammary cancer model.
- Author
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Msaki A, Pastò A, Curtarello M, Arigoni M, Barutello G, Calogero RA, Macagno M, Cavallo F, Amadori A, and Indraccolo S
- Subjects
- Animals, Carcinogenesis, Cell Hypoxia physiology, Disease Progression, Female, Mammary Neoplasms, Experimental genetics, Mammary Neoplasms, Experimental pathology, Mice, Mice, Inbred BALB C, Mice, Transgenic, Mammary Neoplasms, Experimental metabolism
- Abstract
Metastasis is the final stage of cancer progression. Some evidence indicates that tumor cell dissemination occurs early in the natural history of cancer progression. Disseminated tumor cells (DTC) have been described in the bone marrow (BM) of cancer patients as well as in experimental models, where they correlate with later development of metastasis. However, little is known about the tumorigenic features of DTC obtained at different time points along tumor progression. Here, we found that early DTC isolated from BM of 15-17 week-old Her2/neu transgenic (BALB-neuT) mice were not tumorigenic in immunodeficient mice. In contrast, DTC-derived tumors were easily detectable when late DTC obtained from 19-22 week-old BALB-neuT mice were injected. Angiogenesis, which contributes to regulate tumor dormancy, appeared dispensable to reactivate late DTC, although it accelerated growth of secondary DTC tumors. Compared with parental mammary tumors, gene expression profiling disclosed a distinctive transcriptional signature of late DTC tumors which was enriched for hypoxia-related transcripts and was maintained in ex-vivo cell culture. Altogether, these findings highlight a different tumorigenic potential of early and late DTC in the BALB-neuT model and describe a HIF-1α-related transcriptional signature in DTC tumors, which may render DTC angiogenesis-competent, when placed in a favourable environment., Competing Interests: The authors declare no conflicts of interest.
- Published
- 2016
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20. VEGF-targeted therapy stably modulates the glycolytic phenotype of tumor cells.
- Author
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Curtarello M, Zulato E, Nardo G, Valtorta S, Guzzo G, Rossi E, Esposito G, Msaki A, Pastò A, Rasola A, Persano L, Ciccarese F, Bertorelle R, Todde S, Plebani M, Schroer H, Walenta S, Mueller-Klieser W, Amadori A, Moresco RM, and Indraccolo S
- Subjects
- Animals, Antibodies, Monoclonal, Humanized pharmacology, Bevacizumab, Cell Line, Tumor, Female, Glycolysis drug effects, Humans, MCF-7 Cells, Mice, Mice, Inbred BALB C, Mice, SCID, Molecular Targeted Therapy, Neoplasms blood supply, Phenotype, Random Allocation, Vascular Endothelial Growth Factor A metabolism, Xenograft Model Antitumor Assays, Angiogenesis Inhibitors pharmacology, Neoplasms drug therapy, Neoplasms metabolism, Vascular Endothelial Growth Factor A antagonists & inhibitors
- Abstract
Anti-VEGF therapy perturbs tumor metabolism, severely impairing oxygen, glucose, and ATP levels. In this study, we investigated the effects of anti-VEGF therapy in multiple experimental tumor models that differ in their glycolytic phenotypes to gain insights into optimal modulation of the metabolic features of this therapy. Prolonged treatments induced vascular regression and necrosis in tumor xenograft models, with highly glycolytic tumors becoming treatment resistant more rapidly than poorly glycolytic tumors. By PET imaging, prolonged treatments yielded an increase in both hypoxic and proliferative regions of tumors. A selection for highly glycolytic cells was noted and this metabolic shift was stable and associated with increased tumor aggressiveness and resistance to VEGF blockade in serially transplanted mice. Our results support the hypothesis that the highly glycolytic phenotype of tumor cells studied in xenograft models, either primary or secondary, is a cell-autonomous trait conferring resistance to VEGF blockade. The finding that metabolic traits of tumors can be selected by antiangiogenic therapy suggests insights into the evolutionary dynamics of tumor metabolism., (©2014 American Association for Cancer Research.)
- Published
- 2015
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21. PKH26 staining defines distinct subsets of normal human colon epithelial cells at different maturation stages.
- Author
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Pastò A, Marchesi M, Diamantini A, Frasson C, Curtarello M, Lago C, Pilotto G, Parenti AR, Esposito G, Agostini M, Nitti D, and Amadori A
- Subjects
- Biomarkers metabolism, Cell Differentiation, Cell Proliferation, Gene Expression Regulation, Humans, Intestinal Mucosa metabolism, Spheroids, Cellular cytology, Spheroids, Cellular metabolism, Staining and Labeling, Cell Separation methods, Colon cytology, Intestinal Mucosa cytology, Organic Chemicals metabolism
- Abstract
Background and Aim: Colon crypts are characterized by a hierarchy of cells distributed along the crypt axis. Aim of this paper was to develop an in vitro system for separation of epithelial cell subsets in different maturation stages from normal human colon., Methodology and Major Findings: Dissociated colonic epithelial cells were stained with PKH26, which allows identification of distinct populations based on their proliferation rate, and cultured in vitro in the absence of serum. The cytofluorimetric expression of CK20, Msi-1 and Lgr5 was studied. The mRNA levels of several stemness-associated genes were also compared in cultured cell populations and in three colon crypt populations isolated by microdissection. A PKH(pos) population survived in culture and formed spheroids; this population included subsets with slow (PKH(high)) and rapid (PKH(low)) replicative rates. Molecular analysis revealed higher mRNA levels of both Msi-1 and Lgr-5 in PKH(high) cells; by cytofluorimetric analysis, Msi-1(+)/Lgr5(+) cells were only found within PKH(high) cells, whereas Msi-1(+)/Lgr5(-) cells were also observed in the PKH(low) population. As judged by qRT-PCR analysis, the expression of several stemness-associated markers (Bmi-1, EphB2, EpCAM, ALDH1) was highly enriched in Msi-1(+)/Lgr5(+) cells. While CK20 expression was mainly found in PKH(low) and PKH(neg) cells, a small PKH(high) subset co-expressed both CK20 and Msi-1, but not Lgr5; cells with these properties also expressed Mucin, and could be identified in vivo in colon crypts. These results mirrored those found in cells isolated from different crypt portions by microdissection, and based on proliferation rates and marker expression they allowed to define several subsets at different maturation stages: PKH(high)/Lgr5(+)/Msi-1(+)/CK20(-), PKH(high)/Lgr5(-)/Msi-1(+)/CK20(+), PKH(low)/Lgr5(-)/Msi-1(+)/Ck20(-), and PKH(low)/Lgr5(-)/Msi-1(-)/CK20(+) cells., Conclusions: Our data show the possibility of deriving in vitro, without any selection strategy, several distinct cell subsets of human colon epithelial cells, which recapitulate the phenotypic and molecular profile of cells in a discrete crypt location.
- Published
- 2012
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22. Glycolytic phenotype and AMP kinase modify the pathologic response of tumor xenografts to VEGF neutralization.
- Author
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Nardo G, Favaro E, Curtarello M, Moserle L, Zulato E, Persano L, Rossi E, Esposito G, Crescenzi M, Casanovas O, Sattler U, Mueller-Klieser W, Biesalski B, Thews O, Canese R, Iorio E, Zanovello P, Amadori A, and Indraccolo S
- Subjects
- Animals, Cell Line, Tumor, Fluorodeoxyglucose F18 pharmacokinetics, Humans, Magnetic Resonance Imaging, Magnetic Resonance Spectroscopy, Mice, Neoplasms, Experimental metabolism, Neoplasms, Experimental pathology, Phenotype, Adenylate Kinase physiology, Glycolysis, Neoplasms, Experimental drug therapy, Vascular Endothelial Growth Factor A antagonists & inhibitors
- Abstract
VEGF antagonists are now widely used cancer therapeutics, but predictive biomarkers of response or toxicity remain unavailable. In this study, we analyzed the effects of anti-VEGF therapy on tumor metabolism and therapeutic response by using an integrated set of imaging techniques, including bioluminescence metabolic imaging, 18-fluorodeoxyglucose positron emission tomography, and MRI imaging and spectroscopy. Our results revealed that anti-VEGF therapy caused a dramatic depletion of glucose and an exhaustion of ATP levels in tumors, although glucose uptake was maintained. These metabolic changes selectively accompanied the presence of large necrotic areas and partial tumor regression in highly glycolytic tumors. In addition, we found that the central metabolic protein kinase AMP-activated protein kinase (AMPK)-a cellular sensor of ATP levels that supports cell viability in response to energy stress-was activated by anti-VEGF therapy in experimental tumors. AMPK-α2 attenuation increased glucose consumption, tumor cell sensitivity to glucose starvation, and tumor necrosis following anti-VEGF therapy. Taken together, our findings reveal functional links between the Warburg effect and the AMPK pathway with therapeutic responses to VEGF neutralization in tumor xenograft models.
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- 2011
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23. Lentiviral-mediated RNAi in vivo silencing of Col6a1, a gene with complex tissue specific expression pattern.
- Author
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Frka K, Facchinello N, Del Vecchio C, Carpi A, Curtarello M, Venerando R, Angelin A, Parolin C, Bernardi P, Bonaldo P, Volpin D, Braghetta P, and Bressan GM
- Subjects
- Animals, Apoptosis, Blotting, Northern, Cell Line, Collagen Type VI genetics, Female, Gene Expression Profiling, Genetic Vectors genetics, Immunoblotting, Male, Membrane Potential, Mitochondrial, Mice, Mice, Transgenic, NIH 3T3 Cells, Reverse Transcriptase Polymerase Chain Reaction, Collagen Type VI metabolism, Lentivirus genetics, RNA Interference
- Abstract
RNA interference (RNAi) through the use of lentiviral vectors is a valuable technique to induce loss of function mutations in mammals. Although very promising, the method has found only limited application and its general applicability remains to be established. Here we analyze how different factors influence RNAi mediated silencing of Col6a1, a gene of the extracellular matrix with a complex pattern of tissue specific expression. Our results, obtained with vectors pLVTHM and pLVPT-rtTRKRAB, point out three parameters as major determinants of the efficiency of interference: the choice of interfering sequence, the number of proviral copies integrated into the mouse genome and the site of insertion of the provirus. Although low copy number may produce efficient interference with low frequency, the general trend is that the number of integrated proviral copies determines the level of silencing and the severity of phenotypic traits. The site of insertion not only determines the overall intensity of expression of the small interfering RNA (siRNA), but also introduces slight variability of silencing in different organs. A lentiviral vector (pLVPT-rtTRKRAB) with doxycycline-inducible production of siRNA was also tested. Control of expression by the drug was stringent in many tissues; however, in some tissues turning off of siRNA synthesis was not complete. The data support the application of lentiviral vectors used here in transgenesis.
- Published
- 2009
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