145 results on '"Tommaso Mello"'
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2. Supplementary Figure Legends from DNA Damage Response Protein CHK2 Regulates Metabolism in Liver Cancer
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Vinicio Carloni, Andrea Galli, Krista Rombouts, Antonio Mazzocca, Francesco Paolo Fanizzi, Paolo Forte, Laura Gragnani, Stefania Madiai, Caecilia Sukowati, Tommaso Mello, Laura Del Coco, and Matteo Lulli
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Description of supplementary figures
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- 2023
3. Table S1 from DNA Damage Response Protein CHK2 Regulates Metabolism in Liver Cancer
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Vinicio Carloni, Andrea Galli, Krista Rombouts, Antonio Mazzocca, Francesco Paolo Fanizzi, Paolo Forte, Laura Gragnani, Stefania Madiai, Caecilia Sukowati, Tommaso Mello, Laura Del Coco, and Matteo Lulli
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Table S1. Characteristics of the healthy subjects (HS) and patients with liver cirrhosis (LC) and hepatocellular carcinoma (HCC)
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- 2023
4. Table S3 from DNA Damage Response Protein CHK2 Regulates Metabolism in Liver Cancer
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Vinicio Carloni, Andrea Galli, Krista Rombouts, Antonio Mazzocca, Francesco Paolo Fanizzi, Paolo Forte, Laura Gragnani, Stefania Madiai, Caecilia Sukowati, Tommaso Mello, Laura Del Coco, and Matteo Lulli
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Table S3. % 13C enrichment in cell lysates for13C-alanine (mitochondrial pyruvate labeling) and 13C-lactate (cytosol pyruvate labeling) obtained by 1H NMR analysis.
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- 2023
5. A mutation in the ZNF687 gene that is responsible for the severe form of Paget’s disease of bone causes severely altered bone remodeling and promotes hepatocellular carcinoma onset in a knock-in mouse model
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Sharon Russo, Federica Scotto di Carlo, Antonio Maurizi, Giorgio Fortunato, Anna Teti, Danilo Licastro, Carmine Settembre, Tommaso Mello, Fernando Gianfrancesco, Russo, Sharon, Scotto di Carlo, Federica, Maurizi, Antonio, Fortunato, Giorgio, Teti, Anna, Licastro, Danilo, Settembre, Carmine, Mello, Tommaso, and Gianfrancesco, Fernando
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Bone, Bone cancer ,Histology ,Physiology ,Endocrinology, Diabetes and Metabolism - Abstract
Paget’s disease (PDB) is a late-onset bone remodeling disorder with a broad spectrum of symptoms and complications. One of the most aggressive forms is caused by the P937R mutation in the ZNF687 gene. Although the genetic involvement of ZNF687 in PDB has been extensively studied, the molecular mechanisms underlying this association remain unclear. Here, we describe the first Zfp687 knock-in mouse model and demonstrate that the mutation recapitulates the PDB phenotype, resulting in severely altered bone remodeling. Through microcomputed tomography analysis, we observed that 8-month-old mutant mice showed a mainly osteolytic phase, with a significant decrease in the trabecular bone volume affecting the femurs and the vertebrae. Conversely, osteoblast activity was deregulated, producing disorganized bone. Notably, this phenotype became pervasive in 16-month-old mice, where osteoblast function overtook bone resorption, as highlighted by the presence of woven bone in histological analyses, consistent with the PDB phenotype. Furthermore, we detected osteophytes and intervertebral disc degeneration, outlining for the first time the link between osteoarthritis and PDB in a PDB mouse model. RNA sequencing of wild-type and Zfp687 knockout RAW264.7 cells identified a set of genes involved in osteoclastogenesis potentially regulated by Zfp687, e.g., Tspan7, Cpe, Vegfc, and Ggt1, confirming its role in this process. Strikingly, in this mouse model, the mutation was also associated with a high penetrance of hepatocellular carcinomas. Thus, this study established an essential role of Zfp687 in the regulation of bone remodeling, offering the potential to therapeutically treat PDB, and underlines the oncogenic potential of ZNF687.
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- 2023
6. Glyco-Coated CdSe/ZnS Quantum Dots as Nanoprobes for Carbonic Anhydrase IX Imaging in Cancer Cells
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Andrea Angeli, Silvia Mussi, Marcello Marelli, Stefano Cicchi, Carla Ghelardini, Claudiu T. Supuran, Arianna Giacomini, Giacomo Biagiotti, Roberto Ronca, Tommaso Mello, Barbara Richichi, Gianluca Salerno, Luca Landini, Lorenzo Di Cesare Mannelli, Gianluca Toniolo, Enzo Menna, and Cosetta Ravelli
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Chemistry ,Quantum dot ,carbonic anhydrase ,glyco-quantum dots ,sulfonamide ,Cancer cell ,Biophysics ,bioimaging ,bladder cancer ,General Materials Science ,Carbonic Anhydrase IX ,Article - Abstract
The bioimaging of cancer cells by the specific targeting of overexpressed biomarkers is an approach that holds great promise in the identification of selective diagnostic tools. Tumor-associated human carbonic anhydrase (hCA) isoforms IX and XII have been considered so far as well-defined biomarkers, with their expression correlating with cancer progression and aggressiveness. Therefore, the availability of highly performant fluorescent tools tailored for their targeting and able to efficiently visualize such key targets is in high demand. We report here on the design and synthesis of a kind of quantum dot (QD)-based fluorescent glyconanoprobe coated with a binary mixture of ligands, which, according to the structure of the terminal domains, impart specific property sets to the fluorescent probe. Specifically, monosaccharide residues ensured the dispersibility in the biological medium, CA inhibitor residues provided specific targeting of membrane-anchored hCA IX overexpressed on bladder cancer cells, and the quantum dots imparted the optical/fluorescence properties.
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- 2021
7. Metformin Treatment Induces Different Response in Pheochromocytoma/Paraganglioma Tumour Cells and in Primary Fibroblasts
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Serena Martinelli, Francesca Amore, Tommaso Mello, Massimo Mannelli, Mario Maggi, and Elena Rapizzi
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Cancer Research ,Oncology ,pheochromocytoma/paraganglioma ,tumour microenvironment ,metformin ,tumour metabolism - Abstract
Pheochromocytoma/paragangliomas (PPGLs) are neuroendocrine tumours, often non-metastatic, but without available effective treatment for their metastatic form. Recent studies have shown that metformin exhibits antiproliferative activity in many human cancers, including PPGLs. Nevertheless, no data are available on the role of metformin on PPGL cells (two-dimension, 2D) and spheroids (three-dimension, 3D) migration/invasion. In this study, we observed that metformin exerts an antiproliferative effect on 2D and 3D cultures of pheochromocytoma mouse tumour tissue (MTT), either silenced or not for the SDHB subunit. However, metformin did not affect MTT migration. On the other hand, metformin did not have a short-term effect on the proliferation of mouse primary fibroblasts, but significantly decreased their ability to migrate. Although the metabolic changes induced by metformin were similar between MTT and fibroblasts (i.e., an overall decrease of ATP production and an increase in intracellular lactate concentration) the activated signalling pathways were different. Indeed, after metformin administration, MTT showed a reduced phosphorylation of Akt and Erk1/2, while fibroblasts exhibited a downregulation of N-Cadherin and an upregulation of E-Cadherin. Herein, we demonstrated that metformin has different effects on cell growth and spread depending on the cell type nature, underlining the importance of the tumour microenvironment in dictating the drug response.
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- 2022
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8. Treatment potential of LPCN 1144 on liver health and metabolic regulation in a non-genomic, high fat diet induced NASH rabbit model
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Tommaso Mello, Andrea Galli, Mario Maggi, Paolo Comeglio, Ilaria Cellai, Benjamin J Bruno, Sandra Filippi, Elisa Maseroli, Giulia Guarnieri, K. Papangkorn, Erica Sarchielli, K. Vangara, Sarah Cipriani, Annamaria Morelli, N. Chidambaram, Giulia Rastrelli, K. Kim, Linda Vignozzi, and M. V. Patel
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Male ,0301 basic medicine ,medicine.medical_specialty ,Steatosis ,Endocrinology, Diabetes and Metabolism ,030209 endocrinology & metabolism ,Inflammation ,Diet, High-Fat ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Non-alcoholic Fatty Liver Disease ,Fibrosis ,Trichrome ,Internal medicine ,Animals ,Medicine ,Prodrugs ,Testosterone ,Metabolic Syndrome ,business.industry ,digestive, oral, and skin physiology ,NASH ,food and beverages ,nutritional and metabolic diseases ,medicine.disease ,Disease Models, Animal ,030104 developmental biology ,Liver ,Androgens ,Biomarker (medicine) ,lipids (amino acids, peptides, and proteins) ,Original Article ,Rabbits ,Steatohepatitis ,medicine.symptom ,business ,Hepatic fibrosis ,hormones, hormone substitutes, and hormone antagonists - Abstract
Purpose Low free testosterone (T) level in men is independently associated with presence and severity of Non-Alcoholic Steatohepatitis (NASH). The histological and molecular effects of oral testosterone prodrug LPCN 1144 treatment on hepatic fibrosis and NASH features are unknown. A metabolic syndrome-induced NASH model in rabbits consuming high fat diet (HFD) has been previously used to assess treatment effects of injectable T on hepatic fibrosis and NASH features. Here we present results on LPCN 1144 in this HFD-induced, NASH preclinical model. Methods Male rabbits were randomly assigned to five groups: regular diet (RD), HFD, HFD + 1144 vehicle (HFD + Veh), HFD + 1144 (1144), and HFD + 1144 + α-tocopherol (1144 + ALPHA). Rabbits were sacrificed after 12 weeks for liver histological, biochemical and genetic analyses. Histological scores were obtained through Giemsa (inflammation), Masson’s trichrome (steatosis and ballooning), and Picrosirius Red (fibrosis) staining. Results Compared to RD, HFD and HFD + Veh significantly worsened NASH features and hepatic fibrosis. Considering HFD and HFD + Veh arms, histological and biomarker features were not significantly different. Both 1144 and 1144 + ALPHA arms improved mean histological scores of NASH as compared to HFD arm. Importantly, percentage of fibrosis was improved in both 1144 (p p = 0.05) treatment arms vs. HFD. Both treatment arms also reduced HFD-induced inflammation and fibrosis mRNA markers. Furthermore, 1144 treatments significantly improved HFD-induced metabolic dysfunctions. Conclusions Histological and biomarker analyses demonstrate that LPCN 1144 improved HFD-induced hepatic fibrosis and NASH biochemical, biomolecular and histochemical features. These preclinical findings support a therapeutic potential of LPCN 1144 in the treatment of NASH and of hepatic fibrosis.
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- 2021
9. Isoforms of the orphan nuclear receptor COUP‑TFII differentially modulate pancreatic cancer progression
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Simone Polvani, Sara Pepe, Sara Tempesti, Mirko Tarocchi, Giada Marroncini, Lapo Bencini, Elisabetta Ceni, Tommaso Mello, Lucia Picariello, Irene Simeone, Cecilia Grappone, Gabriele Dragoni, Lorenzo Antonuzzo, Elisa Giommoni, Stefano Milani, and Andrea Galli
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COUP Transcription Factor II ,Pancreatic Neoplasms ,Mice ,Cancer Research ,Epithelial-Mesenchymal Transition ,Oncology ,Animals ,Humans ,Protein Isoforms ,Orphan Nuclear Receptors ,Carcinoma, Pancreatic Ductal - Abstract
The expression of the nuclear receptor transcription factor (TF) COUP‑TFII is broadly associated with cell differentiation and cancer development, including of pancreatic ductal adenocarcinoma (PDAC), a devastating disease with one of the poorest prognoses among cancers worldwide. Recent studies have started to investigate the pathological and physiological roles of a novel COUP‑TFII isoform (COUP‑TFII_V2) that lacks the DNA‑binding domain. As the role of the canonical COUP‑TFII in PDAC was previously demonstrated, the present study evaluated whether COUP‑TFII_V2 may have a functional role in PDAC. It was demonstrated that COUP‑TFII_V2 naturally occurs in PDAC cells and in primary samples, where its expression is consistent with shorter overall survival and peripheral invasion. Of note, COUP‑TFII_V2, exhibiting nuclear and cytosolic expression, is linked to epithelial to mesenchymal transition (EMT) and cancer progression, as confirmed by nude mouse experiments. The present results demonstrated that COUP‑TFII_V2 distinctively regulates the EMT of PDAC and, similarly to its sibling, it is associated with tumor aggressiveness. The two isoforms have both overlapping and exclusive functions that cooperate with cancer growth and dissemination. By studying how PDAC cells switch from one isoform to the other, novel insight into cancer biology was gained, indicating that this receptor may serve as a novel possible target for PDAC management.
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- 2022
10. RuvBL1 haploinsufficiency improves mTOR-driven NASH-HCC development in mice
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Alice Guida, Irene Simeone, Simone Polvani, Elisabetta Ceni, Lucia Picariello, Gabriele Dragoni, Andrea Galli, and Tommaso Mello
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Hepatology - Published
- 2022
11. SDHB and SDHD silenced pheochromocytoma spheroids respond differently to tumour microenvironment and their aggressiveness is inhibited by impairing stroma metabolism
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Serena Martinelli, Maria Riverso, Tommaso Mello, Francesca Amore, Matteo Parri, Irene Simeone, Massimo Mannelli, Mario Maggi, and Elena Rapizzi
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Paraganglioma ,Succinate Dehydrogenase ,Endocrinology ,Adrenal Gland Neoplasms ,Tumor Microenvironment ,Humans ,Pheochromocytoma ,Molecular Biology ,Biochemistry ,Germ-Line Mutation - Abstract
Germline mutations in more than 20 genes, including those encoding for the succinate dehydrogenase (SDH), predispose to rare tumours, such as pheochromocytoma/paraganglioma (PPGL). Despite encoding for the same enzymatic complex, SDHC and SDHD mutated PHEO/PGLs are generally benign, while up to 80% of SDHB mutated ones are malignant. In this study, we evaluated the different effects of tumour microenvironment on tumour cell migration/invasion, by co-culturing SDHB or SDHD silenced tumour spheroids with primary cancer-associated fibroblasts (CAFs). We observed that SDHD silenced spheroids had an intermediate migration pattern, compared to the highest migration capability of SDHB and the lowest one of the wild type (Wt) spheroids. Interestingly, we noticed that co-culturing Wt, SDHB and SDHD silenced spheroids with CAFs in low glucose (1 g/l) medium, caused a decreased migration of all the spheroids, but only for SDHB silenced ones this reduction was significant. Moreover, the collective migration, observed in high glucose (4.5 g/l) and characteristic of the SDHB silenced cells, was completely lost in low glucose. Importantly, migration could not be recovered even adding glucose (3.5 g/l) to low glucose conditioned medium. When we investigated cell metabolism, we found that low glucose concentration led to a reduction of oxygen consumption rate (OCR), basal and maximal oxidative metabolism, and ATP production only in CAFs, but not in tumour cells. These results suggest that CAFs metabolism impairment was responsible for the decreased invasion process of tumour cells, most likely preventing the release of the pro-migratory factors produced by CAFs. In conclusion, the interplay between CAFs and tumour cells is distinctive depending on the gene involved, and highlights the possibility to inhibit CAF-induced migration by impairing CAFs metabolism, indicating new potential therapeutic scenarios for medical therapy.
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- 2022
12. VEGF-A/VEGFR-1: a painful astrocyte-mediated signaling blocked by the anti-VEGFR-1 mAb D16F7
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Elena Lucarini, Carmen Parisio, Carla Ghelardini, Alessandra Pacini, Serena Boccella, Alessia Vona, Alessandra Toti, L. Di Cesare Mannelli, Grazia Graziani, Paola Failli, Tommaso Mello, Flavia Ricciardi, Sabatino Maione, Pedro Miguel Lacal, and Laura Micheli
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Gene knockdown ,education.field_of_study ,business.industry ,Population ,Neurotoxicity ,Context (language use) ,Pharmacology ,medicine.disease ,Vascular endothelial growth factor A ,medicine.anatomical_structure ,Mediator ,Neuropathic pain ,medicine ,business ,education ,Astrocyte - Abstract
Chemotherapy-induced neuropathic pain is a clinically relevant adverse effect of several anticancer drugs leading to dose reduction or therapy discontinuation. The lack of knowledge about the mechanisms of neuropathy development and pain chronicization makes chemotherapy-induced neuropathic pain treatment an unmet medical need. In this context, the vascular endothelial growth factor A (VEGF-A) has emerged as a neurotoxicity biomarker in a model of chemotherapy-induced neuropathy, and its decrease has been related to pain relief. Aim of this study was to clarify the VEGF-A-dependent pain signaling in the CNS for individuating new targeted therapeutic approaches. In mice, the intrathecal infusion of VEGF-A induced a dose-dependent noxious hypersensitivity mediated by the VEGF receptor 1 (VEGFR-1) as demonstrated by pharmacological and genetic tools. In electrophysiological study, VEGF-A stimulated the spinal nociceptive neurons activity through VEGFR-1. In the dorsal horn of the spinal cord, VEGF-A increased in astrocytes of animals affected by neuropathy suggesting this cell population as a source of the potent pain mediator. Accordingly, the selective knockdown of astrocytic VEGF-A, by shRNAmir, blocked the development of oxaliplatin-induced neuropathic pain. Besides, the anti-VEGFR-1 mAb D16F7 (previously described as anticancer) effectively relieved neuropathic pain induced by chemotherapeutic agents. In conclusion, astrocyte-released VEGF-A is a new player in the complex neuron-glia network that oversees physiological and pathological pain and D16F7 mAb rises as a potent pain killer strategy.
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- 2021
13. Glucocorticoid-induced leucine zipper regulates liver fibrosis by suppressing CCL2-mediated leukocyte recruitment
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Zenobio Viana de Barros, Timofei S. Zatsepin, Carlo Riccardi, Oxana Bereshchenko, Chiara Fiorucci, Graziella Migliorati, Stefano Pagano, Michele Biagioli, Tommaso Mello, Philipp Sergeev, Tatiana Prikazchikova, Musetta Paglialunga, Sara Flamini, Andrea Gagliardi, Stefano Bruscoli, Institute for Molecular Medicine Finland, Helsinki Institute of Life Science HiLIFE, and University of Helsinki
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GILZ EXPRESSION ,Liver Cirrhosis ,Male ,Cancer Research ,Leucine zipper ,CCR2 ,PROMOTES ,Immunology ,NF-KAPPA-B ,Inflammation ,CCL2 ,Article ,TISSUE-DAMAGE ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Mice ,0302 clinical medicine ,T-LYMPHOCYTES ,INFLAMMATION ,Leukocytes ,Medicine ,Gene silencing ,Animals ,Humans ,Receptor ,Chemokine CCL2 ,030304 developmental biology ,Mice, Knockout ,Transcription Factors ,0303 health sciences ,QH573-671 ,business.industry ,HEPATIC RECRUITMENT ,MURINE MODEL ,Cell Biology ,Chronic inflammation ,Hepatic stellate cell activation ,Experimental models of disease ,IMMUNE REGULATION ,030220 oncology & carcinogenesis ,Knockout mouse ,CELLS ,Cancer research ,1182 Biochemistry, cell and molecular biology ,medicine.symptom ,business ,Cytology - Abstract
Liver fibrosis (LF) is a dangerous clinical condition with no available treatment. Inflammation plays a critical role in LF progression. Glucocorticoid-induced leucine zipper (GILZ, encoded in mice by the Tsc22d3 gene) mimics many of the anti-inflammatory effects of glucocorticoids, but its role in LF has not been directly addressed. Here, we found that GILZ deficiency in mice was associated with elevated CCL2 production and pro-inflammatory leukocyte infiltration at the early LF stage, resulting in enhanced LF development. RNA interference-mediated in vivo silencing of the CCL2 receptor CCR2 abolished the increased leukocyte recruitment and the associated hepatic stellate cell activation in the livers of GILZ knockout mice. To highlight the clinical relevance of these findings, we found that TSC22D3 mRNA expression was significantly downregulated and was inversely correlated with that of CCL2 in the liver samples of patients with LF. Altogether, these data demonstrate a protective role of GILZ in LF and uncover the mechanism, which can be targeted therapeutically. Therefore, modulating GILZ expression and its downstream targets represents a novel avenue for pharmacological intervention for treating LF and possibly other liver inflammatory disorders.
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- 2021
14. Lipid Cubic Mesophases Combined with Superparamagnetic Iron Oxide Nanoparticles: A Hybrid Multifunctional Platform with Tunable Magnetic Properties for Nanomedical Applications
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Lorenzo Di Cesare Mannelli, Carla Ghelardini, Marco Mendozza, Tommaso Mello, Claudio Sangregorio, Beatrice Muzzi, Costanza Montis, Lucrezia Caselli, Debora Berti, and Alessandra Toti
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Phase transition ,Materials science ,QH301-705.5 ,Nanoparticle ,Antineoplastic Agents ,Nanotechnology ,Adenocarcinoma ,Article ,Catalysis ,Inorganic Chemistry ,Drug Delivery Systems ,X-Ray Diffraction ,Liquid crystal ,Scattering, Small Angle ,Humans ,lyotropic liquid crystals ,Cubic phases ,Cubosomes ,Drug delivery ,Lyotropic liquid crystals ,Magnetic properties ,Nanoparticles ,SPIONs ,Physical and Theoretical Chemistry ,Biology (General) ,Molecular Biology ,QD1-999 ,magnetic nanoparticles, drug delivery, cubosomes, magnetic fields ,Spectroscopy ,cubosomes ,Magnetic energy ,Small-angle X-ray scattering ,Organic Chemistry ,General Medicine ,cubic phases ,Computer Science Applications ,Drug Liberation ,Chemistry ,phase transition ,Lyotropic liquid crystal ,drug delivery ,Magnetic nanoparticles ,Magnetic Iron Oxide Nanoparticles ,nanoparticles ,magnetic properties ,Hybrid material ,HT29 Cells - Abstract
Hybrid materials composed of Superparamagnetic Iron Oxide Nanoparticles (SPIONs) and lipid self-assemblies possess a considerable applicative potential in the biomedical field, specifically, for drug/nutrients delivery. In recent works we have shown that SPION-doped lipid cubic liquid crystals undergo a cubic-to-hexagonal phase transition under the action of temperature or of an alternating magnetic fields (AMF). This transition triggers the release of drugs embedded in the lipid scaffold or in the water channels. In this contribution, we address this phenomenon in depth, to fully elucidate the structural details and optimize the design of hybrid multifunctional carriers for drug delivery. Combining Small-Angle X-Ray Scattering (SAXS) with a magnetic characteri-zation, we find that in bulk lipid cubic phases the cubic-to-hexagonal transition determines the magnetic response of SPIONs. We then extend the investigation from bulk liquid-crystalline phases to colloidal dispersions, i.e. to lipid/SPIONs nanoparticles with cubic internal structure (“magnetocubosomes”). Through Synchrotron SAXS, we monitored the structural response of magnetocubosomes while exposed to an AMF: the magnetic energy, converted into heat by SPIONs, activates the cubic-to-hexagonal transition, and can thus be used as a remote stimulus to spike drug release “on-demand”. In addition, we show that the AMF-induced phase transition in magnetocubosomes steers the re-alignment of SPIONs into linear string assemblies and connect this effect with the change in their magnetic properties, observed at the bulk level. Finally, we assess the internalization ability and cytotoxicity of magnetocubosomes in vitro on HT29 ade-nocarcinoma cancer cells, in order to test the applicability of these smart carriers in drug delivery applications
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- 2021
15. DNA damage response protein CHK2 regulates metabolism in liver cancer
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Krista Rombouts, Matteo Lulli, Paolo Forte, Francesco Paolo Fanizzi, Laura Gragnani, Antonio Mazzocca, Vinicio Carloni, Laura Del Coco, Tommaso Mello, S. Madiai, Andrea Galli, Caecilia H. C. Sukowati, Lulli, M., Del Coco, L., Mello, T., Sukowati, C., Madiai, S., Gragnani, L., Forte, P., Fanizzi, F. P., Mazzocca, A., Rombouts, K., Galli, A., and Carloni, V.
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0301 basic medicine ,Male ,Cancer Research ,Carcinoma, Hepatocellular ,DNA damage ,Citric Acid Cycle ,Mitosis ,Mitochondrion ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Animals ,Humans ,Glycolysis ,Checkpoint Kinase 2 ,Gene knockdown ,Effector ,Chemistry ,Liver Neoplasms ,Succinates ,G2-M DNA damage checkpoint ,Cell biology ,Mitochondria ,Citric acid cycle ,Mice, Inbred C57BL ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,Metabolome ,Reactive Oxygen Species ,Transcriptome ,DNA Damage - Abstract
Defective mitosis with chromosome missegregation can have a dramatic effect on genome integrity by causing DNA damage, activation of the DNA damage response (DDR), and chromosomal instability. Although this is an energy-dependent process, mechanisms linking DDR to cellular metabolism are unknown. Here we show that checkpoint kinase 2 (CHK2), a central effector of DDR, regulates cellular energy production by affecting glycolysis and mitochondrial functions. Patients with hepatocellular carcinoma (HCC) had increased CHK2 mRNA in blood, which was associated with elevated tricarboxylic acid cycle (TCA) metabolites. CHK2 controlled expression of succinate dehydrogenase (SDH) and intervened with mitochondrial functions. DNA damage and CHK2 promoted SDH activity marked by increased succinate oxidation through the TCA cycle; this was confirmed in a transgenic model of HCC with elevated DNA damage. Mitochondrial analysis identified CHK2-controlled expression of SDH as key in sustaining reactive oxygen species production. Cells with DNA damage and elevated CHK2 relied significantly on glycolysis for ATP production due to dysfunctional mitochondria, which was abolished by CHK2 knockdown. This represents a vulnerability created by the DNA damage response that could be exploited for development of new therapies. Significance: This study uncovers a link between a central effector of DNA damage response, CHK2, and cellular metabolism, revealing potential therapeutic strategies for targeting hepatocellular carcinoma.
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- 2021
16. Activated fibroblasts enhance cancer cell migration by microvesicles-mediated transfer of Galectin-1
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Paolo Cirri, Ilaria Nesi, Alessandra Toti, Elisa Pardella, Alice Santi, Anna Caselli, Paolo De Paoli, Tommaso Mello, and Richard Tomasini
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0301 basic medicine ,Tumor microenvironment ,Stromal cell ,Chemistry ,Cell Biology ,Cancer associated fibroblasts ,Cell migration ,Extracellular vesicles ,Galectin-1 ,Microvesicles ,Biochemistry ,Cell biology ,03 medical and health sciences ,Crosstalk (biology) ,030104 developmental biology ,0302 clinical medicine ,Downregulation and upregulation ,Tumor progression ,030220 oncology & carcinogenesis ,Cancer cell ,Original Research Article ,Molecular Biology - Abstract
Cancer-associated fibroblasts (CAFs) are one of the main components of the stromal compartment in the tumor microenvironment (TME) and the crosstalk between CAFs and cancer cells is essential for tumor progression and aggressiveness. Cancer cells mediate an activation process, converting normal fibroblasts into CAFs, that are characterized by modified expression of many proteins and increased production and release of microvesicles (MVs), extracellular vesicles generated by outwards budding from the cell membrane. Recent evidence underlined that the uptake of CAF-derived MVs changes the overall protein content of tumor cells. In this paper, we demonstrate that tumor activated fibroblasts overexpress Galectin-1 (Gal-1) and consequently release MVs containing increased levels of this protein. The uptake of Gal-1 enriched MVs by tumor cells leads to the upregulation of its intracellular concentration, that strongly affects cancer cell migration, while neither proliferation nor adhesion are altered. Accordingly, tumor cells co-cultured with fibroblasts silenced for Gal-1 have a reduced migratory ability. The present work reveals the key role of an exogenous protein, Gal-1, derived from activated fibroblasts, in cancer progression, and contributes to clarify the importance of MVs-mediated protein trafficking in regulating tumor-stroma crosstalk.
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- 2020
17. Effect of acetylsalicylic acid on inflamed adipose tissue. Insulin resistance and hepatic steatosis in a mouse model of diet-induced obesity
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Tommaso Mello, Simone Camelliti, Lucia Sfondrini, Elisa Martini, Michele Sommariva, Cristiano Rumio, Marinos Kallikourdis, Fabrizio Marcucci, and Claudia Sardi
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0301 basic medicine ,medicine.medical_specialty ,Adipose tissue ,Inflammation ,Diet, High-Fat ,030226 pharmacology & pharmacy ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Insulin resistance ,Weight loss ,Adipocyte ,Internal medicine ,Diabetes mellitus ,medicine ,Animals ,Obesity ,General Pharmacology, Toxicology and Pharmaceutics ,Aspirin ,business.industry ,Anti-Inflammatory Agents, Non-Steroidal ,General Medicine ,medicine.disease ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,Endocrinology ,Treatment Outcome ,chemistry ,Adipose Tissue ,Female ,medicine.symptom ,Steatosis ,Adipocyte hypertrophy ,Insulin Resistance ,business - Abstract
Aims Obesity represents a global health problem. Excessive caloric intake promotes the release of inflammatory mediators by hypertrophic adipocytes and obesity-induced inflammation is now recognized as a risk factor for the development of several diseases, such as cardiovascular diseases, insulin resistance, type-II diabetes, liver steatosis and cancer. Since obesity causes inflammation, we tested the ability of acetylsalicylic acid (ASA), a potent anti-inflammatory drug, in counteracting this inflammatory process and in mitigating obesity-associated health complications. Main Methods Mice were fed with standard (SD) or high fat diet (HFD) for 3 months and then treated with acetylsalicylic acid for the subsequent two months. We then analyzed the metabolic and inflammatory status of their adipose and liver tissue by histological, molecular and biochemical analysis. Key findings Although ASA did not exert any effect on body weight, quantification of adipocyte size revealed that the drug slightly reduced adipocyte hypertrophy, however not sufficient so as to induce weight loss. Most importantly, ASA was able to improve insulin resistance. Gene expression profiles of pro- and anti-inflammatory cytokines as well as the expression of macrophage and lymphocyte markers revealed that HFD led to a marked macrophage accumulation in the adipose tissue and an increase of several pro-inflammatory cytokines, a situation almost completely reverted after ASA administration. In addition, liver steatosis caused by HFD was completely abrogated by ASA treatment. Significance ASA can efficiently ameliorate pathological conditions usually associated with obesity by inhibiting the inflammatory process occurring in the adipose tissue.
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- 2020
18. Testosterone treatment is associated with reduced adipose tissue dysfunction and nonalcoholic fatty liver disease in obese hypogonadal men
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Benedetta Beltrame, C. Corno, L Presenti, Marcello Lucchese, Linda Vignozzi, Andrea Galli, Paolo Comeglio, M C Truglia, Mario Maggi, Farid Saad, Tommaso Mello, Elisa Maseroli, Elena Rapizzi, Sandra Filippi, Francesco Lotti, Ilaria Cellai, Giulia Rastrelli, and Enrico Facchiano
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Adult ,Male ,medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Biopsy ,Adipose tissue ,Intra-Abdominal Fat ,Protective Agents ,Asymptomatic ,chemistry.chemical_compound ,Endocrinology ,Non-alcoholic Fatty Liver Disease ,Internal medicine ,NAFLD ,Nonalcoholic fatty liver disease ,medicine ,Humans ,Testosterone ,Obesity ,Triglyceride ,business.industry ,Hypogonadism ,Middle Aged ,medicine.disease ,Lipid Metabolism ,Mitochondria ,Cross-Sectional Studies ,Treatment Outcome ,chemistry ,Italy ,Liver ,Adipogenesis ,Original Article ,medicine.symptom ,Steatosis ,Insulin Resistance ,business - Abstract
Purpose In both preclinical and clinical settings, testosterone treatment (TTh) of hypogonadism has shown beneficial effects on insulin sensitivity and visceral and liver fat accumulation. This prospective, observational study was aimed at assessing the change in markers of fat and liver functioning in obese men scheduled for bariatric surgery. Methods Hypogonadal patients with consistent symptoms (n = 15) undergoing 27.63 ± 3.64 weeks of TTh were compared to untreated eugonadal (n = 17) or asymptomatic hypogonadal (n = 46) men. A cross-sectional analysis among the different groups was also performed, especially for data derived from liver and fat biopsies. Preadipocytes isolated from adipose tissue biopsies were used to evaluate insulin sensitivity, adipogenic potential and mitochondrial function. NAFLD was evaluated by triglyceride assay and by calculating NAFLD activity score in liver biopsies. Results In TTh-hypogonadal men, histopathological NAFLD activity and steatosis scores, as well as liver triglyceride content were lower than in untreated-hypogonadal men and comparable to eugonadal ones. TTh was also associated with a favorable hepatic expression of lipid handling-related genes. In visceral adipose tissue and preadipocytes, TTh was associated with an increased expression of lipid catabolism and mitochondrial bio-functionality markers. Preadipocytes from TTh men also exhibited a healthier morpho-functional phenotype of mitochondria and higher insulin-sensitivity compared to untreated-hypogonadal ones. Conclusions The present data suggest that TTh in severely obese, hypogonadal individuals induces metabolically healthier preadipocytes, improving insulin sensitivity, mitochondrial functioning and lipid handling. A potentially protective role for testosterone on the progression of NAFLD, improving hepatic steatosis and reducing intrahepatic triglyceride content, was also envisaged. Clinical Trial Registration ClinicalTrials.gov Identifier: NCT02248467, September 25th 2014
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- 2020
19. The G protein-coupled oestrogen receptor, GPER1, mediates direct anti-inflammatory effects of oestrogens in human cholinergic neurones from the nucleus basalis of Meynert
- Author
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Annamaria Morelli, Pasquale Gallina, Giulia Guarnieri, Eglantina Idrizaj, Paolo Comeglio, Roberta Squecco, Mario Maggi, Tommaso Mello, Gabriella B. Vannelli, and Erica Sarchielli
- Subjects
Agonist ,medicine.medical_specialty ,GPR30/GPER1 sublocalisation ,TNFα ,neuroprotection ,oestradiol ,patch-clamp ,medicine.drug_class ,G protein ,Endocrinology, Diabetes and Metabolism ,Anti-Inflammatory Agents ,030209 endocrinology & metabolism ,Inflammation ,Cyclopentanes ,Hippocampal formation ,Nucleus basalis ,Neuroprotection ,Receptors, G-Protein-Coupled ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,Endocrinology ,Internal medicine ,medicine ,Humans ,Estradiol ,Tumor Necrosis Factor-alpha ,Endocrine and Autonomic Systems ,Chemistry ,Estrogens ,Cholinergic Neurons ,Cell biology ,Receptors, Estrogen ,Basal Nucleus of Meynert ,Quinolines ,Cholinergic ,Tumor necrosis factor alpha ,medicine.symptom ,030217 neurology & neurosurgery ,Signal Transduction - Abstract
It has been well established, particularly in animal models, that oestrogens exert neuroprotective effects in brain areas linked to cognitive processes. A key protective role could reside in the capacity of oestrogen to modulate the inflammatory response. However, the direct neuroprotective actions of oestrogens on neurones are complex and remain to be fully clarified. In the present study, we took advantage of a previously characterised primary culture of human cholinergic neurones (hfNBM) from the foetal nucleus basalis of Meynert, which is known to regulate hippocampal and neocortical learning and memory circuits, aiming to investigate the direct effects of oestrogens under inflammatory conditions. Exposure of cells to tumour necrosis factor (TNF)α (10 ng mL-1 ) determined the activation of an inflammatory response, as demonstrated by nuclear factor-kappa B p65 nuclear translocation and cyclooxygenase-2 mRNA expression. These effects were inhibited by treatment with either 17β-oestradiol (E2 ) (10 nmol L-1 ) or G1 (100 nmol L-1 ), the selective agonist of the G protein-coupled oestrogen receptor (GPER1). Interestingly, the GPER1 antagonist G15 abolished the effects of E2 in TNFα-treated cells, whereas the ERα/ERβ inhibitor tamoxifen did not. Electrophysiological measurements in hfNBMs revealed a depolarising effect caused by E2 that was specifically blocked by tamoxifen and not by G15. Conversely, G1 specifically hyperpolarised the cell membrane and also increased both inward and outward currents elicited by a depolarising stimulus, suggesting a modulatory action on hfNBM excitability by GPER1 activation. Interestingly, pretreating cells with TNFα completely blocked the effects of G1 on membrane properties and also significantly reduced GPER1 mRNA expression. In addition, we found a peculiar subcellular localisation of GPER1 to focal adhesion sites that implicates new possible mechanisms of action of GPER1 in the neuronal perception of mechanical stimuli. The results obtained in the present study indicate a modulatory functional role of GPER1 with respect to mediating the oestrogen neuroprotective effect against inflammation in brain cholinergic neurones and, accordingly, may help to identify protective strategies for preventing cognitive impairments.
- Published
- 2020
20. Increased Risk of Liver Cirrhosis during Azathioprine Therapy for Crohn’s Disease
- Author
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Monica Milla, Jenny Roselli, E.N. Lynch, Mirko Tarocchi, Andrea Galli, L. Parisio, Tommaso Innocenti, Giuseppe Macrì, Stefano Milani, and Tommaso Mello
- Subjects
Liver Cirrhosis ,Liver injury ,History ,medicine.medical_specialty ,Crohn's disease ,Cirrhosis ,Polymers and Plastics ,medicine.diagnostic_test ,Side effect ,business.industry ,Case Report ,Azathioprine ,RC799-869 ,Diseases of the digestive system. Gastroenterology ,medicine.disease ,Pancytopenia ,Gastroenterology ,Industrial and Manufacturing Engineering ,Internal medicine ,Liver biopsy ,medicine ,Business and International Management ,business ,Transient elastography ,medicine.drug - Abstract
Azathioprine is a cornerstone of the therapy of Crohn’s disease. Unfortunately, infections and malignancies are relatively common adverse effects related to this drug; however, cirrhosis is exceptionally reported as a side effect. We report the case of a 49-year-old male patient with ileocolonic steno-penetrating Crohn’s disease who developed hepatic cirrhosis while treated with azathioprine. After taking azathioprine for 3 years with regular follow-up, he developed pancytopenia, and liver cirrhosis was diagnosed with ultrasound, abdomen computed tomography scan, transient elastography, and liver biopsy. As all other causes of liver damage were excluded, azathioprine was believed to be the cause of liver injury and therefore was interrupted.
- Published
- 2020
21. Stevens–Johnson Syndrome and Herpes Simplex Type 1 Infection during Adalimumab Therapy for Crohn’s Disease
- Author
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Giuseppe Macrì, Tommaso Mello, Maria Rosa Biagini, Tommaso Innocenti, L. Parisio, Jenny Roselli, Pasquale Apolito, Stefano Milani, E.N. Lynch, Mirko Tarocchi, Andrea Galli, and Monica Milla
- Subjects
musculoskeletal diseases ,History ,medicine.medical_specialty ,Polymers and Plastics ,Mucocutaneous zone ,Case Report ,Crohn's Disease ,Disease ,RC799-869 ,medicine.disease_cause ,Industrial and Manufacturing Engineering ,Virus ,030207 dermatology & venereal diseases ,03 medical and health sciences ,0302 clinical medicine ,0502 economics and business ,Adalimumab ,medicine ,Business and International Management ,skin and connective tissue diseases ,Crohn's disease ,business.industry ,Mortality rate ,05 social sciences ,Diseases of the digestive system. Gastroenterology ,medicine.disease ,Dermatology ,humanities ,stomatognathic diseases ,Herpes simplex virus ,050211 marketing ,business ,Adverse drug reaction ,medicine.drug - Abstract
Stevens–Johnson syndrome (SJS) is a severe mucocutaneous adverse drug reaction with a relatively high mortality rate. SJS is described during herpes simplex virus type 1 (HSV1) infection and, rarely, even during adalimumab therapy. We report the case of a patient with Crohn’s disease who developed SJS during an HSV1 infection and a contemporaneous anti-TNFα therapy with adalimumab. Remission was achieved with suspension of adalimumab and high doses of intravenous steroids and antivirals. Patients with HSV1 infection and on adalimumab therapy have a combined risk of SJS and should be monitored closely.
- Published
- 2020
22. Selective HCN1 block as a strategy to control oxaliplatin-induced neuropathy
- Author
-
Annunziatina Laurino, Laura Micheli, Laura Sartiani, Elisabetta Cerbai, Carla Ghelardini, Maria Novella Romanelli, Guido Mannaioni, L. Di Cesare Mannelli, Valentina Spinelli, Alessio Masi, Francesco Resta, and Tommaso Mello
- Subjects
Male ,0301 basic medicine ,Potassium Channels ,Organoplatinum Compounds ,HCN channels ,HCN1 ,Ih current ,Neuropathic pain ,Oxaliplatin ,Pharmacology ,Cellular and Molecular Neuroscience ,Antineoplastic Agents ,03 medical and health sciences ,0302 clinical medicine ,Downregulation and upregulation ,Heart Rate ,Ganglia, Spinal ,Bradycardia ,Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ,Potassium Channel Blockers ,medicine ,Animals ,Rats, Wistar ,Adverse effect ,Cells, Cultured ,Analgesics ,business.industry ,Nociceptors ,Peripheral Nervous System Diseases ,Cardiac action potential ,Benzazepines ,medicine.disease ,030104 developmental biology ,Peripheral neuropathy ,Allodynia ,Tolerability ,Hyperalgesia ,Neuralgia ,medicine.symptom ,business ,030217 neurology & neurosurgery - Abstract
Chemotherapy-Induced Peripheral Neuropathy (CIPN) is the most frequent adverse effect of pharmacological cancer treatments. The occurrence of neuropathy prevents the administration of fully-effective drug regimen, affects negatively the quality of life of patients, and may lead to therapy discontinuation. CIPN is currently treated with anticonvulsants, antidepressants, opioids and non-opioid analgesics, all of which are flawed by insufficient anti-hyperalgesic efficacy or addictive potential. Understandably, developing new drugs targeting CIPN-specific pathogenic mechanisms would dramatically improve efficacy and tolerability of anti-neuropathic therapies. Neuropathies are associated to aberrant excitability of DRG neurons due to the alteration in the expression or function of a variety of ion channels. In this regard, Hyperpolarization-activated Cyclic Nucleotide-gated (HCN) channels are overexpressed in inflammatory and neuropathic pain states, and HCN blockers have been shown to reduce neuronal excitability and to ameliorate painful states in animal models. However, HCN channels are critical in cardiac action potential, and HCN blockers used so far in pre-clinical models do not discriminate between cardiac and non-cardiac HCN isoforms. In this work, we show an HCN current gain of function in DRG neurons from oxaliplatin-treated rats. Biochemically, we observed a downregulation of HCN2 expression and an upregulation of the HCN regulatory beta-subunit MirP1. Finally, we report the efficacy of the selective HCN1 inhibitor MEL57A in reducing hyperalgesia and allodynia in oxaliplatin-treated rats without cardiac effects. In conclusion, this study strengthens the evidence for a disease-specific role of HCN1 in CIPN, and proposes HCN1-selective inhibitors as new-generation pain medications with the desired efficacy and safety profile.
- Published
- 2018
23. OC.10.8 DISHEVELLED-ASSOCIATED ANTAGONIST OF B-CATENIN 2 (DACT2) IN PANCREATIC DUCTAL ADENOCARCINOMA
- Author
-
Tommaso Mello, L. Picariello, I. Simeone, Elisabetta Ceni, Cecilia Grappone, Andrea Galli, Stefano Milani, Simone Polvani, A. Guida, Gabriele Dragoni, and Mirko Tarocchi
- Subjects
chemistry.chemical_classification ,Pancreatic ductal adenocarcinoma ,Hepatology ,chemistry ,business.industry ,Catenin ,Gastroenterology ,Antagonist ,Cancer research ,Medicine ,business ,Dishevelled - Published
- 2021
24. DNA damage response protein checkpoint kinase 2 (CHK2) links chromosomal instability to cellular metabolism in hepatocellular carcinoma (HCC)
- Author
-
Matteo Lulli, Veronica Ghini, Tommaso Mello, Caecilia Sukowati, Antonio Mazzocca, Paola Turano, Krista Rombouts, Andrea Galli, and Vinicio Carloni
- Subjects
Hepatology - Published
- 2020
25. The effects of testosterone treatment on fat tissue dysfunction and nonalcoholic fatty liver disease in obese men undergoing bariatric surgery
- Author
-
Tommaso Mello, Benedetta Beltrame, Marcello Lucchese, Andrea Galli, Paolo Comeglio, Ilaria Cellai, Linda Vignozzi, Giulia Rastrelli, Mario Maggi, C. Corno, Sandra Filippi, Elisa Maseroli, and Enrico Facchiano
- Subjects
medicine.medical_specialty ,business.industry ,Testosterone treatment ,Internal medicine ,Nonalcoholic fatty liver disease ,Medicine ,Adipose tissue ,business ,medicine.disease ,Gastroenterology - Published
- 2019
26. Mito-Nuclear Communication in Hepatocellular Carcinoma Metabolic Rewiring
- Author
-
Tommaso Mello, I. Simeone, and Andrea Galli
- Subjects
Carcinoma, Hepatocellular ,HCC ,HIF ,NRF ,PGC-1 ,PPAR ,SIRT ,liver ,metabolism ,mitochondria ,Peroxisome proliferator-activated receptor ,Review ,Drug resistance ,Mitochondrion ,Chronic liver disease ,Epigenesis, Genetic ,medicine ,Animals ,Humans ,Neoplastic transformation ,lcsh:QH301-705.5 ,Cell Nucleus ,chemistry.chemical_classification ,business.industry ,Liver Neoplasms ,General Medicine ,medicine.disease ,Crosstalk (biology) ,chemistry ,lcsh:Biology (General) ,Hepatocellular carcinoma ,Cancer cell ,Cancer research ,business ,Signal Transduction - Abstract
As the main metabolic and detoxification organ, the liver constantly adapts its activity to fulfill the energy requirements of the whole body. Despite the remarkable adaptive capacity of the liver, prolonged exposure to noxious stimuli such as alcohol, viruses and metabolic disorders results in the development of chronic liver disease that can progress to hepatocellular carcinoma (HCC), which is currently the second leading cause of cancer-related death worldwide. Metabolic rewiring is a common feature of cancers, including HCC. Altered mito-nuclear communication is emerging as a driving force in the metabolic reprogramming of cancer cells, affecting all aspects of cancer biology from neoplastic transformation to acquired drug resistance. Here, we explore relevant aspects (and discuss recent findings) of mito-nuclear crosstalk in the metabolic reprogramming of hepatocellular carcinoma.
- Published
- 2019
27. A mitochondrial role of RuvBL1 ATPase in the regulation of HCC cell metabolism
- Author
-
D. Bani, M. Materozzi, I. Simeone, Simone Polvani, A. Guida, Andrea Galli, Elisabetta Ceni, M. Parri, and Tommaso Mello
- Subjects
Cell metabolism ,Hepatology ,biology ,business.industry ,ATPase ,Gastroenterology ,biology.protein ,Medicine ,business ,Cell biology - Published
- 2021
28. Non-small-bowel lesions identification by capsule endoscopy: A single centre retrospective study
- Author
-
Andrea Galli, Jenny Roselli, Stefano Milani, Gabriele Dragoni, Giuseppe Macrì, Tommaso Mello, and Tommaso Innocenti
- Subjects
medicine.medical_specialty ,Colonoscopy ,Capsule Endoscopy ,Endoscopy, Gastrointestinal ,law.invention ,Lesion ,03 medical and health sciences ,Ileocecal valve ,0302 clinical medicine ,Capsule endoscopy ,law ,Gastroscopy ,medicine ,Humans ,Retrospective Studies ,Hepatology ,medicine.diagnostic_test ,business.industry ,Incidence (epidemiology) ,Gastroenterology ,Capsule ,Retrospective cohort study ,Endoscopy ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,030211 gastroenterology & hepatology ,Radiology ,medicine.symptom ,Gastrointestinal Hemorrhage ,business - Abstract
Summary Background Capsule endoscopy has been considered the first-line approach for the investigation of obscure gastro-intestinal bleeding since its approval in 2001. Our study aims to evaluate the diagnostic yield of capsule endoscopy in the investigation of this condition. We also analyse the incidence of non-small-bowel lesions missed after conventional endoscopy and later detected by capsule endoscopy in patients with suspected obscure bleeding. Methods A total of 290 patients with negative conventional endoscopy referred to our centre to undergo a capsule endoscopy examination for the investigation of obscure gastro-intestinal bleeding. We considered as non-small-bowel lesions those outside the tract between the second duodenal portion and the ileocecal valve. We also looked for actively bleeding lesions at the time of the exam. Results Intestinal preparation was good, adequate or poor in 74.1%, 8.4%, and 17.5% of the tests, respectively. Caecum was reached in 92.4%. Capsule retention occurred in 0.7%. Mean small bowel transit time was 5 hours and 13 minutes. Diagnostic yield was 73.8%. An actively bleeding lesion was noticed in 39.3% of positive tests. Capsule endoscopy revealed clinically significant non-small-bowel lesions missed at gastroscopy or colonoscopy in 30.3% of patients, 43.2% of which were bleeding. Conclusions Capsule endoscopy has high diagnostic yield and safety in the investigation of obscure gastro-intestinal bleedings. Given the high percentage of non-small-bowel lesions detected, it may be appropriate to consider an endoscopic second look before performing a capsule endoscopy study.
- Published
- 2021
29. Tadalafil reduces visceral adipose tissue accumulation by promoting preadipocytes differentiation towards a metabolically healthy phenotype: Studies in rabbits
- Author
-
Francesca Corcetto, Paolo Comeglio, Ilaria Cellai, Tommaso Mello, Andrea Lenzi, Elena Maneschi, Giulia Salvatore, Annamaria Morelli, Gabriella B. Vannelli, Andrea Galli, Linda Vignozzi, Daniele Guasti, Daniele Bani, Erica Sarchielli, Benedetta Mazzanti, C. Corno, Mario Maggi, Edoardo Mannucci, Sandra Filippi, Davide Francomano, and Antonio Aversa
- Subjects
Male ,0301 basic medicine ,medicine.medical_specialty ,Intra-Abdominal Fat ,medicine.medical_treatment ,Adipose tissue ,030209 endocrinology & metabolism ,Mitochondrion ,Biology ,Diet, High-Fat ,Biochemistry ,Tadalafil ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Insulin resistance ,Adipose Tissue, Brown ,Internal medicine ,medicine ,Animals ,Molecular Biology ,Uncoupling Protein 1 ,Metabolic Syndrome ,Insulin ,Hypertriglyceridemia ,nutritional and metabolic diseases ,Cell Differentiation ,medicine.disease ,Thermogenin ,Mitochondria ,Disease Models, Animal ,030104 developmental biology ,Gene Expression Regulation ,Rabbits ,Metabolic syndrome - Abstract
Development of metabolically healthy adipocytes within dysfunctional adipose tissue may represent an attractive way to counteract metabolic syndrome (MetS). In an experimental animal model of high fat diet (HFD)-induced MetS, in vivo, long- and short-term tadalafil treatments were able to reduce visceral adipose tissue (VAT) accumulation and hypertriglyceridemia, and to induce the expression in VAT of the brown fat-specific marker, uncoupling protein 1 (UCP1). VAT preadipocytes (PAD), isolated from the tadalafil-treated HFD rabbits, showed: i) a multilocular morphology; ii) an increased expression of brown fat-specific genes (such as UCP1 and CIDEA); iii) improved mitochondrial structure and dynamic and reduced superoxide production; iv) improved insulin sensitivity. Similar effects were obtained after in vitro tadalafil treatment in HFD rPAD. In conclusion, tadalafil counteracted HFD-associated VAT alterations, by restoring insulin-sensitivity and prompting preadipocytes differentiation towards a metabolically healthy phenotype.
- Published
- 2016
30. Oxidative Stress in the Healthy and Wounded Hepatocyte: A Cellular Organelles Perspective
- Author
-
Andrea Galli, Elisabetta Ceni, Tommaso Mello, and F. Zanieri
- Subjects
0301 basic medicine ,Aging ,Review Article ,Oxidative phosphorylation ,Biology ,medicine.disease_cause ,Models, Biological ,Biochemistry ,03 medical and health sciences ,Organelle ,medicine ,Animals ,Humans ,lcsh:QH573-671 ,Cellular compartment ,Organelles ,chemistry.chemical_classification ,Reactive oxygen species ,lcsh:Cytology ,Cell migration ,Cell Biology ,General Medicine ,Cell biology ,Oxidative Stress ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,oxidative stress, ROS, Cellular Organelles ,Hepatocyte ,Hepatocytes ,Reactive Oxygen Species ,Oxidative stress ,Homeostasis - Abstract
Accurate control of the cell redox state is mandatory for maintaining the structural integrity and physiological functions. This control is achieved both by a fine-tuned balance between prooxidant and anti-oxidant molecules and by spatial and temporal confinement of the oxidative species. The diverse cellular compartments each, although structurally and functionally related, actively maintain their own redox balance, which is necessary to fulfill specialized tasks. Many fundamental cellular processes such as insulin signaling, cell proliferation and differentiation and cell migration and adhesion, rely on localized changes in the redox state of signal transducers, which is mainly mediated by hydrogen peroxide (H2O2). Therefore, oxidative stress can also occur long before direct structural damage to cellular components, by disruption of the redox circuits that regulate the cellular organelles homeostasis. The hepatocyte is a systemic hub integrating the whole body metabolic demand, iron homeostasis and detoxification processes, all of which are redox-regulated processes. Imbalance of the hepatocyte’s organelles redox homeostasis underlies virtually any liver disease and is a field of intense research activity. This review recapitulates the evolving concept of oxidative stress in the diverse cellular compartments, highlighting the principle mechanisms of oxidative stress occurring in the healthy and wounded hepatocyte.
- Published
- 2016
31. T02.02.6 NON-SMALL-BOWEL LESIONS IDENTIFICATION BY CAPSULE ENDOSCOPY: A SINGLE CENTRE RETROSPECTIVE STUDY
- Author
-
Tommaso Mello, Jenny Roselli, Tommaso Innocenti, Stefano Milani, Gabriele Dragoni, G. Macrì, and Andrea Galli
- Subjects
Single centre ,medicine.medical_specialty ,Hepatology ,business.industry ,Capsule endoscopy ,law ,Gastroenterology ,Medicine ,Identification (biology) ,Retrospective cohort study ,Radiology ,business ,law.invention - Published
- 2020
32. Oxidative Stress and DNA Damage in Chronic Disease and Environmental Studies
- Author
-
Tommaso Mello, Valentina Russo, Marco Peluso, and Andrea Galli
- Subjects
0301 basic medicine ,DNA damage ,medicine.disease_cause ,Catalysis ,lcsh:Chemistry ,Inorganic Chemistry ,03 medical and health sciences ,0302 clinical medicine ,Occupational Exposure ,medicine ,Humans ,Physical and Theoretical Chemistry ,lcsh:QH301-705.5 ,Molecular Biology ,Spectroscopy ,business.industry ,Organic Chemistry ,General Medicine ,Computer Science Applications ,Oxidative Stress ,Editorial ,n/a ,030104 developmental biology ,Chronic disease ,lcsh:Biology (General) ,lcsh:QD1-999 ,030220 oncology & carcinogenesis ,Chronic Disease ,Immunology ,business ,Oxidative stress ,DNA Damage ,Environmental Monitoring - Abstract
Humans are continually exposed to a large number of environmental carcinogens [...]
- Published
- 2020
33. Effects of Cadmium on ZO-1 Tight Junction Integrity of the Blood Brain Barrier
- Author
-
Carla Ghelardini, Cristina Gonnelli, Tommaso Mello, Claudio Nicoletti, Alessandra Pacini, Mario Maresca, Luigia Pazzagli, Matteo Becatti, Lorenzo Di Cesare Mannelli, Donatello Carrino, Ferdinando Paternostro, Jacopo Junio Valerio Branca, Ilaria Colzi, Massimo Gulisano, and Gabriele Morucci
- Subjects
cadmium ,blood brain barrier ,Vimentin ,Cadmium chloride ,Blood–brain barrier ,RBE4 cell line ,Article ,Catalysis ,Cell Line ,Tight Junctions ,ATP extrusion ,Inorganic Chemistry ,chemistry.chemical_compound ,medicine ,Animals ,ROS production ,Physical and Theoretical Chemistry ,caspase-3 activation ,Molecular Biology ,ZO-1 ,Spectroscopy ,biology ,Tight junction ,Endoplasmic reticulum ,Organic Chemistry ,Neurodegeneration ,Endothelial Cells ,General Medicine ,Endoplasmic Reticulum Stress ,medicine.disease ,Actins ,Rats ,Computer Science Applications ,Cell biology ,Endothelial stem cell ,medicine.anatomical_structure ,chemistry ,Blood-Brain Barrier ,ER stress ,Zonula Occludens-1 Protein ,biology.protein ,Unfolded protein response ,Reactive Oxygen Species - Abstract
Cadmium (Cd) is a highly toxic environmental pollutant released from the smelting and refining of metals and cigarette smoking. Oral exposure to cadmium may result in adverse effects on a number of tissues, including the central nervous system (CNS). In fact, its toxicity has been related to neurological disorders, as well as neurodegenerative diseases such as Alzheimer&rsquo, s and Parkinson&rsquo, s diseases. Under normal conditions, Cd barely reaches the brain in adults because of the presence of the blood&ndash, brain barrier (BBB), however, it has been demonstrated that Cd-dependent BBB alteration contributes to pathogenesis of neurodegeneration. However, the mechanism underlying Cd-dependent BBB alteration remain obscure. Here, we investigated the signaling pathway of Cd-induced tight junction (TJ), F-actin, and vimentin protein disassembly in a rat brain endothelial cell line (RBE4). RBE4 cells treated with 10 &mu, M cadmium chloride (CdCl2) showed a dose- and time-dependent significant increase in reactive oxygen species (ROS) production. This phenomenon was coincident with the alteration of the TJ zonula occludens-1 (ZO-1), F-actin, and vimentin proteins. The Cd-dependent ROS increase elicited the upregulation of GRP78 expression levels, a chaperone involved in endoplasmic reticulum (ER) stress that induces caspase-3 activation. Further signal profiling by the pannexin-1 (PANX1) specific inhibitor 10Panx revealed a PANX1-independent increase in ATP spillage in Cd-treated endothelial cells. Our results point out that a ROS-dependent ER stress-mediated signaling pathway involving caspase-3 activation and ATP release is behind the BBB morphological alterations induced by Cd.
- Published
- 2019
34. INT-767 prevents NASH and promotes visceral fat brown adipogenesis and mitochondrial function
- Author
-
Linda Vignozzi, Erica Sarchielli, Tommaso Mello, Paolo Comeglio, Andrea Galli, Elena Maneschi, Gabriella B. Vannelli, Daniele Bani, Mario Maggi, Luciano Adorini, Francesca Corcetto, Daniele Guasti, Elena Rapizzi, Annamaria Morelli, Ilaria Cellai, Sandra Filippi, and C. Corno
- Subjects
0301 basic medicine ,Adult ,Male ,medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Adipose tissue ,Receptors, Cytoplasmic and Nuclear ,Intra-Abdominal Fat ,Bile Acids and Salts ,03 medical and health sciences ,Endocrinology ,Insulin resistance ,Adipose Tissue, Brown ,Non-alcoholic Fatty Liver Disease ,Internal medicine ,medicine ,Adipocytes ,Animals ,Humans ,Obesity ,Cells, Cultured ,Aged ,Metabolic Syndrome ,Adipogenesis ,biology ,Chemistry ,Lipid metabolism ,Cell Differentiation ,Middle Aged ,medicine.disease ,G protein-coupled bile acid receptor ,Mitochondria ,INT-767 ,insulin resistance ,metabolic syndrome ,mitochondrial metabolism ,preadipocytes ,Insulin receptor ,Disease Models, Animal ,030104 developmental biology ,Mitochondrial biogenesis ,Liver ,biology.protein ,Farnesoid X receptor ,Rabbits - Abstract
The bile acid receptors, farnesoid X receptor (FXR) and Takeda G-protein-coupled receptor 5 (TGR5), regulate multiple pathways, including glucose and lipid metabolism. In a rabbit model of high-fat diet (HFD)-induced metabolic syndrome, long-term treatment with the dual FXR/TGR5 agonist INT-767 reduces visceral adipose tissue accumulation, hypercholesterolemia and nonalcoholic steatohepatitis. INT-767 significantly improves the hallmarks of insulin resistance in visceral adipose tissue (VAT) and induces mitochondrial and brown fat-specific markers. VAT preadipocytes isolated from INT-767-treated rabbits, compared to preadipocytes from HFD, show increased mRNA expression of brown adipogenesis markers. In addition, INT-767 induces improved mitochondrial ultrastructure and dynamic, reduced superoxide production and improved insulin signaling and lipid handling in preadipocytes. Bothin vivoandin vitrotreatments with INT-767 counteract, in preadipocytes, the HFD-induced alterations by upregulating genes related to mitochondrial biogenesis and function. In preadipocytes, INT-767 behaves mainly as a TGR5 agonist, directly activating dose dependently the cAMP/PKA pathway. However,in vitroexperiments also suggest that FXR activation by INT-767 contributes to the insulin signaling improvement. INT-767 treatment counteracts HFD-induced liver histological alterations and normalizes the increased pro-inflammatory genes. INT-767 also induces a significant reduction of fatty acid synthesis and fibrosis markers, while increasing lipid handling, insulin signaling and mitochondrial markers. In conclusion, INT-767 significantly counteracts HFD-induced liver and fat alterations, restoring insulin sensitivity and prompting preadipocytes differentiation toward a metabolically healthy phenotype.
- Published
- 2018
35. Hepatocyte-specific deletion of RuvBL1 leads to chronic liver damage and regeneration, enhancing HCC development
- Author
-
Oxana Bereshchenko, Andrea Galli, M. Materozzi, Mirko Tarocchi, F. Zanieri, Elisabetta Ceni, Simone Polvani, and Tommaso Mello
- Subjects
medicine.anatomical_structure ,Hepatology ,Hepatocyte ,Regeneration (biology) ,medicine ,Cancer research ,Liver damage ,Biology - Published
- 2018
36. Inhibition of spinal ERK1/2–c-JUN signaling pathway counteracts the development of low doses morphine-induced hyperalgesia
- Author
-
Maria Domenica Sanna, Nicoletta Galeotti, Tommaso Mello, and Carla Ghelardini
- Subjects
Male ,MAPK/ERK pathway ,MAP Kinase Signaling System ,Narcotic Antagonists ,Pharmacology ,Mice ,Animals ,Medicine ,Phosphorylation ,Extracellular Signal-Regulated MAP Kinases ,Protein Kinase Inhibitors ,Sensitization ,Flavonoids ,Behavior, Animal ,Morphine ,Naloxone ,business.industry ,Kinase ,c-jun ,JNK Mitogen-Activated Protein Kinases ,Posterior Horn Cells ,medicine.anatomical_structure ,Hyperalgesia ,Signal transduction ,medicine.symptom ,business ,Signal Transduction ,medicine.drug - Abstract
Morphine-induced hyperalgesia is a pharmacological phenomenon often hindering its prolonged applications in the clinic. It has been shown that systemic administration of morphine induced a hyperalgesic response at an extremely low dose. Extracellular signal-regulated kinase (ERK) pathway contributes to pain sensitization, and its phosphorylation under pain conditions results in the induction and maintenance of pain hypersensitivity. The present study was designed to determine whether low dose morphine treatment in mice could influence the spinal activity of ERK. The data showed that morphine (1 µg/kg) induced a marked increase in ERK phosphorylation. Intrathecal pre-treatment with a selective mitogen-activated and extracellular signal-regulated kinase (MEK) inhibitor PD98059, attenuated morphine-associated thermal hyperalgesia. Morphine exposure increased phosphorylation of c-JUN, that was prevented by the inhibition of ERK pathway. In addition, double immunofluorescence studies revealed that, p-ERK and p-c-JUN are localized on neurons of the spinal dorsal horn expressing µ receptors. These data suggest that ERK contributes to the morphine-induced hyperalgesia by regulating the activation of c-JUN.
- Published
- 2015
37. The RNA-binding protein HuD promotes spinal GAP43 overexpression in antiretroviral-induced neuropathy
- Author
-
Alessandro Quattrone, Maria Domenica Sanna, Carla Ghelardini, Tommaso Mello, and Nicoletta Galeotti
- Subjects
Male ,Time Factors ,Pain ,Oligodeoxyribonucleotides, Antisense ,Mice ,GAP-43 Protein ,Developmental Neuroscience ,Animals ,Gene silencing ,Medicine ,Gap-43 protein ,Cells, Cultured ,Protein Kinase C ,Protein kinase C ,Neurons ,ATF3 ,biology ,Zalcitabine ,Activator (genetics) ,Kinase ,business.industry ,Peripheral Nervous System Diseases ,Neuroregeneration ,Disease Models, Animal ,Anti-Retroviral Agents ,ELAV Proteins ,Gene Expression Regulation ,Spinal Cord ,Neurology ,Hyperalgesia ,Phosphopyruvate Hydratase ,Immunology ,Neuropathic pain ,biology.protein ,Cancer research ,business - Abstract
Nucleoside reverse transcriptase inhibitors (NRTIs) are known to produce painful neuropathies and to enhance states of pain hypersensitivity produced by HIV-1 infection in patients with AIDS leading to discontinuation of antiretroviral therapy, thus limiting viral suppression strategies. The mechanisms by which NRTIs contribute to the development of neuropathic pain are not known. In the current study, we tested the hypothesis that HuD, an RNA binding protein known to be an essential promoter of neuronal differentiation and survival, might be involved in the response to NRTI-induced neuropathy. Antiretroviral neuropathy was induced by a single intraperitoneal administration of 2′,3′-dideoxycytidine (ddC) in mice. HuD was physiologically expressed in the cytoplasm of the soma and in axons of neurons within DRG and spinal cord and was considerably overexpressed following ddC treatment. ddC up-regulated spinal GAP43 protein, a marker of neuroregeneration, and this increase was counteracted by HuD silencing. GAP43 and HuD colocalize in DRG and spinal dorsal horn (SDH) axons and administration of an anti-GAP43 antibody aggravated the ddC-induced axonal damage. The administration of a protein kinase C (PKC) inhibitor or the PKCγ silencing prevented both HuD and GAP43 increased expression. Conversely, treatment with the PKC activator PDBu potentiated HuD and GAP43 overexpression, demonstrating the presence of a spinal PKC-dependent HuD–GAP43 pathway activated by ddC. These results indicated that HuD recruitment and GAP43 protein increase are mechanistically linked events involved in the response to antiretroviral-induced neurodegenerative processes.
- Published
- 2014
38. The microenvironment induces collective migration in SDHB-silenced mouse pheochromocytoma spheroids
- Author
-
Massimo Mannelli, Graeme Eisenhofer, Tommaso Mello, Susan Richter, Serena Martinelli, Letizia Canu, Elena Rapizzi, Vanessa D’Antongiovanni, Paolo Romagnoli, Daniele Guasti, and Karel Pacak
- Subjects
0301 basic medicine ,Cancer Research ,medicine.medical_specialty ,SDHB ,Endocrinology, Diabetes and Metabolism ,Adrenal Gland Neoplasms ,spheroids ,Pheochromocytoma ,Article ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Endocrinology ,Germline mutation ,Cell Movement ,Internal medicine ,medicine ,Tumor Microenvironment ,Gene silencing ,Animals ,Humans ,pheochromocytoma/ paraganglioma ,Tumor microenvironment ,biology ,Chemistry ,Wild type ,Spheroid ,biology.organism_classification ,Diabetes and Metabolism ,Succinate Dehydrogenase ,tumor migration ,Pheos ,030104 developmental biology ,Oncology ,Cell culture ,030220 oncology & carcinogenesis ,embryonic structures ,Cancer research ,Female ,tumor microenvironment - Abstract
Pheochromocytomas (Pheos) and paragangliomas (PGLs) are neuroendocrine tumors. Approximately 30–40% of Pheos/PGLs are due to germline mutations in one of the susceptibility genes, including those encoding the succinate dehydrogenase subunits A-D (SDHA-D). Up to 2/3 of patients affected bySDHBmutated Pheo/PGL develop metastatic disease with no successful cure at present. Here, for the first time, we evaluated the effects ofSDHBsilencing in a three dimension (3D) culture using spheroids of a mouse Pheo cell line silenced or not (wild type/wt/control) for the SDHB subunit. We investigated the role of the microenvironment on spheroid growth and migration/invasion by co-culturingSDHB-silenced or wt spheroids with primary cancer-activated fibroblasts (CAFs). When spheroids were co-cultured with fibroblasts,SDHB-silenced cells showed a significant increase in matrigel invasion as demonstrated by the computation of the migratory areas (P SDHB-silenced spheroids moved collectively, unlike the cells of wt spheroids that moved individually. Additionally,SDHB-silenced spheroids developed long filamentous formations along which clusters of cells migrated far away from the spheroid, whereas these structures were not present in wt spheroids. We found that lactate, largely secreted by CAFs, plays a specific role in promoting migration only ofSDHB-silenced cells. In this study, we demonstrated thatSDHBsilencingper seincreases tumor cell migration/invasion and that microenvironment, as represented by CAFs, plays a pivotal role in enhancing collective migration/invasion in PheoSDHB-silenced tumor cells, suggesting their role in increasing the tumor metastasizing potential.
- Published
- 2017
39. Activation of ERK/CREB pathway in noradrenergic neurons contributes to hypernociceptive phenotype in H4 receptor knockout mice after nerve injury
- Author
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Nicoletta Galeotti, Emanuela Masini, Tommaso Mello, and Maria Domenica Sanna
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0301 basic medicine ,MAPK/ERK pathway ,Adrenergic Neurons ,Pain Threshold ,SNi ,medicine.medical_specialty ,Sensory Receptor Cells ,MAP Kinase Signaling System ,Dopamine beta-Hydroxylase ,CREB ,Piperazines ,Histamine Agonists ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Mice ,0302 clinical medicine ,Internal medicine ,Ganglia, Spinal ,medicine ,Animals ,Phosphorylation ,Cyclic AMP Response Element-Binding Protein ,Receptors, Histamine H4 ,Pharmacology ,Mice, Knockout ,ATF3 ,Activating Transcription Factor 3 ,biology ,Chemistry ,Wild type ,Nerve injury ,Spinal cord ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,Endocrinology ,medicine.anatomical_structure ,Phenotype ,Pyrimidines ,Hyperalgesia ,Neuropathic pain ,biology.protein ,Neuralgia ,medicine.symptom ,Neuroscience ,030217 neurology & neurosurgery - Abstract
G-protein coupled receptor H4 (H4R) is a histamine receptor subtype that is involved in a condition of pathological chronic pain, but its pathophysiological function is unknown. Here, we investigate the role of H4R in a model of traumatic nerve injury. H4R knockout (H4R-/-) mice exposed to spared nerve injury (SNI) developed a more prominent mechanical and thermal hypersensitivity than wild type mice. Western blotting and immunofluorescence were used to characterize the cellular mechanisms. Nerve injury increased phosphorylated pERK MAPK expression in the spinal cord that was further promoted in H4R-/- genotype. Additionally, the increase in the phosphorylated cAMP response element-binding protein (CREB) was significantly enhanced in neuropathic H4R-/- mice. In the same way, after SNI a remarkable increase of dopamine beta-hydroxylase (DβH) immunoreactive neurons was detected in spinal cord of H4R-/- mice. The number of injured DRG neurons after SNI of H4R-/- mice, identified by activating transcription factor 3 (ATF3) staining was comparable to that of wild type littermates. Similarly the density of intraepidermal nerve fibres in plantar skin after SNI was reduced with the same degree in H4R-/- mice and with wild type mice. We conclude that the phenotype of H4R-/- mice leads to increased neuropathic pain hypersensitivity promoting an overactivation of spinal ERK-CREB pathway in DβH expressing neurons without modifying the innervation of the hind paw skin and integrity of the primary sensory neurons. In summary, our results provide H4R as a potential new target for the clinical management of chronic neuropathic pain conditions.
- Published
- 2017
40. Magnetic Hyperthermia and Oxidative Damage to DNA of Human Hepatocarcinoma Cells
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Andrea Galli, Elisabetta Ceni, Simone Polvani, Costanza Ravagli, Filippo Cellai, Tommaso Mello, Roger W. Giese, Saer Doumett, Giovanni Baldi, Marco Peluso, Jessica Viti, and Armelle Munnia
- Subjects
0301 basic medicine ,Carcinoma, Hepatocellular ,DNA damage ,M1dG ,8-oxodG ,Oxidative phosphorylation ,medicine.disease_cause ,Article ,Catalysis ,Inorganic Chemistry ,Lipid peroxidation ,lcsh:Chemistry ,DNA Adducts ,03 medical and health sciences ,chemistry.chemical_compound ,Human hepatocarcinoma cells ,Magnetic therapy ,Nanotoxicity ,Molecular Biology ,Computer Science Applications1707 Computer Vision and Pattern Recognition ,Spectroscopy ,Physical and Theoretical Chemistry ,Organic Chemistry ,medicine ,Humans ,Deoxyguanosine ,Magnetite Nanoparticles ,lcsh:QH301-705.5 ,magnetic therapy ,nanotoxicity ,human hepatocarcinoma cells ,Chemistry ,Liver Neoplasms ,Hep G2 Cells ,Hyperthermia, Induced ,General Medicine ,Molecular biology ,Computer Science Applications ,Oxidative Stress ,030104 developmental biology ,Magnetic hyperthermia ,Biochemistry ,lcsh:Biology (General) ,lcsh:QD1-999 ,Nanotoxicology ,Cancer cell ,Lipid Peroxidation ,Oxidative stress ,DNA Damage - Abstract
Nanotechnology is addressing major urgent needs for cancer treatment. We conducted a study to compare the frequency of 3-(2-deoxy-β-d-erythro-pentafuranosyl)pyrimido[1,2-α]purin-10(3H)-one deoxyguanosine (M1dG) and 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) adducts, biomarkers of oxidative stress and/or lipid peroxidation, on human hepatocarcinoma HepG2 cells exposed to increasing levels of Fe3O4-nanoparticles (NPs) versus untreated cells at different lengths of incubations, and in the presence of increasing exposures to an alternating magnetic field (AMF) of 186 kHz using 32P-postlabeling. The levels of oxidative damage tended to increase significantly after ≥24 h of incubations compared to controls. The oxidative DNA damage tended to reach a steady-state after treatment with 60 μg/mL of Fe3O4-NPs. Significant dose–response relationships were observed. A greater adduct production was observed after magnetic hyperthermia, with the highest amounts of oxidative lesions after 40 min exposure to AMF. The effects of magnetic hyperthermia were significantly increased with exposure and incubation times. Most important, the levels of oxidative lesions in AMF exposed NP treated cells were up to 20-fold greater relative to those observed in nonexposed NP treated cells. Generation of oxidative lesions may be a mechanism by which magnetic hyperthermia induces cancer cell death.
- Published
- 2017
41. The orphan nuclear receptor COUP-TFII coordinates hypoxia-independent proangiogenic responses in hepatic stellate cells
- Author
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Massimo Pinzani, Simone Polvani, Duccio Cavalieri, G. Marroncini, Mireille Vasseur-Cognet, S. Tempesti, Andrea Galli, Tommaso Mello, Stefano Milani, Luca Beltrame, Mirko Tarocchi, and Elisabetta Ceni
- Subjects
0301 basic medicine ,Liver Cirrhosis ,medicine.medical_specialty ,Angiogenesis ,Neovascularization, Physiologic ,Cell Communication ,Biology ,COUP Transcription Factor II ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Cell Movement ,Internal medicine ,medicine ,Hepatic Stellate Cells ,Animals ,Humans ,Hypoxia ,COUP-TFII ,Cell Proliferation ,Orphan receptor ,Mice, Knockout ,Wound Healing ,Hepatology ,Transdifferentiation ,Endothelial Cells ,NF-κB ,Transfection ,Coculture Techniques ,Cell biology ,Up-Regulation ,Mice, Inbred C57BL ,030104 developmental biology ,Endocrinology ,Nuclear receptor ,chemistry ,Liver ,HIF ,Hepatic wound healing ,NF-kB ,Notch ,angiogenesis ,fibrosis ,hepatic stellate cells ,hypoxia ,030220 oncology & carcinogenesis ,Cell Transdifferentiation ,Hepatic stellate cell ,Hepatocytes - Abstract
Background & Aims Hepatic stellate cell (HSC) transdifferentiation into collagen-producing myofibroblasts is a key event in hepatic fibrogenesis, but the transcriptional network that controls the acquisition of the activated phenotype is still poorly understood. In this study, we explored whether the nuclear receptor chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII) is involved in HSC activation and in the multifunctional role of these cells during the response to liver injury. Methods COUP-TFII expression was evaluated in normal and cirrhotic livers by immunohistochemistry and Western blot. The role of COUP-TFII in HSC was assessed by gain and loss of function transfection experiments and by generation of mice with COUP-TFII deletion in HSC. Molecular changes were determined by gene expression microarray and RT-qPCR. Results We showed that COUP-TFII is highly expressed in human fibrotic liver and in mouse models of hepatic injury. COUP-TFII expression rapidly increased upon HSC activation and it was associated with the regulation of genes involved in cell motility, proliferation and angiogenesis. Inactivation of COUP-TFII impairs proliferation and invasiveness in activated HSC and COUP-TFII deletion in mice abrogate HSC activation and angiogenesis. Finally, co-culture experiments with HSC and liver sinusoidal endothelial cells (SEC) showed that COUP-TFII expression in HSC influenced SEC migration and tubulogenesis via a hypoxia-independent and nuclear factor kappaB-dependent mechanism. Conclusion This study elucidates a novel transcriptional pathway in HSC that is involved in the acquisition of the proangiogenic phenotype and regulates the paracrine signals between HSC and SEC during hepatic wound healing. Lay summary In this study, we identified an important regulator of HSC pathobiology. We showed that the orphan receptor COUP-TFII is an important player in hepatic neoangiogenesis. COUP-TFII expression in HSC controls the crosstalk between HSC and endothelial cells coordinating vascular remodelling during liver injury. Transcript profiling ArrayExpress accession E-MTAB-1795.
- Published
- 2017
42. Rapamycin inhibits mTOR/p70S6K activation in CA3 region of the hippocampus of the rat and impairs long term memory
- Author
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Maria Grazia Giovannini, Gary L. Wenk, Daniele Lana, Tommaso Mello, and J. Di Russo
- Subjects
0301 basic medicine ,Male ,Memory, Long-Term ,Cognitive Neuroscience ,Scopolamine ,Hippocampus ,Experimental and Cognitive Psychology ,mTORC1 ,Muscarinic Antagonists ,Nicotinic Antagonists ,Mecamylamine ,mTORC2 ,Article ,03 medical and health sciences ,Behavioral Neuroscience ,0302 clinical medicine ,medicine ,Avoidance Learning ,Animals ,Phosphorylation ,Rats, Wistar ,PI3K/AKT/mTOR pathway ,Sirolimus ,Chemistry ,Long-term memory ,TOR Serine-Threonine Kinases ,Ribosomal Protein S6 Kinases, 70-kDa ,CA3 Region, Hippocampal ,Rats ,Acetylcholine ,Long term memory ,Muscarinic receptor ,Nicotinic receptor ,030104 developmental biology ,Memory, Short-Term ,Cholinergic ,Neuroscience ,030217 neurology & neurosurgery ,medicine.drug - Abstract
The present study was aimed at establishing whether the mTOR pathway and its downstream effector p70S6K in CA3 pyramidal neurons are under the modulation of the cholinergic input to trigger the formation of long term memories, similar to what we demonstrated in CA1 hippocampus. We performed in vivo behavioral experiments using the step down inhibitory avoidance test in adult Wistar rats to evaluate memory formation under different conditions. We examined the effects of rapamycin, an inhibitor of mTORC1 formation, scopolamine, a muscarinic receptor antagonist or mecamylamine, a nicotinic receptor antagonist, on short and long term memory formation and on the functionality of the mTOR pathway. Acquisition was conducted 30 min after i.c.v. injection of rapamycin. Recall testing was performed 1h, 4h or 24h after acquisition. We found that (1) mTOR and p70S6K activation in CA3 pyramidal neurons were involved in long term memory formation; (2) rapamycin significantly inhibited mTOR and of p70S6K activation at 4h, and long term memory impairment 24h after acquisition; (3) scopolamine impaired short but not long term memory, with an early increase of mTOR/p70S6K activation at 1h followed by stabilization at longer times; (4) mecamylamine and scopolamine co-administration impaired short term memory at 1h and 4h and reduced the scopolamine-induced increase of mTOR/p70S6K activation at 1h and 4h; (5) mecamylamine and scopolamine treatment did not impair long term memory formation; (6) unexpectedly, rapamycin increased mTORC2 activation in microglial cells. Our results demonstrate that in CA3 pyramidal neurons the mTOR/p70S6K pathway is under the modulation of the cholinergic system and is involved in long-term memory encoding, and are consistent with the hypothesis that the CA3 region of the hippocampus is involved in memory mechanisms based on rapid, one-trial object–place learning and recall. Furthermore, our results are in accordance with previous reports that selective molecular mechanisms underlie either short term memory, long term memory, or both. Furthermore, our discovery that administration of rapamycin increased the activation of mTORC2 in microglial cells supports a reappraisal of the beneficial/adverse effects of rapamycin administration.
- Published
- 2017
43. CHK2 overexpression and mislocalisation within mitotic structures enhances chromosomal instability and hepatocellular carcinoma progression
- Author
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Andrea Galli, Matteo Lulli, Massimo Pinzani, Andrew M. Hall, S. Madiai, Vinicio Carloni, Tommaso Mello, Tu Vinh Luong, and Krista Rombouts
- Subjects
0301 basic medicine ,Genome instability ,Pathology ,Cytoplasm ,medicine.disease_cause ,Retinoblastoma Protein ,environment and public health ,Liver Neoplasms, Experimental ,Chromosome instability ,Databases, Genetic ,Aurora Kinase B ,Diethylnitrosamine ,Phosphorylation ,Checkpoint Kinase 2 ,Liver Neoplasms ,Gastroenterology ,DNA, Neoplasm ,Up-Regulation ,GENETIC INSTABILITY ,HEPATOBILIARY CANCER ,Liver ,Premature chromosome condensation ,Gene Knockdown Techniques ,Female ,biological phenomena, cell phenomena, and immunity ,medicine.medical_specialty ,Carcinoma, Hepatocellular ,animal structures ,DNA damage ,Aurora B kinase ,Mitosis ,Spindle Apparatus ,Biology ,03 medical and health sciences ,Chromosomal Instability ,medicine ,Animals ,Humans ,Rats, Wistar ,neoplasms ,Cell Nucleus ,Biological Transport ,HCT116 Cells ,enzymes and coenzymes (carbohydrates) ,030104 developmental biology ,Cancer research ,Carcinogens ,Hepatocytes ,Carcinogenesis ,DNA Damage - Abstract
ObjectiveChromosomal instability (CIN) is the most common form of genomic instability, which promotes hepatocellular carcinoma (HCC) progression by enhancing tumour heterogeneity, drug resistance and immunity escape. CIN per se is an important factor of DNA damage, sustaining structural chromosome abnormalities but the underlying mechanisms are unknown.DesignDNA damage response protein checkpoint kinase 2 (Chk2) expression was evaluated in an animal model of diethylnitrosamine-induced HCC characterised by DNA damage and elevated mitotic errors. Chk2 was also determined in two discrete cohorts of human HCC specimens. To assess the functional role of Chk2, gain on and loss-of-function, mutagenesis, karyotyping and immunofluorescence/live imaging were performed by using HCT116, Huh7 and human hepatocytes immortalised with hTERT gene (HuS).ResultsWe demonstrate that mitotic errors during HCC tumorigenesis cause lagging chromosomes/DNA damage and activation of Chk2. Overexpression/phosphorylation and mislocalisation within the mitotic spindle of Chk2 contributes to induce lagging chromosomes. Lagging chromosomes and mitotic activity are reversed by knockdown of Chk2. Furthermore, upregulated Chk2 maintains mitotic activity interacting with Aurora B kinase for chromosome condensation and cytokinesis. The forkhead-associated domain of Chk2 is required for Chk2 mislocalisation to mitotic structures. In addition, retinoblastoma protein phosphorylation contributes to defective mitoses. A cohort and independent validation cohort show a strong cytoplasm to nuclear Chk2 translocation in a subset of patients with HCC.ConclusionsThe study reveals a new mechanistic insight in the coinvolvement of Chk2 in HCC progression. These findings propose Chk2 as a putative biomarker to detect CIN in HCC providing a valuable support for clinical/therapeutical management of patients.
- Published
- 2017
44. PARP-1 activation causes neuronal death in the hippocampal CA1 region by increasing the expression of Ca2+-permeable AMPA receptors
- Author
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Elisa Landucci, F. Moroni, Domenico E. Pellegrini-Giampietro, Francesco Resta, Elisabetta Gerace, Tommaso Mello, Guido Mannaioni, Alessio Masi, and Roberta Felici
- Subjects
Cell death ,Programmed cell death ,Poly ADP ribose polymerase ,Poly (ADP-Ribose) Polymerase-1 ,TRPM Cation Channels ,AMPA receptor ,Biology ,Poly(ADP-ribose) Polymerase Inhibitors ,Receptors, N-Methyl-D-Aspartate ,Hippocampus ,PARP ,lcsh:RC321-571 ,Tissue Culture Techniques ,chemistry.chemical_compound ,MNNG ,Animals ,TRPM2 ,Receptors, AMPA ,Rats, Wistar ,Hypoxia ,CA1 Region, Hippocampal ,lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry ,PI3K/AKT/mTOR pathway ,Pyramidal Cells ,Glutamate receptor ,Organotypic hippocampal slices ,CA3 Region, Hippocampal ,Cell biology ,Glucose ,Neurology ,Biochemistry ,chemistry ,nervous system ,Caspases ,Nerve Degeneration ,NMDA receptor ,NBQX ,Calcium ,Poly(ADP-ribose) Polymerases ,GluA1 - Abstract
An excessive activation of poly(ADP-ribose) polymerases (PARPs) may trigger a form of neuronal death similar to that occurring in neurodegenerative disorders. To investigate this process, we exposed organotypic hippocampal slices to N-methyl-N'-nitro-N'-nitrosoguanidine (MNNG, 100μM for 5min), an alkylating agent widely used to activate PARP-1. MNNG induced a pattern of degeneration of the CA1 pyramidal cells morphologically similar to that observed after a brief period of oxygen and glucose deprivation (OGD). MNNG exposure was also associated with a dramatic increase in PARP-activity and a robust decrease in NAD(+) and ATP content. These effects were prevented by PARP-1 but not PARP-2 inhibitors. In our experimental conditions, cell death was not mediated by AIF translocation (parthanatos) or caspase-dependent apoptotic processes. Furthermore, we found that PARP activation was followed by a significant deterioration of neuronal membrane properties. Using electrophysiological recordings we firstly investigated the suggested ability of ADP-ribose to open TRPM2 channels in MNNG-induced cells death, but the results we obtained showed that TRPM2 channels are not involved. We then studied the involvement of glutamate receptor-ion channel complex and we found that NBQX, a selective AMPA receptor antagonist, was able to effectively prevent CA1 neuronal loss while MK801, a NMDA antagonist, was not active. Moreover, we observed that MNNG treatment increased the ratio of GluA1/GluA2 AMPAR subunit expression, which was associated with an inward rectification of the IV relationship of AMPA sEPSCs in the CA1 but not in the CA3 subfield. Accordingly, 1-naphthyl acetyl spermine (NASPM), a selective blocker of Ca(2+)-permeable GluA2-lacking AMPA receptors, reduced MNNG-induced CA1 pyramidal cell death. In conclusion, our results show that activation of the nuclear enzyme PARP-1 may change the expression of membrane proteins and Ca(2+) permeability of AMPA channels, thus affecting the function and survival of CA1 pyramidal cells.
- Published
- 2014
45. THU-447-DNA damage response CHK2 activates senescence cellular program and supports oxidative metabolism to drive hepatocellular carcinoma development
- Author
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Vinicio Carloni, Caecilia H. C. Sukowati, Krista Rombouts, Laura Gragnani, Veronica Ghini, Andrea Galli, Matteo Lulli, Tommaso Mello, and Lorenzo Cavallini
- Subjects
Senescence ,Oxidative metabolism ,Hepatology ,DNA damage ,Hepatocellular carcinoma ,medicine ,Cancer research ,Biology ,medicine.disease - Published
- 2019
46. THU-449-Nuclear orphan receptor COUP-TF2 increases the resistence to anoikis and induces amoeboid movment and metastatic potential in hepatocellular carcinoma
- Author
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Mireille Vasseur-Cognet, Stefano Milani, Andrea Galli, Elisabetta Ceni, Mirko Tarocchi, Simone Polvani, and Tommaso Mello
- Subjects
Orphan receptor ,Hepatology ,business.industry ,Hepatocellular carcinoma ,Cancer research ,Medicine ,Anoikis ,business ,medicine.disease - Published
- 2019
47. P.04.4 NUCLEAR ORPHAN RECEPTOR COUP-TF2 INCREASED THE RESISTANCE TO ANIKOISIS AND THE METASTATIC POTENTIAL IN HEPATOCELLULAR CARCINOMA
- Author
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Elisabetta Ceni, Andrea Galli, Simone Polvani, Gabriele Dragoni, I. Simeone, Stefano Milani, L. Ciambellotti, and Tommaso Mello
- Subjects
Orphan receptor ,Hepatology ,business.industry ,Hepatocellular carcinoma ,Gastroenterology ,Cancer research ,Medicine ,business ,medicine.disease - Published
- 2019
48. Nuclear orphan receptor COUP-TF2 induces anoikis resistance, amoeboid migration and metastatic potential in hepatocellular carcinoma (HCC)
- Author
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Andrea Galli, Simone Polvani, Mireille Vasseur-Cognet, Mirko Tarocchi, Tommaso Mello, Stefano Milani, and Elisabetta Ceni
- Subjects
Orphan receptor ,Nuclear orphan receptor, COUP-TF2, anoikis resistance, amoeboid migration, metastatic potential, hepatocellular carcinoma (HCC) ,Hepatology ,business.industry ,Hepatocellular carcinoma ,Anoikis resistance ,Gastroenterology ,Cancer research ,Medicine ,business ,medicine.disease - Published
- 2019
49. A new regulator of mitochondrial OXPHOS in hepatocytes and HCC cell lines: the AAA+ ATPase RuvBL1
- Author
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Simone Polvani, Tommaso Mello, Elisabetta Ceni, I. Simeone, Andrea Galli, M. Materozzi, and Mirko Tarocchi
- Subjects
Hepatology ,Cell culture ,business.industry ,mitochondrial OXPHOS, HCC, AAA+ ATPase, RuvBL1 ,Gastroenterology ,Regulator ,Medicine ,Oxidative phosphorylation ,business ,AAA proteins ,Cell biology - Published
- 2019
50. COUP-TFII in pancreatic adenocarcinoma: Clinical implication for patient survival and tumor progression
- Author
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Andrea Galli, F. Buccoliero, Mirko Tarocchi, Marco Farsi, Gabriella Nesi, Vieri Boddi, Stefano Milani, Simone Polvani, S. Tempesti, Elisabetta Ceni, Massimo D'Amico, Silvia Nesi, and Tommaso Mello
- Subjects
Oncology ,Cancer Research ,medicine.medical_specialty ,Angiogenesis ,Biology ,medicine.disease ,Tumor progression ,Pancreatic tumor ,Internal medicine ,Pancreatic cancer ,Cancer cell ,medicine ,Adenocarcinoma ,Gene silencing ,CA19-9 - Abstract
Despite the accumulating knowledge of alterations in pancreatic cancer molecular pathways, no substantial improvements in the clinical prognosis have been made and this malignancy continues to be a leading cause of cancer death in the Western World. The orphan nuclear receptor COUP-TFII is a regulator of a wide range of biological processes and it may exert a pro-oncogenic role in cancer cells; interestingly, indirect evidences suggest that the receptor could be involved in pancreatic cancer. The aim of this study was to evaluate the expression of COUP-TFII in human pancreatic tumors and to unveil its role in the regulation of pancreatic tumor growth. We evaluated COUP-TFII expression by immunohistochemistry on primary samples. We analyzed the effect of the nuclear receptor silencing in human pancreatic cancer cells by means of shRNA expressing cell lines. We finally confirmed the in vitro results by in vivo experiments on nude mice. COUP-TFII is expressed in 69% of tested primary samples and correlates with the N1 and M1 status and clinical stage; Kaplan-Meier and Cox regression analysis show that it may be an independent prognostic factor of worst outcome. In vitro silencing of COUP-TFII reduces the cell growth and invasiveness and it strongly inhibits angiogenesis, an effect mediated by the regulation of VEGF-C. In nude mice, COUP-TFII silencing reduces tumor growth by 40%. Our results suggest that COUP-TFII might be an important regulator of the behavior of pancreatic adenocarcinoma, thus representing a possible new target for pancreatic cancer therapy.
- Published
- 2013
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