46 results on '"Serena Vella"'
Search Results
2. Cisplatin Resistance in Osteosarcoma: In vitro Validation of Candidate DNA Repair-Related Therapeutic Targets and Drugs for Tailored Treatments
- Author
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Marilù Fanelli, Elisa Tavanti, Maria Pia Patrizio, Serena Vella, Amira Fernandez-Ramos, Federica Magagnoli, Silvia Luppi, Claudia Maria Hattinger, and Massimo Serra
- Subjects
osteosarcoma ,DNA repair ,cisplatin ,drug resistance ,chemotherapy ,targeted drugs ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Treatment of high-grade osteosarcoma, the most common malignant tumor of bone, is largely based on administration of cisplatin and other DNA damaging drugs. Altered DNA repair mechanisms may thus significantly impact on either response or resistance to chemotherapy. In this study, by using a panel of human osteosarcoma cell lines, either sensitive or resistant to cisplatin, we assessed the value as candidate therapeutic targets of DNA repair-related factors belonging to the nucleotide excision repair (NER) or base excision repair (BER) pathways, as well as of a group of 18 kinases, which expression was higher in cisplatin-resistant variants compared to their parental cell lines and may be indirectly involved in DNA repair. The causal involvement of these factors in cisplatin resistance of human osteosarcoma cells was validated through gene silencing approaches and in vitro reversal of CDDP resistance. This approach highlighted a subgroup of genes, which value as promising candidate therapeutic targets was further confirmed by protein expression analyses. The in vitro activity of 15 inhibitor drugs against either these genes or their pathways was then analyzed, in order to identify the most active ones in terms of inherent activity and ability to overcome cisplatin resistance. NSC130813 (NERI02; F06) and triptolide, both targeting NER factors, proved to be the two most active agents, without evidence of cross-resistance with cisplatin. Combined in vitro treatments showed that NSC130813 and triptolide, when administered together with cisplatin, were able to improve its efficacy in both drug-sensitive and resistant osteosarcoma cells. This evidence may indicate an interesting therapeutic future option for treatment of osteosarcoma patients who present reduced responsiveness to cisplatin, even if possible effects of additive collateral toxicities must be carefully considered. Moreover, our study also showed that targeting protein kinases belonging to the mitogen-activated protein kinase (MAPK) or fibroblast growth factor receptor (FGFR) pathways might indicate new promising therapeutic perspectives in osteosarcoma, demanding for additional investigation.
- Published
- 2020
- Full Text
- View/download PDF
3. Small Extracellular Vesicles from Human Fetal Dermal Cells and Their MicroRNA Cargo: KEGG Signaling Pathways Associated with Angiogenesis and Wound Healing
- Author
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Cinzia Maria Chinnici, Giandomenico Amico, Alessia Gallo, Gioacchin Iannolo, Nicola Cuscino, Serena Vella, Claudia Carcione, David Nascari, and Pier Giulio Conaldi
- Subjects
Internal medicine ,RC31-1245 - Abstract
The use of cell secreted factors in clinical settings could be an alternative to conventional cell therapy, with the advantage of limiting concerns generally associated with traditional cell transplantation, such as tumorigenicity, immunoreactivity, and carrying of infections. Based on our published data, we predict a potential role for extracellular vesicles (EVs) in contributing to the proangiogenic activity of human fetal dermal cell secretome. Depletion of nanosized EVs from secretome significantly impaired its ability to induce formation of mesh-like structures in vitro. The isolated EVs were characterized for size and concentration by nanoparticle tracking analysis, and for protein markers (Rab5+, Alix+, CD63+, and calnexin-). The microRNA profile of EVs revealed 87 microRNAs significantly upregulated (≥15-fold increase) in fetal compared to adult dermal cell-derived EVs. Interestingly, these upregulated microRNAs included microRNAs with a validated role in angiogenesis according to literature. Moreover, the DIANA-TarBase v7.0 analysis confirmed enrichment in the KEGG signaling pathways associated with angiogenesis and wound healing, with the identification of putative target genes including thrombospondin 1. To validate the in silico data, EVs were also characterized for total protein contents. When tested in in vitro angiogenesis, fetal dermal cell-derived EVs were more effective than their adult counterpart in inducing formation of complete mesh-like structures. Furthermore, treatment of fibroblasts with fetal dermal-derived EVs determined a 4-fold increase of thrombospondin 1 protein amounts compared with the untreated fibroblasts. Finally, visualization of CSFE-labeled EVs in the cytosol of target cells suggested a successful uptake of these particles at 4-8 hours of incubation. We conclude that EVs are important contributors of the proangiogenic effect of fetal dermal cell secretome. Hence, EVs could also serve as vehicle for a successful delivery of microRNAs or other molecules of therapeutic interest to target cells.
- Published
- 2020
- Full Text
- View/download PDF
4. Lung resident mesenchymal cells isolated from patients with the Bronchiolitis Obliterans Syndrome display a deregulated epigenetic profile
- Author
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Serena Vella, Pier Giulio Conaldi, Emanuela Cova, Federica Meloni, Rosa Liotta, Salvatore Cuzzocrea, Lavinia Martino, Alessandro Bertani, Angelo Luca, and Patrizio Vitulo
- Subjects
Medicine ,Science - Abstract
Abstract Bronchiolitis Obliterans Syndrome is the major determinant of the graft function loss after lung transplantation, but its pathogenesis is still incompletely understood and currently available therapeutic strategies are poorly effective. A deeper understanding of its pathogenic mechanisms is crucial for the development of new strategies to prevent and treat this devastating complication. In this study, we focused on the mesenchymal stromal cells, recently recognized as BOS key effectors, and our primary aim was to identify their epigenetic determinants, such as histone modifications and non-coding RNA regulation, which could contribute to their differentiation in myofibroblasts. Interestingly, we identified a deregulated expression of histone deacetylases and methyltransferases, and a microRNA-epigenetic regulatory network, which could represent novel targets for anti-fibrotic therapy. We validated our results in vitro, in a cell model of fibrogenesis, confirming the epigenetic involvement in this process and paving the way for a new application for epigenetic drugs.
- Published
- 2018
- Full Text
- View/download PDF
5. Comparison of Immunosuppressive and Angiogenic Properties of Human Amnion-Derived Mesenchymal Stem Cells between 2D and 3D Culture Systems
- Author
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Vitale Miceli, Mariangela Pampalone, Serena Vella, Anna Paola Carreca, Giandomenico Amico, and Pier Giulio Conaldi
- Subjects
Internal medicine ,RC31-1245 - Abstract
The secretion of potential therapeutic factors by mesenchymal stem cells (MSCs) has aroused much interest given the benefits that it can bring in the field of regenerative medicine. Indeed, the in vitro multipotency of these cells and the secretive capacity of both angiogenic and immunomodulatory factors suggest a role in tissue repair and regeneration. However, during culture, MSCs rapidly lose the expression of key transcription factors associated with multipotency and self-renewal, as well as the ability to produce functional paracrine factors. In our study, we show that a three-dimensional (3D) culture method is effective to induce MSC spheroid formation, to maintain the multipotency and to improve the paracrine activity of a specific population of human amnion-derived MSCs (hAMSCs). The regenerative potential of both 3D culture-derived conditioned medium (3D CM) and their exosomes (EXO) was assessed against 2D culture products. In particular, tubulogenesis assays revealed increased capillary maturation in the presence of 3D CM compared with both 2D CM and 2D EXO. Furthermore, 3D CM had a greater effect on inhibition of PBMC proliferation than both 2D CM and 2D EXO. To support this data, hAMSC spheroids kept in our 3D culture system remained viable and multipotent and secreted considerable amounts of both angiogenic and immunosuppressive factors, which were detected at lower levels in 2D cultures. This work reveals the placenta as an important source of MSCs that can be used for eventual clinical applications as cell-free therapies.
- Published
- 2019
- Full Text
- View/download PDF
6. A One-Pot Approach to Novel Pyridazine C-Nucleosides
- Author
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Flavio Cermola, Serena Vella, Marina DellaGreca, Angela Tuzi, and Maria Rosaria Iesce
- Subjects
glycosyl furans ,singlet oxygen ,[4+2] cycloaddition ,C-nucleosides ,photooxygenation ,pyridazines ,Organic chemistry ,QD241-441 - Abstract
The synthesis of glycosides and modified nucleosides represents a wide research field in organic chemistry. The classical methodology is based on coupling reactions between a glycosyl donor and an acceptor. An alternative strategy for new C-nucleosides is used in this approach, which consists of modifying a pre-existent furyl aglycone. This approach is applied to obtain novel pyridazine C-nucleosides starting with 2- and 3-(ribofuranosyl)furans. It is based on singlet oxygen [4+2] cycloaddition followed by reduction and hydrazine cyclization under neutral conditions. The mild three-step one-pot procedure leads stereoselectively to novel pyridazine C-nucleosides of pharmacological interest. The use of acetyls as protecting groups provides an elegant direct route to a deprotected new pyridazine C-nucleoside.
- Published
- 2021
- Full Text
- View/download PDF
7. Co-Administration of Fendiline Hydrochloride Enhances Chemotherapeutic Efficacy of Cisplatin in Neuroblastoma Treatment
- Author
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Antonella Brizzolara, Patrizia Garbati, Serena Vella, Matilde Calderoni, Alessandro Quattrone, Gian Paolo Tonini, Mario Capasso, Luca Longo, Raffaella Barbieri, Tullio Florio, and Aldo Pagano
- Subjects
neuroblastoma ,non-coding RNA ,NDM29 ,fendiline hydrochloride ,GD2 ,Organic chemistry ,QD241-441 - Abstract
Despite significant improvement of neuroblastoma (NB) patients’ survival due to recent treatment advancements in recent years, NB is still associated with high mortality rate. In search of novel strategies to increase NB’s susceptibility to pharmacological treatments, we investigated the in vitro and in vivo effects of fendiline hydrochloride as an enhancer of cisplatin antitumor activity. To assess the modulation of fendiline treatment on cisplatin responses, we used in vitro (evaluating NB cell proliferation by XCELLigence technology and colony formation, and gene expression by RT-PCR) and in vivo (NB cell grafts in NOD-SCID mice) models of NB. NB cell treatment with fendiline induced the expression of the ncRNA NDM29, leading to cell differentiation and to the reduction of the expression of MDRs/ABC transporters linked to multidrug resistance. These events were correlated to higher NB cell susceptibility to cisplatin and, consequently, increased its cytotoxic potency. In vivo, this drug interaction causes an enhanced ability of cisplatin to induce apoptosis in NB masses, resulting in tumor growth reduction and prolonged animal survival rate. Thus, the administration of fendiline might be a possible novel therapeutic approach to increase cisplatin efficacy in aggressive and poorly responsive NB cases.
- Published
- 2020
- Full Text
- View/download PDF
8. PPAR Gamma in Neuroblastoma: The Translational Perspectives of Hypoglycemic Drugs
- Author
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Serena Vella, Pier Giulio Conaldi, Tullio Florio, and Aldo Pagano
- Subjects
Biology (General) ,QH301-705.5 - Abstract
Neuroblastoma (NB) is the most common and aggressive pediatric cancer, characterized by a remarkable phenotypic diversity and high malignancy. The heterogeneous clinical behavior, ranging from spontaneous remission to fatal metastatic disease, is attributable to NB biology and genetics. Despite major advances in therapies, NB is still associated with a high morbidity and mortality. Thus, novel diagnostic, prognostic, and therapeutic approaches are required, mainly to improve treatment outcomes of high-risk NB patients. Among neuroepithelial cancers, NB is the most studied tumor as far as PPAR ligands are concerned. PPAR ligands are endowed with antitumoral effects, mainly acting on cancer stem cells, and constitute a possible add-on therapy to antiblastic drugs, in particular for NB with unfavourable prognosis. While discussing clinical background, this review will provide a synopsis of the major studies about PPAR expression in NB, focusing on the potential beneficial effects of hypoglycemic drugs, thiazolidinediones and metformin, to reduce the occurrence of relapses as well as tumor regrowth in NB patients.
- Published
- 2016
- Full Text
- View/download PDF
9. Targeting CDKs with Roscovitine Increases Sensitivity to DNA Damaging Drugs of Human Osteosarcoma Cells.
- Author
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Serena Vella, Elisa Tavanti, Claudia Maria Hattinger, Marilù Fanelli, Rogier Versteeg, Jan Koster, Piero Picci, and Massimo Serra
- Subjects
Medicine ,Science - Abstract
Cyclin-dependent kinase 2 (CDK2) has been reported to be essential for cell proliferation in several human tumours and it has been suggested as an appropriate target to be considered in order to enhance the efficacy of treatment regimens based on the use of DNA damaging drugs. We evaluated the clinical impact of CDK2 overexpression on a series of 21 high-grade osteosarcoma (OS) samples profiled by using cDNA microarrays. We also assessed the in vitro efficacy of the CDKs inhibitor roscovitine in a panel of drug-sensitive and drug-resistant human OS cell lines. OS tumour samples showed an inherent overexpression of CDK2, and high expression levels at diagnosis of this kinase appeared to negatively impact on clinical outcome. CDK2 expression also proved to be relevant for in vitro OS cells growth. These findings indicated CDK2 as a promising candidate therapeutic marker for OS and therefore we assessed the efficacy of the CDKs-inhibitor roscovitine in both drug-sensitive and -resistant OS cell lines. All cell lines resulted to be responsive to roscovitine, which was also able to increase the activity of cisplatin and doxorubicin, the two most active DNA damaging drugs used in OS chemotherapy. Our results indicated that combined treatment with conventional OS chemotherapeutic drugs and roscovitine may represent a new candidate intervention approach, which may be considered to enhance tumour cell sensitivity to DNA damaging drugs.
- Published
- 2016
- Full Text
- View/download PDF
10. Hyper-activation of Notch3 amplifies the proliferative potential of rhabdomyosarcoma cells.
- Author
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Maria De Salvo, Lavinia Raimondi, Serena Vella, Laura Adesso, Roberta Ciarapica, Federica Verginelli, Antonio Pannuti, Arianna Citti, Renata Boldrini, Giuseppe M Milano, Antonella Cacchione, Andrea Ferrari, Paola Collini, Angelo Rosolen, Gianni Bisogno, Rita Alaggio, Alessandro Inserra, Mattia Locatelli, Stefano Stifani, Isabella Screpanti, Lucio Miele, Franco Locatelli, and Rossella Rota
- Subjects
Medicine ,Science - Abstract
Rhabdomyosarcoma (RMS) is a pediatric myogenic-derived soft tissue sarcoma that includes two major histopathological subtypes: embryonal and alveolar. The majority of alveolar RMS expresses PAX3-FOXO1 fusion oncoprotein, associated with the worst prognosis. RMS cells show myogenic markers expression but are unable to terminally differentiate. The Notch signaling pathway is a master player during myogenesis, with Notch1 activation sustaining myoblast expansion and Notch3 activation inhibiting myoblast fusion and differentiation. Accordingly, Notch1 signaling is up-regulated and activated in embryonal RMS samples and supports the proliferation of tumor cells. However, it is unable to control their differentiation properties. We previously reported that Notch3 is activated in RMS cell lines, of both alveolar and embryonal subtype, and acts by inhibiting differentiation. Moreover, Notch3 depletion reduces PAX3-FOXO1 alveolar RMS tumor growth in vivo. However, whether Notch3 activation also sustains the proliferation of RMS cells remained unclear. To address this question, we forced the expression of the activated form of Notch3, Notch3IC, in the RH30 and RH41 PAX3-FOXO1-positive alveolar and in the RD embryonal RMS cell lines and studied the proliferation of these cells. We show that, in all three cell lines tested, Notch3IC over-expression stimulates in vitro cell proliferation and prevents the effects of pharmacological Notch inhibition. Furthermore, Notch3IC further increases RH30 cell growth in vivo. Interestingly, knockdown of Notch canonical ligands JAG1 or DLL1 in RMS cell lines decreases Notch3 activity and reduces cell proliferation. Finally, the expression of Notch3IC and its target gene HES1 correlates with that of the proliferative marker Ki67 in a small cohort of primary PAX-FOXO1 alveolar RMS samples. These results strongly suggest that high levels of Notch3 activation increase the proliferative potential of RMS cells.
- Published
- 2014
- Full Text
- View/download PDF
11. NMR as powerful technology for noninvasively monitoring cell health and expansion during bioprocessing
- Author
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Francesca Benevelli, Serena Vella, Chiara Crosta, Elena Demetrio, Christian Fischer, Marco Pupo, and Stefano Baila
- Subjects
Bioreactors ,Magnetic Resonance Spectroscopy ,Cell Culture Techniques ,Cell- and Tissue-Based Therapy ,Bioengineering ,Applied Microbiology and Biotechnology ,Cell Proliferation ,Biotechnology - Abstract
Over the last decades, the success of advanced cell therapies and the increasing production volumes of vaccines, proteins, or viral vectors have raised the need of robust cell-based manufacturing processes for ensuring product quality and satisfying good manufacturing practice requirements. The cultivation process of cells needs to be highly controlled for improved productivity, reduced variability, and optimized bioprocesses. Cell cultures can be easily monitored using different technologies, which could deliver direct or indirect assessment of the cells' viability. Among these techniques, nuclear magnetic resonance (NMR) spectroscopy is a powerful technology that permits the evaluation and the identification of key endogenous metabolites. NMR can provide information on the cell metabolic pathways, on the bioprocesses, and is also capable to quickly test for impurities. In this study, NMR was successfully used as a technology for monitoring cell viability and expansion in different supports for cell growth (including bioreactors), to predict the bioprocess output and for the early identification of key metabolites linked to cell starvation. This investigation will allow the timely control of culture conditions and favor the optimization of the bioprocesses.
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- 2022
- Full Text
- View/download PDF
12. Book of abstracts Chimici per le Biotecnologie 2023
- Author
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Ciarleglio, Gianluca, Domenico, Montesano, Serena, Vella, Gilda, Benevento, Toto, Elisa, and Santonicola, Mariagabriella
- Subjects
microspheres ,drug delivery systems ,stimuli-sensitive polymers ,electrospray ,hydrogels - Published
- 2023
13. Emulsion-based multi-responsive microspheres for the delivery of lipophilic Ozoile
- Author
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Gianluca Ciarleglio, Serena Vella, Elisa Toto, and M. Gabriella Santonicola
- Subjects
applications ,chemical ,Process Chemistry and Technology ,biomedical ,thermal ,Surfaces, Coatings and Films ,Electronic, Optical and Magnetic Materials ,microspheres ,extrusion ,properties ,hydrogel ,Materials Chemistry ,Ceramics and Composites - Published
- 2022
- Full Text
- View/download PDF
14. Multi-responsive hydrogel microspheres for the controlled release of ozoile
- Author
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Ciarleglio, Gianluca, Serena, Vella, Toto, Elisa, and Santonicola, Mariagabriella
- Subjects
hydrogel microspheres ,drug delivery ,temperature-responsive ,pH-responsive - Published
- 2022
15. Stimuli responsive emulsion-based micro/nanospheres for the delivery of lipophilic therapeutics
- Author
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GIANLUCA CIARLEGLIO, Serena, Vella, ELISA TOTO, and Mariagabriella Santonicola
- Subjects
hydrogel microspheres ,drug delivery ,temperature-responsive ,pH-responsive - Published
- 2022
16. Ozoile-alginate microspheres for the therapeutic management of Crohn's disease
- Author
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Ciarleglio, Gianluca, Serena, Vella, Toto, Elisa, and Santonicola, Mariagabriella
- Subjects
drug delivery systems ,hydrogel microspheres ,stimuli-responsive microspheres ,ozonides ,ozoile - Published
- 2022
17. A one-pot approach to novel pyridazine c-nucleosides
- Author
-
Angela Tuzi, Marina DellaGreca, Serena Vella, Flavio Cermola, Maria Rosaria Iesce, Cermola, Flavio, Vella, Serena, DELLA GRECA, Marina, Tuzi, Angela, and Iesce, MARIA ROSARIA
- Subjects
Hydrazine ,[4+2] cycloaddition ,Glycosyl furan ,Chemistry, Organic ,Pharmaceutical Science ,010402 general chemistry ,01 natural sciences ,Coupling reaction ,Analytical Chemistry ,Pyridazine ,chemistry.chemical_compound ,QD241-441 ,C-nucleosides ,Drug Discovery ,Furan ,Nucleoside ,Glycosides ,Physical and Theoretical Chemistry ,Furans ,Glycosyl donor ,Reduction ,010405 organic chemistry ,Singlet oxygen ,Terpenes ,Communication ,Organic Chemistry ,Nucleosides ,pyridazines ,Combinatorial chemistry ,Cycloaddition ,0104 chemical sciences ,Glycoside ,glycosyl furans ,Aglycone ,Photooxygenation ,chemistry ,Chemistry (miscellaneous) ,Molecular Medicine ,C-nucleoside - Abstract
The synthesis of glycosides and modified nucleosides represents a wide research field in organic chemistry. The classical methodology is based on coupling reactions between a glycosyl donor and an acceptor. An alternative strategy for new C-nucleosides is used in this approach, which consists of modifying a pre-existent furyl aglycone. This approach is applied to obtain novel pyridazine C-nucleosides starting with 2- and 3-(ribofuranosyl)furans. It is based on singlet oxygen [4+2] cycloaddition followed by reduction and hydrazine cyclization under neutral conditions. The mild three-step one-pot procedure leads stereoselectively to novel pyridazine C-nucleosides of pharmacological interest. The use of acetyls as protecting groups provides an elegant direct route to a deprotected new pyridazine C-nucleoside.
- Published
- 2021
18. Small Extracellular Vesicles from Human Fetal Dermal Cells and Their MicroRNA Cargo: KEGG Signaling Pathways Associated with Angiogenesis and Wound Healing
- Author
-
Giandomenico Amico, Gioacchin Iannolo, Claudia Carcione, Serena Vella, Nicola Cuscino, David Nascari, Cinzia Maria Chinnici, Pier Giulio Conaldi, and Alessia Gallo
- Subjects
Article Subject ,Angiogenesis ,Chemistry ,Cell ,Cell Biology ,RC31-1245 ,Cell biology ,Cell therapy ,medicine.anatomical_structure ,Downregulation and upregulation ,Thrombospondin 1 ,medicine ,Signal transduction ,KEGG ,Wound healing ,Internal medicine ,Molecular Biology ,Research Article - Abstract
The use of cell secreted factors in clinical settings could be an alternative to conventional cell therapy, with the advantage of limiting concerns generally associated with traditional cell transplantation, such as tumorigenicity, immunoreactivity, and carrying of infections. Based on our published data, we predict a potential role for extracellular vesicles (EVs) in contributing to the proangiogenic activity of human fetal dermal cell secretome. Depletion of nanosized EVs from secretome significantly impaired its ability to induce formation of mesh-like structures in vitro. The isolated EVs were characterized for size and concentration by nanoparticle tracking analysis, and for protein markers (Rab5+, Alix+, CD63+, and calnexin-). The microRNA profile of EVs revealed 87 microRNAs significantly upregulated (≥15-fold increase) in fetal compared to adult dermal cell-derived EVs. Interestingly, these upregulated microRNAs included microRNAs with a validated role in angiogenesis according to literature. Moreover, the DIANA-TarBase v7.0 analysis confirmed enrichment in the KEGG signaling pathways associated with angiogenesis and wound healing, with the identification of putative target genes including thrombospondin 1. To validate the in silico data, EVs were also characterized for total protein contents. When tested in in vitro angiogenesis, fetal dermal cell-derived EVs were more effective than their adult counterpart in inducing formation of complete mesh-like structures. Furthermore, treatment of fibroblasts with fetal dermal-derived EVs determined a 4-fold increase of thrombospondin 1 protein amounts compared with the untreated fibroblasts. Finally, visualization of CSFE-labeled EVs in the cytosol of target cells suggested a successful uptake of these particles at 4-8 hours of incubation. We conclude that EVs are important contributors of the proangiogenic effect of fetal dermal cell secretome. Hence, EVs could also serve as vehicle for a successful delivery of microRNAs or other molecules of therapeutic interest to target cells.
- Published
- 2020
19. Cisplatin Resistance in Osteosarcoma: In vitro Validation of Candidate DNA Repair-Related Therapeutic Targets and Drugs for Tailored Treatments
- Author
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Claudia Maria Hattinger, Silvia Luppi, Massimo Serra, Amira Fernandez-Ramos, Federica Magagnoli, Elisa Tavanti, Serena Vella, Maria Pia Patrizio, and Marilù Fanelli
- Subjects
0301 basic medicine ,MAPK/ERK pathway ,Cancer Research ,DNA repair ,targeted drugs ,cisplatin ,chemotherapy ,lcsh:RC254-282 ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,osteosarcoma ,Medicine ,Gene silencing ,tailored treatment ,Original Research ,Cisplatin ,drug resistance ,business.industry ,Base excision repair ,Triptolide ,medicine.disease ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,030104 developmental biology ,Oncology ,chemistry ,030220 oncology & carcinogenesis ,Cancer research ,Osteosarcoma ,business ,Nucleotide excision repair ,medicine.drug - Abstract
Treatment of high-grade osteosarcoma, the most common malignant tumor of bone, is largely based on administration of cisplatin and other DNA damaging drugs. Altered DNA repair mechanisms may thus significantly impact on either response or resistance to chemotherapy. In this study, by using a panel of human osteosarcoma cell lines, either sensitive or resistant to cisplatin, we assessed the value as candidate therapeutic targets of DNA repair-related factors belonging to the nucleotide excision repair (NER) or base excision repair (BER) pathways, as well as of a group of 18 kinases, which expression was higher in cisplatin-resistant variants compared to their parental cell lines and may be indirectly involved in DNA repair. The causal involvement of these factors in cisplatin resistance of human osteosarcoma cells was validated through gene silencing approaches and in vitro reversal of CDDP resistance. This approach highlighted a subgroup of genes, which value as promising candidate therapeutic targets was further confirmed by protein expression analyses. The in vitro activity of 15 inhibitor drugs against either these genes or their pathways was then analyzed, in order to identify the most active ones in terms of inherent activity and ability to overcome cisplatin resistance. NSC130813 (NERI02; F06) and triptolide, both targeting NER factors, proved to be the two most active agents, without evidence of cross-resistance with cisplatin. Combined in vitro treatments showed that NSC130813 and triptolide, when administered together with cisplatin, were able to improve its efficacy in both drug-sensitive and resistant osteosarcoma cells. This evidence may indicate an interesting therapeutic future option for treatment of osteosarcoma patients who present reduced responsiveness to cisplatin, even if possible effects of additive collateral toxicities must be carefully considered. Moreover, our study also showed that targeting protein kinases belonging to the mitogen-activated protein kinase (MAPK) or fibroblast growth factor receptor (FGFR) pathways might indicate new promising therapeutic perspectives in osteosarcoma, demanding for additional investigation.
- Published
- 2020
- Full Text
- View/download PDF
20. Co-administration of fendiline hydrochloride enhances chemotherapeutic efficacy of cisplatin in neuroblastoma treatment
- Author
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Mario Capasso, Antonella Brizzolara, Luca Longo, Serena Vella, Raffaella Barbieri, Patrizia Garbati, Tullio Florio, Alessandro Quattrone, Aldo Pagano, Matilde Calderoni, Gian Paolo Tonini, Brizzolara, A., Garbati, P., Vella, S., Calderoni, M., Quattrone, A., Tonini, G. P., Capasso, M., Longo, L., Barbieri, R., Florio, T., and Pagano, A.
- Subjects
RNA, Untranslated ,Cellular differentiation ,Cell ,Pharmaceutical Science ,Apoptosis ,Mice, SCID ,Analytical Chemistry ,Antineoplastic Agent ,Neuroblastoma ,Mice ,0302 clinical medicine ,Mice, Inbred NOD ,Antineoplastic Combined Chemotherapy Protocols ,Drug Discovery ,0303 health sciences ,Fendiline ,Brain Neoplasms ,Chemistry ,Drug Synergism ,NDM29 ,Gene Expression Regulation, Neoplastic ,GD2 ,fendiline hydrochloride ,neuroblastoma ,non-coding RNA ,medicine.anatomical_structure ,Chemistry (miscellaneous) ,030220 oncology & carcinogenesis ,Molecular Medicine ,medicine.drug ,Human ,Cell Survival ,Antineoplastic Agents ,Article ,lcsh:QD241-441 ,Brain Neoplasm ,03 medical and health sciences ,lcsh:Organic chemistry ,In vivo ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Physical and Theoretical Chemistry ,Non-coding RNA ,neoplasms ,030304 developmental biology ,Cell Proliferation ,Cisplatin ,Antineoplastic Combined Chemotherapy Protocol ,Cell growth ,Animal ,Organic Chemistry ,Apoptosi ,medicine.disease ,Fendiline hydrochloride ,Cancer research ,Neoplasm Transplantation - Abstract
Despite significant improvement of neuroblastoma (NB) patients&rsquo, survival due to recent treatment advancements in recent years, NB is still associated with high mortality rate. In search of novel strategies to increase NB&rsquo, s susceptibility to pharmacological treatments, we investigated the in vitro and in vivo effects of fendiline hydrochloride as an enhancer of cisplatin antitumor activity. To assess the modulation of fendiline treatment on cisplatin responses, we used in vitro (evaluating NB cell proliferation by XCELLigence technology and colony formation, and gene expression by RT-PCR) and in vivo (NB cell grafts in NOD-SCID mice) models of NB. NB cell treatment with fendiline induced the expression of the ncRNA NDM29, leading to cell differentiation and to the reduction of the expression of MDRs/ABC transporters linked to multidrug resistance. These events were correlated to higher NB cell susceptibility to cisplatin and, consequently, increased its cytotoxic potency. In vivo, this drug interaction causes an enhanced ability of cisplatin to induce apoptosis in NB masses, resulting in tumor growth reduction and prolonged animal survival rate. Thus, the administration of fendiline might be a possible novel therapeutic approach to increase cisplatin efficacy in aggressive and poorly responsive NB cases.
- Published
- 2020
21. A Planned and Preventive Conservation Project for The Tarot Garden by Niki de Saint Phalle
- Author
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M. Carolina Gaetani, Serena Vella, and Ulderico Santamaria
- Subjects
Sculpture ,060102 archaeology ,media_common.quotation_subject ,010401 analytical chemistry ,Art history ,0601 history and archaeology ,SAINT ,06 humanities and the arts ,Conservation ,Art ,01 natural sciences ,0104 chemical sciences ,media_common - Abstract
The Tarot Garden (Giardino dei Tarocchi), also known as The Monsters (I Mostri), is a well-known sculpture park realized between 1979 and 1998 by the French-American artist Niki de Saint Phalle (Fi...
- Published
- 2018
- Full Text
- View/download PDF
22. Comparison of Immunosuppressive and Angiogenic Properties of Human Amnion-Derived Mesenchymal Stem Cells between 2D and 3D Culture Systems
- Author
-
Pier Giulio Conaldi, Serena Vella, Mariangela Pampalone, Giandomenico Amico, Anna Paola Carreca, Vitale Miceli, Miceli V., Pampalone M., Vella S., Carreca A.P., Amico G., and Conaldi P.G.
- Subjects
lcsh:Internal medicine ,Amnion ,Article Subject ,Regeneration (biology) ,Mesenchymal stem cell ,Cell Biology ,Biology ,Regenerative medicine ,Microvesicles ,In vitro ,Cell biology ,Paracrine signalling ,medicine.anatomical_structure ,human amnion-derived mesenchymal stem cell ,medicine ,lcsh:RC31-1245 ,Molecular Biology ,Transcription factor ,Research Article - Abstract
The secretion of potential therapeutic factors by mesenchymal stem cells (MSCs) has aroused much interest given the benefits that it can bring in the field of regenerative medicine. Indeed, the in vitro multipotency of these cells and the secretive capacity of both angiogenic and immunomodulatory factors suggest a role in tissue repair and regeneration. However, during culture, MSCs rapidly lose the expression of key transcription factors associated with multipotency and self-renewal, as well as the ability to produce functional paracrine factors. In our study, we show that a three-dimensional (3D) culture method is effective to induce MSC spheroid formation, to maintain the multipotency and to improve the paracrine activity of a specific population of human amnion-derived MSCs (hAMSCs). The regenerative potential of both 3D culture-derived conditioned medium (3D CM) and their exosomes (EXO) was assessed against 2D culture products. In particular, tubulogenesis assays revealed increased capillary maturation in the presence of 3D CM compared with both 2D CM and 2D EXO. Furthermore, 3D CM had a greater effect on inhibition of PBMC proliferation than both 2D CM and 2D EXO. To support this data, hAMSC spheroids kept in our 3D culture system remained viable and multipotent and secreted considerable amounts of both angiogenic and immunosuppressive factors, which were detected at lower levels in 2D cultures. This work reveals the placenta as an important source of MSCs that can be used for eventual clinical applications as cell-free therapies.
- Published
- 2019
23. miRNA-mediated regulation of Mucin-type O-Glycosylation pathway: a putative mechanism of salivary gland dysfunction in Sjögren's syndrome
- Author
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Marta Mosca, Pier Giulio Conaldi, Chiara Baldini, Nicola Cuscino, Stefano Bombardieri, Antonella Cecchettini, Ilias Alevizos, Serena Vella, Francesco Ferro, Fabio Tuzzolino, and Alessia Gallo
- Subjects
0301 basic medicine ,Adult ,Glycosylation ,In silico ,Immunology ,Sjögren’s syndrome, MiRNA, pathogenesis, epigenetics ,Real-Time Polymerase Chain Reaction ,Salivary Glands, Minor ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Rheumatology ,Downregulation and upregulation ,microRNA ,medicine ,Immunology and Allergy ,Humans ,Computer Simulation ,Epigenetics ,Gene ,Aged ,030203 arthritis & rheumatology ,Salivary gland ,epigenetics ,business.industry ,pathogenesis ,Mucin ,Mucins ,Middle Aged ,Molecular biology ,Up-Regulation ,MicroRNAs ,030104 developmental biology ,medicine.anatomical_structure ,Sjogren's Syndrome ,chemistry ,Sjögren’s syndrome ,Female ,business ,Transcriptome ,MiRNA ,Signal Transduction - Abstract
Objective.To investigate microRNA (miRNA) that is potentially implicated in primary Sjögren syndrome (pSS)–related salivary hypofunction in labial salivary glands and to study miRNA-mediated mechanisms underlying oral dryness and altered rheology, focusing on the mucin O-glycosylation pathway.Methods.We performed miRNA expression profiling in minor salivary gland samples of patients with pSS presenting a different impairment in their unstimulated salivary flow rate. A computational in silico analysis was performed to identify genes and pathways that might be modulated by the deregulated miRNA that we had identified. To confirm in silico analysis, expression levels of genes encoding for glycosyltransferases and glycan-processing enzymes were investigated using Human Glycosylation-RT2 Profiler PCR Array.Results.Among 754 miRNA analyzed, we identified 126 miRNA that were significantly deregulated in pSS compared to controls, with a trend that was inversely proportional with the impairment of salivary flow rates. An in silico approach pinpointed that several upregulated miRNA in patients with pSS target important genes in the mucin O-glycosylation. We confirmed this prediction by quantitative real-time PCR, highlighting the downregulation of some glycosyltransferase and glycosidase genes in pSS samples compared to controls, such as GALNT1, responsible for mucin-7 glycosylation.Conclusion.Collectively, our data suggest that the expression of different predicted miRNA-target genes in the mucin type O-glycan biosynthesis pathway is altered in pSS patients with low salivary flow and that the miRNA expression profile could influence the glycosidase expression levels and consequently the rheology in pSS.
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- 2019
24. Candidate germline polymorphisms of genes belonging to the pathways of four drugs used in osteosarcoma standard chemotherapy associated with risk, survival and toxicity in non-metastatic high-grade osteosarcoma
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Sara Gagno, Erika Iacoboni, Serena Vella, Rossana Roncato, Piero Picci, Massimo Serra, Giuseppe Toffoli, Stefano Ferrari, Marilù Fanelli, Andrea Roli, Claudia Maria Hattinger, Luciana Giodini, Elisa Tavanti, Paola Biason, Katia Scotlandi, Hattinger, Claudia M., Biason, Paola, Iacoboni, Erika, Gagno, Sara, Fanelli, Marilù, Tavanti, Elisa, Vella, Serena, Ferrari, Stefano, Roli, Andrea, Roncato, Rossana, Giodini, Luciana, Scotlandi, Katia, Picci, Piero, Toffoli, Giuseppe, and Serra, Massimo
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Male ,0301 basic medicine ,Oncology ,medicine.medical_treatment ,Pharmacology ,drug response biomarkers ,Cohort Studies ,0302 clinical medicine ,XRCC3 ,Antineoplastic Combined Chemotherapy Protocols ,Neoplasm Metastasis ,Child ,Osteosarcoma ,Tumor ,Ifosfamide ,biology ,Middle Aged ,Germline polymorphism ,Multidrug Resistance-Associated Protein 2 ,Neoadjuvant Therapy ,Treatment Outcome ,Italy ,030220 oncology & carcinogenesis ,Cohort ,Toxicity ,Female ,medicine.drug ,Adult ,medicine.medical_specialty ,Adolescent ,Genotype ,Bone Neoplasms ,Disease-Free Survival ,Young Adult ,03 medical and health sciences ,Genetic ,Internal medicine ,Drug response biomarker ,Biomarkers, Tumor ,medicine ,Humans ,Polymorphism ,Chemotherapy ,Polymorphism, Genetic ,business.industry ,germline polymorphisms ,Cancer ,Retrospective cohort study ,gamma-Glutamyl Hydrolase ,medicine.disease ,Personalized medicine ,Methotrexate ,030104 developmental biology ,Doxorubicin ,Case-Control Studies ,Methylenetetrahydrofolate reductase ,biology.protein ,Cisplatin ,business ,osteosarcoma ,personalized medicine ,toxicity ,Biomarkers - Abstract
// Claudia M. Hattinger 1 , Paola Biason 2 , Erika Iacoboni 1 , Sara Gagno 3 , Marilu Fanelli 1 , Elisa Tavanti 1 , Serena Vella 1 , Stefano Ferrari 4 , Andrea Roli 5 , Rossana Roncato 3 , Luciana Giodini 3 , Katia Scotlandi 1 , Piero Picci 1 , Giuseppe Toffoli 3 , Massimo Serra 1 1 Laboratory of Experimental Oncology, Orthopaedic Rizzoli Institute, Bologna, Italy 2 National Institute of Health and Medical Research (INSERM), Unity 892, University of Medicine of Angers, Angers, France 3 Experimental and Clinical Pharmacology Unit, National Cancer Institute, Aviano, Italy 4 Chemotherapy Ward of Muscoloskeletal Tumours, Orthopaedic Rizzoli Institute, Bologna, Italy 5 Department of Computer Science and Engineering (DISI), University of Bologna, Cesena, Italy Correspondence to: Massimo Serra, email: massimo.serra@ior.it Keywords: osteosarcoma, germline polymorphisms, toxicity, drug response biomarkers, personalized medicine Received: June 01, 2016 Accepted: July 29, 2016 Published: August 22, 2016 ABSTRACT This study aimed to identify associations between germline polymorphisms and risk of high-grade osteosarcoma (HGOS) development, event-free survival (EFS) and toxicity in HGOS patients treated with neo-adjuvant chemotherapy and surgery. Germline polymorphisms of 31 genes known to be relevant for transport or metabolism of all four drugs used in HGOS chemotherapy (methotrexate, doxorubicin, cisplatin and ifosfamide) were genotyped in 196 patients with HGOS and in 470 healthy age and gender-matched controls. Of these 196 HGOS patients, a homogeneously treated group of 126 patients was considered for survival analyses (survival cohort). For 57 of these, treatment-related toxicity data were available (toxicity cohort). Eleven polymorphisms were associated with increased risk of developing HGOS ( p < 0.05). The distribution of polymorphisms in patients was characterized by a higher Shannon entropy. In the survival cohort ( n = 126, median follow-up = 126 months), genotypes of ABCC2_1249A/G, GGH_452T/C, TP53_IVS2+38G/C and CYP2B6*6 were associated with EFS ( p < 0.05). In the toxicity cohort ( n = 57), genotypes of ABCB1_1236T/C, ABCC2_1249A/G, ABCC2_3972A/G, ERCC1_8092T/G, XPD_23591A/G, XRCC3_18067T/C, MTHFR_1298A/C and GGH_16T/C were associated with elevated risk for toxicity development ( p < 0.05). The results obtained in this retrospective study indicate that the aforementioned germline polymorphisms significantly impact on the risk of HGOS development, EFS and the occurrence of chemotherapy-related toxicity. These findings should be prospectively validated with the aim of optimizing and tailoring HGOS treatment in the near future.
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- 2016
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25. PIWI-interacting RNA (piRNA) signatures in human cardiac progenitor cells
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Antonio Lo Nigro, Serena Vella, Alessia Gallo, Pier Giulio Conaldi, Daniele Galvagno, Giuseppe Maria Raffa, and Michele Pilato
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0301 basic medicine ,Transposable element ,endocrine system ,Cell ,Piwi-interacting RNA ,piRNA ,030204 cardiovascular system & hematology ,Biology ,Biochemistry ,Cardiospheres ,03 medical and health sciences ,0302 clinical medicine ,Cardiac progenitor cells ,medicine ,Humans ,Epigenetics ,RNA, Small Interfering ,Oligonucleotide Array Sequence Analysis ,Regulation of gene expression ,Genetics ,urogenital system ,Cell Biology ,Fibroblasts ,Cardiosphere-derived cells ,Fold change ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Gene Expression Regulation ,DNA microarray ,Stem cell ,Myoblasts, Cardiac - Abstract
Cardiac progenitors, such as cardiospheres and cardiosphere-derived cells, represent an attractive cell source for cardiac regeneration. The PIWI-interacting RNAs, piRNAs, are an intriguing class of small non-coding RNAs, implicated in the regulation of epigenetic state, maintenance of genomic integrity and stem cell functions. Although non-coding RNAs are an exploiting field in cardiovascular research, the piRNA signatures of cardiac progenitors has not been evaluated yet.We profiled, through microarrays, 15,311 piRNAs expressed in cardiospheres, cardiosphere-derived cells and cardiac fibroblasts. Results showed a set of differentially expressed piRNAs (fold change ≥2, p
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- 2016
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26. Lung resident mesenchymal cells isolated from patients with the Bronchiolitis Obliterans Syndrome display a deregulated epigenetic profile
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Patrizio Vitulo, Lavinia Martino, Angelo Luca, Emanuela Cova, Federica Meloni, Salvatore Cuzzocrea, Rosa Liotta, Pier Giulio Conaldi, Serena Vella, and Alessandro Bertani
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Graft Rejection ,Male ,0301 basic medicine ,Methyltransferase ,Cellular differentiation ,Epigenesis, Genetic ,Cell Movement ,Histone code ,Myofibroblasts ,Bronchiolitis Obliterans ,Lung ,Vorinostat ,Multidisciplinary ,Cell Differentiation ,Middle Aged ,Histone Code ,Disease Progression ,Histone Methyltransferases ,Medicine ,Female ,Bronchoalveolar Lavage Fluid ,Lung Transplantation ,Adult ,Science ,Bronchiolitis obliterans ,Histone Deacetylases ,Article ,Cell Line ,Transforming Growth Factor beta1 ,03 medical and health sciences ,microRNA ,Epigenetic Profile ,medicine ,Humans ,Epigenetics ,Aged ,Cell Proliferation ,business.industry ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,medicine.disease ,Fibrosis ,Histone Deacetylase Inhibitors ,MicroRNAs ,030104 developmental biology ,Cancer research ,Transcriptome ,business - Abstract
Bronchiolitis Obliterans Syndrome is the major determinant of the graft function loss after lung transplantation, but its pathogenesis is still incompletely understood and currently available therapeutic strategies are poorly effective. A deeper understanding of its pathogenic mechanisms is crucial for the development of new strategies to prevent and treat this devastating complication. In this study, we focused on the mesenchymal stromal cells, recently recognized as BOS key effectors, and our primary aim was to identify their epigenetic determinants, such as histone modifications and non-coding RNA regulation, which could contribute to their differentiation in myofibroblasts. Interestingly, we identified a deregulated expression of histone deacetylases and methyltransferases, and a microRNA-epigenetic regulatory network, which could represent novel targets for anti-fibrotic therapy. We validated our results in vitro, in a cell model of fibrogenesis, confirming the epigenetic involvement in this process and paving the way for a new application for epigenetic drugs.
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- 2018
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27. Advances in emerging drugs for osteosarcoma
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Massimo Serra, Claudia Maria Hattinger, Piero Picci, Elisa Tavanti, Serena Vella, Stefano Ferrari, and Marilù Fanelli
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Oncology ,Treatment response ,Pathology ,medicine.medical_specialty ,Antineoplastic Agents ,Bone Neoplasms ,Clinical investigation ,Surgical removal ,Internal medicine ,medicine ,Recurrent disease ,Animals ,Humans ,Pharmacology (medical) ,Pharmacology ,Osteosarcoma ,Postoperative chemotherapy ,business.industry ,Patient Selection ,Prognosis ,medicine.disease ,Combined Modality Therapy ,Drug Design ,Pharmacogenomics ,business ,Pharmacogenetics - Abstract
Osteosarcoma (OS), the most common primary malignant bone tumor, is currently treated with pre- and postoperative chemotherapy in association with the surgical removal of the tumor. Conventional treatments allow to cure about 60 - 65% of patients with primary tumors and only 20 - 25% of patients with recurrent disease. New treatment approaches and drugs are therefore highly warranted to improve prognosis.This review focuses on the therapeutic approaches that are under development or clinical evaluation in OS. Information was obtained from different and continuously updated data bases, as well as from literature searches, in which particular relevance was given to reports and reviews on new targeted therapies under clinical investigation in high-grade OS.OS is a heterogeneous tumor, with a great variability in treatment response between patients. It is therefore unlikely that a single therapeutic tool will be uniformly successful for all OS patients. This claims for the validation of new treatment approaches together with biologic/(pharmaco)genetic markers, which may select the most appropriate subgroup of patients for each treatment approach. Since some promising novel agents and treatment strategies are currently tested in Phase I/II/III clinical trials, we may hope that new therapies with superior efficacy and safety profiles will be identified in the next few years.
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- 2015
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28. Doxorubicin-Resistant Osteosarcoma: novel therapeutic approaches in sight?
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Piero Picci, Serena Vella, Elisa Tavanti, Claudia Maria Hattinger, Massimo Serra, Chiara Riganti, and Marilù Fanelli
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0301 basic medicine ,Cancer Research ,Bone Neoplasms ,Drug resistance ,doxorubicin ,03 medical and health sciences ,0302 clinical medicine ,osteosarcoma ,Antineoplastic Combined Chemotherapy Protocols ,Humans ,Medicine ,Doxorubicin ,Doxorubicin resistant ,Antibiotics, Antineoplastic ,drug resistance ,Drug Substitution ,business.industry ,General Medicine ,medicine.disease ,Treatment Outcome ,030104 developmental biology ,Oncology ,Drug Resistance, Neoplasm ,030220 oncology & carcinogenesis ,Cancer research ,Osteosarcoma ,business ,medicine.drug - Published
- 2017
29. Pharmacogenomics of second-line drugs used for treatment of unresponsive or relapsed osteosarcoma patients
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Piero Picci, Serena Vella, Elisa Tavanti, Massimo Serra, Claudia Maria Hattinger, and Marilù Fanelli
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0301 basic medicine ,Oncology ,medicine.medical_specialty ,medicine.medical_treatment ,Bone Neoplasms ,Drug resistance ,Review ,Pharmacology ,03 medical and health sciences ,0302 clinical medicine ,Second line ,Internal medicine ,Antineoplastic Combined Chemotherapy Protocols ,Genetics ,medicine ,Humans ,Relapsed Osteosarcoma ,Chemotherapy ,Osteosarcoma ,business.industry ,medicine.disease ,030104 developmental biology ,Pharmacogenetics ,030220 oncology & carcinogenesis ,Pharmacogenomics ,Molecular Medicine ,Neoplasm Recurrence, Local ,business - Abstract
Second-line treatment of high-grade osteosarcoma (HGOS) patients is based on different approaches and chemotherapy protocols, which are not yet standardized. Although several drugs have been used in HGOS second-line protocols, none of them has provided fully satisfactory results and the role of rescue chemotherapy is not well defined yet. This article focuses on the drugs that have most frequently been used for second-line treatment of HGOS, highlighting the present knowledge on their mechanisms of action and resistance and on gene polymorphisms with possible impact on treatment sensitivity or toxicity. In the near future, validation of the so far identified candidate genetic biomarkers may constitute the basis for tailoring treatment by taking the patients’ genetic background into account.
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- 2016
30. Pharmacogenomics of genes involved in antifolate drug response and toxicity in osteosarcoma
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Marilù Fanelli, Claudia Maria Hattinger, Serena Vella, Elisa Tavanti, Piero Picci, and Massimo Serra
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0301 basic medicine ,Antimetabolites, Antineoplastic ,Bone Neoplasms ,Pemetrexed ,Pharmacology ,Toxicology ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,medicine ,Humans ,Osteosarcoma ,business.industry ,General Medicine ,medicine.disease ,030104 developmental biology ,Trimetrexate ,Methotrexate ,chemistry ,Pharmacogenetics ,030220 oncology & carcinogenesis ,Pharmacogenomics ,Toxicity ,Antifolate ,Folic Acid Antagonists ,Neoplasm Grading ,business ,medicine.drug - Abstract
Antifolates are structural analogs of folates, which have been used as antitumor drugs for more than 60 years. The antifolate drug most commonly used for treating human tumors is methotrexate (MTX), which is utilized widely in first-line treatment protocols of high-grade osteosarcoma (HGOS). In addition to MTX, two other antifolates, trimetrexate and pemetrexed, have been tested in clinical settings for second-line treatment of recurrent HGOS with patients unfortunately showing modest activity. Areas covered: There is clinical evidence which suggsest that, like other chemotherapeutic agents, not all HGOS patients are equally responsive to antifolates and do not have the same susceptibility to experience adverse drug-related toxicities. Here, we summarize the pharmacogenomic information reported so far for genes involved in antifolate metabolism and transport and in MTX-related toxicity in HGOS patients. Expert opinion: Identification and validation of genetic biomarkers that significantly impact clinical antifolate treatment response and related toxicity may provide the basis for a future treatment modulation based on the pharmacogenetic and pharmacogenomic features of HGOS patients.
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- 2016
31. Global profiling of viral and cellular non-coding RNAs in Epstein-Barr virus-induced lymphoblastoid cell lines and released exosome cargos
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Serena Vella, Monica Miele, Alessia Gallo, Mariangela Di Bella, Pier Giulio Conaldi, Silvia Bosi, Francesca Timoneri, and Marco Sciveres
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0301 basic medicine ,Cancer Research ,Epstein-Barr Virus Infections ,Lymphoma ,Biology ,medicine.disease_cause ,Exosomes ,Exosome ,03 medical and health sciences ,hemic and lymphatic diseases ,Cell Line, Tumor ,microRNA ,otorhinolaryngologic diseases ,medicine ,Humans ,Tumor microenvironment ,RNA ,Non-coding RNA ,Virology ,Epstein–Barr virus ,Long non-coding RNA ,Microvesicles ,Cell biology ,030104 developmental biology ,Oncology ,RNA, Long Noncoding - Abstract
The human EBV-transformed lymphoblastoid cell line (LCL), obtained by infecting peripheral blood monocular cells with Epstein-Barr Virus, has been extensively used for human genetic, pharmacogenomic, and immunologic studies. Recently, the role of exosomes has also been indicated as crucial in the crosstalk between EBV and the host microenvironment. Because the role that the LCL and LCL exosomal cargo might play in maintaining persistent infection, and since little is known regarding the non-coding RNAs of LCL, the aim of our work was the comprehensive characterization of this class of RNA, cellular and viral miRNAs, and cellular lncRNAs, in LCL compared with PBMC derived from the same donors. In this study, we have demonstrated, for the first time, that all the viral miRNAs expressed by LCL are also packaged in the exosomes, and we found that two miRNAs, ebv-miR-BART3 and ebv-miR-BHRF1-1, are more abundant in the exosomes, suggesting a microvescicular viral microRNA transfer. In addition, lncRNA profiling revealed that LCLs were enriched in lncRNA H19 and H19 antisense, and released these through exosomes, suggesting a leading role in the regulation of the tumor microenvironment.
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- 2016
32. Targeting CDKs with Roscovitine Increases Sensitivity to DNA Damaging Drugs of Human Osteosarcoma Cells
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Massimo Serra, Piero Picci, Claudia Maria Hattinger, Marilù Fanelli, Jan Koster, Rogier Versteeg, Serena Vella, Elisa Tavanti, Cancer Center Amsterdam, Oncogenomics, and Amsterdam Gastroenterology Endocrinology Metabolism
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0301 basic medicine ,Cancer Treatment ,lcsh:Medicine ,Apoptosis ,Biochemistry ,0302 clinical medicine ,Medicine and Health Sciences ,Drug Interactions ,Small interfering RNAs ,Molecular Targeted Therapy ,Cell Cycle and Cell Division ,lcsh:Science ,Osteosarcoma ,Multidisciplinary ,biology ,Cell Death ,Pharmaceutics ,Cell Cycle ,Drug Synergism ,Cell cycle ,Gene Expression Regulation, Neoplastic ,Nucleic acids ,Oncology ,Cell Processes ,030220 oncology & carcinogenesis ,medicine.drug ,Research Article ,DNA damage ,DNA repair ,Antineoplastic Agents ,03 medical and health sciences ,Drug Therapy ,Cyclin-dependent kinase ,Cell Line, Tumor ,medicine ,Roscovitine ,Genetics ,Humans ,Doxorubicin ,Gene Silencing ,Non-coding RNA ,Cell Proliferation ,Cisplatin ,Pharmacology ,Biology and life sciences ,Cell growth ,business.industry ,Cyclin-dependent kinase 2 ,lcsh:R ,Cyclin-Dependent Kinase 2 ,DNA ,Cell Biology ,Gene regulation ,030104 developmental biology ,Purines ,Cancer research ,biology.protein ,RNA ,lcsh:Q ,Gene expression ,business - Abstract
Cyclin-dependent kinase 2 (CDK2) has been reported to be essential for cell proliferation in several human tumours and it has been suggested as an appropriate target to be considered in order to enhance the efficacy of treatment regimens based on the use of DNA damaging drugs. We evaluated the clinical impact of CDK2 overexpression on a series of 21 high-grade osteosarcoma (OS) samples profiled by using cDNA microarrays. We also assessed the in vitro efficacy of the CDKs inhibitor roscovitine in a panel of drug-sensitive and drug-resistant human OS cell lines. OS tumour samples showed an inherent overexpression of CDK2, and high expression levels at diagnosis of this kinase appeared to negatively impact on clinical outcome. CDK2 expression also proved to be relevant for in vitro OS cells growth. These findings indicated CDK2 as a promising candidate therapeutic marker for OS and therefore we assessed the efficacy of the CDKs-inhibitor roscovitine in both drug-sensitive and -resistant OS cell lines. All cell lines resulted to be responsive to roscovitine, which was also able to increase the activity of cisplatin and doxorubicin, the two most active DNA damaging drugs used in OS chemotherapy. Our results indicated that combined treatment with conventional OS chemotherapeutic drugs and roscovitine may represent a new candidate intervention approach, which may be considered to enhance tumour cell sensitivity to DNA damaging drugs.
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- 2016
33. Cystatin S-a candidate biomarker for severity of submandibular gland involvement in Sjögren's syndrome
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Chiara Baldini, Daniela Martini, Serena Vella, Marta Mosca, Enza Polizzi, Francesca Sernissi, Nicoletta Luciano, Antonella Cecchettini, Alessia Gallo, and Pier Giulio Conaldi
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0301 basic medicine ,medicine.medical_specialty ,Submandibular Gland ,Sjögren’s syndrome ,cystatin S ,miRNA ,salivary proteomics ,03 medical and health sciences ,0302 clinical medicine ,stomatognathic system ,Rheumatology ,Downregulation and upregulation ,Western blot ,Internal medicine ,microRNA ,Submandibular Gland Diseases ,medicine ,Humans ,Pharmacology (medical) ,Saliva ,030203 arthritis & rheumatology ,Salivary gland ,medicine.diagnostic_test ,business.industry ,Middle Aged ,medicine.disease ,Submandibular gland ,Peeling skin syndrome ,MicroRNAs ,030104 developmental biology ,Real-time polymerase chain reaction ,medicine.anatomical_structure ,Endocrinology ,Sjogren's Syndrome ,Case-Control Studies ,Biomarker (medicine) ,Salivary Cystatins ,Female ,business ,Biomarkers - Abstract
Objectives Salivary cystatin S is a defence protein mainly produced by submandibular glands and involved in innate oral immunity. This study aimed to verify whether cystatin S was diversely expressed in different disease subsets of primary Sjogren's syndrome (pSS) patients, defined on the basis of salivary flow [unstimulated salivary flow rate (USFR)], minor salivary gland (MSG) focus score and submandibular gland ultrasonography abnormalities. We also evaluated miR-126 and miR-335-5p expression in MSG biopsies to verify whether an aberrant regulation of cystatin S at the glandular level may influence its salivary expression. Methods Forty pSS patients and 20 sex- and age-matched healthy volunteers were included. Salivary cystatin S levels were assessed by western blot analysis using a stain-free technology. The expression of miR-126, miR-335-5p and cystatin S was assessed by quantitative PCR in 15 MSG biopsies differing for USFR and MSG focus score. Results We found that salivary cystatin S was significantly decreased in pSS patients vs healthy volunteers ( P = 0.000), especially in those with hyposalivation. A positive correlation was observed between cystatin S and USFR ( r = 0.75, P = 0.01). Salivary cystatin S was also significantly reduced in patients with a submandibular gland ultrasonography score ⩾2. The expression levels of miR-126 and miR-335-5P increased in inverse proportion with USFR. The mRNA of cystatin S did not change significantly, suggesting post-transcriptional regulation. Conclusion Cystatin S emerged as a promising biomarker for pSS, strongly correlated with glandular dysfunction. An upregulation of miR-126 and miR-335-5P might be implicated in its expression.
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- 2016
34. NDM29, a RNA polymerase III-dependent non coding RNA, promotes amyloidogenic processing of APP and amyloid β secretion
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Ranieri Cancedda, Serena Vella, Tullio Florio, Claudio Russo, Mario Nizzari, Aldo Pagano, Eleonora Ciarlo, and Sara Massone
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Small RNA ,RNA polymerase III ,RNA, Untranslated ,BACE1-AS ,Models, Biological ,chemistry.chemical_compound ,Neuroblastoma ,Alzheimer Disease ,RNA polymerase ,Amyloid precursor protein ,Humans ,Non-coding RNA ,Molecular Biology ,Cerebral Cortex ,Inflammation ,Neurons ,Amyloid beta-Peptides ,biology ,P3 peptide ,RNA ,Cell Differentiation ,Beta amyloid ,Cell Biology ,Alzheimer's disease ,Molecular biology ,Phenotype ,RNA polymerase III, Alzheimer's disease, Beta amyloid, Neuroblastoma, Non-coding RNA ,chemistry ,Postmortem Changes ,biology.protein ,Protein Processing, Post-Translational - Abstract
Neuroblastoma Differentiation Marker 29 (NDM29) is a RNA polymerase (pol) III-transcribed non-coding (nc) RNA whose synthesis drives neuroblastoma (NB) cell differentiation to a nonmalignant neuron-like phenotype. Since in this process a complex pattern of molecular changes is associated to plasma membrane protein repertoire we hypothesized that the expression of NDM29 might influence also key players of neurodegenerative pathways. In this work we show that the NDM29-dependent cell maturation induces amyloid precursor protein (APP) synthesis, leading to the increase of amyloid β peptide (Aβ) secretion and the concomitant increment of Aβ x-42/Aβ x-40 ratio. We also demonstrate that the expression of NDM29 RNA, and the consequent increase of Aβ formation, can be promoted by inflammatory stimuli (and repressed by anti-inflammatory drugs). Moreover, NDM29 expression was detected in normal human brains although an abnormal increased synthesis of this ncRNA is induced in patients affected by neurodegenerative diseases. Therefore, the complex of events triggered by NDM29 expression induces a condition that favors the formation of Aβ peptides in the extracellular space, as it may occur in Alzheimer's Disease (AD). In addition, these data unexpectedly show that a pol III-dependent small RNA can act as key regulator of brain physiology and/or pathology suggesting that a better knowledge of this portion of the human transcriptome might provide hints for neurodegeneration studies.
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- 2012
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35. Probing cytoskeleton organisation of neuroblastoma cells with single-cell force spectroscopy
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Claudio Canale, Serena Vella, Aldo Pagano, Alberto Diaspro, Marco Scotto, Paola Gavazzo, Andrea Mescola, and Massimo Vassalli
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Cell ,Force spectroscopy ,Biology ,medicine.disease ,Embryonic stem cell ,Cell biology ,Kidney cell ,Neuroblastoma cell ,medicine.anatomical_structure ,Structural Biology ,Neuroblastoma ,medicine ,Cytoskeleton organisation ,Cytoskeleton ,Molecular Biology - Abstract
Single-cell force spectroscopy is an emerging technique in the field of biomedicine because it has proved to be a unique tool to obtain mechanical and functional information on living cells, with force resolution up to single molecular bonds. This technique was applied to the study of the cytoskeleton organisation of neuroblastoma cells, a life-threatening cancer typically developing during childhood, and the results were interpreted on the basis of reference experiments on human embryonic kidney cell line. An intimate connection emerges among cellular state, cytoskeleton organisation and experimental outcome that can be potentially exploited towards a new method for cancer stadiation of neuroblastoma cells. Copyright © 2012 John Wiley & Sons, Ltd.
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- 2012
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36. Acquisition of neuron-like electrophysiological properties in neuroblastoma cells by controlled expression of NDM29 ncRNA
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Serena Vella, Carla Marchetti, Paola Gavazzo, Ranieri Cancedda, Aldo Pagano, and Mario Nizzari
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Regulation of gene expression ,Neurite ,Cellular differentiation ,Cell ,Biology ,medicine.disease ,Biochemistry ,Pediatric cancer ,Molecular biology ,Cellular and Molecular Neuroscience ,medicine.anatomical_structure ,Cell culture ,Neuroblastoma ,medicine ,Neuron - Abstract
Neuroblastoma is a pediatric cancer characterized by high malignancy and remarkable cell heterogeneity within the tumor nodules. It has been previously shown that the over-expression of a specific non-coding RNA, NDM29, reduces neuroblastoma development promoting cell differentiation. We have used neuroblastoma cells expressing NDM29 at its basal level (Mock cells) or at 5.4-fold higher levels (S1 cells) to investigate whether a functional differentiation correlates with morphological and biochemical development induced by NDM29 expression. First, analyzing the expression of specific markers we demonstrated that NDM29 expression is accompanied by a well coordinated differentiation process toward a neuron-like, rather than toward a glial-like, phenotype. Next, we defined the neuron-like traits of S1 in terms of secretion of cytokines involved in axon guidance, synapse formation and neurite outgrowth. Finally, we characterized the ionic channel apparatus of S1 cells by patch-clamp technique and compared with the Mock counterpart. S1 cells showed much higher levels of fast inactivating Na(+) current and were able to generate mature action potentials. Moreover, they developed expression of functional GABA(A) receptors on their membrane. In contrast, the two cell lines shared very similar pools of functional K(+) channels, although slight quantitative differences can be described. Our results suggest that a maturation occurs in neuroblastoma as a consequence of NDM29 expression, inducing the appearance of neuronal-like properties. In this context, S1 cells may represent a novel in vitro tool for electrophysiological and pharmacological studies of human cells of the neural lineage.
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- 2011
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37. Selection of Candidate Housekeeping Genes for Normalization in Human Postmortem Brain Samples
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Arianna Gigoni, Aldo Pagano, Ranieri Cancedda, Claudio Russo, Serena Vella, and Ilaria Penna
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Male ,Normalization (statistics) ,Cytochromes c1 ,reference genes ,Biology ,geNorm ,Catalysis ,Postmortem Changes ,Inorganic Chemistry ,lcsh:Chemistry ,Alzheimer Disease ,quantitative real-time RT-PCR ,Reference genes ,medicine ,Humans ,Physical and Theoretical Chemistry ,Molecular Biology ,Gene ,lcsh:QH301-705.5 ,Spectroscopy ,Aged ,Aged, 80 and over ,Genetics ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Expression Profiling ,Communication ,Organic Chemistry ,Brain ,NormFinder ,General Medicine ,Human brain ,Middle Aged ,Reference Standards ,medicine.disease ,Computer Science Applications ,Housekeeping gene ,Gene expression profiling ,medicine.anatomical_structure ,lcsh:Biology (General) ,lcsh:QD1-999 ,Eukaryotic Initiation Factor-4A ,Female ,Alzheimer's disease ,Alzheimer’s disease ,Algorithms - Abstract
The most frequently used technique to study the expression profile of genes involved in common neurological disorders is quantitative real-time RT-PCR, which allows the indirect detection of very low amounts of selected mRNAs in tissue samples. Expression analysis by RT-qPCR requires an appropriate normalization to the expression level of genes characterized by a stable, constitutive transcription. However, the identification of a gene transcribed at a very stable level is difficult if not impossible, since significant fluctuations of the level of mRNA synthesis often accompanies changes of cell behavior. The aim of this study is to identify the most stable genes in postmortem human brain samples of patients affected by Alzheimer’s disease (AD) suitable as reference genes. The experiments analyzed 12 commonly used reference genes in brain samples from eight individuals with AD and seven controls. After a careful analysis of the results calculated by geNorm and NormFinder algorithms, we found that CYC1 and EIF4A2 are the best reference genes. We remark on the importance of the determination of the best reference genes for each sample to be analyzed and suggest a practical combination of reference genes to be used in the analysis of human postmortem samples.
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- 2011
38. Targeting ABCB1 and ABCC1 with their Specific Inhibitor CBT-1® can Overcome Drug Resistance in Osteosarcoma
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Daryl Barnett, Claudia Maria Hattinger, Marilù Fanelli, Piero Picci, Elisa Tavanti, Francesca Michelacci, Beth Gudeman, Massimo Serra, and Serena Vella
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0301 basic medicine ,Cancer Research ,ATP Binding Cassette Transporter, Subfamily B ,medicine.medical_treatment ,Blotting, Western ,Fluorescent Antibody Technique ,Apoptosis ,Bone Neoplasms ,Drug resistance ,Pharmacology ,Vinorelbine ,Benzylisoquinolines ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Drug Discovery ,medicine ,Tumor Cells, Cultured ,Humans ,Doxorubicin ,Etoposide ,Cell Proliferation ,Cisplatin ,Chemotherapy ,Osteosarcoma ,Antibiotics, Antineoplastic ,business.industry ,Flow Cytometry ,Antineoplastic Agents, Phytogenic ,Drug Resistance, Multiple ,Tetrandrine ,030104 developmental biology ,Oncology ,chemistry ,Microscopy, Fluorescence ,Drug Resistance, Neoplasm ,030220 oncology & carcinogenesis ,Methotrexate ,Multidrug Resistance-Associated Proteins ,business ,medicine.drug - Abstract
Clinical treatment response achievable with conventional chemotherapy in high-grade osteosarcoma (OS) is severely limited by the presence of intrinsic or acquired drug resistance, which in previous studies has been mainly addressed for overexpression of ABCB1 (MDR1/P-glycoprotein). This study was aimed to estimate the impact on OS drug resistance of a group of ATP binding cassette (ABC) transporters, which in other human tumors have been associated with unresponsiveness to the drugs that represent the backbone of multidrug treatment regimens for OS (doxorubicin, methotrexate, cisplatin). By using a group of 6 drug-sensitive and 20 drug-resistant human OS cell lines, the most relevant transporter which proved to be associated with the degree of drug resistance in OS cells, in addition to ABCB1, was ABCC1. We therefore evaluated the in vitro activity of the orally administrable ABCB1/ABCC1 inhibitor CBT-1(®) (Tetrandrine, NSC-77037). We found that in our OS cell lines this agent was able to revert the ABCB1/ABCC1-mediated resistance against doxorubicin, as well as against the drugs used in second-line OS treatments that are substrates of these transporters (taxotere, etoposide, vinorelbine). Our findings indicated that inhibiting ABCB1 and ABCC1 with CBT-1(®), used in association with conventional chemotherapeutic drugs, may become an interesting new therapeutic option for unresponsive or relapsed OS patients.
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- 2015
39. Targeting polo-like kinase 1 by NMS-P937 in osteosarcoma cell lines inhibits tumor cell growth and partially overcomes drug resistance
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Francesca Michelacci, Elisa Tavanti, Barbara Valsasina, Serena Vella, Rogier Versteeg, Massimo Serra, Beth Gudeman, Claudia Maria Hattinger, Marilù Fanelli, Piero Picci, Valeria Sero, Cancer Center Amsterdam, and Oncogenomics
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ATP Binding Cassette Transporter, Subfamily B ,Antineoplastic Agents ,Apoptosis ,Bone Neoplasms ,Cell Cycle Proteins ,Polo-like kinase ,Pharmacology ,Biology ,Protein Serine-Threonine Kinases ,PLK1 ,Cell Line, Tumor ,Proto-Oncogene Proteins ,medicine ,Gene silencing ,Humans ,Pharmacology (medical) ,Doxorubicin ,Drug Interactions ,RNA, Messenger ,RNA, Small Interfering ,Clonogenic assay ,Protein Kinase Inhibitors ,Cell Proliferation ,Osteosarcoma ,Gene Expression Profiling ,Cell Cycle ,Cell cycle ,medicine.disease ,Oncology ,Cell culture ,Drug Resistance, Neoplasm ,Quinazolines ,Pyrazoles ,medicine.drug - Abstract
Background Polo-like kinase 1 (PLK1) has emerged as a prognostic factor in various neoplasms, but only scarce data have been reported for high-grade osteosarcoma (OS). In this study, we assessed PLK1 expression and the efficacy of PLK1 inhibitor NMS-P937 in OS. Methods PLK1 expression was assessed on 21 OS clinical samples and on a panel of human OS cell lines. In vitro efficacy of NMS-P937 was evaluated on nine drug-sensitive and six drug-resistant human OS cell lines, either as single agent or in combination with the drugs used in chemotherapy for OS. Results PLK1 expression was higher in OS clinical samples and cell lines compared to normal human tissue. A higher PLK1 expression at diagnosis appeared to be associated with an unfavourable clinical outcome. PLK1 silencing produced growth inhibition, cell cycle retardation and apoptosis induction in human OS cell lines. NMS-P937 proved to be highly active in both drug-sensitive and drug-resistant cell lines, with the only exception of ABCB1-overexpressing, Doxorubicin (DX)-resistant variants. However, in these cells, the association of NMS-P937 with DX was able to revert DX-resistance by negatively interfering with ABCB1 transport activity. NMS-P937 was also able to decrease clonogenic and migration ability of human OS cell lines. Conclusion PLK1 can be proposed as a new candidate target for OS. Targeting PLK1 in OS with NMS-P937 in association with conventional chemotherapeutic drugs may be a new interesting therapeutic option, since this approach has proved to be active against drug resistant cells.
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- 2014
40. Hyper-activation of notch3 amplifies the proliferative potential of rhabdomyosarcoma cells
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DE SALVO, Maria, Lavinia, Raimondi, Serena, Vella, Adesso, Laura, Roberta, Ciarapica, Federica, Verginelli, Antonio, Pannuti, Citti, Arianna, Renata, Boldrini, Milano, Giuseppe M., Antonella, Cacchione, Andrea, Ferrari, Paola, Collini, Angelo, Rosolen, Gianni, Bisogno, Rita, Alaggio, Alessandro, Inserra, Mattia, Locatelli, Stefano, Stifani, Screpanti, Isabella, Lucio, Miele, Locatelli, Franco, Rossella, Rota, and Alaggio, Rita
- Subjects
Pathology ,Oncogene Proteins, Fusion ,genetic structures ,Tumor Physiology ,Cellular differentiation ,NF-KAPPA-B ,lcsh:Medicine ,Gene Expression ,Pediatrics ,Myoblast fusion ,Basic Cancer Research ,Medicine and Health Sciences ,Paired Box Transcription Factors ,Rhabdomyosarcoma, Embryonal ,Receptor, Notch1 ,HES1 ,lcsh:Science ,Rhabdomyosarcoma ,Receptor, Notch3 ,TRANSGENIC MICE ,Multidisciplinary ,Receptors, Notch ,Myogenesis ,Sarcomas ,Animal Models ,MUSCLE ,musculoskeletal system ,SOLID TUMORS ,Cell biology ,Gene Expression Regulation, Neoplastic ,DIFFERENTIATION ,N/A ,Settore MED/38 - PEDIATRIA GENERALE E SPECIALISTICA ,Oncology ,SIGNALING PATHWAY ,Signal Transduction ,Research Article ,EXPRESSION ,medicine.medical_specialty ,INHIBITION ,Notch signaling pathway ,Mouse Models ,Biology ,Research and Analysis Methods ,Model Organisms ,ALVEOLAR RHABDOMYOSARCOMA ,SATELLITE CELLS ,Cell Line, Tumor ,Genetics ,medicine ,Humans ,Rhabdomyosarcoma, Alveolar ,Cell Proliferation ,Cell growth ,lcsh:R ,Biology and Life Sciences ,Cancers and Neoplasms ,medicine.disease ,Ki-67 Antigen ,Pediatric Oncology ,Cell culture ,Settore MED/20 ,lcsh:Q ,human activities - Abstract
Rhabdomyosarcoma (RMS) is a pediatric myogenic-derived soft tissue sarcoma that includes two major histopathological subtypes: embryonal and alveolar. The majority of alveolar RMS expresses PAX3-FOXO1 fusion oncoprotein, associated with the worst prognosis. RMS cells show myogenic markers expression but are unable to terminally differentiate. The Notch signaling pathway is a master player during myogenesis, with Notch1 activation sustaining myoblast expansion and Notch3 activation inhibiting myoblast fusion and differentiation. Accordingly, Notch1 signaling is up-regulated and activated in embryonal RMS samples and supports the proliferation of tumor cells. However, it is unable to control their differentiation properties. We previously reported that Notch3 is activated in RMS cell lines, of both alveolar and embryonal subtype, and acts by inhibiting differentiation. Moreover, Notch3 depletion reduces PAX3-FOXO1 alveolar RMS tumor growth in vivo. However, whether Notch3 activation also sustains the proliferation of RMS cells remained unclear. To address this question, we forced the expression of the activated form of Notch3, Notch3IC, in the RH30 and RH41 PAX3-FOXO1-positive alveolar and in the RD embryonal RMS cell lines and studied the proliferation of these cells. We show that, in all three cell lines tested, Notch3IC over-expression stimulates in vitro cell proliferation and prevents the effects of pharmacological Notch inhibition. Furthermore, Notch3IC further increases RH30 cell growth in vivo. Interestingly, knockdown of Notch canonical ligands JAG1 or DLL1 in RMS cell lines decreases Notch3 activity and reduces cell proliferation. Finally, the expression of Notch3IC and its target gene HES1 correlates with that of the proliferative marker Ki67 in a small cohort of primary PAX-FOXO1 alveolar RMS samples. These results strongly suggest that high levels of Notch3 activation increase the proliferative potential of RMS cells.
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- 2014
41. EZH2 down-regulation exacerbates lipid accumulation and inflammation in in vitro and in vivo NAFLD
- Author
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Annalisa Crudele, Serena Vella, Franco Locatelli, Victor E. Marquez, Sara Ceccarelli, Stefania Gaspari, Daniela Gnani, Valerio Nobili, Anna Alisi, Rossella Rota, and Cristiano De Stefanis
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Adenosine ,Messenger ,Palmitic Acid ,lcsh:Chemistry ,Histones ,Rats, Sprague-Dawley ,Non-alcoholic Fatty Liver Disease ,Transforming Growth Factor beta ,lcsh:QH301-705.5 ,Spectroscopy ,DZNep ,EZH2 ,microRNAs ,NAFLD ,Animals ,Disease Models, Animal ,Enhancer of Zeste Homolog 2 Protein ,Fatty Liver ,Hep G2 Cells ,Humans ,MicroRNAs ,Oleic Acid ,Polycomb Repressive Complex 2 ,RNA, Messenger ,Rats ,Tumor Necrosis Factor-alpha ,Down-Regulation ,Catalysis ,Molecular Biology ,Computer Science Applications1707 Computer Vision and Pattern Recognition ,Physical and Theoretical Chemistry ,Organic Chemistry ,Inorganic Chemistry ,Fatty liver ,General Medicine ,Computer Science Applications ,Biochemistry ,Settore MED/38 - PEDIATRIA GENERALE E SPECIALISTICA ,Tumor necrosis factor alpha ,medicine.symptom ,Inflammation ,macromolecular substances ,Biology ,Article ,Histone H3 ,microRNA ,medicine ,Epigenetics ,Animal ,medicine.disease ,lcsh:Biology (General) ,lcsh:QD1-999 ,Disease Models ,Cancer research ,RNA ,Sprague-Dawley ,Steatosis - Abstract
Non-alcoholic fatty liver disease (NAFLD) is one of the most prevalent, chronic liver diseases, worldwide. It is a multifactorial disease caused by complex interactions between genetic, epigenetic and environmental factors. Recently, several microRNAs, some of which epigenetically regulated, have been found to be up- and/or down-regulated during NAFLD development. However, in NAFLD, the essential role of the Polycomb Group protein Enhancer of Zeste Homolog 2 (EZH2), which controls the epigenetic silencing of specific genes and/or microRNAs by trimethylating Lys27 on histone H3, still remains unknown. In this study, we demonstrate that the nuclear expression/activity of the EZH2 protein is down-regulated both in livers from NAFLD rats and in the free fatty acid-treated HepG2. The drop in EZH2 is inversely correlated with: (i) lipid accumulation; (ii) the expression of pro-inflammatory markers including TNF-α and TGF-β; and (iii) the expression of miR-200b and miR-155. Consistently, the pharmacological inhibition of EZH2 by 3-Deazaneplanocin A (DZNep) significantly reduces EZH2 expression/activity, while it increases lipid accumulation, inflammatory molecules and microRNAs. In conclusion, the results of this study suggest that the defective activity of EZH2 can enhance the NAFLD development by favouring steatosis and the de-repression of the inflammatory genes and that of specific microRNAs.
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- 2013
42. Differential toxicity, conformation and morphology of typical initial aggregation states of Aβ1-42 and Aβpy3-42 beta-amyloids
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Cristina D'Arrigo, Tullio Florio, Serena Vella, Denise Galante, Alessandro Corsaro, Massimo Vassalli, Aldo Pagano, Francesca Sbrana, and Angelo Perico
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Circular dichroism ,Time Factors ,Proteolysis ,Peptide ,Biochemistry ,Oligomer ,Protein Structure, Secondary ,chemistry.chemical_compound ,Protein structure ,Cell Line, Tumor ,medicine ,Humans ,Protein Structure, Quaternary ,Chromatography, High Pressure Liquid ,chemistry.chemical_classification ,Amyloid beta-Peptides ,medicine.diagnostic_test ,Cell Death ,Circular Dichroism ,Neurotoxicity ,Fast protein liquid chromatography ,Cell Biology ,medicine.disease ,Random coil ,Endocytosis ,Peptide Fragments ,chemistry ,Microscopy, Fluorescence - Abstract
Among the different species of water-soluble β-peptides (Aβ1-42, Aβ1-40 and N-terminal truncated Aβ-peptides), Aβpy3-42 is thought to play a relevant role in Alzheimer's pathogenesis due to its abundance, resistance to proteolysis, fast aggregation kinetics, dynamic structure and high neurotoxicity. To evaluate the specific structural characteristics and neurotoxicity of Aβpy3-42, we separated different aggregation states of Aβ1-42 and Aβpy3-42 using fast protein liquid chromatography, isolating in both cases three peaks that corresponded to sa (small), ma (medium) and la (large) aggregates. Conformational analysis, by circular dichroism showed a prevailing random coil conformation for sa and ma, and typical β-sheet conformation for la. AFM and TEM show differential structural features between the three aggregates of a given β-peptide and among the aggregate of the two β-peptides. The potential toxic effects of the different aggregates were evaluated using human neuroblastoma SH-SY5Y cells in the MTT reduction, in the xCELLigence System, and in the Annexin V binding experiments. In the case of Aβ1-42 the most toxic aggregate is la, while in the case of Aβpy3-42 both sa and la are equally toxic. Aβ aggregates were found to be internalized in the cells, as estimated by confocal immunofluorescence microscopy, with a higher effect observed for Aβpy3-42, showing a good correlation with the toxic effects. Together these experiments allowed the discrimination of the intermediate states more responsible of oligomer toxicity, providing new insights on the correlation between the aggregation process and the toxicity and confirming the peculiar role in the pathogenesis of Alzheimer disease of Aβpy3-42 peptide.
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- 2012
43. Dichloroacetate inhibits neuroblastoma growth by specifically acting against malignant undifferentiated cells
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Ranieri Cancedda, Matteo Conti, Aldo Pagano, Roberta Tasso, and Serena Vella
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Cancer Research ,medicine.medical_specialty ,Cell type ,Tumor cells ,Antineoplastic Agents ,Apoptosis ,Mice, SCID ,Biology ,Mitochondrion ,Mice ,Neuroblastoma ,Mice, Inbred NOD ,Internal medicine ,medicine ,Tumor Cells, Cultured ,Animals ,Humans ,Cell Proliferation ,Membrane potential ,Membrane Potential, Mitochondrial ,Dichloroacetic Acid ,Cell Differentiation ,Metabolism ,medicine.disease ,Phenotype ,Mitochondria ,Endocrinology ,Oncology ,Cancer cell ,Cancer research - Abstract
The small, water soluble molecule Dichloroacetate (DCA) is recently arousing lively interests in the field of cancer therapy for it has been shown to be able to inhibit the growth of human tumors acting specifically on the mitochondria of cancer cells without perturbing the physiology of nonmalignant cells. Neuroblastoma was one of the tumor types on which DCA was considered ineffective as it is composed of cells with few recognized mitochondrial anomalies. Neuroblastoma, however, is composed of different cell types in terms of metabolism, phenotype and malignant potential. Despite the above prediction, in this work, we show that (i) DCA exhibits an unexpected anticancer effect on NB tumor cells and (ii) this effect is selectively directed to very malignant NB cells, whereas the more differentiated/less malignant NB cells are refractory to DCA treatment. This result supports the need of a detailed investigation of DCA anticancer properties against this tumor type with the final aim of its possible use as therapeutic agent.
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- 2011
44. Acquisition of neuron-like electrophysiological properties in neuroblastoma cells by controlled expression of NDM29 ncRNA
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Paola, Gavazzo, Serena, Vella, Carla, Marchetti, Mario, Nizzari, Ranieri, Cancedda, and Aldo, Pagano
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Gene Expression Regulation, Neoplastic ,Neurons ,Neuroblastoma ,RNA, Untranslated ,Cell Line, Tumor ,Neurogenesis ,Cell Membrane ,Cytokines ,Humans ,Cell Differentiation ,Nerve Tissue Proteins ,Biomarkers - Abstract
Neuroblastoma is a pediatric cancer characterized by high malignancy and remarkable cell heterogeneity within the tumor nodules. It has been previously shown that the over-expression of a specific non-coding RNA, NDM29, reduces neuroblastoma development promoting cell differentiation. We have used neuroblastoma cells expressing NDM29 at its basal level (Mock cells) or at 5.4-fold higher levels (S1 cells) to investigate whether a functional differentiation correlates with morphological and biochemical development induced by NDM29 expression. First, analyzing the expression of specific markers we demonstrated that NDM29 expression is accompanied by a well coordinated differentiation process toward a neuron-like, rather than toward a glial-like, phenotype. Next, we defined the neuron-like traits of S1 in terms of secretion of cytokines involved in axon guidance, synapse formation and neurite outgrowth. Finally, we characterized the ionic channel apparatus of S1 cells by patch-clamp technique and compared with the Mock counterpart. S1 cells showed much higher levels of fast inactivating Na(+) current and were able to generate mature action potentials. Moreover, they developed expression of functional GABA(A) receptors on their membrane. In contrast, the two cell lines shared very similar pools of functional K(+) channels, although slight quantitative differences can be described. Our results suggest that a maturation occurs in neuroblastoma as a consequence of NDM29 expression, inducing the appearance of neuronal-like properties. In this context, S1 cells may represent a novel in vitro tool for electrophysiological and pharmacological studies of human cells of the neural lineage.
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- 2011
45. Human PrP90-231-induced cell death is associated with intracellular accumulation of insoluble and protease-resistant macroaggregates and lysosomal dysfunction
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Tullio Florio, Valentina Villa, Serena Vella, Aldo Pagano, Stefano Thellung, Alessandro Corsaro, and Alessandro Simi
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Cancer Research ,Programmed cell death ,Immunology ,Cathepsin D ,Biology ,cell internalization ,Cellular and Molecular Neuroscience ,chemistry.chemical_compound ,Cytosol ,Lysosome ,Cell Line, Tumor ,medicine ,Humans ,PrPC Proteins ,proteinase K resistence ,lysoome dysfunction ,Lucifer yellow ,Cell Death ,Vesicle ,apoptosis ,Cell Biology ,prion diseases ,Cell biology ,medicine.anatomical_structure ,chemistry ,Solubility ,Apoptosis ,prion protein ,Original Article ,Endopeptidase K ,Lysosomes ,Intracellular - Abstract
To define the mechanisms by which hPrP90-231 induces cell death, we analyzed its interaction with living cells and monitored its intracellular fate. Treatment of SH-SY5Y cells with fluorescein-5-isothiocyanate (FITC)-conjugated hPrP90-231 caused the accumulation of cytosolic aggregates of the prion protein fragment that increased in number and size in a time-dependent manner. The formation of large intracellular hPrP90-231 aggregates correlated with the activation of apoptosis. hPrP90-231 aggregates occurred within lysotracker-positive vesicles and induced the formation of activated cathepsin D (CD), indicating that hPrP90-231 is partitioned into the endosomal–lysosomal system structures, activating the proteolytic machinery. Remarkably, the inhibition of CD activity significantly reduced hPrP-90-231-dependent apoptosis. Internalized hPrP90-231 forms detergent-insoluble and SDS-stable aggregates, displaying partial resistance to proteolysis. By confocal microscopy analysis of lucifer yellow (LY) intracellular partition, we show that hPrP90-231 accumulation induces lysosome destabilization and loss of lysosomal membrane impermeability. In fact, although control cells evidenced a vesicular pattern of LY fluorescence (index of healthy lysosomes), hPrP90-231-treated cells showed diffuse cytosolic fluorescence, indicating LY diffusion through damaged lysosomes. In conclusion, these data indicate that exogenously added hPrP90-231 forms intralysosomal deposits having features of insoluble, protease-resistant aggregates and could trigger a lysosome-mediated apoptosis by inducing lysosome membrane permeabilization, followed by the release of hydrolytic enzymes.
- Published
- 2011
46. The inhibition of 45A ncRNA expression reduces tumor formation, affecting tumor nodules compactness and metastatic potential in neuroblastoma cells
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Delfina Costa, Debora Russo, Serena Vella, Claudio Russo, Alessandro Poggi, Antonella Brizzolara, Ilaria Penna, Andrea Mescola, Paola Menichini, Tullio Florio, Antonio Daga, Claudio Canale, Aldo Pagano, Arianna Gigoni, Federico Villa, and Mario Nizzari
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0301 basic medicine ,Pathology ,medicine.medical_specialty ,RNA, Untranslated ,Time Factors ,Cytoskeleton organization ,DNA Repair ,non-coding RNA ,Down-Regulation ,Mice, SCID ,Biology ,medicine.disease_cause ,Transfection ,Metastasis ,03 medical and health sciences ,neuroblastoma ,Downregulation and upregulation ,Cell Movement ,Mice, Inbred NOD ,Neuroblastoma ,Cell Line, Tumor ,medicine ,Cell Adhesion ,GTSE1 ,Animals ,Humans ,metastasis ,Neoplasm Invasiveness ,RNA, Antisense ,Cell adhesion ,Non-coding RNA ,Tumorigenesis ,Oncology ,Cell Proliferation ,Cell growth ,medicine.disease ,Tumor Burden ,Gene Expression Regulation, Neoplastic ,tumorigenesis ,030104 developmental biology ,HEK293 Cells ,Ki-67 Antigen ,Cancer research ,Carcinogenesis ,Microtubule-Associated Proteins ,DNA Damage ,Signal Transduction ,Research Paper - Abstract
// Ilaria Penna 1, 3, * , Arianna Gigoni 1, 3, * , Delfina Costa 1, 3, * , Serena Vella 4, * , Debora Russo 3 , Alessandro Poggi 3 , Federico Villa 1, 3 , Antonella Brizzolara 1, 3 , Claudio Canale 5 , Andrea Mescola 5 , Antonio Daga 3 , Claudio Russo 6 , Mario Nizzari 2 , Tullio Florio 2, 7 , Paola Menichini 3 , Aldo Pagano 1, 3 1 Department of Experimental Medicine (DIMES), University of Genova, Genova, Italy 2 Department of Internal Medicine (DIMI), University of Genova, Genova, Italy 3 IRCCS-AOU San Martino-IST, Genova, Italy 4 Department of Laboratory Medicine and Advanced Biotechnologies, IRCCS-ISMETT (Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione), Palermo, Italy 5 Nanophysics Unit, Italian Institute of Technology, Morego, Genova, Italy 6 Department of Health Sciences, University of Molise, Campobasso, Italy 7 Center of Excellence for Biomedical Research (CEBR), University of Genova, Genova, Italy * These authors have contributed equally to this work Correspondence to: Aldo Pagano, email: aldo.pagano@unige.it Keywords: neuroblastoma, non-coding RNA, GTSE1, tumorigenesis, metastasis Received: March 04, 2016 Accepted: November 23, 2016 Published: December 24, 2016 ABSTRACT We recently reported the in vitro over-expression of 45A, a RNA polymerase III-transcribed non-coding (nc)RNA, that perturbs the intracellular content of FE65L1 affecting cell proliferation rate, short-term response to genotoxic stress, substrate adhesion capacity and, ultimately, increasing the tumorigenic potential of human neuroblastoma cells. In this work, to deeply explore the mechanism by which 45A ncRNA contributes to cancer development, we targeted in vitro and in vivo 45A levels by the stable overexpression of antisense 45A RNA. 45A downregulation leads to deep modifications of cytoskeleton organization, adhesion and migration of neuroblastoma cells. These effects are correlated with alterations in the expression of several genes including GTSE1 (G2 and S phase-expressed-1), a crucial regulator of tumor cell migration and metastatic potential. Interestingly, the downregulation of 45A ncRNA strongly affects the in vivo tumorigenic potential of SKNBE2 neuroblastoma cells, increasing tumor nodule compactness and reducing GTSE1 protein expression in a subcutaneous neuroblastoma mouse model. Moreover, intracardiac injection of neuroblastoma cells showed that downregulation of 45A ncRNA also influences tumor metastatic ability. In conclusion, our data highlight a key role of 45A ncRNA in cancer development and suggest that its modulation might represent a possible novel anticancer therapeutic approach.
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