42 results on '"Elisa Dalla Pozza"'
Search Results
2. Mitochondrial Features of Mouse Myoblasts Are Finely Tuned by Low Doses of Ozone: The Evidence In Vitro
- Author
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Chiara Rita Inguscio, Elisa Dalla Pozza, Ilaria Dando, Federico Boschi, Gabriele Tabaracci, Osvaldo Angelini, Pietro Maria Picotti, Manuela Malatesta, and Barbara Cisterna
- Subjects
medical ozone ,reactive oxygen species ,mitochondrial cristae ,nuclear factor erythroid 2-related factor 2 (Nrf2) ,fluorescence microscopy ,transmission electron microscopy ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
The mild oxidative stress induced by low doses of gaseous ozone (O3) activates the antioxidant cell response through the nuclear factor erythroid 2-related factor 2 (Nrf2), thus inducing beneficial effects without cell damage. Mitochondria are sensitive to mild oxidative stress and represent a susceptible O3 target. In this in vitro study, we investigated the mitochondrial response to low O3 doses in the immortalized, non-tumoral muscle C2C12 cells; a multimodal approach including fluorescence microscopy, transmission electron microscopy and biochemistry was used. Results demonstrated that mitochondrial features are finely tuned by low O3 doses. The O3 concentration of 10 μg maintained normal levels of mitochondria-associated Nrf2, promoted the mitochondrial increase of size and cristae extension, reduced cellular reactive oxygen species (ROS) and prevented cell death. Conversely, in 20 μg O3-treated cells, where the association of Nrf2 with the mitochondria drastically dropped, mitochondria underwent more significant swelling, and ROS and cell death increased. This study, therefore, adds original evidence for the involvement of Nrf2 in the dose-dependent response to low O3 concentrations not only as an Antioxidant Response Elements (ARE) gene activator but also as a regulatory/protective factor of mitochondrial function.
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- 2023
- Full Text
- View/download PDF
3. Mitochondrial Dynamics as Potential Modulators of Hormonal Therapy Effectiveness in Males
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Andrea Errico, Sara Vinco, Giulia Ambrosini, Elisa Dalla Pozza, Nunzio Marroncelli, Nicola Zampieri, and Ilaria Dando
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andrology ,hormonal therapy ,mitochondrial dynamics ,male infertility ,Biology (General) ,QH301-705.5 - Abstract
Worldwide the incidence of andrological diseases is rising every year and, together with it, also the interest in them is increasing due to their strict association with disorders of the reproductive system, including impairment of male fertility, alterations of male hormones production, and/or sexual function. Prevention and early diagnosis of andrological dysfunctions have long been neglected, with the consequent increase in the incidence and prevalence of diseases otherwise easy to prevent and treat if diagnosed early. In this review, we report the latest evidence of the effect of andrological alterations on fertility potential in both young and adult patients, with a focus on the link between gonadotropins’ mechanism of action and mitochondria. Indeed, mitochondria are highly dynamic cellular organelles that undergo rapid morphological adaptations, conditioning a multitude of aspects, including their size, shape, number, transport, cellular distribution, and, consequently, their function. Since the first step of steroidogenesis takes place in these organelles, we consider that mitochondria dynamics might have a possible role in a plethora of signaling cascades, including testosterone production. In addition, we also hypothesize a central role of mitochondria fission boost on the decreased response to the commonly administrated hormonal therapy used to treat urological disease in pediatric and adolescent patients as well as infertile adults.
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- 2023
- Full Text
- View/download PDF
4. Integrated lipidomics and proteomics reveal cardiolipin alterations, upregulation of HADHA and long chain fatty acids in pancreatic cancer stem cells
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Claudia Di Carlo, Bebiana C. Sousa, Marcello Manfredi, Jessica Brandi, Elisa Dalla Pozza, Emilio Marengo, Marta Palmieri, Ilaria Dando, Michael J. O. Wakelam, Andrea F. Lopez-Clavijo, and Daniela Cecconi
- Subjects
Medicine ,Science - Abstract
Abstract Pancreatic cancer stem cells (PCSCs) play a key role in the aggressiveness of pancreatic ductal adenocarcinomas (PDAC); however, little is known about their signaling and metabolic pathways. Here we show that PCSCs have specific and common proteome and lipidome modulations. PCSCs displayed downregulation of lactate dehydrogenase A chain, and upregulation of trifunctional enzyme subunit alpha. The upregulated proteins of PCSCs are mainly involved in fatty acid (FA) elongation and biosynthesis of unsaturated FAs. Accordingly, lipidomics reveals an increase in long and very long-chain unsaturated FAs, which are products of fatty acid elongase-5 predicted as a key gene. Moreover, lipidomics showed the induction in PCSCs of molecular species of cardiolipin with mixed incorporation of 16:0, 18:1, and 18:2 acyl chains. Our data indicate a crucial role of FA elongation and alteration in cardiolipin acyl chain composition in PCSCs, representing attractive therapeutic targets in PDAC.
- Published
- 2021
- Full Text
- View/download PDF
5. Mitochondrial Elongation and OPA1 Play Crucial Roles during the Stemness Acquisition Process in Pancreatic Ductal Adenocarcinoma
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Cristian Andres Carmona-Carmona, Elisa Dalla Pozza, Giulia Ambrosini, Barbara Cisterna, Marta Palmieri, Ilaria Decimo, José M. Cuezva, Emanuela Bottani, and Ilaria Dando
- Subjects
cancer stem cells ,pancreatic ductal adenocarcinoma ,mitochondrial dynamics ,mitochondrial fusion ,OPA1 ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is the most common type of pancreatic cancer with an overall 5-year survival rate of less than 9%. The high aggressiveness of PDAC is linked to the presence of a subpopulation of cancer cells with a greater tumorigenic capacity, generically called cancer stem cells (CSCs). CSCs present a heterogeneous metabolic profile that might be supported by an adaptation of mitochondrial function; however, the role of this organelle in the development and maintenance of CSCs remains controversial. To determine the role of mitochondria in CSCs over longer periods, which may reflect more accurately their quiescent state, we studied the mitochondrial physiology in CSCs at short-, medium-, and long-term culture periods. We found that CSCs show a significant increase in mitochondrial mass, more mitochondrial fusion, and higher mRNA expression of genes involved in mitochondrial biogenesis than parental cells. These changes are accompanied by a regulation of the activities of OXPHOS complexes II and IV. Furthermore, the protein OPA1, which is involved in mitochondrial dynamics, is overexpressed in CSCs and modulates the tumorsphere formation. Our findings indicate that CSCs undergo mitochondrial remodeling during the stemness acquisition process, which could be exploited as a promising therapeutic target against pancreatic CSCs.
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- 2022
- Full Text
- View/download PDF
6. 3-Bromo-Isoxazoline Derivatives Inhibit GAPDH Enzyme in PDAC Cells Triggering Autophagy and Apoptotic Cell Death
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Raffaella Pacchiana, Nidula Mullappilly, Andrea Pinto, Stefania Bova, Stefania Forciniti, Gregorio Cullia, Elisa Dalla Pozza, Emanuela Bottani, Ilaria Decimo, Ilaria Dando, Stefano Bruno, Paola Conti, and Massimo Donadelli
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pancreas cancer ,cancer metabolism ,GAPDH ,apoptosis ,autophagy ,cell death ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
A growing interest in the study of aerobic glycolysis as a key pathway for cancer-cell energetic metabolism, favouring tumour progression and invasion, has led to consider GAPDH as an effective drug target to specifically hit cancer cells. In this study, we have investigated a panel of 3-bromo-isoxazoline derivatives based on previously identified inhibitors of Plasmodium falciparum GAPDH (PfGAPDH). The compounds are active, to a different extent, as inhibitors of human-recombinant GAPDH. They showed an antiproliferative effect on pancreatic ductal-adenocarcinoma cells (PDAC) and pancreatic-cancer stem cells (CSCs), and among them two promising compounds were selected to be tested in vivo. Interestingly, these compounds were not effective in fibroblasts. The AXP-3019 derivative was able to block PDAC-cell growth in mice xenograft without apparent toxicity. The overall results support the assumption that selective inhibition of the glycolytic pathway, by targeting GAPDH, is an effective therapy for pancreatic cancer and that 3-bromo-isoxazoline derivatives represent a new class of anti-cancer compounds targeting glycolysis.
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- 2022
- Full Text
- View/download PDF
7. Divergent Roles of Mitochondria Dynamics in Pancreatic Ductal Adenocarcinoma
- Author
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Cristian Andres Carmona-Carmona, Elisa Dalla Pozza, Giulia Ambrosini, Andrea Errico, and Ilaria Dando
- Subjects
mitochondrial dynamics ,PDAC ,cancer stem cells ,metabolism ,molecular target ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive tumors; it is often diagnosed at an advanced stage and is hardly treatable. These issues are strictly linked to the absence of early diagnostic markers and the low efficacy of treatment approaches. Recently, the study of the metabolic alterations in cancer cells has opened the way to important findings that can be exploited to generate new potential therapies. Within this scenario, mitochondria represent important organelles within which many essential functions are necessary for cell survival, including some key reactions involved in energy metabolism. These organelles remodel their shape by dividing or fusing themselves in response to cellular needs or stimuli. Interestingly, many authors have shown that mitochondrial dynamic equilibrium is altered in many different tumor types. However, up to now, it is not clear whether PDAC cells preferentially take advantage of fusion or fission processes since some studies reported a wide range of different results. This review described the role of both mitochondria arrangement processes, i.e., fusion and fission events, in PDAC, showing that a preference for mitochondria fragmentation could sustain tumor needs. In addition, we also highlight the importance of considering the metabolic arrangement and mitochondria assessment of cancer stem cells, which represent the most aggressive tumor cell type that has been shown to have distinctive metabolic features to that of differentiated tumor cells.
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- 2022
- Full Text
- View/download PDF
8. Extracellular Matrix Composition Modulates the Responsiveness of Differentiated and Stem Pancreatic Cancer Cells to Lipophilic Derivate of Gemcitabine
- Author
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Stefania Forciniti, Elisa Dalla Pozza, Maria Raffaella Greco, Tiago Miguel Amaral Carvalho, Barbara Rolando, Giulia Ambrosini, Cristian Andres Carmona-Carmona, Raffaella Pacchiana, Daria Di Molfetta, Massimo Donadelli, Silvia Arpicco, Marta Palmieri, Stephan Joel Reshkin, Ilaria Dando, and Rosa Angela Cardone
- Subjects
pancreatic ductal adenocarcinoma ,cancer stem cells ,3D organotypic cultures ,gemcitabine ,prodrug ,extracellular matrix ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Background: Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal disease. Gemcitabine (GEM) is used as the gold standard drug in PDAC treatment. However, due to its poor efficacy, it remains urgent to identify novel strategies to overcome resistance issues. In this context, an intense stroma reaction and the presence of cancer stem cells (CSCs) have been shown to influence PDAC aggressiveness, metastatic potential, and chemoresistance. Methods: We used three-dimensional (3D) organotypic cultures grown on an extracellular matrix composed of Matrigel or collagen I to test the effect of the new potential therapeutic prodrug 4-(N)-stearoyl-GEM, called C18GEM. We analyzed C18GEM cytotoxic activity, intracellular uptake, apoptosis, necrosis, and autophagy induction in both Panc1 cell line (P) and their derived CSCs. Results: PDAC CSCs show higher sensitivity to C18GEM treatment when cultured in both two-dimensional (2D) and 3D conditions, especially on collagen I, in comparison to GEM. The intracellular uptake mechanisms of C18GEM are mainly due to membrane nucleoside transporters’ expression and fatty acid translocase CD36 in Panc1 P cells and to clathrin-mediated endocytosis and CD36 in Panc1 CSCs. Furthermore, C18GEM induces an increase in cell death compared to GEM in both cell lines grown on 2D and 3D cultures. Finally, C18GEM stimulated protective autophagy in Panc1 P and CSCs cultured on 3D conditions. Conclusion: We propose C18GEM together with autophagy inhibitors as a valid alternative therapeutic approach in PDAC treatment.
- Published
- 2020
- Full Text
- View/download PDF
9. Mutant p53 proteins counteract autophagic mechanism sensitizing cancer cells to mTOR inhibition
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Marco Cordani, Elisa Oppici, Ilaria Dando, Elena Butturini, Elisa Dalla Pozza, Mercedes Nadal-Serrano, Jordi Oliver, Pilar Roca, Sofia Mariotto, Barbara Cellini, Giovanni Blandino, Marta Palmieri, Silvia Di Agostino, and Massimo Donadelli
- Subjects
Autophagy ,Mutant p53 ,Gain‐of‐function ,Cancer ,mTOR ,AMPK ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Mutations in TP53 gene play a pivotal role in tumorigenesis and cancer development. Here, we report that gain‐of‐function mutant p53 proteins inhibit the autophagic pathway favoring antiapoptotic effects as well as proliferation of pancreas and breast cancer cells. We found that mutant p53 significantly counteracts the formation of autophagic vesicles and their fusion with lysosomes throughout the repression of some key autophagy‐related proteins and enzymes as BECN1 (and P‐BECN1), DRAM1, ATG12, SESN1/2 and P‐AMPK with the concomitant stimulation of mTOR signaling. As a paradigm of this mechanism, we show that atg12 gene repression was mediated by the recruitment of the p50 NF‐κB/mutant p53 protein complex onto the atg12 promoter. Either mutant p53 or p50 NF‐κB depletion downregulates atg12 gene expression. We further correlated the low expression levels of autophagic genes (atg12, becn1, sesn1, and dram1) with a reduced relapse free survival (RFS) and distant metastasis free survival (DMFS) of breast cancer patients carrying TP53 gene mutations conferring a prognostic value to this mutant p53‐and autophagy‐related signature. Interestingly, the mutant p53‐driven mTOR stimulation sensitized cancer cells to the treatment with the mTOR inhibitor everolimus. All these results reveal a novel mechanism through which mutant p53 proteins promote cancer cell proliferation with the concomitant inhibition of autophagy.
- Published
- 2016
- Full Text
- View/download PDF
10. Progressively De-Differentiated Pancreatic Cancer Cells Shift from Glycolysis to Oxidative Metabolism and Gain a Quiescent Stem State
- Author
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Giulia Ambrosini, Elisa Dalla Pozza, Giuseppina Fanelli, Claudia Di Carlo, Andrea Vettori, Giuseppe Cannino, Chiara Cavallini, Cristian Andres Carmona-Carmona, Jessica Brandi, Sara Rinalducci, Maria Teresa Scupoli, Andrea Rasola, Daniela Cecconi, Marta Palmieri, and Ilaria Dando
- Subjects
cancer stem cells ,pancreatic ductal adenocarcinoma ,cancer metabolism ,quiescence ,metabolic plasticity ,Cytology ,QH573-671 - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is typically characterized by high chemoresistance and metastatic spread, features mainly attributable to cancer stem cells (CSCs). It is of central interest the characterization of CSCs and, in particular, the study of their metabolic features in order to selectively identify their peculiarities for an efficient therapeutic approach. In this study, CSCs have been obtained by culturing different PDAC cell lines with a specific growth medium. Cells were characterized for the typical stem/mesenchymal properties at short-, medium-, and long-term culture. Metabolomics, proteomics, analysis of oxygen consumption rate in live cells, and the effect of the inhibition of lactate transporter on cell proliferation have been performed to delineate the metabolism of CSCs. We show that gradually de-differentiated pancreatic cancer cells progressively increase the expression of both stem and epithelial-to-mesenchymal transition markers, shift their metabolism from a glycolytic to an oxidative one, and lastly gain a quiescent state. These quiescent stem cells are characterized by high chemo-resistance, clonogenic ability, and metastatic potential. Re-differentiation reverts these features, re-activating their proliferative capacity and glycolytic metabolism, which generally correlates with high aggressiveness. These observations add an important piece of knowledge to the comprehension of the biology of CSCs, whose metabolic plasticity could be exploited for the generation of promising and selective therapeutic approaches for PDAC patients.
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- 2020
- Full Text
- View/download PDF
11. Genetic and epigenetic mechanisms regulating catalase expression in chronic lymphocytic leukemia
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Marilisa Galasso, Elisa Dalla Pozza, Roberto Chignola, Simona Gambino, Chiara Cavallini, Francesca Maria Quaglia, Ornella Lovato, Ilaria Dando, Giorgio Malpeli, Mauro Krampera, Massimo Donadelli, Maria Grazia Romanelli, and Maria Teresa Scupoli
- Abstract
Chronic lymphocytic leukemia (CLL) is an incurable disease characterized by an extremely variable clinical course. We have recently shown that high catalase (CAT) expression identifies patients with an aggressive clinical course. Elucidating mechanisms regulating CAT expression in CLL is preeminent to understand disease mechanisms and develop strategies for improving its clinical management. In this study, we investigated the role of the CAT promoter rs1001179 single nucleotide polymorphism (SNP) and of the CpG Island II methylation encompassing this SNP in the regulation of CAT expression in CLL. Leukemic cells harboring the rs1001179 SNP T allele exhibited a significantly higher CAT expression compared with cells bearing the CC genotype. CAT promoter harboring the T -but not C- allele was accessible to ETS-1 and GR-β transcription factors. Moreover, CLL cells exhibited lower methylation levels than normal B cells, in line with the higher CAT mRNA and protein expressed by CLL in comparison with normal B cells. Methylation levels at specific CpG sites negatively correlated with CAT levels in CLL cells. Inhibition of methyltransferase activity induced a significant increase of CAT levels, thus functionally validating the role of CpG methylation in regulating CAT expression in CLL. Finally, the CT/TT genotypes were associated with lower methylation and higher CAT levels, suggesting that the rs1001179 T allele and CpG methylation may interact in regulating CAT expression in CLL. This study identifies genetic and epigenetic mechanisms underlying differential expression of CAT, which could be of crucial relevance for the development of therapies targeting redox regulatory pathways in CLL.
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- 2022
12. The rs1001179 SNP and CpG methylation regulate catalase expression in chronic lymphocytic leukemia
- Author
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Marilisa Galasso, Elisa Dalla Pozza, Roberto Chignola, Simona Gambino, Chiara Cavallini, Francesca Maria Quaglia, Ornella Lovato, Ilaria Dando, Giorgio Malpeli, Mauro Krampera, Massimo Donadelli, Maria G. Romanelli, and Maria T. Scupoli
- Subjects
Pharmacology ,DNA methylation ,Methyltransferases ,Cell Biology ,Catalase ,Leukemia, Lymphocytic, Chronic, B-Cell ,Polymorphism, Single Nucleotide ,Single nucleotide polymorphism ,Cellular and Molecular Neuroscience ,Humans ,Molecular Medicine ,Chronic lymphocytic leukemia ,RNA, Messenger ,Molecular Biology ,Transcription Factors - Abstract
Chronic lymphocytic leukemia (CLL) is an incurable disease characterized by an extremely variable clinical course. We have recently shown that high catalase (CAT) expression identifies patients with an aggressive clinical course. Elucidating mechanisms regulating CAT expression in CLL is preeminent to understand disease mechanisms and develop strategies for improving its clinical management. In this study, we investigated the role of the CAT promoter rs1001179 single nucleotide polymorphism (SNP) and of the CpG Island II methylation encompassing this SNP in the regulation of CAT expression in CLL. Leukemic cells harboring the rs1001179 SNP T allele exhibited a significantly higher CAT expression compared with cells bearing the CC genotype. CAT promoter harboring the T -but not C- allele was accessible to ETS-1 and GR-β transcription factors. Moreover, CLL cells exhibited lower methylation levels than normal B cells, in line with the higher CAT mRNA and protein expressed by CLL in comparison with normal B cells. Methylation levels at specific CpG sites negatively correlated with CAT levels in CLL cells. Inhibition of methyltransferase activity induced a significant increase in CAT levels, thus functionally validating the role of CpG methylation in regulating CAT expression in CLL. Finally, the CT/TT genotypes were associated with lower methylation and higher CAT levels, suggesting that the rs1001179 T allele and CpG methylation may interact in regulating CAT expression in CLL. This study identifies genetic and epigenetic mechanisms underlying differential expression of CAT, which could be of crucial relevance for the development of therapies targeting redox regulatory pathways in CLL. Graphical abstract
- Published
- 2022
13. Extracellular Matrix Composition Modulates the Responsiveness of Differentiated and Stem Pancreatic Cancer Cells to Lipophilic Derivate of Gemcitabine
- Author
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Elisa Dalla Pozza, Massimo Donadelli, Giulia Ambrosini, Cristian Andres Carmona-Carmona, Daria Di Molfetta, Ilaria Dando, Stefania Forciniti, Raffaella Pacchiana, Stephan J. Reshkin, Maria Raffaella Greco, Marta Palmieri, Rosa Angela Cardone, Tiago M.A. Carvalho, Barbara Rolando, and Silvia Arpicco
- Subjects
0301 basic medicine ,cancer stem cells ,endocrine system diseases ,Apoptosis ,Deoxycytidine ,lcsh:Chemistry ,0302 clinical medicine ,Cytotoxic T cell ,Prodrugs ,lcsh:QH301-705.5 ,Spectroscopy ,Chemistry ,gemcitabine ,chemoresistance ,Cell Differentiation ,General Medicine ,3D organotypic cultures ,extracellular matrix ,pancreatic ductal adenocarcinoma ,prodrug ,3. Good health ,Computer Science Applications ,Drug Combinations ,030220 oncology & carcinogenesis ,Neoplastic Stem Cells ,Proteoglycans ,Collagen ,Intracellular ,Carcinoma, Pancreatic Ductal ,Catalysis ,Collagen Type I ,Article ,Inorganic Chemistry ,03 medical and health sciences ,Organ Culture Techniques ,Cancer stem cell ,Pancreatic cancer ,Cell Line, Tumor ,medicine ,Autophagy ,Humans ,Physical and Theoretical Chemistry ,Molecular Biology ,Matrigel ,Organic Chemistry ,medicine.disease ,Pancreatic Neoplasms ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,Cell culture ,Drug Resistance, Neoplasm ,Cancer research ,Laminin - Abstract
Background: Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal disease. Gemcitabine (GEM) is used as the gold standard drug in PDAC treatment. However, due to its poor efficacy, it remains urgent to identify novel strategies to overcome resistance issues. In this context, an intense stroma reaction and the presence of cancer stem cells (CSCs) have been shown to influence PDAC aggressiveness, metastatic potential, and chemoresistance. Methods: We used three-dimensional (3D) organotypic cultures grown on an extracellular matrix composed of Matrigel or collagen I to test the effect of the new potential therapeutic prodrug 4-(N)-stearoyl-GEM, called C18GEM. We analyzed C18GEM cytotoxic activity, intracellular uptake, apoptosis, necrosis, and autophagy induction in both Panc1 cell line (P) and their derived CSCs. Results: PDAC CSCs show higher sensitivity to C18GEM treatment when cultured in both two-dimensional (2D) and 3D conditions, especially on collagen I, in comparison to GEM. The intracellular uptake mechanisms of C18GEM are mainly due to membrane nucleoside transporters&rsquo, expression and fatty acid translocase CD36 in Panc1 P cells and to clathrin-mediated endocytosis and CD36 in Panc1 CSCs. Furthermore, C18GEM induces an increase in cell death compared to GEM in both cell lines grown on 2D and 3D cultures. Finally, C18GEM stimulated protective autophagy in Panc1 P and CSCs cultured on 3D conditions. Conclusion: We propose C18GEM together with autophagy inhibitors as a valid alternative therapeutic approach in PDAC treatment.
- Published
- 2021
14. Promising 3D in vitro models for studying tumour heterogeneity and testing novel therapeutic approaches in pancreatic cancer
- Author
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Elisa Dalla Pozza, Marta Cavo, Marta Palmieri, Stefania Forciniti, Loretta L. del Mercato, and Ilaria Dando
- Subjects
Pancreatic ductal adenocarcinoma ,Tumour heterogeneity ,Cancer stem cell ,business.industry ,Pancreatic cancer ,embryonic structures ,medicine ,Cancer research ,General Medicine ,medicine.disease ,business ,In vitro - Abstract
In this study we produced 3D organotypic cultures and spheroids to mimic the complex microenvironment of pancreatic cancer and to test alternative therapeutic strategies.
- Published
- 2021
15. Effects of CD20 antibodies and kinase inhibitors on B-cell receptor signalling and survival of chronic lymphocytic leukaemia cells
- Author
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Chiara Cavallini, Massimo Donadelli, Giovanni Pizzolo, Maria Teresa Scupoli, Mauro Krampera, Maria Grazia Romanelli, Ornella Lovato, Marilisa Galasso, Ilaria Dando, Elisa Dalla Pozza, and Roberto Chignola
- Subjects
B-cell receptor ,Receptors, Antigen, B-Cell ,Apoptosis ,Ofatumumab ,Antibodies, Monoclonal, Humanized ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Antineoplastic Agents, Immunological ,Piperidines ,immune system diseases ,Obinutuzumab ,hemic and lymphatic diseases ,Antineoplastic Combined Chemotherapy Protocols ,Medicine ,Humans ,Protein Kinase Inhibitors ,Quinazolinones ,CD20 ,biology ,business.industry ,Adenine ,breakpoint cluster region ,Hematology ,Antigens, CD20 ,Leukemia, Lymphocytic, Chronic, B-Cell ,chemistry ,Purines ,030220 oncology & carcinogenesis ,Ibrutinib ,leukaemia ,biology.protein ,Cancer research ,CD20 mAb ,Rituximab ,business ,Idelalisib ,chronic lymphocytic leukaemia ,signal transduction ,030215 immunology ,medicine.drug - Abstract
Recently, clinical trial results have established inhibitors of B-cell receptor (BCR)-associated kinase (BAKi), with or without CD20 moniclonal antibodies (mAbs), as the preferred first-line treatment for most chronic lymphocytic leukaemia (CLL) patients. Using phosphospecific flow cytometry, we showed that in leukaemic cells from CLL patients the CD20 therapeutic antibodies - rituximab, ofatumumab, and obinutuzumab - inhibited BCR signalling pathways targeting preferentially pBTKY551 - but not BTKY223 - and pAKT. On the contrary, ibrutinib and idelalisib reduced pBTKY223 to a higher extent than pBTKY551 . The strong reduction of pAKT induced by idelalisib was enhanced by its combination with rituximab or ofatumumab. Moreover, CD20 mAbs and BAKi induced the death of leukaemia cells that was significantly potentiated by their combination. Analysis of the enhancement of cell death in these combinations revealed an approximately additive enhancement induced by rituximab or obinutuzumab combined with ibrutinib or idelalisib. Taken together, our data identified negative regulatory effects of CD20 mAbs and their combinations with BAKi on BCR signalling and cell survival in CLL. In conclusion, this study advances our understanding of mechanisms of action of CD20 mAbs as single agents or in combination with BAKi and could inform on the potential of combined therapies in ongoing and future clinical trials in patients with CLL.
- Published
- 2021
16. Integrated lipidomics and proteomics reveal cardiolipin alterations, upregulation of HADHA and long chain fatty acids in pancreatic cancer stem cells
- Author
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Marta Palmieri, Jessica Brandi, Bebiana C. Sousa, Emilio Marengo, Elisa Dalla Pozza, Michael J.O. Wakelam, Andrea F. Lopez-Clavijo, Ilaria Dando, Marcello Manfredi, Claudia Di Carlo, Daniela Cecconi, Cecconi, Daniela [0000-0002-7314-8941], and Apollo - University of Cambridge Repository
- Subjects
Cardiolipins ,Science ,Lactate dehydrogenase A ,pancreatic cancer stem cells (PCSCs) ,Proteomics ,therapeutic targets ,Article ,chemistry.chemical_compound ,proteomics ,Downregulation and upregulation ,Lipidomics ,Cardiolipin ,Humans ,education ,chemistry.chemical_classification ,education.field_of_study ,Multidisciplinary ,Cancer stem cells ,fatty acid elongase-5 ,pancreatic ductal adenocarcinomas ,Fatty acid ,Lipidome ,Lipid Metabolism ,Cancer metabolism ,Up-Regulation ,Pancreatic Neoplasms ,chemistry ,Biochemistry ,Proteome ,Neoplastic Stem Cells ,Medicine ,lipidomics ,Mitochondrial Trifunctional Protein, alpha Subunit ,cardiolipin ,Carcinoma, Pancreatic Ductal - Abstract
Pancreatic cancer stem cells (PCSCs) play a key role in the aggressiveness of pancreatic ductal adenocarcinomas (PDAC); however, little is known about their signaling and metabolic pathways. Here we show that PCSCs have specific and common proteome and lipidome modulations. PCSCs displayed downregulation of lactate dehydrogenase A chain, and upregulation of trifunctional enzyme subunit alpha. The upregulated proteins of PCSCs are mainly involved in fatty acid (FA) elongation and biosynthesis of unsaturated FAs. Accordingly, lipidomics reveals an increase in long and very long-chain unsaturated FAs, which are products of fatty acid elongase-5 predicted as a key gene. Moreover, lipidomics showed the induction in PCSCs of molecular species of cardiolipin with mixed incorporation of 16:0, 18:1, and 18:2 acyl chains. Our data indicate a crucial role of FA elongation and alteration in cardiolipin acyl chain composition in PCSCs, representing attractive therapeutic targets in PDAC.
- Published
- 2021
17. Progressively De-Differentiated Pancreatic Cancer Cells Shift from Glycolysis to Oxidative Metabolism and Gain a Quiescent Stem State
- Author
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Jessica Brandi, Cristian Andres Carmona-Carmona, Chiara Cavallini, Andrea Vettori, Ilaria Dando, Giuseppe Cannino, Sara Rinalducci, Daniela Cecconi, Marta Palmieri, Maria Teresa Scupoli, Andrea Rasola, Claudia Di Carlo, Giuseppina Fanelli, Elisa Dalla Pozza, and Giulia Ambrosini
- Subjects
cancer stem cells ,cancer metabolism ,pancreatic ductal adenocarcinoma ,Biology ,Proteomics ,Article ,Oxygen Consumption ,Cancer stem cell ,Cell Line, Tumor ,Pancreatic cancer ,medicine ,Animals ,Humans ,quiescence ,Clonogenic assay ,lcsh:QH301-705.5 ,Cellular Senescence ,Zebrafish ,Cell Proliferation ,Cell growth ,Cell Cycle ,Mesenchymal stem cell ,Cell Differentiation ,General Medicine ,medicine.disease ,metabolic plasticity ,Pancreatic Neoplasms ,lcsh:Biology (General) ,Cell culture ,Neoplastic Stem Cells ,Cancer research ,Stem cell ,Glycolysis ,Carcinoma, Pancreatic Ductal - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is typically characterized by high chemoresistance and metastatic spread, features mainly attributable to cancer stem cells (CSCs). It is of central interest the characterization of CSCs and, in particular, the study of their metabolic features in order to selectively identify their peculiarities for an efficient therapeutic approach. In this study, CSCs have been obtained by culturing different PDAC cell lines with a specific growth medium. Cells were characterized for the typical stem/mesenchymal properties at short-, medium-, and long-term culture. Metabolomics, proteomics, analysis of oxygen consumption rate in live cells, and the effect of the inhibition of lactate transporter on cell proliferation have been performed to delineate the metabolism of CSCs. We show that gradually de-differentiated pancreatic cancer cells progressively increase the expression of both stem and epithelial-to-mesenchymal transition markers, shift their metabolism from a glycolytic to an oxidative one, and lastly gain a quiescent state. These quiescent stem cells are characterized by high chemo-resistance, clonogenic ability, and metastatic potential. Re-differentiation reverts these features, re-activating their proliferative capacity and glycolytic metabolism, which generally correlates with high aggressiveness. These observations add an important piece of knowledge to the comprehension of the biology of CSCs, whose metabolic plasticity could be exploited for the generation of promising and selective therapeutic approaches for PDAC patients.
- Published
- 2020
18. Regulation of succinate dehydrogenase and role of succinate in cancer
- Author
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Massimo Donadelli, Marta Palmieri, Elio Liboi, Raffaella Pacchiana, Maria Teresa Scupoli, Ilaria Dando, and Elisa Dalla Pozza
- Subjects
0301 basic medicine ,Epithelial-Mesenchymal Transition ,Angiogenesis ,Succinic Acid ,macromolecular substances ,SDH ,Biology ,medicine.disease_cause ,03 medical and health sciences ,0302 clinical medicine ,Neoplasms ,medicine ,cancer ,Animals ,Humans ,Epithelial–mesenchymal transition ,Epigenetics ,Succinate dehydrogenase ,Cell migration ,Cell Biology ,Metabolism ,succinate ,oncometabolites ,Citric acid cycle ,Succinate Dehydrogenase ,030104 developmental biology ,Biochemistry ,metabolism ,biology.protein ,Carcinogenesis ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Succinate dehydrogenase (SDH) has been classically considered a mitochondrial enzyme with the unique property to participate in both the citric acid cycle and the electron transport chain. However, in recent years, several studies have highlighted the role of the SDH substrate, i.e. succinate, in biological processes other than metabolism, tumorigenesis being the most remarkable. For this reason, SDH has now been defined a tumor suppressor and succinate an oncometabolite. In this review, we discuss recent findings regarding alterations in SDH activity leading to succinate accumulation, which include SDH mutations, regulation of mRNA expression, post-translational modifications and endogenous SDH inhibitors. Further, we report an extensive examination of the role of succinate in cancer development through the induction of epigenetic and metabolic alterations and the effects on epithelial to mesenchymal transition, cell migration and invasion, and angiogenesis. Finally, we have focused on succinate and SDH as diagnostic markers for cancers having altered SDH expression/activity.
- Published
- 2019
19. Pancreatic cancer stem cell proliferation is strongly inhibited by diethyldithiocarbamate-copper complex loaded into hyaluronic acid decorated liposomes
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Alessandro Marengo, Nicolas Tsapis, Marta Palmieri, Ilaria Dando, Elias Fattal, Christopher Heeschen, Giuseppina Fanelli, Barbara Stella, Najet Yagoubi, Silvia Arpicco, Stefania Forciniti, Elisa Dalla Pozza, Institut Galien Paris-Sud (IGPS), Université Paris-Sud - Paris 11 (UP11)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Groupe matériaux et santé, Université Paris-Sud - Paris 11 (UP11), Istituto di chimica biologica, and University of Verona (UNIVR)
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PDAC patient-derived cells ,Pancreatic cancer stem cells ,Cell ,Biochemistry ,0302 clinical medicine ,CD44 ,Phospholipids ,0303 health sciences ,Liposome ,education.field_of_study ,biology ,Calorimetry, Differential Scanning ,Chemistry ,Diethyldithiocarbamate/copper complex ,Hyaluronic acid ,Liposomes ,Biophysics ,Molecular Biology ,3. Good health ,medicine.anatomical_structure ,Hyaluronan Receptors ,030220 oncology & carcinogenesis ,Neoplastic Stem Cells ,Stem cell ,Ditiocarb ,Carcinoma, Pancreatic Ductal ,Population ,03 medical and health sciences ,Microscopy, Electron, Transmission ,Pancreatic cancer ,Cell Line, Tumor ,medicine ,Humans ,education ,Pancreas ,030304 developmental biology ,Cell Proliferation ,Cryoelectron Microscopy ,medicine.disease ,Acetylcysteine ,Pancreatic Neoplasms ,[SDV.SP.PG]Life Sciences [q-bio]/Pharmaceutical sciences/Galenic pharmacology ,Cell culture ,Cancer cell ,biology.protein ,Cancer research ,Drug Screening Assays, Antitumor ,Reactive Oxygen Species ,Copper - Abstract
International audience; Background: Pancreatic cancer stem cells (CSCs) are responsible for resistance to standard therapy, metastatic potential, and disease relapse following treatments. The current therapy for pancreatic ductal adenocarcinoma (PDAC) preferentially targets the more differentiated cancer cell population, leaving CSCs as a cell source for tumor mass formation and recurrence. For this reason, there is an urgent need to improve current therapies and develop novel CSC-targeted therapeutic approaches. Methods: Hyaluronic acid (HA) decorated liposomes, containing diethyldithiocarbamate-copper (Cu(DDC) 2), able to target the specific CSC marker CD44 receptor were prepared by ion gradient technique and fully characterized. Their antiproliferative effect was evaluated on pancreatic CSCs derived from PDAC cell lines or patients. To clarify the mechanism of action of Cu(DDC) 2 liposomes, ROS level neutralization assay in the presence of N-acetyl-L-cysteine was performed. Results: Liposomes showed high encapsulation efficiency and Cryo-TEM analysis revealed the presence of Cu (DDC) 2 crystals in the aqueous core of liposomes. In vitro test on pancreatic CSCs derived from PDAC cell lines or patients showed high ROS mediated anticancer activity of HA decorated liposomes. The sphere formation capability of CSCs obtained from patients was drastically reduced by liposomal formulations containing Cu (DDC) 2. Conclusions: The obtained results show that the encapsulation of Cu(DDC) 2 complex in HA decorated liposomes strongly increases its anti-proliferative activity on pancreatic CSCs. General significance: This paper describes for the first time the use of HA decorated liposomes containing Cu (DDC) 2 against pancreatic CSCs and opens the way to the development of nanomedicine based CSC-targeted therapeutic approaches.
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- 2019
20. Oncometabolites in cancer aggressiveness and tumour repopulation
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Ilaria Dando, Elisa Dalla Pozza, Raffaella Pacchiana, Giovanna Butera, Massimo Donadelli, Marta Palmieri, Nidula Mullappilly, Giulia Ambrosini, and Margalida Torrens-Mas
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0106 biological sciences ,Cancer relapse ,2-hydroxyglutarate ,010603 evolutionary biology ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,Fumarate Hydratase ,03 medical and health sciences ,Recurrence ,Neoplasms ,Tumor Microenvironment ,Medicine ,cancer ,Animals ,Humans ,030304 developmental biology ,0303 health sciences ,fumarate ,biology ,business.industry ,Succinate dehydrogenase ,Cancer ,medicine.disease ,succinate ,Phenotype ,Isocitrate Dehydrogenase ,oncometabolites ,Succinate Dehydrogenase ,Isocitrate dehydrogenase ,Fumarase ,Cancer cell ,biology.protein ,Cancer research ,Repopulation ,General Agricultural and Biological Sciences ,business ,metabolism - Abstract
Tumour repopulation is recognized as a crucial event in tumour relapse where therapy-sensitive dying cancer cells influence the tumour microenvironment to sustain therapy-resistant cancer cell growth. Recent studies highlight the role of the oncometabolites succinate, fumarate, and 2-hydroxyglutarate in the aggressiveness of cancer cells and in the worsening of the patient's clinical outcome. These oncometabolites can be produced and secreted by cancer and/or surrounding cells, modifying the tumour microenvironment and sustaining an invasive neoplastic phenotype. In this review, we report recent findings concerning the role in cancer development of succinate, fumarate, and 2-hydroxyglutarate and the regulation of their related enzymes succinate dehydrogenase, fumarate hydratase, and isocitrate dehydrogenase. We propose that oncometabolites are crucially involved in tumour repopulation. The study of the mechanisms underlying the relationship between oncometabolites and tumour repopulation is fundamental for identifying efficient anti-cancer therapeutic strategies and novel serum biomarkers in order to overcome cancer relapse.
- Published
- 2018
21. Extracellular Matrix composition modulates PDAC parenchymal and stem cell plasticity and behavior through the secretome
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Elisa Dalla Pozza, Katrine Zeeberg, Ilaria Dando, Stefania Forciniti, Maria Mastrodonato, Maria Raffaella Greco, Stefania Cannone, Stephan J. Reshkin, Giulia Biondani, Valeria Casavola, Marta Palmieri, and Rosa Angela Cardone
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0301 basic medicine ,Vascular Endothelial Growth Factor A ,Angiogenesis ,Population ,Cell Plasticity ,Cell Culture Techniques ,Biology ,Adenocarcinoma ,Biochemistry ,Collagen Type I ,Extracellular matrix ,03 medical and health sciences ,Cancer stem cell ,Cell Line, Tumor ,Tumor Microenvironment ,Humans ,Vasculogenic mimicry ,Neoplasm Invasiveness ,Pancreatic Adenocarcinoma ,Neoplasm Metastasis ,education ,Molecular Biology ,Parenchymal Tissue ,Cell Proliferation ,education.field_of_study ,Tumor microenvironment ,Matrigel ,Neovascularization, Pathologic ,Cancer stem cells ,Desmoplastic Reaction ,Cell Biology ,Vascular Endothelial Growth Factor Receptor-2 ,3D organotypic cultures ,VEFGR-2 ,Extracellular Matrix ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Cancer research ,Neoplastic Stem Cells ,Stem cell ,Carcinoma, Pancreatic Ductal - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers. Its aggressiveness is driven by an intense fibrotic desmoplastic reaction in which the increasingly collagen I-rich extracellular matrix (ECM) and several cell types, including cancer stem cells (CSCs), create a tumor-supportive environment. However, how ECM composition regulates CSC dynamics and their relationship with the principle parenchymal tumor population to promote early invasive growth is not yet characterized. For this, we utilized a platform of 3D organotypic cultures composed of laminin-rich Matrigel, representative of an early tumor, plus increasing concentrations of collagen I to simulate malignant stroma progression. As ECM collagen I increases, CSCs progress from a rapidly growing, vascular phenotype to a slower growing, avascular phase, while maintaining their endothelial-like gene signatures. This transition is supported autocrinically by the CSCs and paracrinically by the parenchymal cells via their ECM-dependent secretomes. Indeed, when growing on an early tumor ECM, the CSCs are dedicated toward the preparation of a vascular niche by (a) activating their growth program, (b) secreting high levels of proangiogenic factors which stimulate both angiogenesis and vasculogenic mimicry, and (c) overexpressing VEGFR-2, which is activated by VEGF secreted by both the CSC and parenchymal cells. On Matrigel, the more differentiated parenchymal tumor cell population had reduced growth but a high invasive capacity. This concerted high local invasion of parenchymal cells into the CSC-derived vascular network suggests that a symbiotic relationship between the parenchymal cells and the CSCs underlies the initiation and maintenance of early PDAC infiltration and metastasis.
- Published
- 2018
22. Secreted molecules inducing epithelial-to-mesenchymal transition in cancer development
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Marta Palmieri, Stefania Forciniti, Elisa Dalla Pozza, and Ilaria Dando
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0301 basic medicine ,Epithelial-Mesenchymal Transition ,Biology ,Exosomes ,03 medical and health sciences ,Paracrine signalling ,Fibrosis ,Neoplasms ,Paracrine Communication ,Tumor Microenvironment ,medicine ,Humans ,cancer ,Epithelial–mesenchymal transition ,epithelial-to-mesenchymal transition (EMT) ,secreted molecules ,Mesenchymal stem cell ,Cancer ,Mesenchymal Stem Cells ,Cell Biology ,medicine.disease ,Phenotype ,Microvesicles ,Cell biology ,Gene Expression Regulation, Neoplastic ,Autocrine Communication ,MicroRNAs ,030104 developmental biology ,Wound healing ,Developmental Biology - Abstract
The epithelial-mesenchymal transition (EMT) is a biologic process that allows a polarized epithelial cell to undergo multiple biochemical changes that enable it to assume a mesenchymal cell phenotype. EMT is involved in embryo development, wound healing, tissue regeneration, organ fibrosis and has also been proposed as the critical mechanism for the acquisition of malignant phenotypes by epithelial cancer cells. These cells have been shown to acquire a mesenchymal phenotype when localized at the invasive front of primary tumours increasing aggressiveness, invasiveness, metastatic potential and resistance to chemotherapy. There is now increasing evidence demonstrating that a crucial role in the development of this process is played by factors secreted by cells of the tumour microenvironment or by the tumour cells themselves. This review summarises the current knowledge of EMT induction in cancer by paracrine or autocrine mechanisms, by exosomes or free proteins and miRNAs.
- Published
- 2018
23. The metabolic landscape of cancer stem cells
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Marco Cordani, Massimo Donadelli, Elisa Dalla Pozza, Marta Palmieri, Ilaria Dando, and Giulia Biondani
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medicine.medical_treatment ,Clinical Biochemistry ,Cell Biology ,Biology ,Biochemistry ,Warburg effect ,Molecular oncology ,Cell biology ,Radiation therapy ,Cancer stem cell ,Cancer cell ,Genetics ,Cancer research ,medicine ,Molecular Biology - Abstract
Cancer stem cells (CSCs) are a sub-population of quiescent cells endowed with self-renewal properties that can sustain the malignant behavior of the tumor mass giving rise to more differentiated cancer cells. For this reason, the specific killing of CSCs represents one of the most important challenges of the modern molecular oncology. However, their particular resistance to traditional chemotherapy and radiotherapy imposes a thorough understanding of their biological and biochemical features. The metabolic peculiarities of CSCs may be a therapeutic and diagnostic opportunity in cancer research. In this review, we summarize the most significant discoveries on the metabolism of CSCs describing and critically analyzing the studies supporting either glycolysis or mitochondrial oxidative phosphorylation as a primary source of energy for CSCs.
- Published
- 2015
24. Mitochondrial uncoupling protein 2 and pancreatic cancer: A new potential target therapy
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Elisa Dalla Pozza, Massimo Donadelli, Ilaria Dando, and Marta Palmieri
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medicine.medical_specialty ,medicine.medical_treatment ,pancreatic cancer ,Antineoplastic Agents ,Biology ,Target therapy ,Ion Channels ,Metastasis ,Mitochondrial Proteins ,Internal medicine ,Pancreatic cancer ,Biomarkers, Tumor ,medicine ,Animals ,Humans ,Cytotoxic T cell ,Molecular Targeted Therapy ,chemistry.chemical_classification ,Reactive oxygen species ,Chemotherapy ,Uncoupling protein 2 ,Metabolism ,Gastroenterology ,Cancer ,Minireviews ,General Medicine ,medicine.disease ,Up-Regulation ,Pancreatic Neoplasms ,Endocrinology ,chemistry ,Anaerobic glycolysis ,Cancer cell ,Cancer research ,Signal Transduction - Abstract
Overall 5-years survival of pancreatic cancer patients is nearly 5%, making this cancer type one of the most lethal neoplasia. Furthermore, the incidence rate of pancreatic cancer has a growing trend that determines a constant increase in the number of deceases caused by this pathology. The poor prognosis of pancreatic cancer is mainly caused by delayed diagnosis, early metastasis of tumor, and resistance to almost all tested cytotoxic drugs. In this respect, the identification of novel potential targets for new and efficient therapies should be strongly encouraged in order to improve the clinical management of pancreatic cancer. Some studies have shown that the mitochondrial uncoupling protein 2 (UCP2) is over-expressed in pancreatic cancer as compared to adjacent normal tissues. In addition, recent discoveries established a key role of UCP2 in protecting cancer cells from an excessive production of mitochondrial superoxide ions and in the promotion of cancer cell metabolic reprogramming, including aerobic glycolysis stimulation, promotion of cancer progression. These observations together with the demonstration that UCP2 repression can synergize with standard chemotherapy to inhibit pancreatic cancer cell growth provide the molecular rationale to consider UCP2 as a potential therapeutic target for pancreatic cancer. In this editorial, recent advances describing the relationship between cancer development and mitochondrial UCP2 activity are critically provided.
- Published
- 2015
25. Trichostatin A alters cytoskeleton and energy metabolism of pancreatic adenocarcinoma cells: An in depth proteomic study
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Marcello Manfredi, Jessica Brandi, Massimo Donadelli, Eleonora Conte, Emilio Marengo, Arianna Buzzi, Raffaella Pacchiana, Daniela Cecconi, and Elisa Dalla Pozza
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0301 basic medicine ,Proteomics ,pancreatic cancer ,Biology ,trichostatin A ,Hydroxamic Acids ,Biochemistry ,03 medical and health sciences ,Pancreatic cancer ,Cell Line, Tumor ,medicine ,Humans ,2-deoxy-D-glucose ,cytoskeleton ,metabolism ,shotgun proteomics ,Shotgun proteomics ,Molecular Biology ,Cytoskeleton ,Cell growth ,Cell Biology ,medicine.disease ,Molecular biology ,Pancreatic Neoplasms ,030104 developmental biology ,Trichostatin A ,Mechanism of action ,Cell culture ,Cancer research ,Adenocarcinoma ,Histone deacetylase ,medicine.symptom ,Energy Metabolism ,medicine.drug ,Carcinoma, Pancreatic Ductal - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal of all human cancers with a high mortality rate. Resistance to conventional treatments and chemotherapeutics is a typical feature of PDAC. To investigate the causes of drug resistance it is essential to deeply investigate the mechanism of action of chemotherapeutics. In this study, we performed an in depth shotgun proteomic approach using the label-free proteomic SWATH-MS analysis to investigate novel insights of the mechanism of action of the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) in PDAC cells. This proteomic analysis in PaCa44 cells and data elaboration of TSA-regulated proteins by bioinformatics showed an overall up-regulation of cytokeratins and other proteins related to the cytoskeleton organization, keratinization, and apoptotic cell death. On the contrary, a large amount of the down-regulated proteins by TSA treatment belongs to the cellular energetic metabolism and to the machinery of protein synthesis, such as ribosomal proteins, determining synergistic cell growth inhibition by the combined treatment of TSA and the glycolytic inhibitor 2-deoxy-d-glucose in a panel of PDAC cell lines. Data are available via ProteomeXchange with identifier PXD007801.
- Published
- 2017
26. UCP2 inhibition induces ROS/Akt/mTOR axis: role of GAPDH nuclear translocation in genipin/everolimus anticancer synergism
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Ivana Cataldo, Stefano Bruno, Massimo Donadelli, Elisa Dalla Pozza, Marta Palmieri, Raffaella Pacchiana, Michele Caraglia, Anna Grimaldi, Ilaria Dando, Paola Conti, Aldo Scarpa, Marco Cordani, Giovanna Butera, Università degli Studi di Verona, Associazione Italiana per la Ricerca sul Cancro, Fondazione Umberto Veronesi, Dando, I, Pacchiana, R, Pozza, Ed, Cataldo, I, Bruno, S, Conti, P, Cordani, M, Grimaldi, A, Butera, G, Caraglia, M, Scarpa, A, Palmieri, M, and Donadelli, M.
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Male ,0301 basic medicine ,Apoptosis ,Pharmacology ,Biochemistry ,chemistry.chemical_compound ,Iridoids ,Uncoupling Protein 2 ,Pancreas cancer ,GAPDH ,TOR Serine-Threonine Kinases ,Glyceraldehyde-3-Phosphate Dehydrogenases ,Protein Transport ,Everolimu ,cell death ,mTOR ,Female ,Carcinoma, Pancreatic Ductal ,Signal Transduction ,medicine.drug ,Cell death ,UCP2 ,Programmed cell death ,uncoupling proteins ,Antineoplastic Agents ,everolimus ,pancreas cancer ,Biology ,03 medical and health sciences ,Cell Line, Tumor ,Physiology (medical) ,Uncoupling protein ,medicine ,Humans ,Everolimus ,Protein kinase B ,PI3K/AKT/mTOR pathway ,Cell Proliferation ,Cell growth ,RPTOR ,Autophagy ,Xenograft Model Antitumor Assays ,Pancreatic Neoplasms ,030104 developmental biology ,chemistry ,Genipin ,Uncoupling proteins ,Reactive Oxygen Species ,Proto-Oncogene Proteins c-akt - Abstract
Several studies indicate that mitochondrial uncoupling protein 2 (UCP2) plays a pivotal role in cancer development by decreasing reactive oxygen species (ROS) produced by mitochondrial metabolism and by sustaining chemoresistance to a plethora of anticancer drugs. Here, we demonstrate that inhibition of UCP2 triggers Akt/mTOR pathway in a ROS-dependent mechanism in pancreatic adenocarcinoma cells. This event reduces the antiproliferative outcome of UCP2 inhibition by genipin, creating the conditions for the synergistic counteraction of cancer cell growth with the mTOR inhibitor everolimus. Inhibition of pancreatic adenocarcinoma cell growth and induction of apoptosis by genipin and everolimus treatment are functionally related to nuclear translocation of the cytosolic glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH). The synthetic compound (S)-benzyl-2-amino-2-(S)-3-bromo-4,5-dihydroisoxazol-5-yl-acetate (AXP3009), which binds GAPDH at its redox-sensitive Cys152, restores cell viability affected by the combined treatment with genipin and everolimus, suggesting a role for ROS production in the nuclear translocation of GAPDH. Caspase-mediated apoptosis by genipin and everolimus is further potentiated by the autophagy inhibitor 3-methyladenine revealing a protective role for Beclin1-mediated autophagy induced by the treatment. Mice xenograft of pancreatic adenocarcinoma further confirmed the antiproliferative outcome of drug combination without toxic effects for animals. Tumor masses from mice injected with UCP2 and mTOR inhibitors revealed a strong reduction in tumor volume and number of mitosis associated with a marked GAPDH nuclear positivity. Altogether, these results reveal novel mechanisms through which UCP2 promotes cancer cell proliferation and support the combined inhibition of UCP2 and of Akt/mTOR pathway as a novel therapeutic strategy in the treatment of pancreatic adenocarcinoma., This work was supported by Joint Projects program 2015 from University of Verona to M. Donadelli (no. B12I15002320003) and by Associazione Italiana Ricerca Cancro (AIRC 12182) to A. Scarpa. Ilaria Dando is a fellow of Fondazione Umberto Veronesi. Elisa Dalla Pozza is a fellow of AIRC 5 per mille (Grant no. 12182).
- Published
- 2017
27. Corrigendum: Extracellular matrix composition modulates <scp>PDAC</scp> parenchymal and stem cell plasticity and behavior through the secretome
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Giulia Biondani, Katrine Zeeberg, Maria Raffaella Greco, Stefania Cannone, Ilaria Dando, Elisa Dalla Pozza, Maria Mastrodonato, Stefania Forciniti, Valeria Casavola, Marta Palmieri, Stephan Joel Reshkin, and Rosa Angela Cardone
- Subjects
Cell Biology ,Molecular Biology ,Biochemistry - Published
- 2019
28. Role of mitochondrial uncoupling protein 2 in cancer cell resistance to gemcitabine
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Chiara Costanzo, Claudia Fiorini, Anna Sgarbossa, Elisa Dalla Pozza, Massimo Donadelli, Marta Menegazzi, Marta Palmieri, and Ilaria Dando
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uncoupling protein 2 ,endocrine system diseases ,Uncoupling Agents ,pancreatic cancer ,Biology ,medicine.disease_cause ,Deoxycytidine ,Ion Channels ,Mitochondrial Proteins ,resistance ,Superoxides ,Cell Line, Tumor ,Neoplasms ,medicine ,Humans ,oxidative stress ,Gene silencing ,Iridoids ,RNA, Messenger ,Molecular Biology ,Cell Proliferation ,Cancer ,chemistry.chemical_classification ,Reactive oxygen species ,Superoxide Dismutase ,gemcitabine ,apoptosis ,Cell Biology ,Acetylcysteine ,Cell biology ,Gene Expression Regulation, Neoplastic ,chemistry ,Biochemistry ,Drug Resistance, Neoplasm ,Mitochondrial matrix ,Cell culture ,Apoptosis ,Cancer cell ,Poly(ADP-ribose) Polymerases ,Oxidative stress - Abstract
Cancer cells exhibit an endogenous constitutive oxidative stress higher than that of normal cells, which renders tumours vulnerable to further reactive oxygen species (ROS) production. Mitochondrial uncoupling protein 2 (UCP2) can mitigate oxidative stress by increasing the influx of protons into the mitochondrial matrix and reducing electron leakage and mitochondrial superoxide generation. Here, we demonstrate that chemical uncouplers or UCP2 over-expression strongly decrease mitochondrial superoxide induction by the anticancer drug gemcitabine (GEM) and protect cancer cells from GEM-induced apoptosis. Moreover, we show that GEM IC50 values well correlate with the endogenous level of UCP2 mRNA, suggesting a critical role for mitochondrial uncoupling in GEM resistance. Interestingly, GEM treatment stimulates UCP2 mRNA expression suggesting that mitochondrial uncoupling could have a role also in the acquired resistance to GEM. Conversely, UCP2 inhibition by genipin or UCP2 mRNA silencing strongly enhances GEM-induced mitochondrial superoxide generation and apoptosis, synergistically inhibiting cancer cell proliferation. These events are significantly reduced by the addition of the radical scavenger N-acetyl-l-cysteine or MnSOD over-expression, demonstrating a critical role of the oxidative stress. Normal primary fibroblasts are much less sensitive to GEM/genipin combination. Our results demonstrate for the first time that UCP2 has a role in cancer cell resistance to GEM supporting the development of an anti-cancer therapy based on UCP2 inhibition associated to GEM treatment.
- Published
- 2012
29. Proteomic analysis of pancreatic cancer stem cells: Functional role of fatty acid synthesis and mevalonate pathways
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Kevin Park, Lello Zolla, Jessica Brandi, Marta Palmieri, Daniela Cecconi, Aldo Scarpa, Giulia Biondani, Rosalind E. Jenkins, Giuseppina Fanelli, Elisa Dalla Pozza, Ilaria Dando, Eithne Costello, and Victoria Elliott
- Subjects
0301 basic medicine ,cancer stem cells ,Proteomics ,pancreatic cancer ,Biophysics ,pancreatic ductal adenocarcinoma ,Mevalonic Acid ,Biology ,Biochemistry ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Downregulation and upregulation ,Cancer stem cell ,Pancreatic cancer ,Cell Line, Tumor ,medicine ,Humans ,Metabolomics ,Glycolysis ,Pancreas ,Fatty acid synthesis ,PDAC ,Fatty Acids ,medicine.disease ,Cell biology ,Cerulenin ,Pancreatic Neoplasms ,Metabolic pathway ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,Metabolome ,Neoplastic Stem Cells ,Stem cell ,Fatty Acid Synthases ,Metabolic Networks and Pathways ,Carcinoma, Pancreatic Ductal - Abstract
Recently, we have shown that the secretome of pancreatic cancer stem cells (CSCs) is characterized by proteins that participate in cancer differentiation, invasion, and metastasis. However, the differentially expressed intracellular proteins that lead to the specific characteristics of pancreatic CSCs have not yet been identified, and as a consequence the deranged metabolic pathways are yet to be elucidated. To identify the modulated proteins of pancreatic CSCs, iTRAQ-based proteomic analysis was performed to compare the proteome of Panc1 CSCs and Panc1 parental cells, identifying 230 modulated proteins. Pathway analysis revealed activation of glycolysis, the pentose phosphate pathway, the pyruvate-malate cycle, and lipid metabolism as well as downregulation of the Krebs cycle, the splicesome and non-homologous end joining. These findings were supported by metabolomics and immunoblotting analysis. It was also found that inhibition of fatty acid synthase by cerulenin and of mevalonate pathways by atorvastatin have a greater anti-proliferative effect on cancer stem cells than parental cells. Taken together, these results clarify some important aspects of the metabolic network signature of pancreatic cancer stem cells, shedding light on key and novel therapeutic targets and suggesting that fatty acid synthesis and mevalonate pathways play a key role in ensuring their viability. Biological significance To better understand the altered metabolic pathways of pancreatic cancer stem cells (CSCs), a comprehensive proteomic analysis and metabolite profiling investigation of Panc1 and Panc1 CSCs were carried out. The findings obtained indicate that Panc1 CSCs are characterized by upregulation of glycolysis, pentose phosphate pathway, pyruvate-malate cycle, and lipid metabolism and by downregulation of Krebs cycle, spliceosome and non-homologous end joining. Moreover, fatty acid synthesis and mevalonate pathways are shown to play a critical contribution to the survival of pancreatic cancer stem cells. This study is helpful for broadening the knowledge of pancreatic cancer stem cells and could accelerate the development of novel therapeutic strategies.
- Published
- 2016
30. The antioxidant uncoupling protein 2 stimulates hnRNPA2/B1, GLUT1 and PKM2 expression and sensitizes pancreas cancer cells to glycolysis inhibition
- Author
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Elisa Dalla Pozza, Jordi Oliver, Jessica Brandi, Massimo Donadelli, Emilio Marengo, Raffaella Pacchiana, Marcello Manfredi, Ilaria Dando, Daniela Cecconi, Margalida Torrens-Mas, Giovanna Butera, Marco Cordani, and Pilar Roca
- Subjects
0301 basic medicine ,Proteomics ,Biochemistry ,Oxidative Phosphorylation ,chemistry.chemical_compound ,Glycolysis Inhibition ,0302 clinical medicine ,Insulin-Secreting Cells ,Heterogeneous-Nuclear Ribonucleoprotein Group A-B ,Uncoupling protein ,Iridoids ,Uncoupling Protein 2 ,RNA, Small Interfering ,Cancer ,Glucose Transporter Type 1 ,biology ,Mitochondria ,Cell biology ,Gene Expression Regulation, Neoplastic ,030220 oncology & carcinogenesis ,Cancer, Metabolism, Uncoupling protein,s UCP2, Warburg effect, Proteomics ,s UCP2 ,Glycolysis ,Signal Transduction ,Thyroid Hormones ,Cellular respiration ,Oxidative phosphorylation ,Deoxyglucose ,PKM2 ,03 medical and health sciences ,Cell Line, Tumor ,Physiology (medical) ,Lactate dehydrogenase ,Humans ,Cytochrome c oxidase ,RNA, Messenger ,Cell Proliferation ,L-Lactate Dehydrogenase ,Gene Expression Profiling ,Membrane Proteins ,Acetylcysteine ,030104 developmental biology ,Metabolism ,chemistry ,Cancer cell ,biology.protein ,Warburg effect ,Carrier Proteins ,Reactive Oxygen Species - Abstract
Several evidence indicate that metabolic alterations play a pivotal role in cancer development. Here, we report that the mitochondrial uncoupling protein 2 (UCP2) sustains the metabolic shift from mitochondrial oxidative phosphorylation (mtOXPHOS) to glycolysis in pancreas cancer cells. Indeed, we show that UCP2 sensitizes pancreas cancer cells to the treatment with the glycolytic inhibitor 2-deoxy-D-glucose. Through a bidimensional electrophoresis analysis, we identify 19 protein species differentially expressed after treatment with the UCP2 inhibitor genipin and, by bioinformatic analyses, we show that these proteins are mainly involved in metabolic processes. In particular, we demonstrate that the antioxidant UCP2 induces the expression of hnRNPA2/B1, which is involved in the regulation of both GLUT1 and PKM2 mRNAs, and of lactate dehydrogenase (LDH) increasing the secretion of L-lactic acid. We further demonstrate that the radical scavenger N-acetyl-L-cysteine reverts hnRNPA2/B1 and PKM2 inhibition by genipin indicating a role for reactive oxygen species in the metabolic reprogramming of cancer cells mediated by UCP2. We also observe an UCP2-dependent decrease in mtOXPHOS complex I (NADH dehydrogenase), complex IV (cytochrome c oxidase), complex V (ATPase) and in mitochondrial oxygen consumption, suggesting a role for UCP2 in the counteraction of pancreatic cancer cellular respiration. All these results reveal novel mechanisms through which UCP2 promotes cancer cell proliferation with the concomitant metabolic shift from mtOXPHOS to the glycolytic pathway.
- Published
- 2016
31. Secretome protein signature of human pancreatic cancer stem-like cells
- Author
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Jessica Brandi, Elisa Robotti, Marta Palmieri, Rosalind E. Jenkins, Aldo Scarpa, Elisa Dalla Pozza, Emilio Marengo, Eithne Costello, Kevin Park, Ilaria Dando, Victoria Elliott, Daniela Cecconi, and Giulia Biondani
- Subjects
0301 basic medicine ,Proteomics ,Proteome ,Biophysics ,stem-cell ,Biology ,Bioinformatics ,Biochemistry ,Metastasis ,Cell Line ,03 medical and health sciences ,0302 clinical medicine ,Cancer stem cell ,Pancreatic cancer ,medicine ,Humans ,MARCKS ,Pancreas ,Secretome ,Stem Cells ,Cancer ,Secretomics ,medicine.disease ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Cancer research - Abstract
Emerging research has demonstrated that pancreatic ductal adenocarcinoma (PDAC) contains a sub-population of cancer stem cells (CSCs) characterized by self-renewal, anchorage-independent-growth, long-term proliferation and chemoresistance. The secretome analysis of pancreatic CSCs has not yet been performed, although it may provide insight into tumour/microenvironment interactions and intracellular processes, as well as to identify potential biomarkers. To characterize the secreted proteins of pancreatic CSCs, we performed an iTRAQ-based proteomic analysis to compare the secretomes of Panc1 cancer stem-like cells (Panc1 CSCs) and parental cell line. A total of 72 proteins were found up-/down-regulated in the conditioned medium of Panc1 CSCs. The pathway analysis revealed modulation of vital physiological pathways including glycolysis, gluconeogenesis and pentose phosphate. Through ELISA immunoassays we analysed the presence of the three proteins most highly secreted by Panc1 CSCs (ceruloplasmin, galectin-3, and MARCKS) in sera of PDAC patient. ROC curve analysis suggests ceruloplasmin as promising marker for patients negative for CA19-9. Overall, our study provides a systemic secretome analysis of pancreatic CSCs revealing a number of secreted proteins which participate in pathological conditions including cancer differentiation, invasion and metastasis. They may serve as a valuable pool of proteins from which biomarkers and therapeutic targets can be identified. Biological significance The secretome of CSCs is a rich reservoir of biomarkers of cancer progression and molecular therapeutic targets, and thus is a topic of great interest for cancer research. The secretome analysis of pancreatic CSCs has not yet been performed. Recently, our group has demonstrated that Panc-1 CSCs isolated from parental cell line by using the CSC selective medium, represent a model of great importance to deepen the understanding of the biology of pancreatic adenocarcinoma. To our knowledge, this is the first proteomic study of pancreatic CSC secretome. We performed an iTRAQ-based analysis to compare the secretomes of Panc1 CSCs and Panc1 parental cell line and identified a total of 43 proteins secreted at higher level by pancreatic cancer stem cells. We found modulation of different vital physiological pathways (such as glycolysis and gluconeogenesis, pentose phosphate pathway) and the involvement of CSC secreted proteins (for example 72 kDa type IV collagenase, galectin-3, alpha-actinin-4, and MARCKS) in pathological conditions including cancer differentiation, invasion and metastasis. By ELISA verification we found that MARCKS and ceruloplasmin discriminate between controls and PDAC patients; in addition ROC curve analyses indicate that MARCKS does not have diagnostic accuracy, while ceruloplasmin could be a promising marker only for patients negative for CA19-9. We think that the findings reported in our manuscript advance the understanding of the pathways implicated in tumourigenesis, metastasis and chemoresistance of pancreatic cancer, and also identify a pool of proteins from which novel candidate diagnostic and therapeutic biomarkers could be discovered.
- Published
- 2016
32. Synergistic effect of trichostatin A and 5-aza-2′-deoxycytidine on growth inhibition of pancreatic endocrine tumour cell lines: A proteomic study
- Author
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Elisa Dalla Pozza, Maria Teresa Scupoli, Lello Zolla, Pier Giorgio Righetti, Sara Rinalducci, Marta Palmieri, Massimo Donadelli, Daniela Cecconi, and Aldo Scarpa
- Subjects
Pancreatic disease ,DNA Methyltransferase Inhibitor ,Apoptosis ,Enteroendocrine cell ,Biology ,Decitabine ,Hydroxamic Acids ,Biochemistry ,Pancreatic tumor ,Cell Line, Tumor ,Endocrine Gland Neoplasms ,medicine ,Humans ,Enzyme Inhibitors ,Molecular Biology ,Endocrine gland neoplasm ,Cell Proliferation ,Analysis of Variance ,Histone deacetylase inhibitor ,Cell growth ,Gene Expression Profiling ,Cell Cycle ,Drug Synergism ,Cell cycle ,medicine.disease ,Gene Expression Regulation, Neoplastic ,Pancreatic Neoplasms ,Trichostatin A ,Pancreatic endocrine tumour ,Azacitidine ,Cancer research ,Epigenetics ,Carcinoma, Pancreatic Ductal ,DNA methyl transferase inhibitor ,medicine.drug - Abstract
Our research group recently reported that pancreatic endocrine cancer cell lines are sensitive to the HDAC inhibitor trichostatin A (TSA). In the present paper, we show that the combined treatment of pancreatic endocrine tumour cell lines with TSA and the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (DAC) determines a strong synergistic inhibition of proliferation mainly due to apoptotic cell death. Proteomic analysis demonstrates that the modulation of specific proteins correlates with the antiproliferative effect of the drugs. A schematic network clarifies the most important targets or pathways involved in pancreatic endocrine cancer growth inhibition by single or combined drug treatments, which include proteasome, mitochondrial apoptotic pathway and caspase related proteins, p53 and Ras related proteins. A comparison between the patterns of proteins regulated by TSA or DAC in endocrine and ductal pancreatic cancer cell lines is also presented.
- Published
- 2009
33. Pancreatic ductal adenocarcinoma cell lines display a plastic ability to bi‑directionally convert into cancer stem cells
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Elisa Zoratti, Giulia Biondani, Elisa Dalla Pozza, Daniela Cecconi, Federico Boschi, Marta Palmieri, Davide Melisi, Jessica Brandi, Chiara Costanzo, Aldo Scarpa, Ilaria Dando, Maria Teresa Scupoli, and Matteo Fassan
- Subjects
Oncology ,Cancer Research ,medicine.medical_specialty ,cancer stem cell ,chemotherapy resistance ,endocrine system diseases ,Cell ,Mice, Nude ,pancreatic ductal adenocarcinoma ,Biology ,medicine.disease_cause ,Metastasis ,Mice ,Cancer stem cell ,Cell Line, Tumor ,Internal medicine ,Biomarkers, Tumor ,medicine ,Animals ,Humans ,metastasis ,Cells, Cultured ,Cell Differentiation ,Cell Dedifferentiation ,Cell cycle ,medicine.disease ,digestive system diseases ,Gene Expression Regulation, Neoplastic ,Pancreatic Neoplasms ,medicine.anatomical_structure ,Cancer cell ,Neoplastic Stem Cells ,Cancer research ,Adenocarcinoma ,Female ,Stem cell ,Carcinogenesis ,Carcinoma, Pancreatic Ductal - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is often diagnosed when metastatic events have occurred. Cancer stem cells (CSCs) play an important role in tumor initiation, metastasis, chemoresistance and relapse. A growing number of studies have suggested that CSCs exist in a dynamic equilibrium with more differentiated cancer cells via a bi‑directional regeneration that is dependent on the environmental stimuli. In this investigation, we obtain, by using a selective medium, PDAC CSCs from five out of nine PDAC cell lines, endowed with different tumorsphere‑forming ability. PDAC CSCs were generally more resistant to the action of five anticancer drugs than parental cell lines and were characterized by an increased expression of EpCAM and CD44v6, typical stem cell surface markers, and a decreased expression of E‑cadherin, the main marker of the epithelial state. PDAC CSCs were able to re‑differentiate into parental cells once cultured in parental growth condition, as demonstrated by re‑acquisition of the epithelial morphology, the decreased expression levels of EpCAM and CD44v6 and the increased sensitivity to anticancer drugs. Finally, PDAC CSCs injected into nude mice developed a larger subcutaneous tumor mass and showed a higher metastatic activity compared to parental cells. The present study demonstrates the ability to obtain CSCs from several PDAC cell lines and that these cells are differentially resistant to various anticancer agents. This variability renders them a model of great importance to deeply understand pancreatic adenocarcinoma biology, to discover new biomarkers and to screen new therapeutic compounds.
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- 2015
34. Antioxidant Mechanisms and ROS-Related MicroRNAs in Cancer Stem Cells
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Elisa Dalla Pozza, Massimo Donadelli, Marta Palmieri, Marco Cordani, Ilaria Dando, and Giulia Biondani
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Aging ,cancer stem cell ,Cellular differentiation ,Population ,Review Article ,Biology ,Biochemistry ,Antioxidants ,Metastasis ,Cancer stem cell ,Neoplasms ,microRNA ,medicine ,Biomarkers, Tumor ,Humans ,lcsh:QH573-671 ,education ,chemistry.chemical_classification ,Reactive oxygen species ,education.field_of_study ,lcsh:Cytology ,Cancer ,ROS ,Cell Differentiation ,Cell Biology ,General Medicine ,medicine.disease ,Cell biology ,MicroRNAs ,chemistry ,cancer stem cell, microRNA, ROS ,Cancer cell ,Neoplastic Stem Cells ,Reactive Oxygen Species - Abstract
Increasing evidence indicates that most of the tumors are sustained by a distinct population of cancer stem cells (CSCs), which are responsible for growth, metastasis, invasion, and recurrence. CSCs are typically characterized by self-renewal, the key biological process allowing continuous tumor proliferation, as well as by differentiation potential, which leads to the formation of the bulk of the tumor mass. CSCs have several advantages over the differentiated cancer cell populations, including the resistance to radio- and chemotherapy, and their gene-expression programs have been shown to correlate with poor clinical outcome, further supporting the relevance of stemness properties in cancer. The observation that CSCs possess enhanced mechanisms of protection from reactive oxygen species (ROS) induced stress and a different metabolism from the differentiated part of the tumor has paved the way to develop drugs targeting CSC specific signaling. In this review, we describe the role of ROS and of ROS-related microRNAs in the establishment and maintenance of self-renewal and differentiation capacities of CSCs.
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- 2014
35. Hyaluronic acid-coated liposomes for active targeting of gemcitabine
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Nicolas Tsapis, Carlotta Lerda, Barbara Stella, Chiara Costanzo, Elisa Dalla Pozza, Massimo Donadelli, Luigi Cattel, Elias Fattal, Silvia Arpicco, Marta Palmieri, and Ilaria Dando
- Subjects
Antimetabolites, Antineoplastic ,Phospholipid ,Pharmaceutical Science ,Adenocarcinoma ,Deoxycytidine ,chemistry.chemical_compound ,Drug Delivery Systems ,Cell surface receptor ,Cell Line, Tumor ,Hyaluronic acid ,Humans ,Prodrugs ,Hyaluronic Acid ,Cytotoxicity ,liposomes ,gemcitabine ,Cancer ,Cellular localization ,Liposome ,Chemistry ,Cell growth ,Phosphatidylethanolamines ,General Medicine ,Ligand (biochemistry) ,Gene Expression Regulation, Neoplastic ,Molecular Weight ,Pancreatic Neoplasms ,Hyaluronan Receptors ,Biochemistry ,Liposomes ,Biotechnology - Abstract
The aim of this work was the preparation, characterization, and preliminary evaluation of the targeting ability toward pancreatic adenocarcinoma cells of liposomes containing the gemcitabine lipophilic prodrug [4-(N)-lauroyl-gemcitabine, C12GEM]. Hyaluronic acid (HA) was selected as targeting agent since it is biodegradable, biocompatible, and can be chemically modified and its cell surface receptor CD44 is overexpressed on various tumors. For this purpose, conjugates between a phospholipid, the 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE), and HA of two different low molecular weights 4800 Da (12 disaccharidic units) and 12,000 Da (32 disaccharidic units), were prepared, characterized, and introduced in the liposomes during the preparation. Different liposomal formulations were prepared and their characteristics were analyzed: size, Z potential, and TEM analyses underline a difference in the HA-liposomes from the non-HA ones. In order to better understand the HA-liposome cellular localization and to evaluate their interaction with CD44 receptor, confocal microscopy studies were performed. The results demonstrate that HA facilitates the recognition of liposomes by MiaPaCa2 cells (CD44(+)) and that the uptake increases with increase in the polymer molecular weight. Finally, the cytotoxicity of the different preparations was evaluated and data show that incorporation of C12GEM increases their cytotoxic activity and that HA-liposomes inhibit cell growth more than plain liposomes. Altogether, the results demonstrate the specificity of C12GEM targeting toward CD44-overexpressing pancreatic adenocarcinoma cell line using HA as a ligand.
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- 2013
36. UCP2 inhibition triggers ROS-dependent nuclear translocation of GAPDH and autophagic cell death in pancreatic adenocarcinoma cells
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Elisa Dalla Pozza, Massimo Donadelli, Chiara Costanzo, Claudia Fiorini, Marta Palmieri, Ilaria Dando, and Chiara Padroni
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Programmed cell death ,Cholagogues and Choleretics ,Blotting, Western ,Respiratory chain ,Fluorescent Antibody Technique ,Apoptosis ,Adenocarcinoma ,medicine.disease_cause ,Real-Time Polymerase Chain Reaction ,Ion Channels ,Mitochondrial Proteins ,ucp2 ,Cell Line, Tumor ,Genipin ,medicine ,Autophagy ,Uncoupling protein ,Humans ,Iridoids ,Uncoupling Protein 2 ,RNA, Messenger ,RNA, Small Interfering ,Molecular Biology ,ROS ,GAPDH ,Cancer ,Glyceraldehyde 3-phosphate dehydrogenase ,Cell Proliferation ,chemistry.chemical_classification ,Reactive oxygen species ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,Glyceraldehyde-3-Phosphate Dehydrogenases ,Cell Biology ,Cell biology ,Pancreatic Neoplasms ,Oxidative Stress ,Protein Transport ,chemistry ,biology.protein ,Cancer research ,Reactive Oxygen Species ,Oxidative stress - Abstract
Mitochondrial uncoupling protein 2 (UCP2) can moderate oxidative stress by favoring the influx of protons into the mitochondrial matrix, thus reducing electron leakage from respiratory chain and mitochondrial superoxide production. Here, we demonstrate that UCP2 inhibition by genipin or UCP2 siRNA strongly increases reactive oxygen species (ROS) production inhibiting pancreatic adenocarcinoma cell growth. We also show that UCP2 inhibition triggers ROS-dependent nuclear translocation of the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH), formation of autophagosomes, and the expression of the autophagy marker LC3-II. Consistently, UCP2 over-expression significantly reduces basal autophagy confirming the anti-autophagic role of UCP2. Furthermore, we demonstrate that autophagy induced by UCP2 inhibition determines a ROS-dependent cell death, as indicated by the apoptosis decrease in the presence of the autophagy inhibitors chloroquine (CQ) or 3-methyladenine (3-MA), or the radical scavenger NAC. Intriguingly, the autophagy induced by genipin is able to potentiate the autophagic cell death triggered by gemcitabine, the standard chemotherapeutic drug for pancreatic adenocarcinoma, supporting the development of an anti-cancer therapy based on UCP2 inhibition associated to standard chemotherapy. Our results demonstrate for the first time that UCP2 plays a role in autophagy regulation bringing new insights into mitochondrial uncoupling protein field.
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- 2012
37. Expression of the Antiapoptotic Protein BAG3 Is a Feature of Pancreatic Adenocarcinoma and Its Overexpression Is Associated With Poorer Survival. Turco MC: Corresponding Author
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Rosati, Alessandra, Samantha, Bersani, Francesca, Tavano, Elisa Dalla Pozza, DE MARCO, Margot, Marta, Palmieri, Vincenzo De Laurenzi, Renato, Franco, Giosuè, Scognamiglio, Raffaele, Palaia, Andrea, Fontana, Pierluigi di Sebastiano, Massimo, Donadelli, Ilaria, Dando, Jan Paul Medema, Frederike, Dijk, Lieke, Welling, Fabio Francesco di Mola, Raffaele, Pezzilli, Turco, Maria Caterina, and Aldo, Scarpa
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- 2012
38. HYALURONIC ACID-MODIFIED LIPOSOMES: PREPARATION, CHARACTERIZATION AND BIOLOGICAL EVALUATION
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Arpicco, Silvia Maria, Carlotta, Lerda, Elisa Dalla Pozza, Chiara, Costanzo, Stella, Barbara, Cattel, Luigi, and Marta, Palmieri
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- 2012
39. Gemcitabine response in pancreatic adenocarcinoma cells is synergistically enhanced by dithiocarbamate derivatives
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Elisa Dalla Pozza, Massimo Donadelli, Tatyana Zaniboni, Marta Franchini, Chiara Costanzo, Silvia Arpicco, Marta Palmieri, Aldo Scarpa, and Ilaria Dando
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endocrine system diseases ,Blotting, Western ,Apoptosis ,Adenocarcinoma ,Biochemistry ,pancreatic adenocarcinoma ,gemcitabine ,zinc ,dithiocarbamate ,oxidative stress ,apoptosis ,Deoxycytidine ,chemistry.chemical_compound ,Pyrrolidine dithiocarbamate ,In vivo ,Thiocarbamates ,Physiology (medical) ,Pancreatic cancer ,Cell Line, Tumor ,medicine ,Humans ,Cell Proliferation ,Cell growth ,Drug Synergism ,medicine.disease ,Gemcitabine ,Pancreatic Neoplasms ,chemistry ,Cell culture ,Cancer research ,medicine.drug - Abstract
Pancreatic adenocarcinoma is a common malignancy that remains refractory to all available therapies, including the gold standard drug gemcitabine (GEM). We investigated the effect of the combination of GEM and each of the ionophore compounds pyrrolidine dithiocarbamate (PDTC) and disulfiram [DSF; 1-(diethylthiocarbamoyldisulfanyl)-N,N-diethylmethanethioamide] on p53(-/-) pancreatic adenocarcinoma cell growth. PDTC or DSF synergistically inhibited cell proliferation when used in combination with GEM by inducing apoptotic cell death. This effect was associated with an increased mitochondrial O(2)(•-) production and was further enhanced by zinc ions. Basal levels of mitochondrial O(2)(•-) or manganese superoxide dismutase (MnSOD) strictly correlated with the IC(50) for GEM or the percentage of synergism. Thus, the most relevant values of the antiproliferative synergism were obtained in GEM-resistant pancreatic adenocarcinoma cell lines. Interestingly, the GEM-sensitive T3M4 cells transfected with MnSOD expression vector showed mitochondrial O(2)(•-) and IC(50) for GEM similar to those of resistant cell lines. In vivo experiments performed on nude mice xenotransplanted with the GEM-resistant PaCa44 cell line showed that only the combined treatment with GEM and DSF/Zn completely inhibited the growth of the tumoral masses. These results and the consideration that DSF is already used in clinics strongly support the GEM and DSF/Zn combination as a new approach to overcoming pancreatic cancer resistance to standard chemotherapy.
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- 2010
40. Increased stability of P21(WAF1/CIP1) mRNA is required for ROS/ERK-dependent pancreatic adenocarcinoma cell growth inhibition by pyrrolidine dithiocarbamate
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Chiara Costanzo, Maria Teresa Scupoli, Massimo Donadelli, Paolo Piacentini, Marta Palmieri, Elisa Dalla Pozza, and Aldo Scarpa
- Subjects
MAPK/ERK pathway ,p53 ,Pyrrolidines ,RNA Stability ,Oligonucleotides ,chemistry.chemical_compound ,Mice ,Pyrrolidine dithiocarbamate ,oxidative stress ,p21WAF1/CIP1 ,Tumor ,pancreatic adenocarcinoma ,pyrrolidine dithiocarbamate ,ERK1/2 ,p21(WAF1/CIP1) ,Acetylcysteine ,Adenocarcinoma ,Animals ,Cell Cycle ,Cell Line, Tumor ,Cell Proliferation ,Cyclin-Dependent Kinase Inhibitor p21 ,DNA Primers ,Fibroblasts ,Flavonoids ,Gene Expression Regulation, Neoplastic ,Gene Silencing ,Humans ,Immunoblotting ,Oligonucleotides, Antisense ,Pancreatic Neoplasms ,Reactive Oxygen Species ,Reverse Transcriptase Polymerase Chain Reaction ,Thiocarbamates ,Cell cycle ,Free radical scavenger ,medicine.anatomical_structure ,Growth inhibition ,Biology ,Cell Line ,medicine ,Antisense ,Fibroblast ,Molecular Biology ,Neoplastic ,Cell growth ,Cell Biology ,Molecular biology ,chemistry ,Gene Expression Regulation ,Cancer cell - Abstract
We present evidence that pyrrolidine dithiocarbamate (PDTC) inhibits growth of p53-negative pancreatic adenocarcinoma cell lines via cell cycle arrest in the S-phase, while it has no effect on primary fibroblast proliferation. Growth inhibition of cancer cells is dependent on ROS and ERK1/2 induction as indicated by a significantly reduced PDTC-associated growth inhibition by the free radical scavenger N-acetyl-l-cysteine (NAC) or the MEK/ERK1/2 inhibitor (PD98059). Moreover, ERK1/2 induction is dependent on ROS production as demonstrated by a complete removal of PDTC-mediated ERK1/2 phosphorylation by NAC. p21WAF1/CIP1 activation has a central role in growth inhibition by PDTC, as revealed by P21WAF1/CIP1 silencing experiments with antisense oligonucleotide, and occurs via increased mRNA stability largely mediated by ROS/ERK induction. Conversely, PDTC does not affect P21WAF1/CIP1 gene expression in primary fibroblasts, although it is able to activate p53 and the p53-regulated antioxidant SESN2. These results suggest that the resistance of fibroblasts to the cytotoxic action of PDTC may be related to the up-regulation of p53-dependent antioxidant genes. Finally, in vivo studies on PaCa44 cells subcutaneously xenografted in nude mice show that treatment with 100 or 200 mg/kg PDTC reduces of 30% or 60% the tumour volume, respectively, and does not cause any apparent form of toxicity.
- Published
- 2006
41. Mitochondrial Elongation and OPA1 Play Crucial Roles during the Stemness Acquisition Process in Pancreatic Ductal Adenocarcinoma
- Author
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Cristian Andres Carmona-Carmona, Elisa Dalla Pozza, Giulia Ambrosini, Barbara Cisterna, Marta Palmieri, Ilaria Decimo, José M. Cuezva, Emanuela Bottani, Ilaria Dando, and UAM. Departamento de Biología Molecular
- Subjects
cancer stem cells ,Cancer Research ,Messenger RNA ,pancreatic ductal adenocarcinoma ,Small interfering RNA ,Biología y Biomedicina / Biología ,OPA1 ,mitochondrial dynamics ,mitochondrial fusion ,Mitochondrial DNA ,Dynamin ,Oncology - Abstract
Pancreatic ductal adenocarcinoma (PDAC) is the most common type of pancreatic cancer with an overall 5-year survival rate of less than 9%. The high aggressiveness of PDAC is linked to the presence of a subpopulation of cancer cells with a greater tumorigenic capacity, generically called cancer stem cells (CSCs). CSCs present a heterogeneous metabolic profile that might be supported by an adaptation of mitochondrial function; however, the role of this organelle in the development and maintenance of CSCs remains controversial. To determine the role of mitochondria in CSCs over longer periods, which may reflect more accurately their quiescent state, we studied the mitochondrial physiology in CSCs at short-, medium-, and long-term culture periods. We found that CSCs show a significant increase in mitochondrial mass, more mitochondrial fusion, and higher mRNA expression of genes involved in mitochondrial biogenesis than parental cells. These changes are accompanied by a regulation of the activities of OXPHOS complexes II and IV. Furthermore, the protein OPA1, which is involved in mitochondrial dynamics, is overexpressed in CSCs and modulates the tumorsphere formation. Our findings indicate that CSCs undergo mitochondrial remodeling during the stemness acquisition process, which could be exploited as a promising therapeutic target against pancreatic CSCs.
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42. Mitochondrial uncoupling protein 2 and pancreatic cancer: a new potential target therapy.
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Donadelli M, Dando I, Dalla Pozza E, and Palmieri M
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- Animals, Antineoplastic Agents therapeutic use, Biomarkers, Tumor antagonists & inhibitors, Humans, Ion Channels antagonists & inhibitors, Mitochondrial Proteins antagonists & inhibitors, Molecular Targeted Therapy, Pancreatic Neoplasms drug therapy, Pancreatic Neoplasms mortality, Pancreatic Neoplasms pathology, Uncoupling Protein 2, Up-Regulation, Biomarkers, Tumor metabolism, Ion Channels metabolism, Mitochondrial Proteins metabolism, Pancreatic Neoplasms metabolism, Signal Transduction drug effects
- Abstract
Overall 5-years survival of pancreatic cancer patients is nearly 5%, making this cancer type one of the most lethal neoplasia. Furthermore, the incidence rate of pancreatic cancer has a growing trend that determines a constant increase in the number of deceases caused by this pathology. The poor prognosis of pancreatic cancer is mainly caused by delayed diagnosis, early metastasis of tumor, and resistance to almost all tested cytotoxic drugs. In this respect, the identification of novel potential targets for new and efficient therapies should be strongly encouraged in order to improve the clinical management of pancreatic cancer. Some studies have shown that the mitochondrial uncoupling protein 2 (UCP2) is over-expressed in pancreatic cancer as compared to adjacent normal tissues. In addition, recent discoveries established a key role of UCP2 in protecting cancer cells from an excessive production of mitochondrial superoxide ions and in the promotion of cancer cell metabolic reprogramming, including aerobic glycolysis stimulation, promotion of cancer progression. These observations together with the demonstration that UCP2 repression can synergize with standard chemotherapy to inhibit pancreatic cancer cell growth provide the molecular rationale to consider UCP2 as a potential therapeutic target for pancreatic cancer. In this editorial, recent advances describing the relationship between cancer development and mitochondrial UCP2 activity are critically provided.
- Published
- 2015
- Full Text
- View/download PDF
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