153 results on '"Marco Tripodi"'
Search Results
2. Data from Design and Functional Validation of a Mutant Variant of the LncRNA HOTAIR to Counteract Snail Function in Epithelial-to-Mesenchymal Transition
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Carla Cicchini, Marco Tripodi, Gian Gaetano Tartaglia, Raffaele Strippoli, Andrea Vandelli, Laura Santangelo, Claudia Montaldo, Veronica Riccioni, Sabrina Garbo, and Cecilia Battistelli
- Abstract
HOTAIR is a lncRNA overexpressed in several epithelial cancers and strongly correlated with invasion. This lncRNA was proven a pivotal element of the epithelial-to-mesenchymal transition (EMT), a transdifferentiation process triggering metastasis. Snail, master inducer of EMT, requires HOTAIR to recruit EZH2 on specific epithelial target genes (i.e., HNF4α, E-cadherin, and HNF1α) and cause their repression. Here, we designed a HOTAIR deletion mutant form, named HOTAIR-sbid, including the putative Snail-binding domain but depleted of the EZH2-binding domain. HOTAIR-sbid acted as a dominant negative of the endogenous HOTAIR. In both murine and human tumor cells, HOTAIR-sbid impaired the ability of HOTAIR to bind Snail and, in turn, trigger H3K27me3/EZH2-mediated repression of Snail epithelial target genes. Notably, HOTAIR-sbid expression was proven to reduce cellular motility, invasiveness, anchorage-independent growth, and responsiveness to TGFβ-induced EMT. These data provide evidence on a lncRNA-based strategy to effectively impair the function of a master EMT-transcriptional factor.Significance:This study defines an innovative RNA-based strategy to interfere with a pivotal function of the tumor-related lncRNA HOTAIR, comprising a dominant negative mutant that was computationally designed and that impairs epithelial-to-mesenchymal transition.
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- 2023
3. Supplementary Data from Design and Functional Validation of a Mutant Variant of the LncRNA HOTAIR to Counteract Snail Function in Epithelial-to-Mesenchymal Transition
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Carla Cicchini, Marco Tripodi, Gian Gaetano Tartaglia, Raffaele Strippoli, Andrea Vandelli, Laura Santangelo, Claudia Montaldo, Veronica Riccioni, Sabrina Garbo, and Cecilia Battistelli
- Abstract
Supplementary figures S1-S8 and Supplementary Tables S1 and S2. Supplementary Figure S1: Structure conservation between human and mouse HOTAIR. Supplementary Figure S2: HOTAIR-sbid retains a nuclear localization. Supplementary Figure S3: Hep3B grown at low density express HOTAIR and Snail and repress epithelial markers. Supplementary Figure S4: Densitometric analysis of protein levels. Supplementary Figure S5: HOTAIR-sbid interferes with the SNAIL/HOTAIR/EZH2-mediated functions. Supplementary Figure S6: Quantification of migration and invasion abilities. Supplementary Figure S7: Proliferation assays. Supplementary Figure S8: HOTAIR-sbid interferes with the SNAIL/HOTAIR/EZH2-mediated functions in EMT. Table S1: List of primers, sequence, primer pairs efficiency and r^2 values. Table S2: List of primers, sequence, primer pairs efficiency and r^2 values.
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- 2023
4. Extracellular signal-Regulated Kinase 5 (ERK5) is required for the Yes-associated protein (YAP) co-transcriptional activity
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Francesca Ippolito, Veronica Consalvi, Valeria Noce, Cecilia Battistelli, Carla Cicchini, Marco Tripodi, Laura Amicone, and Alessandra Marchetti
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Cancer Research ,Cellular and Molecular Neuroscience ,erk5 ,Immunology ,Cell Biology ,yap ,transcriptional activity - Abstract
YES-associated protein (YAP) is a transcriptional cofactor with a key role in the regulation of several cellular processes, including proliferation, differentiation and epithelial-to-mesenchymal transition (EMT), by integrating multiple cell autonomous and microenvironmental cues. YAP overexpression was found in different types of human cancer where it is associated with the acquisition of stemness properties, chemoresistance, increased cell proliferation and survival, metastasis and, ultimately, poor prognosis. YAP is the main downstream effector of the Hippo pathway, a tumor suppressive signaling able to transduce several extracellular signals. The Hippo pathway acts restraining YAP activity, since its activation induces YAP phosphorylation and cytoplasmic sequestration. However, recent observations indicate that YAP activity can be also modulated by Hippo independent/integrating pathways, still largely unexplored. In this study, we demonstrated the role of the extracellular signal-regulated kinase 5 (ERK5)/mitogen-activated protein kinase in the regulation of YAP activity. By means of ERK5 inhibition/silencing and overexpression experiments, and by using as model liver stem cells, hepatocytes and hepatocellular carcinoma (HCC) cell lines, we provided evidence that ERK5 is required for YAP-dependent gene expression. Mechanistically, ERK5 controls the recruitment of YAP on promoters of target genes and its physical interaction with the transcriptional partner TEAD; moreover, it mediates the YAP activation occurring in cell adhesion, migration and TGFβ-induced EMT of liver cells. Furthermore, we demonstrated that ERK5 signaling modulates YAP activity in a LATS1/2-independent manner. Therefore, our observations identify ERK5 as a novel upstream Hippo-independent regulator of YAP activity, thus unveiling a new target for therapeutic approaches aimed at interfering with its function.
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- 2023
5. Restoration of WT1/miR-769-5p axis by HDAC1 inhibition promotes MMT reversal in mesenchymal-like mesothelial cells
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Giulio Bontempi, Michela Terri, Sabrina Garbo, Claudia Montaldo, Davide Mariotti, Veronica Bordoni, Sergio Valente, Clemens Zwergel, Antonello Mai, Alessandra Marchetti, Alessandro Domenici, Paolo Menè, Cecilia Battistelli, Marco Tripodi, and Raffaele Strippoli
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MicroRNAs ,Cancer Research ,Cellular and Molecular Neuroscience ,Epithelial-Mesenchymal Transition ,Cell Movement ,Immunology ,Cell Biology ,Epithelium - Abstract
Histone acetylation/deacetylation play an essential role in modifying chromatin structure and in regulating cell plasticity in eukaryotic cells. Therefore, histone deacetylase (HDAC) pharmacological inhibitors are promising tools in the therapy of fibrotic diseases and in cancer. Peritoneal fibrosis is a pathological process characterized by many cellular and molecular alterations, including the acquisition of invasive/pro-fibrotic abilities by mesothelial cells (MCs) through induction of mesothelial to mesenchymal transition (MMT). The aim of this study was to characterize the molecular mechanism of the antifibrotic role of HDAC1 inhibition. Specifically, treatment with MS-275, an HDAC1-3 inhibitor previously known to promote MMT reversal, induced the expression of several TGFBRI mRNA-targeting miRNAs. Among them, miR-769-5p ectopic expression was sufficient to promote MMT reversal and to limit MC migration and invasion, whereas miR-769-5p silencing further enhanced mesenchymal gene expression. These results were confirmed by HDAC1 genetic silencing. Interestingly, miR-769-5p silencing maintained mesenchymal features despite HDAC1 inhibition, thus indicating that it is necessary to drive MMT reversal induced by HDAC1 inhibition. Besides TGFBRI, miR-769-5p was demonstrated to target SMAD2/3 and PAI-1 expression directly. When analyzing molecular mechanisms underlying miR-769-5p expression, we found that the transcription factor Wilms’ tumor 1 (WT1), a master gene controlling MC development, binds to the miR-769-5p promoter favoring its expression. Interestingly, both WT1 expression and binding to miR-769-5p promoter were increased by HDAC1 inhibition and attenuated by TGFβ1 treatment. Finally, we explored the significance of these observations in the cell-to-cell communication: we evaluated the ability of miR-769-5p to be loaded into extracellular vesicles (EVs) and to promote MMT reversal in recipient mesenchymal-like MCs. Treatment of fibrotic MCs with EVs isolated from miR-769-5p over-expressing MCs promoted the down-regulation of specific mesenchymal targets and the reacquisition of an epithelial-like morphology. In conclusion, we highlighted an HDAC1-WT1-miR-769-5p axis potentially relevant for therapies aimed at counteracting organ fibrosis.
- Published
- 2022
6. Correction: Identification of a novel quinoline-based DNA demethylating compound highly potent in cancer cells
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Clemens Zwergel, Michael Schnekenburger, Federica Sarno, Cecilia Battistelli, Maria Cristina Manara, Giulia Stazi, Roberta Mazzone, Rossella Fioravanti, Christina Gros, Frédéric Ausseil, Cristina Florean, Angela Nebbioso, Raffaele Strippoli, Toshikazu Ushijima, Katia Scotlandi, Marco Tripodi, Paola B. Arimondo, Lucia Altucci, Marc Diederich, Antonello Mai, and Sergio Valente
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Genetics ,Molecular Biology ,Genetics (clinical) ,Developmental Biology - Published
- 2022
7. Relaxin/insulin-like family peptide receptor 4 (Rxfp4) expressing hypothalamic neurons modulate food intake and preference in mice
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Jo E. Lewis, Orla RM. Woodward, Danaé Nuzzaci, Christopher A. Smith, Alice E. Adriaenssens, Lawrence Billing, Cheryl Brighton, Benjamin U. Phillips, John A. Tadross, Sarah J. Kinston, Ernesto Ciabatti, Berthold Göttgens, Marco Tripodi, David Hornigold, David Baker, Fiona M. Gribble, Frank Reimann, Reimann, Frank [0000-0001-9399-6377], and Apollo - University of Cambridge Repository
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Neurons ,RXFP4 ,Mice ,Eating ,Food intake ,Hypothalamus ,Insl5 ,Animals ,Cell Biology ,CNS ,VMH ,Molecular Biology ,Receptors, G-Protein-Coupled - Abstract
OBJECTIVE: Insulin-like peptide 5 (INSL5) signalling, through its cognate receptor relaxin/insulin-like family peptide receptor 4 (RXFP4), has been reported to be orexigenic, and the high fat diet (HFD) preference observed in wildtype mice is altered in Rxfp4 knock-out mice. In this study, we used a new Rxfp4-Cre mouse model to investigate the mechanisms underlying these observations. METHODS: We generated transgenic Rxfp4-Cre mice and investigated central expression of Rxfp4 by RT-qPCR, RNAscope and intraparenchymal infusion of INSL5. Rxfp4-expressing cells were chemogenetically manipulated in global Cre-reporter mice using designer receptors exclusively activated by designer drugs (DREADDs) or after stereotactic injection of a Cre-dependent AAV-DIO-Dq-DREADD targeting a population located in the ventromedial hypothalamus (RXFP4VMH). Food intake and feeding motivation were assessed in the presence and absence of a DREADD agonist. Rxfp4-expressing cells in the hypothalamus were characterised by single-cell RNA-sequencing (scRNAseq) and the connectivity of RXFP4VMH cells was investigated using viral tracing. RESULTS: Rxfp4-Cre mice displayed Cre-reporter expression in the hypothalamus. Active expression of Rxfp4 in the adult mouse brain was confirmed by RT-qPCR and RNAscope. Functional receptor expression was supported by cyclic AMP-responses to INSL5 application in ex vivo brain slices and increased HFD and highly palatable liquid meal (HPM), but not chow, intake after intra-VMH INSL5 infusion. scRNAseq of hypothalamic RXFP4 neurons defined a cluster expressing VMH markers, alongside known appetite-modulating neuropeptide receptors (Mc4r, Cckar and Nmur2). Viral tracing demonstrated RXFP4VMH neural projections to nuclei implicated in hedonic feeding behaviour. Whole body chemogenetic inhibition (Di-DREADD) of Rxfp4-expressing cells, mimicking physiological INSL5-RXFP4 Gi-signalling, increased intake of the HFD and HPM, but not chow, whilst activation (Dq-DREADD), either at whole body level or specifically within the VMH, reduced HFD and HPM intake and motivation to work for the HPM. CONCLUSION: These findings identify RXFP4VMH neurons as regulators of food intake and preference, and hypothalamic RXFP4 signalling as a target for feeding behaviour manipulation.
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- 2022
8. Next RNA Therapeutics: The Mine of Non-Coding
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Sabrina Garbo, Rossella Maione, Marco Tripodi, and Cecilia Battistelli
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Inorganic Chemistry ,MicroRNAs ,RNA, Untranslated ,Gene Expression Regulation ,Organic Chemistry ,General Medicine ,Physical and Theoretical Chemistry ,Molecular Biology ,Spectroscopy ,Catalysis ,Computer Science Applications - Abstract
The growing knowledge on several classes of non-coding RNAs (ncRNAs) and their different functional roles has aroused great interest in the scientific community. Beyond the Central Dogma of Biology, it is clearly known that not all RNAs code for protein products, and they exert a broader repertoire of biological functions. As described in this review, ncRNAs participate in gene expression regulation both at transcriptional and post-transcriptional levels and represent critical elements driving and controlling pathophysiological processes in multicellular organisms. For this reason, in recent years, a great boost was given to ncRNA-based strategies with potential therapeutic abilities, and nowadays, the use of RNA molecules is experimentally validated and actually exploited in clinics to counteract several diseases. In this review, we summarize the principal classes of therapeutic ncRNA molecules that are potentially implied in disease onset and progression, which are already used in clinics or under clinical trials, highlighting the advantages and the need for a targeted therapeutic strategy design. Furthermore, we discuss the benefits and the limits of RNA therapeutics and the ongoing development of delivery strategies to limit the off-target effects and to increase the translational application.
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- 2022
9. SYNCRIP modulates the epithelial-mesenchymal transition in hepatocytes and HCC Cells
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Veronica Riccioni, Flavia Trionfetti, Claudia Montaldo, Sabrina Garbo, Francesco Marocco, Cecilia Battistelli, Alessandra Marchetti, Raffaele Strippoli, Laura Amicone, Carla Cicchini, and Marco Tripodi
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Carcinoma, Hepatocellular ,Epithelial-Mesenchymal Transition ,QH301-705.5 ,HnRNPQ ,hnRNPQ ,EMT ,mirRNAs ,HCC ,RNA binding proteins ,metastasis ,Metastasis ,MirRNAs ,Article ,Heterogeneous-Nuclear Ribonucleoproteins ,Catalysis ,Inorganic Chemistry ,Mice ,Cell Movement ,Cell Line, Tumor ,Animals ,Physical and Theoretical Chemistry ,Biology (General) ,Molecular Biology ,QD1-999 ,Spectroscopy ,Liver Neoplasms ,Organic Chemistry ,RNA-Binding Proteins ,General Medicine ,Computer Science Applications ,Gene Expression Regulation, Neoplastic ,MicroRNAs ,Chemistry ,Cell Transformation, Neoplastic ,Phenotype ,Gene Knockdown Techniques ,Hepatocytes ,RNA Interference ,Disease Susceptibility ,Biomarkers - Abstract
Heterogeneous nuclear ribonucleoproteins (hnRNPs) control gene expression by acting at multiple levels and are often deregulated in epithelial tumors; however, their roles in the fine regulation of cellular reprogramming, specifically in epithelial–mesenchymal transition (EMT), remain largely unknown. Here, we focused on the hnRNP-Q (also known as SYNCRIP), showing by molecular analysis that in hepatocytes it acts as a “mesenchymal” gene, being induced by TGFβ and modulating the EMT. SYNCRIP silencing limits the induction of the mesenchymal program and maintains the epithelial phenotype. Notably, in HCC invasive cells, SYNCRIP knockdown induces a mesenchymal–epithelial transition (MET), negatively regulating their mesenchymal phenotype and significantly impairing their migratory capacity. In exploring possible molecular mechanisms underlying these observations, we identified a set of miRNAs (i.e., miR-181-a1-3p, miR-181-b1-3p, miR-122-5p, miR-200a-5p, and miR-let7g-5p), previously shown to exert pro- or anti-EMT activities, significantly impacted by SYNCRIP interference during EMT/MET dynamics and gathered insights, suggesting the possible involvement of this RNA binding protein in their transcriptional regulation.
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- 2022
10. Pleural Mesothelial Cells Modulate the Inflammatory/Profibrotic Response During SARS-CoV-2 Infection
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Giulia Matusali, Flavia Trionfetti, Veronica Bordoni, Roberta Nardacci, Laura Falasca, Daniele Colombo, Michela Terri, Claudia Montaldo, Concetta Castilletti, Davide Mariotti, Franca Del Nonno, Maria Rosaria Capobianchi, Chiara Agrati, Marco Tripodi, and Raffaele Strippoli
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mesothelial cells ,pulmonary fibrosis ,QH301-705.5 ,SARS-CoV-2 ,inflammatory cytokines ,business.industry ,mesothelial to mesenchymal transition ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,respiratory system ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,Biochemistry ,WT1 ,respiratory tract diseases ,Immunology ,Medicine ,Molecular Biosciences ,Biology (General) ,business ,Molecular Biology ,Mesothelial Cell ,Original Research - Abstract
Although lung fibrosis has a major impact in COVID-19 disease, its pathogenesis is incompletely understood. In particular, no direct evidence of pleura implication in COVID-19-related fibrotic damage has been reported so far. In this study, the expression of epithelial cytokeratins and Wilms tumor 1 (WT1), specific markers of mesothelial cells (MCs), was analyzed in COVID-19 and unrelated pleura autoptic samples. SARS-CoV-2 replication was analyzed by RT-PCR and confocal microscopy in MeT5A, a pleura MC line. SARS-CoV-2 receptors were analyzed by RT-PCR and western blot. Inflammatory cytokines from the supernatants of SARS-CoV-2-infected MeT5A cells were analysed by Luminex and ELLA assays. Immunohistochemistry of COVID-19 pleura patients highlighted disruption of pleura monolayer and fibrosis of the sub-mesothelial stroma, with the presence of MCs with fibroblastoid morphology in the sub-mesothelial stroma, but no evidence of direct infection in vivo. Interestingly, we found evidence of ACE2 expression in MCs from pleura of COVID-19 patients. In vitro analysis shown that MeT5A cells expressed ACE2, TMPRSS2, ADAM17 and NRP1, plasma membrane receptors implicated in SARS-CoV-2 cell entry and infectivity. Moreover, MeT5A cells sustained SARS-CoV-2 replication and productive infection. Infected MeT5A cells produced interferons, inflammatory cytokines and metalloproteases. Overall, our data highlight the potential role of pleura MCs as promoters of the fibrotic reaction and regulators of the immune response upon SARS-CoV-2 infection.
- Published
- 2021
11. 15 Eni’s projects in Italy for hydrogen production
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Giuseppe Ricci, Maurizio Dessì, Marco Tripodi, Paolo Fiaschi, Roberto Palmieri, Luca Eugenio Basini, Thomas Pasini, and Alessandra Guarinoni
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- 2021
12. 14 Hydrogen applications in ENI: from Industrial applications to power generation
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Maurizio Dessi, Laura Prosperi, Marco Tripodi, and Giuseppe Ricci
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Electricity generation ,Materials science ,Hydrogen ,chemistry ,business.industry ,chemistry.chemical_element ,Process engineering ,business - Published
- 2021
13. Relaxin/insulin-like family peptide receptor 4 (Rxfp4) expressing hypothalamic neurons modulate food intake and preference in mice
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Lewis Je, Lawrence J. Billing, Bertie Gottgens, Fiona M. Gribble, Christopher A. Smith, John Tadross, Kinston Sj, Cheryl A. Brighton, David Baker, Marco Tripodi, Hornigold D, Woodward Or, Benjamin U. Phillips, Ciabatti E, Alice E. Adriaenssens, and Frank Reimann
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Relaxin ,medicine.medical_specialty ,Central nucleus of the amygdala ,Biology ,Oxytocin receptor ,Stria terminalis ,Endocrinology ,medicine.anatomical_structure ,Hypothalamus ,Internal medicine ,medicine ,Receptor ,Relaxin/insulin-like family peptide receptor 4 ,Nucleus - Abstract
Relaxin/insulin-like-family peptide receptor-4 (RXFP4), the cognate receptor for insulin-like peptide 5 (INSL5), has been implicated in feeding behaviour as Rxfp4 knockout mice display shorter meal durations and reduced high fat diet (HFD) intake. Here, we generated transgenic Rxfp4-Cre mice to explore Rxfp4 expression and physiology. Using this model, we identified Rxfp4 expression in the central nervous system, including in the ventromedial hypothalamus (VMH). Intra-VMH infusion of INSL5 increased HFD and highly palatable liquid meal intake (HPM) of ad libitum fed wildtype mice. Single-cell RNA-sequencing of VMH Rxfp4-expressing cells (RXFP4VMH) defined a cluster of Rxfp4-labelled neurons expressing Esr1, Tac1 and Oxtr, alongside known appetite-modulating neuropeptide receptors (Mc4r, Cckar and Nmur2). Viral tracing demonstrated RXFP4VMH neural projections to the bed nucleus of the stria terminalis, paraventricular hypothalamus, paraventricular thalamus and central nucleus of the amygdala. Utilising designer receptors exclusively activated by designer drugs (DREADDs), we found that whole body chemogenetic inhibition (Di) of Rxfp4-expressing cells, mimicking native INSL5-RXFP4 signalling, increased intake of HFD and HPM, whilst activation (Dq), either at whole body level or specifically within the VMH, reduced HFD and HPM intake and altered food preference. Ablating VMH Rxfp4-expressing cells recapitulated the lower HFD intake phenotype of Rxfp4 knockout mice, resulting in reduced body weight. These findings identify a discrete Rxfp4-expressing neuronal population as a key regulator of food intake and preference and reveal hypothalamic RXFP4 signalling as a target for feeding behaviour manipulation.
- Published
- 2021
14. Goal-Oriented Behaviour: The Ventral Tegmental Area in Motivated Movements
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Marco Tripodi and Laura Masullo
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0301 basic medicine ,Goal orientation ,Dopaminergic ,Biology ,Inhibitory postsynaptic potential ,General Biochemistry, Genetics and Molecular Biology ,Ventral tegmental area ,Midbrain ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,medicine.anatomical_structure ,nervous system ,medicine ,Head movements ,General Agricultural and Biological Sciences ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Summary The ventral tegmental area is a midbrain region known for the involvement of its dopaminergic neurons in encoding reward-related features, value and motivational states. New research suggests a role for inhibitory neurons of the ventral tegmental area in the orchestration of head movements, which might be instrumental in guiding animals towards spatial targets during motivated behaviour.
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- 2019
15. Persistence of fibrogenic process in direct-acting antiviral (DAA) treated patients?
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Cecilia Battistelli and Marco Tripodi
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Molecular Biology - Published
- 2022
16. Mechanisms of Peritoneal Fibrosis: Focus on Immune Cells–Peritoneal Stroma Interactions
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Michela Terri, Flavia Trionfetti, Claudia Montaldo, Marco Cordani, Marco Tripodi, Manuel Lopez-Cabrera, Raffaele Strippoli, European Commission, and Ministerio de Ciencia e Innovación (España)
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0301 basic medicine ,lcsh:Immunologic diseases. Allergy ,Pathology ,medicine.medical_specialty ,medicine.medical_treatment ,Immunology ,030232 urology & nephrology ,Peritonitis ,Inflammation ,pro-inflammatory cytokines ,Peritoneal dialysis ,Extracellular matrix ,peritoneal fibrosis ,mesothelial cells ,peritonitis ,innate immunity ,T cell subpopulations ,03 medical and health sciences ,0302 clinical medicine ,Fibrosis ,medicine ,Immunology and Allergy ,Mesothelial cells ,Peritoneal Fibrosis ,Innate immunity ,Proinflammatory cytokines ,Innate immune system ,Peritoneal fibrosis ,business.industry ,medicine.disease ,030104 developmental biology ,Systematic Review ,medicine.symptom ,business ,lcsh:RC581-607 ,Myofibroblast - Abstract
Peritoneal fibrosis is characterized by abnormal production of extracellular matrix proteins leading to progressive thickening of the submesothelial compact zone of the peritoneal membrane. This process may be caused by a number of insults including pathological conditions linked to clinical practice, such as peritoneal dialysis, abdominal surgery, hemoperitoneum, and infectious peritonitis. All these events may cause acute/chronic inflammation and injury to the peritoneal membrane, which undergoes progressive fibrosis, angiogenesis, and vasculopathy. Among the cellular processes implicated in these peritoneal alterations is the generation of myofibroblasts from mesothelial cells and other cellular sources that are central in the induction of fibrosis and in the subsequent functional deterioration of the peritoneal membrane. Myofibroblast generation and activity is actually integrated in a complex network of extracellular signals generated by the various cellular types, including leukocytes, stably residing or recirculating along the peritoneal membrane. Here, the main extracellular factors and the cellular players are described with emphasis on the cross-talk between immune system and cells of the peritoneal stroma. The understanding of cellular and molecular mechanisms underlying fibrosis of the peritoneal membrane has both a basic and a translational relevance, since it may be useful for setup of therapies aimed at counteracting the deterioration as well as restoring the homeostasis of the peritoneal membrane., European Union’s Horizon 2020 Research and Innovation Programme under the Marie Sklodowska-Curie grant and Spanish Ministry of Science and Innovation/Fondo Europeo de Desarrollo Regional (PID2019-110132RB-I00/AEI/10.13039/501100011033)
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- 2021
17. The RNA editing enzyme ADAR2 restricts L1 mobility
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Silvia Galardi, Sofian Alwardat, Loredana Frassinelli, Alessandro Michienzi, Marco Tripodi, Elisa Orecchini, Margherita Doria, Silvia Anna Ciafrè, and Carmine Mancone
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RNA editing ,Adenosine Deaminase ,Retrotransposon ,Biology ,LINE-1 ,medicine ,ADAR2 ,Humans ,Molecular Biology ,chemistry.chemical_classification ,Settore BIO/13 ,double-stranded RNA (dsRNA) ,retrotransposons ,RNA-Binding Proteins ,RNA ,Cell Biology ,Adenosine ,HEK293 Cells ,Long Interspersed Nucleotide Elements ,Enzyme ,Biochemistry ,chemistry ,HeLa Cells ,Research Article ,Research Paper ,medicine.drug - Abstract
Adenosine deaminases acting on RNA (ADARs) are enzymes that convert adenosines to inosines in double-stranded RNAs (RNA editing A-to-I). ADAR1 and ADAR2 were previously reported as HIV-1 proviral factors. The aim of this study was to investigate the composition of the ADAR2 ribonucleoprotein complex during HIV-1 expression. By using a dual-tag affinity purification procedure in cells expressing HIV-1 followed by mass spectrometry analysis, we identified 10 non-ribosomal ADAR2-interacting factors. A significant fraction of these proteins was previously found associated to the Long INterspersed Element 1 (LINE1 or L1) ribonucleoparticles and to regulate the life cycle of L1 retrotransposons. Considering that we previously demonstrated that ADAR1 is an inhibitor of LINE-1 retrotransposon activity, we investigated whether also ADAR2 played a similar function. To reach this goal, we performed specific cell culture retrotransposition assays in cells overexpressing or ablated for ADAR2. These experiments unveil a novel function of ADAR2 as suppressor of L1 retrotransposition. Furthermore, we showed that ADAR2 binds the basal L1 RNP complex. Overall, these data support the role of ADAR2 as regulator of L1 life cycle.
- Published
- 2021
18. Fibrogenic signals persist in DAA-treated HCV patients after sustained virological response
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Michela Terri, Veronica Bordoni, Tiziana Vescovo, Claudia Montaldo, Eleonora Tartaglia, Marco Tripodi, Flavia Trionfetti, Raffaele Strippoli, Veronica Riccioni, Chiara Agrati, Maria Giulia Prado, Cecilia Battistelli, and Gianpiero D'Offizi
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Cirrhosis ,liver fibrosis ,extracellular vesicles ,direct-acting antiviral (DAA) therapy ,Hepatology ,medicine.diagnostic_test ,biology ,Sustained Virologic Response ,business.industry ,Extracellular vesicle ,Cell Communication ,Hepacivirus ,medicine.disease ,Antiviral Agents ,Hepatitis C ,Virus ,Mass Spectrometry ,Liver disease ,Western blot ,Fibrosis ,microRNA ,Immunology ,medicine ,biology.protein ,Humans ,ACTA2 ,business - Abstract
Patients with HCV who achieve a sustained virological response (SVR) on direct-acting antiviral (DAA) therapy still need to be monitored for signs of liver disease progression. To this end, the identification of both disease biomarkers and therapeutic targets is necessary.Extracellular vesicles (EVs) purified from plasma of 15 healthy donors (HDs), and 16 HCV-infected patients before (T0) and after (T6) DAA treatment were utilized for functional and miRNA cargo analysis. EVs purified from plasma of 17 HDs and 23 HCV-infected patients (T0 and T6) were employed for proteomic and western blot analyses. Functional analysis in LX2 cells measured fibrotic markers (mRNAs and proteins) in response to EVs. Structural analysis was performed by qPCR, label-free liquid chromatography-mass spectrometry and western blot.On the basis of observations indicating functional differences (i.e. modulation of FN-1, ACTA2, Smad2/3 phosphorylation, collagen deposition) of plasma-derived EVs from HDs, T0 and T6, we performed structural analysis of EVs. We found consistent differences in terms of both miRNA and protein cargos: (i) antifibrogenic miR204-5p, miR181a-5p, miR143-3p, miR93-5p and miR122-5p were statistically underrepresented in T0 EVs compared to HD EVs, while miR204-5p and miR143-3p were statistically underrepresented in T6 EVs compared to HD EVs (p0.05); (ii) proteomic analysis highlighted, in both T0 and T6, the modulation of several proteins with respect to HDs; among them, the fibrogenic protein DIAPH1 was upregulated (LogTaken together, these results highlight structural EV modifications that are conceivably causal for long-term liver disease progression in patients with HCV despite DAA-mediated SVR.Direct-acting antivirals lead to virological cure in the majority of patients with chronic hepatitis C virus infection. However, the risk of liver disease progression or complications in patients with fibrosis and cirrhosis remains in some patients even after virological cure. Herein, we show that extracellular vesicle modifications could be linked to long-term liver disease progression in patients who have achieved virological cure; these modifications could potentially be used as biomarkers or treatment targets in such patients.
- Published
- 2020
19. A novel RNA- based approach to counteract EMT
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Marco Tripodi, Sabrina Garbo, and Cecilia Battistelli
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Cancer Research ,lncRNA ,Oncology ,Research Perspective ,EMT ,RNA ,Biology ,Cell biology - Published
- 2021
20. Genomic stability of Self-inactivating Rabies
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Hassal Lee, Daniel de Malmazet, Ernesto Ciabatti, Marco Tripodi, Ana González-Rueda, and Fabio Morgese
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Genetics ,In vivo ,Rabies virus ,TEV protease ,medicine ,Genetic regulatory element ,Biology ,medicine.disease_cause ,Genome ,Function (biology) ,Virus ,Viral vector - Abstract
Transsynaptic viral vectors provide means to gain genetic access to neurons based on synaptic connectivity and are essential tools for the dissection of neural circuit function. Among them, the retrograde monosynaptic ΔG-Rabies has been widely used in neuroscience research. A recently developed engineered version of the ΔG-Rabies, the non-toxic self-inactivating (SiR) virus, represents the first tool for open-ended genetic manipulation of neural circuits. However, the high mutational rate of the rabies virus poses a risk that mutations targeting the key genetic regulatory element in the SiR genome could emerge and revert it to a canonical ΔG-Rabies. Such revertant mutations have recently been identified in a SiR batch. To address the origin, incidence and relevance of these mutations, we investigated the genomic stability of SiR in vitro and in vivo. We found that “revertant” mutations are rare and accumulate only when SiR is extensively amplified in vitro, particularly in suboptimal production cell lines that have insufficient levels of TEV protease activity. Moreover, we confirmed that SiR-CRE, unlike canonical ΔG-Rab-CRE or revertant-SiR-CRE, is non-toxic and that revertant mutations do not emerge in vivo during long-term experiments.HighlightsRevertant mutations are rare and do not accumulate when SiR is produced in high-TEVp expressing production cell linesSiR is non-toxic in vivoRevertant SiR mutations do not accumulate during in vivo experiments
- Published
- 2020
21. Design and Functional Validation of a Mutant Variant of the LncRNA
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Cecilia, Battistelli, Sabrina, Garbo, Veronica, Riccioni, Claudia, Montaldo, Laura, Santangelo, Andrea, Vandelli, Raffaele, Strippoli, Gian Gaetano, Tartaglia, Marco, Tripodi, and Carla, Cicchini
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Carcinoma, Hepatocellular ,Epithelial-Mesenchymal Transition ,Liver Neoplasms ,Apoptosis ,macromolecular substances ,Article ,Gene Expression Regulation, Neoplastic ,Mice ,Cell Movement ,Mutation ,Biomarkers, Tumor ,Hepatocytes ,Animals ,Humans ,RNA, Long Noncoding ,Snail Family Transcription Factors ,Cells, Cultured ,Cell Proliferation - Abstract
HOTAIR is a lncRNA overexpressed in several epithelial cancers and strongly correlated with invasion. This lncRNA was proven a pivotal element of the epithelial-mesenchymal transition (EMT), a trans-differentiation process triggering metastasis. Snail, master inducer of EMT, requires HOTAIR to recruit EZH2 on specific epithelial target genes (i.e., HNF4α, E-cadherin and HNF1α) and cause their repression. Here we designed a HOTAIR deletion mutant form, named HOTAIR-sbid, including the putative Snail-binding domain but depleted of the EZH2 binding domain. HOTAIR-sbid acted as a dominant negative of the endogenous HOTAIR. In both murine and human tumor cells, HOTAIR-sbid impaired the ability of HOTAIR to bind Snail and, in turn, trigger H3K27me3/EZH2-mediated repression of Snail epithelial target genes. Notably, HOTAIR-sbid expression was proven to reduce cellular motility, invasiveness, anchorage-independent growth, and responsiveness to TGFβ-induced EMT. These data provide evidence on a lncRNA-based strategy to effectively impair the function of a master EMT-transcriptional factor.
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- 2020
22. Hypoxia-Induced miR-675-5p Supports β-Catenin Nuclear Localization by Regulating GSK3-β Activity in Colorectal Cancer Cell Lines
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Marco Tripodi, Alice Conigliaro, Maria Magdalena Barreca, Maria Giulia Prado, Laura Saieva, Riccardo Alessandro, Chiara Zichittella, Saieva, Laura, Barreca, Maria Magdalena, Zichittella, Chiara, Prado, Maria Giulia, Tripodi, Marco, Alessandro, Riccardo, and Conigliaro, Alice
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0301 basic medicine ,Transcription factor complex ,Kaplan-Meier Estimate ,lcsh:Chemistry ,0302 clinical medicine ,GSK-3 ,poxiahy ,lcsh:QH301-705.5 ,long non-coding H19 ,Spectroscopy ,beta Catenin ,Kinase ,Chemistry ,General Medicine ,Cell Hypoxia ,Computer Science Applications ,Cell biology ,Gene Expression Regulation, Neoplastic ,030220 oncology & carcinogenesis ,Colorectal Neoplasms ,Protein Binding ,Active Transport, Cell Nucleus ,colorectal cancer ,miR-675 ,Transfection ,Catalysis ,Article ,Inorganic Chemistry ,03 medical and health sciences ,Cell Line, Tumor ,microRNA ,Gene silencing ,Humans ,Physical and Theoretical Chemistry ,Molecular Biology ,Glycogen Synthase Kinase 3 beta ,hypoxia ,Organic Chemistry ,RNA ,Computational Biology ,β-catenin ,HCT116 Cells ,MicroRNAs ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,Microscopy, Fluorescence ,Catenin ,Mutation ,Nuclear localization sequence - Abstract
The reduction of oxygen partial pressure in growing tumors triggers numerous survival strategies driven by the transcription factor complex HIF1 (Hypoxia Inducible Factor-1). Recent evidence revealed that HIF1 promotes rapid and effective phenotypic changes through the induction of non-coding RNAs, whose contribution has not yet been fully described. Here we investigated the role of the hypoxia-induced, long non-coding RNA H19 (lncH19) and its intragenic miRNA (miR-675-5p) into HIF1-Wnt crosstalk. During hypoxic stimulation, colorectal cancer cell lines up-regulated the levels of both the lncH19 and its intragenic miR-675-5p. Loss of expression experiments revealed that miR-675-5p inhibition, in hypoxic cells, hampered &beta, catenin nuclear localization and its transcriptional activity, while lncH19 silencing did not induce the same effects. Interestingly, our data revealed that miRNA inhibition in hypoxic cells restored the activity of Glycogen Synthase Kinase 3&beta, (GSK-3&beta, ) reducing the amount of P-Ser9 kinase, thus unveiling a role of the miR-675-5p in controlling GSK-3&beta, activity. Bioinformatics analyses highlighted the serine/threonine-protein phosphatases PPP2CA, responsible for GSK-3&beta, activation, among the miR-675-5p targets, thus indicating the molecular mediator through which miR-675-5p may control &beta, catenin nuclear localization. In conclusion, here we demonstrated that the inhibition of the hypoxia-induced non-coding RNA miR-675-5p hampered the nuclear localization of &beta, catenin by regulating GSK-3&beta, activity, thus proposing the miR-675-5p as a new therapeutic target for the treatment of colorectal cancer.
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- 2020
23. Design of First-in-Class Dual {EZH}2/{HDAC} Inhibitor: biochemical activity and biological evaluation in cancer cells
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Annalisa Romanelli, Clemens Zwergel, Rossella Fioravanti, Sergio Valente, Raffaele Strippoli, Rossella Rota, Marco Tripodi, Elisabetta Di Bello, Daniela Trisciuoglio, Giulia Stazi, Silvia Pomella, Clara Perrone, Donatella Del Bufalo, Cecilia Battistelli, and Antonello Mai
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Chemistry ,medicine.drug_class ,Drug discovery ,Cellular differentiation ,histone methyltransferase ,Organic Chemistry ,Histone deacetylase inhibitor ,EZH2 ,drug discovery ,histone deacetylase ,dual-target inhibitor ,anticancer agent ,macromolecular substances ,Biochemistry ,Settore MED/05 ,Histone methyltransferase ,Cancer cell ,medicine ,Cancer research ,Epithelial–mesenchymal transition ,Viability assay ,Histone deacetylase - Abstract
[Image: see text] Since the histone modifying enzymes EZH2 and HDACs control a number of epigenetic-dependent carcinogenic pathways, we designed the first-in-class dual EZH2/HDAC inhibitor 5 displaying (sub)micromolar inhibition against both targets. When tested in several cancer cell lines, the hybrid 5 impaired cell viability at low micromolar level and in leukemia U937 and rhabdomyosarcoma RH4 cells provided G1 arrest, apoptotic induction, and increased differentiation, associated with an increase of acetyl-H3 and acetyl-α-tubulin and a decrease of H3K27me3 levels. In glioblastoma U87 cells, 5 hampered epithelial to mesenchymal transition by increasing the E-cadherin expression, thus proposing itself as a useful candidate for anticancer therapy.
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- 2020
24. Novel quinoline compounds active in cancer cells through coupled DNA methyltransferase inhibition and degradation
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Clemens Zwergel, Marco Tripodi, Donatella Del Bufalo, Annalisa Romanelli, Daniela Trisciuoglio, Cecilia Battistelli, Lucia Altucci, Giulia Stazi, Paola B. Arimondo, Rossella Fioravanti, Teresa De Luca, Alexandra Paulo, Dany Pechalrieu, Sergio Valente, Raffaele Strippoli, Angela Nebbioso, Federica Sarno, Eduarda Mendes, Antonello Mai, Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome] (UNIROMA), Università degli studi della Campania 'Luigi Vanvitelli' = University of the Study of Campania Luigi Vanvitelli, Universidade de Lisboa = University of Lisbon (ULISBOA), Istituto Nazionale di Malattie Infettive 'Lazzaro Spallanzani' (INMI), Department of Molecular Medicine, Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome] (UNIROMA), Pharmacochimie de la Régulation Epigénétique du Cancer (ETaC), PIERRE FABRE-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Chimie biologique épigénétique - Epigenetic Chemical Biology (EpiCBio), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), IFO - Istituto Nazionale Tumori Regina Elena [Roma] (IRE), CNR Istituto di Biologia e Patologia Molecolari [Roma] (CNR | IBPM), National Research Council of Italy | Consiglio Nazionale delle Ricerche (CNR), This work was supported by PRIN 2016 (prot. 20152TE5PK) (L.A., A.M.), Ricerca Finalizzata 2013 PE-2013-02355271 (A.M.), the Italian Association for Cancer Research AIRC-19162 (A.M.), AIRC-17217 (L.A.) and AIRC-18560 (D.D.B.), NIH (n. R01GM114306) (A.M.), VALERE: Vanvitelli per la Ricerca Program (L.A.), FP7-BLUEPRINT (282510) (L.A., A.M.), the Campania Regional Government Lotta alle Patologie Oncologiche (L.A.), iCURE (CUP B21c17000030007) (L.A.), Campania Regional Government FASE 2: IDEAL (CUP B63D18000560007) (L.A.), PlanCancer2014 (P.B.A.) and FCT, Portugal through UID/DTP/04138/2019 (A.P., E.M.) funds., Zwergel, C., Fioravanti, R., Stazi, G., Sarno, F., Battistelli, C., Romanelli, A., Nebbioso, A., Mendes, E., Paulo, A., Strippoli, R., Tripodi, M., Pechalrieu, D., Arimondo, P. B., De Luca, T., Del Bufalo, D., Trisciuoglio, D., Altucci, L., Valente, S., Mai, A., Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], Università degli studi della Campania 'Luigi Vanvitelli', Universidade de Lisboa (ULISBOA), Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome]-Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institute of Molecular Pathology and Biology [Rome] (IPBM), and Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome]-Consiglio Nazionale delle Ricerche (CNR)
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0301 basic medicine ,Cancer Research ,Methyltransferase ,[SDV]Life Sciences [q-bio] ,Azacitidine ,Decitabine ,DNA methyltransferase ,Apoptosis ,Medicinal chemistry ,Protein degradation ,lcsh:RC254-282 ,Article ,03 medical and health sciences ,0302 clinical medicine ,medicine ,[CHIM]Chemical Sciences ,Chemistry ,Drug discovery ,Apoptosi ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,3. Good health ,Enzyme inhibition ,030104 developmental biology ,Oncology ,Cell culture ,030220 oncology & carcinogenesis ,Cancer cell ,Cancer research ,DNMT1 ,medicine.drug - Abstract
DNA methyltransferases (DNMTs) play a relevant role in epigenetic control of cancer cell survival and proliferation. Since only two DNMT inhibitors (azacitidine and decitabine) have been approved to date for the treatment of hematological malignancies, the development of novel potent and specific inhibitors is urgent. Here we describe the design, synthesis, and biological evaluation of a new series of compounds acting at the same time as DNMTs (mainly DNMT3A) inhibitors and degraders. Tested against leukemic and solid cancer cell lines, 2a&ndash, c and 4a&ndash, c (the last only for leukemias) displayed up to submicromolar antiproliferative activities. In HCT116 cells, such compounds induced EGFP gene expression in a promoter demethylation assay, confirming their demethylating activity in cells. In the same cell line, 2b and 4c chosen as representative samples induced DNMT1 and -3A protein degradation, suggesting for these compounds a double mechanism of DNMT3A inhibition and DNMT protein degradation.
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- 2020
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25. Caveolin1 and YAP drive mechanically induced mesothelial to mesenchymal transition and fibrosis
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Manuel Gómez, Fátima Sánchez-Cabo, Miguel A. del Pozo, Marta Loureiro, José A. Jiménez-Heffernan, Michela Terri, Raffaele Strippoli, Lucía Pascual-Antón, Pilar Sandoval, Manuel López-Cabrera, Cecilia Battistelli, Jesús Vázquez, Marco Tripodi, Víctor Jiménez-Jiménez, Francesca Matteini, Enrique Calvo, Roberto Moreno-Vicente, Lucia Rossi, Ministerio de Economía y Competitividad (España), Ministerio de Ciencia e Innovación (España), Fundació La Marató, Worldwide Cancer Research, European Regional Development Fund (ERDF/FEDER), Sapineza University of Rome, Instituto de Salud Carlos III - ISCIII, Fundación ProCNIC, Fundació La Marató de TV3, Sapienza Università di Roma, Centro Nacional de Investigaciones Cardiovasculares (España), Instituto de Salud Carlos III, Fundación La Marató TV3, Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF), and Sapienza University of Rome (Italia)
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Male ,Cancer Research ,Epithelial-Mesenchymal Transition ,Caveolin 1 ,Immunology ,Tissue Adhesions ,Caveolins ,Article ,Transforming Growth Factor beta1 ,Transcriptome ,Mice ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,Downregulation and upregulation ,Fibrosis ,medicine ,Animals ,Humans ,Smad3 Protein ,lcsh:QH573-671 ,Mechanotransduction ,Peritoneal Fibrosis ,Adaptor Proteins, Signal Transducing ,030304 developmental biology ,0303 health sciences ,lcsh:Cytology ,Chemistry ,Mesenchymal stem cell ,Epithelial Cells ,YAP-Signaling Proteins ,Chronic inflammation ,Cell Biology ,medicine.disease ,fibrosis mesothelial cells mechanotransduction YAP Caveolin1 ,3. Good health ,Cell biology ,Mice, Inbred C57BL ,Disease Models, Animal ,Crosstalk (biology) ,Mechanisms of disease ,030220 oncology & carcinogenesis ,Female ,Peritoneum ,Signal transduction ,Peritoneal Dialysis ,Signal Transduction ,Transcription Factors - Abstract
Despite their emerging relevance to fully understand disease pathogenesis, we have as yet a poor understanding as to how biomechanical signals are integrated with specific biochemical pathways to determine cell behaviour. Mesothelial-to-mesenchymal transition (MMT) markers colocalized with TGF-β1-dependent signaling and yes-associated protein (YAP) activation across biopsies from different pathologies exhibiting peritoneal fibrosis, supporting mechanotransduction as a central driving component of these class of fibrotic lesions and its crosstalk with specific signaling pathways. Transcriptome and proteome profiling of the response of mesothelial cells (MCs) to linear cyclic stretch revealed molecular changes compatible with bona fide MMT, which (i) overlapped with established YAP target gene subsets, and were largely dependent on endogenous TGF-β1 signaling. Importantly, TGF-β1 blockade blunts the transcriptional upregulation of these gene signatures, but not the mechanical activation and nuclear translocation of YAP per se. We studied the role therein of caveolin-1 (CAV1), a plasma membrane mechanotransducer. Exposure of CAV1-deficient MCs to cyclic stretch led to a robust upregulation of MMT-related gene programs, which was blunted upon TGF-β1 inhibition. Conversely, CAV1 depletion enhanced both TGF-β1 and TGFBRI expression, whereas its re-expression blunted mechanical stretching-induced MMT. CAV1 genetic deficiency exacerbated MMT and adhesion formation in an experimental murine model of peritoneal ischaemic buttons. Taken together, these results support that CAV1-YAP/TAZ fine-tune the fibrotic response through the modulation of MMT, onto which TGF-β1-dependent signaling coordinately converges. Our findings reveal a cooperation between biomechanical and biochemical signals in the triggering of MMT, representing a novel potential opportunity to intervene mechanically induced disorders coursing with peritoneal fibrosis, such as post-surgical adhesions., MICINN and MINECO (SAF2014-51876-R and SAF2017-83130-R; MINSEV1512-07-2016; CSD2009-0016 and BFU2016-81912-REDC), Fundació La Marató de TV3 (674/C/2013); Worldwide Cancer Research; Sapienza University of Rome (RM116154BE5E14B2); CNIC is supported by the ISCIII, the MCIN and the Pro CNIC Foundation, and is a Severo Ochoa Center of Excellence (SEV-2015-0505
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- 2020
26. The lncRNA HOTAIR transcription is controlled by HNF4α-induced chromatin topology modulation
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Laura Santangelo, Marco Tripodi, Cecilia Battistelli, Frank J. Gonzalez, Carla Cicchini, Giovanna Sabarese, and Claudia Montaldo
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0301 basic medicine ,Carcinoma, Hepatocellular ,Epithelial-Mesenchymal Transition ,Biology ,Article ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Transcription (biology) ,snail ,Transcriptional regulation ,Animals ,Humans ,Hox gene ,Molecular Biology ,epigenetics ,Liver Neoplasms ,EMT ,RNA ,Cell Differentiation ,HOTAIR ,Oncogenes ,Cell Biology ,Chromatin ,LncRNA ,Long non-coding RNA ,Cell biology ,Gene Expression Regulation, Neoplastic ,Hepatocyte nuclear factors ,030104 developmental biology ,Gene Expression Regulation ,Hepatocyte Nuclear Factor 4 ,030220 oncology & carcinogenesis ,Hepatocytes ,RNA, Long Noncoding ,Snail Family Transcription Factors - Abstract
The expression of the long noncoding RNA HOTAIR (HOX Transcript Antisense Intergenic RNA) is largely deregulated in epithelial cancers and positively correlates with poor prognosis and progression of hepatocellular carcinoma and gastrointestinal cancers. Furthermore, functional studies revealed a pivotal role for HOTAIR in the epithelial-to-mesenchymal transition, as this RNA is causal for the repressive activity of the master factor SNAIL on epithelial genes. Despite the proven oncogenic role of HOTAIR, its transcriptional regulation is still poorly understood. Here hepatocyte nuclear factor 4-α (HNF4α), as inducer of epithelial differentiation, was demonstrated to directly repress HOTAIR transcription in the mesenchymal-to epithelial transition. Mechanistically, HNF4α was found to cause the release of a chromatin loop on HOTAIR regulatory elements thus exerting an enhancer-blocking activity.
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- 2018
27. Looking at Synaptic Specificity from a Different Angle
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Marco Tripodi and Letizia Mariotti
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Neurons ,0303 health sciences ,General Neuroscience ,030302 biochemistry & molecular biology ,Motor Cortex ,Sensation ,Biology ,Article ,03 medical and health sciences ,medicine.anatomical_structure ,nervous system ,Synaptic specificity ,medicine ,Neuron ,Neuroscience ,030304 developmental biology - Abstract
Proprioceptive sensory axons in the spinal cord form selective connections with motor neuron partners, but the strategies that confer such selectivity remain uncertain. We show that muscle-specific sensory axons project to motor neurons along topographically organized angular trajectories and that motor pools exhibit diverse dendritic arbors. On the basis of spatial constraints on axo-dendritic interactions, we propose positional strategies that can account for sensory-motor connectivity and synaptic organization. These strategies rely on two patterning principles. First, the degree of axo-dendritic overlap reduces the number of potential post-synaptic partners. Second, a close correlation between the small angle of axo-dendritic approach and the formation of synaptic clusters imposes specificity of connections when sensory axons intersect multiple motor pools with overlapping dendritic arbors. Our study identifies positional strategies with prominent roles in the organization of spinal sensory-motor circuits.
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- 2019
28. Nonenzymatic Oligomerization of 3′,5′-Cyclic CMP Induced by Proton and UV Irradiation Hints at a Nonfastidious Origin of RNA
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Carmine Mancone, Samanta Pino, Michal Otyepka, Anita Scipioni, Ernesto Di Mauro, Holger Kruse, Alessandra Giorgi, Eugene Krasavin, Jiří Šponer, Marco Tripodi, Michail Kapralov, Václav Ranc, Anna Maria Timperio, Giovanna Costanzo, and Judit E. Šponer
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Circular dichroism ,Ultraviolet Rays ,Stereochemistry ,Stacking ,Context (language use) ,010402 general chemistry ,Photochemistry ,01 natural sciences ,Biochemistry ,Polymerization ,chemistry.chemical_compound ,Cyclic CMP ,Nucleotide ,Molecular Biology ,chemistry.chemical_classification ,3',5' cyclic CMP ,RNA ,nucleotides ,polymerization ,proton and UV irradiation ,Evolution, Chemical ,Oligoribonucleotides ,010405 organic chemistry ,Circular Dichroism ,Organic Chemistry ,0104 chemical sciences ,Monomer ,Models, Chemical ,chemistry ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Nucleic acid ,Molecular Medicine ,Protons - Abstract
We report that 3',5'-cyclic CMP undergoes nonenzymatic di- and trimerization at 20 °C under dry conditions upon proton or UV irradiation. The reaction involves stacking of the cyclic monomers and subsequent polymerization through serial transphosphorylations between the stacked monomers. Proton- and UV-induced oligomerization of 3',5'-cyclic CMP demonstrates that pyrimidines-similar to purines-might also have taken part in the spontaneous generation of RNA under plausible prebiotic conditions as well as in an extraterrestrial context. The observed polymerization of naturally occurring 3',5'-cyclic nucleotides supports the possibility that the extant genetic nucleic acids might have originated by way of a straight Occamian path, starting from simple reactions between plausibly preactivated monomers.
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- 2017
29. Genetically Defined Functional Modules for Spatial Orienting in the Mouse Superior Colliculus
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Nicolas Alexandre, Jérôme Boulanger, Marco Tripodi, Paula Freire-Pritchett, Laura Masullo, and Letizia Mariotti
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0301 basic medicine ,Genetic dissection ,Male ,Neurons ,Superior Colliculi ,Superior colliculus ,Modularity (biology) ,Motor control ,Sensory system ,Optogenetics ,Biology ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Glutamatergic ,Mice ,030104 developmental biology ,0302 clinical medicine ,Space Perception ,Homeobox ,Animals ,General Agricultural and Biological Sciences ,Neuroscience ,030217 neurology & neurosurgery ,Orientation, Spatial - Abstract
In order to explore and interact with their surroundings, animals need to orient toward specific positions in space. Throughout the animal kingdom, head movements represent a primary form of orienting behavior. The superior colliculus (SC) is a fundamental structure for the generation of orienting responses, but how genetically distinct groups of collicular neurons contribute to these spatially tuned behaviors remains largely to be defined. Here, through the genetic dissection of the murine SC, we identify a functionally and genetically homogeneous subclass of glutamatergic neurons defined by the expression of the paired-like homeodomain transcription factor Pitx2. We show that the optogenetic stimulation of Pitx2ON neurons drives three-dimensional head displacements characterized by stepwise, saccade-like kinematics. Furthermore, during naturalistic foraging behavior, the activity of Pitx2ON neurons precedes and predicts the onset of spatially tuned head movements. Intriguingly, we reveal that Pitx2ON neurons are clustered in an orderly array of anatomical modules that tile the entire intermediate layer of the SC. Such a modular organization gives origin to a discrete and discontinuous representation of the motor space, with each Pitx2ON module subtending a defined portion of the animal's egocentric space. The modularity of Pitx2ON neurons provides an anatomical substrate for the convergence of spatially coherent sensory and motor signals of cortical and subcortical origins, thereby promoting the recruitment of appropriate movement vectors. Overall, these data support the view of the superior colliculus as a selectively addressable and modularly organized spatial-motor register.
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- 2019
30. Éloge de la Fuite: Neural Circuits for Avoiding Dangerous Situations
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Ana González-Rueda and Marco Tripodi
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0301 basic medicine ,Behavior, Animal ,Tegmentum Mesencephali ,General Neuroscience ,Motor control ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Avoidance Learning ,Animals ,heterocyclic compounds ,Pedunculopontine Tegmental Nucleus ,Psychology ,Neuroscience ,030217 neurology & neurosurgery ,Research Articles - Abstract
An innocuous sensory stimulus that reliably signals an upcoming aversive event can be conditioned to elicit locomotion to a safe location before the aversive outcome ensues. The neural circuits that mediate the expression of this signaled locomotor action, known as signaled active avoidance, have not been identified. While exploring sensorimotor midbrain circuits in mice of either sex, we found that excitation of GABAergic cells in the substantia nigra pars reticulata blocks signaled active avoidance by inhibiting cells in the pedunculopontine tegmental nucleus (PPT), not by inhibiting cells in the superior colliculus or thalamus. Direct inhibition of putative-glutamatergic PPT cells, excitation of GABAergic PPT cells, or excitation of GABAergic afferents in PPT, abolish signaled active avoidance. Conversely, excitation of putative-glutamatergic PPT cells, or inhibition of GABAergic PPT cells, can be tuned to drive avoidance responses. The PPT is an essential junction for the expression of signaled active avoidance gated by nigral and other synaptic afferents. SIGNIFICANCE STATEMENT When a harmful situation is signaled by a sensory stimulus (e.g., street light), subjects typically learn to respond with active or passive avoidance responses that circumvent the threat. During signaled active avoidance behavior, subjects move away to avoid a threat signaled by a preceding innocuous stimulus. We identified a part of the midbrain essential to process the signal and avoid the threat. Inhibition of neurons in this area eliminates avoidance responses to the signal but preserves escape responses caused by presentation of the threat. The results highlight an essential part of the neural circuits that mediate signaled active avoidance behavior.
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- 2019
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31. YAP integrates the regulatory Snail/HNF4α circuitry controlling epithelial/hepatocyte differentiation
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Cecilia Battistelli, Laura Amicone, Raffaele Strippoli, Marco Tripodi, Veronica Consalvi, Angela Maria Cozzolino, Carla Cicchini, Alessandra Marchetti, and Valeria Noce
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Cancer Research ,Epithelial-Mesenchymal Transition ,Molecular biology ,Immunology ,Cell ,Regulator ,Down-Regulation ,Cell Cycle Proteins ,Biology ,Article ,HNF4 ,Extracellular matrix ,Mice ,Cellular and Molecular Neuroscience ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Gene silencing ,Hepatocyte ,Gene Silencing ,lcsh:QH573-671 ,Promoter Regions, Genetic ,Adaptor Proteins, Signal Transducing ,Hepatocyte differentiation ,YAP ,Snail ,EMT ,differentiation ,lcsh:Cytology ,Liver cell ,Cell Differentiation ,Epithelial Cells ,YAP-Signaling Proteins ,Cell Biology ,Up-Regulation ,Cell biology ,medicine.anatomical_structure ,Hepatocyte Nuclear Factor 4 ,Hepatocyte nuclear factor 4 ,Cell culture ,Hepatocytes ,Snail Family Transcription Factors ,Protein Binding ,Transcription Factors - Abstract
Yes-associated protein (YAP) is a transcriptional co-factor involved in many cell processes, including development, proliferation, stemness, differentiation, and tumorigenesis. It has been described as a sensor of mechanical and biochemical stimuli that enables cells to integrate environmental signals. Although in the liver the correlation between extracellular matrix elasticity (greatly increased in the most of chronic hepatic diseases), differentiation/functional state of parenchymal cells and subcellular localization/activation of YAP has been previously reported, its role as regulator of the hepatocyte differentiation remains to be clarified. The aim of this study was to evaluate the role of YAP in the regulation of epithelial/hepatocyte differentiation and to clarify how a transducer of general stimuli can integrate tissue-specific molecular mechanisms determining specific cell outcomes. By means of YAP silencing and overexpression we demonstrated that YAP has a functional role in the repression of epithelial/hepatocyte differentiation by inversely modulating the expression of Snail (master regulator of the epithelial-to-mesenchymal transition and liver stemness) and HNF4α (master regulator of hepatocyte differentiation) at transcriptional level, through the direct occupancy of their promoters. Furthermore, we found that Snail, in turn, is able to positively control YAP expression influencing protein level and subcellular localization and that HNF4α stably represses YAP transcription in differentiated hepatocytes both in cell culture and in adult liver. Overall, our data indicate YAP as a new member of the HNF4/Snail epistatic molecular circuitry previously demonstrated to control liver cell state. In this model, the dynamic balance between three main transcriptional regulators, that are able to control reciprocally their expression/activity, is responsible for the induction/maintenance of different liver cell differentiation states and its modulation could be the aim of therapeutic protocols for several chronic liver diseases.
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- 2019
32. P47 Prescripción de actividad física: 16 años después
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Luciana Grilli, Julio Szuster, Marco Tripodi, A. Sánchez, Ana Paula García, Claudio Dituro, Gustavo Ceccatto, Patricia Bernassar, Oscar Bravo, Flavia Mariel Aragón, Viviana Luján Cogo, Azul Larribité, and Lucas Sosa
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medicine.medical_specialty ,business.industry ,media_common.quotation_subject ,Cardiovascular risk factors ,Physical activity ,Odds ratio ,Confidence interval ,Feeling ,Family medicine ,medicine ,Odd ratio ,General Earth and Planetary Sciences ,Medical prescription ,business ,Statistical software ,General Environmental Science ,media_common - Abstract
Introducción: la actividad física regular contribuye al buen control metabólico, a la reducción de los factores de riesgo (FR) cardiovascular e influye en la sensación de bienestar físico, mental y social. En el año 2002, integrantes del Comité de Actividad Física de la SAD estudiaron la frecuencia de prescripción de actividad física y su asociación con factores personales en médicos asistentes al XIII Congreso Argentino de Diabetes. De los encuestados, sólo el 39,3% declaró prescribir actividad física de manera habitual. 16 años más tarde, un grupo de médicos del Capítulo Atlántico de la SAD seplanteó la necesidad de un nuevo relevamiento para determinar la situación actual.Objetivos: 1) describir la frecuencia de prescripción de actividad física por parte de médicos asistentes al XXI Congreso Argentino de Diabetes; 2) Comparar los resultados con los obtenidos en el año 2002; 3) Evaluar FR presentes en la población encuestada y su relación con la práctica de actividad físicaMateriales y métodos: se encuestaron 281 médicos (77% mujeres) asistentes al XXI Congreso Argentino de Diabetes (Mar del Plata, 2018). La encuesta evaluó la frecuencia de prescripción, la forma y factores personales del profesional que pudieran influir en su actitud: edad, sexo, FR y práctica de actividad física. Se realizaron comparaciones por estadística descriptiva entre los dos años de estudio mediante el cálculo de proporciones y representaciones gráficas. Se realizó un análisis transversal de casos y controles, calculando la razón de probabilidades. Para cada odd ratio se calculó el intervalo de confianza (IC) con P
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- 2020
33. MiR675-5p Acts on HIF-1α to Sustain Hypoxic Responses: A New Therapeutic Strategy for Glioma
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Viviana Costa, Luisa Ottobrini, Riccardo Alessandro, Cristina Martelli, Alessia Lo Dico, Cecilia Diceglie, Aroldo Rizzo, Carmine Mancone, Alice Conigliaro, Francesca Rajata, Marco Tripodi, Lo Dico, A, Costa, V, Martelli, C, Diceglie, C, Rajata, F, Rizzo, A, Mancone, C, Tripodi, M, Ottobrin,i L, Alessandro, R, and Conigliaro, A
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0301 basic medicine ,miRNA675 ,Angiogenesis ,Medicine (miscellaneous) ,RNA-binding protein ,Glioma ,HuR ,Hypoxia ,Optical imaging ,VHL ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Biology ,Toxicology and Pharmaceutics (miscellaneous) ,Cell Line ,ELAV-Like Protein 1 ,Mice ,optical imaging ,03 medical and health sciences ,Settore BIO/13 - Biologia Applicata ,Stress, Physiological ,In vivo ,microRNA ,medicine ,Animals ,Humans ,Pharmacology ,glioma ,hypoxia ,Messenger RNA ,Neovascularization, Pathologic ,RNA ,Hypoxia (medical) ,Hypoxia-Inducible Factor 1, alpha Subunit ,medicine.disease ,3. Good health ,Angiogenesi ,MicroRNAs ,030104 developmental biology ,Immunology ,Cancer research ,Heterografts ,medicine.symptom ,Research Paper - Abstract
Hypoxia is a common feature in solid tumours. In glioma, it is considered the major driving force for tumour angiogenesis and correlates with enhanced resistance to conventional therapies, increased invasiveness and a poor prognosis for patients. Here we describe, for the first time, that miR675-5p, embedded in hypoxia-induced long non-coding RNA H19, plays a mandatory role in establishing a hypoxic response and in promoting hypoxia-mediated angiogenesis. We demonstrated, in vitro and in vivo, that miR675-5p over expression in normoxia is sufficient to induce a hypoxic moreover, miR675-5p depletion in low oxygen conditions, drastically abolishes hypoxic responses including angiogenesis. In addition, our data indicate an interaction of miR675-5p, HIF-1α mRNA and the RNA Binding Protein HuR in hypoxia-induced responses. We suggest the modulation of miR675-5p as a new therapeutic option to promote or abolish hypoxia induced angiogenesis.
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- 2016
34. Development of alkyl glycerone phosphate synthase inhibitors: Structure-activity relationship and effects on ether lipids and epithelial-mesenchymal transition in cancer cells
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Giulia Stazi, Sharon M. Louie, Sara Marchese, Giancarlo Fabrizi, Andrea Mattevi, Roberto Cirilli, Valentina Piano, Monica Viviano, Rino Ragno, Alexandros Patsilinakos, Cecilia Battistelli, Alessia Ciogli, Raffaele Strippoli, Antonello Mai, Lorenzo Antonini, Marco Tripodi, Alessandra Marchetti, Roberta Mazzone, Daniel K. Nomura, Biagina Marrocco, Gianluca Sbardella, Clemens Zwergel, and Sergio Valente
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Epithelial-Mesenchymal Transition ,Motility ,01 natural sciences ,Cell Line ,03 medical and health sciences ,Structure-Activity Relationship ,Cell Movement ,Cell Line, Tumor ,Neoplasms ,Drug Discovery ,medicine ,AGPS inhibitors ,Cancer ,E-cadherin ,Ether lipids ,Snail ,Humans ,Epithelial–mesenchymal transition ,030304 developmental biology ,Cell Proliferation ,Pharmacology ,0303 health sciences ,Alkyl and Aryl Transferases ,010405 organic chemistry ,Cell growth ,Chemistry ,Organic Chemistry ,Lipid metabolism ,Cell migration ,General Medicine ,medicine.disease ,Cadherins ,Lipid Metabolism ,0104 chemical sciences ,Cell biology ,Cell culture ,Cancer cell ,Matrix Metalloproteinase 2 ,Snail Family Transcription Factors - Abstract
In aggressive tumors, alkylglyceronephosphate synthase (AGPS) controls cellular ether phospholipid utilization and metabolism to promote cancer cell proliferation and motility. SAR studies on the first-in-class AGPS inhibitor 1, discovered by our group, led to the 2,6-difluoro analog 2i which showed higher binding affinity than 1 in vitro. In 231MFP cancer cells, 2i reduced ether lipids levels and cell migration rate. When tested in PC-3 and MDA-MB-231 cancer cells, 2i specifically impaired epithelial to mesenchymal transition (EMT) by modulating E-cadherin, Snail and MMP2 expression levels. Moreover, the combination of siRNAs against AGPS and 2i provided no additive effect, confirming that the modulation of 2i on EMT specifically relies on AGPS inhibition. Finally, this compound also affected cancer cell proliferation especially in MDA-MB-231 cells expressing higher AGPS level, whereas it provided negligible effects on MeT5A, a non-tumorigenic cell line, thus showing cancer specificity.
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- 2018
35. Cerebral organoids at the air-liquid interface generate diverse nerve tracts with functional output
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Laura Masullo, Madeline A. Lancaster, Marco Tripodi, Timothy Sit, Stefano L. Giandomenico, Magdalena Sutcliffe, Jérôme Boulanger, Léa M. D. Wenger, Susanna B. Mierau, Ole Paulsen, Emmanuel Derivery, George M. Gibbons, Andras Lakatos, Mierau, Susanna [0000-0002-1220-6423], Paulsen, Ole [0000-0002-2258-5455], Lakatos, Andras [0000-0002-1301-2292], and Apollo - University of Cambridge Repository
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Male ,Pluripotent Stem Cells ,0301 basic medicine ,Decussation ,corticospinal tract ,brain and spinal cord injury ,Cell Survival ,organoid ,Cellular differentiation ,medicine.medical_treatment ,Biology ,Article ,corpus callosum ,Tissue Culture Techniques ,03 medical and health sciences ,0302 clinical medicine ,Neural Pathways ,medicine ,Organoid ,Humans ,nerve tracts ,Axon ,Growth cone ,030304 developmental biology ,Cerebral Cortex ,Neurons ,0303 health sciences ,Axon guidance ,General Neuroscience ,Neurogenesis ,in vitro ,Human brain ,Retrograde tracing ,Axons ,Organoids ,030104 developmental biology ,medicine.anatomical_structure ,Cerebral cortex ,cerebral organoids ,Nerve tract ,Female ,Axotomy ,medicine.symptom ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Three-dimensional neural organoids are emerging tools with the potential for improving our understanding of human brain development and neurological disorders. Recent advances in this field have demonstrated their capacity to model neurogenesis1,2, neuronal migration and positioning3,4, and even response to sensory input5. However, it remains to be seen whether these tissues can model axon guidance dynamics and the formation of complex connectivity with functional neuronal output. Here, we have established a longterm air-liquid interface culture paradigm that leads to improved neuronal survival and allows for imaging of axon guidance. Over time, these cultures spontaneously form thick axon tracts capable of projecting over long distances. Axon bundles display various morphological behaviors including intracortical projection within and across the organoid, growth cone turning, decussation, and projection away from the organoid. Single-cell RNA-sequencing reveals the full repertoire of cortical neuronal identities, and retrograde labelling demonstrates these tract morphologies match the appropriate molecular identities, namely callosal and corticofugal neuron types. We show that these neurons are functionally mature, generate active networks within the organoid, and that extracortical projecting tracts innervate and activate mouse spinal cord-muscle explants. Muscle contractions can be evoked by stimulation of the organoid, while axotomy of the innervating tracts abolishes the muscle contraction response, demonstrating dependence on connection with the organoid. Overall, these results reveal a remarkable selforganization of corticofugal and callosal tracts with a functional output, providing new opportunities to examine relevant aspects of human CNS development and response to injury.
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- 2018
36. A cryptic RNA-binding domain mediates Syncrip recognition and exosomal partitioning of miRNA targets
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Evangelos Christodoulou, Andres Ramos, Roksana W. Ogrodowicz, André Dallmann, Cecilia Battistelli, Fruzsina Hobor, Ian A. Taylor, Stephen R. Martin, Neil J. Ball, Marco Tripodi, Alfredo Castello, and Carla Cicchini
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0301 basic medicine ,Models, Molecular ,Protein Conformation, alpha-Helical ,Science ,Genetic Vectors ,General Physics and Astronomy ,Gene Expression ,Sequence alignment ,MiRNA binding ,Computational biology ,Plasma protein binding ,Biology ,Crystallography, X-Ray ,Exosomes ,General Biochemistry, Genetics and Molecular Biology ,miRnas. RNa binding protein ,Article ,Heterogeneous-Nuclear Ribonucleoproteins ,03 medical and health sciences ,Protein structure ,microRNA ,Escherichia coli ,Animals ,Drosophila Proteins ,Humans ,Protein Interaction Domains and Motifs ,Amino Acid Sequence ,Cloning, Molecular ,lcsh:Science ,Peptide sequence ,Multidisciplinary ,Binding Sites ,Sequence Homology, Amino Acid ,RNA ,RNA-Binding Proteins ,General Chemistry ,Aptamers, Nucleotide ,Recombinant Proteins ,MicroRNAs ,030104 developmental biology ,Drosophila melanogaster ,lcsh:Q ,Protein Conformation, beta-Strand ,Sequence Alignment ,Binding domain ,Protein Binding - Abstract
Exosomal miRNA transfer is a mechanism for cell–cell communication that is important in the immune response, in the functioning of the nervous system and in cancer. Syncrip/hnRNPQ is a highly conserved RNA-binding protein that mediates the exosomal partition of a set of miRNAs. Here, we report that Syncrip’s amino-terminal domain, which was previously thought to mediate protein–protein interactions, is a cryptic, conserved and sequence-specific RNA-binding domain, designated NURR (N-terminal unit for RNA recognition). The NURR domain mediates the specific recognition of a short hEXO sequence defining Syncrip exosomal miRNA targets, and is coupled by a non-canonical structural element to Syncrip’s RRM domains to achieve high-affinity miRNA binding. As a consequence, Syncrip-mediated selection of the target miRNAs implies both recognition of the hEXO sequence by the NURR domain and binding of the RRM domains 5′ to this sequence. This structural arrangement enables Syncrip-mediated selection of miRNAs with different seed sequences., Syncrip is a conserved RNA-binding protein that mediates selective miRNA loading into cell-secreted exosomes, which is important for cell–cell communication. Here the authors provide mechanistic insights into how Syncrip recognises its target miRNAs by combining NMR and crystallography.
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- 2018
37. Three-Dimensional Representation of Motor Space in the Mouse Superior Colliculus
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Marco Tripodi, Jonathan J. Wilson, Caterina Trentin, and Nicolas Alexandre
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0301 basic medicine ,Male ,Superior Colliculi ,Rotation ,Biology ,Space (mathematics) ,General Biochemistry, Genetics and Molecular Biology ,Displacement (vector) ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Animals ,Neurons ,Landmark ,Superior colliculus ,Representation (systemics) ,Motor control ,Electrophysiological Phenomena ,Mice, Inbred C57BL ,Electrophysiology ,030104 developmental biology ,Space Perception ,Metric (mathematics) ,General Agricultural and Biological Sciences ,Neuroscience ,Head ,030217 neurology & neurosurgery - Abstract
From the act of exploring an environment to that of grasping a cup of tea, animals must put in register their motor acts with their surrounding space. In the motor domain, this is likely to be defined by a register of three-dimensional (3D) displacement vectors, whose recruitment allows motion in the direction of a target. One such spatially targeted action is seen in the head reorientation behavior of mice, yet the neural mechanisms underlying these 3D behaviors remain unknown. Here, by developing a head-mounted inertial sensor for studying 3D head rotations and combining it with electrophysiological recordings, we show that neurons in the mouse superior colliculus are either individually or conjunctively tuned to the three Eulerian components of head rotation. The average displacement vectors associated with motor-tuned colliculus neurons remain stable over time and are unaffected by changes in firing rate or the duration of spike trains. Finally, we show that the motor tuning of collicular neurons is largely independent from visual or landmark cues. By describing the 3D nature of motor tuning in the superior colliculus, we contribute to long-standing debate on the dimensionality of collicular motor decoding; furthermore, by providing an experimental paradigm for the study of the metric of motor tuning in mice, this study also paves the way to the genetic dissection of the circuits underlying spatially targeted motion.
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- 2018
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38. SMO inhibition modulates cellular plasticity and invasiveness in colorectal cancer
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Paolo Magistri, Marco Tripodi, Lucia Rossi, Niccolò Petrucciani, Francesco D'Angelo, Teijo Pellinen, Giovanni Ramacciato, Paolo Aurello, Livia Maria Mangogna, Giuseppe Nigri, Cecilia Battistelli, Raffaele Strippoli, Institute for Molecular Medicine Finland, and Precision Systems Medicine
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0301 basic medicine ,CARCINOMA ,Biology ,cell invasiveness ,MECHANISMS ,ACTIVATION ,03 medical and health sciences ,0302 clinical medicine ,hedgehog pathway ,Downregulation and upregulation ,GLI1 ,GLI2 ,Pharmacology (medical) ,Epithelial–mesenchymal transition ,Autocrine signalling ,Hedgehog ,Original Research ,Pharmacology ,SMO inhibition ,cell plasticity ,colon cancer ,EMT ,pharmacology ,TUMOR-GROWTH ,COLON-CANCER ,lcsh:RM1-950 ,PROLIFERATION ,IN-VITRO ,HEDGEHOG SIGNALING PATHWAY ,EPITHELIAL-MESENCHYMAL TRANSITION ,Hedgehog signaling pathway ,3. Good health ,030104 developmental biology ,lcsh:Therapeutics. Pharmacology ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,3111 Biomedicine ,Smoothened ,STEM-CELLS - Abstract
HIGHLIGHTS Preliminary results of this work were presented at the 2016 Academic Surgical Congress, Jacksonville (FL), February 2-4 2016 (Original title: Selective Smo-Inhibition Interferes With Cellular Energetic Metabolism In Colorectal Cancer)This study was funded by "Sapienza-University of Rome" (Funds for young researchers) and "AIRC" (Italian Association for Cancer Research)Hedgehog inhibitor was kindly provided by Genentech, Inc.®. Colon Cancer (CC) is the fourth most frequently diagnosed tumor and the second leading cause of death in the USA. Abnormalities of Hedgehog pathway have been demonstrated in several types of human cancers, however the role of Hedgehog (Hh) in CC remain controversial. In this study, we analyzed the association between increased mRNA expression of GLI1 and GLI2, two Hh target genes, and CC survival and recurrence by gene expression microarray from a cohort of 382 CC patients. We found that patients with increased expression of GLI1 showed a statistically significant reduction in survival. In order to demonstrate a causal role of Hh pathway activation in the pathogenesis of CC, we treated HCT 116, SW480 and SW620 CC cells lines with GDC-0449, a pharmacological inhibitor of Smoothened (SMO). Treatment with GDC-0449 markedly reduced expression of Hh target genes GLI1, PTCH1, HIP1, MUC5AC, thus indicating that this pathway is constitutively active in CC cell lines. Moreover, GDC-0449 partially reduced cell proliferation, which was associated with upregulation of p21 and downregulation of CycD1. Finally, treatment with the same drug reduced migration and three-dimensional invasion, which were associated with downregulation of Snail1, the EMT master gene, and with induction of the epithelial markers Cytokeratin-18 and E-cadherin. These results were confirmed by SMO genetic silencing. Notably, treatment with 5E1, a Sonic Hedgehog-specific mAb, markedly reduced the expression of Hedgehog target genes, as well as inhibited cell proliferation and mediated reversion toward an epithelial phenotype. This suggests the existence of a Hedgehog autocrine signaling loop affecting cell plasticity and fostering cell proliferation and migration/invasion in CC cell lines. These discoveries encourage future investigations to better characterize the role of Hedgehog in cellular plasticity and invasion during the different steps of CC pathogenesis.
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- 2018
39. HDAC1 inhibition by MS-275 in mesothelial cells limits cellular invasion and promotes MMT reversal
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Antonello Mai, Lucia Rossi, Alessandro Domenici, Cecilia Battistelli, Raffaele Strippoli, Clemens Zwergel, Valeria Noce, Veronica Bordoni, Sergio Valente, Marco Tripodi, Alessandra Moioli, Paolo Menè, Valeria de Turris, Andrea Manzione, and Marco Palladino
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0301 basic medicine ,Male ,MMP2 ,Epithelial-Mesenchymal Transition ,Pyridines ,Science ,Histone Deacetylase 1 ,Occludin ,Article ,Transforming Growth Factor beta1 ,03 medical and health sciences ,Downregulation and upregulation ,HDAC inhibitors ,Cell Movement ,EMT, fibrosis, gene expression regulation ,Gene silencing ,Humans ,RNA, Small Interfering ,Histone H3 acetylation ,Peritoneal Fibrosis ,Aged ,Aged, 80 and over ,Multidisciplinary ,Chemistry ,fibrosis ,EMT ,Epithelial Cells ,gene expression regulation ,Middle Aged ,Cadherins ,HDAC1 ,Histone Deacetylase Inhibitors ,030104 developmental biology ,Benzamides ,Cancer research ,Kidney Failure, Chronic ,Medicine ,Female ,RNA Interference ,Histone deacetylase ,Snail Family Transcription Factors ,Peritoneum ,Peritoneal Dialysis - Abstract
Peritoneal fibrosis is a pathological alteration of the peritoneal membrane occurring in a variety of conditions including peritoneal dialysis (PD), post-surgery adhesions and peritoneal metastases. The acquisition of invasive and pro-fibrotic abilities by mesothelial cells (MCs) through induction of MMT, a cell-specific form of EMT, plays a main role in this process. Aim of this study was to evaluate possible effects of histone deacetylase (HDAC) inhibitors, key components of the epigenetic machinery, in counteracting MMT observed in MCs isolated from effluent of PD patients. HDAC inhibitors with different class/isoform selectivity have been used for pharmacological inhibition. While the effect of other inhibitors was limited to a partial E-cadherin re-expression, MS-275, a HDAC1-3 inhibitor, promoted: (i) downregulation of mesenchymal markers (MMP2, Col1A1, PAI-1, TGFβ1, TGFβRI) (ii) upregulation of epithelial markers (E-cadherin, Occludin), (iii) reacquisition of an epithelial-like morphology and (iv) marked reduction of cellular invasiveness. Results were confirmed by HDAC1 genetic silencing. Mechanistically, MS-275 causes: (i) increase of nuclear histone H3 acetylation (ii) rescue of the acetylation profile on E-cadherin promoter, (iii) Snail functional impairment. Overall, our study, pinpointing a role for HDAC1, revealed a new player in the regulation of peritoneal fibrosis, providing the rationale for future therapeutic opportunities.
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- 2018
40. Hepatitis C virus direct-acting antivirals therapy impacts on extracellular vesicles microRNAs content and on their immunomodulating properties
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Claudia Montaldo, Marco Tripodi, Alessandra Zingoni, Cecilia Battistelli, Veronica Bordoni, Gianpiero D'Offizi, Laura Santangelo, Angela Santoni, Eleonora Cimini, Chiara Agrati, and Maria Rosaria Capobianchi
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Adult ,Male ,0301 basic medicine ,Hepatitis C virus ,Cell Communication ,Hepacivirus ,Exosomes ,medicine.disease_cause ,Antiviral Agents ,Exosome ,Natural Killer cell ,Natural killer cell ,Pathogenesis ,03 medical and health sciences ,Immune system ,medicine ,Humans ,Direct-acting antiviral agents ,MicroRNA ,Hepatology ,Aged ,Innate immune system ,business.industry ,Gene Expression Profiling ,Degranulation ,Hepatitis C, Chronic ,Middle Aged ,Immunity, Innate ,Microvesicles ,MicroRNAs ,030104 developmental biology ,medicine.anatomical_structure ,Case-Control Studies ,Immunology ,Female ,business ,Biomarkers - Abstract
Background & aims Hepatitis C virus (HCV) infection is known to cause major alterations in the cross-talk between hepatic and immune cells thus contributing to the liver disease pathogenesis. Extracellular vesicles have been proved to act as major players in cell-cell communication, and their cargo changes in relation to pathophysiological states. The aim of this study was to evaluate the effects of chronic HCV infection and direct-acting antivirals (DAA) on exosome-delivered microRNAs and on their ability to modulate the innate immune response. Methods Exosomes isolated from the plasma of healthy donors and naive, viremic HCV patients before and after DAA treatment have been compared for their microRNAs cargo by quantitative polymerase chain reaction. Functional assays with peripheral blood cells from healthy donors were performed to assess exosome-mediated immune responses. Results MicroRNAs associated with HCV-related immunopathogenesis which were found to be enriched in exosomes of HCV viremic patients (in particular, miR-122-5p, miR-222-3p, miR-146a, miR-150-5p, miR-30c, miR-378a-3p and miR-20a-5p) were markedly reduced by DAA therapy. This exosome-microRNA cargo modulation parallels changes in their immunomodulatory properties in ex vivo experiments. Exosomes from HCV patients inhibit NK degranulation activity and this effect correlates with miR-122-5p or miR-222-3p levels. Conclusions Enrichment of immunomodulatory microRNAs in exosomes of HCV patients was correlated with their inhibitory activity on innate immune cells function. Direct-acting antivirals (DAA) treatment was observed to revert both microRNA content and functional profiles of systemic exosomes towards those of healthy donors. Exosome-associated microRNAs may provide valuable biomarkers to monitor immune response recovery.
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- 2018
41. Labeling and identifying cell-type-specific proteomes in the mouse brain
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Toke P. Krogager, Russell J. Ernst, Michael H. Hastings, Václav Beránek, Marco Tripodi, Jason W. Chin, Laura Calo, Maria Grazia Spillantini, Ernesto Ciabatti, Thomas S. Elliott, Spillantini, Maria [0000-0002-8544-7332], Hastings, Michael [0000-0001-8576-6651], Chin, Jason [0000-0003-1219-4757], and Apollo - University of Cambridge Repository
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0301 basic medicine ,Cell type ,Proteome ,Biomedical Engineering ,Bioengineering ,Biology ,Applied Microbiology and Biotechnology ,Article ,Gene Expression Regulation, Enzymologic ,Amino Acyl-tRNA Synthetases ,03 medical and health sciences ,Mice ,0302 clinical medicine ,RNA, Transfer ,Cortex (anatomy) ,medicine ,Animals ,Amino Acids ,Regulation of gene expression ,Cell specific ,chemistry.chemical_classification ,Neurons ,RNA ,Brain ,3. Good health ,Amino acid ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Molecular Medicine ,Neuroglia ,030217 neurology & neurosurgery ,Biotechnology - Abstract
We develop an approach to tag proteomes synthesized by specific cell types in dissociated cortex, brain slices, and the brains of live mice. By viral-mediated expression of an orthogonal pyrrolysyl-tRNA synthetase-tRNAXXX pair in a cell type of interest and providing a non-canonical amino acid with a chemical handle, we selectively label neuronal or glial proteomes. The method enables the identification of proteins from spatially and genetically defined regions of the brain.
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- 2017
42. Negative Regulation of Mitochondrial Antiviral Signaling Protein-Mediated Antiviral Signaling by the Mitochondrial Protein LRPPRC During Hepatitis C Virus Infection
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Mauro Piacentini, Fabiola Ciccosanti, Marco Tripodi, Giuseppe Ippolito, Giulia Refolo, Martina Di Rienzo, Ariel Basulto Perdomo, Tonino Alonzi, Gian Maria Fimia, and Marta Romani
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0301 basic medicine ,TRAF3 ,viruses ,Hepacivirus ,Biology ,Viral Nonstructural Proteins ,Mitochondrial Proteins ,03 medical and health sciences ,0302 clinical medicine ,Downregulation and upregulation ,Interferon ,medicine ,Humans ,NS5A ,Cells, Cultured ,Mitochondrial antiviral-signaling protein ,Adaptor Proteins, Signal Transducing ,Innate immune system ,Hepatology ,virus diseases ,biochemical phenomena, metabolism, and nutrition ,Hepatitis C, Chronic ,Virology ,digestive system diseases ,Neoplasm Proteins ,030104 developmental biology ,Viral replication ,030211 gastroenterology & hepatology ,Signal transduction ,medicine.drug ,Signal Transduction - Abstract
Hepatitis C virus (HCV) is highly efficient in establishing a chronic infection, having evolved multiple strategies to suppress the host antiviral responses. The HCV nonstructural 5A (NS5A) protein, in addition to its role in viral replication and assembly, has long been known to hamper the interferon (IFN) response. However, the mechanism of this inhibitory activity of NS5A remains partly characterized. In a functional proteomic screening carried out in HCV replicon cells, we identified the mitochondrial protein LRPPRC as an NS5A binding factor. Notably, we found that downregulation of LRPPRC expression results in a significant inhibition of HCV infection, which is associated with an increased activation of the IFN response. Moreover, we showed that LRPPRC acts as a negative regulator of the mitochondrial-mediated antiviral immunity, by interacting with mitochondrial antiviral signaling protein (MAVS) and inhibiting its association with TRAF3 and TRAF6. Finally, we demonstrated that NS5A is able to interfere with MAVS activity in a LRPPRC-dependent manner. Conclusion: Overall, our results indicate that NS5A contributes to the inhibition of innate immune pathways during HCV infection by exploiting the ability of LRPPRC to inhibit MAVS-regulated antiviral signaling.
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- 2017
43. New Tools for Molecular Therapy of Hepatocellular Carcinoma
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Angela Maria Cozzolino, Marco Tripodi, Alessandra Marchetti, and Francesca Bisceglia
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Hepatocyte differentiation ,Cirrhosis ,LETFs ,Genetic enhancement ,lcsh:R ,EMT ,lcsh:Medicine ,Review ,Biology ,HCC ,HNF4α ,miRNAs ,TGFβ ,gene therapy ,medicine.disease ,digestive system diseases ,Targeted Molecular Therapy ,Hepatocellular carcinoma ,Immunology ,medicine ,Cancer research ,Neoplastic transformation ,Liver cancer ,Reprogramming - Abstract
Hepatocellular carcinoma (HCC) is the most common type of liver cancer, arising from neoplastic transformation of hepatocytes or liver precursor/stem cells. HCC is often associated with pre-existing chronic liver pathologies of different origin (mainly subsequent to HBV and HCV infections), such as fibrosis or cirrhosis. Current therapies are essentially still ineffective, due both to the tumor heterogeneity and the frequent late diagnosis, making necessary the creation of new therapeutic strategies to inhibit tumor onset and progression and improve the survival of patients. A promising strategy for treatment of HCC is the targeted molecular therapy based on the restoration of tumor suppressor proteins lost during neoplastic transformation. In particular, the delivery of master genes of epithelial/hepatocyte differentiation, able to trigger an extensive reprogramming of gene expression, could allow the induction of an efficient antitumor response through the simultaneous adjustment of multiple genetic/epigenetic alterations contributing to tumor development. Here, we report recent literature data supporting the use of members of the liver enriched transcription factor (LETF) family, in particular HNF4α, as tools for gene therapy of HCC.
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- 2015
44. Epigenetic control of EMT/MET dynamics: HNF4α impacts DNMT3s through miRs-29
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Laura Amicone, Marco De Santis Puzzonia, Angela Maria Cozzolino, Valeria de Nonno, Marco Tripodi, Silvia Anna Ciafrè, Chad Brocker, Cecilia Battistelli, Carla Cicchini, and Frank J. Gonzalez
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miR-29 ,Epithelial-Mesenchymal Transition ,Epigenetic code ,DNMT3B ,Biophysics ,De novo DNA methylation ,Biology ,Biochemistry ,DNA methyltransferase ,Article ,Gene Expression Regulation, Enzymologic ,DNA Methyltransferase 3A ,Epigenesis, Genetic ,TGFβ ,Mice ,Structural Biology ,microRNA ,Genetics ,Animals ,Hepatocyte ,DNA (Cytosine-5-)-Methyltransferases ,Epigenetics ,Molecular Biology ,Epithelial–mesenchymal transition ,Cells, Cultured ,Mice, Knockout ,Settore BIO/13 ,Epithelial-mesenchymal transition ,MiR-29 ,Cell Differentiation ,Cellular Reprogramming ,Cell biology ,Gene Expression Regulation, Neoplastic ,MicroRNAs ,Cell Transformation, Neoplastic ,Hepatocyte Nuclear Factor 4 ,embryonic structures ,DNA methylation ,Hepatocytes ,DNMT1 ,Reprogramming - Abstract
Background and aims Epithelial-to-mesenchymal transition (EMT) and the reverse mesenchymal-to-epithelial transition (MET) are manifestations of cellular plasticity that imply a dynamic and profound gene expression reprogramming. While a major epigenetic code controlling the coordinated regulation of a whole transcriptional profile is guaranteed by DNA methylation, DNA methyltransferase (DNMT) activities in EMT/MET dynamics are still largely unexplored. Here, we investigated the molecular mechanisms directly linking HNF4α, the master effector of MET, to the regulation of both de novo of DNMT 3A and 3B. Methods Correlation among EMT/MET markers, microRNA29 and DNMT3s expression was evaluated by RT-qPCR, Western blotting and immunocytochemical analysis. Functional roles of microRNAs and DNMT3s were tested by anti-miRs, microRNA precursors and chemical inhibitors. ChIP was utilized for investigating HNF4α DNA binding activity. Results HNF4α silencing was sufficient to induce positive modulation of DNMT3B, in in vitro differentiated hepatocytes as well as in vivo hepatocyte-specific Hnf4α knockout mice, and DNMT3A, in vitro, but not DNMT1. In exploring the molecular mechanisms underlying these observations, evidence have been gathered for (i) the inverse correlation between DNMT3 levels and the expression of their regulators miR-29a and miR-29b and (ii) the role of HNF4α as a direct regulator of miR-29a-b transcription. Notably, during TGFβ-induced EMT, DNMT3s' pivotal function has been proved, thus suggesting the need for the repression of these DNMTs in the maintenance of a differentiated phenotype. Conclusions HNF4α maintains hepatocyte identity by regulating miR-29a and -29b expression, which in turn control epigenetic modifications by limiting DNMT3A and DNMT3B levels.
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- 2015
45. Iron overload down-regulates the expression of the HIV-1 Rev cofactor eIF5A in infected T lymphocytes
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Dario Benelli, Marco Tripodi, Giulia Refolo, Alessio Grimaldi, Mauro Piacentini, Vincenzo Barnaba, Gabriella Rozera, Gian Maria Fimia, Carmine Mancone, Fabiola Ciccosanti, and Isabella Abbate
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0301 basic medicine ,Settore BIO/06 ,lcsh:Cytology ,Research ,030106 microbiology ,Quantitative proteomics ,HIV-1 infection ,Iron overload ,Spike-in SILAC ,Biology ,Proteomics ,Biochemistry ,Virology ,Small hairpin RNA ,03 medical and health sciences ,030104 developmental biology ,Downregulation and upregulation ,Stable isotope labeling by amino acids in cell culture ,Gene expression ,Proteome ,lcsh:QH573-671 ,Molecular Biology ,EIF5A - Abstract
Background Changes in iron metabolism frequently accompany HIV-1 infection. However, while many clinical and in vitro studies report iron overload exacerbates the development of infection, many others have found no correlation. Therefore, the multi-faceted role of iron in HIV-1 infection remains enigmatic. Methods RT-qPCR targeting the LTR region, gag, Tat and Rev were performed to measure the levels of viral RNAs in response to iron overload. Spike-in SILAC proteomics comparing i) iron-treated, ii) HIV-1-infected and iii) HIV-1-infected/iron treated T lymphocytes was performed to define modifications in the host cell proteome. Data from quantitative proteomics were integrated with the HIV-1 Human Interaction Database for assessing any viral cofactors modulated by iron overload in infected T lymphocytes. Results Here, we demonstrate that the iron overload down-regulates HIV-1 gene expression by decreasing the levels of viral RNAs. In addition, we found that iron overload modulates the expression of many viral cofactors. Among them, the downregulation of the REV cofactor eIF5A may correlate with the iron-induced inhibition of HIV-1 gene expression. Therefore, we demonstrated that eiF5A downregulation by shRNA resulted in a significant decrease of Nef levels, thus hampering HIV-1 replication. Conclusions Our study indicates that HIV-1 cofactors influenced by iron metabolism represent potential targets for antiretroviral therapy and suggests eIF5A as a selective target for drug development. Electronic supplementary material The online version of this article (doi:10.1186/s12953-017-0126-0) contains supplementary material, which is available to authorized users.
- Published
- 2017
46. Life-Long Genetic and Functional Access to Neural Circuits Using Self-Inactivating Rabies Virus
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Marco Tripodi, Ernesto Ciabatti, Letizia Mariotti, Ana González-Rueda, and Fabio Morgese
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0301 basic medicine ,Resource ,rabies ,Biology ,Optogenetics ,optogenetic ,medicine.disease_cause ,General Biochemistry, Genetics and Molecular Biology ,Virus ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Neurobiology ,Neural Pathways ,Biological neural network ,medicine ,Animals ,Functional studies ,neural circuits ,Neurons ,Artificial neural network ,Rabies virus ,in vivo imaging ,DNA ,Network dynamics ,medicine.disease ,network activity ,3. Good health ,transsynaptic tracing ,circuit manipulations ,030104 developmental biology ,Immunology ,Synapses ,Rabies ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Summary Neural networks are emerging as the fundamental computational unit of the brain and it is becoming progressively clearer that network dysfunction is at the core of a number of psychiatric and neurodegenerative disorders. Yet, our ability to target specific networks for functional or genetic manipulations remains limited. Monosynaptically restricted rabies virus facilitates the anatomical investigation of neural circuits. However, the inherent cytotoxicity of the rabies largely prevents its implementation in long-term functional studies and the genetic manipulation of neural networks. To overcome this limitation, we developed a self-inactivating ΔG-rabies virus (SiR) that transcriptionally disappears from the infected neurons while leaving permanent genetic access to the traced network. SiR provides a virtually unlimited temporal window for the study of network dynamics and for the genetic and functional manipulation of neural circuits in vivo without adverse effects on neuronal physiology and circuit function., Graphical Abstract, Highlights • Self-inactivating rabies virus (SiR) provides life-long access to neural networks • SiR has no adverse effects on neuronal physiology and network computations • SiR allows open-ended Ca2+ imaging of network activity and functional intervention • SiR allows generation of circuit-specific knockouts, A self-inactivating rabies virus provides a virtually unlimited temporal window for the study and manipulation of neural circuits in vivo without adverse effects on neuronal physiology and circuit function.
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- 2017
47. The laminA/NF-Y protein complex reveals an unknown transcriptional mechanism on cell proliferation
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Giulia Regazzo, Paolo Ciana, Aymone Gurtner, Maurizio C. Capogrossi, Lucia Cicchillitti, Carmine Mancone, Manuela Spagnuolo, Marco Tripodi, Fabrizio Carlomosti, Giulia Dell'Omo, Maria Lucia Dell'Anna, Tonino Alonzi, Giulia Piaggio, Isabella Manni, Mauro Picardo, Alessandra Magenta, and Maria Giulia Rizzo
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0301 basic medicine ,Chromatin Immunoprecipitation ,Euchromatin ,Transcription, Genetic ,Proliferation ,Mice, Transgenic ,Biology ,Cell cycle ,Response Elements ,03 medical and health sciences ,Mice ,Cell Line, Tumor ,Animals ,Humans ,Promoter Regions, Genetic ,Cell Proliferation ,Nuclear lamina ,Transcription ,Oncology ,Genetics ,Regulation of gene expression ,Transcriptional activity ,CCAAT-Binding-Factor ,Cell growth ,High-Throughput Nucleotide Sequencing ,Lamin Type A ,humanities ,DNA-Binding Proteins ,Protein Transport ,030104 developmental biology ,CCAAT-Binding Factor ,Gene Expression Regulation ,Multiprotein Complexes ,Lamin ,Research Paper ,Protein Binding - Abstract
// Lucia Cicchillitti 1, * , Isabella Manni 1, * , Carmine Mancone 2, 3 , Giulia Regazzo 4 , Manuela Spagnuolo 4 , Tonino Alonzi 2 , Fabrizio Carlomosti 5, † , Maria Lucia Dell’Anna 6 , Giulia Dell’Omo 7 , Mauro Picardo 6 , Paolo Ciana 8 , Maurizio C. Capogrossi 5 , Marco Tripodi 2, 3 , Alessandra Magenta 5 , Maria Giulia Rizzo 4 , Aymone Gurtner 1, # , Giulia Piaggio 1, # 1 Department of Research, Advanced Diagnostics and Technological Innovation, SAFU Unit, Translational Research Area, Regina Elena National Cancer Institute, 00144 Rome, Italy 2 National Institute for Infectious Diseases L. Spallanzani, IRCCS, Department of Epidemiology and Preclinical Research, 00149 Rome, Italy 3 Department of Cellular Biotechnologies and Haematology, Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Sapienza University of Rome, 00161 Rome, Italy 4 Department of Research, Advanced Diagnostics and Technological Innovation, Genomic and Epigenetic Unit, Translational Research Area, Regina Elena National Cancer Institute, Rome, Italy 5 Fondazione Luigi Maria Monti, Istituto Dermopatico dell’Immacolata-IRCCS, Laboratorio di Patologia Vascolare, 00167 Rome, Italy 6 Cutaneous Physiopathology and Integrated Center of Metabolomics Research, San Gallicano Dermatologic Institute, IRCCS, 00144 Rome, Italy 7 Department of Oncology and Hemato-Oncology and Department of Pharmacological and Biomolecular Sciences, University of Milan, 20133 Milan, Italy 8 Center of Excellence on Neurodegenerative Diseases, Department of Oncology and Hemato-Oncology, University of Milan, 20133 Milan, Italy * These authors have contributed equally to this work # Co-last authors † In memory of Fabrizio Carlomosti who tragically passed away on October the 29 th 2015 Correspondence to: Giulia Piaggio, email: piaggio@ifo.it Aymone Gurtner, email: gurtner@ifo.it Keywords: transcription, proliferation, cell cycle, euchromatin, nuclear lamina Received: May 18, 2016 Accepted: October 10, 2016 Published: October 26, 2016 ABSTRACT Lamin A is a component of the nuclear matrix that also controls proliferation by largely unknown mechanisms. NF-Y is a ubiquitous protein involved in cell proliferation composed of three subunits (-YA -YB -YC) all required for the DNA binding and transactivation activity. To get clues on new NF-Y partner(s) we performed a mass spectrometry screening of proteins that co-precipitate with the regulatory subunit of the complex, NF-YA. By this screening we identified lamin A as a novel putative NF-Y interactor. Co-immunoprecipitation experiments and confocal analysis confirmed the interaction between the two endogenous proteins. Interestingly, this association occurs on euchromatin regions, too. ChIP experiments demonstrate lamin A enrichment in several promoter regions of cell cycle related genes in a NF-Y dependent manner. Gain and loss of function experiments reveal that lamin A counteracts NF-Y transcriptional activity. Taking advantage of a recently generated transgenic reporter mouse, called MITO-Luc, in which an NF-Y–dependent promoter controls luciferase expression, we demonstrate that lamin A counteracts NF-Y transcriptional activity not only in culture cells but also in living animals. Altogether, our data demonstrate the occurrence of lamin A/NF-Y interaction and suggest a possible role of this protein complex in regulation of NF-Y function in cell proliferation.
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- 2017
48. MiR-675-5p supports hypoxia induced epithelial to mesenchymal transition in colon cancer cells
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Marco Tripodi, Viviana Costa, Alessia Lo Dico, Alice Conigliaro, Francesca Rajata, Aroldo Rizzo, Riccardo Alessandro, Costa, V., Lo Dico, A., Rizzo, A., Rajata, F., Tripodi, M., Alessandro, R., and Conigliaro, A.
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0301 basic medicine ,Pathology ,medicine.medical_specialty ,miRNA675 ,Epithelial-Mesenchymal Transition ,Transcription, Genetic ,Colorectal cancer ,Down-Regulation ,Metastasi ,Metastasis ,03 medical and health sciences ,0302 clinical medicine ,Glioma ,Cell Line, Tumor ,medicine ,metastasis ,Humans ,Epithelial–mesenchymal transition ,Neoplasm Metastasis ,Lymph node ,Metastatic colon cancer ,CRC ,EMT ,Hypoxia ,MiRNA675 ,Oncology ,business.industry ,hypoxia ,Hypoxia (medical) ,medicine.disease ,Hypoxia-Inducible Factor 1, alpha Subunit ,Cell Hypoxia ,Transplantation ,DNA-Binding Proteins ,MicroRNAs ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Colonic Neoplasms ,Cancer research ,medicine.symptom ,business ,Research Paper - Abstract
// Viviana Costa 1, * , Alessia Lo Dico 2, * , Aroldo Rizzo 3 , Francesca Rajata 3 , Marco Tripodi 4, 5 , Riccardo Alessandro 6, 7, * , Alice Conigliaro 4, * 1 Innovative Technological Platforms for Tissue Engineering, Theranostic and Oncology, Rizzoli Orthopedic Institute, Palermo, Italy 2 Department of Pathophysiology and Transplantation, Universita degli Studi di Milano, Milano, Italy 3 Unita Operativa di Anatomia Patologica, Azienda Ospedaliera Ospedali Riuniti “Villa Sofia-Cervello”, Palermo, Italy 4 Dipartimento di Biotecnologie Cellulari ed Ematologia, Sapienza University of Rome, Rome, Italy 5 National Institute for Infectious Diseases L. Spallanzani, IRCCS, Rome, Italy 6 Dipartimento di Biopatologia e Biotecnologie Mediche, University of Palermo, Palermo, Italy 7 Institute of Biomedicine and Molecular Immunology (IBIM), National Research Council of Italy, Palermo, Italy * These authors contributed equally to this work Correspondence to: Alice Conigliaro, email: conigliaro@bce.uniroma1.it Riccardo Alessandro, email: riccardo.alessandro@unipa.it Keywords: miRNA675, CRC, EMT, metastasis, hypoxia Received: November 16, 2016 Accepted: December 27, 2016 Published: January 03, 2017 ABSTRACT The survival rates in colon cancer patients are inversely proportional to the number of lymph node metastases. The hypoxia-induced Epithelial to Mesenchymal Transition (EMT), driven by HIF1α, is known to be involved in cancer progression and metastasis. Recently, we have reported that miR-675-5p promotes glioma growth by stabilizing HIF1α; here, by use of the syngeneic cell lines we investigated the role of the miR-675-5p in colon cancer metastasis. Our results show that miR-675-5p , over expressed in metastatic colon cancer cells, participates to tumour progression by regulating HIF1α induced EMT. MiR-675-5p increases Snail transcription by a dual strategy: i) stabilizing the activity of the transcription factor HIF1α and ii) and inhibiting Snail’s repressor DDB2 (Damage specific DNA Binding protein 2). Moreover, transcriptional analyses on specimens from colon cancer patients confirmed, in vivo , the correlation between miR-675-5p over-expression and metastasis, thus identifying miR-675-5p as a new marker for colon cancer progression and therefore a putative target for therapeutic strategies.
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- 2017
49. Molecular mechanisms controlling the phenotype and the EMT/MET dynamics of hepatocyte
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Marco Tripodi, Carmine Mancone, Tonino Alonzi, Carla Cicchini, Alessandra Marchetti, Laura Amicone, Department of Cellular Biotechnologies and Haematology, Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome]-Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), L. Spallanzani National Institute for Infectious Diseases, and IRCCS
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EMT ,Snai ,l, HNF4alpha ,hepatocytes ,Epithelial-Mesenchymal Transition ,Cellular differentiation ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Biology ,EMT/MET ,HNF4a ,Models, Biological ,03 medical and health sciences ,Paracrine signalling ,0302 clinical medicine ,Humans ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Hepatocyte ,Epithelial–mesenchymal transition ,Review Articles ,030304 developmental biology ,Regulation of gene expression ,Hepatocyte differentiation ,Genetics ,0303 health sciences ,Hepatology ,Phenotype ,Cell biology ,MicroRNAs ,Hepatocyte nuclear factor 4 ,Snail ,Gene Expression Regulation ,Hepatocyte Nuclear Factor 4 ,030220 oncology & carcinogenesis ,Hepatocytes ,Snail Family Transcription Factors ,Stem cell ,Transcription Factors - Abstract
International audience; The complex spatial and paracrine relationships between the various liver histotypes are essential for proper functioning of the hepatic parenchymal cells. Only within a correct tissue organization, in fact, they stably maintain their identity and differentiated phenotype. The loss of histotype identity, which invariably occurs in the primary hepatocytes in culture, or in vivo in particular pathological conditions (fibrosis and tumours), is mainly because of the phenomenon of epithelial-to-mesenchymal transition (EMT). The EMT process, that occurs in the many epithelial cells, appears to be driven by a number of general, non-tissue-specific, master transcriptional regulators. The reverse process, the mesenchymal-to-epithelial transition (MET), as yet much less characterized at a molecular level, restores specific epithelial identities, and thus must include tissue-specific master elements. In this review, we will summarize the so far unveiled events of EMT/MET occurring in liver cells. In particular, we will focus on hepatocyte and describe the pivotal role in the control of EMT/MET dynamics exerted by a tissue-specific molecular mini-circuitry. Recent evidence, indeed, highlighted as two transcriptional factors, the master gene of EMT Snail, and the master gene of hepatocyte differentiation HNF4α, exhorting a direct reciprocal repression, act as pivotal elements in determining opposite cellular outcomes. The different balances between these two master regulators, further integrated by specific microRNAs, in fact, were found responsible for the EMT/METs dynamics as well as for the preservation of both hepatocyte and stem/precursor cells identity and differentiation. Overall, these findings impact the maintenance of stem cells and differentiated cells both in in vivo EMT/MET physio-pathological processes as well as in culture.
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- 2014
50. Spike-in SILAC proteomic approach reveals the vitronectin as an early molecular signature of liver fibrosis in hepatitis C infections with hepatic iron overload
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Leopoldo Paolo Pucillo, Andrea Baiocchini, Carmine Mancone, Franca Del Nonno, Marco Tripodi, Vera van Noort, Tonino Alonzi, Claudia Montaldo, Nicolina Rotiroti, Giuseppe Ippolito, Laura Amicone, Alice Conigliaro, Angela Maria Cozzolino, Cecilia Battistelli, Simone Mattei, Montaldo, C., Mattei, S., Baiocchini, A., Rotiroti, N., Nonno, F., Pucillo, L., Cozzolino, A., Battistelli, C., Amicone, L., Ippolito, G., van Noort, V., Conigliaro, A., Alonzi, T., Tripodi, M., Mancone, C., Department of Cellular Biotechnologies and Haematology, Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome]-Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), L. Spallanzani, National Institute for Infectious Diseases (IRCCS), European Molecular Biology Laboratory [Heidelberg] (EMBL), and This work was supported by grants from MIUR Ministero dell’Universit `a e Ricerca Scientifica (FIRB 2012, codice progetto RBFR12NSCF), Associazione Italiana per la Ricerca sul Cancro (AIRC) andMinistero della Salute (Ricerca Finalizzata 40H27, Ricerca Corrente).
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Liver Cirrhosis ,Proteomics ,hepatitis C virus ,Male ,MESH: Isotope Labeling ,HSC ,medicine.disease_cause ,Biochemistry ,0302 clinical medicine ,Fibrosis ,MESH: Up-Regulation ,Membrane Protein ,hepatic stellate cell ,liver fibrosis ,hepatic iron overload ,0303 health sciences ,biology ,MESH: Proteomics ,Medicine (all) ,hepatocellular carcinoma ,Biomedicine ,hepatitis c infection ,vitronectin ,Hepatitis C ,[SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM] ,Up-Regulation ,3. Good health ,cell culture-derived HCV ,Isotope Labeling ,030220 oncology & carcinogenesis ,Hepatocellular carcinoma ,Hepatic iron overload ,Hepatitis C infection ,Liver fibrosis ,Vitronectin ,Biomarkers ,Cell Line ,Humans ,Iron Overload ,Membrane Proteins ,Molecular Biology ,HCV ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,Biomarker (medicine) ,MESH: Membrane Proteins ,MESH: Liver Cirrhosis ,Human ,Liver Cirrhosi ,Hepatitis C virus ,MESH: Iron Overload ,03 medical and health sciences ,medicine ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,030304 developmental biology ,MESH: Hepatitis C ,MESH: Humans ,MESH: Biological Markers ,Liver fibrosi ,Proteomic ,Biomarker ,medicine.disease ,MESH: Vitronectin ,MESH: Male ,digestive system diseases ,MESH: Cell Line ,Biomedicine / Abbreviations: HCC ,HCVcc ,Immunology ,Cancer research ,Hepatic stellate cell ,biology.protein ,Steatosis - Abstract
Hepatitis C virus (HCV)-induced iron overload has been shown to promote liver fibrosis, steatosis, and hepatocellular carcinoma. The zonal-restricted histological distribution of pathological iron deposits has hampered the attempt to perform large-scale in vivo molecular investigations on the comorbidity between iron and HCV. Diagnostic and prognostic markers are not yet available to assess iron overload-induced liver fibrogenesis and progression in HCV infections. Here, by means of Spike-in SILAC proteomic approach, we first unveiled a specific membrane protein expression signature of HCV cell cultures in the presence of iron overload. Computational analysis of proteomic dataset highlighted the hepatocytic vitronectin expression as the most promising specific biomarker for iron-associated fibrogenesis in HCV infections. Next, the robustness of our in vitro findings was challenged in human liver biopsies by immunohistochemistry and yielded two major results: (i) hepatocytic vitronectin expression is associated to liver fibrogenesis in HCV-infected patients with iron overload; (ii) hepatic vitronectin expression was found to discriminate also the transition between mild to moderate fibrosis in HCV-infected patients without iron overload. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
- Published
- 2014
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