A Romeo A. Desideri Cesare E.M. Gruber D. Pietrucci F. Cardamone F. Iacovelli M. Falconi S Biocca S Raniolo
2018 |
Journal Articles |
Menduti, G; Biamino, E; Vittorini, R; Vesco, S; Puccinelli, MP; Porta, F; Capo, C; Leo, S; Ciminelli, BM; Iacovelli, F; Spada, M; Falconi, M; Malaspina, P; Rossi, L Molecular Genetics and Metabolism, 124 , pp. 210-215, 2018. Abstract | Links | BibTeX | Tags: F. Iacovelli, M. Falconi @article{Menduti2008, title = {Succinic semialdehyde dehydrogenase deficiency: The combination of a novel ALDH5A1 gene mutation and a missense SNP strongly affects SSADH enzyme activity and stability}, author = {G Menduti and E Biamino and R Vittorini and S Vesco and MP Puccinelli and F Porta and C Capo and S Leo and BM Ciminelli and F Iacovelli and M Spada and M Falconi and P Malaspina and L Rossi}, doi = {10.1016/j.ymgme.2018.05.006}, year = {2018}, date = {2018-06-02}, journal = {Molecular Genetics and Metabolism}, volume = {124}, pages = {210-215}, abstract = {Succinic semialdehyde dehydrogenase deficiency (SSADHD) is a rare autosomal recessive metabolic disorder of GABA catabolism. SSADH is a mitochondrial homotetrameric enzyme encoded by ALDH5A1 gene. We report the molecular characterization of ALDH5A1 gene in an Italian SSADHD patient, showing heterozygosity for four missense mutations: c.526G>A (p.G176R), c.538C>T (p.H180Y), c.709G>T (p.A237S) and c.1267A>T (p.T423S), the latter never described so far. The patient inherited c.526A in cis with c.538T from the mother and c.709T in cis with c.1267T from the father. To explore the effects of the two allelic arrangements on SSADH activity and protein level, wild type, single or double mutated cDNA constructs were expressed in a cell system. The p.G176R change, alone or in combination with p.H180Y, causes the abolishment of enzyme activity. Western blot analysis showed a strongly reduced amount of the p.176R-p.180Y double mutant protein, suggesting increased degradation. Indeed, in silico analyses confirmed high instability of this mutant homotetramer. Enzyme activity relative to the other p.423S-p.237S double mutant is around 30% of wt. Further in silico analyses on all the possible combinations of mutant monomers suggest the lowest stability for the tetramer constituted by p.176R-p.180Y monomers and the highest stability for that constituted by p.237S-p.423S monomers. The present study shows that when a common SNP, associated with a slight reduction of SSADH activity, is inherited in cis with a mutation showing no consequences on the enzyme function, the activity is strongly affected. In conclusion, the peculiar arrangement of four missense mutations occurring in this patient is responsible for the SSADHD phenotype.}, keywords = {F. Iacovelli, M. Falconi}, pubstate = {published}, tppubtype = {article} } Succinic semialdehyde dehydrogenase deficiency (SSADHD) is a rare autosomal recessive metabolic disorder of GABA catabolism. SSADH is a mitochondrial homotetrameric enzyme encoded by ALDH5A1 gene. We report the molecular characterization of ALDH5A1 gene in an Italian SSADHD patient, showing heterozygosity for four missense mutations: c.526G>A (p.G176R), c.538C>T (p.H180Y), c.709G>T (p.A237S) and c.1267A>T (p.T423S), the latter never described so far. The patient inherited c.526A in cis with c.538T from the mother and c.709T in cis with c.1267T from the father. To explore the effects of the two allelic arrangements on SSADH activity and protein level, wild type, single or double mutated cDNA constructs were expressed in a cell system. The p.G176R change, alone or in combination with p.H180Y, causes the abolishment of enzyme activity. Western blot analysis showed a strongly reduced amount of the p.176R-p.180Y double mutant protein, suggesting increased degradation. Indeed, in silico analyses confirmed high instability of this mutant homotetramer. Enzyme activity relative to the other p.423S-p.237S double mutant is around 30% of wt. Further in silico analyses on all the possible combinations of mutant monomers suggest the lowest stability for the tetramer constituted by p.176R-p.180Y monomers and the highest stability for that constituted by p.237S-p.423S monomers. The present study shows that when a common SNP, associated with a slight reduction of SSADH activity, is inherited in cis with a mutation showing no consequences on the enzyme function, the activity is strongly affected. In conclusion, the peculiar arrangement of four missense mutations occurring in this patient is responsible for the SSADHD phenotype. |
Errico, V; Arrabito, G; Fornetti, E; Fuoco, C; Testa, S; Saggio, G; Rufini, S; Cannata, S; Desideri, A; Falconi, C; Gargioli, C High-Density ZnO Nanowires as a Reversible Myogenic-Differentiation Switch Journal Article ACS Applied Materials and Interfaces, 10 , pp. 14097-14107, 2018. Abstract | Links | BibTeX | Tags: A. Desideri @article{Errico2018, title = {High-Density ZnO Nanowires as a Reversible Myogenic-Differentiation Switch}, author = {V Errico and G Arrabito and E Fornetti and C Fuoco and S Testa and G Saggio and S Rufini and S Cannata and A Desideri and C Falconi and C Gargioli }, doi = {10.1021/acsami.7b19758}, year = {2018}, date = {2018-04-25}, journal = {ACS Applied Materials and Interfaces}, volume = {10}, pages = {14097-14107}, abstract = {Mesoangioblasts are outstanding candidates for stem-cell therapy and are already being explored in clinical trials. However, a crucial challenge in regenerative medicine is the limited availability of undifferentiated myogenic progenitor cells because growth is typically accompanied by differentiation. Here reversible myogenic-differentiation switching during proliferation is achieved by functionalizing the glass substrate with high-density ZnO nanowires (NWs). Specifically, mesoangioblasts grown on ZnO NWs present a spherical viable undifferentiated cell state without lamellopodia formation during the entire observation time (8 days). Consistently, the myosin heavy chain, typically expressed in skeletal muscle tissue and differentiated myogenic progenitors, is completely absent. Remarkably, NWs do not induce any damage while they reversibly block differentiation, so that the differentiation capabilities are completely recovered upon cell removal from the NW-functionalized substrate and replating on standard culture glass. This is the first evidence of a reversible myogenic-differentiation switch that does not affect the viability. These results can be the first step toward for the in vitro growth of a large number of undifferentiated stem/progenitor cells and therefore can represent a breakthrough for cell-based therapy and tissue engineering.}, keywords = {A. Desideri}, pubstate = {published}, tppubtype = {article} } Mesoangioblasts are outstanding candidates for stem-cell therapy and are already being explored in clinical trials. However, a crucial challenge in regenerative medicine is the limited availability of undifferentiated myogenic progenitor cells because growth is typically accompanied by differentiation. Here reversible myogenic-differentiation switching during proliferation is achieved by functionalizing the glass substrate with high-density ZnO nanowires (NWs). Specifically, mesoangioblasts grown on ZnO NWs present a spherical viable undifferentiated cell state without lamellopodia formation during the entire observation time (8 days). Consistently, the myosin heavy chain, typically expressed in skeletal muscle tissue and differentiated myogenic progenitors, is completely absent. Remarkably, NWs do not induce any damage while they reversibly block differentiation, so that the differentiation capabilities are completely recovered upon cell removal from the NW-functionalized substrate and replating on standard culture glass. This is the first evidence of a reversible myogenic-differentiation switch that does not affect the viability. These results can be the first step toward for the in vitro growth of a large number of undifferentiated stem/progenitor cells and therefore can represent a breakthrough for cell-based therapy and tissue engineering. |
Mastrorilli, E; Pietrucci, D; Barco, L; Ammendola, S; Petrin, S; Longo, A; Mantovani, C; Battistoni, A; Ricci, A; Desideri, A; Losasso, C A comparative genomic analysis provides novel insights into the ecological success of the monophasic Salmonella serovar 4,[5],12:i:- Journal Article Frontiers in Microbiology, 9 , pp. 715, 2018. Abstract | Links | BibTeX | Tags: A. Desideri, D. Pietrucci @article{Mastrorilli2018, title = {A comparative genomic analysis provides novel insights into the ecological success of the monophasic Salmonella serovar 4,[5],12:i:-}, author = {E Mastrorilli and D Pietrucci and L Barco and S Ammendola and S Petrin and A Longo and C Mantovani and A Battistoni and A Ricci and A Desideri and C Losasso}, doi = {10.3389/fmicb.2018.00715}, year = {2018}, date = {2018-04-17}, journal = {Frontiers in Microbiology}, volume = {9}, pages = {715}, abstract = {Over the past decades, Salmonella 4,[5],12:i:- has rapidly emerged and it is isolated with high frequency in the swine food chain. Although many studies have documented the epidemiological success of this serovar, few investigations have tried to explain this phenomenon from a genetic perspective. Here a comparative whole-genome analysis of 50 epidemiologically unrelated S. 4,[5],12:i:-, isolated in Italy from 2010 to 2016 was performed, characterizing them in terms of genetic elements potentially conferring resistance, tolerance and persistence characteristics. Phylogenetic analyses indicated interesting distinctions among the investigated isolates. The most striking genetic trait characterizing the analyzed isolates is the widespread presence of heavy metals tolerance gene cassettes: most of the strains possess genes expected to confer resistance to copper and silver, whereas about half of the isolates also contain the mercury tolerance gene merA. A functional assay showed that these genes might be useful for preventing the toxic effects of metals, thus supporting the hypothesis that they can contribute to the success of S. 4,[5],12:i:- in farming environments. In addition, the analysis of the distribution of type II toxin-antitoxin families indicated that these elements are abundant in this serovar, suggesting that this is another factor that might favor its successful spread. }, keywords = {A. Desideri, D. Pietrucci}, pubstate = {published}, tppubtype = {article} } Over the past decades, Salmonella 4,[5],12:i:- has rapidly emerged and it is isolated with high frequency in the swine food chain. Although many studies have documented the epidemiological success of this serovar, few investigations have tried to explain this phenomenon from a genetic perspective. Here a comparative whole-genome analysis of 50 epidemiologically unrelated S. 4,[5],12:i:-, isolated in Italy from 2010 to 2016 was performed, characterizing them in terms of genetic elements potentially conferring resistance, tolerance and persistence characteristics. Phylogenetic analyses indicated interesting distinctions among the investigated isolates. The most striking genetic trait characterizing the analyzed isolates is the widespread presence of heavy metals tolerance gene cassettes: most of the strains possess genes expected to confer resistance to copper and silver, whereas about half of the isolates also contain the mercury tolerance gene merA. A functional assay showed that these genes might be useful for preventing the toxic effects of metals, thus supporting the hypothesis that they can contribute to the success of S. 4,[5],12:i:- in farming environments. In addition, the analysis of the distribution of type II toxin-antitoxin families indicated that these elements are abundant in this serovar, suggesting that this is another factor that might favor its successful spread. |
Cardamone, F; Falconi, M; Desideri, A Molecular dynamics characterization of the SAMHD1 Aicardi–Goutières Arg145Gln mutant: structural determinants for the impaired tetramerization Journal Article Journal of Computer-Aided Molecular Design, 32 , pp. 623-632, 2018. Abstract | BibTeX | Tags: A. Desideri, F. Cardamone, M. Falconi @article{Cardamone2018, title = {Molecular dynamics characterization of the SAMHD1 Aicardi–Goutières Arg145Gln mutant: structural determinants for the impaired tetramerization}, author = {F Cardamone and M Falconi and A Desideri}, year = {2018}, date = {2018-03-28}, journal = {Journal of Computer-Aided Molecular Design}, volume = {32}, pages = {623-632}, abstract = {Aicardi–Goutières syndrome, a rare genetic disorder characterized by calcification of basal ganglia, results in psychomotor delays and epilepsy states from the early months of children life. This disease is caused by mutations in seven different genes encoding proteins implicated in the metabolism of nucleic acids, including SAMHD1. Twenty SAMHD1 gene variants have been discovered and in this work, a structural characterization of the SAMHD1 Aicardi–Goutières Arg145Gln mutant is reported by classical molecular dynamics simulation. Four simulations have been carried out and compared. Two concerning the wild-type SAMHD1 form in presence and absence of cofactors, in order to explain the role of cofactors in the SAMHD1 assembly/disassembly process and, two concerning the Arg145Gln mutant, also in presence and absence of cofactors, in order to have an accurate comparison with the corresponding native forms. Results show the importance of native residue Arg145 in maintaining the tetramer, interacting with GTP cofactor inside allosteric sites. Replacement of arginine in glutamine gives rise to a loosening of GTP–protein interactions, when cofactors are present in allosteric sites, whilst in absence of cofactors, the occurrence of intra and inter-chain interactions is observed in the mutant, not seen in the native enzyme, making energetically unfavourable the tetramerization process}, keywords = {A. Desideri, F. Cardamone, M. Falconi}, pubstate = {published}, tppubtype = {article} } Aicardi–Goutières syndrome, a rare genetic disorder characterized by calcification of basal ganglia, results in psychomotor delays and epilepsy states from the early months of children life. This disease is caused by mutations in seven different genes encoding proteins implicated in the metabolism of nucleic acids, including SAMHD1. Twenty SAMHD1 gene variants have been discovered and in this work, a structural characterization of the SAMHD1 Aicardi–Goutières Arg145Gln mutant is reported by classical molecular dynamics simulation. Four simulations have been carried out and compared. Two concerning the wild-type SAMHD1 form in presence and absence of cofactors, in order to explain the role of cofactors in the SAMHD1 assembly/disassembly process and, two concerning the Arg145Gln mutant, also in presence and absence of cofactors, in order to have an accurate comparison with the corresponding native forms. Results show the importance of native residue Arg145 in maintaining the tetramer, interacting with GTP cofactor inside allosteric sites. Replacement of arginine in glutamine gives rise to a loosening of GTP–protein interactions, when cofactors are present in allosteric sites, whilst in absence of cofactors, the occurrence of intra and inter-chain interactions is observed in the mutant, not seen in the native enzyme, making energetically unfavourable the tetramerization process |
Raniolo, S; Vindigni, G; Ottaviani, A; Unida, V; Iacovelli, F; Manetto, A; Figini, M; Stella, L; Desideri, A; Biocca, S Selective targeting and degradation of doxorubicin-loaded folate-functionalized DNA nanocages. Journal Article Nanomedicine, 14 , pp. 1181-1190, 2018. Abstract | Links | BibTeX | Tags: A. Desideri, F. Iacovelli, S Biocca, S Raniolo @article{Raniolo20180216, title = {Selective targeting and degradation of doxorubicin-loaded folate-functionalized DNA nanocages.}, author = {S Raniolo and G Vindigni and A Ottaviani and V Unida and F Iacovelli and A Manetto and M Figini and L Stella and A Desideri and S Biocca}, url = {https://www.sciencedirect.com/science/article/pii/S1549963418300340?via%3Dihub}, doi = {10.1016/j.nano.2018.02.002}, year = {2018}, date = {2018-02-17}, journal = {Nanomedicine}, volume = {14}, pages = {1181-1190}, abstract = {Selective targeting is a crucial property of nanocarriers used for drug delivery in cancer therapy. We generated biotinylated octahedral DNA nanocages functionalized with folic acid through bio-orthogonal conjugation chemistry. Molecular modelling indicated that a distance of about 2.5 nm between folic acid and DNA nanocage avoids steric hindrance with the folate receptor. HeLa cells, a folate receptor positive tumour cell line, internalize folate-DNA nanocages with efficiency greater than 40 times compared to cells not expressing the folate receptors. Functionalized DNA nanocages are highly stable, not cytotoxic and can be efficiently loaded with the chemotherapeutic agent doxorubicin. After entry into cells, doxorubicin-loaded nanoparticles are confined in vesicular structures, indicating that DNA nanocages traffic through the endocytic pathway. Doxorubicin release from loaded DNA cages, facilitated by low pH of endocytic vesicles, induces toxic pathways that, besides selectively killing folate receptor-positive cancer cells, lead to cage degradation avoiding nanoparticles accumulation inside cells.}, keywords = {A. Desideri, F. Iacovelli, S Biocca, S Raniolo}, pubstate = {published}, tppubtype = {article} } Selective targeting is a crucial property of nanocarriers used for drug delivery in cancer therapy. We generated biotinylated octahedral DNA nanocages functionalized with folic acid through bio-orthogonal conjugation chemistry. Molecular modelling indicated that a distance of about 2.5 nm between folic acid and DNA nanocage avoids steric hindrance with the folate receptor. HeLa cells, a folate receptor positive tumour cell line, internalize folate-DNA nanocages with efficiency greater than 40 times compared to cells not expressing the folate receptors. Functionalized DNA nanocages are highly stable, not cytotoxic and can be efficiently loaded with the chemotherapeutic agent doxorubicin. After entry into cells, doxorubicin-loaded nanoparticles are confined in vesicular structures, indicating that DNA nanocages traffic through the endocytic pathway. Doxorubicin release from loaded DNA cages, facilitated by low pH of endocytic vesicles, induces toxic pathways that, besides selectively killing folate receptor-positive cancer cells, lead to cage degradation avoiding nanoparticles accumulation inside cells. |
Wang, Z; Ouyang, H; Tesauro, C; Ottaviani, A; He, Y; Fiorani, P; Xie, H; Desideri, A; Fu, Z 643, pp. 1-6, 2018. Abstract | Links | BibTeX | Tags: A. Desideri @article{Wang2018b, title = {Real-time analysis of cleavage and religation activity of human topoisomerase 1 based on ternary fluorescence resonance energy transfer DNA substrate}, author = {Z Wang and H Ouyang and C Tesauro and A Ottaviani and Y He and P Fiorani and H Xie and A Desideri and Z Fu}, doi = {10.1016/j.abb.2018.02.006}, year = {2018}, date = {2018-02-16}, journal = {643}, pages = {1-6}, abstract = {Human topoisomerase 1B is a ubiquitous and essential enzyme involved in relaxing the topological state of supercoiled DNA to allow the progression of fundamental DNA metabolism. Its enzymatic catalytic cycle consists of cleavage and religation reaction. A ternary fluorescence resonance energy transfer biosensor based on a suicide DNA substrate conjugated with three fluorophores has been developed to monitor both cleavage and religation Topoisomerase I catalytic function. The presence of fluorophores does not alter the specificity of the enzyme catalysis on the DNA substrate. The enzyme-mediated reaction can be tracked in real-time by simple fluorescence measurement, avoiding the use of risky radioactive substrate labeling and time-consuming denaturing gel electrophoresis. The method is applied to monitor the perturbation brought by single mutation on the cleavage or religation reaction and to screen the effect of the camptothecin anticancer drug monitoring the energy transfer decrease during religation reaction. Pathological mutations usually affect only the cleavage or the religation reaction and the proposed approach represent a fast protocol for assessing chemotherapeutic drug efficacy and analyzing mutant's properties. }, keywords = {A. Desideri}, pubstate = {published}, tppubtype = {article} } Human topoisomerase 1B is a ubiquitous and essential enzyme involved in relaxing the topological state of supercoiled DNA to allow the progression of fundamental DNA metabolism. Its enzymatic catalytic cycle consists of cleavage and religation reaction. A ternary fluorescence resonance energy transfer biosensor based on a suicide DNA substrate conjugated with three fluorophores has been developed to monitor both cleavage and religation Topoisomerase I catalytic function. The presence of fluorophores does not alter the specificity of the enzyme catalysis on the DNA substrate. The enzyme-mediated reaction can be tracked in real-time by simple fluorescence measurement, avoiding the use of risky radioactive substrate labeling and time-consuming denaturing gel electrophoresis. The method is applied to monitor the perturbation brought by single mutation on the cleavage or religation reaction and to screen the effect of the camptothecin anticancer drug monitoring the energy transfer decrease during religation reaction. Pathological mutations usually affect only the cleavage or the religation reaction and the proposed approach represent a fast protocol for assessing chemotherapeutic drug efficacy and analyzing mutant's properties. |
2017 |
Journal Articles |
Ferese, R; Albano, V; Falconi, M; Iacovelli, F; Campopiano, R; Scala, S; Griguoli, AM; Gaglione, A; Giardina, E; Zampatti, S; Storto, M; Fornai, F; D'Alessio, C; Novelli, G; Gambardella, S Structural modeling of altered CLCN1 conformation following a novel mutation in a patient affected by autosomal dominant myotonia congenita (Thomsen disease). Journal Article Forthcoming Archives italiennes de biologie, 155 (4), pp. 118, Forthcoming. Links | BibTeX | Tags: F. Iacovelli, M. Falconi @article{Ferese20171201, title = {Structural modeling of altered CLCN1 conformation following a novel mutation in a patient affected by autosomal dominant myotonia congenita (Thomsen disease).}, author = {R Ferese and V Albano and M Falconi and F Iacovelli and R Campopiano and S Scala and AM Griguoli and A Gaglione and E Giardina and S Zampatti and M Storto and F Fornai and C D'Alessio and G Novelli and S Gambardella}, doi = {10.12871/000398292017410}, year = {2017}, date = {2017-12-01}, journal = {Archives italiennes de biologie}, volume = {155}, number = {4}, pages = {118}, keywords = {F. Iacovelli, M. Falconi}, pubstate = {forthcoming}, tppubtype = {article} } |
Takarada, J E; Guedes, A P M; Correa, R S; Silveira-Lacerda, E D P; Castelli, S; Iacovelli, F; Deflon, V M; Batista, A A; Desideri, A Ru/Fe bimetallic complexes: Synthesis, characterization, cytotoxicity and study of their interactions with DNA/HSA and human topoisomerase IB Journal Article Archives of Biochemistry and Biophysics, 636 , pp. 28-41, 2017, (cited By 0). Links | BibTeX | Tags: A. Desideri, F. Iacovelli @article{Takarada201728, title = {Ru/Fe bimetallic complexes: Synthesis, characterization, cytotoxicity and study of their interactions with DNA/HSA and human topoisomerase IB}, author = {J E Takarada and A P M Guedes and R S Correa and E D P Silveira-Lacerda and S Castelli and F Iacovelli and V M Deflon and A A Batista and A Desideri}, url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85033409950&doi=10.1016%2fj.abb.2017.10.015&partnerID=40&md5=b39e375ebb19a38efa2f0f3ab1b43c7e}, doi = {10.1016/j.abb.2017.10.015}, year = {2017}, date = {2017-01-01}, journal = {Archives of Biochemistry and Biophysics}, volume = {636}, pages = {28-41}, note = {cited By 0}, keywords = {A. Desideri, F. Iacovelli}, pubstate = {published}, tppubtype = {article} } |
Capo, C R; Pedersen, J Z; Falconi, M; Rossi, L Oleuropein shows copper complexing properties and noxious effect on cultured SH-SY5Y neuroblastoma cells depending on cell copper content Journal Article Journal of Trace Elements in Medicine and Biology, 44 , pp. 225-232, 2017, (cited By 0). Links | BibTeX | Tags: M. Falconi @article{Capo2017225, title = {Oleuropein shows copper complexing properties and noxious effect on cultured SH-SY5Y neuroblastoma cells depending on cell copper content}, author = {C R Capo and J Z Pedersen and M Falconi and L Rossi}, url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85027515292&doi=10.1016%2fj.jtemb.2017.08.002&partnerID=40&md5=42e79a7aa54eb67cd3a93c204bdfab3a}, doi = {10.1016/j.jtemb.2017.08.002}, year = {2017}, date = {2017-01-01}, journal = {Journal of Trace Elements in Medicine and Biology}, volume = {44}, pages = {225-232}, note = {cited By 0}, keywords = {M. Falconi}, pubstate = {published}, tppubtype = {article} } |
Iacovelli, F; Tucci, F G; Macari, G; Falconi, M Proteins: Structure, Function and Bioinformatics, 85 (10), pp. 1902-1912, 2017, (cited By 0). Links | BibTeX | Tags: F. Iacovelli, M. Falconi @article{Iacovelli20171902, title = {Multiple molecular dynamics simulations of human LOX-1 and Trp150Ala mutant reveal the structural determinants causing the full deactivation of the receptor}, author = {F Iacovelli and F G Tucci and G Macari and M Falconi}, url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85023184186&doi=10.1002%2fprot.25344&partnerID=40&md5=7d12acf2688df26d58034750eddc1b3e}, doi = {10.1002/prot.25344}, year = {2017}, date = {2017-01-01}, journal = {Proteins: Structure, Function and Bioinformatics}, volume = {85}, number = {10}, pages = {1902-1912}, note = {cited By 0}, keywords = {F. Iacovelli, M. Falconi}, pubstate = {published}, tppubtype = {article} } |