Kawasumi R; Abe T; Arakawa H; Garre M; Hirota K; Branzei D ESCO1/2's roles in chromosome structure and interphase chromatin organization. Journal Article In: Genes and development, vol. 31, no 21, pp. 2136-2150, 2017. @article{%a1:%Y_192,
title = {ESCO1/2's roles in chromosome structure and interphase chromatin organization.},
author = {Kawasumi R and Abe T and Arakawa H and Garre M and Hirota K and Branzei D},
url = {http://genesdev.cshlp.org/content/31/21/2136.long},
doi = {10.1101/gad.306084.117},
year = {2017},
date = {2017-08-10},
journal = {Genes and development},
volume = {31},
number = {21},
pages = {2136-2150},
abstract = {ESCO1/2 acetyltransferases mediating SMC3 acetylation and sister chromatid cohesion (SCC) are differentially required for genome integrity and development. Here we established chicken DT40 cell lines with mutations in ESCO1/2, SMC3 acetylation, and the cohesin remover WAPL. Both ESCO1 and ESCO2 promoted SCC, while ESCO2 was additionally and specifically required for proliferation and centromere integrity. ESCO1 overexpression fully suppressed the slow proliferation and centromeric separation phenotypes of esco2 cells but only partly suppressed its chromosome arm SCC defects. Concomitant inactivation of ESCO1 and ESCO2 caused lethality owing to compromised mitotic chromosome segregation. Neither wapl nor acetyl-mimicking smc3-QQ mutations rescued esco1 esco2 lethality. Notably, esco1 esco2 wapl conditional mutants showed very severe proliferation defects associated with catastrophic mitoses and also abnormal interphase chromatin organization patterns. The results indicate that cohesion establishment by vertebrate ESCO1/2 is linked to interphase chromatin architecture formation, a newly identified function of cohesin acetyltransferases that is both fundamentally and medically relevant.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
ESCO1/2 acetyltransferases mediating SMC3 acetylation and sister chromatid cohesion (SCC) are differentially required for genome integrity and development. Here we established chicken DT40 cell lines with mutations in ESCO1/2, SMC3 acetylation, and the cohesin remover WAPL. Both ESCO1 and ESCO2 promoted SCC, while ESCO2 was additionally and specifically required for proliferation and centromere integrity. ESCO1 overexpression fully suppressed the slow proliferation and centromeric separation phenotypes of esco2 cells but only partly suppressed its chromosome arm SCC defects. Concomitant inactivation of ESCO1 and ESCO2 caused lethality owing to compromised mitotic chromosome segregation. Neither wapl nor acetyl-mimicking smc3-QQ mutations rescued esco1 esco2 lethality. Notably, esco1 esco2 wapl conditional mutants showed very severe proliferation defects associated with catastrophic mitoses and also abnormal interphase chromatin organization patterns. The results indicate that cohesion establishment by vertebrate ESCO1/2 is linked to interphase chromatin architecture formation, a newly identified function of cohesin acetyltransferases that is both fundamentally and medically relevant. |
Rohban S; Cerutti A; Morelli MJ; d'Adda di Fagagna F; Campaner S The cohesin complex prevents Myc-induced replication stress. Journal Article In: Cell Death and Disease, vol. 8, pp. e2956, 2017. @article{%a1:%Y_201,
title = {The cohesin complex prevents Myc-induced replication stress.},
author = {Rohban S and Cerutti A and Morelli MJ and {d'Adda di Fagagna F} and Campaner S},
url = {https://www.nature.com/articles/cddis2017345},
doi = {10.1038/cddis.2017.345},
year = {2017},
date = {2017-07-27},
journal = {Cell Death and Disease},
volume = {8},
pages = {e2956},
abstract = {The cohesin complex is mutated in cancer and in a number of rare syndromes collectively known as Cohesinopathies. In the latter case, cohesin deficiencies have been linked to transcriptional alterations affecting Myc and its target genes. Here, we set out to understand to what extent the role of cohesins in controlling cell cycle is dependent on Myc expression and activity. Inactivation of the cohesin complex by silencing the RAD21 subunit led to cell cycle arrest due to both transcriptional impairment of Myc target genes and alterations of replication forks, which were fewer and preferentially unidirectional. Ectopic activation of Myc in RAD21 depleted cells rescued Myc-dependent transcription and promoted S-phase entry but failed to sustain S-phase progression due to a strong replicative stress response, which was associated to a robust DNA damage response, DNA damage checkpoint activation and synthetic lethality. Thus, the cohesin complex is dispensable for Myc-dependent transcription but essential to prevent Myc-induced replicative stress. This suggests the presence of a feed-forward regulatory loop where cohesins by regulating Myc level control S-phase entry and prevent replicative stress.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The cohesin complex is mutated in cancer and in a number of rare syndromes collectively known as Cohesinopathies. In the latter case, cohesin deficiencies have been linked to transcriptional alterations affecting Myc and its target genes. Here, we set out to understand to what extent the role of cohesins in controlling cell cycle is dependent on Myc expression and activity. Inactivation of the cohesin complex by silencing the RAD21 subunit led to cell cycle arrest due to both transcriptional impairment of Myc target genes and alterations of replication forks, which were fewer and preferentially unidirectional. Ectopic activation of Myc in RAD21 depleted cells rescued Myc-dependent transcription and promoted S-phase entry but failed to sustain S-phase progression due to a strong replicative stress response, which was associated to a robust DNA damage response, DNA damage checkpoint activation and synthetic lethality. Thus, the cohesin complex is dispensable for Myc-dependent transcription but essential to prevent Myc-induced replicative stress. This suggests the presence of a feed-forward regulatory loop where cohesins by regulating Myc level control S-phase entry and prevent replicative stress. |
Puccio S; Grillo G; Licciulli F; Severgnini M; Liuni S; Bicciato S; De Bellis G; Ferrari F; Peano C WoPPER: Web server for Position Related data analysis of gene Expression in Prokaryotes. Journal Article In: Nucleic Acids Research, vol. 45, no W1, pp. W109-W115, 2017. @article{%a1:%Y_183,
title = {WoPPER: Web server for Position Related data analysis of gene Expression in Prokaryotes.},
author = {Puccio S and Grillo G and Licciulli F and Severgnini M and Liuni S and Bicciato S and De Bellis G and Ferrari F and Peano C},
url = {https://academic.oup.com/nar/article/45/W1/W109/3782601},
doi = {10.1093/nar/gkx329},
year = {2017},
date = {2017-07-13},
journal = {Nucleic Acids Research},
volume = {45},
number = {W1},
pages = {W109-W115},
abstract = {The structural and conformational organization of chromosomes is crucial for gene expression regulation in eukaryotes and prokaryotes as well. Up to date, gene expression data generated using either microarray or RNA-sequencing are available for many bacterial genomes. However, differential gene expression is usually investigated with methods considering each gene independently, thus not taking into account the physical localization of genes along a bacterial chromosome. Here, we present WoPPER, a web tool integrating gene expression and genomic annotations to identify differentially expressed chromosomal regions in bacteria. RNA-sequencing or microarray-based gene expression data are provided as input, along with gene annotations. The user can select genomic annotations from an internal database including 2780 bacterial strains, or provide custom genomic annotations. The analysis produces as output the lists of positionally related genes showing a coordinated trend of differential expression. Graphical representations, including a circular plot of the analyzed chromosome, allow intuitive browsing of the results. The analysis procedure is based on our previously published R-package PREDA. The release of this tool is timely and relevant for the scientific community, as WoPPER will fill an existing gap in prokaryotic gene expression data analysis and visualization tools. WoPPER is open to all users and can be reached at the following URL: https://WoPPER.ba.itb.cnr.it.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The structural and conformational organization of chromosomes is crucial for gene expression regulation in eukaryotes and prokaryotes as well. Up to date, gene expression data generated using either microarray or RNA-sequencing are available for many bacterial genomes. However, differential gene expression is usually investigated with methods considering each gene independently, thus not taking into account the physical localization of genes along a bacterial chromosome. Here, we present WoPPER, a web tool integrating gene expression and genomic annotations to identify differentially expressed chromosomal regions in bacteria. RNA-sequencing or microarray-based gene expression data are provided as input, along with gene annotations. The user can select genomic annotations from an internal database including 2780 bacterial strains, or provide custom genomic annotations. The analysis produces as output the lists of positionally related genes showing a coordinated trend of differential expression. Graphical representations, including a circular plot of the analyzed chromosome, allow intuitive browsing of the results. The analysis procedure is based on our previously published R-package PREDA. The release of this tool is timely and relevant for the scientific community, as WoPPER will fill an existing gap in prokaryotic gene expression data analysis and visualization tools. WoPPER is open to all users and can be reached at the following URL: https://WoPPER.ba.itb.cnr.it. |
Vetro A; Savasta S; Russo Raucci A; Cerqua C; Sartori G; Limongelli I; Forlino A; Maruelli S; Perucca P; Vergani D; Mazzini G; Mattevi A; Stivala LA; Salviati L; Zuffardi O MCM5: a new actor in the link between DNA replication and Meier-Gorlin syndrome. Journal Article In: European journal of human genetics, vol. 25, no 5, pp. 646-650, 2017. @article{%a1:%Y_221,
title = {MCM5: a new actor in the link between DNA replication and Meier-Gorlin syndrome.},
author = {Vetro A and Savasta S and Russo Raucci A and Cerqua C and Sartori G and Limongelli I and Forlino A and Maruelli S and Perucca P and Vergani D and Mazzini G and Mattevi A and Stivala LA and Salviati L and Zuffardi O},
url = {http://www.nature.com/ejhg/journal/vaop/ncurrent/full/ejhg20175a.html},
doi = {10.1038/ejhg.2017.5},
year = {2017},
date = {2017-05-18},
journal = {European journal of human genetics},
volume = {25},
number = {5},
pages = {646-650},
abstract = {Meier-Gorlin syndrome (MGORS) is a rare disorder characterized by primordial dwarfism, microtia, and patellar aplasia/hypoplasia. Recessive mutations in ORC1, ORC4, ORC6, CDT1, CDC6, and CDC45, encoding members of the pre-replication (pre-RC) and pre-initiation (pre-IC) complexes, and heterozygous mutations in GMNN, a regulator of cell-cycle progression and DNA replication, have already been associated with this condition. We performed whole-exome sequencing (WES) in a patient with a clinical diagnosis of MGORS and identified biallelic variants in MCM5. This gene encodes a subunit of the replicative helicase complex, which represents a component of the pre-RC. Both variants, a missense substitution within a conserved domain critical for the helicase activity, and a single base deletion causing a frameshift and a premature stop codon, were predicted to be detrimental for the MCM5 function. Although variants of MCM5 have never been reported in specific human diseases, defect of this gene in zebrafish causes a phenotype of growth restriction overlapping the one associated with orc1 depletion. Complementation experiments in yeast showed that the plasmid carrying the missense variant was unable to rescue the lethal phenotype caused by mcm5 deletion. Moreover cell-cycle progression was delayed in patient's cells, as already shown for mutations in the ORC1 gene. Altogether our findings support the role of MCM5 as a novel gene involved in MGORS, further emphasizing that this condition is caused by impaired DNA replication.European Journal of Human Genetics advance online publication, 15 February 2017; doi:10.1038/ejhg.2017.5.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Meier-Gorlin syndrome (MGORS) is a rare disorder characterized by primordial dwarfism, microtia, and patellar aplasia/hypoplasia. Recessive mutations in ORC1, ORC4, ORC6, CDT1, CDC6, and CDC45, encoding members of the pre-replication (pre-RC) and pre-initiation (pre-IC) complexes, and heterozygous mutations in GMNN, a regulator of cell-cycle progression and DNA replication, have already been associated with this condition. We performed whole-exome sequencing (WES) in a patient with a clinical diagnosis of MGORS and identified biallelic variants in MCM5. This gene encodes a subunit of the replicative helicase complex, which represents a component of the pre-RC. Both variants, a missense substitution within a conserved domain critical for the helicase activity, and a single base deletion causing a frameshift and a premature stop codon, were predicted to be detrimental for the MCM5 function. Although variants of MCM5 have never been reported in specific human diseases, defect of this gene in zebrafish causes a phenotype of growth restriction overlapping the one associated with orc1 depletion. Complementation experiments in yeast showed that the plasmid carrying the missense variant was unable to rescue the lethal phenotype caused by mcm5 deletion. Moreover cell-cycle progression was delayed in patient's cells, as already shown for mutations in the ORC1 gene. Altogether our findings support the role of MCM5 as a novel gene involved in MGORS, further emphasizing that this condition is caused by impaired DNA replication.European Journal of Human Genetics advance online publication, 15 February 2017; doi:10.1038/ejhg.2017.5. |
D'Auria F; Centurione L; Centurione MA; Angelini A; Di Pietro R Regulation of Cancer Cell Responsiveness to Ionizing Radiation Treatment by Cyclic AMP Response Element Binding Nuclear Transcription Factor. Journal Article In: Frontiers in oncology, vol. 7, no 1, pp. 76, 2017. @article{%a1:%Y_212,
title = {Regulation of Cancer Cell Responsiveness to Ionizing Radiation Treatment by Cyclic AMP Response Element Binding Nuclear Transcription Factor.},
author = {D'Auria F and Centurione L and Centurione MA and Angelini A and Di Pietro R},
url = {https://www.frontiersin.org/articles/10.3389/fonc.2017.00076/full},
doi = {10.3389/fonc.2017.00076},
year = {2017},
date = {2017-05-10},
journal = {Frontiers in oncology},
volume = {7},
number = {1},
pages = {76},
abstract = {Cyclic AMP response element binding (CREB) protein is a member of the CREB/activating transcription factor (ATF) family of transcription factors that play an important role in the cell response to different environmental stimuli leading to proliferation, differentiation, apoptosis, and survival. A number of studies highlight the involvement of CREB in the resistance to ionizing radiation (IR) therapy, demonstrating a relationship between IR-induced CREB family members' activation and cell survival. Consistent with these observations, we have recently demonstrated that CREB and ATF-1 are expressed in leukemia cell lines and that low-dose radiation treatment can trigger CREB activation, leading to survival of erythro-leukemia cells (K562). On the other hand, a number of evidences highlight a proapoptotic role of CREB following IR treatment of cancer cells. Since the development of multiple mechanisms of resistance is one key problem of most malignancies, including those of hematological origin, it is highly desirable to identify biological markers of responsiveness/unresponsiveness useful to follow-up the individual response and to adjust anticancer treatments. Taking into account all these considerations, this mini-review will be focused on the involvement of CREB/ATF family members in response to IR therapy, to deepen our knowledge of this topic, and to pave the way to translation into a therapeutic context.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Cyclic AMP response element binding (CREB) protein is a member of the CREB/activating transcription factor (ATF) family of transcription factors that play an important role in the cell response to different environmental stimuli leading to proliferation, differentiation, apoptosis, and survival. A number of studies highlight the involvement of CREB in the resistance to ionizing radiation (IR) therapy, demonstrating a relationship between IR-induced CREB family members' activation and cell survival. Consistent with these observations, we have recently demonstrated that CREB and ATF-1 are expressed in leukemia cell lines and that low-dose radiation treatment can trigger CREB activation, leading to survival of erythro-leukemia cells (K562). On the other hand, a number of evidences highlight a proapoptotic role of CREB following IR treatment of cancer cells. Since the development of multiple mechanisms of resistance is one key problem of most malignancies, including those of hematological origin, it is highly desirable to identify biological markers of responsiveness/unresponsiveness useful to follow-up the individual response and to adjust anticancer treatments. Taking into account all these considerations, this mini-review will be focused on the involvement of CREB/ATF family members in response to IR therapy, to deepen our knowledge of this topic, and to pave the way to translation into a therapeutic context. |
Azzalin A; Nato G; Parmigiani E; Garello F; Buffo A; Magrassi L Inhibitors of GLUT/SLC2A Enhance the Action of BCNU and Temozolomide against High-Grade Gliomas. Journal Article In: Neoplasia, vol. 19, no 4, pp. 364-373, 2017. @article{%a1:%Y_205,
title = {Inhibitors of GLUT/SLC2A Enhance the Action of BCNU and Temozolomide against High-Grade Gliomas.},
author = {Azzalin A and Nato G and Parmigiani E and Garello F and Buffo A and Magrassi L},
url = {http://www.neoplasia.com/article/S1476-5586(16)30347-5/fulltext},
doi = {10.1016/j.neo.2017.02.009},
year = {2017},
date = {2017-04-27},
journal = {Neoplasia},
volume = {19},
number = {4},
pages = {364-373},
abstract = {Glucose transport across glioblastoma membranes plays a crucial role in maintaining the enhanced glycolysis typical of high-grade gliomas and glioblastoma. We tested the ability of two inhibitors of the glucose transporters GLUT/SLC2A superfamily, indinavir (IDV) and ritonavir (RTV), and of one inhibitor of the Na/glucose antiporter type 2 (SGLT2/SLC5A2) superfamily, phlorizin (PHZ), in decreasing glucose consumption and cell proliferation of human and murine glioblastoma cells. We found in vitro that RTV, active on at least three different GLUT/SLC2A transporters, was more effective than IDV, a specific inhibitor of GLUT4/SLC2A4, both in decreasing glucose consumption and lactate production and in inhibiting growth of U87MG and Hu197 human glioblastoma cell lines and primary cultures of human glioblastoma. PHZ was inactive on the same cells. Similar results were obtained when cells were grown in adherence or as 3D multicellular tumor spheroids. RTV treatment but not IDV treatment induced AMP-activated protein kinase (AMPKα) phosphorylation that paralleled the decrease in glycolytic activity and cell growth. IDV, but not RTV, induced an increase in GLUT1/SLC2A1 whose activity could compensate for the inhibition of GLUT4/SLC2A4 by IDV. RTV and IDV pass poorly the blood brain barrier and are unlikely to reach sufficient liquoral concentrations in vivo to inhibit glioblastoma growth as single agents. Isobologram analysis of the association of RTV or IDV and 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) or 4-methyl-5-oxo-2,3,4,6,8-pentazabicyclo[4.3.0]nona-2,7,9-triene-9-carboxamide (TMZ) indicated synergy only with RTV on inhibition of glioblastoma cells. Finally, we tested in vivo the combination of RTV and BCNU on established GL261 tumors. This drug combination increased the overall survival and allowed a five-fold reduction in the dose of BCNU.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Glucose transport across glioblastoma membranes plays a crucial role in maintaining the enhanced glycolysis typical of high-grade gliomas and glioblastoma. We tested the ability of two inhibitors of the glucose transporters GLUT/SLC2A superfamily, indinavir (IDV) and ritonavir (RTV), and of one inhibitor of the Na/glucose antiporter type 2 (SGLT2/SLC5A2) superfamily, phlorizin (PHZ), in decreasing glucose consumption and cell proliferation of human and murine glioblastoma cells. We found in vitro that RTV, active on at least three different GLUT/SLC2A transporters, was more effective than IDV, a specific inhibitor of GLUT4/SLC2A4, both in decreasing glucose consumption and lactate production and in inhibiting growth of U87MG and Hu197 human glioblastoma cell lines and primary cultures of human glioblastoma. PHZ was inactive on the same cells. Similar results were obtained when cells were grown in adherence or as 3D multicellular tumor spheroids. RTV treatment but not IDV treatment induced AMP-activated protein kinase (AMPKα) phosphorylation that paralleled the decrease in glycolytic activity and cell growth. IDV, but not RTV, induced an increase in GLUT1/SLC2A1 whose activity could compensate for the inhibition of GLUT4/SLC2A4 by IDV. RTV and IDV pass poorly the blood brain barrier and are unlikely to reach sufficient liquoral concentrations in vivo to inhibit glioblastoma growth as single agents. Isobologram analysis of the association of RTV or IDV and 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) or 4-methyl-5-oxo-2,3,4,6,8-pentazabicyclo[4.3.0]nona-2,7,9-triene-9-carboxamide (TMZ) indicated synergy only with RTV on inhibition of glioblastoma cells. Finally, we tested in vivo the combination of RTV and BCNU on established GL261 tumors. This drug combination increased the overall survival and allowed a five-fold reduction in the dose of BCNU. |
Ultimo S; Simioni C; Martelli AM; Zauli G; Evangelisti C; Celeghini C; McCubrey JA; Marisi G; Ulivi P; Capitani S; Neri LM PI3K isoform inhibition associated with anti Bcr-Abl drugs shows in vitro increased anti-leukemic activity in Philadelphia chromosome-positive B-acute lymphoblastic leukemia cell lines. Journal Article In: Oncotarget, vol. 8, no 14, pp. 23213-23227, 2017. @article{%a1:%Y_204,
title = {PI3K isoform inhibition associated with anti Bcr-Abl drugs shows in vitro increased anti-leukemic activity in Philadelphia chromosome-positive B-acute lymphoblastic leukemia cell lines.},
author = {Ultimo S and Simioni C and Martelli AM and Zauli G and Evangelisti C and Celeghini C and McCubrey JA and Marisi G and Ulivi P and Capitani S and Neri LM},
url = {http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path[]=15542&pubmed-linkout=1},
doi = {10.18632/oncotarget.15542},
year = {2017},
date = {2017-04-04},
journal = {Oncotarget},
volume = {8},
number = {14},
pages = {23213-23227},
abstract = {B-acute lymphoblastic leukemia (B-ALL) is a malignant disorder characterized by the abnormal proliferation of B-cell progenitors. Philadelphia chromosome-positive (Ph+) B-ALL is a subtype that expresses the Bcr-Abl fusion protein which represents a negative prognostic factor. Constitutive activation of the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) network is a common feature of B-ALL, influencing cell growth and survival. In the present study, we aimed to investigate the efficacy of PI3K isoform inhibition in B-ALL cell lines harboring the Bcr-Abl fusion protein.We studied the effects of anti Bcr-Abl drugs Imatinib, Nilotinib and GZD824 associated with PI3K isoform inhibitors. We used a panel of six compounds which specifically target PI3K isoforms including the pan-PI3K inhibitor ZSTK474, p110alpha BYL719 inhibitor and the dual p110gamma/p110delta inhibitor IPI145. The effects of single drugs and of several drug combinations were analyzed to assess cytotoxicity by MTS assays, apoptosis and autophagy by flow cytometry and Western blot, as well as the phosphorylation status of the pathway.ZSTK474, BYL719 and IPI145 administered in combination with imatinib, nilotinib and GZD824 for 48 h, decreased cell viability, induced apoptosis and autophagy in a marked synergistic manner.These findings suggest that selected PI3K isoform inhibitors used in combination with anti Bcr-Abl drugs may be an attractive novel therapeutic intervention in Ph+ B-ALL.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
B-acute lymphoblastic leukemia (B-ALL) is a malignant disorder characterized by the abnormal proliferation of B-cell progenitors. Philadelphia chromosome-positive (Ph+) B-ALL is a subtype that expresses the Bcr-Abl fusion protein which represents a negative prognostic factor. Constitutive activation of the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) network is a common feature of B-ALL, influencing cell growth and survival. In the present study, we aimed to investigate the efficacy of PI3K isoform inhibition in B-ALL cell lines harboring the Bcr-Abl fusion protein.We studied the effects of anti Bcr-Abl drugs Imatinib, Nilotinib and GZD824 associated with PI3K isoform inhibitors. We used a panel of six compounds which specifically target PI3K isoforms including the pan-PI3K inhibitor ZSTK474, p110alpha BYL719 inhibitor and the dual p110gamma/p110delta inhibitor IPI145. The effects of single drugs and of several drug combinations were analyzed to assess cytotoxicity by MTS assays, apoptosis and autophagy by flow cytometry and Western blot, as well as the phosphorylation status of the pathway.ZSTK474, BYL719 and IPI145 administered in combination with imatinib, nilotinib and GZD824 for 48 h, decreased cell viability, induced apoptosis and autophagy in a marked synergistic manner.These findings suggest that selected PI3K isoform inhibitors used in combination with anti Bcr-Abl drugs may be an attractive novel therapeutic intervention in Ph+ B-ALL. |
Brogi S; Ramunno A; Savi L; Chemi G; Alfano G; Pecorelli A; Pambianchi E; Galatello P; Compagnoni G; Focher F; Biamonti G; Valacchi G; Butini S; Gemma S; Campiani G; Brindisi M First dual AK/GSK-3beta inhibitors endowed with antioxidant properties as multifunctional, potential neuroprotective agents. Journal Article In: European journal of medicinal chemistry, vol. 138, pp. 438-457, 2017. @article{%a1:%Y_208,
title = {First dual AK/GSK-3beta inhibitors endowed with antioxidant properties as multifunctional, potential neuroprotective agents.},
author = {Brogi S and Ramunno A and Savi L and Chemi G and Alfano G and Pecorelli A and Pambianchi E and Galatello P and Compagnoni G and Focher F and Biamonti G and Valacchi G and Butini S and Gemma S and Campiani G and Brindisi M},
url = {http://www.sciencedirect.com/science/article/pii/S0223523417304634?via%3Dihub},
doi = {10.1016/j.ejmech.2017.06.017},
year = {2017},
date = {2017-03-24},
journal = {European journal of medicinal chemistry},
volume = {138},
pages = {438-457},
abstract = {The manuscript deals with the design, synthesis and biological evaluation of novel benzoxazinone-based and indole-based compounds as multifunctional neuroprotective agents. These compounds inhibit human adenosine kinase (hAK) and human glycogen synthase kinase 3 beta (hGSK-3β) enzymes. Computational analysis based on a molecular docking approach underlined the potential structural requirements for simultaneously targeting both proteins' allosteric sites. In silico hints drove the synthesis of appropriately decorated benzoxazinones and indoles (5a-s, and 6a-c) and biochemical analysis revealed their behavior as allosteric inhibitors of hGSK-3beta. For both our hit 4 and the best compounds of the series (5c,l and 6b) the potential antioxidant profile was assessed in human neuroblastoma cell lines (IMR 32, undifferentiated and neuronal differentiated), by evaluating the protective effect of selected compounds against H2O2 cytotoxicity and reactive oxygen species (ROS) production. Results showed a strong efficacy of the tested compounds, even at the lower doses, in counteracting the induced oxidative stress (50 μM of H2O2) and in preventing ROS formation. In addition, the tested compounds did not show any cytotoxic effect determined by the LDH release, at the concentration range analyzed (from 0.1 to 50 μM). This study allowed the identification of compound 5l, as the first dual hAK/hGSK-3β inhibitor reported to date. Compound 5l, which behaves as an effective antioxidant, holds promise for the development of new series of potential therapeutic agents for the treatment of neurodegenerative diseases characterized by an innovative pharmacological profile.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The manuscript deals with the design, synthesis and biological evaluation of novel benzoxazinone-based and indole-based compounds as multifunctional neuroprotective agents. These compounds inhibit human adenosine kinase (hAK) and human glycogen synthase kinase 3 beta (hGSK-3β) enzymes. Computational analysis based on a molecular docking approach underlined the potential structural requirements for simultaneously targeting both proteins' allosteric sites. In silico hints drove the synthesis of appropriately decorated benzoxazinones and indoles (5a-s, and 6a-c) and biochemical analysis revealed their behavior as allosteric inhibitors of hGSK-3beta. For both our hit 4 and the best compounds of the series (5c,l and 6b) the potential antioxidant profile was assessed in human neuroblastoma cell lines (IMR 32, undifferentiated and neuronal differentiated), by evaluating the protective effect of selected compounds against H2O2 cytotoxicity and reactive oxygen species (ROS) production. Results showed a strong efficacy of the tested compounds, even at the lower doses, in counteracting the induced oxidative stress (50 μM of H2O2) and in preventing ROS formation. In addition, the tested compounds did not show any cytotoxic effect determined by the LDH release, at the concentration range analyzed (from 0.1 to 50 μM). This study allowed the identification of compound 5l, as the first dual hAK/hGSK-3β inhibitor reported to date. Compound 5l, which behaves as an effective antioxidant, holds promise for the development of new series of potential therapeutic agents for the treatment of neurodegenerative diseases characterized by an innovative pharmacological profile. |
Cenni V Letter to the editor: Comments on Wette et al. (2017): "Characterization of muscle ankyrin repeat proteins in human skeletal muscle". Journal Article In: American journal of physiology. Cell physiology, vol. 313, no 4, pp. C469-C470, 2017. @article{%a1:%Y_223,
title = {Letter to the editor: Comments on Wette et al. (2017): "Characterization of muscle ankyrin repeat proteins in human skeletal muscle".},
author = {Cenni V},
url = {http://ajpcell.physiology.org/content/313/4/C469.long},
doi = {10.1152/ajpcell.00151.2017},
year = {2017},
date = {2017-03-23},
journal = {American journal of physiology. Cell physiology},
volume = {313},
number = {4},
pages = {C469-C470},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
|
Pregnolato M; Damiani G; Pereira A Jr Patterns of calcium signaling: A link between chronic emotions and cancer. Journal Article In: Journal of integrative neuroscience, vol. 16, no s1, pp. s43-s-63, 2017. @article{%a1:%Y_239,
title = {Patterns of calcium signaling: A link between chronic emotions and cancer.},
author = {Pregnolato M and Damiani G and Pereira A Jr},
url = {https://content.iospress.com/articles/journal-of-integrative-neuroscience/jin066},
doi = {10.3233/JIN-170066},
year = {2017},
date = {2017-03-23},
journal = {Journal of integrative neuroscience},
volume = {16},
number = {s1},
pages = {s43-s-63},
abstract = {Intra and inter-cellular calcium signaling is present in all types of cells and body tissues. In the human brain, calcium currents and waves are related to mental activities, including emotions. We present a theoretical interpretation of these phenomena suggesting their involvement in chronic emotional patterns and in the pathology of cancer. Recent developments on biophysics, translational biology and psychoneuroendocrinoimmunology (PNEI) can support explanatory hypotheses about the link between emotional stresses and the origin and development of different types of tumor cells. Chronic stresses may cause perturbations of rhythms of the PNEI system, excessive activation of HPA axis and abnormal activation of calcium signals in somatic tissues, with deleterious effects on different parts of the body. The increasing of calcium signaling inside cells may lead to a deregulation of different pathways and epigenetic systems that promote the production of genomic mutations in a second phase. In particular, the hyperactivation of the transcription nuclear factor kappaB (NF-κB), if is not counterbalanced by the following activation of the nuclear factor (erythroid-derived 2)-like 2 (NFE2L2 or Nrf2), increases the production of oxidative catabolites, as the advanced glycation end products (AGE), which play a key role in the progression of different types of cancer and other degenerative diseases. Cortisol binding to glucocorticoid receptor (GR) reduces the activity of both NF-κB and Nrf2 inside the cells but inhibits the cellular immunity and the anabolic processes of tissue regeneration. The tissue atrophy and the defective anti-ageing mechanisms promotes the tumoral cells growth and their escape from the immune-surveillance.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Intra and inter-cellular calcium signaling is present in all types of cells and body tissues. In the human brain, calcium currents and waves are related to mental activities, including emotions. We present a theoretical interpretation of these phenomena suggesting their involvement in chronic emotional patterns and in the pathology of cancer. Recent developments on biophysics, translational biology and psychoneuroendocrinoimmunology (PNEI) can support explanatory hypotheses about the link between emotional stresses and the origin and development of different types of tumor cells. Chronic stresses may cause perturbations of rhythms of the PNEI system, excessive activation of HPA axis and abnormal activation of calcium signals in somatic tissues, with deleterious effects on different parts of the body. The increasing of calcium signaling inside cells may lead to a deregulation of different pathways and epigenetic systems that promote the production of genomic mutations in a second phase. In particular, the hyperactivation of the transcription nuclear factor kappaB (NF-κB), if is not counterbalanced by the following activation of the nuclear factor (erythroid-derived 2)-like 2 (NFE2L2 or Nrf2), increases the production of oxidative catabolites, as the advanced glycation end products (AGE), which play a key role in the progression of different types of cancer and other degenerative diseases. Cortisol binding to glucocorticoid receptor (GR) reduces the activity of both NF-κB and Nrf2 inside the cells but inhibits the cellular immunity and the anabolic processes of tissue regeneration. The tissue atrophy and the defective anti-ageing mechanisms promotes the tumoral cells growth and their escape from the immune-surveillance. |
Mentegari E; Crespan E; Bavagnoli L; Kissova M; Bertoletti F; Sabbioneda S; Imhof R; Sturla SJ; Nilforoushan A; Hubscher U; van Loon B; Maga G Ribonucleotide incorporation by human DNA polymerase eta impacts translesion synthesis and RNase H2 activity. Journal Article In: Nucleic Acids Research, vol. 45, no 5, pp. 2600-2614, 2017. @article{%a1:%Y_190,
title = {Ribonucleotide incorporation by human DNA polymerase eta impacts translesion synthesis and RNase H2 activity.},
author = {Mentegari E and Crespan E and Bavagnoli L and Kissova M and Bertoletti F and Sabbioneda S and Imhof R and Sturla SJ and Nilforoushan A and Hubscher U and van Loon B and Maga G},
url = {https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/gkw1275},
doi = {doi.org/10.1093/nar/gkw1275},
year = {2017},
date = {2017-03-16},
journal = {Nucleic Acids Research},
volume = {45},
number = {5},
pages = {2600-2614},
abstract = {Ribonucleotides (rNs) incorporated in the genome by DNA polymerases (Pols) are removed by RNase H2. Cytidine and guanosine preferentially accumulate over the other rNs. Here we show that human Pol η can incorporate cytidine monophosphate (rCMP) opposite guanine, 8-oxo-7,8-dihydroguanine, 8-methyl-2'-deoxyguanosine and a cisplatin intrastrand guanine crosslink (cis-PtGG), while it cannot bypass a 3-methylcytidine or an abasic site with rNs as substrates. Pol eta is also capable of synthesizing polyribonucleotide chains, and its activity is enhanced by its auxiliary factor DNA Pol delta interacting protein 2 (PolDIP2). Human RNase H2 removes cytidine and guanosine less efficiently than the other rNs and incorporation of rCMP opposite DNA lesions further reduces the efficiency of RNase H2. Experiments with XP-V cell extracts indicate Pol eta as the major basis of rCMP incorporation opposite cis-PtGG. These results suggest that translesion synthesis by Pol eta can contribute to the accumulation of rCMP in the genome, particularly opposite modified guanines.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Ribonucleotides (rNs) incorporated in the genome by DNA polymerases (Pols) are removed by RNase H2. Cytidine and guanosine preferentially accumulate over the other rNs. Here we show that human Pol η can incorporate cytidine monophosphate (rCMP) opposite guanine, 8-oxo-7,8-dihydroguanine, 8-methyl-2'-deoxyguanosine and a cisplatin intrastrand guanine crosslink (cis-PtGG), while it cannot bypass a 3-methylcytidine or an abasic site with rNs as substrates. Pol eta is also capable of synthesizing polyribonucleotide chains, and its activity is enhanced by its auxiliary factor DNA Pol delta interacting protein 2 (PolDIP2). Human RNase H2 removes cytidine and guanosine less efficiently than the other rNs and incorporation of rCMP opposite DNA lesions further reduces the efficiency of RNase H2. Experiments with XP-V cell extracts indicate Pol eta as the major basis of rCMP incorporation opposite cis-PtGG. These results suggest that translesion synthesis by Pol eta can contribute to the accumulation of rCMP in the genome, particularly opposite modified guanines. |
Zanin E; Riva M; Bambini V; Cappa SF; Magrassi L; Moro A The contribution of surgical brain mapping to the understanding of the anatomo-functional basis of syntax: A critical review. Journal Article In: Neurological Sciences, vol. 38, no 9, pp. 1579-1589, 2017. @article{%a1:%Y_228,
title = {The contribution of surgical brain mapping to the understanding of the anatomo-functional basis of syntax: A critical review.},
author = {Zanin E and Riva M and Bambini V and Cappa SF and Magrassi L and Moro A},
url = {https://link.springer.com/article/10.1007%2Fs10072-017-3016-4},
doi = {10.1007/s10072-017-3016-4},
year = {2017},
date = {2017-03-15},
journal = {Neurological Sciences},
volume = {38},
number = {9},
pages = {1579-1589},
abstract = {A wide range of studies on language assessment during awake brain surgery is nowadays available. Yet, a consensus on a standardized protocol for intraoperative language mapping is still lacking. More specifically, very limited information is offered about intraoperative assessment of a crucial component of language such as syntax. This review aims at critically analyzing the intraoperative studies investigating the cerebral basis of syntactic processing. A comprehensive query was performed on the literature, returning a total of 18 studies. These papers were analyzed according to two complementary criteria, based on the distinction between morphosyntax and syntax. The first criterion focused on the tasks and stimuli employed intraoperatively. Studies were divided into three different groups: group 1 included those studies that overtly aimed at investigating morphosyntactic processes; group 2 included studies that did not explicitly focus on syntax, yet employed stimuli requiring morphosyntactic processing; and group 3 included studies reporting some generic form of syntactic deficit, although not further investigated. The second criterion focused on the syntactic structures of the sentences assessed intraoperatively, analyzing the canonicity of sentence structure (i.e., canonical versus non-canonical word order). The global picture emerging from our analysis indicates that what was investigated in the intraoperative literature is morphosyntactic processing, rather than pure syntax. The study of the neurobiology of syntax during awake surgery seems thus to be still at an early stage, in need of systematic, linguistically grounded investigations.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
A wide range of studies on language assessment during awake brain surgery is nowadays available. Yet, a consensus on a standardized protocol for intraoperative language mapping is still lacking. More specifically, very limited information is offered about intraoperative assessment of a crucial component of language such as syntax. This review aims at critically analyzing the intraoperative studies investigating the cerebral basis of syntactic processing. A comprehensive query was performed on the literature, returning a total of 18 studies. These papers were analyzed according to two complementary criteria, based on the distinction between morphosyntax and syntax. The first criterion focused on the tasks and stimuli employed intraoperatively. Studies were divided into three different groups: group 1 included those studies that overtly aimed at investigating morphosyntactic processes; group 2 included studies that did not explicitly focus on syntax, yet employed stimuli requiring morphosyntactic processing; and group 3 included studies reporting some generic form of syntactic deficit, although not further investigated. The second criterion focused on the syntactic structures of the sentences assessed intraoperatively, analyzing the canonicity of sentence structure (i.e., canonical versus non-canonical word order). The global picture emerging from our analysis indicates that what was investigated in the intraoperative literature is morphosyntactic processing, rather than pure syntax. The study of the neurobiology of syntax during awake surgery seems thus to be still at an early stage, in need of systematic, linguistically grounded investigations. |
Robuffo I; Toniato E; Tettamanti L; Mastrangelo F; Ronconi G; Frydas I; Caraffa A; Kritas SK; Conti P Mast cell in innate immunity mediated by proinflammatory and antiinflammatory IL-1 family members. Journal Article In: Journal of biological regulators and homeostatic agents, vol. 31, no 4, pp. 837-842, 2017. @article{%a1:%Y_236,
title = {Mast cell in innate immunity mediated by proinflammatory and antiinflammatory IL-1 family members.},
author = {Robuffo I and Toniato E and Tettamanti L and Mastrangelo F and Ronconi G and Frydas I and Caraffa A and Kritas SK and Conti P},
url = {https://www.biolifesas.org/biolife/category/journals/journal-of-biological-regulators-and-homeostatic-agents/},
year = {2017},
date = {2017-03-15},
journal = {Journal of biological regulators and homeostatic agents},
volume = {31},
number = {4},
pages = {837-842},
abstract = {Innate immunity consists of physical and chemical barriers which provide the early defense against infections. Innate immunity orchestrates the defense of the host with cellular and biochemical proteins. Mast cells (MCs) are involved in innate and adaptive immunity and are the first line of defense which generates multiple inflammatory cytokines/chemokines in response to numerous antigens. MC-activated antigen receptor Fc-RI provokes a number of important biochemical pathways with secretion of numerous vasoactive, chemoattractant and inflammatory compounds which participate in allergic and inflammatory diseases. MCs can also be activated by Th1 cytokines and generate pre-formed and de novo inflammatory mediators, including TNF. IL-37 is an anti-inflammatory cytokine which binds IL-18R-alpha chain and reduces the production of inflammatory IL-1 family members. IL-37 down-regulates innate immunity by inhibiting macrophage response and its accumulation and reduces the cytokines that mediate inflammatory diseases. Here, we discuss the relationship between MCs, innate immunity, and pro-inflammatory and anti-inflammatory cytokines.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Innate immunity consists of physical and chemical barriers which provide the early defense against infections. Innate immunity orchestrates the defense of the host with cellular and biochemical proteins. Mast cells (MCs) are involved in innate and adaptive immunity and are the first line of defense which generates multiple inflammatory cytokines/chemokines in response to numerous antigens. MC-activated antigen receptor Fc-RI provokes a number of important biochemical pathways with secretion of numerous vasoactive, chemoattractant and inflammatory compounds which participate in allergic and inflammatory diseases. MCs can also be activated by Th1 cytokines and generate pre-formed and de novo inflammatory mediators, including TNF. IL-37 is an anti-inflammatory cytokine which binds IL-18R-alpha chain and reduces the production of inflammatory IL-1 family members. IL-37 down-regulates innate immunity by inhibiting macrophage response and its accumulation and reduces the cytokines that mediate inflammatory diseases. Here, we discuss the relationship between MCs, innate immunity, and pro-inflammatory and anti-inflammatory cytokines. |
Meroni A; Mentegari E; Crespan E; Muzi-Falconi M; Lazzaro F; Podestà A The Incorporation of Ribonucleotides Induces Structural and Conformational Changes in DNA. Journal Article In: Biophysical journal, vol. 113, no 7, pp. 1373-1382, 2017. @article{%a1:%Y_222,
title = {The Incorporation of Ribonucleotides Induces Structural and Conformational Changes in DNA.},
author = {Meroni A and Mentegari E and Crespan E and Muzi-Falconi M and Lazzaro F and Podestà A},
url = {https://www.ncbi.nlm.nih.gov/pubmed/28978432},
doi = {10.1016/j.bpj.2017.07.013},
year = {2017},
date = {2017-03-08},
journal = {Biophysical journal},
volume = {113},
number = {7},
pages = {1373-1382},
abstract = {Ribonucleotide incorporation is the most common error occurring during DNA replication. Cells have hence developed mechanisms to remove ribonucleotides from the genome and restore its integrity. Indeed, the persistence of ribonucleotides into DNA leads to severe consequences, such as genome instability and replication stress. Thus, it becomes important to understand the effects of ribonucleotides incorporation, starting from their impact on DNA structure and conformation. Here we present a systematic study of the effects of ribonucleotide incorporation into DNA molecules. We have developed, to our knowledge, a new method to efficiently synthesize long DNA molecules (hundreds of basepairs) containing ribonucleotides, which is based on a modified protocol for the polymerase chain reaction. By means of atomic force microscopy, we could therefore investigate the changes, upon ribonucleotide incorporation, of the structural and conformational properties of numerous DNA populations at the single-molecule level. Specifically, we characterized the scaling of the contour length with the number of basepairs and the scaling of the end-to-end distance with the curvilinear distance, the bending angle distribution, and the persistence length. Our results revealed that ribonucleotides affect DNA structure and conformation on scales that go well beyond the typical dimension of the single ribonucleotide. In particular, the presence of ribonucleotides induces a systematic shortening of the molecules, together with a decrease of the persistence length. Such structural changes are also likely to occur in vivo, where they could directly affect the downstream DNA transactions, as well as interfere with protein binding and recognition.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Ribonucleotide incorporation is the most common error occurring during DNA replication. Cells have hence developed mechanisms to remove ribonucleotides from the genome and restore its integrity. Indeed, the persistence of ribonucleotides into DNA leads to severe consequences, such as genome instability and replication stress. Thus, it becomes important to understand the effects of ribonucleotides incorporation, starting from their impact on DNA structure and conformation. Here we present a systematic study of the effects of ribonucleotide incorporation into DNA molecules. We have developed, to our knowledge, a new method to efficiently synthesize long DNA molecules (hundreds of basepairs) containing ribonucleotides, which is based on a modified protocol for the polymerase chain reaction. By means of atomic force microscopy, we could therefore investigate the changes, upon ribonucleotide incorporation, of the structural and conformational properties of numerous DNA populations at the single-molecule level. Specifically, we characterized the scaling of the contour length with the number of basepairs and the scaling of the end-to-end distance with the curvilinear distance, the bending angle distribution, and the persistence length. Our results revealed that ribonucleotides affect DNA structure and conformation on scales that go well beyond the typical dimension of the single ribonucleotide. In particular, the presence of ribonucleotides induces a systematic shortening of the molecules, together with a decrease of the persistence length. Such structural changes are also likely to occur in vivo, where they could directly affect the downstream DNA transactions, as well as interfere with protein binding and recognition. |
Bavelloni A; Focaccia E; Piazzi M; Errani C; Blalock WL; Faenza I Cell Cycle Arrest and Apoptosis Induced by Kinamycin F in Human Osteosarcoma Cells. Journal Article In: Anticancer Research, vol. 37, no 8, pp. 4103-4109, 2017. @article{%a1:%Y_231,
title = {Cell Cycle Arrest and Apoptosis Induced by Kinamycin F in Human Osteosarcoma Cells.},
author = {Bavelloni A and Focaccia E and Piazzi M and Errani C and Blalock WL and Faenza I},
url = {http://ar.iiarjournals.org/content/37/8/4103.long},
year = {2017},
date = {2017-03-07},
journal = {Anticancer Research},
volume = {37},
number = {8},
pages = {4103-4109},
abstract = {Background/Aim: Kinamycin F is a bacterial metabolite which contains an unusual and potentially reactive diazo group that is known for its ability to inhibit cell growth. In this study, the potential anti-tumor activity of kinamycin F was investigated in three human osteosarcoma cell lines, MG-63, U-2 OS and HOS as an antitumor agent with a potentially novel target. Materials and Methods: Proliferation and cell viability were measured in three human osteosarcoma cell lines by commercially available kits. We also evaluated the effects of the drug on cell cycle progression using the Muse™ Cell Analyzer. Caspase-3 activity was determined by a fluorometric EnzChek assay kit. Finally, following treatment with kinamycin F the protein levels of cyclin D3, cyclin A and cdK-2 were examined. Results: Kinamycin F induced a concentration-dependent cell death in all the three cell lines. Flow cytometry revealed that kinamycin F treatment at 1 μM concentration significantly increased the cell population in the G2/M-phase (60-65%). Kinamycin F activated caspase 3 in all the three cell lines, clearly demonstrating that the growth inhibitory effect of kinamycin F can be attributed to apoptosis induction. Finally, kinamycin F suppressed osteosarcoma cell proliferation affecting cyclin A and D3 expression. Conclusion: Understanding the mechanism by which kinamycin F exerts its ability to inhibit cell growth may be a step forward in the development of new therapeutic strategies for the treatment of OS.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Background/Aim: Kinamycin F is a bacterial metabolite which contains an unusual and potentially reactive diazo group that is known for its ability to inhibit cell growth. In this study, the potential anti-tumor activity of kinamycin F was investigated in three human osteosarcoma cell lines, MG-63, U-2 OS and HOS as an antitumor agent with a potentially novel target. Materials and Methods: Proliferation and cell viability were measured in three human osteosarcoma cell lines by commercially available kits. We also evaluated the effects of the drug on cell cycle progression using the Muse™ Cell Analyzer. Caspase-3 activity was determined by a fluorometric EnzChek assay kit. Finally, following treatment with kinamycin F the protein levels of cyclin D3, cyclin A and cdK-2 were examined. Results: Kinamycin F induced a concentration-dependent cell death in all the three cell lines. Flow cytometry revealed that kinamycin F treatment at 1 μM concentration significantly increased the cell population in the G2/M-phase (60-65%). Kinamycin F activated caspase 3 in all the three cell lines, clearly demonstrating that the growth inhibitory effect of kinamycin F can be attributed to apoptosis induction. Finally, kinamycin F suppressed osteosarcoma cell proliferation affecting cyclin A and D3 expression. Conclusion: Understanding the mechanism by which kinamycin F exerts its ability to inhibit cell growth may be a step forward in the development of new therapeutic strategies for the treatment of OS. |
Angori S; Capanni C; Faulkner G; Bean C; Boriani G; Lattanzi G; Cenni V Emery-Dreifuss Muscular Dystrophy-Associated Mutant Forms of Lamin A Recruit the Stress Responsive Protein Ankrd2 into the Nucleus, Affecting the Cellular Response to Oxidative Stress. Journal Article In: Cellular Physiology and Biochemistry, vol. 42, pp. 169-184, 2017. @article{%a1:%Y_202,
title = {Emery-Dreifuss Muscular Dystrophy-Associated Mutant Forms of Lamin A Recruit the Stress Responsive Protein Ankrd2 into the Nucleus, Affecting the Cellular Response to Oxidative Stress.},
author = {Angori S and Capanni C and Faulkner G and Bean C and Boriani G and Lattanzi G and Cenni V},
url = {https://www.karger.com/Article/FullText/477309},
doi = {10.1159/000477309},
year = {2017},
date = {2017-03-07},
urldate = {2017-03-07},
journal = {Cellular Physiology and Biochemistry},
volume = {42},
pages = {169-184},
abstract = {Ankrd2 is a stress responsive protein mainly expressed in muscle cells. Upon the application of oxidative stress, Ankrd2 translocates into the nucleus where it regulates the activity of genes involved in cellular response to stress. Emery-Dreifuss Muscular Dystrophy 2 (EDMD2) is a muscular disorder caused by mutations of the gene encoding lamin A, LMNA. As well as many phenotypic abnormalities, EDMD2 muscle cells also feature a permanent basal stress state, the underlying molecular mechanisms of which are currently unclear. METHODS: Experiments were performed in EDMD2-lamin A overexpressing cell lines and EDMD2-affected human myotubes. Oxidative stress was produced by H2O2 treatment. Co-immunoprecipitation, cellular subfractionation and immunofluorescence analysis were used to validate the relation between Ankrd2 and forms of lamin A; cellular sensibility to stress was monitored by the analysis of Reactive Oxygen Species (ROS) release and cell viability. RESULTS: Our data demonstrate that oxidative stress induces the formation of a complex between Ankrd2 and lamin A. However, EDMD2-lamin A mutants were able to bind and mislocalize Ankrd2 in the nucleus even under basal conditions. Nonetheless, cells co-expressing Ankrd2 and EDMD2-lamin A mutants were more sensitive to oxidative stress than the Ankrd2-wild type lamin A counterpart. CONCLUSIONS: For the first time, we present evidence that in muscle fibers from patients affected by EDMD2, Ankrd2 has an unusual nuclear localization. By introducing a plausible mechanism ruling this accumulation, our data hint at a novel function of Ankrd2 in the pathogenesis of EDMD2-affected cells.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Ankrd2 is a stress responsive protein mainly expressed in muscle cells. Upon the application of oxidative stress, Ankrd2 translocates into the nucleus where it regulates the activity of genes involved in cellular response to stress. Emery-Dreifuss Muscular Dystrophy 2 (EDMD2) is a muscular disorder caused by mutations of the gene encoding lamin A, LMNA. As well as many phenotypic abnormalities, EDMD2 muscle cells also feature a permanent basal stress state, the underlying molecular mechanisms of which are currently unclear. METHODS: Experiments were performed in EDMD2-lamin A overexpressing cell lines and EDMD2-affected human myotubes. Oxidative stress was produced by H2O2 treatment. Co-immunoprecipitation, cellular subfractionation and immunofluorescence analysis were used to validate the relation between Ankrd2 and forms of lamin A; cellular sensibility to stress was monitored by the analysis of Reactive Oxygen Species (ROS) release and cell viability. RESULTS: Our data demonstrate that oxidative stress induces the formation of a complex between Ankrd2 and lamin A. However, EDMD2-lamin A mutants were able to bind and mislocalize Ankrd2 in the nucleus even under basal conditions. Nonetheless, cells co-expressing Ankrd2 and EDMD2-lamin A mutants were more sensitive to oxidative stress than the Ankrd2-wild type lamin A counterpart. CONCLUSIONS: For the first time, we present evidence that in muscle fibers from patients affected by EDMD2, Ankrd2 has an unusual nuclear localization. By introducing a plausible mechanism ruling this accumulation, our data hint at a novel function of Ankrd2 in the pathogenesis of EDMD2-affected cells. |
Venkata Narayanan I; Paulsen MT; Bedi K; Berg N; Ljungman EA; Francia S; Veloso A; Magnuson B; d'Adda di Fagagna F; Wilson TE; Ljungman M Transcriptional and post-transcriptional regulation of the ionizing radiation response by ATM and p53. Journal Article In: Scientific reports, vol. 7, no 1, pp. 43598, 2017. @article{%a1:%Y_216,
title = {Transcriptional and post-transcriptional regulation of the ionizing radiation response by ATM and p53.},
author = {Venkata Narayanan I and Paulsen MT and Bedi K and Berg N and Ljungman EA and Francia S and Veloso A and Magnuson B and {d'Adda di Fagagna F} and Wilson TE and Ljungman M},
url = {www.nature.com/articles/srep43598},
doi = {10.1038/srep43598},
year = {2017},
date = {2017-03-01},
journal = {Scientific reports},
volume = {7},
number = {1},
pages = {43598},
abstract = {In response to ionizing radiation (IR), cells activate a DNA damage response (DDR) pathway to re-program gene expression. Previous studies using total cellular RNA analyses have shown that the stress kinase ATM and the transcription factor p53 are integral components required for induction of IR-induced gene expression. These studies did not distinguish between changes in RNA synthesis and RNA turnover and did not address the role of enhancer elements in DDR-mediated transcriptional regulation. To determine the contribution of synthesis and degradation of RNA and monitor the activity of enhancer elements following exposure to IR, we used the recently developed Bru-seq, BruChase-seq and BruUV-seq techniques. Our results show that ATM and p53 regulate both RNA synthesis and stability as well as enhancer element activity following exposure to IR. Importantly, many genes in the p53-signaling pathway were coordinately up-regulated by both increased synthesis and RNA stability while down-regulated genes were suppressed either by reduced synthesis or stability. Our study is the first of its kind that independently assessed the effects of ionizing radiation on transcription and post-transcriptional regulation in normal human cells.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
In response to ionizing radiation (IR), cells activate a DNA damage response (DDR) pathway to re-program gene expression. Previous studies using total cellular RNA analyses have shown that the stress kinase ATM and the transcription factor p53 are integral components required for induction of IR-induced gene expression. These studies did not distinguish between changes in RNA synthesis and RNA turnover and did not address the role of enhancer elements in DDR-mediated transcriptional regulation. To determine the contribution of synthesis and degradation of RNA and monitor the activity of enhancer elements following exposure to IR, we used the recently developed Bru-seq, BruChase-seq and BruUV-seq techniques. Our results show that ATM and p53 regulate both RNA synthesis and stability as well as enhancer element activity following exposure to IR. Importantly, many genes in the p53-signaling pathway were coordinately up-regulated by both increased synthesis and RNA stability while down-regulated genes were suppressed either by reduced synthesis or stability. Our study is the first of its kind that independently assessed the effects of ionizing radiation on transcription and post-transcriptional regulation in normal human cells. |
Michelini F; Pitchiaya S; Vitelli V; Sharma S; Gioia U; Pessina F; Cabrini M; Wang Y; Capozzo I; Iannelli F; Matti V; Francia S; Shivashankar GV; Walter NG; d'Adda di Fagagna F Damage-induced lncRNAs control the DNA damage response through interaction with DDRNAs at individual double-strand breaks. Journal Article In: Nature cell biology, vol. 19, no 2, pp. 1400-1410, 2017. @article{%a1:%Y_322,
title = {Damage-induced lncRNAs control the DNA damage response through interaction with DDRNAs at individual double-strand breaks.},
author = {Michelini F and Pitchiaya S and Vitelli V and Sharma S and Gioia U and Pessina F and Cabrini M and Wang Y and Capozzo I and Iannelli F and Matti V and Francia S and Shivashankar GV and Walter NG and {d'Adda di Fagagna F}},
url = {https://www.nature.com/articles/ncb3643},
doi = {10.1038/ncb3643},
year = {2017},
date = {2017-02-28},
journal = {Nature cell biology},
volume = {19},
number = {2},
pages = {1400-1410},
abstract = {The DNA damage response (DDR) preserves genomic integrity. Small non-coding RNAs termed DDRNAs are generated at DNA double-strand breaks (DSBs) and are critical for DDR activation. Here we show that active DDRNAs specifically localize to their damaged homologous genomic sites in a transcription-dependent manner. Following DNA damage, RNA polymerase II (RNAPII) binds to the MRE11-RAD50-NBS1 complex, is recruited to DSBs and synthesizes damage-induced long non-coding RNAs (dilncRNAs) from and towards DNA ends. DilncRNAs act both as DDRNA precursors and by recruiting DDRNAs through RNA-RNA pairing. Together, dilncRNAs and DDRNAs fuel DDR focus formation and associate with 53BP1. Accordingly, inhibition of RNAPII prevents DDRNA recruitment, DDR activation and DNA repair. Antisense oligonucleotides matching dilncRNAs and DDRNAs impair site-specific DDR focus formation and DNA repair. We propose that DDR signalling sites, in addition to sharing a common pool of proteins, individually host a unique set of site-specific RNAs necessary for DDR activation.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The DNA damage response (DDR) preserves genomic integrity. Small non-coding RNAs termed DDRNAs are generated at DNA double-strand breaks (DSBs) and are critical for DDR activation. Here we show that active DDRNAs specifically localize to their damaged homologous genomic sites in a transcription-dependent manner. Following DNA damage, RNA polymerase II (RNAPII) binds to the MRE11-RAD50-NBS1 complex, is recruited to DSBs and synthesizes damage-induced long non-coding RNAs (dilncRNAs) from and towards DNA ends. DilncRNAs act both as DDRNA precursors and by recruiting DDRNAs through RNA-RNA pairing. Together, dilncRNAs and DDRNAs fuel DDR focus formation and associate with 53BP1. Accordingly, inhibition of RNAPII prevents DDRNA recruitment, DDR activation and DNA repair. Antisense oligonucleotides matching dilncRNAs and DDRNAs impair site-specific DDR focus formation and DNA repair. We propose that DDR signalling sites, in addition to sharing a common pool of proteins, individually host a unique set of site-specific RNAs necessary for DDR activation. |
Mazzini G; Danova M Fluorochromes for DNA Staining and Quantitation. Journal Article In: Methods in molecular biology - Histochemistry of Single Molecules Methods and Protocols, vol. 1560, pp. 239-259, 2017. @article{%a1:%Y_188,
title = {Fluorochromes for DNA Staining and Quantitation.},
author = {Mazzini G and Danova M},
url = {https://link.springer.com/protocol/10.1007%2F978-1-4939-6788-9_18},
doi = {10.1007/978-1-4939-6788-9_18},
year = {2017},
date = {2017-02-28},
journal = {Methods in molecular biology - Histochemistry of Single Molecules Methods and Protocols},
volume = {1560},
pages = {239-259},
abstract = {In these last few decades the great explosion of the molecular approaches has casted a little shadow on the DNA quantitative analysis. Nevertheless DNA cytochemistry represented a long piece of history in cell biology since the advent of the Feulgen reaction. This discovery was really the milestone of the emerging quantitative cytochemistry, and scientists from all over the world produced a very large literature on this subject. This first era of quantitation (histochemistry followed by cytochemistry) started by means of absorption measurements (histophotometry and cytophotometry). The successive introduction of fluorescence microscopy gave a great boost to quantitation, making easier and faster the determination of cell components by means of cytofluorometry. The development of flow cytometry further contributed to the importance of quantitative cytochemistry. At its beginning, the mission of flow cytometry was still DNA quantitation. For a decade the Feulgen reaction had been the reference methodology for both conventional and flow cytofluorometry; the advent of Shiff-type reagents contributed to expand the variety of possible fluorochromes excitable in the entire visible spectrum as well as in the ultraviolet region. The fluorescence scenario was progressively enriched by new probes among which are the intercalating dyes which made DNA quantitation simple and fast, thus spreading it worldwide. The final explosion of cytofluorometry was made possible by the availability of a large variety of probes directly binding DNA structure. In addition, immunofluorescence allowed to correlate the cell cycle-related DNA content to other cell markers. In the clinical application of flow cytometry, this promoted the introduction of multiparametric analyses aimed at describing the cytokinetic characteristics of a given cell subpopulation defined by a specific immunophenotype setting.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
In these last few decades the great explosion of the molecular approaches has casted a little shadow on the DNA quantitative analysis. Nevertheless DNA cytochemistry represented a long piece of history in cell biology since the advent of the Feulgen reaction. This discovery was really the milestone of the emerging quantitative cytochemistry, and scientists from all over the world produced a very large literature on this subject. This first era of quantitation (histochemistry followed by cytochemistry) started by means of absorption measurements (histophotometry and cytophotometry). The successive introduction of fluorescence microscopy gave a great boost to quantitation, making easier and faster the determination of cell components by means of cytofluorometry. The development of flow cytometry further contributed to the importance of quantitative cytochemistry. At its beginning, the mission of flow cytometry was still DNA quantitation. For a decade the Feulgen reaction had been the reference methodology for both conventional and flow cytofluorometry; the advent of Shiff-type reagents contributed to expand the variety of possible fluorochromes excitable in the entire visible spectrum as well as in the ultraviolet region. The fluorescence scenario was progressively enriched by new probes among which are the intercalating dyes which made DNA quantitation simple and fast, thus spreading it worldwide. The final explosion of cytofluorometry was made possible by the availability of a large variety of probes directly binding DNA structure. In addition, immunofluorescence allowed to correlate the cell cycle-related DNA content to other cell markers. In the clinical application of flow cytometry, this promoted the introduction of multiparametric analyses aimed at describing the cytokinetic characteristics of a given cell subpopulation defined by a specific immunophenotype setting. |
Croce AC; Ferrigno A; Di Pasqua LG; Berardo C; Mannucci B; Bottiroli G; Vairetti M Fluorescing fatty acids in rat fatty liver models. Journal Article In: Journal of biophotonics, vol. 10, no 6-7, pp. 905-910, 2017. @article{%a1:%Y_220,
title = {Fluorescing fatty acids in rat fatty liver models.},
author = {Croce AC and Ferrigno A and Di Pasqua LG and Berardo C and Mannucci B and Bottiroli G and Vairetti M},
url = {http://onlinelibrary.wiley.com/doi/10.1002/jbio.201600195/abstract},
doi = {10.1002/jbio.201600195},
year = {2017},
date = {2017-02-24},
journal = {Journal of biophotonics},
volume = {10},
number = {6-7},
pages = {905-910},
abstract = {The autofluorescence (AF) of NAD(P)H and flavins has been at the basis of many in-situ studies of liver energy metabolism and functionality. Conversely, few data have been so far reported on fluorescing lipids. In this work we investigated the AF of liver lipid extracts from two fatty liver models, Wistar rats fed with MCD diet for 12 days (Wi-MCD), and obese (fa/fa) Zucker rats. Among the most abundant fatty acids in the lipid extracts, indicated by mass spectrometry, arachidonic acid (AA) exhibited higher quantum yield than the other fluorescing fatty acids (FLFA), and red shifted AF spectrum. This allowed to estimate the AA contribution to the overall emission of lipid extracts by curve fitting analysis. AA prevailed in obese Zucker livers, accounting for the different AF spectral profiles between the two models. AF and mass spectrometry indicated also a different balance between the fluorescing fraction and the overall amount of AA in the two models. The ability of AF to detect directly AA and FLFA was demonstrated, suggesting its supportive role as tool in wide-ranging applications, from the control of animal origin food, to experimental investigations on liver fat accumulation, lipotoxicity and disease progression, with potential translation to the clinics.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The autofluorescence (AF) of NAD(P)H and flavins has been at the basis of many in-situ studies of liver energy metabolism and functionality. Conversely, few data have been so far reported on fluorescing lipids. In this work we investigated the AF of liver lipid extracts from two fatty liver models, Wistar rats fed with MCD diet for 12 days (Wi-MCD), and obese (fa/fa) Zucker rats. Among the most abundant fatty acids in the lipid extracts, indicated by mass spectrometry, arachidonic acid (AA) exhibited higher quantum yield than the other fluorescing fatty acids (FLFA), and red shifted AF spectrum. This allowed to estimate the AA contribution to the overall emission of lipid extracts by curve fitting analysis. AA prevailed in obese Zucker livers, accounting for the different AF spectral profiles between the two models. AF and mass spectrometry indicated also a different balance between the fluorescing fraction and the overall amount of AA in the two models. The ability of AF to detect directly AA and FLFA was demonstrated, suggesting its supportive role as tool in wide-ranging applications, from the control of animal origin food, to experimental investigations on liver fat accumulation, lipotoxicity and disease progression, with potential translation to the clinics. |
Croce AC; Ferrigno A; Bertone V; Piccolini VM; Berardo C; Di Pasqua LG; Rizzo V; Bottiroli G; Vairetti M Fatty liver oxidative events monitored by autofluorescence optical diagnosis: a comparison between subnormothermic machine perfusion and conventional cold storage preservation. Journal Article In: Hepatology research, vol. 47, no 7, pp. 668-682, 2017. @article{%a1:%Y_225,
title = {Fatty liver oxidative events monitored by autofluorescence optical diagnosis: a comparison between subnormothermic machine perfusion and conventional cold storage preservation.},
author = {Croce AC and Ferrigno A and Bertone V and Piccolini VM and Berardo C and Di Pasqua LG and Rizzo V and Bottiroli G and Vairetti M},
url = {http://onlinelibrary.wiley.com/doi/10.1111/hepr.12779/epdf},
doi = {10.1111/hepr.12779},
year = {2017},
date = {2017-02-24},
journal = {Hepatology research},
volume = {47},
number = {7},
pages = {668-682},
abstract = {AIM: Livers with moderate steatosis are currently recruited as marginal organs to face donor shortage in transplantation, despite lipid excess and oxidative stress increase preservation injury risk. A sensitive, real time detection of liver metabolism engagement could help donor selection and preservation procedures, ameliorating the graft outcome. Hence, we investigated endogenous biomolecules with autofluorescence (AF) properties as biomarkers supporting the detection of liver oxidative events and the assessment of metabolic responses to external stimuli. METHODS: Livers from male Wistar rats fed with a 12-day methionine-choline deficient (MCD) diet were submitted to AF spectrofluorometric analysis (fiber-optic probe, 366 nm excitation) before and after organ isolation, and following preservation (Cold-Storage-CS, or 20 Celsius degree Machine-Perfusion-MP) and reperfusion. RESULTS: Innovative dynamic AF results on lipid oxidation to lipofuscin-like-lipopigments, correlating with biochemical oxidative damage (TBARS) and antioxidant defense (GSH) parameters, suggested a lipid engagement in MCD livers counteracting reactive oxidizing species. A maintained MCD liver functionality was supported by limited changes in bilirubin AF spectral profile, reflecting bile composition balance, despite their intrinsic mitochondrial weakness was confirmed by ATP levels, and regardless of different CS or MP preservation effects on energy metabolism revealed by conventional NAD(P)H and flavin AF data. CONCLUSION: AF showed that at a relatively short MCD diet time livers are still able to face oxidizing events, maintaining a functional balance. These results strengthen AF as a supportive diagnostic tool in experimental hepatology, to characterize in real time marginal livers, monitor their response to ischemia/reperfusion and to investigate protective therapeutic agents. This article is protected by copyright. All rights reserved.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
AIM: Livers with moderate steatosis are currently recruited as marginal organs to face donor shortage in transplantation, despite lipid excess and oxidative stress increase preservation injury risk. A sensitive, real time detection of liver metabolism engagement could help donor selection and preservation procedures, ameliorating the graft outcome. Hence, we investigated endogenous biomolecules with autofluorescence (AF) properties as biomarkers supporting the detection of liver oxidative events and the assessment of metabolic responses to external stimuli. METHODS: Livers from male Wistar rats fed with a 12-day methionine-choline deficient (MCD) diet were submitted to AF spectrofluorometric analysis (fiber-optic probe, 366 nm excitation) before and after organ isolation, and following preservation (Cold-Storage-CS, or 20 Celsius degree Machine-Perfusion-MP) and reperfusion. RESULTS: Innovative dynamic AF results on lipid oxidation to lipofuscin-like-lipopigments, correlating with biochemical oxidative damage (TBARS) and antioxidant defense (GSH) parameters, suggested a lipid engagement in MCD livers counteracting reactive oxidizing species. A maintained MCD liver functionality was supported by limited changes in bilirubin AF spectral profile, reflecting bile composition balance, despite their intrinsic mitochondrial weakness was confirmed by ATP levels, and regardless of different CS or MP preservation effects on energy metabolism revealed by conventional NAD(P)H and flavin AF data. CONCLUSION: AF showed that at a relatively short MCD diet time livers are still able to face oxidizing events, maintaining a functional balance. These results strengthen AF as a supportive diagnostic tool in experimental hepatology, to characterize in real time marginal livers, monitor their response to ischemia/reperfusion and to investigate protective therapeutic agents. This article is protected by copyright. All rights reserved. |
Garbelli A; Riva V; Crespan E; Maga G How to win the HIV-1 drug resistance hurdle race: running faster or jumping higher? Journal Article In: Biochemical journal, vol. 474, no 10, pp. 1559-1577, 2017. @article{%a1:%Y_219,
title = {How to win the HIV-1 drug resistance hurdle race: running faster or jumping higher?},
author = {Garbelli A and Riva V and Crespan E and Maga G},
url = {http://www.biochemj.org/content/474/10/1559.long},
doi = {10.1042/BCJ20160772},
year = {2017},
date = {2017-02-23},
journal = {Biochemical journal},
volume = {474},
number = {10},
pages = {1559-1577},
abstract = {Infections by the human immunodeficiency virus type 1 (HIV-1), the causative agent of the acquired immunodeficiency syndrome (AIDS), are still totaling an appalling 36.7 millions worldwide, with 1.1 million AIDS deaths/year and a similar number of yearly new infections. All this, in spite of the discovery of HIV-1 as the AIDS etiological agent more than 30 years ago and the introduction of an effective combinatorial antiretroviral therapy (cART), able to control disease progression, more than 20 years ago. Although very effective, current cART is plagued by the emergence of drug-resistant viral variants and most of the efforts in the development of novel direct-acting antiviral agents (DAAs) against HIV-1 have been devoted toward the fighting of resistance. In this review, rather than providing a detailed listing of all the drugs and the corresponding resistance mutations, we aim, through relevant examples, at presenting to the general reader the conceptual shift in the approaches that are being taken to overcome the viral resistance hurdle. From the classic 'running faster' strategy, based on the development of novel DAAs active against the mutant viruses selected by the previous drugs and/or presenting to the virus a high genetic barrier toward the development of resilience, to a 'jumping higher' approach, which looks at the cell, rather than the virus, as a source of valuable drug targets, in order to make the cellular environment non-permissive toward the replication of both wild-type and mutated viruses. 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Infections by the human immunodeficiency virus type 1 (HIV-1), the causative agent of the acquired immunodeficiency syndrome (AIDS), are still totaling an appalling 36.7 millions worldwide, with 1.1 million AIDS deaths/year and a similar number of yearly new infections. All this, in spite of the discovery of HIV-1 as the AIDS etiological agent more than 30 years ago and the introduction of an effective combinatorial antiretroviral therapy (cART), able to control disease progression, more than 20 years ago. Although very effective, current cART is plagued by the emergence of drug-resistant viral variants and most of the efforts in the development of novel direct-acting antiviral agents (DAAs) against HIV-1 have been devoted toward the fighting of resistance. In this review, rather than providing a detailed listing of all the drugs and the corresponding resistance mutations, we aim, through relevant examples, at presenting to the general reader the conceptual shift in the approaches that are being taken to overcome the viral resistance hurdle. From the classic 'running faster' strategy, based on the development of novel DAAs active against the mutant viruses selected by the previous drugs and/or presenting to the virus a high genetic barrier toward the development of resilience, to a 'jumping higher' approach, which looks at the cell, rather than the virus, as a source of valuable drug targets, in order to make the cellular environment non-permissive toward the replication of both wild-type and mutated viruses. 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society. |
Boriani F; Fazio N; Fotia C; Savarino L; Nicoli Aldini N; Martini L; Zini N; Bernardini M; Baldini N A novel technique for decellularization of allogenic nerves and in vivo study of their use for peripheral nerve reconstruction. Journal Article In: Journal of biomedical materials research. Part A, vol. 105, no 8, pp. 2228-2240, 2017. @article{%a1:%Y_226,
title = {A novel technique for decellularization of allogenic nerves and in vivo study of their use for peripheral nerve reconstruction.},
author = {Boriani F and Fazio N and Fotia C and Savarino L and Nicoli Aldini N and Martini L and Zini N and Bernardini M and Baldini N},
url = {http://onlinelibrary.wiley.com/doi/10.1002/jbm.a.36090/abstract},
doi = {10.1002/jbm.a.36090},
year = {2017},
date = {2017-02-23},
journal = {Journal of biomedical materials research. Part A},
volume = {105},
number = {8},
pages = {2228-2240},
abstract = {Autografts represent the gold standard for peripheral nerve reconstruction but their limited availability, the discrepancy of nerve caliber and long surgical times are drawbacks. Allografts have therefore become a valid alternative option. In particular, acellular nerve allografts (ANAs) rather than fresh allografts do not need immunosuppression and appear to be safe and effective based on recent studies. An innovative method was conceived to obtain ANAs, so as to speed up nerve decellularization, without compromising nerve architecture, and without breaking the asepsis chain. Several detergent-based techniques, integrated with sonication and mechanical stirring, were tested in vitro on rabbit nerves, to identify, by microscopy and immunohistochemistry, the most effective protocol in terms of cell lysis and cellular debris clearance, while maintaining nerve architecture. Furthermore, a pilot in vivo study was performed: ANAs were implanted into tibial nerve defects of three rabbits, and autografts, representing the gold standard, in other three animals. Twelve weeks postoperatively, rabbits were clinically evaluated and euthanasized; grafts were harvested and microscopically and histomorphometrically analyzed. The method proved to be effective in vitro: the treatment removed axons, myelin and cells, without altering nerve architecture. The in vivo study did not reveal any adverse effect: animals maintained normal weight and function of posterior limb during the entire experimental time. A mild fibrotic reaction was observed, macrophages and leukocytes were rare or absent; ANAs regenerated fascicles and bundles were comparable versus autografts. Based on these results, this decellularization protocol is encouraging and deserves deeper investigations with further pre-clinical and clinical studies. This article is protected by copyright. All rights reserved.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Autografts represent the gold standard for peripheral nerve reconstruction but their limited availability, the discrepancy of nerve caliber and long surgical times are drawbacks. Allografts have therefore become a valid alternative option. In particular, acellular nerve allografts (ANAs) rather than fresh allografts do not need immunosuppression and appear to be safe and effective based on recent studies. An innovative method was conceived to obtain ANAs, so as to speed up nerve decellularization, without compromising nerve architecture, and without breaking the asepsis chain. Several detergent-based techniques, integrated with sonication and mechanical stirring, were tested in vitro on rabbit nerves, to identify, by microscopy and immunohistochemistry, the most effective protocol in terms of cell lysis and cellular debris clearance, while maintaining nerve architecture. Furthermore, a pilot in vivo study was performed: ANAs were implanted into tibial nerve defects of three rabbits, and autografts, representing the gold standard, in other three animals. Twelve weeks postoperatively, rabbits were clinically evaluated and euthanasized; grafts were harvested and microscopically and histomorphometrically analyzed. The method proved to be effective in vitro: the treatment removed axons, myelin and cells, without altering nerve architecture. The in vivo study did not reveal any adverse effect: animals maintained normal weight and function of posterior limb during the entire experimental time. A mild fibrotic reaction was observed, macrophages and leukocytes were rare or absent; ANAs regenerated fascicles and bundles were comparable versus autografts. Based on these results, this decellularization protocol is encouraging and deserves deeper investigations with further pre-clinical and clinical studies. This article is protected by copyright. All rights reserved. |
Iannelli F; Galbiati A; Capozzo I; Nguyen Q; Magnuson B; Michelini F; D'Alessandro G; Cabrini M; Roncador M; Francia S; Crosetto N; Ljungman M; Carninci P; d'Adda di Fagagna F A damaged genome’s transcriptional landscape through multilayered expression profiling around in situ-mapped DNA double-strand breaks Journal Article In: Nature Communications, vol. 8, pp. 15656, 2017. @article{%a1:%Y_185,
title = {A damaged genome’s transcriptional landscape through multilayered expression profiling around in situ-mapped DNA double-strand breaks},
author = {Iannelli F and Galbiati A and Capozzo I and Nguyen Q and Magnuson B and Michelini F and D'Alessandro G and Cabrini M and Roncador M and Francia S and Crosetto N and Ljungman M and Carninci P and {d'Adda di Fagagna F}},
url = {https://www.nature.com/articles/ncomms15656},
doi = {10.1038/ncomms15656},
year = {2017},
date = {2017-02-23},
journal = {Nature Communications},
volume = {8},
pages = {15656},
abstract = {Of the many types of DNA damage, DNA double-strand breaks (DSBs) are probably the most deleterious. Mounting evidence points to an intricate relationship between DSBs and transcription. A cell system in which the impact on transcription can be investigated at precisely mapped genomic DSBs is essential to study this relationship. Here in a human cell line, we map genome-wide and at high resolution the DSBs induced by a restriction enzyme, and we characterize their impact on gene expression by four independent approaches by monitoring steady-state RNA levels, rates of RNA synthesis, transcription initiation and RNA polymerase II elongation. We consistently observe transcriptional repression in proximity to DSBs. Downregulation of transcription depends on ATM kinase activity and on the distance from the DSB. Our study couples for the first time, to the best of our knowledge, high-resolution mapping of DSBs with multilayered transcriptomics to dissect the events shaping gene expression after DSB induction at multiple endogenous sites.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Of the many types of DNA damage, DNA double-strand breaks (DSBs) are probably the most deleterious. Mounting evidence points to an intricate relationship between DSBs and transcription. A cell system in which the impact on transcription can be investigated at precisely mapped genomic DSBs is essential to study this relationship. Here in a human cell line, we map genome-wide and at high resolution the DSBs induced by a restriction enzyme, and we characterize their impact on gene expression by four independent approaches by monitoring steady-state RNA levels, rates of RNA synthesis, transcription initiation and RNA polymerase II elongation. We consistently observe transcriptional repression in proximity to DSBs. Downregulation of transcription depends on ATM kinase activity and on the distance from the DSB. Our study couples for the first time, to the best of our knowledge, high-resolution mapping of DSBs with multilayered transcriptomics to dissect the events shaping gene expression after DSB induction at multiple endogenous sites. |
Famiglini V; La Regina G; Coluccia A; Masci D; Brancale A; Badia R; Riveira-Munoz E; Esté JA; Crespan E; Brambilla A; Maga G; Catalano M; Limatola C; Formica FR; Cirilli R; Novellino E; Silvestri R Chiral Indolylarylsulfone Non-Nucleoside Reverse Transcriptase Inhibitors as New Potent and Broad Spectrum Anti-HIV-1 Agents. Journal Article In: Journal of medicinal chemistry, vol. 60, no 15, pp. 6528-6547, 2017. @article{%a1:%Y_198,
title = {Chiral Indolylarylsulfone Non-Nucleoside Reverse Transcriptase Inhibitors as New Potent and Broad Spectrum Anti-HIV-1 Agents.},
author = {Famiglini V and La Regina G and Coluccia A and Masci D and Brancale A and Badia R and Riveira-Munoz E and Esté JA and Crespan E and Brambilla A and Maga G and Catalano M and Limatola C and Formica FR and Cirilli R and Novellino E and Silvestri R},
url = {http://pubs.acs.org/doi/10.1021/acs.jmedchem.6b01906},
doi = {10.1021/acs.jmedchem.6b01906},
year = {2017},
date = {2017-02-23},
journal = {Journal of medicinal chemistry},
volume = {60},
number = {15},
pages = {6528-6547},
abstract = {We designed and synthesized a series of chiral indolyarylsulfones (IASs) as new HIV-1 NNRTIs. The new IASs 8-37 showed potent inhibition of the HIV-1 WT NL4-3 strain and of the mutant K103N, Y181C, Y188L, and K103N-Y181C HIV-1 strains. Six racemic mixtures, 8, 23-25, 31, and 33, were separated at semipreparative level into their pure enantiomers. The (R)-8 enantiomer bearing the chiral (α-methylbenzyl) was superior to the (S)-counterpart. IAS derivatives bearing the (S) alanine unit, (S)-23, (S,R)-25, (S)-31, and (S)-33, were remarkably more potent than the corresponding (R)-enantiomers. Compound 23 protected hippocampal neuronal cells from the excitotoxic insult, while efavirenz (EFV) did not contrast the neurotoxic effect of glutamate. The present results highlight the chiral IASs as new NNRTIs with improved resistance profile against the mutant HIV-1 strains and reduced neurotoxic effects.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
We designed and synthesized a series of chiral indolyarylsulfones (IASs) as new HIV-1 NNRTIs. The new IASs 8-37 showed potent inhibition of the HIV-1 WT NL4-3 strain and of the mutant K103N, Y181C, Y188L, and K103N-Y181C HIV-1 strains. Six racemic mixtures, 8, 23-25, 31, and 33, were separated at semipreparative level into their pure enantiomers. The (R)-8 enantiomer bearing the chiral (α-methylbenzyl) was superior to the (S)-counterpart. IAS derivatives bearing the (S) alanine unit, (S)-23, (S,R)-25, (S)-31, and (S)-33, were remarkably more potent than the corresponding (R)-enantiomers. Compound 23 protected hippocampal neuronal cells from the excitotoxic insult, while efavirenz (EFV) did not contrast the neurotoxic effect of glutamate. The present results highlight the chiral IASs as new NNRTIs with improved resistance profile against the mutant HIV-1 strains and reduced neurotoxic effects. |
Dutto I; Cazzalini O; Stivala LA; Prosperi E An improved method for the detection of nucleotide excision repair factors at local UV DNA damage sites. Journal Article In: DNA Repair, vol. 51, pp. 79-84, 2017. @article{%a1:%Y_211,
title = {An improved method for the detection of nucleotide excision repair factors at local UV DNA damage sites.},
author = {Dutto I and Cazzalini O and Stivala LA and Prosperi E},
url = {http://www.sciencedirect.com/science/article/pii/S1568786417300034},
doi = {dx.doi.org/10.1016/j.dnarep.2017.01.005},
year = {2017},
date = {2017-02-22},
journal = {DNA Repair},
volume = {51},
pages = {79-84},
abstract = {Among different DNA repair processes that cells use to face with DNA damage, nucleotide excision repair (NER) is particularly important for the removal of a high variety of lesions, including those generated by some antitumor drugs. A number of factors participating in NER, such as the TFIIH complex and the endonuclease XPG are also involved in basal processes, e.g. transcription. For this reason, localization of these factors at DNA damage sites may be difficult. Here we have applied a mild digestion of chromatin with DNase I to improve the in situ extraction necessary to detect chromatin-bound proteins by immunofluorescence. We have compared this method with different extraction protocols and investigated its application on different cell types, and with different antibodies. Our results show that a short DNase I treatment before the immunoreaction, enhances the fluorescence signal of NER proteins, such as XPG, DDB2 and XPC. In addition, our findings indicate that the antibody choice is a critical factor for accurate localization of DNA repair proteins at DNA damage sites. In conclusion, a mild DNA digestion with DNase I improves the immunofluorescence detection of the recruitment of NER factors at local DNA damage sites by enhancing accessibility to the antibodies, independently of the cell type.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Among different DNA repair processes that cells use to face with DNA damage, nucleotide excision repair (NER) is particularly important for the removal of a high variety of lesions, including those generated by some antitumor drugs. A number of factors participating in NER, such as the TFIIH complex and the endonuclease XPG are also involved in basal processes, e.g. transcription. For this reason, localization of these factors at DNA damage sites may be difficult. Here we have applied a mild digestion of chromatin with DNase I to improve the in situ extraction necessary to detect chromatin-bound proteins by immunofluorescence. We have compared this method with different extraction protocols and investigated its application on different cell types, and with different antibodies. Our results show that a short DNase I treatment before the immunoreaction, enhances the fluorescence signal of NER proteins, such as XPG, DDB2 and XPC. In addition, our findings indicate that the antibody choice is a critical factor for accurate localization of DNA repair proteins at DNA damage sites. In conclusion, a mild DNA digestion with DNase I improves the immunofluorescence detection of the recruitment of NER factors at local DNA damage sites by enhancing accessibility to the antibodies, independently of the cell type. |
Pignataro D; Francia S; Zanetta F; Brenna G; Brandini S; Olivieri A; Torroni A; Biamonti G; Montecucco A A missense MT-ND5 mutation in differentiated Parkinson Disease cytoplasmic hybrid induces ROS-dependent DNA Damage Response amplified by DROSHA. Journal Article In: Scientific reports, vol. 7, no 1, pp. 9528, 2017. @article{%a1:%Y_214,
title = {A missense MT-ND5 mutation in differentiated Parkinson Disease cytoplasmic hybrid induces ROS-dependent DNA Damage Response amplified by DROSHA.},
author = {Pignataro D and Francia S and Zanetta F and Brenna G and Brandini S and Olivieri A and Torroni A and Biamonti G and Montecucco A},
url = {https://www.nature.com/articles/s41598-017-09910-x},
doi = {10.1038/s41598-017-09910-x},
year = {2017},
date = {2017-02-22},
journal = {Scientific reports},
volume = {7},
number = {1},
pages = {9528},
abstract = {Genome integrity is continuously threatened by endogenous sources of DNA damage including reactive oxygen species (ROS) produced by cell metabolism. Factors of the RNA interference (RNAi) machinery have been recently involved in the cellular response to DNA damage (DDR) in proliferating cells. To investigate the impact of component of RNAi machinery on DDR activation in terminally differentiated cells, we exploited cytoplasmic hybrid (cybrid) cell lines in which mitochondria of sporadic Parkinson's disease patients repopulate neuroblastoma SH-SY5Y-Rho(0) cells. Upon differentiation into dopaminergic neuron-like cells, PD63 cybrid showed increased intracellular level of ROS and chronic DDR activation, compared to other cybrids with the same nuclear background. Importantly, DDR activation in these cells can be prevented by ROS scavenging treatment suggesting that ROS production is indeed causative of nuclear DNA damage. Sequence analysis of the mitogenomes identified a rare and heteroplasmic missense mutation affecting a highly conserved residue of the ND5-subunit of respiratory complex I, which accounts for ROS increase. We demonstrated that the assembly of nuclear DDR foci elicited by oxidative stress in these cells relies on DROSHA, providing the first evidence that components of RNAi machinery play a crucial role also in the mounting of ROS-induced DDR in non-replicating neuronal cells.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Genome integrity is continuously threatened by endogenous sources of DNA damage including reactive oxygen species (ROS) produced by cell metabolism. Factors of the RNA interference (RNAi) machinery have been recently involved in the cellular response to DNA damage (DDR) in proliferating cells. To investigate the impact of component of RNAi machinery on DDR activation in terminally differentiated cells, we exploited cytoplasmic hybrid (cybrid) cell lines in which mitochondria of sporadic Parkinson's disease patients repopulate neuroblastoma SH-SY5Y-Rho(0) cells. Upon differentiation into dopaminergic neuron-like cells, PD63 cybrid showed increased intracellular level of ROS and chronic DDR activation, compared to other cybrids with the same nuclear background. Importantly, DDR activation in these cells can be prevented by ROS scavenging treatment suggesting that ROS production is indeed causative of nuclear DNA damage. Sequence analysis of the mitogenomes identified a rare and heteroplasmic missense mutation affecting a highly conserved residue of the ND5-subunit of respiratory complex I, which accounts for ROS increase. We demonstrated that the assembly of nuclear DDR foci elicited by oxidative stress in these cells relies on DROSHA, providing the first evidence that components of RNAi machinery play a crucial role also in the mounting of ROS-induced DDR in non-replicating neuronal cells. |
Michelini F; Pitchiaya S; Vitelli V; Sharma S; Gioia U; Pessina F; Cabrini M; Wang Y; Capozzo I; Iannelli F; Matti V; Francia S; Shivashankar GV; Walter NG; d'Adda di Fagagna F Damage-induced lncRNAs control the DNA damage response through interaction with DDRNAs at individual double-strand breaks Journal Article In: Nature cell biology, vol. 19, iss. 12, pp. 1400-1410, 2017. @article{%a1:%Y_184,
title = {Damage-induced lncRNAs control the DNA damage response through interaction with DDRNAs at individual double-strand breaks},
author = {Michelini F and Pitchiaya S and Vitelli V and Sharma S and Gioia U and Pessina F and Cabrini M and Wang Y and Capozzo I and Iannelli F and Matti V and Francia S and Shivashankar GV and Walter NG and {d'Adda di Fagagna F}},
url = {https://www.nature.com/articles/ncb3643},
doi = {10.1038/ncb3643},
year = {2017},
date = {2017-02-22},
journal = {Nature cell biology},
volume = {19},
issue = {12},
pages = {1400-1410},
abstract = {The DNA damage response (DDR) preserves genomic integrity. Small non-coding RNAs termed DDRNAs are generated at DNA double-strand breaks (DSBs) and are critical for DDR activation. Here we show that active DDRNAs specifically localize to their damaged homologous genomic sites in a transcription-dependent manner. Following DNA damage, RNA polymerase II (RNAPII) binds to the MRE11-RAD50-NBS1 complex, is recruited to DSBs and synthesizes damage-induced long non-coding RNAs (dilncRNAs) from and towards DNA ends. DilncRNAs act both as DDRNA precursors and by recruiting DDRNAs through RNA-RNA pairing. Together, dilncRNAs and DDRNAs fuel DDR focus formation and associate with 53BP1. Accordingly, inhibition of RNAPII prevents DDRNA recruitment, DDR activation and DNA repair. Antisense oligonucleotides matching dilncRNAs and DDRNAs impair site-specific DDR focus formation and DNA repair. We propose that DDR signalling sites, in addition to sharing a common pool of proteins, individually host a unique set of site-specific RNAs necessary for DDR activation.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The DNA damage response (DDR) preserves genomic integrity. Small non-coding RNAs termed DDRNAs are generated at DNA double-strand breaks (DSBs) and are critical for DDR activation. Here we show that active DDRNAs specifically localize to their damaged homologous genomic sites in a transcription-dependent manner. Following DNA damage, RNA polymerase II (RNAPII) binds to the MRE11-RAD50-NBS1 complex, is recruited to DSBs and synthesizes damage-induced long non-coding RNAs (dilncRNAs) from and towards DNA ends. DilncRNAs act both as DDRNA precursors and by recruiting DDRNAs through RNA-RNA pairing. Together, dilncRNAs and DDRNAs fuel DDR focus formation and associate with 53BP1. Accordingly, inhibition of RNAPII prevents DDRNA recruitment, DDR activation and DNA repair. Antisense oligonucleotides matching dilncRNAs and DDRNAs impair site-specific DDR focus formation and DNA repair. We propose that DDR signalling sites, in addition to sharing a common pool of proteins, individually host a unique set of site-specific RNAs necessary for DDR activation. |
Aredia F; Scovassi AI A new function for miRNAs as regulators of autophagy. Journal Article In: Future Medicine Chemistry, vol. 9, no 1, pp. 25-36, 2017. @article{%a1:%Y_217,
title = {A new function for miRNAs as regulators of autophagy.},
author = {Aredia F and Scovassi AI},
url = {http://www.future-science.com/doi/abs/10.4155/fmc-2016-0173?url_ver=Z39.88-2003&rfr_id=ori%3Arid%3Acrossref.org&rfr_dat=cr_pub%3Dpubmed&},
doi = {10.4155/fmc-2016-0173},
year = {2017},
date = {2017-02-17},
journal = {Future Medicine Chemistry},
volume = {9},
number = {1},
pages = {25-36},
abstract = {Autophagy is a self-digestive process regulated by an intricate network of factors able either to ensure the prosurvival function of autophagy or to convert it in a death pathway. Recently, the involvement of miRNAs in the regulation of autophagy networks has been reported. This review will summarize the main features of these small noncoding endogenous RNAs, focusing on their relevance in cancer and finally addressing their impact on autophagy.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Autophagy is a self-digestive process regulated by an intricate network of factors able either to ensure the prosurvival function of autophagy or to convert it in a death pathway. Recently, the involvement of miRNAs in the regulation of autophagy networks has been reported. This review will summarize the main features of these small noncoding endogenous RNAs, focusing on their relevance in cancer and finally addressing their impact on autophagy. |
Croce AC; Bottiroli G Lipids: Evergreen autofluorescent biomarkers for the liver functional profiling. Journal Article In: European Journal of Histochemistry, vol. 61, no 2, pp. 2808, 2017. @article{%a1:%Y_229,
title = {Lipids: Evergreen autofluorescent biomarkers for the liver functional profiling.},
author = {Croce AC and Bottiroli G},
url = {http://ejh.it/index.php/ejh/article/view/2808},
doi = {10.4081/ejh.2017.2808},
year = {2017},
date = {2017-02-17},
journal = {European Journal of Histochemistry},
volume = {61},
number = {2},
pages = {2808},
abstract = {Depending on their chemical nature, lipids can be classified in two main categories: hydrophilic, greatly contributing to membrane composition and subcellular organelle compartmentalization, and hydrophobic, mostly triglycerides, greatly enrolled in the storage and production of energy. In both cases, some lipid molecules can be involved as signaling agents in the regulation of metabolism and protective or damaging pathways in responses to harmful stimuli. These events could affect in particular the liver, because of its central role in the maintenance of lipid homeostasis. Lipids have been demonstrated to fluoresce, contributing to the overall emission signal of the liver tissue along with other endogenous fluorophores, relatable to energy metabolism and oxidative events. The mere estimation of the fluorescing lipid fraction in parallel with the other endogenous fluorophores, and with the common biochemical and histochemical biomarkers of tissue injury has been exploited to investigate the liver morpho-functional conditions in experimental hepatology. More interestingly, the fluorescing lipid fraction is greatly relatable to free fatty acids such as arachidonic, linoleic and linolenic acid, which are deserving increasing attention as precursors of products involved in several and complex signaling pathways. On these bases, the ability of autofluorescence to detect directly arachidonic acid and its balance with other unsaturated fatty acids may be exploited in the diagnosis and follow-up of fatty livers, helping to improve the personalization of the metabolic/lipidomic profiling. This could also contribute to elucidate the role of the injuring factors in the choice of suitable donors, and in the set-up of preservation procedures in liver transplantation.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Depending on their chemical nature, lipids can be classified in two main categories: hydrophilic, greatly contributing to membrane composition and subcellular organelle compartmentalization, and hydrophobic, mostly triglycerides, greatly enrolled in the storage and production of energy. In both cases, some lipid molecules can be involved as signaling agents in the regulation of metabolism and protective or damaging pathways in responses to harmful stimuli. These events could affect in particular the liver, because of its central role in the maintenance of lipid homeostasis. Lipids have been demonstrated to fluoresce, contributing to the overall emission signal of the liver tissue along with other endogenous fluorophores, relatable to energy metabolism and oxidative events. The mere estimation of the fluorescing lipid fraction in parallel with the other endogenous fluorophores, and with the common biochemical and histochemical biomarkers of tissue injury has been exploited to investigate the liver morpho-functional conditions in experimental hepatology. More interestingly, the fluorescing lipid fraction is greatly relatable to free fatty acids such as arachidonic, linoleic and linolenic acid, which are deserving increasing attention as precursors of products involved in several and complex signaling pathways. On these bases, the ability of autofluorescence to detect directly arachidonic acid and its balance with other unsaturated fatty acids may be exploited in the diagnosis and follow-up of fatty livers, helping to improve the personalization of the metabolic/lipidomic profiling. This could also contribute to elucidate the role of the injuring factors in the choice of suitable donors, and in the set-up of preservation procedures in liver transplantation. |
Croce AC; Ferrigno A; Di Pasqua LG; Berardo C; Bottiroli G; Vairetti M NAD(P)H and Flavin Autofluorescence Correlation with ATP in Rat Livers with Different Metabolic Steady-State Conditions. Journal Article In: Photochemistry and photobiology, vol. 93, no 6, pp. 1519-1524, 2017. @article{%a1:%Y_230,
title = {NAD(P)H and Flavin Autofluorescence Correlation with ATP in Rat Livers with Different Metabolic Steady-State Conditions.},
author = {Croce AC and Ferrigno A and Di Pasqua LG and Berardo C and Bottiroli G and Vairetti M},
url = {http://onlinelibrary.wiley.com/doi/10.1111/php.12804/abstract},
doi = {10.1111/php.12804},
year = {2017},
date = {2017-02-17},
journal = {Photochemistry and photobiology},
volume = {93},
number = {6},
pages = {1519-1524},
abstract = {The monitoring of NAD(P)H and flavin autofluorescence (AF) is at the basis of numerous investigations on energy metabolism. Nevertheless, the ability of these AF biomarkers to accurately represent the energy currency, ATP, is poorly explored. Here, we focused on the AF/ATP correlation in lean and fatty livers with different steady-state metabolic conditions, achieved after organ isolation, preservation and recovery, in a likely dependence on both liver intrinsic metabolic features and externally induced perturbations. Within these eventual, various conditions, a significant correlation was detected between liver NAD(P)H and flavin AF, measured via fiber-optic probe, and biochemical ATP data, strengthening AF as biomarker of energy metabolism in steady-state conditions for wide-ranging experimental and diagnostic applications.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The monitoring of NAD(P)H and flavin autofluorescence (AF) is at the basis of numerous investigations on energy metabolism. Nevertheless, the ability of these AF biomarkers to accurately represent the energy currency, ATP, is poorly explored. Here, we focused on the AF/ATP correlation in lean and fatty livers with different steady-state metabolic conditions, achieved after organ isolation, preservation and recovery, in a likely dependence on both liver intrinsic metabolic features and externally induced perturbations. Within these eventual, various conditions, a significant correlation was detected between liver NAD(P)H and flavin AF, measured via fiber-optic probe, and biochemical ATP data, strengthening AF as biomarker of energy metabolism in steady-state conditions for wide-ranging experimental and diagnostic applications. |
Duchi S; Piccinini F; Pierini M; Bevilacqua A; Torre ML; Lucarelli E; Santi S A new holistic 3D non-invasive analysis of cellular distribution and motility on fibroin-alginate microcarriers using light sheet fluorescent microscopy. Journal Article In: Plos One, vol. 12, no 8, pp. e0183336, 2017. @article{%a1:%Y_227,
title = {A new holistic 3D non-invasive analysis of cellular distribution and motility on fibroin-alginate microcarriers using light sheet fluorescent microscopy.},
author = {Duchi S and Piccinini F and Pierini M and Bevilacqua A and Torre ML and Lucarelli E and Santi S},
url = {http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0183336},
doi = {0.1371/journal.pone.0183336},
year = {2017},
date = {2017-02-16},
journal = {Plos One},
volume = {12},
number = {8},
pages = {e0183336},
abstract = {Cell interaction with biomaterials is one of the keystones to developing medical devices for tissue engineering applications. Biomaterials are the scaffolds that give three-dimensional support to the cells, and are vectors that deliver the cells to the injured tissue requiring repair. Features of biomaterials can influence the behaviour of the cells and consequently the efficacy of the tissue-engineered product. The adhesion, distribution and motility of the seeded cells onto the scaffold represent key aspects, and must be evaluated in vitro during the product development, especially when the efficacy of a specific tissue-engineered product depends on viable and functional cell loading. In this work, we propose a non-invasive and non-destructive imaging analysis for investigating motility, viability and distribution of Mesenchymal Stem Cells (MSCs) on silk fibroin-based alginate microcarriers, to test the adhesion capacity of the fibroin coating onto alginate which is known to be unsuitable for cell adhesion. However, in depth characterization of the biomaterial is beyond the scope of this paper. Scaffold-loaded MSCs were stained with Calcein-AM and Ethidium homodimer-1 to detect live and dead cells, respectively, and counterstained with Hoechst to label cell nuclei. Time-lapse Light Sheet Fluorescent Microscopy (LSFM) was then used to produce three-dimensional images of the entire cells-loaded fibroin/alginate microcarriers. In order to quantitatively track the cell motility over time, we also developed an open source user friendly software tool called Fluorescent Cell Tracker in Three-Dimensions (F-Tracker3D). Combining LSFM with F-Tracker3D we were able for the first time to assess the distribution and motility of stem cells in a non-invasive, non-destructive, quantitative, and three-dimensional analysis of the entire surface of the cell-loaded scaffold. We therefore propose this imaging technique as an innovative holistic tool for monitoring cell-biomaterial interactions, and as a tool for the design, fabrication and functionalization of a scaffold as a medical device.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Cell interaction with biomaterials is one of the keystones to developing medical devices for tissue engineering applications. Biomaterials are the scaffolds that give three-dimensional support to the cells, and are vectors that deliver the cells to the injured tissue requiring repair. Features of biomaterials can influence the behaviour of the cells and consequently the efficacy of the tissue-engineered product. The adhesion, distribution and motility of the seeded cells onto the scaffold represent key aspects, and must be evaluated in vitro during the product development, especially when the efficacy of a specific tissue-engineered product depends on viable and functional cell loading. In this work, we propose a non-invasive and non-destructive imaging analysis for investigating motility, viability and distribution of Mesenchymal Stem Cells (MSCs) on silk fibroin-based alginate microcarriers, to test the adhesion capacity of the fibroin coating onto alginate which is known to be unsuitable for cell adhesion. However, in depth characterization of the biomaterial is beyond the scope of this paper. Scaffold-loaded MSCs were stained with Calcein-AM and Ethidium homodimer-1 to detect live and dead cells, respectively, and counterstained with Hoechst to label cell nuclei. Time-lapse Light Sheet Fluorescent Microscopy (LSFM) was then used to produce three-dimensional images of the entire cells-loaded fibroin/alginate microcarriers. In order to quantitatively track the cell motility over time, we also developed an open source user friendly software tool called Fluorescent Cell Tracker in Three-Dimensions (F-Tracker3D). Combining LSFM with F-Tracker3D we were able for the first time to assess the distribution and motility of stem cells in a non-invasive, non-destructive, quantitative, and three-dimensional analysis of the entire surface of the cell-loaded scaffold. We therefore propose this imaging technique as an innovative holistic tool for monitoring cell-biomaterial interactions, and as a tool for the design, fabrication and functionalization of a scaffold as a medical device. |
Apolonio FM; Mazzoni A; Angeloni V; Scaffa PM; Santi S; Saboia VP; Tay FR; Pashley DH; Breschi L Effect of a one-step self-etch adhesive on endogenous dentin matrix metalloproteinases. Journal Article In: European journal of oral sciences, vol. 125, no 2, pp. 168-172, 2017. @article{%a1:%Y_233,
title = {Effect of a one-step self-etch adhesive on endogenous dentin matrix metalloproteinases.},
author = {Apolonio FM and Mazzoni A and Angeloni V and Scaffa PM and Santi S and Saboia VP and Tay FR and Pashley DH and Breschi L},
url = {http://onlinelibrary.wiley.com/doi/10.1111/eos.12337/abstract;jsessionid=518F5CAC0BCA0ADC60A80D53F30477C5.f02t03},
doi = {10.1111/eos.12337},
year = {2017},
date = {2017-02-16},
journal = {European journal of oral sciences},
volume = {125},
number = {2},
pages = {168-172},
abstract = {Degradation of the hybrid layer created in dentin by dentin adhesives is caused by enzyme activities present within the dentin matrix that destroy unprotected collagen fibrils. The aim of the present study was to evaluate the effect of a one-step self-etch adhesive system on dentinal matrix metalloproteinases 2 and 4 (MMP-2 and MMP-9, respectively) using in situ zymography and an enzymatic activity assay. The null hypothesis tested was that there are no differences in the activities of dentinal MMPs before and after treatment with a one-step adhesive system. The MMP-2 and MMP-9 activities in dentin treated with the one-step adhesive, Adper Easy Bond, were quantified using an enzymatic activity assay system. The MMP activities within the hybrid layer created by the one-step adhesive tested were also evaluated using in situ zymography. The enzymatic assay revealed an increase in MMP-2 and MMP-9 activities after treatment with adhesive. In situ zymography indicated that gelatinolytic activity is present within the hybrid layer created with the one-step self-etch adhesive. The host-derived gelatinases were localized within the hybrid layer and remained active after the bonding procedure. It is concluded that the one-step self-etch adhesive investigated activates endogenous MMP-2 and MMP-9 with the dentin matrix, which may cause collagen degradation over time.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Degradation of the hybrid layer created in dentin by dentin adhesives is caused by enzyme activities present within the dentin matrix that destroy unprotected collagen fibrils. The aim of the present study was to evaluate the effect of a one-step self-etch adhesive system on dentinal matrix metalloproteinases 2 and 4 (MMP-2 and MMP-9, respectively) using in situ zymography and an enzymatic activity assay. The null hypothesis tested was that there are no differences in the activities of dentinal MMPs before and after treatment with a one-step adhesive system. The MMP-2 and MMP-9 activities in dentin treated with the one-step adhesive, Adper Easy Bond, were quantified using an enzymatic activity assay system. The MMP activities within the hybrid layer created by the one-step adhesive tested were also evaluated using in situ zymography. The enzymatic assay revealed an increase in MMP-2 and MMP-9 activities after treatment with adhesive. In situ zymography indicated that gelatinolytic activity is present within the hybrid layer created with the one-step self-etch adhesive. The host-derived gelatinases were localized within the hybrid layer and remained active after the bonding procedure. It is concluded that the one-step self-etch adhesive investigated activates endogenous MMP-2 and MMP-9 with the dentin matrix, which may cause collagen degradation over time. |
Toniato E; Frydas I; Robuffo I; Ronconi G; Caraffa Al; Kritas SK; Conti P Activation and inhibition of adaptive immune response mediated by mast cells. Journal Article In: Journal of biological regulators and homeostatic agents, vol. 31, no 3, pp. 543-548, 2017. @article{%a1:%Y_234,
title = {Activation and inhibition of adaptive immune response mediated by mast cells.},
author = {Toniato E and Frydas I and Robuffo I and Ronconi G and Caraffa Al and Kritas SK and Conti P},
url = {https://www.biolifesas.org/biolife/category/journals/journal-of-biological-regulators-and-homeostatic-agents/},
year = {2017},
date = {2017-02-16},
journal = {Journal of biological regulators and homeostatic agents},
volume = {31},
number = {3},
pages = {543-548},
abstract = {Adaptive immune response plays an important role against bacteria and parasites, a reaction that also involves mast cell (MC) activation which participates in innate and adaptive immunity. In allergic reactions there is a TH2 immune response with generation of allergen-specific IgE antibodies. In MCs, IgE cross-link FcRI high affinity receptor and activate tyrosine kinase proteins, leading to stimulation of NF-κB and AP-1 resulting in the release of a number of cytokines/chemokines and other compounds. Through their proteolytic pathways, MCs may process the antigen for presentation to CD4+ cells which release TH2 cytokines and growth factors, which play an important role in asthma, allergy, anaphylaxis and inflammation. Thus, MCs can contribute to adaptive immunity. MCs may also be activated though the TLR-dependent pathway which is controlled by several proteins including myeloid differentiation factor 88 (MyD88) which can be inhibited by interleukin (IL)-37. Here, we describe the participation of MCs in adaptive immunity and inflammation, an effect that may be inhibited by IL-37.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Adaptive immune response plays an important role against bacteria and parasites, a reaction that also involves mast cell (MC) activation which participates in innate and adaptive immunity. In allergic reactions there is a TH2 immune response with generation of allergen-specific IgE antibodies. In MCs, IgE cross-link FcRI high affinity receptor and activate tyrosine kinase proteins, leading to stimulation of NF-κB and AP-1 resulting in the release of a number of cytokines/chemokines and other compounds. Through their proteolytic pathways, MCs may process the antigen for presentation to CD4+ cells which release TH2 cytokines and growth factors, which play an important role in asthma, allergy, anaphylaxis and inflammation. Thus, MCs can contribute to adaptive immunity. MCs may also be activated though the TLR-dependent pathway which is controlled by several proteins including myeloid differentiation factor 88 (MyD88) which can be inhibited by interleukin (IL)-37. Here, we describe the participation of MCs in adaptive immunity and inflammation, an effect that may be inhibited by IL-37. |
Ferrigno A; Di Pasqua LG; Berardo C; Siciliano V; Rizzo V; Mannucci B; Richelmi P; Croce AC; Vairetti M Liver Graft Susceptibility during Static Cold Storage and Dynamic Machine Perfusion: DCD versus Fatty Livers. Journal Article In: International journal of molecular sciences, vol. 19, no 1, pp. 109, 2017. @article{%a1:%Y_137,
title = {Liver Graft Susceptibility during Static Cold Storage and Dynamic Machine Perfusion: DCD versus Fatty Livers.},
author = {Ferrigno A and Di Pasqua LG and Berardo C and Siciliano V and Rizzo V and Mannucci B and Richelmi P and Croce AC and Vairetti M},
url = {10.3390/ijms19010109},
doi = {10.3390/ijms19010109},
year = {2017},
date = {2017-02-16},
urldate = {2017-02-16},
journal = {International journal of molecular sciences},
volume = {19},
number = {1},
pages = {109},
abstract = {We compared static preservation (cold storage, CS, 4 C) with dynamic preservation (machine perfusion, MP, 20 C) followed by reperfusion using marginal livers: a model of donation after cardiac death (DCD) livers and two models of fatty livers, the methionine-choline deficient (MCD) diet model, and obese Zucker (fa/fa) rats. CS injury in DCD livers was reversed by an oxygenated washout (OW): hepatic damage, bile flow, and the ATP/ADP ratio in the OW + CS group was comparable with the ratio obtained with MP. Using fatty livers, CS preservation induced a marked release in hepatic and biliary enzymes in obese Zucker rats when compared with the MCD group. The same trend occurred for bile flow. No difference was found when comparing MP in MCD and obese Zucker rats. Fatty acid analysis demonstrated that the total saturated (SFA)/polyunsaturated fatty acid (PUFA) ratio was, respectively, 1.5 and 0.71 in obese Zucker and MCD rats. While preservation damage in DCD livers is associated with the ATP/ADP recovered with OW, injury in fatty livers is linked to fatty acid constituents: livers from obese. Zucker rats, with greater content in saturated FA, might be more prone to CS injury. On the contrary, MCD livers with elevated PUFA content might be less susceptible to hypothermia.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
We compared static preservation (cold storage, CS, 4 C) with dynamic preservation (machine perfusion, MP, 20 C) followed by reperfusion using marginal livers: a model of donation after cardiac death (DCD) livers and two models of fatty livers, the methionine-choline deficient (MCD) diet model, and obese Zucker (fa/fa) rats. CS injury in DCD livers was reversed by an oxygenated washout (OW): hepatic damage, bile flow, and the ATP/ADP ratio in the OW + CS group was comparable with the ratio obtained with MP. Using fatty livers, CS preservation induced a marked release in hepatic and biliary enzymes in obese Zucker rats when compared with the MCD group. The same trend occurred for bile flow. No difference was found when comparing MP in MCD and obese Zucker rats. Fatty acid analysis demonstrated that the total saturated (SFA)/polyunsaturated fatty acid (PUFA) ratio was, respectively, 1.5 and 0.71 in obese Zucker and MCD rats. While preservation damage in DCD livers is associated with the ATP/ADP recovered with OW, injury in fatty livers is linked to fatty acid constituents: livers from obese. Zucker rats, with greater content in saturated FA, might be more prone to CS injury. On the contrary, MCD livers with elevated PUFA content might be less susceptible to hypothermia. |
Baglio SR; Lagerweij T; Pérez-Lanzón M; Ho XD; Léveillé N; Melo SA; Cleton-Jansen AM; Jordanova ES; Roncuzzi L; Greco M; van Eijndhoven MAJ; Grisendi G; Dominici M; Bonafede R; Lougheed SM; de Gruijl TD; Zini N; Cervo S; Steffan A; Canzonieri V; Martson A; Maasalu K; Köks S; Wurdinger T; Baldini N; Pegtel DM Blocking Tumor-Educated MSC Paracrine Activity Halts Osteosarcoma Progression. Journal Article In: Clinical cancer research, vol. 23, no 14, pp. 3721-3733, 2017. @article{%a1:%Y_191,
title = {Blocking Tumor-Educated MSC Paracrine Activity Halts Osteosarcoma Progression.},
author = {Baglio SR and Lagerweij T and Pérez-Lanzón M and Ho XD and Léveillé N and Melo SA and Cleton-Jansen AM and Jordanova ES and Roncuzzi L and Greco M and van Eijndhoven MAJ and Grisendi G and Dominici M and Bonafede R and Lougheed SM and de Gruijl TD and Zini N and Cervo S and Steffan A and Canzonieri V and Martson A and Maasalu K and Köks S and Wurdinger T and Baldini N and Pegtel DM},
url = {http://clincancerres.aacrjournals.org/content/23/14/3721.long},
doi = {10.1158/1078-0432.CCR-16-2726},
year = {2017},
date = {2017-02-16},
journal = {Clinical cancer research},
volume = {23},
number = {14},
pages = {3721-3733},
abstract = {Purpose: Human osteosarcoma is a genetically heterogeneous bone malignancy with poor prognosis despite the employment of aggressive chemotherapy regimens. Because druggable driver mutations have not been established, dissecting the interactions between osteosarcoma cells and supporting stroma may provide insights into novel therapeutic targets.Experimental Design: By using a bioluminescent orthotopic xenograft mouse model of osteosarcoma, we evaluated the effect of tumor extracellular vesicle (EV)-educated mesenchymal stem cells (TEMSC) on osteosarcoma progression. Characterization and functional studies were designed to assess the mechanisms underlying MSC education. Independent series of tissue specimens were analyzed to corroborate the preclinical findings, and the composition of patient serum EVs was analyzed after isolation with size-exclusion chromatography.Results: We show that EVs secreted by highly malignant osteosarcoma cells selectively incorporate a membrane-associated form of TGFbeta, which induces proinflammatory IL6 production by MSCs. TEMSCs promote tumor growth, accompanied with intratumor STAT3 activation and lung metastasis formation, which was not observed with control MSCs. Importantly, intravenous administration of the anti-IL6 receptor antibody tocilizumab abrogated the tumor-promoting effects of TEMSCs. RNA-seq analysis of human osteosarcoma tissues revealed a distinct TGFbeta-induced prometastatic gene signature. Tissue microarray immunostaining indicated active STAT3 signaling in human osteosarcoma, consistent with the observations in TEMSC-treated mice. Finally, we isolated pure populations of EVs from serum and demonstrated that circulating levels of EV-associated TGFbeta are increased in osteosarcoma patients.Conclusions: Collectively, our findings suggest that TEMSCs promote osteosarcoma progression and provide the basis for testing IL6- and TGFbeta-blocking agents as new therapeutic options for osteosarcoma patients. Clin Cancer Res; 1-13. ©2017 AACR.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Purpose: Human osteosarcoma is a genetically heterogeneous bone malignancy with poor prognosis despite the employment of aggressive chemotherapy regimens. Because druggable driver mutations have not been established, dissecting the interactions between osteosarcoma cells and supporting stroma may provide insights into novel therapeutic targets.Experimental Design: By using a bioluminescent orthotopic xenograft mouse model of osteosarcoma, we evaluated the effect of tumor extracellular vesicle (EV)-educated mesenchymal stem cells (TEMSC) on osteosarcoma progression. Characterization and functional studies were designed to assess the mechanisms underlying MSC education. Independent series of tissue specimens were analyzed to corroborate the preclinical findings, and the composition of patient serum EVs was analyzed after isolation with size-exclusion chromatography.Results: We show that EVs secreted by highly malignant osteosarcoma cells selectively incorporate a membrane-associated form of TGFbeta, which induces proinflammatory IL6 production by MSCs. TEMSCs promote tumor growth, accompanied with intratumor STAT3 activation and lung metastasis formation, which was not observed with control MSCs. Importantly, intravenous administration of the anti-IL6 receptor antibody tocilizumab abrogated the tumor-promoting effects of TEMSCs. RNA-seq analysis of human osteosarcoma tissues revealed a distinct TGFbeta-induced prometastatic gene signature. Tissue microarray immunostaining indicated active STAT3 signaling in human osteosarcoma, consistent with the observations in TEMSC-treated mice. Finally, we isolated pure populations of EVs from serum and demonstrated that circulating levels of EV-associated TGFbeta are increased in osteosarcoma patients.Conclusions: Collectively, our findings suggest that TEMSCs promote osteosarcoma progression and provide the basis for testing IL6- and TGFbeta-blocking agents as new therapeutic options for osteosarcoma patients. Clin Cancer Res; 1-13. ©2017 AACR. |
Galbiati A; Beausejour C; d'Adda di Fagagna F A novel single-cell method provides direct evidence of persistent DNA damage in senescent cells and aged mammalian tissues. Journal Article In: Aging Cell, vol. 16, no 2, pp. 422-427, 2017. @article{%a1:%Y_196,
title = {A novel single-cell method provides direct evidence of persistent DNA damage in senescent cells and aged mammalian tissues.},
author = {Galbiati A and Beausejour C and {d'Adda di Fagagna F}},
url = {http://onlinelibrary.wiley.com/doi/10.1111/acel.12573/abstract},
doi = {10.1111/acel.12573},
year = {2017},
date = {2017-02-16},
journal = {Aging Cell},
volume = {16},
number = {2},
pages = {422-427},
abstract = {The DNA damage response (DDR) arrests cell cycle progression until DNA lesions, like DNA double-strand breaks (DSBs), are repaired. The presence of DSBs in cells is usually detected by indirect techniques that rely on the accumulation of proteins at DSBs, as part of the DDR. Such detection may be biased, as some factors and their modifications may not reflect physical DNA damage. The dependency on DDR markers of DSB detection tools has left questions unanswered. In particular, it is known that senescent cells display persistent DDR foci, that we and others have proposed to be persistent DSBs, resistant to endogenous DNA repair activities. Others have proposed that these peculiar DDR foci might not be sites of damaged DNA per se but instead stable chromatin modifications, termed DNA-SCARS. Here, we developed a method, named 'DNA damage in situ ligation followed by proximity ligation assay' (DI-PLA) for the detection and imaging of DSBs in cells. DI-PLA is based on the capture of free DNA ends in fixed cells in situ, by ligation to biotinylated double-stranded DNA oligonucleotides, which are next recognized by antibiotin anti-bodies. Detection is enhanced by PLA with a partner DDR marker at the DSB. We validated DI-PLA by demonstrating its ability to detect DSBs induced by various genotoxic insults in cultured cells and tissues. Most importantly, by DI-PLA, we demonstrated that both senescent cells in culture and tissues from aged mammals retain true unrepaired DSBs associated with DDR markers. 2017 The Authors. Aging Cell published by the Anatomical Society and John Wiley & Sons Ltd.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The DNA damage response (DDR) arrests cell cycle progression until DNA lesions, like DNA double-strand breaks (DSBs), are repaired. The presence of DSBs in cells is usually detected by indirect techniques that rely on the accumulation of proteins at DSBs, as part of the DDR. Such detection may be biased, as some factors and their modifications may not reflect physical DNA damage. The dependency on DDR markers of DSB detection tools has left questions unanswered. In particular, it is known that senescent cells display persistent DDR foci, that we and others have proposed to be persistent DSBs, resistant to endogenous DNA repair activities. Others have proposed that these peculiar DDR foci might not be sites of damaged DNA per se but instead stable chromatin modifications, termed DNA-SCARS. Here, we developed a method, named 'DNA damage in situ ligation followed by proximity ligation assay' (DI-PLA) for the detection and imaging of DSBs in cells. DI-PLA is based on the capture of free DNA ends in fixed cells in situ, by ligation to biotinylated double-stranded DNA oligonucleotides, which are next recognized by antibiotin anti-bodies. Detection is enhanced by PLA with a partner DDR marker at the DSB. We validated DI-PLA by demonstrating its ability to detect DSBs induced by various genotoxic insults in cultured cells and tissues. Most importantly, by DI-PLA, we demonstrated that both senescent cells in culture and tissues from aged mammals retain true unrepaired DSBs associated with DDR markers. 2017 The Authors. Aging Cell published by the Anatomical Society and John Wiley & Sons Ltd. |
Tassini S; Sun L; Lanko K; Crespan E; Langron E; Falchi F; Kissova M; Armijos-Rivera JI; Delang L; Mirabelli C; Neyts J; Pieroni M; Cavalli A; Costantino G; Maga G; Vergani P; Leyssen P; Radi M Discovery of Multitarget Agents Active as Broad-Spectrum Antivirals and Correctors of Cystic Fibrosis Transmembrane Conductance Regulator for Associated Pulmonary Diseases. Journal Article In: Journal of medicinal chemistry, vol. 60, no 4, pp. 1400-1416, 2017. @article{%a1:%Y_199,
title = {Discovery of Multitarget Agents Active as Broad-Spectrum Antivirals and Correctors of Cystic Fibrosis Transmembrane Conductance Regulator for Associated Pulmonary Diseases.},
author = {Tassini S and Sun L and Lanko K and Crespan E and Langron E and Falchi F and Kissova M and Armijos-Rivera JI and Delang L and Mirabelli C and Neyts J and Pieroni M and Cavalli A and Costantino G and Maga G and Vergani P and Leyssen P and Radi M},
url = {http://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.6b01521},
doi = {10.1021/acs.jmedchem.6b01521},
year = {2017},
date = {2017-02-16},
journal = {Journal of medicinal chemistry},
volume = {60},
number = {4},
pages = {1400-1416},
abstract = {Enteroviruses (EVs) are among the most frequent infectious agents in humans worldwide and represent the leading cause of upper respiratory tract infections. No drugs for the treatment of EV infections are currently available. Recent studies have also linked EV infection with pulmonary exacerbations, especially in cystic fibrosis (CF) patients, and the importance of this link is probably underestimated. The aim of this work was to develop a new class of multitarget agents active both as broad-spectrum antivirals and as correctors of the F508del-cystic fibrosis transmembrane conductance regulator (CFTR) folding defect responsible for >90% of CF cases. We report herein the discovery of the first small molecules able to simultaneously act as correctors of the F508del-CFTR folding defect and as broad-spectrum antivirals against a panel of EVs representative of all major species.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Enteroviruses (EVs) are among the most frequent infectious agents in humans worldwide and represent the leading cause of upper respiratory tract infections. No drugs for the treatment of EV infections are currently available. Recent studies have also linked EV infection with pulmonary exacerbations, especially in cystic fibrosis (CF) patients, and the importance of this link is probably underestimated. The aim of this work was to develop a new class of multitarget agents active both as broad-spectrum antivirals and as correctors of the F508del-cystic fibrosis transmembrane conductance regulator (CFTR) folding defect responsible for >90% of CF cases. We report herein the discovery of the first small molecules able to simultaneously act as correctors of the F508del-CFTR folding defect and as broad-spectrum antivirals against a panel of EVs representative of all major species. |
Abou Khouzam R; Molinari C; Salvi S; Marabelli M; Molinaro V; Orioli D; Saragoni L; Morgagni P; Calistri D; Ranzani GN Digital PCR identifies changes in CDH1 (E-cadherin) transcription pattern in intestinal-type gastric cancer. Journal Article In: Oncotarget, vol. 8, no 12, pp. 18811-18820, 2017. @article{%a1:%Y_203,
title = {Digital PCR identifies changes in CDH1 (E-cadherin) transcription pattern in intestinal-type gastric cancer.},
author = {{Abou Khouzam R} and Molinari C and Salvi S and Marabelli M and Molinaro V and Orioli D and Saragoni L and Morgagni P and Calistri D and Ranzani GN},
url = {http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path[]=13401&pubmed-linkout=1},
doi = {10.18632/oncotarget.13401},
year = {2017},
date = {2017-02-16},
journal = {Oncotarget},
volume = {8},
number = {12},
pages = {18811-18820},
abstract = {E-cadherin is a cell-cell adhesion protein encoded by CDH1 tumor-suppressor gene. CDH1 inactivating mutations, leading to loss of protein expression, are common in gastric cancer of the diffuse histotype, while alternative mechanisms modulating E-cadherin expression characterize the more common intestinal histotype. These mechanisms are still poorly understood. CDH1 intron 2 has recently emerged as a cis-modulator of E-cadherin expression, encoding non-canonical transcripts. One in particular, CDH1a, proved to be expressed in gastric cancer cell lines, while being absent in the normal stomach. For the first time, we evaluated by digital PCR the expression of CDH1 and CDH1a transcripts in cancer and normal tissue samples from 32 patients with intestinal-type gastric cancer. We found a significant decrease in CDH1 expression in tumors compared to normal counterparts (P = 0.001), which was especially evident in 76% of cases. CDH1a was detected at extremely low levels in 47% of tumors, but not in normal mucosa. A trend was observed of having less CDH1 in tumors expressing CDH1atranscript. The majority of tumors with both a decrease in CDH1 and presence of CDH1a also showed a decrease in miR-101 expression levels. On the whole, the decrease of CDH1 transcript, corresponding to the canonical protein, and the presence of CDH1a, corresponding to an alternative isoform, are likely to perturb E-cadherin-mediated signaling and cell-cell adhesion, thus contributing to intestinal-type gastric carcinogenesis.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
E-cadherin is a cell-cell adhesion protein encoded by CDH1 tumor-suppressor gene. CDH1 inactivating mutations, leading to loss of protein expression, are common in gastric cancer of the diffuse histotype, while alternative mechanisms modulating E-cadherin expression characterize the more common intestinal histotype. These mechanisms are still poorly understood. CDH1 intron 2 has recently emerged as a cis-modulator of E-cadherin expression, encoding non-canonical transcripts. One in particular, CDH1a, proved to be expressed in gastric cancer cell lines, while being absent in the normal stomach. For the first time, we evaluated by digital PCR the expression of CDH1 and CDH1a transcripts in cancer and normal tissue samples from 32 patients with intestinal-type gastric cancer. We found a significant decrease in CDH1 expression in tumors compared to normal counterparts (P = 0.001), which was especially evident in 76% of cases. CDH1a was detected at extremely low levels in 47% of tumors, but not in normal mucosa. A trend was observed of having less CDH1 in tumors expressing CDH1atranscript. The majority of tumors with both a decrease in CDH1 and presence of CDH1a also showed a decrease in miR-101 expression levels. On the whole, the decrease of CDH1 transcript, corresponding to the canonical protein, and the presence of CDH1a, corresponding to an alternative isoform, are likely to perturb E-cadherin-mediated signaling and cell-cell adhesion, thus contributing to intestinal-type gastric carcinogenesis. |
Schiavone M; Zulian A; Menazza S; Petronilli V; Argenton F; Merlini L; Sabatelli P; Bernardi P Alisporivir rescues defective mitochondrial respiration in Duchenne muscular dystrophy. Journal Article In: Pharmacological research, vol. 125, no Pt, pp. 122-131, 2017. @article{%a1:%Y_206,
title = {Alisporivir rescues defective mitochondrial respiration in Duchenne muscular dystrophy.},
author = {Schiavone M and Zulian A and Menazza S and Petronilli V and Argenton F and Merlini L and Sabatelli P and Bernardi P},
url = {http://www.sciencedirect.com/science/article/pii/S1043661817309386?via%3Dihub},
doi = {10.1016/j.phrs.2017.09.001},
year = {2017},
date = {2017-02-16},
journal = {Pharmacological research},
volume = {125},
number = {Pt},
pages = {122-131},
abstract = {Duchenne muscular dystrophy (DMD) is a severe muscle disease of known etiology without effective, or generally applicable therapy. Mitochondria are affected by the disease in animal models but whether mitochondrial dysfunction is part of the pathogenesis in patients remains unclear. We show that primary cultures obtained from muscle biopsies of DMD patients display a decrease of the respiratory reserve, a consequence of inappropriate opening of the permeability transition pore (PTP). Treatment with the cyclophilin inhibitor alisporivir - a cyclosporin A derivative that desensitizes the PTP but does not inhibit calcineurin - largely restored the maximal respiratory capacity without affecting basal oxygen consumption in cells from patients, thus reinstating a normal respiratory reserve. Treatment with alisporivir, but not with cyclosporin A, led to a substantial recovery of respiratory function matching improved muscle ultrastructure and survival of sapje zebrafish, a severe model of DMD where muscle defects are close to those of DMD patients. Alisporivir was generally well tolerated in HCV patients and could be used for the treatment of DMD.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Duchenne muscular dystrophy (DMD) is a severe muscle disease of known etiology without effective, or generally applicable therapy. Mitochondria are affected by the disease in animal models but whether mitochondrial dysfunction is part of the pathogenesis in patients remains unclear. We show that primary cultures obtained from muscle biopsies of DMD patients display a decrease of the respiratory reserve, a consequence of inappropriate opening of the permeability transition pore (PTP). Treatment with the cyclophilin inhibitor alisporivir - a cyclosporin A derivative that desensitizes the PTP but does not inhibit calcineurin - largely restored the maximal respiratory capacity without affecting basal oxygen consumption in cells from patients, thus reinstating a normal respiratory reserve. Treatment with alisporivir, but not with cyclosporin A, led to a substantial recovery of respiratory function matching improved muscle ultrastructure and survival of sapje zebrafish, a severe model of DMD where muscle defects are close to those of DMD patients. Alisporivir was generally well tolerated in HCV patients and could be used for the treatment of DMD. |
