Cipolla L; Maffia A; Bertoletti F; Sabbioneda S The Regulation of DNA Damage Tolerance by Ubiquitin and Ubiquitin-Like Modifiers. Journal Article In: Frontiers in Genetics, vol. 7, pp. 105, 2016. @article{%a1:%Y_262,
title = {The Regulation of DNA Damage Tolerance by Ubiquitin and Ubiquitin-Like Modifiers.},
author = {Cipolla L and Maffia A and Bertoletti F and Sabbioneda S},
url = {http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4904029/},
doi = {10.3389/fgene.2016.00105},
year = {2016},
date = {2016-03-11},
journal = {Frontiers in Genetics},
volume = {7},
pages = {105},
abstract = {DNA replication is an extremely complex process that needs to be executed in a highly accurate manner in order to propagate the genome. This task requires the coordination of a number of enzymatic activities and it is fragile and prone to arrest after DNA damage. DNA damage tolerance provides a last line of defense that allows completion of DNA replication in the presence of an unrepaired template. One of such mechanisms is called post-replication repair (PRR) and it is used by the cells to bypass highly distorted templates caused by damaged bases. PRR is extremely important for the cellular life and performs the bypass of the damage both in an error-free and in an error-prone manner. In light of these two possible outcomes, PRR needs to be tightly controlled in order to prevent the accumulation of mutations leading ultimately to genome instability. Post-translational modifications of PRR proteins provide the framework for this regulation with ubiquitylation and SUMOylation playing a pivotal role in choosing which pathway to activate, thus controlling the different outcomes of damage bypass. The proliferating cell nuclear antigen (PCNA), the DNA clamp for replicative polymerases, plays a central role in the regulation of damage tolerance and its modification by ubiquitin, and SUMO controls both the error-free and error-prone branches of PRR. Furthermore, a significant number of polymerases are involved in the bypass of DNA damage possess domains that can bind post-translational modifications and they are themselves target for ubiquitylation. In this review, we will focus on how ubiquitin and ubiquitin-like modifications can regulate the DNA damage tolerance systems and how they control the recruitment of different proteins to the replication fork.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
DNA replication is an extremely complex process that needs to be executed in a highly accurate manner in order to propagate the genome. This task requires the coordination of a number of enzymatic activities and it is fragile and prone to arrest after DNA damage. DNA damage tolerance provides a last line of defense that allows completion of DNA replication in the presence of an unrepaired template. One of such mechanisms is called post-replication repair (PRR) and it is used by the cells to bypass highly distorted templates caused by damaged bases. PRR is extremely important for the cellular life and performs the bypass of the damage both in an error-free and in an error-prone manner. In light of these two possible outcomes, PRR needs to be tightly controlled in order to prevent the accumulation of mutations leading ultimately to genome instability. Post-translational modifications of PRR proteins provide the framework for this regulation with ubiquitylation and SUMOylation playing a pivotal role in choosing which pathway to activate, thus controlling the different outcomes of damage bypass. The proliferating cell nuclear antigen (PCNA), the DNA clamp for replicative polymerases, plays a central role in the regulation of damage tolerance and its modification by ubiquitin, and SUMO controls both the error-free and error-prone branches of PRR. Furthermore, a significant number of polymerases are involved in the bypass of DNA damage possess domains that can bind post-translational modifications and they are themselves target for ubiquitylation. In this review, we will focus on how ubiquitin and ubiquitin-like modifications can regulate the DNA damage tolerance systems and how they control the recruitment of different proteins to the replication fork. |
Sardone F; Traina F; Bondi A; Merlini L; Santi S; Maraldi NM; Faldini C; Sabatelli P Tendon Extracellular Matrix Alterations in Ullrich Congenital Muscular Dystrophy. Journal Article In: Frontiers in Aging Neuroscience, vol. 8, pp. 131, 2016. @article{%a1:%Y_308,
title = {Tendon Extracellular Matrix Alterations in Ullrich Congenital Muscular Dystrophy.},
author = {Sardone F and Traina F and Bondi A and Merlini L and Santi S and Maraldi NM and Faldini C and Sabatelli P},
url = {http://journal.frontiersin.org/article/10.3389/fnagi.2016.00131/full},
doi = {10.3389/fnagi.2016.00131},
year = {2016},
date = {2016-03-10},
journal = {Frontiers in Aging Neuroscience},
volume = {8},
pages = {131},
abstract = {Collagen VI (COLVI) is a non-fibrillar collagen expressed in skeletal muscle and most connective tissues. Mutations in COLVI genes cause two major clinical forms, Bethlem myopathy and Ullrich congenital muscular dystrophy (UCMD). In addition to congenital muscle weakness, patients affected by COLVI myopathies show axial and proximal joint contractures and distal joint hypermobility, which suggest the involvement of the tendon function. We examined a peroneal tendon biopsy and tenocyte culture of a 15-year-old patient affected by UCMD with compound heterozygous COL6A2 mutations. In patient's tendon biopsy, we found striking morphological alterations of tendon fibrils, consisting in irregular profiles and reduced mean diameter. The organization of the pericellular matrix of tenocytes, the primary site of collagen fibril assembly, was severely affected, as determined by immunoelectron microscopy, which showed an abnormal accumulation of COLVI and altered distribution of collagen I (COLI) and fibronectin (FBN). In patient's tenocyte culture, COLVI web formation and cell surface association were severely impaired; large aggregates of COLVI, which matched with COLI labeling, were frequently detected in the extracellular matrix. In addition, metalloproteinase MMP-2, an extracellular matrix-regulating enzyme, was increased in the conditioned medium of patient's tenocytes, as determined by gelatin zymography and western blot. Altogether, these data indicate that COLVI deficiency may influence the organization of UCMD tendon matrix, resulting in dysfunctional fibrillogenesis. The alterations of tendon matrix may contribute to the complex pathogenesis of COLVI related myopathies.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Collagen VI (COLVI) is a non-fibrillar collagen expressed in skeletal muscle and most connective tissues. Mutations in COLVI genes cause two major clinical forms, Bethlem myopathy and Ullrich congenital muscular dystrophy (UCMD). In addition to congenital muscle weakness, patients affected by COLVI myopathies show axial and proximal joint contractures and distal joint hypermobility, which suggest the involvement of the tendon function. We examined a peroneal tendon biopsy and tenocyte culture of a 15-year-old patient affected by UCMD with compound heterozygous COL6A2 mutations. In patient's tendon biopsy, we found striking morphological alterations of tendon fibrils, consisting in irregular profiles and reduced mean diameter. The organization of the pericellular matrix of tenocytes, the primary site of collagen fibril assembly, was severely affected, as determined by immunoelectron microscopy, which showed an abnormal accumulation of COLVI and altered distribution of collagen I (COLI) and fibronectin (FBN). In patient's tenocyte culture, COLVI web formation and cell surface association were severely impaired; large aggregates of COLVI, which matched with COLI labeling, were frequently detected in the extracellular matrix. In addition, metalloproteinase MMP-2, an extracellular matrix-regulating enzyme, was increased in the conditioned medium of patient's tenocytes, as determined by gelatin zymography and western blot. Altogether, these data indicate that COLVI deficiency may influence the organization of UCMD tendon matrix, resulting in dysfunctional fibrillogenesis. The alterations of tendon matrix may contribute to the complex pathogenesis of COLVI related myopathies. |
Kissova M; Maga G; Crespan E The human tyrosine kinase Kit and its gatekeeper mutant T670I, show different kinetic properties: Implications for drug design. Journal Article In: Bioorganic & Medicinal Chemistry, vol. 24, no 19, pp. 4555-4532, 2016. @article{%a1:%Y_321,
title = {The human tyrosine kinase Kit and its gatekeeper mutant T670I, show different kinetic properties: Implications for drug design.},
author = {Kissova M and Maga G and Crespan E},
url = {http://www.sciencedirect.com/science/article/pii/S0968089616305806},
doi = {10.1016/j.bmc.2016.07.059},
year = {2016},
date = {2016-03-10},
journal = {Bioorganic & Medicinal Chemistry},
volume = {24},
number = {19},
pages = {4555-4532},
abstract = {The tyrosine kinase Kit, a receptor for Stem Cell Factor, is involved, among others, in processes associated to cell survival, proliferation and migration. Upon physiological conditions, the activity of Kit is tightly regulated. However, primary mutations that lead to its constitutive activation are the causal oncogenic driver of gastrointestinal stromal tumours (GISTs). GISTs are known to be refractory to conventional therapies but the introduction of Imatinib, a selective inhibitor of tyrosine kinases Abl and Kit, significantly ameliorated the treatment options of GISTs patients. However, the acquisition of secondary mutations renders Kit resistant towards all available drugs. Mutation involving gatekeeper residues (such as V654a and T670I) influence both the structure and the catalytic activity of the enzyme. Therefore, detailed knowledge of the enzymatic properties of the mutant forms, in comparison with the wild type enzyme, is an important pre-requisite for the rational development of specific inhibitors. In this paper we report a thorough kinetic analysis of the reaction catalyzed by the Kit kinase and its gatekeeper mutated form T670I. Our results revealed the different mechanisms of action of these two enzymes and may open a new avenue for the future design of specific Kit inhibitors.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The tyrosine kinase Kit, a receptor for Stem Cell Factor, is involved, among others, in processes associated to cell survival, proliferation and migration. Upon physiological conditions, the activity of Kit is tightly regulated. However, primary mutations that lead to its constitutive activation are the causal oncogenic driver of gastrointestinal stromal tumours (GISTs). GISTs are known to be refractory to conventional therapies but the introduction of Imatinib, a selective inhibitor of tyrosine kinases Abl and Kit, significantly ameliorated the treatment options of GISTs patients. However, the acquisition of secondary mutations renders Kit resistant towards all available drugs. Mutation involving gatekeeper residues (such as V654a and T670I) influence both the structure and the catalytic activity of the enzyme. Therefore, detailed knowledge of the enzymatic properties of the mutant forms, in comparison with the wild type enzyme, is an important pre-requisite for the rational development of specific inhibitors. In this paper we report a thorough kinetic analysis of the reaction catalyzed by the Kit kinase and its gatekeeper mutated form T670I. Our results revealed the different mechanisms of action of these two enzymes and may open a new avenue for the future design of specific Kit inhibitors. |
Giannaccare G; Blalock WL; Fresina M; Vagge A; Versura P Intolerant contact lens wearers exhibit ocular surface impairment despite 3 months wear discontinuation. Journal Article In: Graefe's Archive for Clinical and Experimental Ophthalmology , vol. 254, no 9, pp. 1825-1831, 2016. @article{%a1:%Y_283,
title = {Intolerant contact lens wearers exhibit ocular surface impairment despite 3 months wear discontinuation.},
author = {Giannaccare G and Blalock WL and Fresina M and Vagge A and Versura P},
url = {http://link.springer.com/article/10.1007%2Fs00417-016-3400-4},
doi = {10.1007/s00417-016-3400-4},
year = {2016},
date = {2016-03-09},
journal = {Graefe's Archive for Clinical and Experimental Ophthalmology },
volume = {254},
number = {9},
pages = {1825-1831},
abstract = {Purpose: To evaluate ocular surface (OS) parameters recovery in intolerant contact lens (CL) wearers after a period of discontinuation. Methods: This is a retrospective analysis of data from 87 intolerant CL wearers who had discontinued their use for an average period of 12 weeks because of associated discomfort and failure to successfully refit. Data were collected from clinical charts. Data from 50 matched healthy volunteers served as controls. Clinical tests included subjective discomfort symptoms questionnaire (Ocular Surface Disease Index, OSDI), Schirmer test, break-up time (BUT), corneal esthesiometry and corneo-conjunctival staining. Laboratory tests included scraping and imprint cytology. Tear protein analysis included dosage of total tear protein (TP), lysozyme-C (LYS-C), lactoferrin (LACTO), zinc-α2-glycoprotein (ZAG-2), IgA heavy chain bands (Ig-A), and serum albumin (ALB). Data were correlated to wear parameters. Results: All values were significantly worse in intolerant CL wearers group (p always <0.001). In particular, lower values compared to controls were found for BUT, corneal esthesiometry, goblet cell density, LYS-C, LACTO, ZAG-2, and TP. On the contrary, higher values compared to controls were found for OSDI, staining, imprint cytology, scraping cytology, ALB, IgA-heavy chain. The IgA/LYS-C ratio calculated as an index of the increased activity of the IgA-producing cell was found significantly higher in the intolerant group and in correlation with discomfort symptoms. Conclusions: Ocular surface parameters were altered in intolerant CL wearers, even after a prolonged discontinuation period. Our data suggest that OS recovery necessary to successfully refit lenses may need a discontinuation time longer than 3 months.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Purpose: To evaluate ocular surface (OS) parameters recovery in intolerant contact lens (CL) wearers after a period of discontinuation. Methods: This is a retrospective analysis of data from 87 intolerant CL wearers who had discontinued their use for an average period of 12 weeks because of associated discomfort and failure to successfully refit. Data were collected from clinical charts. Data from 50 matched healthy volunteers served as controls. Clinical tests included subjective discomfort symptoms questionnaire (Ocular Surface Disease Index, OSDI), Schirmer test, break-up time (BUT), corneal esthesiometry and corneo-conjunctival staining. Laboratory tests included scraping and imprint cytology. Tear protein analysis included dosage of total tear protein (TP), lysozyme-C (LYS-C), lactoferrin (LACTO), zinc-α2-glycoprotein (ZAG-2), IgA heavy chain bands (Ig-A), and serum albumin (ALB). Data were correlated to wear parameters. Results: All values were significantly worse in intolerant CL wearers group (p always <0.001). In particular, lower values compared to controls were found for BUT, corneal esthesiometry, goblet cell density, LYS-C, LACTO, ZAG-2, and TP. On the contrary, higher values compared to controls were found for OSDI, staining, imprint cytology, scraping cytology, ALB, IgA-heavy chain. The IgA/LYS-C ratio calculated as an index of the increased activity of the IgA-producing cell was found significantly higher in the intolerant group and in correlation with discomfort symptoms. Conclusions: Ocular surface parameters were altered in intolerant CL wearers, even after a prolonged discontinuation period. Our data suggest that OS recovery necessary to successfully refit lenses may need a discontinuation time longer than 3 months. |
Aredia F; Scovassi AI Manipulation of Intracellular pH in Cancer Cells by NHE1 Inhibitors. Journal Article In: Protein and peptide letters, vol. 23, no 12, pp. 1123-1129, 2016. @article{%a1:%Y_246,
title = {Manipulation of Intracellular pH in Cancer Cells by NHE1 Inhibitors.},
author = {Aredia F and Scovassi AI},
url = {http://www.eurekaselect.com/146253/article},
doi = {10.2174/0929866523666161013125536 },
year = {2016},
date = {2016-03-08},
journal = {Protein and peptide letters},
volume = {23},
number = {12},
pages = {1123-1129},
abstract = {Cancer cells are characterized by a peculiar pH condition, being the extracellular compartment acidic and the intracellular one neutral or basic, i.e. the opposite of what happens in normal cells. The reversal of the pH contributes to cancer cell proliferation and drug resistance. Among the different enzymes regulating pH gradient, proton transporters Na+/H+ exchangers (NHEs) are considered as suitable targets for drugs that ultimately counteract cancer cell survival. This review will describe the properties of NHEs, focusing on the prototype NHE1 and on the effect of its inhibition on cancer cell metabolism.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Cancer cells are characterized by a peculiar pH condition, being the extracellular compartment acidic and the intracellular one neutral or basic, i.e. the opposite of what happens in normal cells. The reversal of the pH contributes to cancer cell proliferation and drug resistance. Among the different enzymes regulating pH gradient, proton transporters Na+/H+ exchangers (NHEs) are considered as suitable targets for drugs that ultimately counteract cancer cell survival. This review will describe the properties of NHEs, focusing on the prototype NHE1 and on the effect of its inhibition on cancer cell metabolism. |
Bellotti C; Capanni C; Lattanzi G; Donati D; Lucarelli E; Duchi S Detection of mesenchymal stem cells senescence by prelamin A accumulation at the nuclear level. Journal Article In: Springerplus, vol. 5, no 1, pp. 1427, 2016. @article{%a1:%Y_253,
title = {Detection of mesenchymal stem cells senescence by prelamin A accumulation at the nuclear level.},
author = {Bellotti C and Capanni C and Lattanzi G and Donati D and Lucarelli E and Duchi S},
url = {http://springerplus.springeropen.com/articles/10.1186/s40064-016-3091-7},
doi = {10.1186/s40064-016-3091-7},
year = {2016},
date = {2016-03-08},
urldate = {2016-03-08},
journal = {Springerplus},
volume = {5},
number = {1},
pages = {1427},
abstract = {Human mesenchymal stem cells (MSC), during in vitro expansion, undergo a progressive loss of proliferative potential that leads to the senescent state, associated with a reduction of their "medicinal" properties. This may hampers their efficacy in the treatment of injured tissues. Quality controls on MSC-based cell therapy products should include an assessment of the senescent state. However, a reliable and specific marker is still missing. From studies on lamin-associated disorders, has emerged the correlation between defective lamin A maturation and cellular senescence. FINDINGS: Primary cultured hMSC lines (n = 3), were analyzed by immunostaining at different life-span stages for the accumulation of prelamin A, along with other markers of cellular senescence. During culture, cells at the last stage of their life span displayed evident signs of senescence consistent with the positivity of SA-β-gal staining. We also observed a significant increase of prelamin A positive cells. Furthermore, we verified that the cells marked by prelamin A were also positive for p21(Waf1) while negative for Ki67. CONCLUSIONS: Overall data support that the detection of prelamin A identifies senescent MSC, providing an easy and reliable tool to be use alone or in combination with known senescence markers to screen MSC before their use in clinical applications.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Human mesenchymal stem cells (MSC), during in vitro expansion, undergo a progressive loss of proliferative potential that leads to the senescent state, associated with a reduction of their "medicinal" properties. This may hampers their efficacy in the treatment of injured tissues. Quality controls on MSC-based cell therapy products should include an assessment of the senescent state. However, a reliable and specific marker is still missing. From studies on lamin-associated disorders, has emerged the correlation between defective lamin A maturation and cellular senescence. FINDINGS: Primary cultured hMSC lines (n = 3), were analyzed by immunostaining at different life-span stages for the accumulation of prelamin A, along with other markers of cellular senescence. During culture, cells at the last stage of their life span displayed evident signs of senescence consistent with the positivity of SA-β-gal staining. We also observed a significant increase of prelamin A positive cells. Furthermore, we verified that the cells marked by prelamin A were also positive for p21(Waf1) while negative for Ki67. CONCLUSIONS: Overall data support that the detection of prelamin A identifies senescent MSC, providing an easy and reliable tool to be use alone or in combination with known senescence markers to screen MSC before their use in clinical applications. |
Loi M; Cenni V; Duchi S; Squarzoni S; Lopez-Otin C; Foisner R; Lattanzi G; Capanni C Barrier-to-Autointegration Factor (BAF) involvement in prelamin A-related chromatin organization changes. Journal Article In: Oncotarget, vol. 7, no 13, pp. 15662-15677, 2016. @article{%a1:%Y_293,
title = {Barrier-to-Autointegration Factor (BAF) involvement in prelamin A-related chromatin organization changes.},
author = {Loi M and Cenni V and Duchi S and Squarzoni S and {Lopez-Otin C} and Foisner R and Lattanzi G and Capanni C},
url = {http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path[]=6697&pubmed-linkout=1},
doi = {10.18632/oncotarget.6697},
year = {2016},
date = {2016-03-08},
urldate = {2016-03-08},
journal = {Oncotarget},
volume = {7},
number = {13},
pages = {15662-15677},
abstract = {Chromatin disorganization is one of the major alterations linked to prelamin A processing impairment. In this study we demonstrate that BAF is necessary to modulate prelamin A effects on chromatin structure. We show that when prelamin A and BAF cannot properly interact no prelamin A-dependent effects on chromatin occur; similar to what is observed in human Nestor Guillermo Progeria Syndrome cells harboring a BAF mutation, in HEK293 cells expressing a BAF mutant unable to bind prelamin A, or in siRNA mediated BAF-depleted HEK293 cells expressing prelamin A. BAF is necessary to induce histone trimethyl-H3K9 as well as HP1-alpha and LAP2-alpha nuclear relocalization in response to prelamin A accumulation. These findings are enforced by electron microscopy evaluations showing how the prelamin A-BAF interaction governs overall chromatin organization. Finally, we demonstrate that the LAP2-alpha nuclear localization defect observed in HGPS cells involves the progerin-BAF interaction, thus establishing a functional link between BAF and prelamin A pathological forms.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Chromatin disorganization is one of the major alterations linked to prelamin A processing impairment. In this study we demonstrate that BAF is necessary to modulate prelamin A effects on chromatin structure. We show that when prelamin A and BAF cannot properly interact no prelamin A-dependent effects on chromatin occur; similar to what is observed in human Nestor Guillermo Progeria Syndrome cells harboring a BAF mutation, in HEK293 cells expressing a BAF mutant unable to bind prelamin A, or in siRNA mediated BAF-depleted HEK293 cells expressing prelamin A. BAF is necessary to induce histone trimethyl-H3K9 as well as HP1-alpha and LAP2-alpha nuclear relocalization in response to prelamin A accumulation. These findings are enforced by electron microscopy evaluations showing how the prelamin A-BAF interaction governs overall chromatin organization. Finally, we demonstrate that the LAP2-alpha nuclear localization defect observed in HGPS cells involves the progerin-BAF interaction, thus establishing a functional link between BAF and prelamin A pathological forms. |
Ramazzotti G; Bavelloni A; Blalock WL; Piazzi M; Cocco L; Faenza I BMP-2 Induced Expression of PLCbeta1 That Is a Positive Regulator of Osteoblast Differentiation. Journal Article In: Journal of Cellular Physiology, vol. 231, no 3, pp. 623-629, 2016. @article{%a1:%Y_305,
title = {BMP-2 Induced Expression of PLCbeta1 That Is a Positive Regulator of Osteoblast Differentiation.},
author = {Ramazzotti G and Bavelloni A and Blalock WL and Piazzi M and Cocco L and Faenza I},
url = {http://onlinelibrary.wiley.com/doi/10.1002/jcp.25107/abstract;jsessionid=9619766386605F1FBD94DD07A67E9B42.f04t02},
year = {2016},
date = {2016-03-07},
journal = {Journal of Cellular Physiology},
volume = {231},
number = {3},
pages = {623-629},
abstract = {"Bone morphogenetic protein 2 (BMP-2) is a critical growth factor that directs osteoblast differentiation and bone formation. Phosphoinositide-phospholipase Cbeta 1 (PLCbeta1) plays a crucial role in the initiation of the genetic program responsible for muscle differentiation. Differentiation of C2C12 mouse myoblasts in response to insulin stimulation is characterized by a marked increase in nuclear PLCbeta1. Here, the function of PLCbeta1 in the osteogenic differentiation was investigated. Briefly, in C2C12 cells treated with BMP-2 we assist to a remarkable increase in PLCbeta1 protein and mRNA expression. The data regarding the influence on differentiation demonstrated that PLCbeta1 promotes osteogenic differentiation by up-regulating alkaline phosphatase (ALP). Moreover, PLCbeta1 is present in the nuclear compartment of these cells and overexpression of a cytosolic-PLCbeta1mutant (cyt-PLCbeta1), which lacks a nuclear localization sequence, prevented the differentiation of C2C12 cells into osteocytes. Recent evidence indicates that miRNAs act as important post transcriptional regulators in a large number of processes, including osteoblast differentiation. Since miR-214 is a regulator of Osterix (Osx) which is an osteoblast-specific transcription factor that is needful for osteoblast differentiation and bone formation, we further investigated whether PLCbeta1 could be a potential target of miR-214 in the control of osteogenic differentiation by gain- and loss- of function experiment. The results indicated that inhibition of miR-214 in C2C12 cells significantly enhances the protein level of PLCbeta1 and promotes C2C12 BMP-2-induced osteogenesis by targeting PLCbeta1. This article is protected by copyright. All rights reserved.This article is protected by copyright. All rights reserved.
"},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
"Bone morphogenetic protein 2 (BMP-2) is a critical growth factor that directs osteoblast differentiation and bone formation. Phosphoinositide-phospholipase Cbeta 1 (PLCbeta1) plays a crucial role in the initiation of the genetic program responsible for muscle differentiation. Differentiation of C2C12 mouse myoblasts in response to insulin stimulation is characterized by a marked increase in nuclear PLCbeta1. Here, the function of PLCbeta1 in the osteogenic differentiation was investigated. Briefly, in C2C12 cells treated with BMP-2 we assist to a remarkable increase in PLCbeta1 protein and mRNA expression. The data regarding the influence on differentiation demonstrated that PLCbeta1 promotes osteogenic differentiation by up-regulating alkaline phosphatase (ALP). Moreover, PLCbeta1 is present in the nuclear compartment of these cells and overexpression of a cytosolic-PLCbeta1mutant (cyt-PLCbeta1), which lacks a nuclear localization sequence, prevented the differentiation of C2C12 cells into osteocytes. Recent evidence indicates that miRNAs act as important post transcriptional regulators in a large number of processes, including osteoblast differentiation. Since miR-214 is a regulator of Osterix (Osx) which is an osteoblast-specific transcription factor that is needful for osteoblast differentiation and bone formation, we further investigated whether PLCbeta1 could be a potential target of miR-214 in the control of osteogenic differentiation by gain- and loss- of function experiment. The results indicated that inhibition of miR-214 in C2C12 cells significantly enhances the protein level of PLCbeta1 and promotes C2C12 BMP-2-induced osteogenesis by targeting PLCbeta1. This article is protected by copyright. All rights reserved.This article is protected by copyright. All rights reserved.
" |
Manescu A; Giuliani A; Mohammadi S; Tromba G; Mazzoni S; Diomede F; Zini N; Piattelli A; Trubiani O Osteogenic potential of dualblocks cultured with human periodontal ligament stem cells: in vitro and synchrotron microtomography study. Journal Article In: Journal of Periodontal Research, vol. 51, no 1, pp. 112-124, 2016. @article{%a1:%Y_296,
title = {Osteogenic potential of dualblocks cultured with human periodontal ligament stem cells: in vitro and synchrotron microtomography study.},
author = {Manescu A and Giuliani A and Mohammadi S and Tromba G and Mazzoni S and Diomede F and Zini N and Piattelli A and Trubiani O},
url = {http://onlinelibrary.wiley.com/doi/10.1111/jre.12289/abstract},
doi = {10.1111/jre.12289},
year = {2016},
date = {2016-02-29},
journal = {Journal of Periodontal Research},
volume = {51},
number = {1},
pages = {112-124},
abstract = {"BACKGROUND AND OBJECTIVE: In the present study, the early stages of in vitro bone formation in collagenated porcine scaffolds cultured with human periodontal ligament cells were investigated. The comparison between the osteogenic potential of this structure in basal and differentiating culture media was explored to predict the mechanism of its biological behavior as graft in human defect. Results were validated by synchrotron radiation X-Ray phase contrast computed microtomography (micro-CT). As the periodontal disease plays a key role in systemic and oral diseases, it is crucial to find advanced therapeutic clinical interventions to repair periodontal defects. This has been recently explored using cells and tissues developed in vitro that should ideally be immunologically, functionally, structurally and mechanically identical to the native tissue. MATERIAL AND METHODS:
In vitro cultures of human periodontal ligament cells, easily obtained by scraping of alveolar crestal and horizontal fibers of the periodontal ligament, were seeded on to collagenated porcine blocks constituted by natural cancellous and cortical bone. 3D images were obtained by synchrotron radiation micro-CT and processed with a phase-retrieval algorithm based on the transport of intensity equation. RESULTS: Starting from the second week of culture, newly formed mineralized bone was detected in all the scaffolds, both in basal and differentiating media. Bone mineralization was proved to occur preferentially in the trabecular portion and in differentiating media. CONCLUSION: The chosen method, supported by phase contrast micro-CT analysis, successfully and quantitatively monitored the early stages of bone formation and the rate of the bioscaffold resorption in basal and differentiating culture media. 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd."},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
"BACKGROUND AND OBJECTIVE: In the present study, the early stages of in vitro bone formation in collagenated porcine scaffolds cultured with human periodontal ligament cells were investigated. The comparison between the osteogenic potential of this structure in basal and differentiating culture media was explored to predict the mechanism of its biological behavior as graft in human defect. Results were validated by synchrotron radiation X-Ray phase contrast computed microtomography (micro-CT). As the periodontal disease plays a key role in systemic and oral diseases, it is crucial to find advanced therapeutic clinical interventions to repair periodontal defects. This has been recently explored using cells and tissues developed in vitro that should ideally be immunologically, functionally, structurally and mechanically identical to the native tissue. MATERIAL AND METHODS:
In vitro cultures of human periodontal ligament cells, easily obtained by scraping of alveolar crestal and horizontal fibers of the periodontal ligament, were seeded on to collagenated porcine blocks constituted by natural cancellous and cortical bone. 3D images were obtained by synchrotron radiation micro-CT and processed with a phase-retrieval algorithm based on the transport of intensity equation. RESULTS: Starting from the second week of culture, newly formed mineralized bone was detected in all the scaffolds, both in basal and differentiating media. Bone mineralization was proved to occur preferentially in the trabecular portion and in differentiating media. CONCLUSION: The chosen method, supported by phase contrast micro-CT analysis, successfully and quantitatively monitored the early stages of bone formation and the rate of the bioscaffold resorption in basal and differentiating culture media. 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd." |
Okbay A; Beauchamp JP; Fontana MA; Lee JJ; Pers TH; Rietveld CA; Turley P; Chen GB; Emilsson V; Meddens SF; Oskarsson S; Pickrell JK; Thom K; Timshel P; et al; Biino G; et al; Benjamin DJ Genome-wide association study identifies 74 loci associated with educational attainment. Journal Article In: Nature, vol. 533, no 7604, pp. 539-542, 2016. @article{%a1:%Y_302,
title = {Genome-wide association study identifies 74 loci associated with educational attainment.},
author = {Okbay A and Beauchamp JP and Fontana MA and Lee JJ and Pers TH and Rietveld CA and Turley P and Chen GB and Emilsson V and Meddens SF and Oskarsson S and Pickrell JK and Thom K and Timshel P and {et al} and Biino G and {et al} and Benjamin DJ},
url = {http://www.nature.com/nature/journal/v533/n7604/full/nature17671.html},
doi = {10.1038/nature17671},
year = {2016},
date = {2016-02-29},
journal = {Nature},
volume = {533},
number = {7604},
pages = {539-542},
abstract = {Educational attainment is strongly influenced by social and other environmental factors, but genetic factors are estimated to account for at least 20% of the variation across individuals. Here we report the results of a genome-wide association study (GWAS) for educational attainment that extends our earlier discovery sample of 101,069 individuals to 293,723 individuals, and a replication study in an independent sample of 111,349 individuals from the UK Biobank. We identify 74 genome-wide significant loci associated with the number of years of schooling completed. Single-nucleotide polymorphisms associated with educational attainment are disproportionately found in genomic regions regulating gene expression in the fetal brain. Candidate genes are preferentially expressed in neural tissue, especially during the prenatal period, and enriched for biological pathways involved in neural development. Our findings demonstrate that, even for a behavioural phenotype that is mostly environmentally determined, a well-powered GWAS identifies replicable associated genetic variants that suggest biologically relevant pathways. Because educational attainment is measured in large numbers of individuals, it will continue to be useful as a proxy phenotype in efforts to characterize the genetic influences of related phenotypes, including cognition and neuropsychiatric diseases.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Educational attainment is strongly influenced by social and other environmental factors, but genetic factors are estimated to account for at least 20% of the variation across individuals. Here we report the results of a genome-wide association study (GWAS) for educational attainment that extends our earlier discovery sample of 101,069 individuals to 293,723 individuals, and a replication study in an independent sample of 111,349 individuals from the UK Biobank. We identify 74 genome-wide significant loci associated with the number of years of schooling completed. Single-nucleotide polymorphisms associated with educational attainment are disproportionately found in genomic regions regulating gene expression in the fetal brain. Candidate genes are preferentially expressed in neural tissue, especially during the prenatal period, and enriched for biological pathways involved in neural development. Our findings demonstrate that, even for a behavioural phenotype that is mostly environmentally determined, a well-powered GWAS identifies replicable associated genetic variants that suggest biologically relevant pathways. Because educational attainment is measured in large numbers of individuals, it will continue to be useful as a proxy phenotype in efforts to characterize the genetic influences of related phenotypes, including cognition and neuropsychiatric diseases. |
Di Pasqua LG; Berardo C; Rizzo V; Richelmi P; Croce AC; Vairetti M; Ferrigno A MCD diet-induced steatohepatitis is associated with alterations in asymmetric dimethylarginine (ADMA) and its transporters. Journal Article In: Molecular and Cellular Biochemistry, vol. 419, no 1, pp. 147-155, 2016. @article{%a1:%Y_268,
title = {MCD diet-induced steatohepatitis is associated with alterations in asymmetric dimethylarginine (ADMA) and its transporters.},
author = {{Di Pasqua LG} and Berardo C and Rizzo V and Richelmi P and Croce AC and Vairetti M and Ferrigno A},
url = {http://link.springer.com/article/10.1007%2Fs11010-016-2758-2},
doi = {10.1007/s11010-016-2758-2},
year = {2016},
date = {2016-02-27},
journal = {Molecular and Cellular Biochemistry},
volume = {419},
number = {1},
pages = {147-155},
abstract = {Using an experimental model of NASH induced by a methionine-choline-deficient (MCD) diet, we investigated whether changes occur in serum and tissue levels of asymmetric dimethylarginine (ADMA). Male Wistar rats underwent NASH induced by 8-week feeding with an MCD diet. Serum and hepatic biopsies at 2, 4 and 8 weeks were taken, and serum enzymes, ADMA and nitrate/nitrite (NOx), were evaluated. Hepatic biopsies were used for mRNA and protein expression analysis of dimethylarginine dimethylaminohydrolase-1 (DDAH-1) and protein methyltransferases (PRMT-1), enzymes involved in ADMA metabolism and synthesis, respectively, and ADMA transporters (CAT-1, CAT-2A and CAT-2B). Lipid peroxides (TBARS), glutathione, ATP/ADP and DDAH activity were quantified. An increase in serum AST and ALT was detected in MCD animals. A time-dependent decrease in serum and tissue ADMA and increase in mRNA expression of DDAH-1 and PRMT-1 as well as higher rates of mRNA expression of CAT-1 and lower rates of CAT-2A and CAT-2B were found after 8-week MCD diet. An increase in serum NOx and no changes in protein expression in DDAH-1 and CAT-1 and higher content in CAT-2 and PRMT-1 were found at 8 weeks. Hepatic DDAH activity decreased with a concomitant increase in oxidative stress, as demonstrated by high TBARS levels and low glutathione content. In conclusion, a decrease in serum and tissue ADMA levels in the MCD rats was found associated with a reduction in DDAH activity due to the marked oxidative stress observed. Changes in ADMA levels and its transporters are innovative factors in the onset and progression of hepatic alterations correlated with MCD diet-induced NASH.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Using an experimental model of NASH induced by a methionine-choline-deficient (MCD) diet, we investigated whether changes occur in serum and tissue levels of asymmetric dimethylarginine (ADMA). Male Wistar rats underwent NASH induced by 8-week feeding with an MCD diet. Serum and hepatic biopsies at 2, 4 and 8 weeks were taken, and serum enzymes, ADMA and nitrate/nitrite (NOx), were evaluated. Hepatic biopsies were used for mRNA and protein expression analysis of dimethylarginine dimethylaminohydrolase-1 (DDAH-1) and protein methyltransferases (PRMT-1), enzymes involved in ADMA metabolism and synthesis, respectively, and ADMA transporters (CAT-1, CAT-2A and CAT-2B). Lipid peroxides (TBARS), glutathione, ATP/ADP and DDAH activity were quantified. An increase in serum AST and ALT was detected in MCD animals. A time-dependent decrease in serum and tissue ADMA and increase in mRNA expression of DDAH-1 and PRMT-1 as well as higher rates of mRNA expression of CAT-1 and lower rates of CAT-2A and CAT-2B were found after 8-week MCD diet. An increase in serum NOx and no changes in protein expression in DDAH-1 and CAT-1 and higher content in CAT-2 and PRMT-1 were found at 8 weeks. Hepatic DDAH activity decreased with a concomitant increase in oxidative stress, as demonstrated by high TBARS levels and low glutathione content. In conclusion, a decrease in serum and tissue ADMA levels in the MCD rats was found associated with a reduction in DDAH activity due to the marked oxidative stress observed. Changes in ADMA levels and its transporters are innovative factors in the onset and progression of hepatic alterations correlated with MCD diet-induced NASH. |
Marioni RE; Ritchie SJ; Joshi PK; Hagenaars SP; Okbay A; Fischer K; Adams MJ; Hill WD; Davies G; et al nad; Biino G; et al; Benjamin DJ Genetic variants linked to education predict longevity. Journal Article In: Proceedings of the National Academy of Sciences of the United States of America, vol. 113, no 47, pp. 13366-13371, 2016. @article{%a1:%Y_298,
title = {Genetic variants linked to education predict longevity.},
author = {Marioni RE and Ritchie SJ and Joshi PK and Hagenaars SP and Okbay A and Fischer K and Adams MJ and Hill WD and Davies G and {et al} nad and Biino G and {et al} and Benjamin DJ},
url = {http://www.pnas.org/content/early/2016/10/25/1605334113.long},
doi = {10.1073/pnas.1605334113},
year = {2016},
date = {2016-02-27},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
volume = {113},
number = {47},
pages = {13366-13371},
abstract = {Educational attainment is associated with many health outcomes, including longevity. It is also known to be substantially heritable. Here, we used data from three large genetic epidemiology cohort studies (Generation Scotland, n = ∼17,000; UK Biobank, n = ∼115,000; and the Estonian Biobank, n = ∼6,000) to test whether education-linked genetic variants can predict lifespan length. We did so by using cohort members' polygenic profile score for education to predict their parents' longevity. Across the three cohorts, meta-analysis showed that a 1 SD higher polygenic education score was associated with ∼2.7% lower mortality risk for both mothers (total ndeaths = 79,702) and ∼2.4% lower risk for fathers (total ndeaths = 97,630). On average, the parents of offspring in the upper third of the polygenic score distribution lived 0.55 y longer compared with those of offspring in the lower third. Overall, these results indicate that the genetic contributions to educational attainment are useful in the prediction of human longevity.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Educational attainment is associated with many health outcomes, including longevity. It is also known to be substantially heritable. Here, we used data from three large genetic epidemiology cohort studies (Generation Scotland, n = ∼17,000; UK Biobank, n = ∼115,000; and the Estonian Biobank, n = ∼6,000) to test whether education-linked genetic variants can predict lifespan length. We did so by using cohort members' polygenic profile score for education to predict their parents' longevity. Across the three cohorts, meta-analysis showed that a 1 SD higher polygenic education score was associated with ∼2.7% lower mortality risk for both mothers (total ndeaths = 79,702) and ∼2.4% lower risk for fathers (total ndeaths = 97,630). On average, the parents of offspring in the upper third of the polygenic score distribution lived 0.55 y longer compared with those of offspring in the lower third. Overall, these results indicate that the genetic contributions to educational attainment are useful in the prediction of human longevity. |
Crespan E; Furrer A; Rösinger M; Bertoletti F; Mentegari E; Chiapparini G; Imhof R; Ziegler N; Sturla SJ; Hubscher U; van Loon B; Maga G Impact of ribonucleotide incorporation by DNA polymerases beta and lambda on oxidative base excision repair. Journal Article In: Nature Communications, vol. 7, pp. 10805, 2016. @article{%a1:%Y_263,
title = {Impact of ribonucleotide incorporation by DNA polymerases beta and lambda on oxidative base excision repair.},
author = {Crespan E and Furrer A and Rösinger M and Bertoletti F and Mentegari E and Chiapparini G and Imhof R and Ziegler N and Sturla SJ and Hubscher U and van Loon B and Maga G},
url = {http://www.nature.com/ncomms/2016/160226/ncomms10805/full/ncomms10805.html},
doi = {10.1038/ncomms10805},
year = {2016},
date = {2016-02-26},
journal = {Nature Communications},
volume = {7},
pages = {10805},
abstract = {Oxidative stress is a very frequent source of DNA damage. Many cellular DNA polymerases (Pols) can incorporate ribonucleotides (rNMPs) during DNA synthesis. However, whether oxidative stress-triggered DNA repair synthesis contributes to genomic rNMPs incorporation is so far not fully understood. Human specialized Pols beta and lamdda are the important enzymes involved in the oxidative stress tolerance, acting both in base excision repair and in translesion synthesis past the very frequent oxidative lesion 7,8-dihydro-8-oxoguanine (8-oxo-G). We found that Pol beta, to a greater extent than Pol lambda can incorporate rNMPs opposite normal bases or 8-oxo-G, and with a different fidelity. Further, the incorporation of rNMPs opposite 8-oxo-G delays repair by DNA glycosylases. Studies in Pol beta- and lambda-deficient cell extracts suggest that Pol beta levels can greatly affect rNMP incorporation opposite oxidative DNA lesions.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Oxidative stress is a very frequent source of DNA damage. Many cellular DNA polymerases (Pols) can incorporate ribonucleotides (rNMPs) during DNA synthesis. However, whether oxidative stress-triggered DNA repair synthesis contributes to genomic rNMPs incorporation is so far not fully understood. Human specialized Pols beta and lamdda are the important enzymes involved in the oxidative stress tolerance, acting both in base excision repair and in translesion synthesis past the very frequent oxidative lesion 7,8-dihydro-8-oxoguanine (8-oxo-G). We found that Pol beta, to a greater extent than Pol lambda can incorporate rNMPs opposite normal bases or 8-oxo-G, and with a different fidelity. Further, the incorporation of rNMPs opposite 8-oxo-G delays repair by DNA glycosylases. Studies in Pol beta- and lambda-deficient cell extracts suggest that Pol beta levels can greatly affect rNMP incorporation opposite oxidative DNA lesions. |
Klionsky DJ; Abdelmohsen K; Abe A; Scovassi AI; et al Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition) Journal Article In: Autophagy, vol. 12, no 1, pp. 222, 2016. @article{%a1:%Y_288,
title = {Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)},
author = {Klionsky DJ and Abdelmohsen K and Abe A and Scovassi AI and {et al}},
url = {http://www.tandfonline.com/doi/full/10.1080/15548627.2015.1100356},
doi = {10.1080/15548627.2015.1100356},
year = {2016},
date = {2016-02-26},
journal = {Autophagy},
volume = {12},
number = {1},
pages = {222},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
|
Barban N; Jansen R; de Vlaming R; Vaez A; Mandemakers JJ; Tropf FC; Shen X; Wilson JF; Chasman DI; Nolte IM; et al; Biino G; et al ; Koellinger PD; den Hoed M; Snieder H; Mills MC Genome-wide analysis identifies 12 loci influencing human reproductive behavior. Journal Article In: Nature Genetics, vol. 48, no 12, pp. 1462-1472, 2016. @article{%a1:%Y_250,
title = {Genome-wide analysis identifies 12 loci influencing human reproductive behavior.},
author = {Barban N and Jansen R and de Vlaming R and Vaez A and Mandemakers JJ and Tropf FC and Shen X and Wilson JF and Chasman DI and Nolte IM and {et al} and Biino G and {et al } and Koellinger PD and den Hoed M and Snieder H and Mills MC},
url = {http://www.nature.com/ng/journal/v48/n12/full/ng.3698.html},
doi = {10.1038/ng.3698},
year = {2016},
date = {2016-02-25},
journal = {Nature Genetics},
volume = {48},
number = {12},
pages = {1462-1472},
abstract = {The genetic architecture of human reproductive behavior-age at first birth (AFB) and number of children ever born (NEB)-has a strong relationship with fitness, human development, infertility and risk of neuropsychiatric disorders. However, very few genetic loci have been identified, and the underlying mechanisms of AFB and NEB are poorly understood. We report a large genome-wide association study of both sexes including 251,151 individuals for AFB and 343,072 individuals for NEB. We identified 12 independent loci that are significantly associated with AFB and/or NEB in a SNP-based genome-wide association study and 4 additional loci associated in a gene-based effort. These loci harbor genes that are likely to have a role, either directly or by affecting non-local gene expression, in human reproduction and infertility, thereby increasing understanding of these complex traits.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The genetic architecture of human reproductive behavior-age at first birth (AFB) and number of children ever born (NEB)-has a strong relationship with fitness, human development, infertility and risk of neuropsychiatric disorders. However, very few genetic loci have been identified, and the underlying mechanisms of AFB and NEB are poorly understood. We report a large genome-wide association study of both sexes including 251,151 individuals for AFB and 343,072 individuals for NEB. We identified 12 independent loci that are significantly associated with AFB and/or NEB in a SNP-based genome-wide association study and 4 additional loci associated in a gene-based effort. These loci harbor genes that are likely to have a role, either directly or by affecting non-local gene expression, in human reproduction and infertility, thereby increasing understanding of these complex traits. |
Dell'Orco M; Milani P; Arrigoni L; Pansarasa O; Sardone V; Maffioli E; Polveraccio F; Bordoni M; Diamanti L; Peverali FA; Tedeschi G; Cereda C Hydrogen peroxide-dependent oxidative stress induces SOD1 transcription gene is independent from Nrf2 transcription factor in a cellular model of neurodegeneration. Journal Article In: Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms, vol. 1859, no 2, pp. 315-323, 2016. @article{%a1:%Y_266,
title = {Hydrogen peroxide-dependent oxidative stress induces SOD1 transcription gene is independent from Nrf2 transcription factor in a cellular model of neurodegeneration.},
author = {{Dell'Orco M} and Milani P and Arrigoni L and Pansarasa O and Sardone V and Maffioli E and Polveraccio F and Bordoni M and Diamanti L and Peverali FA and Tedeschi G and Cereda C},
url = {http://www.sciencedirect.com/science/article/pii/S1874939915002485},
doi = {10.1016/j.bbagrm.2015.11.009},
year = {2016},
date = {2016-02-25},
journal = {Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms},
volume = {1859},
number = {2},
pages = {315-323},
abstract = {BACKGROUND: It is still unclear whether oxidative stress (OS) is a disease consequence or is directly involved in the etiology of neurodegenerative disorders (NDs) onset and/or progression; however, many of these conditions are associated with increased levels of oxidation markers and damaged cell components. Previously we demonstrated the accumulation of reactive oxygen species (ROS) and increased SOD1 gene expression within H2O2 SH-SY5Y treated cells recapitulating pathological features of Amyotrophic Lateral Sclerosis (ALS). Since we observed a post-transcriptional regulation of SOD1 gene in this cellular model of ALS, we investigated the transcriptional regulation of SOD1 mRNA under oxidative stress (OS). RESULTS: In response to H2O2 treatment, PolII increased its association to SOD1 promoter. Electrophoretic mobility shift assays (EMSA) and mass spectrometry analyses on SOD1 promoter highlighted the formation of a transcriptional complex bound to the ARE sequences. WB analyses showed that in our in vitro model, H2O2 exposure increases Nrf2 nuclear fraction while IP experiments confirmed its phosphorylation and release from Keap1 inhibition. However, H2O2 treatment did not modify Nrf2 binding on SOD1 promoter, which seems to be regulated by different TFs. CONCLUSIONS: Although our data suggest that SOD1 is transcriptionally regulated in response to OS, Nrf2 does not appear to associate with SOD1 promoter in this model of ALS. Our results open new perspectives in the comprehension of two key antioxidant pathways involved in neurodegeneration. Copyright 2015. Published by Elsevier B.V.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
BACKGROUND: It is still unclear whether oxidative stress (OS) is a disease consequence or is directly involved in the etiology of neurodegenerative disorders (NDs) onset and/or progression; however, many of these conditions are associated with increased levels of oxidation markers and damaged cell components. Previously we demonstrated the accumulation of reactive oxygen species (ROS) and increased SOD1 gene expression within H2O2 SH-SY5Y treated cells recapitulating pathological features of Amyotrophic Lateral Sclerosis (ALS). Since we observed a post-transcriptional regulation of SOD1 gene in this cellular model of ALS, we investigated the transcriptional regulation of SOD1 mRNA under oxidative stress (OS). RESULTS: In response to H2O2 treatment, PolII increased its association to SOD1 promoter. Electrophoretic mobility shift assays (EMSA) and mass spectrometry analyses on SOD1 promoter highlighted the formation of a transcriptional complex bound to the ARE sequences. WB analyses showed that in our in vitro model, H2O2 exposure increases Nrf2 nuclear fraction while IP experiments confirmed its phosphorylation and release from Keap1 inhibition. However, H2O2 treatment did not modify Nrf2 binding on SOD1 promoter, which seems to be regulated by different TFs. CONCLUSIONS: Although our data suggest that SOD1 is transcriptionally regulated in response to OS, Nrf2 does not appear to associate with SOD1 promoter in this model of ALS. Our results open new perspectives in the comprehension of two key antioxidant pathways involved in neurodegeneration. Copyright 2015. Published by Elsevier B.V. |
Fassihi H; Sethi M; Fawcett H; Wing J; Chandler N; Mohammed S; Craythorne E; Morley AM; Lim R; Turner S; Henshaw T; Garrood I; Giunti P; Hedderly T; Abiona A; Naik H; Harrop G; McGibbon D; Jaspers NG; Botta E; Nardo T; Stefanini M; Young AR; Sarkany RP; Lehmann AR Deep phenotyping of 89 xeroderma pigmentosum patients reveals unexpected heterogeneity dependent on the precise molecular defect. Journal Article In: Proceedings of the National Academy of Sciences of the United States of America, vol. 113, no 9, pp. 1236-1245, 2016. @article{%a1:%Y_278,
title = {Deep phenotyping of 89 xeroderma pigmentosum patients reveals unexpected heterogeneity dependent on the precise molecular defect.},
author = {Fassihi H and Sethi M and Fawcett H and Wing J and Chandler N and Mohammed S and Craythorne E and Morley AM and Lim R and Turner S and Henshaw T and Garrood I and Giunti P and Hedderly T and Abiona A and Naik H and Harrop G and McGibbon D and Jaspers NG and Botta E and Nardo T and Stefanini M and Young AR and Sarkany RP and Lehmann AR},
url = {http://www.pnas.org/content/113/9/E1236.long},
doi = {10.1073/pnas.1519444113},
year = {2016},
date = {2016-02-25},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
volume = {113},
number = {9},
pages = {1236-1245},
abstract = {Xeroderma pigmentosum (XP) is a rare DNA repair disorder characterized by increased susceptibility to UV radiation (UVR)-induced skin pigmentation, skin cancers, ocular surface disease, and, in some patients, sunburn and neurological degeneration. Genetically, it is assigned to eight complementation groups (XP-A to -G and variant). For the last 5 y, the UK national multidisciplinary XP service has provided follow-up for 89 XP patients, representing most of the XP patients in the United Kingdom. Causative mutations, DNA repair levels, and more than 60 clinical variables relating to dermatology, ophthalmology, and neurology have been measured, using scoring systems to categorize disease severity. This deep phenotyping has revealed unanticipated heterogeneity of clinical features, between and within complementation groups. Skin cancer is most common in XP-C, XP-E, and XP-V patients, previously considered to be the milder groups based on cellular analyses. These patients have normal sunburn reactions and are therefore diagnosed later and are less likely to adhere to UVR protection. XP-C patients are specifically hypersensitive to ocular damage, and XP-F and XP-G patients appear to be much less susceptible to skin cancer than other XP groups. Within XP groups, different mutations confer susceptibility or resistance to neurological damage. Our findings on this large cohort of XP patients under long-term follow-up reveal that XP is more heterogeneous than has previously been appreciated. Our data now enable provision of personalized prognostic information and management advice for each XP patient, as well as providing new insights into the functions of the XP proteins.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Xeroderma pigmentosum (XP) is a rare DNA repair disorder characterized by increased susceptibility to UV radiation (UVR)-induced skin pigmentation, skin cancers, ocular surface disease, and, in some patients, sunburn and neurological degeneration. Genetically, it is assigned to eight complementation groups (XP-A to -G and variant). For the last 5 y, the UK national multidisciplinary XP service has provided follow-up for 89 XP patients, representing most of the XP patients in the United Kingdom. Causative mutations, DNA repair levels, and more than 60 clinical variables relating to dermatology, ophthalmology, and neurology have been measured, using scoring systems to categorize disease severity. This deep phenotyping has revealed unanticipated heterogeneity of clinical features, between and within complementation groups. Skin cancer is most common in XP-C, XP-E, and XP-V patients, previously considered to be the milder groups based on cellular analyses. These patients have normal sunburn reactions and are therefore diagnosed later and are less likely to adhere to UVR protection. XP-C patients are specifically hypersensitive to ocular damage, and XP-F and XP-G patients appear to be much less susceptible to skin cancer than other XP groups. Within XP groups, different mutations confer susceptibility or resistance to neurological damage. Our findings on this large cohort of XP patients under long-term follow-up reveal that XP is more heterogeneous than has previously been appreciated. Our data now enable provision of personalized prognostic information and management advice for each XP patient, as well as providing new insights into the functions of the XP proteins. |
Zanoni M; Piccinini F; Arienti C; Zamagni A; Santi S; Polico R; Bevilacqua A; Tesei A 3D tumor spheroid models for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained. Journal Article In: Scientific Reports, vol. 6, pp. 19103, 2016. @article{%a1:%Y_318,
title = {3D tumor spheroid models for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained.},
author = {Zanoni M and Piccinini F and Arienti C and Zamagni A and Santi S and Polico R and Bevilacqua A and Tesei A},
url = {http://www.nature.com/articles/srep19103},
doi = {10.1038/srep19103},
year = {2016},
date = {2016-02-25},
journal = {Scientific Reports},
volume = {6},
pages = {19103},
abstract = {The potential of a spheroid tumor model composed of cells in different proliferative and metabolic states for the development of new anticancer strategies has been amply demonstrated. However, there is little or no information in the literature on the problems of reproducibility of data originating from experiments using 3D models. Our analyses, carried out using a novel open source software capable of performing an automatic image analysis of 3D tumor colonies, showed that a number of morphology parameters affect the response of large spheroids to treatment. In particular, we found that both spheroid volume and shape may be a source of variability. We also compared some commercially available viability assays specifically designed for 3D models. In conclusion, our data indicate the need for a pre-selection of tumor spheroids of homogeneous volume and shape to reduce data variability to a minimum before use in a cytotoxicity test. In addition, we identified and validated a cytotoxicity test capable of providing meaningful data on the damage induced in large tumor spheroids of up to diameter in 650 μm by different kinds of treatments.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The potential of a spheroid tumor model composed of cells in different proliferative and metabolic states for the development of new anticancer strategies has been amply demonstrated. However, there is little or no information in the literature on the problems of reproducibility of data originating from experiments using 3D models. Our analyses, carried out using a novel open source software capable of performing an automatic image analysis of 3D tumor colonies, showed that a number of morphology parameters affect the response of large spheroids to treatment. In particular, we found that both spheroid volume and shape may be a source of variability. We also compared some commercially available viability assays specifically designed for 3D models. In conclusion, our data indicate the need for a pre-selection of tumor spheroids of homogeneous volume and shape to reduce data variability to a minimum before use in a cytotoxicity test. In addition, we identified and validated a cytotoxicity test capable of providing meaningful data on the damage induced in large tumor spheroids of up to diameter in 650 μm by different kinds of treatments. |
Castagnaro S; Pellegrini C; Pellegrini M; Chrisam M; Sabatelli P; Toni S; Grumati P; Ripamonti C; Pratelli L; Maraldi NM; Cocchi D; Righi V; Faldini C; Sandri M; Bonaldo P; Merlini L Autophagy activation in COL6 myopathic patients by a low-protein-diet pilot trial. Journal Article In: Autophagy, vol. 12, no 12, pp. 2484-2495, 2016. @article{%a1:%Y_258,
title = {Autophagy activation in COL6 myopathic patients by a low-protein-diet pilot trial.},
author = {Castagnaro S and Pellegrini C and Pellegrini M and Chrisam M and Sabatelli P and Toni S and Grumati P and Ripamonti C and Pratelli L and Maraldi NM and Cocchi D and Righi V and Faldini C and Sandri M and Bonaldo P and Merlini L},
url = {http://www.tandfonline.com/doi/full/10.1080/15548627.2016.1231279},
doi = {10.1080/15548627.2016.1231279},
year = {2016},
date = {2016-02-24},
journal = {Autophagy},
volume = {12},
number = {12},
pages = {2484-2495},
abstract = {A pilot clinical trial based on nutritional modulation was designed to assess the efficacy of a one-year low-protein diet in activating autophagy in skeletal muscle of patients affected by COL6/collagen VI-related myopathies. Ullrich congenital muscular dystrophy and Bethlem myopathy are rare inherited muscle disorders caused by mutations of COL6 genes and for which no cure is yet available. Studies in col6 null mice revealed that myofiber degeneration involves autophagy defects and that forced activation of autophagy results in the amelioration of muscle pathology. Seven adult patients affected by COL6 myopathies underwent a controlled low-protein diet for 12 mo and we evaluated the presence of autophagosomes and the mRNA and protein levels for BECN1/Beclin 1 and MAP1LC3B/LC3B in muscle biopsies and blood leukocytes. Safety measures were assessed, including muscle strength, motor and respiratory function, and metabolic parameters. After one y of low-protein diet, autophagic markers were increased in skeletal muscle and blood leukocytes of patients. The treatment was safe as shown by preservation of lean:fat percentage of body composition, muscle strength and function. Moreover, the decreased incidence of myofiber apoptosis indicated benefits in muscle homeostasis, and the metabolic changes pointed at improved mitochondrial function. These data provide evidence that a low-protein diet is able to activate autophagy and is safe and tolerable in patients with COL6 myopathies, pointing at autophagy activation as a potential target for therapeutic applications. In addition, our findings indicate that blood leukocytes are a promising noninvasive tool for monitoring autophagy activation in patients.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
A pilot clinical trial based on nutritional modulation was designed to assess the efficacy of a one-year low-protein diet in activating autophagy in skeletal muscle of patients affected by COL6/collagen VI-related myopathies. Ullrich congenital muscular dystrophy and Bethlem myopathy are rare inherited muscle disorders caused by mutations of COL6 genes and for which no cure is yet available. Studies in col6 null mice revealed that myofiber degeneration involves autophagy defects and that forced activation of autophagy results in the amelioration of muscle pathology. Seven adult patients affected by COL6 myopathies underwent a controlled low-protein diet for 12 mo and we evaluated the presence of autophagosomes and the mRNA and protein levels for BECN1/Beclin 1 and MAP1LC3B/LC3B in muscle biopsies and blood leukocytes. Safety measures were assessed, including muscle strength, motor and respiratory function, and metabolic parameters. After one y of low-protein diet, autophagic markers were increased in skeletal muscle and blood leukocytes of patients. The treatment was safe as shown by preservation of lean:fat percentage of body composition, muscle strength and function. Moreover, the decreased incidence of myofiber apoptosis indicated benefits in muscle homeostasis, and the metabolic changes pointed at improved mitochondrial function. These data provide evidence that a low-protein diet is able to activate autophagy and is safe and tolerable in patients with COL6 myopathies, pointing at autophagy activation as a potential target for therapeutic applications. In addition, our findings indicate that blood leukocytes are a promising noninvasive tool for monitoring autophagy activation in patients. |
Cavallo C; Desando G; Ferrari A; Zini N; Mariani E; Grigolo B Hyaluronan scaffold supports osteogenic differentiation of bone marrow concentrate cells. Journal Article In: Journal of Biological Regulators and Homeostatic Agents, vol. 30, no 2, pp. 409-420, 2016. @article{%a1:%Y_259,
title = {Hyaluronan scaffold supports osteogenic differentiation of bone marrow concentrate cells.},
author = {Cavallo C and Desando G and Ferrari A and Zini N and Mariani E and Grigolo B},
url = {https://www.biolifesas.org/biolife/jbrha-2/},
year = {2016},
date = {2016-02-20},
journal = {Journal of Biological Regulators and Homeostatic Agents},
volume = {30},
number = {2},
pages = {409-420},
abstract = {Osteochondral lesions are considered a challenge for orthopedic surgeons. Currently, the treatments available are often unsatisfactory and unable to stimulate tissue regeneration. Tissue engineering offers a new therapeutic strategy, taking into account the role exerted by cells, biomaterial and growth factors in restoring tissue damage. In this light, Mesenchymal Stem Cells (MSCs) have been indicated as a fascinating tool for regenerative medicine thanks to their ability to differentiate into bone, cartilage and adipose tissue. However, in vitro-cultivation of MSCs could be associated with some risks such as de-differentiation/reprogramming, infection and contaminations of the cells. To overcome these shortcomings, a new approach is represented by the use of Bone Marrow Concentrate (BMC), that could allow the delivery of cells surrounded by their microenvironment in injured tissue. For this purpose, cells require a tridimensional scaffold that can support their adhesion, proliferation and differentiation. This study is focused on the potentiality of BMC seeded onto a hyaluronan-based scaffold (Hyaff-11) to differentiate into osteogenic lineage. This process depends on the specific interaction between cells derived from bone marrow (surrounded by their niche) and scaffold, that create an environment able to support the regeneration of damaged tissue. The data obtained from the present study demonstrate that BMC grown onto Hyaff-11 are able to differentiate toward osteogenic sense, producing specific osteogenic genes and matrix proteins.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Osteochondral lesions are considered a challenge for orthopedic surgeons. Currently, the treatments available are often unsatisfactory and unable to stimulate tissue regeneration. Tissue engineering offers a new therapeutic strategy, taking into account the role exerted by cells, biomaterial and growth factors in restoring tissue damage. In this light, Mesenchymal Stem Cells (MSCs) have been indicated as a fascinating tool for regenerative medicine thanks to their ability to differentiate into bone, cartilage and adipose tissue. However, in vitro-cultivation of MSCs could be associated with some risks such as de-differentiation/reprogramming, infection and contaminations of the cells. To overcome these shortcomings, a new approach is represented by the use of Bone Marrow Concentrate (BMC), that could allow the delivery of cells surrounded by their microenvironment in injured tissue. For this purpose, cells require a tridimensional scaffold that can support their adhesion, proliferation and differentiation. This study is focused on the potentiality of BMC seeded onto a hyaluronan-based scaffold (Hyaff-11) to differentiate into osteogenic lineage. This process depends on the specific interaction between cells derived from bone marrow (surrounded by their niche) and scaffold, that create an environment able to support the regeneration of damaged tissue. The data obtained from the present study demonstrate that BMC grown onto Hyaff-11 are able to differentiate toward osteogenic sense, producing specific osteogenic genes and matrix proteins. |
Lemma S; Sboarina M; Porporato PE; Zini N; Sonveaux P; Di Pompo G; Baldini N; Avnet S Energy metabolism in osteoclast formation and activity. Journal Article In: International Journal of Biochemistry And Cell Biology, vol. 79, pp. 168-180, 2016. @article{%a1:%Y_291,
title = {Energy metabolism in osteoclast formation and activity.},
author = {Lemma S and Sboarina M and Porporato PE and Zini N and Sonveaux P and Di Pompo G and Baldini N and Avnet S},
url = {10.1016/j.biocel.2016.08.034},
doi = {10.1016/j.biocel.2016.08.034},
year = {2016},
date = {2016-02-20},
journal = {International Journal of Biochemistry And Cell Biology},
volume = {79},
pages = {168-180},
abstract = {Osteoclastogenesis and osteolysis are energy-consuming processes supported by high metabolic activities. In human osteoclasts derived from the fusion of monocytic precursors, we found a substantial increase in the number of mitochondria with differentiation. In mature osteoclasts, mitochondria were also increased in size, rich of cristae and arranged in a complex tubular network. When compared with immature cells, fully differentiated osteoclasts showed higher levels of enzymes of the electron transport chain, a higher mitochondrial oxygen consumption rate and a lower glycolytic efficiency, as evaluated by extracellular flux analysis and by the quantification of metabolites in the culture supernatant. Thus, oxidative phosphorylation appeared the main bioenergetic source for osteoclast formation. Conversely, we found that bone resorption mainly relied on glycolysis. In fact, osteoclast fuelling with galactose, forcing cells to depend on Oxidative Phosphorylation by reducing the rate of glycolysis, significantly impaired Type I collagen degradation, whereas non-cytotoxic doses of rotenone, an inhibitor of the mitochondrial complex I, enhanced osteoclast activity. Furthermore, we found that the enzymes associated to the glycolytic pathway are localised close to the actin ring of polarised osteoclasts, where energy-demanding activities associated with bone degradation take place. In conclusion, we demonstrate that the energy required for osteoclast differentiation mainly derives from mitochondrial oxidative metabolism, whereas the peripheral cellular activities associated with bone matrix degradation are supported by glycolysis. A better understanding of human osteoclast energy metabolism holds the potential for future therapeutic interventions aimed to target osteoclast activity in different pathological conditions of bone.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Osteoclastogenesis and osteolysis are energy-consuming processes supported by high metabolic activities. In human osteoclasts derived from the fusion of monocytic precursors, we found a substantial increase in the number of mitochondria with differentiation. In mature osteoclasts, mitochondria were also increased in size, rich of cristae and arranged in a complex tubular network. When compared with immature cells, fully differentiated osteoclasts showed higher levels of enzymes of the electron transport chain, a higher mitochondrial oxygen consumption rate and a lower glycolytic efficiency, as evaluated by extracellular flux analysis and by the quantification of metabolites in the culture supernatant. Thus, oxidative phosphorylation appeared the main bioenergetic source for osteoclast formation. Conversely, we found that bone resorption mainly relied on glycolysis. In fact, osteoclast fuelling with galactose, forcing cells to depend on Oxidative Phosphorylation by reducing the rate of glycolysis, significantly impaired Type I collagen degradation, whereas non-cytotoxic doses of rotenone, an inhibitor of the mitochondrial complex I, enhanced osteoclast activity. Furthermore, we found that the enzymes associated to the glycolytic pathway are localised close to the actin ring of polarised osteoclasts, where energy-demanding activities associated with bone degradation take place. In conclusion, we demonstrate that the energy required for osteoclast differentiation mainly derives from mitochondrial oxidative metabolism, whereas the peripheral cellular activities associated with bone matrix degradation are supported by glycolysis. A better understanding of human osteoclast energy metabolism holds the potential for future therapeutic interventions aimed to target osteoclast activity in different pathological conditions of bone. |
Montecucco A; Biamonti G DNA and RNA metabolism meet at chromatin to control genome stability Journal Article In: Frontiers in Genetics, vol. 7, pp. 67, 2016. @article{%a1:%Y_300,
title = {DNA and RNA metabolism meet at chromatin to control genome stability},
author = {Montecucco A and Biamonti G},
url = {http://journal.frontiersin.org/article/10.3389/fgene.2016.00067/full},
doi = {doi: 10.3389/fgene.2016.00067},
year = {2016},
date = {2016-02-19},
journal = {Frontiers in Genetics},
volume = {7},
pages = {67},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
|
Scovassi AI Exosomes: Extracellular vesicles transporting macromolecules Journal Article In: Biochemistry & Pharmacology: Open Access, vol. 5, pp. e181, 2016. @article{%a1:%Y_311,
title = {Exosomes: Extracellular vesicles transporting macromolecules},
author = {Scovassi AI},
url = {http://www.omicsgroup.org/journals/exosomes-extracellular-vesicles-transporting-macromolecules-2167-0501-1000e181.php?aid=70857},
doi = { 10.4172/2167-0501.1000e181},
year = {2016},
date = {2016-02-19},
journal = {Biochemistry & Pharmacology: Open Access},
volume = {5},
pages = {e181},
abstract = {Extracellular vesicles (EVs), firstly identified in 1983 in the conditioned culture medium collected during the maturation of reticulocytes into erythrocytes, are released by the cells into the extracellular milieu. EVs are present in all human body fluids; their number and content are often modulated in pathological samples, thus making interesting their evaluation as possible disease biomarkers.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Extracellular vesicles (EVs), firstly identified in 1983 in the conditioned culture medium collected during the maturation of reticulocytes into erythrocytes, are released by the cells into the extracellular milieu. EVs are present in all human body fluids; their number and content are often modulated in pathological samples, thus making interesting their evaluation as possible disease biomarkers. |
Barteri M; De Carolis R; Marinelli F; Tomassetti G; Montemiglio LC Effects of microwaves (900 MHz) on peroxidase systems: a comparison between lactoperoxidase and horseradish peroxidase. Journal Article In: Electromagnetic Biology and Medicine, vol. 35, no 2, pp. 126-133, 2016. @article{%a1:%Y_251,
title = {Effects of microwaves (900 MHz) on peroxidase systems: a comparison between lactoperoxidase and horseradish peroxidase.},
author = {Barteri M and De Carolis R and Marinelli F and Tomassetti G and Montemiglio LC},
url = {http://www.tandfonline.com/doi/abs/10.3109/15368378.2014.1002135?journalCode=iebm20},
doi = {10.3109/15368378.2014.1002135},
year = {2016},
date = {2016-02-18},
journal = {Electromagnetic Biology and Medicine},
volume = {35},
number = {2},
pages = {126-133},
abstract = {This work shows the effects of exposure to an electromagnetic field at 900 MHz on the catalytic activity of the enzymes lactoperoxidase (LPO) and horseradish peroxidase (HRP). Experimental evidence that irradiation causes conformational changes of the active sites and influences the formation and stability of the intermediate free radicals is documented by measurements of enzyme kinetics, circular dichroism spectroscopy (CD) and cyclic voltammetry.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
This work shows the effects of exposure to an electromagnetic field at 900 MHz on the catalytic activity of the enzymes lactoperoxidase (LPO) and horseradish peroxidase (HRP). Experimental evidence that irradiation causes conformational changes of the active sites and influences the formation and stability of the intermediate free radicals is documented by measurements of enzyme kinetics, circular dichroism spectroscopy (CD) and cyclic voltammetry. |
Francia S; Cabrini M; Matti V; Oldani A; d'Adda di Fagagna F DICER, DROSHA and DNA damage response RNAs are necessary for the secondary recruitment of DNA damage response factors. Journal Article In: Journal of Cell Science, vol. 129, pp. 1468-1476, 2016. @article{%a1:%Y_280,
title = {DICER, DROSHA and DNA damage response RNAs are necessary for the secondary recruitment of DNA damage response factors.},
author = {Francia S and Cabrini M and Matti V and Oldani A and {d'Adda di Fagagna F}},
url = {http://jcs.biologists.org/content/129/7/1468.long},
doi = {10.1242/jcs.182188},
year = {2016},
date = {2016-02-18},
journal = {Journal of Cell Science},
volume = {129},
pages = {1468-1476},
abstract = {The DNA damage response (DDR) plays a central role in preserving genome integrity. Recently, we reported that the endoribonucleases DICER and DROSHA contribute to DDR activation by generating small non-coding RNAs, termed DNA damage response RNA (DDRNA), carrying the sequence of the damaged locus. It is presently unclear whether DDRNAs act by promoting the primary recognition of DNA lesions or the secondary recruitment of DDR factors into cytologically detectable foci and consequent signal amplification. Here, we demonstrate that DICER and DROSHA are dispensable for primary recruitment of the DDR sensor NBS1 to DNA damage sites. Instead, the accumulation of the DDR mediators MDC1 and 53BP1 (also known as TP53BP1), markers of secondary recruitment, is reduced in DICER- or DROSHA-inactivated cells. In addition, NBS1 (also known as NBN) primary recruitment is resistant to RNA degradation, consistent with the notion that RNA is dispensable for primary recognition of DNA lesions. We propose that DICER, DROSHA and DDRNAs act in the response to DNA damage after primary recognition of DNA lesions and, together with γH2AX, are essential for enabling the secondary recruitment of DDR factors and fuel the amplification of DDR signaling. 2016. Published by The Company of Biologists Ltd.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The DNA damage response (DDR) plays a central role in preserving genome integrity. Recently, we reported that the endoribonucleases DICER and DROSHA contribute to DDR activation by generating small non-coding RNAs, termed DNA damage response RNA (DDRNA), carrying the sequence of the damaged locus. It is presently unclear whether DDRNAs act by promoting the primary recognition of DNA lesions or the secondary recruitment of DDR factors into cytologically detectable foci and consequent signal amplification. Here, we demonstrate that DICER and DROSHA are dispensable for primary recruitment of the DDR sensor NBS1 to DNA damage sites. Instead, the accumulation of the DDR mediators MDC1 and 53BP1 (also known as TP53BP1), markers of secondary recruitment, is reduced in DICER- or DROSHA-inactivated cells. In addition, NBS1 (also known as NBN) primary recruitment is resistant to RNA degradation, consistent with the notion that RNA is dispensable for primary recognition of DNA lesions. We propose that DICER, DROSHA and DDRNAs act in the response to DNA damage after primary recognition of DNA lesions and, together with γH2AX, are essential for enabling the secondary recruitment of DDR factors and fuel the amplification of DDR signaling. 2016. Published by The Company of Biologists Ltd. |
Gallorini M; di Giacomo V; Di Valerio V; Rapino M; Bosco D; Travan A; Di Giulio M; Di Pietro R; Paoletti S; Cataldi A; Sancilio S Cell-protection mechanism through autophagy in HGFs/S. mitis co-culture treated with Chitlac-nAg. Journal Article In: Journal of Materials Science. Materials in Medicine., vol. 27, no 12, pp. 186, 2016. @article{%a1:%Y_281,
title = {Cell-protection mechanism through autophagy in HGFs/S. mitis co-culture treated with Chitlac-nAg.},
author = {Gallorini M and di Giacomo V and Di Valerio V and Rapino M and Bosco D and Travan A and Di Giulio M and Di Pietro R and Paoletti S and Cataldi A and Sancilio S},
url = {http://link.springer.com/article/10.1007%2Fs10856-016-5803-5},
doi = {10.1007/s10856-016-5803-5},
year = {2016},
date = {2016-02-18},
journal = {Journal of Materials Science. Materials in Medicine.},
volume = {27},
number = {12},
pages = {186},
abstract = {Silver-based products have been proven to be effective in retarding and preventing bacterial growth since ancient times. In the field of restorative dentistry, the use of silver ions/nanoparticles has been explored to counteract bacterial infections, as silver can destroy bacterial cell walls by reacting with membrane proteins. However, it is also cytotoxic towards eukaryotic cells, which are capable of internalizing nanoparticles. In this work, we investigated the biological effects of Chitlac-nAg, a colloidal system based on a modified chitosan (Chitlac), administered for 24-48 h to a co-culture of primary human gingival fibroblasts and Streptococcus mitis in the presence of saliva, developed to mimic the microenvironment of the oral cavity. We sought to determine its efficiency to combat oral hygiene-related diseases without affecting eukaryotic cells. Cytotoxicity, reactive oxygen species production, apoptosis induction, nanoparticles uptake, and lysosome and autophagosome metabolism were evaluated. In vitro results show that Chitlac-nAg does not exert cytotoxic effects on human gingival fibroblasts, which seem to survive through a homoeostasis mechanism involving autophagy. That suggests that the novel biomaterial Chitlac-nAg could be a promising tool in the field of dentistry.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Silver-based products have been proven to be effective in retarding and preventing bacterial growth since ancient times. In the field of restorative dentistry, the use of silver ions/nanoparticles has been explored to counteract bacterial infections, as silver can destroy bacterial cell walls by reacting with membrane proteins. However, it is also cytotoxic towards eukaryotic cells, which are capable of internalizing nanoparticles. In this work, we investigated the biological effects of Chitlac-nAg, a colloidal system based on a modified chitosan (Chitlac), administered for 24-48 h to a co-culture of primary human gingival fibroblasts and Streptococcus mitis in the presence of saliva, developed to mimic the microenvironment of the oral cavity. We sought to determine its efficiency to combat oral hygiene-related diseases without affecting eukaryotic cells. Cytotoxicity, reactive oxygen species production, apoptosis induction, nanoparticles uptake, and lysosome and autophagosome metabolism were evaluated. In vitro results show that Chitlac-nAg does not exert cytotoxic effects on human gingival fibroblasts, which seem to survive through a homoeostasis mechanism involving autophagy. That suggests that the novel biomaterial Chitlac-nAg could be a promising tool in the field of dentistry. |
Goodson WH 3rd; Lowe L; Carpenter DO; Gilbertson M; Manaf Ali A; Lopez de Cerain Salsamendi A; Lasfar A; Carnero A; Azqueta A; Amedei A; Charles AK; Collins AR; Ward A; Salzberg AC; Colacci A; Olsen AK; Berg A; Barclay BJ; Zhou BP; Blanco-Aparicio C; Baglole CJ; Dong C; Mondello C; Hsu CW; Naus CC; Yedjou C; Curran CS; Laird DW; Koch DC; Carlin DJ; Felsher DW; Roy D; Brown DG; Ratovitski E; Ryan EP; Corsini E; Rojas E; Moon EY; Laconi E; Marongiu F; Al-Mulla F; Chiaradonna F; Darroudi F; Martin FL; Van Schooten FJ; Goldberg GS; Wagemaker G; Nangami G; Calaf GM; Williams G; Wolf GT; Koppen G; Brunborg G; Kim Lyerly H; Krishnan H; Ab Hamid H; Yasaei H; Sone H; Kondoh H; Salem HK; Hsu HY; Park HH; Koturbash I; Miousse IR; Scovassi AI; Klaunig JE; Vondráček J; Raju J; Roman J; Wise JP Sr; Whitfield JR; Woodrick J; Christopher JA; Ochieng J; Martinez-Leal JF; Weisz J; Kravchenko J; Sun J; Prudhomme KR; Narayanan KB; Cohen-Solal KA; Moorwood K; Gonzalez L; Soucek L; Jian L; D'Abronzo LS; Lin LT; Li L; Gulliver L; McCawley LJ; Memeo L; Vermeulen L; Leyns L; Zhang L; Valverde M; Khatami M; Romano MF; Chapellier M; Williams MA; Wade M; Manjili MH; Lleonart M; Xia M; Gonzalez MJ; Karamouzis MV; Kirsch-Volders M; Vaccari M; Kuemmerle NB; Singh N; Cruickshanks N; Kleinstreuer N; van Larebeke N; Ahmed N; Ogunkua O; Krishnakumar PK; Vadgama P; Marignani PA; Ghosh PM; Ostrosky-Wegman P; Thompson P; Dent P; Heneberg P; Darbre P; Sing Leung P; Nangia-Makker P; Cheng QS; Robey RB; Al-Temaimi R; Roy R; Andrade-Vieira R; Sinha RK; Mehta R; Vento R; Di Fiore R; Ponce-Cusi R; Dornetshuber-Fleiss R; Nahta R; Castellino RC; Palorini R; Abd Hamid R; Langie SA; Eltom S; Brooks SA; Ryeom S; Wise SS; Bay SN; Harris SA; Papagerakis S; Romano S; Pavanello S; Eriksson S; Forte S; Casey SC; Luanpitpong S; Lee TJ; Otsuki T; Chen T; Massfelder T; Sanderson T; Guarnieri T; Hultman T; Dormoy V; Odero-Marah V; Sabbisetti V; Maguer-Satta V; Rathmell WK; Engström W; Decker WK; Bisson WH; Rojanasakul Y; Luqmani Y; Chen Z; Hu Z Llona-Minguez S; Hoglund A; Jacques SA; Johansson L; Calderon-Montano JM; Claesson M; Loseva O; Valerie NC; Lundbäck T; Piedrafita J; Maga G; Crespan E; Meijer L; Burgos Morón E; Baranczewski P; Hagbjork AL; Svensson R; Wiita E; Almlof I; Visnes T; Jeppsson F; Sigmundsson K; Jensen AJ; Artursson P; Jemth AS; Stenmark P; Warpman Berglund U; Scobie M; Helleday T Discovery of the First Potent and Selective Inhibitors of Human dCTP Pyrophosphatase 1. Journal Article In: Journal of medicinal chemistry, vol. 59, no 3, pp. 1140-1148, 2016. @article{%a1:%Y_292,
title = {Discovery of the First Potent and Selective Inhibitors of Human dCTP Pyrophosphatase 1.},
author = {{Goodson WH 3rd} and Lowe L and Carpenter DO and Gilbertson M and Manaf Ali A and Lopez de Cerain Salsamendi A and Lasfar A and Carnero A and Azqueta A and Amedei A and Charles AK and Collins AR and Ward A and Salzberg AC and Colacci A and Olsen AK and Berg A and Barclay BJ and Zhou BP and Blanco-Aparicio C and Baglole CJ and Dong C and Mondello C and Hsu CW and Naus CC and Yedjou C and Curran CS and Laird DW and Koch DC and Carlin DJ and Felsher DW and Roy D and Brown DG and Ratovitski E and Ryan EP and Corsini E and Rojas E and Moon EY and Laconi E and Marongiu F and Al-Mulla F and Chiaradonna F and Darroudi F and Martin FL and Van Schooten FJ and Goldberg GS and Wagemaker G and Nangami G and Calaf GM and Williams G and Wolf GT and Koppen G and Brunborg G and Kim Lyerly H and Krishnan H and Ab Hamid H and Yasaei H and Sone H and Kondoh H and Salem HK and Hsu HY and Park HH and Koturbash I and Miousse IR and Scovassi AI and Klaunig JE and Vondráček J and Raju J and Roman J and Wise JP Sr and Whitfield JR and Woodrick J and Christopher JA and Ochieng J and Martinez-Leal JF and Weisz J and Kravchenko J and Sun J and Prudhomme KR and Narayanan KB and Cohen-Solal KA and Moorwood K and Gonzalez L and Soucek L and Jian L and D'Abronzo LS and Lin LT and Li L and Gulliver L and McCawley LJ and Memeo L and Vermeulen L and Leyns L and Zhang L and Valverde M and Khatami M and Romano MF and Chapellier M and Williams MA and Wade M and Manjili MH and Lleonart M and Xia M and Gonzalez MJ and Karamouzis MV and Kirsch-Volders M and Vaccari M and Kuemmerle NB and Singh N and Cruickshanks N and Kleinstreuer N and van Larebeke N and Ahmed N and Ogunkua O and Krishnakumar PK and Vadgama P and Marignani PA and Ghosh PM and Ostrosky-Wegman P and Thompson P and Dent P and Heneberg P and Darbre P and Sing Leung P and Nangia-Makker P and Cheng QS and Robey RB and Al-Temaimi R and Roy R and Andrade-Vieira R and Sinha RK and Mehta R and Vento R and Di Fiore R and Ponce-Cusi R and Dornetshuber-Fleiss R and Nahta R and Castellino RC and Palorini R and Abd Hamid R and Langie SA and Eltom S and Brooks SA and Ryeom S and Wise SS and Bay SN and Harris SA and Papagerakis S and Romano S and Pavanello S and Eriksson S and Forte S and Casey SC and Luanpitpong S and Lee TJ and Otsuki T and Chen T and Massfelder T and Sanderson T and Guarnieri T and Hultman T and Dormoy V and Odero-Marah V and Sabbisetti V and Maguer-Satta V and Rathmell WK and Engström W and Decker WK and Bisson WH and Rojanasakul Y and Luqmani Y and Chen Z and Hu Z {Llona-Minguez S} and Hoglund A and Jacques SA and Johansson L and Calderon-Montano JM and Claesson M and Loseva O and Valerie NC and Lundbäck T and Piedrafita J and Maga G and Crespan E and Meijer L and Burgos Morón E and Baranczewski P and Hagbjork AL and Svensson R and Wiita E and Almlof I and Visnes T and Jeppsson F and Sigmundsson K and Jensen AJ and Artursson P and Jemth AS and Stenmark P and Warpman Berglund U and Scobie M and Helleday T},
url = {http://pubs.acs.org/doi/abs/10.1021/acs.jmedchem.5b01741},
doi = {10.1021/acs.jmedchem.5b01741},
year = {2016},
date = {2016-02-18},
journal = {Journal of medicinal chemistry},
volume = {59},
number = {3},
pages = {1140-1148},
abstract = {The dCTPase pyrophosphatase 1 (dCTPase) regulates the intracellular nucleotide pool through hydrolytic degradation of canonical and noncanonical nucleotide triphosphates (dNTPs). dCTPase is highly expressed in multiple carcinomas and is associated with cancer cell stemness. Here we report on the development of the first potent and selective dCTPase inhibitors that enhance the cytotoxic effect of cytidine analogues in leukemia cells. Boronate 30 displays a promising in vitro ADME profile, including plasma and mouse microsomal half-lives, aqueous solubility, cell permeability and CYP inhibition, deeming it a suitable compound for in vivo studies.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The dCTPase pyrophosphatase 1 (dCTPase) regulates the intracellular nucleotide pool through hydrolytic degradation of canonical and noncanonical nucleotide triphosphates (dNTPs). dCTPase is highly expressed in multiple carcinomas and is associated with cancer cell stemness. Here we report on the development of the first potent and selective dCTPase inhibitors that enhance the cytotoxic effect of cytidine analogues in leukemia cells. Boronate 30 displays a promising in vitro ADME profile, including plasma and mouse microsomal half-lives, aqueous solubility, cell permeability and CYP inhibition, deeming it a suitable compound for in vivo studies. |
Manara MC; Terracciano M; Mancarella C; Sciandra M; Guerzoni C; Pasello M; Grilli A; Zini N; Picci P; Colombo MP; Morrione A; Scotlandi K CD99 triggering induces methuosis of Ewing sarcoma cells through IGF-1R/RAS/Rac1 signaling. Journal Article In: Oncotarget, vol. 7, no 48, pp. 79925-79942, 2016. @article{%a1:%Y_295,
title = {CD99 triggering induces methuosis of Ewing sarcoma cells through IGF-1R/RAS/Rac1 signaling.},
author = {Manara MC and Terracciano M and Mancarella C and Sciandra M and Guerzoni C and Pasello M and Grilli A and Zini N and Picci P and Colombo MP and Morrione A and Scotlandi K},
url = {http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path[]=13160&pubmed-linkout=1},
doi = {10.18632/oncotarget.13160},
year = {2016},
date = {2016-02-18},
journal = {Oncotarget},
volume = {7},
number = {48},
pages = {79925-79942},
abstract = {CD99 is a cell surface molecule that has emerged as a novel target for Ewing sarcoma (EWS), an aggressive pediatric bone cancer. This report provides the first evidence of methuosis in EWS, a non-apoptotic form of cell death induced by an antibody directed against the CD99 molecule. Upon mAb triggering, CD99 induces an IGF-1R/RAS/Rac1 complex, which is internalized into RAB5-positive endocytic vacuoles. This complex is then dissociated, with the IGF-1R recycling to the cell membrane while CD99 and RAS/Rac1 are sorted into immature LAMP-1-positive vacuoles, whose excessive accumulation provokes methuosis. This process, which is not detected in CD99-expressing normal mesenchymal cells, is inhibited by disruption of the IGF-1R signaling, whereas enhanced by IGF-1 stimulation. Induction of IGF-1R/RAS/Rac1 was also observed in the EWS xenografts that respond to anti-CD99 mAb, further supporting the role of the IGF/RAS/Rac1 axis in the hyperstimulation of macropinocytosis and selective death of EWS cells. Thus, we describe a vulnerability of EWS cells, including those resistant to standard chemotherapy, to a treatment with anti-CD99 mAb, which requires IGF-1R/RAS signaling but bypasses the need for their direct targeting. Overall, we propose CD99 targeting as new opportunity to treat EWS patients resistant to canonical apoptosis-inducing agents.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
CD99 is a cell surface molecule that has emerged as a novel target for Ewing sarcoma (EWS), an aggressive pediatric bone cancer. This report provides the first evidence of methuosis in EWS, a non-apoptotic form of cell death induced by an antibody directed against the CD99 molecule. Upon mAb triggering, CD99 induces an IGF-1R/RAS/Rac1 complex, which is internalized into RAB5-positive endocytic vacuoles. This complex is then dissociated, with the IGF-1R recycling to the cell membrane while CD99 and RAS/Rac1 are sorted into immature LAMP-1-positive vacuoles, whose excessive accumulation provokes methuosis. This process, which is not detected in CD99-expressing normal mesenchymal cells, is inhibited by disruption of the IGF-1R signaling, whereas enhanced by IGF-1 stimulation. Induction of IGF-1R/RAS/Rac1 was also observed in the EWS xenografts that respond to anti-CD99 mAb, further supporting the role of the IGF/RAS/Rac1 axis in the hyperstimulation of macropinocytosis and selective death of EWS cells. Thus, we describe a vulnerability of EWS cells, including those resistant to standard chemotherapy, to a treatment with anti-CD99 mAb, which requires IGF-1R/RAS signaling but bypasses the need for their direct targeting. Overall, we propose CD99 targeting as new opportunity to treat EWS patients resistant to canonical apoptosis-inducing agents. |
Spoto G; De Iuliis V; Petrini M; Flati V; Di Gregorio J; Vitale D; Caruso M; Dadorante V; Ciarmoli M; Robuffo I; Martinotti S; Toniato E Effect of low energy light irradiation by light emitting diode on U937 cells. Journal Article In: Journal of Biological Regulators and Homeostatic Agents, vol. 30, no 4, pp. 997-1007, 2016. @article{%a1:%Y_312,
title = {Effect of low energy light irradiation by light emitting diode on U937 cells.},
author = {Spoto G and De Iuliis V and Petrini M and Flati V and Di Gregorio J and Vitale D and Caruso M and Dadorante V and Ciarmoli M and Robuffo I and Martinotti S and Toniato E},
url = {https://www.biolifesas.org/biolife/jbrha-2/},
year = {2016},
date = {2016-02-18},
journal = {Journal of Biological Regulators and Homeostatic Agents},
volume = {30},
number = {4},
pages = {997-1007},
abstract = {Photobiomodulation (PBM) can induce a set of different biological modulators either in vitro or in vivo. Experimental evidence has highlighted the role of light effects on the mechanisms related to inflammation, apoptosis and autophagy. The goal of this project was the evaluation of PBM on U937, an established cell line of histiocytic lymphoma origin. Several aspects of modulation of proinflammatory pathways were analyzed and autophagic and proapoptotic mechanisms related to low laser light exposure of cells were studied. As a source of low energy light emission, we used an NIR-LED device, characterized by an 880 nm-wavelength as light source. Flow cytometry analysis was performed on supernatants of controls and treated U937 cells to detect inflammatory cytokine levels. In order to evaluate NF-kB and caspase3 expressions, Western blot analysis was performed according to standard procedures. In this report, we show the effect of PBM on a monocyte/macrophage established tumor cell line (U-937). We demonstrate that LED exposure, in the presence or absence of lipopolysaccharide (LPS), activates cell degranulation, increased expression of Interleukin-8 (IL-8) and modulation of beta galactosidase activity. Evidence shows that the well-known pro-inflammatory nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) and the apoptotic marker (caspase3/cleaved-caspase3 ratio) are up-regulated in response to a proinflammatory biochemical pathway.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Photobiomodulation (PBM) can induce a set of different biological modulators either in vitro or in vivo. Experimental evidence has highlighted the role of light effects on the mechanisms related to inflammation, apoptosis and autophagy. The goal of this project was the evaluation of PBM on U937, an established cell line of histiocytic lymphoma origin. Several aspects of modulation of proinflammatory pathways were analyzed and autophagic and proapoptotic mechanisms related to low laser light exposure of cells were studied. As a source of low energy light emission, we used an NIR-LED device, characterized by an 880 nm-wavelength as light source. Flow cytometry analysis was performed on supernatants of controls and treated U937 cells to detect inflammatory cytokine levels. In order to evaluate NF-kB and caspase3 expressions, Western blot analysis was performed according to standard procedures. In this report, we show the effect of PBM on a monocyte/macrophage established tumor cell line (U-937). We demonstrate that LED exposure, in the presence or absence of lipopolysaccharide (LPS), activates cell degranulation, increased expression of Interleukin-8 (IL-8) and modulation of beta galactosidase activity. Evidence shows that the well-known pro-inflammatory nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) and the apoptotic marker (caspase3/cleaved-caspase3 ratio) are up-regulated in response to a proinflammatory biochemical pathway. |
Vasuvat J; Montree A; Moonsom S; Leartsakulpanich U; Petmitr S; Focher F; Wright GE; Chavalitshewinkoon-Petmitr P Biochemical and functional characterization of Plasmodium falciparum DNA polymerase delta. Journal Article In: Malaria Journal, vol. 15, no 1, pp. 116, 2016. @article{%a1:%Y_316,
title = {Biochemical and functional characterization of Plasmodium falciparum DNA polymerase delta.},
author = {Vasuvat J and Montree A and Moonsom S and Leartsakulpanich U and Petmitr S and Focher F and Wright GE and Chavalitshewinkoon-Petmitr P},
url = {http://malariajournal.biomedcentral.com/articles/10.1186/s12936-016-1166-0},
doi = {10.1186/s12936-016-1166-0},
year = {2016},
date = {2016-02-18},
journal = {Malaria Journal},
volume = {15},
number = {1},
pages = {116},
abstract = {BACKGROUND: Emergence of drug-resistant Plasmodium falciparum has created an urgent need for new drug targets. DNA polymerase delta is an essential enzyme required for chromosomal DNA replication and repair, and therefore may be a potential target for anti-malarial drug development. However, little is known of the characteristics and function of this P. falciparum enzyme. METHODS: The coding sequences of DNA polymerase delta catalytic subunit (PfPoldelta-cat), DNA polymerase delta small subunit (PfPoldeltaS) and proliferating cell nuclear antigen (PfPCNA) from chloroquine- and pyrimethamine-resistant P. falciparum strain K1 were amplified, cloned into an expression vector and expressed in Escherichia coli. The recombinant proteins were analysed by SDS-PAGE and identified by LC-MS/MS. PfPoldelta-cat was biochemically characterized. The roles of PfPoldeltaS and PfPCNA in PfPoldelta-cat function were investigated. In addition, inhibitory effects of 11 compounds were tested on PfPoldelta-cat activity and on in vitro parasite growth using SYBR Green I assay. RESULTS: The purified recombinant protein PfPoldelta-cat, PfPoldeltaS and PfPCNA showed on SDS-PAGE the expected size of 143, 57 and 34 kDa, respectively. Predicted amino acid sequence of the PfPoldelta-cat and PfPoldeltaS had 59.2 and 24.7 % similarity respectively to that of the human counterpart. The PfPoldelta-cat possessed both DNA polymerase and 3'-5' exonuclease activities. It used both Mg(2+) and Mn(2+) as cofactors and was inhibited by high KCl salt (>200 mM). PfPoldeltaS stimulated PfPoldelta-cat activity threefolds and up to fourfolds when PfPCNA was included in the assay. Only two compounds were potent inhibitors of PfPoldelta-cat, namely, butylphenyl-dGTP (BuPdGTP; IC50 of 38 µM) and 7-acetoxypentyl-(3, 4 dichlorobenzyl) guanine (7-acetoxypentyl-DCBG; IC50 of 55 µM). The latter compound showed higher inhibition on parasite growth (IC50 of 4.1 µM). CONCLUSIONS: Recombinant PfPoldelta-cat, PfPoldeltaS and PfPCNA were successfully expressed and purified. PfPolS and PfPCNA increased DNA polymerase activity of PfPoldelta-cat. The high sensitivity of PfPoldelta to BuPdGTP can be used to differentiate parasite enzyme from mammalian and human counterparts. Interestingly, 7-acetoxypentyl-DCBG showed inhibitory effects on both enzyme activity and parasite growth. Thus, 7-acetoxypentyl-DCBG is a potential candidate for future development of a new class of anti-malarial agents targeting parasite replicative DNA polymerase.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
BACKGROUND: Emergence of drug-resistant Plasmodium falciparum has created an urgent need for new drug targets. DNA polymerase delta is an essential enzyme required for chromosomal DNA replication and repair, and therefore may be a potential target for anti-malarial drug development. However, little is known of the characteristics and function of this P. falciparum enzyme. METHODS: The coding sequences of DNA polymerase delta catalytic subunit (PfPoldelta-cat), DNA polymerase delta small subunit (PfPoldeltaS) and proliferating cell nuclear antigen (PfPCNA) from chloroquine- and pyrimethamine-resistant P. falciparum strain K1 were amplified, cloned into an expression vector and expressed in Escherichia coli. The recombinant proteins were analysed by SDS-PAGE and identified by LC-MS/MS. PfPoldelta-cat was biochemically characterized. The roles of PfPoldeltaS and PfPCNA in PfPoldelta-cat function were investigated. In addition, inhibitory effects of 11 compounds were tested on PfPoldelta-cat activity and on in vitro parasite growth using SYBR Green I assay. RESULTS: The purified recombinant protein PfPoldelta-cat, PfPoldeltaS and PfPCNA showed on SDS-PAGE the expected size of 143, 57 and 34 kDa, respectively. Predicted amino acid sequence of the PfPoldelta-cat and PfPoldeltaS had 59.2 and 24.7 % similarity respectively to that of the human counterpart. The PfPoldelta-cat possessed both DNA polymerase and 3'-5' exonuclease activities. It used both Mg(2+) and Mn(2+) as cofactors and was inhibited by high KCl salt (>200 mM). PfPoldeltaS stimulated PfPoldelta-cat activity threefolds and up to fourfolds when PfPCNA was included in the assay. Only two compounds were potent inhibitors of PfPoldelta-cat, namely, butylphenyl-dGTP (BuPdGTP; IC50 of 38 µM) and 7-acetoxypentyl-(3, 4 dichlorobenzyl) guanine (7-acetoxypentyl-DCBG; IC50 of 55 µM). The latter compound showed higher inhibition on parasite growth (IC50 of 4.1 µM). CONCLUSIONS: Recombinant PfPoldelta-cat, PfPoldeltaS and PfPCNA were successfully expressed and purified. PfPolS and PfPCNA increased DNA polymerase activity of PfPoldelta-cat. The high sensitivity of PfPoldelta to BuPdGTP can be used to differentiate parasite enzyme from mammalian and human counterparts. Interestingly, 7-acetoxypentyl-DCBG showed inhibitory effects on both enzyme activity and parasite growth. Thus, 7-acetoxypentyl-DCBG is a potential candidate for future development of a new class of anti-malarial agents targeting parasite replicative DNA polymerase. |
Chiarini F; Lonetti A; Evangelisti C; Buontempo F; Orsini E; Evangelisti C; Cappellini A; Neri LM; McCubrey JA; Martelli AM Advances in understanding the acute lymphoblastic leukemia bone marrow microenvironment: From biology to therapeutic targeting. Journal Article In: Biochimica Et Biophysica Acta (BBA) - Molecular Cell Research, vol. 1863, no 3, pp. 449-463, 2016. @article{%a1:%Y_320,
title = {Advances in understanding the acute lymphoblastic leukemia bone marrow microenvironment: From biology to therapeutic targeting.},
author = {Chiarini F and Lonetti A and Evangelisti C and Buontempo F and Orsini E and Evangelisti C and Cappellini A and Neri LM and McCubrey JA and Martelli AM},
url = {http://www.sciencedirect.com/science/article/pii/S0167488915002931},
doi = {10.1016/j.bbamcr.2015.08.015. },
year = {2016},
date = {2016-02-18},
journal = {Biochimica Et Biophysica Acta (BBA) - Molecular Cell Research},
volume = {1863},
number = {3},
pages = {449-463},
abstract = {The bone marrow (BM) microenvironment regulates the properties of healthy hematopoietic stem cells (HSCs) localized in specific niches. Two distinct microenvironmental niches have been identified in the BM, the "osteoblastic (endosteal)" and "vascular" niches. Nevertheless, these niches provide sanctuaries where subsets of leukemic cells escape chemotherapy-induced death and acquire a drug-resistant phenotype. Moreover, it is emerging that leukemia cells are able to remodel the BM niches into malignant niches which better support neoplastic cell survival and proliferation. This review focuses on the cellular and molecular biology of microenvironment/leukemia interactions in acute lymphoblastic leukemia (ALL) of both B- and T-cell lineage. We shall also highlight the emerging role of exosomes/microvesicles as efficient messengers for cell-to-cell communication in leukemia settings. Studies on the interactions between the BM microenvironment and ALL cells have led to the discovery of potential therapeutic targets which include cytokines/chemokines and their receptors, adhesion molecules, signal transduction pathways, and hypoxia-related proteins. The complex interplays between leukemic cells and BM microenvironment components provide a rationale for innovative, molecularly targeted therapies, designed to improve ALL patient outcome. A better understanding of the contribution of the BM microenvironment to the process of leukemogenesis and leukemia persistence after initial remission, may provide new targets that will allow destruction of leukemia cells without adversely affecting healthy HSCs. This article is part of a Special Issue entitled: Tumor Microenvironment Regulation of Cancer Cell Survival, Metastasis,Inflammation, and Immune Surveillance. Guest Editors: Peter Ruvolo and Gregg L. Semenza. Copyright 2015 Elsevier B.V. All rights reserved.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The bone marrow (BM) microenvironment regulates the properties of healthy hematopoietic stem cells (HSCs) localized in specific niches. Two distinct microenvironmental niches have been identified in the BM, the "osteoblastic (endosteal)" and "vascular" niches. Nevertheless, these niches provide sanctuaries where subsets of leukemic cells escape chemotherapy-induced death and acquire a drug-resistant phenotype. Moreover, it is emerging that leukemia cells are able to remodel the BM niches into malignant niches which better support neoplastic cell survival and proliferation. This review focuses on the cellular and molecular biology of microenvironment/leukemia interactions in acute lymphoblastic leukemia (ALL) of both B- and T-cell lineage. We shall also highlight the emerging role of exosomes/microvesicles as efficient messengers for cell-to-cell communication in leukemia settings. Studies on the interactions between the BM microenvironment and ALL cells have led to the discovery of potential therapeutic targets which include cytokines/chemokines and their receptors, adhesion molecules, signal transduction pathways, and hypoxia-related proteins. The complex interplays between leukemic cells and BM microenvironment components provide a rationale for innovative, molecularly targeted therapies, designed to improve ALL patient outcome. A better understanding of the contribution of the BM microenvironment to the process of leukemogenesis and leukemia persistence after initial remission, may provide new targets that will allow destruction of leukemia cells without adversely affecting healthy HSCs. This article is part of a Special Issue entitled: Tumor Microenvironment Regulation of Cancer Cell Survival, Metastasis,Inflammation, and Immune Surveillance. Guest Editors: Peter Ruvolo and Gregg L. Semenza. Copyright 2015 Elsevier B.V. All rights reserved. |
Gelfo V; Rodia MT; Pucci M; Dall'Ora M; Santi S; Solmi R; Roth L; Lindzen M; Bonafè M; Bertotti A; Caramelli E; Lollini PL; Trusolino L; Yarden Y; D'Uva G; Lauriola M A module of inflammatory cytokines defines resistance of colorectal cancer to EGFR inhibitors. Journal Article In: Oncotarget, vol. 7, no 44, pp. 72167-72183, 2016. @article{%a1:%Y_282,
title = {A module of inflammatory cytokines defines resistance of colorectal cancer to EGFR inhibitors.},
author = {Gelfo V and Rodia MT and Pucci M and Dall'Ora M and Santi S and Solmi R and Roth L and Lindzen M and Bonafè M and Bertotti A and Caramelli E and Lollini PL and Trusolino L and Yarden Y and D'Uva G and Lauriola M},
url = {http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path[]=12354&pubmed-linkout=1},
doi = {10.18632/oncotarget.12354},
year = {2016},
date = {2016-02-17},
journal = {Oncotarget},
volume = {7},
number = {44},
pages = {72167-72183},
abstract = {Epidermal Growth Factor Receptor (EGFR) activates a robust signalling network to which colon cancer tumours often become addicted. Cetuximab, one of the monoclonal antibodies targeting this pathway, is employed to treat patients with colorectal cancer. However, many patients are intrinsically refractory to this treatment, and those who respond develop secondary resistance along time. Mechanisms of cancer cell resistance include either acquisition of new mutations or non genomic activation of alternative signalling routes. In this study, we employed a colon cancer model to assess potential mechanisms driving resistance to cetuximab. Resistant cells displayed increased ability to grow in suspension as colonspheres and this phenotype was associated with poorly organized structures. Factors secreted from resistant cells were causally involved in sustaining resistance, indeed administration to parental cells of conditioned medium collected from resistant cells was sufficient to reduce cetuximab efficacy. Among secreted factors, we report herein that a signature of inflammatory cytokines, including IL1A, IL1B and IL8, which are produced following EGFR pathway activation, was associated with the acquisition of an unresponsive phenotype to cetuximab in vitro. This signature correlated with lack of response to EGFR targeting also in patient-derived tumour xenografts. Collectively, these results highlight the contribution of inflammatory cytokines to reduced sensitivity to EGFR blockade and suggest that inhibition of this panel of cytokines in combination with cetuximab might yield an effective treatment strategy for CRC patients refractory to anti-EGFR targeting.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Epidermal Growth Factor Receptor (EGFR) activates a robust signalling network to which colon cancer tumours often become addicted. Cetuximab, one of the monoclonal antibodies targeting this pathway, is employed to treat patients with colorectal cancer. However, many patients are intrinsically refractory to this treatment, and those who respond develop secondary resistance along time. Mechanisms of cancer cell resistance include either acquisition of new mutations or non genomic activation of alternative signalling routes. In this study, we employed a colon cancer model to assess potential mechanisms driving resistance to cetuximab. Resistant cells displayed increased ability to grow in suspension as colonspheres and this phenotype was associated with poorly organized structures. Factors secreted from resistant cells were causally involved in sustaining resistance, indeed administration to parental cells of conditioned medium collected from resistant cells was sufficient to reduce cetuximab efficacy. Among secreted factors, we report herein that a signature of inflammatory cytokines, including IL1A, IL1B and IL8, which are produced following EGFR pathway activation, was associated with the acquisition of an unresponsive phenotype to cetuximab in vitro. This signature correlated with lack of response to EGFR targeting also in patient-derived tumour xenografts. Collectively, these results highlight the contribution of inflammatory cytokines to reduced sensitivity to EGFR blockade and suggest that inhibition of this panel of cytokines in combination with cetuximab might yield an effective treatment strategy for CRC patients refractory to anti-EGFR targeting. |
Radi M; Schneider R; Fallacara AL; Botta L; Crespan E; Tintori C; Maga G; Kissova M; Calgani A; Richters A; Musumeci F; Rauh D; Schenone S A cascade screening approach for the identification of Bcr-Abl myristate pocket binders active against wild type and T315I mutant. Journal Article In: Bioorganic & Medicinal Chemistry Letters, vol. 26, no 15, pp. 3436-3440, 2016. @article{%a1:%Y_304,
title = {A cascade screening approach for the identification of Bcr-Abl myristate pocket binders active against wild type and T315I mutant.},
author = {Radi M and Schneider R and Fallacara AL and Botta L and Crespan E and Tintori C and Maga G and Kissova M and Calgani A and Richters A and Musumeci F and Rauh D and Schenone S},
url = {http://www.sciencedirect.com/science/article/pii/S0960894X1630662X},
doi = {10.1016/j.bmcl.2016.06.051},
year = {2016},
date = {2016-02-17},
journal = {Bioorganic & Medicinal Chemistry Letters},
volume = {26},
number = {15},
pages = {3436-3440},
abstract = {The major clinical challenge in drug-resistant chronic myelogenous leukemia (CML) is currently represented by the Bcr-Abl T315I mutant, which is unresponsive to treatment with common first and second generation ATP-competitive tyrosine kinase inhibitors (TKIs). Allosteric inhibition of Bcr-Abl represent a new frontier in the fight against resistant leukemia and few candidates have been identified in the last few years. Among these, myristate pocket (MP) binders discovered by Novartis (e.g. GNF2/5) showed promising results, although they proved to be active against the T315I mutant only in combination with first and second generation ATP-competitive inhibitors. Here we used a cascade screening approach based on sequential fluorescence polarization (FP) screening, in silico docking/dynamics studies and kinetic-enzymatic studies to identify novel MP binders. A pyrazolo[3,4-d]pyrimidine derivative (6) has been identified as a promising allosteric inhibitor active on 32D leukemia cell lines (expressing Bcr-Abl WT and T315I) with no need of combination with any ATP-competitive inhibitor.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The major clinical challenge in drug-resistant chronic myelogenous leukemia (CML) is currently represented by the Bcr-Abl T315I mutant, which is unresponsive to treatment with common first and second generation ATP-competitive tyrosine kinase inhibitors (TKIs). Allosteric inhibition of Bcr-Abl represent a new frontier in the fight against resistant leukemia and few candidates have been identified in the last few years. Among these, myristate pocket (MP) binders discovered by Novartis (e.g. GNF2/5) showed promising results, although they proved to be active against the T315I mutant only in combination with first and second generation ATP-competitive inhibitors. Here we used a cascade screening approach based on sequential fluorescence polarization (FP) screening, in silico docking/dynamics studies and kinetic-enzymatic studies to identify novel MP binders. A pyrazolo[3,4-d]pyrimidine derivative (6) has been identified as a promising allosteric inhibitor active on 32D leukemia cell lines (expressing Bcr-Abl WT and T315I) with no need of combination with any ATP-competitive inhibitor. |
Sardone F; Santi S; Tagliavini F; Traina F; Merlini L; Squarzoni S; Cescon M; Wagener R; Maraldi NM; Bonaldo P; Faldini C; Sabatelli P Collagen VI-NG2 axis in human tendon fibroblasts under conditions mimicking injury response. Journal Article In: Matrix Biology, vol. 55, pp. 90-105, 2016. @article{%a1:%Y_307,
title = {Collagen VI-NG2 axis in human tendon fibroblasts under conditions mimicking injury response.},
author = {Sardone F and Santi S and Tagliavini F and Traina F and Merlini L and Squarzoni S and Cescon M and Wagener R and Maraldi NM and Bonaldo P and Faldini C and Sabatelli P},
url = {https://www.sciencedirect.com/science/article/pii/S0945053X16300233?via%3Dihub},
doi = {10.1016/j.matbio.2016.02.012},
year = {2016},
date = {2016-02-17},
journal = {Matrix Biology},
volume = {55},
pages = {90-105},
abstract = {In response to injury, tendon fibroblasts are activated, migrate to the wound, and contribute to tissue repair by producing and organizing the extracellular matrix. Collagen VI is a microfibrillar collagen enriched in the pericellular matrix of tendon fibroblasts with a potential regulatory role in tendon repair mechanism. We investigated the molecular basis of the interaction between collagen VI and the cell membrane both in tissue sections and fibroblast cultures of human tendon, and analyzed the deposition of collagen VI during migration and myofibroblast trans-differentiation, two crucial events for tendon repair. Tendon fibroblast displayed a collagen VI microfibrillar network closely associated with the cell surface. Binding of collagen VI with the cell membrane was mediated by NG2 proteoglycan, as demonstrated by in vitro perturbation of collagen VI-NG2 interaction with a NG2-blocking antibody. Cultures subjected to wound healing scratch assay displayed collagen VI-NG2 complexes at the trailing edge of migrating cells, suggesting a potential role in cell migration. In fact, the addition of a NG2-blocking antibody led to an impairment of cell polarization and delay of wound closure. Similar results were obtained after in vitro perturbation of collagen VI extracellular assembly with the 3C4 anti-collagen VI antibody and in collagen VI-deficient tendon cultures of a Ullrich congenital muscular dystrophy patient carrying mutations in COL6A2 gene. Moreover, in vitro treatment with transforming growth factor beta1 (TGFbeta1) induced a dramatic reduction of NG2 expression, both at protein and mRNA transcript level, and the impairment of collagen VI association with the cell membrane. Instead, collagen VI was still detectable in the extracellular matrix in association with ED-A fibronectin and collagen I, which were strongly induced by TGFbeta1 treatment. Our findings reveal a critical role of the NG2 proteoglycan for the binding of collagen VI to the surface of tendon fibroblasts. By interacting with NG2 proteoglycan and other extracellular matrix proteins, collagen VI regulates fibroblasts behavior and the assembly of tendon matrix, thereby playing a crucial role in tendon repair. Copyright 2016 The Authors. Published by Elsevier B.V. All rights reserved.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
In response to injury, tendon fibroblasts are activated, migrate to the wound, and contribute to tissue repair by producing and organizing the extracellular matrix. Collagen VI is a microfibrillar collagen enriched in the pericellular matrix of tendon fibroblasts with a potential regulatory role in tendon repair mechanism. We investigated the molecular basis of the interaction between collagen VI and the cell membrane both in tissue sections and fibroblast cultures of human tendon, and analyzed the deposition of collagen VI during migration and myofibroblast trans-differentiation, two crucial events for tendon repair. Tendon fibroblast displayed a collagen VI microfibrillar network closely associated with the cell surface. Binding of collagen VI with the cell membrane was mediated by NG2 proteoglycan, as demonstrated by in vitro perturbation of collagen VI-NG2 interaction with a NG2-blocking antibody. Cultures subjected to wound healing scratch assay displayed collagen VI-NG2 complexes at the trailing edge of migrating cells, suggesting a potential role in cell migration. In fact, the addition of a NG2-blocking antibody led to an impairment of cell polarization and delay of wound closure. Similar results were obtained after in vitro perturbation of collagen VI extracellular assembly with the 3C4 anti-collagen VI antibody and in collagen VI-deficient tendon cultures of a Ullrich congenital muscular dystrophy patient carrying mutations in COL6A2 gene. Moreover, in vitro treatment with transforming growth factor beta1 (TGFbeta1) induced a dramatic reduction of NG2 expression, both at protein and mRNA transcript level, and the impairment of collagen VI association with the cell membrane. Instead, collagen VI was still detectable in the extracellular matrix in association with ED-A fibronectin and collagen I, which were strongly induced by TGFbeta1 treatment. Our findings reveal a critical role of the NG2 proteoglycan for the binding of collagen VI to the surface of tendon fibroblasts. By interacting with NG2 proteoglycan and other extracellular matrix proteins, collagen VI regulates fibroblasts behavior and the assembly of tendon matrix, thereby playing a crucial role in tendon repair. Copyright 2016 The Authors. Published by Elsevier B.V. All rights reserved. |
Scotton C; Bovolenta M; Schwartz E; Falzarano MS; Martoni E; Passarelli C; Armaroli A; Osman H; Rodolico C; Messina S; Pegoraro E; D'Amico A; Bertini E; Gualandi F; Neri M; Selvatici R; Boffi P; Maioli MA; Lochmüller H; Straub V; Bushby K; Castrignanò T; Pesole G; Sabatelli P; Merlini L; Braghetta P; Bonaldo P; Bernardi P; Foley R; Cirak S; Zaharieva I; Muntoni F; Capitanio D; Gelfi C; Kotelnikova E; Yuryev A; Lebowitz M; Zhang X; Hodge B; Esser KA; Ferlini A Deep RNA profiling identified clock and molecular clock genes as pathophysiological signatures in collagen VI myopathy. Journal Article In: Journal of Cell Science, vol. 129, no 8, pp. 1671-1684, 2016. @article{%a1:%Y_310,
title = {Deep RNA profiling identified clock and molecular clock genes as pathophysiological signatures in collagen VI myopathy.},
author = {Scotton C and Bovolenta M and Schwartz E and Falzarano MS and Martoni E and Passarelli C and Armaroli A and Osman H and Rodolico C and Messina S and Pegoraro E and D'Amico A and Bertini E and Gualandi F and Neri M and Selvatici R and Boffi P and Maioli MA and Lochmüller H and Straub V and Bushby K and Castrignanò T and Pesole G and Sabatelli P and Merlini L and Braghetta P and Bonaldo P and Bernardi P and Foley R and Cirak S and Zaharieva I and Muntoni F and Capitanio D and Gelfi C and Kotelnikova E and Yuryev A and Lebowitz M and Zhang X and Hodge B and Esser KA and Ferlini A},
url = {http://jcs.biologists.org/content/early/2016/03/04/jcs.175927.long},
doi = {10.1242/jcs.175927},
year = {2016},
date = {2016-02-17},
journal = {Journal of Cell Science},
volume = {129},
number = {8},
pages = {1671-1684},
abstract = {Collagen VI myopathies are genetic disorders due to mutations in collagen 6 A1, 2, and 3 genes, ranging from the severe Ullrich congenital muscular dystrophy to the milder Bethlem Myopathy, which is recapitulated by collagen VI null (Col6a1-/-) mice. Abnormalities in mitochondria and autophagic pathway have been proposed as pathogenic causes of collagen VI myopathies, but the link between collagen VI defects and these metabolic circuits remains unknown. To unravel the expression profiling perturbation in muscles with collagen VI myopathies we performed a deep RNA profiling in both Col6a1-/- mice and ColVI patients. Interactome map identified common pathways suggesting a previously undetected connection between circadian genes and collagen VI pathology. Intriguingly, Bmal1-/- mice, a well-characterized model displaying arrhythmic circadian rhythms, showed profound deregulation of the collagen VI pathway and autophagy-related genes.The involvement of circadian rhythms in collagen VI myopathies is new and links autophagy and mitochondrial abnormalities. It also opens new avenues for therapies of hereditary myopathies to modulate the molecular clock or potential gene-environment interactions that may modify muscle damage pathogenesis. 2016. Published by The Company of Biologists Ltd.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Collagen VI myopathies are genetic disorders due to mutations in collagen 6 A1, 2, and 3 genes, ranging from the severe Ullrich congenital muscular dystrophy to the milder Bethlem Myopathy, which is recapitulated by collagen VI null (Col6a1-/-) mice. Abnormalities in mitochondria and autophagic pathway have been proposed as pathogenic causes of collagen VI myopathies, but the link between collagen VI defects and these metabolic circuits remains unknown. To unravel the expression profiling perturbation in muscles with collagen VI myopathies we performed a deep RNA profiling in both Col6a1-/- mice and ColVI patients. Interactome map identified common pathways suggesting a previously undetected connection between circadian genes and collagen VI pathology. Intriguingly, Bmal1-/- mice, a well-characterized model displaying arrhythmic circadian rhythms, showed profound deregulation of the collagen VI pathway and autophagy-related genes.The involvement of circadian rhythms in collagen VI myopathies is new and links autophagy and mitochondrial abnormalities. It also opens new avenues for therapies of hereditary myopathies to modulate the molecular clock or potential gene-environment interactions that may modify muscle damage pathogenesis. 2016. Published by The Company of Biologists Ltd. |
Zlatanou A; Sabbioneda S; Miller ES; Greenwalt A; Aggathanggelou A; Maurice MM; Lehmann AR; Stankovic T; Reverdy C; Colland F; Vaziri C; Stewart GS USP7 is essential for maintaining Rad18 stability and DNA damage tolerance. Journal Article In: Oncogene, vol. 35, no 8, pp. 965-976, 2016. @article{%a1:%Y_319,
title = {USP7 is essential for maintaining Rad18 stability and DNA damage tolerance.},
author = {Zlatanou A and Sabbioneda S and Miller ES and Greenwalt A and Aggathanggelou A and Maurice MM and Lehmann AR and Stankovic T and Reverdy C and Colland F and Vaziri C and Stewart GS},
url = {http://www.nature.com/onc/journal/vaop/ncurrent/full/onc2015149a.html},
doi = {10.1038/onc.2015.149},
year = {2016},
date = {2016-02-16},
journal = {Oncogene},
volume = {35},
number = {8},
pages = {965-976},
abstract = {965 976},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
|
Avnet S; Lemma S; Cortini M; Pellegrini P; Perut F; Zini N; Kusuzaki K; Chano T; Grisendi G; Dominici M; De Milito A; Baldini N Altered pH gradient at the plasma membrane of osteosarcoma cells is a key mechanism of drug resistance. Journal Article In: Oncotarget, vol. 7, no 39, pp. 63408-63423, 2016. @article{%a1:%Y_248,
title = {Altered pH gradient at the plasma membrane of osteosarcoma cells is a key mechanism of drug resistance.},
author = {Avnet S and Lemma S and Cortini M and Pellegrini P and Perut F and Zini N and Kusuzaki K and Chano T and Grisendi G and Dominici M and De Milito A and Baldini N},
url = {http://www.impactjournals.com/oncotarget/index.php?journal=oncotarget&page=article&op=view&path[]=11503&pubmed-linkout=1},
doi = {10.18632/oncotarget.11503},
year = {2016},
date = {2016-02-12},
journal = {Oncotarget},
volume = {7},
number = {39},
pages = {63408-63423},
abstract = {Current therapy of osteosarcoma (OS), the most common primary bone malignancy, is based on a combination of surgery and chemotherapy. Multidrug resistance mediated by P-glycoprotein (P-gp) overexpression has been previously associated with treatment failure and progression of OS, although other mechanisms may also play a role. We considered the typical acidic extracellular pH (pHe) of sarcomas, and found that doxorubicin (DXR) cytotoxicity is reduced in P-gp negative OS cells cultured at pHe 6.5 compared to standard 7.4. Short-time (24-48 hours) exposure to low pHe significantly increased the number and acidity of lysosomes, and the combination of DXR with omeprazole, a proton pump inhibitor targeting lysosomal acidity, significantly enhanced DXR cytotoxicity. In OS xenografts, the combination treatment of DXR and omeprazole significantly reduced tumor volume and body weight loss. The impaired toxicity of DXR at low pHe was not associated with increased autophagy or lysosomal acidification, but rather, as shown by SNARF staining, with a reversal of the pH gradient at the plasma membrane (ΔpHcm), eventually leading to a reduced DXR intracellular accumulation. Finally, the reversal of ΔpHcm in OS cells promoted resistance not only to DXR, but also to cisplatin and methotrexate, and, to a lesser extent, to vincristine. Altogether, our findings show that, in OS cells, short-term acidosis induces resistance to different chemotherapeutic drugs by a reversal of ΔpHcm, suggesting that buffer therapies or regimens including proton pump inhibitors in combination to low concentrations of conventional anticancer agents may offer novel solutions to overcome drug resistance.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Current therapy of osteosarcoma (OS), the most common primary bone malignancy, is based on a combination of surgery and chemotherapy. Multidrug resistance mediated by P-glycoprotein (P-gp) overexpression has been previously associated with treatment failure and progression of OS, although other mechanisms may also play a role. We considered the typical acidic extracellular pH (pHe) of sarcomas, and found that doxorubicin (DXR) cytotoxicity is reduced in P-gp negative OS cells cultured at pHe 6.5 compared to standard 7.4. Short-time (24-48 hours) exposure to low pHe significantly increased the number and acidity of lysosomes, and the combination of DXR with omeprazole, a proton pump inhibitor targeting lysosomal acidity, significantly enhanced DXR cytotoxicity. In OS xenografts, the combination treatment of DXR and omeprazole significantly reduced tumor volume and body weight loss. The impaired toxicity of DXR at low pHe was not associated with increased autophagy or lysosomal acidification, but rather, as shown by SNARF staining, with a reversal of the pH gradient at the plasma membrane (ΔpHcm), eventually leading to a reduced DXR intracellular accumulation. Finally, the reversal of ΔpHcm in OS cells promoted resistance not only to DXR, but also to cisplatin and methotrexate, and, to a lesser extent, to vincristine. Altogether, our findings show that, in OS cells, short-term acidosis induces resistance to different chemotherapeutic drugs by a reversal of ΔpHcm, suggesting that buffer therapies or regimens including proton pump inhibitors in combination to low concentrations of conventional anticancer agents may offer novel solutions to overcome drug resistance. |
Croce AC; Ferrigno A; Di Pasqua LG; Berardo C; Piccolini VM; Bertone V; Bottiroli G; Vairetti M Autofluorescence discrimination of metabolic fingerprint in nutritional and genetic fatty liver models. Journal Article In: Journal of Photochemistry and Photobiology. B, Biology., vol. 164, pp. 13-20, 2016. @article{%a1:%Y_264,
title = {Autofluorescence discrimination of metabolic fingerprint in nutritional and genetic fatty liver models.},
author = {Croce AC and Ferrigno A and Di Pasqua LG and Berardo C and Piccolini VM and Bertone V and Bottiroli G and Vairetti M},
url = {http://www.sciencedirect.com/science/article/pii/S1011134416302913},
doi = {10.1016/j.jphotobiol.2016.09.015},
year = {2016},
date = {2016-02-11},
journal = {Journal of Photochemistry and Photobiology. B, Biology.},
volume = {164},
pages = {13-20},
abstract = {Liver tissue autofluorescence (AF) has been characterized in two models with a different potential to undergo disease progression to steatohepatitis: Wistar rats, administered with a methionine, choline deficient diet (MCD), and Zucker (fa/fa) rats, homozygous for a spontaneous mutation of leptin receptor. AF spectra were recorded from liver tissue cryostatic sections by microspectrofluorometry, under 366nm excitation. Curve fitting analysis was used to estimate the contribution of different endogenous fluorophores (EFs) to the overall AF emission: i) fluorescing fatty acids, a fraction of liver lipids up to now poorly considered and complicated to detect by conventional procedures; ii) lipofuscin-like lipopigments, biomarkers of oxidizing events; iii) NAD(P)H and flavins, biomarkers of energy metabolism and tissue redox state. AF data and biochemical correlates of hepatocellular injury resulted to depend more on rat strain than on intratissue bulk lipid or ROS levels, reflecting a different metabolic ability of the two models to counteract potentially harmful agents. AF analysis can thus be proposed for extensive applications ranging from experimental hepatology to the clinics. AF based diagnostic procedures are expected to help both the prediction of the risk of fatty liver disease progression and the prescreening of marginal organs to be recruited as donors for transplantation. A support is also foreseen in the advancement and personalization of strategies to ameliorate the donor organ preservation outcome and the follow up of therapeutic interventions. Copyright 2016 Elsevier B.V. All rights reserved.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Liver tissue autofluorescence (AF) has been characterized in two models with a different potential to undergo disease progression to steatohepatitis: Wistar rats, administered with a methionine, choline deficient diet (MCD), and Zucker (fa/fa) rats, homozygous for a spontaneous mutation of leptin receptor. AF spectra were recorded from liver tissue cryostatic sections by microspectrofluorometry, under 366nm excitation. Curve fitting analysis was used to estimate the contribution of different endogenous fluorophores (EFs) to the overall AF emission: i) fluorescing fatty acids, a fraction of liver lipids up to now poorly considered and complicated to detect by conventional procedures; ii) lipofuscin-like lipopigments, biomarkers of oxidizing events; iii) NAD(P)H and flavins, biomarkers of energy metabolism and tissue redox state. AF data and biochemical correlates of hepatocellular injury resulted to depend more on rat strain than on intratissue bulk lipid or ROS levels, reflecting a different metabolic ability of the two models to counteract potentially harmful agents. AF analysis can thus be proposed for extensive applications ranging from experimental hepatology to the clinics. AF based diagnostic procedures are expected to help both the prediction of the risk of fatty liver disease progression and the prescreening of marginal organs to be recruited as donors for transplantation. A support is also foreseen in the advancement and personalization of strategies to ameliorate the donor organ preservation outcome and the follow up of therapeutic interventions. Copyright 2016 Elsevier B.V. All rights reserved. |
Ettorrea V; De Marcoa P; Zaraa S; Perrottib V; Scarano A; Di Crescenzo A; Petrini M; Hadad C; Bosco D; Zavan B; Valbonetti L; Spoto G; Iezzi G; Piattelli A; Cataldi A; Fontana A In vitro and in vivo characterization of graphene oxide coated porcine bone granules Journal Article In: Carbon, vol. 103, pp. 291-298, 2016. @article{%a1:%Y_273,
title = {In vitro and in vivo characterization of graphene oxide coated porcine bone granules},
author = {Ettorrea V and De Marcoa P and Zaraa S and Perrottib V and Scarano A and Di Crescenzo A and Petrini M and Hadad C and Bosco D and Zavan B and Valbonetti L and Spoto G and Iezzi G and Piattelli A and Cataldi A and Fontana A},
url = {https://www.sciencedirect.com/science/article/pii/S0008622316301993},
doi = {10.1016/j.carbon.2016.03.010},
year = {2016},
date = {2016-02-11},
journal = {Carbon},
volume = {103},
pages = {291-298},
abstract = {Graphene oxide (GO) demonstrated to improve the wound healing properties of materials intended for bone replacement. The main objective of this study was the setting up of a simple and effective procedure for the production of GO-coated porcine bone (PB) granules and the characterization of the obtained material in order to improve its properties by exploiting chemical, physical, biological and mechanical features that the GO coating could confer to pre-formed PB granules. The obtained coating was homogeneously distributed on PB granule surface and demonstrated to confer PB an increased resistance to fracture load. Biological analyses evidenced no toxic effects of GO-coated PB samples on primary human gingival fibroblasts, and no inflammatory response around the grafted particles when implanted in vivo on a sheep model although GO-coated PB samples did not appear to improve new bone formation efficacy compared with the control within the investigated time. A small loss of GO was however detected, indicating the opportunity to investigate less GO concentrated samples. In conclusion, this study presents a novel and low cost approach to the development of functionalized biomimetic hybrid materials which can be applied to other bone substitute materials in order to improve their performances.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Graphene oxide (GO) demonstrated to improve the wound healing properties of materials intended for bone replacement. The main objective of this study was the setting up of a simple and effective procedure for the production of GO-coated porcine bone (PB) granules and the characterization of the obtained material in order to improve its properties by exploiting chemical, physical, biological and mechanical features that the GO coating could confer to pre-formed PB granules. The obtained coating was homogeneously distributed on PB granule surface and demonstrated to confer PB an increased resistance to fracture load. Biological analyses evidenced no toxic effects of GO-coated PB samples on primary human gingival fibroblasts, and no inflammatory response around the grafted particles when implanted in vivo on a sheep model although GO-coated PB samples did not appear to improve new bone formation efficacy compared with the control within the investigated time. A small loss of GO was however detected, indicating the opportunity to investigate less GO concentrated samples. In conclusion, this study presents a novel and low cost approach to the development of functionalized biomimetic hybrid materials which can be applied to other bone substitute materials in order to improve their performances. |
Fan Q; Guo X; Tideman JW; Williams KM; Yazar S; Hosseini SM; Howe LD; Pourcain BS; Evans DM; Timpson NJ; McMahon G; Hysi PG; Krapohl E; Wang YX; Jonas JB; Baird PN; Wang JJ; Cheng CY; Teo YY; Wong TY; Ding X; Wojciechowski R; Young TL; Pärssinen O; Oexle K; Pfeiffer N; Bailey-Wilson JE; Paterson AD; Klaver CC; Plomin R; Hammond CJ; Mackey DA; He M; Saw SM; Williams C; Guggenheim JA; CREAM Consortium Childhood gene-environment interactions and age-dependent effects of genetic variants associated with refractive error and myopia: The CREAM Consortium. Journal Article In: Scientific Reports, vol. 6, pp. 25853, 2016. @article{%a1:%Y_276,
title = {Childhood gene-environment interactions and age-dependent effects of genetic variants associated with refractive error and myopia: The CREAM Consortium.},
author = {Fan Q and Guo X and Tideman JW and Williams KM and Yazar S and Hosseini SM and Howe LD and Pourcain BS and Evans DM and Timpson NJ and McMahon G and Hysi PG and Krapohl E and Wang YX and Jonas JB and Baird PN and Wang JJ and Cheng CY and Teo YY and Wong TY and Ding X and Wojciechowski R and Young TL and Pärssinen O and Oexle K and Pfeiffer N and Bailey-Wilson JE and Paterson AD and Klaver CC and Plomin R and Hammond CJ and Mackey DA and He M and Saw SM and Williams C and Guggenheim JA and CREAM Consortium},
url = {http://www.nature.com/articles/srep25853},
doi = {10.1038/srep25853},
year = {2016},
date = {2016-02-11},
journal = {Scientific Reports},
volume = {6},
pages = {25853},
abstract = {Myopia, currently at epidemic levels in East Asia, is a leading cause of untreatable visual impairment. Genome-wide association studies (GWAS) in adults have identified 39 loci associated with refractive error and myopia. Here, the age-of-onset of association between genetic variants at these 39 loci and refractive error was investigated in 5200 children assessed longitudinally across ages 7-15 years, along with gene-environment interactions involving the major environmental risk-factors, nearwork and time outdoors. Specific variants could be categorized as showing evidence of: (a) early-onset effects remaining stable through childhood, (b) early-onset effects that progressed further with increasing age, or (c) onset later in childhood (N = 10, 5 and 11 variants, respectively). A genetic risk score (GRS) for all 39 variants explained 0.6% (P = 6.6E-08) and 2.3% (P = 6.9E-21) of the variance in refractive error at ages 7 and 15, respectively, supporting increased effects from these genetic variants at older ages. Replication in multi-ancestry samples (combined N = 5599) yielded evidence of childhood onset for 6 of 12 variants present in both Asians and Europeans. There was no indication that variant or GRS effects altered depending on time outdoors, however 5 variants showed nominal evidence of interactions with nearwork (top variant, rs7829127 in ZMAT4; P = 6.3E-04).},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Myopia, currently at epidemic levels in East Asia, is a leading cause of untreatable visual impairment. Genome-wide association studies (GWAS) in adults have identified 39 loci associated with refractive error and myopia. Here, the age-of-onset of association between genetic variants at these 39 loci and refractive error was investigated in 5200 children assessed longitudinally across ages 7-15 years, along with gene-environment interactions involving the major environmental risk-factors, nearwork and time outdoors. Specific variants could be categorized as showing evidence of: (a) early-onset effects remaining stable through childhood, (b) early-onset effects that progressed further with increasing age, or (c) onset later in childhood (N = 10, 5 and 11 variants, respectively). A genetic risk score (GRS) for all 39 variants explained 0.6% (P = 6.6E-08) and 2.3% (P = 6.9E-21) of the variance in refractive error at ages 7 and 15, respectively, supporting increased effects from these genetic variants at older ages. Replication in multi-ancestry samples (combined N = 5599) yielded evidence of childhood onset for 6 of 12 variants present in both Asians and Europeans. There was no indication that variant or GRS effects altered depending on time outdoors, however 5 variants showed nominal evidence of interactions with nearwork (top variant, rs7829127 in ZMAT4; P = 6.3E-04). |