Maurizio Mauro - Academia.edu (original) (raw)

Papers by Maurizio Mauro

Nutrients

Autophagy is an evolutionarily conserved process critical in maintaining cellular homeostasis. Re... more Autophagy is an evolutionarily conserved process critical in maintaining cellular homeostasis. Recently, the anticancer potential of autophagy inducers, including phytochemicals, was suggested. Indicaxanthin is a betalain pigment found in prickly pear fruit with antiproliferative and pro-apoptotic activities in colorectal cancer cells associated with epigenetic changes in selected methylation-silenced oncosuppressor genes. Here, we demonstrate that indicaxanthin induces the up-regulation of the autophagic markers LC3-II and Beclin1, and increases autophagolysosome production in Caco-2 cells. Methylomic studies showed that the indicaxanthin-induced pro-autophagic activity was associated with epigenetic changes. In addition to acting as a hypermethylating agent at the genomic level, indicaxanthin also induced significant differential methylation in 39 out of 47 autophagy-related genes, particularly those involved in the late stages of autophagy. Furthermore, in silico molecular modell...

Atti XI Congresso SIGU, 2008

Le pi\uf9 recenti stime dell\u2019OMS testimoniano che le patologie correlate all\u2019abitudine ... more Le pi\uf9 recenti stime dell\u2019OMS testimoniano che le patologie correlate all\u2019abitudine al fumo e al consumo di alcool sono tra le principali cause di morte nei soggetti adulti ed \ue8 noto che la diversit\ue0 interindividuale nell\u2019adottare tali stili di vita \ue8 attribuibile, oltre che a fattori psico-sociali, anche a polimorfismi dei geni per i citocromi P450. Questo studio mira a verificare se esista una correlazione fra i genotipi CYP2A6 -CYP2E1 e abitudine al fumo e consumo d\u2019alcool nella Sicilia centro-occidentale. I risultati ottenuti dall\u2019analisi della distribuzione di alcuni alleli di entrambi i geni in 65 soggetti (41 donne e 24 uomini) hanno evidenziato che i genotipi CYP2A6*1/*4A e CYP2A6*1/*4B sono prevalenti in soggetti non fumatori (28.8% vs 5.1% nei fumatori) e il genotipo omozigote CYP2E1*C1/*C1 \ue8 prevalente rispetto a quello eterozigote CYP2E1*C1/*C2 in soggetti che assumono bevande alcoliche (26.32% vs 19.22%). Sorprendentemente, nel 33,8% dei casi \ue8 stato riscontrato un genotipo CYP2E1 eterozigote per il sito di restrizione RsaI ed omozigote per il sito di restrizione PstI, condizione non attesa visto il linkage disequilibrium descritto per questi due siti. Inoltre, per entrambi i geni, \ue8 stata rilevata una differente distribuzione genotipica in relazione al sesso. Infine, l\u2019associazione genotipica CYP2A6*1/*4B-CYP2E1*C1/*C1, che conferisce bassa suscettibilit\ue0 nei confronti di patologie neoplastiche fumo-alcool correlate, \ue8 risultata essere la pi\uf9 frequente (41.51%)

Cancer Research, 2010

ABSTRACT Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC Abstract F... more ABSTRACT Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC Abstract Fanconi anemia (FA) is a rare recessive disorder characterized in part by pronounced cancer susceptibility. The FA proteins, and the protein products of the major breast cancer susceptibility genes BRCA1 and BRCA2, function cooperatively in the FA-BRCA pathway, to repair damaged DNA and to prevent cellular transformation. A major step in the activation of the FA-BRCA pathway is the mono-ubiquitination of the FANCD2 and FANCI proteins, targeting these proteins to discrete chromatin-associated nuclear foci. Importantly, the regulation of FANCD2 (and FANCI) mono-ubiquitination remains poorly understood. To gain further insight into this important post-translational modification step, here we have examined the roles of p53 tumor suppressor as well its downstream target p21Cip1/Waf1 in the regulation of the mono-ubiquitination of FANCD2. In this study we have used the isogenic HCT116 p53+/+, p53−/−, p21+/+, and p21−/− cell lines, to determine that p21, and not p53, plays a key role in the regulation of the mono-ubiquitination of FANCD2 following exposure to DNA damaging agents. For example, robust FANCD2 mono-ubiquitination is observed in HCT116 p21+/+ cells 2 h following exposure to 20 J/m2 UV irradiation, while this effect is markedly attenuated in p21−/− cells. Using immunofluorescence microscopy we also demonstrate that p21 is necessary for the efficient recruitment of FANCD2 to nuclear foci following exposure to UV irradiation. Furthermore, preliminary protein-protein interaction analyses in COS-7 cells demonstrate that FLAG-p21 and 6xHis/V5-FANCD2 physically interact. Interestingly, despite a failure to efficiently activate the mono-ubiquitination of FANCD2 following exposure to DNA damaging agents, p21−/− cells are not hypersensitive to the cytotoxic effects of the DNA crosslinking agent mitomycin C (MMC). However, elevated levels of distinct chromosome aberrations, including telomere associations and dicentric chromosomes, are observed in p21−/− cells compared with p21+/+ cells following exposure to MMC. Taken together, our results demonstrate an important upstream role for the p21 cyclin-dependent-kinase inhibitor in the activation of the FA-BRCA pathway. A greater understanding of the regulation of this important tumor suppressor pathway will lead to improved diagnostic and therapeutic approaches to FA and enhance our understanding of the elevated cancer susceptibility of FA patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-102. ©2010 American Association for Cancer Research

From Genes to …, 2006

... Citazione: SCIANDRELLO G, MAURO M, CATANZARO I, CARADONNA F, & BARBATA G. (2006). Persist... more ... Citazione: SCIANDRELLO G, MAURO M, CATANZARO I, CARADONNA F, & BARBATA G. (2006). Persistent genomic instability by arsenic exposure in V79 Chinese hamster cells.. ... Titolo: Persistent genomic instability by arsenic exposure in V79 Chinese hamster cells. ...

… on line degli atti del IV …, 2006

... damaging DNA by utilizing Comet assay, which showed a dose-dependent increase of tail lenght,... more ... damaging DNA by utilizing Comet assay, which showed a dose-dependent increase of tail lenght, in metabolically non competent V79 Chinese hamster cells. Moreover, we observed that Acrylamide (AA) was able to antagonize in vivo the citotoxicity of well ...

Nucleic Acids Research, 2012

p21 is a well-established regulator of cell cycle progression. The role of p21 in DNA repair, how... more p21 is a well-established regulator of cell cycle progression. The role of p21 in DNA repair, however, remains poorly characterized. Here, we describe a critical role of p21 in a replication-coupled DNA double-strand break (DSB) repair that is mechanistically distinct from its cell cycle checkpoint function. We demonstrate that p21-deficient cells exhibit elevated chromatid-type aberrations, including gaps and breaks, dicentrics and radial formations, following exposure to several DSBinducing agents. p21 À/À cells also exhibit an increased DNA damage-inducible DNA-PK CS S2056 phosphorylation, indicative of elevated nonhomologous DNA end joining. Concomitantly, p21 À/À cells are defective in replication-coupled homologous recombination (HR), exhibiting decreased sister chromatid exchanges and HRdependent repair as determined using a crosslinked GFP reporter assay. Importantly, we establish that the DSB hypersensitivity of p21 À/À cells is associated with increased cyclin-dependent kinase (CDK)-dependent BRCA2 S3291 phosphorylation and MRE11 nuclear foci formation and can be rescued by inhibition of CDK or MRE11 nuclease activity. Collectively, our results uncover a novel mechanism by which p21 regulates the fidelity of replication-coupled DSB repair and the maintenance of chromosome stability distinct from its role in the G1-S phase checkpoint.

Oncotarget, 2017

Fanconi anemia (FA) is a rare disease characterized by congenital defects, bone marrow failure, a... more Fanconi anemia (FA) is a rare disease characterized by congenital defects, bone marrow failure, and atypically early-onset cancers. The FA proteins function cooperatively to repair DNA interstrand crosslinks. A major step in the activation of the pathway is the monoubiquitination of the FANCD2 and FANCI proteins, and their recruitment to chromatin-associated nuclear foci. The regulation and function of FANCD2 and FANCI, however, is poorly understood. In addition, how chromatin state impacts pathway activation is also unknown. In this study, we have examined the influence of chromatin state on the activation of the FA pathway. We describe potent activation of FANCD2 and FANCI monoubiquitination and nuclear foci formation following treatment of cells with the histone methyltransferase inhibitor BRD4770. BRD4770-induced activation of the pathway does not occur via the direct induction of DNA damage or via the inhibition of the G9a histone methyltransferase, a mechanism previously proposed for this molecule. Instead, we show that BRD4770inducible FANCD2 and FANCI monoubiquitination and nuclear foci formation may be a consequence of inhibition of the PRC2/EZH2 chromatin-modifying complex. In addition, we show that inhibition of the class I and II histone deacetylases leads to attenuated FANCD2 and FANCI monoubiquitination and nuclear foci formation. Our studies establish that chromatin state is a major determinant of the activation of the FA pathway and suggest an important role for the PRC2/EZH2 complex in the regulation of this critical tumor suppressor pathway.

Mitosis is regulated by checkpoints that delay mitotic progression when chromosome segregation er... more Mitosis is regulated by checkpoints that delay mitotic progression when chromosome segregation errors occur. Inaccuracy in checkpoint processes can lead to chromosome instability both in number and structure (CIN). Arsenic is reported to induce CIN by perturbing mitotic spindles and checkpoints, however, its carcinogenic mechanisms are poorly understood. We previously studied the long-term progression of chromosomal instability in V79 cells treated acutely with arsenite (10 \uf06dM, 24 hr) followed by growth in arsenic-free medium for 120 cell generations, and found time-dependent increase of aneuploid cells. Here, we treated V79-derived G12 cells with sub-lethal doses (0.1-1.0 \ub5M) of arsenite for 10 days followed by growth in arsenite-free medium for 40 cell generations. Cytogenetic analysis at the end of treatment showed concentration-dependent increases in the frequency of aneuploid cells that was even higher after 40 cell generations. Furthermore, the mitotic index (MI) of arsenite-exposed cells was higher than untreated cells at the end of the 10-day treatment or after 40 cell generations without arsenite. Surprisingly, we found that submicromolar 10-day arsenic exposure induced a large amount of cells in anaphase with premature centromere division. These aberrant mitotic figures dramatically increased after 40 cell generations. Taken together, these results raise the possibility that chronic exposure to sub-lethal arsenic doses induces bypass of the spindle assembly checkpoints and may be mechanistically involved in the induction of cell transformation

Previously we demonstrated that V79 Chinese hamster cells underwent either early genetic instabil... more Previously we demonstrated that V79 Chinese hamster cells underwent either early genetic instability or apoptosis When exposed to sodium arsenite (SA). Genetic instability was evidenced by aneuploid and morphologically abnormal cells, but not by cells with chromosome aberrations. As dividing cells turned out to be the most sensitive to SA exposure, due to the arsenics direct action on the mitotic spindle assembly, we later ascertained the fate of genetically unstable cells escaping apoptosis, by harvesting mitotic rounded-up cells at the end of a 24 h treatment. The progeny of the exposed Chinese hamster cells showed an increased level of mutations related to genome DNA hypomethylation induced by arsenic after treatment. In fact, cytogenetic, morphological and molecular investigations, performed at several time points during the expanded growth in drug-free medium, emphasized that genomic instability reappeared since 60th cell generation. Metaphases with dicentric chromosomes or telomeric associations characterized cell population since 90th cell generation. Some of the isolated clones also bearing chromosomal end-to-end fusions maintained telomerase activity and were capable to proliferate accumulating genomic instability as well as transformed phenotype and spontaneous increased gene mutation oxidative stress associated. On the whole, these results raise the possibility that the short-term exposure to arsenic induces altered DNA methylation pattern conferring a selective advantage to cell variants. Similarly to Karpinets and Foy hypothesis (Carcinogenesis, 2005) we believe these unstable cells epigenetically reprogram their genome and proliferate in an error-prone mod

The thermal heat conductivity, specific heat capacity, thermal heat diffusivity and bulk density ... more The thermal heat conductivity, specific heat capacity, thermal heat diffusivity and bulk density of Prosopis africana seeds were determined as a function of moisture content. Specific heat capacity was measured by the method of mixture while the thermal heat conductivity was measured by the guarded hot plate method. Thermal heat diffusivity was calculated from the experimental results obtained from specific heat capacity, thermal heat conductivity and bulk density. The bulk density for Prosopis africana (PA) seeds decreased from 890kg m-3 to 590kg m-3 as moisture content increased from 4 to 20% wet basis (w.b). Specific heat capacity increased from 2760J kg-1 ºC-1 to 2960J kg-1 ºC-1 with increasing moisture content. The thermal heat conductivity ranged between 0.70 and 0.90W m-1o C-1 when moisture content rose from 4% to 20% (w.b). Thermal heat diffusivity increased from 2.7  10-7 to 4.2  10-7 m 2 s-1 as moisture content increased from 4 to 20% (w.b). The values obtained for these thermal properties and bulk density could be useful for design of systems for heat treatment of Prosopis africana seeds.

Fertility and Sterility, 2019

To study the effects of insulin and metformin on primary trophoblasts from early pregnancies. Des... more To study the effects of insulin and metformin on primary trophoblasts from early pregnancies. Design: Experimental in vitro study. Setting: Academic research institute. Patient(s): Trophoblasts from healthy patients undergoing first trimester elective termination of pregnancy and primary lung fibroblasts (IMR-90). Intervention(s): Culture and treatment with insulin and metformin of primary trophoblasts and primary lung fibroblasts (IMR-90). Main Outcome Measure(s): DNA damage measured by expression of g-H2AX with immunofluorescence and Western blot. Apoptosis measured by expression of cleaved caspase-3 by Western blot. Cell survival measured by cell proliferation assay. Result(s): Culture of purified primary trophoblast cells in the presence of insulin at levels as low as 1 nM resulted in a 386% increase in the number of cell with elevated g-H2AX expression, a 66% reduction in cell survival and a marked increase of cleaved caspase-3 expression. Pretreatment of trophoblasts with therapeutic doses of metformin prevented the detrimental effects of insulin. Treatment with insulin and/or metformin had no effects on primary fibroblasts. Conclusion(s): Elevated insulin levels are directly toxic to first trimester trophoblasts and result in increased DNA damage, apoptosis, and decreased cell survival. These effects are prevented by metformin. Trophoblast cells from early pregnancy are uniquely vulnerable to elevated levels of insulin. These findings, if confirmed in vivo, suggest that there may be a role for insulin resistance screening before attempting pregnancy and for focusing on prevention of hyperinsulinemia during early pregnancy. (Fertil Steril Ò 2019;111:489-96. Ó2018 by American Society for Reproductive Medicine.) El resumen está disponible en Español al final del artículo.

Mutagenesis, 2016

Arsenic (AS) is a reactive oxygen species (ROS)-inducer carcinogen, whose mode of action is still... more Arsenic (AS) is a reactive oxygen species (ROS)-inducer carcinogen, whose mode of action is still unclear. To defend against ROS, cells use enzymatic and non-enzymatic antioxidants, such as superoxide dismutase (SOD) and catalase. Failure of antioxidant systems (AXS) can result in dicentric chromosomes formation as well as telomere associations for the reduced activity of telomerase. In order to clarify the long-term effects of a past AS exposure, we evaluated the efficiency of the AXS and the telomerase activity in the progeny of arsenite-treated cells named ASO (arsenic shake-off) cells, previously obtained from arsenite-treated V79 cells and selected by shake-off. Despite SOD 1 expression level correlated to the level of ROS observed over time, no changes of the relative amount of antioxidant activities were observed in ASO cells. Moreover, we found that clones characterised by low levels of SOD 1 and high levels of ROS acquired a transformed phenotype. Treatment with 5-azacytidine determined an increase of SOD 1 expression in a clone and decrease in one other, suggesting that aberrant DNA methylation may be responsible for the abnormal expression of SOD1 or SOD 1 inhibitor genes in different clones. TRAP assay results showed that the progeny of arsenite-treated cells were characterised by a time-dependent decrease of telomerase activity. Integrated results suggest that the increases of ROS levels are accompanied by defective telomerase activity. Finally, we propose that cells escaping the arseniteinduced death perpetuated the memory of past exposure via ROS likely because antioxidant and telomerase activity impairment and ultimately acquire a transformed phenotype.

Environmental and molecular mutagenesis, Jan 19, 2015

The mechanisms by which arsenic-induced genomic instability is initiated and maintained are poorl... more The mechanisms by which arsenic-induced genomic instability is initiated and maintained are poorly understood. To investigate potential epigenetic mechanisms, in this study we evaluated global DNA methylation levels in V79 cells and human HaCaT keratinocytes at several time points during expanded growth of cell cultures following removal of arsenite exposures. We have found altered genomic methylation patterns that persisted up to 40 cell generations in HaCaT cells after the treatments were withdrawn. Moreover, mRNA expression levels were evaluated by RT-PCR for DNMT1, DNMT3A, DNMT3B, HMLH1, and HMSH2 genes, demonstrating that the down regulation of DNMT3A and DNMT3B genes, but not DNMT1, occurred in an arsenic dose-dependent manner, and persisted for many cell generations following removal of the arsenite, offering a plausible mechanism of persistently genotoxic arsenic action. Analyses of promoter methylation status of the DNA mismatch repair genes HMLH1 and HMSH2 show that HMSH2,...

BioTechniques, 2015

Extracellular RNAs (exRNAs) are present in biofluids such as breast milk, cerebrospinal fluid, pl... more Extracellular RNAs (exRNAs) are present in biofluids such as breast milk, cerebrospinal fluid, plasma, saliva, and urine, and comprise a wide spectrum of RNA species, including microRNAs (miRNAs), messenger RNAs (mRNAs), ribosomal RNAs (rRNAs), transfer RNAs (tRNAs), small nuclear RNAs (snRNAs), and long non-coding RNAs (lncRNAs) (1). Successful detection of organ-specific and disease-associated exRNAs from plasma or serum samples by PCR, microarray and, more recently, RNA-seq has positioned circulating exRNAs as promising candidates for non-invasive biomarker discovery (2-6). However,

BMC Molecular Biology, 2015

Background: RNA quantification is often a prerequisite for most RNA analyses such as RNA sequenci... more Background: RNA quantification is often a prerequisite for most RNA analyses such as RNA sequencing. However, the relatively low sensitivity and large sample consumption of traditional RNA quantification methods such as UV spectrophotometry and even the much more sensitive fluorescence-based RNA quantification assays, such as the Qubit™ RNA HS Assay, are often inadequate for measuring minute levels of RNA isolated from limited cell and tissue samples and biofluids. Thus, there is a pressing need for a more sensitive method to reliably and robustly detect trace levels of RNA without interference from DNA. Methods: To improve the quantification limit of the Qubit™ RNA HS Assay, we spiked-in a known quantity of RNA to achieve the minimum reading required by the assay. Samples containing trace amounts of RNA were then added to the spike-in and measured as a reading increase over RNA spike-in baseline. We determined the accuracy and precision of reading increases between 1 and 20 pg/μL as well as RNA-specificity in this range, and compared to those of RiboGreen®, another sensitive fluorescence-based RNA quantification assay. We then applied Qubit™ Assay with RNA spike-in to quantify plasma RNA samples. Results: RNA spike-in improved the quantification limit of the Qubit™ RNA HS Assay 5-fold, from 25 pg/μL down to 5 pg/μL while maintaining high specificity to RNA. This enabled quantification of RNA with original concentration as low as 55.6 pg/μL compared to 250 pg/μL for the standard assay and decreased sample consumption from 5 to 1 ng. Plasma RNA samples that were not measurable by the Qubit™ RNA HS Assay were measurable by our modified method. Conclusions: The Qubit™ RNA HS Assay with RNA spike-in is able to quantify RNA with high specificity at 5-fold lower concentration and uses 5-fold less sample quantity than the standard Qubit™ Assay.

Mutagenesis, 2012

Here, we report the effects of exposure of mammalian cells to a-pinene, a bicyclic monoterpene us... more Here, we report the effects of exposure of mammalian cells to a-pinene, a bicyclic monoterpene used in insecticides, solvents and perfumes. Morphological analysis, performed in V79-Cl3 cells exposed for 1 h to increasing concentrations (25 up to 50 mM) of a-pinene, indicated a statistically significant increase in micronucleated and multinucleated cell frequencies; apoptotic cells were seen at 40 and 50 mM. This monoterpene caused genomic instability by interfering with mitotic process; in fact, 50% of cells (versus 19% of control cells) showed irregular mitosis with multipolar or incorrectly localised spindles. Cytogenetic analysis demonstrated high-frequency hypodiploid metaphases as well as endoreduplicated cells and chromosome breaks. Clastogenic damage was prevalent over aneuploidogenic damage as demonstred by the higher proportion of kinetochore-negative micronuclei. Alkaline comet confirmed that monoterpene exposure caused DNA lesions in a concentration-dependent manner. This damage probably arose by increased reactive oxygen species (ROS) production. In order to assess the generation of ROS, the cells were incubated with CM-H 2 DCFDA and then analysed by flow cytometry. Results demonstrated an increase in fluorescence intensity after a-pinene treatment indicating increased oxidative stress. On the whole, these findings strongly suggest that a-pinene is able to compromise genome stability preferentially through mitotic alterations and to damage DNA through ROS production.

Toxicology in Vitro, 2010

The vinyl monomer acrylamide is characterized by the presence of an a,b-unsaturated carbonyl grou... more The vinyl monomer acrylamide is characterized by the presence of an a,b-unsaturated carbonyl group that makes it reactive towards thiol, hydroxyl or amino groups and towards the nucleophilic centers in DNA. The ability of acrylamide to chemically modify protein thiols has prompted us to consider topoisomerase II as one possible target of acrylamide, since agents targeting protein sulfhydryl groups act as either catalytic inhibitors or poisons of topoisomerase II. Nuclear extracts from V79 Chinese hamster cells incubated with acrylamide reduced topoisomerase II activity as inferred by an inability to convert kinetoplast DNA to the decatenated form. Nuclear extracts incubated with acrylamide pre-incubated with DTT converted kinetoplast DNA to the decatenated form, suggesting that acrylamide influences topoisomerase II activity through reaction with sulfhydryl groups on the enzyme. Furthermore, acrylamide did not induce the pBR322 DNA cleavage, as assessed by cleavage assay; thus, it cannot be regarded as a poison of topoisomerase II. As a catalytic inhibitor, acrylamide antagonizes the effect of etoposide, a topoisomerase II poison, as determined by clonogenic assay in V79 cells. This antagonism is confirmed by band depletion assay, from which it can be inferred that acrylamide reduces the level of catalytically active cellular topoisomerase II available for the action of etoposide.