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1.
Oncogene ; 40(6): 1077-1090, 2021 02.
Article in English | MEDLINE | ID: mdl-33323971

ABSTRACT

The mutagenic APOBEC3B (A3B) cytosine deaminase is frequently over-expressed in cancer and promotes tumour heterogeneity and therapy resistance. Hence, understanding the mechanisms that underlie A3B over-expression is important, especially for developing therapeutic approaches to reducing A3B levels, and consequently limiting cancer mutagenesis. We previously demonstrated that A3B is repressed by p53 and p53 mutation increases A3B expression. Here, we investigate A3B expression upon treatment with chemotherapeutic drugs that activate p53, including 5-fluorouracil, etoposide and cisplatin. Contrary to expectation, these drugs induced A3B expression and concomitant cellular cytosine deaminase activity. A3B induction was p53-independent, as chemotherapy drugs stimulated A3B expression in p53 mutant cells. These drugs commonly activate ATM, ATR and DNA-PKcs. Using specific inhibitors and gene knockdowns, we show that activation of DNA-PKcs and ATM by chemotherapeutic drugs promotes NF-κB activity, with consequent recruitment of NF-κB to the A3B gene promoter to drive A3B expression. Further, we find that A3B knockdown re-sensitises resistant cells to cisplatin, and A3B knockout enhances sensitivity to chemotherapy drugs. Our data highlight a role for A3B in resistance to chemotherapy and indicate that stimulation of A3B expression by activation of DNA repair and NF-κB pathways could promote cancer mutations and expedite chemoresistance.


Subject(s)
Cytidine Deaminase/genetics , Minor Histocompatibility Antigens/genetics , Neoplasms/genetics , Transcription Factor RelA/genetics , Tumor Suppressor Protein p53/genetics , Ataxia Telangiectasia Mutated Proteins/genetics , CRISPR-Cas Systems/genetics , Cisplatin/pharmacology , DNA Repair/drug effects , Etoposide/pharmacology , Fluorouracil/pharmacology , Gene Expression Regulation, Neoplastic/drug effects , Genetic Heterogeneity , HCT116 Cells , Humans , MCF-7 Cells , Mutation/genetics , NF-kappa B/genetics , Neoplasms/pathology
2.
Elife ; 92020 09 21.
Article in English | MEDLINE | ID: mdl-32956035

ABSTRACT

Numerous anti-cancer drugs perturb thymidylate biosynthesis and lead to genomic uracil incorporation contributing to their antiproliferative effect. Still, it is not yet characterized if uracil incorporations have any positional preference. Here, we aimed to uncover genome-wide alterations in uracil pattern upon drug treatments in human cancer cell line models derived from HCT116. We developed a straightforward U-DNA sequencing method (U-DNA-Seq) that was combined with in situ super-resolution imaging. Using a novel robust analysis pipeline, we found broad regions with elevated probability of uracil occurrence both in treated and non-treated cells. Correlation with chromatin markers and other genomic features shows that non-treated cells possess uracil in the late replicating constitutive heterochromatic regions, while drug treatment induced a shift of incorporated uracil towards segments that are normally more active/functional. Data were corroborated by colocalization studies via dSTORM microscopy. This approach can be applied to study the dynamic spatio-temporal nature of genomic uracil.


Subject(s)
Antineoplastic Agents/pharmacology , DNA , Genome , Uracil , DNA/analysis , DNA/biosynthesis , DNA/chemistry , DNA/genetics , Genome/drug effects , Genome/genetics , Genomics , HCT116 Cells , Humans , Microscopy , Sequence Analysis, DNA , Uracil/analysis , Uracil/biosynthesis , Uracil/chemistry
3.
IEEE Trans Haptics ; 13(1): 233-238, 2020.
Article in English | MEDLINE | ID: mdl-32012026

ABSTRACT

In the previous work, we reported that tactile information (tactile slip) during finger sliding and reaching actions over a textured surface contributes to the control of the hand movement. More specifically, we observed a significant bias in the motion trajectories, which was explained by the tactile estimate accounted by the tactile flow model-i.e. a perceived motion direction always perpendicular to the ridge orientation, and its integration with the muscular-skeletal proprioceptive cues. However, to which extent this observed behavior also depends on the reaction force exerted by the surface ridges on the finger pad during the dynamic interaction still represents a largely unexplored research question. If not properly addressed, this point could rise the alternative explanation that the systematic bias is determined by the insufficient compensation of the reaction force by participants. In this article, we investigate the role of the lateral component of the reaction force on the surface plane (lateral force) in texture-related motion bias. We asked participants to slide their finger straight on a lubricated ridged plate towards a target goal displayed in a virtual reality environment. They exerted two different levels of normal force, which produced two different levels of lateral force during the finger interaction with the ridges. The effect of ridge orientation was found to be larger for the high compared to the low force level. However, also in the latter case, we still observed the same biased trajectories reported in our previous work, despite the negligible value of the lateral force. This supports our hypothesis that the motor bias arises from the integration of the tactile motion estimate, biased by the texture, and the other proprioceptive cues.


Subject(s)
Motion Perception , Psychomotor Performance , Touch Perception , Adult , Biomechanical Phenomena , Female , Fingers , Humans , Male , Proprioception , Surface Properties , Virtual Reality
4.
IEEE Trans Haptics ; 13(1): 239-245, 2020.
Article in English | MEDLINE | ID: mdl-32012027

ABSTRACT

Blindness represents one of the major disabling societal causes, impacting the life of visually impaired people and their families. For what concerns the access to written information, one of the main tools used by blind people is the traditional Braille code. This is the reason why in the recent years, there has been a technological effort to develop refreshable Braille devices. These consist of multiple physical dots that dynamically change their configuration to reproduce different sequences of the letters in Braille code. Although promising, these approaches have many drawbacks, which are mainly related to costs, design complexity, portability, and power consumption. Of note, while many solutions have been proposed for multi-cell devices, the investigation of the potentialities of single-cell refreshable systems has received little attention so far. This investigation could offer effective and viable manners to overcome the aforementioned drawbacks, likely fostering a widespread adoption of such assistive technologies with end-users. In this article, we present the design and characterization of a new cost-effective single-cell Electromagnetic Refreshable Braille Display, the Readable system. We also report on tests performed with blindfolded and blind expert Braille code readers. Results demonstrate the effectiveness of our device in correctly reproducing alphanumeric content, opening promising perspectives in every-day life applications.


Subject(s)
Blindness , Communication Aids for Disabled , Reading , Sensory Aids , Touch Perception , Touch , Adult , Electromagnetic Phenomena , Equipment Design , Female , Humans , Male , Middle Aged , User-Computer Interface
5.
Oncogene ; 39(12): 2523-2538, 2020 03.
Article in English | MEDLINE | ID: mdl-31988455

ABSTRACT

Glioblastoma (GBM) is the most common and most aggressive intrinsic brain tumour in adults. Integrated transcriptomic and epigenomic analyses of glioblastoma initiating cells (GIC) in a mouse model uncovered a novel epigenetic regulation of EfnA5. In this model, Bmi1 enhances H3K27me3 at the EfnA5 locus and reinforces repression of selected target genes in a cellular context-dependent fashion. EfnA5 mediates Bmi1-dependent proliferation and invasion in vitro and tumour formation in an allograft model. Importantly, we show that this novel Polycomb feed-forward loop is also active in human GIC and we provide pre-clinical evidence of druggability of the EFNA5 signalling pathway in GBM xenografts overexpressing Bmi1.


Subject(s)
Ephrin-A5/metabolism , Glioblastoma/metabolism , Polycomb Repressive Complex 1/metabolism , Animals , Antihypertensive Agents/pharmacology , Cell Proliferation , Doxazosin/pharmacology , Drug Delivery Systems , Ephrin-A5/antagonists & inhibitors , Epigenesis, Genetic , Gene Expression Regulation, Neoplastic , Glioblastoma/pathology , Histones/metabolism , Humans , Lysine/metabolism , Mice , Mice, Transgenic , Neoplasm Invasiveness , Neural Stem Cells/metabolism , Neurogenesis , Polycomb Repressive Complex 1/genetics , Tumor Cells, Cultured
6.
Elife ; 72018 10 26.
Article in English | MEDLINE | ID: mdl-30362940

ABSTRACT

An essential step for understanding the transcriptional circuits that control development and physiology is the global identification and characterization of regulatory elements. Here, we present the first map of regulatory elements across the development and ageing of an animal, identifying 42,245 elements accessible in at least one Caenorhabditis elegans stage. Based on nuclear transcription profiles, we define 15,714 protein-coding promoters and 19,231 putative enhancers, and find that both types of element can drive orientation-independent transcription. Additionally, more than 1000 promoters produce transcripts antisense to protein coding genes, suggesting involvement in a widespread regulatory mechanism. We find that the accessibility of most elements changes during development and/or ageing and that patterns of accessibility change are linked to specific developmental or physiological processes. The map and characterization of regulatory elements across C. elegans life provides a platform for understanding how transcription controls development and ageing.


Subject(s)
Aging/metabolism , Caenorhabditis elegans/growth & development , Caenorhabditis elegans/metabolism , Chromatin/metabolism , Animals , Caenorhabditis elegans/genetics , DNA/genetics , Enhancer Elements, Genetic , Gene Expression Regulation, Developmental , Histone Code , Histones/metabolism , Molecular Sequence Annotation , Promoter Regions, Genetic , Reproducibility of Results , Transcription Factors/metabolism , Transcription Initiation Site
7.
Nucleic Acids Res ; 45(19): 11056-11069, 2017 Nov 02.
Article in English | MEDLINE | ID: mdl-28977491

ABSTRACT

Cancer genome sequencing has implicated the cytosine deaminase activity of apolipoprotein B mRNA editing enzyme catalytic polypeptide-like (APOBEC) genes as an important source of mutations in diverse cancers, with APOBEC3B (A3B) expression especially correlated with such cancer mutations. To better understand the processes directing A3B over-expression in cancer, and possible therapeutic avenues for targeting A3B, we have investigated the regulation of A3B gene expression. Here, we show that A3B expression is inversely related to p53 status in different cancer types and demonstrate that this is due to a direct and pivotal role for p53 in repressing A3B expression. This occurs through the induction of p21 (CDKN1A) and the recruitment of the repressive DREAM complex to the A3B gene promoter, such that loss of p53 through mutation, or human papilloma virus-mediated inhibition, prevents recruitment of the complex, thereby causing elevated A3B expression and cytosine deaminase activity in cancer cells. As p53 is frequently mutated in cancer, our findings provide a mechanism by which p53 loss can promote cancer mutagenesis.


Subject(s)
Cytidine Deaminase/genetics , Gene Expression Regulation, Neoplastic , Minor Histocompatibility Antigens/genetics , Tumor Suppressor Protein p53/genetics , Cell Line , Cyclin-Dependent Kinase Inhibitor p21/genetics , Cyclin-Dependent Kinase Inhibitor p21/metabolism , Cytidine Deaminase/metabolism , HCT116 Cells , Humans , Immunoblotting , Minor Histocompatibility Antigens/metabolism , Mutation , Neoplasms/genetics , Neoplasms/metabolism , Neoplasms/pathology , RNA Interference , Reverse Transcriptase Polymerase Chain Reaction , Tumor Suppressor Protein p53/metabolism
8.
Science ; 353(6298): 495-8, 2016 Jul 29.
Article in English | MEDLINE | ID: mdl-27386920

ABSTRACT

A suboptimal early-life environment, due to poor nutrition or stress during pregnancy, can influence lifelong phenotypes in the progeny. Epigenetic factors are thought to be key mediators of these effects. We show that protein restriction in mice from conception until weaning induces a linear correlation between growth restriction and DNA methylation at ribosomal DNA (rDNA). This epigenetic response remains into adulthood and is restricted to rDNA copies associated with a specific genetic variant within the promoter. Related effects are also found in models of maternal high-fat or obesogenic diets. Our work identifies environmentally induced epigenetic dynamics that are dependent on underlying genetic variation and establishes rDNA as a genomic target of nutritional insults.


Subject(s)
DNA, Ribosomal/genetics , Epigenesis, Genetic , Gene-Environment Interaction , Maternal Nutritional Physiological Phenomena , Nutritional Status , Animals , DNA Methylation , Diet, High-Fat , Diet, Protein-Restricted , Female , Genetic Variation , Male , Mice , Obesity/genetics , Pregnancy , Promoter Regions, Genetic , Weaning
9.
Clin Cancer Res ; 22(23): 5929-5938, 2016 Dec 01.
Article in English | MEDLINE | ID: mdl-27301701

ABSTRACT

PURPOSE: CDK-activating kinase (CAK) is required for the regulation of the cell cycle and is a trimeric complex consisting of cyclin-dependent kinase 7 (CDK7), Cyclin H, and the accessory protein, MAT1. CDK7 also plays a critical role in regulating transcription, primarily by phosphorylating RNA polymerase II, as well as transcription factors such as estrogen receptor-α (ER). Deregulation of cell cycle and transcriptional control are general features of tumor cells, highlighting the potential for the use of CDK7 inhibitors as novel cancer therapeutics. EXPERIMENTAL DESIGN: mRNA and protein expression of CDK7 and its essential cofactors cyclin H and MAT1 were evaluated in breast cancer samples to determine if their levels are altered in cancer. Immunohistochemical staining of >900 breast cancers was used to determine the association with clinicopathologic features and patient outcome. RESULTS: We show that expressions of CDK7, cyclin H, and MAT1 are all closely linked at the mRNA and protein level, and their expression is elevated in breast cancer compared with the normal breast tissue. Intriguingly, CDK7 expression was inversely proportional to tumor grade and size, and outcome analysis showed an association between CAK levels and better outcome. Moreover, CDK7 expression was positively associated with ER expression and in particular with phosphorylation of ER at serine 118, a site important for ER transcriptional activity. CONCLUSIONS: Expressions of components of the CAK complex, CDK7, MAT1, and Cyclin H are elevated in breast cancer and correlate with ER. Like ER, CDK7 expression is inversely proportional to poor prognostic factors and survival. Clin Cancer Res; 22(23); 5929-38. ©2016 AACR.


Subject(s)
Breast Neoplasms/genetics , Breast Neoplasms/pathology , Carrier Proteins/genetics , Cyclin H/genetics , Cyclin-Dependent Kinases/genetics , Gene Expression/genetics , Receptors, Estrogen/genetics , Adult , Cell Cycle Proteins , Female , Humans , Middle Aged , Phosphorylation/genetics , Prognosis , Signal Transduction/genetics , Transcription Factors , Transcription, Genetic/genetics , Cyclin-Dependent Kinase-Activating Kinase
10.
BMC Genomics ; 16: 295, 2015 Apr 14.
Article in English | MEDLINE | ID: mdl-25888192

ABSTRACT

BACKGROUND: As sex determines mammalian development, understanding the nature and developmental dynamics of the sexually dimorphic transcriptome is important. To explore this, we generated 76 genome-wide RNA-seq profiles from mouse eight-cell embryos, late gestation and adult livers, together with 4 ground-state pluripotent embryonic (ES) cell lines from which we generated both RNA-seq and multiple ChIP-seq profiles. We complemented this with previously published data to yield 5 snap-shots of pre-implantation development, late-gestation placenta and somatic tissue and multiple adult tissues for integrative analysis. RESULTS: We define a high-confidence sex-dimorphic signature of 69 genes in eight-cell embryos. Sex-chromosome-linked components of this signature are largely conserved throughout pre-implantation development and in ES cells, whilst the autosomal component is more dynamic. Sex-biased gene expression is reflected by enrichment for activating and repressive histone modifications. The eight-cell signature is largely non-overlapping with that defined from fetal liver, neither was it correlated with adult liver or other tissues analysed. The number of sex-dimorphic genes increases throughout development. We identified many more dimorphic genes in adult compared to fetal liver. However, approximately two thirds of the dimorphic genes identified in fetal liver were also dimorphic in adult liver. Sex-biased expression differences unique to adult liver were enriched for growth hormone-responsiveness. Sexually dimorphic gene expression in pre-implantation development is driven by sex-chromosome based transcription, whilst later development is characterised by sex dimorphic autosomal transcription. CONCLUSION: This systematic study identifies three distinct phases of sex dimorphism throughout mouse development, and has significant implications for understanding the developmental origins of sex-specific phenotypes and disease in mammals.


Subject(s)
Embryonic Development/genetics , Genome , Sex Characteristics , Transcriptome/genetics , Animals , Embryo, Mammalian , Female , Gene Expression Regulation, Developmental , Male , Mice , Pregnancy , Sex Chromosomes/genetics
11.
Genome Biol ; 14(5): R43, 2013 May 25.
Article in English | MEDLINE | ID: mdl-23706135

ABSTRACT

BACKGROUND: Inter-individual epigenetic variation, due to genetic, environmental or random influences, is observed in many eukaryotic species. In mammals, however, the molecular nature of epiallelic variation has been poorly defined, partly due to the restricted focus on DNA methylation. Here we report the first genome-scale investigation of mammalian epialleles that integrates genomic, methylomic, transcriptomic and histone state information. RESULTS: First, in a small sample set, we demonstrate that non-genetically determined inter-individual differentially methylated regions (iiDMRs) can be temporally stable over at least 2 years. Then, we show that iiDMRs are associated with changes in chromatin state as measured by inter-individual differences in histone variant H2A.Z levels. However, the correlation of promoter iiDMRs with gene expression is negligible and not improved by integrating H2A.Z information. We find that most promoter epialleles, whether genetically or non-genetically determined, are associated with low levels of transcriptional activity, depleted for housekeeping genes, and either depleted for H3K4me3/enriched for H3K27me3 or lacking both these marks in human embryonic stem cells. The preferential enrichment of iiDMRs at regions of relative transcriptional inactivity validates in a larger independent cohort, and is reminiscent of observations previously made for promoters that undergo hypermethylation in various cancers, in vitro cell culture and ageing. CONCLUSIONS: Our work identifies potential key features of epiallelic variation in humans, including temporal stability of non-genetically determined epialleles, and concomitant perturbations of chromatin state. Furthermore, our work suggests a novel mechanistic link among inter-individual epialleles observed in the context of normal variation, cancer and ageing.


Subject(s)
Embryonic Stem Cells/metabolism , Epigenesis, Genetic , Genomics/methods , Promoter Regions, Genetic , Twins, Monozygotic/genetics , Alleles , DNA Methylation , Female , Gene Expression Regulation , Genome, Human , Histones/metabolism , Humans , Molecular Sequence Data
12.
Epigenetics ; 8(4): 445-54, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23538714

ABSTRACT

There is increasing evidence that interindividual epigenetic variation is an etiological factor in common human diseases. Such epigenetic variation could be genetic or non-genetic in origin, and epigenome-wide association studies (EWASs) are underway for a wide variety of diseases/phenotypes. However, performing an EWAS is associated with a range of issues not typically encountered in genome-wide association studies (GWASs), such as the tissue to be analyzed. In many EWASs, it is not possible to analyze the target tissue in large numbers of live humans, and consequently surrogate tissues are employed, most commonly blood. But there is as yet no evidence demonstrating that blood is more informative than buccal cells, the other easily accessible tissue. To assess the potential of buccal cells for use in EWASs, we performed a comprehensive analysis of a buccal cell methylome using whole-genome bisulfite sequencing. Strikingly, a buccal vs. blood comparison reveals>6X as many hypomethylated regions in buccal. These tissue-specific differentially methylated regions (tDMRs) are strongly enriched for DNaseI hotspots. Almost 75% of these tDMRs are not captured by commonly used DNA methylome profiling platforms such as Reduced Representational Bisulfite Sequencing and the Illumina Infinium HumanMethylation450 BeadChip, and they also display distinct genomic properties. Buccal hypo-tDMRs show a statistically significant enrichment near SNPs associated to disease identified through GWASs. Finally, we find that, compared with blood, buccal hypo-tDMRs show significantly greater overlap with hypomethylated regions in other tissues. We propose that for non-blood based diseases/phenotypes, buccal will be a more informative tissue for EWASs.


Subject(s)
DNA Methylation , DNA/metabolism , Epigenesis, Genetic , Mouth Mucosa/metabolism , Adult , Aged , DNA/blood , Genome, Human , Genome-Wide Association Study/methods , Humans , Middle Aged , Polymorphism, Single Nucleotide , Sequence Analysis, DNA , Young Adult
13.
Genome Res ; 22(11): 2138-45, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22919074

ABSTRACT

A major concern in common disease epigenomics is distinguishing causal from consequential epigenetic variation. One means of addressing this issue is to identify the temporal origins of epigenetic variants via longitudinal analyses. However, prospective birth-cohort studies are expensive and time consuming. Here, we report DNA methylomics of archived Guthrie cards for the retrospective longitudinal analyses of in-utero-derived DNA methylation variation. We first validate two methodologies for generating comprehensive DNA methylomes from Guthrie cards. Then, using an integrated epigenomic/genomic analysis of Guthrie cards and follow-up samplings, we identify interindividual DNA methylation variation that is present both at birth and 3 yr later. These findings suggest that disease-relevant epigenetic variation could be detected at birth, i.e., before overt clinical disease. Guthrie card methylomics offers a potentially powerful and cost-effective strategy for studying the dynamics of interindividual epigenomic variation in a range of common human diseases.


Subject(s)
Alleles , DNA Methylation , Epigenesis, Genetic , Female , Genetic Loci , Genetic Variation , Genome, Human , Hematologic Tests , High-Throughput Nucleotide Sequencing , Humans , Infant, Newborn , Longitudinal Studies , Male , Sequence Analysis, DNA
14.
Metabolism ; 60(8): 1142-9, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21310444

ABSTRACT

The regulation of mitochondrial DNA (mtDNA) copy number not only is critical for the maintenance of the normal mitochondrial function but has a strong clinical significance. A recent report revealed that the signal transducer and activator of transcription 3 (STAT3) is involved in the regulation of the mitochondrial function and is required for the optimal function of the electron transport chain. In this study, we explored whether gene variants in the STAT3 influence the leukocyte mtDNA copy number. Clinical data and blood samples were collected from 179 subjects (aged 52.8 ± 0.9 years). Mitochondrial DNA quantification using nuclear DNA (nDNA) as a reference was carried out by a real-time quantitative polymerase chain reaction method; results are presented as the mtDNA/nDNA ratio. We selected 3 tag single nucleotide polymorphisms showing a minor allele frequency greater than 10% (rs2293152 C/G, rs6503695 C/T, and rs9891119 A/C), representing 24 polymorphic sites of the STAT3 (r(2) > 0.8). We observed a significant association between mtDNA/nDNA ratio and both rs6503695 and rs9891119, adjusted by age and homeostasis model assessment index. The proportion of the total variance of the mtDNA/nDNA ratio accounted for by the rs6503695 and rs9891119 genotypes was 4.7% and 6.53%, respectively. Common variation in the STAT3 may influence mtDNA copy number.


Subject(s)
DNA, Mitochondrial/genetics , Genetic Variation , STAT3 Transcription Factor/genetics , Alleles , Female , Gene Frequency , Genotype , Humans , Male , Middle Aged , Mitochondria/genetics , Polymorphism, Single Nucleotide
15.
Hepatology ; 52(6): 1992-2000, 2010 Dec.
Article in English | MEDLINE | ID: mdl-20890895

ABSTRACT

UNLABELLED: Insulin resistance (IR) and mitochondrial dysfunction play a central role in the pathophysiology of nonalcoholic fatty liver disease (NAFLD). We hypothesized that genetic factors and epigenetic modifications occurring in the liver contribute to the IR phenotype. We specifically examined whether fatty liver and IR are modified by hepatic DNA methylation of the peroxisome proliferator-activated receptor γ coactivator 1α (PPARGC1A) and mitochondrial transcription factor A (TFAM) promoters, and also evaluated whether liver mitochondrial DNA (mtDNA) content is associated with NAFLD and IR. We studied liver biopsies obtained from NAFLD patients in a case-control design. After bisulfite treatment of DNA, we used methylation-specific polymerase chain reaction (PCR) to assess the putative methylation of three CpG in the PPARGC1A and TFAM promoters. Liver mtDNA quantification using nuclear DNA (nDNA) as a reference was evaluated by way of real-time PCR. Liver PPARGC1A methylated DNA/unmethylated DNA ratio correlated with plasma fasting insulin levels and homeostasis model assessment of insulin resistance (HOMA-IR); TFAM methylated DNA/unmethylated DNA ratio was inversely correlated with insulin levels. PPARGC1A promoter methylation was inversely correlated with the abundance of liver PPARGC1A messenger RNA. The liver mtDNA/nDNA ratio was significantly higher in control livers compared with NAFLD livers. mtDNA/nDNA ratio was inversely correlated with HOMA-IR, fasting glucose, and insulin and was inversely correlated with PPARGC1A promoter methylation. CONCLUSION: Our data suggest that the IR phenotype and the liver transcriptional activity of PPARGC1A show a tight interaction, probably through epigenetic modifications. Decreased liver mtDNA content concomitantly contributes to peripheral IR.


Subject(s)
Epigenesis, Genetic/physiology , Fatty Liver/genetics , Heat-Shock Proteins/genetics , Insulin Resistance/genetics , PPAR gamma/genetics , Promoter Regions, Genetic/genetics , Transcription Factors/genetics , Adult , Case-Control Studies , CpG Islands/genetics , DNA Methylation , DNA, Mitochondrial/metabolism , DNA-Binding Proteins/genetics , Female , Humans , Male , Middle Aged , Mitochondria, Liver/physiology , Mitochondrial Proteins/genetics , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha , RNA, Messenger/metabolism
16.
Mol Genet Metab ; 100(1): 83-7, 2010 May.
Article in English | MEDLINE | ID: mdl-20202876

ABSTRACT

PURPOSE: To explore whether DNA methylation of the mitochondrial transcription factor A (TFAM) promoter is associated with insulin resistance in a sample of adolescents with features of metabolic syndrome. METHODS: The data and blood samples were collected from 122 adolescents out of a cross-sectional study of 934 high-school students. The population was divided into two groups: noninsulin resistance (NIR) and insulin resistance (IR). After bisulfite treatment of genomic DNA from peripheral leukocytes, we used methylation-specific polymerase chain reaction (PCR) to assess DNA methylation of three putative methylation target sites (CpG) in the TFAM promoter. RESULTS: The ratio of the promoter methylated DNA/unmethylated DNA was 0.012+/-0.0009 (1.2% of alleles), and inversely correlated with the biochemical features of insulin resistance (plasma fasting insulin R: -0.26, p<0.004 and homeostasis model assessment (HOMA) index R: -0.27, p<0.002), and obesity (R: -0.27, p<0.002). Multiple regression analysis showed that the log-transformed HOMA index correlated with the status of promoter methylation of TFAM, independently of body mass index (BMI) Z score (beta: -0.33+/-0.094, p=0.00094). Finally, the TFAM promoter methylated DNA/unmethylated DNA ratio was found to be significantly associated with insulin resistance as dichotomous variable (NIR n=45, 0.014+/-0.002 and IR n=77, 0.011+/-0.001, respectively, p<0.016). CONCLUSION: Our findings suggest a potential role of promoter TFAM methylation in the pathogenesis of insulin resistance in adolescents.


Subject(s)
DNA-Binding Proteins/genetics , Insulin Resistance/genetics , Metabolic Syndrome/genetics , Mitochondrial Proteins/genetics , Promoter Regions, Genetic , Transcription Factors/genetics , Adolescent , Base Sequence , DNA Methylation , Female , Humans , Leukocytes/metabolism , Male , Molecular Sequence Data , Sp1 Transcription Factor/metabolism
17.
J Hypertens ; 28(6): 1194-202, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20179633

ABSTRACT

OBJECTIVE: To perform a two-stage study to explore the role of gene variants in the risk of insulin resistance and arterial hypertension. METHODS AND RESULTS: The selection of variants was performed by a first stage of in-silico analysis of the original genome-wide association data sets on genes involved in metabolic syndrome components, granted by the Diabetes Genetics Initiative and the Wellcome Trust Case-Control Consortium. We started by identifying single-nucleotide polymorphisms with a cutoff for association (P < 0.05) in both data sets after the application of a computational algorithm of gene prioritization. Among the more promising variants, six single-nucleotide polymorphisms in IGF1R (rs11247362, rs10902606, rs1317459, rs11854132, rs2684761, and rs2715416) were selected for further evaluation in our population. Altogether, 1094 men, aged 34.4 +/- 8.6 years, were included in a population-based study. Genotypes of rs2684761 showed significant association with insulin resistance (as a discrete trait, odds ratio per G allele 1.27, 95% confidence interval 1.03-1.56, P = 0.026; and homeostasis model assessment-insulin resistance as a continuous trait, P = 0.01). A significant association of rs2684761 with arterial hypertension was also observed (odds ratio per G allele 1.29, 95% confidence interval 1.02-1.64, P = 0.037) after adjusting for age and homeostasis model assessment-insulin resistance. CONCLUSION: Our study suggests for the first time a putative role of IGF1R variants in individual susceptibility to metabolic syndrome-related phenotypes, in particular on the risk of having insulin resistance and arterial hypertension.


Subject(s)
Genetic Variation , Hypertension/genetics , Insulin Resistance/genetics , Receptor, IGF Type 1/genetics , Female , Genotype , Humans , Linkage Disequilibrium , Male
18.
Atherosclerosis ; 210(2): 625-9, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20106477

ABSTRACT

OBJECTIVE: Shift work schedule has been associated with several health problems, including deleterious effects on the cardiovascular system. The present study aimed to evaluate the circulating levels of four biomarkers of atherosclerosis (soluble CD40 ligand [sCD40L], monocyte chemoattractant protein-1 [MCP-1], resistin, and plasminogen activator inhibitor-1 [PAI-1]) in a population-based sample of young adult men exposed to rotating shift work schedule in comparison with day workers. DESIGN AND PARTICIPANTS: A total of 439 men aged 34.4+/-8.6 years were included in a cross-sectional study comparing 255 day workers with 184 rotating shift workers. Circulating levels of the biomarkers were measured in duplicate by ELISA using monoclonal specific antibodies. RESULTS: Rotating shift workers had elevated (6440+/-4510 pg/mL) (mean+/-SD) circulating levels of resistin in comparison with day workers (5450+/-3780 pg/mL), and significance remains after adjusting for age and blood leukocyte count (p<0.045, ANCOVA). Shift work schedule explains 1% of the proportion of the total variation in the circulating resistin levels. Multiple regression analysis showed that resistin levels significantly correlate with rotating shift work (p<0.04) and blood leukocyte count (p<0.00003) independently of age, BMI, waist-hip ratio, HOMA, and cardiovascular risk %. Circulating levels of sCD40L, MCP-1, and PAI-1 did not significantly differ between day workers and shift workers. CONCLUSION: Shift work schedule was significantly associated with elevated plasma resistin levels. Resistin, which is probably produced by leukocytes, may play an important role in the pathogenesis of early metabolic syndrome components in young men chronically exposed to circadian misalignment.


Subject(s)
Cardiovascular Diseases/blood , Gene Expression Regulation , Resistin/blood , Work Schedule Tolerance , Adult , Antibodies, Monoclonal/metabolism , Atherosclerosis/blood , Atherosclerosis/diagnosis , CD40 Ligand/blood , Cardiovascular System , Chemokine CCL2/blood , Circadian Rhythm , Cross-Sectional Studies , Humans , Male , Plasminogen Activator Inhibitor 1/blood
19.
Mol Genet Metab ; 99(1): 80-9, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19748811

ABSTRACT

BACKGROUND: The nuclear receptor hepatocyte nuclear factor 4alpha (HNF4alpha) contributes to the regulation of a large fraction of liver and pancreatic islet transcriptomes. AIM: To evaluate the influence of HNF4alpha polymorphisms across the entire locus on the occurrence of type 2 diabetes (T2D) by means of a meta-analysis. METHODS: We evaluated haplotype block structure of HNF4alpha variants owing to linkage disequilibrium (LD). From 1455 reports, we evaluated 21 observational studies. RESULTS: Six haplotype blocks of LD were constructed with SNPs with r(2)>0.8; there were also 14 unlinked SNPs. Overall, we included 22,920 cases and 26.657 controls. Among 17 heterogeneous studies (21,881 cases and 24,915 controls), including 3 SNPs of P2 promoter region in block 1, we observed a significant association with T2D in fixed (OR 0.94, 95%CI: 0.905-0.975, p=0.001) and random (OR 0.988, 95%CI: 0.880-0.948, p=0.000012) model. Three homogeneous studies were evaluated in block 2 (2684 cases and 2059 controls), and a significant association with T2D was also observed: OR: 1.121, 95%CI 1.013-1.241, p=0.027. Three additional variants were associated with T2D: two intronic SNPs (rs4810424: OR: 1.080, 95%CI: 1.010-1.154, p<0.03 and rs3212183: OR: 0.843, 95%CI: 0.774-0.918, p<0.00009) and one missense variant (rs1800961: OR: 0.770, 95%CI: 0.595-0.995, p<0.05, 6562 cases and 6723 controls). CONCLUSIONS: In addition to HNF4alpha variants in the promoter region, other SNPs may be involved on the occurrence of T2D.


Subject(s)
Diabetes Mellitus, Type 2/genetics , Haplotypes , Hepatocyte Nuclear Factor 4/genetics , Polymorphism, Genetic , Genetic Predisposition to Disease , Genetic Variation , Genotype , Humans , Linkage Disequilibrium , Odds Ratio , Polymorphism, Single Nucleotide , Promoter Regions, Genetic/genetics , Risk Factors
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