1  5753 140 SOCIAL ISOLATION REDUCES MAMMARY DEVELOPMENT, TUMOR INCIDENCE, AND EXPRESSION OF EPIGENETIC REGULATORS IN WILD-TYPE AND P53-HETEROZYGOTIC MICE. CHRONIC STRESS IS ASSOCIATED WITH MORE RAPID TUMOR PROGRESSION, AND RECENT EVIDENCE SUGGESTS THAT STRESS MAY CONTRIBUTE TO SOCIAL AND ETHNIC DISPARITIES IN THE INCIDENCE AND MORTALITY OF BREAST CANCER. WE EVALUATED THE P53(+/-) FVB/N MOUSE AS A MODEL TO INVESTIGATE EFFECTS OF CHRONIC SOCIAL STRESS ON MAMMARY GLAND DEVELOPMENT, GENE EXPRESSION, AND TUMORIGENESIS. WE INDIVIDUALLY HOUSED (IH) WILD-TYPE AND P53(+/-) FEMALE FVB/N MICE, STARTING AT WEANING. AT 14 WEEKS OF AGE, BOTH WILD-TYPE AND P53(+/-) IH MICE SHOWED STRIKINGLY REDUCED MAMMARY DEVELOPMENT COMPARED WITH GROUP-HOUSED (GH) CONTROLS, WITH IH MICE HAVING SIGNIFICANTLY FEWER PRETERMINAL END BUDS. THIS MORPHOLOGIC DIFFERENCE WAS NOT REFLECTED IN LEVELS OF MAMMARY TRANSCRIPTS FOR ESTROGEN RECEPTOR-ALPHA OR PROGESTIN RECEPTOR. HOWEVER, IH INCREASED LEVELS OF MRNA FOR THE KISSPEPTIN RECEPTOR IN THE MEDIAL PREOPTIC AREA OF THE HYPOTHALAMUS, ASSOCIATED WITH REDUCED DURATION OF ESTROUS CYCLES. FURTHERMORE, IH ALTERED MAMMARY TRANSCRIPTS OF GENES ASSOCIATED WITH DNA METHYLATION; TRANSCRIPTS FOR METHYL-BINDING PROTEIN 2 AND DNA METHYLTRANSFERASE 3B (DNMT3B), BUT NOT DNMT1 AND DNMT3A, WERE REDUCED IN IH COMPARED WITH GH FEMALES. INTERESTINGLY, THE GLANDS OF P53(+/-) FEMALES SHOWED REDUCED EXPRESSION OF ALL THESE MEDIATORS COMPARED WITH WILD-TYPE FEMALES. HOWEVER, CONTRARY TO OUR INITIAL HYPOTHESIS, IH DID NOT INCREASE MAMMARY TUMORIGENESIS. RATHER, P53(+/-) GH FEMALES DEVELOPED SIGNIFICANTLY MORE MAMMARY TUMORS THAN IH MICE. TOGETHER, THESE DATA SUGGEST THAT SOCIAL ISOLATION INITIATED AT PUBERTY MIGHT CONFOUND STUDIES OF TUMORIGENESIS BY ALTERING MAMMARY DEVELOPMENT IN MOUSE MODELS.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
2  4811  47 OBESITY-ASSOCIATED ALTERATIONS IN INFLAMMATION, EPIGENETICS, AND MAMMARY TUMOR GROWTH PERSIST IN FORMERLY OBESE MICE. USING A MURINE MODEL OF BASAL-LIKE BREAST CANCER, WE TESTED THE HYPOTHESIS THAT CHRONIC OBESITY, AN ESTABLISHED BREAST CANCER RISK AND PROGRESSION FACTOR IN WOMEN, INDUCES MAMMARY GLAND EPIGENETIC REPROGRAMMING AND INCREASES MAMMARY TUMOR GROWTH. MOREOVER, WE ASSESSED WHETHER THE OBESITY-INDUCED EPIGENETIC AND PROTUMOR EFFECTS ARE REVERSED BY WEIGHT NORMALIZATION. OVARIECTOMIZED FEMALE C57BL/6 MICE WERE FED A CONTROL DIET OR DIET-INDUCED OBESITY (DIO) REGIMEN FOR 17 WEEKS, RESULTING IN A NORMAL WEIGHT OR OBESE PHENOTYPE, RESPECTIVELY. MICE ON THE DIO REGIMEN WERE THEN RANDOMIZED TO CONTINUE THE DIO DIET OR WERE SWITCHED TO THE CONTROL DIET, RESULTING IN FORMERLY OBESE (FOB) MICE WITH WEIGHTS COMPARABLE WITH CONTROL MICE. AT WEEK 24, ALL MICE WERE ORTHOTOPICALLY INJECTED WITH MMTV-WNT-1 MOUSE MAMMARY TUMOR CELLS. MEAN TUMOR VOLUME, SERUM IL6 LEVELS, EXPRESSION OF PROINFLAMMATORY GENES IN THE MAMMARY FAT PAD, AND MAMMARY DNA METHYLATION PROFILES WERE SIMILAR IN DIO AND FOB MICE AND HIGHER THAN IN CONTROLS. MANY OF THE GENES FOUND TO HAVE OBESITY-ASSOCIATED HYPERMETHYLATION IN MICE WERE ALSO FOUND TO BE HYPERMETHYLATED IN THE NORMAL BREAST TISSUE OF OBESE VERSUS NONOBESE HUMAN SUBJECTS, AND NEARLY ALL OF THESE CONCORDANT GENES REMAINED HYPERMETHYLATED AFTER SIGNIFICANT WEIGHT LOSS IN THE FOB MICE. OUR FINDINGS SUGGEST THAT WEIGHT NORMALIZATION MAY NOT BE SUFFICIENT TO REVERSE THE EFFECTS OF CHRONIC OBESITY ON EPIGENETIC REPROGRAMMING AND INFLAMMATORY SIGNALS IN THE MICROENVIRONMENT THAT ARE ASSOCIATED WITH BREAST CANCER PROGRESSION. CANCER PREV RES; 9(5); 339-48. (C)2016 AACR.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
3  5187  36 PRENATAL AIRBORNE POLYCYCLIC AROMATIC HYDROCARBON EXPOSURE, ALTERED REGULATION OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR GAMMA (PPAR)GAMMA, AND LINKS WITH MAMMARY CANCER. ENVIRONMENTAL EXPOSURE TO POLYCYCLIC AROMATIC HYDROCARBONS (PAH) HAS BEEN SHOWN TO BE ASSOCIATED WITH CHRONIC DISEASE OUTCOMES THROUGH MULTIPLE MECHANISMS INCLUDING ALTERED REGULATION OF THE TRANSCRIPTION FACTOR PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR GAMMA (PPAR) GAMMA. BECAUSE PAH EXPOSURE AND PPARGAMMA EACH HAVE BEEN ASSOCIATED WITH MAMMARY CANCER, WE ASKED WHETHER PAH WOULD INDUCE ALTERED REGULATION OF PPARGAMMA IN MAMMARY TISSUE, AND WHETHER THIS ASSOCIATION MAY UNDERLIE THE ASSOCIATION BETWEEN PAH AND MAMMARY CANCER. PREGNANT MICE WERE EXPOSED TO AEROSOLIZED PAH AT PROPORTIONS THAT MIMIC EQUIVALENT HUMAN EXPOSURES IN NEW YORK CITY AIR. WE HYPOTHESIZED THAT PRENATAL PAH EXPOSURE WOULD ALTER PPARGAMMA DNA METHYLATION AND GENE EXPRESSION AND INDUCE THE EPITHELIAL TO MESENCHYMAL TRANSITION (EMT) IN MAMMARY TISSUE OF OFFSPRING (F1) AND GRANDOFFSPRING (F2) MICE. WE ALSO HYPOTHESIZED THAT ALTERED REGULATION OF PPARGAMMA IN MAMMARY TISSUE WOULD ASSOCIATE WITH BIOMARKERS OF EMT, AND EXAMINED ASSOCIATIONS WITH WHOLE BODY WEIGHT. WE FOUND THAT PRENATAL PAH EXPOSURE LOWERED PPARGAMMA MAMMARY TISSUE METHYLATION AMONG GRANDOFFSPRING MICE AT POSTNATAL DAY (PND) 28. HOWEVER, PAH EXPOSURE DID NOT ASSOCIATE WITH ALTERED PPARGAMMA GENE EXPRESSION OR CONSISTENTLY WITH BIOMARKERS OF EMT. FINALLY, LOWER PPARGAMMA METHYLATION, BUT NOT GENE EXPRESSION, WAS ASSOCIATED WITH HIGHER BODY WEIGHT AMONG OFFSPRING AND GRANDOFFSPRING MICE AT PND28 AND PND60. FINDINGS SUGGEST ADDITIONAL EVIDENCE OF MULTI-GENERATIONAL ADVERSE EPIGENETIC EFFECTS OF PRENATAL PAH EXPOSURE AMONG GRANDOFFSPRING MICE.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
4  1899  45 ENERGY BALANCE MODULATION IMPACTS EPIGENETIC REPROGRAMMING, ERALPHA AND ERBETA EXPRESSION, AND MAMMARY TUMOR DEVELOPMENT IN MMTV-NEU TRANSGENIC MICE. THE ASSOCIATION BETWEEN OBESITY AND BREAST CANCER RISK AND PROGNOSIS IS WELL ESTABLISHED IN ESTROGEN RECEPTOR (ER)-POSITIVE DISEASE BUT LESS CLEAR IN HER2-POSITIVE DISEASE. HERE, WE REPORT PRECLINICAL EVIDENCE SUGGESTING WEIGHT MAINTENANCE THROUGH CALORIE RESTRICTION (CR) MAY LIMIT RISK OF HER2-POSITIVE BREAST CANCER. IN FEMALE MMTV-HER2/NEU TRANSGENIC MICE, WE FOUND THAT ERALPHA AND ERBETA EXPRESSION, MAMMARY TUMORIGENESIS, AND SURVIVAL ARE ENERGY BALANCE DEPENDENT IN ASSOCIATION WITH EPIGENETIC REPROGRAMMING. MICE WERE RANDOMIZED TO RECEIVE A CR, OVERWEIGHT-INDUCING, OR DIET-INDUCED OBESITY REGIMEN (N = 27/GROUP). SUBSETS OF MICE (N = 4/GROUP/TIME POINT) WERE EUTHANIZED AFTER 1, 3, AND 5 MONTHS TO CHARACTERIZE DIET-DEPENDENT METABOLIC, TRANSCRIPTIONAL, AND EPIGENETIC PERTURBATIONS. REMAINING MICE WERE FOLLOWED UP TO 22 MONTHS. RELATIVE TO THE OVERWEIGHT AND DIET-INDUCED OBESITY REGIMENS, CR DECREASED BODY WEIGHT, ADIPOSITY, AND SERUM METABOLIC HORMONES AS EXPECTED AND ALSO ELICITED AN INCREASE IN MAMMARY ERALPHA AND ERBETA EXPRESSION. INCREASED DNA METHYLATION ACCOMPANIED THIS PATTERN, PARTICULARLY AT CPG DINUCLEOTIDES LOCATED WITHIN BINDING OR FLANKING REGIONS FOR THE TRANSCRIPTIONAL REGULATOR CCCTC-BINDING FACTOR OF ESR1 AND ESR2, CONSISTENT WITH SUSTAINED TRANSCRIPTIONAL ACTIVATION OF ERALPHA AND ERBETA. MAMMARY EXPRESSION OF THE DNA METHYLATION ENZYME DNMT1 WAS STABLE IN CR MICE BUT INCREASED OVER TIME IN OVERWEIGHT AND DIET-INDUCED OBESITY MICE, SUGGESTING CR OBVIATES EPIGENETIC ALTERATIONS CONCURRENT WITH CHRONIC EXCESS ENERGY INTAKE. IN THE SURVIVAL STUDY, CR ELICITED A SIGNIFICANT SUPPRESSION IN SPONTANEOUS MAMMARY TUMORIGENESIS. OVERALL, OUR FINDINGS SUGGEST A MECHANISTIC RATIONALE TO PREVENT OR REVERSE EXCESS BODY WEIGHT AS A STRATEGY TO REDUCE HER2-POSITIVE BREAST CANCER RISK. CANCER RES; 77(9); 2500-11. (C)2017 AACR.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
5  2749  37 EXPRESSION AND POLYMORPHISM OF MICRO-RNA ACCORDING TO BODY MASS INDEX AND BREAST CANCER PRESENTATION IN TUNISIAN PATIENTS. MICRO-RNAS (MIRS) CONSTITUTE A CLASS OF SMALL NONCODING RNAS IMPLICATED IN THE REGULATION OF GENE EXPRESSION BY BINDING TO TARGET MRNAS. A MIR CAN TARGET SEVERAL MRNAS, BEING INVOLVED IN DIFFERENT BIOLOGIC PROCESSES AND PATHOLOGIES. THIS PLEIOTROPIC FUNCTION MIGHT EXPLAIN THE LINK BETWEEN DISEASES CO-OCCURRENCE. EPIGENETIC ORIGIN OF THE LINK BETWEEN OBESITY AND BREAST CANCER (BC) IS INVESTIGATED IN A COHORT OF TUNISIAN PATIENTS, FOCUSING ON POLYMORPHISM AT GERMLINE LEVEL (MIR-146A) AND ON EXPRESSION IN MAMMARY TUMORS (MIR-21, MIR-146A, AND MIR-34A), ACCORDING TO BODY MASS INDEX (BMI) AND CLINICO-PATHOLOGIC FEATURES. THE MEASURE OF MIR EXPRESSION IN 60 MAMMARY TUMORS WAS REALIZED USING QUANTITATIVE RT-PCR. STUDY OF RS 2910164 IN MIR-146A WAS PERFORMED BY PCR AND DIRECT SEQUENCING USING BLOOD DNA OF 83 AFFECTED WOMEN AND 50 UNRELATED SUBJECTS FROM GREAT TUNIS. MIR-21, MIR-146A, AND MIR-34A HAVE BEEN QUANTIFIED IN BREAST TUMOR ACCORDING TO BMI. MIR-21 IS SIGNIFICANTLY MORE EXPRESSED IN TUMORS OF OBESE WOMEN COMPARATIVELY TO NONOBESE PATIENTS. ON THE CONTRARY, MIR-34A IS DECREASED IN TUMORS OF OBESE WOMEN. MOREOVER, IN OBESE BC PATIENTS, A SIGNIFICANT INCREASE IN BOTH MIR-21 AND MIR-146A EXPRESSION IS REVEALED IN CASES WITH LYMPH NODE METASTASIS. THE POLYMORPHISM AT RS 2910164 (MIR-146A) LOCUS WAS NOT SHOWN AS A RISK FACTOR FOR BC. HOWEVER THE MUTANT CC GENOTYPE WAS REVEALED TO BE ASSOCIATED WITH A RISK FOR BAD OUTCOME OF THE DISEASE. CHRONIC INFLAMMATION IN OBESE WOMEN WOULD BE LINKED TO AGGRESSIVE BREAST TUMORS VIA INDUCTION OF ONCOMIRS OVEREXPRESSION AND DECREASE OF TUMOR SUPPRESSOR MIRS.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
6  3410  39 HOXA5 UNDERGOES DYNAMIC DNA METHYLATION AND TRANSCRIPTIONAL REPRESSION IN THE ADIPOSE TISSUE OF MICE EXPOSED TO HIGH-FAT DIET. BACKGROUND/OBJECTIVES: THE GENOMIC BASES OF THE ADIPOSE TISSUE ABNORMALITIES INDUCED BY CHRONIC POSITIVE CALORIE EXCESS HAVE BEEN ONLY PARTIALLY ELUCIDATED. WE ADOPTED A GENOME-WIDE APPROACH TO DIRECTLY TEST WHETHER LONG-TERM HIGH-FAT DIET (HFD) EXPOSURE AFFECTS THE DNA METHYLATION PROFILE OF THE MOUSE ADIPOSE TISSUE AND TO IDENTIFY THE FUNCTIONAL CONSEQUENCES OF THESE CHANGES. SUBJECTS/METHODS: WE HAVE USED EPIDIDYMAL FAT OF MICE FED EITHER HIGH-FAT (HFD) OR REGULAR CHOW (STD) DIET FOR 5 MONTHS AND PERFORMED GENOME-WIDE DNA METHYLATION ANALYSES BY METHYLATED DNA IMMUNOPRECIPITATION SEQUENCING (MEDIP-SEQ). MOUSE HOMEOBOX (HOX) GENE DNA METHYLATION PCR, RT-QPCR AND BISULPHITE SEQUENCING ANALYSES WERE THEN PERFORMED. RESULTS: MICE FED THE HFD PROGRESSIVELY EXPANDED THEIR ADIPOSE MASS ACCOMPANIED BY A SIGNIFICANT DECREASE IN GLUCOSE TOLERANCE (P<0.001) AND INSULIN SENSITIVITY (P<0.05). MEDIP-SEQ DATA ANALYSIS REVEALED A UNIFORM DISTRIBUTION OF DIFFERENTIALLY METHYLATED REGIONS (DMR) THROUGH THE ENTIRE ADIPOCYTE GENOME, WITH A HIGHER NUMBER OF HYPERMETHYLATED REGIONS IN HFD MICE (P<0.005). THIS DIFFERENT METHYLATION PROFILE WAS ACCOMPANIED BY INCREASED EXPRESSION OF THE DNMT3A DNA METHYLTRANSFERASE (DNMT; P<0.05) AND THE METHYL-CPG-BINDING DOMAIN PROTEIN MBD3 (P<0.05) GENES IN HFD MICE. GENE ONTOLOGY ANALYSIS REVEALED THAT, IN THE HFD-TREATED MICE, THE HOX FAMILY OF DEVELOPMENT GENES WAS HIGHLY ENRICHED IN DIFFERENTIALLY METHYLATED GENES (P=0.008). TO VALIDATE THIS FINDING, HOXA5, WHICH IS IMPLICATED IN FAT TISSUE DIFFERENTIATION AND REMODELING, HAS BEEN SELECTED AND ANALYZED BY BISULPHITE SEQUENCING, CONFIRMING HYPERMETHYLATION IN THE ADIPOSE TISSUE FROM THE HFD MICE. HOXA5 HYPERMETHYLATION WAS ASSOCIATED WITH DOWNREGULATION OF HOXA5 MRNA AND PROTEIN EXPRESSION. FEEDING ANIMALS PREVIOUSLY EXPOSED TO THE HFD WITH A STANDARD CHOW DIET FOR TWO FURTHER MONTHS IMPROVED THE METABOLIC PHENOTYPE OF THE ANIMALS, ACCOMPANIED BY RETURN OF HOXA5 METHYLATION AND EXPRESSION LEVELS (P<0.05) TO VALUES SIMILAR TO THOSE OF THE CONTROL MICE MAINTAINED UNDER STANDARD CHOW. CONCLUSIONS: HFD INDUCES ADIPOSE TISSUE ABNORMALITIES ACCOMPANIED BY EPIGENETIC CHANGES AT THE HOXA5 ADIPOSE TISSUE REMODELING GENE.	2016	
                                                                                                                                                                                                                                                                                                               
7  2405  35 EPIGENETIC RESPONSE IN MICE MASTITIS: ROLE OF HISTONE H3 ACETYLATION AND MICRORNA(S) IN THE REGULATION OF HOST INFLAMMATORY GENE EXPRESSION DURING STAPHYLOCOCCUS AUREUS INFECTION. BACKGROUND: THERE IS RENEWED INTEREST TOWARDS UNDERSTANDING THE HOST-PATHOGEN INTERACTION IN THE LIGHT OF EPIGENETIC MODIFICATIONS. ALTHOUGH EPITHELIAL TISSUE IS THE MAJOR SITE FOR HOST-PATHOGEN INTERACTIONS, THERE IS HANDFUL OF STUDIES TO SHOW HOW EPITHELIAL CELLS RESPOND TO PATHOGENS. BACTERIAL INFECTION IN THE MAMMARY GLAND PARENCHYMA INDUCES LOCAL AND SUBSEQUENTLY SYSTEMIC INFLAMMATION THAT RESULTS IN A COMPLEX DISEASE CALLED MASTITIS. GLOBALLY STAPHYLOCOCCUS AUREUS IS THE SINGLE LARGEST MASTITIS PATHOGEN AND THE INFECTION CAN ULTIMATELY RESULT IN EITHER SUBCLINICAL OR CHRONIC AND SOMETIMES LIFELONG INFECTION. RESULTS: IN THE PRESENT REPORT WE HAVE ADDRESSED THE DIFFERENTIAL INFLAMMATORY RESPONSE IN MICE MAMMARY TISSUE DURING INTRAMAMMARY INFECTION AND THE ALTERED EPIGENETIC CONTEXT INDUCED BY TWO CLOSELY RELATED STRAINS OF S. AUREUS, ISOLATED FROM FIELD SAMPLES. IMMUNOHISTOCHEMICAL AND IMMUNOBLOTTING ANALYSIS SHOWED STRAIN SPECIFIC HYPERACETYLATION AT HISTONE H3K9 AND H3K14 RESIDUES. GLOBAL GENE EXPRESSION ANALYSIS IN S. AUREUS INFECTED MICE MAMMARY TISSUE REVEALED A SELECTIVE SET OF UPREGULATED GENES THAT SIGNIFICANTLY CORRELATED WITH THE PROMOTER SPECIFIC, HISTONE H3K14 ACETYLATION. FURTHERMORE, WE HAVE IDENTIFIED SEVERAL DIFFERENTIALLY EXPRESSED KNOWN MIRNAS AND 3 NOVEL MIRNAS IN S. AUREUS INFECTED MICE MAMMARY TISSUE BY SMALL RNA SEQUENCING. BY EMPLOYING THESE GENE EXPRESSION DATA, AN ATTEMPT HAS BEEN MADE TO DELINEATE THE GENE REGULATORY NETWORKS IN THE STRAIN SPECIFIC INFLAMMATORY RESPONSE. APPARENTLY, ONE OF THE ISOLATES OF S. AUREUS ACTIVATED THE NF-KAPPAB SIGNALING LEADING TO DRASTIC INFLAMMATORY RESPONSE AND INDUCTION OF IMMUNE SURVEILLANCE, WHICH COULD POSSIBLY LEAD TO RAPID CLEARANCE OF THE PATHOGEN. THE OTHER STRAIN REPRESSED MOST OF THE INFLAMMATORY RESPONSE, WHICH MIGHT HELP IN ITS SUSTENANCE IN THE HOST TISSUE. CONCLUSION: TAKEN TOGETHER, OUR STUDIES SHED SUBSTANTIAL LIGHTS TO UNDERSTAND THE MECHANISMS OF STRAIN SPECIFIC DIFFERENTIAL INFLAMMATORY RESPONSE TO S. AUREUS INFECTION DURING MASTITIS. IN A BROADER PERSPECTIVE THIS STUDY ALSO PAVES THE WAY TO UNDERSTAND HOW CERTAIN BACTERIA CAN EVADE THE IMMUNE SURVEILLANCE AND CAUSE SUSTAINED INFECTION WHILE OTHERS ARE RAPIDLY CLEARED FROM THE HOST BODY.	2014	
                                                                                                                                                                                                                    
8  1620  36 DNA METHYLTRANSFERASE-MEDIATED TRANSCRIPTIONAL SILENCING IN MALIGNANT GLIOMA: A COMBINED WHOLE-GENOME MICROARRAY AND PROMOTER ARRAY ANALYSIS. EPIGENETIC INACTIVATION OF TUMOR SUPPRESSOR GENES IS A COMMON FEATURE IN HUMAN CANCER. PROMOTER HYPERMETHYLATION AND HISTONE DEACETYLATION ARE REVERSIBLE EPIGENETIC MECHANISMS ASSOCIATED WITH TRANSCRIPTIONAL REGULATION. DNA METHYLTRANSFERASES (DNMT1 AND DNMT3B) REGULATE AND MAINTAIN PROMOTER METHYLATION AND ARE OVEREXPRESSED IN HUMAN CANCER. WE PERFORMED WHOLE-GENOME MICROARRAY ANALYSIS TO IDENTIFY GENES WITH ALTERED EXPRESSION AFTER RNAI-INDUCED SUPPRESSION OF DNMT IN A GLIOBLASTOMA MULTIFORME (GBM) CELL LINE. WE THEN IDENTIFIED GENES WITH BOTH DECREASED EXPRESSION AND EVIDENCE OF PROMOTER CPG ISLAND HYPERMETHYLATION IN GBM TISSUE SAMPLES USING A COMBINED WHOLE-GENOME MICROARRAY TRANSCRIPTOME ANALYSIS IN CONJUNCTION WITH A PROMOTER ARRAY ANALYSIS AFTER DNA IMMUNOPRECIPITATION WITH ANTI-5-METHYLCYTIDINE. DNMT1 AND 3B KNOCKDOWN RESULTED IN THE RESTORED EXPRESSION OF 308 GENES THAT ALSO CONTAINED PROMOTER REGION HYPERMETHYLATION. OF THESE, 43 WERE ALSO FOUND TO BE DOWNREGULATED IN GBM TISSUE SAMPLES. THREE DOWNREGULATED GENES WITH HYPERMETHYLATED PROMOTERS AND RESTORED EXPRESSION IN RESPONSE TO ACUTE DNMT SUPPRESSION WERE ASSAYED FOR METHYLATION CHANGES USING BISULFITE SEQUENCE ANALYSIS OF THE PROMOTER REGION AFTER CHRONIC DNMT SUPPRESSION. RESTORATION OF GENE EXPRESSION WAS NOT ASSOCIATED WITH CHANGES IN PROMOTER REGION METHYLATION, BUT RATHER WITH CHANGES IN HISTONE METHYLATION AND CHROMATIN CONFORMATION. TWO OF THE IDENTIFIED GENES EXHIBITED GROWTH SUPPRESSIVE ACTIVITY IN IN VITRO ASSAYS. COMBINING TARGETED GENETIC MANIPULATIONS WITH COMPREHENSIVE GENOMIC AND EXPRESSION ANALYSES PROVIDES A POTENTIALLY POWERFUL NEW APPROACH FOR IDENTIFYING EPIGENETICALLY REGULATED GENES IN GBM.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
9   286  39 AGING AND ALCOHOL INTERACT TO ALTER HEPATIC DNA HYDROXYMETHYLATION. BACKGROUND: AGING AND CHRONIC ALCOHOL CONSUMPTION ARE BOTH MODIFIERS OF DNA METHYLATION, BUT IT IS NOT YET KNOWN WHETHER CHRONIC ALCOHOL CONSUMPTION ALSO ALTERS DNA HYDROXYMETHYLATION, A NEWLY DISCOVERED EPIGENETIC MARK PRODUCED BY OXIDATION OF METHYLCYTOSINE. FURTHERMORE, IT HAS NOT BEEN TESTED WHETHER AGING AND ALCOHOL INTERACT TO MODIFY THIS EPIGENETIC PHENOMENON, THEREBY HAVING AN INDEPENDENT EFFECT ON GENE EXPRESSION. METHODS: OLD (18 MONTHS) AND YOUNG (4 MONTHS) MALE C57BL/6 MICE WERE PAIR-FED EITHER A LIEBER-DECARLI LIQUID DIET WITH ALCOHOL (18% OF ENERGY) OR AN ISOCALORIC LIEBER-DECARLI CONTROL DIET FOR 5 WEEKS. GLOBAL DNA HYDROXYMETHYLATION AND DNA METHYLATION WERE ANALYZED FROM HEPATIC DNA USING A NEW LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY METHOD. HEPATIC MRNA EXPRESSION OF THE TET ENZYMES WERE MEASURED VIA QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION. RESULTS: IN YOUNG MICE, MILD CHRONIC ALCOHOL EXPOSURE SIGNIFICANTLY REDUCED GLOBAL DNA HYDROXYMETHYLATION COMPARED WITH CONTROL MICE (0.22 +/- 0.01 VS. 0.29 +/- 0.06%, P = 0.004). ALCOHOL DID NOT SIGNIFICANTLY ALTER HYDROXYMETHYLCYTOSINE LEVELS IN OLD MICE. OLD MICE FED THE CONTROL DIET SHOWED DECREASED GLOBAL DNA HYDROXYMETHYLATION COMPARED WITH YOUNG MICE FED THE CONTROL DIET (0.24 +/- 0.02 VS. 0.29 +/- 0.06%, P = 0.04). THIS MODEL SUGGESTS AN INTERACTION BETWEEN AGING AND ALCOHOL IN DETERMINING DNA HYDROXYMETHYLATION (PINTERACTION = 0.009). EXPRESSION OF TET2 AND TET3 WAS DECREASED IN THE OLD MICE RELATIVE TO THE YOUNG (P < 0.005). CONCLUSIONS: THE OBSERVATION THAT ALCOHOL ALTERS DNA HYDROXYMETHYLATION INDICATES A NEW EPIGENETIC EFFECT OF ALCOHOL. THIS IS THE FIRST STUDY DEMONSTRATING THE INTERACTIVE EFFECTS OF CHRONIC ALCOHOL CONSUMPTION AND AGING ON DNA HYDROXYMETHYLATION.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
10  5744  28 SMOKING-INDUCED EXPRESSION OF THE GPR15 GENE INDICATES ITS POTENTIAL ROLE IN CHRONIC INFLAMMATORY PATHOLOGIES. DESPITE THE DESCRIBED CLEAR EPIGENETIC EFFECTS OF SMOKING, THE EFFECT OF SMOKING ON GENOME-WIDE GENE EXPRESSION IN THE BLOOD IS OBSCURE. WE THEREFORE STUDIED THE SMOKING-INDUCED CHANGES IN THE GENE-EXPRESSION PROFILE OF THE PERIPHERAL BLOOD. RNA WAS EXTRACTED FROM THE WHOLE BLOOD OF 48 INDIVIDUALS WITH A DETAILED SMOKING HISTORY (24 NEVER-SMOKERS, 16 SMOKERS, AND 8 EX-SMOKERS). GENE-EXPRESSION PROFILES WERE EVALUATED WITH RNA SEQUENCING, AND RESULTS WERE ANALYZED SEPARATELY IN 24 MEN AND 24 WOMEN. IN THE MALE SMOKERS, 13 GENES WERE STATISTICALLY SIGNIFICANTLY (FALSE-DISCOVERY RATE <0.1) DIFFERENTIALLY EXPRESSED; IN FEMALE SMOKERS, 5 GENES. ALTHOUGH MOST OF THE DIFFERENTIALLY EXPRESSED GENES WERE DIFFERENT BETWEEN THE MALE AND FEMALE SMOKERS, THE G-PROTEIN-COUPLED RECEPTOR 15 GENE (GPR15) WAS DIFFERENTIALLY EXPRESSED IN BOTH MALE AND FEMALE SMOKERS COMPARED WITH NEVER-SMOKERS. ANALYSIS OF GPR15 METHYLATION IDENTIFIED SIGNIFICANTLY GREATER HYPOMETHYLATION IN SMOKERS COMPARED WITH THAT IN NEVER-SMOKERS. GPR15 IS THE CHEMOATTRACTANT RECEPTOR THAT REGULATES T-CELL MIGRATION AND IMMUNITY. UP-REGULATION OF GPR15 COULD EXPLAIN TO SOME EXTENT THE HEALTH HAZARDS OF SMOKING WITH REGARD TO CHRONIC INFLAMMATORY DISEASES.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
11  1161  47 CONTINUOUS DEVELOPMENTAL AND EARLY LIFE TRICHLOROETHYLENE EXPOSURE PROMOTED DNA METHYLATION ALTERATIONS IN POLYCOMB PROTEIN BINDING SITES IN EFFECTOR/MEMORY CD4(+) T CELLS. TRICHLOROETHYLENE (TCE) IS AN INDUSTRIAL SOLVENT AND DRINKING WATER POLLUTANT ASSOCIATED WITH CD4(+) T CELL-MEDIATED AUTOIMMUNITY. IN OUR MOUSE MODEL, DISCONTINUATION OF TCE EXPOSURE DURING ADULTHOOD AFTER DEVELOPMENTAL EXPOSURE DID NOT PREVENT IMMUNOTOXICITY. TO DETERMINE WHETHER PERSISTENT EFFECTS WERE LINKED TO EPIGENETIC CHANGES WE CONDUCTED WHOLE GENOME REDUCED REPRESENTATION BISULFITE SEQUENCING (RRBS) TO EVALUATE METHYLATION OF CPG SITES IN AUTOSOMAL CHROMOSOMES IN ACTIVATED EFFECTOR/MEMORY CD4(+) T CELLS. FEMALE MRL+/+ MICE WERE EXPOSED TO VEHICLE CONTROL OR TCE IN THE DRINKING WATER FROM GESTATION UNTIL ~37 WEEKS OF AGE [POSTNATAL DAY (PND) 259]. IN A SUBSET OF MICE, TCE EXPOSURE WAS DISCONTINUED AT ~22 WEEKS OF AGE (PND 154). AT PND 259, RRBS ASSESSMENT REVEALED MORE GLOBAL METHYLATION CHANGES IN THE CONTINUOUS EXPOSURE GROUP VS. THE DISCONTINUOUS EXPOSURE GROUP. A MAJORITY OF THE DIFFERENTIALLY METHYLATED CPG REGIONS (DMRS) ACROSS PROMOTERS, ISLANDS, AND REGULATORY ELEMENTS WERE HYPERMETHYLATED (~90%). HOWEVER, CONTINUOUS DEVELOPMENTAL TCE EXPOSURE ALTERED THE METHYLATION OF 274 CPG SITES IN PROMOTERS AND CPG ISLANDS. IN CONTRAST, ONLY 4 CPG ISLAND REGIONS WERE DIFFERENTIALLY METHYLATED (HYPERMETHYLATED) IN THE DISCONTINUOUS GROUP. INTERESTINGLY, 2 OF THESE 4 SITES WERE ALSO HYPERMETHYLATED IN THE CONTINUOUS EXPOSURE GROUP, AND BOTH OF THESE ISLAND REGIONS ARE ASSOCIATED WITH LYSINE 27 ON HISTONE H3 (H3K27) INVOLVED IN POLYCOMB COMPLEX-DEPENDENT TRANSCRIPTIONAL REPRESSION VIA H3K27 TRI-METHYLATION. CPG SITES WERE OVERLAPPED WITH THE OPEN REGULATORY ANNOTATION DATABASE. UNLIKE THE DISCONTINUOUS GROUP, CONTINUOUS TCE TREATMENT RESULTED IN 129 DMRS INCLUDING 12 UNIQUE TRANSCRIPTION FACTORS AND REGULATORY ELEMENTS; 80% OF WHICH WERE ENRICHED FOR ONE OR MORE POLYCOMB GROUP (PCG) PROTEIN BINDING REGIONS (I.E., SUZ12, EZH2, JARID2, AND MTF2). PATHWAY ANALYSIS OF THE DMRS INDICATED THAT TCE PRIMARILY ALTERED THE METHYLATION OF GENES ASSOCIATED WITH REGULATION OF CELLULAR METABOLISM AND CELL SIGNALING. THE RESULTS DEMONSTRATED THAT CONTINUOUS DEVELOPMENTAL EXPOSURE TO TCE DIFFERENTIALLY METHYLATED BINDING SITES OF PCG PROTEINS IN EFFECTOR/MEMORY CD4(+) CELLS. THERE WERE MINIMAL YET POTENTIALLY BIOLOGICALLY SIGNIFICANT EFFECTS THAT OCCURRED WHEN EXPOSURE WAS DISCONTINUED. THESE RESULTS POINT TOWARD A NOVEL MECHANISM BY WHICH CHRONIC DEVELOPMENTAL TCE EXPOSURE MAY ALTER TERMINALLY DIFFERENTIATED CD4(+) T CELL FUNCTION IN ADULTHOOD.	2019	

12  1784  43 EFFECT OF ALCOHOL CONSUMPTION ON CPG METHYLATION IN THE DIFFERENTIALLY METHYLATED REGIONS OF H19 AND IG-DMR IN MALE GAMETES: IMPLICATIONS FOR FETAL ALCOHOL SPECTRUM DISORDERS. BACKGROUND: EXPOSURE TO ALCOHOL IN UTERO IS THE MAIN ATTRIBUTABLE CAUSE OF FETAL ALCOHOL SPECTRUM DISORDERS (FASD) WHICH IN ITS MOST SEVERE FORM IS CHARACTERIZED BY IRREVERSIBLE BEHAVIORAL AND COGNITIVE DISABILITY. PATERNAL PRECONCEPTION DRINKING IS NOT CONSIDERED TO BE A SIGNIFICANT RISK FACTOR, EVEN THOUGH ANIMAL STUDIES HAVE DEMONSTRATED THAT CHRONIC PATERNAL ALCOHOL CONSUMPTION HAS A DETRIMENTAL EFFECT ON THE PHYSICAL AND MENTAL DEVELOPMENT OF OFFSPRING EVEN IN THE ABSENCE OF IN UTERO ALCOHOL EXPOSURE. IT HAS BEEN DOCUMENTED THAT ALCOHOL CAN REDUCE THE LEVELS AND ACTIVITY OF DNA METHYLTRANSFERASES RESULTING IN DNA HYPOMETHYLATION AND THAT REDUCED METHYLTRANSFERASE ACTIVITY CAN CAUSE ACTIVATION OF NORMALLY SILENCED GENES. THE AIM OF THIS STUDY WAS TO ESTABLISH A LINK BETWEEN ALCOHOL USE IN MEN AND HYPOMETHYLATION OF PATERNALLY IMPRINTED LOCI IN SPERM DNA IN GENOMIC REGIONS CRITICAL FOR EMBRYONIC DEVELOPMENT, THUS PROVIDING A MECHANISM FOR PATERNAL EFFECTS IN THE AETIOLOGY OF FASD. METHODS: SPERM DNA FROM MALE VOLUNTEERS WAS BISULFITE TREATED AND THE METHYLATION PATTERNS OF 2 DIFFERENTIALLY METHYLATED REGIONS (DMRS), H19 AND IG-DMR, ANALYZED FOLLOWING SEQUENCING OF INDIVIDUAL CLONES. THE METHYLATION PATTERNS WERE CORRELATED WITH THE ALCOHOL CONSUMPTION LEVELS OF THE VOLUNTEER MALES. RESULTS: THERE WAS A PATTERN OF INCREASED DEMETHYLATION WITH ALCOHOL CONSUMPTION AT THE 2 IMPRINTED LOCI WITH A SIGNIFICANT DIFFERENCE OBSERVED AT THE IG-DMR BETWEEN THE NONDRINKING AND HEAVY ALCOHOL CONSUMING GROUPS. GREATER INTER-INDIVIDUAL VARIATION IN AVERAGE METHYLATION WAS OBSERVED AT THE H19 DMR AND INDIVIDUAL CLONES WERE MORE EXTENSIVELY DEMETHYLATED THAN THOSE OF THE IG-DMR. CPG SITE #4 IN THE IG-DMR WAS PREFERENTIALLY DEMETHYLATED AMONG ALL INDIVIDUALS AND ALONG WITH THE H19 DMR CPG SITE #7 LOCATED WITHIN THE CTCF BINDING SITE 6 SHOWED SIGNIFICANT DEMETHYLATION IN THE ALCOHOL CONSUMING GROUPS COMPARED WITH THE CONTROL GROUP. CONCLUSION: THIS STUDY DEMONSTRATES A CORRELATION BETWEEN CHRONIC ALCOHOL USE AND DEMETHYLATION OF NORMALLY HYPERMETHYLATED IMPRINTED REGIONS IN SPERM DNA. WE HYPOTHESIZE THAT, SHOULD THESE EPIGENETIC CHANGES IN IMPRINTED GENES BE TRANSMITTED THROUGH FERTILIZATION, THEY WOULD ALTER THE CRITICAL GENE EXPRESSION DOSAGES REQUIRED FOR NORMAL PRENATAL DEVELOPMENT RESULTING IN OFFSPRING WITH FEATURES OF FASD.	2009	
                                                                                                                     
13  3983  34 LONG-TERM EXPOSURE TO CIGARETTE SMOKE EXTRACT INDUCES HYPOMETHYLATION AT THE RUNX3 AND IGF2-H19 LOCI IN IMMORTALIZED HUMAN UROTHELIAL CELLS. CIGARETTE SMOKING IS THE SINGLE MOST IMPORTANT EPIDEMIOLOGICAL RISK FACTOR FOR BLADDER CANCER BUT IT IS NOT KNOWN WHETHER EXPOSURE OF UROTHELIAL CELLS TO THE SYSTEMIC SOLUBLE CONTENTS OF CIGARETTE SMOKE IS DIRECTLY CAUSATIVE TO BLADDER CANCER AND THE ASSOCIATED EPIGENETIC CHANGES SUCH AS TUMOR SUPPRESSOR GENE HYPERMETHYLATION. WE UNDERTOOK THIS STUDY TO INVESTIGATE IF LONG-TERM TREATMENT OF HUMAN UROTHELIAL CELLS WITH CIGARETTE SMOKE EXTRACT (CSE) RESULTS IN TUMOR SUPPRESSOR GENE HYPERMETHYLATION, A PHENOTYPE THAT WAS PREVIOUSLY ASSOCIATED WITH LONG-TERM CONSTANT CSE TREATMENT OF AIRWAY EPITHELIAL CELLS. WE CHRONICALLY TREATED AN IMMORTALIZED HUMAN UROTHELIAL CELL LINE UROTSA WITH CSE USING A CYCLIC DAILY REGIMEN BUT THE CELLS WERE CULTURED IN CSE-FREE MEDIUM BETWEEN DAILY TREATMENTS. BISULFITE SEQUENCING AND REAL-TIME PCR ARRAY-BASED METHYLATION PROFILING WERE EMPLOYED TO EVALUATE METHYLATION CHANGES AT TUMOR SUPPRESSOR GENE LOCI IN THE CHRONICALLY CSE-TREATED CELLS VERSUS THE PASSAGE-MATCHED UNTREATED CONTROL CELLS. THE RUNX3 TUMOR SUPPRESSOR GENE PROMOTER WAS HYPOMETHYLATED WITH A SIGNIFICANT INCREASE IN PROPORTION OF THE COMPLETELY UNMETHYLATED HAPLOTYPE AFTER THE LONG-TERM CSE TREATMENT; WHEREAS RUNX3 PROMOTER HYPERMETHYLATION WAS PREVIOUSLY REPORTED FOR BLADDER CANCERS OF SMOKERS. HYPOMETHYLATION INDUCED BY THE LONG-TERM CSE TREATMENT WAS ALSO OBSERVED FOR THE IGF2-H19 LOCUS. THE METHYLATION STATUS AT THE PRSS8/PROSTASIN AND 16 ADDITIONAL LOCI HOWEVER, WAS UNAFFECTED BY THE CHRONIC CSE TREATMENT. TRANSIENT CSE TREATMENT OVER 1 DAILY REGIMEN RESULTED IN TRANSCRIPTIONAL DOWN-REGULATION OF RUNX3 AND H19, BUT ONLY THE H19 TRANSCRIPTION WAS DOWN-REGULATED IN THE CHRONICALLY CSE-TREATED UROTHELIAL CELLS. TRANSCRIPTION OF A KEY ENZYME IN ONE-CARBON METABOLISM, DIHYDROFOLATE REDUCTASE (DHFR) WAS GREATLY REDUCED BY THE LONG-TERM CSE TREATMENT, POTENTIALLY SERVING AS A MECHANISM FOR THE HYPOMETHYLATION PHENOTYPE VIA A REDUCED SUPPLY OF METHYL DONOR. IN CONCLUSION, CHRONIC CYCLIC CSE TREATMENT OF UROTHELIAL CELLS INDUCED HYPOMETHYLATION RATHER THAN HYPERMETHYLATION AT SPECIFIC LOCI.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                         
14  4081  34 MATERNAL MICRONUTRIENT SUPPLEMENTATION SUPPRESSES T CELL CHEMOKINE RECEPTOR EXPRESSION AND FUNCTION IN F1 MICE. PRENATAL ENVIRONMENTAL EXPOSURES PLAY A CRITICAL ROLE IN DETERMINING LATE-LIFE CHRONIC DISEASE SUSCEPTIBILITY. HOWEVER, THE MECHANISMS LINKING THE IN UTERO ENVIRONMENT AND DISEASE DEVELOPMENT IN THE OFFSPRING ARE POORLY UNDERSTOOD. RECENT INVESTIGATIONS HAVE CONFIRMED A CENTRAL PATHOGENIC ROLE OF T CELL CHEMOKINE RECEPTORS, PARTICULARLY C-C CHEMOKINE RECEPTOR (CCR) 2 AND CCR5, IN CHRONIC INFLAMMATORY CONDITIONS. THIS STUDY WAS DESIGNED TO DETERMINE THE EFFECT OF A SYNTHETIC PRENATAL MICRONUTRIENT SUPPLEMENTATION (MS) DIET RICH IN METHIONINE PATHWAY METABOLITES ON THE T CELL CHEMOKINE SYSTEM IN F1 C57BL/6 MICE. FEMALE MICE WERE FED EITHER AN MS OR CONTROL DIET 3 WK PRIOR TO MATING, DURING PREGNANCY, AND LACTATION. AT 4 WK OF AGE, F1 MICE WERE KILLED FOR EXPERIMENTS OR WERE FED THE STANDARD NIH-31 DIET AND ALLOWED TO AGE. FOOD CONSUMPTION, MATERNAL WEIGHT GAIN, AND LITTER SIZE WERE SIMILAR IN DAMS FED THE CONTROL AND MS DIETS. HOWEVER, THE F1 OFFSPRING OF DAMS FED THE MS DIET WERE SMALLER IN SIZE (P < 0.001). T CELLS FROM THE MS F1 OFFSPRING HAD GLOBAL HYPERMETHYLATION COMPARED WITH CONTROL F1 OFFSPRING (P < 0.005), CORRESPONDING TO LOWER T CELL CHEMOKINE RECEPTOR EXPRESSION [CCR2 (P < 0.001), CCR5 (P < 0.001), AND C-X-C CHEMOKINE RECEPTOR 3 (P < 0.01)] AND CYTOKINE EXPRESSION [TNFALPHA (P < 0.05), IL-2 (P < 0.001), AND IL-4 (P < 0.01)]. REDUCED T CELL CHEMOKINE RECEPTOR GENE EXPRESSION IN MS F1 MICE WAS ASSOCIATED WITH DECREASED CHEMOTAXIS IN VITRO TO C-C CHEMOKINE LIGAND (CCL) 2 AND C-X-C CHEMOKINE LIGAND 10 (P < 0.01) AND IN VIVO TO CCL2 (P < 0.01). TAKEN TOGETHER, THE RESULTS SUGGEST THAT EPIGENETIC ALTERATION THROUGH PRENATAL DIET MANIPULATION REDUCES THE RESPONSE TO PROINFLAMMATORY SIGNALS IN MICE.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
15  2019  33 EPIGENETIC CHANGE IN E-CADHERIN AND COX-2 TO PREDICT CHRONIC PERIODONTITIS. BACKGROUND: DNA METHYLATION OF CERTAIN GENES FREQUENTLY OCCURS IN NEOPLASTIC CELLS. ALTHOUGH THE CAUSE REMAINS UNKNOWN, MANY GENES HAVE BEEN IDENTIFIED WITH SUCH ATYPICAL METHYLATION IN NEOPLASTIC CELLS. THE HYPERMETHYLATION OF E-CADHERIN AND CYCLOOXYGENASE 2 (COX-2) IN CHRONIC INFLAMMATION SUCH AS CHRONIC PERIODONTITIS MAY DEMONSTRATE MILD LESION/MUTATION EPIGENETIC LEVEL. THIS STUDY COMPARES THE HYPERMETHYLATION STATUS OF E-CADHERIN AND COX-2 GENES WHICH ARE OFTEN FOUND IN BREAST CANCER PATIENTS WITH THAT IN CHRONIC PERIODONTITIS. METHODS: TOTAL DNA WAS EXTRACTED FROM THE BLOOD SAMPLES OF 108 SYSTEMICALLY HEALTHY NON-PERIODONTITIS SUBJECTS, AND THE GINGIVAL TISSUES AND BLOOD SAMPLES OF 110 CHRONIC PERIODONTITIS PATIENT AS WELL AS NEOPLASTIC TISSUES OF 106 BREAST CANCER PATIENTS. METHYLATION-SPECIFIC PCR FOR E-CADHERIN AND COX-2 WAS PERFORMED ON THESE SAMPLES AND THE PCR PRODUCTS WERE ANALYZED ON 2% AGAROSE GEL. RESULTS: HYPERMETHYLATION OF E-CADHERIN AND COX-2 WAS OBSERVED IN 38% AND 35% OF THE BREAST CANCER SAMPLES, RESPECTIVELY. IN CHRONIC PERIODONTITIS PATIENTS THE DETECTION RATE WAS 25% AND 19% RESPECTIVELY, AND NONE WAS FOUND IN THE SYSTEMICALLY HEALTHY NON-PERIODONTITIS CONTROL SUBJECTS. THE HYPERMETHYLATION STATUS WAS SHOWN TO BE CORRELATED AMONG THE THREE GROUPS WITH STATISTICAL SIGNIFICANCE (P < 0.0001). THE METHYLATION OF CPG ISLANDS IN E-CADHERIN AND COX-2 GENES IN PERIODONTITIS PATIENTS OCCURS MORE FREQUENTLY IN PERIODONTITIS PATIENTS THAN IN THE CONTROL SUBJECTS, BUT OCCURS LESS FREQUENTLY THAN IN THE BREAST CANCER PATIENTS. CONCLUSIONS: THIS SET OF DATA SHOWS THAT THE EPIGENETIC CHANGE IN E-CADHERIN AND CYCLOOXYGENASE-2 IS ASSOCIATED WITH CHRONIC PERIODONTITIS. THE EPIGENETIC CHANGES PRESENTED IN CHRONIC INFLAMMATION PATIENTS MIGHT DEMONSTRATE AN IRREVERSIBLE DESTRUCTION IN THE TISSUES OR ORGANS SIMILAR TO THE EFFECTS OF CANCER. CHRONIC PERIODONTITIS TO SOME EXTENT MIGHT BE ASSOCIATED WITH DNA HYPERMETHYLATION WHICH IS RELATED TO CANCER RISK FACTORS.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
16  3301  28 HIGH-FAT OR ETHINYL-OESTRADIOL INTAKE DURING PREGNANCY INCREASES MAMMARY CANCER RISK IN SEVERAL GENERATIONS OF OFFSPRING. MATERNAL EXPOSURES TO ENVIRONMENTAL FACTORS DURING PREGNANCY INFLUENCE THE RISK OF MANY CHRONIC ADULT-ONSET DISEASES IN THE OFFSPRING. HERE WE INVESTIGATE WHETHER FEEDING PREGNANT RATS A HIGH-FAT (HF)- OR ETHINYL-OESTRADIOL (EE2)-SUPPLEMENTED DIET AFFECTS CARCINOGEN-INDUCED MAMMARY CANCER RISK IN DAUGHTERS, GRANDDAUGHTERS AND GREAT-GRANDDAUGHTERS. WE SHOW THAT MAMMARY TUMOURIGENESIS IS HIGHER IN DAUGHTERS AND GRANDDAUGHTERS OF HF RAT DAMS AND IN DAUGHTERS AND GREAT-GRANDDAUGHTERS OF EE2 RAT DAMS. OUTCROSS EXPERIMENTS SUGGEST THAT THE INCREASE IN MAMMARY CANCER RISK IS TRANSMITTED TO HF GRANDDAUGHTERS EQUALLY THROUGH THE FEMALE OR MALE GERM LINES, BUT IT IS ONLY TRANSMITTED TO EE2 GRANDDAUGHTERS THROUGH THE FEMALE GERM LINE. THE EFFECTS OF MATERNAL EE2 EXPOSURE ON OFFSPRING'S MAMMARY CANCER RISK ARE ASSOCIATED WITH CHANGES IN THE DNA METHYLATION MACHINERY AND METHYLATION PATTERNS IN MAMMARY TISSUE OF ALL THREE EE2 GENERATIONS. WE CONCLUDE THAT DIETARY AND OESTROGENIC EXPOSURES IN PREGNANCY INCREASE BREAST CANCER RISK IN MULTIPLE GENERATIONS OF OFFSPRING, POSSIBLY THROUGH EPIGENETIC MEANS.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
17  4008  34 LOW DOSE OF URANIUM INDUCES MULTIGENERATIONAL EPIGENETIC EFFECTS IN RAT KIDNEY. PURPOSE: A PROTOCOL OF CHRONIC EXPOSURE TO LOW DOSE OF URANIUM WAS ESTABLISHED IN ORDER TO DISTINGUISH THE SEXUAL DIFFERENCES AND THE DEVELOPMENTAL PROCESS THAT ARE CRITICAL WINDOWS FOR EPIGENETIC EFFECTS OVER GENERATIONS. METHODS: BOTH MALE AND FEMALE RATS WERE CONTAMINATED THROUGH THEIR DRINKING WATER WITH A NON-TOXIC SOLUTION OF URANYL NITRATE FOR 9 MONTHS. THE EXPOSED GENERATION (F0) AND THE FOLLOWING TWO GENERATIONS (F1 AND F2) WERE EXAMINED. CLINICAL MONITORING, GLOBAL DNA METHYLATION PROFILE AND DNA METHYLTRANSFERASES (DNMTS) GENE EXPRESSION WERE ANALYZED IN KIDNEYS. RESULTS: WHILE THE BODY WEIGHT OF F1 MALES INCREASED, A SMALL DECREASE IN KIDNEY AND BODY WEIGHT WAS OBSERVED IN F2 MALES. IN ADDITION, GLOBAL DNA HYPERMETHYLATION PROFILE IN KIDNEY CELLS WAS OBSERVED IN F1 AND F2 MALES. QPCR RESULTS REVEAL A SIGNIFICANT INCREASE OF METHYLTRANSFERASE GENES EXPRESSION (DNMT1 AND DNMT3A) FOR F2 FEMALES. CONCLUSIONS: IN THE FIELD OF PUBLIC HEALTH POLICY AND TO RAISE ATTENTION TO GENERATIONAL EFFECTS FOR THE RISK ASSESSMENT OF THE ENVIRONMENTAL EXPOSURES, LOW DOSES OF URANIUM DO NOT IMPLY CLINICAL EFFECTS ON ADULT EXPOSED RATS. HOWEVER, OUR RESULTS CONFIRM THE IMPORTANCE OF THE DEVELOPMENTAL WINDOWS' SENSITIVITY IN ADDITION TO THE SEXUAL DIMORPHISMS OF THE OFFSPRING.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
18  2436  39 EPIGENETIC SILENCING OF SOCS3 IDENTIFIES A SUBSET OF PROSTATE CANCER WITH AN AGGRESSIVE BEHAVIOR. BACKGROUND: CHRONIC INFLAMMATION AND SUBSEQUENT TISSUTAL ALTERATIONS MAY PLAY A KEY ROLE IN PROSTATE CARCINOGENESIS. IN THIS WAY, MOLECULAR ALTERATIONS OF THE SUPPRESSOR OF CYTOKINE SIGNALING 3 (SOCS3), ONE OF THE MOST IMPORTANT INHIBITORY MOLECULE OF INFLAMMATORY SIGNAL TRANSDUCTION CIRCUITRIES, COULD CONTRIBUTE TO EXPLAIN THE PLEIOTROPIC ROLE OF INTERLEUKIN-6 (IL-6) IN THIS TYPE OF CANCER. METHODS: WE ANALYZED THE METHYLATION STATUS AND MRNA EXPRESSION OF SOCS3 IN 20 BENIGN PROSTATE HYPERPLASIAS (BPH) AND IN 51 PROSTATE CANCER SPECIMENS. WE ANALYZED THE SOCS3 METHYLATION STATUS USING METHYLATION-SPECIFIC PCR. HYPERMETHYLATION WAS CONFIRMED BY SEQUENCING AFTER SUBCLONING. EPIGENETIC SILENCING OF THIS GENE WAS ALSO DEMONSTRATED BY REAL-TIME PCR AND BY IMMUNOHISTOCHEMISTRY. RESULTS AND CORRELATION WITH CLINICAL DATA WERE STATISTICALLY ANALYZED. RESULTS: WE FOUND THAT THE PROMOTER OF SOCS3 WAS METHYLATED IN 39.2% OF PROSTATE CANCER. ON THE CONTRARY, ALL BPH AND NORMAL CONTROLS HAD AN UNMETHYLATED PATTERN. REAL-TIME ANALYSIS SHOWED THAT IN METHYLATED CASES SOCS3 MRNA EXPRESSION WAS REDUCED BY THREE AND FOUR FOLDS AS COMPARED TO BPH AND UNMETHYLATED CASES, RESPECTIVELY. INTERESTINGLY, SOCS3 MRNA LEVEL WAS HIGHER IN UNMETHYLATED PROSTATE CANCER THAN IN BPH. THE IMMUNOHISTOCHEMICAL STAINING ANALYSIS FOR SOCS 3 CONFIRMED MRNA RESULTS. MOREOVER, METHYLATION OF SOCS3 PROMOTER SIGNIFICANTLY ASSOCIATED WITH INTERMEDIATE-HIGH GRADE GLEASON SCORE (P = 0.0007) AND WITH AN UNFAVORABLE CLINICAL OUTCOME (P = 0.0019). CONCLUSIONS: OUR DATA SUGGEST THAT SOCS3 HYPERMETHYLATION MAY BE INVOLVED IN THE PATHOGENESIS OF PROSTATE CANCER AND COULD IDENTIFY A TUMOR SUBSET WITH AN AGGRESSIVE BEHAVIOR.	2011	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
19  3646  36 INCREASED PROTEIN EXPRESSION OF DNA METHYLTRANSFERASE (DNMT) 1 IS SIGNIFICANTLY CORRELATED WITH THE MALIGNANT POTENTIAL AND POOR PROGNOSIS OF HUMAN HEPATOCELLULAR CARCINOMAS. ALTERATION OF DNA METHYLATION IS ONE OF THE MOST CONSISTENT EPIGENETIC CHANGES IN HUMAN CANCERS. DNA METHYLTRANSFERASE (DNMT) 1 IS A MAJOR ENZYME INVOLVED IN ESTABLISHING GENOMIC METHYLATION PATTERNS. MOST OF THE STUDIES CONCERNING DNMT1 EXPRESSION IN HUMAN CANCERS HAVE BEEN PERFORMED ONLY AT THE MRNA LEVEL. TO DIRECTLY EXAMINE DNMT1 PROTEIN EXPRESSION LEVELS DURING HUMAN HEPATOCARCINOGENESIS, 16 HISTOLOGICALLY NORMAL LIVER TISSUES, 51 NONCANCEROUS LIVER TISSUES EXHIBITING CHRONIC HEPATITIS OR CIRRHOSIS, WHICH ARE CONSIDERED TO BE PRECANCEROUS CONDITIONS, AND 53 HEPATOCELLULAR CARCINOMAS (HCCS) WERE SUBJECTED TO IMMUNOHISTOCHEMIC EXAMINATION. IF MORE THAN 20% OF THE CELLS EXHIBITED NUCLEAR DNMT1 STAINING, THE TISSUE SAMPLE WAS CONSIDERED TO BE DNMT1-POSITIVE. DNMT1 IMMUNOREACTIVITY WAS OBSERVED IN 23 (43%) OF THE HCCS, BUT IN NONE (0%) OF THE HISTOLOGICALLY NORMAL LIVER OR NONCANCEROUS LIVER TISSUES EXHIBITING CHRONIC HEPATITIS OR CIRRHOSIS. THE INCIDENCE OF INCREASED DNMT1 PROTEIN EXPRESSION IN HCCS CORRELATED SIGNIFICANTLY WITH POOR TUMOR DIFFERENTIATION (P = 0.0006) AND PORTAL VEIN INVOLVEMENT (P = 0.0002). MOREOVER, THE RECURRENCE-FREE (P = 0.0001) AND OVERALL (P < 0.0001) SURVIVAL RATES OF PATIENTS WITH HCCS EXHIBITING INCREASED DNMT1 PROTEIN EXPRESSION WERE SIGNIFICANTLY LOWER THAN THOSE OF PATIENTS WITH HCCS THAT DID NOT EXHIBIT INCREASED EXPRESSION. INCREASED DNMT1 PROTEIN EXPRESSION MAY PLAY A CRITICAL ROLE IN THE MALIGNANT PROGRESSION OF HCCS AND BE A BIOLOGIC PREDICTOR OF BOTH HCC RECURRENCE AND A POOR PROGNOSIS IN HCC PATIENTS.	2003	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
20  3460  32 HYPOMETHYLATION OF THE IL8 PROMOTER IN NASAL EPITHELIAL CELLS OF PATIENTS WITH CHRONIC RHINOSINUSITIS WITH NASAL POLYPS. BACKGROUND: IL-8 IS AN IMPORTANT CHEMOKINE IMPLICATED IN THE PATHOGENESIS OF CHRONIC RHINOSINUSITIS (CRS), BUT LITTLE IS KNOWN ABOUT EPIGENETIC REGULATION OF IL8 IN THE PATHOGENESIS OF CRS. OBJECTIVE: WE SOUGHT TO INVESTIGATE THE RELATIONSHIP BETWEEN THE DNA METHYLATION LEVEL IN THE IL8 PROXIMAL PROMOTER AND CRS IN HAN CHINESE SUBJECTS. METHODS: PATIENTS WITH CHRONIC RHINOSINUSITIS WITH NASAL POLYPS (CRSWNP; N = 187), PATIENTS WITH CHRONIC RHINOSINUSITIS WITHOUT NASAL POLYPS (CRSSNP; N = 89), AND CONTROL SUBJECTS (N = 57) WERE ENROLLED IN 2 INDEPENDENT COHORTS. PURIFIED HUMAN NASAL EPITHELIAL CELLS FROM EACH PARTICIPANT WERE ASSESSED FOR PERCENTAGE DNA METHYLATION OF CPG SITES IN THE IL8 PROXIMAL PROMOTER BY USING BISULFITE PYROSEQUENCING AND FOR FUNCTIONAL ASPECTS OF METHYLATION STATUS BY USING IN VITRO ASSAYS. RESULTS: DNA METHYLATION OF CPG SITES 1, 2, AND 3, RESPECTIVELY, IN THE IL8 PROXIMAL PROMOTER WAS SIGNIFICANTLY DECREASED IN HUMAN NASAL EPITHELIAL CELLS OF PATIENTS WITH CRSWNP COMPARED WITH THAT IN PATIENTS WITH CRSSNP (P < .001) AND CONTROL SUBJECTS (P < .001). PERCENTAGE OF DNA METHYLATION OF THE CPG3 SITE WAS CORRELATED NEGATIVELY WITH BOTH TISSUE EOSINOPHILIC CATIONIC PROTEIN (P < .01) AND MYELOPEROXIDASE (P < .05) LEVELS. IL-1BETA (P < .001) AND TNF-ALPHA (P < .01) SIGNIFICANTLY INCREASED IL8 EXPRESSION ACCOMPANIED BY A REDUCTION IN METHYLATION AT THE CPG3 SITE (P < .001). ELECTROPHORETIC MOBILITY SHIFT ASSAYS DEMONSTRATED THAT METHYLATION STATUS OF CPG3 CHANGED THE BINDING OF OCTAMER-BINDING TRANSCRIPTION FACTOR 1 AND NUCLEAR FACTOR KAPPAB. CONCLUSION: DECREASED DNA METHYLATION OF PARTICULARLY CPG SITES IN THE IL8 PROXIMAL PROMOTER MIGHT PLAY A ROLE IN THE PATHOGENESIS OF CRSWNP.	2019