1 1596 157 DNA METHYLATION REDUCES THE YES-ASSOCIATED PROTEIN 1/WW DOMAIN CONTAINING TRANSCRIPTION REGULATOR 1 PATHWAY AND PREVENTS PATHOLOGIC REMODELING DURING BLADDER OBSTRUCTION BY LIMITING EXPRESSION OF BDNF. CHRONIC BLADDER OBSTRUCTION AND BLADDER SMOOTH MUSCLE CELL (SMC) STRETCH PROVIDE FIBROTIC AND MECHANICAL ENVIRONMENTS THAT CAN LEAD TO EPIGENETIC CHANGE. THEREFORE, WE EXAMINED THE ROLE OF DNA METHYLATION IN BLADDER PATHOLOGY AND TRANSCRIPTIONAL CONTROL. SPRAGUE-DAWLEY FEMALE RATS UNDERWENT PARTIAL BLADDER OBSTRUCTION BY LIGATION OF A SILK SUTURE AROUND THE PROXIMAL URETHRA NEXT TO A 0.9-MM STEEL ROD. SHAM OPERATION COMPRISED PASSING THE SUTURE AROUND THE URETHRA. AFTER 2 WEEKS, RATS WERE RANDOMIZED TO NORMAL SALINE OR DNA METHYLTRANSFERASE INHIBITOR, 5-AZA-2-DEOXYCYTIDINE (DAC) AT 1 MG/KG, THREE TIMES/WEEK INTRAPERITONEALLY. AFTER 6 WEEKS, BLADDERS WERE WEIGHED AND DIVIDED FOR HISTOLOGY AND RNA ANALYSIS BY HIGH-THROUGHPUT REAL-TIME QUANTITATIVE PCR ARRAYS. DAC TREATMENT DURING OBSTRUCTION IN VIVO PROFOUNDLY AUGMENTED BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) EXPRESSION COMPARED WITH THE OBSTRUCTION WITH VEHICLE GROUP, WHICH WAS STATISTICALLY CORRELATED WITH PATHOPHYSIOLOGIC PARAMETERS. BDNF, CYSTEINE RICH ANGIOGENIC INDUCER 61 (CYR61), AND CONNECTIVE TISSUE GROWTH FACTOR (CTGF) EXPRESSION CLUSTERED TIGHTLY TOGETHER USING PEARSON'S CORRELATION ANALYSIS. THEIR PROMOTERS WERE ASSOCIATED WITH THE TEA DOMAIN FAMILY MEMBER 1 (TEAD1) AND YES-ASSOCIATED PROTEIN 1/WW DOMAIN CONTAINING TRANSCRIPTION REGULATOR 1 PATHWAYS. INTERESTINGLY, DAC TREATMENT INCREASED BDNF EXPRESSION IN BLADDER SMCS (P < 0.0002). STRETCH-INDUCED BDNF WAS INHIBITED BY THE YAP/WWTR1 INHIBITOR VERTEPORFIN. VERTEPORFIN IMPROVED THE SMC PHENOTYPE (PROLIFERATIVE MARKERS AND SMC MARKER EXPRESSION), IN PART BY REDUCING BDNF. EXPRESSION OF BDNF IS LIMITED BY DNA METHYLATION AND ASSOCIATED WITH PATHOPHYSIOLOGIC CHANGES DURING PARTIAL BLADDER OUTLET OBSTRUCTION AND SMC PHENOTYPIC CHANGE IN VITRO.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
2 2779  52 EZH2 AND MATRIX CO-REGULATE PHENOTYPE AND KCNB2 EXPRESSION IN BLADDER SMOOTH MUSCLE CELLS. BACKGROUND: PARTIAL BLADDER OUTLET OBSTRUCTION (PBO) IS A WIDESPREAD CAUSE OF URINARY DYSFUNCTION AND PATIENT DISCOMFORT, RESULTING IN IMMENSE HEALTH CARE COSTS. PREVIOUSLY, WE FOUND THAT OBSTRUCTION IS ASSOCIATED WITH ALTERED REGULATION OF EPIGENETIC MACHINERY AND ALTERED FUNCTION. HERE WE EXAMINED IF PBO AND CHRONIC BLADDER OBSTRUCTIVE DISEASE (COBD) AFFECT EPIGENETIC MARKS IN A PROOF OF PRINCIPLE GENE AND EXPLORED MECHANISMS OF ITS EPIGENETIC REGULATION USING IN VITRO MODELS. METHODS: ARCHIVAL OBSTRUCTION TISSUES FROM COBD HAD BEEN CREATED IN 200-250 G FEMALE SPRAGUE-DAWLEY RATS BY SURGICAL LIGATION OF THE URETHRA FOR 6 WEEKS, FOLLOWED BY REMOVAL OF THE SUTURE AND FOLLOWING ANIMALS FOR 6 MORE WEEKS. OBSTRUCTION (PBO) IS THE 6-WEEK LIGATION ONLY. SHAM LIGATIONS COMPRISE PASSING THE SUTURE BEHIND THE URETHRA. HISTONE3 LYSINE27 TRIMETHYLATION (H3K27ME3) WAS STUDIED BY IMMUNOSTAINING AND CHROMATIN IMMUNOPRECIPITATION (CHIP)/PCR. THE INTERACTION OF MATRIX WITH KCNB2 REGULATION WAS STUDIED IN HUMAN BLADDER SMC PLATED ON DAMAGED MATRIX AND NATIVE COLLAGEN AND TREATED WITH VEHICLE OR UNC1999. CELLS WERE ANALYZED BY IMMUNOSTAINING FOR CELL PHENOTYPE, AND WESTERN BLOTTING FOR KCNB2, H3K27ME3 AND EZH2. EFFECTS OF CONDITIONED MEDIA FROM THESE CELLS WERE ALSO EXAMINED ON CELL PHENOTYPE. SIRNA AGAINST KCNB2 WAS EXAMINED FOR EFFECTS ON CELL PHENOTYPE AND GENE EXPRESSION BY RT-QPCR. RESULTS: H3K27ME3 INCREASED BY IMMUNOFLUORESCENCE DURING PBO, AND BY CHIP/PCR DURING COBD IN THE CPG ISLAND (CGI) AS WELL AS 350 BP UPSTREAM. OBSTRUCTION VS. SHAM ALSO SHOWED AN INCREASE IN H3K27ME3 DEPOSITION. IN SMC IN VITRO, EZH2 INHIBITION RESTORED KCNB2 EXPRESSION AND PARTIALLY RESTORED SMC PHENOTYPE. CONCLUSIONS: REGULATION OF KCNB2 AT THE PROMOTER DEMONSTRATED DYNAMIC CHANGES IN H3K27ME3 DURING COBD AND OBSTRUCTION. IN VITRO MODELS SUGGEST THAT MATRIX PLAYS A ROLE IN REGULATION OF EZH2, H3K27ME3 AND KCNB2, WHICH MAY PLAY A ROLE IN THE REGULATION OF SMOOTH MUSCLE PHENOTYPE IN VIVO.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
3 3722  70 INHIBITION OF DNA METHYLATION DURING CHRONIC OBSTRUCTIVE BLADDER DISEASE (COBD) IMPROVES FUNCTION, PATHOLOGY AND EXPRESSION. PARTIAL BLADDER OUTLET OBSTRUCTION DUE TO PROSTATE HYPERPLASIA OR POSTERIOR URETHRAL VALVES, IS A WIDESPREAD CAUSE OF URINARY DYSFUNCTION, PATIENT DISCOMFORT AND ALSO RESPONSIBLE FOR IMMENSE HEALTH CARE COSTS. EVEN AFTER REMOVAL OR RELIEF OF OBSTRUCTION, THE FUNCTIONAL AND PATHOLOGIC ASPECTS OF OBSTRUCTION REMAIN AS A CHRONIC OBSTRUCTIVE BLADDER DISEASE (COBD). EPIGENETIC CHANGES, SUCH AS DNA METHYLATION, CONTRIBUTE TO THE PERSISTENT CHARACTER OF MANY CHRONIC DISEASES, AND MAY BE ALTERED IN COBD. WE TESTED WHETHER CANDIDATE GENES AND PATHWAYS AND THE PATHOPHYSIOLOGY OF COBD WERE AFFECTED BY A HYPOMETHYLATING AGENT, DECITABINE (DAC). COBD WAS CREATED IN FEMALE SPRAGUE-DAWLEY RATS BY SURGICAL LIGATION OF THE URETHRA FOR 6 WEEKS, FOLLOWED BY REMOVAL OF THE SUTURE. SHAM LIGATIONS WERE PERFORMED BY PASSING THE SUTURE BEHIND THE URETHRA. AFTER REMOVAL OF THE OBSTRUCTION OR SHAM REMOVAL, ANIMALS WERE RANDOMIZED TO DAC TREATMENT (1 MG/KG/3-TIMES/WEEK INTRAPERITONEALLY) OR VEHICLE (NORMAL SALINE). BLADDER FUNCTION WAS NON-INVASIVELY TESTED USING METABOLIC CAGES, BOTH ONE DAY PRIOR TO DE-OBSTRUCTION AT 6 WEEKS AND PRIOR TO SACRIFICE AT 10 WEEKS. RESIDUAL VOLUME AND BLADDER MASS WERE MEASURED FOR EACH BLADDER. BLADDERS WERE EXAMINED BY IMMUNOSTAINING AS WELL AS QPCR. THE EFFECTS OF DNA METHYLTRANSFERASE (DNMT)-3A KNOCKOUT OR OVEREXPRESSION ON SMOOTH MUSCLE CELL (SMC) FUNCTION AND PHENOTYPE WERE ALSO EXAMINED IN BLADDER SMC AND EX VIVO CULTURE. RESIDUAL VOLUMES OF THE DAC TREATED GROUP WERE NOT SIGNIFICANTLY DIFFERENT FROM THE NS GROUP. COMPARED TO COBD NS, COBD DAC TREATMENT HELPED PRESERVE MICTURITION VOLUME WITH A SIGNIFICANT RECOVERY OF THE VOIDING EFFICIENCY (RATIO OF THE MAXIMUM VOIDED VOLUME/MAXIMUM BLADDER CAPACITY) BY ONE THIRD (FIG. 1, P > 0.05). BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) VARIANTS 1 AND 5 WERE UPREGULATED BY COBD AND SIGNIFICANTLY REDUCED BY DAC TREATMENT. DEPOSITION OF COLLAGEN IN THE COBD BLADDER WAS REDUCED BY DAC, BUT GROSS HYPERTROPHY REMAINED. IN BLADDER SMC, DNMT3A OVEREXPRESSION LED TO A LOSS OF CONTRACTILE FUNCTION AND PHENOTYPE. IN BLADDERS, PERSISTENTLY ALTERED BY COBD, INHIBITION OF DNA-METHYLATION ENHANCES FUNCTIONAL RECOVERY, UNLIKE TREATMENT DURING PARTIAL OBSTRUCTION, WHICH EXACERBATES OBSTRUCTIVE PATHOLOGY. THE UNDERLYING MECHANISMS MAY RELATE TO THE GENE EXPRESSION CHANGES IN BDNF AND THEIR EFFECTS ON SIGNALING IN THE BLADDER.	2021	
                 
4 2774  36 EXTRACELLULAR SUPEROXIDE DISMUTASE (EC-SOD) REGULATES GENE METHYLATION AND CARDIAC FIBROSIS DURING CHRONIC HYPOXIC STRESS. CHRONIC HYPOXIC STRESS INDUCES EPIGENETIC MODIFICATIONS MAINLY DNA METHYLATION IN CARDIAC FIBROBLASTS, INACTIVATING TUMOR SUPPRESSOR GENES (RASSF1A) AND ACTIVATING KINASES (ERK1/2) LEADING TO FIBROBLAST PROLIFERATION AND CARDIAC FIBROSIS. THE RAS/ERK SIGNALING PATHWAY IS AN INTRACELLULAR SIGNAL TRANSDUCTION CRITICALLY INVOLVED IN FIBROBLAST PROLIFERATION. RASSF1A FUNCTIONS THROUGH ITS EFFECT ON DOWNSTREAM ERK1/2. THE ANTIOXIDANT ENZYME, EXTRACELLULAR SUPEROXIDE DISMUTASE (EC-SOD), DECREASES OXIDATIVE STRESS FROM CHRONIC HYPOXIA, BUT ITS EFFECTS ON THESE EPIGENETIC CHANGES HAVE NOT BEEN FULLY EXPLORED. TO TEST OUR HYPOTHESIS, WE USED AN IN-VITRO MODEL: WILD-TYPE C57B6 MALE MICE (WT) AND TRANSGENIC MALES WITH AN EXTRA COPY OF HUMAN HEC-SOD (TG). THE STUDIED ANIMALS WERE HOUSED IN HYPOXIA (10% O(2)) FOR 21 DAYS. THE RIGHT VENTRICULAR TISSUE WAS STUDIED FOR CARDIAC FIBROSIS MARKERS USING RT-PCR AND WESTERN BLOT ANALYSES. PRIMARY C57BL6 MOUSE CARDIAC FIBROBLAST TISSUE CULTURE WAS USED TO STUDY THE IN-VITRO MODEL, THE DOWNSTREAM EFFECTS OF RASSF-1 EXPRESSION AND METHYLATION, AND ITS RELATION TO ERK1/2. OUR FINDINGS SHOWED A SIGNIFICANT INCREASE IN CARDIAC FIBROSIS MARKERS: COLLAGEN 1, ALPHA SMOOTH MUSCLE ACTIN (ASMA), AND SNAIL, IN THE WT HYPOXIC ANIMALS AS COMPARED TO THE TG HYPOXIC GROUP (P < 0.05). THE EXPRESSION OF DNA METHYLATION ENZYMES (DNMT 1&3B) WAS SIGNIFICANTLY INCREASED IN THE WT HYPOXIC MICE AS COMPARED TO THE HYPOXIC TG MICE (P < 0.001). RASSF1A EXPRESSION WAS SIGNIFICANTLY LOWER AND ERK1/2 WAS SIGNIFICANTLY HIGHER IN HYPOXIA WT COMPARED TO THE HYPOXIC TG GROUP (P < 0.05). USE OF SIRNA TO BLOCK RASSF1A GENE EXPRESSION IN MURINE CARDIAC FIBROBLAST TISSUE CULTURE LED TO INCREASED FIBROBLAST PROLIFERATION (P < 0.05). METHYLATION OF THE RASSF1A PROMOTER REGION WAS SIGNIFICANTLY REDUCED IN THE TG HYPOXIC GROUP COMPARED TO THE WT HYPOXIC GROUP (0.59 VS. 0.75, RESPECTIVELY). BASED ON OUR FINDINGS, WE CAN SPECULATE THAT EC-SOD SIGNIFICANTLY ATTENUATES RASSF1A GENE METHYLATION AND CAN ALLEVIATE CARDIAC FIBROSIS INDUCED BY HYPOXIA.	2021	
                                                                                                                                                                                                                                                                                                                                                                 
5 6411  36 THE SITE SPECIFIC DEMETHYLATION IN THE 5'-REGULATORY AREA OF NMDA RECEPTOR 2B SUBUNIT GENE ASSOCIATED WITH CIE-INDUCED UP-REGULATION OF TRANSCRIPTION. BACKGROUND: THE NMDA RECEPTOR REPRESENTS A PARTICULARLY IMPORTANT SITE OF ETHANOL ACTION IN THE CNS. WE RECENTLY REPORTED THAT NMDA RECEPTOR 2B (NR2B) GENE EXPRESSION WAS PERSISTENTLY UP-REGULATED FOLLOWING CHRONIC INTERMITTENT ETHANOL (CIE) TREATMENT. INCREASING EVIDENCE THAT EPIGENETIC MECHANISMS ARE INVOLVED IN DYNAMIC AND LONG-LASTING REGULATION OF GENE EXPRESSION IN MULTIPLE NEUROADAPTIVE PROCESSES PROMPTED US TO INVESTIGATE THE ROLE OF DNA METHYLATION IN MEDIATING CIE-INDUCED UP-REGULATION OF NR2B GENE TRANSCRIPTION. TO DISSECT THE CHANGES OF DNA METHYLATION IN THE NR2B GENE, WE HAVE SCREENED A LARGE NUMBER OF CPG SITES WITHIN ITS 5'-REGULATORY AREA FOLLOWING CIE TREATMENT. METHODS: PRIMARY CORTICAL CULTURED NEURONS WERE SUBJECTED TO ETHANOL TREATMENT IN A CIE PARADIGM. BISULFITE CONVERSION FOLLOWED BY PYROSEQUENCING WAS USED FOR QUANTITATIVE MEASUREMENT AND ANALYSIS OF CPG METHYLATION STATUS WITHIN THE 5'-REGULATORY AREA OF THE NR2B GENE; CHROMATIN IMMUNOPRECIPITATION (CHIP) ASSAY WAS USED TO EXAMINE DNA LEVELS ASSOCIATED WITH METHYLATION AND TRANSCRIPTION FACTOR BINDING. ELECTROPHORETIC MOBILITY SHIFT ASSAY (EMSA) AND IN VITRO DNA METHYLATION ASSAYS WERE PERFORMED TO DETERMINE THE DIRECT IMPACT OF DNA METHYLATION ON THE INTERACTION BETWEEN DNA AND TRANSCRIPTION FACTOR AND PROMOTER ACTIVITY. RESULTS: ANALYSIS OF INDIVIDUAL CPG METHYLATION SITES WITHIN THE NR2B 5'REGULATORY AREA REVEALED THREE REGIONS WITH CLUSTERS OF SITE-SPECIFIC CPG DEMETHYLATION FOLLOWING CIE TREATMENT AND WITHDRAWAL. THIS WAS CONFIRMED BY CHIP SHOWING SIMILAR DECREASES OF METHYLATED DNA IN THE SAME REGIONS. THE CIE-INDUCED DEMETHYLATION IS CHARACTERIZED BY BEING LOCATED NEAR CERTAIN TRANSCRIPTION FACTOR BINDING SEQUENCES, AP-1 AND CRE, AND OCCURRED DURING TREATMENT AS WELL AS AFTER ETHANOL WITHDRAWAL. FURTHERMORE, THE INCREASE IN VITRO OF METHYLATED DNA DECREASED TRANSCRIPTION FACTOR BINDING ACTIVITY AND PROMOTER ACTIVITY. AN ADDITIONAL CHIP ASSAY INDICATED THAT THE CIE-INDUCED DNA DEMETHYLATION IS ACCOMPANIED BY INCREASED OCCUPATION BY TRANSCRIPTION FACTORS. CONCLUSIONS: THESE RESULTS SUGGEST AN IMPORTANT ROLE OF DNA DEMETHYLATION IN MEDIATING CIE-INDUCED NR2B GENE UP-REGULATION, THUS IMPLICATING A NOVEL MOLECULAR SITE OF ALCOHOL ACTION.	2010	
                                                                                                                                       
6  164  33 ABNORMAL HISTONE METHYLATION IS RESPONSIBLE FOR INCREASED VASCULAR ENDOTHELIAL GROWTH FACTOR 165A SECRETION FROM AIRWAY SMOOTH MUSCLE CELLS IN ASTHMA. VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF), A KEY ANGIOGENIC MOLECULE, IS ABERRANTLY EXPRESSED IN SEVERAL DISEASES INCLUDING ASTHMA WHERE IT CONTRIBUTES TO BRONCHIAL VASCULAR REMODELING AND CHRONIC INFLAMMATION. ASTHMATIC HUMAN AIRWAY SMOOTH MUSCLE CELLS HYPERSECRETE VEGF, BUT THE MECHANISM IS UNCLEAR. IN THIS STUDY, WE DEFINED THE MECHANISM IN HUMAN AIRWAY SMOOTH MUSCLE CELLS FROM NONASTHMATIC AND ASTHMATIC PATIENTS. WE FOUND THAT ASTHMATIC CELLS LACKED A REPRESSION COMPLEX AT THE VEGF PROMOTER, WHICH WAS PRESENT IN NONASTHMATIC CELLS. RECRUITMENT OF G9A, TRIMETHYLATION OF HISTONE H3 AT LYSINE 9 (H3K9ME3), AND A RESULTANT DECREASE IN RNA POLYMERASE II AT THE VEGF PROMOTER WAS CRITICAL TO REPRESSION OF VEGF SECRETION IN NONASTHMATIC CELLS. AT THE ASTHMATIC PROMOTER, H3K9ME3 WAS ABSENT BECAUSE OF FAILED RECRUITMENT OF G9A; RNA POLYMERASE II BINDING, IN ASSOCIATION WITH TATA-BINDING PROTEIN-ASSOCIATED FACTOR 1, WAS INCREASED; H3K4ME3 WAS PRESENT; AND SP1 BINDING WAS EXAGGERATED AND SUSTAINED. IN CONTRAST, DNA METHYLATION AND HISTONE ACETYLATION WERE SIMILAR IN ASTHMATIC AND NONASTHMATIC CELLS. THIS IS THE FIRST STUDY, TO OUR KNOWLEDGE, TO SHOW THAT AIRWAY CELLS IN ASTHMA HAVE ALTERED EPIGENETIC REGULATION OF REMODELING GENE(S). HISTONE METHYLATION AT GENES SUCH AS VEGF MAY BE AN IMPORTANT NEW THERAPEUTIC TARGET.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
7 1144  39 CONCOMITANT HETEROCHROMATINISATION AND DOWN-REGULATION OF GENE EXPRESSION UNVEILS EPIGENETIC SILENCING OF RELB IN AN AGGRESSIVE SUBSET OF CHRONIC LYMPHOCYTIC LEUKEMIA IN MALES. BACKGROUND: THE SENSITIVITY OF CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) CELLS TO CURRENT TREATMENTS, BOTH IN VITRO AND IN VIVO, RELIES ON THEIR ABILITY TO ACTIVATE APOPTOTIC DEATH. CLL CELLS RESISTANT TO DNA DAMAGE-INDUCED APOPTOSIS DISPLAY DEREGULATION OF A SPECIFIC SET OF GENES. METHODS: MICROARRAY HYBRIDIZATION (HUMAN GENECHIP, AFFYMETRIX), IMMUNOFLUORESCENT IN SITU LABELING COUPLED WITH VIDEO-MICROSCOPY RECORDING/ANALYSES, CHROMATIN-IMMUNOPRECIPITATION (CHIP), POLYMERASE CHAIN REACTIONS (PCR), REAL-TIME QUANTITATIVE PCR (RT-QPCR) AND BISULFITE GENOME SEQUENCING WERE THE MAIN METHODS APPLIED. STATISTICAL ANALYSES WERE PERFORMED BY APPLYING GCRMA AND SAM ANALYSIS (MICROARRAY DATA) AND STUDENT'S T-TEST OR MANN & WHITNEY'S U-TEST. RESULTS: HEREIN WE SHOW THAT, REMARKABLY, IN A RESISTANT MALE CLL CELLS THE VAST MAJORITY OF GENES WERE DOWN-REGULATED COMPARED WITH SENSITIVE CELLS, WHEREAS THIS WAS NOT THE CASE IN CELLS DERIVED FROM FEMALES. THIS GENE DOWN-REGULATION WAS FOUND TO BE ASSOCIATED WITH AN OVERALL GAIN OF HETEROCHROMATIN AS EVIDENCED BY IMMUNOFLUORESCENT LABELING OF HETEROCHROMATIN PROTEIN 1ALPHA (HP-1), TRIMETHYLATED HISTONE 3 LYSINE 9 (3METH3K9), AND 5-METHYLCYTIDINE (5METC). NOTABLY, 17 GENES WERE FOUND TO BE COMMONLY DEREGULATED IN RESISTANT MALE AND FEMALE CELL SAMPLES. AMONG THESE, RELB WAS IDENTIFIED AS A DISCRIMINATORY CANDIDATE GENE REPRESSED IN THE MALE AND UPREGULATED IN THE FEMALE RESISTANT CELLS. CONCLUSION: THE MOLECULAR DEFECTS IN THE SILENCING OF RELB INVOLVE AN INCREASE IN H3K9- BUT NOT CPG-ISLAND METHYLATION IN THE PROMOTER REGIONS. INCREASE IN ACETYL-H3 IN RESISTANT FEMALE BUT NOT MALE CLL SAMPLES AS WELL AS A DECREASE OF TOTAL CELLULAR LEVEL OF RELB AFTER AN INHIBITION OF HISTONE DEACETYLASE (HDAC) BY TRICHOSTATIN A (TSA), FURTHER EMPHASIZE THE ROLE OF EPIGENETIC MODIFICATIONS WHICH COULD DISCRIMINATE TWO CLL SUBSETS. TOGETHER, THESE RESULTS HIGHLIGHTED THE EPIGENETIC RELB SILENCING AS A NEW MARKER OF THE PROGRESSIVE DISEASE IN MALES.	2010	
                                                                                                                                                                                                                                                                                                                                                                                      
8 5273  29 PROMOTER METHYLATION AND BDNF AND DAT1 GENE EXPRESSION PROFILES IN PATIENTS WITH DRUG ADDICTION. BACKGROUND: DRUG ADDICTION IS A BRAIN DISORDER THAT HAS NEGATIVE CONSEQUENCES FOR INDIVIDUALS AND SOCIETY. ADDICTIONS ARE CHRONIC RELAPSING DISEASES OF THE BRAIN THAT ARE CAUSED BY DIRECT DRUG-INDUCED EFFECTS AND PERSEVERING NEUROADAPTATIONS AT THE EPIGENETIC, NEUROPEPTIDE AND NEUROTRANSMITTER LEVELS. BECAUSE THE DOPAMINERGIC SYSTEM HAS A SIGNIFICANT ROLE IN DRUG ABUSE, THE PURPOSE OF THIS STUDY WAS TO ANALYZE THE METHYLATION AND EXPRESSION PROFILE OF BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) AND DOPAMINE TRANSPORTER (DAT1) GENES IN INDIVIDUALS WITH DRUG ADDICTION. MATERIALS AND METHODS: BDNF AND DAT1 PROMOTER METHYLATION WERE INVESTIGATED WITH A METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (PCR) TECHNIQUE IN BLOOD SAMPLES FROM 75 INDIVIDUALS WITH DRUG ADDICTION AND 65 HEALTHY CONTROLS. THE EXPRESSION LEVELS OF BDNF AND DAT1 WERE ASSESSED IN 12 MRNA SAMPLES FROM THE BLOOD OF PATIENTS AND COMPARED TO THE SAMPLES OF HEALTHY CONTROLS (N = 12) WITH REAL-TIME QUANTITATIVE REVERSE TRANSCRIPTION PCR. RESULTS: NO SIGNIFICANT DIFFERENCES WERE FOUND IN THE METHYLATION OF BDNF AND DAT1 BETWEEN PATIENTS AND CONTROLS, BUT THE RELATIVE LEVELS OF EXPRESSION OF BDNF AND DAT1 MRNA DIFFERED SIGNIFICANTLY IN THE PATIENTS COMPARED TO CONTROLS (P < 0.0001). CONCLUSION: THESE RESULTS SHOWED THAT THE METHYLATION STATUS OF THE BDNF AND DAT1 GENES HAD NO SIGNIFICANT FUNCTION IN THE PROCESSES OF DRUG ADDICTION.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
9 3122  31 GESTATIONAL VALPROIC ACID EXPOSURE INDUCES EPIGENETIC MODIFICATIONS IN MURINE DECIDUA. INTRODUCTION: VALPROIC ACID (VPA), A WIDELY PRESCRIBED ANTIEPILEPTIC DRUG AND AN EFFECTIVE TREATMENT FOR BIPOLAR DISORDER AND NEUROPATHIC PAIN, RESULTS IN MULTIPLE DEVELOPMENTAL DEFECTS FOLLOWING IN UTERO EXPOSURE. UTERINE DECIDUA PROVIDES NUTRITIONAL AND PHYSICAL SUPPORT DURING IMPLANTATION AND EARLY EMBRYONIC DEVELOPMENT. PERTURBATIONS IN THE MOLECULAR MECHANISMS WITHIN DECIDUAL TISSUE DURING EARLY PREGNANCY MIGHT AFFECT EARLY EMBRYONIC GROWTH, RESULT IN EARLY PREGNANCY LOSS OR CAUSE COMPLICATIONS IN THE LATER GESTATIONAL STAGE. VPA IS A KNOWN HISTONE DEACETYLASE INHIBITOR AND EPIGENETIC CHANGES SUCH AS HISTONE HYPERACETYLATION AND METHYLATION HAVE BEEN PROPOSED AS A MECHANISM OF VPA-INDUCED TERATOGENESIS. METHODS: THIS STUDY INVESTIGATED THE EFFECTS OF IN UTERO VPA EXPOSURE ON HISTONE MODIFICATIONS IN MURINE DECIDUAL TISSUE. PREGNANT CD-1 MICE WERE EXPOSED TO 400 MG/KG VPA OR SALINE ON GD9 VIA SUBCUTANEOUS INJECTION. DECIDUAL TISSUE FROM EACH GESTATIONAL SAC WAS HARVESTED AT 1, 3 AND 6 H FOLLOWING EXPOSURE. LEVELS OF ACETYLATED HISTONES H3, H4 AND H3K56, AS WELL AS METHYLATED HISTONES H3K9 AND H3K27 WERE ACID EXTRACTED AND ASSESSED BY WESTERN BLOTTING FOLLOWED BY ACID HISTONE EXTRACTION. RESULTS: VPA EXPOSURE INDUCED A SIGNIFICANT INCREASE (P < 0.05) IN THE LEVELS OF ACETYLATED H3 AT 1, 3 H; ACETYLATED H4 AT 1, 3 AND 6 H AND TRIMETHYLATED H3K9 AT 6 H. IN CONTRAST, NO SIGNIFICANT PERTURBATIONS WERE NOTED IN THE LEVELS OF MONOMETHYLATED H3K9, TRIMETHYLATED H3K27 AND ACETYLATED H3K56. DISCUSSION: THE RESULTS FROM THIS STUDY SUGGEST THAT VPA-INDUCED DECIDUAL HISTONE MODIFICATIONS MIGHT PLAY AN IMPORTANT ROLE AS A MECHANISM OF VPA-INDUCED TERATOGENESIS DURING EARLY EMBRYONIC GROWTH.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
10 2030  33 EPIGENETIC CHANGES IN INFLAMMATORY GENES AND THE PROTECTIVE EFFECT OF COOKED RHUBARB ON PANCREATIC TISSUE OF RATS WITH CHRONIC ALCOHOL EXPOSURE. CHRONIC ALCOHOL CONSUMPTION, WHICH IS OBSERVED WORLDWIDE, CAN DAMAGE PANCREATIC TISSUE AND PROMOTE PANCREATITIS. RHUBARB IS A WIDELY USED TRADITIONAL CHINESE HERBAL MEDICINE FOR TREATING PANCREATITIS IN CHINA. HOWEVER, FEW PHARMACOLOGICAL STUDIES HAVE INVESTIGATED ITS EPIGENETIC REGULATION. IN THIS STUDY, WE INVESTIGATED WHETHER CHRONIC EXPOSURE TO ALCOHOL CAN ALTER INFLAMMATORY GENE EXPRESSION AND THE EPIGENETIC REGULATION EFFECT OF COOKED RHUBARB IN THE PANCREATIC TISSUE OF RATS. FIRST, CHANGES IN INFLAMMATORY CYTOKINE DNA METHYLATION (IL-10, IL-1ALPHA, TNF-ALPHA, NF-KAPPAB AND TGF-BETA) WERE DETECTED IN PANCREATIC TISSUE OF SPRAGUE-DAWLEY RATS WITH VARYING ALCOHOL EXPOSURE TIMES (4, 6, 8, OR 12 WEEKS), AND THEN WITH VARYING DOSES OF COOKED RHUBARB TREATMENT (3, 6, OR 12 G/DAY). DNA METHYLATION LEVELS, RELATED RNA CONCENTRATIONS AND PROTEIN EXPRESSION OF SPECIFIC INFLAMMATORY CYTOKINES, AND HISTOPATHOLOGICAL SCORE WERE ANALYSED IN PANCREATIC TISSUE OF SPRAGUE-DAWLEY RATS. THE RESULTS SHOWED THAT CHRONIC ALCOHOL EXPOSURE (8 WEEKS) REDUCED THE LEVEL OF IL-1ALPHA DNA METHYLATION AND INCREASED ITS PROTEIN EXPRESSION IN ACINAR CELLS (P < 0.05). IN THE ACINAR CELLS, THE LEVEL OF IL-10 DNA METHYLATION INCREASED, RESULTING IN A REDUCTION OF PROTEIN EXPRESSION (P < 0.05). SIMULTANEOUSLY, CHRONIC ALCOHOL EXPOSURE INCREASED THE PATHOLOGICAL DAMAGE TO THE PANCREAS (P < 0.05). FINALLY, COOKED RHUBARB TREATMENT (3 G/KG/DAY) EFFECTIVELY ALLEVIATED THESE CHANGES IN PANCREATIC TISSUE FROM CHRONIC ALCOHOL EXPOSURE (P < 0.05). THESE RESULTS INDICATE THAT CHRONIC EXPOSURE TO ALCOHOL LEADS TO CHANGES IN DNA METHYLATION AND PROTEIN EXPRESSION OF INFLAMMATORY GENES, AND COOKED RHUBARB MAY HAVE A PROTECTIVE EFFECT ON THE PANCREATIC TISSUE OF RATS.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
11 2590  38 EPIGENETICS OF PROTEASOME INHIBITION IN THE LIVER OF RATS FED ETHANOL CHRONICALLY. AIM: TO EXAMINE THE EFFECTS OF ETHANOL-INDUCED PROTEASOME INHIBITION, AND THE EFFECTS OF PROTEASOME INHIBITION IN THE REGULATION OF EPIGENETIC MECHANISMS. METHODS: RATS WERE FED ETHANOL FOR 1 MO USING THE TSUKAMOTO-FRENCH MODEL AND WERE COMPARED TO RATS GIVEN THE PROTEASOME INHIBITOR PS-341 (BORTEZOMIB, VELCADE(TM)) BY INTRAPERITONEAL INJECTION. MICROARRAY ANALYSIS AND REAL TIME PCR WERE PERFORMED AND PROTEASOME ACTIVITY ASSAYS AND WESTERN BLOT ANALYSIS WERE PERFORMED USING ISOLATED NUCLEI. RESULTS: CHRONIC ETHANOL FEEDING CAUSED A SIGNIFICANT INHIBITION OF THE UBIQUITIN PROTEASOME PATHWAY IN THE NUCLEUS, WHICH LED TO CHANGES IN THE TURNOVER OF TRANSCRIPTIONAL FACTORS, HISTONE-MODIFYING ENZYMES, AND, THEREFORE, AFFECTED EPIGENETIC MECHANISMS. CHRONIC ETHANOL FEEDING WAS RELATED TO AN INCREASE IN HISTONE ACETYLATION, AND IT IS HYPOTHESIZED THAT THE PROTEASOME PROTEOLYTIC ACTIVITY REGULATED HISTONE MODIFICATIONS BY CONTROLLING THE STABILITY OF HISTONE MODIFYING ENZYMES, AND, THEREFORE, REGULATED THE CHROMATIN STRUCTURE, ALLOWING EASY ACCESS TO CHROMATIN BY RNA POLYMERASE, AND, THUS, PROPER GENE EXPRESSION. PROTEASOME INHIBITION BY PS-341 INCREASED HISTONE ACETYLATION SIMILAR TO CHRONIC ETHANOL FEEDING. IN ADDITION, PROTEASOME INHIBITION CAUSED DRAMATIC CHANGES IN HEPATIC REMETHYLATION REACTIONS AS THERE WAS A SIGNIFICANT DECREASE IN THE ENZYMES RESPONSIBLE FOR THE REGENERATION OF S-ADENOSYLMETHIONINE, AND, IN PARTICULAR, A SIGNIFICANT DECREASE IN THE BETAINE-HOMOCYSTEINE METHYLTRANSFERASE ENZYME. THIS SUGGESTED THAT HYPOMETHYLATION WAS ASSOCIATED WITH PROTEASOME INHIBITION, AS INDICATED BY THE DECREASE IN HISTONE METHYLATION. CONCLUSION: THE ROLE OF PROTEASOME INHIBITION IN REGULATING EPIGENETIC MECHANISMS, AND ITS LINK TO LIVER INJURY IN ALCOHOLIC LIVER DISEASE, IS THUS A PROMISING APPROACH TO STUDY LIVER INJURY DUE TO CHRONIC ETHANOL CONSUMPTION.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
12 1504  29 DNA METHYLATION AND HISTONE DEACETYLATION REGULATING INSULIN SENSITIVITY DUE TO CHRONIC COLD EXPOSURE. IN THIS STUDY, WE INVESTIGATED THE CAUSAL RELATIONSHIP BETWEEN CHRONIC COLD EXPOSURE AND INSULIN RESISTANCE AND THE MECHANISMS OF HOW DNA METHYLATION AND HISTONE DEACETYLATION REGULATE COLD-REDUCED INSULIN RESISTANCE. 46 ADULT MALE MICE FROM POSTNATAL DAY 90-180 WERE RANDOMLY ASSIGNED TO CONTROL GROUP AND COLD-EXPOSURE GROUP. MICE IN COLD-EXPOSURE GROUP WERE PLACED AT TEMPERATURE FROM -1 TO 4 DEGREES C FOR 30 DAYS TO MIMIC CHRONIC COLD ENVIRONMENT. THEN, FASTING BLOOD GLUCOSE, BLOOD INSULIN LEVEL AND INSULIN RESISTANCE INDEX WERE MEASURED WITH ENZYMATIC METHODS. IMMUNOFLUORESCENT LABELING WAS CARRIED OUT TO VISUALIZE THE INSULIN RECEPTOR SUBSTRATE 2 (IRS2), OBESE RECEPTOR (OB-R, A LEPTIN RECEPTOR), VOLTAGE-DEPENDENT ANION CHANNEL PROTEIN 1 (VDAC1), CYTOCHROME C (CYTC), 5-METHYLCYTOSINE (5-MC) POSITIVE CELLS IN HIPPOCAMPAL CA1 AREA. FURTHERMORE, THE EXPRESSIONS OF SOME PROTEINS MENTIONED ABOVE WERE DETECTED WITH WESTERN BLOT. THE RESULTS SHOWED: 1 IN CIRCLE CHRONIC COLD EXPOSURE COULD REDUCE THE INSULIN RESISTANCE INDEX (P < 0.01) AND INCREASE THE NUMBER OF IRS2 POSITIVE CELLS AND OB-R POSITIVE CELLS IN HIPPOCAMPUS (P < 0.01). 2 IN CIRCLE THE EXPRESSIONS OF MITOCHONDRIAL ENERGY-RELATIVE PROTEINS, VDAC1 AND CYTC, WERE HIGHER IN COLD-EXPOSURE GROUP THAN IN CONTROL GROUP WITH BOTH IMMUNOHISTOCHEMICAL STAINING AND WESTERN BLOT (P < 0.01). 3 IN CIRCLE CHRONIC COLD EXPOSURE INCREASED DNA METHYLATION AND HISTONE DEACETYLATION IN THE PYRAMIDAL CELLS OF CA1 AREA AND LED TO AN INCREASE IN THE EXPRESSION OF HISTONE DEACETYLASE 1 (HDAC1) AND DNA METHYLATION RELATIVE ENZYMES (P < 0.01). IN CONCLUSION, CHRONIC COLD EXPOSURE CAN IMPROVE INSULIN SENSITIVITY, WITH THE INVOLVEMENT OF DNA METHYLATION, HISTONE DEACETYLATION AND THE REGULATION OF MITOCHONDRIAL ENERGY METABOLISM. THESE EPIGENETIC MODIFICATIONS PROBABLY FORM THE BASIC MECHANISM OF COLD-REDUCED INSULIN RESISTANCE.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
13 1328  27 DEPRESSION AND STRESS LEVELS INCREASE RISK OF LIVER CANCER THROUGH EPIGENETIC DOWNREGULATION OF HYPOCRETIN. RECENT STUDIES SUGGEST THAT HYPOCRETIN (HCRT, OREXIN) ARE INVOLVED IN STRESS REGULATION OF DEPRESSION THROUGH THE HYPOTHALAMIC-PITUITARY-ADRENAL (HPA) AXIS. HOWEVER, THE MOLECULAR MECHANISM BY WHICH HYPOCRETIN REGULATE NEUROBIOLOGICAL RESPONSES IS UNKNOWN. HEREIN, THE EFFECTS OF CHRONIC STRESS ON THE EPIGENETIC MODIFICATION OF HCRT AND ITS ASSOCIATION WITH DEPRESSION WERE EXPLORED WITH REGARD TO A POTENTIAL ROLE IN CANCER PROGRESSION. IN THE STUDY, SPRAGUE DAWLEY (SD) RATS WERE USED TO ESTABLISH AN ANIMAL MODEL OF CANCER WITH DEPRESSION BY ADMINISTRATING N-NITROSODIETHYLAMINE (DEN) AND CHRONIC UNPREDICTABLE MILD STRESS (CUMS). RNA-SEQUENCING WAS USED TO DETECT DIFFERENTIALLY EXPRESSED GENES IN THE HIPPOCAMPUS OF RATS AND QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QRT-PCR) WAS USED TO VALIDATE THE RESULTS OF RNA-SEQUENCING. THE STATUS OF HCRT PROMOTER METHYLATION WAS ASSESSED BY METHYLATION SPECIFIC POLYMERASE CHAIN REACTION. BEHAVIORAL TESTS SHOWED THAT RATS EXPOSED TO CUMS HAD SIGNIFICANT DEPRESSIVE-LIKE BEHAVIORS. THE NUMBER OF LIVER TUMORS AND TUMOR LOAD IN DEPRESSED RATS EXPOSED TO CUMS WAS HIGHER THAN IN SD RATS WITHOUT CUMS. RNA-SEQUENCING REVEALED THAT HCRT WAS ONE OF THE MOST SIGINIFICANTLY DOWNREGULATED GENE IN THE HIPPOCAMPUS OF SD RATS WITH CUMS COMPARED TO NON-STRESSED GROUP, WHICH WAS VALIDATED BY QRT-PCR. HCRT MRNA EXPRESSION WAS DOWNREGULATED AND THE PROMOTER FOR HCRT WAS HYPER-METHYLATED IN THOSE WITH DEPRESSION. THESE RESULTS IDENTIFIED A CRITICAL ROLE FOR CHRONIC PSYCHOLOGICAL STRESSORS IN TUMORIGENESIS AND CANCER PROGRESSION, VIA EPIGENETIC HCRT DOWNREGULATION. SUCH EPIGENETIC DOWNREGULATION MAY BE THE MOLECULAR BASIS FOR THE ASSOCIATION OF CANCER WITH DEPRESSION.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
14 6148  39 THE EXPRESSION OF TRANSCRIPTION FACTORS MECP2 AND CREB IS MODULATED IN INFLAMMATORY PELVIC PAIN. EARLY ACTIVATION OF TRANSCRIPTION FACTORS IS ONE OF THE EPIGENETIC MECHANISMS CONTRIBUTING TO THE INDUCTION AND MAINTENANCE OF CHRONIC PAIN STATES. PREVIOUS STUDIES IDENTIFIED THE CHANGES IN A NUMBER OF NOCICEPTION-RELATED GENES, SUCH AS CALCITONIN GENE-RELATED PEPTIDE (CGRP), SUBSTANCE P (SP), AND BRAIN-DERIVED NEUROTROPIC FACTOR (BDNF) IN THE PELVIC ORGANS AFTER TRANSIENT COLONIC INFLAMMATION. THE GENE AND PROTEIN EXPRESSION OF THESE NEUROPEPTIDES COULD BE MODULATED BY TRANSCRIPTION FACTORS METHYL-CPG-BINDING PROTEIN 2 (MECP2) AND CAMP RESPONSE ELEMENT-BINDING PROTEIN (CREB). IN THIS STUDY, WE AIMED TO EVALUATE TIME-DEPENDENT CHANGES IN THE EXPRESSION LEVELS OF MECP2 AND CREB IN THE LUMBOSACRAL (LS) SPINAL CORD AND SENSORY GANGLIA AFTER INFLAMMATION-INDUCED PELVIC PAIN IN RAT. ADULT SPRAGUE-DAWLEY RATS WERE TREATED WITH 2,4,6-TRINITROBENZENESULFONIC ACID (TNBS) TO INDUCE TRANSIENT COLONIC INFLAMMATION. LS (L6-S2) SPINAL CORD SEGMENTS AND RESPECTIVE DORSAL ROOT GANGLIAS (DRGS) WERE ISOLATED FROM CONTROL AND EXPERIMENTAL ANIMALS AT 1, 2, 6, 24 H AND 3 DAYS POST-TNBS TREATMENT. IMMUNOHISTOCHEMICAL (IHC) LABELING AND WESTERN BLOTTING EXPERIMENTS WERE PERFORMED TO ASSESS THE EXPRESSION OF MECP2, CREB AND THEIR PHOSPHORYLATED FORMS. TOTAL MECP2 EXPRESSION, BUT NOT PHOSPHORYLATED P-MECP2 (PS421MECP2) EXPRESSION WAS DETECTED IN THE CELLS OF THE SPINAL DORSAL HORN UNDER CONTROL CONDITIONS. COLONIC INFLAMMATION TRIGGERED A SIGNIFICANT DECREASE IN THE NUMBER OF MECP2-EXPRESSING NEURONS IN PARALLEL WITH ELEVATED NUMBERS OF PS421MECP2-EXPRESSING CELLS AT 2 H AND 6 H POST-TNBS. THE MAJORITY OF MECP2-POSITIVE CELLS (80 +/- 6%) CO-EXPRESSED CREB. TNBS TREATMENT CAUSED A TRANSIENT UP-REGULATION OF CREB-EXPRESSING CELLS AT 1 H POST-TNBS ONLY. THE NUMBER OF CELLS EXPRESSING PHOSPHORYLATED CREB (PS133CREB) DID NOT CHANGE AT 1 H AND 2 H POST-TNBS, BUT WAS DOWN-REGULATED BY THREE FOLDS AT 6 H POST-TNBS. ANALYSIS OF DRG SECTIONS REVEALED THAT THE NUMBER OF MECP2-POSITIVE NEURONS WAS UP-REGULATED BY TNBS TREATMENT, REACHING THREE-FOLD INCREASE AT 2 H POST-TNBS, AND EIGHT-FOLD INCREASE AT 6 H POST-TNBS (P </= 0.05 TO CONTROL). THESE DATA SHOWED EARLY CHANGES IN MECP2 AND CREB EXPRESSION IN THE DORSAL HORN OF THE SPINAL CORD AND SENSORY GANGLIA AFTER COLONIC INFLAMMATION, SUGGESTING A POSSIBLE CONTRIBUTION MECP2 AND CREB SIGNALING IN THE DEVELOPMENT OF VISCERAL HYPERALGESIA AND PELVIC PAIN FOLLOWING PERIPHERAL INFLAMMATION.	2018	

15 3792  26 INTERLEUKIN-1BETA INCREASES THE RISK OF GASTRIC CANCER THROUGH INDUCTION OF ABERRANT DNA METHYLATION IN A MOUSE MODEL. INTERLEUKIN-1BETA (IL-1BETA) HAS A SIGNIFICANT ROLE IN CHRONIC GASTRIC INFLAMMATION AND MANIFESTATIONS OF GASTRIC DISEASES. THE PRESENT STUDY AIMED TO ELUCIDATE THE SPECIFIC ROLE OF IL-1BETA IN INDUCTION OF DNA METHYLATION USING IL-1 RECEPTOR TYPE 1 KNOCKOUT (IL-1R1(-)/(-)) MICE. IN THE PRESENT STUDY, WILD-TYPE (WT) AND IL-1R1(-)/(-) MICE WERE INJECTED WITH IL-1BETA (5 MICROG/KG/DAY). SERUM LEVELS OF IL-1BETA, INTERLEUKIN-6 (IL-6) AND NITRIC OXIDE (NO) WERE MEASURED BY ENZYME-LINKED IMMUNOSORBENT OR NO ASSAYS. E-CADHERIN (E-CAD) METHYLATION STATUS AND MESSENGER (M)RNA EXPRESSION OF IL-1BETA, IL-6, E-CAD AND INDUCIBLE NITRIC OXIDE SYNTHASE (INOS) WERE ANALYZED. RESULTS FROM THE PRESENT STUDY INDICATED SIGNIFICANTLY HIGHER IL-1BETA MRNA EXPRESSION (P<0.001) IN WT MICE COMPARED WITH IL-1R1(-)/(-) MICE. IL-1BETA AND IL-6 RELEASE WAS SIGNIFICANTLY INCREASED IN TREATED WT MICE COMPARED WITH IL-1R1(-)/(-) MICE AT 1 H, 4 H AND 8 H (ALL P<0.005). IL-1BETA RELEASE WAS ONLY DETECTED IN WT MICE FOLLOWING A SECOND DOSE MEASURED AT DAY 3, WEEK 1 AND WEEK 2 WHEN COMPARED WITH IL-1R1(-)/(-) MICE. PROMOTER METHYLATION OF E-CAD AND A DECREASE IN GENE EXPRESSION WAS OBSERVED IN TREATED WT MICE. MRNA EXPRESSION OF INOS IN WT MICE WAS SIGNIFICANTLY INCREASED AT WEEK 1 COMPARED WITH IL-1R1(-)/(-) MICE (P=0.0411). FURTHERMORE, A SIGNIFICANTLY INCREASED LEVEL OF NO PRODUCTION WAS OBSERVED IN TREATED WT MICE (P<0.005 AT 8 H AND WEEK 1; P<0.001 AT 4 H AND DAY 3) WHEN COMPARED WITH IL-1R1(-)/(-) MICE. THE PRESENT RESULTS INDICATED THAT IL-1BETA WAS ABLE TO DIRECTLY INDUCE DNA METHYLATION, WHICH MAY LINK INFLAMMATION-INDUCED EPIGENETIC CHANGES AND THE DEVELOPMENT OF GASTRIC DISEASES.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
16 2395  31 EPIGENETIC REPROGRAMMING IN MIST1(-/-) MICE PREDICTS THE MOLECULAR RESPONSE TO CERULEIN-INDUCED PANCREATITIS. GENE EXPRESSION IS AFFECTED BY MODIFICATIONS TO HISTONE CORE PROTEINS WITHIN CHROMATIN. CHANGES IN THESE MODIFICATIONS, OR EPIGENETIC REPROGRAMMING, CAN DICTATE CELL FATE AND PROMOTE SUSCEPTIBILITY TO DISEASE. THE GOAL OF THIS STUDY WAS TO DETERMINE THE EXTENT OF EPIGENETIC REPROGRAMMING IN RESPONSE TO CHRONIC STRESS THAT OCCURS FOLLOWING ABLATION OF MIST1 (MIST1(-/-) ), WHICH IS REPRESSED IN PANCREATIC DISEASE. CHROMATIN IMMUNOPRECIPITATION FOR TRIMETHYLATION OF LYSINE RESIDUE 4 ON HISTONE 3 (H3K4ME3) IN PURIFIED ACINAR CELLS FROM WILD TYPE AND MIST1(-/-) MICE WAS FOLLOWED BY NEXT GENERATION SEQUENCING (CHIP-SEQ) OR CHIP-QPCR. H3K4ME3-ENRICHED GENES WERE ASSESSED FOR EXPRESSION BY QRT-PCR IN PANCREATIC TISSUE BEFORE AND AFTER INDUCTION OF CERULEIN-INDUCED PANCREATITIS. WHILE MOST OF H3K4ME3-ENRICHMENT IS RESTRICTED TO TRANSCRIPTIONAL START SITES, >25% OF ENRICHMENT SITES ARE FOUND WITHIN, DOWNSTREAM OR BETWEEN ANNOTATED GENES. LESS THAN 10% OF THESE SITES WERE ALTERED IN MIST1(-/-) ACINI, WITH MOST CHANGES IN H3K4ME3 ENRICHMENT NOT REFLECTING ALTERED GENE EXPRESSION. INGENUITY PATHWAY ANALYSIS OF GENES DIFFERENTIALLY-ENRICHED FOR H3K4ME3 REVEALED AN ASSOCIATION WITH PANCREATITIS AND PANCREATIC DUCTAL ADENOCARCINOMA IN MIST1(-/-) TISSUE. MOST OF THESE GENES WERE NOT DIFFERENTIALLY EXPRESSED BUT SEVERAL WERE READILY INDUCED BY ACUTE EXPERIMENTAL PANCREATITIS, WITH SIGNIFICANTLY INCREASED EXPRESSION IN MIST1(-/-) TISSUE RELATIVE TO WILD TYPE MICE. WE SUGGEST THAT THE CHRONIC CELL STRESS OBSERVED IN THE ABSENCE OF MIST1 RESULTS IN EPIGENETIC REPROGRAMMING OF GENES INVOLVED IN PROMOTING PANCREATITIS TO A POISED STATE, THEREBY INCREASING THE SENSITIVITY TO EVENTS THAT PROMOTE DISEASE.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
17 4742  26 NOVEL HISTONE MODIFICATIONS IN MICROGLIA DERIVED FROM A MOUSE MODEL OF CHRONIC PAIN. AS THE RESIDENT IMMUNE CELLS IN THE CENTRAL NERVOUS SYSTEM, MICROGLIA PLAY AN IMPORTANT ROLE IN THE MAINTENANCE OF ITS HOMEOSTASIS. DYSREGULATION OF MICROGLIA HAS BEEN ASSOCIATED WITH THE DEVELOPMENT AND MAINTENANCE OF CHRONIC PAIN. HOWEVER, THE RELEVANT MOLECULAR PATHWAYS REMAIN POORLY DEFINED. IN THIS STUDY, WE USED A MASS SPECTROMETRY-BASED PROTEOMIC APPROACH TO SCREEN POTENTIAL CHANGES OF HISTONE PROTEIN MODIFICATIONS IN MICROGLIA ISOLATED FROM THE BRAIN OF CONTROL AND CISPLATIN-INDUCED NEUROPATHIC PAIN ADULT C57BL/6J MALE MICE. WE IDENTIFIED SEVERAL NOVEL MICROGLIAL HISTONE MODIFICATIONS ASSOCIATED WITH PAIN, INCLUDING STATISTICALLY SIGNIFICANTLY DECREASED HISTONE H3.1 LYSINE 27 MONO-METHYLATION (H3.1K27ME1, 54.8% OF CONTROL) AND H3 LYSINE 56 TRI-METHYLATION (7.5% OF CONTROL), AS WELL AS A TREND SUGGESTING INCREASED H3 TYROSINE 41 NITRATION. WE FURTHER INVESTIGATED THE FUNCTIONAL ROLE OF H3.1K27ME1 AND FOUND THAT TREATMENT OF CULTURED MICROGLIAL CELLS FOR 4 CONSECUTIVE DAYS WITH 1-10 MUM OF NCDM-64, A POTENT AND SELECTIVE INHIBITOR OF LYSINE DEMETHYLASE 7A, AN ENZYME RESPONSIBLE FOR THE DEMETHYLATION OF H3K27ME1, DOSE-DEPENDENTLY ELEVATED ITS LEVELS WITH A GREATER THAN A TWO-FOLD INCREASE OBSERVED AT 10 MUM COMPARED TO VEHICLE-TREATED CONTROL CELLS. MOREOVER, PRETREATMENT OF MICE WITH NCDM-64 (10 OR 25 MG/KG/DAY, I.P.) PRIOR TO CISPLATIN TREATMENT PREVENTED THE DEVELOPMENT OF NEUROPATHIC PAIN IN MICE. THE IDENTIFICATION OF SPECIFIC CHROMATIN MARKS IN MICROGLIA ASSOCIATED WITH CHRONIC PAIN MAY YIELD CRITICAL INSIGHT INTO THE CONTRIBUTION OF MICROGLIA TO THE DEVELOPMENT AND MAINTENANCE OF PAIN, AND OPENS NEW AVENUES FOR THE DEVELOPMENT OF NOVEL NONOPIOID THERAPEUTICS FOR THE EFFECTIVE MANAGEMENT OF CHRONIC PAIN.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
18 2297  31 EPIGENETIC REGULATION OF ACUTE INFLAMMATORY PAIN. ACUTE PAIN IS ASSOCIATED WITH TISSUE DAMAGE, WHICH RESULTS IN THE RELEASE OF INFLAMMATORY MEDIATORS. RECENT STUDIES POINT TO THE INVOLVEMENT OF EPIGENETIC MECHANISMS (DNA METHYLATION) IN THE DEVELOPMENT OF PAIN. WE HAVE FOUND THAT DURING ACUTE INFLAMMATORY PAIN INDUCED BY THE APPLICATION OF 10% MUSTARD OIL ON THE TONGUES OF RATS, LEVELS OF DNMT3A AND 3B WERE ELEVATED MARKEDLY (36 AND 42 % RESPECTIVELY), WHEREAS THE LEVEL OF DNMT1 WAS NOT CHANGED SIGNIFICANTLY. PREVIOUS INJECTION OF XEFOCAM WITH 0,4 MG/KG DOSE DECREASED LEVELS OF DNMT3A AND 3B (25 AND 24% RESPECTIVELY). THE LEVEL OF DNMT1 WAS NOT CHANGED SIGNIFICANTLY COMPARED TO THE CONTROL GROUP. THE FINDINGS SUPPORT THE IDEA THAT INHIBITORS OF DNA-METHYLTRANSFERASES COULD BE USEFUL FOR PAIN MANAGEMENT. OUR DATA SUGGEST THAT NSAIDS (ALONE OR IN COMBINATION WITH DNMT INHIBITORS) MAY BE PROPOSED AS POSSIBLE EPIGENETIC REGULATORY AGENTS, WHICH MAY PLAY A ROLE IN EPIGENETIC MECHANISMS INDIRECTLY THROUGH ALTERING THE ACTIVITY OF INFLAMMATORY MEDIATORS INVOLVED IN PAIN DEVELOPMENT.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
19 3986  38 LONG-TERM OVARIECTOMY INCREASES BDNF GENE METHYLATION STATUS IN MOUSE HIPPOCAMPUS. ESTRADIOL (E) HAS BEEN SUGGESTED TO HAVE A NEUROPROTECTIVE EFFECT IN YOUNG ANIMALS BUT HAS NEUTRAL OR HARMFUL EFFECTS WHEN IT IS ADMINISTERED TO AGED ANIMALS. IN THE PRESENT STUDY, WE DETERMINED WHETHER THE POST-OVARIECTOMY (POST-OVX) TIMEFRAME ELAPSED BEFORE THE INITIATION OF CHRONIC E TREATMENT IS CRITICAL FOR THE ESTROGENIC INDUCTION OF NEUROTROPHINS (BRAIN-DERIVED NEUROTROPHIC FACTOR, BDNF, AND SYNAPTOPHYSIN, SYN) IN THE RODENT HIPPOCAMPUS. ADULT MICE WERE OVX AND, A SHORT PERIOD (SHORT-TERM E (STE) ANIMALS) OR A LONG PERIOD (LONG-TERM E (LTE) ANIMALS) AFTER THE OVX, WERE DAILY TREATED WITH E. CONTROL ANIMALS WERE TREATED WITH SESAME OIL (SHORT-TERM CONTROL (STC) AND LONG-TERM CONTROL (LTC) ANIMALS). PROTEIN EXPRESSION WAS DETERMINED USING AN IMMUNOHISTOCHEMICAL APPROACH. TRANSCRIPTIONAL ACTIVITY IN THE HIPPOCAMPUS OF INDIVIDUAL BDNF PROMOTERS WAS ASSESSED BY REAL-TIME QUANTITATIVE RT-PCR, AND THE METHYLATION LEVELS OF REGULATORY REGIONS WERE ANALYZED BY METHYLATION-SPECIFIC PCR AND COMBINED BISULFITE RESTRICTION ANALYSIS. STE ANIMALS SHOWED INCREASED BDNF AND SYN PROTEIN EXPRESSION AND A HIGHER ACTIVITY OF BDNF II, IV, AND V PROMOTERS. IN CONTRAST, LTE ANIMALS DID NOT SHOW E INDUCTION OF NEUROTROPHINS. IN THESE ANIMALS, THE METHYLATION LEVELS OF REGULATORY SEQUENCES OF THE BDNF WERE HIGHER THAN IN THE STE ANIMALS IN A CPG ISLAND OF PROMOTER V AND IN THE CRE REGULATORY SITE LOCATED IN PROMOTER IV. WITH THIS EXPERIMENT, WE DETERMINED THAT A PROLONGED PERIOD OF HYPOESTROGENICITY DISRUPTS THE E-INDUCTION OF NEUROTROPHINS, AND WE POSTULATED THAT DNA METHYLATION IS ONE OF THE EPIGENETIC MECHANISMS THAT COULD EXPLAIN THE E-INSENSITIVITY OF THE BDNF AFTER A LONG PERIOD POST-OVX.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
20 2714  32 EXERCISE-CONDITIONED PLASMA ATTENUATES NUCLEAR CONCENTRATIONS OF DNA METHYLTRANSFERASE 3B IN HUMAN PERIPHERAL BLOOD MONONUCLEAR CELLS. DNA METHYLATION IS MODIFIABLE BY ACUTE AND CHRONIC EXERCISE. DNA METHYLTRANSFERASES (DNMT) CATALYZE THIS PROCESS; HOWEVER, THERE IS A LACK OF LITERATURE CONCERNING THE SPECIFIC MECHANISMS BY WHICH EXERCISE-INDUCED MODIFICATIONS OCCUR. INTERLEUKIN 6 (IL-6) STIMULATION OF VARIOUS CELL LINES HAS BEEN SHOWN TO AUGMENT DNMT EXPRESSION AND NUCLEAR TRANSLOCATION, WHICH SUGGESTS A POSSIBLE PATHWAY BY WHICH EXERCISE IS ABLE TO ELICIT CHANGES IN EPIGENETIC ENZYMES. THE PRESENT STUDY SOUGHT TO ELUCIDATE THE RESPONSE OF THE DE NOVO METHYLTRANSFERASES DNMT3A AND DNMT3B TO CIRCULATORY FACTORS FOUND IN PLASMA ISOLATED FROM WHOLE BLOOD BEFORE AND AFTER 120-MIN OF TREADMILL RUNNING AT AN INTENSITY OF 60% OF INDIVIDUAL VELOCITY AT V O2MAX (VV O2MAX) INTERSPERSED WITH 30-SEC SPRINTS AT 90% OF VV O2MAX EVERY 10-MIN. PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) ISOLATED FROM A RESTING PARTICIPANT WERE INCUBATED WITH PLASMA ISOLATED FROM EXERCISING PARTICIPANTS (N = 10) OR RECOMBINANT IL-6 (RIL-6), FOLLOWED BY NUCLEAR PROTEIN EXTRACTION AND QUANTIFICATION OF DNMT3A AND DNMT3B CONCENTRATIONS. NUCLEAR CONCENTRATIONS OF DNMT3B SIGNIFICANTLY DECREASED FOLLOWING THE EXPERIMENTAL PROTOCOL (P = 0.03), WITH NO CHANGE OBSERVED IN DNMT3A (P = 0.514).VARIOUS CONCENTRATIONS OF RIL-6 CAUSED AN ELEVATION IN BOTH DNMT3A AND DNMT3B NUCLEAR CONCENTRATION COMPARED WITH THE BLANK CONTROL. THE CONFLICTING RESULTS BETWEEN EXERCISING AND RIL-6 CONDITIONS SUGGESTS THAT IL-6 DOES REGULATE DNMT NUCLEAR TRANSPORT, HOWEVER, OTHER PLASMA MEDIATORS MAY ALSO EXERT SIGNIFICANT INFLUENCE ON THE NUCLEAR CONCENTRATIONS OF THESE ENZYMES.	2015