1 5348 126 RASGRF1, A POTENTIAL METHYLATIC MEDIATOR OF ANTI-EPILEPTOGENESIS? EPILEPTOGENESIS, INDUCED BY STATUS EPILEPTICUS (SE), IS A CHRONIC PROCESS, AND INTERVENTION IN THIS PROGRESS MAY PREVENT CHRONIC EPILEPSY. IT HAS BEEN PROPOSED THAT DNA METHYLATION MIGHT BE RELATED WITH EPILEPTOGENESIS. RASGRF1 HAS A DIFFERENTIALLY METHYLATED REGION AT THE PROMOTER WHICH CAN SILENCE GENE EXPRESSION. WE HAVE PREVIOUSLY OBSERVED THE DOWN-REGULATION OF RASGRF1 IN EPILEPSY PATIENTS AND PROVED THAT HYPERMETHYLATION OF RASGRF1 REACHES MAXIMAL LEVEL AT THE LATENT PERIOD IN MICE AFTER KAINATE-INDUCED SE (KA MICE), WITH CORRESPONDING ALTERATION OF RASGRF1 EXPRESSION. IN THE PRESENT STUDY, N-PHTHALYL-L-TRYPTOPHAN (RG108), A DNA METHYLTRANSFERASE INHIBITOR, WAS APPLIED IN KA MICE AT LATENT PHASE AND THE BEHAVIOR, ELECTROENCEPHALOGRAM AND PATHOLOGICAL CHANGES WERE OBSERVED IN CHRONIC PHASE. METHYLATION AND EXPRESSION OF RASGRF1 WERE DETERMINED BY POLYMERASE CHAIN REACTION (PCR), WESTERN BLOTTING, AND BISULFITE SEQUENCING PCR. THE RESULTS SHOWED THAT THE INCIDENCE OF SPONTANEOUS RECURRENT SEIZURES (SRS) WAS SIGNIFICANTLY LOWER IN THE RG108 GROUP THAN THE NORMAL SALINE (NS) GROUP. SUBGROUP ANALYSIS SHOWED SIGNIFICANT HYPERMETHYLATION AND LOWER EXPRESSION OF RASGRF1 IN THE RG108-SRS SUBGROUP AND THE NS-SRS SUBGROUP BUT NOT IN THE RG108-NSRS (NO SRS) SUBGROUP AND THE NS-NSRS SUBGROUP COMPARED WITH THE CONTROL GROUP. NO SIGNIFICANT DIFFERENCE WAS FOUND BETWEEN THE RG108-SRS AND NS-SRS SUBGROUPS. MEANWHILE, HIPPOCAMPAL NEURONAL LOSS WAS OBSERVED IN RG108-SRS AND NS-SRS SUBGROUPS. WE THUS DEMONSTRATED THAT RG108 COULD MODIFY THE PROGRESSION OF EPILEPTOGENESIS AFTER KA INDUCED SE AND PREVENT CHRONIC EPILEPSY. MEANWHILE, HYPERMETHYLATION OF RASGRF1 AFTER KA INDUCED SE COULD BE REVERSED WITH CORRESPONDING CHANGES OF RASGRF1 EXPRESSION. ADDITIONALLY, WE SPECULATED THAT RASGRF1 MIGHT BE A POTENTIAL EPIGENETIC MEDIATOR IN EPILEPTOGENESIS AND CHRONIC EPILEPSY.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
2  510  64 ASSOCIATION OF RASGRF1 METHYLATION WITH EPILEPTIC SEIZURES. DNA METHYLATION, ONE OF THE MECHANISMS OF EPIGENETIC REGULATION, HAS BEEN SUGGESTED TO BE RELATED WITH EPILEPSY. RASGRF1 IS A PATERNALLY IMPRINTED GENE AND HAS A DIFFERENTIALLY METHYLATED REGION (DMR) AT THE PROMOTER THAT CAN SILENCE GENE EXPRESSION. WE HAVE PREVIOUSLY OBSERVED THE DOWN-REGULATION OF RASGRF1 IN THE TEMPORAL NEOCORTEX OF EPILEPSY PATIENTS AND IN THE HIPPOCAMPUS OF EPILEPTIC ANIMALS. HERE, WE FURTHER EXPLORED THE DYNAMIC CHANGE (1-DAY ACUTE PERIOD, 10-DAY LATENT PERIOD AND 45-DAY CHRONIC PHASE) OF DNA METHYLATION AND RASGRF1 EXPRESSION AFTER ACUTE EPILEPTIC SEIZURES IN KAINIC ACID (KA)-TREATED MICE, AND WE OBSERVED THE IMPACT OF N-PHTHALYL-L-TRYPTOPHAN (RG108), A DNA METHYLTRANSFERASE (DNMT) INHIBITOR, ON AN ACUTE EPILEPTIC MODEL BY POLYMERASE CHAIN REACTION (PCR), WESTERN BLOTTING, AND BISULFITE SEQUENCING PCR (BSP). THE RESULTS DIRECTLY SHOWED THAT THE METHYLATION OF THE RASGRF1 PROMOTER GRADUALLY INCREASED AND REACHED A MAXIMAL LEVEL AT THE LATENT PERIOD, WITH SUBSEQUENT SUPPRESSION OF RASGRF1 MRNA AND PROTEIN EXPRESSION LEVELS, WHICH REACHED A MINIMUM LEVEL IN THE CHRONIC PHASE. RG108 INHIBITED THE INCREASED METHYLATION OF THE RASGRF1 GENE, WITH SIGNIFICANT INHIBITION OCCURRING AT THE LATENT PERIOD, AND RESTORED RASGRF1 EXPRESSION LEVELS IN THE CHRONIC PHASE. IN ADDITION, WE DEMONSTRATED THAT RG108 COULD SUPPRESS ACUTE EPILEPTIC SEIZURES IN KA-TREATED MICE AND EPILEPTIC DISCHARGES IN 4-AMINOPYRIDINE (4-AP)-TREATED HIPPOCAMPAL SLICES. THESE FINDINGS DEMONSTRATE THAT RASGRF1 IS CLOSELY ASSOCIATED WITH EPILEPSY VIA THE ABERRANT METHYLATION OF RASGRF1, AND REGULATING THE METHYLATION STATUS OF RELEVANT GENES MIGHT BE AN INTRIGUING TOPIC IN FUTURE RESEARCH ON EPILEPSY.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
3 1425  36 DIFFERENTIAL DNA METHYLATION PROFILES OF CODING AND NON-CODING GENES DEFINE HIPPOCAMPAL SCLEROSIS IN HUMAN TEMPORAL LOBE EPILEPSY. TEMPORAL LOBE EPILEPSY IS ASSOCIATED WITH LARGE-SCALE, WIDE-RANGING CHANGES IN GENE EXPRESSION IN THE HIPPOCAMPUS. EPIGENETIC CHANGES TO DNA ARE ATTRACTIVE MECHANISMS TO EXPLAIN THE SUSTAINED HYPEREXCITABILITY OF CHRONIC EPILEPSY. HERE, THROUGH METHYLATION ANALYSIS OF ALL ANNOTATED C-PHOSPHATE-G ISLANDS AND PROMOTER REGIONS IN THE HUMAN GENOME, WE REPORT A PILOT STUDY OF THE METHYLATION PROFILES OF TEMPORAL LOBE EPILEPSY WITH OR WITHOUT HIPPOCAMPAL SCLEROSIS. FURTHERMORE, BY COMPARATIVE ANALYSIS OF EXPRESSION AND PROMOTER METHYLATION, WE IDENTIFY METHYLATION SENSITIVE NON-CODING RNA IN HUMAN TEMPORAL LOBE EPILEPSY. A TOTAL OF 146 PROTEIN-CODING GENES EXHIBITED ALTERED DNA METHYLATION IN TEMPORAL LOBE EPILEPSY HIPPOCAMPUS (N = 9) WHEN COMPARED TO CONTROL (N = 5), WITH 81.5% OF THE PROMOTERS OF THESE GENES DISPLAYING HYPERMETHYLATION. UNIQUE METHYLATION PROFILES WERE EVIDENT IN TEMPORAL LOBE EPILEPSY WITH OR WITHOUT HIPPOCAMPAL SCLEROSIS, IN ADDITION TO A COMMON METHYLATION PROFILE REGARDLESS OF PATHOLOGY GRADE. GENE ONTOLOGY TERMS ASSOCIATED WITH DEVELOPMENT, NEURON REMODELLING AND NEURON MATURATION WERE OVER-REPRESENTED IN THE METHYLATION PROFILE OF WATSON GRADE 1 SAMPLES (MILD HIPPOCAMPAL SCLEROSIS). IN ADDITION TO GENES ASSOCIATED WITH NEURONAL, NEUROTRANSMITTER/SYNAPTIC TRANSMISSION AND CELL DEATH FUNCTIONS, DIFFERENTIAL HYPERMETHYLATION OF GENES ASSOCIATED WITH TRANSCRIPTIONAL REGULATION WAS EVIDENT IN TEMPORAL LOBE EPILEPSY, BUT OVERALL FEW GENES PREVIOUSLY ASSOCIATED WITH EPILEPSY WERE AMONG THE DIFFERENTIALLY METHYLATED. FINALLY, A PANEL OF 13, METHYLATION-SENSITIVE MICRORNA WERE IDENTIFIED IN TEMPORAL LOBE EPILEPSY INCLUDING MIR27A, MIR-193A-5P (MIR193A) AND MIR-876-3P (MIR876), AND THE DIFFERENTIAL METHYLATION OF LONG NON-CODING RNA DOCUMENTED FOR THE FIRST TIME. THE PRESENT STUDY THEREFORE REPORTS SELECT, GENOME-WIDE DNA METHYLATION CHANGES IN HUMAN TEMPORAL LOBE EPILEPSY THAT MAY CONTRIBUTE TO THE MOLECULAR ARCHITECTURE OF THE EPILEPTIC BRAIN.	2015	
                                                                                                                                                                                                                                                                                                                                                 
4 2654  33 EPILEPSY PROGRESSION IS ASSOCIATED WITH CUMULATIVE DNA METHYLATION CHANGES IN INFLAMMATORY GENES. MESIAL TEMPORAL LOBE EPILEPSY WITH HIPPOCAMPAL SCLEROSIS (MTLE-HS) IS THE MOST COMMON FOCAL EPILEPSY IN ADULTS. IT IS CHARACTERIZED BY ALARMING RATES OF PHARMACORESISTANCE. EPILEPTOGENESIS IS ASSOCIATED WITH THE OCCURRENCE OF EPIGENETIC ALTERATIONS, AND THE FEW EPIGENETIC STUDIES CARRIED OUT IN MTLE-HS HAVE MAINLY FOCUSED ON THE HIPPOCAMPUS. IN THIS STUDY, WE OBTAINED THE DNA METHYLATION PROFILES FROM BOTH THE HIPPOCAMPUS AND ANTERIOR TEMPORAL NEOCORTEX OF MTLE-HS PATIENTS SUBJECTED TO RESECTIVE EPILEPSY SURGERY AND AUTOPSIED NON-EPILEPTIC CONTROLS. WE ASSESSED THE PROGRESSIVE NATURE OF DNA METHYLATION CHANGES IN RELATION TO EPILEPSY DURATION. WE IDENTIFIED SIGNIFICANTLY ALTERED HIPPOCAMPAL DNA METHYLATION PATTERNS ENCOMPASSING MULTIPLE PATHWAYS KNOWN TO BE INVOLVED IN EPILEPTOGENESIS. DNA METHYLATION CHANGES WERE EVEN MORE STRIKING IN THE NEOCORTEX, WHEREIN PATHOGENIC PATHWAYS AND GENES WERE COMMON TO BOTH TISSUES. MOST IMPORTANTLY, DNA METHYLATION CHANGES AT MANY GENOMIC SITES VARIED SIGNIFICANTLY WITH EPILEPSY DURATION. SUCH PROGRESSIVE CHANGES WERE ASSOCIATED WITH INFLAMMATION-RELATED GENES IN THE HIPPOCAMPUS. OUR RESULTS SUGGEST THAT THE NEOCORTEX, RELATIVELY SPARED OF EXTENSIVE HISTOPATHOLOGICAL DAMAGE, MAY ALSO BE INVOLVED IN EPILEPSY DEVELOPMENT. THESE RESULTS ALSO OPEN THE POSSIBILITY THAT THE OBSERVED NEOCORTICAL IMPAIRMENT COULD REPRESENT A PRELIMINARY STAGE OF EPILEPTOGENESIS BEFORE THE ESTABLISHMENT OF CHRONIC LESIONS OR A CONSEQUENCE OF PROLONGED SEIZURE EXPOSURE. OUR TWO-TISSUE MULTI-LEVEL CHARACTERIZATION OF THE MTLE-HS DNA METHYLOME SUGGESTS THE OCCURRENCE OF A SELF-PROPAGATING INFLAMMATORY WAVE OF EPIGENETIC DYSREGULATION.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
5 5273  38 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
6 5345  47 RAPID CHANGES IN EXPRESSION OF CLASS I AND IV HISTONE DEACETYLASES DURING EPILEPTOGENESIS IN MOUSE MODELS OF TEMPORAL LOBE EPILEPSY. A PROMINENT ROLE OF EPIGENETIC MECHANISMS IN MANIFESTATION OF EPILEPSY HAS BEEN PROPOSED. THUS ALTERED HISTONE H3 AND H4 ACETYLATION HAS BEEN DEMONSTRATED IN EXPERIMENTAL MODELS OF TEMPORAL LOBE EPILEPSY (TLE). WE NOW INVESTIGATED CHANGES IN THE EXPRESSION OF THE CLASS I AND CLASS IV HISTONE DEACETYLASES (HDAC) IN TWO COMPLEMENTARY MOUSE TLE MODELS. UNILATERAL INTRAHIPPOCAMPAL INJECTION OF KAINIC ACID (KA) INDUCED A STATUS EPILEPTICUS LASTING 6 TO 24H, DEVELOPMENT OF SPONTANEOUS LIMBIC SEIZURES (2 TO 3 DAYS AFTER KA INJECTION) AND CHRONIC EPILEPSY, AS REVEALED BY TELEMETRIC RECORDINGS OF THE EEGS. MICE WERE KILLED AT DIFFERENT INTERVALS AFTER KA INJECTION AND EXPRESSION OF HDAC MRNAS WAS INVESTIGATED BY IN SITU HYBRIDIZATION. WE OBSERVED MARKED DECREASES IN THE EXPRESSION OF HDACS 1, 2 AND 11 (BY UP TO 75%) IN THE GRANULE CELL AND PYRAMIDAL CELL LAYERS OF THE HIPPOCAMPUS DURING THE ACUTE STATUS EPILEPTICUS (2 TO 6H AFTER KA INJECTION). THIS WAS FOLLOWED BY INCREASED EXPRESSION OF ALL CLASS I HDAC MRNAS IN ALL PRINCIPAL CELL LAYERS OF THE HIPPOCAMPUS AFTER 12 TO 48 H. IN THE CHRONIC PHASE, 14 AND 28 DAYS AFTER KA, ONLY MODEST INCREASES IN THE EXPRESSION OF HDAC1 MRNA WERE OBSERVED IN GRANULE AND PYRAMIDAL CELLS. IMMUNOHISTOCHEMISTRY USING AN ANTIBODY DETECTING HDAC2 REVEALED RESULTS CONSISTENT WITH THE MRNA DATA AND INDICATES ALSO EXPRESSION IN GLIAL CELLS ON THE INJECTION SIDE. SIMILAR CHANGES AS SEEN IN THE KA MODEL WERE OBSERVED AFTER A PILOCARPINE-INDUCED STATUS EPILEPTICUS EXCEPT THAT DECREASES IN HDACS 2, 3 AND 8 WERE ALSO SEEN AT THE CHRONIC 28 DAY INTERVAL. THE PROMINENT DECREASES IN HDAC EXPRESSION DURING STATUS EPILEPTICUS ARE CONSISTENT WITH THE PREVIOUSLY DEMONSTRATED INCREASED EXPRESSION OF NUMEROUS PROTEINS AND WITH THE AUGMENTED ACETYLATION OF HISTONE H4. IT IS SUGGESTED THAT RESPECTIVE PUTATIVE GENE PRODUCTS COULD FACILITATE PROCONVULSIVE AS WELL AS ANTICONVULSIVE MECHANISMS. THE INCREASED EXPRESSION OF ALL CLASS I HDACS DURING THE "SILENT PHASE", ON THE OTHER HAND, MAY BE RELATED TO DECREASED HISTONE ACETYLATION, WHICH COULD CAUSE A DECREASE IN EXPRESSION OF CERTAIN PROTEINS, A MECHANISM THAT COULD ALSO PROMOTE EPILEPTOGENESIS. THUS, ADDRESSING HDAC EXPRESSION MAY HAVE A THERAPEUTIC POTENTIAL IN INTERFERING WITH A STATUS EPILEPTICUS AND WITH THE MANIFESTATION OF TLE.	2015	

7 5617  39 SARCOSINE SUPPRESSES EPILEPTOGENESIS IN RATS WITH EFFECTS ON HIPPOCAMPAL DNA METHYLATION. EPILEPTOGENESIS IS A COMMON CONSEQUENCE OF BRAIN INSULTS, HOWEVER, THE PREVENTION OR DELAY OF THE EPILEPTOGENIC PROCESS REMAINS AN IMPORTANT UNMET MEDICAL CHALLENGE. OVEREXPRESSION OF GLYCINE TRANSPORTER 1 (GLYT1) IS PROPOSED AS A PATHOLOGICAL HALLMARK IN THE HIPPOCAMPUS OF PATIENTS WITH TEMPORAL LOBE EPILEPSY (TLE), AND WE PREVIOUSLY DEMONSTRATED IN RODENT EPILEPSY MODELS THAT AUGMENTATION OF GLYCINE SUPPRESSED CHRONIC SEIZURES AND ALTERED ACUTE SEIZURE THRESHOLDS. IN THE PRESENT STUDY WE EVALUATED THE EFFECT OF THE GLYT1 INHIBITOR, SARCOSINE (AKA N-METHYLGLYCINE), ON EPILEPTOGENESIS AND ALSO INVESTIGATED POSSIBLE MECHANISMS. WE DEVELOPED A MODIFIED RAPID KINDLING MODEL OF EPILEPTOGENESIS IN RATS COMBINED WITH SEIZURE SCORE MONITORING TO EVALUATE THE ANTIEPILEPTOGENIC EFFECT OF SARCOSINE. WE USED IMMUNOHISTOCHEMISTRY AND WESTERN BLOT ANALYSIS FOR THE EVALUATION OF GLYT1 EXPRESSION AND EPIGENETIC CHANGES OF 5-METHYLCYTOSINE (5MC) AND 5-HYDROXYMETHYLCYTOSINE (5HMC) IN THE EPILEPTOGENIC HIPPOCAMPI OF RATS, AND FURTHER EVALUATED EXPRESSION CHANGES IN ENZYMES INVOLVED IN THE REGULATION OF DNA METHYLATION, TEN-ELEVEN TRANSLOCATION METHYLCYTOSINE DIOXYGENASE 1 (TET1), DNA-METHYLTRANSFERASE 1 (DNMT1), AND DNMT3A. OUR RESULTS DEMONSTRATED: (I) EXPERIMENTAL EVIDENCE THAT SARCOSINE (3 G/KG, I.P. DAILY) SUPPRESSED KINDLING EPILEPTOGENESIS IN RATS; (II) THE SARCOSINE-INDUCED ANTIEPILEPTOGENIC EFFECT WAS ACCOMPANIED BY A SUPPRESSED HIPPOCAMPAL GLYT1 EXPRESSION AS WELL AS A REDUCTION OF HIPPOCAMPAL 5MC LEVELS AND A CORRESPONDING INCREASE IN 5HMC; AND (III) SARCOSINE TREATMENT CAUSED DIFFERENTIAL EXPRESSION CHANGES OF TET1 AND DNMTS. TOGETHER, THESE FINDINGS SUGGEST THAT SARCOSINE HAS UNPRECEDENTED DISEASE-MODIFYING PROPERTIES IN A KINDLING MODEL OF EPILEPTOGENESIS IN RATS, WHICH WAS ASSOCIATED WITH ALTERED HIPPOCAMPAL DNA METHYLATION. THUS, MANIPULATION OF THE GLYCINE SYSTEM IS A POTENTIAL THERAPEUTIC APPROACH TO ATTENUATE THE DEVELOPMENT OF EPILEPSY.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                           
8  433  32 ANTIDEPRESSANT TREATMENT IS ASSOCIATED WITH EPIGENETIC ALTERATIONS OF HOMER1 PROMOTER IN A MOUSE MODEL OF CHRONIC DEPRESSION. BACKGROUND: UNDERSTANDING THE NEUROBIOLOGY OF DEPRESSION AND THE MECHANISM OF ACTION OF THERAPEUTIC MEASURES IS CURRENTLY A RESEARCH PRIORITY. WE HAVE SHOWN THAT THE EXPRESSION OF THE SYNAPTIC PROTEIN HOMER1A CORRELATES WITH DEPRESSION-LIKE BEHAVIOR AND ITS INDUCTION IS A COMMON MECHANISM OF ACTION OF DIFFERENT ANTIDEPRESSANT TREATMENTS. HOWEVER, THE MECHANISM OF HOMER1A REGULATION IS STILL UNKNOWN. METHODS: WE COMBINED THE CHRONIC DESPAIR MOUSE MODEL (CDM) OF CHRONIC DEPRESSION WITH DIFFERENT ANTIDEPRESSANT TREATMENTS. DEPRESSION-LIKE BEHAVIOR WAS CHARACTERIZED BY FORCED SWIM AND TAIL SUSPENSION TESTS, AND VIA AUTOMATIC MEASUREMENT OF SUCROSE PREFERENCE IN INTELLICAGE. THE HOMER1 MRNA EXPRESSION AND PROMOTER DNA METHYLATION WERE ANALYZED IN CORTEX AND PERIPHERAL BLOOD BY QRT-PCR AND PYROSEQUENCING. RESULTS: CDM MICE SHOW DECREASED HOMER1A AND HOMER1B/C MRNA EXPRESSION IN CORTEX AND BLOOD SAMPLES, WHILE CHRONIC TREATMENT WITH IMIPRAMINE AND FLUOXETINE OR ACUTE KETAMINE APPLICATION INCREASES THEIR LEVEL ONLY IN THE CORTEX. THE QUANTITATIVE ANALYSES OF THE METHYLATION OF 7 CPG SITES, LOCATED ON THE HOMER1 PROMOTER REGION CONTAINING SEVERAL CRE BINDING SITES, SHOW A SIGNIFICANT INCREASE IN DNA METHYLATION IN THE CORTEX OF CDM MICE. IN CONTRAST, ANTIDEPRESSANT TREATMENTS REDUCE THE METHYLATION LEVEL. LIMITATIONS: HOMER1 EXPRESSION AND PROMOTOR METHYLATION WERE NOT ANALYZED IN DIFFERENT BLOOD CELL TYPES. OTHER CPG SITES OF HOMER1 PROMOTER SHOULD BE INVESTIGATED IN FUTURE STUDIES. OUR EXPERIMENTAL APPROACH DOES NOT DISTINGUISH BETWEEN METHYLATION AND HYDROXYMETHYLATION. CONCLUSIONS: WE DEMONSTRATE THAT STRESS-INDUCED DEPRESSION-LIKE BEHAVIOR AND ANTIDEPRESSANT TREATMENTS ARE ASSOCIATED WITH EPIGENETIC ALTERATIONS OF HOMER1 PROMOTER, PROVIDING NEW INSIGHTS INTO THE MECHANISM OF ANTIDEPRESSANT TREATMENT.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
9 6411  34 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	
                                                  
10 5205  32 PRENATAL STRESS CHANGES THE GLYCOPROTEIN GPM6A GENE EXPRESSION AND INDUCES EPIGENETIC CHANGES IN RAT OFFSPRING BRAIN. PRENATAL STRESS (PS) EXERTS STRONG IMPACT ON FETAL BRAIN DEVELOPMENT AND ON ADULT OFFSPRING BRAIN FUNCTIONS. PREVIOUS WORK DEMONSTRATED THAT CHRONIC STRESS ALTERS THE MRNA EXPRESSION OF GPM6A, A NEURONAL GLYCOPROTEIN INVOLVED IN FILOPODIUM EXTENSION. IN THIS WORK, WE ANALYZED THE EFFECT OF PS ON GPM6A EXPRESSION AND THE EPIGENETIC MECHANISMS INVOLVED. PREGNANT WISTAR RATS RECEIVED RESTRAINT STRESS DURING THE LAST WEEK OF GESTATION. MALE OFFSPRING WERE SACRIFICED ON POSTNATAL DAYS 28 AND 60. HIPPOCAMPUS AND PREFRONTAL CORTEX SAMPLES WERE ANALYZED FOR GENE EXPRESSION (QPCR FOR MRNAS AND MICRORNAS), METHYLATION STATUS (BISULFITE CONVERSION) AND PROTEIN LEVELS. HIPPOCAMPAL NEURONS IN CULTURE WERE USED TO ANALYZE MICRORNA OVEREXPRESSION EFFECTS. PRENATAL STRESS INDUCED CHANGES IN GPM6A LEVELS IN BOTH TISSUES AND AT BOTH AGES ANALYZED, INDICATING A PERSISTENT EFFECT. TWO CPG ISLANDS IN THE GPM6A GENE WERE IDENTIFIED. VARIATIONS IN THE METHYLATION PATTERN AT THREE SPECIFIC CPGS WERE FOUND IN HIPPOCAMPUS, BUT NOT IN PFC SAMPLES FROM PS OFFSPRING. MICRORNAS PREDICTED TO TARGET GPM6A WERE IDENTIFIED IN SILICO. QPCR MEASUREMENTS SHOWED THAT PS MODIFIED THE EXPRESSION OF SEVERAL MICRORNAS IN BOTH TISSUES, BEING MICRORNA-133B THE MOST SIGNIFICANTLY ALTERED. FURTHER STUDIES OVEREXPRESSING THIS MICRORNA IN NEURONAL CULTURES SHOWED A REDUCTION IN GMP6A MRNA AND PROTEIN LEVEL. MOREOVER FILOPODIUM DENSITY WAS ALSO REDUCED, SUGGESTING THAT GPM6A FUNCTION WAS AFFECTED. GESTATIONAL STRESS AFFECTED GPM6A GENE EXPRESSION IN OFFSPRING LIKELY THROUGH CHANGES IN METHYLATION STATUS AND IN POSTTRANSCRIPTIONAL REGULATION BY MICRORNAS. THUS, OUR FINDINGS PROPOSE GPM6A AS A NOVEL TARGET FOR EPIGENETIC REGULATION DURING PRENATAL STRESS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
11 4221  41 METHYLATION AND SILENCING OF PROTEIN TYROSINE PHOSPHATASE RECEPTOR TYPE O IN CHRONIC LYMPHOCYTIC LEUKEMIA. PURPOSE: PREVIOUS STUDIES IN OUR LABORATORY HAVE SHOWN THE PROGRESSIVE METHYLATION AND SUPPRESSION OF THE GENE ENCODING PROTEIN TYROSINE PHOSPHATASE, PTPRO, IN THE LIVERS OF RATS FED A METHYL-DEFICIENT DIET THAT INDUCES HEPATOCARCINOGENESIS. SUBSEQUENTLY, WE OBSERVED THE METHYLATION OF PTPRO IN PRIMARY HUMAN LUNG TUMORS AND ALSO SHOWED ITS POTENTIAL TUMOR SUPPRESSOR CHARACTERISTICS. THE PRESENT STUDY WAS UNDERTAKEN TO INVESTIGATE WHETHER THE TRUNCATED FORM OF PTPRO (PTPROT), SPECIFICALLY EXPRESSED IN NAIVE B LYMPHOCYTES, WAS ALSO METHYLATED AND SUPPRESSED IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL), A DISEASE GENERALLY AFFECTING B LYMPHOCYTES. EXPERIMENTAL DESIGN AND RESULTS: INITIAL SCREENING SHOWED THAT 60% OF THE 52 CLL SAMPLES ANALYZED USING METHYLATION-SPECIFIC PCR ASSAY WERE METHYLATED COMPARED WITH B LYMPHOCYTES FROM NORMAL INDIVIDUALS, WHICH WERE NOT METHYLATED. THE EXPRESSION OF PTPROT, AS MEASURED BY SEMIQUANTITATIVE REVERSE TRANSCRIPTION-PCR, INVERSELY CORRELATED WITH METHYLATION IN THE FEW SAMPLES TESTED. ANALYSIS OF ADDITIONAL SAMPLES (N = 50) BY COMBINED BISULFITE RESTRICTION ANALYSIS SHOWED THAT THE PTPRO CPG ISLAND WAS METHYLATED IN 82% OF PATIENTS WITH CLL COMPARED WITH B LYMPHOCYTES FROM NORMAL INDIVIDUALS. FURTHERMORE, OVERALL EXPRESSION OF PTPRO WAS REDUCED IN CLL RELATIVE TO NORMAL LYMPHOCYTES. THE PTPRO GENE WAS ALSO SUPPRESSED BY METHYLATION IN THE CLL CELL LINE WAC3CD5, WHERE IT COULD BE REACTIVATED UPON TREATMENT WITH THE DNA HYPOMETHYLATING AGENT 5-AZAC. ECTOPIC EXPRESSION OF PTPROT IN A NONEXPRESSING CELL LINE INCREASED GROWTH INHIBITION WITH FLUDARABINE TREATMENT, A THERAPY COMMONLY USED FOR CLL. CONCLUSION: THIS STUDY REVEALS THE POTENTIAL ROLE OF PTPRO METHYLATION AND SILENCING IN CLL TUMORIGENESIS AND ALSO PROVIDES A NOVEL MOLECULAR TARGET IN THE EPIGENETIC THERAPY.	2007	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
12 3647  31 INCREASED REELIN PROMOTER METHYLATION IS ASSOCIATED WITH GRANULE CELL DISPERSION IN HUMAN TEMPORAL LOBE EPILEPSY. MESIAL TEMPORAL SCLEROSIS (MTS) IS THE MOST COMMON LESION IN CHRONIC, INTRACTABLE TEMPORAL LOBE EPILEPSIES (TLE) AND CHARACTERIZED BY SEGMENTAL NEURONAL CELL LOSS IN MAJOR HIPPOCAMPAL SEGMENTS. ANOTHER HISTOPATHOLOGICAL HALLMARK INCLUDES GRANULE CELL DISPERSION (GCD), AN ARCHITECTURAL DISTURBANCE OF THE DENTATE GYRUS ENCOUNTERED IN APPROXIMATELY 50% OF PATIENTS WITH MESIAL TEMPORAL SCLEROSIS. REELIN, WHICH PLAYS A KEY ROLE DURING HIPPOCAMPAL DEVELOPMENT AND MAINTENANCE OF LAMINAR ORGANIZATION, IS SYNTHESIZED AND RELEASED BY CAJAL-RETZIUS CELLS OF THE DENTATE MOLECULAR LAYER, AND PREVIOUS STUDIES HAVE SHOWN THAT REELIN TRANSCRIPT LEVELS ARE DOWNREGULATED IN HUMAN TEMPORAL LOBE EPILEPSIES SPECIMENS. TO INVESTIGATE WHETHER EPIGENETIC SILENCING BY REELIN PROMOTER METHYLATION MAY BE AN UNDERLYING PATHOGENETIC MECHANISM OF GCD, DNA WAS HARVESTED FROM 3 MICRODISSECTED HIPPOCAMPAL SUBREGIONS (I.E. MOLECULAR AND GRANULE CELL LAYERS OF THE DENTATE GYRUS AND PRESUBICULUM) FROM 8 MTS SPECIMENS WITH GCD, 5 TLE SAMPLES WITHOUT GCD, AND 3 AUTOPSY CONTROLS. PROMOTER METHYLATION WAS ANALYZED AFTER BISULFITE TREATMENT, CLONING, AND DIRECT SEQUENCING; IMMUNOHISTOCHEMISTRY WAS PERFORMED TO IDENTIFY CAJAL-RETZIUS CELLS. REELIN PROMOTER METHYLATION WAS FOUND TO BE GREATER IN TLE SPECIMENS THAN IN CONTROLS; PROMOTER METHYLATION CORRELATED WITH GCD AMONG TLE SPECIMENS (P < 0.0002). NO OTHER CLINICAL OR HISTOPATHOLOGICAL PARAMETER (I.E. SEX, AGE, SEIZURE DURATION, MEDICATION OR EXTENT, OF MTS) CORRELATED WITH PROMOTER METHYLATION. THESE DATA SUPPORT A COMPROMISED REELIN-SIGNALING PATHWAY AND IDENTIFY PROMOTER METHYLATION AS AN EPIGENETIC MECHANISM IN THE PATHOGENESIS OF TLE.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
13 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	
                                                                                                                                                                                                                                                                                                
14 2057  35 EPIGENETIC CONTROL OF EPILEPSY TARGET GENES CONTRIBUTES TO A CELLULAR MEMORY OF EPILEPTOGENESIS IN CULTURED RAT HIPPOCAMPAL NEURONS. HYPERSYNCHRONOUS NEURONAL EXCITATION MANIFESTS CLINICALLY AS SEIZURE (ICTOGENESIS), AND MAY RECUR SPONTANEOUSLY AND REPETITIVELY AFTER A VARIABLE LATENCY PERIOD (EPILEPTOGENESIS). DESPITE TREMENDOUS RESEARCH EFFORTS TO DESCRIBE MOLECULAR PATHWAYS AND SIGNATURES OF EPILEPTOGENESIS, MOLECULAR PATHOMECHANISMS LEADING TO CHRONIC EPILEPSY REMAIN TO BE CLARIFIED. WE HYPOTHESIZED THAT EPIGENETIC MODIFICATIONS MAY FORM THE BASIS FOR A CELLULAR MEMORY OF EPILEPTOGENESIS, AND USED A PRIMARY NEURONAL CELL CULTURE MODEL OF THE RAT HIPPOCAMPUS TO STUDY THE TRANSLATION OF MASSIVE NEURONAL EXCITATION INTO PERSISTING CHANGES OF EPIGENETIC SIGNATURES AND PRO-EPILEPTOGENIC TARGET GENE EXPRESSION. INCREASED SPONTANEOUS ACTIVATION OF CULTURED NEURONS WAS DETECTED 3 AND 7 DAYS AFTER STIMULATION WITH 10 MUM GLUTAMATE WHEN COMPARED TO SHAM-TREATED TIME-MATCHED CONTROLS USING CALCIUM-IMAGING IN VITRO. CHROMATIN-IMMUNOPRECIPITATION EXPERIMENTS REVEALED SHORT-TERM (3 H, 7 H, AND 24 H) AND LONG-TERM (3 D AND 2 WEEKS) CHANGES IN HISTONE MODIFICATIONS, WHICH WERE DIRECTLY LINKED TO DECREASED EXPRESSION OF TWO SELECTED EPILEPSY TARGET GENES, E.G. EXCITATORY GLUTAMATE RECEPTOR GENES GRIA2 AND GRIN2A. INCREASED PROMOTER METHYLATION OBSERVED 4 WEEKS AFTER GLUTAMATE STIMULATION AT RESPECTIVE GENES SUGGESTED LONG-TERM REPRESSION OF GRIA2 AND GRIN2A GENES. INHIBITION OF GLUTAMATERGIC ACTIVATION OR BLOCKING THE PROPAGATION OF ACTION POTENTIALS IN CULTURED NEURONS RESCUED ALTERED GENE EXPRESSION AND REGULATORY EPIGENETIC MODIFICATIONS. OUR DATA SUPPORT THE CONCEPT OF A CELLULAR MEMORY OF EPILEPTOGENESIS AND PERSISTING EPIGENETIC MODIFICATIONS OF EPILEPSY TARGET GENES, WHICH ARE ABLE TO TURN NORMAL INTO PRO-EPILEPTIC NEURONS AND CIRCUITS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
15 3494  37 IDENTIFICATION OF MICRORNAS WITH DYSREGULATED EXPRESSION IN STATUS EPILEPTICUS INDUCED EPILEPTOGENESIS. THE INVOLVEMENT OF MIRNA IN MESIAL TEMPORAL LOBE EPILEPSY (MTLE) PATHOGENESIS HAS INCREASINGLY BECOME A FOCUS OF EPIGENETIC STUDIES. DESPITE ADVANCES, THE NUMBER OF KNOWN MIRNAS WITH A CONSISTENT EXPRESSION RESPONSE DURING EPILEPTOGENESIS IS STILL SMALL. ADDRESSING THIS SITUATION REQUIRES ADDITIONAL MIRNA PROFILING STUDIES COUPLED TO DETAILED INDIVIDUAL EXPRESSION ANALYSES. HERE, WE PERFORM A MIRNA MICROARRAY ANALYSIS OF THE HIPPOCAMPUS OF WISTAR RATS 24 HOURS AFTER INTRA-HIPPOCAMPAL PILOCARPINE-INDUCED STATUS EPILEPTICUS (H-PILO SE). WE IDENTIFIED 73 MIRNAS THAT UNDERGO SIGNIFICANT CHANGES, OF WHICH 36 WERE UP-REGULATED AND 37 WERE DOWN-REGULATED. TO VALIDATE, WE SELECTED 5 OF THESE (10A-5P, 128A-3P, 196B-5P, 352 AND 324-3P) FOR RT-QPCR ANALYSIS. OUR RESULTS CONFIRMED THAT MIR-352 AND 196B-5P LEVELS WERE SIGNIFICANTLY HIGHER AND MIR-128A-3P LEVELS WERE SIGNIFICANTLY LOWER IN THE HIPPOCAMPUS OF H-PILO SE RATS. WE ALSO EVALUATED WHETHER THE 3 MIRNAS SHOW A DYSREGULATED HIPPOCAMPAL EXPRESSION AT THREE TIME PERIODS (0H, 24H AND CHRONIC PHASE) AFTER SYSTEMIC PILOCARPINE-INDUCED STATUS EPILEPTICUS (S-PILO SE). WE DEMONSTRATE THAT MIR-128A-3P TRANSCRIPTS ARE SIGNIFICANTLY REDUCED AT ALL TIME POINTS COMPARED TO THE NAIVE GROUP. MOREOVER, MIR-196B-5P WAS SIGNIFICANTLY HIGHER ONLY AT 24H POST-SE, WHILE MIR-352 TRANSCRIPTS WERE SIGNIFICANTLY UP-REGULATED AFTER 24H AND IN CHRONIC PHASE (EPILEPTIC) RATS. FINALLY, WHEN WE COMPARED HIPPOCAMPI OF EPILEPTIC AND NON-EPILEPTIC HUMANS, WE OBSERVED THAT TRANSCRIPT LEVELS OF MIRNAS SHOW SIMILAR TRENDS TO THE ANIMAL MODELS. IN SUMMARY, WE SUCCESSFULLY IDENTIFIED TWO NOVEL DYSREGULATED MIRNAS (196B-5P AND 352) AND CONFIRMED MIR-128A-3P DOWNREGULATION IN SE-INDUCED EPILEPTOGENESIS. FURTHER FUNCTIONAL ASSAYS ARE REQUIRED TO UNDERSTAND THE ROLE OF THESE MIRNAS IN MTLE PATHOGENESIS.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
16 1328  36 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
17 1297  35 DECREASED MRNA EXPRESSION LEVELS OF DNA METHYLTRANSFERASES TYPE 1 AND 3A IN SYSTEMIC LUPUS ERYTHEMATOSUS. OBJECTIVES: SYSTEMIC LUPUS ERYTHEMATOSUS (SLE) IS A CHRONIC RELAPSING AUTOIMMUNE DISEASE CHARACTERIZED BY THE PRESENCE OF AUTOANTIBODIES DIRECTED AGAINST NUCLEAR ANTIGENS AND BY CHRONIC INFLAMMATION. ALTHOUGH THE ETIOLOGY OF SLE REMAINS UNCLEAR, THE INFLUENCE OF ENVIRONMENT FACTORS, WHICH IS LARGELY REFLECTED BY THE EPIGENETIC MECHANISMS, WITH DNA METHYLATION CHANGES IN PARTICULAR, IS GENERALLY CONSIDERED AS MAIN PLAYERS IN THE PATHOGENESIS OF SLE. WE STUDIED DNA METHYLTRANSFERASES' (DNMTS) TYPE 1, 3A AND 3B TRANSCRIPT LEVELS IN PERIPHERAL BLOOD MONONUCLEAR CELLS FROM PATIENTS DIAGNOSED WITH SYSTEMIC LUPUS ERYTHEMATOSUS AND FROM THE HEALTHY CONTROL SUBJECTS. FURTHERMORE, THE ASSOCIATION OF DNMT1, DNMT3A, AND DNMT3B MRNA LEVELS WITH GENDER, AGE, AND MAJOR CLINICAL MANIFESTATIONS WAS ANALYZED. METHODS: PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) WERE ISOLATED FROM 32 SLE PATIENTS AND 40 HEALTHY CONTROLS. REVERSE TRANSCRIPTION AND REAL-TIME QUANTITATIVE POLYMERASE CHAIN REACTION (RT-QPCR) ANALYSES WERE USED TO DETERMINE DNMT1, DNMT3A, AND DNMT3B MRNA EXPRESSION LEVELS. RESULTS: SIGNIFICANTLY LOWER DNMT1 (P = 0.015543) AND DNMT3A (P = 0.003652) TRANSCRIPT LEVELS IN SLE PATIENTS WERE OBSERVED COMPARED WITH HEALTHY CONTROLS. NEVERTHELESS, THE DNMT3B MRNA EXPRESSION LEVELS WERE MARKEDLY LOWER COMPARED WITH DNMT1 AND DNMT3A, BOTH IN PBMCS FROM AFFECTED PATIENTS AND THOSE FROM CONTROL SUBJECTS. FURTHERMORE, THE DNMT1 TRANSCRIPT LEVELS WERE POSITIVELY CORRELATED WITH SLE DISEASE ACTIVITY INDEX (SLEDAI) (R (S) = 0.4087, P = 0.020224), WHILE THE DNMT3A TRANSCRIPT LEVELS WERE NEGATIVELY CORRELATED WITH PATIENTS AGE (R (S) = -0.3765, P = 0.03369). CONCLUSIONS: OUR ANALYSES CONFIRMED THE IMPORTANCE OF EPIGENETIC ALTERATIONS IN SLE ETIOLOGY. MOREOVER, OUR RESULTS SUGGEST THAT THE PRESENCE OF SOME CLINICAL MANIFESTATIONS, SUCH AS PHOTOTOSENSITIVITY AND ARTHRITIS, MIGHT BE ASSOCIATED WITH THE DYSREGULATION OF DNA METHYLTRANSFERASES' MRNA EXPRESSION LEVELS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                          
18 3986  43 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
19 2297  33 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
20  977  43 CHRONIC ORAL EXPOSURE TO INORGANIC ARSENATE INTERFERES WITH METHYLATION STATUS OF P16INK4A AND RASSF1A AND INDUCES LUNG CANCER IN A/J MICE. ALTHOUGH INORGANIC ARSENATE (IAS(V)) OR ARSENITE (IAS(III)) IS CLEARLY A HUMAN CARCINOGEN, IT HAS BEEN DIFFICULT TO PRODUCE TUMORS IN RODENTS. IN THE PRESENT STUDY, WE ORALLY ADMINISTERED IAS(V) TO A/J MICE TO EXAMINE ARSENIC CARCINOGENICITY IN RODENT. A/J MICE (MALE, N = 120) ASSIGNED TO FOUR GROUPS WERE GIVEN DRINKING WATER CONTAINING 0, 1, 10, AND 100 PPM IAS(V) FOR 18 MONTHS. AT THE END OF EXPERIMENT, THE COMPLETE LUNGS WERE REMOVED AND USED FOR EXAMINING HISTOPATHOLOGY AND EXTRACTING RNA AND DNA. EPIGENETIC EFFECTS OF IAS(V) ON DNA METHYLATION PATTERNS OF P16INK4A AND RASSF1A GENES WERE DETERMINED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. CHANGES OF P16INK4A AND RASSF1A AT MRNA AND PROTEIN LEVELS WERE EXAMINED BY REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION AND IMMUNOHISTOCHEMISTRY. ARSENIC WAS ACCUMULATED DOSE DEPENDENTLY IN THE LUNG TISSUES OF IAS(V)-EXPOSED MICE. INCREASE IN LUNG TUMOR NUMBER AND LUNG TUMOR SIZE WAS OBSERVED IN IAS(V)-EXPOSED MICE COMPARED TO THE CONTROL. HISTOPATHOLOGICAL EXAMINATION SHOWED THAT THE RATE OF POORLY DIFFERENTIATED LUNG ADENOCARCINOMA WAS MUCH HIGHER IN IAS(V)-EXPOSED MICE THAN IN THE CONTROL. METHYLATION RATES APPEARED TO BE HIGHER IN A DOSE-RELATED TENDENCY IN LUNG TUMORS FROM IAS(V)-EXPOSED MICE COMPARED TO THE CONTROL. LOWER OR LOSS OF P16INK4A AND RASSF1A EXPRESSION WAS FOUND IN LUNG TUMORS FROM IAS(V)-EXPOSED MICE, COMPARED TO THAT IN NONTUMOR LUNG TISSUES FROM BOTH CONTROL AND IAS(V)-EXPOSED MICE, AND THIS REDUCED OR LOST EXPRESSION WAS IN ACCORDANCE WITH HYPERMETHYLATION OF THE GENES. IN CONCLUSION, IAS(V) EXPOSURE INCREASED LUNG TUMOR INCIDENCE AND MULTIPLICITY IN A/J MICE. EPIGENETIC CHANGES OF TUMOR SUPPRESSOR GENES SUCH AS P16INK4A AND RASSF1A ARE INVOLVED IN THE IAS(V)-INDUCED LUNG CARCINOGENESIS.	2006