1 411 136 ANALYSIS OF GENOME-WIDE DNA METHYLATION PATTERNS IN OBESITY. OBESITY IS A CHRONIC AND COMPLEX PSYCHOSOMATIC DISEASE THAT IS BECOMING INCREASINGLY PREVALENT WORLDWIDE. THIS STUDY AIMED TO ANALYZE WHOLE METHYLATION PROFILES TO UNCOVER THE EPIGENETIC MECHANISMS ASSOCIATED WITH OBESITY. DNA METHYLATION PROFILES IN BLOOD SAMPLES FROM PATIENTS WITH OBESITY AND NORMAL CONTROLS WERE STUDIED USING THE ILLUMINA 850 K METHYLATION MICROARRAY. THE DIAGNOSTIC VALUE OF THE DIFFERENTIALLY METHYLATED GENES WAS DETERMINED USING RECEIVER OPERATING CHARACTERISTIC (ROC) ANALYSIS. THE EXPRESSION OF SELECTED CANDIDATE GENES WAS VERIFIED USING REVERSE TRANSCRIPTION QUANTITATIVE POLYMERASE CHAIN REACTION (RT-QPCR) AND PYROSEQUENCING. A TOTAL OF 9,371 SIGNIFICANTLY DIFFERENTIALLY METHYLATED SITES (7,974 HYPERMETHYLATED SITES AND 1,397 HYPOMETHYLATED SITES) WERE IDENTIFIED IN 4,571 GENES. A DIFFERENCE IN THE DISTRIBUTION OF DIFFERENTIALLY METHYLATED SITES (HYPERMETHYLATED AND HYPOMETHYLATED) IN BOTH GENE STRUCTURES AND CPG ISLANDS WAS OBSERVED. A TOTAL OF 114 KEY DIFFERENTIALLY METHYLATED SITES WERE IDENTIFIED IN THE CPG ISLANDS. ROC RESULTS INDICATED THAT INHIBIN SUBUNIT BETA B (INHBB), HOMEOBOX A9 (HOXA9), TROPONIN T3 (TNNT3), CYCLIC ADENOSINE MONOPHOSPHATE (CAMP)-RESPONSIVE ELEMENT BINDING PROTEIN (CREB)-REGULATED TRANSCRIPTION COACTIVATOR 1 (CRTC1) AND ZINC FINGER AND BTB DOMAIN-CONTAINING 7 B (ZBTB7B) COULD DISCRIMINATE PATIENTS WITH OBESITY FROM NORMAL CONTROLS. RT-QPCR RESULTS OF CRTC1 AND ZBTB7B WERE CONSISTENT WITH OUR METHYLATION PROFILE RESULTS. THE PYROSEQUENCING RESULTS SHOWED THAT THE METHYLATION LEVELS OF CRTC1 CPG SITES (CPG1 AND CPG2-CG11660071) AND INHBB CPG SITES (CPG2) WERE SIGNIFICANTLY CHANGED IN PATIENTS WITH OBESITY COMPARED WITH NORMAL CONTROLS, WHICH WAS CONSISTENT WITH OUR DNA METHYLATION PROFILE RESULTS. OUR STUDY PROVIDES NEW INSIGHTS INTO THE PATHOLOGICAL MECHANISM OF OBESITY. 2021 2 1994 29 EPIGENETIC AND GENE EXPRESSION ALTERATIONS OF FOXP3 IN THE T CELLS OF EAE MOUSE MODEL OF MULTIPLE SCLEROSIS. MULTIPLE SCLEROSIS (MS) IS A CHRONIC AUTOIMMUNE DISEASE WITH DEMYELINATION AND NEURODEGENERATION OF THE CENTRAL NERVOUS SYSTEM. IT HAS BEEN SHOWN THAT THE REGULATORY T (TREG) CELLS ARE RESPONSIBLE FOR MAINTAINING TOLERANCE TO SELF-ANTIGENS AND CAN SUPPRESS THE AUTOIMMUNE PROCESS IN SEVERAL ANIMAL MODELS SUCH AS EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE), A MOUSE MODEL OF MS. RECENT BASIC STUDIES HAVE DEMONSTRATED THAT FORKHEAD BOX P (FOXP3) AND BTB DOMAIN AND CNC HOMOLOG 2 (BACH2) ARE THE MASTER TRANSCRIPTION FACTORS OF THESE CELLS PLAYING A PIVOTAL ROLE IN THE POLARIZATION OF NAIVE T CELLS INTO TREG CELLS. IN THE CURRENT STUDY, THE EXPRESSION OF FOXP3 AND BACH2 GENES AND FOXP3 PROMOTER METHYLATION WERE EVALUATED IN T CELLS OF THE EAE-INDUCED MICE. THE RESULTS OF THIS STUDY SHOWED A PROMINENT AND SIGNIFICANT HYPERMETHYLATION OF THE FOXP3 GENE PROMOTER IN THE EAE-INDUCED MICE COMPARED TO THE SHAM AND CONTROL GROUPS. THE EXPRESSION OF FOXP3 AND BACH2 GENES WAS SIGNIFICANTLY DECREASED IN THE EAE GROUP IN COMPARISON WITH THE SHAM AND CONTROL GROUPS. THIS STUDY SUGGESTS THAT THE EPIGENETIC MODIFICATION OF FOXP3 GENE IS INVOLVED IN THE PATHOGENESIS OF EAE AND THIS COULD BE IMPORTANT IN THERAPY IN AN APPROPRIATE AND LOGICAL STATEMENT. 2017 3 5348 37 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 4 3448 36 HYPERMETHYLATION OF THE N-MYC DOWNSTREAM-REGULATED GENE 2 PROMOTER IN PERIPHERAL BLOOD MONONUCLEAR CELLS IS ASSOCIATED WITH LIVER FIBROSIS IN CHRONIC HEPATITIS B. DNA METHYLATION IS A FUNDAMENTAL EPIGENETIC MODIFICATION TO REGULATE GENE EXPRESSION. N-MYC DOWNSTREAM-REGULATED GENE (NDRG) 2 IS A CYTOPLASMIC PROTEIN AND PARTICIPATES IN THE PATHOGENESIS OF LIVER FIBROSIS. IN THIS STUDY, THE MRNA EXPRESSION AND METHYLATION STATUS OF NDRG2 WAS EVALUATED IN PATIENTS WITH CHRONIC HEPATITIS B (CHB). THE STUDY INCLUDED 143 CHB PATIENTS AND 65 NORMAL CONTROLS (NC). THE MRNA EXPRESSION OF NDRG2 IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) WAS DETECTED BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION. THE METHYLATION STATUS OF THE NDRG2 PROMOTER IN PBMCS WAS DETECTED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. THE NDRG2 MRNA LEVEL WAS LOWER IN THE CHB GROUP THAN IN THE NC GROUP (P < 0.001). METHYLATION FREQUENCY OF THE NDRG2 PROMOTER WAS SIGNIFICANTLY HIGHER IN CHB PATIENTS THAN IN THE NC GROUP (52.44% VS. 26.15%, P < 0.001). IMPORTANTLY, THE RELATIVE EXPRESSION LEVELS OF NDRG2 MRNA WERE SIGNIFICANTLY LOWER IN THE METHYLATED GROUP THAN IN THE UNMETHYLATED GROUP IN BOTH CHB PATIENTS AND NC (P < 0.001). FURTHERMORE, A LOWER MRNA LEVEL AND HYPERMETHYLATION OF NDRG2 WERE ASSOCIATED WITH LIVER FIBROSIS AND INFLAMMATION GRADE IN CHB. THE ASPARTATE AMINOTRANSFERASE-TO-PLATELET RATIO INDEX (APRI) SCORE IS WIDELY USED TO PREDICT LIVER FIBROSIS. THE MRNA EXPRESSION LEVELS AND METHYLATION STATUS OF NDRG2 SHOWED A BETTER SCORE COMPARED TO APRI FOR DISCRIMINATING THE SEVERITY OF LIVER FIBROSIS. IN CONCLUSION, HYPERMETHYLATION OF NDRG2 IN PBMCS WAS CORRELATED WITH DECREASED MRNA EXPRESSION AND WITH LIVER FIBROSIS. THE METHYLATION STATUS OF THE NDRG2 PROMOTER IN PBMCS IS A POTENTIAL NONINVASIVE BIOMARKER TO PREDICT THE SEVERITY OF LIVER FIBROSIS. 2017 5 5553 48 ROLE OF EPIGENETICS IN THE PATHOGENESIS OF CHRONIC RHINOSINUSITIS WITH NASAL POLYPS. CHRONIC RHINOSINUSITIS (CRS) IS A HIGHLY PREVALENT DISEASE CHARACTERIZED BY MUCOSAL INFLAMMATION OF THE NOSE AND PARANASAL SINUSES. CRS CAN BE DIVIDED INTO TWO MAIN CATEGORIES, CRS WITH NASAL POLYPS (NPS; CRSWNP) AND CRS WITHOUT NPS (CRSSNP). ALTHOUGH THE PATHOPHYSIOLOGY OF CRS REMAINS UNCLEAR, DNA METHYLATION HAS BEEN IMPLICATED IN THE ETIOLOGY OF CRSWNP. THE AIM OF THE PRESENT STUDY WAS TO ELUCIDATE WHETHER DNA METHYLATION OF SPECIFIC GENES IS INVOLVED IN THE DEVELOPMENT OF NPS. IN TOTAL, 18 INDIVIDUALS WERE INCLUDED IN THE PRESENT STUDY, AND WERE DIVIDED INTO THREE GROUPS: CRSWNP (N=7), CRSSNP (N=7) AND HEALTHY CONTROLS (N=4). NP TISSUES WERE OBTAINED FROM THE SEVEN PATIENTS WITH CRSWNP AND BIOPSIES OF THE INFERIOR TURBINATE MUCOSA FROM ALL THREE GROUPS WERE USED AS CONTROLS. METHYLATED GENES DETECTED BY METHYL?CPG?BINDING DOMAIN SEQUENCING WERE VALIDATED BY METHYLATION?SPECIFIC POLYMERASE CHAIN REACTION (PCR), BISULFITE SEQUENCING, AND REVERSE TRANSCRIPTION?QUANTITATIVE PCR (RT?QPCR). METHYL?CPG?BINDING DOMAIN SEQUENCING IDENTIFIED 43,674 CPG ISLANDS IN 518 GENES. THE PROMOTOR REGIONS OF 10 AND 30 GENES WERE HYPERMETHYLATED AND HYPOMETHYLATED, RESPECTIVELY, IN NP SAMPLES COMPARED WITH CONTROLS. THE TOP FOUR GENES WITH ALTERED HYPOMETHYLATION IN NP TISSUES WERE, KERATIN 19 (KRT19), NUCLEAR RECEPTOR SUBFAMILY 2 GROUP F MEMBER 2 (NR2F2), A DISINTEGRIN?LIKE AND METALLOPEPTIDASE (REPROLYSIN TYPE) WITH THROMBOSPONDIN TYPE 1 MOTIF 1 (ADAMTS1) AND ZINC FINGER PROTEIN 222 (ZNF222). RT?QPCR DEMONSTRATED THAT THE EXPRESSION LEVELS OF KRT19, NR2F2 AND ADAMTS1 WERE SIGNIFICANTLY INCREASED IN NP TISSUES; HOWEVER, THERE WAS NO DIFFERENCE IN THE LEVELS OF ZNF222 BETWEEN NP AND CONTROL TISSUES. FURTHER STUDIES ARE REQUIRED TO CONFIRM THE RELEVANCE OF THESE EPIGENETIC MODIFICATIONS IN THE MECHANISMS UNDERLYING NP FORMATION. 2018 6 4601 35 NDRG2 MRNA LEVELS AND MIR-28-5P AND MIR-650 ACTIVITY IN CHRONIC LYMPHOCYTIC LEUKEMIA. BACKGROUND: NDRG2 IS IDENTIFIED AS A TUMOR SUPPRESSOR GENE IN MANY TUMORS, AND FUNCTIONS IN CELL PROLIFERATION, DIFFERENTIATION AND APOPTOSIS. RECENT DATA INDICATE THAT NDRG2 EXPRESSION IS UP-REGULATED BY TP53. MOREOVER, PROPOSED MECHANISMS OF NDRG2 INACTIVATION INCLUDE EPIGENETIC SILENCING OF THE NDRG2 PROMOTER AND DOWN-REGULATION BY MICRORNAS (MIRNAS). HOWEVER, FEW STUDIES HAVE EVER BEEN DONE ON THE ROLE OF NDRG2 AND THE NDRG2-REGULATING MIRNAS INTERFERENCE IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). METHODS: NDRG2 AND MICRORNAS MRNA LEVELS IN CLL SUBJECTS WERE ASSESSED BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QRT-PCR). THE DUAL-LUCIFERASE REPORTER ASSAY WAS PERFORMED TO DETERMINE NDRG2-RELATED MIRNAS. LOW EXPRESSION OF MATURE EXOGENOUS MIRNAS IN CLL CELLS WAS ESTABLISHED BY TRANSIENT TRANSFECTION. NDRG2 PROTEIN LEVELS IN CLL CELLS WERE DETECTED BY WESTERN BLOT. IN ADDITION, FLOW CYTOMETRY WAS CONDUCTED TO EXAMINE THE APOPTOSIS OF CLL CELLS. RESULTS: LOWER EXPRESSION OF NDRG2 WAS FOUND IN THE B-CELLS FROM 102 CLL PATIENTS COMPARED THE 40 NORMAL SUBJECTS (P < 0.001). PATIENTS WITH ADVANCED BINET STAGE (P = 0.001), HIGH LACTATE DEHYDROGENASE (LDH) LEVEL (P = 0.036), UN-MUTATED IMMUNOGLOBULIN HEAVY CHAIN VARIABLE REGION GENE (IGHV) (P = 0.004) AND THOSE WITH P53 ABERRATIONS (P < 0.001) HAD A MARKEDLY LOWER LEVELS OF NDRG2 MRNA. THIS DECREASE WAS ASSOCIATED WITH BRIEFER TIME-TO-TREATMENT (P = 0.001) AND POORER SURVIVAL (P < 0.001). HIGH EXPRESSION OF MIR-28-5P AND MIR-650 WAS ASSOCIATED WITH BINET B/C STAGE (P = 0.044) AND IGHV UN-MUTATED (P = 0.011), AS WELL AS BINET B/C STAGE (P = 0.013) AND P53 ABERRATIONS (P = 0.037), RESPECTIVELY. INHIBITION OF MIR-28-5P OR MIR-650 COULD INDUCE MORE APOPTOSIS IN CLL CELLS WITH GERMLINE TP53. CONCLUSIONS: NDRG2 MRNA LEVELS MIGHT BE A USEFUL PROGNOSTIC VARIABLE FOR PATIENTS OF CLL AND UP-REGULATING NDRG2 TRANSCRIPTION MAY BE A THERAPY APPROACH IN CLL WITHOUT P53 ABERRATIONS. 2018 7 3783 39 INTERFERON-GAMMA PROMOTER HYPOMETHYLATION AND INCREASED EXPRESSION IN CHRONIC PERIODONTITIS. AIM: THE GOAL OF THIS INVESTIGATION WAS TO DETERMINE WHETHER EPIGENETIC MODIFICATIONS IN THE IFNG PROMOTER ARE ASSOCIATED WITH AN INCREASE OF IFNG TRANSCRIPTION IN DIFFERENT STAGES OF PERIODONTAL DISEASES. MATERIALS AND METHODS: DNA WAS EXTRACTED FROM GINGIVAL BIOPSY SAMPLES COLLECTED FROM 47 TOTAL SITES FROM 47 DIFFERENT SUBJECTS: 23 PERIODONTALLY HEALTHY SITES, 12 EXPERIMENTALLY INDUCED GINGIVITIS SITES AND 12 CHRONIC PERIODONTITIS SITES. LEVELS OF DNA METHYLATION WITHIN THE IFNG PROMOTER CONTAINING SIX CPG DINUCLEOTIDES WERE DETERMINED USING PYROSEQUENCING TECHNOLOGY. INTERFERON GAMMA MRNA EXPRESSION WAS ANALYSED BY QUANTITATIVE POLYMERASE CHAIN REACTIONS USING ISOLATED RNA FROM PART OF THE BIOLOGICAL SAMPLES MENTIONED ABOVE. RESULTS: THE METHYLATION LEVEL OF ALL SIX ANALYSED CPG SITES WITHIN THE IFNG PROMOTER REGION IN THE PERIODONTITIS BIOPSIES 52% [INTERQUARTILE RANGE, IQR (43.8%, 63%)] WAS SIGNIFICANTLY LOWER THAN PERIODONTALLY HEALTHY SAMPLES 62% [IQR (51.3%, 74%)], P=0.007 AND GINGIVITIS BIOPSIES 63% [IQR (55%, 74%)], P=0.02. THE TRANSCRIPTIONAL LEVEL OF IFNG IN PERIODONTITIS BIOPSIES WAS 1.96-FOLD AND SIGNIFICANTLY HIGHER THAN TISSUES WITH PERIODONTAL HEALTH (P=0.04). ALTHOUGH THE MRNA LEVEL FROM EXPERIMENTAL GINGIVITIS SAMPLES EXHIBITED AN 8.5-FOLD INCREASE AS COMPARED WITH PERIODONTALLY HEALTHY SAMPLES, NO SIGNIFICANT METHYLATION DIFFERENCE WAS OBSERVED IN EXPERIMENTAL GINGIVITIS SAMPLE. CONCLUSIONS: A HYPOMETHYLATION PROFILE WITHIN IFNG PROMOTER REGION IS RELATED TO AN INCREASE OF IFNG TRANSCRIPTION PRESENT IN THE CHRONIC PERIODONTITIS BIOPSIES, WHILE SUCH AN INCREASE OF IFNG IN EXPERIMENTALLY INDUCED GINGIVITIS SEEMS INDEPENDENT OF PROMOTER METHYLATION ALTERATION. 2010 8 3460 34 HYPOMETHYLATION OF THE IL8 PROMOTER IN NASAL EPITHELIAL CELLS OF PATIENTS WITH CHRONIC RHINOSINUSITIS WITH NASAL POLYPS. BACKGROUND: IL-8 IS AN IMPORTANT CHEMOKINE IMPLICATED IN THE PATHOGENESIS OF CHRONIC RHINOSINUSITIS (CRS), BUT LITTLE IS KNOWN ABOUT EPIGENETIC REGULATION OF IL8 IN THE PATHOGENESIS OF CRS. OBJECTIVE: WE SOUGHT TO INVESTIGATE THE RELATIONSHIP BETWEEN THE DNA METHYLATION LEVEL IN THE IL8 PROXIMAL PROMOTER AND CRS IN HAN CHINESE SUBJECTS. METHODS: PATIENTS WITH CHRONIC RHINOSINUSITIS WITH NASAL POLYPS (CRSWNP; N = 187), PATIENTS WITH CHRONIC RHINOSINUSITIS WITHOUT NASAL POLYPS (CRSSNP; N = 89), AND CONTROL SUBJECTS (N = 57) WERE ENROLLED IN 2 INDEPENDENT COHORTS. PURIFIED HUMAN NASAL EPITHELIAL CELLS FROM EACH PARTICIPANT WERE ASSESSED FOR PERCENTAGE DNA METHYLATION OF CPG SITES IN THE IL8 PROXIMAL PROMOTER BY USING BISULFITE PYROSEQUENCING AND FOR FUNCTIONAL ASPECTS OF METHYLATION STATUS BY USING IN VITRO ASSAYS. RESULTS: DNA METHYLATION OF CPG SITES 1, 2, AND 3, RESPECTIVELY, IN THE IL8 PROXIMAL PROMOTER WAS SIGNIFICANTLY DECREASED IN HUMAN NASAL EPITHELIAL CELLS OF PATIENTS WITH CRSWNP COMPARED WITH THAT IN PATIENTS WITH CRSSNP (P < .001) AND CONTROL SUBJECTS (P < .001). PERCENTAGE OF DNA METHYLATION OF THE CPG3 SITE WAS CORRELATED NEGATIVELY WITH BOTH TISSUE EOSINOPHILIC CATIONIC PROTEIN (P < .01) AND MYELOPEROXIDASE (P < .05) LEVELS. IL-1BETA (P < .001) AND TNF-ALPHA (P < .01) SIGNIFICANTLY INCREASED IL8 EXPRESSION ACCOMPANIED BY A REDUCTION IN METHYLATION AT THE CPG3 SITE (P < .001). ELECTROPHORETIC MOBILITY SHIFT ASSAYS DEMONSTRATED THAT METHYLATION STATUS OF CPG3 CHANGED THE BINDING OF OCTAMER-BINDING TRANSCRIPTION FACTOR 1 AND NUCLEAR FACTOR KAPPAB. CONCLUSION: DECREASED DNA METHYLATION OF PARTICULARLY CPG SITES IN THE IL8 PROXIMAL PROMOTER MIGHT PLAY A ROLE IN THE PATHOGENESIS OF CRSWNP. 2019 9 3457 43 HYPOMETHYLATION OF INTERLEUKIN-6 (IL-6) GENE INCREASES THE RISK OF ESSENTIAL HYPERTENSION: A MATCHED CASE-CONTROL STUDY. ESSENTIAL HYPERTENSION (EH) IS A CHRONIC DISEASE WITH CLEAR EPIGENETIC COMPONENT. INFLAMMATION AND ENDOTHELIAL DYSFUNCTION HAVE A GREAT ROLE IN THE DEVELOPMENT OF PERSISTENT BLOOD PRESSURE ELEVATION. THE AIM OF THIS STUDY WAS TO EXPLORE AN ASSOCIATION BETWEEN EH AND DNA METHYLATION IN PRO-INFLAMMATION CYTOKINE GENE INTERLEUKIN-6 (IL-6) DURING THE INFLAMMATORY PROCESS. WE PERFORMED METHYLATION ANALYSIS OF PERIPHERAL BLOOD DNA USING BISULPHITE PYROSEQUENCING TECHNOLOGY BETWEEN 96 EH PATIENTS AND 96 AGE- AND GENDER-MATCHED HEALTHY CONTROLS. THE PRESENT RESULTS SHOWED THAT THREE CYTOSINE-PHOSPHATE-GUANINE (CPG) SITES OF IL-6 PROMOTER CPG ISLAND HAD DIFFERENT LOWER METHYLATION IN EH GROUP COMPARED WITH CONTROLS, BUT ONLY CPG2 (58.43+/-7.53 VERSUS 62.34+/-9.65, P=0.004) AND CPG3 (51.52+/-6.18 VERSUS 57.45+/-8.29, P<0.001) HAD STATISTICAL DIFFERENCE. LOGISTIC REGRESSION ANALYSIS FOUND CPG3 HYPOMETHYLATION WAS A RISK FACTOR OF EH (ODDS RATIO=1.111, ADJUSTED P=0.004). IN ADDITION, WE FOUND HYPERMETHYLATION OF CPG1 (64.84+/-7.06 VERSUS 61.84+/-8.61) AND CPG2 (62.04+/-7.40 VERSUS 59.30+/-9.57) IN MALE COMPARED WITH FEMALE. IN MALE, WE FOUND HYPOMETHYLATION OF CPG2 (60.41+/-7.74 VERSUS 64.28+/-6.36) AND CPG3 (53.70+/-8.62 VERSUS 57.78+/-7.87) OF SMOKER VERSUS NON-SMOKER AND HYPOMETHYLATION OF CPG2 (60.89+/-7.32 VERSUS 64.70+/-7.03) AND CPG3 (53.23+/-7.99 VERSUS 60.48+/-7.58) OF DRINKER VERSUS NON-DRINKER. FURTHERMORE, THE CPG2 AND CPG3 HAD A NEGATIVE CORRELATION WITH SYSTOLIC BLOOD PRESSURE AND DIASTOLIC BLOOD PRESSURE (P<0.05). RECEIVER OPERATING CHARACTERISTIC CURVE ANALYSIS SHOWED THAT CPG2 (AREA UNDER CURVE: 0.638, P=0.001) AND CPG3 (AREA UNDER CURVE: 0.704, P<0.001) HAD A DIAGNOSTIC VALUE TO PREDICT THE RISK OF EH. IN SUMMARY, OUR STUDY REVEALED HYPOMETHYLATION OF IL-6 WAS CORRELATED WITH THE RISK OF EH IN THE POPULATION ASSESSED AND WE FOUND THE DIFFERENCES OF IL-6 PROMOTER METHYLATION IN GENDER, SMOKING AND DRINKING. 2017 10 510 33 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 11 4245 33 METHYLATION STATUS OF DDIT3 GENE IN CHRONIC MYELOID LEUKEMIA. BACKGROUND: DNA-DAMAGE-INDUCIBLE TRANSCRIPT 3 (DDIT3), A CANDIDATE TUMOR SUPPRESSOR GENE (TSG), HAS BEEN FOUND INVOLVED IN THE REGULATION OF CELLULAR GROWTH AND DIFFERENTIATION. THE EPIGENETIC CHANGES OF TSGS ARE RECENTLY RECOGNIZED AS AN ABNORMAL MECHANISM CONTRIBUTING TO THE DEVELOPMENT OF CHRONIC MYELOID LEUKEMIA (CML). THE AIM OF THIS STUDY WAS TO INVESTIGATE THE METHYLATION STATUS OF DDIT3 GENE IN CML PATIENTS. METHODS: THE METHYLATION STATUS OF DDIT3 PROMOTER WAS DETECTED IN THE BONE MARROW MONONUCLEAR CELLS FROM 53 PATIENTS WITH CML USING METHYLATION-SPECIFIC PCR (MSP). THE EXPRESSION LEVELS OF DDIT3 AND BCR/ABL TRANSCRIPT WERE DETERMINED BY REAL-TIME QUANTITATIVE PCR (RQ-PCR). CLINICAL DATA OF THESE PATIENTS WERE COLLECTED AND ANALYZED. RESULTS: THE ABERRANT METHYLATION OF DDIT3 GENE PROMOTER WAS FOUND IN 35 OF 53 (66%) CML CASES. CORRELATION WAS NOT FOUND BETWEEN DDIT3 PROMOTER HYPERMETHYLATION AND THE AGE, SEX, HEMOGLOBIN CONCENTRATION, PLATELET COUNTS, CHROMOSOMAL ABNORMALITIES, BCR/ABL TRANSCRIPT, AND STAGING OF CML PATIENTS (P > 0.05), BUT FOUND BETWEEN DDIT3 PROMOTER HYPERMETHYLATION AND WBC COUNTS OF CML CASES (R = 0.781, P < 0.001). THE LEVEL OF DDIT3 TRANSCRIPT IN CML PATIENTS WAS SIGNIFICANTLY LOWER THAN THAT IN CONTROLS (MEDIAN 3.28 VS 19.69, P < 0.001), HOWEVER, THERE WAS NO DIFFERENCE IN THE LEVEL OF DDIT3 TRANSCRIPT BETWEEN METHYLATION-POSITIVE CML CASES (0.05-65.32, MEDIAN 2.13) AND METHYLATION- NEGATIVE CML CASES (0.12-126.04, MEDIAN 3.92) (P > 0.05). CONCLUSION: OUR RESULTS DEMONSTRATE THAT ABERRANT METHYLATION OF DDIT3 OCCURS IN CML FREQUENTLY. 2010 12 479 42 ARSENIC TRIOXIDE INHIBITS DNA METHYLTRANSFERASE AND RESTORES TMS1 GENE EXPRESSION IN K562 CELLS. BACKGROUND: GENE SILENCING ASSOCIATED WITH ABERRANT METHYLATION OF PROMOTER REGION CPG ISLANDS IS AN ACQUIRED EPIGENETIC ALTERATION THAT SERVES AS AN ALTERNATIVE TO GENETIC DEFECTS IN THE INACTIVATION OF TUMOR SUPPRESSOR GENES IN HUMAN CANCERS. THE DEMETHYLATING, DOSE-DEPENDENT EFFECT OF ARSENIC TRIOXIDE (AS2O3) ON SEVERAL TUMOR-RELATED GENES HAS ALREADY BEEN POSTULATED. HOWEVER, WHETHER SUCH A DEMETHYLATING EFFECT ALSO APPLIES TO THE TMS1 GENE IN CHRONIC MYELOID LEUKEMIA CELL LINE K562 CELLS HAS NOT BEEN STUDIED SO FAR. THE AIM OF THE PRESENT STUDY WAS TO DETECT THE METHYLATION STATUS OF THE TMS1 GENE IN K562 CELLS AND THE DEMETHYLATION EFFECT OF AS2O3 ON TMS1 AS WELL AS TMS1 APOPTOSIS-ASSOCIATED PROTEIN BCL-2/BAX AND DNA METHYLTRANSFERASE (DNMT) EXPRESSION. METHODS: TMS1 MRNA EXPRESSION IN K562 CELLS AND NORMAL BONE MARROW WAS DETERMINED BY REVERSE TRANSCRIPTION (RT) POLYMERASE CHAIN REACTION (PCR), AND THE DNA METHYLATION STATUS OF THE TMS1 PROMOTER IN K562 CELLS TREATED WITH DIFFERENT CONCENTRATIONS OF AS2O3 FOR 48 H WAS DETERMINED BY METHYLATION-SPECIFIC PCR. RT-PCR AND WESTERN BLOT WERE USED TO DETECT TMS1 AND DNMT EXPRESSION. WE ALSO ASSESSED TMS1-ASSOCIATED APOPTOSIS PROTEIN BCL-2/BAX EXPRESSION BY WESTERN BLOT AND APOPTOSIS RATES BY FLOW CYTOMETRY USING ANNEXIN V/PROPIDIUM IODIDE DOUBLE STAINING. RESULTS: IN K562 CELLS, TMS1 WAS COMPLETELY METHYLATED AND BOTH TMS1 MRNA AND PROTEIN SHOWED A LOW EXPRESSION, BUT 2 MUMOL/L AS2O3 COULD SIGNIFICANTLY RESTORE THE EXPRESSION OF THE TMS1 GENE BOTH AT MRNA AND PROTEIN LEVEL (P < 0.01) BY FULLY REVERSING DNA METHYLATION. AS2O3 DECREASED MRNA AND PROTEIN EXPRESSION OF DNMT1 (P < 0.05) IN A DOSE-DEPENDENT MANNER. FLOW CYTOMETRY SHOWED THAT IN THE EXPERIMENTAL GROUP (2 MUMOL/L AS2O3), CELL APOPTOSIS WAS SIGNIFICANTLY INCREASED COMPARED WITH THE CONTROL GROUP (NO AS2O3; P < 0.05). IN THE EXPERIMENTAL GROUP, WESTERN BLOT SHOWED THAT THE EXPRESSION OF THE ANTI-APOPTOTIC PROTEIN BCL-2 WAS SIGNIFICANTLY DECREASED; HOWEVER, THE PROAPOPTOTIC PROTEIN BAX WAS MARKEDLY INCREASED AND THE BCL-2/BAX RATIO WAS MARKEDLY REDUCED (P < 0.01). CONCLUSIONS: AS2O3 COULD RESTORE THE EXPRESSION OF TMS1 BY INHIBITING DNMT TO REVERSE THE HYPERMETHYLATION AND INDUCED APOPTOSIS OF K562 CELLS BY DOWNREGULATION OF BCL-2/BAX EXPRESSION. 2015 13 3044 39 GENOME-WIDE ANALYSIS OF 5-HMC IN THE PERIPHERAL BLOOD OF SYSTEMIC LUPUS ERYTHEMATOSUS PATIENTS USING AN HMEDIP-CHIP. SYSTEMIC LUPUS ERYTHEMATOSUS (SLE) IS A CHRONIC, POTENTIALLY FATAL SYSTEMIC AUTOIMMUNE DISEASE CHARACTERIZED BY THE PRODUCTION OF AUTOANTIBODIES AGAINST A WIDE RANGE OF SELF-ANTIGENS. TO INVESTIGATE THE ROLE OF THE 5-HMC DNA MODIFICATION WITH REGARD TO THE ONSET OF SLE, WE COMPARED THE LEVELS 5-HMC BETWEEN SLE PATIENTS AND NORMAL CONTROLS. WHOLE BLOOD WAS OBTAINED FROM PATIENTS, AND GENOMIC DNA WAS EXTRACTED. USING THE HMEDIP-CHIP ANALYSIS AND VALIDATION BY QUANTITATIVE RT-PCR (RT-QPCR), WE IDENTIFIED THE DIFFERENTIALLY HYDROXYMETHYLATED REGIONS THAT ARE ASSOCIATED WITH SLE. THERE WERE 1,701 GENES WITH SIGNIFICANTLY DIFFERENT 5-HMC LEVELS AT THE PROMOTER REGION IN THE SLE PATIENTS COMPARED WITH THE NORMAL CONTROLS. THE CPG ISLANDS OF 3,826 GENES SHOWED SIGNIFICANTLY DIFFERENT 5-HMC LEVELS IN THE SLE PATIENTS COMPARED WITH THE NORMAL CONTROLS. OUT OF THE DIFFERENTIALLY HYDROXYMETHYLATED GENES, THREE WERE SELECTED FOR VALIDATION, INCLUDING TREX1, CDKN1A AND CDKN1B. THE HYDROXYMETHYLATION LEVELS OF THE THREE GENES WERE CONFIRMED BY RT-QPCR. THE RESULTS SUGGESTED THAT THERE WERE SIGNIFICANT ALTERATIONS OF 5-HMC IN SLE PATIENTS. THUS, THESE DIFFERENTIALLY HYDROXYMETHYLATED GENES MAY CONTRIBUTE TO THE PATHOGENESIS OF SLE. THESE FINDINGS SHOW THE SIGNIFICANCE OF 5-HMC AS A POTENTIAL BIOMARKER OR PROMISING TARGET FOR EPIGENETIC-BASED SLE THERAPIES. 2015 14 4350 34 MIR-181A-5P IS A POTENTIAL CANDIDATE EPIGENETIC BIOMARKER IN MULTIPLE SCLEROSIS. MULTIPLE SCLEROSIS (MS) IS A CHRONIC INFLAMMATORY DISEASE OF THE CENTRAL NERVOUS SYSTEM (CNS) CHARACTERIZED BY DEMYELINATION AND AXONAL DEGENERATION. ABNORMAL EXPRESSION OF MICRORNAS (MIRNAS) PLAYS AN IMPORTANT ROLE IN MS PATHOLOGY. IN THIS COHORT STUDY, DIFFERENTIAL EXPRESSION OF THE FOUR MIRNAS (HSA-MIR-155-5P, HSA-MIR-9-5P, HSA-MIR-181A-5P, AND HSA-MIR-125B-5P) WAS INVESTIGATED IN 69 INDIVIDUALS, INCLUDING 39 MS PATIENTS (RELAPSING-REMITTING MS (RRMS), N = 27; SECONDARY PROGRESSIVE MS (SPMS), N = 12) AND 30 HEALTHY CONTROLS. IN SILICO ANALYSES REVEALED POSSIBLE GENES AND PATHWAYS SPECIFIC TO MIRNAS. PERIPHERAL BLOOD MIRNA EXPRESSIONS WERE DETECTED BY QUANTITATIVE REAL-TIME PCR (QPCR). HSA-MIR-181A-5P WAS DOWNREGULATED AND ASSOCIATED WITH INCREASED MS RISK (P = 0.012). THE OTHER THREE MIRNAS WERE UPREGULATED AND NOT ASSOCIATED WITH MS (P < 0.05). THE AREA UNDER THE CURVE (AUC) IS 0.779. IN SILICO ANALYSES SHOWED THAT HSA-MIR-181A-5P MAY PARTICIPATE IN MS PATHOLOGY BY TARGETING MAP2K1, CREB1, ATXN1, AND ATXN3 GENES IN INFLAMMATION AND NEURODEGENERATION PATHWAYS. THE CIRCULATORY HSA-MIR-181A-5P CAN REGULATE TARGET GENES, REVERSING THE MECHANISMS INVOLVED IN MS PATHOLOGIES SUCH AS PROTEIN UPTAKE AND PROCESSING, CELL PROLIFERATION AND SURVIVAL, INFLAMMATION, AND NEURODEGENERATION. THUS, THIS MIRNA COULD BE USED AS AN EPIGENOMIC-GUIDED DIAGNOSTIC TOOL AND FOR THERAPEUTIC PURPOSE. 2022 15 1029 34 CIRCULATING PLASMA MICRORNA IN PATIENTS WITH ACTIVE ACROMEGALY. CONTEXT: EXCESSIVE PRODUCTION OF GROWTH HORMONE CAUSES MARKED MULTIORGAN CHANGES IN PATIENTS WITH ACROMEGALY, WHICH MAY INVOLVE EPIGENETIC MECHANISMS. OBJECTIVE: TO EVALUATE DIFFERENCES IN CIRCULATING MICRORNAS (MIRNAS) ASSOCIATED WITH CHRONIC GROWTH HORMONE OVERPRODUCTION IN ADULTS. DESIGN AND SETTING: A CROSS-SECTIONAL CASE-CONTROL STUDY WAS CONDUCTED AT A TERTIARY MEDICAL CENTER. PARTICIPANTS: WE ENROLLED 12 CONSECUTIVE PATIENTS WITH ACROMEGALY ALONG WITH 12 AGE- AND SEX-MATCHED CONTROLS IN THE DISCOVERY PHASE OF THE STUDY AND THEN EXTENDED THIS COHORT TO 47 PATIENTS WITH ACROMEGALY AND 28 HEALTHY CONTROLS FOR THE VALIDATION STUDY. MAIN OUTCOME MEASURES: PLASMA MIRNAS WERE QUANTIFIED BY NEXT-GENERATION SEQUENCING (NGS) IN THE DISCOVERY PHASE. LEVELS OF SELECTED MIRNAS WERE VALIDATED ON EXTENDED COHORTS USING REVERSE TRANSCRIPTION QUANTITATIVE POLYMERASE CHAIN REACTION (RT-QPCR), COMPARED BETWEEN GROUPS, AND CORRELATED WITH CLINICAL PARAMETERS. RESULTS: BASED ON NGS DATA, WE SELECTED 3 PLASMA MIRNAS DOWNREGULATED IN PATIENTS WITH ACROMEGALY COMPARED TO HEALTHY CONTROLS: MIR-4446-3P -1.317 (P = 0.001), MIR-215-5P -3.040 (P = 0.005), AND MIR-342-5P -1.875 (P = 0.013) WITHOUT MULTIPLICITY CORRECTION FOR ALL 3 MIRNAS. THESE RESULTS WERE CONFIRMED BY RT-QPCR IN THE VALIDATION PHASE FOR 2 MIRNAS OUT OF 3: MIR-4446-3P (P < 0.001, PADJUSTED < 0.001), AREA UNDER THE RECEIVER-OPERATOR CURVE (AUC) 0.862 (95% CI 0.723-0.936; P < 0.001) AND MIR-215-5P (P < 0.001, PADJUSTED < 0.001), AUC 0.829 (95% CI 0.698-0.907; P < 0.001) TO DIFFERENTIATE PATIENTS WITH ACROMEGALY FROM HEALTHY CONTROLS. CONCLUSIONS: IN A 2-PHASE EXPERIMENT USING 2 DIFFERENT TECHNIQUES WE FOUND AND VALIDATED THE DOWNREGULATION OF PLASMA MIR-4446-3P AND MIR-215-5P IN PATIENTS WITH ACROMEGALY COMPARED TO HEALTHY SUBJECTS, WHICH MAKES THEM PROMISING BIOMARKERS FOR FURTHER RESEARCH. 2022 16 6589 35 TUMOR NECROSIS FACTOR-ALPHA GENE PROMOTER METHYLATION IN JAPANESE ADULTS WITH CHRONIC PERIODONTITIS AND RHEUMATOID ARTHRITIS. BACKGROUND AND OBJECTIVE: OVER-EXPRESSION OF TUMOR NECROSIS FACTOR-ALPHA (TNF-ALPHA) PLAYS A PATHOLOGICAL ROLE IN CHRONIC PERIODONTITIS (CP) AND RHEUMATOID ARTHRITIS (RA), WHICH MIGHT BE REGULATED BY THE EPIGENETIC MECHANISM. THE AIM OF THE PRESENT STUDY WAS TO EVALUATE WHETHER THERE IS A UNIQUE METHYLATION PROFILE OF THE TNF-ALPHA GENE PROMOTER IN BLOOD CELLS OF INDIVIDUALS WITH CP AND RA. MATERIAL AND METHODS: THE STUDY PARTICIPANTS CONSISTED OF 30 JAPANESE ADULTS WITH RA (RA GROUP), 30 RACE-MATCHED ADULTS WITH CP ONLY (CP GROUP) AND 30 RACE-MATCHED HEALTHY CONTROLS (H GROUP). GENOMIC DNA ISOLATED FROM PERIPHERAL BLOOD WAS MODIFIED BY SODIUM BISULFITE AND ANALYZED, BY DIRECT SEQUENCING, TO INVESTIGATE DNA METHYLATION OF THE TNF-ALPHA GENE PROMOTER REGION. THE LEVEL OF TNF-ALPHA PRODUCED IN MONONUCLEAR CELLS STIMULATED WITH PORPHYROMONAS GINGIVALIS LIPOPOLYSACCHARIDE WAS DETERMINED USING ELISA. RESULTS: TWELVE CYTOSINE-GUANINE DINUCLEOTIDE (CPG) MOTIFS WERE IDENTIFIED IN THE TNF-ALPHA PROMOTER FRAGMENT FROM -343 TO +57 BP. THE CP GROUP SHOWED A SIGNIFICANTLY HIGHER METHYLATION RATE AND FREQUENCY AT -72 BP THAN THE H GROUP (P < 0.01). THE RA GROUP EXHIBITED SIGNIFICANTLY HIGHER METHYLATION RATES AT SEVEN CPG MOTIFS (-302, -163, -119, -72, -49, -38 AND +10 BP), AND SIGNIFICANTLY HIGHER METHYLATION FREQUENCIES AT SIX CPG MOTIFS (-163, -119, -72, -49, -38 AND +10 BP), THAN THE H GROUP (P < 0.01 FOR ALL COMPARISONS). THE LEVELS OF TNF-ALPHA PRODUCED WERE SIGNIFICANTLY DIFFERENT BETWEEN INDIVIDUALS WITH AND WITHOUT METHYLATION AT -163 BP (P = 0.03). CONCLUSION: THESE RESULTS SUGGEST THAT THE HYPERMETHYLATED STATUS OF CPG MOTIFS IN THE TNF-ALPHA GENE PROMOTER IN BLOOD CELLS MAY BE UNIQUE TO JAPANESE ADULTS WITH CP AND RA. 2016 17 4021 29 LOWERED DNA METHYLTRANSFERASE (DNMT-3B) MRNA EXPRESSION IS ASSOCIATED WITH GENOMIC DNA HYPERMETHYLATION IN PATIENTS WITH CHRONIC ALCOHOLISM. DNA METHYLTRANSFERASES (DNMTS) ARE INVOLVED WITHIN THE EPIGENETIC CONTROL OF DNA METHYLATION PROCESSES. RECENTLY, IT HAS BEEN SHOWN THAT THE GENOMIC DNA METHYLATION IN PATIENTS WITH ALCOHOLISM IS INCREASED. IN THE PRESENT CONTROLLED STUDY WE OBSERVED A SIGNIFICANT DECREASE OF MRNA EXPRESSION OF DNMT-3A AND DNMT-3B WHEN COMPARING ALCOHOLIC PATIENTS (N = 59) WITH HEALTHY CONTROLS (N = 66): DNMT-3A (T = -2.38, P = 0.019), DNMT-3B (T = -2.65, P = 0.008). NO SIGNIFICANT DIFFERENCES WERE SEEN FOR DNMT-1 AND MBD-2 (METHYL-CPG-BINDING-DOMAIN PROTEIN 2) EXPRESSION. ADDITIONALLY, WE OBSERVED A SIGNIFICANT NEGATIVE CORRELATION BETWEEN DNMT-3B EXPRESSION AND THE BLOOD ALCOHOL CONCENTRATION (R = -0.45, P = 0.003) WHICH MIGHT EXPLAIN THE DECREASE OF DNMT-3B MRNA EXPRESSION IN ALCOHOLIC PATIENTS. USING A MULTIVARIATE MODEL WE OBSERVED THAT THE INCREASE (10%) OF GENOMIC DNA METHYLATION IN PATIENTS WITH ALCOHOLISM WAS SIGNIFICANTLY ASSOCIATED WITH THEIR LOWERED DNMT-3B MRNA EXPRESSION (MULTIPLE LINEAR REGRESSION, P = 0.014). SINCE METHYLATION OF DNA IS AN IMPORTANT EPIGENETIC FACTOR IN REGULATION OF GENE EXPRESSION THESE FINDINGS MAY HAVE IMPORTANT IMPLICATIONS FOR A POSSIBLE SUBSEQUENT DERANGEMENT OF EPIGENETIC CONTROL IN THESE PATIENTS. 2006 18 1297 44 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 19 5086 36 PILOT STUDY OF DNA METHYLATION IN THE PATHOGENESIS OF CHRONIC RHINOSINUSITIS WITH NASAL POLYPS. BACKGROUND: DNA METHYLATION HAS BEEN IMPLICATED IN THE PATHOGENESIS OF ALLERGY AND ATOPY. THIS STUDY AIMED TO IDENTIFY WHETHER DNA METHYLATION ALSO PLAYS AN IMPORTANT ROLE IN THE PATHOGENESIS OF NASAL POLYPS (NP). METHODOLOGY: NP TISSUES WERE OBTAINED FROM 32 PATIENTS WITH CHRONIC RHINOSINUSITIS WITH BILATERAL NP. BIOPSIES OF INFERIOR TURBINATE MUCOSA (ITM) WERE TAKEN FROM 18 PATIENTS WHO UNDERWENT RHINOSEPTOPLASTY (CONTROL GROUP). THE METHYLATED GENES, WHICH WERE DETECTED BY DNA METHYLATION MICROARRAY, WERE VALIDATED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION, BISULPHITE SEQUENCING, REAL-TIME POLYMERASE CHAIN REACTION AND IMMUNOHISTOCHEMISTRY. RESULTS: DNA METHYLATION MICROARRAY IDENTIFIED 8,008 CPG ISLANDS IN 2,848 GENES. ONE HUNDRED AND NINETY-EIGHT GENES WERE FOUND TO HAVE A METHYLATED SIGNAL IN THE PROMOTER REGION IN NP SAMPLES COMPARED WITH ITM SAMPLES. THE FOUR TOP GENES THAT CHANGED, COL18A1, EP300, GNAS AND SMURF1, WERE SELECTED FOR FURTHER STUDY. THE METHYLATION FREQUENCY OF COL18A1 WAS SIGNIFICANTLY HIGHER IN NP SAMPLES THAN IN ITM SAMPLES. CONCLUSIONS: DNA METHYLATION MIGHT PLAY AN IMPORTANT ROLE IN THE PATHOGENESIS OF NP. PROMOTER METHYLATION OF COL18A1 WAS FOUND TO BE SIGNIFICANTLY INCREASED IN NP TISSUES, FURTHER STUDIES ARE NECESSARY TO CONFIRM THE SIGNIFICANCE OF THESE EPIGENETIC FACTORS IN THE MECHANISMS UNDERLYING THE DEVELOPMENT OR PERSISTENCE OF NP. 2015 20 6170 28 THE HDAC INHIBITOR SAHA IMPROVES DEPRESSIVE-LIKE BEHAVIOR OF CRTC1-DEFICIENT MICE: POSSIBLE RELEVANCE FOR TREATMENT-RESISTANT DEPRESSION. MAJOR DEPRESSION IS A HIGHLY COMPLEX DISABLING PSYCHIATRIC DISORDER AFFECTING MILLIONS OF PEOPLE WORLDWIDE. DESPITE THE AVAILABILITY OF SEVERAL CLASSES OF ANTIDEPRESSANTS, A SUBSTANTIAL PERCENTAGE OF PATIENTS ARE UNRESPONSIVE TO THESE MEDICATIONS. A BETTER UNDERSTANDING OF THE NEUROBIOLOGY OF DEPRESSION AND THE MECHANISMS UNDERLYING ANTIDEPRESSANT RESPONSE IS THUS CRITICALLY NEEDED. WE PREVIOUSLY REPORTED THAT MICE LACKING CREB-REGULATED TRANSCRIPTION COACTIVATOR 1 (CRTC1) EXHIBIT A DEPRESSIVE-LIKE PHENOTYPE AND A BLUNTED ANTIDEPRESSANT RESPONSE TO THE SELECTIVE SEROTONIN REUPTAKE INHIBITOR FLUOXETINE. IN THIS STUDY, WE SIMILARLY SHOW THAT CRTC1(-/-) MICE ARE RESISTANT TO THE ANTIDEPRESSANT EFFECT OF CHRONIC DESIPRAMINE IN A BEHAVIORAL DESPAIR PARADIGM. SUPPORTING THE BLUNTED RESPONSE TO THIS TRICYCLIC ANTIDEPRESSANT, WE FOUND THAT DESIPRAMINE DOES NOT SIGNIFICANTLY INCREASE THE EXPRESSION OF BDNF AND NR4A1-3 IN THE HIPPOCAMPUS AND PREFRONTAL CORTEX OF CRTC1(-/-) MICE. EPIGENETIC REGULATION OF NEUROPLASTICITY GENE EXPRESSION HAS BEEN ASSOCIATED WITH DEPRESSION AND ANTIDEPRESSANT RESPONSE, AND HISTONE DEACETYLASE (HDAC) INHIBITORS HAVE BEEN SHOWN TO HAVE ANTIDEPRESSANT-LIKE PROPERTIES. HERE, WE SHOW THAT UNLIKE CONVENTIONAL ANTIDEPRESSANTS, CHRONIC SYSTEMIC ADMINISTRATION OF THE HDAC INHIBITOR SAHA PARTIALLY RESCUES THE DEPRESSIVE-LIKE BEHAVIOR OF CRTC1(-/-) MICE. THIS BEHAVIORAL EFFECT IS ACCOMPANIED BY AN INCREASED EXPRESSION OF BDNF, BUT NOT NR4A1-3, IN THE PREFRONTAL CORTEX OF THESE MICE, SUGGESTING THAT THIS EPIGENETIC INTERVENTION RESTORES THE EXPRESSION OF A SUBSET OF GENES BY ACTING DOWNSTREAM OF CRTC1. THESE FINDINGS SUGGEST THAT CRTC1 ALTERATIONS MAY BE ASSOCIATED WITH TREATMENT-RESISTANT DEPRESSION, AND SUPPORT THE INTERESTING POSSIBILITY THAT TARGETING HDACS MAY BE A USEFUL THERAPEUTIC STRATEGY IN ANTIDEPRESSANT DEVELOPMENT. 2016