1  908 135 CHRONIC EXPOSURE TO ENVIRONMENTALLY RELEVANT CONCENTRATION OF FLUORIDE IMPAIRS OSTEOBLAST'S COLLAGEN SYNTHESIS AND MATRIX MINERALIZATION: INVOLVEMENT OF EPIGENETIC REGULATION IN SKELETAL FLUOROSIS. GLOBALLY, 200 MILLION PEOPLE ARE SUFFERING FROM TOXIC MANIFESTATIONS OF FLUORIDE(F), DENTAL AND SKELETAL FLUOROSIS; UNFORTUNATELY, THERE IS NO TREATMENT. TO UNRAVEL THE PATHOGENESIS OF SKELETAL FLUOROSIS, WE ESTABLISHED FLUOROSIS MICE BY TREATING ENVIRONMENTALLY RELEVANT CONCENTRATION OF F (15 PPM NAF) THROUGH DRINKING WATER FOR 4 MONTHS. AS IN SKELETAL FLUOROSIS, LOCOMOTOR DISABILITY, CRIPPLING DEFORMITIES OCCUR AND THUS, OUR HYPOTHESIS WAS F MIGHT ADVERSELY AFFECTS COLLAGEN WHICH GIVES THE BONE TENSILE STRENGTH. THIS WORK INEVITABLY HAD TO BE CARRIED OUT ON OSTEOBLAST CELLS, RESPONSIBLE FOR SYNTHESIS, DEPOSITION, AND MINERALIZATION OF BONE MATRIX. ISOLATED OSTEOBLAST CELLS WERE CONFIRMED BY ALP ACTIVITY AND MINERALIZED NODULES FORMATION. EXPRESSION OF COLLAGEN COL1A1, COL1A2, COL1A1 WAS SIGNIFICANTLY REDUCED IN TREATED MICE. FURTHER, A STUDY REVEALED THE INVOLVEMENT OF EPIGENETIC REGULATION BY PROMOTER HYPERMETHYLATION OF COL1A1; EXPRESSIONAL ALTERATIONS OF TRANSCRIPTION FACTORS, CALCIUM CHANNELS AND OTHER GENES E.G., CBFA-1, TGF-BETA1, BMP1, SP1, SP7, NF-(K)B P65, BMP-2, BGLAP, GPRC6A AND CAV(1.2) ARE ASSOCIATED WITH IMPAIRMENT OF COLLAGEN SYNTHESIS, DEPOSITION AND DECREASED MINERALIZATION THUS, ENFEEBLING BONE HEALTH. THIS STUDY INDICATES THE POSSIBLE ASSOCIATION OF EPIGENETIC REGULATION IN SKELETAL FLUOROSIS. HOWEVER, NO ASSOCIATION WAS FOUND BETWEEN POLYMORPHISMS IN THE COL1A1 (RSAI, HINDIII) AND COL1A2 (RSAI, HINDIII) GENES WITH FLUOROSIS IN MICE.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
2 5555  43 ROLE OF FLUORIDE INDUCED EPIGENETIC ALTERATIONS IN THE DEVELOPMENT OF SKELETAL FLUOROSIS. FLUORIDE IS AN ESSENTIAL TRACE ELEMENT REQUIRED FOR PROPER BONE AND TOOTH DEVELOPMENT. SYSTEMIC HIGH EXPOSURE TO FLUORIDE THROUGH ENVIRONMENTAL EXPOSURE (DRINKING WATER AND FOOD) MAY RESULT IN TOXICITY CAUSING A DISORDER CALLED FLUOROSIS. IN THE PRESENT STUDY, WE INVESTIGATED THE ALTERATION IN DNA METHYLATION PROFILE WITH CHRONIC EXPOSURE (30 DAYS) TO FLUORIDE (8 MG/L) AND ITS RELEVANCE IN THE DEVELOPMENT OF FLUOROSIS. WHOLE GENOME BISULFITE SEQUENCING (WGBS) WAS CARRIED OUT IN HUMAN OSTEOSARCOMA CELLS (HOS) EXPOSED TO FLUORIDE. WHOLE GENOME BISULFITE SEQUENCING (WGBS) AND FUNCTIONAL ANNOTATION OF DIFFERENTIALLY METHYLATED GENES INDICATE ALTERATIONS IN METHYLATION STATUS OF GENES INVOLVED IN BIOLOGICAL PROCESSES ASSOCIATED WITH BONE DEVELOPMENT PATHWAYS. COMBINED ANALYSIS OF PROMOTER DNA HYPER METHYLATION, STRING: FUNCTIONAL PROTEIN ASSOCIATION NETWORKS AND GENE EXPRESSION ANALYSIS REVEALED EPIGENETIC ALTERATIONS IN BMP1, METAP2, MMP11 AND BACH1 GENES, WHICH PLAYS A ROLE IN THE EXTRACELLULAR MATRIX DISASSEMBLY, COLLAGEN CATABOLIC/ORGANIZATION PROCESS, SKELETAL MORPHOGENESIS/DEVELOPMENT, OSSIFICATION AND OSTEOBLAST DEVELOPMENT. THE PRESENT STUDY SHOWS THAT FLUORIDE CAUSES PROMOTER DNA HYPERMETHYLATION IN BMP1, METAP2, MMP11 AND BACH1 GENES WITH SUBSEQUENT DOWN-REGULATION IN THEIR EXPRESSION LEVEL (RNA LEVEL). THE RESULTS IMPLIES THAT FLUORIDE INDUCED DNA HYPERMETHYLATION OF THESE GENES MAY HAMPER EXTRACELLULAR MATRIX DEPOSITION, CARTILAGE FORMATION, ANGIOGENESIS, VASCULAR SYSTEM DEVELOPMENT AND POROSITY OF BONE, THUS PROMOTE SKELETAL FLUOROSIS.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
3 5556  40 ROLE OF FLUORIDE INDUCED HISTONE TRIMETHYLATION IN DEVELOPMENT OF SKELETAL FLUOROSIS. CHRONIC EXPOSURE TO FLUORIDE HAS BEEN ASSOCIATED WITH THE DEVELOPMENT OF SKELETAL FLUOROSIS. LIMITED REPORTS ARE AVAILABLE ON FLUORIDE INDUCED HISTONE MODIFICATION. HOWEVER, THE ROLE OF HISTONE MODIFICATION IN THE PATHOGENESIS OF SKELETAL FLUOROSIS IS NOT INVESTIGATED. IN THE PRESENT STUDY, WE HAVE INVESTIGATED THE ROLE OF FLUORIDE INDUCED HISTONE MODIFICATION ON FLUOROSIS DEVELOPMENT USING HUMAN OSTEOSARCOMA (HOS) CELL LINE. THE EXPRESSION OF HISTONE METHYLTRANSFERASES (EHMT1 AND EHZ2) AND LEVEL OF GLOBAL HISTONE TRIMETHYLATION (H3K9 AND H3K27) HAVE BEEN ASSESSED AND OBSERVED TO BE INCREASED SIGNIFICANTLY AFTER FLUORIDE EXPOSURE (8 MG/L). EPITECT CHROMATIN IMMUNOPRECIPITATION (CHIP) QPCR ARRAY (HUMAN TGFBETA/BMP SIGNALING PATHWAY) WAS PERFORMED TO ASSESS THE H3K9 TRIMETHYLATION AT PROMOTER REGIONS OF PATHWAY-SPECIFIC GENES. H3K9 CHIP PCR ARRAY ANALYSIS IDENTIFIED HYPER H3K9 TRIMETHYLATION IN PROMOTER REGIONS OF TGFBR2 AND SMAD3. QPCR AND STRING ANALYSIS WAS CARRIED OUT TO DETERMINE THE REPRESSIVE EPIGENETIC EFFECT OF H3K9 TRIMETHYLATION ON EXPRESSION PATTERN AND FUNCTIONAL ASSOCIATION OF IDENTIFIED GENES. IDENTIFIED GENES (TGFBR2 AND SMAD3) SHOWED DOWN-REGULATION WHICH CONFIRMS THE REPRESSIVE EPIGENETIC EFFECT OF PROMOTER H3K9 HYPER TRIMETHYLATION. EXPRESSION OF TWO OTHER VITAL GENES COL1A1 AND MMP13 INVOLVED IN TGFBR2-SMAD SIGNALING PATHWAY WAS ALSO FOUND TO BE DOWN-REGULATED WITH A DECREASE IN EXPRESSION OF TGFBR2 AND SMAD3. STRING ANALYSIS REVEALED FUNCTIONAL ASSOCIATION AND INVOLVEMENT OF IDENTIFIED GENES TGFBR2, SMAD3, COL1A1 AND MMP13 IN THE COLLAGEN AND CARTILAGE DEVELOPMENT/MORPHOGENESIS, CONNECTIVE TISSUE FORMATION, BIO-MINERAL TISSUE DEVELOPMENT, ENDOCHONDRAL BONE FORMATION, BONE AND SKELETAL MORPHOGENESIS. IN CONCLUSION, PRESENT INVESTIGATION IS A FIRST ATTEMPT TO LINK FLUORIDE INDUCED HYPER H3K9 TRI-METHYLATION MEDIATED REPRESSION OF TGFBR2 AND SMAD3 WITH THE DEVELOPMENT OF SKELETAL FLUOROSIS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
4 3468  37 HYPOXIA-INDUCED DNA HYPERMETHYLATION IN HUMAN PULMONARY FIBROBLASTS IS ASSOCIATED WITH THY-1 PROMOTER METHYLATION AND THE DEVELOPMENT OF A PRO-FIBROTIC PHENOTYPE. BACKGROUND: PULMONARY FIBROSIS IS A DEBILITATING AND LETHAL DISEASE WITH NO EFFECTIVE TREATMENT OPTIONS. UNDERSTANDING THE PATHOLOGICAL PROCESSES AT PLAY WILL DIRECT THE APPLICATION OF NOVEL THERAPEUTIC AVENUES. HYPOXIA HAS BEEN IMPLICATED IN THE PATHOGENESIS OF PULMONARY FIBROSIS YET THE PRECISE MECHANISM BY WHICH IT CONTRIBUTES TO DISEASE PROGRESSION REMAINS TO BE FULLY ELUCIDATED. IT HAS BEEN SHOWN THAT CHRONIC HYPOXIA CAN ALTER DNA METHYLATION PATTERNS IN TUMOUR-DERIVED CELL LINES. THIS EPIGENETIC ALTERATION CAN INDUCE CHANGES IN CELLULAR PHENOTYPE WITH PROMOTER METHYLATION BEING ASSOCIATED WITH GENE SILENCING. OF PARTICULAR RELEVANCE TO IDIOPATHIC PULMONARY FIBROSIS (IPF) IS THE OBSERVATION THAT THY-1 PROMOTER METHYLATION IS ASSOCIATED WITH A MYOFIBROBLAST PHENOTYPE WHERE LOSS OF THY-1 OCCURS ALONGSIDE INCREASED ALPHA SMOOTH MUSCLE ACTIN (ALPHA-SMA) EXPRESSION. THE INITIAL AIM OF THIS STUDY WAS TO DETERMINE WHETHER HYPOXIA REGULATES DNA METHYLATION IN NORMAL HUMAN LUNG FIBROBLASTS (CCD19LU). AS IT HAS BEEN REPORTED THAT HYPOXIA SUPPRESSES THY-1 EXPRESSION DURING LUNG DEVELOPMENT WE ALSO STUDIED THE EFFECT OF HYPOXIA ON THY-1 PROMOTER METHYLATION AND GENE EXPRESSION. METHODS: CCD19LU WERE GROWN FOR UP TO 8 DAYS IN HYPOXIA AND ASSESSED FOR GLOBAL CHANGES IN DNA METHYLATION USING FLOW CYTOMETRY. REAL-TIME PCR WAS USED TO QUANTIFY EXPRESSION OF THY-1, ALPHA-SMA, COLLAGEN I AND III. GENOMIC DNA WAS BISULPHITE TREATED AND METHYLATION SPECIFIC PCR (MSPCR) WAS USED TO EXAMINE THE METHYLATION STATUS OF THE THY-1 PROMOTER. RESULTS: SIGNIFICANT GLOBAL HYPERMETHYLATION WAS DETECTED IN HYPOXIC FIBROBLASTS RELATIVE TO NORMOXIC CONTROLS AND WAS ACCOMPANIED BY INCREASED EXPRESSION OF MYOFIBROBLAST MARKERS. THY-1 MRNA EXPRESSION WAS SUPPRESSED IN HYPOXIC CELLS, WHICH WAS RESTORED WITH THE DEMETHYLATING AGENT 5-AZA-2'-DEOXYCYTIDINE. MSPCR REVEALED THAT THY-1 BECAME METHYLATED FOLLOWING FIBROBLAST EXPOSURE TO 1% O2. CONCLUSION: THESE DATA SUGGEST THAT GLOBAL AND GENE-SPECIFIC CHANGES IN DNA METHYLATION MAY PLAY AN IMPORTANT ROLE IN FIBROBLAST FUNCTION IN HYPOXIA.	2012	
                                                                                                                                                                                                                                                                       
5 1965  37 EPIGENETIC ALTERATION OF MITOCHONDRIAL BIOGENESIS REGULATORY GENES IN ARSENIC EXPOSED INDIVIDUALS (WITH AND WITHOUT SKIN LESIONS) AND IN SKIN CANCER TISSUES: A CASE CONTROL STUDY. CHRONIC ARSENIC TOXICITY HAS BECOME A GLOBAL CONCERN DUE TO ITS ADVERSE PATHOPHYSIOLOGICAL OUTCOME AND CARCINOGENIC POTENTIAL. IT IS ALREADY ESTABLISHED THAT ARSENIC INDUCED REACTIVE OXYGEN SPECIES ALTERS MITOCHONDRIAL FUNCTIONALITY. MAJOR REGULATORY GENES FOR MITOCHONDRIAL BIOGENESIS, I.E., PGC1ALPHA, TFAM, NRF1AND NRF2 ARE LOCATED IN THE NUCLEUS. AS A RESULT, MITOCHONDRIA-NUCLEUS CROSSTALK IS CRUCIAL FOR PROPER MITOCHONDRIAL FUNCTION. THIS PREVIOUS HYPOTHESIS LED US TO INVESTIGATEINVOLVEMENT OF EPIGENETIC ALTERATION BEHINDENHANCED MITOCHONDRIAL BIOGENESIS IN CHRONIC ARSENIC EXPOSURE. AN EXTENSIVE CASE-CONTROL STUDY WAS CONDUCTED WITH 390 STUDY PARTICIPANTS (UNEXPOSED, EXPOSED WITHOUT SKIN LESION, EXPOSED WITH SKIN LESION AND EXPOSED SKIN TUMOUR) FROM HIGHLY ARSENIC EXPOSED AREAS OFWEST BENGAL, INDIA. METHYLATION SPECIFIC PCRREVEALED SIGNIFICANT PROMOTER HYPOMETHYLATION OFTWO KEY BIOGENESIS REGULATORY GENES, PGC1ALPHAANDTFAM IN ARSENIC EXPOSED INDIVIDUALS AND ALSO IN SKIN TUMOUR TISSUES. LINEAR REGRESSION ANALYSIS INDICATED SIGNIFICANT NEGATIVE CORRELATION BETWEEN URINARY ARSENIC CONCENTRATION AND PROMOTER METHYLATION STATUS. INCREASED EXPRESSION OF BIOGENESIS REGULATORY GENES WASOBTAINED BY QUANTITATIVE REAL-TIME PCR ANALYSIS. MOREOVER, ALTERED MITOCHONDRIAL FUSION-FISSION REGULATORY GENE EXPRESSION WAS ALSO OBSERVED IN SKIN TUMOUR TISSUES. MIR663, HAVING TUMOUR SUPPRESSOR GENE LIKE FUNCTION WAS KNOWN TO BE EPIGENETICALLY REGULATED THROUGH MITOCHONDRIAL RETROGRADE SIGNAL. PROMOTER HYPERMETHYLATION WITH SIGNIFICANTLY DECREASED EXPRESSION OF MIR663 WAS FOUND IN SKIN CANCER TISSUES COMPARED TO NON-CANCEROUS CONTROL TISSUE. IN CONCLUSION, RESULTS INDICATED CRUCIAL ROLE OF EPIGENETIC ALTERATION IN ARSENIC INDUCED MITOCHONDRIAL BIOGENESIS AND ARSENICAL SKIN CARCINOGENESIS FOR THE FIRST TIME. HOWEVER, FURTHER MECHANISTIC STUDIES ARE NECESSARY FOR DETAILED UNDERSTANDING OF MITOCHONDRIA-NUCLEUS CROSSTALK IN ARSENIC PERTURBATION.	2020	
                                                                                                                                                                                                                                                                                                                                                                                        
6  984  29 CHRONIC PSYCHOLOGICAL STRESS ALTERS GENE EXPRESSION IN RAT COLON EPITHELIAL CELLS PROMOTING CHROMATIN REMODELING, BARRIER DYSFUNCTION AND INFLAMMATION. CHRONIC STRESS IS COMMONLY ASSOCIATED WITH ENHANCED ABDOMINAL PAIN (VISCERAL HYPERSENSITIVITY), BUT THE CELLULAR MECHANISMS UNDERLYING HOW CHRONIC STRESS INDUCES VISCERAL HYPERSENSITIVITY ARE POORLY UNDERSTOOD. IN THIS STUDY, WE EXAMINED CHANGES IN GENE EXPRESSION IN COLON EPITHELIAL CELLS FROM A RAT MODEL USING RNA-SEQUENCING TO EXAMINE STRESS-INDUCED CHANGES TO THE TRANSCRIPTOME. FOLLOWING CHRONIC STRESS, THE MOST SIGNIFICANTLY UP-REGULATED GENES INCLUDED ATG16L1, COQ10B, DCAF13, NAT2, PTBP2, RRAS2, SPINK4 AND DOWN-REGULATED GENES INCLUDING ABAT, CITED2, CNNM2, DAB2IP, PLEKHM1, SCD2, AND TAB2. THE PRIMARY ALTERED BIOLOGICAL PROCESSES REVEALED BY NETWORK ENRICHMENT ANALYSIS WERE INFLAMMATION/IMMUNE RESPONSE, TISSUE MORPHOGENESIS AND DEVELOPMENT, AND NUCLEOSOME/CHROMATIN ASSEMBLY. THE MOST SIGNIFICANTLY DOWN-REGULATED PROCESS WAS THE DIGESTIVE SYSTEM DEVELOPMENT/FUNCTION, WHEREAS THE MOST SIGNIFICANTLY UP-REGULATED PROCESSES WERE INFLAMMATORY RESPONSE, ORGANISMAL INJURY, AND CHROMATIN REMODELING MEDIATED BY H3K9 METHYLATION. FURTHERMORE, A SUBPOPULATION OF STRESSED RATS DEMONSTRATED VERY SIGNIFICANTLY ALTERED GENE EXPRESSION AND TRANSCRIPT ISOFORMS, ENRICHED FOR THE DIFFERENTIAL EXPRESSION OF GENES INVOLVED IN THE INFLAMMATORY RESPONSE, INCLUDING UPREGULATION OF CYTOKINE AND CHEMOKINE RECEPTOR GENE EXPRESSION COUPLED WITH DOWNREGULATION OF EPITHELIAL ADHERENS AND TIGHT JUNCTION MRNAS. IN SUMMARY, THESE FINDINGS SUPPORT THAT CHRONIC STRESS IS ASSOCIATED WITH INCREASED LEVELS OF CYTOKINES AND CHEMOKINES, THEIR DOWNSTREAM SIGNALING PATHWAYS COUPLED TO DYSREGULATION OF INTESTINAL CELL DEVELOPMENT AND FUNCTION. EPIGENETIC REGULATION OF CHROMATIN REMODELING LIKELY PLAYS A PROMINENT ROLE IN THIS PROCESS. RESULTS ALSO SUGGEST THAT SUPER ENHANCERS PLAY A PRIMARY ROLE IN CHRONIC STRESS-ASSOCIATED INTESTINAL BARRIER DYSFUNCTION.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
7 3437  29 HYPERGLYCEMIC MICROENVIRONMENT COMPROMISES THE HOMEOSTASIS OF COMMUNICATION BETWEEN THE BONE-BRAIN AXIS BY THE EPIGENETIC REPRESSION OF THE OSTEOCALCIN RECEPTOR, GPR158 IN THE HIPPOCAMPUS. DIABETES MELLITUS (DM) IS A CHRONIC METABOLIC DISEASE, MAINLY CHARACTERIZED BY INCREASED BLOOD GLUCOSE AND INSULIN DYSFUNCTION. IN RESPONSE TO THE PERSISTENT SYSTEMIC HYPERGLYCEMIC STATE, NUMEROUS METABOLIC AND PHYSIOLOGICAL COMPLICATIONS HAVE ALREADY BEEN WELL CHARACTERIZED. HOWEVER, ITS RELATIONSHIP TO BONE FRAGILITY, COGNITIVE DEFICITS AND INCREASED RISK OF DEMENTIA STILL NEEDS TO BE BETTER UNDERSTOOD. THE IMPACT OF CHRONIC HYPERGLYCEMIA ON BONE PHYSIOLOGY AND ARCHITECTURE WAS ASSESSED IN A MODEL OF CHRONIC HYPERGLYCEMIA INDUCED BY A SINGLE INTRAPERITONEAL ADMINISTRATION OF STREPTOZOTOCIN (STZ; 55 MG/KG) IN WISTAR RATS. IN ADDITION, THE BONE-TO-BRAIN COMMUNICATION WAS INVESTIGATED BY ANALYZING THE GENE EXPRESSION AND METHYLATION STATUS OF GENES THAT ENCODE THE MAIN OSTEOKINES RELEASED BY THE BONE [FGF23 (FIBROBLAST GROWTH FACTOR 23), BGLAP (BONE GAMMA-CARBOXYGLUTAMATE PROTEIN) AND LCN2 (LIPOCALIN 2) AND THEIR RECEPTORS IN BOTH, THE BONE AND THE BRAIN [FGFR1 (FIBROBLAST GROWTH FACTOR RECEPTOR 1), GPR6A (G-PROTEIN COUPLED RECEPTOR FAMILY C GROUP 6 MEMBER A), GPR158 (G PROTEIN-COUPLED RECEPTOR 158) AND SLC22A17 (SOLUTE CARRIER FAMILY 22 MEMBER 17)]. IT WAS OBSERVED THAT CHRONIC HYPERGLYCEMIA NEGATIVELY IMPACTED ON BONE BIOLOGY AND COMPROMISED THE BALANCE OF THE BONE-BRAIN ENDOCRINE AXIS. ULTRASTRUCTURAL DISORGANIZATION WAS ACCOMPANIED BY GLOBAL DNA HYPOMETHYLATION AND CHANGES IN GENE EXPRESSION OF DNA-MODIFYING ENZYMES THAT WERE ACCOMPANIED BY CHANGES IN THE METHYLATION STATUS OF THE OSTEOKINE PROMOTER REGION BGLAP AND LCN2 (LIPOCALIN 2) IN THE FEMUR. ADDITIONALLY, THE CHRONIC HYPERGLYCEMIC STATE WAS ACCOMPANIED BY MODULATION OF GENE EXPRESSION OF THE OSTEOKINES FGF23 (FIBROBLAST GROWTH FACTOR 23), BGLAP (BONE GAMMA-CARBOXYGLUTAMATE PROTEIN) AND LCN2 (LIPOCALIN 2) IN THE DIFFERENT BRAIN REGIONS. HOWEVER, TRANSCRIPTIONAL REGULATION MEDIATED BY DNA METHYLATION WAS OBSERVED ONLY FOR THE OSTEOKINE RECEPTORS, FGFR1(FIBROBLAST GROWTH FACTOR RECEPTOR 1) IN THE STRIATUM AND GPR158 (G PROTEIN-COUPLED RECEPTOR 158) IN THE HIPPOCAMPUS. THIS IS A PIONEER STUDY DEMONSTRATING THAT THE CHRONIC HYPERGLYCEMIC STATE COMPROMISES THE CROSSTALK BETWEEN BONE TISSUE AND THE BRAIN, MAINLY AFFECTING THE HIPPOCAMPUS, THROUGH TRANSCRIPTIONAL SILENCING OF THE BGLAP RECEPTOR BY HYPERMETHYLATION OF GPR158 GENE.	2023	

8 3049  25 GENOME-WIDE ANALYSIS REVEALS ZINC TRANSPORTER ZIP9 REGULATED BY DNA METHYLATION PROMOTES RADIATION-INDUCED SKIN FIBROSIS VIA THE TGF-BETA SIGNALING PATHWAY. RADIATION-INDUCED SKIN FIBROSIS IS A DETRIMENTAL AND CHRONIC DISORDER THAT OCCURS AFTER RADIATION EXPOSURE. DNA METHYLATION HAS BEEN CHARACTERIZED AS AN IMPORTANT REGULATORY MECHANISM OF MULTIPLE PATHOLOGICAL PROCESSES. IN THIS STUDY, WE COMPARED THE GENOME-WIDE DNA METHYLATION STATUS IN RADIATION-INDUCED FIBROTIC SKIN AND ADJACENT NORMAL TISSUES OF RATS BY METHYLATED DNA IMMUNOPRECIPITATION SEQUENCING. RADIATION-INDUCED FIBROTIC SKIN SHOWED DIFFERENTIALLY METHYLATED REGIONS ASSOCIATED WITH 3,650 PROTEIN-CODING GENES, 72 MICRORNAS, 5,836 LONG NONCODING RNAS AND 3 PIWI-INTERACTING RNAS. BY INTEGRATING THE MRNA AND METHYLATION PROFILES, THE ZINC TRANSPORTER SLC39A9/ZIP9 WAS INVESTIGATED IN GREATER DETAIL. THE PROTEIN LEVEL OF ZIP9 WAS INCREASED IN IRRADIATED SKIN TISSUES OF HUMANS, MONKEYS, AND RATS, ESPECIALLY IN RADIOGENIC FIBROTIC SKIN TISSUES. RADIATION INDUCED THE DEMETHYLATION OF A CPG DINUCLEOTIDE IN EXON 1 OF ZIP9 THAT RESULTED IN RECRUITMENT OF THE TRANSCRIPTIONAL FACTOR SP1 AND INCREASED ZIP9 EXPRESSION. OVEREXPRESSION OF ZIP9 RESULTED IN ACTIVATION OF THE PROFIBROTIC TRANSFORMING GROWTH FACTOR-BETA SIGNALING PATHWAY THROUGH PROTEIN KINASE B IN HUMAN FIBROBLASTS. IN ADDITION, RADIATION-INDUCED SKIN FIBROSIS WAS ASSOCIATED WITH INCREASED ZINC ACCUMULATION. THE ZINC CHELATOR N,N,N',N'-TETRAKIS(2-PYRIDYLMETHYL)-1,2-ETHYLENEDIAMINE ABROGATED ZIP9-INDUCED ACTIVATION OF THE TRANSFORMING GROWTH FACTOR-BETA SIGNALING PATHWAY AND ATTENUATED RADIATION-INDUCED SKIN FIBROSIS IN A RAT MODEL. IN SUMMARY, OUR FINDINGS ILLUSTRATE EPIGENETIC REGULATION OF ZIP9 AND ITS CRITICAL ROLE IN PROMOTING RADIATION-INDUCED SKIN FIBROSIS.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
9 5468  25 RESISTANCE TRAINING AND REDOX HOMEOSTASIS: CORRELATION WITH AGE-ASSOCIATED GENOMIC CHANGES. REGULAR PHYSICAL ACTIVITY IS EFFECTIVE AS PREVENTION AND TREATMENT FOR DIFFERENT CHRONIC CONDITIONS RELATED TO THE AGEING PROCESSES. IN FACT, A SEDENTARY LIFESTYLE HAS BEEN LINKED TO A WORSENING OF CELLULAR AGEING BIOMARKERS SUCH AS TELOMERE LENGTH (TL) AND/OR SPECIFIC EPIGENETIC CHANGES (E.G. DNA METHYLATION), WITH INCREASE OF THE PROPENSITY TO AGING-RELATED DISEASES AND PREMATURE DEATH. EXTENDING OUR PREVIOUS FINDINGS, WE AIMED TO TEST THE HYPOTHESIS THAT 12 WEEKS OF LOW FREQUENCY, MODERATE INTENSITY, EXPLOSIVE-TYPE RESISTANCE TRAINING (EMRT) MAY ATTENUATE AGE-ASSOCIATED GENOMIC CHANGES. TO THIS AIM, TL, GLOBAL DNA METHYLATION, TRF2, KU80, SIRT1, SIRT2 AND GLOBAL PROTEIN ACETYLATION, AS WELL AS OTHER PROTEINS INVOLVED IN APOPTOTIC PATHWAY (BCL-2, BAX AND CASPASE-3), ANTIOXIDANT RESPONSE (TRXR1 AND MNSOD) AND OXIDATIVE DAMAGE (MYELOPEROXIDASE) WERE EVALUATED BEFORE AND AFTER EMRT IN WHOLE BLOOD OR PERIPHERAL MONONUCLEAR CELLS (PBMCS) OF ELDERLY SUBJECTS. OUR FINDINGS CONFIRM THE POTENTIAL OF EMRT TO INDUCE AN ADAPTIVE CHANGE IN THE ANTIOXIDANT PROTEIN SYSTEMS AT SYSTEMIC LEVEL AND SUGGEST A PUTATIVE ROLE OF RESISTANCE TRAINING IN THE REDUCTION OF GLOBAL DNA METHYLATION. MOREOVER, WE OBSERVED THAT EMRT COUNTERACTS THE TELOMERES' SHORTENING IN A MANNER THAT PROVED TO BE DIRECTLY CORRELATED WITH THE AMELIORATION OF REDOX HOMEOSTASIS AND EFFICACY OF TRAINING REGIME, EVALUATED AS IMPROVEMENT OF BOTH MUSCLE'S POWER/STRENGTH AND FUNCTIONAL PARAMETERS.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
10 1966  30 EPIGENETIC ALTERATION OF PRKCDBP IN COLORECTAL CANCERS AND ITS IMPLICATION IN TUMOR CELL RESISTANCE TO TNFALPHA-INDUCED APOPTOSIS. PURPOSE: PRKCDBP IS A PUTATIVE TUMOR SUPPRESSOR IN WHICH ALTERATION HAS BEEN OBSERVED IN SEVERAL HUMAN CANCERS. WE INVESTIGATED EXPRESSION AND FUNCTION OF PRKCDBP IN COLORECTAL CELLS AND TISSUES TO EXPLORE ITS CANDIDACY AS A SUPPRESSOR IN COLORECTAL TUMORIGENESIS. EXPERIMENTAL DESIGN: EXPRESSION AND METHYLATION STATUS OF PRKCDBP AND ITS EFFECT ON TUMOR GROWTH WERE EVALUATED. TRANSCRIPTIONAL REGULATION BY NF-KAPPAB SIGNALING WAS DEFINED BY LUCIFERASE REPORTER AND CHROMATIN IMMUNOPRECIPITATION ASSAYS. RESULTS: PRKCDBP EXPRESSION WAS HARDLY DETECTABLE IN 29 OF 80 (36%) PRIMARY TUMORS AND 11 OF 19 (58%) CELL LINES, AND ITS ALTERATION CORRELATED WITH TUMOR STAGE AND GRADE. PROMOTER HYPERMETHYLATION WAS COMMONLY FOUND IN CANCERS. PRKCDBP EXPRESSION INDUCED THE G(1) CELL-CYCLE ARREST AND INCREASED CELLULAR SENSITIVITY TO VARIOUS APOPTOTIC STRESSES. PRKCDBP WAS INDUCED BY TNFALPHA, AND ITS LEVEL CORRELATED WITH TUMOR CELL SENSITIVITY TO TNFALPHA-INDUCED APOPTOSIS. PRKCDBP INDUCTION BY TNFALPHA WAS DISRUPTED BY BLOCKING NF-KAPPAB SIGNALING WHILE IT WAS ENHANCED BY RELA TRANSFECTION. THE PRKCDBP PROMOTER ACTIVITY WAS INCREASED IN RESPONSE TO TNFALPHA, AND THIS RESPONSE WAS ABOLISHED BY DISRUPTION OF A KAPPAB SITE IN THE PROMOTER. PRKCDBP DELAYED THE FORMATION AND GROWTH OF XENOGRAFT TUMORS AND IMPROVED TUMOR RESPONSE TO TNFALPHA-INDUCED APOPTOSIS. CONCLUSIONS: PRKCDBP IS A PROAPOPTOTIC TUMOR SUPPRESSOR WHICH IS COMMONLY ALTERED IN COLORECTAL CANCER BY PROMOTER HYPERMETHYLATION, AND ITS GENE TRANSCRIPTION IS DIRECTLY ACTIVATED BY NF-KAPPAB IN RESPONSE TO TNFALPHA. THIS SUGGESTS THAT PRKCDBP INACTIVATION MAY CONTRIBUTE TO TUMOR PROGRESSION BY REDUCING CELLULAR SENSITIVITY TO TNFALPHA AND OTHER STRESSES, PARTICULARLY UNDER CHRONIC INFLAMMATORY MICROENVIRONMENT.	2011	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
11 5205  28 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
12  217  24 ACUTE EXERCISE INCREASES THE EXPRESSION OF KIR2DS4 BY PROMOTER DEMETHYLATION IN NK CELLS. POSITIVE EFFECTS OF EXERCISE ON CANCER PREVENTION AND PROGRESSION HAVE BEEN PROPOSED TO BE MEDIATED BY STIMULATING NATURAL KILLER (NK) CELLS. BECAUSE NK CELL RECEPTORS ARE REGULATED BY EPIGENETIC MODIFICATIONS, WE INVESTIGATED WHETHER ACUTE AEROBIC EXERCISE AND TRAINING CHANGE PROMOTER DNA METHYLATION AND GENE EXPRESSION OF THE ACTIVATING KIR2DS4 AND THE INHIBITING KIR3DL1 GENE. SIXTEEN HEALTHY WOMEN (50-60 YEARS) PERFORMED A GRADED EXERCISE TEST (GXT) AND WERE RANDOMIZED INTO EITHER A PASSIVE CONTROL GROUP OR AN INTERVENTION GROUP PERFORMING A FOUR-WEEK ENDURANCE EXERCISE INTERVENTION. BLOOD SAMPLES (PRE-, POST-GXT AND POST-TRAINING) WERE USED FOR ISOLATION OF DNA/RNA OF NK CELLS TO ASSESS DNA PROMOTER METHYLATION BY TARGETED DEEP-AMPLICON SEQUENCING AND GENE EXPRESSION BY QRT-PCR. POTENTIAL CHANGES IN NK CELL SUBSETS WERE DETERMINED BY FLOW CYTOMETRY. ACUTE AND CHRONIC EXERCISE DID NOT PROVOKE SIGNIFICANT ALTERATIONS OF NK CELL PROPORTIONS. PROMOTER METHYLATION DECREASED AND GENE EXPRESSION INCREASED FOR KIR2DS4 AFTER ACUTE EXERCISE. A HIGH GENE EXPRESSION CORRELATED WITH A LOW METHYLATION OF CPGS THAT WERE ALTERED BY ACUTE EXERCISE. CHRONIC EXERCISE RESULTED IN A MINOR DECREASE OF DNA METHYLATION AND DID NOT ALTER GENE EXPRESSION. ACUTE EXERCISE PROVOKES EPIGENETIC MODIFICATIONS, AFFECTING THE BALANCE BETWEEN THE ACTIVATING KIR2DS4 AND THE INHIBITING KIR3DL1, WITH POTENTIAL BENEFITS ON NK CELL FUNCTION.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
13 3473  28 IDENTIFICATION OF A NOVEL, METHYLATION-DEPENDENT, RUNX2 REGULATORY REGION ASSOCIATED WITH OSTEOARTHRITIS RISK. OSTEOARTHRITIS (OA) IS A COMMON, MULTIFACTORIAL AND POLYGENIC SKELETAL DISEASE THAT, IN ITS SEVEREST FORM, REQUIRES JOINT REPLACEMENT SURGERY TO RESTORE MOBILITY AND TO RELIEVE CHRONIC PAIN. USING TISSUES FROM THE ARTICULATING JOINTS OF 260 PATIENTS WITH OA AND A RANGE OF IN VITRO EXPERIMENTS, INCLUDING CRISPR-CAS9, WE HAVE CHARACTERIZED AN INTERGENIC REGULATORY ELEMENT. HERE, GENOTYPE AT AN OA RISK LOCUS CORRELATES WITH DIFFERENTIAL DNA METHYLATION, WITH ALTERED GENE EXPRESSION OF BOTH A TRANSCRIPTIONAL REGULATOR (RUNX2), AND A CHROMATIN REMODELLING PROTEIN (SUPT3H). RUNX2 IS A STRONG CANDIDATE FOR OA SUSCEPTIBILITY, WITH ITS ENCODED PROTEIN BEING ESSENTIAL FOR SKELETOGENESIS AND HEALTHY JOINT FUNCTION. THE OA RISK LOCUS INCLUDES SINGLE NUCLEOTIDE POLYMORPHISMS (SNPS) LOCATED WITHIN AND FLANKING THE DIFFERENTIALLY METHYLATED REGION (DMR). THE OA ASSOCIATION SNP, RS10948172, DEMONSTRATES PARTICULARLY STRONG CORRELATION WITH METHYLATION, AND TWO INTERGENIC SNPS FALLING WITHIN THE DMR (RS62435998 AND RS62435999) DEMONSTRATE GENETIC AND EPIGENETIC EFFECTS ON THE REGULATORY ACTIVITY OF THIS REGION. WE THEREFORE POSIT THAT THE OA SIGNAL MEDIATES ITS EFFECT BY MODULATING THE METHYLATION OF THE REGULATORY ELEMENT, WHICH THEN IMPACTS ON GENE EXPRESSION, WITH RUNX2 BEING THE PRINCIPAL TARGET. OUR STUDY HIGHLIGHTS THE INTERPLAY BETWEEN DNA METHYLATION, OA GENETIC RISK AND THE DOWNSTREAM REGULATION OF GENES CRITICAL TO NORMAL JOINT FUNCTION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
14 2395  34 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
15 3306  27 HIGH-PHOSPHATE-INDUCED CALCIFICATION IS RELATED TO SM22ALPHA PROMOTER METHYLATION IN VASCULAR SMOOTH MUSCLE CELLS. HYPERPHOSPHATEMIA IS CLOSELY RELATED TO VASCULAR CALCIFICATION IN PATIENTS WITH CHRONIC KIDNEY DISEASE. VASCULAR SMOOTH MUSCLE CELLS (VSMCS) EXPOSED TO HIGH PHOSPHATE CONCENTRATIONS IN VITRO UNDERGO PHENOTYPIC TRANSITION TO OSTEOBLAST-LIKE CELLS. MECHANISMS UNDERLYING THIS TRANSDIFFERENTIATION ARE NOT CLEAR. IN THIS STUDY WE USED TWO IN VITRO MODELS, HUMAN AORTIC SMOOTH MUSCLE CELLS AND RAT AORTIC RINGS, TO INVESTIGATE THE PHENOTYPIC TRANSITION OF VSMCS INDUCED BY HIGH PHOSPHATE. WE FOUND THAT HIGH PHOSPHATE CONCENTRATION (3.3 MMOL/L) IN THE MEDIUM WAS ASSOCIATED WITH INCREASED DNA METHYLTRANSFERASE ACTIVITY AND METHYLATION OF THE PROMOTER REGION OF SM22ALPHA. THIS WAS ACCOMPANIED BY LOSS OF THE SMOOTH MUSCLE CELL-SPECIFIC PROTEIN SM22ALPHA, GAIN OF THE OSTEOBLAST TRANSCRIPTION FACTOR CBFA1, AND INCREASED ALKALINE PHOSPHATASE ACTIVITY WITH THE SUBSEQUENT IN VITRO CALCIFICATION. THE ADDITION OF A DEMETHYLATING AGENT (PROCAINE) TO THE HIGH-PHOSPHATE MEDIUM REDUCED DNA METHYLTRANSFERASE ACTIVITY AND PREVENTED METHYLATION OF THE SM22ALPHA PROMOTER, WHICH WAS ACCOMPANIED BY AN INCREASE IN SM22ALPHA EXPRESSION AND LESS CALCIFICATION. ADDITIONALLY, DOWNREGULATION OF SM22ALPHA, EITHER BY SIRNA OR BY A METHYL GROUP DONOR (S-ADENOSYL METHIONINE), RESULTED IN OVEREXPRESSION OF CBFA1. IN CONCLUSION, WE DEMONSTRATE THAT METHYLATION OF SM22ALPHA PROMOTER IS AN IMPORTANT EVENT IN VASCULAR SMOOTH MUSCLE CELL CALCIFICATION AND THAT HIGH PHOSPHATE INDUCES THIS EPIGENETIC MODIFICATION. THESE FINDINGS UNCOVER A NEW INSIGHT INTO MECHANISMS BY WHICH HIGH PHOSPHATE CONCENTRATION PROMOTES VASCULAR CALCIFICATION.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
16 4875  30 OVEREXPRESSION OF AKT1 ENHANCES ADIPOGENESIS AND LEADS TO LIPOMA FORMATION IN ZEBRAFISH. BACKGROUND: OBESITY IS A COMPLEX, MULTIFACTORIAL DISORDER INFLUENCED BY THE INTERACTION OF GENETIC, EPIGENETIC, AND ENVIRONMENTAL FACTORS. OBESITY INCREASES THE RISK OF CONTRACTING MANY CHRONIC DISEASES OR METABOLIC SYNDROME. RESEARCHERS HAVE ESTABLISHED SEVERAL MAMMALIAN MODELS OF OBESITY TO STUDY ITS UNDERLYING MECHANISM. HOWEVER, A LOWER VERTEBRATE MODEL FOR CONVENIENTLY PERFORMING DRUG SCREENING AGAINST OBESITY REMAINS ELUSIVE. THE SPECIFIC AIM OF THIS STUDY WAS TO CREATE A ZEBRAFISH OBESITY MODEL BY OVER EXPRESSING THE INSULIN SIGNALING HUB OF THE AKT1 GENE. METHODOLOGY/PRINCIPAL FINDINGS: SKIN ONCOGENIC TRANSFORMATION SCREENING SHOWS THAT A STABLE ZEBRAFISH TRANSGENIC OF TG(KRT4HSA.MYRAKT1)(CY18) DISPLAYS SEVERELY OBESE PHENOTYPES AT THE ADULT STAGE. IN TG(KRT4:HSA.MYRAKT1)(CY18), THE EXPRESSION OF EXOGENOUS HUMAN CONSTITUTIVELY ACTIVE AKT1 (MYRAKT1) CAN ACTIVATE ENDOGENOUS DOWNSTREAM TARGETS OF MTOR, GSK-3ALPHA/BETA, AND 70S6K. DURING THE EMBRYONIC TO LARVAL TRANSITORY PHASE, THE SPECIFIC OVER EXPRESSION OF MYRAKT1 IN SKIN CAN PROMOTE HYPERTROPHIC AND HYPERPLASTIC GROWTH. FROM 21 HOUR POST-FERTILIZATION (HPF) ONWARDS, MYRAKT1 TRANSGENE WAS ECTOPICALLY EXPRESSED IN SEVERAL MESENCHYMAL DERIVED TISSUES. THIS MAY BE THE RESULT OF THE INTEGRATION POSITION EFFECT. TG(KRT4:HSA.MYRAKT1)(CY18) CAUSED A RAPID INCREASE OF BODY WEIGHT, HYPERPLASTIC GROWTH OF ADIPOCYTES, ABNORMAL ACCUMULATION OF FAT TISSUES, AND BLOOD GLUCOSE INTOLERANCE AT THE ADULT STAGE. REAL-TIME RT-PCR ANALYSIS SHOWED THE MAJORITY OF KEY GENES ON REGULATING ADIPOGENESIS, ADIPOCYTOKINE, AND INFLAMMATION ARE HIGHLY UPREGULATED IN TG(KRT4:HSA.MYRAKT1)(CY18). IN CONTRAST, THE MYOGENESIS- AND SKELETOGENESIS-RELATED GENE TRANSCRIPTS ARE SIGNIFICANTLY DOWNREGULATED IN TG(KRT4:HSA.MYRAKT1)(CY18), SUGGESTING THAT EXCESS ADIPOCYTE DIFFERENTIATION OCCURS AT THE EXPENSE OF OTHER MESENCHYMAL DERIVED TISSUES. CONCLUSION/SIGNIFICANCE: COLLECTIVELY, THE FINDINGS OF THIS STUDY PROVIDE DIRECT EVIDENCE THAT AKT1 SIGNALING PLAYS AN IMPORTANT ROLE IN BALANCING NORMAL LEVELS OF FAT TISSUE IN VIVO. THE OBESE ZEBRAFISH EXAMINED IN THIS STUDY COULD BE A NEW POWERFUL MODEL TO SCREEN NOVEL DRUGS FOR THE TREATMENT OF HUMAN OBESITY.	2012	
                                                                                                                                                                                                                     
17 1504  27 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
18  148  32 ABERRANT FLUID SHEAR STRESS CONTRIBUTES TO ARTICULAR CARTILAGE PATHOGENESIS VIA EPIGENETIC REGULATION OF ZBTB20 BY H3K4ME3. PURPOSE: OSTEOARTHRITIS (OA) IS A COMMON DISEASE FOR HUMAN BEINGS, CHARACTERIZED BY SEVERE INFLAMMATION, CARTILAGE DEGRADATION, AND SUBCHONDRAL BONE DESTRUCTION. HOWEVER, CURRENT THERAPIES ARE LIMITED TO RELIEVING PAIN OR JOINT REPLACEMENT AND NO EFFECTIVE TREATMENT METHODS HAVE BEEN DISCOVERED TO IMPROVE DEGENERATIVE CHANGES. CURRENTLY, A VARIETY OF EVIDENCES HAVE INDICATED THAT ABERRANT MECHANICAL STIMULI IS CLOSELY ASSOCIATED WITH ARTICULAR JOINT PATHOGENESIS, WHILE THE DETAILED UNDERLYING MECHANISM REMAINS UNELUCIDATED. IN THE PRESENT STUDY, WE DETERMINED TO INVESTIGATE THE IMPACT OF EXCESSIVE HIGH FLUID SHEAR STRESS (FSS) ON PRIMARY CHONDROCYTES AND THE UNDERLYING EPIGENETIC MECHANISMS. MATERIALS AND METHODS: PHALLOIDIN STAINING AND EDU STAINING WERE USED TO EVALUATE CELL MORPHOLOGY AND VIABILITY. THE MRNA LEVEL AND PROTEIN LEVEL OF GENES WERE DETERMINED BY QPCR, WESTERN BLOT ASSAY, AND IMMUNOFLUORESCENCE STAINING. MECHANISTIC INVESTIGATION WAS PERFORMED THROUGH RNA-SEQUENCING AND CUT&TAG SEQUENCING. IN VIVO, WE ADOPTED UNILATERAL ANTERIOR CROSSBITES (UAC) MICE MODEL TO INVESTIGATE THE EXPRESSION OF H3K4ME3 AND ZBTB20 IN ABERRANT FORCE-RELATED CARTILAGE PATHOGENESIS. RESULTS: THE RESULTS DEMONSTRATED THAT FSS GREATLY DISRUPTS CELL MORPHOLOGY AND SIGNIFICANTLY DECREASED CHONDROCYTE VIABILITY. ABERRANT FSS INDUCES REMARKABLE INFLAMMATORY MEDIATORS PRODUCTION, LEADING TO CARTILAGE DEGENERATION AND DEGRADATION. IN DEPTH MECHANISTIC STUDY SHOWED THAT FSS RESULTS IN MORE THAN 10-FOLD UPREGULATION OF H3K4ME3, AND THE MODULATORY EFFECT OF H3K4ME3 ON CARTILAGE WAS OBTAINED BY DIRECTLY TARGETING ZBTB20. FURTHERMORE, WNT SIGNALING WAS STRONGLY ACTIVATED IN HIGH FSS-INDUCED OA PATHOGENESIS, AND THE NEGATIVE IMPACT OF ZBTB20 ON CHONDROCYTES WAS ALSO ACHIEVED THROUGH ACTIVATING WNT SIGNALING PATHWAY. MOREOVER, PHARMACOLOGICAL INHIBITION OF H3K4ME3 ACTIVATION BY MM-102 OR TREATMENT WITH WNT PATHWAY INHIBITOR LF3 COULD EFFECTIVELY ALLEVIATE THE DESTRUCTIVE EFFECT OF FSS ON CHONDROCYTES. IN VIVO UAC MICE MODEL VALIDATED THE DYSREGULATION OF H3K4ME3 AND ZBTB20 IN ABERRANT FORCE-INDUCED CARTILAGE PATHOGENESIS. CONCLUSION: THROUGH THE COMBINATION OF IN VITRO FSS MODEL AND IN VIVO UAC MODEL, KMT2B-H3K4ME3-ZBTB20 AXIS WAS FIRST IDENTIFIED IN ABERRANT FSS-INDUCED CARTILAGE PATHOGENESIS, WHICH MAY PROVIDE EVIDENCES FOR EPIGENETIC-BASED THERAPY IN THE FUTURE.	2021	
     
19 4528  23 MULTIGENERATIONAL EFFECTS OF CADMIUM ON THE LIFESPAN AND FERTILITY OF DROSOPHILA MELANOGASTER. ALTHOUGH THE DAMAGE AND TOLERANCE MECHANISMS OF CD STRESS ARE KNOWN, THE DATA ON GENETIC RISK ARE LIMITED. THE AIM OF THIS STUDY WAS TO ASSESS THE CHRONIC TOXICITY OF CD, GENETIC RESPONSES, AND MULTIGENERATIONAL EFFECTS IN FIVE GENERATIONS OF DROSOPHILA MELANOGASTER. FOR EACH GENERATION, LIFESPAN AND FERTILITY WERE STATISTICALLY ANALYSED AND THE EXPRESSION OF APOPTOSIS- (P53 AND CASPASE-3) AND EPIGENESIS-RELATED (DDNMT2 AND DMBD2/3) GENES WAS EXAMINED. LIFESPAN AND FERTILITY SIGNIFICANTLY DECLINED UNDER CD STRESS AND THESE EFFECTS WERE MAINTAINED FOR TWO GENERATIONS AND ONE GENERATION, RESPECTIVELY, WHEN CD STRESS WAS REMOVED. THE EXPRESSION OF P53 AND CASPASE-3 WAS SIGNIFICANTLY UP-REGULATED AFTER EXPOSURE, SUGGESTING THAT APOPTOSIS CONTRIBUTES TO THE RESISTANCE MECHANISM. THEIR ALTERED EXPRESSION WAS RETAINED FOR TWO GENERATIONS. FURTHERMORE, HIGH EXPRESSION OF DDNMT2 AND DMBD2/3 ACCOMPANIED CD EXPOSURE, WHICH WAS PASSED ON TO THREE GENERATIONS, SUGGESTING THAT GENETIC MODIFICATIONS IN APOPTOSIS-RELATED GENES ARE CARRIED TO THE OFFSPRING THROUGH EPIGENETIC REGULATION.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
20 1011  28 CIGARETTE SMOKE CONDENSATE INDUCES DIFFERENTIAL EXPRESSION AND PROMOTER METHYLATION PROFILES OF CRITICAL GENES INVOLVED IN LUNG CANCER IN NL-20 LUNG CELLS IN VITRO: SHORT-TERM AND CHRONIC EXPOSURE. ESTABLISHING EARLY DIAGNOSTIC MARKERS OF HARM IS CRITICAL FOR EFFECTIVE PREVENTION PROGRAMS AND REGULATION OF TOBACCO PRODUCTS. THIS STUDY EXAMINED EFFECTS OF CIGARETTE SMOKE CONDENSATE (CSC) ON EXPRESSION AND PROMOTER METHYLATION PROFILE OF CRITICAL GENES (DAPK, ECAD, MGMT, AND RASSF1A) INVOLVED IN LUNG CANCER DEVELOPMENT IN DIFFERENT HUMAN LUNG CELL LINES. NL-20 CELLS WERE TREATED WITH 0.1-100 MUG/ML OF CSC FOR 24 TO 72 HRS FOR SHORT-TERM EXPOSURES. DAPK EXPRESSION OR METHYLATION STATUS WAS NOT SIGNIFICANTLY AFFECTED. HOWEVER, CSC TREATMENT RESULTED IN CHANGES IN EXPRESSION AND PROMOTER METHYLATION PROFILE OF ECAD, MGMT, AND RASSF1A. FOR CHRONIC STUDIES, CELLS WERE EXPOSED TO 1 OR 10 MUG/ML CSC UP TO 28 DAYS. CELLS SHOWED MORPHOLOGICAL CHANGES ASSOCIATED WITH TRANSFORMATION AND CHANGES IN INVASION CAPACITIES AND GLOBAL METHYLATION STATUS. THIS STUDY PROVIDES CRITICAL DATA SUGGESTING THAT EPIGENETIC CHANGES COULD SERVE AS AN EARLY BIOMARKER OF HARM DUE TO EXPOSURE TO CIGARETTE SMOKE.	2013