1 5864 108 SUPPRESSION OF ELF4 IN ULCERATIVE COLITIS PREDISPOSES HOST TO COLORECTAL CANCER. ULCERATIVE COLITIS (UC) IS A CHRONIC INFLAMMATORY BOWEL DISEASE, CHARACTERIZED BY RELAPSING AND REMITTING COLON MUCOSAL INFLAMMATION. FOR PATIENTS SUFFERING FROM UC, A HIGHER RISK OF COLON CANCER HAS BEEN WIDELY RECOGNIZED. HERE, WE FOUND THAT ELF4 (-/-) MICE DEVELOPED COLON TUMORS WITH 3 CYCLES OF DEXTRAN SULFATE SODIUM SALT (DSS) TREATMENT ALONE. WE FURTHER SHOWED THAT ELF4 SUPPRESSION WAS PREVALENT IN BOTH PATIENTS WITH UC AND DSS-INDUCED MICE MODELS, AND THIS SUPPRESSION WAS CAUSED BY PROMOTER REGION METHYLATION. ELF4, UPON PARYLATION BY PARP1, TRANSCRIPTIONALLY REGULATED MULTIPLE DNA DAMAGE REPAIR MACHINERY COMPONENTS. CONSISTENTLY, ELF4 DEFICIENCY LEADS TO MORE SEVERE DNA DAMAGE BOTH IN VITRO AND IN VIVO. ORAL ADMINISTRATION OF MONTMORILLONITE POWDER CAN PREVENT THE REDUCTION OF ELF4 IN DSS-INDUCED COLITIS MODELS AND LOWER THE RISK OF COLON TUMOR DEVELOPMENT DURING AZOXYMETHANE (AOM) AND DSS INDUCED COLITIS-ASSOCIATED CANCER (CAC). THESE DATA PROVIDED ADDITIONAL MECHANISM OF CAC INITIATION AND SUPPORTED THE "EPIGENETIC PRIMING MODEL OF TUMOR INITIATION". 2021 2 5963 31 TEM STUDY OF CHRONIC ALCOHOLISM EFFECTS ON EARLY CARCINOGENESIS BY PROBING THE NANOSCALE STRUCTURAL ALTERATIONS OF CELL NUCLEI. NANOSCALE STRUCTURAL ALTERATION IN THE NUCLEI OF CELLS WITH THE PROGRESSION OF CARCINOGENESIS IS DUE TO THE REARRANGEMENTS OF THE BASIC BUILDING BLOCKS IN THE CELL SUCH AS DNA, RNA, LIPIDS, ETC. ALTHOUGH EPIGENETIC MODIFICATIONS UNDERLIE THE DEVELOPMENT OF CANCER, EXPOSURE TO CARCINOGENIC CHEMICALS SUCH AS ALCOHOL ALSO ENHANCES THE DEVELOPMENT OF CANCER. WE REPORT THE EFFECTS OF CHRONIC ALCOHOLISM ON EARLY-CARCINOGENESIS BASED ON CHANGES IN THE DEGREE OF NANOSCALE STRUCTURAL ALTERATIONS (L (D)) IN NUCLEI. FOR THIS, TRANSMISSION ELECTRON MICROSCOPY (TEM) IMAGING OF THE NUCLEI OF COLONIC CELLS IS PERFORMED FOR THE FOLLOWING FOUR MOUSE MODELS: CONTROL MICE; CHRONIC ALCOHOLIC MICE TREATED WITH ETHANOL (I.E., ETOH MICE); MICE TREATED WITH COLONIC CARCINOGEN AZOXYMETHANE (AOM) AND DEXTRAN SULFATE SODIUM (DSS) THAT INDUCED COLITIS (I.E., AOM + DSS MICE); AND CHRONIC ALCOHOLIC OR ETOH TREATED MICE, TOGETHER WITH AOM AND DSS TREATMENT (I.E., AOM + DSS + ETOH MICE). THE DISORDERED OPTICAL LATTICES ARE CONSTRUCTED FROM THEIR RESPECTIVE TEM IMAGES OF THIN COLONIC CELL NUCLEI AND THE L (D) VALUES ARE CALCULATED USING THE INVERSE PARTICIPATION RATIO (IPR) TECHNIQUE FROM THE SPATIALLY LOCALIZED EIGENFUNCTIONS OF THESE LATTICES. RESULTS SHOW NO SIGNIFICANT DIFFERENCE IN THE AVERAGE L (D) VALUE OF THE COLON CELL NUCLEI OF ALCOHOL TREATED MICE RELATIVE TO ITS CONTROL [I.E., L (D)(C) APPROXIMATELY L (D)(ETOH)]; HOWEVER, AN INCREASE IN THE L (D) VALUE OF ALCOHOL TREATED PRECANCEROUS CELLS [I.E., L (D)(AOM + DSS + ETOH) > L (D)(AOM + DSS)], INDICATING THAT ALCOHOL ACCELERATES THE EARLY CARCINOGENIC PROCESS. 2021 3 5999 27 THE ACETYLOME REGULATORS HDAC1 AND HDAC2 DIFFERENTLY MODULATE INTESTINAL EPITHELIAL CELL DEPENDENT HOMEOSTATIC RESPONSES IN EXPERIMENTAL COLITIS. HISTONE DEACETYLASES (HDAC) REMOVE ACETYL GROUPS FROM PROTEINS, INFLUENCING GLOBAL AND SPECIFIC GENE EXPRESSION. HDACS CONTROL INFLAMMATION, AS SHOWN BY HDAC INHIBITOR-DEPENDENT PROTECTION FROM DEXTRAN SULFATE SODIUM (DSS)-INDUCED MURINE COLITIS. ALTHOUGH TISSUE-SPECIFIC HDAC KNOCKOUTS SHOW REDUNDANT AND SPECIFIC FUNCTIONS, LITTLE IS KNOWN OF THEIR INTESTINAL EPITHELIAL CELL (IEC) ROLE. WE HAVE SHOWN PREVIOUSLY THAT DUAL HDAC1/HDAC2 IEC-SPECIFIC LOSS DISRUPTS CELL PROLIFERATION AND DETERMINATION, WITH DECREASED SECRETORY CELL NUMBERS AND ALTERED BARRIER FUNCTION. WE THUS INVESTIGATED HOW COMPOUND HDAC1/HDAC2 OR HDAC2 IEC-SPECIFIC DEFICIENCY ALTERS THE INFLAMMATORY RESPONSE. FLOXED HDAC1 AND HDAC2 AND VILLIN-CRE MICE WERE INTERBRED. COMPOUND HDAC1/HDAC2 IEC-DEFICIENT MICE SHOWED CHRONIC BASAL INFLAMMATION, WITH INCREASED BASAL DISEASE ACTIVITY INDEX (DAI) AND DEREGULATED REG GENE COLONIC EXPRESSION. DSS-TREATED DUAL HDAC1/HDAC2 IEC-DEFICIENT MICE DISPLAYED INCREASED DAI, HISTOLOGICAL SCORE, INTESTINAL PERMEABILITY, AND INFLAMMATORY GENE EXPRESSION. IN CONTRAST TO DOUBLE KNOCKOUTS, HDAC2 IEC-SPECIFIC LOSS DID NOT AFFECT IEC DETERMINATION AND GROWTH, NOR RESULT IN CHRONIC INFLAMMATION. HOWEVER, HDAC2 DISRUPTION PROTECTED AGAINST DSS COLITIS, AS SHOWN BY DECREASED DAI, INTESTINAL PERMEABILITY AND CASPASE-3 CLEAVAGE. HDAC2 IEC-SPECIFIC DEFICIENT MICE DISPLAYED INCREASED EXPRESSION OF IEC GENE SUBSETS, SUCH AS COLONIC ANTIMICROBIAL REG3B AND REG3G MRNAS, AND DECREASED EXPRESSION OF IMMUNE CELL FUNCTION-RELATED GENES. OUR DATA SHOW THAT HDAC1 AND HDAC2 ARE ESSENTIAL IEC HOMEOSTASIS REGULATORS. IEC-SPECIFIC HDAC1 AND HDAC2 MAY ACT AS EPIGENETIC SENSORS AND TRANSMITTERS OF ENVIRONMENTAL CUES AND REGULATE IEC-MEDIATED MUCOSAL HOMEOSTATIC AND INFLAMMATORY RESPONSES. DIFFERENT LEVELS OF IEC HDAC ACTIVITY MAY LEAD TO POSITIVE OR NEGATIVE OUTCOMES ON INTESTINAL HOMEOSTASIS DURING INFLAMMATION. 2014 4 6118 37 THE EPIGENETIC EFFECTS OF ASPIRIN: THE MODIFICATION OF HISTONE H3 LYSINE 27 ACETYLATION IN THE PREVENTION OF COLON CARCINOGENESIS IN AZOXYMETHANE- AND DEXTRAN SULFATE SODIUM-TREATED CF-1 MICE. COLORECTAL CANCER (CRC) IS THE THIRD MOST COMMON CANCER WORLDWIDE. CHRONIC INFLAMMATION APPEARS TO ENHANCE THE RISK OF CRC. EMERGING EVIDENCE HAS SUGGESTED THAT EPIGENETIC MECHANISMS PLAY AN IMPORTANT ROLE IN CRC. ASPIRIN [ACETYLSALICYLIC ACID (ASA)] HAS BEEN SHOWN TO PREVENT CRC; HOWEVER, THE EPIGENETIC MECHANISMS OF ITS ACTION REMAIN UNKNOWN. THIS STUDY INVESTIGATED THE PROTECTIVE ROLE OF ASA IN AZOXYMETHANE (AOM)-INITIATED AND DEXTRAN SULFATE SODIUM (DSS)-PROMOTED COLITIS-ASSOCIATED COLON CANCER (CAC) AND EXAMINED THE EPIGENETIC EFFECTS, PARTICULARLY ON HISTONE 3 LYSINE 27 ACETYLATION (H3K27AC), UNDERLYING THE PREVENTIVE EFFECT OF ASA. CF-1 MICE WERE FED WITH AIN-93M DIET WITH OR WITHOUT 0.02% ASA FROM 1 WEEK PRIOR TO AOM INITIATION UNTIL THE MICE WERE KILLED 20 WEEKS AFTER AOM INJECTION. OUR RESULTS SHOWED THAT AOM/DSS + ASA SIGNIFICANTLY SUPPRESSED INFLAMMATORY COLITIS SYMPTOMS AND TUMOR MULTIPLICITY. AOM/DSS + ASA REDUCED AOM/DSS-INDUCED PROTEIN EXPRESSION AND THE ACTIVITY OF HISTONE DEACETYLASES (HDACS) AND GLOBALLY RESTORED H3K27AC. FURTHERMORE, AOM/DSS + ASA INHIBITED AOM/DSS-INDUCED ENRICHMENT OF H3K27AC IN THE PROMOTERS OF INDUCIBLE NITRIC OXIDE SYNTHASE (INOS), TUMOR NECROSIS FACTOR ALPHA (TNF-ALPHA) AND INTERLEUKIN 6 (IL-6) THAT CORRESPONDED TO THE DRAMATIC SUPPRESSION OF THE MESSENGER RNA (MRNA) AND PROTEIN LEVELS. SURPRISINGLY, NO SIGNIFICANT CHANGES IN THE H3K27AC ABUNDANCE IN THE PROSTAGLANDIN-ENDOPEROXIDE SYNTHASE 2 (COX-2) PROMOTERS OR IN THE COX-2 MRNA AND PROTEIN EXPRESSION WERE OBSERVED. COLLECTIVELY, OUR RESULTS SUGGEST THAT A POTENTIAL NOVEL EPIGENETIC MECHANISM UNDERLIES THE CHEMOPREVENTIVE EFFECTS OF ASA, AND THIS MECHANISM ATTENUATES CAC IN AOM/DSS-INDUCED CF-1 MICE VIA THE INHIBITION OF HDACS AND THE MODIFICATION OF H3K27AC MARKS THAT SUPPRESS INOS, TNF-ALPHA AND IL-6. 2016 5 1905 39 ENHANCER OF ZESTE HOMOLOG 2 CONTRIBUTES TO APOPTOSIS BY INACTIVATING JANUS KINASE 2/ SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION SIGNALING IN INFLAMMATORY BOWEL DISEASE. BACKGROUND: INFLAMMATORY BOWEL DISEASE (IBD) IS A PREVALENT WORLDWIDE HEALTH PROBLEM FEATURED BY RELAPSING, CHRONIC GASTROINTESTINAL INFLAMMATION. ENHANCER OF ZESTE HOMOLOG 2 (EZH2) IS A CRITICAL EPIGENETIC REGULATOR IN DIFFERENT PATHOLOGICAL MODELS, SUCH AS CANCER AND INFLAMMATION. HOWEVER, THE ROLE OF EZH2 IN THE IBD DEVELOPMENT IS STILL OBSCURE. AIM: TO EXPLORE THE EFFECT OF EZH2 ON IBD PROGRESSION AND THE UNDERLYING MECHANISM. METHODS: THE IBD MOUSE MODEL WAS CONDUCTED BY ADDING DEXTRAN SODIUM SULFATE (DSS), AND THE EFFECT OF EZH2 ON DSS-INDUCED COLITIS WAS ASSESSED IN THE MODEL. THE FUNCTION OF EZH2 IN REGULATING APOPTOSIS AND PERMEABILITY WAS EVALUATED BY ANNEXIN V-FITC APOPTOSIS DETECTION KIT, TRANSEPITHELIAL ELECTRICAL RESISTANCE ANALYSIS, AND WESTERN BLOT ANALYSIS OF RELATED MARKERS, INCLUDING ZONA OCCLUDENS 1, CLAUDIN-5, AND OCCLUDIN, IN NCM460 AND FETAL HUMAN COLON (FHC) CELLS. THE MECHANICAL INVESTIGATION WAS PERFORMED BY QUANTITATIVE REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION, WESTERN BLOT ANALYSIS, AND CHROMATIN IMMUNOPRECIPITATION ASSAYS. RESULTS: THE COLON LENGTH WAS INHIBITED IN THE DSS-TREATED MICE AND WAS ENHANCED BY THE EZH2 DEPLETION IN THE SYSTEM. DSS TREATMENT CAUSED A DECREASED HISTOLOGICAL SCORE IN THE MICE, WHICH WAS REVERSED BY EZH2 DEPLETION. THE INFLAMMATORY CYTOKINES, SUCH AS TUMOR NECROSIS FACTOR-ALPHA, INTERLEUKIN-6, AND INTERLEUKIN-1BETA, WERE INDUCED IN THE DSS-TREATED MICE, IN WHICH THE DEPLETION OF EZH2 COULD REVERSE THIS EFFECT. MOREOVER, THE TUMOR NECROSIS FACTOR-ALPHA TREATMENT INDUCED THE APOPTOSIS OF NCM460 AND FHC CELLS, IN WHICH EZH2 DEPLETION COULD REVERSE THIS EFFECT IN THE CELLS. MOREOVER, THE DEPLETION OF EZH2 ATTENUATED PERMEABILITY OF COLONIC EPITHELIAL CELLS. MECHANICALLY, THE DEPLETION OF EZH2 OR EZH2 INHIBITOR GSK343 WAS ABLE TO ENHANCE THE EXPRESSION AND THE PHOSPHORYLATION OF JANUS KINASE 2 (JK2) AND SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION IN THE NCM460 AND FHC CELLS. SPECIFICALLY, EZH2 INACTIVATED JAK2 EXPRESSION BY REGULATING HISTONE H3K27ME3. JAK2 INHIBITOR TG101348 WAS ABLE TO REVERSE EZH2 KNOCKDOWN-MEDIATED COLONIC EPITHELIAL CELL PERMEABILITY AND APOPTOSIS. CONCLUSION: THUS, WE CONCLUDED THAT EZH2 CONTRIBUTED TO APOPTOSIS AND INFLAMMATORY RESPONSE BY INACTIVATING JAK2/ SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION SIGNALING IN IBD. EZH2 MAY BE APPLIED AS A POTENTIAL TARGET FOR IBD THERAPY. 2021 6 3867 30 JMJD3/H3K27ME3 EPIGENETIC MODIFICATION REGULATES TH17/TREG CELL DIFFERENTIATION IN ULCERATIVE COLITIS. ULCERATIVE COLITIS (UC) IS A CHRONIC NONSPECIFIC INFLAMMATORY BOWEL DISEASE CHARACTERIZED BY CHRONIC INFLAMMATION AND ULCERATION OF THE COLONIC MUCOSA, FREQUENT RELAPSE, AND CANCERIZATION THAT IS DIFFICULT TO CURE. IN RECENT YEARS, THE INCIDENCE OF UC HAS INCREASED. HOWEVER, ITS ETIOLOGY AND PATHOGENESIS ARE STILL NOT COMPLETELY CLEAR. IN THIS STUDY, DEXTRAN SODIUM SULFATE (DSS) WAS USED TO INDUCE THE MODEL, AND GSK-J1 AND DEXAMETHASONE WERE ADMINISTERED TO THE MICE. A VARIETY OF MOLECULAR BIOLOGY AND IMMUNOLOGICAL TECHNIQUES, SUCH AS IMMUNOFLUORESCENCE, PCR AND CHROMATIN IMMUNOPRECIPITATION (CHIP), WERE USED TO EXAMINE JMJD3/H3K27ME3-MEDIATED REGULATION OF TH17/TREG CELL DIFFERENTIATION IN UC BY TARGETING HISTONE MODIFICATION. THIS STUDY WILL PROVIDE AN IMPORTANT THEORETICAL BASIS FOR UNDERSTANDING THE PATHOGENESIS AND POTENTIAL THERAPEUTIC TARGETS OF UC. 2022 7 5794 26 STAT1 EPIGENETICALLY REGULATES LCP2 AND TNFAIP2 BY RECRUITING EP300 TO CONTRIBUTE TO THE PATHOGENESIS OF INFLAMMATORY BOWEL DISEASE. BACKGROUND: THE AETIOLOGY OF INFLAMMATORY BOWEL DISEASE (IBD) IS RELATED TO GENETICS AND EPIGENETICS. EPIGENETIC REGULATION OF THE PATHOGENESIS OF IBD HAS NOT BEEN WELL DEFINED. HERE, WE INVESTIGATED THE ROLE OF H3K27AC EVENTS IN THE PATHOGENESIS OF IBD. BASED ON PREVIOUS CHIP-SEQ AND RNA-SEQ ASSAYS, WE STUDIED SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION 1 (STAT1) AS A TRANSCRIPTION FACTOR (TF) AND INVESTIGATED WHETHER THE STAT1-EP300-H3K27AC AXIS CONTRIBUTES TO THE DEVELOPMENT OF IBD. WE PERFORMED CHIP-PCR TO INVESTIGATE THE INTERACTION BETWEEN STAT1 AND H3K27AC, AND CO-IP ASSAYS WERE PERFORMED TO INVESTIGATE THE CROSSTALK BETWEEN STAT1 AND EP300. RESULTS: LYMPHOCYTE CYTOSOLIC PROTEIN 2 (LCP2) AND TNF-ALPHA-INDUCIBLE PROTEIN 2 (TNFAIP2) ARE TARGET GENES OF STAT1. P-STAT1 BINDS TO THE ENHANCER LOCI OF THE TWO GENES WHERE H3K27AC IS ENRICHED, AND EP300 SUBSEQUENTLY BINDS TO REGULATE THEIR EXPRESSION. IN MICE WITH DEXTRAN SULFATE SODIUM (DSS)-INDUCED ACUTE COLITIS, AN EP300 INHIBITOR SIGNIFICANTLY INHIBITED COLITIS. CONCLUSIONS: P-STAT1 AND EP300 PROMOTE TNFAIP2 AND LCP2 EXPRESSION THROUGH AN INCREASE IN H3K27AC ENRICHMENT ON THEIR ENHANCERS AND CONTRIBUTE TO THE PATHOGENESIS OF CHRONIC INFLAMMATION. 2021 8 4933 29 PATERNAL CHRONIC COLITIS CAUSES EPIGENETIC INHERITANCE OF SUSCEPTIBILITY TO COLITIS. INFLAMMATORY BOWEL DISEASE (IBD) ARISES BY UNKNOWN ENVIRONMENTAL TRIGGERS IN GENETICALLY SUSCEPTIBLE INDIVIDUALS. EPIGENETIC REGULATION OF GENE EXPRESSION MAY INTEGRATE INTERNAL AND EXTERNAL INFLUENCES AND MAY THEREBY MODULATE DISEASE SUSCEPTIBILITY. EPIGENETIC MODIFICATION MAY ALSO AFFECT THE GERM-LINE AND IN CERTAIN CONTEXTS CAN BE INHERITED TO OFFSPRING. THIS STUDY INVESTIGATES EPIGENETIC ALTERATIONS CONSEQUENT TO EXPERIMENTAL MURINE COLITIS INDUCED BY DEXTRAN SODIUM SULPHATE (DSS), AND THEIR PATERNAL TRANSMISSION TO OFFSPRING. GENOME-WIDE METHYLOME- AND TRANSCRIPTOME-PROFILING OF INTESTINAL EPITHELIAL CELLS (IECS) AND SPERM CELLS OF MALES OF THE F0 GENERATION, WHICH RECEIVED EITHER DSS AND CONSEQUENTLY DEVELOPED COLITIS (F0(DSS)), OR NON-SUPPLEMENTED TAP WATER (F0(CTRL)) AND HENCE REMAINED HEALTHY, AND OF THEIR F1 OFFSPRING WAS PERFORMED USING REDUCED REPRESENTATION BISULFITE SEQUENCING (RRBS) AND RNA-SEQUENCING (RNA-SEQ), RESPECTIVELY. OFFSPRING OF F0(DSS) MALES EXHIBITED ABERRANT METHYLATION AND EXPRESSION PATTERNS OF MULTIPLE GENES, INCLUDING IGF1R AND NR4A2, WHICH ARE INVOLVED IN ENERGY METABOLISM. IMPORTANTLY, DSS COLITIS IN F0(DSS) MICE WAS ASSOCIATED WITH DECREASED BODY WEIGHT AT BASELINE OF THEIR F1 OFFSPRING, AND THESE F1 MICE EXHIBITED INCREASED SUSCEPTIBILITY TO DSS-INDUCED COLITIS COMPARED TO OFFSPRING FROM F0(CTRL) MALES. THIS STUDY HENCE DEMONSTRATES EPIGENETIC TRANSMISSIBILITY OF METABOLIC AND INFLAMMATORY TRAITS RESULTING FROM EXPERIMENTAL COLITIS. 2016 9 5601 31 RORALPHA IS CRUCIAL FOR ATTENUATED INFLAMMATORY RESPONSE TO MAINTAIN INTESTINAL HOMEOSTASIS. RETINOIC ACID-RELATED ORPHAN RECEPTOR ALPHA (RORALPHA) FUNCTIONS AS A TRANSCRIPTION FACTOR FOR VARIOUS BIOLOGICAL PROCESSES, INCLUDING CIRCADIAN RHYTHM, CANCER, AND METABOLISM. HERE, WE GENERATE INTESTINAL EPITHELIAL CELL (IEC)-SPECIFIC RORALPHA-DEFICIENT (RORALPHA(DELTAIEC)) MICE AND FIND THAT RORALPHA IS CRUCIAL FOR MAINTAINING INTESTINAL HOMEOSTASIS BY ATTENUATING NUCLEAR FACTOR KAPPAB (NF-KAPPAB) TRANSCRIPTIONAL ACTIVITY. RORALPHA(DELTAIEC) MICE EXHIBIT EXCESSIVE INTESTINAL INFLAMMATION AND HIGHLY ACTIVATED INFLAMMATORY RESPONSES IN THE DEXTRAN SULFATE SODIUM (DSS) MOUSE COLITIS MODEL. TRANSCRIPTOME ANALYSIS REVEALS THAT DELETION OF RORALPHA LEADS TO UP-REGULATION OF NF-KAPPAB TARGET GENES IN IECS. CHROMATIN IMMUNOPRECIPITATION ANALYSIS REVEALS CORECRUITMENT OF RORALPHA AND HISTONE DEACETYLASE 3 (HDAC3) ON NF-KAPPAB TARGET PROMOTERS AND SUBSEQUENT DISMISSAL OF CREB BINDING PROTEIN (CBP) AND BROMODOMAIN-CONTAINING PROTEIN 4 (BRD4) FOR TRANSCRIPTIONAL REPRESSION. TOGETHER, WE DEMONSTRATE THAT RORALPHA/HDAC3-MEDIATED ATTENUATION OF NF-KAPPAB SIGNALING CONTROLS THE BALANCE OF INFLAMMATORY RESPONSES, AND THERAPEUTIC STRATEGIES TARGETING THIS EPIGENETIC REGULATION COULD BE BENEFICIAL TO THE TREATMENT OF CHRONIC INFLAMMATORY DISEASES, INCLUDING INFLAMMATORY BOWEL DISEASE (IBD). 2019 10 3658 35 INDUCTION OF ABERRANT TRIMETHYLATION OF HISTONE H3 LYSINE 27 BY INFLAMMATION IN MOUSE COLONIC EPITHELIAL CELLS. A FIELD FOR CANCERIZATION (FIELD DEFECT), WHERE GENETIC AND EPIGENETIC ALTERATIONS ARE ACCUMULATED IN NORMAL-APPEARING TISSUES, IS INVOLVED IN HUMAN CARCINOGENESIS, ESPECIALLY CANCERS ASSOCIATED WITH CHRONIC INFLAMMATION. ALTHOUGH ABERRANT DNA METHYLATION IS INVOLVED IN THE FIELD DEFECT AND INDUCED BY CHRONIC INFLAMMATION, IT IS STILL UNCLEAR FOR TRIMETHYLATION OF HISTONE H3 LYSINE 27 (H3K27ME3), WHICH IS INVOLVED IN GENE REPRESSION INDEPENDENT OF DNA METHYLATION AND FUNCTIONS AS A PRE-MARK FOR ABERRANT DNA METHYLATION. IN THIS STUDY, USING A MOUSE COLITIS MODEL INDUCED BY DEXTRAN SULFATE SODIUM (DSS), WE AIMED TO CLARIFY WHETHER ABERRANT H3K27ME3 IS INDUCED BY INFLAMMATION AND INVOLVED IN A FIELD DEFECT. CHIP-ON-CHIP ANALYSIS OF COLONIC EPITHELIAL CELLS REVEALED THAT H3K27ME3 LEVELS WERE INCREASED OR DECREASED FOR 266 GENOMIC REGIONS BY AGING, AND MORE EXTENSIVELY (23 INCREASED AND 3574 DECREASED REGIONS) BY COLITIS. SUCH INCREASE OR DECREASE OF H3K27ME3 WAS INDUCED AS EARLY AS 2 WEEKS AFTER THE INITIATION OF DSS TREATMENT, AND PERSISTED AT LEAST FOR 16 WEEKS EVEN AFTER THE INFLAMMATION DISAPPEARED. SOME OF THE ABERRANT H3K27ME3 IN COLONIC EPITHELIAL CELLS WAS CARRIED OVER INTO COLON TUMORS. FURTHERMORE, H3K27ME3 ACQUIRED AT DAPK1 BY COLITIS WAS FOLLOWED BY INCREASED DNA METHYLATION, SUPPORTING ITS FUNCTION AS A PRE-MARK FOR ABERRANT DNA METHYLATION. THESE RESULTS DEMONSTRATED THAT ABERRANT H3K27ME3 CAN BE INDUCED BY EXPOSURE TO A SPECIFIC ENVIRONMENT, SUCH AS COLITIS, AND SUGGESTED THAT ABERRANT HISTONE MODIFICATION, IN ADDITION TO ABERRANT DNA METHYLATION, IS INVOLVED IN THE FORMATION OF A FIELD DEFECT. 2012 11 4174 31 MELATONIN MEDIATED INHIBITION OF EZH2-NOS2 CROSSTALK ATTENUATES INFLAMMATORY BOWEL DISEASE IN PRECLINICAL IN VITRO AND IN VIVO MODELS. AIMS: INFLAMMATORY BOWEL DISEASE IS CHARACTERISED BY ABDOMINAL PAIN, DIARRHOEA, RECTAL BLEEDING AND WEIGHT LOSS. SOMETIMES IT MAY LEADS TO SEVERE HEALTH COMPLICATIONS RESULTING IN DEATH OF AN INDIVIDUAL. CURRENT RESEARCH EFFORTS TO HIGHLIGHT THE ROLE OF MELATONIN IN REGULATING EZH2, A MASTER EPIGENETIC REGULATOR AND ITS BENEFICIARY EFFECT IN CASE OF IBD MANAGEMENT. MATERIAL METHODS: MURINE MACROPHAGES (RAW 264.7) WERE TREATED WITH LIPOPOLYSACCHARIDES (LPS) TO ACTIVATE THEM FOR GENERATING INFLAMMATORY RESPONSE TO INVESTIGATE EFFICACY OF MELATONIN IN-VITRO MODELS. SIMILARLY, FOR DEVELOPING IN VIVO MODELS, DEXTRAN SODIUM SULPHATE (36-50 KDA) WAS USED. EVALUATIONS OF ANTI-INFLAMMATORY ACTIVITIES WERE CARRIED OUT BY NITRITE ASSAY, WESTERN BLOTTING, Q-PCR, IMMUNOFLUORESCENCE, AND HISTOLOGICAL STUDIES. KEY FINDINGS: REDUCTION OF EPIGENETIC TARGET, EZH2 BY MELATONIN SIGNIFICANTLY IMPROVES THE CLINICAL SYMPTOMS OF DEXTRAN SODIUM SULPHATE INDUCED COLITIS AND MAY BE IMPLICATED AS A POTENTIAL THERAPEUTIC TARGET IN IBD MANAGEMENT. THE PRESENT STUDY EVALUATES THE EFFICACY OF MELATONIN BY EPIGENETIC REGULATION IN IBD MODELS. DOWN REGULATION OF EZH2 BY MELATONIN REDUCED THE CHEMICAL INDUCED INFLAMMATORY INSULTS IN IN VITRO AND IN VIVO MODELS. EXPLORATION OF MOLECULAR PATHWAYS HAS REVEALED INTERLINK OF EZH2 AND NOS2, A HALLMARK OF INFLAMMATION. MOLECULAR MECHANISTIC ACTION OF MELATONIN IS ATTRIBUTED TO INHIBITION OF THE EXPRESSION AND PHYSICAL INTERACTION OF EZH2 AND NOS2. SIGNIFICANCE: OUR STUDY HIGHLIGHTS MELATONIN THERAPEUTIC EFFECT VIA ATTENUATING INTERACTION BETWEEN EZH2 AND NOS2 WHICH IS BENEFICIAL IN MANAGING IBD TREATMENT. 2022 12 2957 36 GENETIC AND EPIGENETIC IMPACT OF CHRONIC INFLAMMATION ON COLON MUCOSA CELLS. CHRONIC INFLAMMATION INCREASES CANCER RISK, AND CANCER DEVELOPMENT IS CHARACTERIZED BY STEPWISE ACCUMULATION OF GENETIC AND EPIGENETIC ALTERATIONS. DURING CHRONIC INFLAMMATION, INFECTIOUS AGENTS AND INTRINSIC MEDIATORS OF INFLAMMATORY RESPONSES CAN INDUCE GENETIC AND EPIGENETIC CHANGES. THIS STUDY TRIED TO EVALUATE BOTH THE GENETIC AND EPIGENETIC INFLUENCE OF CHRONIC INFLAMMATION ON COLON MUCOSA CELLS. REPETITIVE DEXTRAN SULFATE SODIUM (DSS) TREATMENT INDUCED CHRONIC COLITIS MODEL. WHOLE-EXOME SEQUENCING (WES) (200X COVERAGE) WAS PERFORMED TO DETECT SOMATIC VARIATIONS IN COLON MUCOSA CELLS. WITH THE USE OF WHOLE-GENOME BISULFITE SEQUENCING (BS) AT 34-FOLD COVERAGE (17-FOLD PER STRAND), THE METHYLOME OF BOTH THE COLITIS AND CONTROL TISSUE WAS COMPARATIVELY ANALYZED. BIOINFORMATICS ASSAY SHOWED THAT THERE WAS NO SIGNIFICANT SINGLE-NUCLEOTIDE POLYMORPHISM/INSERTION OR DELETION (SNP/INDEL) MUTATION ACCUMULATION IN COLITIS TISSUE, WHILE IT ACCUMULATED IN AGED MICE. FORTY-EIGHT GENES WITH SNP/INDEL MUTATION WERE OVERLAPPED IN THE THREE COLITIS TISSUES, TWO (WNT3A AND LAMA2) OF WHICH ARE IN THE CANCER DEVELOPMENT-RELATED SIGNALING PATHWAY. DIFFERENTIALLY METHYLATED REGION (DMR) ASSAY SHOWED THAT MANY GENES IN THE COLITIS TISSUE ARE ENRICHED IN THE CANCER DEVELOPMENT-RELATED SIGNALING PATHWAY, SUCH AS PI3K-AKT, RAS, WNT, TGF-BETA, AND MAPK SIGNALING PATHWAY. TOGETHER, THESE DATA SUGGESTED THAT EVEN THOUGH CHRONIC INFLAMMATION DID NOT OBVIOUSLY INCREASE GENETIC MUTATION ACCUMULATION, IT COULD BOTH GENETICALLY AND EPIGENETICALLY ALTER SOME GENES RELATED TO CANCER DEVELOPMENT. 2021 13 5602 35 RORGAMMAT(+) HEMATOPOIETIC CELLS ARE NECESSARY FOR TUMOR CELL PROLIFERATION DURING COLITIS-ASSOCIATED TUMORIGENESIS IN MICE. COLORECTAL CANCER (CRC) IS ONE OF THE MOST COMMON TUMOR ENTITIES. IN PATIENTS WITH INFLAMMATORY BOWEL DISEASES, THE DEVELOPMENT OF COLITIS-ASSOCIATED COLON CANCER IS CONSIDERED A DANGEROUS LONG-TERM COMPLICATION. IL-17A AND THE TRANSCRIPTION FACTOR RETINOIC ACID RECEPTOR-RELATED ORPHAN RECEPTOR GAMMAT (RORGAMMAT) PLAY FUNDAMENTAL ROLES IN THE PATHOGENESIS OF INFLAMMATORY BOWEL DISEASES; IN HUMAN STUDIES, WE DETECTED A DENSE INFILTRATION OF RORGAMMAT-DEPENDENT CD4(+) IL17A(+) T HELPER (TH)17 CELLS IN SPECIMENS OF CRC, ULCERATIVE COLITIS, AND ULCERATIVE COLITIS-ASSOCIATED COLORECTAL CANCER. HOWEVER, THE MECHANISTIC ROLE OF RORGAMMAT(+) HEMATOPOIETIC CELLS IN COLITIS-ASSOCIATED TUMORIGENESIS REMAINS UNCLEAR. TO INVESTIGATE COLITIS-ASSOCIATED COLON TUMORIGENESIS, WE CONDUCTED STUDIES IN THE AOM+DSS MOUSE MODEL THAT REVEALED THE IMPORTANCE OF RORGAMMAT FOR COLON TUMOR PROGRESSION. IN THE ABSENCE OF RORGAMMAT-DEPENDENT TH17 LYMPHOCYTES, MICE SHOWED SIGNS OF INTENSE CHRONIC COLITIS, BUT DEVELOPED SIGNIFICANTLY FEWER MACROSCOPIC TUMOR NODULES. THE REDUCTION OF TUMOR DEVELOPMENT IN RORGAMMAT(-/-) MICE WAS NOT DUE TO REDUCED COLON TUMOR INITIATION. HOWEVER, THE PROLIFERATION RATE OF TUMOR CELLS WAS REDUCED IN THE ABSENCE OF RORGAMMAT-DEPENDENT TH17 CELLS AND TUMOR CELLS SHOWED PRONOUNCED SIGNS OF SENESCENCE-ASSOCIATED EPIGENETIC AND LYSOSOMAL CHANGES. THESE RESULTS INDICATE AN IMPORTANT ROLE FOR THE IMMUNOLOGICAL MILIEU IN COLITIS-ASSOCIATED CANCER, WHICH IS SHAPED IN-PART BY RORGAMMAT-DEPENDENT TH17 LYMPHOCYTES THAT SUPPORT CRC GROWTH. 2015 14 2784 24 EZH2 PROMOTES EXTRACELLULAR MATRIX DEGRADATION VIA NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN PULPITIS. PULPITIS IS A COMPLICATED CHRONIC INFLAMMATORY PROCESS WHICH CAN BE IN A DYNAMIC BALANCE BETWEEN DAMAGE AND REPAIR. THE EXTRACELLULAR MATRIX PLAYS AN IMPORTANT REGULATORY ROLE IN WOUND HEALING AND TISSUE REPAIR. THE AIM OF THIS STUDY WAS TO EXPLORE THE ROLE OF THE EPIGENETIC MARK, ENHANCER OF ZESTE HOMOLOG 2 (EZH2) ON THE DEGRADATION OF EXTRACELLULAR MATRIX DURING PULPITIS. QUANTITATIVE POLYMERASE CHAIN REACTION WAS USED TO ASSESS THE EXPRESSION OF MATRIX METALLOPROTEINASES (MMPS) AND TYPE I COLLAGEN IN HUMAN DENTAL PULP CELLS (HDPCS) UPON EZH2 AND EI1 (EZH2 INHIBITOR) STIMULATION. THE MECHANISM OF EZH2 AFFECTING EXTRACELLULAR MATRIX WAS EXPLORED THROUGH QUANTITATIVE POLYMERASE CHAIN REACTION AND WESTERN BLOT. A RAT MODEL OF DENTAL PULP INFLAMMATION WAS ESTABLISHED, AND THE EXPRESSION OF TYPE I COLLAGEN IN DENTAL PULP UNDER EZH2 STIMULATION WAS DETECTED BY IMMUNOHISTOCHEMICAL STAINING. EZH2 UPREGULATED THE EXPRESSION OF MMP-1, MMP-3, MMP-8, AND MMP-10 AND DECREASED THE PRODUCTION OF TYPE I COLLAGEN IN HDPCS, WHILE EI1 HAD THE OPPOSITE EFFECT. EZH2 ACTIVATED THE NUCLEAR FACTOR-KAPPA B (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN HDPCS, THE INHIBITION OF WHICH REVERSED THE INDUCTION OF MMPS AND THE SUPPRESSION OF TYPE I COLLAGEN. EZH2 CAN DOWNREGULATE THE TYPE I COLLAGEN LEVELS IN AN EXPERIMENTAL MODEL OF DENTAL PULPITIS IN RATS. EZH2 PROMOTES EXTRACELLULAR MATRIX DEGRADATION VIA NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN PULPITIS. EZH2 CAN DECREASE THE TYPE I COLLAGEN LEVELS IN VIVO AND IN VITRO. 2021 15 3373 29 HISTONE MODULATION BLOCKS TREG-INDUCED FOXP3 BINDING TO THE IL-2 PROMOTER OF VIRUS-SPECIFIC CD8(+) T CELLS FROM FELINE IMMUNODEFICIENCY VIRUS-INFECTED CATS. CD8(+) T CELLS ARE CRITICAL FOR CONTROLLING HIV INFECTION. DURING THE CHRONIC PHASE OF LENTIVIRAL INFECTION, CD8(+) T CELLS LOSE THEIR PROLIFERATIVE CAPACITY AND EXHIBIT IMPAIRED ANTIVIRAL FUNCTION. THIS LOSS OF CD8(+) T CELL FUNCTION IS DUE, IN PART, TO CD4(+)CD25(+) T REGULATORY (TREG) CELL-MEDIATED SUPPRESSION. OUR RESEARCH GROUP HAS DEMONSTRATED THAT LENTIVIRUS-ACTIVATED CD4(+)CD25(+) TREG CELLS INDUCE THE REPRESSIVE TRANSCRIPTION FACTOR FORKHEAD BOX P3 (FOXP3) IN AUTOLOGOUS CD8(+) T CELLS FOLLOWING CO-CULTURE. WE HAVE RECENTLY REPORTED THAT TREG-INDUCED FOXP3 BINDS THE INTERLEUKIN-2 (IL-2), INTERFERON-GAMMA (IFN- GAMMA), AND TUMOR NECROSIS FACTOR-ALPHA (TNF-ALPHA) PROMOTERS IN VIRUS-SPECIFIC CD8(+) T CELLS. THESE DATA SUGGEST AN IMPORTANT ROLE OF FOXP3-MEDIATED CD8(+) T CELL DYSFUNCTION IN LENTIVIRAL INFECTION. TO ELUCIDATE THE MECHANISM OF THIS SUPPRESSION, WE PREVIOUSLY REPORTED THAT DECREASED METHYLATION FACILITATES FOXP3 BINDING IN MITOGEN-ACTIVATED CD8(+) T CELLS FROM FELINE IMMUNODEFICIENCY VIRUS (FIV)-INFECTED CATS. WE DEMONSTRATED THE REDUCED BINDING OF FOXP3 TO THE IL-2 PROMOTER BY INCREASING METHYLATION OF CD8(+) T CELLS. IN THE STUDIES PRESENTED HERE, WE ASK IF ANOTHER FORM OF EPIGENETIC MODULATION MIGHT ALLEVIATE FOXP3-MEDIATED SUPPRESSION IN CD8(+) T CELLS. WE HYPOTHESIZED THAT DECREASING HISTONE ACETYLATION IN VIRUS-SPECIFIC CD8(+) T CELLS WOULD DECREASE TREG-INDUCED FOXP3 BINDING TO THE IL-2 PROMOTER. INDEED, USING ANACARDIC ACID (AA), A KNOWN HISTONE ACETYL TRANSFERASE (HAT) INHIBITOR, WE DEMONSTRATE A REDUCTION IN FOXP3 BINDING TO THE IL-2 PROMOTER IN VIRUS-SPECIFIC CD8(+) T CELLS CO-CULTURED WITH AUTOLOGOUS TREG CELLS. THESE DATA IDENTIFY A NOVEL MECHANISM OF FOXP3-MEDIATED CD8(+) T CELL DYSFUNCTION DURING LENTIVIRAL INFECTION. 2018 16 1906 24 ENHANCER OF ZESTE HOMOLOG 2-CATALYSED H3K27 TRIMETHYLATION PLAYS A KEY ROLE IN ACUTE-ON-CHRONIC LIVER FAILURE VIA TNF-MEDIATED PATHWAY. ACUTE-ON-CHRONIC LIVER FAILURE IS MAINLY DUE TO HOST IMMUNITY SELF-DESTRUCTION. THE HISTONE H3 LYSINE 27 (H3K27) TRIMETHYLATING ENZYME, ENHANCER OF ZESTE HOMOLOG 2 (EZH2) MEDIATES EPIGENETIC SILENCING OF GENE EXPRESSION AND REGULATES IMMUNITY, ALSO INVOLVES PATHOGENESIS OF SEVERAL LIVER DISEASES. THE CURRENT STUDY WAS TO DETERMINE THE ROLE OF METHYLTRANSFERASE EZH2 AND ITS CATALYSED H3K27 TRIMETHYLATION (H3K27ME3) IN LIVER FAILURE, AND TO FURTHER INVESTIGATE THE POTENTIAL TARGET FOR LIVER FAILURE TREATMENT. EZH2 AND ITS CATALYSED H3K27ME3 WERE DETERMINED IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMC) FROM LIVER FAILURE PATIENTS AND KUPFFER CELLS FROM EXPERIMENTAL MICE. FURTHERMORE, GSK126 (AN INHIBITOR FOR EZH2 TRIMETHYLATION FUNCTION) WAS APPLIED IN LIVER FAILURE MICE IN VIVO, AND LIPOPOLYSACCHARIDE-STIMULATED MONONUCLEAR CELLS IN VITRO. EZH2 AND H3K27ME3 WERE SIGNIFICANTLY UPREGULATED IN HUMAN PBMC FROM LIVER FAILURE PATIENTS OR MURINE KUPFFER CELLS FROM THE LIVER FAILURE ANIMALS, RESPECTIVELY. GSK126 AMELIORATED DISEASE SEVERITY IN LIVER FAILURE MICE, WHICH MAYBE ATTRIBUTE TO DOWN-REGULATE CIRCULATING AND HEPATIC PROINFLAMMATORY CYTOKINES, ESPECIALLY TNF VIA REDUCING H3K27ME3. IN-DEPTH CHROMATIN IMMUNOPRECIPITATION ANALYSIS UNRAVELLED THAT DECREASED ENRICHMENT OF H3K27ME3 ON TNF PROMOTOR, RESULTING IN TNF ELEVATION IN KUPFFER CELLS FROM LIVER FAILURE MICE. NUCLEAR FACTOR KAPPA B (NF-KAPPAB) AND PROTEIN KINASE B (AKT) SIGNALLING PATHWAYS WERE ACTIVATED UPON LIPOPOLYSACCHARIDE STIMULATION, BUT ATTENUATED BY USING GSK126, ACCOMPANIED WITH DECREASED TNF IN VITRO. IN CONCLUSION, EZH2 AND H3K27ME3 CONTRIBUTED TO THE PATHOGENESIS OF LIVER FAILURE VIA TRIGGERING TNF AND OTHER INDISPENSABLE PROINFLAMMATORY CYTOKINES. EZH2 WAS TO MODIFY H3K27ME3 ENRICHMENT, AS WELL AS, ACTIVATION OF THE DOWNSTREAM NF-KAPPAB AND AKT SIGNALLING PATHWAYS. 2018 17 1945 25 EPIGALLOCATECHIN-3-GALLATE, A HISTONE ACETYLTRANSFERASE INHIBITOR, INHIBITS EBV-INDUCED B LYMPHOCYTE TRANSFORMATION VIA SUPPRESSION OF RELA ACETYLATION. BECAUSE THE P300/CBP-MEDIATED HYPERACETYLATION OF RELA (P65) IS CRITICAL FOR NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) ACTIVATION, THE ATTENUATION OF P65 ACETYLATION IS A POTENTIAL MOLECULAR TARGET FOR THE PREVENTION OF CHRONIC INFLAMMATION. DURING OUR ONGOING SCREENING STUDY TO IDENTIFY NATURAL COMPOUNDS WITH HISTONE ACETYLTRANSFERASE INHIBITOR (HATI) ACTIVITY, WE IDENTIFIED EPIGALLOCATECHIN-3-GALLATE (EGCG) AS A NOVEL HATI WITH GLOBAL SPECIFICITY FOR THE MAJORITY OF HAT ENZYMES BUT WITH NO ACTIVITY TOWARD EPIGENETIC ENZYMES INCLUDING HDAC, SIRT1, AND HMTASE. AT A DOSE OF 100 MICROMOL/L, EGCG ABROGATES P300-INDUCED P65 ACETYLATION IN VITRO AND IN VIVO, INCREASES THE LEVEL OF CYTOSOLIC IKAPPABALPHA, AND SUPPRESSES TUMOR NECROSIS FACTOR ALPHA (TNFALPHA)-INDUCED NF-KAPPAB ACTIVATION. WE ALSO SHOWED THAT EGCG PREVENTS TNFALPHA-INDUCED P65 TRANSLOCATION TO THE NUCLEUS, CONFIRMING THAT HYPERACETYLATION IS CRITICAL FOR NF-KAPPAB TRANSLOCATION AS WELL AS ACTIVITY. FURTHERMORE, EGCG TREATMENT INHIBITED THE ACETYLATION OF P65 AND THE EXPRESSION OF NF-KAPPAB TARGET GENES IN RESPONSE TO DIVERSE STIMULI. FINALLY, EGCG REDUCED THE BINDING OF P300 TO THE PROMOTER REGION OF INTERLEUKIN-6 GENE WITH AN INCREASED RECRUITMENT OF HDAC3, WHICH HIGHLIGHTS THE IMPORTANCE OF THE BALANCE BETWEEN HATS AND HISTONE DEACETYLASES IN THE NF-KAPPAB-MEDIATED INFLAMMATORY SIGNALING PATHWAY. IMPORTANTLY, EGCG AT 50 MICROMOL/L DOSE COMPLETELY BLOCKS EBV INFECTION-INDUCED CYTOKINE EXPRESSION AND SUBSEQUENTLY THE EBV-INDUCED B LYMPHOCYTE TRANSFORMATION. THESE RESULTS SHOW THE CRUCIAL ROLE OF ACETYLATION IN THE DEVELOPMENT OF INFLAMMATORY-RELATED DISEASES. 2009 18 5850 21 SUBEROYLANILIDE HYDROXAMIC ACID (SAHA) REDUCES FIBROSIS MARKERS AND DEACTIVATES HUMAN STELLATE CELLS VIA THE EPITHELIAL-MESENCHYMAL TRANSITION (EMT). HEPATIC FIBROSIS IS KNOWN AS THE ACCUMULATION OF CONNECTIVE TISSUE SECONDARY TO CHRONIC DAMAGE TO THE LIVER. EPITHELIAL-MESENCHYMAL TRANSITION (EMT) CORRESPONDING INCREASE IN LIVER FIBROGENESIS WAS SHOWN WITH IMMUNOHISTOCHEMISTRY AND PCR-BASED STUDIES. SUBEROYLANILIDE HYDROXAMIC ACID (SAHA), A SYNTHETIC COMPOUND APPROVED AS A HISTONE DEACETYLASE INHIBITOR (HDAC) BY THE FDA TO TREAT CUTANEOUS T-CELL LYMPHOMA IS UNDER INVESTIGATION FOR THE TREATMENT OF LUNG AND RENAL FIBROSIS. EXPERIMENTAL MODELING FOR HEPATIC FIBROSIS CAN BE CONSTRUCTED WITH AN LX2 CELL LINE ISOLATED FROM HUMAN HEPATIC STELLATE CELLS (HSCS). IN THIS STUDY, WE AIMED TO INVESTIGATE THE MODULATION OF SAHA IN THE PATHOGENESIS OF LIVER FIBROSIS BY DETECTING THE LEVELS OF PROTEINS; (E-CADHERIN (E-CAD), N-CADHERIN (N-CAD), VIMENTIN (VIM), AND GENES; E-CAD, N-CAD, VIM, TRANSFORMING GROWTH FACTOR-BETA (TGF-BETA), ALPHA-SMOOTH MUSCLE ACTIN (ALPHA-SMA), TYPE 1 COLLAGEN (COL1A1), TYPE 3 COLLAGEN (COL3A1)) THAT PLAY A SIGNIFICANT ROLE IN EMT WITH THE LX2 CELL LINE. WE ALSO EVALUATED THE ACTION OF SAHA WITH CELL PROLIFERATION, CLONOGENIC, AND MIGRATION ASSAY. CELL PROLIFERATION WAS PERFORMED BY FLOW CYTOMETRY. ALL THE PROTEIN LEVELS WERE DETERMINED BY WESTERN BLOT ANALYSIS, AND GENE EXPRESSION LEVELS WERE MEASURED BY REAL-TIME PCR. OUR STUDY OBSERVED THAT SAHA TREATMENT DECREASED CELL VIABILITY, COLONY FORMATION AND MIGRATION IN LX2 CELLS. WE FOUND THAT SAHA INCREASED E-CAD EXPRESSION LEVEL, WHILE IT DECREASED N-CAD, VIM, COL1A1, COL3A1, ALPHA-SMA TGF-BETA GENES EXPRESSION LEVELS. SAHA DECREASED THE LEVEL OF E-CAD, N-CAD, AND VIM PROTEIN LEVELS. WE THOUGHT THAT SAHA POSSESSES POTENT ANTIFIBROTIC AND ANTI-EMT PROPERTIES IN LX2. 2021 19 1455 22 DISCOVERY OF THIENO[2,3-D]PYRIMIDINE-BASED HYDROXAMIC ACID DERIVATIVES AS BROMODOMAIN-CONTAINING PROTEIN 4/HISTONE DEACETYLASE DUAL INHIBITORS INDUCE AUTOPHAGIC CELL DEATH IN COLORECTAL CARCINOMA CELLS. BROMODOMAIN-CONTAINING PROTEIN 4 (BRD4) AND HISTONE DEACETYLASES (HDAC) ARE BOTH ATTRACTIVE EPIGENETIC TARGETS IN CANCER AND OTHER CHRONIC DISEASES. BASED ON THE INTEGRATED FRAGMENT-BASED DRUG DESIGN, SYNTHESIS, AND IN VITRO AND IN VIVO EVALUATIONS, A SERIES OF NOVEL THIENO[2,3-D]PYRIMIDINE-BASED HYDROXAMIC ACID DERIVATIVES ARE DISCOVERED AS SELECTIVE BRD4-HDAC DUAL INHIBITORS. COMPOUND 17C IS THE MOST POTENT INHIBITOR FOR BRD4 AND HDAC WITH IC(50) VALUES AT NANOMOLAR LEVELS, AS WELL AS THE EXPRESSION LEVEL OF C-MYC, AND INCREASES THE ACETYLATION OF HISTONE H3. MOREOVER, 17C PRESENTS INHIBITORY EFFECTS ON THE PROLIFERATION OF COLORECTAL CARCINOMA (CRC) CELLS VIA INDUCING AUTOPHAGIC CELL DEATH. IT ALSO HAS A GOOD PHARMACOKINETIC PROFILE IN RATS AND ORAL BIOAVAILABILITY OF 40.5%. IN THE HCT-116 XENOGRAFT IN VIVO MODELS, 17C DISPLAYS POTENT INHIBITORY EFFICIENCY ON TUMOR GROWTH BY INDUCING AUTOPHAGIC CELL DEATH AND SUPPRESSING IL6-JAK-STAT SIGNALING PATHWAYS. OUR RESULTS SUGGEST THAT THE BRD4-HDAC DUAL INHIBITION MIGHT BE AN ATTRACTIVE THERAPEUTIC STRATEGY FOR CRC. 2020 20 6661 33 UPREGULATION OF DNA METHYLTRANSFERASE-MEDIATED GENE SILENCING, ANCHORAGE-INDEPENDENT GROWTH, AND MIGRATION OF COLON CANCER CELLS BY INTERLEUKIN-6. INFLAMMATORY BOWEL DISEASE IS CHARACTERIZED BY CHRONIC INFLAMMATION WHICH PREDISPOSES TO COLORECTAL CANCER. THE MECHANISMS BY WHICH INFLAMMATION PROMOTES TUMORIGENESIS ARE NOT FULLY KNOWN. WE AIMED TO INVESTIGATE THE LINKS BETWEEN COLONIC INFLAMMATION AND TUMORIGENESIS VIA EPIGENETIC GENE SILENCING. COLON CANCER SPECIMENS WERE ASSESSED FOR THE EXPRESSION OF DNA METHYLTRANSFERASE-1 (DNMT-1) USING IMMUNOHISTOCHEMISTRY. COLORECTAL CARCINOMA CELL LINES WERE ASSESSED FOR DNMT1 EXPRESSION, METHYLCYTOSINE CONTENT, PROMOTER METHYLATION, GENE EXPRESSION, AND TUMORIGENESIS IN RESPONSE TO INTERLEUKIN (IL)-6. DNMT1 WAS EXPRESSED AT HIGHER LEVELS IN BOTH THE PERITUMORAL STROMA AND TUMOR IN INFLAMMATORY BOWEL DISEASE-ASSOCIATED CANCERS COMPARED WITH SPORADIC COLON CANCERS. IL-6 TREATMENT OF COLON CANCER CELLS RESULTED IN AN INCREASE IN DNMT1 EXPRESSION, INDEPENDENT OF DE NOVO GENE EXPRESSION. IL-6 INCREASED THE METHYLATION OF PROMOTER REGIONS OF GENES ASSOCIATED WITH TUMOR SUPPRESSION, ADHESION, AND APOPTOSIS RESISTANCE. EXPRESSION OF A SUBSET OF THESE GENES WAS DOWNREGULATED BY IL-6, AN EFFECT THAT WAS PREVENTED BY PREINCUBATION WITH 5-AZADEOXYCYTIDINE, A DNMT1 INHIBITOR. ANCHORAGE-INDEPENDENT GROWTH AND MIGRATION OF COLON CANCER CELLS WAS ALSO INCREASED BY IL-6 IN A 5-AZADEOXYCYTIDINE-SENSITIVE MANNER. OUR RESULTS INDICATE THAT DNMT-MEDIATED GENE SILENCING MAY PLAY A ROLE IN INFLAMMATION-ASSOCIATED COLON TUMORIGENESIS. 2010