1 6599 110 TWIST1 AND TWIST2 INDUCE HUMAN MACROPHAGE MEMORY UPON CHRONIC INNATE RECEPTOR TREATMENT BY HDAC-MEDIATED DEACETYLATION OF CYTOKINE PROMOTERS. INTESTINAL TISSUES ARE CONTINUOUSLY EXPOSED TO MICROBIAL PRODUCTS THAT STIMULATE PATTERN-RECOGNITION RECEPTORS (PRRS). ONGOING PRR STIMULATION CAN CONFER EPIGENETIC CHANGES IN MACROPHAGES, WHICH CAN THEN REGULATE SUBSEQUENT IMMUNE OUTCOMES AND ADAPTATION TO THE LOCAL ENVIRONMENT. MECHANISMS LEADING TO THESE CHANGES ARE INCOMPLETELY UNDERSTOOD. WE FOUND THAT SHORT-TERM STIMULATION OF THE PRR NOD2 IN PRIMARY HUMAN MONOCYTE-DERIVED MACROPHAGES RESULTED IN INCREASED H3 AND H4 ACETYLATION OF CYTOKINE PROMOTERS, CONSISTENT WITH THE INCREASED CYTOKINE SECRETION OBSERVED. HOWEVER, WITH PROLONGED NOD2 STIMULATION, BOTH THE ACETYLATION AND CYTOKINE SECRETION WERE DRAMATICALLY DECREASED. CHRONIC NOD2 STIMULATION UPREGULATED THE TRANSCRIPTION FACTORS TWIST1 AND TWIST2, WHICH BOUND TO THE PROMOTERS OF THE HISTONE DEACETYLASES HDAC1 AND HDAC3 AND INDUCED HDAC1 AND HDAC3 EXPRESSION. HDAC1 AND HDAC3 THEN MEDIATED HISTONE DEACETYLATION AT CYTOKINE PROMOTERS AND, IN TURN, CYTOKINE DOWNREGULATION UNDER THESE CONDITIONS. SIMILAR REGULATION WAS OBSERVED UPON CHRONIC STIMULATION OF MULTIPLE PRRS. CONSISTENT WITH THE CHRONIC MICROBIAL EXPOSURE IN THE INTESTINAL ENVIRONMENT, TWIST1, TWIST2, HDAC1, AND HDAC3 WERE UPREGULATED IN HUMAN INTESTINAL RELATIVE TO PERIPHERAL MACROPHAGES. IMPORTANTLY, COMPLEMENTING HDAC1 AND HDAC3 IN TWIST1/TWIST2-DEFICIENT MONOCYTE-DERIVED MACROPHAGES RESTORED THE REDUCED HISTONE ACETYLATION ON CYTOKINE PROMOTERS AND THE DECREASED CYTOKINE SECRETION WITH CHRONIC NOD2 STIMULATION. TAKEN TOGETHER, WE IDENTIFY MECHANISMS WHEREIN TWIST1 AND TWIST2 PROMOTE CHROMATIN MODIFICATIONS, RESULTING IN MACROPHAGE INSTRUCTION AND ADAPTATION TO CONDITIONS IN THE INTESTINAL MICROENVIRONMENT. 2019 2 6891 20 [SIGNAL RECEPTORS OF CONGENITAL IMMUNITY: A NEW MOLECULAR TARGET FOR DIAGNOSTICS AND TREATMENT OF INFLAMMATORY DISEASES]. THE DISCOVERY OF SIGNAL RECEPTORS OF CONGENITAL IMMUNITY (SIGNAL PRR) NOT ONLY PROVIDED A NOVEL VIEW OF BASIC ASPECTS OF PATHOGENESIS OF CHRONIC INFLAMMATORY DISEASES BUT ALSO CREATED A BASIS FOR THE DEVELOPMENT OF ADDITIONAL DIAGNOSTIC CRITERIA FOR THESE PATHOLOGIES AND NEW PHARMACEUTICALS FOR THEIR TREATMENT. REDUCED EXPRESSION AND FUNCTION OF PRR DUE TO MUTATIONS/POLYMORPHISMS OR EPIGENETIC DISTURBANCES OF REGULATION CAN BE REGARDED AS IMMUNODEFICIENT CONDITIONS MANIFEST AS SEVERE INFECTIOUS INFLAMMATORY DISEASES. IN CONTRAST, EXCESSIVE EXPRESSION AND ACTIVATION OF PRR AS A RULE LEADS TO CHRONIC AUTOINFLAMMATORY, AUTOIMMUNE, AND ATOPIC DISEASES INVOLVING ADAPTIVE IMMUNITY AND AGGRESSION AGAINST OWN TISSUES AND CELLS. ASSESSMENT OF CERTAIN MUTATIONS IN PRR GENES, THEIR EXPRESSION AND ACTIVATION PROVIDES A POWERFUL TOOL FOR IN-DEPTH DIAGNOSTICS OF INFLAMMATORY DISEASES. SIMULTANEOUSLY, NEW LINES OF IMMUNOSTIMULATING AND ANTI-INFLAMMATORY THERAPY ARE DEVELOPED BASED ON THE KNOWLEDGE OF MOLECULAR PHYSIOLOGY OF PRR WITH THE USE OF SYNTHETIC AGONISTS AND ANTAGONISTS OF SIGNAL PRR. 2011 3 5623 19 SECOND-HAND SMOKE AND HUMAN LUNG CANCER. SINCE THE EARLY 1980S, THERE HAS BEEN GROWING CONCERN ABOUT POTENTIAL HEALTH CONSEQUENCES OF EXPOSURE TO SECOND-HAND SMOKE (SHS). DESPITE SHS BEING ESTABLISHED AS A RISK FACTOR FOR LUNG CANCER DEVELOPMENT, THE ESTIMATED RISK HAS REMAINED SMALL YET SOMEHOW DEBATABLE. HUMAN EXPOSURE TO SHS IS COMPLICATED BECAUSE OF TEMPORAL VARIABILITIES IN SOURCE, COMPOSITION, AND CONCENTRATION OF SHS. THE TEMPORALITY OF EXPOSURE TO SHS IS IMPORTANT FOR HUMAN LUNG CARCINOGENESIS WITH A LATENCY OF MANY YEARS. TO EXPLORE THE CAUSAL EFFECT OF SHS IN LUNG CARCINOGENESIS, EXPOSURE ASSESSMENTS SHOULD ESTIMATE CHRONIC EXPOSURE TO SHS ON AN INDIVIDUAL BASIS. HOWEVER, CONVENTIONAL EXPOSURE ASSESSMENT FOR SHS RELIES ON ONE-OFF OR SHORT-TERM MEASUREMENTS OF SHS INDICES. A MORE RELIABLE APPROACH WOULD BE TO USE BIOLOGICAL MARKERS THAT ARE SPECIFIC FOR SHS EXPOSURE AND PERTINENT TO LUNG CANCER. THIS APPROACH REQUIRES AN UNDERSTANDING OF THE UNDERLYING MECHANISMS THROUGH WHICH SHS COULD CONTRIBUTE TO LUNG CARCINOGENESIS. THIS REVIEW IS A SYNOPSIS OF RESEARCH ON SHS AND LUNG CANCER, WITH SPECIAL FOCUS ON HYPOTHETICAL MODES OF ACTION OF SHS FOR CARCINOGENESIS, INCLUDING GENOTOXIC AND EPIGENETIC EFFECTS. 2008 4 1667 29 DOWNREGULATION OF PCAF BY MIR-181A/B PROVIDES FEEDBACK REGULATION TO TNF-ALPHA-INDUCED TRANSCRIPTION OF PROINFLAMMATORY GENES IN LIVER EPITHELIAL CELLS. ABERRANT CELLULAR RESPONSES TO PROINFLAMMATORY CYTOKINES, SUCH AS TNF-ALPHA, ARE PATHOGENIC FEATURES IN MOST CHRONIC INFLAMMATORY DISEASES. A VARIETY OF EXTRACELLULAR AND INTRACELLULAR FEEDBACK PATHWAYS HAS EVOLVED TO PREVENT AN INAPPROPRIATE CELLULAR REACTION TO THESE PROINFLAMMATORY CYTOKINES. IN THIS STUDY, WE REPORT THAT TNF-ALPHA TREATMENT OF HUMAN AND MOUSE CHOLANGIOCYTES AND HEPATOCYTES DOWNREGULATED EXPRESSION OF P300/CBP-ASSOCIATED FACTOR (PCAF), A COACTIVATOR AND AN ACETYLTRANSFERASE THAT PROMOTES HISTONE ACETYLATION AND GENE TRANSCRIPTION. OF THESE UPREGULATED MICRORNAS IN TNF-ALPHA-TREATED CELLS, MIR-181A/B (MIR-181A AND MIR-181B) SUPPRESSED TRANSLATION OF PCAF MRNA. FUNCTIONAL MANIPULATION OF MIR-181A/B CAUSED RECIPROCAL ALTERATIONS IN PCAF PROTEIN EXPRESSION IN CULTURED CHOLANGIOCYTES AND HEPATOCYTES. INHIBITION OF MIR-181A/B FUNCTION WITH ANTI-MIRS BLOCKED TNF-ALPHA-INDUCED SUPPRESSION OF PCAF EXPRESSION. PROMOTER RECRUITMENT OF PCAF WAS SHOWN TO BE ASSOCIATED WITH TNF-ALPHA-INDUCED TRANSCRIPTION OF INFLAMMATORY GENES. INTRIGUINGLY, PRETREATMENT OF CELLS WITH TNF-ALPHA INHIBITED TRANSCRIPTION OF INFLAMMATORY GENES IN RESPONSE TO SUBSEQUENT TNF-ALPHA STIMULATION. OVEREXPRESSION OF PCAF OR INHIBITION OF MIR-181A/B FUNCTION WITH ANTI-MIRS ATTENUATED THE INHIBITORY EFFECTS OF TNF-ALPHA PRETREATMENT ON EPITHELIAL INFLAMMATORY RESPONSE TO SUBSEQUENT TNF-ALPHA STIMULATION. DOWNREGULATION OF PCAF AND THE INHIBITORY EFFECTS OF TNF-ALPHA PRETREATMENT ON LIVER EPITHELIAL INFLAMMATORY RESPONSE WERE FURTHER CONFIRMED IN A MOUSE MODEL OF TNF-ALPHA I.P. INJECTION. THESE DATA SUGGEST THAT PCAF IS A TARGET FOR MIR-181A/B, AND DOWNREGULATION OF PCAF BY TNF-ALPHA PROVIDES NEGATIVE FEEDBACK REGULATION TO INFLAMMATORY REACTIONS IN LIVER EPITHELIAL CELLS, A PROCESS THAT MAY BE RELEVANT TO THE EPIGENETIC FINE-TUNING OF EPITHELIAL INFLAMMATORY PROCESSES IN GENERAL. 2012 5 5479 23 RESVERATROL ATTENUATES CIGARETTE SMOKE EXTRACT INDUCED CELLULAR SENESCENCE IN HUMAN AIRWAY EPITHELIAL CELLS BY REGULATING THE MIR-34A/SIRT1/NF-KAPPAB PATHWAY. CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) IS CHARACTERIZED BY ACCELERATED LUNG AGING. SMOKING IS THE CRITICAL RISK FACTOR FOR COPD. CELLULAR SENESCENCE OF AIRWAY EPITHELIAL CELLS IS THE CYTOLOGICAL BASIS OF ACCELERATED LUNG AGING IN COPD, AND THE REGULATION OF MICRORNAS (MIRNAS) IS THE CENTRAL EPIGENETIC MECHANISM OF CELLULAR SENESCENCE. RESVERATROL (RES) IS A POLYPHENOL WITH ANTI-AGING PROPERTIES. THIS STUDY INVESTIGATED WHETHER RES ATTENUATES CIGARETTE SMOKE EXTRACT (CSE)-INDUCED CELLULAR SENESCENCE IN HUMAN AIRWAY EPITHELIAL CELLS (BEAS-2B) THROUGH THE MIR-34A/SIRT1/NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) PATHWAY. BEAS-2B CELLS WERE TREATED WITH RES, CSE AND TRANSFECTED WITH MIR-34A-5P MIMICS. CELLULAR SENESCENCE WAS EVALUATED BY SENESCENCE -RELATED BETA-GALACTOSIDASE (SA-BETA-GAL) STAINING AND EXPRESSION OF SENESCENCE-RELATED GENES (P16, P21, AND P53). THE EXPRESSIONS OF MIR-34A-5P, SIRT1, AND NF-KAPPAB P65 WERE EXAMINED USING QUANTITATIVE REAL TIME POLYMERASE CHAIN REACTION AND WESTERN BLOTTING. THE SENESCENCE-ASSOCIATED SECRETORY PHENOTYPE (SASP) CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) WERE ASSESSED BY ENZYME-LINKED IMMUNOSORBENT ASSAY. THE BINDING BETWEEN MIR-34A-5P AND SIRT1 WAS CONFIRMED BY DUAL-LUCIFERASE REPORTER ASSAY. THE RESULTS SHOWED THAT CSE DOSE-DEPENDENTLY DECREASED CELL VIABILITY AND ELEVATED CELLULAR SENESCENCE, CHARACTERIZED BY INCREASED SA-BETA-GAL STAINING AND SENESCENCE-RELATED GENE EXPRESSIONS (P16, P21, AND P53). FURTHER, CSE DOSE-DEPENDENTLY INCREASED THE EXPRESSION OF MIR-34A-5P AND SASP CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) IN BEAS-2B CELLS. PRETREATMENT WITH RES INHIBITED CSE-INDUCED CELLULAR SENESCENCE AND SECRETION OF SASP CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) IN A DOSE-DEPENDENT MANNER. MOREOVER, RES REVERSED THE CSE-INDUCED DOWN-REGULATION OF SIRT1 AND UP-REGULATION OF MIR-34A-5P AND NF-KAPPAB P65. SIRT1 IS A TARGET OF MIR-34A-5P. OVEREXPRESSION OF MIR-34A-5P VIA TRANSFECTION WITH MIR-34A-5P MIMIC IN BEAS-2B CELLS ATTENUATED THE INHIBITORY EFFECT OF RES ON CELLULAR SENESCENCE, ACCOMPANIED BY REVERSING THE EXPRESSION OF SIRT1 AND NF-KAPPAB P65. IN CONCLUSION, RES ATTENUATED CSE-INDUCED CELLULAR SENESCENCE IN BEAS-2B CELLS BY REGULATING THE MIR-34A/SIRT1/NF-KAPPAB PATHWAY, WHICH MAY PROVIDE A NEW APPROACH FOR COPD TREATMENT. 2022 6 6158 24 THE GENETICS AND EPIGENETICS OF ATOPIC DERMATITIS-FILAGGRIN AND OTHER POLYMORPHISMS. ATOPIC DERMATITIS (AD) IS A CHRONIC INFLAMMATORY SKIN DISEASE CAUSED BY A COMBINATION OF GENETIC AND ENVIRONMENTAL FACTORS. GENETIC EVIDENCES DEPICT A COMPLEX NETWORK COMPRISING BY EPIDERMAL BARRIER DYSFUNCTIONS AND DYSREGULATION OF INNATE AND ADAPTIVE IMMUNITY IN THE PATHOGENESIS OF AD. MUTATIONS IN THE HUMAN FILAGGRIN GENE (FLG) ARE THE MOST SIGNIFICANT AND WELL-REPLICATED GENETIC MUTATION ASSOCIATED WITH AD, AND OTHER MUTATIONS ASSOCIATED WITH EPIDERMAL BARRIERS SUCH AS SPINK5, FLG-2, SPRR3, AND CLDN1 HAVE ALL BEEN LINKED TO AD. GENE VARIANTS MAY ALSO CONTRIBUTE TO THE ABNORMAL INNATE AND ADAPTIVE RESPONSES FOUND IN AD, INCLUDING MUTATIONS IN PRRS AND AMPS, TSLP AND TSLPR, IL-1 FAMILY CYTOKINES AND RECEPTORS GENES, VITAMIN D PATHWAY GENES, FCER1A, AND TH2 AND OTHER CYTOKINES GENES. GWAS AND IMMUNOCHIP ANALYSIS HAVE IDENTIFIED A TOTAL OF 19 SUSCEPTIBILITY LOCI FOR AD. CANDIDATE GENES AT THESE SUSCEPTIBILITY LOCI IDENTIFIED BY GWAS AND IMMUNOCHIP ANALYSIS ALSO SUGGEST ROLES FOR EPIDERMAL BARRIER FUNCTIONS, INNATE AND ADAPTIVE IMMUNITY, INTERLEUKIN-1 FAMILY SIGNALING, REGULATORY T CELLS, THE VITAMIN D PATHWAY, AND THE NERVE GROWTH FACTOR PATHWAY IN THE PATHOGENESIS OF AD. INCREASING EVIDENCES SHOW THE MODERN LIFESTYLE (I.E., THE HYGIENE HYPOTHESIS, WESTERN DIET) AND OTHER ENVIRONMENTAL FACTORS SUCH AS POLLUTION AND ENVIRONMENTAL TOBACCO SMOKE (ETS) LEAD TO THE INCREASING PREVALENCE OF AD WITH THE DEVELOPMENT OF INDUSTRIALIZATION. EPIGENETIC ALTERATIONS IN RESPONSE TO THESE ENVIRONMENTAL FACTORS, INCLUDING DNA METHYLATION AND MICRORNA RELATED TO IMMUNE SYSTEM AND SKIN BARRIERS, HAVE BEEN FOUND TO CONTRIBUTE TO THE PATHOGENESIS OF AD. GENETIC VARIANTS AND EPIGENETIC ALTERATION MIGHT BE THE KEY TOOLS FOR THE MOLECULAR TAXONOMY OF AD AND PROVIDE THE BACKGROUND FOR THE PERSONALIZED MANAGEMENT. 2016 7 5227 19 PRMT6 MEDIATES INFLAMMATION VIA ACTIVATION OF THE NF-KAPPAB/P65 PATHWAY ON A CIGARETTE SMOKE EXTRACT-INDUCED MURINE EMPHYSEMA MODEL. INTRODUCTION: SMOKE-DRIVEN LUNG INFLAMMATION IS CONSIDERED TO BE THE MAJOR PATHOPHYSIOLOGY MECHANISM OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD)/EMPHYSEMA. PROTEIN ARGININE METHYLTRANSFERASE 6 (PRMT6) IS A KEY EPIGENETIC ENZYME, WHICH IS RELATED TO PROTECTING THE TRI-METHYLATION OF H3K4 (H3K4ME3). WE HYPOTHESIZED THAT PTMT6 PROTECTS LUNG INFLAMMATION THROUGH THE NUCLEAR FACTOR KAPPA B (NF-KAPPAB) PATHWAY. METHODS: MICE WERE INJECTED WITH CIGARETTE SMOKE EXTRACT (CSE) OR PBS TO ESTABLISH A MICE MODEL, INTRATRACHEALLY INSTILLED WITH OVEREXPRESSED PRMT6 OR NEGATIVE CONTROL VECTOR. MORPHOMETRY OF LUNG SLIDES AND LUNG FUNCTION WERE MEASURED. WE DETERMINED THE PROTEIN EXPRESSION OF PRMT6 AND ITS RELATED HISTONE TARGETS, THE ACTIVATION OF NF-KAPPAB PATHWAY, THE LEVEL OF TUMOR NECROSIS FACTOR ALPHA (TNFALPHA) AND INTERLEUKIN-1BETA (IL-1BETA). RESULTS: AFTER PRMT6 OVEREXPRESSION, THE MORPHOMETRY INDEXES AND LUNG FUNCTION WERE IMPROVED. ALSO, THE EXPRESSION OF H3K4ME3 WAS DECREASED. OVEREXPRESSED PRMT6 COULD SUPPRESS CSE-INDUCED NF-KAPPAB ACTIVATION AND PRO-INFLAMMATION GENES EXPRESSION. CONCLUSIONS: THE OVEREXPRESSED PRMT6 COULD SERVE AS AN INFLAMMATION INHIBITOR, POTENTIALLY THROUGH BLOCKING THE NF-KAPPAB/P65 PATHWAY IN THE MURINE EMPHYSEMA MODEL. 2020 8 5418 33 REGULATION OF DNA METHYLATION SIGNATURES ON NF-KAPPAB AND STAT3 PATHWAY GENES AND TET ACTIVITY IN CIGARETTE SMOKE EXTRACT-CHALLENGED CELLS/COPD EXACERBATION MODEL IN VITRO. BACKGROUND: CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) IS A GLOBAL HEALTH PROBLEM. CURRENTLY, THERE IS A LACK OF KNOWLEDGE ABOUT THE PATHOBIOLOGY OF THIS DISEASE AND AVAILABLE THERAPIES ARE INEFFECTIVE. CIGARETTE SMOKING IS THE LEADING CAUSE OF COPD; HOWEVER, NOT ALL SMOKERS DEVELOP COPD. EXACERBATIONS OF COPD CAUSED BY MICROBES ARE COMMON AND DETRIMENTAL. APPROXIMATELY 20-50% OF PATIENT EXACERBATIONS ARE CAUSED BY BACTERIAL COLONIZATION IN THE LOWER AIRWAYS. IT IS GENERALLY ACCEPTED THAT EPIGENETIC MECHANISMS, ESPECIALLY DNA METHYLATION, PLAY AN IMPORTANT ROLE DURING PROGRESSION OF COPD. THUS, WE HYPOTHESIZED THAT DNA METHYLATION PATTERNS VARY SIGNIFICANTLY FOLLOWING SMOKE EXPOSURE AND DURING EXACERBATIONS CAUSED BY BACTERIAL INFECTIONS. TO TEST OUR HYPOTHESIS, WE USED AN IN VITRO STUDY MODEL THAT MIMICS COPD EXACERBATIONS AND PERFORMED EXTENSIVE STUDIES TO UNDERSTAND THE ROLE OF CPG PROMOTER METHYLATION OF NF-KAPPAB AND STAT3-MEDIATED PATHWAY GENES. BOTH NF-KAPPAB AND STAT3 TRANSCRIPTION FACTORS PLAY CRITICAL ROLES IN ORCHESTRATING INFLAMMATORY RESPONSES DURING CIGARETTE SMOKE EXPOSURE. IN BRIEF, HUMAN LUNG ADENOCARCINOMA CELLS WITH TYPE II ALVEOLAR EPITHELIUM CHARACTERISTICS (A549) WERE CHALLENGED WITH CIGARETTE SMOKE EXTRACT (CSE) OR DMSO (CONTROL) FOLLOWED BY A 3-H CHALLENGE WITH BACTERIAL LIPOPOLYSACCHARIDE (LPS; FROM PSEUDOMONAS AERUGINOSA) PRIOR TO THE TERMINATION OF CSE EXPOSURE (COPD EXACERBATION GROUP). THE PRODUCTION OF CYTOKINES/CHEMOKINES, REGULATION OF TRANSCRIPTION FACTORS, AND DNA METHYLATION OF SPECIFIC GENES WERE THEN ASSESSED. WE ALSO STUDIED CHANGES IN THE EXPRESSION AND ACTIVITY OF TEN-ELEVEN TRANSLOCASES (TETS), THE ENZYMES RESPONSIBLE FOR DNA DEMETHYLATION, AND ASSESSED THEIR ROLE IN REGULATING DNA METHYLATION IN THE CSE-CHALLENGED GROUP. RESULTS: THERE WAS A SIGNIFICANT INCREASE IN THE RELEASE OF CYTOKINES/CHEMOKINES (IL-8, MCP-1, IL-6 AND CCL5) IN THE COPD EXACERBATION GROUP AS COMPARED TO THE CONTROL GROUP. HYPOMETHYLATION OF NF-KAPPAB-MEDIATED PATHWAY GENES CORRELATED WITH THEIR INDUCTION IN OUR COPD EXACERBATION STUDY MODEL. FURTHER, WE OBSERVED AN IMPORTANT ROLE OF TET1/2 IN REGULATING THE DNA METHYLATION OF NF-KAPPAB, STAT3, IKK, AND NIK GENES AND CYTOKINE/CHEMOKINE PRODUCTION BY A549 CELLS DURING CSE CHALLENGE. CONCLUSIONS: STUDIES TO FURTHER DEFINE THE ROLE OF TETS IN CSE-MEDIATED EPIGENETIC REGULATION MAY LEAD TO THE DEVELOPMENT OF BETTER AND MORE EFFECTIVE THERAPEUTIC INTERVENTION STRATEGIES FOR COPD. 2020 9 6600 24 TWIST2 DEMONSTRATES DIFFERENTIAL METHYLATION IN IMMUNOGLOBULIN VARIABLE HEAVY CHAIN MUTATED AND UNMUTATED CHRONIC LYMPHOCYTIC LEUKEMIA. PURPOSE: CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS A CLINICALLY HETEROGENEOUS DISEASE FOR WHICH NATURAL HISTORY CAN BE PREDICTED BASED ON THE PRESENCE OR ABSENCE OF IMMUNOGLOBULIN (IG) VARIABLE HEAVY CHAIN (V(H)) GENE MUTATIONS. HEREIN WE REPORT SELECTIVE EPIGENETIC SILENCING OF THE TRANSCRIPTION FACTOR TWIST2 (DERMO1) IN IG V(H) MUTATED CLL AND DESCRIBE A SEMIQUANTITATIVE ASSAY TO STUDY PROMOTER METHYLATION OF THIS GENE IN PRIMARY TUMOR CELLS. MATERIALS AND METHODS: TWIST2 PROMOTER METHYLATION WAS IDENTIFIED BY RESTRICTION LANDMARK GENOME SCANNING. SOUTHERN BLOT (SB), BISULFITE SEQUENCING, AND COMBINED BISULFITE RESTRICTION ANALYSIS (COBRA), AND QUANTITATIVE SB-COBRA WAS PERFORMED TO STUDY METHYLATION OF THE TWIST2 PROMOTER. REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION ASSAYS WERE USED TO STUDY TWIST2 EXPRESSION IN CLL CELLS. RESULTS: FOLLOWING IDENTIFICATION AND CONFIRMATION OF TWIST2 METHYLATION IN CLL PATIENTS, WE DEMONSTRATED THAT EXPRESSION OF THIS TRANSCRIPTION FACTOR IS RELATED TO THE DEGREE OF PROMOTER METHYLATION. EXPRESSION OF TWIST2 IN A CLL CELL LINE IN WHICH THE PROMOTER IS METHYLATED WAS INCREASED FOLLOWING DECITABINE TREATMENT. WE NEXT STUDIED 53 PATIENTS BY COBRA AND DEMONSTRATED THAT 72% OF PATIENT SAMPLES WITH MUTATED IG V(H) SHOW TWIST2 METHYLATION, WHILE ONLY 16% OF PATIENT SAMPLES WITH UNMUTATED IG V(H) WERE METHYLATED (P < .001). IN A SUBSET OF PATIENTS, METHYLATION OF TWIST2 CORRELATED WITH MRNA EXPRESSION. CONCLUSION: TWIST2 IS DIFFERENTIALLY METHYLATED IN CLL CELLS RELATIVE TO IG V(H) MUTATIONAL STATUS AND CAN BE QUANTITATIVELY MONITORED BY SB-COBRA. BASED ON THE KNOWN ROLE OF TWIST2 IN SILENCING P53 FUNCTION IN OTHER MALIGNANCIES, FUTURE STUDIES SHOULD FOCUS ON THE ROLE OF TWIST2 IN CLL AND RELATED LYMPHOPROLIFERATIVE DISEASES. 2005 10 3940 27 LNCRNA DLEU2 REGULATES SIRTUINS AND MITOCHONDRIAL RESPIRATORY CHAIN COMPLEX IV: A NOVEL PATHWAY IN OBESITY AND OFFSPRING'S HEALTH. BACKGROUND: LONG NON-CODING RNAS (LNCRNAS) HAVE EMERGED AS A RAPIDLY EXPANDING AREA OF INTEREST IN CHRONIC DISEASES. THEY ARE MOSTLY UNKNOWN FOR ROLES IN METABOLIC REGULATION. SIRTUINS, AN EPIGENETIC MODULATOR CLASS, REGULATE METABOLIC PATHWAYS. HOWEVER, HOW SIRTUINS ARE REGULATED VIA LNCRNA IS UNKNOWN. WE HYPOTHESIZED THAT A HIGH-FAT HIGH-FRUCTOSE DIET (HFD-HF) DURING PREGNANCY WOULD INCREASE THE RISK FOR OBESITY VIA LNCRNA-SIRTUIN PATHWAYS. METHODS: FEMALE C57BL/6 MICE (F0) WERE FED EITHER CHOW DIET (CD) OR HFD-HF FOR 6 WEEKS TILL BIRTH. THE PUPS (F1) WERE FED EITHER CD OR HFD-HF FOR 20 WEEKS. EXPRESSION OF DLEU2, SIRTUINS, MITOCHONDRIAL RESPIRATORY COMPLEXES, AND OXIDATIVE STRESS WERE INVESTIGATED IN THE F1 LIVERS. FASTING BLOOD GLUCOSE, INSULIN SENSITIVITY, GLUCOSE TOLERANCE, BODY AND TISSUES WEIGHT WERE MEASURED. A MECHANISTIC INTERACTION WAS THEN CARRIED OUT USING A DLEU2 KNOCKDOWN EXPERIMENT IN THE HEPG2 CELL. RESULTS: DLEU2 AND SIRTUINS WERE BOTH SIGNIFICANTLY DECREASED IN THE LIVERS OF HFD-HF FED MALE F1 WHOSE MOTHERS WERE EITHER FED CD OR HFD-HF DURING REPRODUCTIVE AND PREGNANCY WINDOWS. CONFIRMING THIS CONNECTION, UPON SILENCING DLEU2, TRANSCRIPTION LEVELS OF SIRT1 THROUGH 6 AND TRANSLATIONAL LEVELS OF SIRT1, 3, 5, AND 6 WERE SIGNIFICANTLY DOWNREGULATED. KNOCKDOWN OF DLEU2 SIGNIFICANTLY DECREASED THE PROTEIN LEVEL OF CYTOCHROME-C OXIDASE (COMPLEX IV, MTCO1) WITHOUT ALTERING OTHER MITOCHONDRIAL COMPLEXES, DECREASED MITOCHONDRIAL MEMBRANE POTENTIAL, DECREASED ATP, AND INCREASED REACTIVE OXYGEN SPECIES. INTERESTINGLY, IN F1 LIVERS, THE PROTEIN LEVEL OF MTCO1 WAS ALSO SIGNIFICANTLY DECREASED UNDER AN HFD-HF DIET OR EVEN UNDER CHOW DIET IF THE MOTHER WAS EXPOSED TO HFD-HF. CONCLUSION: OUR FINDINGS REVEAL FOR THE FIRST TIME THAT ONE LNCRNA CAN REGULATE SIRTUINS AND A SPECIFIC MITOCHONDRIAL COMPLEX. FURTHERMORE, DIET OR MATERNAL DIET CAN MODULATE DLEU2 AND ITS DOWNSTREAM REGULATORS IN OFFSPRING, SUGGESTING A POTENTIAL ROLE OF DLEU2 IN METABOLIC DISORDERS OVER ONE OR MORE GENERATIONS. 2022 11 3983 26 LONG-TERM EXPOSURE TO CIGARETTE SMOKE EXTRACT INDUCES HYPOMETHYLATION AT THE RUNX3 AND IGF2-H19 LOCI IN IMMORTALIZED HUMAN UROTHELIAL CELLS. CIGARETTE SMOKING IS THE SINGLE MOST IMPORTANT EPIDEMIOLOGICAL RISK FACTOR FOR BLADDER CANCER BUT IT IS NOT KNOWN WHETHER EXPOSURE OF UROTHELIAL CELLS TO THE SYSTEMIC SOLUBLE CONTENTS OF CIGARETTE SMOKE IS DIRECTLY CAUSATIVE TO BLADDER CANCER AND THE ASSOCIATED EPIGENETIC CHANGES SUCH AS TUMOR SUPPRESSOR GENE HYPERMETHYLATION. WE UNDERTOOK THIS STUDY TO INVESTIGATE IF LONG-TERM TREATMENT OF HUMAN UROTHELIAL CELLS WITH CIGARETTE SMOKE EXTRACT (CSE) RESULTS IN TUMOR SUPPRESSOR GENE HYPERMETHYLATION, A PHENOTYPE THAT WAS PREVIOUSLY ASSOCIATED WITH LONG-TERM CONSTANT CSE TREATMENT OF AIRWAY EPITHELIAL CELLS. WE CHRONICALLY TREATED AN IMMORTALIZED HUMAN UROTHELIAL CELL LINE UROTSA WITH CSE USING A CYCLIC DAILY REGIMEN BUT THE CELLS WERE CULTURED IN CSE-FREE MEDIUM BETWEEN DAILY TREATMENTS. BISULFITE SEQUENCING AND REAL-TIME PCR ARRAY-BASED METHYLATION PROFILING WERE EMPLOYED TO EVALUATE METHYLATION CHANGES AT TUMOR SUPPRESSOR GENE LOCI IN THE CHRONICALLY CSE-TREATED CELLS VERSUS THE PASSAGE-MATCHED UNTREATED CONTROL CELLS. THE RUNX3 TUMOR SUPPRESSOR GENE PROMOTER WAS HYPOMETHYLATED WITH A SIGNIFICANT INCREASE IN PROPORTION OF THE COMPLETELY UNMETHYLATED HAPLOTYPE AFTER THE LONG-TERM CSE TREATMENT; WHEREAS RUNX3 PROMOTER HYPERMETHYLATION WAS PREVIOUSLY REPORTED FOR BLADDER CANCERS OF SMOKERS. HYPOMETHYLATION INDUCED BY THE LONG-TERM CSE TREATMENT WAS ALSO OBSERVED FOR THE IGF2-H19 LOCUS. THE METHYLATION STATUS AT THE PRSS8/PROSTASIN AND 16 ADDITIONAL LOCI HOWEVER, WAS UNAFFECTED BY THE CHRONIC CSE TREATMENT. TRANSIENT CSE TREATMENT OVER 1 DAILY REGIMEN RESULTED IN TRANSCRIPTIONAL DOWN-REGULATION OF RUNX3 AND H19, BUT ONLY THE H19 TRANSCRIPTION WAS DOWN-REGULATED IN THE CHRONICALLY CSE-TREATED UROTHELIAL CELLS. TRANSCRIPTION OF A KEY ENZYME IN ONE-CARBON METABOLISM, DIHYDROFOLATE REDUCTASE (DHFR) WAS GREATLY REDUCED BY THE LONG-TERM CSE TREATMENT, POTENTIALLY SERVING AS A MECHANISM FOR THE HYPOMETHYLATION PHENOTYPE VIA A REDUCED SUPPLY OF METHYL DONOR. IN CONCLUSION, CHRONIC CYCLIC CSE TREATMENT OF UROTHELIAL CELLS INDUCED HYPOMETHYLATION RATHER THAN HYPERMETHYLATION AT SPECIFIC LOCI. 2013 12 2910 29 GENE EXPRESSION PROFILING OF EPIGENETIC CHROMATIN MODIFICATION ENZYMES AND HISTONE MARKS BY CIGARETTE SMOKE: IMPLICATIONS FOR COPD AND LUNG CANCER. CHROMATIN-MODIFYING ENZYMES MEDIATE DNA METHYLATION AND HISTONE MODIFICATIONS ON RECRUITMENT TO SPECIFIC TARGET GENE LOCI IN RESPONSE TO VARIOUS STIMULI. THE KEY ENZYMES THAT REGULATE CHROMATIN ACCESSIBILITY FOR MAINTENANCE OF MODIFICATIONS IN DNA AND HISTONES, AND FOR MODULATION OF GENE EXPRESSION PATTERNS IN RESPONSE TO CIGARETTE SMOKE (CS), ARE NOT KNOWN. WE HYPOTHESIZE THAT CS EXPOSURE ALTERS THE GENE EXPRESSION PATTERNS OF CHROMATIN-MODIFYING ENZYMES, WHICH THEN AFFECTS MULTIPLE DOWNSTREAM PATHWAYS INVOLVED IN THE RESPONSE TO CS. WE HAVE, THEREFORE, ANALYZED CHROMATIN-MODIFYING ENZYME PROFILES AND VALIDATED BY QUANTITATIVE REAL-TIME PCR (QPCR). WE ALSO PERFORMED IMMUNOBLOT ANALYSIS OF TARGETED HISTONE MARKS IN C57BL/6J MICE EXPOSED TO ACUTE AND SUBCHRONIC CS, AND OF LUNGS FROM NONSMOKERS, SMOKERS, AND PATIENTS WITH CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD). WE FOUND A SIGNIFICANT INCREASE IN EXPRESSION OF SEVERAL CHROMATIN MODIFICATION ENZYMES, INCLUDING DNA METHYLTRANSFERASES, HISTONE ACETYLTRANSFERASES, HISTONE METHYLTRANSFERASES, AND SET DOMAIN PROTEINS, HISTONE KINASES, AND UBIQUITINASES. OUR QPCR VALIDATION DATA REVEALED A SIGNIFICANT DOWNREGULATION OF DNMT1, DNMT3A, DNMT3B, HDAC2, HDAC4, HAT1, PRMT1, AND AURKB WE IDENTIFIED TARGETED CHROMATIN HISTONE MARKS (H3K56AC AND H4K12AC), WHICH ARE INDUCED BY CS. THUS CS-INDUCED GENOTOXIC STRESS DIFFERENTIALLY AFFECTS THE EXPRESSION OF EPIGENETIC MODULATORS THAT REGULATE TRANSCRIPTION OF TARGET GENES VIA DNA METHYLATION AND SITE-SPECIFIC HISTONE MODIFICATIONS. THIS MAY HAVE IMPLICATIONS IN DEVISING EPIGENETIC-BASED THERAPIES FOR COPD AND LUNG CANCER. 2016 13 2973 26 GENETIC AND EPIGENETIC STUDIES OF ATOPIC DERMATITIS. BACKGROUND: ATOPIC DERMATITIS (AD) IS A CHRONIC INFLAMMATORY DISEASE CAUSED BY THE COMPLEX INTERACTION OF GENETIC, IMMUNE AND ENVIRONMENTAL FACTORS. THERE HAVE MANY RECENT DISCOVERIES INVOLVING THE GENETIC AND EPIGENETIC STUDIES OF AD. METHODS: A RETROSPECTIVE PUBMED SEARCH WAS CARRIED OUT FROM JUNE 2009 TO JUNE 2016 USING THE TERMS "ATOPIC DERMATITIS", "ASSOCIATION", "ECZEMA", "GENE", "POLYMORPHISM", "MUTATION", "VARIANT", "GENOME WIDE ASSOCIATION STUDY", "MICROARRAY" "GENE PROFILING", "RNA SEQUENCING", "EPIGENETICS" AND "MICRORNA". A TOTAL OF 132 PUBLICATIONS IN ENGLISH WERE IDENTIFIED. RESULTS: TO ELUCIDATE THE GENETIC FACTORS FOR AD PATHOGENESIS, CANDIDATE GENE ASSOCIATION STUDIES, GENOME-WIDE ASSOCIATION STUDIES (GWAS) AND TRANSCRIPTOMIC PROFILING ASSAYS HAVE BEEN PERFORMED IN THIS PERIOD. EPIGENETIC MECHANISMS FOR AD DEVELOPMENT, INCLUDING GENOMIC DNA MODIFICATION AND MICRORNA POSTTRANSCRIPTIONAL REGULATION, HAVE BEEN EXPLORED. TO DATE, CANDIDATE GENE ASSOCIATION STUDIES INDICATE THAT FILAGGRIN (FLG) NULL GENE MUTATIONS ARE THE MOST SIGNIFICANT KNOWN RISK FACTOR FOR AD, AND GENES IN THE TYPE 2 T HELPER LYMPHOCYTE (TH2) SIGNALING PATHWAYS ARE THE SECOND REPLICATED GENETIC RISK FACTOR FOR AD. GWAS STUDIES IDENTIFIED 34 RISK LOCI FOR AD, THESE LOCI ALSO SUGGEST THAT GENES IN IMMUNE RESPONSES AND EPIDERMAL SKIN BARRIER FUNCTIONS ARE ASSOCIATED WITH AD. ADDITIONALLY, GENE PROFILING ASSAYS DEMONSTRATED AD IS ASSOCIATED WITH DECREASED GENE EXPRESSION OF EPIDERMAL DIFFERENTIATION COMPLEX GENES AND ELEVATED TH2 AND TH17 GENES. HYPOMETHYLATION OF TSLP AND FCER1G IN AD WERE REPORTED; AND MIR-155, WHICH TARGET THE IMMUNE SUPPRESSOR CTLA-4, WAS FOUND TO BE SIGNIFICANTLY OVER-EXPRESSED IN INFILTRATING T CELLS IN AD SKIN LESIONS. CONCLUSIONS: THE RESULTS SUGGEST THAT TWO MAJOR BIOLOGIC PATHWAYS ARE RESPONSIBLE FOR AD ETIOLOGY: SKIN EPITHELIAL FUNCTION AND INNATE/ADAPTIVE IMMUNE RESPONSES. THE DYSFUNCTIONAL EPIDERMAL BARRIER AND IMMUNE RESPONSES RECIPROCALLY AFFECT EACH OTHER, AND THEREBY DRIVE DEVELOPMENT OF AD. 2016 14 5760 32 SOLUBLE URIC ACID PRIMES TLR-INDUCED PROINFLAMMATORY CYTOKINE PRODUCTION BY HUMAN PRIMARY CELLS VIA INHIBITION OF IL-1RA. OBJECTIVES: THE STUDY OF THE PROINFLAMMATORY ROLE OF URIC ACID HAS FOCUSED ON THE EFFECTS OF ITS CRYSTALS OF MONOSODIUM URATE (MSU). HOWEVER, LITTLE IS KNOWN WHETHER URIC ACID ITSELF CAN DIRECTLY HAVE PROINFLAMMATORY EFFECTS. IN THIS STUDY, WE INVESTIGATE THE PRIMING EFFECTS OF URIC ACID EXPOSURE ON THE CYTOKINE PRODUCTION OF PRIMARY HUMAN CELLS UPON STIMULATION WITH GOUT-RELATED STIMULI. METHODS: PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) WERE HARVESTED FROM PATIENTS WITH GOUT AND HEALTHY VOLUNTEERS. CELLS WERE PRETREATED WITH OR WITHOUT URIC ACID IN SOLUBLE FORM FOR 24 H AND THEN STIMULATED FOR 24 H WITH TOLL-LIKE RECEPTOR (TLR)2 OR TLR4 LIGANDS IN THE PRESENCE OR ABSENCE OF MSU CRYSTALS. CYTOKINE PRODUCTION WAS MEASURED BY ELISA; MRNA LEVELS WERE ASSESSED USING QPCR. RESULTS: THE PRODUCTION OF INTERLEUKIN (IL)-1BETA AND IL-6 WAS HIGHER IN PATIENTS COMPARED WITH CONTROLS AND THIS CORRELATED WITH SERUM URATE LEVELS. PROINFLAMMATORY CYTOKINE PRODUCTION WAS SIGNIFICANTLY POTENTIATED WHEN CELLS FROM HEALTHY SUBJECTS WERE PRETREATED WITH URIC ACID. SURPRISINGLY, THIS WAS ASSOCIATED WITH A SIGNIFICANT DOWNREGULATION OF THE ANTI-INFLAMMATORY CYTOKINE IL-1 RECEPTOR ANTAGONIST (IL-1RA). THIS EFFECT WAS SPECIFIC TO STIMULATION BY URIC ACID AND WAS EXERTED AT THE LEVEL OF GENE TRANSCRIPTION. EPIGENETIC REPROGRAMMING AT THE LEVEL OF HISTONE METHYLATION BY URIC ACID WAS INVOLVED IN THIS EFFECT. CONCLUSIONS: IN THIS STUDY WE DEMONSTRATE A MECHANISM THROUGH WHICH HIGH CONCENTRATIONS OF URIC ACID (UP TO 50 MG/DL) INFLUENCE INFLAMMATORY RESPONSES BY FACILITATING IL-1BETA PRODUCTION IN PBMCS. WE SHOW THAT A MECHANISM FOR THE AMPLIFICATION OF IL-1BETA CONSISTS IN THE DOWNREGULATION OF IL-1RA AND THAT THIS EFFECT COULD BE EXERTED VIA EPIGENETIC MECHANISMS SUCH AS HISTONE METHYLATION. HYPERURICAEMIA CAUSES A SHIFT IN THE IL-1BETA/IL-1RA BALANCE PRODUCED BY PBMCS AFTER EXPOSURE TO MSU CRYSTALS AND TLR-MEDIATED STIMULI, AND THIS PHENOMENON IS LIKELY TO REINFORCE THE ENHANCED STATE OF CHRONIC INFLAMMATION. 2016 15 3767 28 INTEGRATIVE EPIGENOMIC ANALYSIS IN DIFFERENTIATED HUMAN PRIMARY BRONCHIAL EPITHELIAL CELLS EXPOSED TO CIGARETTE SMOKE. CIGARETTE SMOKE (CS) IS ONE OF THE MAJOR RISK FACTORS FOR MANY PULMONARY DISEASES, INCLUDING CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) AND LUNG CANCER. THE FIRST LINE OF DEFENSE FOR CS EXPOSURE IS THE BRONCHIAL EPITHELIAL CELLS. ELUCIDATION OF THE EPIGENETIC CHANGES DURING CS EXPOSURE IS KEY TO GAINING A MECHANISTIC UNDERSTANDING INTO HOW MATURE AND DIFFERENTIATED BRONCHIAL EPITHELIAL CELLS RESPOND TO CS. THEREFORE, WE PERFORMED EPIGENOMIC PROFILING IN CONJUNCTION WITH TRANSCRIPTIONAL PROFILING IN WELL-DIFFERENTIATED HUMAN BRONCHIAL EPITHELIAL (HBE) CELLS CULTURED IN AIR-LIQUID INTERFACE (ALI) EXPOSED TO THE VAPOR PHASE OF CS. THE GENOME-WIDE ENRICHMENT OF HISTONE 3 LYSINE 27 ACETYLATION WAS DETECTED BY CHROMATIN IMMUNOPRECIPITATION FOLLOWED BY NEXT GENERATION SEQUENCING (CHIP-SEQ) IN HBE CELLS AND SUGGESTED THE PLAUSIBLE BINDING OF SPECIFIC TRANSCRIPTION FACTORS RELATED TO CS EXPOSURE. ADDITIONALLY, INTERROGATION OF CHIP-SEQ DATA WITH GENE EXPRESSION PROFILING OF HBE CELLS AFTER CS EXPOSURE FOR DIFFERENT DURATIONS (3 HOURS, 2 DAYS, 4 DAYS) SUGGESTED THAT EARLIER EPIGENETIC CHANGES (3 HOURS AFTER CS EXPOSURE) MAY BE ASSOCIATED WITH LATER GENE EXPRESSION CHANGES INDUCED BY CS EXPOSURE (4 DAYS). THE INTEGRATION OF EPIGENETICS AND GENE EXPRESSION DATA REVEALED SIGNALING PATHWAYS RELATED TO CS-INDUCED EPIGENETIC CHANGES IN HBE CELLS THAT MAY IDENTIFY NOVEL REGULATORY PATHWAYS RELATED TO CS-INDUCED COPD. 2018 16 5417 27 REGULATION OF DNA METHYLATION IN RHEUMATOID ARTHRITIS SYNOVIOCYTES. RHEUMATOID ARTHRITIS (RA) IS A CHRONIC INFLAMMATORY DISEASE IN WHICH FIBROBLAST-LIKE SYNOVIOCYTES (FLS) EXHIBIT AN AGGRESSIVE PHENOTYPE. ALTHOUGH THE MECHANISMS RESPONSIBLE ARE NOT WELL DEFINED, EPIGENETIC DETERMINANTS SUCH AS DNA METHYLATION MIGHT CONTRIBUTE. DNA METHYLTRANSFERASES (DNMTS) ARE CRITICAL ENZYMES THAT ESTABLISH AND MAINTAIN DNA METHYLATION. WE EVALUATED WHETHER PROINFLAMMATORY CYTOKINES MIGHT CONTRIBUTE TO DIFFERENTIAL DNA METHYLATION PREVIOUSLY DESCRIBED IN RA FLS THROUGH ALTERED DNMT EXPRESSION. FLS WERE OBTAINED FROM RA AND OSTEOARTHRITIS (OA) SYNOVIUM AT THE TIME OF TOTAL JOINT REPLACEMENT. GENE EXPRESSION WAS DETERMINED BY QUANTITATIVE REAL-TIME PCR AND PROTEIN EXPRESSION BY WESTERN BLOT ANALYSIS. DNMT ACTIVITY WAS MEASURED WITH A FUNCTIONAL ASSAY, AND GLOBAL METHYLATION WAS DETERMINED BY AN IMMUNOASSAY THAT DETECTS METHYLCYTOSINE. RESTING EXPRESSION OF DNMT1, -3A, AND -3B MRNA WERE SIMILAR IN RA AND OA FLS. WESTERN BLOT SHOWED ABUNDANT DNMT1 AND DNMT3A PROTEIN. EXPOSURE TO IL-1 DECREASED DNMT1 AND DNMT3A MRNA EXPRESSION IN FLS. DOSE RESPONSES DEMONSTRATED DECREASED DNMT EXPRESSION AT CONCENTRATIONS AS LOW AS 1 PG/ML OF IL-1. DNMT MRNA LEVELS DECREASED RAPIDLY, WITH SIGNIFICANT SUPPRESSION AFTER 2-8 H OF IL-1 STIMULATION. IL-1 STIMULATION OF OA FLS DID NOT AFFECT METHYLATION OF LINE1 SITES BUT LED TO DEMETHYLATION OF A CHI3L1 LOCUS THAT IS HYPOMETHYLATED IN RA FLS. CHRONIC IL-1 STIMULATION ALSO MIMICKED THE EFFECT OF A DNMT INHIBITOR ON FLS GENE EXPRESSION. EXPOSURE TO PROINFLAMMATORY MEDIATORS REVERSIBLY ALTERS DNA METHYLATION IN FLS BY DECREASING DNMT EXPRESSION AND FUNCTION. THESE DATA SUGGEST THAT IL-1 CAN POTENTIALLY IMPRINT CELLS IN CHRONIC INFLAMMATORY DISEASES. 2013 17 4081 28 MATERNAL MICRONUTRIENT SUPPLEMENTATION SUPPRESSES T CELL CHEMOKINE RECEPTOR EXPRESSION AND FUNCTION IN F1 MICE. PRENATAL ENVIRONMENTAL EXPOSURES PLAY A CRITICAL ROLE IN DETERMINING LATE-LIFE CHRONIC DISEASE SUSCEPTIBILITY. HOWEVER, THE MECHANISMS LINKING THE IN UTERO ENVIRONMENT AND DISEASE DEVELOPMENT IN THE OFFSPRING ARE POORLY UNDERSTOOD. RECENT INVESTIGATIONS HAVE CONFIRMED A CENTRAL PATHOGENIC ROLE OF T CELL CHEMOKINE RECEPTORS, PARTICULARLY C-C CHEMOKINE RECEPTOR (CCR) 2 AND CCR5, IN CHRONIC INFLAMMATORY CONDITIONS. THIS STUDY WAS DESIGNED TO DETERMINE THE EFFECT OF A SYNTHETIC PRENATAL MICRONUTRIENT SUPPLEMENTATION (MS) DIET RICH IN METHIONINE PATHWAY METABOLITES ON THE T CELL CHEMOKINE SYSTEM IN F1 C57BL/6 MICE. FEMALE MICE WERE FED EITHER AN MS OR CONTROL DIET 3 WK PRIOR TO MATING, DURING PREGNANCY, AND LACTATION. AT 4 WK OF AGE, F1 MICE WERE KILLED FOR EXPERIMENTS OR WERE FED THE STANDARD NIH-31 DIET AND ALLOWED TO AGE. FOOD CONSUMPTION, MATERNAL WEIGHT GAIN, AND LITTER SIZE WERE SIMILAR IN DAMS FED THE CONTROL AND MS DIETS. HOWEVER, THE F1 OFFSPRING OF DAMS FED THE MS DIET WERE SMALLER IN SIZE (P < 0.001). T CELLS FROM THE MS F1 OFFSPRING HAD GLOBAL HYPERMETHYLATION COMPARED WITH CONTROL F1 OFFSPRING (P < 0.005), CORRESPONDING TO LOWER T CELL CHEMOKINE RECEPTOR EXPRESSION [CCR2 (P < 0.001), CCR5 (P < 0.001), AND C-X-C CHEMOKINE RECEPTOR 3 (P < 0.01)] AND CYTOKINE EXPRESSION [TNFALPHA (P < 0.05), IL-2 (P < 0.001), AND IL-4 (P < 0.01)]. REDUCED T CELL CHEMOKINE RECEPTOR GENE EXPRESSION IN MS F1 MICE WAS ASSOCIATED WITH DECREASED CHEMOTAXIS IN VITRO TO C-C CHEMOKINE LIGAND (CCL) 2 AND C-X-C CHEMOKINE LIGAND 10 (P < 0.01) AND IN VIVO TO CCL2 (P < 0.01). TAKEN TOGETHER, THE RESULTS SUGGEST THAT EPIGENETIC ALTERATION THROUGH PRENATAL DIET MANIPULATION REDUCES THE RESPONSE TO PROINFLAMMATORY SIGNALS IN MICE. 2012 18 2940 34 GENETIC AND EPIGENETIC ALTERATIONS IN NORMAL AND SENSITIVE COPD-DISEASED HUMAN BRONCHIAL EPITHELIAL CELLS REPEATEDLY EXPOSED TO AIR POLLUTION-DERIVED PM(2.5). EVEN THOUGH CLINICAL, EPIDEMIOLOGICAL AND TOXICOLOGICAL STUDIES HAVE PROGRESSIVELY PROVIDED A BETTER KNOWLEDGE OF THE UNDERLYING MECHANISMS BY WHICH AIR POLLUTION-DERIVED PARTICULATE MATTER (PM) EXERTS ITS HARMFUL HEALTH EFFECTS, FURTHER IN VITRO STUDIES ON RELEVANT CELL SYSTEMS ARE STILL NEEDED. HENCE, AIMING OF GETTING CLOSER TO THE HUMAN IN VIVO CONDITIONS, PRIMARY HUMAN BRONCHIAL EPITHELIAL CELLS DERIVED FROM NORMAL SUBJECTS (NHBE) OR SENSITIVE CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD)-DISEASED PATIENTS (DHBE) WERE DIFFERENTIATED AT THE AIR-LIQUID INTERFACE. THEREAFTER, THEY WERE REPEATEDLY EXPOSED TO AIR POLLUTION-DERIVED PM(2.5) TO STUDY THE OCCURRENCE OF SOME RELEVANT GENETIC AND/OR EPIGENETIC ENDPOINTS. CONCENTRATION-, EXPOSURE- AND SEASON-DEPENDENT INCREASES OF OH-B[A]P METABOLITES IN NHBE, AND TO A LESSER EXTENT, COPD-DHBE CELLS WERE REPORTED; HOWEVER, THERE WERE MORE TETRA-OH-B[A]P AND 8-OHDG DNA ADDUCTS IN COPD-DHBE CELLS. NO INCREASE IN PRIMARY DNA STRAND BREAK NOR CHROMOSOMAL ABERRATION WAS OBSERVED IN REPEATEDLY EXPOSED CELLS. TELOMERE LENGTH AND TELOMERASE ACTIVITY WERE MODIFIED IN A CONCENTRATION- AND EXPOSURE-DEPENDENT MANNER IN NHBE AND PARTICULARLY COPD-DHBE CELLS. THERE WERE A GLOBAL DNA HYPOMETHYLATION, A P16 GENE PROMOTER HYPERMETHYLATION, AND A DECREASING DNA METHYLTRANSFERASE ACTIVITY IN NHBE AND NOTABLY COPD-DHBE CELLS REPEATEDLY EXPOSED. CHANGES IN SITE-SPECIFIC METHYLATION, ACETYLATION, AND PHOSPHORYLATION OF HISTONE H3 (I.E., H3K4ME3, H3K9AC, H3K27AC, AND H3S10PH) AND RELATED ENZYME ACTIVITIES OCCURRED IN A CONCENTRATION- AND EXPOSURE-DEPENDENT MANNER IN ALL THE REPEATEDLY EXPOSED CELLS. COLLECTIVELY, THESE RESULTS HIGHLIGHTED THE KEY ROLE PLAYED BY GENETIC AND EVEN EPIGENETIC EVENTS IN NHBE AND PARTICULARLY SENSITIVE COPD-DHBE CELLS REPEATEDLY EXPOSED TO AIR POLLUTION-DERIVED PM(2.5) AND THEIR DIFFERENT RESPONSIVENESS. WHILE THESE SPECIFIC EPIGENETIC CHANGES HAVE BEEN ALREADY DESCRIBED IN COPD AND EVEN LUNG CANCER PHENOTYPES, OUR FINDINGS SUPPORTED THAT, TOGETHER WITH GENETIC EVENTS, THESE EPIGENETIC EVENTS COULD DRAMATICALLY CONTRIBUTE TO THE SHIFT FROM HEALTHY TO DISEASED PHENOTYPES FOLLOWING REPEATED EXPOSURE TO RELATIVELY LOW DOSES OF AIR POLLUTION-DERIVED PM(2.5). 2017 19 4506 25 MRTF-A MEDIATES LPS-INDUCED PRO-INFLAMMATORY TRANSCRIPTION BY INTERACTING WITH THE COMPASS COMPLEX. CHRONIC INFLAMMATION UNDERSCORES THE PATHOGENESIS OF A RANGE OF HUMAN DISEASES. LIPOPOLYSACCHARIDE (LPS) ELICITS STRONG PRO-INFLAMMATORY RESPONSES IN MACROPHAGES THROUGH THE TRANSCRIPTION FACTOR NF-KAPPAB. THE EPIGENETIC MECHANISM UNDERLYING LPS-INDUCED PRO-INFLAMMATORY TRANSCRIPTION IS NOT FULLY UNDERSTOOD. HEREIN, WE DESCRIBE A ROLE FOR MYOCARDIN-RELATED TRANSCRIPTION FACTOR A (MRTF-A, ALSO KNOWN AS MKL1) IN THIS PROCESS. MRTF-A OVEREXPRESSION ENHANCED NF-KAPPAB-DEPENDENT PRO-INFLAMMATORY TRANSCRIPTION, WHEREAS MRTF-A SILENCING INHIBITED THIS PROCESS. MRTF-A DEFICIENCY ALSO REDUCED THE SYNTHESIS OF PRO-INFLAMMATORY MEDIATORS IN A MOUSE MODEL OF COLITIS. LPS PROMOTED THE RECRUITMENT OF MRTF-A TO THE PROMOTERS OF PRO-INFLAMMATORY GENES IN AN NF-KAPPAB-DEPENDENT MANNER. RECIPROCALLY, MRTF-A INFLUENCED THE NUCLEAR ENRICHMENT AND TARGET BINDING OF NF-KAPPAB. MECHANISTICALLY, MRTF-A WAS NECESSARY FOR THE ACCUMULATION OF ACTIVE HISTONE MODIFICATIONS ON NF-KAPPAB TARGET PROMOTERS BY COMMUNICATING WITH THE HISTONE H3K4 METHYLTRANSFERASE COMPLEX (COMPASS). SILENCING OF INDIVIDUAL MEMBERS OF COMPASS, INCLUDING ASH2, WDR5 AND SET1 (ALSO KNOWN AS SETD1A), DOWNREGULATED THE PRODUCTION OF PRO-INFLAMMATORY MEDIATORS AND IMPAIRED THE NF-KAPPAB KINETICS. IN SUMMARY, OUR WORK HAS UNCOVERED A PREVIOUSLY UNKNOWN FUNCTION FOR MRTF-A AND PROVIDED INSIGHTS INTO THE RATIONALIZED DEVELOPMENT OF ANTI-INFLAMMATORY THERAPEUTIC STRATEGIES. 2014 20 4041 32 MACROPHAGE PLASTICITY IN DUCHENNE MUSCULAR DYSTROPHY: A NEXUS OF PATHOLOGICAL REMODELLING WITH THERAPEUTIC IMPLICATIONS. DUCHENNE MUSCULAR DYSTROPHY (DMD) IS CHARACTERIZED BY CHRONIC SKELETAL MUSCLE NECROSIS, LEADING TO MUSCLE REGENERATION FAILURE AND FIBROSIS. ALTHOUGH MACROPHAGES (MPS) ARE NORMALLY ESSENTIAL FOR MUSCLE REGENERATION, DYSREGULATED MP FUNCTION PROMOTES PATHOLOGICAL MUSCLE REMODELLING. INFILTRATING MPS CAN BE PREDOMINANTLY PRO-INFLAMMATORY (M1 BIASED), ANTI-INFLAMMATORY (M2 BIASED) OR OF A MIXED PHENOTYPE AND CAN ORIGINATE FROM THE ADULT BONE MARROW (MONOCYTE DEPENDENT) OR EMBRYONIC PRECURSORS (MONOCYTE INDEPENDENT). IN MDX MICE (GENETIC MODEL OF DMD) LACKING EITHER TOLL-LIKE RECEPTOR (TLR) 2 OR TLR4, IT IS FOUND THAT MP INFILTRATION OF DYSTROPHIC MUSCLE IS SIGNIFICANTLY REDUCED AND THAT THE MP PHENOTYPE IS SHIFTED TOWARD A MORE ANTI-INFLAMMATORY PROFILE. THIS IS ACCOMPANIED BY SIGNIFICANT IMPROVEMENTS IN MUSCLE HISTOLOGY AND FORCE PRODUCTION. LACK OF THE CHEMOKINE RECEPTOR CCR2, WHICH IMPEDES MONOCYTE RELEASE FROM THE BONE MARROW, LEADS TO SIMILAR BENEFICIAL EFFECTS IN MDX MICE. EVIDENCE WAS ALSO FOUND FOR TLR4-REGULATED INDUCTION OF TRAINED INNATE IMMUNITY IN MPS CULTURED FROM THE BONE MARROW OF MDX MICE BEFORE THEIR ENTRY INTO THE MUSCLE. THESE MPS EXHIBIT EPIGENETIC AND METABOLIC ALTERATIONS, ACCOMPANIED BY NON-SPECIFIC HYPER-RESPONSIVENESS TO MULTIPLE STIMULI, WHICH IS MANIFESTED BY POTENTIATED UPREGULATION OF BOTH PRO- AND ANTI-INFLAMMATORY GENES. IN SUMMARY, EXAGGERATED RECRUITMENT OF MONOCYTE-DERIVED MPS AND SIGNS OF TRAINED INNATE IMMUNITY AT THE LEVEL OF THE BONE MARROW ARE FEATURES OF THE IMMUNOPHENOTYPE ASSOCIATED WITH DYSTROPHIC MUSCLE DISEASE. THESE PHENOMENA ARE REGULATED BY TOLL-LIKE RECEPTORS THAT BIND ENDOGENOUS DAMAGE-ASSOCIATED MOLECULAR PATTERN (DAMP) MOLECULES, SUGGESTING THAT DAMP RELEASE FROM DYSTROPHIC MUSCLES MODULATES MP PLASTICITY AT THE BONE MARROW LEVEL THROUGH TOLL-LIKE RECEPTOR-DRIVEN MECHANISMS. 2022