1 875 132 CHRONIC AND TRANSIENT HYPERGLYCEMIA INDUCES CHANGES IN THE EXPRESSION PATTERNS OF IL6 AND ADIPOQ GENES AND THEIR ASSOCIATED EPIGENETIC MODIFICATIONS IN DIFFERENTIATING HUMAN VISCERAL ADIPOCYTES. ADIPOKINES SECRETED BY HYPERTROPHIC VISCERAL ADIPOSE TISSUE (VAT) INSTIGATE LOW-GRADE INFLAMMATION, FOLLOWED BY HYPERGLYCEMIA (HG)-RELATED METABOLIC DISORDERS. THE LATTER MAY DEVELOP WITH THE PARTICIPATION OF EPIGENETIC MODIFICATIONS. OUR AIM WAS TO ASSESS HOW HG INFLUENCES SELECTED EPIGENETIC MODIFICATIONS AND THE EXPRESSION OF INTERLEUKIN 6 (IL-6) AND ADIPONECTIN (APN; GENE SYMBOL ADIPOQ) DURING THE ADIPOGENESIS OF HUMAN VISCERAL PREADIPOCYTES (HPA-V). ADIPOCYTES (ADS) WERE CHRONICALLY OR TRANSIENTLY HG-TREATED DURING THREE STAGES OF ADIPOGENESIS (PROLIFERATION, DIFFERENTIATION, MATURATION). WE MEASURED ADIPOKINE MRNA, PROTEIN, PROVEN OR PREDICTED MICRORNA EXPRESSION (RT-QPCR AND ELISA), AND ENRICHMENT OF H3K9/14AC, H3K4ME3, AND H3K9ME3 AT GENE PROMOTER REGIONS (CHROMATIN IMMUNOPRECIPITATION). IN CHRONIC HG, WE DETECTED DIFFERENT EXPRESSION PATTERNS OF THE STUDIED ADIPOKINES AT THE MRNA AND PROTEIN LEVELS. CHRONIC AND TRANSIENT HG-INDUCED CHANGES IN MIRNA (MIR-26A-5P, MIR-26B-5P, LET-7D-5P, LET-7E-5P, MIR-365A-3P, MIR-146A-5P) WERE MOSTLY CONVERGENT TO ALTERED IL-6 TRANSCRIPTION. ALTERATIONS IN HISTONE MARKS AT THE IL6 PROMOTER WERE ALSO IN AGREEMENT WITH IL-6 MRNA. THE OPEN CHROMATIN MARKS AT THE ADIPOQ PROMOTER MOSTLY REFLECTED THE APN TRANSCRIPTION DURING NG ADIPOGENESIS, WHILE, IN THE DIFFERENTIATION STAGE, HG-INDUCED CHANGES IN ALL STUDIED MARKS WERE IN LINE WITH APN MRNA LEVELS. IN SUMMARY, HG DYSREGULATED ADIPOKINE EXPRESSION, PROMOTING INFLAMMATION. EPIGENETIC CHANGES COEXISTED WITH ALTERED EXPRESSION OF ADIPOKINES, ESPECIALLY FOR IL-6; THEREFORE, EPIGENETIC MARKS INDUCED BY TRANSIENT HG MAY ACT AS EPI-MEMORY IN ADS. SUCH CHANGES IN THE EPIGENOME AND EXPRESSION OF ADIPOKINES COULD BE INSTRUMENTAL IN THE DEVELOPMENT OF INFLAMMATION AND METABOLIC DEREGULATION OF VAT. 2021 2 2117 33 EPIGENETIC HISTONE METHYLATION MODULATES FIBROTIC GENE EXPRESSION. TGF-BETA1-INDUCED EXPRESSION OF EXTRACELLULAR MATRIX (ECM) GENES PLAYS A MAJOR ROLE IN THE DEVELOPMENT OF CHRONIC RENAL DISEASES SUCH AS DIABETIC NEPHROPATHY. ALTHOUGH MANY KEY TRANSCRIPTION FACTORS ARE KNOWN, MECHANISMS INVOLVING THE NUCLEAR CHROMATIN THAT MODULATE ECM GENE EXPRESSION REMAIN UNCLEAR. HERE, WE EXAMINED THE ROLE OF EPIGENETIC CHROMATIN MARKS SUCH AS HISTONE H3 LYSINE METHYLATION (H3KME) IN TGF-BETA1-INDUCED GENE EXPRESSION IN RAT MESANGIAL CELLS UNDER NORMAL AND HIGH-GLUCOSE (HG) CONDITIONS. TGF-BETA1 INCREASED THE EXPRESSION OF THE ECM-ASSOCIATED GENES CONNECTIVE TISSUE GROWTH FACTOR, COLLAGEN-ALPHA1[IOTA], AND PLASMINOGEN ACTIVATOR INHIBITOR-1. INCREASED LEVELS OF CHROMATIN MARKS ASSOCIATED WITH ACTIVE GENES (H3K4ME1, H3K4ME2, AND H3K4ME3), AND DECREASED LEVELS OF REPRESSIVE MARKS (H3K9ME2 AND H3K9ME3) AT THESE GENE PROMOTERS ACCOMPANIED THESE CHANGES IN EXPRESSION. TGF-BETA1 ALSO INCREASED EXPRESSION OF THE H3K4 METHYLTRANSFERASE SET7/9 AND RECRUITMENT TO THESE PROMOTERS. SET7/9 GENE SILENCING WITH SIRNAS SIGNIFICANTLY ATTENUATED TGF-BETA1-INDUCED ECM GENE EXPRESSION. FURTHERMORE, A TGF-BETA1 ANTIBODY NOT ONLY BLOCKED HG-INDUCED ECM GENE EXPRESSION BUT ALSO REVERSED HG-INDUCED CHANGES IN PROMOTER H3KME LEVELS AND SET7/9 OCCUPANCY. TAKEN TOGETHER, THESE RESULTS SHOW THE FUNCTIONAL ROLE OF EPIGENETIC CHROMATIN HISTONE H3KME IN TGF-BETA1-MEDIATED ECM GENE EXPRESSION IN MESANGIAL CELLS UNDER NORMAL AND HG CONDITIONS. PHARMACOLOGIC AND OTHER THERAPIES THAT REVERSE THESE MODIFICATIONS COULD HAVE POTENTIAL RENOPROTECTIVE EFFECTS FOR DIABETIC NEPHROPATHY. 2010 3 1897 31 ENDOTHELIN-1 REGULATION IS ENTANGLED IN A COMPLEX WEB OF EPIGENETIC MECHANISMS IN DIABETES. ENDOTHELIAL CELLS (ECS) ARE PRIMARY TARGETS OF GLUCOSE-INDUCED TISSUE DAMAGE. AS A RESULT OF HYPERGLYCEMIA, ENDOTHELIN-1 (ET-1) IS UPREGULATED IN ORGANS AFFECTED BY CHRONIC DIABETIC COMPLICATIONS. THE OBJECTIVE OF THE PRESENT STUDY WAS TO IDENTIFY NOVEL TRANSCRIPTIONAL MECHANISMS THAT INFLUENCE ET-1 REGULATION IN DIABETES. WE CARRIED OUT THE INVESTIGATION IN MICROVASCULAR ECS USING MULTIPLE APPROACHES. ECS WERE INCUBATED WITH 5 MM GLUCOSE (NG) OR 25 MM GLUCOSE (HG) AND ANALYSES FOR DNA METHYLATION, HISTONE METHYLATION, OR LONG NON-CODING RNA- MEDIATED REGULATION OF ET-1 MRNA WERE THEN PERFORMED. DNA METHYLATION ARRAY ANALYSES DEMONSTRATED THE PRESENCE OF HYPOMETHYLATION IN THE PROXIMAL PROMOTER AND 5' UTR/FIRST EXON REGIONS OF EDN1 FOLLOWING HG CULTURE. FURTHER, GLOBALLY BLOCKING DNA METHYLATION OR HISTONE METHYLATION SIGNIFICANTLY INCREASED ET-1 MRNA EXPRESSIONS IN BOTH NG AND HG-TREATED HRECS. WHILE, KNOCKING DOWN THE PATHOGENETIC LNCRNAS ANRIL, MALAT1, AND ZFAS1 SUBSEQUENTLY PREVENTED THE GLUCOSE-INDUCED UPREGULATION OF ET-1 TRANSCRIPTS. BASED ON OUR PAST AND PRESENT FINDINGS, WE PRESENT A NOVEL PARADIGM THAT REVEALS A COMPLEX WEB OF EPIGENETIC MECHANISMS REGULATING GLUCOSE-INDUCED TRANSCRIPTION OF ET-1. IMPROVING OUR UNDERSTANDING OF SUCH PROCESSES MAY LEAD TO BETTER TARGETED THERAPIES. 2018 4 916 25 CHRONIC HIGH GLUCOSE AND INSULIN STIMULATE BONE-MARROW STROMAL CELLS ADIPOGENIC DIFFERENTIATION IN YOUNG SPONTANEOUSLY HYPERTENSIVE RATS. WE EVALUATED WHETHER GENETIC PREDISPOSITION IS SUFFICIENT TO INDUCE CHANGES DUE TO CHRONIC HIGH GLUCOSE (HG; 25 MMOL/L) IN THE PRESENCE OR ABSENCE OF INSULIN (HGI; 10 MUG/ML) ON OSTEOGENIC DIFFERENTIATION AND MARKERS IN BONE-MARROW MESENCHYMAL STEM CELLS (BMSCS) FROM YOUNG WISTAR (WBMSCS) AND SPONTANEOUS HYPERTENSIVE RATS (SBMSCS) WITHOUT HYPERTENSION. HG SUPPRESSED OSTEOGENIC DIFFERENTIATION IN BOTH THE STRAINS, OBSERVED BY MINERALIZATION INHIBITION AND DECREASED LEVELS OF THE OSTEOGENIC MARKERS RUNX2, OSTERIX, OSTEOPONTIN, AND BONE SIALOPROTEIN, COMPARED TO OSTEOGENIC MEDIUM (OM) CELLS. IN WBMSCS, THE EFFECTS OF HG WERE ASSOCIATED WITH THE DOWN REGULATION OF ERK1/2 AND UP REGULATION OF P38 ACTIVITIES; HOWEVER, HGI DID NOT REVERT THE EFFECTS OF HG ON MAPK ACTIVITIES. MOREOVER, HG DID NOT AFFECT MAPK SIGNALING IN SBMSCS COMPARED TO THAT IN OM. HGI INCREASED MINERALIZATION IN WBMSCS COMPARED TO THAT IN OM, BUT NOT IN SBMSCS. HIGH EXPRESSION OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-GAMMA AND GLUCOSE TRANSPORTER TYPE 4 IN OM COULD BE RELATED WITH THE PREDISPOSITION TO ADIPOGENIC DIFFERENTIATION NOTED IN SBMSCS AND WAS CONFIRMED BY EMERGENCE OF ADIPOCYTE-LIKE CELLS BY HGI TREATMENT. DOWNREGULATION OF P38 AND UPREGULATION OF JNK ACTIVITIES WERE OBSERVED IN BOTH BMSCS TREATED WITH HGI COMPARED TO THOSE TREATED BY HG. MA (OSMOTIC CONTROL) ALSO SUPPRESSED OSTEOGENIC DIFFERENTIATION IN BOTH THE STRAINS. IN CONCLUSION, WE DEMONSTRATED THAT SBMSCS FROM YOUNG SPONTANEOUS HYPERTENSIVE RATS, WITHOUT HYPERTENSION BUT WITH GENETIC AND EPIGENETIC PREDISPOSITION, EXHIBITED DECREASED OSTEOBLASTIC DIFFERENTIATION UNDER HG AND HGI DID NOT REVERT THE EFFECTS OF HG IN SBMSCS BUT INCREASED ADIPOGENIC DIFFERENTIATION. 2018 5 6541 23 TRANSCRIPTOME ANALYSIS OF HUMAN PRIMARY ENDOTHELIAL CELLS (HUVEC) FROM UMBILICAL CORDS OF GESTATIONAL DIABETIC MOTHERS REVEALS CANDIDATE SITES FOR AN EPIGENETIC MODULATION OF SPECIFIC GENE EXPRESSION. WITHIN THE COMPLEX PATHOLOGICAL PICTURE ASSOCIATED TO DIABETES, HIGH GLUCOSE (HG) HAS "PER SE" EFFECTS ON CELLS AND TISSUES THAT INVOLVE EPIGENETIC REPROGRAMMING OF GENE EXPRESSION. IN FETAL TISSUES, EPIGENETIC CHANGES OCCUR GENOME-WIDE AND ARE BELIEVED TO INDUCE SPECIFIC LONG TERM EFFECTS. HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS (HUVEC) OBTAINED AT DELIVERY FROM GESTATIONAL DIABETIC WOMEN WERE USED TO STUDY THE TRANSCRIPTOMIC EFFECTS OF CHRONIC HYPERGLYCEMIA IN FETAL VASCULAR CELLS USING AFFYMETRIX MICROARRAYS. IN SPITE OF THE SMALL NUMBER OF SAMPLES ANALYZED (N=6), GENES RELATED TO INSULIN SENSING AND EXTRACELLULAR MATRIX REORGANIZATION WERE FOUND SIGNIFICANTLY AFFECTED BY HG. QUANTITATIVE PCR ANALYSIS OF GENE PROMOTERS IDENTIFIED A SIGNIFICANT DIFFERENTIAL DNA METHYLATION IN TGFB2. USE OF EA.HY926 ENDOTHELIAL CELLS CONFIRMS DATA ON HUVEC. OUR STUDY CORROBORATES RECENT EVIDENCES SUGGESTING THAT EPIGENETIC REPROGRAMMING OF GENE EXPRESSION OCCURS WITH PERSISTENT HG AND PROVIDES A BACKGROUND FOR FUTURE INVESTIGATIONS ADDRESSING GENOMIC CONSEQUENCES OF CHRONIC HG. 2014 6 6453 30 THIOREDOXIN INTERACTING PROTEIN (TXNIP) INDUCES INFLAMMATION THROUGH CHROMATIN MODIFICATION IN RETINAL CAPILLARY ENDOTHELIAL CELLS UNDER DIABETIC CONDITIONS. CHRONIC HYPERGLYCEMIA AND ACTIVATION OF RECEPTOR FOR ADVANCED GLYCATION END PRODUCTS (RAGE) ARE KNOWN RISK FACTORS FOR MICROVASCULAR DISEASE DEVELOPMENT IN DIABETIC RETINOPATHY. THIOREDOXIN-INTERACTING PROTEIN (TXNIP), AN ENDOGENOUS INHIBITOR OF ANTIOXIDANT THIOREDOXIN (TRX), PLAYS A CAUSATIVE ROLE IN DIABETES AND ITS VASCULAR COMPLICATIONS. HEREIN WE INVESTIGATE WHETHER HG AND RAGE INDUCE INFLAMMATION IN RAT RETINAL ENDOTHELIAL CELLS (EC) UNDER DIABETIC CONDITIONS IN CULTURE THROUGH TXNIP ACTIVATION AND WHETHER EPIGENETIC MECHANISMS PLAY A ROLE IN INFLAMMATORY GENE EXPRESSION. WE SHOW THAT RAGE ACTIVATION BY ITS LIGAND S100B OR HG TREATMENT OF RETINAL EC INDUCES THE EXPRESSION OF TXNIP AND INFLAMMATORY GENES SUCH AS COX2, VEGF-A, AND ICAM1. TXNIP SILENCING BY SIRNA IMPEDES RAGE AND HG EFFECTS WHILE STABLE OVER-EXPRESSION OF A CDNA FOR HUMAN TXNIP IN EC ELEVATES INFLAMMATION. P38 MAPK-NF-KAPPAB SIGNALING PATHWAY AND HISTONE H3 LYSINE (K) NINE MODIFICATIONS ARE INVOLVED IN TXNIP-INDUCED INFLAMMATION. CHROMATIN IMMUNOPRECIPITATION (CHIP) ASSAYS REVEAL THAT TXNIP OVER-EXPRESSION IN EC ABOLISHES H3K9 TRI-METHYLATION, A MARKER FOR GENE INACTIVATION, AND INCREASES H3K9 ACETYLATION, AN INDICATOR OF GENE INDUCTION, AT PROXIMAL COX2 PROMOTER BEARING THE NF-KAPPAB-BINDING SITE. THESE FINDINGS HAVE IMPORTANT IMPLICATIONS TOWARD UNDERSTANDING THE MOLECULAR MECHANISMS OF OCULAR INFLAMMATION AND ENDOTHELIAL DYSFUNCTION IN DIABETIC RETINOPATHY. 2009 7 6313 28 THE RELATIONSHIP BETWEEN MERCURY EXPOSURE AND EPIGENETIC ALTERATIONS REGARDING HUMAN HEALTH, RISK ASSESSMENT AND DIAGNOSTIC STRATEGIES. BACKGROUND: EXPOSURE TO THE ENVIRONMENTAL TOXICANTS POSES A SERIOUS THREAT TO HUMAN HEALTH. THE EXTENT OF EXPOSURE AND THE DEVELOPMENT OF DISEASES ARE INTERRELATED WITH EACH OTHER. CHRONIC EXPOSURE TO MERCURY (HG) INCREASES THE RISK OF DEVELOPING SERIOUS HUMAN DISORDERS FROM EMBRYO TO ADULTHOOD. OBJECTIVES: THE PURPOSE OF THIS REVIEW IS TO HIGHLIGHT THE MOST COMMON HUMAN DISORDERS INDUCED BY HG EXPOSURE ON THE BASIS OF EPIGENETIC MECHANISMS. A GROWING BODY OF EVIDENCE SHOWS THAT HG EXPOSURE LEADS TO ALTERATIONS IN THE EPIGENETIC MARKERS. METHODS: WE PERFORMED AN ORGANIZED SEARCH OF THE AVAILABLE LITERATURE USING PUBMED, GOOGLE SCHOLAR, MEDLINE, REAXYS, EMBASE AND SCOPUS DATABASES. ALL THE RELEVANT CITATIONS, INCLUDING RESEARCH AND REVIEW ARTICLES IN ENGLISH WERE EVALUATED. THE SEARCH TERMS INCLUDED MERCURY, HG, EPIGENETICS, EPIGENETIC ALTERATIONS, DNA METHYLATION, HISTONE MODIFICATIONS, MICRORNAS (MIRNAS), AND RISK ASSESSMENT. RESULTS: DATA ON HUMAN TOXICITY DUE TO HG EXPOSURE SHOWS BROAD VARIATIONS IN TERMS OF CHEMICAL NATURE, DOSES, AND THE RATE OF EXPOSURE. HG CONSUMPTION EITHER VIA FOODS OR ENVIRONMENTAL SOURCES MAY CREATE DELETERIOUS HEALTH EFFECTS ON VARIOUS PHYSIOLOGICAL SYSTEMS AT LEAST PARTIALLY THROUGH AN EPIGENETIC MECHANISM. CONCLUSION: HG EXPOSURE COULD TRIGGER EPIGENETIC ALTERATIONS, HENCE LEADING TO VARIOUS HUMAN DISORDERS INCLUDING REDUCED NEWBORN CEREBELLUM SIZE, ADVERSE BEHAVIORAL OUTCOMES, ATHEROSCLEROSIS AND MYOCARDIAL INFARCTION. SIMILARLY, IN ADULTS, OCCUPATIONAL HG EXPOSURE HAS BEEN ASSOCIATED WITH AN INCREASED RISK OF AUTOIMMUNITY. IT HAS BEEN REVEALED THAT MIRNAS IN THE WOMAN'S CERVIX ARE A NOVEL RESPONDER TO MATERNAL HG EXPOSURE DURING PREGNANCY. HG-INDUCED EPIGENETIC ALTERATIONS ANALYSIS OF KIDNEY TISSUES SHOWED A SIGNIFICANT INTERRUPTION IN RENAL FUNCTION. DNA METHYLATION AND HISTONE POST-TRANSLATION MODIFICATIONS ARE PREDOMINANT TYPES OF HG EPIGENETIC ALTERATIONS. 2019 8 858 28 CHROMATIN DECONDENSATION AND T CELL HYPERRESPONSIVENESS IN DIABETES-ASSOCIATED HYPERGLYCEMIA. DIABETES IS LINKED TO INCREASED INFLAMMATION AND SUSCEPTIBILITY TO CERTAIN INFECTIOUS DISEASES INCLUDING TUBERCULOSIS (TB). WE PREVIOUSLY REPORTED THAT AEROSOL TB IN MICE WITH CHRONIC (>/= 12 WK) HYPERGLYCEMIA FEATURES INCREASED BACTERIAL LOAD, OVERPRODUCTION OF SEVERAL CYTOKINES, AND INCREASED IMMUNE PATHOLOGY COMPARED WITH NORMOGLYCEMIC CONTROLS. A SIMILAR PHENOTYPE EXISTS IN HUMAN PATIENTS WITH DIABETES WITH TB. THE MECHANISMS OF INCREASED T CELL ACTIVATION IN DIABETES ARE UNKNOWN. IN THE CURRENT STUDY, WE TESTED THE HYPOTHESIS THAT HYPERGLYCEMIA MODIFIES THE INTRINSIC RESPONSIVENESS OF NAIVE T CELLS TO TCR STIMULATION. PURIFIED T CELLS FROM CHRONICALLY HYPERGLYCEMIC (HG) MICE PRODUCED HIGHER LEVELS OF TH1, TH2, AND TH17 CYTOKINES AND PROLIFERATED MORE THAN T CELLS FROM NORMOGLYCEMIC CONTROLS AFTER ANTI-CD3E OR AG STIMULATION. IN THIS WAY, NAIVE T CELLS FROM HG MICE RESEMBLED AG-EXPERIENCED CELLS, ALTHOUGH CD44 EXPRESSION WAS NOT INCREASED. CHROMATIN DECONDENSATION, ANOTHER CHARACTERISTIC OF AG-EXPERIENCED T CELLS, WAS INCREASED IN NAIVE T CELLS FROM HG MICE. THAT PHENOTYPE DEPENDED ON EXPRESSION OF THE RECEPTOR FOR ADVANCED GLYCATION END PRODUCTS AND COULD BE REVERSED BY INHIBITING P38 MAPK. CHROMATIN DECONDENSATION AND HYPERRESPONSIVENESS TO TCR STIMULATION PERSISTED FOLLOWING TRANSFER OF T CELLS FROM HG MICE INTO NORMOGLYCEMIC MICE. WE PROPOSE THAT CHRONIC HYPERGLYCEMIA CAUSES RECEPTOR FOR ADVANCED GLYCATION END PRODUCTS-MEDIATED EPIGENETIC MODIFICATION OF NAIVE T CELLS LEADING TO P38 MAPK-DEPENDENT CHROMATIN DECONDENSATION. THIS PREACTIVATION STATE FACILITATES TRANSCRIPTION FACTOR ACCESS TO DNA, INCREASING CYTOKINE PRODUCTION AND PROLIFERATION FOLLOWING TCR STIMULATION. THIS MECHANISM MAY CONTRIBUTE TO PATHOLOGICAL INFLAMMATION ASSOCIATED WITH DIABETES AND MIGHT OFFER A NOVEL THERAPEUTIC TARGET. 2014 9 3153 36 GLUCOSE-INDUCED EXPRESSION OF THE HOMEOTIC TRANSCRIPTION FACTOR PREP1 IS ASSOCIATED WITH HISTONE POST-TRANSLATIONAL MODIFICATIONS IN SKELETAL MUSCLE. AIMS/HYPOTHESIS: CHRONIC HYPERGLYCAEMIA WORSENS INSULIN RESISTANCE IN INDIVIDUALS WITH TYPE 2 DIABETES. WHETHER THIS EFFECT IS CONTRIBUTED BY EPIGENETIC DYSREGULATION AND WHICH GENES ARE INVOLVED REMAIN UNCLEAR. PREP1 (ALSO KNOWN AS PKNOX1) IS A GENE EXERTING MAJOR EFFECTS ON THE SENSITIVITY OF THE GLUCOSE TRANSPORT MACHINERY TO INSULIN. HERE, WE SHOW THAT DYSREGULATION OF PREP1 EXPRESSION BY HIGH GLUCOSE LEVELS IS ASSOCIATED WITH HISTONE MODIFICATIONS AT ITS 5' REGULATORY REGION. METHODS: WE USED MOUSE AND CELL MODELS TO INVESTIGATE PREP1 TRANSCRIPTIONAL REGULATION BY GLUCOSE. RESULTS: DIFFERENTIATED L6 SKELETAL MUSCLE CELLS WERE GROWN IN THE PRESENCE OF EITHER 5.5 OR 25 MMOL/L GLUCOSE (NORMAL [NG] AND HIGH GLUCOSE [HG], RESPECTIVELY). THE HG EXPOSURE INCREASED NUCLEAR FACTOR KAPPA LIGHT CHAIN ENHANCER OF ACTIVATED B CELLS (NF-KAPPAB) P65 BINDING AND RECRUITMENT OF THE SU(VAR)3-9, ENHANCER-OF-ZESTE, TRITHORAX DOMAIN-CONTAINING LYSINE METHYLTRANSFERASE 7 (SET7) HISTONE METHYLTRANSFERASE AND P300 ACETYLTRANSFERASE TO THE 5' REGION OF PREP1, LEADING TO ENHANCED TRANSCRIPTION. IN ADDITION, CHROMATIN IMMUNOPRECIPITATION ASSAYS REVEALED CONCOMITANTLY INCREASED HISTONE H3 MONO- AND DIMETHYLATION AND ACETYLATION AT LYS4 AND LYS9/14, RESPECTIVELY. SKELETAL MUSCLE TISSUE FROM STREPTOZOTOCIN-TREATED DIABETIC MICE ALSO SHOWED PREP1 OVEREXPRESSION ACCOMPANIED BY SIMILARLY INCREASED RECRUITMENT OF NF-KAPPAB P65 AND HISTONE MODIFICATIONS AT THE 5' REGION OF PREP1. IN THESE SAME MICE, AS WELL AS IN PREP1-OVEREXPRESSING L6 CELLS, PREP1-INDUCED RECRUITMENT OF THE REPRESSOR COMPLEX MYOCYTE ENHANCER FACTOR 2 (MEF2)/HISTONE DEACETYLASE 5 (HDAC5) AT THE GLUT4 PROMOTER WAS ALSO INCREASED, LEADING TO REDUCED GLUT4 EXPRESSION. CONCLUSIONS/INTERPRETATION: THESE STUDIES INDICATE THAT HG EXPOSURE INDUCES NF-KAPPAB RECRUITMENT AND HISTONE MODIFICATION AT THE PREP1 5' REGION, THEREBY ENHANCING THE TRANSCRIPTION OF PREP1 AND REPRESSING THAT OF GLUT4. HISTONE CHANGES AT THE PREP1 GENE MAY CONTRIBUTE TO INSULIN RESISTANCE IN INDIVIDUALS WITH TYPE 2 DIABETES. 2016 10 5186 19 PREMATURITY IN MICE LEADS TO REDUCTION IN NEPHRON NUMBER, HYPERTENSION, AND PROTEINURIA. THE NEPHRON NUMBER AT BIRTH IS A QUANTITATIVE TRAIT THAT CORRELATES INVERSELY WITH THE RISK OF HYPERTENSION AND CHRONIC KIDNEY DISEASE LATER IN LIFE. DURING KIDNEY DEVELOPMENT, THE NEPHRON NUMBER IS CONTROLLED BY MULTIPLE FACTORS INCLUDING GENETIC, EPIGENETIC, AND ENVIRONMENTAL MODIFIERS. PREMATURE BIRTH, WHICH REPRESENTS MORE THAN 12% OF ANNUAL LIVE BIRTHS IN THE UNITED STATES, HAS BEEN LINKED TO LOW NEPHRON NUMBER AND THE DEVELOPMENT OF HYPERTENSION LATER IN LIFE. IN THIS REPORT, WE DESCRIBE THE DEVELOPMENT OF A MOUSE MODEL OF PREMATURITY-INDUCED REDUCTION OF NEPHRON NUMBER. PREMATURE MICE, DELIVERED 1 AND 2 DAYS EARLY, HAVE 17.4 +/- 2.3% (N = 6) AND 23.6 +/- 2% (N = 10) FEWER NEPHRONS, RESPECTIVELY, WHEN COMPARED WITH FULL-TERM ANIMALS (12,252 +/- 571 NEPHRONS/KIDNEY, N = 10). AFTER 5 WEEKS OF AGE, THE MICE DELIVERED 2 DAYS PREMATURE SHOW LOWER REAL-TIME GLOMERULAR FILTRATION RATE (GFR, 283 +/- 13 VS 389 +/- 26 MUL/MIN). THE PREMATURE MICE ALSO DEVELOP HYPERTENSION (MEAN ARTERIAL PRESSURE [MAP], 134 +/- 18 VS 120 +/- 14 MM HG) AND ALBUMINURIA (286 +/- 83 VS 176 +/- 59 MUG ALBUMIN/MG CREATININE). THIS MOUSE MODEL PROVIDES A PROOF OF CONCEPT THAT PREMATURITY LEADS TO REDUCED NEPHRON NUMBER AND HYPERTENSION, AND THIS MODEL WILL BE USEFUL IN STUDYING THE PATHOPHYSIOLOGY OF PREMATURITY-INDUCED NEPHRON NUMBER REDUCTIONS AND HYPERTENSION. 2012 11 3149 35 GLUCOCORTICOID-INDUCED FINGERPRINTS ON VISCERAL ADIPOSE TISSUE TRANSCRIPTOME AND EPIGENOME. CONTEXT: CHRONIC GLUCOCORTICOID (GC) OVEREXPOSURE, RESULTING FROM ENDOGENOUS CUSHING'S SYNDROME (CS) OR EXOGENOUS GC THERAPY, CAUSES SEVERAL ADVERSE OUTCOMES, INCLUDING PERSISTENT CENTRAL FAT ACCUMULATION ASSOCIATED WITH A LOW-GRADE INFLAMMATION. HOWEVER, NO PREVIOUS MULTIOMICS STUDIES IN VISCERAL ADIPOSE TISSUE (VAT) FROM PATIENTS EXPOSED TO HIGH LEVELS OF UNSUPPRESSED GC DURING ACTIVE CS OR AFTER REMISSION ARE AVAILABLE YET. OBJECTIVE: TO DETERMINE THE PERSISTENT VAT TRANSCRIPTOMIC ALTERATIONS AND EPIGENETIC FINGERPRINTS INDUCED BY CHRONIC HYPERCORTISOLISM. METHODS: WE EMPLOYED A TRANSLATIONAL APPROACH COMBINING HIGH-THROUGHPUT DATA ON ENDOGENOUS CS PATIENTS AND A REVERSIBLE CS MOUSE MODEL. WE PERFORMED RNA SEQUENCING AND CHROMATIN IMMUNOPRECIPITATION SEQUENCING ON HISTONE MODIFICATIONS (H3K4ME3, H3K27AC, AND H3K27ME3) TO IDENTIFY PERSISTENT TRANSCRIPTIONAL AND EPIGENETIC SIGNATURES IN VAT PRODUCED DURING ACTIVE CS AND MAINTAINED AFTER REMISSION. RESULTS: VAT DYSFUNCTION WAS ASSOCIATED WITH LOW-GRADE PROINFLAMMATORY STATUS, MACROPHAGE INFILTRATION, AND EXTRACELLULAR MATRIX REMODELING. MOST NOTABLY, CHRONIC HYPERCORTISOLISM CAUSED A PERSISTENT CIRCADIAN RHYTHM DISRUPTION IN VAT THROUGH CORE CLOCK GENES MODULATION. IMPORTANTLY, CHANGES IN THE LEVELS OF 2 HISTONE MODIFICATIONS ASSOCIATED TO GENE TRANSCRIPTIONAL ACTIVATION (H3K4ME3 AND H3K27AC) CORRELATED WITH THE OBSERVED DIFFERENCES IN GENE EXPRESSION DURING ACTIVE CS AND AFTER CS REMISSION. CONCLUSION: WE IDENTIFIED FOR THE FIRST TIME THE PERSISTENT TRANSCRIPTIONAL AND EPIGENETIC SIGNATURES INDUCED BY HYPERCORTISOLISM IN VAT, PROVIDING A NOVEL INTEGRATED VIEW OF MOLECULAR COMPONENTS DRIVING THE LONG-TERM VAT IMPAIRMENT ASSOCIATED WITH CS. 2022 12 1725 36 DYSREGULATION OF INFLAMMATION, OXIDATIVE STRESS, AND GLUCOSE METABOLISM-RELATED GENES AND MIRNAS IN VISCERAL ADIPOSE TISSUE OF WOMEN WITH TYPE 2 DIABETES MELLITUS. BACKGROUND HUMAN VISCERAL ADIPOSE TISSUE (VAT), NOW IDENTIFIED AS AN ENDOCRINE ORGAN, PLAYS A SIGNIFICANT ROLE IN IMPAIRED FASTING GLUCOSE AND DIABETES THROUGH THE DEREGULATED METABOLISM AND ADIPOGENESIS OF VISCERAL ADIPOCYTES IN OBESITY. OUR STUDY FOCUSES ON EXPLORING THE LINK BETWEEN INFLAMMATION, OXIDATIVE STRESS, AND GLUCOSE METABOLISM-ASSOCIATED GENES WITH CORRESPONDING MIRNAS IN HUMAN VISCERAL ADIPOCYTES AND VAT FROM INDIVIDUALS WITH GLUCOSE METABOLISM DISORDERS. MATERIAL AND METHODS WE EXAMINED THE EXPRESSION OF ATM, NFKB1, SOD2, INSR, AND TIGAR, ALONG WITH THEIR RELATED MIRNAS USING PCR, IN TWO CONTEXTS:1 - DURING THE THREE-STAGE VISCERAL ADIPOGENESIS UNDER NORMAL GLUCOSE LEVELS (5.5 MILLIMOLES), INTERMITTENT, AND CHRONIC HYPERGLYCEMIA (30 MILLIMOLES).2 - IN VISCERAL ADIPOSE TISSUE FROM SUBJECTS (34 F, 18 M) WITH NORMAL GLUCOSE METABOLISM, IMPAIRED FASTING GLUCOSE, AND TYPE 2 DIABETES MELLITUS. RESULTS BOTH CHRONIC AND INTERMITTENT HYPERGLYCEMIA SIMILARLY INFLUENCED ATM, NFKB1, TIGAR, SOD2, INSR GENE EXPRESSION IN VISCERAL ADIPOCYTES, WITH CORRESPONDING CHANGES IN A FEW TESTED MIRNAS (EG, LET-7G-5P, MIR-145-5P, MIR-21-5P). ANTHROPOMETRIC AND BIOCHEMICAL PARAMETERS LED US TO FOCUS ON FEMALE SUBJECTS. OUR RESULTS SHOWED TRANSACTIVATION OF NFKB1, TIGAR, MIR-10B-5P, MIR-132-3P, MIR-20A-5P, MIR-21-5P, AND MIR-26A-5P EXCLUSIVELY IN TYPE 2 DIABETES MELLITUS. UPREGULATED MOLECULES (EXCLUDING MIR-10B-5P AND MIR-20A-5P) POSITIVELY CORRELATED WITH GLUCOSE METABOLISM MARKERS. CONCLUSIONS THE GENES STUDIED MAY UNDERGO MIRNA INTERFERENCES AND HYPERGLYCEMIC MEMORY IN VISCERAL ADIPOCYTES UNDER HYPERGLYCEMIC CONDITIONS. VAT FROM WOMEN WITH TYPE 2 DIABETES MELLITUS, BUT NOT WITH IMPAIRED FASTING GLUCOSE, SHOWED TRANSACTIVATED MIRNAS AND A MOLECULAR DYSREGULATION OF TIGAR AND NFKB1, POSSIBLY ENHANCING INFLAMMATION, OXIDATIVE STRESS, AND DISRUPTED GLUCOSE METABOLISM. THESE FINDINGS HIGHLIGHT THE EPIGENETIC AND MOLECULAR DISTURBANCES IN VAT RELATED TO GLUCOSE METABOLISM ABNORMALITIES. HOWEVER, ADDITIONAL RESEARCH IS NECESSARY TO FURTHER UNDERSTAND THEIR BIOLOGICAL SIGNIFICANCE. 2023 13 5675 24 SHIFTS IN PODOCYTE HISTONE H3K27ME3 REGULATE MOUSE AND HUMAN GLOMERULAR DISEASE. HISTONE PROTEIN MODIFICATIONS CONTROL FATE DETERMINATION DURING NORMAL DEVELOPMENT AND DEDIFFERENTIATION DURING DISEASE. HERE, WE SET OUT TO DETERMINE THE EXTENT TO WHICH DYNAMIC CHANGES TO HISTONES AFFECT THE DIFFERENTIATED PHENOTYPE OF ORDINARILY QUIESCENT ADULT GLOMERULAR PODOCYTES. TO DO THIS, WE EXAMINED THE CONSEQUENCES OF SHIFTING THE BALANCE OF THE REPRESSIVE HISTONE H3 LYSINE 27 TRIMETHYLATION (H3K27ME3) MARK IN PODOCYTES. ADRIAMYCIN NEPHROTOXICITY AND SUBTOTAL NEPHRECTOMY (SNX) STUDIES INDICATED THAT DELETION OF THE HISTONE METHYLATING ENZYME EZH2 FROM PODOCYTES DECREASED H3K27ME3 LEVELS AND SENSITIZED MICE TO GLOMERULAR DISEASE. H3K27ME3 WAS ENRICHED AT THE PROMOTER REGION OF THE NOTCH LIGAND JAG1 IN PODOCYTES, AND DEREPRESSION OF JAG1 BY EZH2 INHIBITION OR KNOCKDOWN FACILITATED PODOCYTE DEDIFFERENTIATION. CONVERSELY, INHIBITION OF THE JUMONJI C DOMAIN-CONTAINING DEMETHYLASES JMJD3 AND UTX INCREASED THE H3K27ME3 CONTENT OF PODOCYTES AND ATTENUATED GLOMERULAR DISEASE IN ADRIAMYCIN NEPHROTOXICITY, SNX, AND DIABETES. PODOCYTES IN GLOMERULI FROM HUMANS WITH FOCAL SEGMENTAL GLOMERULOSCLEROSIS OR DIABETIC NEPHROPATHY EXHIBITED DIMINISHED H3K27ME3 AND HEIGHTENED UTX CONTENT. ANALOGOUS TO HUMAN DISEASE, INHIBITION OF JMJD3 AND UTX ABATED NEPHROPATHY PROGRESSION IN MICE WITH ESTABLISHED GLOMERULAR INJURY AND REDUCED H3K27ME3 LEVELS. TOGETHER, THESE FINDINGS INDICATE THAT OSTENSIBLY STABLE CHROMATIN MODIFICATIONS CAN BE DYNAMICALLY REGULATED IN QUIESCENT CELLS AND THAT EPIGENETIC REPROGRAMMING CAN IMPROVE OUTCOMES IN GLOMERULAR DISEASE BY REPRESSING THE REACTIVATION OF DEVELOPMENTAL PATHWAYS. 2018 14 868 22 CHRONIC ADVANCED-GLYCATION END PRODUCTS TREATMENT INDUCES TXNIP EXPRESSION AND EPIGENETIC CHANGES IN GLOMERULAR PODOCYTES IN VIVO AND IN VITRO. ADVANCED GLYCATION END PRODUCTS (AGES) PLAY AN IMPORTANT ROLE IN OXIDATIVE STRESS AND INFLAMMATION, PROCESSES IMPLICATED IN THE DEVELOPMENT AND PROGRESSION OF KIDNEY DYSFUNCTION. IN THE PRESENT STUDY, WE INVESTIGATED THE PARTICIPATION OF THE PRO-OXIDANT PROTEIN THIOREDOXIN-INTERACTING PROTEIN (TXNIP) AND OF EPIGENETIC MECHANISMS ON KIDNEY TISSUE (IN VIVO, IN NON-DIABETIC RATS) AND ON TERMINALLY DIFFERENTIATED GLOMERULAR PODOCYTES (IN VITRO) CHRONICALLY EXPOSED TO AGES. AGES INDUCED TOTAL KIDNEY AND GLOMERULAR TXNIP EXPRESSION AND DECREASED H3K27ME3 CONTENT. CONCOMITANT TREATMENT WITH THE ANTIOXIDANT N-ACETYL-CYSTEINE (NAC) REVERSED ONLY THE INCREASED TXNIP EXPRESSION. TXNIP EXPRESSION POSITIVELY CORRELATED WITH PROTEINURIA AND NEGATIVELY CORRELATED WITH H3K27ME3 CONTENT. IN VITRO STUDIES IN PODOCYTES SHOWED THAT 72 H EXPOSURE TO AGES DECREASED NEPHRIN EXPRESSION AND INCREASED TXNIP, NOX4, COL4A1, AND EPITHELIAL-TO-MESENCHYMAL TRANSITION (EMT) MARKERS (ACTA2, SNAIL1, AND TGFB1). PODOCYTES TREATMENT WITH NAC REVERSED NOX4, COL4A1, ACTA2, AND TGFB1 INCREASED EXPRESSION BUT DID NOT ABROGATE THE REDUCED EXPRESSION OF NEPHRIN. MIR-29A EXPRESSION WAS DOWNREGULATED BY AGES IN VIVO, BUT NOT IN VITRO. IN CONCLUSION, TREATMENT OF NON-DIABETIC RATS WITH AGES INDUCED TXNIP EXPRESSION AND DECREASED THE CONTENTS OF THE REPRESSIVE EPIGENETIC MARK H3K27ME3 AND OF MIR-29A, POTENTIALLY DRIVING INJURY TO GLOMERULAR FILTRATION BARRIER AND PODOCYTES DYSFUNCTION. 2021 15 423 29 ANRIL REGULATES MULTIPLE MOLECULES OF PATHOGENETIC SIGNIFICANCE IN DIABETIC NEPHROPATHY. BACKGROUND: HYPERGLYCEMIA-INDUCED TRANSCRIPTIONAL ALTERATIONS LEAD TO ABERRANT SYNTHESIS OF A LARGE NUMBER OF PATHOGENETIC MOLECULES LEADING TO FUNCTIONAL AND STRUCTURAL DAMAGE TO MULTIPLE END ORGANS INCLUDING THE KIDNEYS. DIABETIC NEPHROPATHY (DN) REMAINS A MAJOR CAUSE OF END STAGE RENAL DISEASE. MULTIPLE EPIGENETIC MECHANISMS, INCLUDING ALTERATION OF LONG NON-CODING RNAS (LNCRNAS) MAY PLAY A SIGNIFICANT ROLE MEDIATING THE CELLULAR TRANSCRIPTIONAL ACTIVITIES. WE HAVE PREVIOUSLY SHOWN THAT LNCRNA ANRIL MAY MEDIATE DIABETES ASSOCIATED MOLECULAR, FUNCTIONAL AND STRUCTURAL ABNORMALITIES IN DN. HERE WE EXPLORED DOWNSTREAM MECHANISMS OF ANRIL ALTERATION IN DN. METHODS: WE USED RENAL CORTICAL TISSUES FROM ANRIL KNOCKOUT (KO) MICE AND WILD TYPE (WT) MICE, WITH OR WITHOUT STREPTOZOTOCIN (STZ) INDUCED DIABETES FOR RNA SEQUENCING. THE DIFFERENTIALLY EXPRESSED GENES WERE IDENTIFIED USING EDGER AND DESEQ2 COMPUTATIONAL METHODS. KEGG AND REACTOME PATHWAY ANALYSES AND NETWORK ANALYSES USING STRING AND IPA WERE SUBSEQUENTLY PERFORMED. RESULTS: DIABETIC ANIMALS SHOWED HYPERGLYCEMIA, REDUCED BODY WEIGHT GAIN, POLYURIA AND INCREASED URINARY ALBUMIN. BOTH ALBUMINURIA AND POLYURIA WERE CORRECTED IN THE KO DIABETIC MICE. RNA ANALYSES SHOWED DIABETES INDUCED ALTERATIONS OF A LARGE NUMBER OF TRANSCRIPTS IN THE WILD TYPE (WT) ANIMALS. ANRIL KNOCKOUT (KO) PREVENTED A LARGE NUMBER OF SUCH ALTERATIONS. THE ALTERED TRANSCRIPTS INCLUDE METABOLIC PATHWAYS, APOPTOSIS, EXTRACELLULAR MATRIX PROTEIN SYNTHESIS AND DEGRADATION, NFKB RELATED PATHWAYS, AGE-RAGE INTERACTION PATHWAYS ETC. ANRIL KO PREVENTED MAJORITY OF THESE PATHWAYS. CONCLUSION: THESE FINDINGS SUGGEST THAT AS ANRIL REGULATES A LARGE NUMBER OF MOLECULES OF PATHOGENETIC SIGNIFICANCE, IT MAY POTENTIALLY BE A DRUG TARGET FOR DN AND OTHER CHRONIC DIABETIC COMPLICATIONS. 2022 16 364 26 AMELIORATION OF UREMIC TOXIN INDOXYL SULFATE-INDUCED OSTEOBLASTIC CALCIFICATION BY SET DOMAIN CONTAINING LYSINE METHYLTRANSFERASE 7/9 PROTEIN. BACKGROUND: VASCULAR CALCIFICATION (VC) IS A VERY COMMON PHENOMENON IN PATIENTS WITH CHRONIC KIDNEY DISEASE (CKD). IT HAS BEEN REPORTED THAT SOME HISTONE METHYLATION PLAY A ROLE IN VC AS AN EPIGENETIC REGULATOR. INDOXYL SULFATE (IS) IS A PROTEIN-BOUND UREMIC TOXIN THAT HAS BEEN PROVEN AS ONE OF THE MAJOR RISK FACTORS OF CARDIOVASCULAR DISEASE IN CKD. SET DOMAIN CONTAINING LYSINE METHYLTRANSFERASE 7/9 (SET7/9) IS ONE OF THE IMPORTANT HISTONE METHYLTRANSFERASES. OBJECTIVES: THIS STUDY AIMED TO DETERMINE THE EFFECT OF IS ON THE EXPRESSION OF SET7/9 AND THE ROLE OF SET7/9 IN IS-INDUCED OSTEOBLASTIC DIFFERENTIATION AND CALCIFICATION OF VASCULAR SMOOTH MUSCLE CELLS (VSMCS). METHODS: VSMCS WERE INCUBATED WITH VARIOUS CONCENTRATIONS OF IS FOR DIFFERENT DURATIONS TO ASSESS OSTEOBLASTIC DIFFERENTIATION AND EXPRESSION OF SET7/9. WESTERN BLOT ANALYSIS AND QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION WERE PERFORMED TO ASSESS THE PROTEIN AND MRNA LEVELS OF SET7/9 RESPECTIVELY. THE CALCIUM CONTENT WAS MEASURED TO EVALUATE CALCIFICATION. RESULTS: OSTEOBLASTIC DIFFERENTIATION AND CALCIFICATION OF VSMCS AND DOWNREGULATION OF THE EXPRESSION OF SET7/9 WERE OBSERVED AFTER IS TREATMENT. THE AUTOPHAGY WAS ACTIVATED AFTER TREATMENT WITH IS, WHEREAS THE INHIBITION OF THE AUTOPHAGY PARTIALLY ATTENUATED THE EFFECT OF IS ON BOTH THE STIMULATION OF THE EXPRESSION OF RUNT-RELATED TRANSCRIPTION FACTOR 2 AND CALCIUM DEPOSITION. CONCLUSIONS: OUR DATA DEMONSTRATED THAT SET7/9 DOWNREGULATION AND AUTOPHAGY ACTIVATION MAY BE THE KEY MECHANISM OF IS-INDUCED VC IN CKD. 2019 17 273 29 AGE-INDUCED SUPPRESSION OF EZH2 MEDIATES INJURY OF PODOCYTES BY REDUCING H3K27ME3. BACKGROUND: CHRONIC HYPERGLYCEMIA, A PIVOTAL FEATURE OF DIABETES MELLITUS (DM), INITIATES THE FORMATION OF ADVANCED GLYCATION END PRODUCTS (AGES) AND THE DYSREGULATION OF EPIGENETIC MECHANISMS, WHICH MAY CAUSE INJURY TO RENAL PODOCYTES, A CENTRAL FEATURE OF DIABETIC KIDNEY DISEASE (DKD). PREVIOUS DATA OF OUR GROUP SHOWED THAT AGES SIGNIFICANTLY REDUCE THE EXPRESSION OF NIPP1 (NUCLEAR INHIBITOR OF PROTEIN PHOSPHATASE 1) IN PODOCYTES IN VITRO AS WELL AS IN HUMAN AND MURINE DKD. NIPP1 WAS SHOWN BY OTHERS TO INTERACT WITH ENHANCER OF ZESTE HOMOLOG 2 (EZH2), WHICH CATALYZES THE REPRESSIVE METHYLATION OF H3K27ME3 ON HISTONE 3. THEREFORE, WE HYPOTHESIZED THAT AGES CAN DIRECTLY INDUCE EPIGENETIC CHANGES IN PODOCYTES. METHODS: WE ANALYZED THE RELEVANCE OF AGES ON EZH2 EXPRESSION AND ACTIVITY IN A MURINE PODOCYTE CELL LINE. CELLS WERE TREATED WITH 5 MG/ML GLYCATED BSA FOR 24 H. TO DETERMINE THE MEANING OF EZH2 SUPPRESSION, EZH2 ACTIVITY WAS INHIBITED BY INCUBATING THE CELLS WITH THE PHARMACOLOGICAL METHYLTRANSFERASE INHIBITOR 3-DEAZANEPLANOCIN A; EZH2 EXPRESSION WAS REPRESSED WITH SIRNA. MRNA EXPRESSION WAS ANALYZED WITH REAL-TIME PCR, AND PROTEIN EXPRESSION WITH WESTERN BLOT. EZH2 EXPRESSION AND LEVEL OF H3K27 TRIMETHYLATION IN PODOCYTES OF DIABETIC DB/DB MICE, A MOUSE MODEL FOR TYPE 2 DM, WERE ANALYZED USING IMMUNOFLUORESCENCE. RESULTS: OUR DATA DEMONSTRATED THAT AGES DECREASE EZH2 EXPRESSION IN PODOCYTES AND CONSEQUENTLY REDUCE H3K27ME3. THIS SUPPRESSION OF EZH2 MIMICKED THE AGE EFFECTS AND CAUSED AN UPREGULATED EXPRESSION OF PATHOLOGICAL FACTORS THAT CONTRIBUTE TO PODOCYTE INJURY IN DKD. IN ADDITION, ANALYSES OF DB/DB MICE SHOWED SIGNIFICANTLY REDUCED H3K27ME3 AND EZH2 EXPRESSION IN PODOCYTES. MOREOVER, THE SUPPRESSION OF NIPP1 AND EZH2 SHOWED SIMILAR EFFECTS REGARDING PODOCYTE INJURY. CONCLUSIONS: OUR STUDIES PROVIDE A NOVEL PATHWAY HOW AGES CONTRIBUTE TO PODOCYTE INJURY AND THE FORMATION OF THE SO-CALLED METABOLIC MEMORY IN DKD. 2020 18 2149 22 EPIGENETIC MARKERS TO PREDICT CONVERSION FROM GESTATIONAL DIABETES TO TYPE 2 DIABETES. CONTEXT: LIFESTYLE FACTORS MEDIATE EPIGENETIC CHANGES THAT CAN CAUSE CHRONIC DISEASES. ALTHOUGH ANIMAL AND LABORATORY STUDIES LINK EPIGENETIC CHANGES TO DIABETES, EPIGENETIC INFORMATION IN WOMEN WITH GESTATIONAL DIABETES (GDM) AND TYPE 2 DIABETES IS LACKING. OBJECTIVE: THIS STUDY SOUGHT TO MEASURE EPIGENETIC MARKERS ACROSS PREGNANCY AND EARLY POSTPARTUM AND IDENTIFY MARKERS THAT COULD BE USED AS PREDICTORS FOR CONVERSION FROM GDM TO TYPE 2 DIABETES. DESIGN: GLOBAL HISTONE H3 DIMETHYLATION WAS MEASURED IN WHITE BLOOD CELLS AT THREE TIME POINTS: 30 WK GESTATION, 8-10 WK POSTPARTUM, AND 20 WK POSTPARTUM, FROM FOUR GROUPS OF WOMEN WITH AND WITHOUT DIABETES. SETTING AND PARTICIPANTS: A TOTAL OF 39 PARTICIPANTS (SIX TO NINE IN EACH GROUP) WERE RECRUITED INCLUDING: NONDIABETIC WOMEN; WOMEN WITH GDM WHO DEVELOPED POSTPARTUM TYPE 2 DIABETES; WOMEN WITH GDM WITHOUT POSTPARTUM TYPE 2 DIABETES; AND WOMEN WITH TYPE 2 DIABETES. MAIN OUTCOME MEASURE: PERCENTAGES OF DIMETHYLATION OF H3 HISTONES RELATIVE TO TOTAL H3 HISTONE METHYLATION WERE COMPARED BETWEEN DIABETIC/NONDIABETIC GROUPS USING APPROPRIATE COMPARATIVE STATISTICS. RESULTS: H3K27 DIMETHYLATION WAS 50-60% LOWER AT 8-10 AND 20 WK POSTPARTUM IN WOMEN WITH GDM WHO DEVELOPED TYPE 2 DIABETES, COMPARED WITH NONDIABETIC WOMEN. H3K4 DIMETHYLATION WAS 75% LOWER AT 8-10 WK POSTPARTUM IN WOMEN WITH GDM WHO SUBSEQUENTLY DEVELOPED TYPE 2 DIABETES COMPARED WITH WOMEN WHO HAD GDM WHO DID NOT. CONCLUSIONS: THE PERCENTAGE OF DIMETHYLATION OF HISTONES H3K27 AND H3K4 VARIED WITH DIABETIC STATE AND HAS THE POTENTIAL AS A PREDICTIVE TOOL TO IDENTIFY WOMEN WHO WILL CONVERT FROM GDM TO TYPE 2 DIABETES. 2016 19 4076 30 MATERNAL HIGH-FAT DIET MODIFIES EPIGENETIC MARKS H3K27ME3 AND H3K27AC IN BONE TO REGULATE OFFSPRING OSTEOBLASTOGENESIS IN MICE. STUDIES FROM BOTH HUMANS AND ANIMAL MODELS INDICATED THAT MATERNAL CHRONIC POOR-QUALITY DIET, ESPECIALLY A HIGH FAT DIET (HFD), IS SIGNIFICANTLY ASSOCIATED WITH REDUCED BONE DENSITY AND CHILDHOOD FRACTURES IN OFFSPRING. WHEN PREVIOUSLY STUDIED IN A RAT MODEL, OUR DATA SUGGESTED THAT MATERNAL HFD CHANGES EPIGENETIC MARKS SUCH AS DNA METHYLATION AND HISTONE MODIFICATIONS TO CONTROL OSTEOBLAST METABOLISM. IN MOUSE EMBRYONIC AND POSTNATAL OFFSPRING BONE SAMPLES, A CHIP-SEQUENCING (CHIP-SEQ)-BASED GENOME-WIDE METHOD WAS USED TO LOCATE THE REPRESSIVE HISTONE MARK H3K27ME3 (MEDIATED VIA THE POLYCOMB HISTONE METHYLTRANSFERASE, EZH2) AND EXPRESSIVE HISTONE MARK H3K27AC (P300/CBP MEDIATED) THROUGHOUT THE GENOME. USING ISOLATED MOUSE EMBRYONIC CELLS FROM FOETAL CALVARIA (OSTEOBLAST-LIKE CELLS), H3K27ME3 CHIP-SEQ SHOWED THAT 147 GENE BODIES AND 26 GENE PROMOTERS IN HFD EMBRYOTIC SAMPLES HAD A GREATER THAN TWOFOLD INCREASE IN H3K27ME PEAKS COMPARED TO CONTROLS. AMONG THE HFD SAMPLES, PTHLH AND COL2A1 THAT ARE IMPORTANT GENES PLAYING ROLES DURING CHONDRO- AND OSTEOGENESIS HAD SIGNIFICANTLY ENRICHED LEVELS OF H3K27ME3. THEIR DECREASED MRNA EXPRESSION WAS CONFIRMED BY REAL-TIME PCR AND STANDARD CHIP ANALYSIS, INDICATING A STRONG ASSOCIATION WITH EZH2 MEDIATED H3K27ME3 EPIGENETIC CHANGES. USING EMBRYONIC CALVARIA OSTEOBLASTIC CELLS AND OFFSPRING BONE SAMPLES, H3K27AC CHIP-SEQ ANALYSIS SHOWED THAT OSTEOBLAST INHIBITOR GENES TNFAIP3 AND TWIST1 HAD SIGNIFICANTLY ENRICHED PEAKS OF H3K27AC IN HFD SAMPLES COMPARED TO CONTROLS. THEIR INCREASED GENE EXPRESSION AND ASSOCIATION WITH H3K27AC WERE ALSO CONFIRMED BY REAL-TIME PCR AND STANDARD CHIP ANALYSIS. THESE FINDINGS INDICATE THAT CHRONIC MATERNAL HFD CHANGES HISTONE TRIMETHYLATION AND ACETYLATION EPIGENETIC MARKS TO REGULATE EXPRESSION OF GENES CONTROLLING OSTEOBLASTOGENESIS. 2022 20 4354 27 MIR-21-5P DIRECTLY CONTRIBUTES TO REGULATING ENOS EXPRESSION IN HUMAN ARTERY ENDOTHELIAL CELLS UNDER NORMOXIA AND HYPOXIA. CLINICAL CONDITIONS ASSOCIATED WITH HYPOXIA AND OXIDATIVE STRESS, SUCH AS FETAL GROWTH RESTRICTION (FGR), RESULTS IN ENDOTHELIAL DYSFUNCTION. PREVIOUS REPORTS SHOW THAT CHANGES IN ENOS EXPRESSION UNDER THESE CONDITIONS ARE TIGHTLY CONTROLLED BY DNA METHYLATION AND HISTONE POSTTRANSLATIONAL MODIFICATIONS. HOWEVER, THE CONTRIBUTION OF AN ORCHESTRATING EPIGENETIC MECHANISM, SUCH AS MIRNAS, ON THE NO-RELATED GENES EXPRESSION HAS NOT BEEN ADDRESSED. WE AIMED TO DETERMINE THE LEVELS OF MIRNAS HIGHLY EXPRESSED IN NORMAL ENDOTHELIAL CELLS (EC), MIR-21 AND MIR-126, IN FGR HUMAN UMBILICAL ARTERY EC (HUAEC), AND THEIR EFFECTS ON HYPOXIA-DEPENDENT REGULATION OF BOTH, NO-RELATED AND OXIDATIVE STRESS-RELATED GENES. RESULTS WERE VALIDATED BY TRANSCRIPTOME ANALYSIS OF HUAEC CULTURED UNDER CHRONIC LOW OXYGEN CONDITIONS. CULTURED FGR-HUAEC SHOWED DECREASED HSA-MIR-21, DDAH1, SOD1, AND NRF2, BUT INCREASED MIR-126, NOX4, AND ENOS LEVELS, COMPARED WITH CONTROLS. MIR-21-5P LEVELS IN FGR WERE ASSOCIATED WITH INCREASED HG-MIR-21 GENE PROMOTER METHYLATION, WITH NO CHANGES IN HG-MIR-126 GENE PROMOTER METHYLATION. HUAEC EXPOSED TO HYPOXIA SHOWED A TRANSIENT INCREASE IN ENOS AND DDAH11, PARALLELED BY DECREASE MIR-21-5P LEVELS, BUT NO CHANGES IN MIR-126-3P AND THE OTHER GENES UNDER STUDY. TRANSCRIPTOME PROFILING SHOWED AN INVERSE RELATIONSHIP AMONG MIR-21 AND SEVERAL TRANSCRIPTS TARGETED BY MIR-21 IN HUAEC EXPOSED TO HYPOXIA, MEANWHILE MIR-21-5P-MIMIC DECREASED ENOS AND DDAH1 TRANSCRIPTS STABILITY, BLOCKING THEIR INDUCTION BY HYPOXIA. CONSEQUENTLY, FGR PROGRAMS A HYPOXIA-RELATED MIRNA THAT CONTRIBUTES TO THE REGULATION OF THE NO PATHWAY, INVOLVING A DIRECT EFFECT OF MIR-21-5P ON ENOS TRANSCRIPT STABILITY, NOT PREVIOUSLY REPORTED. MOREOVER, HYPOXIA DOWNREGULATES MIR-21-5P, CONTRIBUTING TO INCREASING THE EXPRESSION OF NO-RELATED GENES IN ARTERIAL ENDOTHELIAL CELLS. 2020