1  4947 181 PATERNAL SEPSIS INDUCES ALTERATIONS OF THE SPERM METHYLOME AND DAMPENS OFFSPRING IMMUNE RESPONSES-AN ANIMAL STUDY. BACKGROUND: SEPSIS REPRESENTS THE UTMOST SEVERE CONSEQUENCE OF INFECTION, INVOLVING A DYSREGULATED AND SELF-DAMAGING IMMUNE RESPONSE OF THE HOST. WHILE DIFFERENT ENVIRONMENTAL EXPOSURES LIKE CHRONIC STRESS OR MALNUTRITION HAVE BEEN WELL DESCRIBED TO REPROGRAM THE GERMLINE AND SUBSEQUENTLY OFFSPRING ATTRIBUTES, THE INTERGENERATIONAL IMPACT OF SEPSIS AS A TREMENDOUS IMMUNOLOGICAL STRESSOR HAS NOT BEEN EXAMINED YET. METHODS: POLYMICROBIAL SEPSIS IN 12-WEEK-OLD MALE C57BL/6 MICE WAS INDUCED BY CECAL LIGATION AND PUNCTURE (CLP), FOLLOWED BY A MATING OF THE MALE SURVIVORS (OR APPROPRIATE SHAM CONTROL ANIMALS) 6 WEEKS LATER WITH HEALTHY FEMALES. ALVEOLAR MACROPHAGES OF OFFSPRING ANIMALS WERE ISOLATED AND STIMULATED WITH EITHER LPS OR ZYMOSAN, AND SUPERNATANT LEVELS OF TNF-ALPHA WERE QUANTIFIED BY ELISA. FURTHERMORE, SYSTEMIC CYTOKINE RESPONSE TO INTRAPERITONEALLY INJECTED LPS WAS ASSESSED AFTER 24 H. ALSO, MORPHOLOGY, MOTILITY, AND GLOBAL DNA METHYLATION OF THE SEPSIS SURVIVORS' SPERM WAS EXAMINED. RESULTS: COMPARATIVE REDUCED REDUCTION BISULFITE SEQUENCING (RRBS) OF SPERM REVEALED CHANGES OF DNA METHYLATION (N = 381), MOST PRONOUNCED IN THE INTERGENIC GENOME AS WELL AS WITHIN INTRONS OF DEVELOPMENTALLY RELEVANT GENES. OFFSPRING OF SEPSIS FATHERS EXHIBITED A SLIGHT DECREASE IN BODY WEIGHT, WITH A MORE PRONOUNCED WEIGHT DIFFERENCE IN MALE ANIMALS (CLP VS. SHAM). MALE DESCENDANTS OF SEPSIS FATHERS, BUT NOT FEMALE DESCENDANTS, EXHIBITED LOWER PLASMA CONCENTRATIONS OF IL-6, TNF-ALPHA, AND IL-10 24 H AFTER INJECTION OF LPS. IN LINE, ONLY ALVEOLAR MACROPHAGES OF MALE DESCENDANTS OF SEPSIS FATHERS PRODUCED LESS TNF-ALPHA UPON ZYMOSAN STIMULATION COMPARED TO SHAM DESCENDANTS, WHILE LPS RESPONSES KEPT UNCHANGED. CONCLUSION: WE CAN PROVE THAT MALE-BUT SURPRISINGLY NOT FEMALE-DESCENDANTS OF POST-SEPSIS FATHERS SHOW A DAMPENED SYSTEMIC AS WELL AS PULMONARY IMMUNE RESPONSE. BASED ON THIS OBSERVATION OF AN IMMUNE HYPO-RESPONSIVITY, WE PROPOSE THAT MALE DESCENDANTS OF SEPSIS FATHERS ARE AT RISK TO DEVELOP FUNGAL AND BACTERIAL INFECTIONS AND MIGHT BENEFIT FROM THERAPEUTIC IMMUNE MODULATION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                              
2   946  35 CHRONIC MATERNAL PROTEIN DEPRIVATION IN MICE IS ASSOCIATED WITH OVEREXPRESSION OF THE COHESIN-MEDIATOR COMPLEX IN LIVER OF THEIR OFFSPRING. EPIGENETIC MECHANISMS MAY PLAY AN IMPORTANT ROLE IN THE DEVELOPMENTAL PROGRAMMING OF ADULT-ONSET CHRONIC METABOLIC DISEASES RESULTING FROM SUBOPTIMAL FETAL NUTRITION, BUT THE EXACT MOLECULAR MECHANISMS ARE INCOMPLETELY UNDERSTOOD. GIVEN THE CENTRAL ROLE OF THE LIVER IN METABOLIC REGULATION, WE INVESTIGATED WHETHER CHRONIC MATERNAL DIETARY PROTEIN RESTRICTION HAS LONG-TERM EFFECTS ON LIVER GENE EXPRESSION IN THE OFFSPRING. WE FED ADULT C57BL/6J DAMS AD LIBITUM AN 8% MATERNAL LOW-PROTEIN (MLP) OR 20% PROTEIN CONTROL DIET (C) FROM 4 WK PRIOR TO MATING UNTIL THE END OF LACTATION. MALE PUPS WERE WEANED TO STANDARD NONPURIFIED DIET AND SINGLY HOUSED AT 21 D OF AGE (D 21). BODY WEIGHTS WERE FOLLOWED TO 1 Y OF AGE (1 Y). AT D 21 AND 1 Y, ORGANS WERE QUANTITATIVELY DISSECTED AND ANALYZED. MLP OFFSPRING HAD SIGNIFICANTLY LOWER BODY WEIGHTS AT ALL AGES AND SIGNIFICANTLY LOWER SERUM ACTIVITY OF ALANINE AMINOTRANSFERASE AND LACTATE DEHYDROGENASE AT 1 Y. GENE EXPRESSION PROFILING OF LIVER AT 1 Y SHOWED 521 OVEREXPRESSED AND 236 UNDEREXPRESSED GENES IN MLP COMPARED TO C OFFSPRING. THE MOST IMPORTANT NOVEL FINDING WAS THE OVEREXPRESSION OF GENES FOUND IN LIVER THAT PARTICIPATE IN ORGANIZATION AND MAINTENANCE OF HIGHER ORDER CHROMATIN ARCHITECTURE AND REGULATION OF TRANSCRIPTIONAL ACTIVATION. THESE INCLUDED MEMBERS OF THE COHESIN-MEDIATOR COMPLEX, WHICH REGULATE GENE EXPRESSION BY FORMING DNA LOOPS BETWEEN PROMOTERS AND ENHANCERS IN A CELL TYPE-SPECIFIC FASHION. THUS, OUR FINDINGS OF INCREASED EXPRESSION OF THESE FACTORS IN LIVER OF MLP OFFSPRING IMPLICATE A POSSIBLE NOVEL EPIGENETIC MECHANISM IN DEVELOPMENTAL PROGRAMMING.	2011	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
3  2312  39 EPIGENETIC REGULATION OF DENDRITIC CELL-DERIVED INTERLEUKIN-12 FACILITATES IMMUNOSUPPRESSION AFTER A SEVERE INNATE IMMUNE RESPONSE. PATIENTS WHO SURVIVE SEPSIS HAVE SIGNIFICANT DEFICIENCIES IN THEIR IMMUNE RESPONSES CAUSED BY POORLY UNDERSTOOD MECHANISMS. WE HAVE EXPLORED THIS PHENOMENON BY STUDYING DENDRITIC CELLS (DCS) RECOVERED FROM ANIMALS SURVIVING SEVERE PERITONITIS-INDUCED SEPSIS, USING THE WELL-ESTABLISHED CECAL LIGATION AND PUNCTURE (CLP) MODEL. IMMEDIATELY AFTER THE INITIATION OF SEPSIS THERE IS A DEPLETION IN DCS FROM THE LUNG AND SPLEEN, WHICH IS FOLLOWED BY REPOPULATION OF THESE CELLS BACK TO THE RESPECTIVE ORGANS. DCS RECOVERED FROM SURVIVING ANIMALS EXHIBITED A SIGNIFICANT AND CHRONIC SUPPRESSION OF INTERLEUKIN-12 (IL-12), A KEY HOST DEFENSE CYTOKINE. THE SUPPRESSION OF DC-DERIVED IL-12 PERSISTED FOR AT LEAST 6 WEEKS AFTER CLP AND WAS NOT DUE TO IMMUNOREGULATORY CYTOKINES, SUCH AS IL-10. USING CHROMATIN IMMUNOPRECIPITATION (CHIP) TECHNIQUES, WE HAVE SHOWN THAT THE DEFICIENCY IN DC-DERIVED IL-12 WAS DUE TO EPIGENETIC ALTERATIONS. SPECIFICALLY, IL-12 EXPRESSION WAS REGULATED BY STABLE RECIPROCAL CHANGES IN HISTONE H3 LYSINE-4 TRIMETHYLATION (H3K4ME3) AND HISTONE H3 LYSINE-27 DIMETHYLATION (H3K27ME2), AS WELL AS CHANGES IN COGNATE HISTONE METHYLTRANSFERASE (HMT) COMPLEXES ON THE IL12P35 AND IL12P40 PROMOTERS. THESE DATA IMPLICATE HISTONE MODIFICATION ENZYMES IN SUPPRESSING DC-DERIVED IL-12, WHICH MAY PROVIDE ONE OF THE MECHANISMS OF LONG-TERM IMMUNOSUPPRESSION SUBSEQUENT TO THE SEPTIC RESPONSE.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
4  4081  40 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
5  3785  38 INTERGENERATIONAL EFFECTS OF PRE-PREGNANCY CHRONIC LIPOPOLYSACCHARIDE FROM PORPHYROMONAS GINGIVALIS ON THE LEARNING, MEMORY AND SEIZURE SUSCEPTIBILITY OF OFFSPRING. OBJECTIVE: THE AIM OF THIS STUDY WAS TO INVESTIGATE THE EFFECTS OF PRE-PREGNANCY CHRONIC EXPOSURE TO PORPHYROMONAS GINGIVALIS LPS (PG LPS) ON THE LEARNING, MEMORY, AND SEIZURE SUSCEPTIBILITY OF THE OFFSPRING. DESIGN: TO ACHIEVE PERIODONTITIS, PG LPS (5 MUG/KG) WAS INJECTED INTO THE GINGIVAL OF FIVE FEMALE RATS EVERY 48 H FOR THREE WEEKS. FIVE CONTROL FEMALE RATS RECEIVED SALINE (0.9 %) AND FIVE FEMALE WERE KEPT INTACT. THE CONCENTRATIONS OF TNF-ALPHA AND IL-6 WERE MEASURED IN THE BLOOD SAMPLES. ONE WEEK AFTER THE FINAL INJECTION, FEMALES WERE MATED WITH INTACT MALES. FOLLOWING BIRTH AND WEANING, TWO MALE AND TWO FEMALE OFFSPRING WERE RANDOMLY SELECTED FROM EACH MOTHER, AND NEW GROUPS OF MALE AND FEMALE OFFSPRING WERE DEFINED FOR BEHAVIORAL ASSESSMENTS. MORRIS WATER MAZE WAS USED TO EVALUATE SPATIAL MEMORY, SHUTTLE BOX WAS USED TO INVESTIGATE AVOIDANCE MEMORY AND A PENTYLENETETRAZOLE-INDUCED SEIZURE WAS USED TO EVALUATE SEIZURE SUSCEPTIBILITY IN THE OFFSPRING. RESULTS: SPATIAL LEARNING AND AVOIDANCE MEMORY SIGNIFICANTLY DECREASED IN BOTH MALE AND FEMALE OFFSPRING OF PG LPS-EXPOSED FEMALE RATS, COMPARED TO THE CONTROL OFFSPRING. LATENCY TO REACH SEIZURE STAGES 1 AND 2 SIGNIFICANTLY INCREASED IN THE MALE OFFSPRING, BUT NOT THE FEMALE OFFSPRING OF PG LPS-EXPOSED FEMALE, COMPARED TO THE CONTROL OFFSPRING. HOWEVER, NO SIGNIFICANT DIFFERENCE WAS FOUND IN LATENCY TO REACH STAGES 3-5. CONCLUSION: PRE-PREGNANCY EXPOSURE TO PG LPS COULD AFFECT SOME BEHAVIORAL FUNCTIONS IN BOTH MALE AND FEMALE OFFSPRING INTERGENERATIONALLY.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
6    66  33 A KEY ROLE FOR EZH2 IN EPIGENETIC SILENCING OF HOX GENES IN MANTLE CELL LYMPHOMA. THE CHROMATIN MODIFIER EZH2 IS OVEREXPRESSED AND ASSOCIATED WITH INFERIOR OUTCOME IN MANTLE CELL LYMPHOMA (MCL). RECENTLY, WE DEMONSTRATED PREFERENTIAL DNA METHYLATION OF HOX GENES IN MCL COMPARED WITH CHRONIC LYMPHOCYTIC LEUKEMIA (CLL), DESPITE THESE GENES NOT BEING EXPRESSED IN EITHER ENTITY. SINCE EZH2 HAS BEEN SHOWN TO REGULATE HOX GENE EXPRESSION, TO GAIN FURTHER INSIGHT INTO ITS POSSIBLE ROLE IN DIFFERENTIAL SILENCING OF HOX GENES IN MCL VS. CLL, WE PERFORMED DETAILED EPIGENETIC CHARACTERIZATION USING REPRESENTATIVE CELL LINES AND PRIMARY SAMPLES. WE OBSERVED SIGNIFICANT OVEREXPRESSION OF EZH2 IN MCL VS. CLL. CHROMATIN IMMUNE PRECIPITATION (CHIP) ASSAYS REVEALED THAT EZH2 CATALYZED REPRESSIVE H3 LYSINE 27 TRIMETHYLATION (H3K27ME3), WHICH WAS SUFFICIENT TO SILENCE HOX GENES IN CLL, WHEREAS IN MCL H3K27ME3 IS ACCOMPANIED BY DNA METHYLATION FOR A MORE STABLE REPRESSION. MORE IMPORTANTLY, HYPERMETHYLATION OF THE HOX GENES IN MCL RESULTED FROM EZH2 OVEREXPRESSION AND SUBSEQUENT RECRUITMENT OF THE DNA METHYLATION MACHINERY ONTO HOX GENE PROMOTERS. THE IMPORTANCE OF EZH2 UPREGULATION IN THIS PROCESS WAS FURTHER UNDERSCORED BY SIRNA TRANSFECTION AND EZH2 INHIBITOR EXPERIMENTS. ALTOGETHER, THESE OBSERVATIONS IMPLICATE EZH2 IN THE LONG-TERM SILENCING OF HOX GENES IN MCL, AND ALLUDE TO ITS POTENTIAL AS A THERAPEUTIC TARGET WITH CLINICAL IMPACT.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
7  2120  32 EPIGENETIC HISTONE MODIFICATIONS IN A CLINICALLY RELEVANT RAT MODEL OF CHRONIC ETHANOL-BINGE-MEDIATED LIVER INJURY. PURPOSE: ETHANOL BINGE AUGMENTS LIVER INJURY AFTER CHRONIC ETHANOL CONSUMPTION IN HUMANS, BUT THE MECHANISM BEHIND THE ENHANCED LIVER INJURY BY ETHANOL BINGE IS NOT KNOWN. IN THIS STUDY WE USED A CLINICALLY RELEVANT RAT MODEL IN WHICH LIVER INJURY IS AMPLIFIED BY BINGE AFTER CHRONIC ETHANOL TREATMENT AND INVESTIGATED THE IMPORTANCE OF HISTONE MODIFICATIONS. METHODS: EIGHT-WEEK-OLD SPRAGUE-DAWLEY RATS WERE FED ETHANOL IN A LIQUID DIET FOR 4 WEEKS. CONTROL RATS WERE FED AN ISOCALORIC LIQUID DIET. THIS WAS FOLLOWED BY THREE BINGE ADMINISTRATIONS OF ETHANOL (INTRAGASTRIC 5 G/KG BODY WEIGHT, 12 H APART). IN THE CONTROL, ETHANOL WAS REPLACED BY WATER. FOUR HOURS AFTER THE LAST BINGE ADMINISTRATION, LIVER SAMPLES WERE ANALYZED FOR HISTONE MODIFICATIONS AND PARAMETERS OF LIVER INJURY. RESULTS: CHRONIC ETHANOL ADMINISTRATION ALONE CAUSED AN INCREASE IN HISTONE H3 SER10 AND SER28 (H3S10 OR S28) PHOSPHORYLATION, AND BINGE ETHANOL REDUCED THEIR LEVELS. LEVELS OF DUALLY MODIFIED PHOSPHOACETYLATED HISTONE H3 (H3ACK9/PS10) INCREASED AFTER ACUTE BINGE ETHANOL AND REMAINED SAME AFTER CHRONIC ETHANOL BINGE. IN CONTRAST, HISTONE H3 LYSINE-9 ACETYLATION (H3ACK9) WAS NOT INCREASED AFTER CHRONIC ETHANOL BUT INCREASED SIGNIFICANTLY AFTER ACUTE BINGE AND CHRONIC ETHANOL BINGE. INCREASE IN HISTONE ACETYLATION WAS ACCOMPANIED BY INCREASED PHOSPHO-ERK1/2 IN THE NUCLEAR EXTRACTS. INCREASED ACETYLATION AFTER CHRONIC ETHANOL BINGE WAS ALSO ACCOMPANIED BY INCREASED PROTEIN LEVELS OF GCN5 HISTONE ACETYL TRANSFERASE AND A MODEST INCREASE IN HDAC3 IN THE NUCLEUS. HISTONE LYSINE-9 DIMETHYLATION WAS SIGNIFICANTLY INCREASED AFTER CHRONIC ETHANOL BINGE. CHRONIC ETHANOL BINGE ALSO RESULTED IN A DECREASE IN THE SAM:SAH RATIO WITH A RELATIVE DECREASE OF SAM LEVELS AND A CORRESPONDING INCREASE IN SAH LEVELS. CONCLUSIONS: ETHANOL BINGE AFTER CHRONIC ETHANOL ALTERED THE PROFILE OF SITE-SPECIFIC HISTONE MODIFICATIONS AND MAY UNDERLIE THE MECHANISM OF AUGMENTED LIVER INJURY BY CHRONIC-ETHANOL-BINGE-TREATED RATS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
8  3619  35 IN VIVO ACUTE ON CHRONIC ETHANOL EFFECTS IN LIVER: A MOUSE MODEL EXHIBITING EXACERBATED INJURY, ALTERED METABOLIC AND EPIGENETIC RESPONSES. CHRONIC ALCOHOLICS WHO ALSO BINGE DRINK (I.E., ACUTE ON CHRONIC) ARE PRONE TO AN EXACERBATED LIVER INJURY BUT ITS MECHANISM IS NOT UNDERSTOOD. WE THEREFORE INVESTIGATED THE IN VIVO EFFECTS OF CHRONIC AND BINGE ETHANOL INGESTION AND COMPARED TO CHRONIC ETHANOL FOLLOWED BY THREE REPEAT BINGE ETHANOL ON THE LIVER OF MALE C57/BL6 MICE FED ETHANOL IN LIQUID DIET (4%) FOR FOUR WEEKS FOLLOWED BY BINGE ETHANOL (INTRAGASTRIC ADMINISTRATION, 3.5 G/KG BODY WEIGHT, THREE DOSES, 12H APART). CHRONIC FOLLOWED BY BINGE ETHANOL EXACERBATED FAT ACCUMULATION, NECROSIS, DECREASE IN HEPATIC SAM AND SAM:SAH RATIO, INCREASE IN ADENOSINE LEVELS, AND ELEVATED CYP2E1 LEVELS. HISTONE H3 LYSINE ACETYLATION (H3ACK9), DUALLY MODIFIED PHOSPHOACETYLATED HISTONE H3 (H3ACK9/PS10), AND PHOSPHORYLATED H2AX INCREASED AFTER BINGE WHEREAS PHOSPHORYLATION OF HISTONE H3 SER 10 (H3S10) AND H3 SER 28 (H3S28) INCREASED AFTER CHRONIC ETHANOL-BINGE. HISTONE H3 LYSINE 4 AND 9 DIMETHYLATION INCREASED WITH A MARKED DIMETHYLATION IN H3K9 IN CHRONIC ETHANOL BINGE GROUP. TRIMETHYLATED HISTONE H3 LEVELS DID NOT CHANGE. NUCLEAR LEVELS OF HISTONE ACETYL TRANSFERASE GCN5 AND HISTONE DEACETYLASE HDAC3 WERE ELEVATED WHEREAS PHOSPHO-CREB DECREASED IN A DISTINCTIVE MANNER. TAKEN TOGETHER, ACUTE ON CHRONIC ETHANOL INGESTION CAUSED AMPLIFICATION OF LIVER INJURY AND ELICITED CHARACTERISTIC PROFILES OF HISTONE MODIFICATIONS, METABOLIC ALTERATIONS, AND CHANGES IN NUCLEAR PROTEIN LEVELS. THESE FINDINGS DEMONSTRATE THAT CHRONIC ETHANOL EXPOSURE RENDERS LIVER MORE SUSCEPTIBLE TO REPEAT ACUTE/BINGE ETHANOL INDUCED ACCELERATION OF ALCOHOLIC LIVER DISEASE.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
9   286  47 AGING AND ALCOHOL INTERACT TO ALTER HEPATIC DNA HYDROXYMETHYLATION. BACKGROUND: AGING AND CHRONIC ALCOHOL CONSUMPTION ARE BOTH MODIFIERS OF DNA METHYLATION, BUT IT IS NOT YET KNOWN WHETHER CHRONIC ALCOHOL CONSUMPTION ALSO ALTERS DNA HYDROXYMETHYLATION, A NEWLY DISCOVERED EPIGENETIC MARK PRODUCED BY OXIDATION OF METHYLCYTOSINE. FURTHERMORE, IT HAS NOT BEEN TESTED WHETHER AGING AND ALCOHOL INTERACT TO MODIFY THIS EPIGENETIC PHENOMENON, THEREBY HAVING AN INDEPENDENT EFFECT ON GENE EXPRESSION. METHODS: OLD (18 MONTHS) AND YOUNG (4 MONTHS) MALE C57BL/6 MICE WERE PAIR-FED EITHER A LIEBER-DECARLI LIQUID DIET WITH ALCOHOL (18% OF ENERGY) OR AN ISOCALORIC LIEBER-DECARLI CONTROL DIET FOR 5 WEEKS. GLOBAL DNA HYDROXYMETHYLATION AND DNA METHYLATION WERE ANALYZED FROM HEPATIC DNA USING A NEW LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY METHOD. HEPATIC MRNA EXPRESSION OF THE TET ENZYMES WERE MEASURED VIA QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION. RESULTS: IN YOUNG MICE, MILD CHRONIC ALCOHOL EXPOSURE SIGNIFICANTLY REDUCED GLOBAL DNA HYDROXYMETHYLATION COMPARED WITH CONTROL MICE (0.22 +/- 0.01 VS. 0.29 +/- 0.06%, P = 0.004). ALCOHOL DID NOT SIGNIFICANTLY ALTER HYDROXYMETHYLCYTOSINE LEVELS IN OLD MICE. OLD MICE FED THE CONTROL DIET SHOWED DECREASED GLOBAL DNA HYDROXYMETHYLATION COMPARED WITH YOUNG MICE FED THE CONTROL DIET (0.24 +/- 0.02 VS. 0.29 +/- 0.06%, P = 0.04). THIS MODEL SUGGESTS AN INTERACTION BETWEEN AGING AND ALCOHOL IN DETERMINING DNA HYDROXYMETHYLATION (PINTERACTION = 0.009). EXPRESSION OF TET2 AND TET3 WAS DECREASED IN THE OLD MICE RELATIVE TO THE YOUNG (P < 0.005). CONCLUSIONS: THE OBSERVATION THAT ALCOHOL ALTERS DNA HYDROXYMETHYLATION INDICATES A NEW EPIGENETIC EFFECT OF ALCOHOL. THIS IS THE FIRST STUDY DEMONSTRATING THE INTERACTIVE EFFECTS OF CHRONIC ALCOHOL CONSUMPTION AND AGING ON DNA HYDROXYMETHYLATION.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
10  5791  41 STABLE HISTONE METHYLATION CHANGES AT PROTEOGLYCAN NETWORK GENES FOLLOWING ETHANOL EXPOSURE. ALCOHOL USE DISORDER (AUD) IS A CHRONIC MENTAL ILLNESS IN WHICH PATIENTS OFTEN ACHIEVE PROTRACTED PERIODS OF ABSTINENCE PRIOR TO RELAPSE. EPIGENETIC MECHANISMS MAY PROVIDE AN EXPLANATION FOR THE PERSISTING GENE EXPRESSION CHANGES THAT CAN BE OBSERVED EVEN AFTER LONG PERIODS OF ABSTINENCE AND MAY CONTRIBUTE TO RELAPSE. IN THIS STUDY, WE EXAMINED TWO HISTONE MODIFICATIONS, HISTONE 3 LYSINE 4 TRI-METHYLATION (H3K4ME3) AND HISTONE 3 LYSINE 27 TRI-METHYLATION (H3K27ME3), IN THE PREFRONTAL CORTEX OF WITHDRAWAL SEIZURE RESISTANT (WSR) MICE 21 DAYS AFTER 72 H OF ETHANOL VAPOR EXPOSURE. THESE HISTONE MODIFICATIONS WERE SELECTED BECAUSE THEY ARE ASSOCIATED WITH ACTIVE PROMOTERS (H3K4ME3) AND REPRESSED GENE EXPRESSION IN A EUCHROMATIC ENVIRONMENT (H3K27ME3). WE PERFORMED A GENOME-WIDE ANALYSIS TO IDENTIFY DIFFERENCES IN H3K4ME3 AND H3K27ME3 LEVELS IN POST-ETHANOL EXPOSURE VS. CONTROL MICE BY CHIP-SEQ. WE DETECTED A GLOBAL REDUCTION IN H3K4ME3 PEAKS AND INCREASE IN H3K27ME3 PEAKS IN POST-ETHANOL EXPOSURE MICE COMPARED TO CONTROLS, THESE CHANGES ARE CONSISTENT WITH PERSISTENT REDUCTIONS IN GENE EXPRESSION. PATHWAY ANALYSIS OF GENES DISPLAYING CHANGES IN H3K4ME3 AND H3K27ME3 REVEALED ENRICHMENT FOR GENES INVOLVED IN PROTEOGLYCAN AND CALCIUM SIGNALING PATHWAYS, RESPECTIVELY. MICROARRAY ANALYSIS OF 7,683 GENES AND QPCR ANALYSIS IDENTIFIED EIGHT GENES DISPLAYING CONCORDANT REGULATION OF GENE EXPRESSION AND H3K4ME3/H3K27ME3. WE ALSO COMPARED CHANGES IN H3K4ME3 AND/OR H3K27ME3 FROM OUR STUDY WITH CHANGES IN GENE EXPRESSION IN RESPONSE TO ETHANOL FROM PUBLISHED LITERATURE AND WE FOUND THAT THE EXPRESSION OF 52% OF THE GENES WITH ALTERED H3K4ME3 BINDING AND 40% OF GENES WITH H3K27ME3 DIFFERENCES ARE ALTERED BY ETHANOL EXPOSURE. THE CHROMATIN CHANGES ASSOCIATED WITH THE 21-DAY POST-EXPOSURE PERIOD SUGGEST THAT THIS PERIOD IS A UNIQUE STATE IN THE ADDICTION CYCLE THAT DIFFERS FROM ETHANOL INTOXICATION AND ACUTE WITHDRAWAL. THESE RESULTS PROVIDE INSIGHTS INTO THE ENDURING EFFECTS OF ETHANOL ON PROTEOGLYCAN AND CALCIUM SIGNALING GENES IN THE BRAIN.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
11  3953  33 LOCUS-SPECIFIC REVERSIBLE DNA METHYLATION REGULATES TRANSIENT IL-10 EXPRESSION IN TH1 CELLS. IL-10 IS A PLEIOTROPIC CYTOKINE WITH MULTIFACETED FUNCTIONS IN ESTABLISHING IMMUNE HOMEOSTASIS. ALTHOUGH EXPRESSED BY TH1 AND TH2 CELLS, CONVENTIONAL TH1 CELLS PRODUCE MARGINAL LEVELS OF IL-10 COMPARED WITH THEIR TH2 COUNTERPARTS. IN THIS STUDY, WE INVESTIGATED THE EPIGENETIC MECHANISMS OF IL-10 GENE EXPRESSION IN TH1 CELLS. BIOINFORMATICS EMBOSS CPG PLOT ANALYSIS AND BISULFITE PYROSEQUENCING REVEALED THREE CPG DNA METHYLATION SITES IN THE IL-10 GENE LOCUS. PROGRESSIVE DNA METHYLATION AT ALL OF THE CPG REGIONS OF INTEREST (ROIS) ESTABLISHED A REPRESSIVE PROGRAM OF IL-10 GENE EXPRESSION IN TH1 CELLS. INTERESTINGLY, TH1 CELLS TREATED WITH IL-12 AND IL-27 CYTOKINES, THEREBY MIMICKING A CHRONIC INFLAMMATORY CONDITION IN VIVO, DISPLAYED A SIGNIFICANT INCREASE IN IL-10 PRODUCTION THAT WAS ACCOMPANIED BY SELECTIVE DNA DEMETHYLATION AT ROI 3 LOCATED IN INTRON 3. IL-10-PRODUCING T CELLS ISOLATED FROM LYMPHOCYTIC CHORIOMENINGITIS VIRUS-INFECTED MICE ALSO SHOWED ENHANCED DNA DEMETHYLATION AT ROI 3. BINDING OF STAT1 AND STAT3 TO DEMETHYLATED ROI 3 ENHANCED IL-10 EXPRESSION IN AN IL-12/IL-27-DEPENDENT MANNER. ACCORDINGLY, CD4(+) T CELLS ISOLATED FROM STAT1- OR STAT3-KNOCKOUT MICE WERE SIGNIFICANTLY DEFECTIVE IN IL-10 PRODUCTION. OUR DATA SUGGEST THAT, ALTHOUGH STABLY MAINTAINED DNA METHYLATION AT THE PROMOTER MAY REPRESS IL-10 EXPRESSION IN TH1 CELLS, LOCUS-SPECIFIC REVERSIBLE DNA DEMETHYLATION MAY SERVE AS A THRESHOLD PLATFORM TO CONTROL TRANSIENT IL-10 GENE EXPRESSION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
12  3722  39 INHIBITION OF DNA METHYLATION DURING CHRONIC OBSTRUCTIVE BLADDER DISEASE (COBD) IMPROVES FUNCTION, PATHOLOGY AND EXPRESSION. PARTIAL BLADDER OUTLET OBSTRUCTION DUE TO PROSTATE HYPERPLASIA OR POSTERIOR URETHRAL VALVES, IS A WIDESPREAD CAUSE OF URINARY DYSFUNCTION, PATIENT DISCOMFORT AND ALSO RESPONSIBLE FOR IMMENSE HEALTH CARE COSTS. EVEN AFTER REMOVAL OR RELIEF OF OBSTRUCTION, THE FUNCTIONAL AND PATHOLOGIC ASPECTS OF OBSTRUCTION REMAIN AS A CHRONIC OBSTRUCTIVE BLADDER DISEASE (COBD). EPIGENETIC CHANGES, SUCH AS DNA METHYLATION, CONTRIBUTE TO THE PERSISTENT CHARACTER OF MANY CHRONIC DISEASES, AND MAY BE ALTERED IN COBD. WE TESTED WHETHER CANDIDATE GENES AND PATHWAYS AND THE PATHOPHYSIOLOGY OF COBD WERE AFFECTED BY A HYPOMETHYLATING AGENT, DECITABINE (DAC). COBD WAS CREATED IN FEMALE SPRAGUE-DAWLEY RATS BY SURGICAL LIGATION OF THE URETHRA FOR 6 WEEKS, FOLLOWED BY REMOVAL OF THE SUTURE. SHAM LIGATIONS WERE PERFORMED BY PASSING THE SUTURE BEHIND THE URETHRA. AFTER REMOVAL OF THE OBSTRUCTION OR SHAM REMOVAL, ANIMALS WERE RANDOMIZED TO DAC TREATMENT (1 MG/KG/3-TIMES/WEEK INTRAPERITONEALLY) OR VEHICLE (NORMAL SALINE). BLADDER FUNCTION WAS NON-INVASIVELY TESTED USING METABOLIC CAGES, BOTH ONE DAY PRIOR TO DE-OBSTRUCTION AT 6 WEEKS AND PRIOR TO SACRIFICE AT 10 WEEKS. RESIDUAL VOLUME AND BLADDER MASS WERE MEASURED FOR EACH BLADDER. BLADDERS WERE EXAMINED BY IMMUNOSTAINING AS WELL AS QPCR. THE EFFECTS OF DNA METHYLTRANSFERASE (DNMT)-3A KNOCKOUT OR OVEREXPRESSION ON SMOOTH MUSCLE CELL (SMC) FUNCTION AND PHENOTYPE WERE ALSO EXAMINED IN BLADDER SMC AND EX VIVO CULTURE. RESIDUAL VOLUMES OF THE DAC TREATED GROUP WERE NOT SIGNIFICANTLY DIFFERENT FROM THE NS GROUP. COMPARED TO COBD NS, COBD DAC TREATMENT HELPED PRESERVE MICTURITION VOLUME WITH A SIGNIFICANT RECOVERY OF THE VOIDING EFFICIENCY (RATIO OF THE MAXIMUM VOIDED VOLUME/MAXIMUM BLADDER CAPACITY) BY ONE THIRD (FIG. 1, P > 0.05). BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) VARIANTS 1 AND 5 WERE UPREGULATED BY COBD AND SIGNIFICANTLY REDUCED BY DAC TREATMENT. DEPOSITION OF COLLAGEN IN THE COBD BLADDER WAS REDUCED BY DAC, BUT GROSS HYPERTROPHY REMAINED. IN BLADDER SMC, DNMT3A OVEREXPRESSION LED TO A LOSS OF CONTRACTILE FUNCTION AND PHENOTYPE. IN BLADDERS, PERSISTENTLY ALTERED BY COBD, INHIBITION OF DNA-METHYLATION ENHANCES FUNCTIONAL RECOVERY, UNLIKE TREATMENT DURING PARTIAL OBSTRUCTION, WHICH EXACERBATES OBSTRUCTIVE PATHOLOGY. THE UNDERLYING MECHANISMS MAY RELATE TO THE GENE EXPRESSION CHANGES IN BDNF AND THEIR EFFECTS ON SIGNALING IN THE BLADDER.	2021	
                                                                                                                            
13  1161  48 CONTINUOUS DEVELOPMENTAL AND EARLY LIFE TRICHLOROETHYLENE EXPOSURE PROMOTED DNA METHYLATION ALTERATIONS IN POLYCOMB PROTEIN BINDING SITES IN EFFECTOR/MEMORY CD4(+) T CELLS. TRICHLOROETHYLENE (TCE) IS AN INDUSTRIAL SOLVENT AND DRINKING WATER POLLUTANT ASSOCIATED WITH CD4(+) T CELL-MEDIATED AUTOIMMUNITY. IN OUR MOUSE MODEL, DISCONTINUATION OF TCE EXPOSURE DURING ADULTHOOD AFTER DEVELOPMENTAL EXPOSURE DID NOT PREVENT IMMUNOTOXICITY. TO DETERMINE WHETHER PERSISTENT EFFECTS WERE LINKED TO EPIGENETIC CHANGES WE CONDUCTED WHOLE GENOME REDUCED REPRESENTATION BISULFITE SEQUENCING (RRBS) TO EVALUATE METHYLATION OF CPG SITES IN AUTOSOMAL CHROMOSOMES IN ACTIVATED EFFECTOR/MEMORY CD4(+) T CELLS. FEMALE MRL+/+ MICE WERE EXPOSED TO VEHICLE CONTROL OR TCE IN THE DRINKING WATER FROM GESTATION UNTIL ~37 WEEKS OF AGE [POSTNATAL DAY (PND) 259]. IN A SUBSET OF MICE, TCE EXPOSURE WAS DISCONTINUED AT ~22 WEEKS OF AGE (PND 154). AT PND 259, RRBS ASSESSMENT REVEALED MORE GLOBAL METHYLATION CHANGES IN THE CONTINUOUS EXPOSURE GROUP VS. THE DISCONTINUOUS EXPOSURE GROUP. A MAJORITY OF THE DIFFERENTIALLY METHYLATED CPG REGIONS (DMRS) ACROSS PROMOTERS, ISLANDS, AND REGULATORY ELEMENTS WERE HYPERMETHYLATED (~90%). HOWEVER, CONTINUOUS DEVELOPMENTAL TCE EXPOSURE ALTERED THE METHYLATION OF 274 CPG SITES IN PROMOTERS AND CPG ISLANDS. IN CONTRAST, ONLY 4 CPG ISLAND REGIONS WERE DIFFERENTIALLY METHYLATED (HYPERMETHYLATED) IN THE DISCONTINUOUS GROUP. INTERESTINGLY, 2 OF THESE 4 SITES WERE ALSO HYPERMETHYLATED IN THE CONTINUOUS EXPOSURE GROUP, AND BOTH OF THESE ISLAND REGIONS ARE ASSOCIATED WITH LYSINE 27 ON HISTONE H3 (H3K27) INVOLVED IN POLYCOMB COMPLEX-DEPENDENT TRANSCRIPTIONAL REPRESSION VIA H3K27 TRI-METHYLATION. CPG SITES WERE OVERLAPPED WITH THE OPEN REGULATORY ANNOTATION DATABASE. UNLIKE THE DISCONTINUOUS GROUP, CONTINUOUS TCE TREATMENT RESULTED IN 129 DMRS INCLUDING 12 UNIQUE TRANSCRIPTION FACTORS AND REGULATORY ELEMENTS; 80% OF WHICH WERE ENRICHED FOR ONE OR MORE POLYCOMB GROUP (PCG) PROTEIN BINDING REGIONS (I.E., SUZ12, EZH2, JARID2, AND MTF2). PATHWAY ANALYSIS OF THE DMRS INDICATED THAT TCE PRIMARILY ALTERED THE METHYLATION OF GENES ASSOCIATED WITH REGULATION OF CELLULAR METABOLISM AND CELL SIGNALING. THE RESULTS DEMONSTRATED THAT CONTINUOUS DEVELOPMENTAL EXPOSURE TO TCE DIFFERENTIALLY METHYLATED BINDING SITES OF PCG PROTEINS IN EFFECTOR/MEMORY CD4(+) CELLS. THERE WERE MINIMAL YET POTENTIALLY BIOLOGICALLY SIGNIFICANT EFFECTS THAT OCCURRED WHEN EXPOSURE WAS DISCONTINUED. THESE RESULTS POINT TOWARD A NOVEL MECHANISM BY WHICH CHRONIC DEVELOPMENTAL TCE EXPOSURE MAY ALTER TERMINALLY DIFFERENTIATED CD4(+) T CELL FUNCTION IN ADULTHOOD.	2019	

14  2347  32 EPIGENETIC REGULATION OF MIR-124 UNDER ETHANOL DEPENDENCE AND WITHDRAWAL. WITHDRAWAL FROM CHRONIC ALCOHOL CAUSE THE PERSISTENT MOLECULAR ALTERATION, SUCH AS CHANGES IN THE RELEASE OF NEUROTRANSMITTER AND GENE EXPRESSION. THE ALTERATIONS ARE THOUGHT TO INCREASE IN THE RISK OF RELAPSE. RECENT STUDIES SUGGEST THAT THE GENE EXPRESSION REGULATED BY HISTONE ACETYLATION MAY PLAY AN IMPORTANT ROLE IN THE DEPENDENCE OF ABUSED DRUGS, INCLUDING OF ETHANOL. FURTHERMORE, MIRNA, ANOTHER REGULATOR OF GENE EXPRESSION, ARE ALSO IMPORTANT MOLECULES FOR THE DEPENDENCE. HOWEVER, CHANGES IN THE MOLECULES UNDER ETHANOL WITHDRAWAL AND ITS RELATIONSHIP ARE POORLY UNDERSTOOD. IN THE PRESENT STUDY, WE INVESTIGATED THE EXPRESSION OF ACETYLATED HISTONE H3 AND MIR-124 IN MOUSE BRAIN AT 3 DAYS AFTER ETHANOL WITHDRAWAL. 6-WEEK AGES OF C57BL/6J MICE WERE TREATED WITH LIQUID DIET CONTAINING ETHANOL FOR 10 DAYS. USING THE ESCALATING ETHANOL DOSAGE SCHEDULE, THE MICE WERE FED THE ETHANOL DIET AS FOLLOWS: 1ST DAY: 1 W/V%: 2ND AND 3RD DAY: 3 W/V%; 4TH AND 5TH DAY: 4 W/V% AND FROM THE 6TH TO 10TH DAY: 5 W/V% ETHANOL DIET, RESPECTIVELY. THE PAIR-FED CONTROL MICE WERE GIVEN THE SAME VOLUME OF ETHANOL-FREE LIQUID DIET WITH GLUCOSE SUBSTITUTED IN ISOCALORIC QUANTITIES FOR ETHANOL. AFTER FEEDING ALCOHOL LIQUID DIET, THE MICE SHOWED SEVERE WITHDRAWAL SIGNS. THE EXPRESSION OF ACETYLATED HISTONE H3 WAS SIGNIFICANTLY DECREASED IN LIMBIC FOREBRAIN AT 3 DAYS AFTER WITHDRAWAL. WE FOUND THAT MIR-124 ALSO DECREASED IN THE LIMBIC FOREBRAIN. IT HAS BEEN REPORTED THAT CDC42 REGULATES NEURONAL DEVELOPMENT AS A TARGET OF MIR-124. WE FOUND THAT CDC42 PROTEIN MARKEDLY INCREASED IN BOTH BRAIN REGIONS AT 3 DAYS AFTER WITHDRAWAL. OUR FINDINGS SUGGEST THAT CHANGES IN THE EXPRESSION OF MIR-124 VIA HISTONE ACETYLATION LEADS TO CHANGE THE CDC42 EXPRESSION UNDER ETHANOL WITHDRAWAL.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
15  4567  45 MYELOID-DERIVED SUPPRESSOR CELL FUNCTION AND EPIGENETIC EXPRESSION EVOLVES OVER TIME AFTER SURGICAL SEPSIS. BACKGROUND: SEPSIS IS AN INCREASINGLY SIGNIFICANT CHALLENGE THROUGHOUT THE WORLD AS ONE OF THE MAJOR CAUSES OF PATIENT MORBIDITY AND MORTALITY. CENTRAL TO THE HOST IMMUNOLOGIC RESPONSE TO SEPSIS IS THE INCREASE IN CIRCULATING MYELOID-DERIVED SUPPRESSOR CELLS (MDSCS), WHICH HAVE BEEN DEMONSTRATED TO BE PRESENT AND INDEPENDENTLY ASSOCIATED WITH POOR LONG-TERM CLINICAL OUTCOMES. MDSCS ARE PLASTIC CELLS AND POTENTIALLY MODIFIABLE, PARTICULARLY THROUGH EPIGENETIC INTERVENTIONS. THE OBJECTIVE OF THIS STUDY WAS TO DETERMINE HOW THE SUPPRESSIVE PHENOTYPE OF MDSCS EVOLVES AFTER SEPSIS IN SURGICAL ICU PATIENTS, AS WELL AS TO IDENTIFY EPIGENETIC DIFFERENCES IN MDSCS THAT MAY EXPLAIN THESE CHANGES. METHODS: CIRCULATING MDSCS FROM 267 SURVIVORS OF SURGICAL SEPSIS WERE PHENOTYPED AT VARIOUS INTERVALS OVER 6 WEEKS, AND HIGHLY ENRICHED MDSCS FROM 23 OF THESE SAMPLES WERE CO-CULTURED WITH CD3/CD28-STIMULATED AUTOLOGOUS T CELLS. MICRORNA EXPRESSION FROM ENRICHED MDSCS WAS ALSO IDENTIFIED. RESULTS: WE OBSERVED THAT MDSC NUMBERS REMAIN SIGNIFICANTLY ELEVATED IN HOSPITALIZED SEPSIS SURVIVORS FOR AT LEAST 6 WEEKS AFTER THEIR INFECTION. HOWEVER, ONLY MDSCS OBTAINED AT AND BEYOND 14 DAYS POST-SEPSIS SIGNIFICANTLY SUPPRESSED T LYMPHOCYTE PROLIFERATION AND IL-2 PRODUCTION. THESE SAME MDSCS DISPLAYED UNIQUE EPIGENETIC (MIRNA) EXPRESSION PATTERNS COMPARED TO EARLIER TIME POINTS. CONCLUSIONS: WE CONCLUDE THAT IN SEPSIS SURVIVORS, IMMATURE MYELOID CELL NUMBERS ARE INCREASED BUT THE IMMUNE SUPPRESSIVE FUNCTION SPECIFIC TO MDSCS DEVELOPS OVER TIME, AND THIS IS ASSOCIATED WITH A SPECIFIC EPIGENOME. THESE FINDINGS MAY EXPLAIN THE CHRONIC AND PERSISTENT IMMUNE SUPPRESSION SEEN IN THESE SUBJECTS.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
16  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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
17  3940  41 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
18  3410  43 HOXA5 UNDERGOES DYNAMIC DNA METHYLATION AND TRANSCRIPTIONAL REPRESSION IN THE ADIPOSE TISSUE OF MICE EXPOSED TO HIGH-FAT DIET. BACKGROUND/OBJECTIVES: THE GENOMIC BASES OF THE ADIPOSE TISSUE ABNORMALITIES INDUCED BY CHRONIC POSITIVE CALORIE EXCESS HAVE BEEN ONLY PARTIALLY ELUCIDATED. WE ADOPTED A GENOME-WIDE APPROACH TO DIRECTLY TEST WHETHER LONG-TERM HIGH-FAT DIET (HFD) EXPOSURE AFFECTS THE DNA METHYLATION PROFILE OF THE MOUSE ADIPOSE TISSUE AND TO IDENTIFY THE FUNCTIONAL CONSEQUENCES OF THESE CHANGES. SUBJECTS/METHODS: WE HAVE USED EPIDIDYMAL FAT OF MICE FED EITHER HIGH-FAT (HFD) OR REGULAR CHOW (STD) DIET FOR 5 MONTHS AND PERFORMED GENOME-WIDE DNA METHYLATION ANALYSES BY METHYLATED DNA IMMUNOPRECIPITATION SEQUENCING (MEDIP-SEQ). MOUSE HOMEOBOX (HOX) GENE DNA METHYLATION PCR, RT-QPCR AND BISULPHITE SEQUENCING ANALYSES WERE THEN PERFORMED. RESULTS: MICE FED THE HFD PROGRESSIVELY EXPANDED THEIR ADIPOSE MASS ACCOMPANIED BY A SIGNIFICANT DECREASE IN GLUCOSE TOLERANCE (P<0.001) AND INSULIN SENSITIVITY (P<0.05). MEDIP-SEQ DATA ANALYSIS REVEALED A UNIFORM DISTRIBUTION OF DIFFERENTIALLY METHYLATED REGIONS (DMR) THROUGH THE ENTIRE ADIPOCYTE GENOME, WITH A HIGHER NUMBER OF HYPERMETHYLATED REGIONS IN HFD MICE (P<0.005). THIS DIFFERENT METHYLATION PROFILE WAS ACCOMPANIED BY INCREASED EXPRESSION OF THE DNMT3A DNA METHYLTRANSFERASE (DNMT; P<0.05) AND THE METHYL-CPG-BINDING DOMAIN PROTEIN MBD3 (P<0.05) GENES IN HFD MICE. GENE ONTOLOGY ANALYSIS REVEALED THAT, IN THE HFD-TREATED MICE, THE HOX FAMILY OF DEVELOPMENT GENES WAS HIGHLY ENRICHED IN DIFFERENTIALLY METHYLATED GENES (P=0.008). TO VALIDATE THIS FINDING, HOXA5, WHICH IS IMPLICATED IN FAT TISSUE DIFFERENTIATION AND REMODELING, HAS BEEN SELECTED AND ANALYZED BY BISULPHITE SEQUENCING, CONFIRMING HYPERMETHYLATION IN THE ADIPOSE TISSUE FROM THE HFD MICE. HOXA5 HYPERMETHYLATION WAS ASSOCIATED WITH DOWNREGULATION OF HOXA5 MRNA AND PROTEIN EXPRESSION. FEEDING ANIMALS PREVIOUSLY EXPOSED TO THE HFD WITH A STANDARD CHOW DIET FOR TWO FURTHER MONTHS IMPROVED THE METABOLIC PHENOTYPE OF THE ANIMALS, ACCOMPANIED BY RETURN OF HOXA5 METHYLATION AND EXPRESSION LEVELS (P<0.05) TO VALUES SIMILAR TO THOSE OF THE CONTROL MICE MAINTAINED UNDER STANDARD CHOW. CONCLUSIONS: HFD INDUCES ADIPOSE TISSUE ABNORMALITIES ACCOMPANIED BY EPIGENETIC CHANGES AT THE HOXA5 ADIPOSE TISSUE REMODELING GENE.	2016	
                                                                                                                                                                                                                                                                                                              
19   223  30 ACUTE PSYCHOSOCIAL STRESS-MEDIATED CHANGES IN THE EXPRESSION AND METHYLATION OF PERFORIN IN CHRONIC FATIGUE SYNDROME. PERFORIN (PRF1) IS ESSENTIAL FOR IMMUNE SURVEILLANCE AND STUDIES REPORT DECREASED PERFORIN IN CHRONIC FATIGUE SYNDROME (CFS), AN ILLNESS POTENTIALLY ASSOCIATED WITH STRESS AND/OR INFECTION. WE HYPOTHESIZE THAT STRESS CAN INFLUENCE REGULATION OF PRF1 EXPRESSION, AND THAT THIS REGULATION WILL DIFFER BETWEEN CFS AND NON-FATIGUED (NF) CONTROLS. WE USED THE TRIER SOCIAL STRESS TEST (TSST) AS A STANDARDIZED ACUTE PSYCHOSOCIAL STRESS, AND EVALUATED ITS EFFECT ON PRF1 EXPRESSION AND METHYLATION IN CFS (N = 34) COMPARED WITH NF (N = 47) PARTICIPANTS. DURING THE TSST, NATURAL KILLER (NK) CELLS INCREASED SIGNIFICANTLY IN BOTH CFS (P = <0.0001) AND NF SUBJECTS (P = <0.0001). UNLIKE PREVIOUS REPORTS, THERE WAS NO SIGNIFICANT DIFFERENCE IN PRF1 EXPRESSION AT BASELINE OR DURING TSST BETWEEN CFS AND NF. HOWEVER, WHOLE BLOOD PRF1 EXPRESSION INCREASED 1.6 FOLD DURING THE TSST IN BOTH CFS (P = 0.0003) AND NF (P = <0.0001). FURTHER, THE PEAK RESPONSE IMMEDIATELY FOLLOWING THE TSST WAS LOWER IN CFS COMPARED WITH NF (P = 0.04). IN ADDITION, AT 1.5 HOURS POST TSST, PRF1 EXPRESSION WAS ELEVATED IN CFS COMPARED WITH NF (WHOLE BLOOD, P = 0.06; PBMC, P = 0.02). METHYLATION OF SEVEN CPG SITES IN THE METHYLATION SENSITIVE REGION OF THE PRF1 PROMOTER RANGED FROM 38%-79% WITH NO SIGNIFICANT DIFFERENCES BETWEEN CFS AND NF. ALTHOUGH, THE AVERAGE BASELINE METHYLATION OF ALL SEVEN CPG SITES DID NOT DIFFER BETWEEN CFS AND NF GROUPS, IT SHOWED A SIGNIFICANT NEGATIVE CORRELATION WITH PRF1 EXPRESSION AT ALL TSST TIME POINTS IN BOTH CFS (R = -0.56, P = <0.0001) AND NF (R = -0.38, P = <0.0001). AMONG PARTICIPANTS WITH HIGH AVERAGE METHYLATION (>/=65%), PRF1 EXPRESSION WAS SIGNIFICANTLY LOWER IN CFS THAN NF SUBJECTS IMMEDIATELY FOLLOWING TSST. THESE FINDINGS SUGGEST METHYLATION COULD BE AN IMPORTANT EPIGENETIC DETERMINANT OF INTER-INDIVIDUAL DIFFERENCES IN PRF1 EXPRESSION AND THAT THE DIFFERENCES IN PRF1 EXPRESSION AND METHYLATION BETWEEN CFS AND NF IN THE ACUTE STRESS RESPONSE REQUIRE FURTHER INVESTIGATION.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
20  5609  31 S-ADENOSYLMETHIONINE DECREASES THE PEAK BLOOD ALCOHOL LEVELS 3 H AFTER AN ACUTE BOLUS OF ETHANOL BY INDUCING ALCOHOL METABOLIZING ENZYMES IN THE LIVER. INTRODUCTION: AN ALCOHOL BOLUS CAUSES THE BLOOD ALCOHOL LEVEL (BAL) TO PEAK AT 1-2 H POST INGESTION. THE ETHANOL ELIMINATION RATE IS REGULATED BY ALCOHOL METABOLIZING ENZYMES, PRIMARILY ALCOHOL DEHYDROGENASE (ADH1), ACETALDEHYDE DEHYDROGENASE (ALDH), AND CYTOCHROME P450 (CYP2E1). RECENTLY, S-ADENOSYLMETHIONINE (SAME) WAS FOUND TO REDUCE ACUTE BALS 3 H AFTER AN ALCOHOL BOLUS. THE QUESTION, THEN, WAS: WHAT IS THE MECHANISM INVOLVED IN THIS REDUCTION OF BAL BY FEEDING SAME? TO ANSWER THIS QUESTION, WE INVESTIGATED THE CHANGES IN ETHANOL METABOLIZING ENZYMES AND THE EPIGENETIC CHANGES THAT REGULATE THE EXPRESSION OF THESE ENZYMES DURING ACUTE BINGE DRINKING AND CHRONIC DRINKING. METHODS: RATS WERE FED A BOLUS OF ETHANOL WITH OR WITHOUT SAME, AND WERE SACRIFICED AT 3 H OR 12 H AFTER THE BOLUS. RESULTS: RT-PCR AND WESTERN BLOT ANALYSES SHOWED THAT SAME SIGNIFICANTLY INDUCED ADH1 LEVELS IN THE 3 H LIVER SAMPLES. HOWEVER, SAME DID NOT AFFECT THE CHANGES IN ADH1 PROTEIN LEVELS 12 H POST BOLUS. SINCE SAME IS A METHYL DONOR, IT WAS POSTULATED THAT THE ADH1 GENE EXPRESSION UP REGULATION AT 3 H WAS DUE TO A HISTONE MODIFICATION INDUCED BY METHYLATION FROM METHYL TRANSFERASES. DIMETHYLATED HISTONE 3 LYSINE 4 (H3K4ME2), A MODIFICATION RESPONSIBLE FOR GENE EXPRESSION ACTIVATION, WAS FOUND TO BE SIGNIFICANTLY INCREASED BY SAME AT 3 H POST BOLUS. CONCLUSION: THESE RESULTS CORRELATED WITH THE LOW BAL FOUND AT 3 H POST BOLUS, AND SUPPORT THE CONCEPT THAT SAME INCREASED THE GENE EXPRESSION TO INCREASE THE ELIMINATION RATE OF ETHANOL IN BINGE DRINKING BY INCREASING H3K4ME2.	2010