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  3808  47 INTRANASAL ADMINISTRATION OF ACINETOBACTER LWOFFII IN A MURINE MODEL OF ASTHMA INDUCES IL-6-MEDIATED PROTECTION ASSOCIATED WITH CECAL MICROBIOTA CHANGES. BACKGROUND: EARLY-LIFE EXPOSURE TO CERTAIN ENVIRONMENTAL BACTERIA INCLUDING ACINETOBACTER LWOFFII (AL) HAS BEEN IMPLICATED IN PROTECTION FROM CHRONIC INFLAMMATORY DISEASES INCLUDING ASTHMA LATER IN LIFE. HOWEVER, THE UNDERLYING MECHANISMS AT THE IMMUNE-MICROBE INTERFACE REMAIN LARGELY UNKNOWN. METHODS: THE EFFECTS OF REPEATED INTRANASAL AL EXPOSURE ON LOCAL AND SYSTEMIC INNATE IMMUNE RESPONSES WERE INVESTIGATED IN WILD-TYPE AND IL6(-/-) , IL10(-/-) , AND IL17(-/-) MICE EXPOSED TO OVALBUMIN-INDUCED ALLERGIC AIRWAY INFLAMMATION. THOSE INVESTIGATIONS WERE EXPANDED BY MICROBIOME ANALYSES. TO ASSESS FOR AL-ASSOCIATED CHANGES IN GENE EXPRESSION, THE PICTURE ARISING FROM ANIMAL DATA WAS SUPPLEMENTED BY IN VITRO EXPERIMENTS OF MACROPHAGE AND T-CELL RESPONSES, YIELDING EXPRESSION AND EPIGENETIC DATA. RESULTS: THE ASTHMA PREVENTIVE EFFECT OF AL WAS CONFIRMED IN THE LUNG. REPEATED INTRANASAL AL ADMINISTRATION TRIGGERED A PROINFLAMMATORY IMMUNE RESPONSE PARTICULARLY CHARACTERIZED BY ELEVATED LEVELS OF IL-6, AND CONSEQUENTLY, IL-6 INDUCED IL-10 PRODUCTION IN CD4(+) T-CELLS. BOTH IL-6 AND IL-10, BUT NOT IL-17, WERE REQUIRED FOR ASTHMA PROTECTION. AL HAD A PROFOUND IMPACT ON THE GENE REGULATORY LANDSCAPE OF CD4(+) T-CELLS WHICH COULD BE LARGELY RECAPITULATED BY RECOMBINANT IL-6. AL ADMINISTRATION ALSO INDUCED MARKED CHANGES IN THE GASTROINTESTINAL MICROBIOME BUT NOT IN THE LUNG MICROBIOME. BY COMPARING THE EFFECTS ON THE MICROBIOTA ACCORDING TO MOUSE GENOTYPE AND AL-TREATMENT STATUS, WE HAVE IDENTIFIED MICROBIAL TAXA THAT WERE ASSOCIATED WITH EITHER DISEASE PROTECTION OR ACTIVITY. CONCLUSION: THESE EXPERIMENTS PROVIDE A NOVEL MECHANISM OF ACINETOBACTER LWOFFII-INDUCED ASTHMA PROTECTION OPERATING THROUGH IL-6-MEDIATED EPIGENETIC ACTIVATION OF IL-10 PRODUCTION AND WITH ASSOCIATED EFFECTS ON THE INTESTINAL MICROBIOME.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
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  4291  37 MICRORNA PROFILES IN INTESTINAL EPITHELIAL CELLS IN A MOUSE MODEL OF SEPSIS. SEPSIS IS A SYSTEMIC INFLAMMATORY DISORDER THAT LEADS TO THE DYSFUNCTION OF MULTIPLE ORGANS. IN THE INTESTINE, THE DEREGULATION OF THE EPITHELIAL BARRIER CONTRIBUTES TO THE DEVELOPMENT OF SEPSIS BY TRIGGERING CONTINUOUS EXPOSURE TO HARMFUL FACTORS. HOWEVER, SEPSIS-INDUCED EPIGENETIC CHANGES IN GENE-REGULATION NETWORKS WITHIN INTESTINAL EPITHELIAL CELLS (IECS) REMAIN UNEXPLORED. IN THIS STUDY, WE ANALYZED THE EXPRESSION PROFILE OF MICRORNAS (MIRNAS) IN IECS ISOLATED FROM A MOUSE MODEL OF SEPSIS GENERATED VIA CECAL SLURRY INJECTION. AMONG 239 MIRNAS, 14 MIRNAS WERE UPREGULATED, AND 9 MIRNAS WERE DOWNREGULATED IN THE IECS BY SEPSIS. UPREGULATED MIRNAS IN IECS FROM SEPTIC MICE, PARTICULARLY MIR-149-5P, MIR-466Q, MIR-495, AND MIR-511-3P, WERE SEEN TO EXHIBIT COMPLEX AND GLOBAL EFFECTS ON GENE REGULATION NETWORKS. INTERESTINGLY, MIR-511-3P HAS EMERGED AS A DIAGNOSTIC MARKER IN THIS SEPSIS MODEL DUE TO ITS INCREASE IN BLOOD IN ADDITION TO IECS. AS EXPECTED, MRNAS IN THE IECS WERE REMARKABLY ALTERED BY SEPSIS; SPECIFICALLY, 2248 MRNAS WERE DECREASED, WHILE 612 MRNAS WERE INCREASED. THIS QUANTITATIVE BIAS MAY BE POSSIBLY DERIVED, AT LEAST PARTLY, FROM THE DIRECT EFFECTS OF THE SEPSIS-INCREASED MIRNAS ON THE COMPREHENSIVE EXPRESSION OF MRNAS. THUS, CURRENT IN SILICO DATA INDICATE THAT THERE ARE DYNAMIC REGULATORY RESPONSES OF MIRNAS TO SEPSIS IN IECS. IN ADDITION, THE MIRNAS THAT WERE INCREASED WITH SEPSIS HAD ENRICHED DOWNSTREAM PATHWAYS INCLUDING WNT SIGNALING, WHICH IS ASSOCIATED WITH WOUND HEALING, AND FGF/FGFR SIGNALING, WHICH HAS BEEN LINKED TO CHRONIC INFLAMMATION AND FIBROSIS. THESE MODIFICATIONS IN MIRNA NETWORKS IN IECS MAY LEAD TO BOTH PRO- AND ANTI-INFLAMMATORY EFFECTS IN SEPSIS. THE FOUR MIRNAS DISCOVERED ABOVE WERE SHOWN TO PUTATIVELY TARGET LOX, PTCH1, COL22A1, FOXO1, OR HMGA2, VIA IN SILICO ANALYSIS, WHICH WERE ASSOCIATED WITH WNT OR INFLAMMATORY PATHWAYS AND SELECTED FOR FURTHER STUDY. THE EXPRESSIONS OF THESE TARGET GENES WERE DOWNREGULATED IN SEPSIS IECS, POSSIBLY THROUGH POSTTRANSCRIPTIONAL MODIFICATIONS OF THESE MIRNAS. TAKEN TOGETHER, OUR STUDY SUGGESTS THAT IECS DISPLAY A DISTINCTIVE MIRNA PROFILE WHICH IS CAPABLE OF COMPREHENSIVELY AND FUNCTIONALLY RESHAPING THE IEC-SPECIFIC MRNA LANDSCAPE IN A SEPSIS MODEL.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
5  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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
6  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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
7   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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
8  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	
                                                                                                                                                                                                                                                                     
9  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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
10   912  31 CHRONIC EXPOSURE TO WATER POLLUTANT TRICHLOROETHYLENE INCREASED EPIGENETIC DRIFT IN CD4(+) T CELLS. AIM: AUTOIMMUNE DISEASE AND CD4(+) T-CELL ALTERATIONS ARE INDUCED IN MICE EXPOSED TO THE WATER POLLUTANT TRICHLOROETHYLENE (TCE). WE EXAMINED HERE WHETHER TCE ALTERED GENE-SPECIFIC DNA METHYLATION IN CD4(+) T CELLS AS A POSSIBLE MECHANISM OF IMMUNOTOXICITY. MATERIALS & METHODS: NAIVE AND EFFECTOR/MEMORY CD4(+) T CELLS FROM MICE EXPOSED TO TCE (0.5 MG/ML IN DRINKING WATER) FOR 40 WEEKS WERE EXAMINED BY BISULFITE NEXT-GENERATION DNA SEQUENCING. RESULTS: A PROBABILISTIC MODEL CALCULATED FROM MULTIPLE GENES SHOWED THAT TCE DECREASED METHYLATION CONTROL IN CD4(+) T CELLS. DATA FROM INDIVIDUAL GENES FITTED TO A QUADRATIC REGRESSION MODEL SHOWED THAT TCE INCREASED GENE-SPECIFIC METHYLATION VARIANCE IN BOTH CD4 SUBSETS. CONCLUSION: TCE INCREASED EPIGENETIC DRIFT OF SPECIFIC CPG SITES IN CD4(+) T CELLS.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
11  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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
12  5760  46 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
13  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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
14  6382  30 THE ROLE OF PARTICULATE MATTERS ON METHYLATION OF IFN-GAMMA AND IL-4 PROMOTER GENES IN PEDIATRIC ALLERGIC RHINITIS. ALLERGIC RHINITIS (AR) IS A CHRONIC INFLAMMATORY DISORDER DRIVEN BY T CELL ACTIVATION. HOW PARTICULATE MATTER CONTRIBUTES TO EPIGENETIC CHANGES THAT IN TURN INFLUENCE CYTOKINE GENE EXPRESSION IN CD4(+)T CELLS REMAINS UNCLEAR. IN THIS STUDY, 105 CHILDREN DIAGNOSED WITH AR AND 90 HEALTHY CONTROLS WERE RECRUITED TO EXPLORE THE POSSIBLE MECHANISM OF PARTICULATE MATTER (PM) ON THE EPIGENETIC REGULATION OF CD4(+)T IFN-GAMMA AND IL-4 PROMOTER GENES. DAILY AVERAGE PM(10) AND PM(2.5) WERE OBTAINED FROM FIVE STATE-CONTROLLED MONITORING STATIONS, AND ACTIVITY-BASED DYNAMIC EXPOSURE AND PERSONAL EXPOSURE DATA WERE COLLECTED. DNA METHYLATION PATTERNS OF IFN-GAMMA AND IL-4 PROMOTER REGIONS WERE ANALYZED USING BISULFITE SEQUENCING. MRNA LEVELS WERE DETECTED BY REAL-TIME QUANTITATIVE REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION. WE FOUND THAT THE METHYLATION RATE IN IFN-GAMMA WAS HIGHER IN AR CD4(+)T CELLS THAN IN THE CONTROLS. IFN-GAMMA MRNA EXPRESSION WAS SIGNIFICANTLY DECREASED IN CD4(+)T CELLS, AND NEGATIVELY CORRELATED WITH THE MEAN METHYLATION LEVEL OF IFN-GAMMA. HOWEVER, NO CORRELATION BETWEEN IL-4 METHYLATION AND IL-4 MRNA EXPRESSION WAS FOUND. AFTER ADJUSTING FOR AGE, GENDER, EXCLUSIVE BREASTFEEDING WITHIN 4 MONTHS AFTER BIRTH AND PARENTAL HISTORY OF ALLERGIC DISEASE, OUT DATA SHOWED THAT PM(2.5) EXPOSURE LEVEL WAS POSITIVELY CORRELATED WITH METHYLATION LEVEL IN IFN-GAMMA PROMOTER REGION AND DECREASED CYTOKINE EXPRESSION. WE CONCLUDE THAT THE EFFECT OF PM(2.5) ON PEDIATRIC AR MAY BE MEDIATED THROUGH EPIGENETIC MODIFICATION OF IFN-GAMMA PROMOTER REGION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
15   312  35 ALCOHOL FEEDING IN MICE PROMOTES COLONIC HYPERPERMEABILITY AND CHANGES IN COLONIC ORGANOID STEM CELL FATE. BACKGROUND: ALCOHOL INCREASES INTESTINAL PERMEABILITY TO PROINFLAMMATORY MICROBIAL PRODUCTS THAT PROMOTE LIVER DISEASE, EVEN AFTER A PERIOD OF SOBRIETY. WE SOUGHT TO TEST THE HYPOTHESIS THAT ALCOHOL AFFECTS INTESTINAL STEM CELLS USING AN IN VIVO MODEL AND EX VIVO ORGANOIDS GENERATED FROM JEJUNUM AND COLON FROM MICE FED CHRONIC ALCOHOL. METHODS: MICE WERE FED A CONTROL OR AN ALCOHOL DIET. INTESTINAL PERMEABILITY, LIVER STEATOSIS-INFLAMMATION, AND STOOL SHORT-CHAIN FATTY ACIDS (SCFAS) WERE MEASURED. JEJUNUM AND COLONIC ORGANOIDS AND TISSUE WERE STAINED FOR STEM CELL, CELL LINEAGE, AND APICAL JUNCTION MARKERS WITH ASSESSMENT OF MRNA BY PCR AND RNA-SEQ. CHIP-PCR ANALYSIS WAS CARRIED OUT FOR NOTCH1 USING AN ANTIBODY SPECIFIC FOR ACETYLATED HISTONE 3. RESULTS: ALCOHOL-FED MICE EXHIBITED COLONIC (BUT NOT SMALL INTESTINAL) HYPERPERMEABILITY, STEATOHEPATITIS, AND DECREASED BUTYRATE/TOTAL SCFA RATIO IN STOOL. STEM CELL, CELL LINEAGE, AND APICAL JUNCTION MARKER STAINING IN TISSUE OR ORGANOIDS FROM JEJUNUM TISSUE WERE NOT IMPACTED BY ALCOHOL. ONLY CHROMOGRANIN A (CHGA) WAS INCREASED IN JEJUNUM ORGANOIDS BY QPCR. HOWEVER, COLONIC TISSUE AND ORGANOID STAINING EXHIBITED AN ALCOHOL-INDUCED SIGNIFICANT DECREASE IN CYTOKERATIN 20+ (KRT20+) ABSORPTIVE LINEAGE ENTEROCYTES, A DECREASE IN OCCLUDIN AND E-CADHERIN APICAL JUNCTION PROTEINS, AN INCREASE IN CHGA, AND AN INCREASE IN THE LGR5 STEM CELL MARKER. QPCR REVEALED AN ALCOHOL-INDUCED DECREASE IN COLONIC ORGANOID AND TISSUE NOTCH1, HES1, AND KRT20 AND INCREASED CHGA, SUPPORTING AN ALTERATION IN STEM CELL FATE DUE TO DECREASED NOTCH1 EXPRESSION. COLONIC TISSUE CHIP-PCR REVEALED ALCOHOL FEEDING SUPPRESSED NOTCH1 MRNA EXPRESSION (VIA DEACETYLATION OF HISTONE H3) AND DECREASED NOTCH1 TISSUE STAINING. CONCLUSIONS: DATA SUPPORT A MODEL FOR ALCOHOL-INDUCED COLONIC HYPERPERMEABILITY VIA EPIGENETIC EFFECTS ON NOTCH1, AND THUS HES1, SUPPRESSION THROUGH A MECHANISM INVOLVING HISTONE H3 DEACETYLATION AT THE NOTCH1 LOCUS. THIS DECREASED ENTEROCYTE AND INCREASED ENTEROENDOCRINE CELL COLONIC STEM CELL FATE AND DECREASED APICAL JUNCTIONAL PROTEINS LEADING TO HYPERPERMEABILITY.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
16  2374  41 EPIGENETIC REGULATION OF TNFA EXPRESSION IN PERIODONTAL DISEASE. BACKGROUND: TUMOR NECROSIS FACTOR-ALPHA (TNF-ALPHA) PLAYS A CENTRAL ROLE IN THE MOLECULAR PATHOGENESIS OF PERIODONTAL DISEASE. HOWEVER, THE EPIGENETIC REGULATION ATTRIBUTABLE TO MICROBIAL AND INFLAMMATORY SIGNALS AT THE BIOFILM-GINGIVAL INTERFACE ARE POORLY UNDERSTOOD. IN THIS STUDY, THE DNA METHYLATION ALTERATION WITHIN THE TNFA PROMOTER IN HUMAN GINGIVAL BIOPSIES FROM DIFFERENT STAGES OF PERIODONTAL DISEASE IS INVESTIGATED AND THE REGULATORY MECHANISM OF TNFA TRANSCRIPTION BY DNA METHYLATION IS EXPLORED. METHODS: GINGIVAL BIOPSIES WERE OBTAINED FROM 17 PATIENTS WITH CHRONIC PERIODONTITIS (CP) AND 18 PERIODONTALLY HEALTHY INDIVIDUALS. ANOTHER 11 INDIVIDUALS PARTICIPATED IN AN EXPERIMENTALLY INDUCED GINGIVITIS STUDY, AND GINGIVAL BIOPSIES WERE COLLECTED AT THE BASELINE, INDUCTION, AND RESOLUTION PHASE. TO CONFIRM THAT TNFA PROMOTER METHYLATION MODULATED TNFA TRANSCRIPTION, THP.1 CELLS WERE TREATED WITH A DNA METHYLTRANSFERASE INHIBITOR, 5-AZA-2-DEOXYCYTIDINE (5-AZA-2DC), AND AN RAW294.7 CELL LINE TRANSFECTED WITH A TNFA PROMOTER-SPECIFIC LUCIFERASE REPORTER SYSTEM WITH OR WITHOUT METHYLATION WAS USED. RESULTS: IN GINGIVAL BIOPSIES FROM INDIVIDUALS WITH SEVERE CP, TWO INDIVIDUAL CYTOSINE-GUANINE DINUCLEOTIDES (CPG SITES) WITHIN THE TNFA PROMOTER (AT -163 AND -161 BP) DISPLAYED INCREASED METHYLATION IN CP SAMPLES COMPARED TO THOSE WITH GINGIVAL HEALTH (16.1% +/- 5.1% VERSUS 11.0% +/- 4.6%, P = 0.02 AND 19.8% +/- 4.1% VERSUS 15.4% +/- 3.6%, P = 0.04, RESPECTIVELY). THE METHYLATION LEVEL AT -163 BP WAS INVERSELY ASSOCIATED WITH THE TRANSCRIPTION LEVEL OF TNFA (P = 0.018). HOWEVER, NO SIGNIFICANT DIFFERENCE IN THE TNFA PROMOTER METHYLATION PATTERN WAS OBSERVED IN SAMPLES BIOPSIED DURING THE INDUCTION OR RESOLUTION PHASE OF EXPERIMENTALLY INDUCED GINGIVITIS, WHICH REPRESENTED A REVERSIBLE PERIODONTAL LESION. THP.1 CELLS TREATED WITH 5-AZA-2DC DEMONSTRATED A TIME-DEPENDENT INCREASE IN TNFA MESSENGER LEVEL. IT WAS ALSO FOUND THAT THE LUCIFERASE ACTIVITY DECREASED 2.6-FOLD IN A CONSTRUCT CONTAINING AN IN VITRO METHYLATED TNFA PROMOTER WHEN COMPARED TO THE UNMETHYLATED INSERT (P = 0.03). CONCLUSION: ALTHOUGH THE BIOPSY SAMPLES REPRESENTED A MIXED CELL POPULATION, THE CHANGE IN PROMOTER METHYLATION STATUS IN CHRONIC PERIODONTAL DISEASE SUGGESTED THAT DNA METHYLATION MAY BE AN IMPORTANT REGULATORY MECHANISM IN CONTROLLING TNFA TRANSCRIPTIONAL EXPRESSION IN PERIODONTAL DISEASE.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                    
17   910  39 CHRONIC EXPOSURE TO ETHANOL OF MALE MICE BEFORE MATING PRODUCES ATTENTION DEFICIT HYPERACTIVITY DISORDER-LIKE PHENOTYPE ALONG WITH EPIGENETIC DYSREGULATION OF DOPAMINE TRANSPORTER EXPRESSION IN MOUSE OFFSPRING. PRECONCEPTION EXPOSURE TO ETOH THROUGH THE PATERNAL ROUTE MAY AFFECT NEUROBEHAVIORAL AND DEVELOPMENTAL FEATURES OF OFFSPRING. THIS STUDY INVESTIGATES THE EFFECTS OF PATERNAL EXPOSURE TO ETOH BEFORE CONCEPTION ON THE HYPERACTIVITY, INATTENTION, AND IMPULSIVITY BEHAVIOR OF MALE OFFSPRING IN MICE. SIRE MICE WERE TREATED WITH ETOH IN A CONCENTRATION RANGE APPROXIMATING HUMAN BINGE DRINKING (0-4 G/KG/DAY ETOH) FOR 7 WEEKS AND MATED WITH UNTREATED FEMALES MICE TO PRODUCE OFFSPRING. ETOH EXPOSURE TO SIRE MICE INDUCED ATTENTION DEFICIT HYPERACTIVITY DISORDER (ADHD)-LIKE HYPERACTIVE, INATTENTIVE, AND IMPULSIVE BEHAVIORS IN OFFSPRING. AS A MECHANISTIC LINK, BOTH PROTEIN AND MRNA EXPRESSION OF DOPAMINE TRANSPORTER (DAT), A KEY DETERMINANT OF ADHD-LIKE PHENOTYPES IN EXPERIMENTAL ANIMALS AND HUMANS, WERE SIGNIFICANTLY DECREASED BY PATERNAL ETOH EXPOSURE IN CEREBRAL CORTEX AND STRIATUM OF OFFSPRING MICE ALONG WITH INCREASED METHYLATION OF A CPG REGION OF THE DAT GENE PROMOTER. THE INCREASE IN METHYLATION OF DAT GENE PROMOTER WAS ALSO OBSERVED IN THE SPERM OF SIRE MICE, SUGGESTING GERMLINE CHANGES IN THE EPIGENETIC METHYLATION SIGNATURE OF DAT GENE BY ETOH EXPOSURE. IN ADDITION, THE EXPRESSION OF TWO KEY REGULATORS OF METHYLATION-DEPENDENT EPIGENETIC REGULATION OF FUNCTIONAL GENE EXPRESSION, NAMELY, MECP2 AND DNMT1, WAS MARKEDLY DECREASED IN OFFSPRING CORTEX AND STRIATUM SIRED BY ETOH-EXPOSED MICE. THESE RESULTS SUGGEST THAT PRECONCEPTIONAL EXPOSURE TO ETOH THROUGH THE PATERNAL ROUTE INDUCES BEHAVIORAL CHANGES IN OFFSPRING, POSSIBLY VIA EPIGENETIC CHANGES IN GENE EXPRESSION, WHICH IS ESSENTIAL FOR THE REGULATION OF ADHD-LIKE BEHAVIORS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
18  3304  47 HIGH-GLUCOSE CONCENTRATIONS CHANGE DNA METHYLATION LEVELS IN HUMAN IVM OOCYTES. STUDY QUESTION: WHAT ARE THE EFFECTS OF HIGH-GLUCOSE CONCENTRATIONS ON DNA METHYLATION OF HUMAN OOCYTES? SUMMARY ANSWER: HIGH-GLUCOSE CONCENTRATIONS ALTERED DNA METHYLATION LEVELS OF PEG3 AND ADIPONECTIN IN HUMAN IN VITRO MATURATION OOCYTES. WHAT IS KNOWN ALREADY: MATERNAL DIABETES HAS A DETRIMENTAL INFLUENCE ON OOCYTE QUALITY INCLUDING EPIGENETIC MODIFICATIONS, AS SHOWN IN NON-HUMAN MAMMALIAN SPECIES. STUDY DESIGN, SIZE, DURATION: IMMATURE METAPHASE I (MI) STAGE OOCYTES OF GOOD QUALITY WERE RETRIEVED FROM PATIENTS WHO HAD NORMAL OVARIAN POTENTIAL AND WHO UNDERWENT ICSI IN THE REPRODUCTIVE MEDICINE CENTER OF PEOPLE'S HOSPITAL OF ZHENGZHOU UNIVERSITY. MI OOCYTES WERE CULTURED IN MEDIUM WITH DIFFERENT GLUCOSE CONCENTRATIONS (CONTROL, 10 MM AND 15 MM) IN VITRO AND 48 H LATER, OOCYTES WITH FIRST POLAR BODY EXTRUSION WERE COLLECTED TO CHECK THE DNA METHYLATION LEVELS. PARTICIPANTS/MATERIALS, SETTING, METHODS: MI OOCYTES UNDERWENT IN VITRO MATURATION (IVM) AT 37 DEGREES C WITH 5% MIXED GAS FOR 48 H. THEN THE MATURE OOCYTES WERE TREATED WITH BISULFITE BUFFER. TARGET SEQUENCES WERE AMPLIFIED USING NESTED OR HALF-NESTED PCR AND THE DNA METHYLATION STATUS WAS TESTED USING COMBINED BISULFITE RESTRICTION ANALYSIS (COBRA) AND BISULFITE SEQUENCING (BS). MAIN RESULTS AND THE ROLE OF CHANCE: HIGH-GLUCOSE CONCENTRATIONS SIGNIFICANTLY DECREASED THE FIRST POLAR BODY EXTRUSION RATE. COMPARED TO CONTROLS, THE DNA METHYLATION LEVELS OF PEG3 IN HUMAN IVM OOCYTES WERE SIGNIFICANTLY HIGHER IN 10 MM (P < 0.001) AND 15 MM (P < 0.001) CONCENTRATIONS OF GLUCOSE. BUT THE DNA METHYLATION LEVEL OF H19 WAS NOT AFFECTED BY HIGH-GLUCOSE CONCENTRATIONS IN HUMAN IVM OOCYTES. WE ALSO FOUND THAT THERE WAS A DECREASE IN DNA METHYLATION LEVELS IN THE PROMOTER OF ADIPONECTIN IN HUMAN IVM OOCYTES BETWEEN CONTROLS AND OOCYTES EXPOSED TO 10 MM GLUCOSE (P = 0.028). LARGE SCALE DATA: N/A. LIMITATIONS REASONS FOR CAUTION: IT IS NOT CLEAR WHETHER THE ALTERATIONS ARE BENEFICIAL OR NOT FOR THE EMBRYO DEVELOPMENT AND OFFSPRING HEALTH. THE EFFECTS OF HIGH-GLUCOSE CONCENTRATIONS ON THE WHOLE PROCESS OF OOCYTE MATURATION ARE STILL NOT ELUCIDATED. ANOTHER ISSUE IS THAT THE NUMBER OF OOCYTES USED IN THIS STUDY WAS LIMITED. WIDER IMPLICATIONS OF THE FINDINGS: THIS IS THE FIRST TIME THAT THE EFFECTS OF HIGH-GLUCOSE CONCENTRATION ON DNA METHYLATION OF HUMAN OOCYTES HAVE BEEN ELUCIDATED. OUR RESULT INDICATES THAT IN HUMANS, THE HIGH RISK OF CHRONIC DISEASES IN OFFSPRING FROM DIABETIC MOTHERS MAY ORIGINATE FROM ABNORMAL DNA MODIFICATIONS IN OOCYTES. STUDY FUNDING/COMPETING INTEREST(S): THIS WORK WAS SUPPORTED BY THE FUND OF NATIONAL NATURAL SCIENCE FOUNDATION OF CHINA (81401198) AND DOCTOR FOUNDATION OF QINGDAO AGRICULTURAL UNIVERSITY (1116008).THE AUTHORS DECLARE THAT THERE ARE NO POTENTIAL CONFLICTS OF INTEREST RELEVANT TO THIS ARTICLE.	2018	

19  6540  39 TRANSCRIPTIONAL, EPIGENETIC, AND FUNCTIONAL REPROGRAMMING OF MONOCYTES FROM NON-HUMAN PRIMATES FOLLOWING CHRONIC ALCOHOL DRINKING. CHRONIC HEAVY DRINKING (CHD) OF ALCOHOL IS A KNOWN RISK FACTOR FOR INCREASED SUSCEPTIBILITY TO BACTERIAL AND VIRAL INFECTION AS WELL AS IMPAIRED WOUND HEALING. EVIDENCE SUGGESTS THAT THESE DEFECTS ARE MEDIATED BY A DYSREGULATED INFLAMMATORY RESPONSE ORIGINATING FROM MYELOID CELLS, NOTABLY MONOCYTES AND MACROPHAGES, BUT THE MECHANISMS REMAIN POORLY UNDERSTOOD. OUR ABILITY TO STUDY CHD IS IMPACTED BY THE COMPLEXITIES OF HUMAN DRINKING PATTERNS AND BEHAVIOR AS WELL AS COMORBIDITIES AND CONFOUNDING RISK FACTORS FOR PATIENTS WITH ALCOHOL USE DISORDERS. TO OVERCOME THESE CHALLENGES, WE UTILIZED A TRANSLATIONAL RHESUS MACAQUE MODEL OF VOLUNTARY ETHANOL SELF-ADMINISTRATION THAT CLOSELY RECAPITULATES HUMAN DRINKING PATTERNS AND CHRONICITY. IN THIS STUDY, WE EXAMINED THE EFFECTS OF CHD ON BLOOD MONOCYTES IN CONTROL AND CHD FEMALE MACAQUES AFTER 12 MONTHS OF DAILY ETHANOL CONSUMPTION. WHILE MONOCYTES FROM CHD FEMALE MACAQUES GENERATED A HYPER-INFLAMMATORY RESPONSE TO EX VIVO LPS STIMULATION, THEIR RESPONSE TO E. COLI WAS DAMPENED. IN DEPTH SCRNA-SEQ ANALYSIS OF PURIFIED MONOCYTES REVEALED SIGNIFICANT SHIFTS IN CLASSICAL MONOCYTE SUBSETS WITH ACCUMULATION OF CELLS EXPRESSING MARKERS OF HYPOXIA (HIF1A) AND INFLAMMATION (NFKB SIGNALING PATHWAY) IN CHD MACAQUES. THE INCREASED PRESENCE OF MONOCYTE SUBSETS SKEWED TOWARDS INFLAMMATORY PHENOTYPES WAS COMPLEMENTED BY EPIGENETIC ANALYSIS, WHICH REVEALED HIGHER ACCESSIBILITY OF PROMOTER REGIONS THAT REGULATE GENES INVOLVED IN CYTOKINE SIGNALING PATHWAYS. COLLECTIVELY, DATA PRESENTED IN THIS MANUSCRIPT DEMONSTRATE THAT CHD SHIFTS CLASSICAL MONOCYTE SUBSET COMPOSITION AND PRIMES THE MONOCYTES TOWARDS A MORE HYPER-INFLAMMATORY RESPONSE TO LPS, BUT COMPROMISED PATHOGEN RESPONSE.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
20   169  40 ABNORMALITIES OF AMPK ACTIVATION AND GLUCOSE UPTAKE IN CULTURED SKELETAL MUSCLE CELLS FROM INDIVIDUALS WITH CHRONIC FATIGUE SYNDROME. BACKGROUND: POST EXERTIONAL MUSCLE FATIGUE IS A KEY FEATURE IN CHRONIC FATIGUE SYNDROME (CFS). ABNORMALITIES OF SKELETAL MUSCLE FUNCTION HAVE BEEN IDENTIFIED IN SOME BUT NOT ALL PATIENTS WITH CFS. TO TRY TO LIMIT POTENTIAL CONFOUNDERS THAT MIGHT CONTRIBUTE TO THIS CLINICAL HETEROGENEITY, WE DEVELOPED A NOVEL IN VITRO SYSTEM THAT ALLOWS COMPARISON OF AMP KINASE (AMPK) ACTIVATION AND METABOLIC RESPONSES TO EXERCISE IN CULTURED SKELETAL MUSCLE CELLS FROM CFS PATIENTS AND CONTROL SUBJECTS. METHODS: SKELETAL MUSCLE CELL CULTURES WERE ESTABLISHED FROM 10 SUBJECTS WITH CFS AND 7 AGE-MATCHED CONTROLS, SUBJECTED TO ELECTRICAL PULSE STIMULATION (EPS) FOR UP TO 24H AND EXAMINED FOR CHANGES ASSOCIATED WITH EXERCISE. RESULTS: IN THE BASAL STATE, CFS CULTURES SHOWED INCREASED MYOGENIN EXPRESSION BUT DECREASED IL6 SECRETION DURING DIFFERENTIATION COMPARED WITH CONTROL CULTURES. CONTROL CULTURES SUBJECTED TO 16 H EPS SHOWED A SIGNIFICANT INCREASE IN BOTH AMPK PHOSPHORYLATION AND GLUCOSE UPTAKE COMPARED WITH UNSTIMULATED CELLS. IN CONTRAST, CFS CULTURES SHOWED NO INCREASE IN AMPK PHOSPHORYLATION OR GLUCOSE UPTAKE AFTER 16 H EPS. HOWEVER, GLUCOSE UPTAKE REMAINED RESPONSIVE TO INSULIN IN THE CFS CELLS POINTING TO AN EXERCISE-RELATED DEFECT. IL6 SECRETION IN RESPONSE TO EPS WAS SIGNIFICANTLY REDUCED IN CFS COMPARED WITH CONTROL CULTURES AT ALL TIME POINTS MEASURED. CONCLUSION: EPS IS AN EFFECTIVE MODEL FOR ELICITING MUSCLE CONTRACTION AND THE METABOLIC CHANGES ASSOCIATED WITH EXERCISE IN CULTURED SKELETAL MUSCLE CELLS. WE FOUND FOUR MAIN DIFFERENCES IN CULTURED SKELETAL MUSCLE CELLS FROM SUBJECTS WITH CFS; INCREASED MYOGENIN EXPRESSION IN THE BASAL STATE, IMPAIRED ACTIVATION OF AMPK, IMPAIRED STIMULATION OF GLUCOSE UPTAKE AND DIMINISHED RELEASE OF IL6. THE RETENTION OF THESE DIFFERENCES IN CULTURED MUSCLE CELLS FROM CFS SUBJECTS POINTS TO A GENETIC/EPIGENETIC MECHANISM, AND PROVIDES A SYSTEM TO IDENTIFY NOVEL THERAPEUTIC TARGETS.	2015