1  1653 141 DOPAMINE TRANSPORTER KNOCKOUT RATS DISPLAY EPIGENETIC ALTERATIONS IN RESPONSE TO COCAINE EXPOSURE. (1) BACKGROUND: THERE IS AN URGENT NEED FOR EFFECTIVE TREATMENTS FOR COCAINE USE DISORDER (CUD), AND NEW PHARMACOLOGICAL APPROACHES TARGETING EPIGENETIC MECHANISMS APPEAR TO BE PROMISING OPTIONS FOR THE TREATMENT OF THIS DISEASE. DOPAMINE TRANSPORTER (DAT) TRANSGENIC RATS RECENTLY HAVE BEEN PROPOSED AS A NEW ANIMAL MODEL FOR STUDYING SUSCEPTIBILITY TO CUD. (2) METHODS: DAT TRANSGENIC RATS WERE TREATED CHRONICALLY WITH COCAINE (10 MG/KG) FOR 8 DAYS, AND THE EXPRESSION OF EPIGENETIC MODULATORS, LYSINE DEMETHYLASE 6B (KDM6B) AND BROMODOMAIN-CONTAINING PROTEIN 4 (BRD4), WAS EXAMINED IN THE PREFRONTAL CORTEX (PFC). (3) RESULTS: WE SHOW THAT ONLY FULL KNOCKOUT (KO) OF DAT IMPACTS BASAL LEVELS OF KDM6B IN FEMALES. ADDITIONALLY, COCAINE ALTERED THE EXPRESSION OF BOTH EPIGENETIC MARKERS IN A SEX- AND GENOTYPE-DEPENDENT MANNER. IN RESPONSE TO CHRONIC COCAINE, KDM6B EXPRESSION WAS DECREASED IN MALE RATS WITH PARTIAL DAT MUTATION (HET), WHILE NO CHANGES WERE OBSERVED IN WILD-TYPE (WT) OR KO RATS. INDEED, WHILE HET MALE RATS HAVE REDUCED KDM6B AND BRD4 EXPRESSION, HET FEMALE RATS SHOWED INCREASED KDM6B AND BRD4 EXPRESSION LEVELS, HIGHLIGHTING THE IMPACT OF SEX ON EPIGENETIC MECHANISMS IN RESPONSE TO COCAINE. FINALLY, BOTH MALE AND FEMALE KO RATS SHOWED INCREASED EXPRESSION OF BRD4, BUT ONLY KO FEMALES EXHIBITED SIGNIFICANTLY INCREASED KDM6B EXPRESSION IN RESPONSE TO COCAINE. ADDITIONALLY, THE MAGNITUDE OF THESE EFFECTS WAS BIGGER IN FEMALES WHEN COMPARED TO MALES FOR BOTH EPIGENETIC ENZYMES. (4) CONCLUSIONS: THIS PRELIMINARY STUDY PROVIDES ADDITIONAL SUPPORT THAT TARGETING KDM6B AND/OR BRD4 MAY POTENTIALLY BE THERAPEUTIC IN TREATING ADDICTION-RELATED BEHAVIORS IN A SEX-DEPENDENT MANNER.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
2  4603  35 NEGATIVE ALLOSTERIC MODULATION OF MGLUR5 PARTIALLY CORRECTS PATHOPHYSIOLOGY IN A MOUSE MODEL OF RETT SYNDROME. RETT SYNDROME (RTT) IS CAUSED BY MUTATIONS IN THE GENE ENCODING METHYL-CPG BINDING PROTEIN 2 (MECP2), AN EPIGENETIC REGULATOR OF MRNA TRANSCRIPTION. HERE, WE REPORT A TEST OF THE HYPOTHESIS OF SHARED PATHOPHYSIOLOGY OF RTT AND FRAGILE X, ANOTHER MONOGENIC CAUSE OF AUTISM AND INTELLECTUAL DISABILITY. IN FRAGILE X, THE LOSS OF THE MRNA TRANSLATIONAL REPRESSOR FMRP LEADS TO EXAGGERATED PROTEIN SYNTHESIS DOWNSTREAM OF METABOTROPIC GLUTAMATE RECEPTOR 5 (MGLUR5). WE FOUND THAT MGLUR5- AND PROTEIN-SYNTHESIS-DEPENDENT SYNAPTIC PLASTICITY WERE SIMILARLY ALTERED IN AREA CA1 OF MECP2 KO MICE. CA1 PYRAMIDAL CELL-TYPE-SPECIFIC, GENOME-WIDE PROFILING OF RIBOSOME-BOUND MRNAS WAS PERFORMED IN WILD-TYPE AND MECP2 KO HIPPOCAMPAL CA1 NEURONS TO REVEAL THE MECP2-REGULATED "TRANSLATOME." WE FOUND SIGNIFICANT OVERLAP BETWEEN RIBOSOME-BOUND TRANSCRIPTS OVEREXPRESSED IN THE MECP2 KO AND FMRP MRNA TARGETS. THESE TENDED TO ENCODE LONG GENES THAT WERE FUNCTIONALLY RELATED TO EITHER CYTOSKELETON ORGANIZATION OR THE DEVELOPMENT OF NEURONAL CONNECTIVITY. IN THE FMR1 KO MOUSE, CHRONIC TREATMENT WITH MGLUR5-NEGATIVE ALLOSTERIC MODULATORS (NAMS) HAS BEEN SHOWN TO AMELIORATE MANY MUTANT PHENOTYPES BY CORRECTING EXCESSIVE PROTEIN SYNTHESIS. IN MECP2 KO MICE, WE FOUND THAT MGLUR5 NAM TREATMENT SIGNIFICANTLY REDUCED THE LEVEL OF OVEREXPRESSED RIBOSOME-ASSOCIATED TRANSCRIPTS, PARTICULARLY THOSE THAT WERE ALSO FMRP TARGETS. SOME RETT PHENOTYPES WERE ALSO AMELIORATED BY TREATMENT, MOST NOTABLY HIPPOCAMPAL CELL SIZE AND LIFESPAN. TOGETHER, THESE RESULTS SUGGEST A POTENTIAL MECHANISTIC LINK BETWEEN MECP2-MEDIATED TRANSCRIPTION REGULATION AND MGLUR5/FMRP-MEDIATED PROTEIN TRANSLATION REGULATION THROUGH COREGULATION OF A SUBSET OF GENES RELEVANT TO SYNAPTIC FUNCTIONS. SIGNIFICANCE STATEMENT: ALTERED REGULATION OF SYNAPTIC PROTEIN SYNTHESIS HAS BEEN HYPOTHESIZED TO CONTRIBUTE TO THE PATHOPHYSIOLOGY THAT UNDERLIES MULTIPLE FORMS OF INTELLECTUAL DISABILITY AND AUTISM SPECTRUM DISORDER. HERE, WE SHOW IN A MOUSE MODEL OF RETT SYNDROME (MECP2 KO) THAT METABOTROPIC GLUTAMATE RECEPTOR 5 (MGLUR5)- AND PROTEIN-SYNTHESIS-DEPENDENT SYNAPTIC PLASTICITY ARE ABNORMAL IN THE HIPPOCAMPUS. WE FOUND THAT A SUBSET OF RIBOSOME-BOUND MRNAS WAS ABERRANTLY UPREGULATED IN HIPPOCAMPAL CA1 NEURONS OF MECP2 KO MICE, THAT THESE SIGNIFICANTLY OVERLAPPED WITH FMRP DIRECT TARGETS AND/OR SFARI HUMAN AUTISM GENES, AND THAT CHRONIC TREATMENT OF MECP2 KO MICE WITH AN MGLUR5-NEGATIVE ALLOSTERIC MODULATOR TUNES DOWN UPREGULATED RIBOSOME-BOUND MRNAS AND PARTIALLY IMPROVES MUTANT MICE PHENOTYPES.	2016	

3   267  35 AFFECTIVE AND COGNITIVE BEHAVIOR IN THE ALPHA-GALACTOSIDASE A DEFICIENT MOUSE MODEL OF FABRY DISEASE. FABRY DISEASE IS AN X-LINKED INHERITED LYSOSOMAL STORAGE DISORDER WITH INTRACELLULAR ACCUMULATION OF GLOBOTRIAOSYLCERAMIDE (GB3) DUE TO ALPHA-GALACTOSIDASE A (ALPHA-GAL A) DEFICIENCY. FABRY PATIENTS FREQUENTLY REPORT OF ANXIETY, DEPRESSION, AND IMPAIRED COGNITIVE FUNCTION. WE CHARACTERIZED AFFECTIVE AND COGNITIVE PHENOTYPE OF MALE MICE WITH ALPHA-GAL A DEFICIENCY (FABRY KO) AND COMPARED RESULTS WITH THOSE OF AGE-MATCHED MALE WILDTYPE (WT) LITTERMATES. YOUNG (3 MONTHS) AND OLD (>/= 18 MONTHS) MICE WERE TESTED IN THE NAIVE STATE AND AFTER I.PL. INJECTION OF COMPLETE FREUND;S ADJUVANT (CFA) AS AN INFLAMMATORY PAIN MODEL. WE USED THE ELEVATED PLUS MAZE (EPM), THE LIGHT-DARK BOX (LDB) AND THE OPEN FIELD TEST (OF) TO INVESTIGATE ANXIETY-LIKE BEHAVIOR. THE FORCED SWIM TEST (FST) AND MORRIS WATER MAZE (MWM) WERE APPLIED TO ASSESS DEPRESSIVE-LIKE AND LEARNING BEHAVIOR. THE EPM TEST REVEALED NO INTERGROUP DIFFERENCE FOR ANXIETY-LIKE BEHAVIOR IN NAIVE YOUNG AND OLD FABRY KO MICE COMPARED TO WT LITTERMATES, EXCEPT FOR LONGER TIME SPENT IN OPEN ARMS OF THE EPM FOR YOUNG WT MICE COMPARED TO YOUNG FABRY KO MICE (P<0.05). AFTER CFA INJECTION, YOUNG FABRY KO MICE SHOWED INCREASED ANXIETY-LIKE BEHAVIOR COMPARED TO YOUNG WT LITTERMATES (P<0.05) AND NAIVE YOUNG FABRY KO MICE (P<0.05) IN THE EPM AS REFLECTED BY SHORTER TIME SPENT IN EPM OPEN ARMS. THERE WERE NO RELEVANT DIFFERENCES IN THE LDB AND THE OF TEST, EXCEPT FOR LONGER TIME SPENT IN THE CENTER ZONE OF THE OF BY YOUNG WT MICE COMPARED TO YOUNG FABRY KO MICE (P<0.05). COMPLEMENTARY TO THIS, DEPRESSION-LIKE AND LEARNING BEHAVIOR WERE NOT DIFFERENT BETWEEN GENOTYPES AND AGE-GROUPS, EXCEPT FOR THE EXPECTEDLY LOWER MEMORY PERFORMANCE IN OLDER AGE-GROUPS COMPARED TO YOUNG MICE. OUR RESULTS INDICATE THAT GENETIC INFLUENCES ON AFFECTIVE AND COGNITIVE SYMPTOMS IN FD MAY BE OF SUBORDINATE RELEVANCE, DRAWING ATTENTION TO POTENTIAL INFLUENCES OF ENVIRONMENTAL AND EPIGENETIC FACTORS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
4  3584  35 IMPACT OF SOCIAL DEFEAT STRESS ON DNA METHYLATION IN DRD2, NR3C1, AND STMN1 IN WILD-TYPE AND STMN1 KNOCK-OUT MICE. OBJECTIVE: EPIGENETIC PROFILES CAN BE MODIFIED BY STRESS. DOPAMINE RECEPTOR D2 (DRD2), GLUCOCORTICOID RECEPTOR GENE (NR3C1) AND STATHMIN 1 (STMN1) GENES ARE ALL IMPLICATED IN ADAPTATION TO STRESS. THE AIM OF STUDY IS TO INVESTIGATE IMPACT OF SOCIAL DEFEAT ON DNA METHYLATION IN DRD2, NR3C1, AND STMN1 IN WILD-TYPE (WT) AND STMN1 KNOCK-OUT (KO) MICE. METHODS: THE WT AND STMN1 KO MICE WERE SUBJECTED TO CHRONIC SOCIAL DEFEAT. BRAIN TISSUES OF THE PREFRONTAL CORTEX (PFC), AMYGDALA (AMY) AND HIPPOCAMPUS (HIP) WERE OBTAINED. WE MEASURED DNA METHYLATION LEVELS OF THE DRD2, NR3C1, AND STMN1 GENES IN THE PFC, AMY, AND HIP USING PYROSEQUENCING. RESULTS: IN WT MICE, SOCIAL DEFEAT STRESS DID NOT INDUCE ANY CHANGES IN DRD2 METHYLATION, WHEREAS SIGNIFICANT HYPERMETHYLATION OCCURRED IN NR3C1 AND STMN1 IN THE SUSCEPTIBLE AND UNSUSCEPTIBLE GROUPS, RESPECTIVELY, COMPARED TO THE CONTROL GROUP. THE METHYLATION RESPONSES IN THE STMN1 KO MICE DIFFERED FROM THOSE SEEN IN THE WT MICE, SUCH THAT HYPERMETHYLATION WAS EVIDENT IN ALL THREE GENES IN THE SUSCEPTIBLE AND UNSUSCEPTIBLE GROUPS COMPARED TO CONTROL GROUP. COMPARISON OF THE STMN1 KO AND WT MICE REVEALED THE SAME PATTERN OF HYPERMETHYLATION FOR ALL THREE GENES. CONCLUSION: SOCIAL DEFEAT STRESS INDUCED DIFFERENT EPIGENETIC MODIFICATIONS IN THREE GENES AMONG CONTROL, UNSUSCEPTIBLE, AND SUSCEPTIBLE GROUPS OF WT AND STMN1 KO MICE. IN PARTICULAR, HYPERMETHYLATION OF NR3C1 IN THE HIP OF THE SUSCEPTIBLE GROUP, AND OF STMN1 IN THE AMY OF THE UNSUSCEPTIBLE GROUP IN WT MICE, COULD SERVE AS EPIGENETIC BIOMARKERS OF STRESS SUSCEPTIBILITY AND STRESS RESILIENCE, RESPECTIVELY.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
5   423  38 ANRIL REGULATES MULTIPLE MOLECULES OF PATHOGENETIC SIGNIFICANCE IN DIABETIC NEPHROPATHY. BACKGROUND: HYPERGLYCEMIA-INDUCED TRANSCRIPTIONAL ALTERATIONS LEAD TO ABERRANT SYNTHESIS OF A LARGE NUMBER OF PATHOGENETIC MOLECULES LEADING TO FUNCTIONAL AND STRUCTURAL DAMAGE TO MULTIPLE END ORGANS INCLUDING THE KIDNEYS. DIABETIC NEPHROPATHY (DN) REMAINS A MAJOR CAUSE OF END STAGE RENAL DISEASE. MULTIPLE EPIGENETIC MECHANISMS, INCLUDING ALTERATION OF LONG NON-CODING RNAS (LNCRNAS) MAY PLAY A SIGNIFICANT ROLE MEDIATING THE CELLULAR TRANSCRIPTIONAL ACTIVITIES. WE HAVE PREVIOUSLY SHOWN THAT LNCRNA ANRIL MAY MEDIATE DIABETES ASSOCIATED MOLECULAR, FUNCTIONAL AND STRUCTURAL ABNORMALITIES IN DN. HERE WE EXPLORED DOWNSTREAM MECHANISMS OF ANRIL ALTERATION IN DN. METHODS: WE USED RENAL CORTICAL TISSUES FROM ANRIL KNOCKOUT (KO) MICE AND WILD TYPE (WT) MICE, WITH OR WITHOUT STREPTOZOTOCIN (STZ) INDUCED DIABETES FOR RNA SEQUENCING. THE DIFFERENTIALLY EXPRESSED GENES WERE IDENTIFIED USING EDGER AND DESEQ2 COMPUTATIONAL METHODS. KEGG AND REACTOME PATHWAY ANALYSES AND NETWORK ANALYSES USING STRING AND IPA WERE SUBSEQUENTLY PERFORMED. RESULTS: DIABETIC ANIMALS SHOWED HYPERGLYCEMIA, REDUCED BODY WEIGHT GAIN, POLYURIA AND INCREASED URINARY ALBUMIN. BOTH ALBUMINURIA AND POLYURIA WERE CORRECTED IN THE KO DIABETIC MICE. RNA ANALYSES SHOWED DIABETES INDUCED ALTERATIONS OF A LARGE NUMBER OF TRANSCRIPTS IN THE WILD TYPE (WT) ANIMALS. ANRIL KNOCKOUT (KO) PREVENTED A LARGE NUMBER OF SUCH ALTERATIONS. THE ALTERED TRANSCRIPTS INCLUDE METABOLIC PATHWAYS, APOPTOSIS, EXTRACELLULAR MATRIX PROTEIN SYNTHESIS AND DEGRADATION, NFKB RELATED PATHWAYS, AGE-RAGE INTERACTION PATHWAYS ETC. ANRIL KO PREVENTED MAJORITY OF THESE PATHWAYS. CONCLUSION: THESE FINDINGS SUGGEST THAT AS ANRIL REGULATES A LARGE NUMBER OF MOLECULES OF PATHOGENETIC SIGNIFICANCE, IT MAY POTENTIALLY BE A DRUG TARGET FOR DN AND OTHER CHRONIC DIABETIC COMPLICATIONS.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
6  5717  38 SIRTUIN-6-DEPENDENT GENETIC AND EPIGENETIC ALTERATIONS ARE ASSOCIATED WITH POOR CLINICAL OUTCOME IN HEPATOCELLULAR CARCINOMA PATIENTS. SIRTUIN 6 (SIRT6) IS A MEMBER OF THE SIRTUIN FAMILY OF NAD+-DEPENDENT DEACETYLASES. GENETIC DELETION OF SIRT6 IN MICE RESULTS IN A SEVERE DEGENERATIVE PHENOTYPE WITH IMPAIRED LIVER FUNCTION AND PREMATURE DEATH. THE ROLE OF SIRT6 IN DEVELOPMENT AND PROGRESSION OF HEPATOCELLULAR CARCINOMA IS CURRENTLY UNKNOWN. WE FIRST INVESTIGATED SIRT6 EXPRESSION IN 153 PRIMARY HUMAN LIVER CANCERS AND IN NORMAL AND CIRRHOTIC LIVERS USING MICROARRAY ANALYSIS. SIRT6 WAS SIGNIFICANTLY DOWN-REGULATED IN BOTH CIRRHOTIC LIVERS AND CANCER. A SIRT6 KNOCKOUT (KO) GENE EXPRESSION SIGNATURE WAS GENERATED FROM PRIMARY HEPATOCTYES ISOLATED FROM 3-WEEK-OLD SIRT6-DEFICIENT ANIMALS. SIRT6-DEFICIENT HEPATOCYTES SHOWED UP-REGULATION OF ESTABLISHED HEPATOCELLULAR CARCINOMA (HCC) BIOMARKERS ALPHA-FETOPROTEIN (AFP), INSULIN-LIKE GROWTH FACTOR 2 (IGF2), H19, AND GLYPICAN-3. FURTHERMORE, DECREASED SIRT6 EXPRESSION WAS OBSERVED IN HEPATOMA CELL LINES THAT ARE KNOWN TO BE APOPTOSIS-INSENSITIVE. RE-EXPRESSION OF SIRT6 IN HEPG2 CELLS INCREASED APOPTOSIS SENSITIVITY TO CD95-STIMULATION OR CHEMOTHERAPY TREATMENT. LOSS OF SIRT6 WAS CHARACTERIZED BY ONCOGENIC CHANGES, SUCH AS GLOBAL HYPOMETHYLATION, AS WELL AS METABOLIC CHANGES, SUCH AS HYPOGLYCEMIA AND INCREASED FAT DEPOSITION. THE HEPATOCYTE-SPECIFIC SIRT6-KO SIGNATURE HAD A PROGNOSTIC IMPACT AND WAS ENRICHED IN PATIENTS WITH POORLY DIFFERENTIATED TUMORS WITH HIGH AFP LEVELS AS WELL AS RECURRENT DISEASE. FINALLY, WE DEMONSTRATED THAT THE SIRT6-KO SIGNATURE POSSESSED A PREDICTIVE VALUE FOR TUMORS OTHER THAN HCC (E.G., BREAST AND LUNG CANCER). CONCLUSION: LOSS OF SIRT6 INDUCES EPIGENETIC CHANGES THAT MAY BE RELEVANT TO CHRONIC LIVER DISEASE AND HCC DEVELOPMENT. DOWN-REGULATION OF SIRT6 AND GENES DYSREGULATED BY LOSS OF SIRT6 POSSESS ONCOGENIC EFFECTS IN HEPATOCARCINOGENESIS. OUR DATA DEMONSTRATE THAT DEFICIENCY IN ONE EPIGENETIC REGULATOR PREDISPOSES A TUMORIGENIC PHENOTYPE THAT ULTIMATELY HAS RELEVANCE FOR OUTCOME OF HCC AND OTHER CANCER PATIENTS.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
7   684  35 BRAIN STAT5 MODULATES LONG-TERM METABOLIC AND EPIGENETIC CHANGES INDUCED BY PREGNANCY AND LACTATION IN FEMALE MICE. SEVERAL METABOLIC AND BEHAVIORAL ADAPTATIONS THAT EMERGE DURING PREGNANCY REMAIN PRESENT AFTER WEANING. THUS, REPRODUCTIVE EXPERIENCE CAUSES LONG-LASTING METABOLIC PROGRAMMING, PARTICULARLY IN THE BRAIN. HOWEVER, THE ISOLATE EFFECTS OF PREGNANCY OR LACTATION AND THE MOLECULAR MECHANISMS INVOLVED IN THESE LONG-TERM MODIFICATIONS ARE CURRENTLY UNKNOWN. IN THE CURRENT STUDY, WE INVESTIGATED THE ROLE OF BRAIN SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION-5 (STAT5), A KEY TRANSCRIPTION FACTOR RECRUITED BY HORMONES HIGHLY SECRETED DURING GESTATION OR LACTATION, FOR THE LONG-TERM ADAPTATIONS INDUCED BY REPRODUCTIVE EXPERIENCE. IN CONTROL MICE, PREGNANCY FOLLOWED BY LACTATION LED TO INCREASED BODY ADIPOSITY AND REDUCED AMBULATORY ACTIVITY LATER IN LIFE. ADDITIONALLY, PREGNANCY+LACTATION INDUCED LONG-TERM EPIGENETIC MODIFICATIONS IN THE BRAIN: WE OBSERVED UPREGULATION IN HYPOTHALAMIC EXPRESSION OF HISTONE DEACETYLASES AND REDUCED NUMBERS OF NEURONS WITH HISTONE H3 ACETYLATION IN THE PARAVENTRICULAR, ARCUATE, AND VENTROMEDIAL NUCLEI. REMARKABLY, BRAIN-SPECIFIC STAT5 ABLATION PREVENTED ALL METABOLIC AND EPIGENETIC CHANGES OBSERVED IN REPRODUCTIVELY EXPERIENCED CONTROL FEMALE MICE. NONETHELESS, BRAIN-SPECIFIC STAT5 KNOCKOUT (KO) MICE THAT HAD THE EXPERIENCE OF PREGNANCY BUT DID NOT LACTATE SHOWED INCREASED BODY WEIGHT AND REDUCED ENERGY EXPENDITURE LATER IN LIFE, WHEREAS PREGNANCY KO AND PREGNANCY+LACTATION KO MICE EXHIBITED IMPROVED INSULIN SENSITIVITY COMPARED WITH VIRGIN KO MICE. IN SUMMARY, LACTATION IS NECESSARY FOR THE LONG-LASTING METABOLIC EFFECTS OBSERVED IN REPRODUCTIVELY EXPERIENCED FEMALE MICE. IN ADDITION, EPIGENETIC MECHANISMS INVOLVING HISTONE ACETYLATION IN NEURONAL POPULATIONS RELATED TO ENERGY BALANCE REGULATION ARE POSSIBLY ASSOCIATED WITH THESE LONG-TERM CONSEQUENCES. FINALLY, OUR FINDINGS HIGHLIGHTED THE KEY ROLE PLAYED BY BRAIN STAT5 SIGNALING FOR THE CHRONIC METABOLIC AND EPIGENETIC CHANGES INDUCED BY PREGNANCY AND LACTATION.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
8  5716  30 SIRT6 PROTECTS VASCULAR SMOOTH MUSCLE CELLS FROM OSTEOGENIC TRANSDIFFERENTIATION VIA RUNX2 IN CHRONIC KIDNEY DISEASE. VASCULAR CALCIFICATION (VC) IS REGARDED AS AN IMPORTANT PATHOLOGICAL CHANGE LACKING EFFECTIVE TREATMENT AND ASSOCIATED WITH HIGH MORTALITY. SIRTUIN 6 (SIRT6) IS A MEMBER OF THE SIRTUIN FAMILY, A CLASS III HISTONE DEACETYLASE AND A KEY EPIGENETIC REGULATOR. SIRT6 HAS A PROTECTIVE ROLE IN PATIENTS WITH CHRONIC KIDNEY DISEASE (CKD). HOWEVER, THE EXACT ROLE AND MOLECULAR MECHANISM OF SIRT6 IN VC IN PATIENTS WITH CKD REMAIN UNCLEAR. HERE, WE DEMONSTRATED THAT SIRT6 WAS MARKEDLY DOWNREGULATED IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) AND IN THE RADIAL ARTERY TISSUE OF PATIENTS WITH CKD WITH VC. SIRT6-TRANSGENIC (SIRT6-TG) MICE SHOWED ALLEVIATED VC, WHILE VASCULAR SMOOTH MUSCLE CELL-SPECIFIC (VSMC-SPECIFIC) SIRT6 KNOCKED-DOWN MICE SHOWED SEVERE VC IN CKD. SIRT6 SUPPRESSED THE OSTEOGENIC TRANSDIFFERENTIATION OF VSMCS VIA REGULATION OF RUNT-RELATED TRANSCRIPTION FACTOR 2 (RUNX2). COIMMUNOPRECIPITATION (CO-IP) AND IMMUNOPRECIPITATION (IP) ASSAYS CONFIRMED THAT SIRT6 BOUND TO RUNX2. MOREOVER, RUNX2 WAS DEACETYLATED BY SIRT6 AND FURTHER PROMOTED NUCLEAR EXPORT VIA EXPORTIN 1 (XPO1), WHICH IN TURN CAUSED DEGRADATION OF RUNX2 THROUGH THE UBIQUITIN-PROTEASOME SYSTEM. THESE RESULTS DEMONSTRATED THAT SIRT6 PREVENTED VC BY SUPPRESSING THE OSTEOGENIC TRANSDIFFERENTIATION OF VSMCS, AND AS SUCH TARGETING SIRT6 MAY BE AN APPEALING THERAPEUTIC TARGET FOR VC IN CKD.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
9  6388  34 THE ROLE OF SIRT1 IN THE BASOLATERAL AMYGDALA IN DEPRESSION-LIKE BEHAVIORS IN MICE. PREVIOUS INVESTIGATIONS HAVE IMPLICATED THE BASOLATERAL AMYGDALA (BLA) EPIGENETIC MECHANISMS IN THE PATHOPHYSIOLOGY OF DEPRESSION. SIRT1 IS A NAD+-DEPENDENT CLASS III HISTONE DEACETYLASE, WIDELY EXPRESSES IN BLA. HOWEVER, EPIGENETIC MECHANISMS IN THE BLA UNDER THE REGULATION OF SIRT1 IN THE DEPRESSION ARE LARGELY UNCHARACTERIZED. UNDER THE CHRONIC UNPREDICTABLE CHRONIC MILD STRESS (CUMS) MOUSE MODEL, WE USED ADENO-ASSOCIATED VIRAL VECTORS (AAV) THAT ENCODED SIRT1-SHRNA OR SIRT1 TO SPECIFICALLY KNOCKDOWN OR OVEREXPRESS SIRT1 IN BLA NEURONS, RESPECTIVELY. CUMS PROCEDURE INDUCED SIGNIFICANT DEPRESSION SYMPTOMS INCLUDING THE DECREASED SUCROSE PREFERENCE, THE LESS BODYWEIGHT GAINED, THE DECREASED IMMOBILE LATENCY AND THE INCREASED IMMOBILE TIME BOTH IN FORCED SWIM TEST (FST) AND TAIL SUSPENSION TEST (TST). KNOCKDOWN OF SIRT1 IN BLA GLUTAMATERGIC NEURONS REVERSED THESE DEPRESSION-LIKE BEHAVIORS AND RESTORED THE SYNAPTIC ABNORMALITIES. OVEREXPRESSION OF SIRT1 IN BLA GLUTAMATERGIC NEURONS INDUCED DEPRESSION-LIKE BEHAVIORS IN NON-STRESSED CONTROL MICE. THE RESULT OF PROTEIN EXPRESSIONS AND ULTRASTRUCTURAL CHANGES WERE CONSISTENT WITH THE BEHAVIORAL RESULTS. OUR STUDY SUGGESTED THAT DOWNREGULATION OF SIRT1 IN BLA HAS CERTAIN BENEFICIAL EFFECT ON CUMS-INDUCED DEPRESSION-LIKE BEHAVIORS SUCH AS ANOREXIA, ANHEDONIA, HOPELESSNESS AND DESPAIR. IN ADDITION, THE INCREASED EXPRESSION OF SIRT1 MAY BE THE IMMEDIATE CAUSE OF DEPRESSIVE-LIKE SYMPTOMS. THE ABNORMAL EXPRESSION OF SIRT1 MAY AFFECT THE TRANSCRIPTIONAL REGULATION MECHANISM AND SIGNALING PROTEIN ACETYLATION, AFFECTING NEUROPLASTICITY AND ULTIMATELY CONTRIBUTE TO MDD. IN THE STRESS-SUSCEPTIBLE MICE, THESE TWO MECHANISMS MAY CO-EXIST, BUT THE SPECIFIC MECHANISM NEEDS FURTHER RESEARCH.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
10  1003  43 CHRONIC TREATMENT WITH FLUOXETINE INDUCES SEX-DEPENDENT ANALGESIC EFFECTS AND MODULATES HDAC2 AND MGLU2 EXPRESSION IN FEMALE MICE. GENDER AND SEX DIFFERENCES IN PAIN RECOGNITION AND DRUG RESPONSES HAVE BEEN REPORTED IN CLINICAL TRIALS AND EXPERIMENTAL MODELS OF PAIN. AMONG ANTIDEPRESSANTS, CONTRADICTORY RESULTS HAVE BEEN OBSERVED IN PATIENTS TREATED WITH SELECTIVE SEROTONIN REUPTAKE INHIBITORS (SSRIS). THIS STUDY EVALUATED SEX DIFFERENCES IN RESPONSE TO THE SSRI FLUOXETINE AFTER CHRONIC ADMINISTRATION IN THE MOUSE FORMALIN TEST. ADULT MALE AND FEMALE CD1 MICE WERE INTRAPERITONEALLY INJECTED WITH FLUOXETINE (10 MG/KG) FOR 21 DAYS AND SUBJECTED TO PAIN ASSESSMENT. FLUOXETINE TREATMENT REDUCED THE SECOND PHASE OF THE FORMALIN TEST ONLY IN FEMALE MICE WITHOUT PRODUCING BEHAVIORAL CHANGES IN MALES. WE ALSO OBSERVED THAT FLUOXETINE WAS ABLE TO SPECIFICALLY INCREASE THE EXPRESSION OF METABOTROPIC GLUTAMATE RECEPTOR TYPE-2 (MGLU2) IN FEMALES. ALSO A REDUCED EXPRESSION OF THE EPIGENETIC MODIFYING ENZYME, HISTONE DEACETYLASE 2 (HDAC2), IN DORSAL ROOT GANGLIA (DRG) AND DORSAL HORN (DH) TOGETHER WITH AN INCREASE HISTONE 3 ACETYLATION (H3) LEVEL WAS OBSERVED IN FEMALES BUT NOT IN MALES. WITH THIS STUDY WE PROVIDE EVIDENCE THAT FLUOXETINE INDUCES SEX SPECIFIC CHANGES IN HDAC2 AND MGLU2 EXPRESSION IN THE DH OF THE SPINAL CORD AND IN DRGS AND SUGGESTS A MOLECULAR EXPLANATION FOR THE ANALGESIC EFFECTS IN FEMALE MICE.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
11  4161  35 MECP2 REGULATES ETHANOL SENSITIVITY AND INTAKE. WE HAVE INVESTIGATED THE EXPRESSION OF CHROMATIN-REGULATING GENES IN THE PREFRONTAL CORTEX AND IN THE SHELL SUBDIVISION OF THE NUCLEUS ACCUMBENS DURING PROTRACTED WITHDRAWAL IN MICE WITH INCREASED ETHANOL DRINKING AFTER CHRONIC INTERMITTENT ETHANOL (CIE) VAPOR EXPOSURE AND IN MICE WITH A HISTORY OF NON-DEPENDENT DRINKING. WE OBSERVED THAT THE METHYL-CPG BINDING PROTEIN 2 (MECP2) WAS ONE OF THE FEW CHROMATIN-REGULATING GENES TO BE DIFFERENTIALLY REGULATED BY A HISTORY OF DEPENDENCE. AS MECP2 HAS THE POTENTIAL OF ACTING AS A BROAD GENE REGULATOR, WE INVESTIGATED SENSITIVITY TO ETHANOL AND ETHANOL DRINKING IN MECP2(308/) (Y) MICE, WHICH HARBOR A TRUNCATED MECP2 ALLELE BUT HAVE A MILDER PHENOTYPE THAN MECP2 NULL MICE. WE OBSERVED THAT MECP2(308/) (Y) MICE WERE MORE SENSITIVE TO ETHANOL'S STIMULATORY AND SEDATIVE EFFECTS THAN WILD-TYPE (WT) MICE, DRANK LESS ETHANOL IN A LIMITED ACCESS 2 BOTTLE CHOICE PARADIGM AND DID NOT SHOW INCREASED DRINKING AFTER INDUCTION OF DEPENDENCE WITH EXPOSURE TO CIE VAPORS. ALCOHOL METABOLISM DID NOT DIFFER IN MECP2(308/) (Y) AND WT MICE. ADDITIONALLY, MECP2(308/) (Y) MICE DID NOT DIFFER FROM WT MICE IN ETHANOL PREFERENCE IN A 24-HOUR PARADIGM NOR IN THEIR INTAKE OF GRADED SOLUTIONS OF SACCHARIN OR QUININE, SUGGESTING THAT THE MECP2(308/) (Y) MUTATION DID NOT ALTER TASTE FUNCTION. LASTLY, USING THE GENE SET ENRICHMENT ANALYSIS ALGORITHM, WE FOUND A SIGNIFICANT OVERLAP IN THE GENES REGULATED BY ALCOHOL AND BY MECP2. TOGETHER, THESE RESULTS SUGGEST THAT MECP2 CONTRIBUTES TO THE REGULATION OF ETHANOL SENSITIVITY AND DRINKING.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
12   219  31 ACUTE IMMOBILIZATION STRESS FOLLOWING CONTEXTUAL FEAR CONDITIONING REDUCES FEAR MEMORY: TIMING IS ESSENTIAL. BACKGROUND: HISTONE ACETYLATION IS REGULATED IN RESPONSE TO STRESS AND PLAYS AN IMPORTANT ROLE IN LEARNING AND MEMORY. CHRONIC STRESS IS KNOWN TO DETERIORATE COGNITION, WHEREAS ACUTE STRESS FACILITATES MEMORY FORMATION. HOWEVER, WHETHER ACUTE STRESS FACILITATES MEMORY FORMATION WHEN IT IS APPLIED AFTER FEAR STIMULATION IS NOT YET KNOWN. THEREFORE, THIS STUDY AIMED TO INVESTIGATE THE EFFECT OF ACUTE STRESS APPLIED AFTER FEAR TRAINING ON MEMORY FORMATION, MRNA EXPRESSION OF BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF), EPIGENETIC REGULATION OF BDNF EXPRESSION, AND CORTICOSTERONE LEVEL IN MICE IN VIVO. METHODS: MICE WERE SUBJECTED TO ACUTE IMMOBILIZATION STRESS FOR 30 MIN AT 60 OR 90 MIN AFTER CONTEXTUAL FEAR CONDITIONING TRAINING, AND ACETYLATION OF HISTONE 3 AT LYSINE 14 (H3K14) AND LEVEL OF CORTICOSTERONE WERE MEASURED USING WESTERN BLOT ANALYSIS AND ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA), RESPECTIVELY. A FREEZING BEHAVIOR TEST WAS PERFORMED 24 H AFTER TRAINING, AND MRNA EXPRESSION OF BDNF WAS MEASURED USING REAL-TIME POLYMERASE CHAIN REACTIONS. DIFFERENT GROUPS OF MICE WERE USED FOR EACH TEST. RESULTS: FREEZING BEHAVIOR SIGNIFICANTLY DECREASED WITH THE DOWN-REGULATION OF BDNF MRNA EXPRESSION CAUSED BY ACUTE IMMOBILIZATION STRESS AT 60 MIN AFTER FEAR CONDITIONING TRAINING OWING TO THE REDUCTION OF H3K14 ACETYLATION. HOWEVER, BDNF MRNA EXPRESSION AND H3K14 ACETYLATION WERE NOT REDUCED IN ANIMALS SUBJECTED TO IMMOBILIZATION STRESS AT 90 MIN AFTER THE TRAINING. FURTHER, THE CORTICOSTERONE LEVEL WAS SIGNIFICANTLY HIGH IN MICE SUBJECTED TO IMMOBILIZATION STRESS AT 60 MIN AFTER THE TRAINING. CONCLUSION: ACUTE IMMOBILIZATION STRESS FOR 30 MIN AT 60 MIN AFTER FEAR CONDITIONING TRAINING IMPAIRED MEMORY FORMATION AND REDUCED BDNF MRNA EXPRESSION AND H3K14 ACETYLATION IN THE HIPPOCAMPUS OF MICE OWING TO THE HIGH LEVEL OF CORTICOSTERONE.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
13  2243  24 EPIGENETIC MODULATION OF CHRONIC ANXIETY AND PAIN BY HISTONE DEACETYLATION. PROLONGED EXPOSURE OF THE CENTRAL AMYGDALA (CEA) TO ELEVATED CORTICOSTEROIDS (CORT) FACILITATES LONG-TERM ANXIETY AND PAIN THROUGH ACTIVATION OF GLUCOCORTICOID RECEPTORS (GRS) AND CORTICOTROPIN-RELEASING FACTOR (CRF). HOWEVER, THE MECHANISMS MAINTAINING THESE RESPONSES ARE UNKNOWN. SINCE CHRONIC PHENOTYPES CAN BE SUSTAINED BY EPIGENETIC MECHANISMS, INCLUDING HISTONE MODIFICATIONS SUCH AS DEACETYLATION, WE TESTED THE HYPOTHESIS THAT HISTONE DEACETYLATION CONTRIBUTES TO THE MAINTENANCE OF CHRONIC ANXIETY AND PAIN INDUCED BY PROLONGED EXPOSURE OF THE CEA TO CORT. WE FOUND THAT BILATERAL INFUSIONS OF A HISTONE DEACETYLASE INHIBITOR INTO THE CEA ATTENUATED ANXIETY-LIKE BEHAVIOR AS WELL AS SOMATIC AND VISCERAL HYPERSENSITIVITY RESULTING FROM ELEVATED CORT EXPOSURE. MOREOVER, WE DELINEATED A NOVEL PATHWAY THROUGH WHICH HISTONE DEACETYLATION COULD CONTRIBUTE TO CORT REGULATION OF GR AND SUBSEQUENT CRF EXPRESSION IN THE CEA. SPECIFICALLY, DEACETYLATION OF HISTONE 3 AT LYSINE 9 (H3K9), THROUGH THE COORDINATED ACTION OF THE NAD+-DEPENDENT PROTEIN DEACETYLASE SIRTUIN-6 (SIRT6) AND NUCLEAR FACTOR KAPPA B (NFKAPPAB), SEQUESTERS GR EXPRESSION LEADING TO DISINHIBITION OF CRF. OUR RESULTS INDICATE THAT EPIGENETIC PROGRAMMING IN THE AMYGDALA, SPECIFICALLY HISTONE MODIFICATIONS, IS IMPORTANT IN THE MAINTENANCE OF CHRONIC ANXIETY AND PAIN.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
14  1831  37 EFFECTS OF MATERNAL SEPARATION AND ANTIDEPRESSANT DRUG ON EPIGENETIC REGULATION OF THE BRAIN-DERIVED NEUROTROPHIC FACTOR EXON I PROMOTER IN THE ADULT RAT HIPPOCAMPUS. AIM: EARLY LIFE STRESS CAN INDUCE EPIGENETIC CHANGES THROUGH GENETIC AND ENVIRONMENTAL INTERACTIONS AND IS A RISK FACTOR FOR DEPRESSION. BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) HAS BEEN IMPLICATED IN THE PATHOPHYSIOLOGY OF DEPRESSION AND ANTIDEPRESSANT DRUG ACTION. WE INVESTIGATED EPIGENETIC CHANGES AT THE BDNF EXON I PROMOTER IN THE HIPPOCAMPUS OF ADULT RATS SUBJECTED TO MATERNAL SEPARATION (MS) DURING EARLY LIFE AND TREATED WITH AN ANTIDEPRESSANT DRUG AS ADULTS. METHODS: RAT PUPS WERE SUBJECTED TO MS FROM POSTNATAL DAY 1 TO 21 AND RECEIVED CHRONIC ESCITALOPRAM (ESC) AS ADULTS. WE ASSESSED THE EFFECTS OF MS AND ESC ON BDNF EXON I AND DNA METHYLTRANSFERASES (DNMT) MRNA LEVELS (QUANTITATIVE REVERSE-TRANSCRIPTION POLYMERASE CHAIN REACTION), ACETYLATED HISTONE H3, AND MECP2 BINDING TO THE BDNF PROMOTER I (CHROMATIN IMMUNOPRECIPITATION FOLLOWED BY REAL-TIME POLYMERASE CHAIN REACTION), AND BDNF PROTEIN LEVELS (ENZYME-LINKED IMMUNOSORBENT ASSAY). RESULTS: THE LEVELS OF BDNF PROTEIN, EXON I MRNA, HISTONE H3 ACETYLATION, AND DNMT1 AND DNMT3A MRNA WERE ALTERED IN THE MS GROUP COMPARED WITH THE CONTROL GROUP. SIGNIFICANT DECREASES WERE OBSERVED IN THE BDNF PROTEIN, EXON I MRNA, AND HISTONE H3 ACETYLATION LEVELS AND THERE WERE SIGNIFICANT INCREASES IN DNMT1 AND DNMT3A MRNA LEVELS. THE COMPARISON BETWEEN THE MS + ESC AND MS GROUPS REVEALED SIGNIFICANT INCREASES IN BDNF PROTEIN, EXON I MRNA, AND HISTONE H3 ACETYLATION LEVELS AND SIGNIFICANT DECREASES IN MECP2 AND DNMT1 AND DNMT3A MRNA LEVELS. CONCLUSION: THESE FINDINGS INDICATE THAT MS INDUCED EPIGENETIC CHANGES AT THE BDNF EXON I PROMOTER AND THESE CHANGES WERE PREVENTED BY ANTIDEPRESSANT DRUG TREATMENT DURING ADULTHOOD.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
15  2300  31 EPIGENETIC REGULATION OF BDNF EXPRESSION IN THE PRIMARY SENSORY NEURONS AFTER PERIPHERAL NERVE INJURY: IMPLICATIONS IN THE DEVELOPMENT OF NEUROPATHIC PAIN. BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) IS KNOWN TO BE UP-REGULATED IN THE DORSAL ROOT GANGLION (DRG) AFTER PERIPHERAL NERVE INJURY, AND TO CONTRIBUTE TO NEUROPATHIC PAIN. HERE, WE FOUND THAT THERMAL HYPERALGESIA AND MECHANICAL ALLODYNIA AT DAY 7 POST-INJURY WERE INHIBITED ONLY WHEN ANTI-BDNF ANTIBODY WAS INTRATHECALLY ADMINISTRATED AT DAY 2 POST-INJURY. CONSISTENT WITH BEHAVIORAL RESULTS, WESTERN BLOT ANALYSIS SHOWED THAT THE EXPRESSION LEVELS OF BDNF PROTEIN IN THE SPINAL DORSAL HORN WERE MARKEDLY INDUCED DURING EARLY STAGE POST-INJURY. MOREOVER, THE MAXIMAL INCREASE IN BDNF MRNA EXPRESSION IN THE DRG WAS OBSERVED AT DAY 1 POST-INJURY, AND SIGNIFICANTLY ELEVATED LEVELS WERE SUSTAINED FOR AT LEAST 14 DAYS. FOUR OF FIVE BDNF MRNA TRANSCRIPTS WERE UP-REGULATED AFTER NERVE INJURY, AND THE MOST INDUCIBLE TRANSCRIPT WAS EXON I. USING A CHROMATIN IMMUNOPRECIPITATION (CHIP) ASSAY, WE FOUND THAT NERVE INJURY PROMOTES HISTONE H3 AND H4 ACETYLATION, TRANSCRIPTIONALLY ACTIVE MODIFICATIONS, AT BDNF PROMOTER I AT DAY 1 POST-INJURY, AND THE LEVELS OF HISTONE ACETYLATION REMAIN ELEVATED FOR AT LEAST 7 DAYS. TAKEN TOGETHER, OUR FINDINGS SUGGEST THAT AN INITIAL INCREASE IN BDNF EXON I EXPRESSION CONTROLLED BY EPIGENETIC MECHANISMS MIGHT HAVE A CRUCIAL ROLE IN THE DEVELOPMENT OF NEUROPATHIC PAIN.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
16  3082  40 GENOME-WIDE REDISTRIBUTION OF MECP2 IN DORSAL ROOT GANGLIA AFTER PERIPHERAL NERVE INJURY. BACKGROUND: METHYL-CPG-BINDING PROTEIN 2 (MECP2), A PROTEIN WITH AFFINITY FOR METHYLATED CYTOSINES, IS CRUCIAL FOR NEURONAL DEVELOPMENT AND FUNCTION. MECP2 REGULATES GENE EXPRESSION THROUGH ACTIVATION, REPRESSION AND CHROMATIN REMODELING. MUTATIONS IN MECP2 CAUSE RETT SYNDROME, AND THESE PATIENTS DISPLAY IMPAIRED NOCICEPTION. WE OBSERVED AN INCREASE IN MECP2 EXPRESSION IN MOUSE DORSAL ROOT GANGLIA (DRG) AFTER PERIPHERAL NERVE INJURY. THE FUNCTIONAL IMPLICATION OF INCREASED MECP2 IS LARGELY UNKNOWN. TO IDENTIFY REGIONS OF THE GENOME BOUND BY MECP2 IN THE DRG AND THE CHANGES INDUCED BY NERVE INJURY, A CHROMATIN IMMUNOPRECIPITATION OF MECP2 FOLLOWED BY SEQUENCING (CHIP-SEQ) WAS PERFORMED 4 WEEKS AFTER SPARED NERVE INJURY (SNI). RESULTS: WHILE THE NUMBER OF BINDING SITES ACROSS THE GENOME REMAINED SIMILAR IN THE SNI MODEL AND SHAM CONTROL, SNI INDUCED THE REDISTRIBUTION OF MECP2 TO TRANSCRIPTIONALLY RELEVANT REGIONS. TO DETERMINE HOW DIFFERENTIAL BINDING OF MECP2 CAN AFFECT GENE EXPRESSION IN THE DRG, WE INVESTIGATED MMU-MIR-126, A MICRORNA LOCUS THAT HAD ENRICHED MECP2 BINDING IN THE SNI MODEL. ENRICHED MECP2 BINDING TO MIR-126 LOCUS AFTER NERVE INJURY REPRESSED MIR-126 EXPRESSION, AND THIS WAS NOT MEDIATED BY ALTERATIONS IN METHYLATION PATTERN AT THE MIR-126 LOCUS. DOWNREGULATION OF MIR-126 RESULTED IN THE UPREGULATION OF ITS TWO TARGET GENES DNMT1 AND VEGFA IN NEURO 2A CELLS AND IN SNI MODEL COMPARED TO CONTROL. THESE TARGET GENES WERE SIGNIFICANTLY DOWNREGULATED IN MECP2-NULL MICE COMPARED TO WILD-TYPE LITTERMATES, INDICATING A REGULATORY ROLE FOR MECP2 IN ACTIVATING DNMT1 AND VEGFA EXPRESSION. INTRATHECAL DELIVERY OF MIR-126 WAS NOT SUFFICIENT TO REVERSE NERVE INJURY-INDUCED MECHANICAL AND THERMAL HYPERSENSITIVITY, BUT DECREASED DNMT1 AND VEGFA EXPRESSION IN THE DRG. CONCLUSIONS: OUR STUDY SHOWS A REGULATORY ROLE FOR MECP2 IN THAT CHANGES IN GLOBAL REDISTRIBUTION CAN RESULT IN DIRECT AND INDIRECT MODULATION OF GENE EXPRESSION IN THE DRG. ALTERATIONS IN GENOME-WIDE BINDING OF MECP2 THEREFORE PROVIDE A MOLECULAR BASIS FOR A BETTER UNDERSTANDING OF EPIGENETIC REGULATION-INDUCED MOLECULAR CHANGES UNDERLYING NERVE INJURY.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                       
17  5712  39 SIRT1 MEDIATES DEPRESSION-LIKE BEHAVIORS IN THE NUCLEUS ACCUMBENS. DEPRESSION IS A RECURRING AND LIFE-THREATENING ILLNESS THAT AFFECTS UP TO 120 MILLION PEOPLE WORLDWIDE. IN THE PRESENT STUDY, WE SHOW THAT CHRONIC SOCIAL DEFEAT STRESS, AN ETHOLOGICALLY VALIDATED MODEL OF DEPRESSION IN MICE, INCREASES SIRT1 LEVELS IN THE NUCLEUS ACCUMBENS (NAC), A KEY BRAIN REWARD REGION. INCREASES IN SIRT1, A WELL CHARACTERIZED CLASS III HISTONE DEACETYLASE, AFTER CHRONIC SOCIAL DEFEAT SUGGEST A ROLE FOR THIS ENZYME IN MEDIATING DEPRESSION-LIKE BEHAVIORS. WHEN RESVERATROL, A PHARMACOLOGICAL ACTIVATOR OF SIRT1, WAS DIRECTLY INFUSED BILATERALLY INTO THE NAC, WE OBSERVED AN INCREASE IN DEPRESSION- AND ANXIETY-LIKE BEHAVIORS. CONVERSELY, INTRA-NAC INFUSIONS OF EX-527, A SIRT1 ANTAGONIST, REDUCED THESE BEHAVIORS; EX-527 ALSO REDUCED ACUTE STRESS RESPONSES IN STRESS-NAIVE MICE. NEXT, WE INCREASED SIRT1 LEVELS DIRECTLY IN NAC BY USE OF VIRAL-MEDIATED GENE TRANSFER AND OBSERVED AN INCREASE IN DEPRESSIVE- AND ANXIETY-LIKE BEHAVIORS WHEN MICE WERE ASSESSED IN THE OPEN-FIELD, ELEVATED-PLUS-MAZE, AND FORCED SWIM TESTS. USING A CRE-INDUCIBLE VIRAL VECTOR SYSTEM TO OVEREXPRESS SIRT1 SELECTIVELY IN DOPAMINE D1 OR D2 SUBPOPULATIONS OF MEDIUM SPINY NEURONS (MSNS) IN THE NAC, WE FOUND THAT SIRT1 PROMOTES DEPRESSIVE-LIKE BEHAVIORS ONLY WHEN OVEREXPRESSED IN D1 MSNS, WITH NO EFFECT SEEN IN D2 MSNS. CONVERSELY, SELECTIVE ABLATION OF SIRT1 IN THE NAC USING VIRAL-CRE IN FLOXED SIRT1 MICE RESULTED IN DECREASED DEPRESSION- AND ANXIETY-LIKE BEHAVIORS. TOGETHER, THESE RESULTS DEMONSTRATE THAT SIRT1 PLAYS AN ESSENTIAL ROLE IN THE NAC IN REGULATING MOOD-RELATED BEHAVIORAL ABNORMALITIES AND IDENTIFIES A NOVEL SIGNALING PATHWAY FOR THE DEVELOPMENT OF INNOVATIVE ANTIDEPRESSANTS TO TREAT MAJOR DEPRESSIVE DISORDERS. SIGNIFICANCE STATEMENT: IN THIS STUDY, WE DEMONSTRATE A PIVOTAL ROLE FOR SIRT1 IN ANXIETY- AND DEPRESSION-LIKE BEHAVIORS IN THE NUCLEUS ACCUMBENS (NAC), A KEY BRAIN REWARD REGION. WE SHOW THAT STRESS STABLY INDUCES SIRT1 EXPRESSION IN THIS BRAIN REGION AND THAT ALTERING SIRT1 ACTIVITY USING A PHARMACOLOGICAL OR GENETIC APPROACH REGULATES ANXIETY- AND DEPRESSION-LIKE BEHAVIORS. THESE RESULTS SUGGEST THAT SIRT1 PLAYS AN ESSENTIAL ROLE IN REGULATING MOOD-RELATED BEHAVIORS AND INTRODUCES A NOVEL SIGNALING PATHWAY FOR THE DEVELOPMENT OF INNOVATIVE ANTIDEPRESSANTS TO TREAT DEPRESSION AND OTHER STRESS-RELATED DISORDERS. A RECENT GROUNDBREAKING PUBLICATION BY THE CONVERGE CONSORTIUM (2015) IDENTIFIED A REPRODUCIBLE ASSOCIATION OF THE SIRT1 LOCUS WITH MAJOR DEPRESSION IN HUMANS. THEREFORE, OUR RESULTS ARE TIMELY AND HAVE SIGNIFICANT TRANSLATIONAL RELEVANCE.	2016	
           
18  4303  35 MICRORNA-223 INHIBITS TISSUE FACTOR EXPRESSION IN VASCULAR ENDOTHELIAL CELLS. OBJECTIVE: ATHEROSCLEROSIS IS A CHRONIC INFLAMMATORY PROCESS, IN WHICH VASCULAR ENDOTHELIAL CELLS (ECS) BECOME DYSFUNCTIONAL OWING TO THE EFFECTS OF CHEMICAL SUBSTANCES, SUCH AS INFLAMMATORY FACTOR AND GROWTH FACTORS. TISSUE FACTOR (TF) EXPRESSION IS INDUCED BY THE ABOVE CHEMICAL SUBSTANCES IN ACTIVATED ECS. TF INITIATES THROMBOSIS ON DISRUPTED ATHEROSCLEROTIC PLAQUES WHICH PLAYS AN ESSENTIAL ROLE DURING THE ONSET OF ACUTE CORONARY SYNDROMES (ACS). INCREASING EVIDENCES SUGGEST THE IMPORTANT ROLE OF MICRORNAS AS EPIGENETIC REGULATORS OF ATHEROSCLEROTIC DISEASE. THE AIM OF OUR STUDY IS TO IDENTIFY IF MICRORNA-223 (MIR-223) TARGETS TF IN ECS. METHODS AND RESULTS: BIOINFORMATIC ANALYSIS SHOWED THAT TF IS A TARGET CANDIDATE OF MIR-223. WESTERN BLOTTING ANALYSIS REVEALED THAT TUMOR NECROSIS FACTOR ALPHA (TNF-ALPHA) INCREASED TF EXPRESSION IN AORTA OF C57BL/6J MICE AND CULTURED ECS (EA.HY926 CELLS AND HUVEC) AFTER 4 H TREATMENT. IN TNF-ALPHA TREATED ECS, TF MRNA WAS ALSO INCREASED MEASURED BY REAL-TIME PCR. REAL-TIME PCR RESULTS SHOWED THAT MIR-223 LEVELS WERE DOWNREGULATED IN TNF-ALPHA-TREATED AORTA OF C57BL/6J MICE AND CULTURED ECS. TRANSFECTION OF ECS WITH MIR-223 MIMIC OR MIR-223 INHIBITOR MODIFIED TF EXPRESSION BOTH IN MRNA AND PROTEIN LEVELS. LUCIFERASE ASSAYS CONFIRMED THAT MIR-223 SUPPRESSED TF EXPRESSION BY BINDING TO THE SEQUENCE OF TF 3'-UNTRANSLATED REGIONS (3'UTR). TF PROCOAGULANT ACTIVITY WAS INHIBITED BY OVEREXPRESSING MIR-223 WITH OR WITHOUT TNF-ALPHA STIMULATION. CONCLUSIONS: MIR-223-MEDIATED SUPPRESSION OF TF EXPRESSION PROVIDES A NOVEL MOLECULAR MECHANISM FOR THE REGULATION OF COAGULATION CASCADE, AND SUGGESTS A CLUE AGAINST THROMBOGENESIS DURING THE PROCESS OF ATHEROSCLEROTIC PLAQUE RUPTURE.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
19   910  42 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
20  3608  38 IN SEARCH FOR GENES RELATED TO ATHEROSCLEROSIS AND DYSLIPIDEMIA USING ANIMAL MODELS. ATHEROSCLEROSIS IS A MULTIFACTORIAL CHRONIC DISEASE THAT AFFECTS LARGE ARTERIES AND MAY LEAD TO FATAL CONSEQUENCES. ACCORDING TO CURRENT UNDERSTANDING, INFLAMMATION AND LIPID ACCUMULATION ARE THE TWO KEY MECHANISMS OF ATHEROSCLEROSIS DEVELOPMENT. ANIMAL MODELS BASED ON GENETICALLY MODIFIED MICE HAVE BEEN DEVELOPED TO INVESTIGATE THESE ASPECTS. ONE SUCH MODEL IS LOW-DENSITY LIPOPROTEIN (LDL) RECEPTOR KNOCKOUT (KO) MICE (LDLR(-/-)), WHICH ARE CHARACTERIZED BY A MODERATE INCREASE OF PLASMA LDL CHOLESTEROL LEVELS. ANOTHER WIDELY USED GENETICALLY MODIFIED MOUSE STRAIN IS APOLIPOPROTEIN-E KO MICE (APOE(-/-)) THAT LACKS THE PRIMARY LIPOPROTEIN REQUIRED FOR THE UPTAKE OF LIPOPROTEINS THROUGH THE HEPATIC RECEPTORS, LEADING TO EVEN GREATER PLASMA CHOLESTEROL INCREASE THAN IN LDLR(-/-) MICE. THESE AND OTHER ANIMAL MODELS ALLOWED FOR CONDUCTING GENETIC STUDIES, SUCH AS GENOME-WIDE ASSOCIATION STUDIES, MICROARRAYS, AND GENOTYPING METHODS, WHICH HELPED IDENTIFYING MORE THAN 100 MUTATIONS THAT CONTRIBUTE TO ATHEROSCLEROSIS DEVELOPMENT. HOWEVER, TRANSLATION OF THE RESULTS OBTAINED IN ANIMAL MODELS FOR HUMAN SITUATIONS WAS SLOW AND CHALLENGING. AT THE SAME TIME, GENETIC STUDIES CONDUCTED IN HUMANS WERE LIMITED BY LOW SAMPLE SIZES AND HIGH HETEROGENEITY IN PREDICTIVE SUBCLINICAL PHENOTYPES. IN THIS REVIEW, WE SUMMARIZE THE CURRENT KNOWLEDGE ON THE USE OF KO MICE FOR IDENTIFICATION OF GENES IMPLICATED IN ATHEROSCLEROSIS AND PROVIDE A LIST OF GENES INVOLVED IN ATHEROSCLEROSIS-ASSOCIATED INFLAMMATORY PATHWAYS AND THEIR BRIEF CHARACTERISTICS. MOREOVER, WE DISCUSS THE APPROACHES FOR CANDIDATE GENE SEARCH IN ANIMALS AND HUMANS AND DISCUSS THE PROGRESS MADE IN THE FIELD OF EPIGENETIC STUDIES THAT APPEAR TO BE PROMISING FOR IDENTIFICATION OF NOVEL BIOMARKERS AND THERAPEUTIC TARGETS.	2020