1  1105 148 COMBINED INHIBITION OF HISTONE DEACETYLASES AND BET FAMILY PROTEINS AS EPIGENETIC THERAPY FOR NERVE INJURY-INDUCED NEUROPATHIC PAIN. CURRENT TREATMENTS FOR NEUROPATHIC PAIN HAVE OFTEN MODERATE EFFICACY AND PRESENT UNWANTED EFFECTS SHOWING THE NEED TO DEVELOP EFFECTIVE THERAPIES. ACCUMULATING EVIDENCE SUGGESTS THAT HISTONE ACETYLATION PLAYS ESSENTIAL ROLES IN CHRONIC PAIN AND THE ANALGESIC ACTIVITY OF HISTONE DEACETYLASES (HDACS) INHIBITORS IS DOCUMENTED. BROMODOMAIN AND EXTRA-TERMINAL DOMAIN (BET) PROTEINS ARE EPIGENETIC READERS THAT INTERACT WITH ACETYLATED LYSINE RESIDUES ON HISTONES, BUT LITTLE IS KNOWN ABOUT THEIR IMPLICATION IN NEUROPATHIC PAIN. THUS, THE CURRENT STUDY WAS AIMED TO INVESTIGATE THE EFFECT OF THE COMBINATION OF HDAC AND BET INHIBITORS IN THE SPARED NERVE INJURY (SNI) MODEL IN MICE. INTRANASAL ADMINISTRATION OF I-BET762 (BET INHIBITOR) OR SAHA (HDAC INHIBITOR) ATTENUATED THERMAL AND MECHANICAL HYPERSENSITIVITY AND THIS ANTIALLODYNIC ACTIVITY WAS IMPROVED BY CO-ADMINISTRATION OF BOTH DRUGS. SPINAL CORD SECTIONS OF SNI MICE SHOWED AN INCREASED EXPRESSION OF HDAC1 AND BRD4 PROTEINS AND COMBINATION PRODUCED A STRONGER REDUCTION COMPARED TO EACH EPIGENETIC AGENT ALONE. SAHA AND I-BET762, ADMINISTERED ALONE OR IN COMBINATION, COUNTERACTED THE SNI-INDUCED MICROGLIA ACTIVATION BY INHIBITING THE EXPRESSION OF IBA1, CD11B, INDUCIBLE NITRIC OXIDE SYNTHASE (INOS), THE ACTIVATION OF NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) AND SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION-1 (STAT1) WITH COMPARABLE EFFICACY. CONVERSELY, THE EPIGENETIC INHIBITORS SHOWED A MODEST EFFECT ON SPINAL PROINFLAMMATORY CYTOKINES CONTENT THAT WAS SIGNIFICANTLY POTENTIATED BY THEIR COMBINATION. PRESENT RESULTS INDICATE A KEY ROLE OF ACETYLATED HISTONES AND THEIR RECRUITMENT BY BET PROTEINS ON MICROGLIA-MEDIATED SPINAL NEUROINFLAMMATION. TARGETING NEUROPATHIC PAIN WITH THE COMBINATION OF HDAC AND BET INHIBITORS MAY REPRESENT A PROMISING NEW THERAPEUTIC OPTION.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
2  3656  37 INDUCIBLE PRMT1 ABLATION IN ADULT VASCULAR SMOOTH MUSCLE LEADS TO CONTRACTILE DYSFUNCTION AND AORTIC DISSECTION. VASCULAR SMOOTH MUSCLE CELLS (VSMCS) HAVE REMARKABLE PLASTICITY IN RESPONSE TO DIVERSE ENVIRONMENTAL CUES. ALTHOUGH THESE CELLS ARE VERSATILE, CHRONIC STRESS CAN TRIGGER VSMC DYSFUNCTION, WHICH ULTIMATELY LEADS TO VASCULAR DISEASES SUCH AS AORTIC ANEURYSM AND ATHEROSCLEROSIS. PROTEIN ARGININE METHYLTRANSFERASE 1 (PRMT1) IS A MAJOR ENZYME CATALYZING ASYMMETRIC ARGININE DIMETHYLATION OF PROTEINS THAT ARE SOURCES OF ASYMMETRIC DIMETHYLARGININE (ADMA), AN ENDOGENOUS INHIBITOR OF NITRIC OXIDE SYNTHASE. ALTHOUGH A POTENTIAL ROLE OF PRMT1 IN VASCULAR PATHOGENESIS HAS BEEN PROPOSED, ITS ROLE IN VASCULAR FUNCTION HAS YET TO BE CLARIFIED. HERE, WE INVESTIGATED THE ROLE AND UNDERLYING MECHANISM OF PRMT1 IN VASCULAR SMOOTH MUSCLE CONTRACTILITY AND FUNCTION. THE EXPRESSION OF PRMT1 AND CONTRACTILE-RELATED GENES WAS SIGNIFICANTLY DECREASED IN THE AORTAS OF ELDERLY HUMANS AND PATIENTS WITH AORTIC ANEURYSMS. MICE WITH VSMC-SPECIFIC PRMT1 ABLATION (SMKO) EXHIBITED PARTIAL LETHALITY, LOW BLOOD PRESSURE AND AORTIC DILATION. THE PRMT1-ABLATED AORTAS SHOWED AORTIC DISSECTION WITH ELASTIC FIBER DEGENERATION AND CELL DEATH. EX VIVO AND IN VITRO ANALYSES INDICATED THAT PRMT1 ABLATION SIGNIFICANTLY DECREASED THE CONTRACTILITY OF THE AORTA AND TRACTION FORCES OF VSMCS. PRMT1 ABLATION DOWNREGULATED THE EXPRESSION OF CONTRACTILE GENES SUCH AS MYOCARDIN WHILE UPREGULATING THE EXPRESSION OF SYNTHETIC GENES, THUS CAUSING THE CONTRACTILE TO SYNTHETIC PHENOTYPIC SWITCH OF VSMCS. IN ADDITION, MECHANISTIC STUDIES DEMONSTRATED THAT PRMT1 DIRECTLY REGULATES MYOCARDIN GENE ACTIVATION BY MODULATING EPIGENETIC HISTONE MODIFICATIONS IN THE MYOCARDIN PROMOTER REGION. THUS, OUR STUDY DEMONSTRATES THAT VSMC PRMT1 IS ESSENTIAL FOR VASCULAR HOMEOSTASIS AND THAT ITS ABLATION CAUSES AORTIC DILATION/DISSECTION THROUGH IMPAIRED MYOCARDIN EXPRESSION.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
3   351  30 ALTERED ENDOTHELIAL DYSFUNCTION-RELATED MIRS IN PLASMA FROM ME/CFS PATIENTS. MYALGIC ENCEPHALOMYELITIS/CHRONIC FATIGUE SYNDROME (ME/CFS) IS A COMPLEX DISEASE CHARACTERIZED BY UNEXPLAINED DEBILITATING FATIGUE. ALTHOUGH THE ETIOLOGY IS UNKNOWN, EVIDENCE SUPPORTS IMMUNOLOGICAL ABNORMALITIES, SUCH AS PERSISTENT INFLAMMATION AND IMMUNE-CELL ACTIVATION, IN A SUBSET OF PATIENTS. SINCE THE INTERPLAY BETWEEN INFLAMMATION AND VASCULAR ALTERATIONS IS WELL-ESTABLISHED IN OTHER DISEASES, ENDOTHELIAL DYSFUNCTION HAS EMERGED AS ANOTHER PLAYER IN ME/CFS PATHOGENESIS. ENDOTHELIAL NITRIC OXIDE SYNTHASE (ENOS) GENERATES NITRIC OXIDE (NO) THAT MAINTAINS ENDOTHELIAL HOMEOSTASIS. ENOS IS ACTIVATED BY SILENT INFORMATION REGULATOR 1 (SIRT1), AN ANTI-INFLAMMATORY PROTEIN. DESPITE ITS RELEVANCE, NO STUDY HAS ADDRESSED THE SIRT1/ENOS AXIS IN ME/CFS. THE INTEREST IN CIRCULATING MICRORNAS (MIRS) AS POTENTIAL BIOMARKERS IN ME/CFS HAS INCREASED IN RECENT YEARS. ACCORDINGLY, WE ANALYZE A SET OF MIRS REPORTED TO MODULATE THE SIRT1/ENOS AXIS USING PLASMA FROM ME/CFS PATIENTS. OUR RESULTS SHOW THAT MIR-21, MIR-34A, MIR-92A, MIR-126, AND MIR-200C ARE JOINTLY INCREASED IN ME/CFS PATIENTS COMPARED TO HEALTHY CONTROLS. A SIMILAR FINDING WAS OBTAINED WHEN ANALYZING PUBLIC MIR DATA ON PERIPHERAL BLOOD MONONUCLEAR CELLS. BIOINFORMATICS ANALYSIS SHOWS THAT ENDOTHELIAL FUNCTION-RELATED SIGNALING PATHWAYS ARE ASSOCIATED WITH THESE MIRS, INCLUDING OXIDATIVE STRESS AND OXYGEN REGULATION. INTERESTINGLY, HISTONE DEACETYLASE 1, A PROTEIN RESPONSIBLE FOR EPIGENETIC REGULATIONS, REPRESENTED THE MOST RELEVANT NODE WITHIN THE NETWORK. IN CONCLUSION, OUR STUDY PROVIDES A BASIS TO FIND ENDOTHELIAL DYSFUNCTION-RELATED BIOMARKERS AND EXPLORE NOVEL TARGETS IN ME/CFS.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
4  5479  31 RESVERATROL ATTENUATES CIGARETTE SMOKE EXTRACT INDUCED CELLULAR SENESCENCE IN HUMAN AIRWAY EPITHELIAL CELLS BY REGULATING THE MIR-34A/SIRT1/NF-KAPPAB PATHWAY. CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) IS CHARACTERIZED BY ACCELERATED LUNG AGING. SMOKING IS THE CRITICAL RISK FACTOR FOR COPD. CELLULAR SENESCENCE OF AIRWAY EPITHELIAL CELLS IS THE CYTOLOGICAL BASIS OF ACCELERATED LUNG AGING IN COPD, AND THE REGULATION OF MICRORNAS (MIRNAS) IS THE CENTRAL EPIGENETIC MECHANISM OF CELLULAR SENESCENCE. RESVERATROL (RES) IS A POLYPHENOL WITH ANTI-AGING PROPERTIES. THIS STUDY INVESTIGATED WHETHER RES ATTENUATES CIGARETTE SMOKE EXTRACT (CSE)-INDUCED CELLULAR SENESCENCE IN HUMAN AIRWAY EPITHELIAL CELLS (BEAS-2B) THROUGH THE MIR-34A/SIRT1/NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) PATHWAY. BEAS-2B CELLS WERE TREATED WITH RES, CSE AND TRANSFECTED WITH MIR-34A-5P MIMICS. CELLULAR SENESCENCE WAS EVALUATED BY SENESCENCE -RELATED BETA-GALACTOSIDASE (SA-BETA-GAL) STAINING AND EXPRESSION OF SENESCENCE-RELATED GENES (P16, P21, AND P53). THE EXPRESSIONS OF MIR-34A-5P, SIRT1, AND NF-KAPPAB P65 WERE EXAMINED USING QUANTITATIVE REAL TIME POLYMERASE CHAIN REACTION AND WESTERN BLOTTING. THE SENESCENCE-ASSOCIATED SECRETORY PHENOTYPE (SASP) CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) WERE ASSESSED BY ENZYME-LINKED IMMUNOSORBENT ASSAY. THE BINDING BETWEEN MIR-34A-5P AND SIRT1 WAS CONFIRMED BY DUAL-LUCIFERASE REPORTER ASSAY. THE RESULTS SHOWED THAT CSE DOSE-DEPENDENTLY DECREASED CELL VIABILITY AND ELEVATED CELLULAR SENESCENCE, CHARACTERIZED BY INCREASED SA-BETA-GAL STAINING AND SENESCENCE-RELATED GENE EXPRESSIONS (P16, P21, AND P53). FURTHER, CSE DOSE-DEPENDENTLY INCREASED THE EXPRESSION OF MIR-34A-5P AND SASP CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) IN BEAS-2B CELLS. PRETREATMENT WITH RES INHIBITED CSE-INDUCED CELLULAR SENESCENCE AND SECRETION OF SASP CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) IN A DOSE-DEPENDENT MANNER. MOREOVER, RES REVERSED THE CSE-INDUCED DOWN-REGULATION OF SIRT1 AND UP-REGULATION OF MIR-34A-5P AND NF-KAPPAB P65. SIRT1 IS A TARGET OF MIR-34A-5P. OVEREXPRESSION OF MIR-34A-5P VIA TRANSFECTION WITH MIR-34A-5P MIMIC IN BEAS-2B CELLS ATTENUATED THE INHIBITORY EFFECT OF RES ON CELLULAR SENESCENCE, ACCOMPANIED BY REVERSING THE EXPRESSION OF SIRT1 AND NF-KAPPAB P65. IN CONCLUSION, RES ATTENUATED CSE-INDUCED CELLULAR SENESCENCE IN BEAS-2B CELLS BY REGULATING THE MIR-34A/SIRT1/NF-KAPPAB PATHWAY, WHICH MAY PROVIDE A NEW APPROACH FOR COPD TREATMENT.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                       
5  4582  43 N-TERMINAL BET BROMODOMAIN INHIBITORS DISRUPT A BRD4-P65 INTERACTION AND REDUCE INDUCIBLE NITRIC OXIDE SYNTHASE TRANSCRIPTION IN PANCREATIC BETA-CELLS. CHRONIC INFLAMMATION OF PANCREATIC ISLETS IS A KEY DRIVER OF BETA-CELL DAMAGE THAT CAN LEAD TO AUTOREACTIVITY AND THE EVENTUAL ONSET OF AUTOIMMUNE DIABETES (T1D). IN THE ISLET, ELEVATED LEVELS OF PROINFLAMMATORY CYTOKINES INDUCE THE TRANSCRIPTION OF THE INDUCIBLE NITRIC OXIDE SYNTHASE (INOS) GENE, NOS2, ULTIMATELY RESULTING IN INCREASED NITRIC OXIDE (NO). EXCESSIVE OR PROLONGED EXPOSURE TO NO CAUSES BETA-CELL DYSFUNCTION AND FAILURE ASSOCIATED WITH DEFECTS IN MITOCHONDRIAL RESPIRATION. RECENT STUDIES SHOWED THAT INHIBITION OF THE BROMODOMAIN AND EXTRATERMINAL DOMAIN (BET) FAMILY OF PROTEINS, A DRUGGABLE CLASS OF EPIGENETIC READER PROTEINS, PREVENTS THE ONSET AND PROGRESSION OF T1D IN THE NON-OBESE DIABETIC MOUSE MODEL. WE HYPOTHESIZED THAT BET PROTEINS CO-ACTIVATE TRANSCRIPTION OF CYTOKINE-INDUCED INFLAMMATORY GENE TARGETS IN BETA-CELLS AND THAT SELECTIVE, CHEMOTHERAPEUTIC INHIBITION OF BET BROMODOMAINS COULD REDUCE SUCH TRANSCRIPTION. HERE, WE INVESTIGATED THE ABILITY OF BET BROMODOMAIN SMALL MOLECULE INHIBITORS TO REDUCE THE BETA-CELL RESPONSE TO THE PROINFLAMMATORY CYTOKINE INTERLEUKIN 1 BETA (IL-1BETA). BET BROMODOMAIN INHIBITION ATTENUATED IL-1BETA-INDUCED TRANSCRIPTION OF THE INFLAMMATORY MEDIATOR NOS2 AND CONSEQUENT INOS PROTEIN AND NO PRODUCTION. REDUCED NOS2 TRANSCRIPTION IS CONSISTENT WITH INHIBITION OF NF-KAPPAB FACILITATED BY DISRUPTING THE INTERACTION OF A SINGLE BET FAMILY MEMBER, BRD4, WITH THE NF-KAPPAB SUBUNIT, P65. USING RECENTLY REPORTED SELECTIVE INHIBITORS OF THE FIRST AND SECOND BET BROMODOMAINS, INHIBITION OF ONLY THE FIRST BROMODOMAIN WAS NECESSARY TO REDUCE THE INTERACTION OF BRD4 WITH P65 IN BETA-CELLS. MOREOVER, INHIBITION OF THE FIRST BROMODOMAIN WAS SUFFICIENT TO MITIGATE IL-1BETA-DRIVEN DECREASES IN MITOCHONDRIAL OXYGEN CONSUMPTION RATES AND BETA-CELL VIABILITY. BY IDENTIFYING A ROLE FOR THE INTERACTION BETWEEN BRD4 AND P65 IN CONTROLLING THE RESPONSE OF BETA-CELLS TO PROINFLAMMATORY CYTOKINES, WE PROVIDE MECHANISTIC INFORMATION ON HOW BET BROMODOMAIN INHIBITION CAN DECREASE INFLAMMATION. THESE STUDIES ALSO SUPPORT THE POTENTIAL THERAPEUTIC APPLICATION OF MORE SELECTIVE BET BROMODOMAIN INHIBITORS IN ATTENUATING BETA-CELL INFLAMMATION.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
6  1126  38 COMPLEX REGULATION OF THE REGULATOR OF SYNAPTIC PLASTICITY HISTONE DEACETYLASE 2 IN THE RODENT DORSAL HORN AFTER PERIPHERAL INJURY. HISTONE DEACETYLASES (HDACS), HDAC2 IN PARTICULAR, HAVE BEEN SHOWN TO REGULATE VARIOUS FORMS OF LEARNING AND MEMORY. SINCE COGNITIVE PROCESSES SHARE MECHANISMS WITH SPINAL NOCICEPTIVE SIGNALLING, WE DECIDED TO INVESTIGATE THE HDAC2 EXPRESSION IN THE DORSAL HORN AFTER PERIPHERAL INJURY. USING IMMUNOHISTOCHEMISTRY, WE FOUND THAT SPINAL HDAC2 WAS MAINLY SEEN IN NEURONS AND ASTROCYTES, WITH NEURONAL EXPRESSION IN NAIVE TISSUE 2.6 TIMES GREATER THAN THAT IN ASTROCYTES. CYSTEINE (S)-NITROSYLATION OF HDAC2 RELEASES HDAC2 GENE SILENCING AND IS CONTROLLED BY NITRIC OXIDE (NO). A DURATION OF 48 H AFTER INTRAPLANTAR INJECTION OF COMPLETE FREUND'S ADJUVANT, THERE WAS AN IPSILATERAL INCREASE IN THE MOST IMPORTANT NO-PRODUCING ENZYME IN PAIN STATES, NITRIC OXIDE SYNTHASE (NNOS), ACCOMPANIED BY AN INCREASE IN HDAC2 S-NITROSYLATION. MOREOVER, A SUBSET OF NNOS-POSITIVE NEURONS EXPRESSED CFOS, A KNOWN TARGET OF HDAC2, SUGGESTING THAT DEREPRESSION OF CFOS EXPRESSION FOLLOWING HDAC2 S-NITROSYLATION MIGHT OCCUR AFTER NOXIOUS STIMULATION. WE SAW NO CHANGE IN GLOBAL HDAC2 EXPRESSION IN BOTH SHORT- AND LONG-TERM PAIN STATES. HOWEVER, HDAC2 WAS INCREASED IN ASTROCYTES 7 DAYS AFTER NEUROPATHIC INJURY SUGGESTING THAT HDAC2 MIGHT INHIBIT ASTROCYTIC GENE EXPRESSION IN NEUROPATHIC PAIN STATES. ALL TOGETHER, OUR RESULTS INDICATE THAT THE EPIGENETIC REGULATION OF TRANSCRIPTIONAL PROGRAMMES IN THE DORSAL HORN AFTER INJURY IS CELL SPECIFIC. MOREOVER, THE PROMINENT ROLE OF NO IN PERSISTENT PAIN STATES SUGGESTS THAT HDAC2 S-NITROSYLATION COULD PLAY A CRUCIAL ROLE IN THE REGULATION OF GENE EXPRESSION LEADING TO HYPERSENSITIVITY. OUR MANUSCRIPT DESCRIBES FOR THE FIRST TIME THE REGULATION OF THE MEMORY REGULATOR HISTONE DEACETYLASE 2 (HDAC2) IN THE SUPERFICIAL DORSAL HORN OF ADULT RATS FOLLOWING PERIPHERAL INJURY. OUR CELL-SPECIFIC APPROACH HAS REVEALED A COMPLEX PATTERN OF EXPRESSION OF SPINAL HDAC2 THAT DEPENDS ON THE INJURY AND THE CELL TYPE, SUGGESTING A SOPHISTICATED REGULATION OF GENE EXPRESSION BY HDAC2.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
7  3624  35 IN VIVO FUNCTION OF FLOW-RESPONSIVE CIS-DNA ELEMENTS OF ENOS GENE: A ROLE FOR CHROMATIN-BASED MECHANISMS. BACKGROUND: ENOS (ENDOTHELIAL NITRIC OXIDE SYNTHASE) IS AN ENDOTHELIAL CELL (EC)-SPECIFIC GENE PREDOMINANTLY EXPRESSED IN MEDIUM- TO LARGE-SIZED ARTERIES WHERE ECS EXPERIENCE ATHEROPROTECTIVE LAMINAR FLOW WITH HIGH SHEAR STRESS. DISTURBED FLOW WITH LOWER AVERAGE SHEAR STRESS DECREASES ENOS TRANSCRIPTION, WHICH LEADS TO THE DEVELOPMENT OF ATHEROSCLEROSIS, ESPECIALLY AT BIFURCATIONS AND CURVATURES OF ARTERIES. THIS PROTOTYPIC ARTERIAL EC GENE CONTAINS 2 DISTINCT FLOW-RESPONSIVE CIS-DNA ELEMENTS IN THE PROMOTER, THE SHEAR STRESS RESPONSE ELEMENT (SSRE) AND THE KLF (KRUPPEL-LIKE FACTOR) ELEMENT. PREVIOUS IN VITRO STUDIES SUGGESTED THEIR POSITIVE REGULATORY FUNCTIONS ON FLOW-INDUCED TRANSCRIPTION OF EC GENES INCLUDING ENOS. HOWEVER, THE IN VIVO FUNCTION OF THESE CIS-DNA ELEMENTS REMAINS UNKNOWN. METHODS: INSERTIONAL TRANSGENIC MICE WITH A MUTATION AT EACH FLOW-RESPONSIVE CIS-DNA ELEMENT WERE GENERATED USING A MURINE ENOS PROMOTER-BETA-GALACTOSIDASE REPORTER BY LINKER-SCANNING MUTAGENESIS AND COMPARED WITH EPISOMAL-BASED MUTATIONS IN VITRO. DNA METHYLATION AT THE ENOS PROXIMAL PROMOTER IN MOUSE ECS WAS ASSESSED BY BISULFITE SEQUENCING OR PYROSEQUENCING. RESULTS: WILD TYPE MICE WITH A FUNCTIONAL ENOS PROMOTER-REPORTER TRANSGENE EXHIBITED REDUCED ENDOTHELIAL REPORTER EXPRESSION IN THE ATHEROPRONE REGIONS OF DISTURBED FLOW (N=5). IT IS SURPRISING THAT THE SSRE MUTATION ABROGATED REPORTER EXPRESSION IN ECS AND WAS ASSOCIATED WITH ABERRANT HYPERMETHYLATION AT THE ENOS PROXIMAL PROMOTER (N=7). REPORTER GENE SILENCING WAS INDEPENDENT OF TRANSGENE COPY NUMBER AND INTEGRATION POSITION, INDICATING THAT THE SSRE IS A CRITICAL CIS-ELEMENT NECESSARY FOR ENOS TRANSCRIPTION IN VIVO. THE KLF MUTATION DEMONSTRATED AN INTEGRATION SITE-SPECIFIC DECREASE IN ENOS TRANSCRIPTION, AGAIN WITH MARKED PROMOTER METHYLATION (N=8), SUGGESTING THAT THE SSRE ALONE IS NOT SUFFICIENT FOR ENOS TRANSCRIPTION IN VIVO. IN WILD TYPE MICE, THE NATIVE ENOS PROMOTER WAS SIGNIFICANTLY HYPERMETHYLATED IN ECS FROM THE ATHEROPRONE REGIONS WHERE ENOS EXPRESSION WAS MARKEDLY REPRESSED BY CHRONIC DISTURBED FLOW, DEMONSTRATING THAT ENOS EXPRESSION IS REGULATED BY FLOW-DEPENDENT DNA METHYLATION THAT IS REGION-SPECIFIC IN THE ARTERIAL ENDOTHELIUM IN VIVO. CONCLUSIONS: WE REPORT, FOR THE FIRST TIME, THAT THE SSRE AND KLF ELEMENTS ARE CRITICAL FLOW SENSORS NECESSARY FOR A TRANSCRIPTIONALLY PERMISSIVE, HYPOMETHYLATED ENOS PROMOTER IN ECS UNDER CHRONIC SHEAR STRESS IN VIVO. MOREOVER, ENOS EXPRESSION IS REGULATED BY FLOW-DEPENDENT EPIGENETIC MECHANISMS, WHICH OFFERS NOVEL MECHANISTIC INSIGHT ON ENOS GENE REGULATION IN ATHEROGENESIS.	2021	
                                                                                                                                                                         
8  6456  29 THYMOSIN BETA4 PREVENTS OXIDATIVE STRESS, INFLAMMATION, AND FIBROSIS IN ETHANOL- AND LPS-INDUCED LIVER INJURY IN MICE. THYMOSIN BETA 4 (TBETA4), AN ACTIN-SEQUESTERING PROTEIN, IS INVOLVED IN TISSUE DEVELOPMENT AND REGENERATION. IT PREVENTS INFLAMMATION AND FIBROSIS IN SEVERAL TISSUES. WE INVESTIGATED THE ROLE OF TBETA4 IN CHRONIC ETHANOL- AND ACUTE LIPOPOLYSACCHARIDE- (LPS-) INDUCED MOUSE LIVER INJURY. C57BL/6 MICE WERE FED 5% ETHANOL IN LIQUID DIET FOR 4 WEEKS PLUS BINGE ETHANOL (5 G/KG, GAVAGE) WITH OR WITHOUT LPS (2 MG/KG, INTRAPERITONEAL) FOR 6 HOURS. TBETA4 (1 MG/KG, INTRAPERITONEAL) WAS ADMINISTERED FOR 1 WEEK. WE DEMONSTRATED THAT TBETA4 PREVENTED ETHANOL- AND LPS-MEDIATED INCREASE IN LIVER INJURY MARKERS AS WELL AS CHANGES IN LIVER PATHOLOGY. IT ALSO PREVENTED ETHANOL- AND LPS-MEDIATED INCREASE IN OXIDATIVE STRESS BY DECREASING ROS AND LIPID PEROXIDATION AND INCREASING THE ANTIOXIDANTS, REDUCED GLUTATHIONE AND MANGANESE-DEPENDENT SUPEROXIDE DISMUTASE. IT ALSO PREVENTED THE ACTIVATION OF NUCLEAR FACTOR KAPPA B BY BLOCKING THE PHOSPHORYLATION OF THE INHIBITORY PROTEIN, IKAPPAB, THEREBY PREVENTED PROINFLAMMATORY CYTOKINE PRODUCTION. MOREOVER, TBETA4 PREVENTED FIBROGENESIS BY SUPPRESSING THE EPIGENETIC REPRESSOR, METHYL-CPG-BINDING PROTEIN 2, THAT COORDINATELY REVERSED THE EXPRESSION OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-GAMMA AND DOWNREGULATED FIBROGENIC GENES, PLATELET-DERIVED GROWTH FACTOR-BETA RECEPTOR, ALPHA-SMOOTH MUSCLE ACTIN, COLLAGEN 1, AND FIBRONECTIN, RESULTING IN REDUCED FIBROSIS. OUR DATA SUGGEST THAT TBETA4 HAS ANTIOXIDANT, ANTI-INFLAMMATORY, AND ANTIFIBROTIC POTENTIAL DURING ALCOHOLIC LIVER INJURY.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
9  5860  38 SULFORAPHANE PREVENTS ANGIOTENSIN II-INDUCED CARDIOMYOPATHY BY ACTIVATION OF NRF2 THROUGH EPIGENETIC MODIFICATION. NUCLEAR FACTOR ERYTHROID 2-RELATED FACTOR (NRF2) IS AN IMPORTANT REGULATOR OF CELLULAR ANTIOXIDANT DEFENCE. WE PREVIOUSLY SHOWED THAT SFN PREVENTED ANG II-INDUCED CARDIAC DAMAGE VIA ACTIVATION OF NRF2. HOWEVER, THE UNDERLYING MECHANISM OF SFN'S PERSISTENT CARDIAC PROTECTION REMAINS UNCLEAR. THIS STUDY AIMED TO EXPLORE THE POTENTIAL OF SFN IN ACTIVATING CARDIAC NRF2 THROUGH EPIGENETIC MECHANISMS. WILD-TYPE MICE WERE INJECTED SUBCUTANEOUSLY WITH ANG II, WITH OR WITHOUT SFN. ADMINISTRATION OF CHRONIC ANG II-INDUCED CARDIAC INFLAMMATORY FACTOR EXPRESSION, OXIDATIVE DAMAGE, FIBROSIS AND CARDIAC REMODELLING AND DYSFUNCTION, ALL OF WHICH WERE EFFECTIVELY IMPROVED BY SFN TREATMENT, COUPLED WITH AN UP-REGULATION OF NRF2 AND DOWNSTREAM GENES. BISULFITE GENOME SEQUENCING AND CHROMATIN IMMUNOPRECIPITATION (CHIP) WERE PERFORMED TO DETECT THE METHYLATION LEVEL OF THE FIRST 15 CPGS AND HISTONE H3 ACETYLATION (AC-H3) STATUS IN THE NRF2 PROMOTER REGION, RESPECTIVELY. THE RESULTS SHOWED THAT SFN REDUCED ANG II-INDUCED CPG HYPERMETHYLATION AND PROMOTED AC-H3 ACCUMULATION IN THE NRF2 PROMOTER REGION, ACCOMPANIED BY THE INHIBITION OF GLOBAL DNMT AND HDAC ACTIVITY, AND A DECREASED PROTEIN EXPRESSION OF KEY DNMT AND HDAC ENZYMES. TAKEN TOGETHER, SFN EXERTS ITS CARDIOPROTECTIVE EFFECT THROUGH EPIGENETIC MODIFICATION OF NRF2, WHICH MAY PARTIALLY CONTRIBUTE TO LONG-TERM ACTIVATION OF CARDIAC NRF2.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
10  4303  36 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
11  3792  31 INTERLEUKIN-1BETA INCREASES THE RISK OF GASTRIC CANCER THROUGH INDUCTION OF ABERRANT DNA METHYLATION IN A MOUSE MODEL. INTERLEUKIN-1BETA (IL-1BETA) HAS A SIGNIFICANT ROLE IN CHRONIC GASTRIC INFLAMMATION AND MANIFESTATIONS OF GASTRIC DISEASES. THE PRESENT STUDY AIMED TO ELUCIDATE THE SPECIFIC ROLE OF IL-1BETA IN INDUCTION OF DNA METHYLATION USING IL-1 RECEPTOR TYPE 1 KNOCKOUT (IL-1R1(-)/(-)) MICE. IN THE PRESENT STUDY, WILD-TYPE (WT) AND IL-1R1(-)/(-) MICE WERE INJECTED WITH IL-1BETA (5 MICROG/KG/DAY). SERUM LEVELS OF IL-1BETA, INTERLEUKIN-6 (IL-6) AND NITRIC OXIDE (NO) WERE MEASURED BY ENZYME-LINKED IMMUNOSORBENT OR NO ASSAYS. E-CADHERIN (E-CAD) METHYLATION STATUS AND MESSENGER (M)RNA EXPRESSION OF IL-1BETA, IL-6, E-CAD AND INDUCIBLE NITRIC OXIDE SYNTHASE (INOS) WERE ANALYZED. RESULTS FROM THE PRESENT STUDY INDICATED SIGNIFICANTLY HIGHER IL-1BETA MRNA EXPRESSION (P<0.001) IN WT MICE COMPARED WITH IL-1R1(-)/(-) MICE. IL-1BETA AND IL-6 RELEASE WAS SIGNIFICANTLY INCREASED IN TREATED WT MICE COMPARED WITH IL-1R1(-)/(-) MICE AT 1 H, 4 H AND 8 H (ALL P<0.005). IL-1BETA RELEASE WAS ONLY DETECTED IN WT MICE FOLLOWING A SECOND DOSE MEASURED AT DAY 3, WEEK 1 AND WEEK 2 WHEN COMPARED WITH IL-1R1(-)/(-) MICE. PROMOTER METHYLATION OF E-CAD AND A DECREASE IN GENE EXPRESSION WAS OBSERVED IN TREATED WT MICE. MRNA EXPRESSION OF INOS IN WT MICE WAS SIGNIFICANTLY INCREASED AT WEEK 1 COMPARED WITH IL-1R1(-)/(-) MICE (P=0.0411). FURTHERMORE, A SIGNIFICANTLY INCREASED LEVEL OF NO PRODUCTION WAS OBSERVED IN TREATED WT MICE (P<0.005 AT 8 H AND WEEK 1; P<0.001 AT 4 H AND DAY 3) WHEN COMPARED WITH IL-1R1(-)/(-) MICE. THE PRESENT RESULTS INDICATED THAT IL-1BETA WAS ABLE TO DIRECTLY INDUCE DNA METHYLATION, WHICH MAY LINK INFLAMMATION-INDUCED EPIGENETIC CHANGES AND THE DEVELOPMENT OF GASTRIC DISEASES.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
12  2950  24 GENETIC AND EPIGENETIC DAMAGE INDUCED BY REACTIVE NITROGEN SPECIES: IMPLICATIONS IN CARCINOGENESIS. CHRONIC INFECTION AND INFLAMMATION ARE RECOGNIZED RISK FACTORS FOR HUMAN CANCER AT VARIOUS SITES. INFECTION AND INFLAMMATION CAN ACTIVATE AND INDUCE A VARIETY OF OXIDANT-GENERATING ENZYMES, INCLUDING NADPH OXIDASE AND INDUCIBLE NITRIC OXIDE SYNTHASE. REACTIVE OXYGEN AND NITROGEN SPECIES PRODUCED BY SUCH ENZYMES REACT WITH EACH OTHER TO GENERATE NEW AND MORE POTENT REACTIVE SPECIES. THESE OXIDANTS NOT ONLY CAN DAMAGE DNA AND INDUCE MUTATIONS, BUT ALSO CAN ACTIVATE ONCOGENE PRODUCTS AND/OR INACTIVATE TUMOR-SUPPRESSOR PROTEINS, THUS CONTRIBUTING TO MOST PROCESSES OF CARCINOGENESIS. APPROPRIATE TREATMENT OF INFLAMMATION SHOULD BE FURTHER EXPLORED FOR CHEMOPREVENTION OF HUMAN CANCERS, ESPECIALLY THOSE ASSOCIATED WITH CHRONIC INFLAMMATION.	2003	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
13  4841  29 ONE WEEK, BUT NOT 12 HOURS, OF CAST IMMOBILIZATION ALTERS PROMOTOR DNA METHYLATION PATTERNS IN THE NNOS GENE IN MOUSE SKELETAL MUSCLE. KEY POINTS: DNA METHYLATION MAY PLAY AN IMPORTANT ROLE IN REGULATING GENE EXPRESSION IN SKELETAL MUSCLE TO ADAPT TO PHYSICAL ACTIVITY AND INACTIVITY. NEURONAL NITRIC OXIDE SYNTHASE (NNOS) IN SKELETAL MUSCLE IS A KEY REGULATOR OF SKELETAL MUSCLE MASS; HOWEVER, IT IS UNCLEAR WHETHER NNOS EXPRESSION IS REGULATED BY DNA METHYLATION. WE FOUND THAT 1 WEEK OF CAST IMMOBILIZATION INCREASED NNOS DNA METHYLATION LEVELS AND DOWNREGULATED NNOS GENE EXPRESSION IN ATROPHIC SLOW-TWITCH SOLEUS MUSCLE FROM THE MOUSE LEG. THESE CHANGES WERE NOT DETECTED IN NON-ATROPHIC FAST-TWITCH EXTENSOR DIGITORUM LONGUS MUSCLE. TWELVE HOURS OF CAST IMMOBILIZATION DECREASED NNOS GENE EXPRESSION, WHEREAS NNOS DNA METHYLATION LEVELS WERE UNCHANGED, SUGGESTING THAT DOWNREGULATION OF NNOS GENE EXPRESSION BY SHORT-TERM MUSCLE INACTIVITY IS INDEPENDENT OF THE DNA METHYLATION PATTERN. THESE FINDINGS CONTRIBUTE TO A BETTER UNDERSTANDING OF THE MAINTENANCE OF SKELETAL MUSCLE MASS AND PREVENTION OF MUSCLE ATROPHY BY EPIGENETIC MECHANISMS VIA THE NNOS/NO PATHWAY. ABSTRACT: DNA METHYLATION IS A MECHANISM THAT CONTROLS GENE EXPRESSION IN SKELETAL MUSCLE UNDER VARIOUS ENVIRONMENTAL STIMULI, SUCH AS PHYSICAL ACTIVITY AND INACTIVITY. NEURONAL NITRIC OXIDE SYNTHASE (NNOS) REGULATES MUSCLE ATROPHY IN SKELETAL MUSCLE. HOWEVER, THE MECHANISMS REGULATING NNOS EXPRESSION IN ATROPHIC MUSCLE REMAIN UNCLEAR. WE HYPOTHESIZED THAT NNOS EXPRESSION IN ATROPHIC MUSCLE IS REGULATED BY DNA METHYLATION OF THE NNOS PROMOTOR IN SOLEUS (SOL; SLOW-TWITCH FIBRE DOMINANT) AND EXTENSOR DIGITORUM LONGUS (EDL; FAST-TWITCH FIBRE DOMINANT) MUSCLES. ONE WEEK OF CAST IMMOBILIZATION INDUCED SIGNIFICANT MUSCLE ATROPHY IN SOL BUT NOT IN EDL. WE SHOWED THAT 1 WEEK OF CAST IMMOBILIZATION INCREASED NNOS DNA METHYLATION LEVELS IN SOL, ALTHOUGH ONLY A MINOR CHANGE WAS DETECTED IN EDL. CONSISTENT WITH THE INCREASED DNA METHYLATION LEVELS IN ATROPHIC SOL, THE GENE EXPRESSION LEVELS OF TOTAL NNOS AND NNOSMICRO (I.E. THE MAJOR SPLICING VARIANT OF NNOS IN SKELETAL MUSCLE) DECREASED. THE ABUNDANCE OF THE NNOS PROTEIN AND CELL MEMBRANE (ESPECIALLY TYPE IIA FIBRE) IMMUNOREACTIVITY ALSO DECREASED IN ATROPHIC SOL. THESE CHANGES WERE NOT OBSERVED IN EDL AFTER 1 WEEK OF CAST IMMOBILIZATION. FURTHERMORE, DESPITE THE LACK OF SIGNIFICANT ATROPHY, 12 H OF CAST IMMOBILIZATION DECREASED GENE EXPRESSION LEVELS OF TOTAL NNOS AND NNOSMICRO IN SOL. HOWEVER, NO ASSOCIATION WAS DETECTED BETWEEN NNOS DNA METHYLATION AND GENE EXPRESSION. THE EXPRESSION OF THE NNOSBETA GENE, ANOTHER SPLICING VARIANT OF NNOS, IN EDL WAS UNCHANGED BY CAST IMMOBILIZATION, WHEREAS ITS EXPRESSION WAS NOT DETECTED IN SOL. WE CONCLUDED THAT CHRONIC ADAPTATION OF NNOS GENE EXPRESSION IN CAST IMMOBILIZED MUSCLE MAY INVOLVE NNOS DNA METHYLATION.	2019	

14  5868  34 SUPPRESSIVE EFFECTS OF METFORMIN ON T-HELPER 1-RELATED CHEMOKINES EXPRESSION IN THE HUMAN MONOCYTIC LEUKEMIA CELL LINE THP-1. PURPOSE OF THE STUDY: TYPE 1 AND TYPE 2 DIABETES MELLITUS (DM) ARE CHRONIC T-CELL-MEDIATED INFLAMMATORY DISEASES. METFORMIN IS A WIDELY USED DRUG FOR TYPE 2 DM THAT REDUCES THE NEED FOR INSULIN IN TYPE 1 DM. HOWEVER, WHETHER METFORMIN HAS AN ANTI-INFLAMMATORY EFFECT FOR TREATING DM IS UNKNOWN. WE INVESTIGATED THE ANTI-INFLAMMATORY MECHANISM OF METFORMIN IN THE HUMAN MONOCYTIC LEUKEMIA CELL LINE THP-1. MATERIALS AND METHODS: THE HUMAN MONOCYTIC LEUKEMIA CELL LINE THP-1 WAS PRETREATED WITH METFORMIN AND STIMULATED WITH LIPOPOLYSACCHARIDE (LPS). THE PRODUCTION OF T-HELPER (TH)-1-RELATED CHEMOKINES INCLUDING INTERFERON-GAMMA-INDUCED PROTEIN-10 (IP-10) AND MONOCYTE CHEMOATTRACTANT PROTEIN-1 (MCP-1), TH2-RELATED CHEMOKINE MACROPHAGE-DERIVED CHEMOKINE, AND THE PROINFLAMMATORY CHEMOKINE TUMOR NECROSIS FACTOR-ALPHA WAS MEASURED USING ENZYME-LINKED IMMUNOSORBENT ASSAY. INTRACELLULAR SIGNALING PATHWAYS WERE INVESTIGATED USING WESTERN BLOT ANALYSIS AND CHROMATIN IMMUNOPRECIPITATION ASSAY. RESULTS: METFORMIN SUPPRESSED LPS-INDUCED IP-10 AND MCP-1 PRODUCTION AS WELL AS LPS-INDUCED PHOSPHORYLATION OF C-JUN N-TERMINAL KINASE (JNK), P38, EXTRACELLULAR SIGNAL-REGULATED KINASE (ERK), AND NUCLEAR FACTOR-KAPPA B (NF-KAPPAB). MOREOVER, METFORMIN SUPPRESSED LPS-INDUCED ACETYLATION OF HISTONES H3 AND H4 AT THE IP-10 PROMOTER. CONCLUSIONS: METFORMIN SUPPRESSED THE PRODUCTION OF TH1-RELATED CHEMOKINES IP-10 AND MCP-1 IN THP-1 CELLS. SUPPRESSIVE EFFECTS OF METFORMIN ON IP-10 PRODUCTION MIGHT BE ATTRIBUTED AT LEAST PARTIALLY TO THE JNK, P38, ERK, AND NF-KAPPAB PATHWAYS AS WELL AS TO EPIGENETIC REGULATION THROUGH THE ACETYLATION OF HISTONES H3 AND H4. THESE RESULTS INDICATED THE THERAPEUTIC ANTI-INFLAMMATORY POTENTIAL OF METFORMIN.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
15   169  31 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
16  6166  31 THE GLUTATHIONE SYSTEM: A NEW DRUG TARGET IN NEUROIMMUNE DISORDERS. GLUTATHIONE (GSH) HAS A CRUCIAL ROLE IN CELLULAR SIGNALING AND ANTIOXIDANT DEFENSES EITHER BY REACTING DIRECTLY WITH REACTIVE OXYGEN OR NITROGEN SPECIES OR BY ACTING AS AN ESSENTIAL COFACTOR FOR GSH S-TRANSFERASES AND GLUTATHIONE PEROXIDASES. GSH ACTING IN CONCERT WITH ITS DEPENDENT ENZYMES, KNOWN AS THE GLUTATHIONE SYSTEM, IS RESPONSIBLE FOR THE DETOXIFICATION OF REACTIVE OXYGEN AND NITROGEN SPECIES (ROS/RNS) AND ELECTROPHILES PRODUCED BY XENOBIOTICS. ADEQUATE LEVELS OF GSH ARE ESSENTIAL FOR THE OPTIMAL FUNCTIONING OF THE IMMUNE SYSTEM IN GENERAL AND T CELL ACTIVATION AND DIFFERENTIATION IN PARTICULAR. GSH IS A UBIQUITOUS REGULATOR OF THE CELL CYCLE PER SE. GSH ALSO HAS CRUCIAL FUNCTIONS IN THE BRAIN AS AN ANTIOXIDANT, NEUROMODULATOR, NEUROTRANSMITTER, AND ENABLER OF NEURON SURVIVAL. DEPLETION OF GSH LEADS TO EXACERBATION OF DAMAGE BY OXIDATIVE AND NITROSATIVE STRESS; HYPERNITROSYLATION; INCREASED LEVELS OF PROINFLAMMATORY MEDIATORS AND INFLAMMATORY POTENTIAL; DYSFUNCTIONS OF INTRACELLULAR SIGNALING NETWORKS, E.G., P53, NUCLEAR FACTOR-KAPPAB, AND JANUS KINASES; DECREASED CELL PROLIFERATION AND DNA SYNTHESIS; INACTIVATION OF COMPLEX I OF THE ELECTRON TRANSPORT CHAIN; ACTIVATION OF CYTOCHROME C AND THE APOPTOTIC MACHINERY; BLOCKADE OF THE METHIONINE CYCLE; AND COMPROMISED EPIGENETIC REGULATION OF GENE EXPRESSION. AS SUCH, GSH DEPLETION HAS MARKED CONSEQUENCES FOR THE HOMEOSTATIC CONTROL OF THE IMMUNE SYSTEM, OXIDATIVE AND NITROSATIVE STRESS (O&NS) PATHWAYS, REGULATION OF ENERGY PRODUCTION, AND MITOCHONDRIAL SURVIVAL AS WELL. GSH DEPLETION AND CONCOMITANT INCREASE IN O&NS AND MITOCHONDRIAL DYSFUNCTIONS PLAY A ROLE IN THE PATHOPHYSIOLOGY OF DIVERSE NEUROIMMUNE DISORDERS, INCLUDING DEPRESSION, MYALGIC ENCEPHALOMYELITIS/CHRONIC FATIGUE SYNDROME AND PARKINSON'S DISEASE, SUGGESTING THAT DEPLETED GSH IS AN INTEGRAL PART OF THESE DISEASES. THERAPEUTICAL INTERVENTIONS THAT AIM TO INCREASE GSH CONCENTRATIONS IN VIVO INCLUDE N-ACETYL CYSTEINE; NRF-2 ACTIVATION VIA HYPERBARIC OXYGEN THERAPY; DIMETHYL FUMARATE; PHYTOCHEMICALS, INCLUDING CURCUMIN, RESVERATROL, AND CINNAMON; AND FOLATE SUPPLEMENTATION.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
17  4044  28 MACROPHAGES IN OXIDATIVE STRESS AND MODELS TO EVALUATE THE ANTIOXIDANT FUNCTION OF DIETARY NATURAL COMPOUNDS. ANTIOXIDANT TESTING OF NATURAL PRODUCTS HAS ATTRACTED INCREASING INTEREST IN RECENT YEARS, MAINLY DUE TO THE FACT THAT AN ANTIOXIDANT-RICH DIET MIGHT PROVIDE HEALTH BENEFITS. ACTIVATED MACROPHAGES ARE A MAJOR SOURCE OF REACTIVE OXYGEN SPECIES, REACTIVE NITROGEN SPECIES, AND PEROXYNITRITE GENERATED THROUGH THE SO-CALLED RESPIRATORY BURST. CONSTITUTIVELY RELEASED PROINFLAMMATORY CYTOKINE, ESPECIALLY TUMOR NECROSIS FACTOR-ALPHA, TRIGGERS NUCLEAR FACTOR-KAPPAB, AND ACTIVATOR PROTEIN-1 TRANSLOCATION LEADING TO THE OVER PRODUCTION OF REACTIVE OXYGEN SPECIES AND REACTIVE NITROGEN SPECIES IN MACROPHAGES. ACTIVATION OF TRANSCRIPTION FACTORS IN THE LONG-LIVED TISSUE-RESIDENT MACROPHAGES AND/OR MONOCYTE-DERIVED MACROPHAGES, TRIGGER EPIGENETIC MODIFICATIONS LEADING TO THE PATHOGENESIS OF CHRONIC DISEASES. NUTRACEUTICALS INCLUDING LIPID RAFT STRUCTURE DISRUPTION AGENT, CHOLESTEROL DEPLETION AGENT, FARNESYLTRANSFERASE INHIBITOR, NUCLEAR FACTOR-KAPPAB BLOCKER (ALPHA,BETA-UNSATURATED CARBONYL COMPOUNDS), GLUCOCORTICOID RECEPTOR AGONIST, AND PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-GAMMA AGONIST HAVE LONG BEEN USED TO INACTIVE MACROPHAGE. THE INHIBITION EFFECTS ON THE FORMATION OF NITRIC OXIDE, SUPEROXIDE, AND NITRITE PEROXIDE MAY BE RESPONSIBLE FOR THE ANTI-INFLAMMATORY FUNCTIONALITIES. ACTIVATED MACROPHAGE MODELS COULD BE USED TO IDENTIFY THE ACTIVE COMPONENTS FOR FUNCTIONAL DIETS DEVELOPMENT THROUGH A MULTIPLE TARGETS STRATEGY.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
18  5760  38 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
19  1945  37 EPIGALLOCATECHIN-3-GALLATE, A HISTONE ACETYLTRANSFERASE INHIBITOR, INHIBITS EBV-INDUCED B LYMPHOCYTE TRANSFORMATION VIA SUPPRESSION OF RELA ACETYLATION. BECAUSE THE P300/CBP-MEDIATED HYPERACETYLATION OF RELA (P65) IS CRITICAL FOR NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) ACTIVATION, THE ATTENUATION OF P65 ACETYLATION IS A POTENTIAL MOLECULAR TARGET FOR THE PREVENTION OF CHRONIC INFLAMMATION. DURING OUR ONGOING SCREENING STUDY TO IDENTIFY NATURAL COMPOUNDS WITH HISTONE ACETYLTRANSFERASE INHIBITOR (HATI) ACTIVITY, WE IDENTIFIED EPIGALLOCATECHIN-3-GALLATE (EGCG) AS A NOVEL HATI WITH GLOBAL SPECIFICITY FOR THE MAJORITY OF HAT ENZYMES BUT WITH NO ACTIVITY TOWARD EPIGENETIC ENZYMES INCLUDING HDAC, SIRT1, AND HMTASE. AT A DOSE OF 100 MICROMOL/L, EGCG ABROGATES P300-INDUCED P65 ACETYLATION IN VITRO AND IN VIVO, INCREASES THE LEVEL OF CYTOSOLIC IKAPPABALPHA, AND SUPPRESSES TUMOR NECROSIS FACTOR ALPHA (TNFALPHA)-INDUCED NF-KAPPAB ACTIVATION. WE ALSO SHOWED THAT EGCG PREVENTS TNFALPHA-INDUCED P65 TRANSLOCATION TO THE NUCLEUS, CONFIRMING THAT HYPERACETYLATION IS CRITICAL FOR NF-KAPPAB TRANSLOCATION AS WELL AS ACTIVITY. FURTHERMORE, EGCG TREATMENT INHIBITED THE ACETYLATION OF P65 AND THE EXPRESSION OF NF-KAPPAB TARGET GENES IN RESPONSE TO DIVERSE STIMULI. FINALLY, EGCG REDUCED THE BINDING OF P300 TO THE PROMOTER REGION OF INTERLEUKIN-6 GENE WITH AN INCREASED RECRUITMENT OF HDAC3, WHICH HIGHLIGHTS THE IMPORTANCE OF THE BALANCE BETWEEN HATS AND HISTONE DEACETYLASES IN THE NF-KAPPAB-MEDIATED INFLAMMATORY SIGNALING PATHWAY. IMPORTANTLY, EGCG AT 50 MICROMOL/L DOSE COMPLETELY BLOCKS EBV INFECTION-INDUCED CYTOKINE EXPRESSION AND SUBSEQUENTLY THE EBV-INDUCED B LYMPHOCYTE TRANSFORMATION. THESE RESULTS SHOW THE CRUCIAL ROLE OF ACETYLATION IN THE DEVELOPMENT OF INFLAMMATORY-RELATED DISEASES.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
20  1826  36 EFFECTS OF HISTONE DEACETYLASE INHIBITOR ON EXTRACELLULAR MATRIX PRODUCTION IN HUMAN NASAL POLYP ORGAN CULTURES. BACKGROUND: NASAL POLYPOSIS IS ASSOCIATED WITH A CHRONIC INFLAMMATORY CONDITION OF THE SINONASAL MUCOSA AND INVOLVES MYOFIBROBLAST DIFFERENTIATION AND EXTRACELLULAR MATRIX (ECM) ACCUMULATION. EPIGENETIC MODULATION BY HISTONE DEACETYLASE (HDAC) INHIBITORS INCLUDING TRICHOSTATIN A (TSA) HAS BEEN REPORTED TO HAVE INHIBITORY EFFECTS ON MYOFIBROBLAST DIFFERENTIATION IN LUNG AND RENAL FIBROBLASTS. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE THE INHIBITORY EFFECT OF TSA ON MYOFIBROBLAST DIFFERENTIATION AND ECM PRODUCTION IN NASAL POLYP ORGAN CULTURES. METHODS: NASAL POLYP TISSUES FROM 18 PATIENTS WERE ACQUIRED DURING ENDOSCOPIC SINUS SURGERY. AFTER ORGAN CULTURE, NASAL POLYPS WERE STIMULATED WITH TGF-BETA1 AND THEN TREATED WITH TSA. ALPHA-SMOOTH MUSCLE ACTIN (ALPHA-SMA), FIBRONECTIN, AND COLLAGEN TYPE I EXPRESSION LEVELS WERE EXAMINED BY REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION (PCR), REAL-TIME PCR, WESTERN BLOT, AND IMMUNOFLUORESCENT STAINING. HDAC2, HDAC4, AND ACETYLATED H4 EXPRESSION LEVELS WERE ASSAYED BY WESTERN BLOT. CYTOTOXICITY WAS ANALYZED BY THE TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE BIOTIN-DUTP NICK END LABELING ASSAY. RESULTS: THE EXPRESSION LEVELS OF ALPHA-SMA, FIBRONECTIN, AND COLLAGEN TYPE 1 WERE INCREASED IN NASAL POLYP AFTER TRANSFORMING GROWTH FACTOR (TGF) BETA1 TREATMENT. TSA-INHIBITED TGF-BETA1 INDUCED THESE GENE AND PROTEIN EXPRESSION LEVELS. FURTHERMORE, TSA SUPPRESSED PROTEIN EXPRESSION LEVELS OF HDAC2 AND HDAC4. HOWEVER, TSA INDUCED HYPERACETYLATION OF HISTONES H4. TREATMENT WITH TGF-BETA1 WITH OR WITHOUT TSA DID NOT HAVE CYTOTOXIC EFFECT. CONCLUSION: THESE FINDINGS PROVIDE NOVEL INSIGHTS INTO THE EPIGENETIC REGULATION IN MYOFIBROBLAST DIFFERENTIATION AND ECM PRODUCTION OF NASAL POLYP. TSA COULD BE A CANDIDATE OF A THERAPEUTIC AGENT FOR REVERSING THE TGF-BETA1-INDUCED ECM SYNTHESIS THAT LEADS TO NASAL POLYP DEVELOPMENT.	2013