1  2197 88 EPIGENETIC MODIFICATION OF BDNF MEDIATES NEUROPATHIC PAIN VIA MIR-30A-3P/EP300 AXIS IN CCI RATS. RECENT INVESTIGATION OF MICRORNAS ON CHRONIC PAIN HAS DEVELOPED A BREAKTHROUGH IN NEUROPATHIC PAIN MANAGEMENT. IN THE PRESENT STUDY, DECREASED EXPRESSION OF MIR-30A-3P WAS REPORTED USING QRT-PCR ANALYSIS AND LOSS OF MIR-30A-3P PROMOTED NEUROPATHIC PAIN PROGRESSION IN SCIATIC NERVE CHRONIC CONSTRICTIVE INJURY RATS THROUGH DETERMINING THE PAIN THRESHOLD. WE PREDICTED MIR-30A-3P COULD TARGET E-CADHERIN TRANSCRIPTIONAL ACTIVATOR (EP300) VIA BIOINFORMATICS ANALYSIS. MEANWHILE, WE FOUND THAT BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) IS INVOLVED IN NEUROPATHIC PAIN. HERE, WE EXHIBITED THAT EP300 EPIGENETICALLY UP-REGULATED BDNF VIA ENHANCING ACETYLATED HISTONE H3 AND H4 ON THE PROMOTER. FOR ANOTHER, MIR-30A-3P WAS ABLE TO MODIFY THE LEVEL OF BDNF AND ACETYLATED HISTONE H3 AND H4. LOSS OF MIR-30A-3P ENHANCED EP300 AND BDNF COLOCALIZATION IN CCI RATS. SUBSEQUENTLY, IT WAS SHOWN THAT INCREASED EP300 INDUCED NEUROPATHIC PAIN BY AN ENHANCEMENT OF NEURONAL BDNF LEVEL IN VIVO. TO SUM UP, IT WAS REVEALED THAT EPIGENETIC MODIFICATION OF BDNF PROMOTED NEUROPATHIC PAIN VIA EP300 INDUCED BY MIR-30A-3P IN CCI RATS.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
2  4546 26 MUTANT P53 REGULATES ENHANCER-ASSOCIATED H3K4 MONOMETHYLATION THROUGH INTERACTIONS WITH THE METHYLTRANSFERASE MLL4. MONOMETHYLATION OF HISTONE H3 LYSINE 4 (H3K4ME1) IS ENRICHED AT ENHANCERS THAT ARE PRIMED FOR ACTIVATION AND THE LEVELS OF THIS HISTONE MARK ARE FREQUENTLY ALTERED IN VARIOUS HUMAN CANCERS. YET, HOW ALTERATIONS IN H3K4ME1 ARE ESTABLISHED AND THE CONSEQUENCES OF THESE EPIGENETIC CHANGES IN TUMORIGENESIS ARE NOT WELL UNDERSTOOD. USING CHIP-SEQ IN HUMAN COLON CANCER CELLS, WE DEMONSTRATE THAT MUTANT P53 DEPLETION RESULTS IN DECREASED H3K4ME1 LEVELS AT ACTIVE ENHANCERS THAT REVEAL A STRIKING COLOCALIZATION OF MUTANT P53 AND THE H3K4 MONOMETHYLTRANSFERASE MLL4 FOLLOWING CHRONIC TUMOR NECROSIS FACTOR ALPHA (TNFALPHA) SIGNALING. WE FURTHER REVEAL THAT MUTANT P53 FORMS PHYSIOLOGICAL ASSOCIATIONS AND DIRECT INTERACTIONS WITH MLL4 AND PROMOTES THE ENHANCER BINDING OF MLL4, WHICH IS REQUIRED FOR TNFALPHA-INDUCIBLE H3K4ME1 AND HISTONE H3 LYSINE 27 ACETYLATION (H3K27AC) LEVELS, ENHANCER-DERIVED TRANSCRIPT (ERNA) SYNTHESIS, AND MUTANT P53-DEPENDENT TARGET GENE ACTIVATION. COMPLEMENTARY IN VITRO STUDIES WITH RECOMBINANT CHROMATIN AND PURIFIED PROTEINS DEMONSTRATE THAT BINDING OF THE MLL3/4 COMPLEX AND H3K4ME1 DEPOSITION IS ENHANCED BY MUTANT P53 AND P300-MEDIATED ACETYLATION, WHICH IN TURN REFLECTS A MLL3/4-DEPENDENT ENHANCEMENT OF MUTANT P53 AND P300-DEPENDENT TRANSCRIPTIONAL ACTIVATION. COLLECTIVELY, OUR FINDINGS ESTABLISH A MECHANISM IN WHICH MUTANT P53 COOPERATES WITH MLL4 TO REGULATE ABERRANT ENHANCER ACTIVITY AND TUMOR-PROMOTING GENE EXPRESSION IN RESPONSE TO CHRONIC IMMUNE SIGNALING.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
3   687 32 BRAINSTEM BRAIN-DERIVED NEUROTROPHIC FACTOR SIGNALING IS REQUIRED FOR HISTONE DEACETYLASE INHIBITOR-INDUCED PAIN RELIEF. OUR PREVIOUS STUDY DEMONSTRATED THAT PERSISTENT PAIN CAN EPIGENETICALLY SUPPRESS THE TRANSCRIPTION OF GAD2 [ENCODING GLUTAMIC ACID DECARBOXYLASE 65 (GAD65)] AND CONSEQUENTLY IMPAIR THE INHIBITORY FUNCTION OF GABAERGIC SYNAPSES IN CENTRAL PAIN-MODULATING NEURONS. THIS CONTRIBUTES TO THE DEVELOPMENT OF PERSISTENT PAIN SENSITIZATION. HISTONE DEACETYLASE (HDAC) INHIBITORS INCREASED GAD65 ACTIVITY CONSIDERABLY, RESTORED GABA SYNAPTIC FUNCTION, AND RENDERED SENSITIZED PAIN BEHAVIOR LESS PRONOUNCED. HOWEVER, THE MOLECULAR MECHANISMS BY WHICH HDAC REGULATES GABAERGIC TRANSMISSION THROUGH GAD65 UNDER PAIN CONDITIONS ARE UNKNOWN. THIS WORK SHOWED THAT HDAC INHIBITOR-INDUCED INCREASES IN COLOCALIZATION OF GAD65 AND SYNAPTIC PROTEIN SYNAPSIN I ON THE PRESYNAPTIC AXON TERMINALS OF THE NUCLEUS RAPHE MAGNUS (NRM) WERE BLOCKED BY A TRKB RECEPTOR ANTAGONIST K252A [(9S,10R,12R)-2,3,9,10,11,12-HEXAHYDRO-10-HYDROXY-9-METHYL-1-OXO-9,12-EPOXY-1H-DIINDOLO[1,2,3-FG:3',2',1'-KL]PYRROLO[3,4-I][1,6]BENZODIAZOCINE-10-CARBOXYLIC ACID METHYL ESTER], INDICATING THAT BDNF-TRKB SIGNALING MAY BE REQUIRED IN GAD65 MODULATION OF GABA SYNAPTIC FUNCTION. AT THE BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) PROMOTER, HDAC INHIBITORS INDUCED SIGNIFICANT INCREASES IN H3 HYPERACETYLATION, CONSISTENT WITH THE INCREASE IN BDNF MRNA AND TOTAL PROTEINS. ALTHOUGH EXOGENOUS BDNF FACILITATED GABA MINIATURE INHIBITORY POSTSYNAPTIC CURRENTS AND GAD65 ACCUMULATION IN NRM NEURONAL SYNAPSES IN NORMAL RATS, IT FAILED TO DO SO IN ANIMALS SUBJECTED TO PERSISTENT INFLAMMATION. IN ADDITION, BLOCKADE OF THE TRKB RECEPTOR WITH K252A HAS NO EFFECT ON MINIATURE INHIBITORY POSTSYNAPTIC CURRENTS AND SYNAPTIC GAD65 ACCUMULATION UNDER NORMAL CONDITIONS. IN ADDITION, THE ANALGESIC EFFECTS OF HDAC INHIBITORS ON BEHAVIOR WERE BLOCKED BY NRM INFUSION OF K252A. THESE FINDINGS SUGGEST THAT BDNF-TRKB SIGNALING IS REQUIRED FOR DRUGS THAT REVERSE THE EPIGENETIC EFFECTS OF CHRONIC PAIN AT THE GENE LEVEL, SUCH AS HDAC INHIBITORS.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
4  3420 14 HUMAN LUNG DNA METHYLATION QUANTITATIVE TRAIT LOCI COLOCALIZE WITH CHRONIC OBSTRUCTIVE PULMONARY DISEASE GENOME-WIDE ASSOCIATION LOCI. RATIONALE: AS THE THIRD LEADING CAUSE OF DEATH IN THE UNITED STATES, THE IMPACT OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) MAKES IDENTIFICATION OF ITS MOLECULAR MECHANISMS OF GREAT IMPORTANCE. GENOME-WIDE ASSOCIATION STUDIES (GWASS) HAVE IDENTIFIED MULTIPLE GENOMIC REGIONS ASSOCIATED WITH COPD. HOWEVER, GENETIC VARIATION ONLY EXPLAINS A SMALL FRACTION OF THE SUSCEPTIBILITY TO COPD, AND SUB-GENOME-WIDE SIGNIFICANT LOCI MAY PLAY A ROLE IN PATHOGENESIS. OBJECTIVES: REGULATORY ANNOTATION WITH EPIGENETIC EVIDENCE MAY GIVE PRIORITY FOR FURTHER INVESTIGATION, PARTICULARLY FOR GWAS ASSOCIATIONS IN NONCODING REGIONS. WE PERFORMED INTEGRATIVE GENOMICS ANALYSES USING DNA METHYLATION PROFILING AND GENOME-WIDE SNP GENOTYPING FROM LUNG TISSUE SAMPLES FROM 90 SUBJECTS WITH COPD AND 36 CONTROL SUBJECTS. METHODS: WE PERFORMED METHYLATION QUANTITATIVE TRAIT LOCI (MQTL) ANALYSES, TESTING FOR SNPS ASSOCIATED WITH PERCENT DNA METHYLATION AND ASSESSED THE COLOCALIZATION OF THESE RESULTS WITH PREVIOUS COPD GWAS FINDINGS USING BAYESIAN METHODS IN THE R PACKAGE COLOC TO HIGHLIGHT POTENTIAL REGULATORY FEATURES OF THE LOCI. MEASUREMENTS AND MAIN RESULTS: WE IDENTIFIED 942,068 UNIQUE SNPS AND 33,996 UNIQUE CPG SITES AMONG THE SIGNIFICANT (5% FALSE DISCOVERY RATE) CIS-MQTL RESULTS. THE GENOME-WIDE SIGNIFICANT AND SUBTHRESHOLD (P < 10(-4)) GWAS SNPS WERE ENRICHED IN THE SIGNIFICANT MQTL SNPS (HYPERGEOMETRIC TEST P < 0.00001). WE OBSERVED ENRICHMENT FOR SITES LOCATED IN CPG SHORES AND SHELVES, BUT NOT CPG ISLANDS. USING BAYESIAN COLOCALIZATION, WE IDENTIFIED LOCI IN REGIONS NEAR KCNK3, EEFSEC, PIK3CD, DCDC2C, TCERG1L, FRMD4B, AND IL27. CONCLUSIONS: COLOCALIZATION OF MQTL AND GWAS LOCI PROVIDES REGULATORY CHARACTERIZATION OF SIGNIFICANT AND SUBTHRESHOLD GWAS FINDINGS, SUPPORTING A ROLE FOR GENETIC CONTROL OF METHYLATION IN COPD PATHOGENESIS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
5  5976 27 TET1-DEPENDENT EPIGENETIC MODIFICATION OF BDNF EXPRESSION IN DORSAL HORN NEURONS MEDIATES NEUROPATHIC PAIN IN RATS. TEN-ELEVEN TRANSLOCATION METHYLCYTOSINE DIOXYGENASE 1 (TET1) MEDIATES THE CONVERSION OF 5-METHYLCYTOSINE (5 MC) TO 5-HYDROXYMETHYLCYTOSINE (5 HMC), HENCE PROMOTING DNA DEMETHYLATION. ALTHOUGH RECENT STUDIES HAVE LINKED THE DNA DEMETHYLATION OF SPECIFIC GENES TO PAIN HYPERSENSITIVITY, THE ROLE OF SPINAL TET1-DEPENDENT DNA DEMETHYLATION IN NOCICEPTION HYPERSENSITIVITY DEVELOPMENT REMAINS ELUSIVE. HERE, WE REPORT CORRELATED WITH BEHAVIORAL ALLODYNIA, SPINAL NERVE LIGATION (SNL) UPREGULATED TET1 EXPRESSION IN DORSAL HORN NEURONS THAT HYDROXYLATE 5 MC TO 5 HMC AT CPG DINUCLEOTIDES IN THE BDNF PROMOTER TO PROMOTE SPINAL BDNF EXPRESSION AT DAY 7 AFTER OPERATION. FOCAL KNOCKDOWN OF SPINAL TET1 EXPRESSION DECREASED TET1 BINDING AND 5 HMC ENRICHMENT, FURTHER INCREASED 5 MC ENRICHMENT AT CPG SITES IN THE BDNF PROMOTER AND DECREASED SPINAL BDNF EXPRESSION ACCOMPANIED BY THE ALLEVIATION OF THE DEVELOPED ALLODYNIA. MOREOVER, AT DAY 7 AFTER OPERATION, SNL-ENHANCED TET1 EXPRESSION ALSO INHIBITED THE BINDING OF DNA METHYLTRANSFERASES (DNMTS, I.E., DNMT1, DNMT3A, AND DNMT3B) TO THE BDNF PROMOTER, A REQUIREMENT FOR TRANSCRIPTIONAL SILENCING BY CATALYSING 5-CYTOSINE (5C) TO 5 MC. TOGETHER, THESE DATA SUGGEST AT CPG SITES OF THE BDNF PROMOTER, SNL-ENHANCED TET1 EXPRESSION PROMOTES DNA DEMETHYLATION BOTH BY CONVERTING 5 MC TO 5 HMC AND INHIBITING DNMT BINDING TO REGULATE SPINAL BDNF EXPRESSION, HENCE CONTRIBUTING TO BEHAVIORAL ALLODYNIA DEVELOPMENT.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
6  2481 23 EPIGENETIC UPREGULATION OF LNCRNAS AT 13Q14.3 IN LEUKEMIA IS LINKED TO THE IN CIS DOWNREGULATION OF A GENE CLUSTER THAT TARGETS NF-KB. NON-CODING RNAS ARE MUCH MORE COMMON THAN PREVIOUSLY THOUGHT. HOWEVER, FOR THE VAST MAJORITY OF NON-CODING RNAS, THE CELLULAR FUNCTION REMAINS ENIGMATIC. THE TWO LONG NON-CODING RNA (LNCRNA) GENES DLEU1 AND DLEU2 MAP TO A CRITICAL REGION AT CHROMOSOMAL BAND 13Q14.3 THAT IS RECURRENTLY DELETED IN SOLID TUMORS AND HEMATOPOIETIC MALIGNANCIES LIKE CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). WHILE NO POINT MUTATIONS HAVE BEEN FOUND IN THE PROTEIN CODING CANDIDATE GENES AT 13Q14.3, THEY ARE DEREGULATED IN MALIGNANT CELLS, SUGGESTING AN EPIGENETIC TUMOR SUPPRESSOR MECHANISM. WE THEREFORE CHARACTERIZED THE EPIGENETIC MAKEUP OF 13Q14.3 IN CLL CELLS AND FOUND HISTONE MODIFICATIONS BY CHROMATIN-IMMUNOPRECIPITATION (CHIP) THAT ARE ASSOCIATED WITH ACTIVATED TRANSCRIPTION AND SIGNIFICANT DNA-DEMETHYLATION AT THE TRANSCRIPTIONAL START SITES OF DLEU1 AND DLEU2 USING 5 DIFFERENT SEMI-QUANTITATIVE AND QUANTITATIVE METHODS (APRIMES, BIOCOBRA, MCIP, MASSARRAY, AND BISULFITE SEQUENCING). THESE EPIGENETIC ABERRATIONS WERE CORRELATED WITH TRANSCRIPTIONAL DEREGULATION OF THE NEIGHBORING CANDIDATE TUMOR SUPPRESSOR GENES, SUGGESTING A COREGULATION IN CIS OF THIS GENE CLUSTER. WE FOUND THAT THE 13Q14.3 GENES IN ADDITION TO THEIR PREVIOUSLY KNOWN FUNCTIONS REGULATE NF-KB ACTIVITY, WHICH WE COULD SHOW AFTER OVEREXPRESSION, SIRNA-MEDIATED KNOCKDOWN, AND DOMINANT-NEGATIVE MUTANT GENES BY USING WESTERN BLOTS WITH PREVIOUSLY UNDESCRIBED ANTIBODIES, BY A CUSTOMIZED ELISA AS WELL AS BY REPORTER ASSAYS. IN ADDITION, WE PERFORMED AN UNBIASED SCREEN OF 810 HUMAN MIRNAS AND IDENTIFIED THE MIR-15/16 FAMILY OF GENES AT 13Q14.3 AS THE STRONGEST INDUCERS OF NF-KB ACTIVITY. IN SUMMARY, THE TUMOR SUPPRESSOR MECHANISM AT 13Q14.3 IS A CLUSTER OF GENES CONTROLLED BY TWO LNCRNA GENES THAT ARE REGULATED BY DNA-METHYLATION AND HISTONE MODIFICATIONS AND WHOSE MEMBERS ALL REGULATE NF-KB. THEREFORE, THE TUMOR SUPPRESSOR MECHANISM IN 13Q14.3 UNDERLINES THE ROLE BOTH OF EPIGENETIC ABERRATIONS AND OF LNCRNA GENES IN HUMAN TUMORIGENESIS AND IS AN EXAMPLE OF COLOCALIZATION OF A FUNCTIONALLY RELATED GENE CLUSTER.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
7  3332 40 HISTONE DEACETYLASE INHIBITOR-INDUCED EMERGENCE OF SYNAPTIC DELTA-OPIOID RECEPTORS AND BEHAVIORAL ANTINOCICEPTION IN PERSISTENT NEUROPATHIC PAIN. THE EFFICACY OF OPIOIDS IN PATIENTS WITH CHRONIC NEUROPATHIC PAIN REMAINS CONTROVERSIAL. ALTHOUGH ACTIVATION OF DELTA-OPIOID RECEPTORS (DORS) IN THE BRAINSTEM REDUCES INFLAMMATION-INDUCED PERSISTENT HYPERALGESIA, IT IS NOT EFFECTIVE UNDER PERSISTENT NEUROPATHIC PAIN CONDITIONS AND THESE CLINICAL PROBLEMS REMAIN LARGELY UNKNOWN. IN THIS STUDY, BY USING A CHRONIC CONSTRICTION INJURY (CCI) OF THE SCIATIC NERVE IN RATS, WE FOUND THAT IN THE BRAINSTEM NUCLEUS RAPHE MAGNUS (NRM), DORS EMERGED ON THE SURFACE MEMBRANE OF CENTRAL SYNAPTIC TERMINALS ON DAY 3 AFTER CCI SURGERY AND DISAPPEARED ON DAY 14. HISTONE DEACETYLASE (HDAC) INHIBITORS MICROINJECTED INTO THE NRM IN VIVO INCREASED THE LEVEL OF SYNAPTOSOMAL DOR PROTEIN AND NRM INFUSION OF DOR AGONISTS PRODUCING AN ANTINOCICEPTIVE EFFECT IN A NERVE GROWTH FACTOR (NGF) SIGNALING-DEPENDENT MANNER. IN VITRO, IN CCI RAT SLICES INCUBATED WITH HDAC INHIBITORS, DOR AGONISTS SIGNIFICANTLY INHIBITED EPSCS. THIS EFFECT WAS BLOCKED BY TYROSINE RECEPTOR KINASE A ANTAGONISTS. CHROMATIN IMMUNOPRECIPITATION ANALYSIS REVEALED THAT NRM INFUSION OF HDAC INHIBITORS IN CCI RATS INCREASED THE LEVEL OF HISTONE H4 ACETYLATION AT NGF GENE PROMOTER REGIONS. NGF WAS INFUSED INTO THE NRM OR INCUBATED CCI RAT SLICES DROVE DORS TO THE SURFACE MEMBRANE OF SYNAPTIC TERMINALS. TAKEN TOGETHER, EPIGENETIC UPREGULATION OF NGF ACTIVITY BY HDAC INHIBITORS IN THE NRM PROMOTES THE TRAFFICKING OF DORS TO PAIN-MODULATING NEURONAL SYNAPSES UNDER NEUROPATHIC PAIN CONDITIONS, LEADING TO DELTA-OPIOID ANALGESIA. THESE FINDINGS INDICATE THAT THERAPEUTIC USE OF DOR AGONISTS COMBINED WITH HDAC INHIBITORS MIGHT BE EFFECTIVE IN CHRONIC NEUROPATHIC PAIN MANAGEMENTS.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
8  2300 37 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
9  3809 35 INTRAPERITONEAL 5-AZACYTIDINE ALLEVIATES NERVE INJURY-INDUCED PAIN IN RATS BY MODULATING DNA METHYLATION. TO INVESTIGATE THE ROLE OF DNA METHYLATION IN MODULATING CHRONIC NEUROPATHIC PAIN (NPP), IDENTIFY POSSIBLE TARGET GENES OF DNA METHYLATION INVOLVED IN THIS PROCESS, AND PRELIMINARILY CONFIRM THE MEDICINAL VALUE OF THE DNA METHYLTRANSFERASES (DNMTS) INHIBITOR 5-AZACYTIDINE (5-AZA) IN NPP BY TARGETING GENE METHYLATION. TWO RAT NPP MODELS, CHRONIC CONSTRICTION INJURY (CCI) AND SPINAL NERVE LIGATION (SNL), WERE USED. THE DNA METHYLATION PROFILES IN THE LUMBAR SPINAL CORD WERE ASSAYED USING AN ARRAYSTAR RAT REFSEQ PROMOTER ARRAY. THE UNDERLYING GENES WITH DIFFERENTIAL METHYLATION WERE THEN IDENTIFIED AND SUBMITTED TO GENE ONTOLOGY AND PATHWAY ANALYSIS. METHYL-DNA IMMUNOPRECIPITATION QUANTITATIVE PCR (MEDIP-QPCR) AND QUANTITATIVE REVERSE TRANSCRIPTION-PCR (RT-QPCR) WERE USED TO CONFIRM GENE METHYLATION AND EXPRESSION. THE PROTECTIVE FUNCTION OF 5-AZA IN NPP AND GENE EXPRESSION WERE EVALUATED VIA BEHAVIORAL ASSAYS AND RT-QPCR, RESPECTIVELY. ANALYSIS OF THE DNA METHYLATION PATTERNS IN THE LUMBAR SPINAL CORD INDICATED THAT 1205 DIFFERENTIALLY METHYLATED FRAGMENTS IN CCI RATS WERE LOCATED WITHIN DNA PROMOTER REGIONS, INCLUDING 638 HYPERMETHYLATED FRAGMENTS AND 567 HYPOMETHYLATED FRAGMENTS. THE METHYLATION LEVELS OF GRM4, HTR4, ADRB2, KCNF1, GAD2, AND PPARG, WHICH ARE ASSOCIATED WITH LONG-TERM POTENTIATION (LTP) AND GLUTAMATERGIC SYNAPSE PATHWAYS, WERE INCREASED WITH A CORRESPONDING DECREASE IN THEIR MRNA EXPRESSION, IN THE SPINAL CORDS OF CCI RATS. MOREOVER, WE FOUND THAT THE INTRAPERITONEAL INJECTION OF 5-AZA (4 MG/KG) ATTENUATED CCI- OR SNL-INDUCED MECHANICAL ALLODYNIA AND THERMAL HYPERALGESIA. FINALLY, THE MRNA EXPRESSION OF HYPERMETHYLATED GENES SUCH AS GRM4, HTR4, ADRB2, KCNF1, AND GAD2 WAS REVERSED AFTER 5-AZA TREATMENT. CCI INDUCED WIDESPREAD METHYLATION CHANGES IN THE DNA PROMOTER REGIONS IN THE LUMBAR SPINAL CORD. INTRAPERITONEAL 5-AZA ALLEVIATED HYPERALGESIA IN CCI AND SNL RATS, AN EFFECT ACCOMPANIED BY THE REVERSED EXPRESSION OF HYPERMETHYLATED GENES. THUS, DNA METHYLATION INHIBITION REPRESENTS A PROMISING EPIGENETIC STRATEGY FOR PROTECTION AGAINST CHRONIC NPP FOLLOWING NERVE INJURY. OUR STUDY LAYS A THEORETICAL FOUNDATION FOR 5-AZA TO BECOME A CLINICAL TARGETED DRUG.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
10  3459 23 HYPOMETHYLATION OF NERVE GROWTH FACTOR (NGF) PROMOTES BINDING OF C/EBPALPHA AND CONTRIBUTES TO INFLAMMATORY HYPERALGESIA IN RATS. BACKGROUND: CHRONIC PAIN USUALLY ACCOMPANIED BY TISSUE DAMAGE AND INFLAMMATION. HOWEVER, THE PATHOGENESIS OF CHRONIC PAIN REMAINS UNCLEAR. METHODS: WE INVESTIGATED THE ROLE OF NERVE GROWTH FACTOR (NGF) IN CHRONIC INFLAMMATORY PAIN INDUCED BY COMPLETE FREUND'S ADJUVANT (CFA), EXPLORED THE METHYLATION STATUS OF CPG ISLANDS IN THE PROMOTER REGION OF THE NGF GENE, AND CLARIFIED THE FUNCTION AND MECHANISM OF C/EBPALPHA-NGF SIGNALING PATHWAY FROM EPIGENETIC PERSPECTIVE IN THE CHRONIC INFLAMMATORY PAIN MODEL. RESULTS: CFA INDUCED SIGNIFICANT HYPERALGESIA AND CONTINUOUS UPREGULATION OF NGF MRNA AND PROTEIN LEVELS IN THE L4-6 DORSAL ROOT GANGLIONS (DRGS) IN RATS. HYPOMETHYLATION OF CPG ISLANDS OCCURRED IN THE NGF GENE PROMOTER REGION AFTER CFA TREATMENT. AT THE SAME TIME, THE MIR-29B EXPRESSION LEVEL WAS SIGNIFICANTLY INCREASED, WHILE THE DNA METHYLTRANSFERASE 3B (DNMT3B) LEVEL REDUCED SIGNIFICANTLY. MOREOVER, CFA TREATMENT PROMOTED BINDING OF C/EBPALPHA TO THE NGF GENE PROMOTER REGION AND C/EBPALPHA SIRNA TREATMENT OBVIOUSLY DECREASED EXPRESSION OF NGF LEVELS AND ALSO ALLEVIATE INFLAMMATORY HYPERALGESIA SIGNIFICANTLY IN RATS. CONCLUSION: COLLECTIVELY, THE RESULTS INDICATED THAT CFA LEADS TO THE UPREGULATION OF MIR-29B LEVEL, WHICH REPRESSES THE EXPRESSION OF DNMT3B, ENHANCES THE DEMETHYLATION OF THE NGF GENE PROMOTER REGION, AND PROMOTES THE BINDING OF C/EBPALPHA WITH THE NGF GENE PROMOTER, THUS RESULTS IN THE UPREGULATION OF NGF GENE EXPRESSION AND MAINTENANCE OF CHRONIC INFLAMMATORY PAIN.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
11  6660 29 UPREGULATION OF CXCR4 THROUGH PROMOTER DEMETHYLATION CONTRIBUTES TO INFLAMMATORY HYPERALGESIA IN RATS. AIM AND METHODS: CHRONIC PAIN ASSOCIATED WITH INFLAMMATION IS A COMMON CLINICAL PROBLEM, AND THE UNDERLYING MECHANISMS YET ARE INCOMPLETELY DEFINED. DNA METHYLATION HAS BEEN IMPLICATED IN THE PATHOGENESIS OF CHRONIC PAIN. HOWEVER, THE SPECIFIC GENES REGULATED BY DNA METHYLATION UNDER INFLAMMATORY PAIN CONDITION REMAIN LARGELY UNKNOWN. HERE, WE INVESTIGATED HOW CHEMOKINE RECEPTOR CXCR4 EXPRESSION IS REGULATED BY DNA METHYLATION AND HOW IT CONTRIBUTES TO INFLAMMATORY PAIN INDUCED BY COMPLETE FREUND'S ADJUVANT (CFA) IN RATS. RESULTS: INTRAPLANTAR INJECTION OF CFA COULD NOT ONLY INDUCE SIGNIFICANT HYPERALGESIA IN RATS, BUT ALSO SIGNIFICANTLY INCREASE THE EXPRESSION OF CXCR4 MRNA AND PROTEIN IN THE DORSAL ROOT GANGLION (DRG). INTRATHECAL INJECTION OF CXCR4 ANTAGONIST AMD3100 SIGNIFICANTLY RELIEVED HYPERALGESIA IN INFLAMMATORY RATS IN A TIME- AND DOSE-DEPENDENT MANNER. BISULFITE SEQUENCING AND METHYLATION-SPECIFIC PCR DEMONSTRATE THAT CFA INJECTION LED TO A SIGNIFICANT DEMETHYLATION OF CPG ISLAND AT CXCR4 GENE PROMOTER. CONSISTENTLY, THE EXPRESSION OF DNMT3B WAS SIGNIFICANTLY DOWNREGULATED AFTER CFA INJECTION. ONLINE SOFTWARE PREDICTION REVEALS THREE BINDING SITES OF P65 IN THE CPG ISLAND OF CXCR4 GENE PROMOTER, WHICH HAS CONFIRMED BY THE CHROMATIN IMMUNOPRECIPITATION ASSAY, CFA TREATMENT SIGNIFICANTLY INCREASES THE RECRUITMENT OF P65 TO CXCR4 GENE PROMOTER. INHIBITION OF NF-KB SIGNALING USING P65 INHIBITOR PYRROLIDINE DITHIOCARBAMATE SIGNIFICANTLY PREVENTED THE INCREASES OF THE CXCR4 EXPRESSION. CONCLUSION: UPREGULATION OF CXCR4 EXPRESSION DUE TO PROMOTER DEMETHYLATION FOLLOWED BY INCREASED RECRUITMENT OF P65 TO PROMOTER OF CXCR4 GENE CONTRIBUTES TO INFLAMMATORY HYPERALGESIA. THESE FINDINGS PROVIDE A THEORETICAL BASIS FOR THE TREATMENT OF CHRONIC PAIN FROM AN EPIGENETIC PERSPECTIVE.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
12  2479 30 EPIGENETIC UPREGULATION OF CXCL12 EXPRESSION MEDIATES ANTITUBULIN CHEMOTHERAPEUTICS-INDUCED NEUROPATHIC PAIN. CLINICALLY, MICROTUBULE-TARGETED AGENTS-INDUCED NEUROPATHIC PAIN HAMPERS CHEMOTHERAPEUTICS FOR PATIENTS WITH CANCER. HERE, WE FOUND THAT APPLICATION OF PACLITAXEL OR VINCRISTINE INCREASED THE PROTEIN AND MRNA EXPRESSION OF CXCL12 AND FREQUENCY AND AMPLITUDE OF MINIATURE EXCITATORY POST SYNAPTIC CURRENTS (MEPSCS) IN SPINAL DORSAL HORN NEURONS. SPINAL LOCAL APPLICATION OF CXCL12 INDUCED THE LONG-TERM POTENTIATION OF NOCICEPTIVE SYNAPTIC TRANSMISSION AND INCREASED THE AMPLITUDE OF MEPSCS. INHIBITION OF CXCL12 USING THE TRANSGENIC MICE (CXCL12) OR NEUTRALIZING ANTIBODY OR SIRNA AMELIORATED THE MEPSC'S ENHANCEMENT AND MECHANICAL ALLODYNIA. IN ADDITION, PACLITAXEL AND VINCRISTINE BOTH COULD INCREASE THE PHOSPHORYLATION OF SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION 3 (STAT3) AND THE ACETYLATION OF HISTONE H4 IN THE CXCL12-EXPRESSING NEURONS. IMMUNOPRECIPITATION AND CHROMATIN IMMUNOPRECIPITATION ASSAYS DEMONSTRATED THAT ANTITUBULIN CHEMOTHERAPEUTICS INCREASED THE BINDING OF STAT3 TO THE CXCL12 GENE PROMOTER AND THE INTERACTION BETWEEN STAT3 AND P300, AND CONTRIBUTED TO THE ENHANCED TRANSCRIPTION OF CXCL12 BY INCREASING THE ACETYLATION OF HISTONE H4 IN CXCL12 GENE PROMOTER. INHIBITION OF STAT3 BY INTRATHECAL INJECTION OF ADENO-ASSOCIATED VIRUS ENCODING CRE AND GREEN FLUORESCENT PROTEIN INTO STAT3 MICE OR INHIBITOR S3I-201 INTO RATS SUPPRESSED THE CXCL12 UPSURGE BY DECREASING THE ACETYLATION OF HISTONE H4. FINALLY, BLOCKADE OF CXCR4 BUT NOT CXCR7 AMELIORATED THE PACLITAXEL- OR VINCRISTINE-INDUCED MECHANICAL ALLODYNIA. TOGETHER, THESE RESULTS SUGGESTED THAT ENHANCED INTERACTION BETWEEN STAT3 AND P300 MEDIATED THE EPIGENETIC UPREGULATION OF CXCL12 IN DORSAL HORN NEURONS, WHICH CONTRIBUTED TO THE ANTITUBULIN CHEMOTHERAPEUTICS-INDUCED PERSISTENT PAIN.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
13  5266 34 PROMOTED INTERACTION OF C/EBPALPHA WITH DEMETHYLATED CXCR3 GENE PROMOTER CONTRIBUTES TO NEUROPATHIC PAIN IN MICE. DNA METHYLATION HAS BEEN IMPLICATED IN THE PATHOGENESIS OF CHRONIC PAIN. HOWEVER, THE SPECIFIC GENES REGULATED BY DNA METHYLATION UNDER NEUROPATHIC PAIN CONDITION REMAIN LARGELY UNKNOWN. HERE WE INVESTIGATED HOW CHEMOKINE RECEPTOR CXCR3 IS REGULATED BY DNA METHYLATION AND HOW IT CONTRIBUTES TO NEUROPATHIC PAIN INDUCED BY SPINAL NERVE LIGATION (SNL) IN MICE. SNL INCREASED CXCR3 MRNA AND PROTEIN EXPRESSION IN THE NEURONS OF THE SPINAL CORD. MEANWHILE, THE CPG (5'-CYTOSINE-PHOSPHATE-GUANINE-3') ISLAND IN THE CXCR3 GENE PROMOTER REGION WAS DEMETHYLATED, AND THE EXPRESSION OF DNA METHYLTRANSFERASE 3B (DNMT3B) WAS DECREASED. SNL ALSO INCREASED THE BINDING OF CCAAT (CYTIDINE-CYTIDINE-ADENOSINE-ADENOSINE-THYMIDINE)/ENHANCER BINDING PROTEIN ALPHA (C/EBPALPHA) WITH CXCR3 PROMOTER AND DECREASED THE BINDING OF DNMT3B WITH CXCR3 PROMOTER IN THE SPINAL CORD. C/EBPALPHA EXPRESSION WAS INCREASED IN SPINAL NEURONS AFTER SNL, AND INHIBITION OF C/EBPALPHA BY INTRATHECAL SMALL INTERFERING RNA ATTENUATED SNL-INDUCED PAIN HYPERSENSITIVITY AND REDUCED CXCR3 EXPRESSION. FURTHERMORE, SNL-INDUCED MECHANICAL ALLODYNIA AND HEAT HYPERALGESIA WERE MARKEDLY REDUCED IN CXCR3(-/-) MICE. SPINAL INHIBITION OF CXCR3 BY SHRNA OR CXCR3 ANTAGONIST ALSO ATTENUATED ESTABLISHED NEUROPATHIC PAIN. MOREOVER, CXCL10, THE LIGAND OF CXCR3, WAS INCREASED IN SPINAL NEURONS AND ASTROCYTES AFTER SNL. SUPERFUSING SPINAL CORD SLICES WITH CXCL10 ENHANCED SPONTANEOUS EPSCS AND POTENTIATED NMDA-INDUCED AND AMPA-INDUCED CURRENTS OF LAMINA II NEURONS. FINALLY, INTRATHECAL INJECTION OF CXCL10 INDUCED CXCR3-DEPENDENT PAIN HYPERSENSITIVITY IN NAIVE MICE. COLLECTIVELY, OUR RESULTS DEMONSTRATED THAT CXCR3, INCREASED BY DNA DEMETHYLATION AND THE ENHANCED INTERACTION WITH C/EBPALPHA, CAN BE ACTIVATED BY CXCL10 TO FACILITATE EXCITATORY SYNAPTIC TRANSMISSION AND CONTRIBUTE TO THE MAINTENANCE OF NEUROPATHIC PAIN. SIGNIFICANCE STATEMENT: PERIPHERAL NERVE INJURY INDUCES CHANGES OF GENE EXPRESSION IN THE SPINAL CORD THAT MAY CONTRIBUTE TO THE PATHOGENESIS OF NEUROPATHIC PAIN. CXCR3 IS A CHEMOKINE RECEPTOR. WHETHER IT IS INVOLVED IN NEUROPATHIC PAIN AND HOW IT IS REGULATED AFTER NERVE INJURY REMAIN LARGELY UNKNOWN. OUR STUDY DEMONSTRATES THAT SPINAL NERVE LIGATION DOWNREGULATES THE EXPRESSION OF DNMT3B, WHICH MAY CAUSE DEMETHYLATION OF CXCR3 GENE PROMOTER AND FACILITATE THE BINDING OF CCAAT/ENHANCER BINDING PROTEIN ALPHA WITH CXCR3 PROMOTER AND FURTHER INCREASE CXCR3 EXPRESSION IN SPINAL NEURONS. THE UPREGULATED CXCR3 MAY CONTRIBUTE TO NEUROPATHIC PAIN BY FACILITATING CENTRAL SENSITIZATION. OUR STUDY REVEALS AN EPIGENETIC MECHANISM UNDERLYING CXCR3 EXPRESSION AND ALSO SUGGESTS THAT TARGETING THE EXPRESSION OR ACTIVATION OF CXCR3 SIGNALING MAY OFFER NEW THERAPEUTICS FOR NEUROPATHIC PAIN.	2017	

14  4579 25 N(6)-METHYLADENOSINE METHYLASE METTL3 CONTRIBUTES TO NEUROPATHIC PAIN BY EPIGENETIC SILENCING OF MU OPIOID RECEPTOR. WE AIMED AT EXPLORING THE ROLE AND MECHANISM OF METTL3-MEDIATED M(6)A MODIFICATION IN NEUROPATHIC PAIN. MALE SPRAGUE-DAWLEY RATS WERE RANDOMLY DIVIDED INTO FOUR GROUPS: SHAM OPERATION GROUP (SHAM GROUP), CHRONIC CONSTRICTION INJURY (CCI) OF THE SCIATIC NERVE MODEL GROUP (NPP GROUP), INTRATHECAL INJECTION OF VIRUS DOWN-REGULATED METTL3 + CCI MODEL GROUP (M3 + NPP GROUP) AND INTRATHECAL INJECTION OF NEGATIVE CONTROL VIRUS + CCI MODEL GROUP (SCR + NPP GROUP). THE M3 + NPP GROUP AND THE SCR + NPP GROUP WERE INTRATHECALLY INJECTED WITH VIRUS NINETEEN DAYS BEFORE OPERATION. THE PAW WITHDRAWAL MECHANICAL THRESHOLDS AND PAW WITHDRAWAL LATENCY WERE RESPECTIVELY RECORDED ONE DAY BEFORE OPERATION, THREE DAYS, FIVE DAYS AND SEVEN DAYS AFTER OPERATION. THE RATS WERE SACRIFICED ON THE SEVENTH DAY AFTER OPERATION, AND THEIR SPINAL CORD TISSUES WERE TAKEN. THE FROZEN SECTIONS OF RATS WERE PERFORMED TO OBSERVE THE EXPRESSION OF GREEN FLUORESCENT PROTEIN OF THE VIRUS. THE METHYLATION LEVEL OF RNA, THE PROTEIN EXPRESSION OF M(6)A-RELATED ENZYME (METTL3) AND MU OPIOID RECEPTOR (MOR) IN SPINAL CORD TISSUES OF THE FOUR GROUPS WERE MEASURED. DOWNREGULATION OF METTL3 HAD NO EFFECT ON THE OVERALL METHYLATION LEVEL OF THE SPINAL CORD, BUT IT COULD REGULATE THE METHYLATION LEVEL OF THE OPRM1 GENE RNA ENCODING MOR, PARTIALLY RESTORE THE EXPRESSION OF MOR, AND RELIEVE PAIN IN RATS. IN THE PROCESS OF NPP, METTL3 MAY INHIBIT THE EXPRESSION OF MOR BY REGULATING THE METHYLATION LEVEL OF OPRM1 GENE RNA ENCODING MOR, AND ULTIMATELY PROMOTE THE OCCURRENCE AND DEVELOPMENT OF NPP.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
15   717 37 CALCITONIN GENE-RELATED PEPTIDE REGULATES SPINAL MICROGLIAL ACTIVATION THROUGH THE HISTONE H3 LYSINE 27 TRIMETHYLATION VIA ENHANCER OF ZESTE HOMOLOG-2 IN RATS WITH NEUROPATHIC PAIN. BACKGROUND: CALCITONIN GENE-RELATED PEPTIDE (CGRP) AS A MEDIATOR OF MICROGLIAL ACTIVATION AT THE TRANSCRIPTIONAL LEVEL MAY FACILITATE NOCICEPTIVE SIGNALING. TRIMETHYLATION OF H3 LYSINE 27 (H3K27ME3) BY ENHANCER OF ZESTE HOMOLOG 2 (EZH2) IS AN EPIGENETIC MARK THAT REGULATES INFLAMMATORY-RELATED GENE EXPRESSION AFTER PERIPHERAL NERVE INJURY. IN THIS STUDY, WE EXPLORED THE RELATIONSHIP BETWEEN CGRP AND H3K27ME3 IN MICROGLIAL ACTIVATION AFTER NERVE INJURY, AND ELUCIDATED THE UNDERLYING MECHANISMS IN THE PATHOGENESIS OF CHRONIC NEUROPATHIC PAIN. METHODS: MICROGLIAL CELLS (BV2) WERE TREATED WITH CGRP AND DIFFERENTIALLY ENRICHMENTS OF H3K27ME3 ON GENE PROMOTERS WERE EXAMINED USING CHIP-SEQ. A CHRONIC CONSTRICTION INJURY (CCI) RAT MODEL WAS USED TO EVALUATE THE ROLE OF CGRP ON MICROGLIAL ACTIVATION AND EZH2/H3K27ME3 SIGNALING IN CCI-INDUCED NEUROPATHIC PAIN. RESULTS: OVEREXPRESSIONS OF EZH2 AND H3K27ME3 WERE CONFIRMED IN SPINAL MICROGLIA OF CCI RATS BY IMMUNOFLUORESCENCE. CGRP TREATMENT INDUCED THE INCREASED OF H3K27ME3 EXPRESSION IN THE SPINAL DORSAL HORN AND CULTURED MICROGLIAL CELLS (BV2) THROUGH EZH2. CHIP-SEQ DATA INDICATED THAT CGRP SIGNIFICANTLY ALTERED H3K27ME3 ENRICHMENTS ON GENE PROMOTERS IN MICROGLIA FOLLOWING CGRP TREATMENT, INCLUDING 173 GAINING H3K27ME3 AND 75 LOSING THIS MARK, WHICH MOSTLY ENRICHED IN REGULATION OF CELL GROWTH, PHAGOSOME, AND INFLAMMATION. QRT-PCR VERIFIED EXPRESSIONS OF REPRESENTATIVE CANDIDATE GENES (TRAF3IP2, BCL2L11, ITGAM, DAB2, NLRP12, WNT3, ADAM10) AND REAL-TIME CELL ANALYSIS (RTCA) VERIFIED MICROGLIAL PROLIFERATION. ADDITIONALLY, CGRP TREATMENT AND CCI INCREASED EXPRESSIONS OF ITGAM, ADAM10, MCP-1, AND CX3CR1, KEY MEDIATORS OF MICROGLIAL ACTIVATION IN SPINAL DORSAL HORN AND CULTURED MICROGLIAL CELLS. SUCH INCREASED EFFECTS INDUCED BY CCI WERE SUPPRESSED BY CGRP ANTAGONIST AND EZH2 INHIBITOR, WHICH WERE CONCURRENTLY ASSOCIATED WITH THE ATTENUATED MECHANICAL AND THERMAL HYPERALGESIA IN CCI RATS. CONCLUSION: OUR FINDINGS HIGHLY INDICATE THAT CGRP IS IMPLICATED IN THE GENESIS OF NEUROPATHIC PAIN THROUGH REGULATING MICROGLIAL ACTIVATION VIA EZH2-MEDIATED H3K27ME3 IN THE SPINAL DORSAL HORN.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
16  4366 25 MIRNA-23A/CXCR4 REGULATES NEUROPATHIC PAIN VIA DIRECTLY TARGETING TXNIP/NLRP3 INFLAMMASOME AXIS. BACKGROUND: CHEMOKINE CXC RECEPTOR 4 (CXCR4) IN SPINAL GLIAL CELLS HAS BEEN IMPLICATED IN NEUROPATHIC PAIN. HOWEVER, THE REGULATORY CASCADES OF CXCR4 IN NEUROPATHIC PAIN REMAIN ELUSIVE. HERE, WE INVESTIGATED THE FUNCTIONAL REGULATORY ROLE OF MIRNAS IN THE PAIN PROCESS AND ITS INTERPLAY WITH CXCR4 AND ITS DOWNSTREAM SIGNALING. METHODS: MIRNAS AND CXCR4 AND ITS DOWNSTREAM SIGNALING MOLECULES WERE MEASURED IN THE SPINAL CORDS OF MICE WITH SCIATIC NERVE INJURY VIA PARTIAL SCIATIC NERVE LIGATION (PSNL). IMMUNOBLOTTING, IMMUNOFLUORESCENCE, IMMUNOPRECIPITATION, AND MAMMAL TWO-HYBRID AND BEHAVIORAL TESTS WERE USED TO EXPLORE THE DOWNSTREAM CXCR4-DEPENDENT SIGNALING PATHWAY. RESULTS: CXCR4 EXPRESSION INCREASED IN SPINAL GLIAL CELLS OF MICE WITH PSNL-INDUCED NEUROPATHIC PAIN. BLOCKING CXCR4 ALLEVIATED THE PAIN BEHAVIOR; CONTRARILY, OVEREXPRESSING CXCR4 INDUCED PAIN HYPERSENSITIVITY. MICRORNA-23A-3P (MIR-23A) DIRECTLY BOUNDS TO 3' UTR OF CXCR4 MRNA. PSNL-INDUCED NEUROPATHIC PAIN SIGNIFICANTLY REDUCED MRNA EXPRESSION OF MIR-23A. OVEREXPRESSION OF MIR-23A BY INTRATHECAL INJECTION OF MIR-23A MIMICS OR LENTIVIRUS REDUCED SPINAL CXCR4 AND PREVENTED PSNL-INDUCED NEUROPATHIC PAIN. IN CONTRAST, KNOCKDOWN OF MIR-23A BY INTRATHECAL INJECTION OF MIR-23A INHIBITOR OR LENTIVIRUS INDUCED PAIN-LIKE BEHAVIOR, WHICH WAS REDUCED BY CXCR4 INHIBITION. ADDITIONALLY, MIR-23A KNOCKDOWN OR CXCR4 OVEREXPRESSION IN NAIVE MICE COULD INCREASE THE THIOREDOXIN-INTERACTING PROTEIN (TXNIP), WHICH WAS ASSOCIATED WITH INDUCTION OF NOD-LIKE RECEPTOR PROTEIN 3 (NLRP3) INFLAMMASOME. INDEED, CXCR4 AND TXNIP WERE CO-EXPRESSED. THE MAMMAL TWO-HYBRID ASSAY REVEALED THE DIRECT INTERACTION BETWEEN CXCR4 AND TXNIP, WHICH WAS INCREASED IN THE SPINAL CORD OF PSNL MICE. IN PARTICULAR, INHIBITION OF TXNIP REVERSED PAIN BEHAVIOR ELICITED BY PSNL, MIR-23A KNOCKDOWN, OR CXCR4 OVEREXPRESSION. MOREOVER, MIR-23A OVEREXPRESSION OR CXCR4 KNOCKDOWN INHIBITED THE INCREASE OF TXNIP AND NLRP3 INFLAMMASOME IN PSNL MICE. CONCLUSIONS: MIR-23A, BY DIRECTLY TARGETING CXCR4, REGULATES NEUROPATHIC PAIN VIA TXNIP/NLRP3 INFLAMMASOME AXIS IN SPINAL GLIAL CELLS. EPIGENETIC INTERVENTIONS AGAINST MIR-23A, CXCR4, OR TXNIP MAY POTENTIALLY SERVE AS NOVEL THERAPEUTIC AVENUES IN TREATING PERIPHERAL NERVE INJURY-INDUCED NOCICEPTIVE HYPERSENSITIVITY.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
17  1631 32 DNMT3A METHYLATION IN NEUROPATHIC PAIN. BACKGROUND: MU OPIOID RECEPTOR (MOR) PLAYS A CRUCIAL ROLE IN MEDIATING ANALGESIC EFFECTS OF OPIOIDS AND IS CLOSELY ASSOCIATED WITH THE PATHOLOGIES OF NEUROPATHIC PAIN. PREVIOUS STUDIES HAVE REPORTED THAT PERIPHERAL NERVE INJURY DOWNREGULATES MOR EXPRESSION, BUT THE EPIGENETIC MECHANISMS REMAIN UNKNOWN. OBJECTIVE: THEREFORE, WE INVESTIGATED DNA METHYLTRANSFERASE3A (DNMT3A) EXPRESSION OR METHYLATION CHANGES WITHIN MOR PROMOTER IN THE SPINAL CORD IN A NEUROPATHIC PAIN INDUCED BY A CHRONIC CONSTRICTION INJURY (CCI) MOUSE MODEL AND FURTHER DETERMINED WHETHER THESE INJURY-ASSOCIATED CHANGES ARE REVERSIBLE BY PHARMACOLOGICAL INTERVENTIONS. METHODS: A CCI MOUSE MODEL WAS ESTABLISHED AND TISSUE SPECIMENS OF LUMBAR SPINAL CORDS WERE COLLECTED. THE NOCICEPTION THRESHOLD WAS EVALUATED BY A MODEL HEATED 400 BASE. DNMT3A AND MOR MRNA AND PROTEIN LEVEL WERE DETECTED BY REAL-TIME-POLYMERASE CHAIN REACTION AND WESTERN BLOT, RESPECTIVELY. METHYLATION OF DNMT3A GENE WAS MEASURED BY METHYLATION-SPECIFIC PCR. RESULTS: OUR DATA SHOWED THAT CHRONIC NERVE INJURY LED TO A SIGNIFICANT UPREGULATION OF DNMT3A EXPRESSION THAT WAS ASSOCIATED WITH INCREASED METHYLATION OF MOR GENE PROMOTER AND DECREASED MOR PROTEIN EXPRESSION IN THE SPINAL CORD. INHIBITION OF DNMT3A CATALYTIC ACTIVITY WITH DNMT INHIBITOR RG108 SIGNIFICANTLY BLOCKED THE INCREASE IN METHYLATION OF THE MOR PROMOTER, AND THEN UPREGULATED MOR EXPRESSION AND ATTENUATED THERMAL HYPERALGESIA IN NEUROPATHIC PAIN MICE. CONCLUSION: THIS STUDY DEMONSTRATES THAT AN INCREASE OF DNMT3A EXPRESSION AND MOR METHYLATION EPIGENETICALLY PLAY AN IMPORTANT ROLE IN NEUROPATHIC PAIN. TARGETING DNMT3A TO THE PROMOTER OF MOR GENE BY DNMT INHIBITOR MAY BE A PROMISING APPROACH TO THE DEVELOPMENT OF NEW NEUROPATHIC PAIN THERAPY.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
18  2326 24 EPIGENETIC REGULATION OF HOTAIR IN ADVANCED CHRONIC MYELOID LEUKEMIA. PURPOSE: CHRONIC MYELOID LEUKEMIA (CML) ACCOUNTS FOR ~10% OF LEUKEMIA CASES, AND ITS PROGRESSION INVOLVES EPIGENETIC GENE REGULATION. THIS STUDY INVESTIGATED EPIGENETIC REGULATION OF HOTAIR AND ITS TARGET MICRORNA, MIR-143, IN ADVANCED CML. PATIENTS AND METHODS: WE FIRST ISOLATED BONE MARROW MONONUCLEAR CELLS FROM 70 PATIENTS WITH DIFFERENT PHASES OF CML AND FROM HEALTHY DONORS AS NORMAL CONTROL; WE ALSO CULTURED K562 AND KCL22 CELLS, TREATED WITH DEMETHYLATION DRUG; MTT ASSAY, FLOW CYTOMETRY, QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QPCR), METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP), WESTERN BLOT, LUCIFERASE ASSAY, RNA PULL-DOWN ASSAY AND RNA-BINDING PROTEIN IMMUNOPRECIPITATION (RIP) ASSAY WERE PERFORMED. RESULT: AS MEASURED BY QPCR, HOTAIR EXPRESSION IN K562 CELLS, KCL22 CELLS, AND SAMPLES FROM CASES OF ADVANCED-STAGE CML INCREASED WITH LEVELS OF SEVERAL DNA METHYLTRANSFERASES AND HISTONE DEACETYLATES, INCLUDING DNMT1, DNMT3A, HDAC1, EZH2, AND LSD1, AND MIR-143 LEVELS WERE DECREASED AND HOTAIR LEVELS WERE INCREASED. TREATMENT WITH 5-AZACYTIDINE, A DNA METHYLATION INHIBITOR, DECREASED DNMT1, DNMT3A, HDAC1, EZH2, LSD1 MRNA, PROTEIN LEVELS, AND HOTAIR MRNA LEVELS BUT INCREASED MIR-143 LEVELS. HOTAIR KNOCKDOWN AND MIR-143 OVEREXPRESSION BOTH INHIBITED PROLIFERATION AND PROMOTED APOPTOSIS IN KCL22 AND K562 CELLS THROUGH THE PI3K/AKT PATHWAY. RNA PULL-DOWN, MASS SPECTROMETRY, AND RIP ASSAYS SHOWED THAT HOTAIR INTERACTED WITH EZH2 AND LSD1. A DUAL-LUCIFERASE ASSAY DEMONSTRATED THAT HOTAIR INTERACTED WITH MIR-143. CONCLUSION: OUR FINDINGS DEMONSTRATE THE KEY EPIGENETIC INTERACTIONS OF HOTAIR RELATED TO CML PROGRESSION AND SUGGEST HOTAIR AS A POTENTIAL THERAPEUTIC TARGET FOR ADVANCED CML. FURTHERMORE, OUR RESULTS SUPPORT THE USE OF DEMETHYLATION DRUGS AS A CML TREATMENT STRATEGY.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
19  5459 23 RESEARCH ON THE EPIGENETIC REGULATION MECHANISM OF THE PTPN6 GENE IN ADVANCED CHRONIC MYELOID LEUKAEMIA. PTPN6, A TYROSINE PHOSPHATASE PROTEIN, PLAYS A NEGATIVE ROLE IN CELL SIGNAL TRANSDUCTION AND IS NEGATIVELY CORRELATED WITH TUMOUR FORMATION AND GROWTH. HOWEVER, EPIGENETIC REGULATION MECHANISM OF THE PTPN6 GENE IN ADVANCED CHRONIC MYELOID LEUKAEMIA (CML) REMAINS UNCLEAR. THIS STUDY INVESTIGATED BONE MARROW OR BLOOD SAMPLES FROM 44 CML PATIENTS AND 10 HEALTHY VOLUNTEERS. KCL22 AND K562 CELLS WERE CULTURED AND TREATED WITH DEMETHYLATION DRUGS AND HISTONE DEACETYLASE INHIBITORS. REAL TIME QUANTITATIVE POLYMERASE CHAIN REACTION (QPCR), METHYLATION-SPECIFIC PCR, BISULFITE SEQUENCING PCR, WESTERN BLOTTING, CO-IMMUNOPRECIPITATION AND CHROMATIN IMMUNOPRECIPITATION (CHIP) WAS PERFORMED. PTPN6 WAS DOWN-REGULATED IN CELL LINES AND PATIENTS WITH ADVANCED PHASE CML, WHEREAS DNMT1, DNMT3A, MECP2, MBD2 AND HDAC1 WERE UP-REGULATED. TREATMENT WITH 5-AZACYTIDINE, DECITABINE, SODIUM VALPROATE AND LBH589 INCREASED PTPN6 EXPRESSION, BUT DECREASED THAT OF DNMT1, DNMT3A, MECP2, MBD2 AND HDAC1. IMMUNOPRECIPITATION AND MASS SPECTROMETRY SHOWED THAT HDAC1 COMBINED DIRECTLY WITH PTPN6. CHIP-SEQ SHOWED THAT HDAC1 DID NOT COMBINE WITH THE PROMOTER REGION OF PTPN6, WHILE MAPK, AKT, STAT5, JAK2 AND MYC PROMOTER REGIONS ALL COMBINED WITH HDAC1. PTPN6 IS ASSOCIATED WITH PROGRESSION OF CML. LOW EXPRESSION LEVEL OF PTPN6 WAS ASSOCIATED WITH DNA METHYLATION AND REGULATED BY HISTONE ACETYLATION. HDAC1 PARTICIPATES IN THE REGULATION OF PTPN6.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
20  3810 28 INTRATHECAL 5-AZACYTIDINE INHIBITS GLOBAL DNA METHYLATION AND METHYL- CPG-BINDING PROTEIN 2 EXPRESSION AND ALLEVIATES NEUROPATHIC PAIN IN RATS FOLLOWING CHRONIC CONSTRICTION INJURY. THE PATHOGENESIS OF NEUROPATHIC PAIN REMAINS LARGELY UNKNOWN. EPIGENETIC MECHANISMS MAY PLAY A MAJOR ROLE IN REGULATING EXPRESSION OF PRO- OR ANTINOCICEPTIVE GENES. DNA METHYLATION IS A MAJOR EPIGENETIC MECHANISM IN VERTEBRATES, AND METHYL- CPG-BINDING PROTEIN 2 (MECP2) IS DIRECTLY INVOLVED IN METHYLATION-MEDIATED GENE SILENCING. TO DETERMINE HOW CHANGES IN GLOBAL DNA METHYLATION AND MECP2 EXPRESSION OCCUR FOLLOWING CHRONIC CONSTRICTION INJURY (CCI) AND HOW REPRESSION OF DNA METHYLATION AFFECTS THESE CHANGES AND ATTENUATES NEUROPATHIC PAIN, WE USED INTRATHECAL 5-AZACYTIDINE, A DNA METHYLTRANSFERASE INHIBITOR, IN CCI RATS. RATS RECEIVED 0.9% SALINE OR 5-AZACYTIDINE (10MUMOL.D(-1)) VIA SPINAL INJECTION ONCE DAILY FROM DAY 3 TO DAY 14 AFTER CCI SURGERY. GLOBAL DNA METHYLATION AND MECP2 EXPRESSION INCREASED IN THE SPINAL CORD IN CCI RATS ON DAY 14 AFTER CCI SURGERY. MECHANICAL ALLODYNIA AND THERMAL HYPERALGESIA INDUCED BY CCI WERE ATTENUATED BY INTRATHECAL 5-AZACYTIDINE FROM DAY 5 TO DAY 14 AFTER CCI SURGERY. THE INCREASES IN GLOBAL DNA METHYLATION AND MECP2 EXPRESSION IN THE SPINAL CORD IN CCI RATS WERE ALSO SIGNIFICANTLY INHIBITED BY INTRATHECAL 5-AZACYTIDINE. THESE RESULTS DEMONSTRATE THAT INCREASED GLOBAL DNA METHYLATION AND MECP2 EXPRESSION IN THE SPINAL CORD AFTER NERVE DAMAGE MAY PLAY AN IMPORTANT ROLE IN NEUROPATHIC PAIN. 5-AZACYTIDINE SHOWS POTENTIAL FOR TREATING NEUROPATHIC PAIN.	2011