1 2041 129 EPIGENETIC CHANGES WITHIN THE ANNULUS FIBROSUS BY DNA METHYLATION IN RAT INTERVERTEBRAL DISC DEGENERATION MODEL. INTERVERTEBRAL DISC DEGENERATION (IDD) IS AN AGE-DEPENDENT PROGRESSIVE SPINAL DISEASE THAT CAUSES CHRONIC BACK OR NECK PAIN. ALTHOUGH AGING HAS LONG BEEN PRESENTED AS THE MAIN RISK FACTOR, THE EXACT CAUSE IS NOT FULLY KNOWN. DNA METHYLATION IS ASSOCIATED WITH CHRONIC PAIN, SUGGESTING THAT EPIGENETIC MODULATION MAY AMELIORATE DISC DEGENERATION. WE EXAMINED HISTOLOGICAL CHANGES IN THE DNA METHYLATION WITHIN THE DISCS AND THEIR ASSOCIATION WITH PAIN-RELATED TRANSIENT RECEPTOR POTENTIAL VANILLOID SUBTYPE 1 (TRPV1) EXPRESSION IN RATS SUBJECTED TO IDD. EPIGENETIC MARKERS (5-HYDROXYMETHYLCYTOSINE (5HMC), 5-METHYLCYTOSINE (5MC)), DNA METHYLTRANSFERASES (DNMTS), AND TEN-ELEVEN TRANSLOCATIONS (TETS) WERE ANALYZED USING IMMUNOHISTOCHEMISTRY, REAL-TIME PCR, AND DNA DOT-BLOT FOLLOWING IDD. RESULTS REVEALED HIGH 5MC LEVELS IN THE ANNULUS FIBROSUS (AF) REGION WITHIN THE DISC AFTER IDD AND AN ASSOCIATION WITH TRPV1 EXPRESSION. DNMT1 IS MAINLY INVOLVED IN 5MC CONVERSION IN DEGENERATED DISCS. HOWEVER, 5HMC LEVELS DID NOT DIFFER BETWEEN GROUPS. A DEGENERATED DISC CAN LEAD TO LOCOMOTOR DEFECTS AS ASSESSED BY LADDER AND TAIL SUSPENSION TESTS, NO PAIN SIGNALS IN THE VON FREY TEST, UPREGULATED MATRIX METALLOPROTEINASE-3, AND DOWNREGULATED AGGRECAN LEVELS WITHIN THE DISC. THUS, WE FOUND THAT THE DNA METHYLATION STATUS IN THE AF REGION OF THE DISC WAS MAINLY CHANGED AFTER IDD AND ASSOCIATED WITH ABERRANT TRPV1 EXPRESSION IN DEGENERATED DISCS.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
2 2297  29 EPIGENETIC REGULATION OF ACUTE INFLAMMATORY PAIN. ACUTE PAIN IS ASSOCIATED WITH TISSUE DAMAGE, WHICH RESULTS IN THE RELEASE OF INFLAMMATORY MEDIATORS. RECENT STUDIES POINT TO THE INVOLVEMENT OF EPIGENETIC MECHANISMS (DNA METHYLATION) IN THE DEVELOPMENT OF PAIN. WE HAVE FOUND THAT DURING ACUTE INFLAMMATORY PAIN INDUCED BY THE APPLICATION OF 10% MUSTARD OIL ON THE TONGUES OF RATS, LEVELS OF DNMT3A AND 3B WERE ELEVATED MARKEDLY (36 AND 42 % RESPECTIVELY), WHEREAS THE LEVEL OF DNMT1 WAS NOT CHANGED SIGNIFICANTLY. PREVIOUS INJECTION OF XEFOCAM WITH 0,4 MG/KG DOSE DECREASED LEVELS OF DNMT3A AND 3B (25 AND 24% RESPECTIVELY). THE LEVEL OF DNMT1 WAS NOT CHANGED SIGNIFICANTLY COMPARED TO THE CONTROL GROUP. THE FINDINGS SUPPORT THE IDEA THAT INHIBITORS OF DNA-METHYLTRANSFERASES COULD BE USEFUL FOR PAIN MANAGEMENT. OUR DATA SUGGEST THAT NSAIDS (ALONE OR IN COMBINATION WITH DNMT INHIBITORS) MAY BE PROPOSED AS POSSIBLE EPIGENETIC REGULATORY AGENTS, WHICH MAY PLAY A ROLE IN EPIGENETIC MECHANISMS INDIRECTLY THROUGH ALTERING THE ACTIVITY OF INFLAMMATORY MEDIATORS INVOLVED IN PAIN DEVELOPMENT.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
3 6419  33 THE TET2-UPF1 COMPLEX MODULATES MRNA STABILITY UNDER STRESS CONDITIONS. INTRODUCTION: ENVIRONMENTAL STRESS PROMOTES EPIGENETIC ALTERATIONS THAT IMPACT GENE EXPRESSION AND SUBSEQUENTLY PARTICIPATE IN THE PATHOLOGICAL PROCESSES OF THE DISORDER. AMONG EPIGENETIC REGULATIONS, TEN-ELEVEN TRANSLOCATION (TET) ENZYMES OXIDIZE 5-METHYLCYTOSINE (5MC) TO 5-HYDROXYMETHYLCYTOSINE (5HMC) IN DNA AND RNA AND FUNCTION AS CRITICAL PLAYERS IN THE PATHOGENESIS OF DISEASES. OUR PREVIOUS RESULTS SHOWED THAT CHRONIC STRESS INCREASES THE EXPRESSION OF CYTOPLASMIC TET2 IN THE HIPPOCAMPUS OF MICE EXPOSED TO CHRONIC MILD STRESS (CMS). WHETHER THE CYTOPLASMIC TET2 ALTERS RNA 5HMC MODIFICATION IN CHRONIC STRESS-RELATED PROCESSES REMAINS LARGELY UNKNOWN. METHODS: TO EXPLORE THE ROLE OF CYTOPLASMIC TET2 UNDER CMS CONDITIONS, WE ESTABLISHED CMS MICE MODEL AND DETECTED THE EXPRESSION OF RNA 5HMC BY DOT BLOT. WE VERIFIED THE INTERACTION OF TET2 AND ITS INTERACTING PROTEIN BY CO-IMMUNOPRECIPITATION COMBINED WITH MASS SPECTROMETRY AND SCREENED DOWNSTREAM TARGET GENES BY CLUSTER ANALYSIS OF TET2 AND UPSTREAM FRAMESHIFT 1 (UPF1) INTERACTING RNA. THE EXPRESSION OF PROTEIN WAS DETECTED BY WESTERN BLOT AND THE EXPRESSION OF THE SCREENED TARGET GENES WAS DETECTED BY QRT-PCR. RESULTS: IN THIS STUDY, WE FOUND THAT INCREASED CYTOPLASMIC TET2 EXPRESSION UNDER CMS CONDITIONS LEADS TO INCREASE IN TOTAL RNA 5HMC MODIFICATION. TET2 INTERACTED WITH THE KEY NON-SENSE-MEDIATED MRNA DECAY (NMD) FACTOR UPF1, REGULATED THE STABILITY OF STRESS-RELATED GENES SUCH AS UNC5B MRNA, AND MIGHT THEREBY AFFECT NEURODEVELOPMENT. DISCUSSION: IN SUMMARY, THIS STUDY REVEALED THAT TET2-MEDIATED RNA 5HMC MODIFICATION IS INVOLVED IN STRESS-RELATED MRNA STABILITY REGULATION AND MAY SERVE AS A POTENTIAL THERAPEUTIC TARGET FOR CHRONIC STRESS-RELATED DISEASES SUCH AS DEPRESSION.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
4  345  43 ALTERED BRAIN EXPRESSION OF DNA METHYLATION AND HYDROXYMETHYLATION EPIGENETIC ENZYMES IN A RAT MODEL OF NEUROPATHIC PAIN. THE ROLE OF EPIGENETICS IN CHRONIC PAIN AT THE SUPRASPINAL LEVEL IS YET TO BE FULLY CHARACTERIZED. DNA HISTONE METHYLATION IS CRUCIALLY REGULATED BY DE NOVO METHYLTRANSFERASES (DNMT1-3) AND TEN-ELEVEN TRANSLOCATION DIOXYGENASES (TET1-3). EVIDENCE HAS SHOWN THAT METHYLATION MARKERS ARE ALTERED IN DIFFERENT CNS REGIONS RELATED TO NOCICEPTION, NAMELY THE DORSAL ROOT GANGLIA, THE SPINAL CORD, AND DIFFERENT BRAIN AREAS. DECREASED GLOBAL METHYLATION WAS FOUND IN THE DRG, THE PREFRONTAL CORTEX, AND THE AMYGDALA, WHICH WAS ASSOCIATED WITH DECREASED DNMT1/3A EXPRESSION. IN CONTRAST, INCREASED METHYLATION LEVELS AND MRNA LEVELS OF TET1 AND TET3 WERE LINKED TO AUGMENTED PAIN HYPERSENSITIVITY AND ALLODYNIA IN INFLAMMATORY AND NEUROPATHIC PAIN MODELS. SINCE EPIGENETIC MECHANISMS MAY BE RESPONSIBLE FOR THE REGULATION AND COORDINATION OF VARIOUS TRANSCRIPTIONAL MODIFICATIONS DESCRIBED IN CHRONIC PAIN STATES, WITH THIS STUDY, WE AIMED TO EVALUATE THE FUNCTIONAL ROLE OF TET1-3 AND DNMT1/3A GENES IN NEUROPATHIC PAIN IN SEVERAL BRAIN AREAS. IN A SPARED NERVE INJURY RAT MODEL OF NEUROPATHIC PAIN, 21 DAYS AFTER SURGERY, WE FOUND INCREASED TET1 EXPRESSION IN THE MEDIAL PREFRONTAL CORTEX AND DECREASED EXPRESSION IN THE CAUDATE-PUTAMEN AND THE AMYGDALA; TET2 WAS UPREGULATED IN THE MEDIAL THALAMUS; TET3 MRNA LEVELS WERE REDUCED IN THE MEDIAL PREFRONTAL CORTEX AND THE CAUDATE-PUTAMEN; AND DNMT1 WAS DOWNREGULATED IN THE CAUDATE-PUTAMEN AND THE MEDIAL THALAMUS. NO STATISTICALLY SIGNIFICANT CHANGES IN EXPRESSION WERE OBSERVED WITH DNMT3A. OUR RESULTS SUGGEST A COMPLEX FUNCTIONAL ROLE FOR THESE GENES IN DIFFERENT BRAIN AREAS IN THE CONTEXT OF NEUROPATHIC PAIN. THE NOTION OF DNA METHYLATION AND HYDROXYMETHYLATION BEING CELL-TYPE SPECIFIC AND NOT TISSUE SPECIFIC, AS WELL AS THE POSSIBILITY OF CHRONOLOGICALLY DIFFERENTIAL GENE EXPRESSION AFTER THE ESTABLISHMENT OF NEUROPATHIC OR INFLAMMATORY PAIN MODELS, OUGHT TO BE ADDRESSED IN FUTURE STUDIES.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
5  277  33 AGE-RELATED EPIGENOME-WIDE DNA METHYLATION AND HYDROXYMETHYLATION IN LONGITUDINAL MOUSE BLOOD. DNA METHYLATION AT CYTOSINE-PHOSPHATE-GUANINE (CPG) DINUCLEOTIDES CHANGES AS A FUNCTION OF AGE IN HUMANS AND ANIMAL MODELS, A PROCESS THAT MAY CONTRIBUTE TO CHRONIC DISEASE DEVELOPMENT. RECENT STUDIES HAVE INVESTIGATED THE ROLE OF AN OXIDIZED FORM OF DNA METHYLATION - 5-HYDROXYMETHYLCYTOSINE (5HMC) - IN THE EPIGENOME, BUT ITS CONTRIBUTION TO AGE-RELATED DNA METHYLATION REMAINS UNCLEAR. WE TESTED THE HYPOTHESIS THAT 5HMC CHANGES WITH AGE, BUT IN A DIRECTION OPPOSITE TO 5-METHYLCYTOSINE (5MC), POTENTIALLY PLAYING A DISTINCT ROLE IN AGING. TO CHARACTERIZE EPIGENETIC AGING, GENOME-WIDE 5MC AND 5HMC WERE MEASURED IN LONGITUDINAL BLOOD SAMPLES (2, 4, AND 10 MONTHS OF AGE) FROM ISOGENIC MICE USING TWO SEQUENCING METHODS - ENHANCED REDUCED REPRESENTATION BISULFITE SEQUENCING AND HYDROXYMETHYLATED DNA IMMUNOPRECIPITATION SEQUENCING. EXAMINING THE EPIGENOME BY AGE, WE IDENTIFIED 28,196 UNIQUE DIFFERENTIALLY METHYLATED CPGS (DMCS) AND 8,613 DIFFERENTIALLY HYDROXYMETHYLATED REGIONS (DHMRS). MOUSE BLOOD SHOWED A GENERAL PATTERN OF EPIGENOME-WIDE HYPERMETHYLATION AND HYPO-HYDROXYMETHYLATION WITH AGE. COMPARING AGE-RELATED DMCS AND DHMRS, 1,854 ANNOTATED GENES SHOWED BOTH DIFFERENTIAL 5MC AND 5HMC, INCLUDING ONE GENE - NFIC - AT FIVE CPGS IN THE SAME 250 BP CHROMOSOMAL REGION. AT THIS REGION, 5MC AND 5HMC LEVELS BOTH DECREASED WITH AGE. REFLECTING THESE AGE-RELATED EPIGENETIC CHANGES, NFIC RNA EXPRESSION IN BLOOD DECREASED WITH AGE, SUGGESTING THAT AGE-RELATED REGULATION OF THIS GENE MAY BE DRIVEN BY 5HMC, NOT CANONICAL DNA METHYLATION. COMBINED, OUR GENOME-WIDE RESULTS SHOW AGE-RELATED DIFFERENTIAL 5MC AND 5HMC, AS WELL AS SOME EVIDENCE THAT CHANGES IN 5HMC MAY DRIVE AGE-RELATED DNA METHYLATION AND GENE EXPRESSION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
6 5972  28 TET REPRESSION AND INCREASED DNMT ACTIVITY SYNERGISTICALLY INDUCE ABERRANT DNA METHYLATION. CHRONIC INFLAMMATION IS DEEPLY INVOLVED IN VARIOUS HUMAN DISORDERS, SUCH AS CANCER, NEURODEGENERATIVE DISORDERS, AND METABOLIC DISORDERS. INDUCTION OF EPIGENETIC ALTERATIONS, ESPECIALLY ABERRANT DNA METHYLATION, IS ONE OF THE MAJOR MECHANISMS, BUT HOW IT IS INDUCED IS STILL UNCLEAR. HERE, WE FOUND THAT EXPRESSION OF TET GENES, METHYLATION ERASERS, WAS DOWNREGULATED IN INFLAMED MOUSE AND HUMAN TISSUES, AND THAT THIS WAS CAUSED BY UPREGULATION OF TET-TARGETING MIRNAS SUCH AS MIR20A, MIR26B, AND MIR29C, LIKELY DUE TO ACTIVATION OF NF-KAPPAB SIGNALING DOWNSTREAM OF IL-1BETA AND TNF-ALPHA. HOWEVER, TET KNOCKDOWN INDUCED ONLY MILD ABERRANT METHYLATION. NITRIC OXIDE (NO), PRODUCED BY NOS2, ENHANCED ENZYMATIC ACTIVITY OF DNA METHYLTRANSFERASES (DNMTS), METHYLATION WRITERS, AND NO EXPOSURE INDUCED MINIMAL ABERRANT METHYLATION. IN CONTRAST, A COMBINATION OF TET KNOCKDOWN AND NO EXPOSURE SYNERGISTICALLY INDUCED ABERRANT METHYLATION, INVOLVING GENOMIC REGIONS NOT METHYLATED BY EITHER ALONE. THE RESULTS SHOWED THAT A VICIOUS COMBINATION OF TET REPRESSION, DUE TO NF-KAPPAB ACTIVATION, AND DNMT ACTIVATION, DUE TO NO PRODUCTION, IS RESPONSIBLE FOR ABERRANT METHYLATION INDUCTION IN HUMAN TISSUES.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
7  226  34 ACUTE TRANSCRIPTOMIC AND EPIGENETIC ALTERATIONS AT T12 AFTER RAT T10 SPINAL CORD CONTUSIVE INJURY. SPINAL CORD INJURY IS A SEVERELY DEBILITATING CONDITION AFFECTING A SIGNIFICANT POPULATION IN THE USA. SPINAL CORD INJURY PATIENTS OFTEN HAVE INCREASED RISK OF DEVELOPING PERSISTENT NEUROPATHIC PAIN AND OTHER NEURODEGENERATIVE CONDITIONS BEYOND THE PRIMARY LESION CENTER LATER IN THEIR LIFE. THE MOLECULAR MECHANISM CONFERRING TO THE "LATENT" DAMAGES AT DISTAL TISSUES, HOWEVER, REMAINS ELUSIVE. HERE, WE STUDIED MOLECULAR CHANGES CONFERRING ABNORMAL FUNCTIONALITY AT DISTAL SPINAL CORD (T12) BEYOND THE LESION CENTER (T10) BY COMBINING NEXT-GENERATION SEQUENCING (RNA- AND BISULFITE SEQUENCING), SUPER-RESOLUTION MICROSCOPY, AND IMMUNOFLUORESCENCE STAINING AT 7 DAYS POST INJURY. WE OBSERVED SIGNIFICANT TRANSCRIPTOMIC CHANGES PRIMARILY ENRICHED IN NEUROINFLAMMATION AND SYNAPTOGENESIS ASSOCIATED PATHWAYS. TRANSCRIPTION FACTORS (TFS) THAT REGULATE NEUROGENESIS AND NEURON PLASTICITY, INCLUDING EGR1, KLF4, AND MYC, ARE SIGNIFICANTLY UPREGULATED. ALONG WITH GLOBAL CHANGES IN CHROMATIN ARRANGEMENTS AND DNA METHYLATION, INCLUDING 5-METHYLCYTOSINE (5MC) AND 5-HYDROXYMETHYLCYTOSINE (5HMC), BISULFITE SEQUENCING FURTHER REVEALS THE INVOLVEMENT OF DNA METHYLATION CHANGES IN REGULATING CYTOKINE, GROWTH FACTOR, AND ION CHANNEL EXPRESSION. COLLECTIVELY, OUR RESULTS PAVE THE WAY TOWARDS UNDERSTANDING TRANSCRIPTOMIC AND EPIGENOMIC MECHANISM IN CONFERRING LONG-TERM DISEASE RISKS AT DISTAL TISSUES AWAY FROM THE PRIMARY LESION CENTER AND SHED LIGHT ON POTENTIAL MOLECULAR TARGETS THAT GOVERN THE REGULATORY MECHANISM AT DISTAL SPINAL CORD TISSUES.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
8 3729  39 INHIBITION OF TET1 PREVENTS THE DEVELOPMENT OF OSTEOARTHRITIS AND REVEALS THE 5HMC LANDSCAPE THAT ORCHESTRATES PATHOGENESIS. OSTEOARTHRITIS (OA) IS A DEGENERATIVE DISEASE OF THE JOINT, WHICH RESULTS IN PAIN, LOSS OF MOBILITY, AND, EVENTUALLY, JOINT REPLACEMENT. CURRENTLY, NO DISEASE-MODIFYING DRUGS EXIST, PARTLY BECAUSE OF THE MULTIPLE LEVELS AT WHICH CARTILAGE HOMEOSTASIS IS DISRUPTED. RECENT STUDIES HAVE HIGHLIGHTED THE IMPORTANCE OF EPIGENETIC DYSREGULATION IN OA, SPARKING INTEREST IN THE EPIGENETIC MODULATION FOR THIS DISEASE. IN OUR PREVIOUS WORK, WE CHARACTERIZED A FIVEFOLD INCREASE IN CYTOSINE HYDROXYMETHYLATION (5HMC), AN OXIDIZED DERIVATIVE OF CYTOSINE METHYLATION (5MC) ASSOCIATED WITH GENE ACTIVATION, ACCUMULATING AT OA-ASSOCIATED GENES. TO TEST THE ROLE OF 5HMC IN OA, HERE, WE USED A MOUSE MODEL OF SURGICALLY INDUCED OA AND FOUND THAT OA ONSET WAS ACCOMPANIED BY A GAIN OF ~40,000 DIFFERENTIALLY HYDROXYMETHYLATED SITES BEFORE THE NOTABLE HISTOLOGICAL APPEARANCE OF DISEASE. WE DEMONSTRATED THAT TEN-ELEVEN-TRANSLOCATION ENZYME 1 (TET1) MEDIATES THE 5HMC DEPOSITION BECAUSE 98% OF SITES ENRICHED FOR 5HMC IN OA WERE LOST IN TET1(-/-) MICE. LOSS OF TET1-MEDIATED 5HMC PROTECTED THE TET1(-/-) MICE FROM OA DEVELOPMENT, INCLUDING DEGENERATION OF THE CARTILAGE SURFACE AND OSTEOPHYTE FORMATION, BY DIRECTLY PREVENTING THE ACTIVATION OF MULTIPLE OA PATHWAYS. LOSS OF TET1 IN HUMAN OA CHONDROCYTES REDUCED THE EXPRESSION OF THE MATRIX METALLOPROTEINASES MMP3 AND MMP13 AND MULTIPLE INFLAMMATORY CYTOKINES. INTRA-ARTICULAR INJECTIONS OF A DIOXYGENASES INHIBITOR, 2-HYDROXYGLUTARATE, ON MICE AFTER SURGICAL INDUCTION OF OA STALLED DISEASE PROGRESSION. TREATMENT OF HUMAN OA CHONDROCYTES WITH THE SAME INHIBITOR ALSO PHENOCOPIED TET1 LOSS. COLLECTIVELY, THESE DATA DEMONSTRATE THAT TET1-MEDIATED 5HMC DEPOSITION REGULATES MULTIPLE OA PATHWAYS AND CAN BE MODULATED FOR THERAPEUTIC INTERVENTION.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
9  839  30 CHEMO-ENZYMATIC FLUORESCENCE LABELING OF GENOMIC DNA FOR SIMULTANEOUS DETECTION OF GLOBAL 5-METHYLCYTOSINE AND 5-HYDROXYMETHYLCYTOSINE. 5-METHYLCYTOSINE AND 5-HYDROXYMETHYLCYTOSINE ARE EPIGENETIC MODIFICATIONS INVOLVED IN GENE REGULATION AND CANCER. WE PRESENT A NEW, SIMPLE, AND HIGH-THROUGHPUT PLATFORM FOR MULTI-COLOR EPIGENETIC ANALYSIS. THE NOVELTY OF OUR APPROACH IS THE ABILITY TO MULTIPLEX METHYLATION AND DE-METHYLATION SIGNALS IN THE SAME ASSAY. WE UTILIZE AN ENGINEERED METHYLTRANSFERASE ENZYME THAT RECOGNIZES AND LABELS ALL UNMODIFIED CPG SITES WITH A FLUORESCENT COFACTOR. IN COMBINATION WITH THE ALREADY ESTABLISHED LABELING OF THE DE-METHYLATION MARK 5-HYDROXYMETHYLCYTOSINE VIA ENZYMATIC GLYCOSYLATION, WE OBTAINED A ROBUST PLATFORM FOR SIMULTANEOUS EPIGENETIC ANALYSIS OF THESE MARKS. WE ASSESSED THE GLOBAL EPIGENETIC LEVELS IN MULTIPLE SAMPLES OF COLORECTAL CANCER AND OBSERVED A 3.5-FOLD REDUCTION IN 5HMC LEVELS BUT NO CHANGE IN DNA METHYLATION LEVELS BETWEEN SICK AND HEALTHY INDIVIDUALS. WE ALSO MEASURED EPIGENETIC MODIFICATIONS IN CHRONIC LYMPHOCYTIC LEUKEMIA AND OBSERVED A DECREASE IN BOTH MODIFICATION LEVELS (5-HYDROXYMETHYLCYTOSINE: WHOLE BLOOD 30 %; PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) 40 %. 5-METHYLCYTOSINE: WHOLE BLOOD 53 %; PBMCS 48 %). OUR FINDINGS PROPOSE USING A SIMPLE BLOOD TEST AS A VIABLE METHOD FOR ANALYSIS, SIMPLIFYING SAMPLE HANDLING IN DIAGNOSTICS. IMPORTANTLY, OUR RESULTS HIGHLIGHT THE ASSAY'S POTENTIAL FOR EPIGENETIC EVALUATION OF CLINICAL SAMPLES, BENEFITING RESEARCH AND PATIENT MANAGEMENT.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
10 5975  21 TET1 IS AN IMPORTANT TRANSCRIPTIONAL ACTIVATOR OF TNFALPHA EXPRESSION IN MACROPHAGES. ACTIVATION OF MACROPHAGES AND OVEREXPRESSION OF TNFALPHA IS ASSOCIATED WITH THE PATHOGENESIS OF CHRONIC INFLAMMATORY DISEASES. HOWEVER, THE MECHANISMS LEADING TO TNFALPHA OVEREXPRESSION ARE STILL UNKNOWN. 5-METHYLOCYTOSINE (5-MC) IS AN EPIGENETIC MODIFICATION THAT IS ASSOCIATED WITH SILENCED GENES. RECENT STUDIES SHOWED THAT IT IS CONVERTED TO 5-HYDROXYLMETHYLOCYTOSINE (5-HMC) AND REACTIVATES GENE EXPRESSION THROUGH THE ACTION OF THE FAMILY OF TEN-ELEVEN-TRANSLOCATION (TET1-3) ENZYMES. IN THIS STUDY, WE SHOW THAT 5-HMC LEVELS ARE INCREASED GLOBALLY AND SPECIFICALLY IN THE TNFALPHA PROMOTER DURING THE DIFFERENTIATION OF MONOCYTES TO MACROPHAGES. IN ADDITION, THE LEVELS OF 5-HMC ARE INCREASED UPON LPS STIMULATION OF MACROPHAGES. FURTHERMORE, CRIPSR STABLE KNOCKOUT OF TET1 DECREASES THE EXPRESSION OF TNFALPHA AND OTHER PRO-INFLAMMATORY CYTOKINES. IN CONCLUSION, WE SHOWED THAT TET1 CONTRIBUTES TO THE ACTIVATION OF MACROPHAGES POSSIBLY THROUGH REGULATION OF 5-HYDROXYMETHYLATION IN THE PROMOTER OF PRO-INFLAMMATORY CYTOKINE GENES. THE TET1 ENZYME COULD BE A PROMISING THERAPEUTIC TARGET TO INHIBIT THE PERSISTENT INFLAMMATION CAUSED BY MACROPHAGES IN CHRONIC INFLAMMATORY DISEASES.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
11 2762  37 EXPRESSION OF TET2 ENZYME INDICATES ENHANCED EPIGENETIC MODIFICATION OF CELLS IN PERIODONTITIS. DNA METHYLATION IS AN IMPORTANT EPIGENETIC MECHANISM INVOLVED IN THE REGULATION OF GENE EXPRESSION, AND A REDUCTION IN DNA METHYLATION INFLUENCES CELL-CYCLE PROGRESSION AND CELL DIFFERENTIATION IN INFLAMMATORY CELLS. THE AIM OF THE PRESENT STUDY WAS TO ANALYZE THE DNA-METHYLATION PATTERN AT LOCAL AND GLOBAL/SYSTEMIC LEVELS IN PATIENTS WITH PERIODONTITIS AND GINGIVITIS. TWENTY-ONE SUBJECTS WITH GENERALIZED, SEVERE PERIODONTITIS AND 17 SUBJECTS WITH GINGIVAL INFLAMMATION BUT NO ATTACHMENT LOSS WERE RECRUITED. GINGIVAL BIOPSIES AND PERIPHERAL BLOOD SAMPLES WERE COLLECTED AND PREPARED FOR IMMUNOHISTOCHEMICAL ANALYSIS OF 5-METHYLCYTOSINE (5MC), 5-HYDROXYMETHYLCYTOSINE (5HMC), TEN-ELEVEN TRANSLOCATION 2 (TET2), AND DNA METHYLTRANSFERASE 1 (DNMT1). WHILST A SIMILAR PATTERN FOR 5MC AND 5HMC DNA METHYLATION WAS FOUND IN BOTH TYPES OF LESIONS, A SIGNIFICANTLY LARGER PROPORTION OF TET2-POSITIVE CELLS WAS FOUND IN PERIODONTITIS LESIONS THAN IN GINGIVITIS LESIONS. QUANTITATIVE REAL-TIME PCR ANALYSIS SHOWED NO DIFFERENCES BETWEEN GINGIVITIS AND PERIODONTITIS LESIONS REGARDING EXPRESSION OF TET2 AND ISOCITRATE DEHYDROGENASE (IDH) GENES, WHILE THE GLOBAL LEVEL OF 5HMC WAS SIGNIFICANTLY HIGHER IN BLOOD THAN IN TISSUE IN PATIENTS WITH PERIODONTITIS. IT IS SUGGESTED THAT EPIGENETIC CHANGES ARE MORE COMMON IN PERIODONTITIS LESIONS THAN IN GINGIVITIS LESIONS AND THAT SUCH CHANGES ARE TISSUE SPECIFIC.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
12 4879  26 OVERLAPPING SIGNATURES OF CHRONIC PAIN IN THE DNA METHYLATION LANDSCAPE OF PREFRONTAL CORTEX AND PERIPHERAL T CELLS. WE TESTED THE HYPOTHESIS THAT EPIGENETIC MECHANISMS IN THE BRAIN AND THE IMMUNE SYSTEM ARE ASSOCIATED WITH CHRONIC PAIN. GENOME-WIDE DNA METHYLATION ASSESSED IN 9 MONTHS POST NERVE-INJURY (SNI) AND SHAM RATS, IN THE PREFRONTAL CORTEX (PFC) AS WELL AS IN T CELLS REVEALED A VAST DIFFERENCE IN THE DNA METHYLATION LANDSCAPE IN THE BRAIN BETWEEN THE GROUPS AND A REMARKABLE OVERLAP (72%) BETWEEN DIFFERENTIALLY METHYLATED PROBES IN T CELLS AND PREFRONTAL CORTEX. DNA METHYLATION STATES IN THE PFC SHOWED ROBUST CORRELATION WITH PAIN SCORE OF ANIMALS IN SEVERAL GENES INVOLVED IN PAIN. FINALLY, ONLY 11 DIFFERENTIALLY METHYLATED PROBES IN T CELLS WERE SUFFICIENT TO DISTINGUISH SNI OR SHAM INDIVIDUAL RATS. THIS STUDY SUPPORTS THE PLAUSIBILITY OF DNA METHYLATION INVOLVEMENT IN CHRONIC PAIN AND DEMONSTRATES THE POTENTIAL FEASIBILITY OF DNA METHYLATION MARKERS IN T CELLS AS NONINVASIVE BIOMARKERS OF CHRONIC PAIN SUSCEPTIBILITY.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
13 1813  36 EFFECTS OF CAFFEIC ACID ON EPIGENETICS IN THE BRAIN OF RATS WITH CHRONIC UNPREDICTABLE MILD STRESS. THE PRESENT STUDY HYPOTHESIZED THAT CAFFEIC ACID (3,4?DIHYDROXYCINNAMIC ACID; CAA) MAY EXERT ANTIDEPRESSANT?LIKE EFFECTS IN RATS WITH CHRONIC UNPREDICTABLE MILD STRESS VIA EPIGENETIC MECHANISMS, SUCH AS DNA METHYLATION AND HYDROXYMETHYLATION. THE CHRONIC UNPREDICTABLE MILD STRESS (CUMS) MODEL WAS USED TO ANALYZE THE EFFECTS OF CAA ON BEHAVIORAL PHENOTYPES, AND TO EVALUATE THE DISTRIBUTION OF 5?METHYLCYTOSINE (5MC) AND 5?HYDROXYMETHYLCYTOSINE (5HMC) IN THE HIPPOCAMPUS AND PREFRONTAL CORTEX USING IMMUNOHISTOCHEMISTRY AND IMMUNOFLUORESCENCE. MRNA LEVELS OF THE GENES ENCODING BRAIN?DERIVED NEUROTROPIC FACTOR (BDNF) AND CATECHOL?O?METHYLTRANSFERASE (COMT), AND KEY ENZYMES REGULATING DNA METHYLATION [DNA METHYLTRANSFERASE (DNMT)1 AND DNMT3A] AND HYDROXYMETHYLATION [TEN?ELEVEN TRANSLOCATION (TET)1?3] WERE EXAMINED USING QUANTITATIVE (Q)PCR. FURTHERMORE, ENRICHMENT OF 5MC AND 5HMC AT THE PROMOTOR REGIONS OF THE BDNF AND COMT GENES WAS QUANTIFIED USING CHROMATIN IMMUNOPRECIPITATION?QPCR. BEHAVIORAL DATA SHOWED THAT CAA EXERTED A SLIGHT ANTIDEPRESSANT?LIKE EFFECT. BDNF AND COMT GENES SHOWED DIFFERENTIAL EXPRESSION PATTERNS DUE TO CUMS. CAA INTERVENTION INDUCED DIFFERENT DNMT1/DNMT3A AND TET1/TET2 MRNA LEVELS IN THE HIPPOCAMPUS AND PREFRONTAL CORTEX, RESPECTIVELY. CAA REGULATED THE RATIO OF 5MC/5HMC AT THE PROMOTOR REGION OF THE BDNF AND COMT GENES AND THEREFORE INFLUENCED GENE EXPRESSION, WHICH MAY BE A VALUABLE THERAPEUTIC OPTION FOR MAJOR DEPRESSIVE DISORDER (MDD). IN CONCLUSION, THERE WERE EPIGENETIC CHANGES IN THE HIPPOCAMPUS AND PREFRONTAL CORTEX IN CUMS RATS, AND CAA MAY FUNCTION AS A MODULATOR OF DNA METHYLATION TO REGULATE GENE TRANSCRIPTION, THUS PROVIDING A MECHANISTIC BASIS FOR THE USE OF THIS PHYTOCHEMICAL AGENT IN THE TREATMENT OF MDD.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
14  920  24 CHRONIC HYPOXIA FACILITATES ALZHEIMER'S DISEASE THROUGH DEMETHYLATION OF GAMMA-SECRETASE BY DOWNREGULATING DNA METHYLTRANSFERASE 3B. INTRODUCTION: ENVIRONMENTAL FACTORS AND EPIGENETIC MECHANISMS ARE BELIEVED TO CONTRIBUTE TO ALZHEIMER'S DISEASE (AD). WE PREVIOUSLY DOCUMENTED THAT PRENATAL HYPOXIA AGGRAVATED THE COGNITIVE IMPAIRMENT AND NEUROPATHOLOGY IN OFFSPRING MICE. HERE, WE INVESTIGATE THE CHRONIC HYPOXIA-INDUCED EPIGENETIC MODIFICATIONS IN AD. METHODS: THE 3-MONTH-OLD APP(SWE)/PS1(DE9) MICE WERE EXPOSED TO HYPOXIC ENVIRONMENT 6 HOUR/DAY FOR 30 DAYS, FOLLOWED BY LEARNING AND MEMORY TESTS AND BIOCHEMICAL AND NEUROPATHOLOGY MEASUREMENT AT THE AGE OF 4, 6, AND 9 MONTHS. RESULTS: WE FOUND HYPOXIA EXAGGERATED THE NEUROPATHOLOGY AND COGNITIVE IMPAIRMENT IN AD MICE. CHRONIC HYPOXIA INDUCED DEMETHYLATION ON GENOMIC DNA AND DECREASED THE EXPRESSION OF DNA METHYLTRANSFERASE 3B (DNMT3B) IN VIVO. WE FURTHER FOUND THAT DNMTS INHIBITION ELEVATED THE PROTEIN LEVELS OF AMYLOID PRECURSOR PROTEIN, BETA- AND GAMMA-SECRETASES, WHEREAS OVEREXPRESSION OF DNMT3B SUPPRESSED THE LEVELS OF THEM IN VITRO. DISCUSSION: OUR STUDY SUGGESTS CHRONIC HYPOXIA CAN AGGRAVATE AD PROGRESSION THROUGH DEMETHYLATION OF GENES ENCODING GAMMA-SECRETASE COMPONENTS BY DOWNREGULATION OF DNMT3B.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
15 6246  30 THE METHYL DONOR S-ADENOSYL METHIONINE REVERSES THE DNA METHYLATION SIGNATURE OF CHRONIC NEUROPATHIC PAIN IN MOUSE FRONTAL CORTEX. CHRONIC PAIN IS ASSOCIATED WITH PERSISTENT BUT REVERSIBLE STRUCTURAL AND FUNCTIONAL CHANGES IN THE PREFRONTAL CORTEX (PFC). THIS STABLE YET MALLEABLE PLASTICITY IMPLICATES EPIGENETIC MECHANISMS, INCLUDING DNA METHYLATION, AS A POTENTIAL MEDIATOR OF CHRONIC PAIN-INDUCED CORTICAL PATHOLOGY. WE PREVIOUSLY DEMONSTRATED THAT CHRONIC ORAL ADMINISTRATION OF THE METHYL DONOR S-ADENOSYL METHIONINE (SAM) ATTENUATES LONG-TERM PERIPHERAL NEUROPATHIC PAIN AND ALTERS GLOBAL FRONTAL CORTICAL DNA METHYLATION. HOWEVER, THE SPECIFIC GENES AND PATHWAYS ASSOCIATED WITH THE RESOLUTION OF CHRONIC PAIN BY SAM REMAIN UNEXPLORED. OBJECTIVE: TO DETERMINE THE EFFECT OF LONG-TERM THERAPEUTIC EXPOSURE TO SAM ON THE DNA METHYLATION OF INDIVIDUAL GENES AND PATHWAYS IN A MOUSE NEUROPATHIC PAIN MODEL. METHODS: MALE CD-1 MICE RECEIVED SPARED NERVE INJURY OR SHAM SURGERY. THREE MONTHS AFTER INJURY, ANIMALS RECEIVED SAM (20 MG/KG, ORAL, 3X A WEEK) OR VEHICLE FOR 16 WEEKS FOLLOWED BY EPIGENOME-WIDE ANALYSIS OF FRONTAL CORTEX. RESULTS: PERIPHERAL NEUROPATHIC PAIN WAS ASSOCIATED WITH 4000 DIFFERENTIALLY METHYLATED GENOMIC REGIONS THAT WERE ENRICHED IN INTRACELLULAR SIGNALING, CELL MOTILITY AND MIGRATION, CYTOSKELETAL STRUCTURE, AND CELL ADHESION PATHWAYS. A THIRD OF THESE DIFFERENTIALLY METHYLATED REGIONS WERE REVERSED BY SAM TREATMENT (1415 REGIONS REPRESENTING 1013 GENES). MORE THAN 100 GENES WITH KNOWN PAIN-RELATED FUNCTION WERE DIFFERENTIALLY METHYLATED AFTER NERVE INJURY; 29 OF THESE WERE REVERSED BY SAM TREATMENT INCLUDING SCN10A, TRPA1, NTRK1, AND GFAP. CONCLUSION: THESE RESULTS SUGGEST A ROLE FOR THE EPIGENOME IN THE MAINTENANCE OF CHRONIC PAIN AND ADVANCE EPIGENETIC MODULATORS SUCH AS SAM AS A NOVEL APPROACH TO TREAT CHRONIC PAIN.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
16 1794  35 EFFECT OF DIABETES STATUS AND HYPERGLYCEMIA ON GLOBAL DNA METHYLATION AND HYDROXYMETHYLATION. TYPE 2 DIABETES MELLITUS (T2DM) IS CHARACTERIZED BY OXIDATIVE STRESS THAT COULD LEAD TO CHRONIC MICRO- AND MACROVASCULAR COMPLICATIONS. WE HYPOTHESIZED THAT SOME OF THE TARGET ORGAN DAMAGE IS MEDIATED BY OXIDATIVE ALTERATIONS IN EPIGENETIC MECHANISMS INVOLVING DNA METHYLATION (5MC) AND DNA HYDROXYMETHYLATION (5HMC). WE ANALYZED GLOBAL DNA METHYLATION AND HYDROXYMETHYLATION IN PERIPHERAL BLOOD CELLS IN WELL-CONTROLLED AND POORLY CONTROLLED PATIENTS WITH T2DM AND COMPARED THEM WITH HEALTHY CONTROLS. WE ALSO ANALYZED MICROARRAYS OF DNA METHYLATION AND GENE EXPRESSION OF OTHER IMPORTANT TISSUES IN THE CONTEXT OF DIABETES FROM THE GEO DATABASE REPOSITORY AND THEN COMPARED THESE RESULTS WITH OUR EXPERIMENTAL GENE EXPRESSION DATA. DNA METHYLATION AND, MORE IMPORTANTLY, DNA HYDROXYMETHYLATION LEVELS WERE INCREASED IN POORLY CONTROLLED PATIENTS COMPARED TO WELL-CONTROLLED AND HEALTHY INDIVIDUALS. BOTH 5MC AND 5HMC MEASUREMENTS WERE CORRELATED WITH THE PERCENTAGE OF GLYCATED HEMOGLOBIN, INDICATING A DIRECT IMPACT OF HYPERGLYCEMIA ON CHANGES OVER THE EPIGENOME. THE ANALYSIS OF METHYLATION MICROARRAYS WAS CONCORDANT, AND 5MC LEVELS WERE INCREASED IN THE PERIPHERAL BLOOD OF T2DM PATIENTS. HOWEVER, THE DNA METHYLATION LEVELS WERE THE OPPOSITE OF THOSE IN OTHER TISSUES, SUCH AS THE PANCREAS, ADIPOSE TISSUE AND SKELETAL MUSCLE. WE HYPOTHESIZE THAT A PROCESS OF DNA OXIDATION ASSOCIATED WITH HYPERGLYCEMIA MAY EXPLAIN THE DNA DEMETHYLATION IN WHICH THE ACTIVITY OF TEN-ELEVEN TRANSLOCATION (TET) PROTEINS IS NOT SUFFICIENT TO COMPLETE THE PROCESS. HIGH LEVELS OF GLUCOSE LEAD TO CELLULAR OXIDATION, WHICH TRIGGERS THE PROCESS OF DNA DEMETHYLATION AIDED BY TET ENZYMES, RESULTING IN EPIGENETIC DYSREGULATION OF THE DAMAGED TISSUES.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
17 2706  49 EXERCISE ATTENUATES LOW BACK PAIN AND ALTERS EPIGENETIC REGULATION IN INTERVERTEBRAL DISCS IN A MOUSE MODEL. BACKGROUND CONTEXT: CHRONIC LOW BACK PAIN (LBP) IS A MULTIFACTORIAL DISORDER WITH COMPLEX UNDERLYING MECHANISMS, INCLUDING ASSOCIATIONS WITH INTERVERTEBRAL DISC (IVD) DEGENERATION IN SOME INDIVIDUALS. IT HAS BEEN DEMONSTRATED THAT EPIGENETIC PROCESSES ARE INVOLVED IN THE PATHOLOGY OF IVD DEGENERATION. EPIGENETICS REFERS TO SEVERAL MECHANISMS, INCLUDING DNA METHYLATION, THAT HAVE THE ABILITY TO CHANGE GENE EXPRESSION WITHOUT INDUCING ANY CHANGE IN THE UNDERLYING DNA SEQUENCE. DNA METHYLATION CAN ALTER THE ENTIRE STATE OF A TISSUE FOR AN EXTENDED PERIOD OF TIME AND THUS COULD POTENTIALLY BE HARNESSED FOR LONG-TERM PAIN RELIEF. LIFESTYLE FACTORS, SUCH AS PHYSICAL ACTIVITY, HAVE A STRONG INFLUENCE ON EPIGENETIC REGULATION. EXERCISE IS A COMMONLY PRESCRIBED TREATMENT FOR CHRONIC LBP, AND SEX-SPECIFIC EPIGENETIC ADAPTATIONS IN RESPONSE TO ENDURANCE EXERCISE HAVE BEEN REPORTED. HOWEVER, WHETHER EXERCISE INTERVENTIONS THAT ATTENUATE LBP ARE ASSOCIATED WITH EPIGENETIC ALTERATIONS IN DEGENERATING IVDS HAS NOT BEEN EVALUATED. PURPOSE: WE HYPOTHESIZE THAT THE THERAPEUTIC EFFICACY OF PHYSICAL ACTIVITY IS MEDIATED, AT LEAST IN PART, AT THE EPIGENETIC LEVEL. THE PURPOSE OF THIS STUDY WAS TO USE THE SPARC-NULL MOUSE MODEL OF LBP ASSOCIATED WITH IVD DEGENERATION TO CLARIFY (1) IF IVD DEGENERATION IS ASSOCIATED WITH ALTERED EXPRESSION OF EPIGENETIC REGULATORY GENES IN THE IVDS, (2) IF EPIGENETIC REGULATORY MACHINERY IS SENSITIVE TO THERAPEUTIC ENVIRONMENTAL INTERVENTION, AND (3) IF THERE ARE SEX-SPECIFIC DIFFERENCES IN (1) AND/OR (2). STUDY DESIGN: EIGHT-MONTH-OLD MALE AND FEMALE SPARC-NULL AND AGE-MATCHED CONTROL (WT) MICE (N=108) WERE ASSIGNED TO EXERCISE (N=56) OR SEDENTARY (N=52) GROUPS. DELETION OF SPARC IS ASSOCIATED WITH PROGRESSIVE IVD DEGENERATION AND BEHAVIORAL SIGNS OF LBP. THE EXERCISE GROUP RECEIVED A CIRCULAR PLASTIC HOME CAGE RUNNING WHEEL ON WHICH THEY COULD RUN FREELY. THE SEDENTARY GROUP RECEIVED AN IDENTICAL WHEEL SECURED IN PLACE TO PREVENT ROTATION. AFTER 6 MONTHS, THE RESULTS OBTAINED IN EACH GROUP WERE COMPARED. METHODS: AFTER 6 MONTHS OF EXERCISE, LBP-RELATED BEHAVIORAL INDICES WERE DETERMINED, AND GLOBAL DNA METHYLATION (5-METHYLCYTOSINE) AND EPIGENETIC REGULATORY GENE MRNA EXPRESSION IN IVDS WERE ASSESSED. THIS PROJECT WAS SUPPORTED BY THE CANADIAN INSTITUTES FOR HEALTH RESEARCH. THE AUTHORS HAVE NO CONFLICTS OF INTEREST. RESULTS: LUMBAR IVDS FROM WT SEDENTARY AND SPARC-NULL SEDENTARY MICE HAD SIMILAR LEVELS OF GLOBAL DNA METHYLATION (%5-MC) AND COMPARABLE MRNA EXPRESSION OF EPIGENETIC REGULATORY GENES (DNMT1,3A,B, MECP2, MBD2A,B, TET1-3) IN BOTH SEXES. EXERCISE ATTENUATED LBP-RELATED BEHAVIORS, DECREASED GLOBAL DNA METHYLATION IN BOTH WT (P<.05) AND SPARC-NULL MICE (P<.01) AND REDUCED MRNA EXPRESSION OF MECP2 IN SPARC-NULL MICE (P<.05). SEX-SPECIFIC EFFECTS OF EXERCISE ON EXPRESSION OF MRNA WERE ALSO OBSERVED. CONCLUSIONS: EXERCISE ALLEVIATES LBP IN A MOUSE MODEL. THIS MAY BE MEDIATED, IN PART, BY CHANGES IN THE EPIGENETIC REGULATORY MACHINERY IN DEGENERATING IVDS. EPIGENETIC ALTERATIONS DUE TO A LIFESTYLE CHANGE COULD HAVE A LONG-LASTING THERAPEUTIC IMPACT BY CHANGING TISSUE HOMEOSTASIS IN IVDS. CLINICAL SIGNIFICANCE: THIS STUDY CONFIRMED THE THERAPEUTIC BENEFITS OF EXERCISE ON LBP AND SUGGESTS THAT EXERCISE RESULTS IN SEX-SPECIFIC ALTERATIONS IN EPIGENETIC REGULATION IN IVDS. ELUCIDATING THE EFFECTS OF EXERCISE ON EPIGENETIC REGULATION MAY ENABLE THE DISCOVERY OF NOVEL GENE TARGETS OR NEW STRATEGIES TO IMPROVE THE TREATMENT OF CHRONIC LBP.	2021	

18  141  28 ABERRANT DNA METHYLATION OF MTOR PATHWAY GENES PROMOTES INFLAMMATORY ACTIVATION OF IMMUNE CELLS IN DIABETIC KIDNEY DISEASE. DNA METHYLATION HAS BEEN IMPLICATED IN THE PATHOGENESIS OF DIABETIC KIDNEY DISEASE (DKD), BUT THE UNDERLYING MECHANISMS REMAIN UNCLEAR. IN THIS STUDY, WE TESTED THE HYPOTHESIS THAT ABERRANT DNA METHYLATION IN PERIPHERAL IMMUNE CELLS CONTRIBUTES TO DKD PROGRESSION. WE SHOWED THAT LEVELS OF DNA METHYLTRANSFERASE 1 (DNMT1), A KEY ENZYME FOR DNA METHYLATION, WERE INCREASED ALONG WITH INFLAMMATORY ACTIVITY OF PERIPHERAL BLOOD MONONUCLEAR CELLS IN DKD PATIENTS. INHIBITION OF DNMT1 WITH 5-AZA-2'-DEOXYCYTIDINE (5-AZA) MARKEDLY INCREASED THE PROPORTION OF CD4(+)CD25(+) REGULATORY T CELLS IN PERIPHERAL BLOOD MONONUCLEAR CELLS IN CULTURE AND IN DIABETIC ANIMALS. ADOPTIVE TRANSFER OF IMMUNE CELLS FROM 5-AZA-TREATED ANIMALS SHOWED BENEFICIAL EFFECTS ON THE HOST IMMUNE SYSTEM, RESULTING IN A SIGNIFICANT IMPROVEMENT OF DKD. USING GENOME-WIDE DNA METHYLATION ASSAYS, WE IDENTIFIED THE DIFFERENTIALLY METHYLATED CYTOSINES IN THE PROMOTER REGIONS OF MAMMALIAN TARGET OF RAPAMYCIN (MTOR) REGULATORS IN PERIPHERAL BLOOD MONONUCLEAR CELLS OF DIABETIC PATIENTS. FURTHER, MRNA ARRAYS CONFIRMED THE CONSISTENT INDUCTION OF GENES EXPRESSED IN THE MTOR PATHWAY. IMPORTANTLY, DOWN-REGULATION OF DNMT1 EXPRESSION VIA RNA INTERFERENCE RESULTED IN PROMINENT CYTOSINE DEMETHYLATION OF MTOR NEGATIVE REGULATORS AND SUBSEQUENT DECREASE OF MTOR ACTIVITY. LASTLY, MODULATION OF MTOR RESULTED IN CHANGES IN THE EFFECT OF 5-AZA ON DIABETIC IMMUNE CELLS. THUS, UP-REGULATION OF DNMT1 IN DIABETIC IMMUNE CELLS INDUCES ABERRANT CYTOSINE METHYLATION OF THE UPSTREAM REGULATORS OF MTOR, LEADING TO PATHOGENIC ACTIVATION OF THE MTOR PATHWAY AND CONSEQUENT INFLAMMATION IN DIABETIC KIDNEYS. HENCE, THIS STUDY HIGHLIGHTS THERAPEUTIC POTENTIAL OF TARGETING EPIGENETIC EVENTS IN IMMUNE SYSTEM FOR TREATING DKD.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
19  213  34 ACUTE AND CHRONIC ELECTROCONVULSIVE SEIZURES (ECS) DIFFERENTIALLY REGULATE THE EXPRESSION OF EPIGENETIC MACHINERY IN THE ADULT RAT HIPPOCAMPUS. BACKGROUND: ELECTROCONVULSIVE SEIZURE TREATMENT IS A FAST-ACTING ANTIDEPRESSANT THERAPY THAT EVOKES RAPID TRANSCRIPTIONAL, NEUROGENIC, AND BEHAVIORAL CHANGES. EPIGENETIC MECHANISMS CONTRIBUTE TO ALTERED GENE REGULATION, WHICH UNDERLIES THE NEUROGENIC AND BEHAVIORAL EFFECTS OF ELECTROCONVULSIVE SEIZURE. WE HYPOTHESIZED THAT ELECTROCONVULSIVE SEIZURE MAY MODULATE THE EXPRESSION OF EPIGENETIC MACHINERY, THUS ESTABLISHING POTENTIAL ALTERATIONS IN THE EPIGENETIC LANDSCAPE. METHODS: WE EXAMINED THE INFLUENCE OF ACUTE AND CHRONIC ELECTROCONVULSIVE SEIZURE ON THE GENE EXPRESSION OF HISTONE MODIFIERS, NAMELY HISTONE ACETYLTRANSFERASES, HISTONE DEACETYLASES, HISTONE METHYLTRANSFERASES, AND HISTONE (LYSINE) DEMETHYLASES AS WELL AS DNA MODIFYING ENZYMES, INCLUDING DNA METHYLTRANSFERASES, DNA DEMETHYLASES, AND METHYL-CPG-BINDING PROTEINS IN THE HIPPOCAMPI OF ADULT MALE WISTAR RATS USING QUANTITATIVE REAL TIME-PCR ANALYSIS. FURTHER, WE EXAMINED THE INFLUENCE OF ACUTE AND CHRONIC ELECTROCONVULSIVE SEIZURE ON GLOBAL AND RESIDUE-SPECIFIC HISTONE ACETYLATION AND METHYLATION LEVELS WITHIN THE HIPPOCAMPUS, A BRAIN REGION IMPLICATED IN THE CELLULAR AND BEHAVIORAL EFFECTS OF ELECTROCONVULSIVE SEIZURE. RESULTS: ACUTE AND CHRONIC ELECTROCONVULSIVE SEIZURE INDUCED A PRIMARILY UNIQUE, AND IN CERTAIN CASES BIDIRECTIONAL, REGULATION OF HISTONE AND DNA MODIFIERS, AND METHYL-CPG-BINDING PROTEINS, WITH AN OVERLAPPING PATTERN OF GENE REGULATION RESTRICTED TO SIRT4, MLL3, JMJD3, GADD45B, TET2, AND TET3. GLOBAL HISTONE ACETYLATION AND METHYLATION LEVELS WERE PREDOMINANTLY UNCHANGED, WITH THE EXCEPTION OF A SIGNIFICANT DECLINE IN H3K9 ACETYLATION IN THE HIPPOCAMPUS FOLLOWING CHRONIC ELECTROCONVULSIVE SEIZURE. CONCLUSIONS: ELECTROCONVULSIVE SEIZURE TREATMENT EVOKES THE TRANSCRIPTIONAL REGULATION OF SEVERAL HISTONE AND DNA MODIFIERS, AND METHYL-CPG-BINDING PROTEINS WITHIN THE HIPPOCAMPUS, WITH A PREDOMINANTLY DISTINCT PATTERN OF REGULATION INDUCED BY ACUTE AND CHRONIC ELECTROCONVULSIVE SEIZURE.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
20 2417  33 EPIGENETIC SIGNATURE OF CHRONIC CEREBRAL HYPOPERFUSION AND BENEFICIAL EFFECTS OF S-ADENOSYLMETHIONINE IN RATS. CHRONIC CEREBRAL HYPOPERFUSION IS ASSOCIATED WITH COGNITIVE DECLINE IN AGING AND AGE-RELATED NEURODEGENERATIVE DISEASE. EPIGENETIC MECHANISMS ARE INVOLVED IN THE MAINTENANCE OF LONG-TERM HYPOXIA-ADAPTED CELLULAR PHENOTYPES. IN THE PRESENT STUDY, THE EPIGENETIC SIGNATURES SUCH AS DNA METHYLATION AND HISTONE ACETYLATION, AS WELL AS S-ADENOSYLMETHIONINE (SAM) CYCLE USING CHRONIC CEREBRAL HYPOPERFUSION RAT MODEL WERE EXPLORED. CHRONIC CEREBRAL HYPOXIA-INDUCED GLOBAL DNA HYPERMETHYLATION ASSOCIATED WITH THE INCREASE OF DNA METHYLTRANSFERASE (DNMT) 3A AS WELL AS ALTERATION OF SAM CYCLE. MEANWHILE, AN ENHANCED LEVEL OF GLOBAL HISTONE H4 ACETYLATION ACCOMPANIED WITH THE UPREGULATION OF HISTONE ACETYLTRANSFERASE, P300/CREB-BINDING PROTEIN (CBP), AND THE DOWNREGULATION OF HISTONE DEACETYLASES (HDACS), WAS ALSO OBSERVED. SAM COULD IMPROVE SPATIAL CAPACITY THROUGH THE UPREGULATION OF ACETYLCHOLINE AND BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) RATHER THAN ALTERATION OF DNA METHYLATION LEVELS. IN CONCLUSION, WE HAVE DEMONSTRATED A GENOME-WIDE ADJUSTMENT OF DNA METHYLATION AND HISTONE ACETYLATION UNDER CHRONIC CEREBRAL HYPOXIC CONDITIONS IN A RAT'S BRAIN. THESE EPIGENETIC SIGNATURES MAY REPRESENT AN ADDITIONAL MECHANISM TO PROMOTE AND MAINTAIN A HYPOXIC-ADAPTED CELLULAR RESPONDS WITH A POTENTIAL ROLE IN MEMORY DEFICITS.	2014