1 4989 117 PCSK9 IS INCREASED IN CEREBROSPINAL FLUID OF INDIVIDUALS WITH ALCOHOL USE DISORDER. BACKGROUND: RECENT STUDIES HAVE SHOWN THAT ALCOHOL USE AFFECTS THE REGULATION AND EXPRESSION OF PROPROTEIN CONVERTASE SUBTILISIN/KEXIN 9 (PCSK9). WHILE A MAJOR ROLE OF PCSK9 IN HEPATIC FUNCTION AND LIPID REGULATION HAS BEEN CLEARLY ESTABLISHED, OTHER PLEIOTROPIC EFFECTS REMAIN POORLY UNDERSTOOD. EXISTING RESEARCH SUGGESTS A POSITIVE ASSOCIATION BETWEEN PCSK9 EXPRESSION IN THE BRAIN AND PSYCHOPATHOLOGY, WITH INCREASED LEVELS OF PCSK9 IN THE CEREBROSPINAL FLUID (CSF) OF INDIVIDUALS WITH DEMENTIA AND EPIGENETIC MODIFICATIONS OF PCSK9 ASSOCIATED WITH ALCOHOL USE DISORDER (AUD). IN THIS STUDY, WE HYPOTHESIZED THAT CHRONIC ALCOHOL USE WOULD INCREASE PCSK9 EXPRESSION IN CSF. METHODS: PCSK9 LEVELS IN CSF WERE MEASURED IN INDIVIDUALS WITH AUD (N = 42) ADMITTED TO AN INPATIENT REHABILITATION PROGRAM AND CONTROLS (N = 25). CSF SAMPLES IN AUD WERE ASSESSED AT 2 TIME POINTS, AT DAY 5 AND DAY 21 AFTER ADMISSION. FURTHERMORE, PLASMA SAMPLES WERE COLLECTED AND MEASURED FROM THE INDIVIDUALS WITH AUD. RESULTS: PCSK9 IN CSF WAS SIGNIFICANTLY INCREASED IN THE AUD GROUP AT DAY 5 AND DAY 21 COMPARED TO THE CONTROLS (P < 0.0001). PLASMA PCSK9 LEVELS WERE CORRELATED POSITIVELY WITH CSF PCSK9 LEVELS IN AUD (P = 0.0493). CONCLUSIONS: OUR DATA SUGGEST THAT PCSK9 IS ELEVATED IN THE CSF OF INDIVIDUALS WITH AUD, WHICH MAY INDICATE A POTENTIAL ROLE OF PCSK9 IN AUD. ADDITIONAL STUDIES ARE NECESSARY TO FURTHER ELUCIDATE THE FUNCTIONS OF PCSK9 IN THE BRAIN. 2019 2 4257 41 METHYLOMIC PROFILING AND REPLICATION IMPLICATES DEREGULATION OF PCSK9 IN ALCOHOL USE DISORDER. ALCOHOL USE DISORDER (AUD) IS A COMMON AND CHRONIC DISORDER WITH SUBSTANTIAL EFFECTS ON PERSONAL AND PUBLIC HEALTH. THE UNDERLYING PATHOPHYSIOLOGY IS POORLY UNDERSTOOD BUT STRONG EVIDENCE SUGGESTS SIGNIFICANT ROLES OF BOTH GENETIC AND EPIGENETIC COMPONENTS. GIVEN THAT ALCOHOL AFFECTS MANY ORGAN SYSTEMS, WE PERFORMED A CROSS-TISSUE AND CROSS-PHENOTYPIC ANALYSIS OF GENOME-WIDE METHYLOMIC VARIATION IN AUD USING SAMPLES FROM 3 DISCOVERY, 4 REPLICATION, AND 2 TRANSLATIONAL COHORTS. WE IDENTIFIED A DIFFERENTIALLY METHYLATED REGION IN THE PROMOTER OF THE PROPROTEIN CONVERTASE SUBTILISIN/KEXIN 9 (PCSK9) GENE THAT WAS ASSOCIATED WITH DISEASE PHENOTYPES. BIOLOGICAL VALIDATION SHOWED THAT PCSK9 PROMOTER METHYLATION IS CONSERVED ACROSS TISSUES AND POSITIVELY CORRELATED WITH EXPRESSION. REPLICATION IN AUD DATASETS CONFIRMED PCSK9 HYPOMETHYLATION AND A TRANSLATIONAL MOUSE MODEL OF AUD SHOWED THAT ALCOHOL EXPOSURE LEADS TO PCSK9 DOWNREGULATION. PCSK9 IS PRIMARILY EXPRESSED IN THE LIVER AND REGULATES LOW-DENSITY LIPOPROTEIN CHOLESTEROL (LDL-C). OUR FINDING OF ALCOHOL-INDUCED EPIGENETIC REGULATION OF PCSK9 REPRESENTS ONE OF THE UNDERLYING MECHANISMS BETWEEN THE WELL-KNOWN EFFECTS OF ALCOHOL ON LIPID METABOLISM AND CARDIOVASCULAR RISK, WITH LIGHT ALCOHOL USE GENERALLY BEING PROTECTIVE WHILE CHRONIC HEAVY USE HAS DETRIMENTAL HEALTH OUTCOMES. 2018 3 5285 39 PROPROTEIN CONVERTASE SUBTILISIN/KEXIN TYPE 9 (PCSK9) IN THE BRAIN AND RELEVANCE FOR NEUROPSYCHIATRIC DISORDERS. PROPROTEIN CONVERTASE SUBTILISIN/KEXIN TYPE 9 (PCSK9) HAS LONG BEEN STUDIED IN THE LIVER DUE TO ITS REGULATION OF PLASMA LOW-DENSITY LIPOPROTEIN CHOLESTEROL (LDL-C) AND ITS CAUSAL ROLE IN FAMILIAL HYPERCHOLESTEROLEMIA. ALTHOUGH PCSK9 WAS FIRST DISCOVERED IN CEREBELLAR NEURONS UNDERGOING APOPTOSIS, ITS FUNCTION IN THE CENTRAL NERVOUS SYSTEM (CNS) IS LESS CLEAR. PCSK9 HAS BEEN SHOWN TO BE INVOLVED IN NEURONAL DIFFERENTIATION, LDL RECEPTOR FAMILY METABOLISM, APOPTOSIS, AND INFLAMMATION IN THE BRAIN, BUT IN VITRO AND IN VIVO STUDIES OFFER CONTRADICTORY FINDINGS. PCSK9 EXPRESSION IN THE ADULT BRAIN IS LOW BUT IS HIGHLY UPREGULATED DURING DISEASE STATES. CEREBRAL SPINAL FLUID (CSF) PCSK9 CONCENTRATIONS ARE CORRELATED WITH NEURAL TUBE DEFECTS AND NEURODEGENERATIVE DISEASES IN HUMAN PATIENTS. EPIGENETIC STUDIES REVEAL THAT CHRONIC ALCOHOL USE MAY MODULATE METHYLATION OF THE PCSK9 GENE AND GENETIC STUDIES SHOW THAT PATIENTS WITH GAIN-OF-FUNCTION PCSK9 VARIANTS HAVE HIGHER LDL-C AND AN INCREASED RISK OF ISCHEMIC STROKE. EARLY SAFETY STUDIES OF THE PCSK9 INHIBITORS EVOLOCUMAB AND ALIROCUMAB, USED TO TREAT HYPERCHOLESTEROLEMIA, HINTED THAT PCSK9 INHIBITION MAY NEGATIVELY IMPACT COGNITION BUT MORE RECENT, LONGER-TERM CLINICAL TRIALS FOUND NO ADVERSE NEUROCOGNITIVE EVENTS. THE PURPOSE OF THIS REVIEW IS TO ELUCIDATE THE ROLE OF PCSK9 IN THE BRAIN, PARTICULARLY ITS ROLE IN DISEASE PATHOGENESIS. 2020 4 5682 34 SHORT-TERM CHANGES IN GLOBAL METHYLATION AND HYDROXYMETHYLATION DURING ALCOHOL DETOXIFICATION. ALCOHOL DEPENDENCE IS A COMMON PUBLIC HEALTH PROBLEM AND EPIGENETICS MAY OFFER NEW ASPECTS IN UNDERSTANDING THE BIOLOGICAL AND GENETIC UNDERPINNINGS AND IMPROVE TREATMENT OF THIS COMPLEX DISEASE. SUPPOSEDLY, METHYLATION AND HYDROXYMETHYLATION ARE ALTERED IN BRAIN TISSUES AND IN SYNAPSE-RELATED GENES DUE TO CHRONIC ALCOHOL INTAKE AND DURING WITHDRAWAL. TO ASSESS POTENTIAL EPIGENETIC CHANGES AFTER CESSATION OF CHRONIC ALCOHOL INTAKE, WE COMPARED 23 ALCOHOL-DEPENDENT INDIVIDUALS DURING INPATIENT ALCOHOL DETOXIFICATION WITH 13 CAREFULLY MATCHED CONTROLS. BLOOD SAMPLES WERE TAKEN ON THE DAY OF ADMISSION, AFTER ONE AND AFTER TWO WEEKS AT THE END OF INPATIENT TREATMENT. GENOME-WIDE GLOBAL METHYLATION AND GLOBAL DNA HYDROXYMETHYLATION WERE COMPARED ACROSS GROUPS. THERE WERE SIGNIFICANT DIFFERENCES IN GLOBAL METHYLATION ACROSS TIME FROM ADMISSION TO ONE AND TWO WEEKS OF INPATIENT WITHDRAWAL (P < 0.001). THESE FINDINGS WERE PARALLELED TO CHANGES IN GLOBAL DNA HYDROXYMETHYLATION ACROSS TIME WHEN AGE WAS EMPLOYED AS A COFACTOR (P < 0.001). SEVERAL POTENTIALLY INFLUENCING VARIABLES LIKE SEVERITY OF WITHDRAWAL, DOSE OF WITHDRAWAL MEDICATION AND ALCOHOL INTAKE BEFORE ADMISSION DID NOT YIELD SIGNIFICANT INFLUENCE ON EPIGENETIC CHANGES. THE RESULTS CONFIRM PREVIOUS FINDINGS OF SIGNIFICANT ALTERATIONS OF EPIGENETIC PATTERNS DURING ALCOHOL INTOXICATION AND PRESENT FOR THE FIRST TIME HYDROXYMETHYLATION CHANGES IN THESE INDIVIDUALS. 2019 5 1967 43 EPIGENETIC ALTERATION OF THE DOPAMINE TRANSPORTER GENE IN ALCOHOL-DEPENDENT PATIENTS IS ASSOCIATED WITH AGE. CHRONIC ALCOHOL ABUSE AND DEPENDENCE ARE ASSOCIATED WITH DYSFUNCTIONAL DOPAMINERGIC NEUROTRANSMISSION IN MESOCORTICOLIMBIC CIRCUITS. GENETIC AND ENVIRONMENTAL FACTORS HAVE BEEN SHOWN TO MODULATE SUSCEPTIBILITY TO ALCOHOL DEPENDENCE, AND BOTH MAY ACT THROUGH EPIGENETIC MECHANISMS THAT CAN MODULATE GENE EXPRESSION, E.G. DNA METHYLATION AT CPG SITES. RECENT STUDIES HAVE SUGGESTED THAT DNA METHYLATION PATTERNS MAY CHANGE OVER TIME. HOWEVER, FEW DATA ARE AVAILABLE CONCERNING THE RATE OF THESE CHANGES IN SPECIFIC GENES. A RECENT STUDY FOUND THAT HYPERMETHYLATION OF THE PROMOTER OF THE DOPAMINE TRANSPORTER (DAT) GENE WAS POSITIVELY CORRELATED WITH ALCOHOL DEPENDENCE AND NEGATIVELY CORRELATED WITH ALCOHOL CRAVING. THE AIM OF THE PRESENT STUDY WAS TO REPLICATE THESE FINDINGS IN A LARGER SAMPLE OF ALCOHOL-DEPENDENT PATIENTS AND POPULATION-BASED CONTROLS MATCHED FOR AGE AND SEX. NO DIFFERENCE IN METHYLATION LEVEL WAS OBSERVED BETWEEN PATIENTS AND CONTROLS, AND NO DIFFERENCE IN METHYLATION LEVEL WAS OBSERVED BEFORE AND AFTER ALCOHOL WITHDRAWAL IN PATIENTS. HOWEVER, PATIENTS WITH MORE SEVERE CRAVING SHOWED A TREND TOWARDS LOWER DAT METHYLATION LEVELS (P = 0.07), WHICH IS CONSISTENT WITH PREVIOUS FINDINGS. FURTHERMORE, IN OUR OVERALL SAMPLE, DAT METHYLATION LEVELS INCREASED WITH AGE. INTERESTINGLY, A SEPARATE ANALYSIS OF PATIENTS SUGGESTED THAT THIS FINDING WAS MAINLY DRIVEN BY THE PATIENT GROUP. ALTHOUGH THE PRESENT DATA DO NOT CLARIFY WHETHER CHRONIC ALCOHOL ABUSE IS RESPONSIBLE FOR THIS PHENOMENON OR MERELY ENHANCES AN AGEING-SPECIFIC PROCESS, OUR FINDINGS SUGGEST THAT HYPERMETHYLATION IN ALCOHOL-DEPENDENT PATIENTS IS A CONSEQUENCE, RATHER THAN A CAUSE, OF THE DISORDER. 2014 6 1497 43 DNA METHYLATION AGE IS ACCELERATED IN ALCOHOL DEPENDENCE. ALCOHOL DEPENDENCE (ALC) IS A CHRONIC, RELAPSING DISORDER THAT INCREASES THE BURDEN OF CHRONIC DISEASE AND SIGNIFICANTLY CONTRIBUTES TO NUMEROUS PREMATURE DEATHS EACH YEAR. PREVIOUS RESEARCH SUGGESTS THAT CHRONIC, HEAVY ALCOHOL CONSUMPTION IS ASSOCIATED WITH DIFFERENTIAL DNA METHYLATION PATTERNS. IN ADDITION, DNA METHYLATION LEVELS AT CERTAIN CPG SITES HAVE BEEN CORRELATED WITH AGE. WE USED AN EPIGENETIC CLOCK TO INVESTIGATE THE POTENTIAL ROLE OF EXCESSIVE ALCOHOL CONSUMPTION IN EPIGENETIC AGING. WE EXPLORED THIS QUESTION IN FIVE INDEPENDENT COHORTS, INCLUDING DNA METHYLATION DATA DERIVED FROM DATASETS FROM BLOOD (N = 129, N = 329), LIVER (N = 92, N = 49), AND POSTMORTEM PREFRONTAL CORTEX (N = 46). ONE BLOOD DATASET AND ONE LIVER TISSUE DATASET OF INDIVIDUALS WITH ALC EXHIBITED POSITIVE AGE ACCELERATION (P < 0.0001 AND P = 0.0069, RESPECTIVELY), WHEREAS THE OTHER BLOOD AND LIVER TISSUE DATASETS BOTH EXHIBITED TRENDS OF POSITIVE AGE ACCELERATION THAT WERE NOT SIGNIFICANT (P = 0.83 AND P = 0.57, RESPECTIVELY). PREFRONTAL CORTEX TISSUE EXHIBITED A TREND OF NEGATIVE AGE ACCELERATION (P = 0.19). THESE RESULTS SUGGEST THAT EXCESSIVE ALCOHOL CONSUMPTION MAY BE ASSOCIATED WITH EPIGENETIC AGING IN A TISSUE-SPECIFIC MANNER AND WARRANTS FURTHER INVESTIGATION USING MULTIPLE TISSUE SAMPLES FROM THE SAME INDIVIDUALS. 2018 7 522 42 ASSOCIATIONS BETWEEN NEIGHBORHOOD SOCIOECONOMIC DEPRIVATION, IFNGAMMA, AND HIGH-DENSITY LIPOPROTEIN PARTICLE SIZE: DATA FROM THE WASHINGTON, D.C. CARDIOVASCULAR HEALTH AND NEEDS ASSESSMENT. INTRODUCTION: NEIGHBORHOOD SOCIOECONOMIC DEPRIVATION IS ASSOCIATED WITH INCREASED CARDIOVASCULAR RISK FACTORS, INCLUDING INFLAMMATION. INFLAMMATION PLAYS AN IMPORTANT ROLE IN MODIFYING THE CARDIOPROTECTIVE FUNCTION OF HIGH-DENSITY LIPOPROTEIN (HDL). MOREOVER, RECENT STUDIES SUGGEST THAT VERY HIGH HDL IS ASSOCIATED WITH ADVERSE CARDIOVASCULAR DISEASE (CVD) OUTCOMES. THUS, WE SOUGHT TO EXPLORE THE RELATIONSHIPS BETWEEN NEIGHBORHOOD SOCIOECONOMIC DEPRIVATION AS A MARKER OF CHRONIC STRESS, INFLAMMATION, PROPROTEIN CONVERTASE SUBTILISIN/KEXIN TYPE 9 (PCSK9) (A CORE COMPONENT OF THE HDL PROTEOME), HDL CHARACTERISITCS, AND BIOLOGICAL AGING AS A PREDICTOR OF CVD AND ALL-CAUSE MORTALITY. METHODS: SIXTY AFRICAN AMERICAN SUBJECTS WERE RECRUITED TO THE NIH CLINICAL CENTER AS PART OF A COMMUNITY-BASED PARTICIPATORY RESEARCH-DESIGNED OBSERVATIONAL STUDY. NEIGHBORHOOD DEPRIVATION INDEX (NDI), A MARKER OF NEIGHBORHOOD SOCIOECONOMIC DEPRIVATION, WAS MEASURED USING US CENSUS DATA. HDL CHARACTERISTICS (CHOLESTEROL, PARTICLE NUMBER, SIZE, SUBSPECIES) WERE DETERMINED FROM NMR LIPOPROTEIN PROFILING, AND PLASMA CYTOKINES (IL-1BETA, IL-6, IL-8, TNFALPHA, IFNGAMMA) WERE MEASURED USING AN ELISA-BASED MULTIPLEX TECHNIQUE. EPIGENETIC CLOCK BIOMARKERS OF AGING WERE MEASURED USING DNA METHYLATION DATA OBTAINED FROM PARTICIPANTS' BUFFY COAT SAMPLES. WE USED LINEAR REGRESSION MODELING ADJUSTED FOR ATHEROSCLEROTIC CARDIOVASCULAR DISEASE (ASCVD) RISK SCORE, BODY MASS INDEX (BMI), AND LIPID-LOWERING MEDICATION USE TO INVESTIGATE RELATIONSHIPS OF INTEREST. RESULTS: NDI DIRECTLY ASSOCIATED WITH LARGE HDL PARTICLE COUNT (H7P) AND IFNGAMMA AND TRENDED TOWARD SIGNIFICANCE WITH HDL-C AND PCSK9. IFNGAMMA AND PCSK9 THEN DIRECTLY ASSOCIATED WITH H7P. H7P ALSO DIRECTLY ASSOCIATED WITH HIGHER DNA METHYLATION PHENOTYPIC AGE (PHENOAGE). CONCLUSION: WE HIGHLIGHT ASSOCIATIONS BETWEEN NEIGHBORHOOD SOCIOECONOMIC DEPRIVATION, IFNGAMMA, PCSK9, HDL SUBSPECIES, AND EPIGENETIC BIOMARKERS OF AGING. TAKEN TOGETHER, OUR FINDINGS SUGGEST INDIRECT PATHWAYS LINKING NEIGHBORHOOD DEPRIVATION-RELATED STRESS AND INFLAMMATION TO HDL AND IMMUNE EPIGENETIC CHANGES. MOREOVER, THESE RESULTS ADD TO RECENT WORK SHOWING THE PATHOGENICITY OF HIGH HDL LEVELS AND UNDERSCORE THE NEED TO UNDERSTAND HOW CHRONIC STRESS-RELATED INFLAMMATION AND LIPOPROTEIN SUBSPECIES RELATE TO CVD RISK ACROSS DIVERSE POPULATIONS. 2023 8 6311 31 THE RELATION BETWEEN DNA METHYLATION PATTERNS AND SERUM CYTOKINE LEVELS IN COMMUNITY-DWELLING ADULTS: A PRELIMINARY STUDY. BACKGROUND: THE LEVELS OF CIRCULATING CYTOKINES FLUCTUATE WITH AGE, ACUTE ILLNESS, AND CHRONIC DISEASE, AND ARE PREDICTIVE OF MORTALITY; THIS IS ALSO TRUE FOR PATTERNS OF DNA (CPG) METHYLATION. GIVEN THAT IMMUNE CELLS ARE PARTICULARLY SENSITIVE TO CHANGES IN THE CONCENTRATION OF CYTOKINES IN THEIR MICROENVIRONMENT, WE HYPOTHESIZED THAT SERUM LEVELS OF TNF, IL-6, IL-8 AND IL-10 WOULD CORRELATE WITH GENOME-WIDE ALTERATIONS IN THE DNA METHYLATION LEVELS OF BLOOD LEUKOCYTES. TO TEST THIS, WE EVALUATED COMMUNITY-DWELLING ADULTS (N = 14; 48-78 YEARS OLD) RECRUITED TO A PILOT STUDY FOR THE CANADIAN LONGITUDINAL STUDY ON AGING (CLSA), EXAMINING DNA METHYLATION PATTERNS IN PERIPHERAL BLOOD MONONUCLEAR CELLS USING THE ILLUMINA HUMANMETHYLATION 450 K BEADCHIP. RESULTS: WE SHOW THAT, APART FROM AGE, SERUM IL-10 LEVELS EXHIBITED THE MOST SUBSTANTIAL ASSOCIATION TO DNA METHYLATION PATTERNS, FOLLOWED BY TNF, IL-6 AND IL-8. FURTHERMORE, WHILE THE LEVELS OF THESE CYTOKINES WERE HIGHER IN ELDERLY ADULTS, NO ASSOCIATIONS WITH EPIGENETIC ACCELERATED AGING, DERIVED USING THE EPIGENETIC CLOCK, WERE OBSERVED. CONCLUSIONS: AS A PRELIMINARY STUDY WITH A SMALL SAMPLE SIZE, THE CONCLUSIONS DRAWN FROM THIS WORK MUST BE VIEWED WITH CAUTION; HOWEVER, OUR OBSERVATIONS ARE ENCOURAGING AND CERTAINLY WARRANT MORE SUITABLY POWERED STUDIES OF THIS RELATIONSHIP. 2017 9 2625 38 EPIGENOME-WIDE ASSOCIATION STUDY AND MULTI-TISSUE REPLICATION OF INDIVIDUALS WITH ALCOHOL USE DISORDER: EVIDENCE FOR ABNORMAL GLUCOCORTICOID SIGNALING PATHWAY GENE REGULATION. ALCOHOL USE DISORDER (AUD) IS A CHRONIC DEBILITATING DISORDER WITH LIMITED TREATMENT OPTIONS AND POORLY DEFINED PATHOPHYSIOLOGY. THERE ARE SUBSTANTIAL GENETIC AND EPIGENETIC COMPONENTS; HOWEVER, THE UNDERLYING MECHANISMS CONTRIBUTING TO AUD REMAIN LARGELY UNKNOWN. WE CONDUCTED THE LARGEST DNA METHYLATION EPIGENOME-WIDE ASSOCIATION STUDY (EWAS) ANALYSES CURRENTLY AVAILABLE FOR AUD (TOTAL N = 625) AND EMPLOYED A TOP HIT REPLICATION (N = 4798) USING A CROSS-TISSUE/CROSS-PHENOTYPIC APPROACH WITH THE GOAL OF IDENTIFYING NOVEL EPIGENETIC TARGETS RELEVANT TO AUD. RESULTS SHOW THAT A NETWORK OF DIFFERENTIALLY METHYLATED REGIONS IN GLUCOCORTICOID SIGNALING AND INFLAMMATION-RELATED GENES WERE ASSOCIATED WITH ALCOHOL USE BEHAVIORS. A TOP PROBE CONSISTENTLY ASSOCIATED ACROSS ALL COHORTS WAS LOCATED IN THE LONG NON-CODING RNA GROWTH ARREST SPECIFIC FIVE GENE (GAS5) (P < 10(-24)). GAS5 HAS BEEN IMPLICATED IN REGULATING TRANSCRIPTIONAL ACTIVITY OF THE GLUCOCORTICOID RECEPTOR AND HAS MULTIPLE FUNCTIONS RELATED TO APOPTOSIS, IMMUNE FUNCTION AND VARIOUS CANCERS. ENDOPHENOTYPIC ANALYSES USING PERIPHERAL CORTISOL LEVELS AND NEUROIMAGING PARADIGMS SHOWED THAT METHYLOMIC VARIATION IN GAS5 NETWORK-RELATED PROBES WERE ASSOCIATED WITH STRESS PHENOTYPES. POSTMORTEM BRAIN ANALYSES DOCUMENTED INCREASED GAS5 EXPRESSION IN THE AMYGDALA OF INDIVIDUALS WITH AUD. OUR DATA SUGGEST THAT ALCOHOL USE IS ASSOCIATED WITH DIFFERENTIAL METHYLATION IN THE GLUCOCORTICOID SYSTEM THAT MIGHT INFLUENCE STRESS AND INFLAMMATORY REACTIVITY AND SUBSEQUENTLY RISK FOR AUD. 2021 10 344 36 ALTERED BDNF METHYLATION IN PATIENTS WITH CHRONIC MUSCULOSKELETAL PAIN AND HIGH BIOPSYCHOSOCIAL COMPLEXITY. PURPOSE: THE INTERMED INSTRUMENT, WHICH WAS DEVELOPED TO MEASURE PATIENT'S BIOPSYCHOSOCIAL (BPS) COMPLEXITY, REPRESENTS A POWERFUL DIAGNOSTIC AND THERAPEUTIC TOOL. EPIGENETIC CHANGES ARE THE INTERFACE BETWEEN SIGNALS FROM THE ENVIRONMENT AND GENETIC MODIFICATIONS, AFFECTING GENE EXPRESSION, IN PARTICULAR, BY DNA METHYLATION OF CPG DINUCLEOTIDES IN PROMOTOR REGIONS OF THE CORRESPONDING GENES. THE BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) GENE PLAYS A CRUCIAL ROLE IN THE CENTRAL SENSITIZATION (CS) OF PAIN. IN THIS STUDY, WE HYPOTHESIZED THAT CHRONIC PAIN MODIFIES THE METHYLATION LEVELS OF THE BDNF GENE IN A MANNER THAT IS INTERCONNECTED WITH THE BPS STATUS. PATIENTS AND METHODS: FIFTY-EIGHT CHRONIC MUSCULOSKELETAL PAIN PATIENTS (CMSP) WERE ENROLLED IN THE STUDY. DNA WAS EXTRACTED FROM BLOOD SAMPLES, THE METHYLATION LEVELS OF 13 CPG SITES IN THE BDNF PROMOTER WERE MEASURED BY PYROSEQUENCING, AND ASSOCIATION STUDIES WITH VARIOUS PATIENT PARAMETERS AND THE INTERMED SCORES WERE PERFORMED. RESULTS: INTERESTINGLY, A NEGATIVE CORRELATION (-0.40) WAS FOUND BETWEEN THE TOTAL INTERMED SCORES AND THE AVERAGE CPG METHYLATION VALUES OF THE BDNF GENE, BUT NO CORRELATION WAS OBSERVED WITH THE SEVERITY OF PAIN, DEGREE OF ANXIETY, DEPRESSION, OR KINESIOPHOBIA AND CATASTROPHISM. MOREOVER, THE ASSOCIATION WAS INDEPENDENT OF AGE, SEX AND LEVEL OF COMORBIDITIES. CONCLUSION: THIS RESULT SHOWS THAT CMSP, IN ASSOCIATION WITH ITS BIOPSYCHOSOCIAL CONTEXT, EPIGENETICALLY DECREASES THE DEGREE OF METHYLATION OF THE BDNF PROMOTER AND SHOULD THEREFORE INCREASE THE LEVEL OF BDNF TRANSCRIPTION. IT ALSO SUGGESTS A ROLE OF THE INTERMED TOOL TO DETECT A RELATIONSHIP BETWEEN THE BPS COMPLEXITY AND THE EPIGENETIC CONTROL OF A TARGET GENE. THE POSSIBLE UPREGULATION OF BDNF EXPRESSION MIGHT BE, AT LEAST IN PART, THE SIGNAL FOR CHRONIC PAIN-INDUCED CENTRAL SENSITIZATION (CS). THIS COULD PARTLY EXPLAIN WHY PATIENTS WITH A HIGHER LEVEL OF COMPLEXITY FEEL MORE PAIN THAN THOSE WITH LOWER COMPLEXITY. 2020 11 5498 32 REVIEW: DNA METHYLATION AND ALCOHOL USE DISORDERS: PROGRESS AND CHALLENGES. BACKGROUND AND OBJECTIVES: RISK FOR ALCOHOL USE DISORDERS (AUDS) IS INFLUENCED BY GENE-ENVIRONMENT INTERACTIONS. ENVIRONMENTAL FACTORS CAN AFFECT GENE EXPRESSION THROUGH EPIGENETIC MECHANISMS SUCH AS DNA METHYLATION. THIS REVIEW OUTLINES THE FINDINGS REGARDING THE ASSOCIATION OF DNA METHYLATION AND AUDS. METHODS: WE SEARCHED PUBMED (BY APRIL 2016) AND IDENTIFIED 29 STUDIES THAT EXAMINED THE ASSOCIATION OF DNA METHYLATION AND AUDS. WE ALSO EVALUATED THE METHODS USED IN THESE STUDIES. RESULTS: TWO STUDIES DEMONSTRATED ELEVATED GLOBAL (REPETITIVE ELEMENT) DNA METHYLATION LEVELS IN AUD SUBJECTS. FIFTEEN CANDIDATE GENE STUDIES SHOWED HYPERMETHYLATION OF PROMOTER REGIONS OF SIX GENES (AVP, DNMT3B, HERP, HTR3A, OPRM1, AND SNCA) OR HYPOMETHYLATION OF THE GDAP1 PROMOTER REGION IN AUD SUBJECTS. FIVE GENOME-WIDE DNA METHYLATION STUDIES DEMONSTRATED WIDESPREAD DNA METHYLATION CHANGES ACROSS THE GENOME IN AUD SUBJECTS. SIX STUDIES SHOWED SIGNIFICANT CORRELATIONS OF DNA METHYLATION WITH GENE EXPRESSION IN AUD SUBJECTS. THREE STUDIES REVEALED INTERACTIVE EFFECTS OF GENETIC VARIATION AND DNA METHYLATION ON SUSCEPTIBILITY TO AUDS. MOST STUDIES ANALYZED AUD-ASSOCIATED DNA METHYLATION CHANGES IN THE PERIPHERAL BLOOD; A FEW STUDIES EXAMINED DNA METHYLATION CHANGES IN POSTMORTEM BRAINS OF AUD SUBJECTS. DISCUSSION AND CONCLUSIONS: CHRONIC ALCOHOL CONSUMPTION MAY RESULT IN DNA METHYLATION CHANGES, LEADING TO NEUROADAPTATIONS THAT MAY UNDERLIE SOME OF THE MECHANISMS OF AUD RISK AND PERSISTENCE. FUTURE STUDIES ARE NEEDED TO CONFIRM THE FEW EXISTING RESULTS, AND THEN TO ELUCIDATE WHETHER DNA METHYLATION CHANGES ARE THE CAUSE OR CONSEQUENCE OF AUDS. SCIENTIFIC SIGNIFICANCE: DNA METHYLATION PROFILES MAY BE USED TO ASSESS AUD STATUS OR MONITOR AUD TREATMENT RESPONSE. (AM J ADDICT 2017;26:502-515). 2017 12 6418 40 THE TEMPORAL EXPRESSION OF CIRCULATING MICRORNAS AFTER ACUTE EXPERIMENTAL PAIN IN HUMANS. BACKGROUND: MICRORNAS (MIRNAS) CAN MODULATE SEVERAL BIOLOGICAL SYSTEMS, INCLUDING THE PAIN SYSTEM. THIS STUDY AIMED TO EVALUATE THE TEMPORAL EXPRESSION OF CIRCULATING MIRNAS IN THE PLASMA OF HEALTHY VOLUNTEERS AS A MARKER FOR EPIGENETIC CHANGES BEFORE AND AFTER AN ACUTE, EXPERIMENTAL, PAIN PROVOCATION BY INTRAMUSCULAR HYPERTONIC SALINE INJECTION. METHODS: TWENTY VOLUNTEERS WERE RANDOMLY ALLOCATED INTO TWO GROUPS AND RECEIVED EITHER HYPERTONIC (PAIN) OR ISOTONIC (CONTROL) SALINE INJECTION IN THE FIRST DORSAL INTEROSSEOUS MUSCLE OF THEIR DOMINANT HAND. PAIN INTENSITY WAS CONTINUOUSLY RECORDED FOR 20 MINUTES AFTER INJECTION ON A VAS SCALE FROM 0 TO 100 (0 INDICATES NO PAIN AND 100 THE WORST IMAGINABLE PAIN). BLOOD SAMPLES WERE TAKEN AT BASELINE, 30 MINUTES, 3 HOURS, AND 24 HOURS POST-INJECTION, AND PLASMA WAS SEPARATED. MIRNA EXTRACTS WERE USED FOR RNA SEQUENCING WITH THE ILLUMINA NEXTSEQ PLATFORM. MIRNA TRANSCRIPTS WERE COMPARED BETWEEN THE PAIN AND THE NO-PAIN, CONTROL GROUP AT EVERY TIME POINT. SIGNIFICANT DIFFERENCES WERE CONSIDERED WHEN FOLDS WERE >2 AND THE FALSE DISCOVERY RATE WAS P < 0.05. RESULTS: AFTER 30 MINUTES, 4 MIRNAS WERE SIGNIFICANTLY ALTERED IN THE PAIN GROUP COMPARED TO CONTROLS, WHICH INCREASED TO 24 AFTER 3 HOURS AND TO 42 AFTER 24 HOURS FROM BASELINE (P < 0.0001). TWO MIRNAS WERE CONSISTENTLY UPREGULATED THROUGHOUT THE EXPERIMENT. ENRICHMENT ANALYSIS SHOWED SIGNIFICANT MIRNAS INVOLVED IN BRAIN PERCEPTION OF PAIN, BRAIN SIGNALLING AND RESPONSE TO STIMULI. CONCLUSIONS: THIS EXPLORATORY STUDY IS THE FIRST TO REPORT ON THE TEMPORAL EXPRESSION OF CIRCULATING MIRNAS AFTER AN ACUTE, HUMAN EXPERIMENTAL MUSCLE PAIN MODEL. SIGNIFICANCE: THIS EXPLORATORY STUDY EVALUATED THE TEMPORAL PROFILE OF CIRCULATING MIRNAS IN THE PLASMA OF HEALTHY SUBJECTS AFTER ACUTE EXPERIMENTAL PAIN. SEVERAL MIRNAS WERE ALTERED IN SUBJECTS AT THE TIMES OF FOLLOW-UP AFTER THE ACUTE PAIN MODEL WHEN COMPARED TO CONTROLS. MIRNAS PREVIOUSLY ASSOCIATED WITH PAIN PROCESSES WERE ALTERED IN THE PAIN GROUP. OUR RESULTS, BY SHOWING THE FAST AND PROLONGED MODIFICATIONS OF MIRNA ELICITED BY THE ACUTE EXPERIMENTAL PAIN MODEL, ADD NEW PERSPECTIVES TO THE TOPIC OF EPIGENETICS AND PAIN. 2023 13 314 32 ALCOHOL USE DISORDER IS ASSOCIATED WITH UPREGULATION OF MICRORNA-34A AND MICRORNA-34C IN HIPPOCAMPAL POSTMORTEM TISSUE. BACKGROUND: TO INVESTIGATE EPIGENETIC MECHANISMS POTENTIALLY INVOLVED IN THE COGNITIVE DECLINE ASSOCIATED WITH CHRONIC ALCOHOL INTAKE, WE EVALUATED THE EXPRESSIONS OF THREE MICRO-RNAS (MIR-34A, -34B, AND -34C) HIGHLY EXPRESSED IN THE HIPPOCAMPUS AND INVOLVED IN NEURONAL PHYSIOLOGY AND PATHOLOGY. MIR-34A PARTICIPATES IN FUNCTIONING AND SURVIVAL OF MATURE NEURONS; MIR-34B IS ASSOCIATED WITH ALZHEIMER-LIKE DISORDERS; AND MIR-34C IS IMPLICATED IN THE MEMORY IMPAIRMENT OF ALZHEIMER DISEASE IN RODENTS AND HUMANS. METHODS: A TOTAL OF 69 CASES WERE SELECTED FROM THE BIOBANK FOR AGING STUDIES AND CATEGORIZED ACCORDING TO THE ABSENCE (N = 50) OR PRESENCE (N = 19) OF ALCOHOL USE DISORDER (AUD). CASES PRESENTING WITH NEUROPATHOLOGICAL DIAGNOSES OF DEMENTIAS WERE EXCLUDED. TOTAL RNA WAS EXTRACTED FROM HIPPOCAMPAL PARAFFINIZED SLICES, COMPLEMENTARY DNA WAS SYNTHESIZED FROM MIRS, AND RT-QPCR WAS PERFORMED WITH TAQMAN((R)) ASSAYS. RESULTS: HIGHER EXPRESSIONS OF MIR-34A AND MIR-34C, BUT NOT OF MIR-34B, WERE FOUND IN THE GROUP WITH AUD IN COMPARISON WITH THE GROUP WITHOUT AUD AFTER ADJUSTMENT FOR POTENTIAL CONFOUNDERS (AGE, SEX, BODY MASS INDEX, PRESENCE OF HYPERTENSION, DIABETES MELLITUS, SMOKING, AND PHYSICAL INACTIVITY). CONCLUSIONS: HIPPOCAMPAL UPREGULATION OF MIR-34A AND MIR-34C MAY BE INVOLVED IN THE COGNITIVE DECLINE ASSOCIATED WITH CHRONIC ALCOHOL CONSUMPTION. 2021 14 5034 36 PHARMACOEPIGENETICS OF THE ROLE OF DNA METHYLATION IN MU-OPIOID RECEPTOR EXPRESSION IN DIFFERENT HUMAN BRAIN REGIONS. AIM: EXPOSURE TO OPIOIDS HAS BEEN ASSOCIATED WITH EPIGENETIC EFFECTS. STUDIES IN RODENTS SUGGESTED A ROLE OF VARYING DEGREES OF DNA METHYLATION IN THE DIFFERENTIAL REGULATION OF MU-OPIOID RECEPTOR EXPRESSION ACROSS THE BRAIN. METHODS: IN A TRANSLATIONAL INVESTIGATION, USING TISSUE ACQUIRED POSTMORTEM FROM 21 BRAIN REGIONS OF FORMER OPIATE ADDICTS, REPRESENTING A HUMAN COHORT WITH CHRONIC OPIOID EXPOSURE, MU-OPIOID RECEPTOR EXPRESSION WAS ANALYZED AT THE LEVEL OF DNA METHYLATION, MRNA AND PROTEIN. RESULTS & CONCLUSION: WHILE HIGH OR LOW MU-OPIOID RECEPTOR EXPRESSION SIGNIFICANTLY CORRELATED WITH LOCAL OPRM1 MRNA LEVELS, THERE WAS NO CORRESPONDING ASSOCIATION WITH OPRM1 METHYLATION STATUS. ADDITIONAL EXPERIMENTS IN HUMAN CELL LINES SHOWED THAT CHANGES IN DNA METHYLATION ASSOCIATED WITH CHANGES IN MU-OPIOID EXPRESSION WERE AN ORDER OF MAGNITUDE GREATER THAN DIFFERENCES IN BRAIN. HENCE, DIFFERENT DEGREES OF DNA METHYLATION ASSOCIATED WITH CHRONIC OPIOID EXPOSURE ARE UNLIKELY TO EXERT A MAJOR ROLE IN THE REGION-SPECIFICITY OF MU-OPIOID RECEPTOR EXPRESSION IN THE HUMAN BRAIN. 2016 15 524 35 ASSOCIATIONS OF BDNF GENOTYPE AND PROMOTER METHYLATION WITH ACUTE AND LONG-TERM STROKE OUTCOMES IN AN EAST ASIAN COHORT. BACKGROUND: BRAIN DERIVED NEUROTROPHIC FACTOR (BDNF) HAS BEEN SHOWN TO PLAY AN IMPORTANT ROLE IN POSTSTROKE RECOVERY. BDNF SECRETION IS INFLUENCED BY GENETIC AND EPIGENETIC PROFILES. THIS STUDY AIMED TO INVESTIGATE WHETHER BDNF VAL66MET POLYMORPHISM AND PROMOTER METHYLATION STATUS WERE ASSOCIATED WITH OUTCOMES AT TWO WEEKS AND ONE YEAR AFTER STROKE. METHODS AND FINDINGS: A TOTAL OF 286 PATIENTS WERE EVALUATED AT THE TIME OF ADMISSION AND TWO WEEKS AFTER STROKE, AND 222 (78%) WERE FOLLOWED ONE YEAR LATER IN ORDER TO EVALUATE CONSEQUENCES OF STROKE AT BOTH ACUTE AND CHRONIC STAGES. STROKE OUTCOMES WERE DICHOTOMISED INTO GOOD AND POOR BY THE MODIFIED RANKIN SCALE. STROKE SEVERITY (NATIONAL INSTITUTES OF HEALTH STROKE SCALE), PHYSICAL DISABILITY (BARTHEL INDEX), AND COGNITIVE FUNCTION (MINI-MENTAL STATE EXAMINATION) WERE MEASURED. ASSOCIATIONS OF BDNF GENOTYPE AND METHYLATION STATUS ON STROKE OUTCOMES AND ASSESSMENT SCALE SCORES WERE INVESTIGATED USING LOGISTIC REGRESSION, REPEATED MEASURES ANOVA AND PARTIAL CORRELATION TESTS. BDNF VAL66MET POLYMORPHISM WAS INDEPENDENTLY ASSOCIATED WITH POOR OUTCOME AT 2 WEEKS AND AT 1 YEAR, AND WITH WORSENING PHYSICAL DISABILITY AND COGNITIVE FUNCTION OVER THAT PERIOD. HIGHER BDNF PROMOTER METHYLATION STATUS WAS INDEPENDENTLY ASSOCIATED WITH WORSE OUTCOMES AT 1 YEAR, AND WITH THE WORSENING OF PHYSICAL DISABILITY AND COGNITIVE FUNCTION. NO SIGNIFICANT GENOTYPE-METHYLATION INTERACTIONS WERE FOUND. CONCLUSIONS: A ROLE FOR BDNF IN POSTSTROKE RECOVERY WAS SUPPORTED, AND CLINICAL UTILITY OF BDNF GENETIC AND EPIGENETIC PROFILE AS PROGNOSTIC BIOMARKERS AND A TARGET FOR DRUG DEVELOPMENT WAS SUGGESTED. 2012 16 3080 36 GENOME-WIDE METHYLATION IN ALCOHOL USE DISORDER SUBJECTS: IMPLICATIONS FOR AN EPIGENETIC REGULATION OF THE CORTICO-LIMBIC GLUCOCORTICOID RECEPTORS (NR3C1). ENVIRONMENTAL FACTORS, INCLUDING SUBSTANCE ABUSE AND STRESS, CAUSE LONG-LASTING CHANGES IN THE REGULATION OF GENE EXPRESSION IN THE BRAIN VIA EPIGENETIC MECHANISMS, SUCH AS DNA METHYLATION. WE EXAMINED GENOME-WIDE DNA METHYLATION PATTERNS IN THE PREFRONTAL CORTEX (PFC, BA10) OF 25 PAIRS OF CONTROL AND INDIVIDUALS WITH ALCOHOL USE DISORDER (AUD), USING THE INFINIUM((R)) METHYLATIONEPIC BEADCHIP. WE IDENTIFIED 5254 DIFFERENTIALLY METHYLATED CPGS (P(NOMINAL) < 0.005). BIOINFORMATIC ANALYSES HIGHLIGHTED BIOLOGICAL PROCESSES CONTAINING GENES RELATED TO STRESS ADAPTATION, INCLUDING THE GLUCOCORTICOID RECEPTOR (ENCODED BY NR3C1). CONSIDERING THAT ALCOHOL IS A STRESSOR, WE FOCUSED OUR ATTENTION ON DIFFERENTIALLY METHYLATED REGIONS OF THE NR3C1 GENE AND VALIDATED THE DIFFERENTIAL METHYLATION OF SEVERAL GENES IN THE NR3C1 NETWORK. CHRONIC ALCOHOL DRINKING RESULTS IN A SIGNIFICANT INCREASED METHYLATION OF THE NR3C1 EXON VARIANT 1(H), WITH A PARTICULAR INCREASE IN THE LEVELS OF 5-HYDROXYMETHYLCYTOSINE OVER 5-METHYLCYTOSINE. THESE CHANGES IN DNA METHYLATION WERE ASSOCIATED WITH REDUCED NR3C1 MRNA AND PROTEIN EXPRESSION LEVELS IN PFC, AS WELL AS OTHER CORTICO-LIMBIC REGIONS OF AUD SUBJECTS WHEN COMPARED WITH CONTROLS. FURTHERMORE, WE SHOW THAT THE EXPRESSION OF SEVERAL STRESS-RESPONSIVE GENES (E.G., CRF, POMC, AND FKBP5) IS ALTERED IN THE PFC OF AUD SUBJECTS. THESE STRESS-RESPONSE GENES WERE ALSO CHANGED IN THE HIPPOCAMPUS, A REGION THAT IS HIGHLY SUSCEPTIBLE TO STRESS. THESE DATA SUGGEST THAT ALCOHOL-DEPENDENT ABERRANT DNA METHYLATION OF NR3C1 AND CONSEQUENT CHANGES IN OTHER STRESS-RELATED GENES MIGHT BE FUNDAMENTAL IN THE PATHOPHYSIOLOGY OF AUD AND LAY THE GROUNDWORK FOR TREATMENTS TARGETING THE EPIGENETIC MECHANISMS REGULATING NR3C1 IN AUD. 2021 17 3387 24 HOMOCYSTEINE ASSOCIATED GENOMIC DNA HYPERMETHYLATION IN PATIENTS WITH CHRONIC ALCOHOLISM. HIGHER PLASMA HOMOCYSTEINE CONCENTRATIONS CAN INFLUENCE GENOMIC DNA METHYLATION IN PERIPHERAL BLOOD CELLS. IN THE PRESENT CONTROLLED STUDY WE OBSERVED A SIGNIFICANT INCREASE (10%) OF GENOMIC DNA METHYLATION IN PATIENTS WITH ALCOHOLISM (T = -3.16, DF = 158, P = 0.002) WHICH WAS SIGNIFICANTLY ASSOCIATED WITH THEIR ELEVATED HOMOCYSTEINE LEVELS (MULTIPLE LINEAR REGRESSION, P < 0.001). SINCE METHYLATION OF DNA IS AN IMPORTANT EPIGENETIC FACTOR IN REGULATION OF GENE EXPRESSION THESE FINDINGS MAY HAVE IMPORTANT IMPLICATIONS FOR A POSSIBLE SUBSEQUENT DERANGEMENT OF EPIGENETIC CONTROL THESE PATIENTS. 2004 18 286 38 AGING AND ALCOHOL INTERACT TO ALTER HEPATIC DNA HYDROXYMETHYLATION. BACKGROUND: AGING AND CHRONIC ALCOHOL CONSUMPTION ARE BOTH MODIFIERS OF DNA METHYLATION, BUT IT IS NOT YET KNOWN WHETHER CHRONIC ALCOHOL CONSUMPTION ALSO ALTERS DNA HYDROXYMETHYLATION, A NEWLY DISCOVERED EPIGENETIC MARK PRODUCED BY OXIDATION OF METHYLCYTOSINE. FURTHERMORE, IT HAS NOT BEEN TESTED WHETHER AGING AND ALCOHOL INTERACT TO MODIFY THIS EPIGENETIC PHENOMENON, THEREBY HAVING AN INDEPENDENT EFFECT ON GENE EXPRESSION. METHODS: OLD (18 MONTHS) AND YOUNG (4 MONTHS) MALE C57BL/6 MICE WERE PAIR-FED EITHER A LIEBER-DECARLI LIQUID DIET WITH ALCOHOL (18% OF ENERGY) OR AN ISOCALORIC LIEBER-DECARLI CONTROL DIET FOR 5 WEEKS. GLOBAL DNA HYDROXYMETHYLATION AND DNA METHYLATION WERE ANALYZED FROM HEPATIC DNA USING A NEW LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY METHOD. HEPATIC MRNA EXPRESSION OF THE TET ENZYMES WERE MEASURED VIA QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION. RESULTS: IN YOUNG MICE, MILD CHRONIC ALCOHOL EXPOSURE SIGNIFICANTLY REDUCED GLOBAL DNA HYDROXYMETHYLATION COMPARED WITH CONTROL MICE (0.22 +/- 0.01 VS. 0.29 +/- 0.06%, P = 0.004). ALCOHOL DID NOT SIGNIFICANTLY ALTER HYDROXYMETHYLCYTOSINE LEVELS IN OLD MICE. OLD MICE FED THE CONTROL DIET SHOWED DECREASED GLOBAL DNA HYDROXYMETHYLATION COMPARED WITH YOUNG MICE FED THE CONTROL DIET (0.24 +/- 0.02 VS. 0.29 +/- 0.06%, P = 0.04). THIS MODEL SUGGESTS AN INTERACTION BETWEEN AGING AND ALCOHOL IN DETERMINING DNA HYDROXYMETHYLATION (PINTERACTION = 0.009). EXPRESSION OF TET2 AND TET3 WAS DECREASED IN THE OLD MICE RELATIVE TO THE YOUNG (P < 0.005). CONCLUSIONS: THE OBSERVATION THAT ALCOHOL ALTERS DNA HYDROXYMETHYLATION INDICATES A NEW EPIGENETIC EFFECT OF ALCOHOL. THIS IS THE FIRST STUDY DEMONSTRATING THE INTERACTIVE EFFECTS OF CHRONIC ALCOHOL CONSUMPTION AND AGING ON DNA HYDROXYMETHYLATION. 2014 19 3136 36 GLOBAL DNA METHYLATION LEVELS IN WHITE BLOOD CELLS OF PATIENTS WITH CHRONIC HEROIN USE DISORDER. A PROSPECTIVE STUDY. BACKGROUND: INCREASING SCIENTIFIC EVIDENCE SHOWS THE SIGNIFICANT ROLE OF EPIGENETIC MECHANISMS IN DRUG USE DISORDER, ABSTINENCE AND RELAPSE. STUDIES ON HUMAN SUBJECTS ARE LIMITED COMPARED TO THOSE ON ANIMALS, FOR VARIOUS REASONS SUCH AS POLY-SUBSTANCE ABUSE, HIGH DROP-OUT RATE AND TECHNICAL DIFFICULTIES. OBJECTIVES: OUR GOAL WAS TO EVALUATE WHETHER A MONITORED ABSTINENCE PERIOD OF 21 DAYS COULD INDUCE CHANGES IN GLOBAL DNA METHYLATION IN CHRONIC HEROIN USERS. METHOD: IN THE CURRENT STUDY, WE PRESENT DATA ON GLOBAL DNA METHYLATION ON A SET OF 18 MALE PATIENTS WITH CHRONIC HEROIN USE DISORDER, CAREFULLY SELECTED BASED ON INCLUSION AND EXCLUSION CRITERIA, WHO WERE HOSPITALIZED AND CLOSELY MONITORED DURING A 21-DAY DETOXIFICATION PROGRAM, ONE OF THE FEW WHERE NO OPIOID AGONIST IS ADMINISTERED. THE PARTICIPANTS WERE SAMPLED TWICE, ONCE UPON ENROLMENT TO THE PROGRAM AND ONCE UPON COMPLETION. RESULTS: ACCORDING TO OUR RESULTS, NO DIFFERENCE IN GLOBAL DNA METHYLATION WAS DETECTED BETWEEN SAMPLES COLLECTED UPON ENROLMENT AND SAMPLES COLLECTED UPON COMPLETION OF THE PROGRAM. CONCLUSION: THE FINDINGS OF THIS STUDY DO NOT RULE OUT THE POSSIBILITY THAT THE 21-DAY ABSTINENCE PERIOD WAS NOT LONG ENOUGH TO OBSERVE CHANGES IN GLOBAL DNA METHYLATION, OR THAT ABSTINENCE INDUCED SITE-SPECIFIC METHYLATION CHANGES (BUT NOT GLOBAL CHANGES), THAT CERTAINLY MERIT FURTHER EVALUATION. 2021 20 990 31 CHRONIC SOCIAL STRESS INDUCES DNA METHYLATION CHANGES AT AN EVOLUTIONARY CONSERVED INTERGENIC REGION IN CHROMOSOME X. CHRONIC STRESS RESULTING FROM PROLONGED EXPOSURE TO NEGATIVE LIFE EVENTS INCREASES THE RISK OF MOOD AND ANXIETY DISORDERS. ALTHOUGH CHRONIC STRESS CAN CHANGE GENE EXPRESSION RELEVANT FOR BEHAVIOR, MOLECULAR REGULATORS OF THIS CHANGE HAVE NOT BEEN FULLY DETERMINED. ONE PROCESS THAT COULD PLAY A ROLE IS DNA METHYLATION, AN EPIGENETIC PROCESS WHEREBY A METHYL GROUP IS ADDED ONTO NUCLEOTIDES, PREDOMINANTLY CYTOSINE IN THE CPG CONTEXT, AND WHICH CAN BE INDUCED BY CHRONIC STRESS. IT IS UNKNOWN TO WHAT EXTENT CHRONIC SOCIAL DEFEAT, A MODEL OF HUMAN SOCIAL STRESS, INFLUENCES DNA METHYLATION PATTERNS ACROSS THE GENOME. OUR STUDY ADDRESSED THIS QUESTION BY USING A TARGETED-CAPTURE APPROACH CALLED METHYL-SEQ TO INVESTIGATE DNA METHYLATION PATTERNS OF THE DENTATE GYRUS AT PUTATIVE REGULATORY REGIONS ACROSS THE MOUSE GENOME FROM MICE EXPOSED TO 14 DAYS OF SOCIAL DEFEAT. FINDINGS WERE REPLICATED IN INDEPENDENT COHORTS BY BISULFITE-PYROSEQUENCING. TWO DIFFERENTIALLY METHYLATED REGIONS (DMRS) WERE IDENTIFIED. ONE DMR WAS LOCATED AT INTRON 9 OF DROSHA, AND IT SHOWED REDUCED METHYLATION IN STRESSED MICE. THIS OBSERVATION REPLICATED IN ONE OF TWO INDEPENDENT COHORTS. A SECOND DMR WAS IDENTIFIED AT AN INTERGENIC REGION OF CHROMOSOME X, AND METHYLATION IN THIS REGION WAS INCREASED IN STRESSED MICE. THIS METHYLATION DIFFERENCE REPLICATED IN TWO INDEPENDENT COHORTS AND IN MAJOR DEPRESSIVE DISORDER (MDD) POSTMORTEM BRAINS. THESE RESULTS HIGHLIGHT A REGION NOT PREVIOUSLY KNOWN TO BE DIFFERENTIALLY METHYLATED BY CHRONIC SOCIAL DEFEAT STRESS AND WHICH MAY BE INVOLVED IN MDD. 2018