1 2046 110 EPIGENETIC CLOCK INDICATES ACCELERATED AGING IN GLIAL CELLS OF PROGRESSIVE MULTIPLE SCLEROSIS PATIENTS. BACKGROUND: MULTIPLE SCLEROSIS (MS) IS A CHRONIC INFLAMMATORY NEURODEGENERATIVE DISEASE OF THE CENTRAL NERVOUS SYSTEM (CNS) CHARACTERIZED BY IRREVERSIBLE DISABILITY AT LATER PROGRESSIVE STAGES. A GROWING BODY OF EVIDENCE SUGGESTS THAT DISEASE PROGRESSION DEPENDS ON AGE AND INFLAMMATION WITHIN THE CNS. WE AIMED TO INVESTIGATE EPIGENETIC AGING IN BULK BRAIN TISSUE AND SORTED NUCLEI FROM MS PATIENTS USING DNA METHYLATION-BASED EPIGENETIC CLOCKS. METHODS: WE APPLIED HORVATH'S MULTI-TISSUE AND SHIREBY'S BRAIN-SPECIFIC CORTICAL CLOCK ON BULK BRAIN TISSUE (N = 46), SORTED NEURONAL (N = 54), AND GLIAL NUCLEI (N = 66) FROM POST-MORTEM BRAIN TISSUE OF PROGRESSIVE MS PATIENTS AND CONTROLS. RESULTS: WE FOUND A SIGNIFICANT INCREASE IN AGE ACCELERATION RESIDUALS, CORRESPONDING TO 3.6 YEARS, IN GLIAL CELLS OF MS PATIENTS COMPARED TO CONTROLS (P = 0.0024) USING THE CORTICAL CLOCK, WHICH HELD AFTER ADJUSTMENT FOR COVARIATES (P (ADJ) = 0.0263). THE 4.8-YEAR AGE ACCELERATION FOUND IN MS NEURONS (P = 0.0054) DID NOT WITHSTAND ADJUSTMENT FOR COVARIATES AND NO SIGNIFICANT DIFFERENCE IN AGE ACCELERATION RESIDUALS WAS OBSERVED IN BULK BRAIN TISSUE BETWEEN MS PATIENTS AND CONTROLS. CONCLUSION: WHILE THE FINDINGS WARRANT REPLICATION IN LARGER COHORTS, OUR STUDY SUGGESTS THAT GLIAL CELLS OF PROGRESSIVE MS PATIENTS EXHIBIT ACCELERATED BIOLOGICAL AGING. 2022 2 1537 29 DNA METHYLATION IN ADOLESCENTS WITH ANXIETY DISORDER: A LONGITUDINAL STUDY. ANXIETY DISORDERS (AD) TYPICALLY MANIFEST IN CHILDREN AND ADOLESCENTS AND MIGHT PERSIST INTO ADULTHOOD. HOWEVER, THERE ARE STILL FEW DATA CONCERNING EPIGENETIC MECHANISMS ASSOCIATED WITH ONSET, PERSISTENCE OR REMISSION OF AD OVER TIME. WE INVESTIGATED A COHORT OF ADOLESCENTS AND YOUNG ADULTS AT BASELINE (AGE; 13.19 +/- 2.38) AND AFTER 5 YEARS AND CLASSIFIED THEM ACCORDING TO THE AD DIAGNOSIS AND THEIR LONGITUDINAL TRAJECTORIES INTO 4 GROUPS: (1) TYPICALLY DEVELOPING COMPARISONS (TDC; CONTROL GROUP, N = 14); (2) INCIDENT (AD IN THE SECOND EVALUATION ONLY, N = 11); (3) PERSISTENT (AD IN BOTH EVALUATIONS, N = 14) AND (4) REMITTENT (AD IN THE FIRST EVALUATION ONLY, N = 8). DNA METHYLATION WAS EVALUATED WITH THE INFINIUM HUMANMETHYLATION450 BEADCHIP FROM SALIVA SAMPLES COLLECTED AT BOTH EVALUATIONS. GENE SET ENRICHMENT ANALYSIS WAS APPLIED TO CONSIDER BIOLOGICAL PATHWAYS. WE FOUND DECREASED DNA METHYLATION IN TDC GROUP WHILE THE CHRONIC CASES OF AD PRESENTED HYPERMETHYLATION IN CENTRAL NERVOUS SYSTEM DEVELOPMENT PATHWAYS. MOREOVER, WE SHOWED THAT THIS PERSISTENT GROUP ALSO PRESENTED HYPERMETHYLATION WHILE THE OTHER THREE GROUPS WERE ASSOCIATED WITH HYPOMETHYLATION IN NERVOUS SYSTEM DEVELOPMENT PATHWAY. INCIDENCE AND REMISSION GROUPS WERE ASSOCIATED WITH INCREASED AND DECREASED METHYLATION IN NEURON DEVELOPMENT PATHWAYS, RESPECTIVELY. LARGER STUDIES ARE LIKELY TO DETECT SPECIFIC GENES RELEVANT TO AD. 2018 3 177 31 ACCELERATED EPIGENETIC AGING AND INFLAMMATORY/IMMUNOLOGICAL PROFILE (IPAGE) IN PATIENTS WITH CHRONIC KIDNEY DISEASE. CHRONIC KIDNEY DISEASE (CKD) IS DEFINED BY A REDUCED ESTIMATED GLOMERULAR FILTRATION RATE (EGFR). THIS FAILURE CAN BE RELATED TO A PHENOTYPE OF ACCELERATED AGING. IN THIS WORK, WE CONSIDERED 76 PATIENTS WITH END-STAGE RENAL DISEASE (ESRD) AND 83 HEALTHY CONTROLS. WE CONCOMITANTLY EVALUATED FOR THE FIRST TIME TWO MEASURES THAT CAN BE INFORMATIVE OF THE RATE OF AGING, I.E., WHOLE BLOOD DNA METHYLATION USING THE ILLUMINA INFINIUM EPIC ARRAY AND PLASMA LEVELS OF A SELECTION OF INFLAMMATORY/IMMUNOLOGICAL PROTEINS USING MULTIPLEX IMMUNOASSAYS. FIRST OF ALL, WE DEMONSTRATED ACCELERATED AGING IN TERMS OF THE MOST COMMON EPIGENETIC AGE ESTIMATORS IN CKD PATIENTS. MOREOVER, WE DEVELOPED A NEW CLOCK/PREDICTOR OF AGE BASED ON THE INFLAMMATORY/IMMUNOLOGICAL PROFILE (IPAGE) AND IDENTIFIED THE INFLAMMATORY/IMMUNOLOGICAL BIOMARKERS DIFFERENTIALLY EXPRESSED BETWEEN CASES AND CONTROLS. IPAGE APPEARED TO BE MORE SENSITIVE THAN EPIGENETIC CLOCKS IN QUANTIFYING THE ACCELERATED AGING PHENOTYPE OF ESRD PATIENTS. INTERESTINGLY, WE DID NOT FIND ANY CORRELATION BETWEEN THE AGE ACCELERATION EVALUATED ACCORDING TO THE EPIGENETIC CLOCKS AND IPAGE IN EITHER THE ESRD GROUP OR THE CONTROL GROUP. ON THE WHOLE, OUR DATA SHOW A CONSISTENT ACCELERATED AGING PHENOTYPE IN ESRD PATIENTS, WHICH IS BETTER APPRECIATED BY QUANTIFYING THE UNDERLYING INFLAMMATORY PROCESSES (INFLAMMAGING) BY IPAGE THAN BY USING EPIGENETIC CLOCKS. 2022 4 1526 44 DNA METHYLATION CHANGES IN GLIAL CELLS OF THE NORMAL-APPEARING WHITE MATTER IN MULTIPLE SCLEROSIS PATIENTS. MULTIPLE SCLEROSIS (MS), THE LEADING CAUSE OF NON-TRAUMATIC NEUROLOGICAL DISABILITY IN YOUNG ADULTS, IS A CHRONIC INFLAMMATORY AND NEURODEGENERATIVE DISEASE OF THE CENTRAL NERVOUS SYSTEM (CNS). DUE TO THE POOR ACCESSIBILITY TO THE TARGET ORGAN, CNS-CONFINED PROCESSES UNDERPINNING THE LATER PROGRESSIVE FORM OF MS REMAIN ELUSIVE THEREBY LIMITING TREATMENT OPTIONS. WE AIMED TO EXAMINE DNA METHYLATION, A STABLE EPIGENETIC MARK OF GENOME ACTIVITY, IN GLIAL CELLS TO CAPTURE RELEVANT MOLECULAR CHANGES UNDERLYING MS NEUROPATHOLOGY. WE PROFILED DNA METHYLATION IN NUCLEI OF NON-NEURONAL CELLS, ISOLATED FROM 38 POST-MORTEM NORMAL-APPEARING WHITE MATTER (NAWM) SPECIMENS OF MS PATIENTS (N = 8) IN COMPARISON TO WHITE MATTER OF CONTROL INDIVIDUALS (N = 14), USING INFINIUM METHYLATIONEPIC BEADCHIP. WE IDENTIFIED 1,226 SIGNIFICANT (GENOME-WIDE ADJUSTED P-VALUE < 0.05) DIFFERENTIALLY METHYLATED POSITIONS (DMPS) BETWEEN MS PATIENTS AND CONTROLS. FUNCTIONAL ANNOTATION OF THE ALTERED DMP-GENES UNCOVERED ALTERATIONS OF PROCESSES RELATED TO CELLULAR MOTILITY, CYTOSKELETON DYNAMICS, METABOLIC PROCESSES, SYNAPTIC SUPPORT, NEUROINFLAMMATION AND SIGNALING, SUCH AS WNT AND TGF-BETA PATHWAYS. A FRACTION OF THE AFFECTED GENES DISPLAYED TRANSCRIPTIONAL DIFFERENCES IN THE BRAIN OF MS PATIENTS, AS REPORTED BY PUBLICALLY AVAILABLE TRANSCRIPTOMIC DATA. CELL TYPE-RESTRICTED ANNOTATION OF DMP-GENES ATTRIBUTED ALTERATIONS OF CYTOSKELETON REARRANGEMENT AND EXTRACELLULAR MATRIX REMODELLING TO ALL GLIAL CELL TYPES, WHILE SOME PROCESSES, INCLUDING ION TRANSPORT, WNT/TGF-BETA SIGNALING AND IMMUNE PROCESSES WERE MORE SPECIFICALLY LINKED TO OLIGODENDROCYTES, ASTROCYTES AND MICROGLIAL CELLS, RESPECTIVELY. OUR FINDINGS STRONGLY SUGGEST THAT NAWM GLIAL CELLS ARE HIGHLY ALTERED, EVEN IN THE ABSENCE OF LESIONAL INSULT, COLLECTIVELY EXHIBITING A MULTICELLULAR REACTION IN RESPONSE TO DIFFUSE INFLAMMATION. 2022 5 1607 31 DNA METHYLATION, COLON CANCER AND MEDITERRANEAN DIET: RESULTS FROM THE EPIC-ITALY COHORT. THE BIOLOGICAL MECHANISMS THROUGH WHICH ADHERENCE TO MEDITERRANEAN DIET (MD) PROTECTS AGAINST COLON CANCER (CC) ARE POORLY UNDERSTOOD. EVIDENCE SUGGESTS THAT CHRONIC INFLAMMATION MAY BE IMPLICATED IN THE PATHWAY. BOTH DIET AND CC ARE RELATED TO EPIGENETIC REGULATION. WE PERFORMED A NESTED CASE-CONTROL STUDY ON 161 PAIRS FROM THE ITALIAN COMPONENT OF THE EUROPEAN PROSPECTIVE INVESTIGATION INTO CANCER AND NUTRITION (EPIC) COHORT, IN WHICH WE LOOKED FOR THE METHYLATION SIGNALS IN DNA EXTRACTED FROM LEUCOCYTES ASSOCIATED WITH BOTH CC AND MD IN 995 CPGS LOCATED IN 48 INFLAMMATION GENES. THE DNA METHYLATION SIGNALS DETECTED IN THIS ANALYSIS WERE VALIDATED IN A SUBGROUP OF 47 CASE-CONTROL PAIRS AND FURTHER REPLICATED (WHERE VALIDATED) IN 95 NEW PAIRS BY MEANS OF PYROSEQUENCING. AMONG THE CPG SITES SELECTED A-PRIORI IN INFLAMMATION-RELATED GENES, SEVEN CPG SITES WERE FOUND TO BE ASSOCIATED WITH CC STATUS AND WITH MD, IN LINE WITH ITS PROTECTIVE EFFECT. ONLY TWO CPG SITES (CG17968347-SERPINE1 AND CG20674490-RUNX3) WERE VALIDATED USING BISULPHITE PYROSEQUENCING AND, AFTER REPLICATION, WE FOUND THAT DNA METHYLATION OF CG20674490-RUNX3 MAY BE A POTENTIAL MOLECULAR MEDIATOR EXPLAINING THE PROTECTIVE EFFECT OF MD ON CC ONSET. THE USE OF A 'MEET-IN-THE-MIDDLE' APPROACH TO IDENTIFY THE OVERLAP BETWEEN EXPOSURE AND PREDICTIVE MARKERS OF DISEASE IS INNOVATIVE IN STUDIES ON THE RELATIONSHIP BETWEEN DIET AND CANCER, IN WHICH EXPOSURE ASSESSMENT IS DIFFICULT AND THE MECHANISMS THROUGH WHICH THE NUTRIENTS EXERT THEIR PROTECTIVE EFFECT IS LARGELY UNKNOWN. 2019 6 4024 39 LUNG ALLOGRAFT EPITHELIUM DNA METHYLATION AGE IS ASSOCIATED WITH GRAFT CHRONOLOGIC AGE AND PRIMARY GRAFT DYSFUNCTION. ADVANCED DONOR AGE IS A RISK FACTOR FOR POOR SURVIVAL FOLLOWING LUNG TRANSPLANTATION. HOWEVER, RECENT WORK IDENTIFYING EPIGENETIC DETERMINANTS OF AGING HAS SHOWN THAT BIOLOGIC AGE MAY NOT ALWAYS REFLECT CHRONOLOGIC AGE AND THAT STRESSORS CAN ACCELERATE BIOLOGIC AGING. WE HYPOTHESIZED THAT LUNG ALLOGRAFTS THAT EXPERIENCED PRIMARY GRAFT DYSFUNCTION (PGD), CHARACTERIZED BY POOR OXYGENATION IN THE FIRST THREE POST-TRANSPLANT DAYS, WOULD HAVE INCREASED BIOLOGIC AGE. WE CULTURED AIRWAY EPITHELIAL CELLS ISOLATED BY TRANSBRONCHIAL BRUSH AT 1-YEAR BRONCHOSCOPIES FROM 13 SUBJECTS WITH SEVERE PGD AND 15 CONTROLS MATCHED ON AGE AND TRANSPLANT INDICATION. WE MEASURED EPIGENETIC AGE USING THE HORVATH EPIGENETIC CLOCK. LINEAR MODELS WERE USED TO DETERMINE THE ASSOCIATION OF AIRWAY EPIGENETIC AGE WITH CHRONOLOGIC AGES AND PGD STATUS, ADJUSTED FOR RECIPIENT PGD RISK FACTORS. SURVIVAL MODELS ASSESSED THE ASSOCIATION WITH CHRONIC LUNG ALLOGRAFT DYSFUNCTION (CLAD) OR DEATH. DISTRIBUTIONS OF PROMOTER METHYLATION WITHIN PATHWAYS WERE COMPARED BETWEEN GROUPS. DNA METHYLTRANSFERASE (DNMT) ACTIVITY WAS QUANTIFIED IN AIRWAY EPITHELIAL CELLS UNDER HYPOXIC OR NORMOXIC CONDITIONS. AIRWAY EPIGENETIC AGE APPEARED YOUNGER BUT WAS STRONGLY ASSOCIATED WITH THE AGE OF THE ALLOGRAFT (SLOPE 0.38 PER YEAR, 95% CI 0.27-0.48). THERE WAS NO CORRELATION BETWEEN EPIGENETIC AGE AND RECIPIENT AGE (P = 0.96). EPIGENETIC AGE WAS 6.5 YEARS GREATER (95% CI 1.7-11.2) IN SUBJECTS WHO HAD EXPERIENCED PGD, AND THIS EFFECT REMAINED SIGNIFICANT AFTER ADJUSTING FOR DONOR AND RECIPIENT CHARACTERISTICS (P = 0.03). EPIGENETIC AGE WAS NOT ASSOCIATED WITH CLAD-FREE SURVIVAL RISK (P = 0.11). ANALYSIS OF DIFFERENTIAL METHYLATION OF PROMOTERS OF KEY BIOLOGIC PATHWAYS REVEALED HYPOMETHYLATION IN REGIONS RELATED TO HYPOXIA, INFLAMMATION, AND METABOLISM-ASSOCIATED PATHWAYS. ACCORDINGLY, AIRWAY EPITHELIAL CELLS CULTURED IN HYPOXIC CONDITIONS SHOWED SUPPRESSED DNMT ACTIVITY. WHILE AIRWAY METHYLATION AGE WAS PRIMARILY DETERMINED BY DONOR CHRONOLOGIC AGE, EARLY INJURY IN THE FORM OF PGD WAS ASSOCIATED WITH INCREASED ALLOGRAFT EPIGENETIC AGE. THESE DATA SHOW HOW PGD MIGHT SUPPRESS KEY PROMOTER METHYLATION RESULTING IN LONG-TERM IMPACTS ON THE ALLOGRAFT. 2021 7 175 27 ACCELERATED AGING WITH HIV BEGINS AT THE TIME OF INITIAL HIV INFECTION. LIVING WITH HIV INFECTION IS ASSOCIATED WITH EARLY ONSET OF AGING-RELATED CHRONIC CONDITIONS, SOMETIMES DESCRIBED AS ACCELERATED AGING. EPIGENETIC DNA METHYLATION PATTERNS CAN EVALUATE ACCELERATION OF BIOLOGICAL AGE RELATIVE TO CHRONOLOGICAL AGE. THE IMPACT OF INITIAL HIV INFECTION ON FIVE EPIGENETIC MEASURES OF AGING WAS EXAMINED BEFORE AND APPROXIMATELY 3 YEARS AFTER HIV INFECTION IN THE SAME INDIVIDUALS (N=102). SIGNIFICANT EPIGENETIC AGE ACCELERATION (MEDIAN 1.9-4.8 YEARS) AND ESTIMATED TELOMERE LENGTH SHORTENING (ALL P