1 4008 114 LOW DOSE OF URANIUM INDUCES MULTIGENERATIONAL EPIGENETIC EFFECTS IN RAT KIDNEY. PURPOSE: A PROTOCOL OF CHRONIC EXPOSURE TO LOW DOSE OF URANIUM WAS ESTABLISHED IN ORDER TO DISTINGUISH THE SEXUAL DIFFERENCES AND THE DEVELOPMENTAL PROCESS THAT ARE CRITICAL WINDOWS FOR EPIGENETIC EFFECTS OVER GENERATIONS. METHODS: BOTH MALE AND FEMALE RATS WERE CONTAMINATED THROUGH THEIR DRINKING WATER WITH A NON-TOXIC SOLUTION OF URANYL NITRATE FOR 9 MONTHS. THE EXPOSED GENERATION (F0) AND THE FOLLOWING TWO GENERATIONS (F1 AND F2) WERE EXAMINED. CLINICAL MONITORING, GLOBAL DNA METHYLATION PROFILE AND DNA METHYLTRANSFERASES (DNMTS) GENE EXPRESSION WERE ANALYZED IN KIDNEYS. RESULTS: WHILE THE BODY WEIGHT OF F1 MALES INCREASED, A SMALL DECREASE IN KIDNEY AND BODY WEIGHT WAS OBSERVED IN F2 MALES. IN ADDITION, GLOBAL DNA HYPERMETHYLATION PROFILE IN KIDNEY CELLS WAS OBSERVED IN F1 AND F2 MALES. QPCR RESULTS REVEAL A SIGNIFICANT INCREASE OF METHYLTRANSFERASE GENES EXPRESSION (DNMT1 AND DNMT3A) FOR F2 FEMALES. CONCLUSIONS: IN THE FIELD OF PUBLIC HEALTH POLICY AND TO RAISE ATTENTION TO GENERATIONAL EFFECTS FOR THE RISK ASSESSMENT OF THE ENVIRONMENTAL EXPOSURES, LOW DOSES OF URANIUM DO NOT IMPLY CLINICAL EFFECTS ON ADULT EXPOSED RATS. HOWEVER, OUR RESULTS CONFIRM THE IMPORTANCE OF THE DEVELOPMENTAL WINDOWS' SENSITIVITY IN ADDITION TO THE SEXUAL DIMORPHISMS OF THE OFFSPRING. 2018 2 457 55 APPLYING A MULTISCALE SYSTEMS BIOLOGY APPROACH TO STUDY THE EFFECT OF CHRONIC LOW-DOSE EXPOSURE TO URANIUM IN RAT KIDNEYS. PURPOSE: TO EXAMINE THE EFFECTS OF LOW-DOSE EXPOSURE TO URANIUM WITH A SYSTEMS BIOLOGY APPROACH, A MULTISCALE HIGH-THROUGHPUT MULTI-OMICS ANALYSIS WAS APPLIED WITH A PROTOCOL FOR CHRONIC EXPOSURE TO THE RAT KIDNEY. METHODS: MALE AND FEMALE RATS WERE CONTAMINATED FOR NINE MONTHS THROUGH THEIR DRINKING WATER WITH A NONTOXIC SOLUTION OF URANYL NITRATE. A MULTISCALE APPROACH ENABLED CLINICAL MONITORING ASSOCIATED WITH METABOLOMIC AND TRANSCRIPTOMIC (MRNA AND MICRORNA) ANALYSES. RESULTS: A SEX-INTERACTION EFFECT WAS OBSERVED IN THE KIDNEY, URINE, AND PLASMA METABOLOMES OF CONTAMINATED RATS. MOREOVER, URINE AND KIDNEY METABOLIC PROFILES CORRELATED AND CONFIRMED THAT THE PRIMARY DYSREGULATED METABOLISMS ARE THOSE OF NICOTINATE-NICOTINAMIDE AND OF UNSATURATED FATTY ACID BIOSYNTHESIS. UPSTREAM OF THE METABOLIC PATHWAYS, TRANSCRIPTOMIC PROFILES OF THE KIDNEY REVEAL GENE ACTIVITY FOCUSED ON GENE REGULATION MECHANISMS, CELL SIGNALING, CELL STRUCTURE, DEVELOPMENTAL PROCESSES, AND CELL PROLIFERATION. EXAMINATION OF EPIGENETIC POST-TRANSCRIPTIONAL GENE REGULATION PROCESSES SHOWED SIGNIFICANT DYSREGULATION OF 70 MICRO-RNAS. THE MULTI-OMICS APPROACH HIGHLIGHTED THE ACTIVITIES OF THE CELLS' BIOLOGICAL PROCESSES ON MULTIPLE SCALES THROUGH ANALYSIS OF GENE EXPRESSION, CONFIRMED BY CHANGES OBSERVED IN THE METABOLOME. CONCLUSION: OUR RESULTS SHOWED CHANGES IN MULTI-OMIC PROFILES OF RATS EXPOSED TO LOW DOSES OF URANIUM CONTAMINATION, COMPARED WITH CONTROLS. THESE CHANGES INVOLVED GENE EXPRESSION AS WELL AS MODIFICATIONS IN THE TRANSCRIPTOME AND THE METABOLOME. THE METABOLOMIC PROFILE CONFIRMED THAT THE MAIN MOLECULAR TARGETS OF URANIUM IN KIDNEY CELLS ARE THE METABOLISM OF NICOTINATE-NICOTINAMIDE AND THE BIOSYNTHESIS OF UNSATURATED FATTY ACIDS. ADDITIONALLY, GENE EXPRESSION ANALYSIS SHOWED THAT THE METABOLISM OF FATTY ACIDS IS TARGETED BY PROCESSES ASSOCIATED WITH CELL FUNCTION. THESE RESULTS DEMONSTRATE THAT MULTISCALE SYSTEMS BIOLOGY IS USEFUL IN ELUCIDATING THE MOST DISCRIMINATIVE PATHWAYS FROM GENOMIC TO METABOLOMIC LEVELS FOR ASSESSING THE BIOLOGICAL IMPACT OF THIS LOW-LEVEL ENVIRONMENTAL EXPOSURE, I.E. THE EXPOSOME. 2019 3 1511 58 DNA METHYLATION AND POTENTIAL MULTIGENERATIONAL EPIGENETIC EFFECTS LINKED TO URANIUM CHRONIC LOW-DOSE EXPOSURE IN GONADS OF MALES AND FEMALES RATS. INTRODUCTION: AN INCREASED HEALTH PROBLEM IN INDUSTRIALISED COUNTRIES IS THE CONTEMPORARY CONCERN OF PUBLIC AND SCIENTIFIC COMMUNITY AS WELL. THIS HAS BEEN ATTRIBUTED IN PART TO ACCUMULATED ENVIRONMENTAL POLLUTANTS ESPECIALLY RADIOACTIVE SUBSTANCES AND THE USE OF NUCLEAR POWER PLANTS WORLDWIDE. HOWEVER, THE OUTCOME OF CHRONIC EXPOSURE TO LOW DOSES OF A RADIONUCLIDE SUCH AS URANIUM REMAINS UNKNOWN. RECENTLY, A PARADIGM SHIFT IN THE PERCEPTION OF RISK OF RADIOTOXICOLOGY HAS EMERGED THROUGH INVESTIGATING THE POSSIBILITY OF TRANSMISSION OF BIOLOGICAL EFFECTS OVER GENERATIONS, IN PARTICULAR BY EPIGENETIC PATHWAYS. THESE PROCESSES ARE KNOWN FOR THEIR CRUCIAL ROLES ASSOCIATED WITH THE DEVELOPMENT OF SEVERAL DISEASES. OBJECTIVE: THE CURRENT WORK INVESTIGATES THE EPIGENETIC EFFECT OF CHRONIC EXPOSURE TO LOW DOSES OF URANIUM AND ITS INHERITANCE ACROSS GENERATIONS. MATERIALS AND METHODS TO TEST THIS PROPOSITION, A RODENT MULTIGENERATIONAL MODEL, MALES AND FEMALES, WERE EXPOSED TO A NON-TOXIC CONCENTRATION OF URANIUM (40MGL(-1) DRINKING WATER) FOR NINE MONTHS. THE URANIUM EFFECTS ON WERE EVALUATED OVER THREE GENERATIONS (F0, F1 AND F2) BY ANALYSING THE DNA METHYLATION PROFILE AND DNMT GENES EXPRESSION IN OVARIES AND TESTES TISSUES. RESULTS: HERE WE REPORT A SIGNIFICANT HYPERMETHYLATION OF TESTES DNA (P <0.005) WHEREAS OVARIES SHOWED HYPOMETHYLATED DNA (P <0.005). INTERESTINGLY, THIS DNA METHYLATION PROFILE WAS SIGNIFICANTLY MAINTAINED ACROSS GENERATIONS F0, F1 AND F2. FURTHERMORE, QPCR RESULTS OF BOTH TISSUES IMPLY A SIGNIFICANT CHANGE IN THE EXPRESSION OF DNA METHYLTRANSFERASE GENES (DNMT 1 AND DNMT3A/B) AS WELL. CONCLUSION: ALTOGETHER, OUR WORK DEMONSTRATES FOR THE FIRST TIME A SEX-DEPENDANCE AND INHERITANCE OF EPIGENETIC MARKS, DNA METHYLATION, AS A BIOLOGICAL RESPONSE TO THE EXPOSURE TO LOW DOSES OF URANIUM. HOWEVER, IT IS NOT CLEAR WHICH TYPE OF REPRODUCTIVE CELL TYPE IS MORE RESPONSIVE IN THIS CONTEXT. 2018 4 3300 38 HIGH-FAT DIET REPROGRAMS THE EPIGENOME OF RAT SPERMATOZOA AND TRANSGENERATIONALLY AFFECTS METABOLISM OF THE OFFSPRING. OBJECTIVES: CHRONIC AND HIGH CONSUMPTION OF FAT CONSTITUTES AN ENVIRONMENTAL STRESS THAT LEADS TO METABOLIC DISEASES. WE HYPOTHESIZED THAT HIGH-FAT DIET (HFD) TRANSGENERATIONALLY REMODELS THE EPIGENOME OF SPERMATOZOA AND METABOLISM OF THE OFFSPRING. METHODS: F0-MALE RATS FED EITHER HFD OR CHOW DIET FOR 12 WEEKS WERE MATED WITH CHOW-FED DAMS TO GENERATE F1 AND F2 OFFSPRING. MOTILE SPERMATOZOA WERE ISOLATED FROM F0 AND F1 BREEDERS TO DETERMINE DNA METHYLATION AND SMALL NON-CODING RNA (SNCRNA) EXPRESSION PATTERN BY DEEP SEQUENCING. RESULTS: NEWBORN OFFSPRING OF HFD-FED FATHERS HAD REDUCED BODY WEIGHT AND PANCREATIC BETA-CELL MASS. ADULT FEMALE, BUT NOT MALE, OFFSPRING OF HFD-FED FATHERS WERE GLUCOSE INTOLERANT AND RESISTANT TO HFD-INDUCED WEIGHT GAIN. THIS PHENOTYPE WAS PERPETUATED IN THE F2 PROGENY, INDICATING TRANSGENERATIONAL EPIGENETIC INHERITANCE. THE EPIGENOME OF SPERMATOZOA FROM HFD-FED F0 AND THEIR F1 MALE OFFSPRING SHOWED COMMON DNA METHYLATION AND SMALL NON-CODING RNA EXPRESSION SIGNATURES. ALTERED EXPRESSION OF SPERM MIRNA LET-7C WAS PASSED DOWN TO METABOLIC TISSUES OF THE OFFSPRING, INDUCING A TRANSCRIPTOMIC SHIFT OF THE LET-7C PREDICTED TARGETS. CONCLUSION: OUR RESULTS PROVIDE INSIGHT INTO MECHANISMS BY WHICH HFD TRANSGENERATIONALLY REPROGRAMS THE EPIGENOME OF SPERM CELLS, THEREBY AFFECTING METABOLIC TISSUES OF OFFSPRING THROUGHOUT TWO GENERATIONS. 2016 5 73 38 A MULTI-GENERATIONAL STUDY ON LOW-DOSE BPA EXPOSURE IN WISTAR RATS: EFFECTS ON MATERNAL BEHAVIOR, FLAVOR INTAKE AND DEVELOPMENT. BISPHENOL A (BPA) IS A COMMON ENDOCRINE DISRUPTOR FOUND AS AN ENVIRONMENTAL AND FOOD CONTAMINANT. IT EXERTS BOTH DEVELOPMENTAL AND BEHAVIORAL EFFECTS, MAINLY WHEN EXPOSURE OCCURS IN EARLY LIFE. THE AIM OF THIS STUDY WAS TO DETERMINE THE MULTI-GENERATIONAL EFFECTS OF CHRONIC, HUMAN-RELEVANT LOW-DOSE EXPOSURE TO BPA ON DEVELOPMENT, MATERNAL BEHAVIOR AND FLAVOR PREFERENCE IN WISTAR RATS. BPA WAS ORALLY ADMINISTERED AT A DAILY DOSE OF 5 MUG/KG BODY WEIGHT TO F0 PREGNANT DAMS FROM THE FIRST DAY OF GESTATION (GD 1) UNTIL THE LAST DAY OF LACTATION (LD 21), AND THEN TO F1 OFFSPRING FROM WEANING (PND 21) TO ADULTHOOD (PND 100). F2 OFFSPRING WERE NOT EXPOSED. DEVELOPMENT AND CLINICAL SIGNS OF TOXICITY WERE ASSESSED DAILY. MATERNAL BEHAVIOR WAS EVALUATED BY OBSERVING NURSING AND PUP-CARING ACTIONS, AS WELL AS "NON-MATERNAL" BEHAVIORS IN F0 AND F1 DAMS FROM PARTURITION UNTIL LD 8. THE FLAVOR PREFERENCES OF F1 AND F2 OFFSPRING WERE EVALUATED BASED ON THE INTAKE OF SWEET, SALT AND FAT SOLUTIONS USING THE TWO-BOTTLE CHOICE TEST ON PND 21-34 AND PND 86-99. BPA EXPOSURE: 1) DECREASED MATERNAL BEHAVIOR IN F1 DAMS, 2) CAUSED DEVELOPMENTAL DEFECTS IN BOTH F1 AND F2 OFFSPRING, WITH A NOTICEABLE DECREASE IN ANOGENITAL DISTANCE IN MALE RATS, AND 3) DID NOT AFFECT FLAVORED SOLUTION INTAKE IN F1, BUT INDUCED CHANGES IN SWEET PREFERENCE IN F2 JUVENILES AND IN SALT AND FAT SOLUTION INTAKES IN F2 ADULTS, AND 4) INDUCED A BODY WEIGHT INCREASE IN THE F2 GENERATION ONLY, WHEREAS FOOD INTAKE AND WATER CONSUMPTION DID NOT CHANGE. TAKEN AS A WHOLE, OUR FINDINGS SHOWED THAT BOTH GESTATIONAL (F0) AND LIFELONG (F1) EXPOSURES TO A HUMAN-RELEVANT DOSE OF BPA COULD INDUCE MULTI-GENERATIONAL EFFECTS ON BOTH DEVELOPMENT AND BEHAVIOR. THESE RESULTS SUGGEST POSSIBLE SELECTIVE NEUROENDOCRINE DEFECTS AND/OR EPIGENETIC CHANGES CAUSED BY BPA EXPOSURE. 2014 6 2985 26 GENETIC DIVERSITY FOR NITROGEN USE EFFICIENCY IN ARABIDOPSIS THALIANA. THE PLASTICITY OF PLANT GROWTH RESPONSE TO DIFFERING NITRATE AVAILABILITY RENDERS THE IDENTIFICATION OF BIOMARKERS DIFFICULT, BUT ALLOWS ACCESS TO GENETIC FACTORS AS TOOLS TO MODULATE ROOT SYSTEMS TO A WIDE RANGE OF SOIL CONDITIONS. NITROGEN AVAILABILITY IS A MAJOR DETERMINANT OF CROP YIELD. WHILE THE APPLICATION OF FERTILISER SUBSTANTIALLY INCREASES THE YIELD ON POOR SOILS, IT ALSO CAUSES NITRATE POLLUTION OF WATER RESOURCES AND HIGH COSTS FOR FARMERS. INCREASING NITROGEN USE EFFICIENCY IN CROP PLANTS IS A NECESSARY STEP TO IMPLEMENT LOW-INPUT AGRICULTURAL SYSTEMS. WE EXPLOITED THE GENETIC DIVERSITY PRESENT IN THE WORLDWIDE ARABIDOPSIS THALIANA POPULATION TO STUDY ADAPTIVE GROWTH PATTERNS AND CHANGES IN GENE EXPRESSION ASSOCIATED WITH CHRONIC LOW NITRATE STRESS, TO IDENTIFY BIOMARKERS ASSOCIATED WITH GOOD PLANT PERFORMANCE UNDER LOW NITRATE AVAILABILITY. ARABIDOPSIS ACCESSIONS WERE GROWN ON AGAR PLATES WITH LIMITED AND SUFFICIENT SUPPLY OF NITRATE TO MEASURE ROOT SYSTEM ARCHITECTURE AS WELL AS SHOOT AND ROOT FRESH WEIGHT. DIFFERENTIAL GENE EXPRESSION WAS DETERMINED USING AFFYMETRIX ATH1 ARRAYS. WE SHOW THAT THE RESPONSE TO DIFFERING NITRATE AVAILABILITY IS HIGHLY VARIABLE IN ARABIDOPSIS ACCESSIONS. ANALYSES OF VEGETATIVE SHOOT GROWTH AND ROOT SYSTEM ARCHITECTURE IDENTIFIED ACCESSION-SPECIFIC REACTION MODES TO COPE WITH LIMITED NITRATE AVAILABILITY. TRANSCRIPTION AND EPIGENETIC FACTORS WERE IDENTIFIED AS IMPORTANT PLAYERS IN THE ADAPTION TO LIMITED NITROGEN IN A GLOBAL GENE EXPRESSION ANALYSIS. FIVE NITRATE-RESPONSIVE GENES EMERGED AS POSSIBLE BIOMARKERS FOR NUE IN ARABIDOPSIS. THE PLASTICITY OF PLANT GROWTH IN RESPONSE TO DIFFERING NITRATE AVAILABILITY IN THE SUBSTRATE RENDERS THE IDENTIFICATION OF MORPHOLOGICAL AND MOLECULAR FEATURES AS BIOMARKERS DIFFICULT, BUT AT THE SAME TIME ALLOWS ACCESS TO A MULTITUDE OF GENETIC FACTORS WHICH CAN BE USED AS TOOLS TO MODULATE AND ADJUST ROOT SYSTEMS TO A WIDE RANGE OF SOIL CONDITIONS. 2019 7 1325 31 DEPLETED URANIUM INDUCES SEX- AND TISSUE-SPECIFIC METHYLATION PATTERNS IN ADULT ZEBRAFISH. WE EXAMINED THE EFFECTS OF CHRONIC EXPOSURE TO DIFFERENT CONCENTRATIONS (2 AND 20 MUG L(-)(1)) OF ENVIRONMENTALLY RELEVANT WATERBORNE DEPLETED URANIUM (DU) ON THE DNA METHYLATION PATTERNS BOTH AT HPAII RESTRICTION SITES (5'-CCGG-3') AND ACROSS THE WHOLE GENOME IN THE ZEBRAFISH BRAIN, GONADS, AND EYES. WE FIRST IDENTIFIED SEX-DEPENDENT DIFFERENCES IN THE METHYLATION LEVEL OF HPAII SITES AFTER EXPOSURE. IN MALES, THESE EFFECTS WERE PRESENT AS EARLY AS 7 DAYS AFTER EXPOSURE TO 20 MUG L(-)(1) DU, AND WERE EVEN MORE PRONOUNCED IN THE BRAIN, GONADS, AND EYES AFTER 24 DAYS. HOWEVER, IN FEMALES, HYPOMETHYLATION WAS ONLY OBSERVED IN THE GONADS AFTER EXPOSURE TO 20 MUG L(-)(1) DU FOR 24 DAYS. SEX-SPECIFIC EFFECTS OF DU WERE ALSO APPARENT AT THE WHOLE-GENOME LEVEL, BECAUSE IN MALES, EXPOSURE TO 20 MUG L(-)(1) DU FOR 24 DAYS RESULTED IN CYTOSINE HYPERMETHYLATION IN THE BRAIN AND EYES AND HYPOMETHYLATION IN THE GONADS. IN CONTRAST, IN FEMALES, HYPERMETHYLATION WAS OBSERVED IN THE BRAIN AFTER EXPOSURE TO BOTH CONCENTRATIONS OF DU FOR 7 DAYS. BASED ON OUR CURRENT KNOWLEDGE OF URANIUM TOXICITY, SEVERAL HYPOTHESES ARE PROPOSED TO EXPLAIN THESE FINDINGS, INCLUDING THE INVOLVEMENT OF OXIDATIVE STRESS, ALTERATION OF DEMETHYLATION ENZYMES AND THE CALCIUM SIGNALING PATHWAY. THIS STUDY REPORTS, FOR THE FIRST TIME, THE SEX- AND TISSUE-SPECIFIC EPIGENETIC CHANGES THAT OCCUR IN A NONHUMAN ORGANISM AFTER EXPOSURE TO ENVIRONMENTALLY RELEVANT CONCENTRATIONS OF URANIUM, WHICH COULD INDUCE TRANSGENERATIONAL EPIGENETIC EFFECTS. 2016 8 4231 28 METHYLATION OF PROTOCADHERIN 10, A NOVEL TUMOR SUPPRESSOR, IS ASSOCIATED WITH POOR PROGNOSIS IN PATIENTS WITH GASTRIC CANCER. BACKGROUND & AIMS: BY USING METHYLATION-SENSITIVE REPRESENTATIONAL DIFFERENCE ANALYSIS, WE IDENTIFIED PROTOCADHERIN 10 (PCDH10), A GENE THAT ENCODES A PROTOCADHERIN AND IS SILENCED IN A TUMOR-SPECIFIC MANNER. WE ANALYZED ITS EPIGENETIC INACTIVATION, BIOLOGICAL EFFECTS, AND PROGNOSTIC SIGNIFICANCE IN GASTRIC CANCER. METHODS: METHYLATION STATUS WAS EVALUATED BY COMBINED BISULFITE RESTRICTION ANALYSIS AND BISULFITE SEQUENCING. THE EFFECTS OF PCDH10 RE-EXPRESSION WERE DETERMINED IN GROWTH, APOPTOSIS, PROLIFERATION, AND INVASION ASSAYS. PCDH10 TARGET GENES WERE IDENTIFIED BY COMPLEMENTARY DNA MICROARRAY ANALYSIS. RESULTS: PCDH10 WAS SILENCED OR DOWN-REGULATED IN 94% (16 OF 17) OF GASTRIC CANCER CELL LINES; EXPRESSION LEVELS WERE RESTORED BY EXPOSURE TO DEMETHYLATING AGENTS. RE-EXPRESSION OF PCDH10 IN MKN45 GASTRIC CANCER CELLS REDUCED COLONY FORMATION IN VITRO AND TUMOR GROWTH IN MICE; IT ALSO INHIBITED CELL PROLIFERATION (P < .01), INDUCED CELL APOPTOSIS (P < .001), AND REPRESSED CELL INVASION (P < .05), UP-REGULATING THE PRO-APOPTOSIS GENES FAS, CASPASE 8, JUN, AND CDKN1A; THE ANTIPROLIFERATION GENE FGFR; AND THE ANTI-INVASION GENE HTATIP2. PCDH10 METHYLATION WAS DETECTED IN 82% (85 OF 104) OF GASTRIC TUMORS COMPARED WITH 37% (38 OF 104) OF PAIRED NONTUMOR TISSUES (P < .0001). IN THE LATTER, PCDH10 METHYLATION WAS HIGHER IN PRECANCEROUS LESIONS (27 OF 45; 60%) THAN IN CHRONIC GASTRITIS SAMPLES (11 OF 59; 19%) (P < .0001). AFTER A MEDIAN FOLLOW-UP PERIOD OF 16.8 MONTHS, MULTIVARIATE ANALYSIS REVEALED THAT PATIENTS WITH PCDH10 METHYLATION IN ADJACENT NONTUMOR AREAS HAD A SIGNIFICANT DECREASE IN OVERALL SURVIVAL. KAPLAN-MEIER SURVIVAL CURVES SHOWED THAT PCDH10 METHYLATION WAS ASSOCIATED SIGNIFICANTLY WITH SHORTENED SURVIVAL IN STAGE I-III GASTRIC CANCER PATIENTS. CONCLUSIONS: PCDH10 IS A GASTRIC TUMOR SUPPRESSOR; ITS METHYLATION AT EARLY STAGES OF GASTRIC CARCINOGENESIS IS AN INDEPENDENT PROGNOSTIC FACTOR. 2009 9 4245 29 METHYLATION STATUS OF DDIT3 GENE IN CHRONIC MYELOID LEUKEMIA. BACKGROUND: DNA-DAMAGE-INDUCIBLE TRANSCRIPT 3 (DDIT3), A CANDIDATE TUMOR SUPPRESSOR GENE (TSG), HAS BEEN FOUND INVOLVED IN THE REGULATION OF CELLULAR GROWTH AND DIFFERENTIATION. THE EPIGENETIC CHANGES OF TSGS ARE RECENTLY RECOGNIZED AS AN ABNORMAL MECHANISM CONTRIBUTING TO THE DEVELOPMENT OF CHRONIC MYELOID LEUKEMIA (CML). THE AIM OF THIS STUDY WAS TO INVESTIGATE THE METHYLATION STATUS OF DDIT3 GENE IN CML PATIENTS. METHODS: THE METHYLATION STATUS OF DDIT3 PROMOTER WAS DETECTED IN THE BONE MARROW MONONUCLEAR CELLS FROM 53 PATIENTS WITH CML USING METHYLATION-SPECIFIC PCR (MSP). THE EXPRESSION LEVELS OF DDIT3 AND BCR/ABL TRANSCRIPT WERE DETERMINED BY REAL-TIME QUANTITATIVE PCR (RQ-PCR). CLINICAL DATA OF THESE PATIENTS WERE COLLECTED AND ANALYZED. RESULTS: THE ABERRANT METHYLATION OF DDIT3 GENE PROMOTER WAS FOUND IN 35 OF 53 (66%) CML CASES. CORRELATION WAS NOT FOUND BETWEEN DDIT3 PROMOTER HYPERMETHYLATION AND THE AGE, SEX, HEMOGLOBIN CONCENTRATION, PLATELET COUNTS, CHROMOSOMAL ABNORMALITIES, BCR/ABL TRANSCRIPT, AND STAGING OF CML PATIENTS (P > 0.05), BUT FOUND BETWEEN DDIT3 PROMOTER HYPERMETHYLATION AND WBC COUNTS OF CML CASES (R = 0.781, P < 0.001). THE LEVEL OF DDIT3 TRANSCRIPT IN CML PATIENTS WAS SIGNIFICANTLY LOWER THAN THAT IN CONTROLS (MEDIAN 3.28 VS 19.69, P < 0.001), HOWEVER, THERE WAS NO DIFFERENCE IN THE LEVEL OF DDIT3 TRANSCRIPT BETWEEN METHYLATION-POSITIVE CML CASES (0.05-65.32, MEDIAN 2.13) AND METHYLATION- NEGATIVE CML CASES (0.12-126.04, MEDIAN 3.92) (P > 0.05). CONCLUSION: OUR RESULTS DEMONSTRATE THAT ABERRANT METHYLATION OF DDIT3 OCCURS IN CML FREQUENTLY. 2010 10 4077 26 MATERNAL INFLAMMATION INDUCES SPATIAL LEARNING AND MEMORY IMPAIRMENT IN THE F1 AND F2 GENERATIONS OF MICE VIA SEX-SPECIFIC EPIGENETIC MECHANISMS. MOUNTING EVIDENCE INDICATES THAT HISTONE MODIFICATIONS ARE INVOLVED IN AGING-ASSOCIATED COGNITIVE DECLINE (AACD) AND CAN BE TRANSMITTED TO OFFSPRING OVER MULTIPLE GENERATIONS UNDER CONDITIONS OF STRESS. HERE, WE INVESTIGATED THE EFFECTS OF MATERNAL SUB-CHRONIC INFLAMMATION CAUSED BY LIPOPOLYSACCHARIDE (LPS) ON AACD AND HISTONE MODIFICATIONS IN THE F1 AND F2 GENERATIONS OF EXPERIMENTAL MICE AS WELL AS THE POTENTIAL SEX SPECIFICITY OF INTERGENERATIONAL EFFECTS. IN BRIEF, F0-GENERATION CD-1 DAMS WERE EXPOSED TO LPS (50 MICROG/KG) OR SALINE (CON) DURING LATE PREGNANCY. SUBSEQUENTLY, F1 MALES AND FEMALES (AT 2 MONTHS-OF-AGE) FROM THE LPS TREATMENT GROUP WERE MATED WITH NON-LITTERMATES FROM THE LPS GROUP OR WILD-TYPE MICE TO PRODUCE F2 GENERATIONS OF PARENTAL- (F2-LPS(2)), PATERNAL- (F2M-LPS(1)) AND MATERNAL-ORIGIN (F2F-LPS(1)) MICE. THEN, CON-F1 MALES AND FEMALES WERE MATED WITH WILD-TYPE MICE TO GENERATE F2 GENERATIONS OF PATERNAL- (F2M-CON(1)) AND MATERNAL-ORIGIN (F2F-CON(1)). NEXT, WE EVALUATED THE COGNITIVE ABILITY AND LEVELS OF HIPPOCAMPAL H4K12AC AND H3K9ME3 IN THE F1 AND F2 OFFSPRING AT 3- AND 13 MONTHS-OF-AGE. OVERALL, F1 MALE AND FEMALE LPS GROUPS PRESENTED WITH ELEVATED CORTICOSTERONE (P < 0.001, P = 0.036, P = 0.025, 0.012, RESPECTIVELY) AND CYTOKINE RESPONSES, POORER COGNITIVE PERFORMANCE (ALL P < 0.05) AND H3K9 HYPERMETHYLATION AND H4K12 HYPOACETYLATION IN THE DORSAL HIPPOCAMPUS (ALL P < 0.05); THESE ISSUES WERE CARRIED OVER TO THE F2 GENERATION VIA THE PARENTS, PREDOMINANTLY IN THE PATERNAL LINEAGE. MOREOVER, THE LEVELS OF H3K9ME3 AND H4K12AC WERE SIGNIFICANT CORRELATED WITH COGNITIVE PERFORMANCE (ALL P < 0.05), REGARDLESS OF WHETHER INFLAMMATORY INSULTS HAD BEEN INCURRED DIRECTLY OR INDIRECTLY. THESE FINDINGS INDICATED THAT GESTATIONAL INFLAMMATORY INSULTS IN THE F0 GENERATION ACCELERATED AACD IN THE F2 GENERATION, ALONG WITH H3K9 HYPERMETHYLATION AND H4K12 HYPOACETYLATION IN THE HIPPOCAMPUS, AND THAT THESE ISSUES WERE DERIVED FROM THE F1 PARENTS, ESPECIALLY FROM THE F1 FATHERS. 2022 11 3302 40 HIGH-FAT, SUCROSE AND SALT-RICH DIET DURING RAT SPERMATOGENESIS LEAD TO THE DEVELOPMENT OF CHRONIC KIDNEY DISEASE IN THE FEMALE OFFSPRING OF THE F2 GENERATION. EFFECTS OF FEEDING MALE RATS DURING SPERMATOGENESIS A HIGH-FAT, HIGH-SUCROSE AND HIGH-SALT DIET (HFSSD) OVER TWO GENERATIONS (F0 AND F1) ON RENAL OUTCOMES ARE UNKNOWN. MALE F0 AND F1 RATS WERE FED EITHER CONTROL DIET (F0CD+F1CD) OR HFSSD (F0HD+F1HD). THE OUTCOMES WERE GLOMERULAR FILTRATION RATE AND URINARY ALBUMIN EXCRETION IN F1 AND F2 OFFSPRING. IF BOTH OUTCOMES WERE ALTERED A MORPHOLOGICAL AND MOLECULAR ASSESSMENT WAS DONE. F2 OFFSPRING OF BOTH SEXES HAD A DECREASED GFR. HOWEVER, INCREASED URINARY ALBUMIN EXCRETION WAS ONLY OBSERVED IN FEMALE F2 F0HD+F1HD OFFSPRING COMPARED WITH CONTROLS. F0HD+F1HD FEMALE F2 OFFSPRING DEVELOPED GLOMERULOSCLEROSIS (+31%; P < .01) AND INCREASED RENAL INTERSTITIAL FIBROSIS (+52%; P < .05). RNA SEQUENCING FOLLOWED BY QRT-PCR VALIDATION SHOWED THAT FOUR GENES (ENPP6, TMEM144, CD300LF, AND ACTR3B) WERE DIFFERENTIALLY REGULATED IN THE KIDNEYS OF FEMALE F2 OFFSPRING. LNCRNA XR-146683.1 EXPRESSION DECREASED IN FEMALE F0HD+F1HD F2 OFFSPRING AND ITS EXPRESSION WAS (R = 0.44, P = .027) CORRELATED WITH THE EXPRESSION OF TMEM144. METHYLATION OF CPG ISLANDS IN THE PROMOTER REGION OF THE CD300LF GENE WAS INCREASED (P = .001) IN FEMALE F2 F0HD+F1HD OFFSPRING COMPARED TO CONTROLS. PROMOTER CPG ISLAND METHYLATION RATE OF CD300LF WAS INVERSELY CORRELATED WITH CD300LF MRNA EXPRESSION IN F2 FEMALE OFFSPRING (R = -0.483, P = .012). CD300LF MRNA EXPRESSION WAS INVERSELY CORRELATED WITH THE URINARY ALBUMIN-TO-CREATININE RATIO IN FEMALE F2 OFFSPRING (R = -0.588, P = .005). PATERNAL PRE-CONCEPTIONAL UNHEALTHY DIET GIVEN FOR TWO GENERATIONS PREDISPOSE FEMALE F2 OFFSPRING TO CHRONIC KIDNEY DISEASE DUE TO EPIGENETIC ALTERATIONS OF RENAL GENE EXPRESSION. PARTICULARLY, CD300LF GENE PROMOTOR METHYLATION WAS INVERSELY ASSOCIATED WITH CD300LF MRNA EXPRESSION AND CD300LF MRNA EXPRESSION ITSELF WAS INVERSELY ASSOCIATED WITH URINARY ALBUMIN EXCRETION IN F2 FEMALE OFFSPRING WHOSE FATHERS AND GRANDFATHERS GOT A PRE-CONCEPTIONAL UNHEALTHY DIET. 2022 12 403 32 ANALYSIS OF EPIGENETIC AGE ACCELERATION AND HEALTHY LONGEVITY AMONG OLDER US WOMEN. IMPORTANCE: ACCELERATED BIOLOGICAL AGING IS ASSOCIATED WITH DECREASED PHYSICAL CAPABILITY AND COGNITIVE FUNCTIONING, WHICH ARE ASSOCIATED WITH INCREASED RISK OF MORBIDITY AND MORTALITY. OBJECTIVE: WE INVESTIGATED ASSOCIATIONS BETWEEN EPIGENETIC AGE ACCELERATION (EAA), A BIOMARKER ASSOCIATED WITH AGING, AND HEALTHY LONGEVITY AMONG OLDER WOMEN. DESIGN, SETTING, AND PARTICIPANTS: THIS COHORT STUDY WAS A SECONDARY ANALYSIS OF PARTICIPANTS IN THE WOMEN'S HEALTH INITIATIVE (WHI) WHO WERE ELIGIBLE TO SURVIVE TO AGE 90 YEARS BY SEPTEMBER 30, 2020. PARTICIPANTS WERE LOCATED IN MULTIPLE CENTERS. THIS STUDY WAS RESTRICTED TO WOMEN WITH GENOME-WIDE DNA METHYLATION DATA, GENERATED FROM BASELINE BLOOD SAMPLES WITHIN 3 WHI ANCILLARY STUDIES. MEDIAN (IQR) FOLLOW-UP TIMES FROM BASELINE WERE 21.6 (19.6-22.9) YEARS AND 21.4 (19.8-22.7) YEARS FOR WOMEN WHO SURVIVED TO AGE 90 YEARS WITH AND WITHOUT INTACT MOBILITY, RESPECTIVELY, AND 13.2 (8.8-16.7) FOR WOMEN WHO DID NOT SURVIVE TO AGE 90 YEARS. DATA WERE ANALYZED FROM DECEMBER 2020 TO JULY 2021. EXPOSURES: EAA WAS ESTIMATED USING 4 ESTABLISHED "CLOCKS": HORVATH PANTISSUE, HANNUM, PHENO, AND GRIM. MAIN OUTCOMES AND MEASURES: USING MULTINOMIAL LOGISTIC REGRESSION, ODDS RATIOS (ORS) AND 95% CIS WERE ESTIMATED FOR 3 HEALTHY LONGEVITY OUTCOMES FOR EACH CLOCK: SURVIVAL TO AGE 90 YEARS WITH INTACT MOBILITY, SURVIVAL TO AGE 90 YEARS WITHOUT INTACT MOBILITY, AND NO SURVIVAL TO AGE 90 YEARS. RESULTS: AMONG 1813 WOMEN, THERE WERE 464 WOMEN (MEAN [SD] AGE AT BASELINE, 71.6 [3.5] YEARS) WHO SURVIVED TO AGE 90 YEARS WITH INTACT MOBILITY AND COGNITIVE FUNCTIONING, 420 WOMEN (MEAN [SD] AGE AT BASELINE, 71.3 [3.2] YEARS) WHO SURVIVED TO AGE 90 YEARS WITHOUT INTACT MOBILITY AND COGNITIVE FUNCTIONING, AND 929 WOMEN (MEAN [SD] AGE AT BASELINE, 70.2 [3.4] YEARS) WHO DID NOT SURVIVE TO AGE 90 YEARS. WOMEN WHO SURVIVED TO AGE 90 YEARS WITH INTACT MOBILITY AND COGNITIVE FUNCTION WERE HEALTHIER AT BASELINE COMPARED WITH WOMEN WHO SURVIVED WITHOUT THOSE OUTCOMES OR WHO DID NOT SURVIVE TO AGE 90 YEARS (EG, 143 WOMEN [30.8%] VS 101 WOMEN [24.0%] AND 202 WOMEN [21.7%] WITH 0 CHRONIC CONDITIONS). THE ODDS OF SURVIVING TO AGE 90 YEARS WITH INTACT MOBILITY WERE LOWER FOR EVERY 1 SD INCREASE IN EAA COMPARED WITH THOSE WHO DID NOT SURVIVE TO AGE 90 YEARS AS MEASURED BY AGEACCELHORVATH (OR, 0.82; 95% CI, 0.69-0.96; P = .01), AGEACCELHANNUM (OR, 0.67; 95% CI, 0.56-0.80; P < .001), AGEACCELPHENO (OR, 0.60; 95% CI, 0.51-0.72; P < .001), AND AGEACCELGRIM (OR, 0.68; 95% CI, 0.55-0.84; P < .001). ORS WERE SIMILAR FOR WOMEN WHO SURVIVED TO AGE 90 YEARS WITH INTACT MOBILITY AND COGNITIVE FUNCTION (EG, AGEACCELHORVATH: OR PER 1 SD INCREASE IN EAA, 0.83; 95% CI, 0.71-0.98; P = .03) COMPARED WITH WOMEN WHO DID NOT SURVIVE TO AGE 90 YEARS. CONCLUSIONS AND RELEVANCE: THESE FINDINGS SUGGEST THAT EAA MAY BE A VALID BIOMARKER ASSOCIATED WITH HEALTHY LONGEVITY AMONG OLDER WOMEN AND MAY BE USED FOR RISK STRATIFICATION AND RISK ESTIMATION OF FUTURE FUNCTIONAL AND COGNITIVE AGING. OUTCOMES SUGGEST THAT FUTURE STUDIES MAY FOCUS ON THE POTENTIAL FOR PUBLIC HEALTH INTERVENTIONS TO COUNTERACT EAA AND ITS ASSOCIATION WITH POOR HEALTH OUTCOMES TO LOWER DISEASE BURDEN WHILE INCREASING LONGEVITY. 2022 13 86 33 A PATERNAL HYPERCALORIC DIET AFFECTS THE METABOLISM AND FERTILITY OF F1 AND F2 WISTAR RAT GENERATIONS. INCREASED FAT AND CARBOHYDRATE INTAKES BASED ON THE WESTERN DIET ARE IMPORTANT LIFESTYLE MODIFICATIONS THAT LEAD TO HYPERCALORIC INPUTS, OBESITY, AND MALE FERTILITY NEGATIVE EFFECTS. EPIGENETIC TRANSMISSION MAY ALSO PREDISPOSE DESCENDED GENERATIONS TO CHRONIC DISEASES, SUCH AS OBESITY, TYPE 2 DIABETES, BEHAVIORAL, AND REPRODUCTIVE DISORDERS. THE PRESENT STUDY SOUGHT TO EVALUATE THE INFLUENCE OF A HIGH-FAT-HIGH-SUGAR (HFHS) DIET SUPPLIED TO WISTAR RATS FROM 25 TO 90 DAYS OF LIFE ON REPRODUCTIVE AND METABOLIC PARAMETERS IN MALE GENERATIONS F0, F1, AND F2. THE STANDARD GROUP RECEIVED THE NORMOCALORIC - NUVILAB QUIMTIA(R) -3.86 KCAL/KG. THE HYPERCALORIC DIET (HD) GROUP RECEIVED THE HFHS DIET - PRAGSOLUCOES(R) -4.77 KCAL/KG. BODY WEIGHT, ADIPOSITY, F1 AND F2 PREPUBERTAL AGE EVALUATIONS, ORAL GLUCOSE TOLERANCE TEST, INSULIN TOLERANCE TEST, ORGAN WEIGHTS, SPERM COUNT AND MORPHOLOGY ASSESSMENTS, AND HISTOMETRIC TESTICULAR ANALYSES WERE PERFORMED. THE HFHS DIET PROMOTED DYSLIPIDEMIA, HIGHER ADIPOSITY, LOWER RELATIVE ORGAN WEIGHTS, AND HIGHER MEAN KIDNEY WEIGHT, DECREASED MEAN TESTICLE AND PARENCHYMA WEIGHTS AND LOWER HEIGHT OF SEMINIFEROUS EPITHELIUM (HE) FOR THE F0 GENERATION. F1 AND F2 OFFSPRING OF HD GROUP DISPLAYED EARLY PREPREPUBERTAL DEVELOPMENT, ALTHOUGH DID NOT ALTER THE METABOLIC PARAMETERS. DECREASED HE AND TUBULAR TESTICULAR COMPARTMENT VOLUMETRIC DENSITY AND INCREASED INTERTUBULAR TESTICULAR COMPARTMENT VOLUMETRIC DENSITY AND VOLUME IN THE F1 GENERATION OF HD GROUP WERE OBSERVED. ALTERATIONS IN HISTOMETRY OF INTERTUBULAR TESTICULAR COMPARTMENT WERE ALSO NOTED. IT IS CONCLUDED THAT THE HFHS EXPERIMENTAL MODEL ALTERED ONLY PATERNAL METABOLIC PARAMETERS. HOWEVER, REPRODUCTIVE PARAMETERS OF THE THREE GENERATIONS WERE AFFECTED. 2020 14 3785 33 INTERGENERATIONAL EFFECTS OF PRE-PREGNANCY CHRONIC LIPOPOLYSACCHARIDE FROM PORPHYROMONAS GINGIVALIS ON THE LEARNING, MEMORY AND SEIZURE SUSCEPTIBILITY OF OFFSPRING. OBJECTIVE: THE AIM OF THIS STUDY WAS TO INVESTIGATE THE EFFECTS OF PRE-PREGNANCY CHRONIC EXPOSURE TO PORPHYROMONAS GINGIVALIS LPS (PG LPS) ON THE LEARNING, MEMORY, AND SEIZURE SUSCEPTIBILITY OF THE OFFSPRING. DESIGN: TO ACHIEVE PERIODONTITIS, PG LPS (5 MUG/KG) WAS INJECTED INTO THE GINGIVAL OF FIVE FEMALE RATS EVERY 48 H FOR THREE WEEKS. FIVE CONTROL FEMALE RATS RECEIVED SALINE (0.9 %) AND FIVE FEMALE WERE KEPT INTACT. THE CONCENTRATIONS OF TNF-ALPHA AND IL-6 WERE MEASURED IN THE BLOOD SAMPLES. ONE WEEK AFTER THE FINAL INJECTION, FEMALES WERE MATED WITH INTACT MALES. FOLLOWING BIRTH AND WEANING, TWO MALE AND TWO FEMALE OFFSPRING WERE RANDOMLY SELECTED FROM EACH MOTHER, AND NEW GROUPS OF MALE AND FEMALE OFFSPRING WERE DEFINED FOR BEHAVIORAL ASSESSMENTS. MORRIS WATER MAZE WAS USED TO EVALUATE SPATIAL MEMORY, SHUTTLE BOX WAS USED TO INVESTIGATE AVOIDANCE MEMORY AND A PENTYLENETETRAZOLE-INDUCED SEIZURE WAS USED TO EVALUATE SEIZURE SUSCEPTIBILITY IN THE OFFSPRING. RESULTS: SPATIAL LEARNING AND AVOIDANCE MEMORY SIGNIFICANTLY DECREASED IN BOTH MALE AND FEMALE OFFSPRING OF PG LPS-EXPOSED FEMALE RATS, COMPARED TO THE CONTROL OFFSPRING. LATENCY TO REACH SEIZURE STAGES 1 AND 2 SIGNIFICANTLY INCREASED IN THE MALE OFFSPRING, BUT NOT THE FEMALE OFFSPRING OF PG LPS-EXPOSED FEMALE, COMPARED TO THE CONTROL OFFSPRING. HOWEVER, NO SIGNIFICANT DIFFERENCE WAS FOUND IN LATENCY TO REACH STAGES 3-5. CONCLUSION: PRE-PREGNANCY EXPOSURE TO PG LPS COULD AFFECT SOME BEHAVIORAL FUNCTIONS IN BOTH MALE AND FEMALE OFFSPRING INTERGENERATIONALLY. 2021 15 3083 33 GENOME-WIDE SCREEN OF DNA METHYLATION CHANGES INDUCED BY LOW DOSE X-RAY RADIATION IN MICE. EPIGENETIC MECHANISMS PLAY A KEY ROLE IN NON-TARGETED EFFECTS OF RADIATION. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE GLOBAL HYPOMETHYLATION AND PROMOTER HYPERMETHYLATION OF PARTICULAR GENES INDUCED BY LOW DOSE RADIATION (LDR). THIRTY MALE BALB/C MICE WERE DIVIDED INTO 3 GROUPS: CONTROL, ACUTELY EXPOSED (0.5 GY X-RAYS), AND CHRONIC EXPOSURE FOR 10 DAYS (0.05GY/DX10D). HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) AND MEDIP-QUANTITATIVE POLYMERASE CHAIN REACTION (QPCR) WERE USED TO STUDY METHYLATION PROFILES. DNMT1 AND MBD2 EXPRESSION WAS DETERMINED BY QPCR AND WESTERN BLOT ASSAYS. METHYLATION AND EXPRESSION OF RAD23B AND DDIT3 WERE DETERMINED BY BISULFATE SEQUENCING PRIMERS (BSP) AND QPCR, RESPECTIVELY. THE RESULTS SHOW THAT LDR INDUCED GENOMIC HYPOMETHYLATION IN BLOOD 2 H POSTIRRADITAION, BUT WAS NOT RETAINED AT 1-MONTH. DNMT1 AND MBD2 WERE DOWNREGULATED IN A TISSUE-SPECIFIC MANNER BUT DID NOT PERSIST. SPECIFIC HYPERMETHYLATION WAS OBSERVED FOR 811 REGIONS IN THE GROUP RECEIVING CHRONIC EXPOSURE, WHICH COVERED ALMOST ALL KEY BIOLOGICAL PROCESSES AS INDICATED BY GO AND KEGG PATHWAY ANALYSIS. EIGHT HYPERMETHYLATED GENES (RAD23B, TDG, CCND1, DDIT3, LLGL1, RASL11A, TBX2, SCL6A15) WERE VERIFIED BY MEDIP-QPCR. AMONG THEM, RAD23B AND DDIT3 GENE DISPLAYED TISSUE-SPECIFIC METHYLATION AND DOWNREGULATION, WHICH PERSISTED FOR 1-MONTH POSTIRRADIATION. THUS, LDR INDUCED GLOBAL HYPOMETHYLATION AND TISSUE-SPECIFIC PROMOTER HYPERMETHYLATION OF PARTICULAR GENES. PROMOTER HYPERMETHYLATION, RATHER THAN GLOBAL HYPOMETHYLATION, WAS RELATIVELY STABLE. DYSREGULATION OF METHYLATION MIGHT BE CORRELATED WITH DOWN-REGULATION OF DNMT1 AND MBD2, BUT MUCH BETTER UNDERSTANDING THE MOLECULAR MECHANISMS INVOLVED IN THIS PROCESS WILL REQUIRE FURTHER STUDY. 2014 16 4504 24 MOTHER'S PRE-PREGNANCY BMI AND PLACENTAL CANDIDATE MIRNAS: FINDINGS FROM THE ENVIRONAGE BIRTH COHORT. THERE IS INCREASING EVIDENCE THAT THE PREDISPOSITION FOR DEVELOPMENT OF CHRONIC DISEASES ARISES AT THE EARLIEST TIMES OF LIFE. IN THIS CONTEXT, MATERNAL PRE-PREGNANCY WEIGHT MIGHT MODIFY FETAL METABOLISM AND THE CHILD'S PREDISPOSITION TO DEVELOP DISEASE LATER IN LIFE. THE AIM OF THIS STUDY IS TO INVESTIGATE THE ASSOCIATION BETWEEN MATERNAL PRE-PREGNANCY BODY MASS INDEX (BMI) AND MIRNA ALTERATIONS IN PLACENTAL TISSUE AT BIRTH. IN 211 MOTHER-NEWBORN PAIRS FROM THE ENVIRONAGE BIRTH COHORT, WE ASSESSED PLACENTAL EXPRESSION OF SEVEN MIRNAS IMPORTANT IN CRUCIAL CELLULAR PROCESSES IMPLICATED IN ADIPOGENESIS AND/OR OBESITY. MULTIPLE LINEAR REGRESSION MODELS WERE USED TO ADDRESS THE ASSOCIATIONS BETWEEN PRE-PREGNANCY BMI AND PLACENTAL CANDIDATE MIRNA EXPRESSION. MATERNAL PRE-PREGNANCY BMI AVERAGED (+/-SD) 23.9 (+/-4.1) KG/M(2). IN NEWBORN GIRLS (NOT IN BOYS) PLACENTAL MIR-20A, MIR-34A AND MIR-222 EXPRESSION WAS LOWER WITH HIGHER MATERNAL PRE-PREGNANCY BMI. IN ADDITION, THE ASSOCIATION BETWEEN MATERNAL PRE-PREGNANCY BMI AND PLACENTAL EXPRESSION OF THESE MIRNAS IN GIRLS WAS MODIFIED BY GESTATIONAL WEIGHT GAIN. THE LOWER EXPRESSION OF THESE MIRNAS IN PLACENTA IN ASSOCIATION WITH PRE-PREGNANCY BMI, WAS ONLY EVIDENT IN MOTHERS WITH LOW WEIGHT GAIN (<14 KG). THE PLACENTAL EXPRESSION OF MIR-20A, MIR-34A, MIR-146A, MIR-210 AND MIR-222 MAY PROVIDE A SEX-SPECIFIC BASIS FOR EPIGENETIC EFFECTS OF PRE-PREGNANCY BMI. 2017 17 138 28 ABERRANT DNA METHYLATION AT HOXA4 AND HOXA5 GENES ARE ASSOCIATED WITH RESISTANCE TO IMATINIB MESYLATE AMONG CHRONIC MYELOID LEUKEMIA PATIENTS. BACKGROUND: IMATINIB MESYLATE IS A MOLECULARLY TARGETED TYROSINE KINASE INHIBITOR DRUG. IT IS EFFECTIVELY USED IN THE TREATMENT OF CHRONIC MYELOID LEUKEMIA (CML) PATIENTS. HOWEVER, DEVELOPMENT OF RESISTANCE TO IMATINIB MESYLATE AS A RESULT OF BCR-ABL DEPENDENT AND BCR-ABL INDEPENDENT MECHANISMS HAS EMERGED AS A DAUNTING PROBLEM IN THE MANAGEMENT OF CML PATIENTS. BETWEEN THESE MECHANISMS, BCR-ABL INDEPENDENT MECHANISMS ARE STILL NOT ROBUSTLY UNDERSTOOD. AIM: TO INVESTIGATE THE CORRELATION OF HOXA4 AND HOXA5 PROMOTER DNA HYPERMETHYLATION WITH IMATINIB RESISTANCE AMONG CML PATIENTS. METHODS AND RESULTS: SAMPLES FROM 175 PHILADELPHIA POSITIVE CML PATIENTS (83 GOOD RESPONSE AND 92 BCR-ABL NON-MUTATED IMATINIB RESISTANT PATIENTS) WERE SUBJECTED TO METHYLATION SPECIFIC HIGH RESOLUTION MELT ANALYSIS FOR METHYLATION LEVELS QUANTIFICATION OF THE HOXA4 AND HOXA5 PROMOTER REGIONS. RECEIVER OPERATING CHARACTERISTIC CURVE ANALYSIS WAS DONE TO ELUCIDATE THE OPTIMAL METHYLATION CUT-OFF POINT FOLLOWED BY MULTIPLE LOGISTIC REGRESSION ANALYSIS. LOG-RANK ANALYSIS WAS DONE TO MEASURE THE OVERALL SURVIVAL DIFFERENCE BETWEEN CML GROUPS. THE OPTIMAL METHYLATION CUT-OFF POINT WAS FOUND TO BE AT 62.5% FOR BOTH HOXA4 AND HOXA5. CHRONIC MYELOID LEUKEMIA PATIENTS WITH >/=63% HOXA4 AND HOXA5 METHYLATION LEVEL WERE SHOWN TO HAVE 3.78 AND 3.95 TIMES THE ODDS, RESPECTIVELY, TO ACQUIRE RESISTANCE TO IMATINIB. HOWEVER, OVERALL SURVIVAL OF CML PATIENTS THAT HAVE /= 63% METHYLATION LEVELS OF HOXA4 AND HOXA5 GENES WERE FOUND TO BE NOT SIGNIFICANT (P-VALUE = 0.126 FOR HOXA4; P-VALUE = 0.217 FOR HOXA5). CONCLUSION: HYPERMETHYLATION OF THE HOXA4 AND HOXA5 PROMOTER IS CORRELATED WITH IMATINIB RESISTANCE AND WITH FURTHER INVESTIGATION, IT COULD BE A POTENTIAL EPIGENETIC BIOMARKER IN SUPPLEMENT TO THE BCR-ABL GENE MUTATION IN PREDICTING IMATINIB TREATMENT RESPONSE AMONG CML PATIENTS BUT COULD NOT BE CONSIDERED AS A PROGNOSTIC MARKER. 2018 18 4021 29 LOWERED DNA METHYLTRANSFERASE (DNMT-3B) MRNA EXPRESSION IS ASSOCIATED WITH GENOMIC DNA HYPERMETHYLATION IN PATIENTS WITH CHRONIC ALCOHOLISM. DNA METHYLTRANSFERASES (DNMTS) ARE INVOLVED WITHIN THE EPIGENETIC CONTROL OF DNA METHYLATION PROCESSES. RECENTLY, IT HAS BEEN SHOWN THAT THE GENOMIC DNA METHYLATION IN PATIENTS WITH ALCOHOLISM IS INCREASED. IN THE PRESENT CONTROLLED STUDY WE OBSERVED A SIGNIFICANT DECREASE OF MRNA EXPRESSION OF DNMT-3A AND DNMT-3B WHEN COMPARING ALCOHOLIC PATIENTS (N = 59) WITH HEALTHY CONTROLS (N = 66): DNMT-3A (T = -2.38, P = 0.019), DNMT-3B (T = -2.65, P = 0.008). NO SIGNIFICANT DIFFERENCES WERE SEEN FOR DNMT-1 AND MBD-2 (METHYL-CPG-BINDING-DOMAIN PROTEIN 2) EXPRESSION. ADDITIONALLY, WE OBSERVED A SIGNIFICANT NEGATIVE CORRELATION BETWEEN DNMT-3B EXPRESSION AND THE BLOOD ALCOHOL CONCENTRATION (R = -0.45, P = 0.003) WHICH MIGHT EXPLAIN THE DECREASE OF DNMT-3B MRNA EXPRESSION IN ALCOHOLIC PATIENTS. USING A MULTIVARIATE MODEL WE OBSERVED THAT THE INCREASE (10%) OF GENOMIC DNA METHYLATION IN PATIENTS WITH ALCOHOLISM WAS SIGNIFICANTLY ASSOCIATED WITH THEIR LOWERED DNMT-3B MRNA EXPRESSION (MULTIPLE LINEAR REGRESSION, P = 0.014). 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 IN THESE PATIENTS. 2006 19 1849 23 EIGHT WEEKS OF PHYSICAL TRAINING DECREASES 2 YEARS OF DNA METHYLATION AGE OF SEDENTARY WOMEN. PURPOSE: THE ACCELERATION OF EPIGENETIC AGE IS A PREDICTOR OF MORTALITY AND CONTRIBUTES TO THE INCREASE IN CHRONIC DISEASES. ADHERENCE TO A HEALTHY LIFESTYLE IS A STRATEGY TO REDUCE EPIGENETIC AGE. THE PRESENT STUDY AIMED TO DETERMINE WHETHER EIGHT WEEKS OF COMBINED (AEROBIC AND STRENGTH) TRAINING (CT) CAN INFLUENCE THE EPIGENETIC AGE OF WOMEN BETWEEN 50 AND 70 YEARS OLD AND THE DIFFERENCES IN SITES AND METHYLATED REGIONS. METHODS: EIGHTEEN WOMEN (AAR(LOW): LOWER AGE ACCELERATION RESIDUAL, N = 10; AAR(HIGH): HIGHER AGE ACCELERATION RESIDUAL, N = 8) PARTICIPATED IN A COMBINED EXERCISE TRAINING PROGRAM (60 MINUTES, 3X A WEEK) FOR EIGHT WEEKS. DNA WAS EXTRACTED FROM WHOLE BLOOD USING THE SALTING OUT TECHNIQUE. DNA METHYLATION WAS PERFORMED USING THE ARRAY TECHNIQUE (ILLUMINA'S INFINIUM METHYLATION BEADCHIP 850K). WE USED THE DNA METHYLATION AGE CALCULATOR PLATFORM TO CALCULATE THE BIOLOGICAL EPIGENETIC AGE. TWO-WAY ANOVA FOLLOWED BY FISHER LSD POSTHOC WAS APPLIED, ADOPTING P < .05. RESULTS: AFTER EIGHT WEEKS OF CT, THERE WERE NO CHANGES TO THE EPIGENETIC AGE ACCELERATION FOR THE AAR(LOW) GROUP (PRE: -2.3 +/- 3.2 TO POST: -2.3 +/- 3.6). HOWEVER, THE AAR(HIGH) GROUP SIGNIFICANTLY DECREASED THE AGE ACCELERATION (PRE: 3.6 +/- 2.6 TO POST: 2.2 +/- 2.7) (GROUP EFFECT, P = .01; TIME EFFECT, P = .31; GROUP VS. TIME EFFECT, P = .005). CONCLUSION: CT FOR EIGHT WEEKS BENEFITS THE EPIGENETIC CLOCK OF WOMEN WITH THE MOST ACCELERATED AGE. 2023 20 3410 37 HOXA5 UNDERGOES DYNAMIC DNA METHYLATION AND TRANSCRIPTIONAL REPRESSION IN THE ADIPOSE TISSUE OF MICE EXPOSED TO HIGH-FAT DIET. BACKGROUND/OBJECTIVES: THE GENOMIC BASES OF THE ADIPOSE TISSUE ABNORMALITIES INDUCED BY CHRONIC POSITIVE CALORIE EXCESS HAVE BEEN ONLY PARTIALLY ELUCIDATED. WE ADOPTED A GENOME-WIDE APPROACH TO DIRECTLY TEST WHETHER LONG-TERM HIGH-FAT DIET (HFD) EXPOSURE AFFECTS THE DNA METHYLATION PROFILE OF THE MOUSE ADIPOSE TISSUE AND TO IDENTIFY THE FUNCTIONAL CONSEQUENCES OF THESE CHANGES. SUBJECTS/METHODS: WE HAVE USED EPIDIDYMAL FAT OF MICE FED EITHER HIGH-FAT (HFD) OR REGULAR CHOW (STD) DIET FOR 5 MONTHS AND PERFORMED GENOME-WIDE DNA METHYLATION ANALYSES BY METHYLATED DNA IMMUNOPRECIPITATION SEQUENCING (MEDIP-SEQ). MOUSE HOMEOBOX (HOX) GENE DNA METHYLATION PCR, RT-QPCR AND BISULPHITE SEQUENCING ANALYSES WERE THEN PERFORMED. RESULTS: MICE FED THE HFD PROGRESSIVELY EXPANDED THEIR ADIPOSE MASS ACCOMPANIED BY A SIGNIFICANT DECREASE IN GLUCOSE TOLERANCE (P<0.001) AND INSULIN SENSITIVITY (P<0.05). MEDIP-SEQ DATA ANALYSIS REVEALED A UNIFORM DISTRIBUTION OF DIFFERENTIALLY METHYLATED REGIONS (DMR) THROUGH THE ENTIRE ADIPOCYTE GENOME, WITH A HIGHER NUMBER OF HYPERMETHYLATED REGIONS IN HFD MICE (P<0.005). THIS DIFFERENT METHYLATION PROFILE WAS ACCOMPANIED BY INCREASED EXPRESSION OF THE DNMT3A DNA METHYLTRANSFERASE (DNMT; P<0.05) AND THE METHYL-CPG-BINDING DOMAIN PROTEIN MBD3 (P<0.05) GENES IN HFD MICE. GENE ONTOLOGY ANALYSIS REVEALED THAT, IN THE HFD-TREATED MICE, THE HOX FAMILY OF DEVELOPMENT GENES WAS HIGHLY ENRICHED IN DIFFERENTIALLY METHYLATED GENES (P=0.008). TO VALIDATE THIS FINDING, HOXA5, WHICH IS IMPLICATED IN FAT TISSUE DIFFERENTIATION AND REMODELING, HAS BEEN SELECTED AND ANALYZED BY BISULPHITE SEQUENCING, CONFIRMING HYPERMETHYLATION IN THE ADIPOSE TISSUE FROM THE HFD MICE. HOXA5 HYPERMETHYLATION WAS ASSOCIATED WITH DOWNREGULATION OF HOXA5 MRNA AND PROTEIN EXPRESSION. FEEDING ANIMALS PREVIOUSLY EXPOSED TO THE HFD WITH A STANDARD CHOW DIET FOR TWO FURTHER MONTHS IMPROVED THE METABOLIC PHENOTYPE OF THE ANIMALS, ACCOMPANIED BY RETURN OF HOXA5 METHYLATION AND EXPRESSION LEVELS (P<0.05) TO VALUES SIMILAR TO THOSE OF THE CONTROL MICE MAINTAINED UNDER STANDARD CHOW. CONCLUSIONS: HFD INDUCES ADIPOSE TISSUE ABNORMALITIES ACCOMPANIED BY EPIGENETIC CHANGES AT THE HOXA5 ADIPOSE TISSUE REMODELING GENE. 2016