1   161 140 ABHD4-REGULATING RNA PANEL: NOVEL BIOMARKERS IN ACUTE CORONARY SYNDROME DIAGNOSIS. BACKGROUND: ACUTE CORONARY SYNDROME (ACS) IS A MAJOR CAUSE OF DEATH ALL OVER THE WORLD. STEMI REPRESENTS A TYPE OF MYOCARDIAL INFARCTION WITH ACUTE ST ELEVATION. WE AIMED TO ASSESS THE PREDICTIVE POWER OF POTENTIAL RNA PANEL EXPRESSION IN ACUTE CORONARY SYNDROME. METHOD: WE USED IN SILICO DATA ANALYSIS TO RETRIEVE RNAS RELATED TO GLYCEROPHOSPHOLIPID METABOLISM DYSREGULATION AND SPECIFIC TO ACS THAT RESULTS IN THE SELECTION OF ALPHA/BETA HYDROLASE FOLD DOMAIN4 (ABHD4) MRNA AND ITS EPIGENETIC REGULATORS (FOXF1 ADJACENT NONCODING DEVELOPMENTAL REGULATORY RNA (FENDRR) LNCRNA, MIRNA-221, AND MIRNA-197). WE ASSESSED THE EXPRESSION OF THE SERUM RNA PANEL IN 68 PATIENTS WITH ACS, 21 PATIENTS WITH CHEST PAIN DUE TO NON-CARDIAC CAUSES, AND 21 HEALTHY VOLUNTEERS BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION. RESULTS: THE STUDY DATA SHOWED SIGNIFICANT DOWN REGULATION IN THE EXPRESSION OF THE SERUM LEVELS OF FENDRR LNCRNA AND MIRNA-221-3P BY 120-FOLD AND 22-FOLD IN UNSTABLE ANGINA (UA) IN COMPARISON WITH HEALTHY VOLUNTEERS, AND BY 8.6-FOLD AND 2-FOLD IN ST SEGMENT ELEVATION MYOCARDIAL INFARCTION (STEMI) PATIENTS VERSUS UA; CONCOMITANT UPREGULATION IN THE EXPRESSION OF ABHD4 MRNA AND MIRNA-197-5P BY 444-FOLD AND 10-FOLD IN UA COMPARED WITH HEALTHY VOLUNTEERS, AND BY 1.54-FOLD AND 4.5-FOLD IN STEMI VERSUS UNSTABLE ANGINA. PERFORMANCE CHARACTERISTICS ANALYSIS SHOWED THAT THE ABHD4-REGULATING RNA PANEL WERE POTENTIAL BIOMARKERS FOR PREDICTION OF ACS. MOREOVER, THERE WAS A SIGNIFICANT ASSOCIATION BETWEEN THE 2 MIRNAS AND ABHD4 MRNA AND THE REGULATING FENDRR LNCRNA. CONCLUSION: COLLECTIVELY, ABHD4 MRNA REGULATING RNA PANEL BASED ON PUTATIVE INTERACTIONS SEEMS TO BE NOVEL NON-INVASIVE BIOMARKERS THAT COULD DETECT ACS EARLY AND STRATIFY SEVERITY OF THE CONDITION THAT COULD IMPROVE HEALTH OUTCOME.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
2   487  33 ASPIRIN PROTECTS HUMAN CORONARY ARTERY ENDOTHELIAL CELLS AGAINST ATHEROGENIC ELECTRONEGATIVE LDL VIA AN EPIGENETIC MECHANISM: A NOVEL CYTOPROTECTIVE ROLE OF ASPIRIN IN ACUTE MYOCARDIAL INFARCTION. AIMS: L5 IS THE MOST NEGATIVELY CHARGED SUBFRACTION OF HUMAN LOW-DENSITY LIPOPROTEIN (LDL) AND IS THE ONLY SUBFRACTION OF LDL CAPABLE OF INDUCING APOPTOSIS IN CULTURED VASCULAR ENDOTHELIAL CELLS (ECS) BY INHIBITING FIBROBLAST GROWTH FACTOR-2 (FGF2) TRANSCRIPTION. WE EXAMINED WHETHER PLASMA L5 LEVELS ARE ELEVATED IN PATIENTS WITH ST-SEGMENT ELEVATION MYOCARDIAL INFARCTION (STEMI) AND WHETHER ASPIRIN PROVIDES EPIGENETIC PROTECTION OF HUMAN CORONARY ARTERY ECS (HCAECS) EXPOSED TO L5. METHODS AND RESULTS: PLASMA L5 LEVELS WERE COMPARED BETWEEN PATIENTS WITH STEMI (N = 10) AND CONTROL SUBJECTS WITH CHEST PAIN SYNDROME BUT A NORMAL CORONARY ARTERIOGRAM (N = 5). L5 WAS ISOLATED FROM THE PLASMA OF STEMI PATIENTS AND CONTROL SUBJECTS, AND APOPTOSIS, FGF2 EXPRESSION, AND FGF2 PROMOTER METHYLATION WERE EXAMINED IN HCAECS TREATED WITH L5 AND ASPIRIN. PLASMA L5 LEVELS WERE SIGNIFICANTLY HIGHER IN STEMI PATIENTS THAN IN CONTROL SUBJECTS (P < 0.001). TREATMENT OF HCAECS WITH L5 RESULTED IN REDUCED SURVIVAL AND FGF2 EXPRESSION AND INCREASED CPG METHYLATION OF THE FGF2 PROMOTER. CO-TREATMENT OF HCAECS WITH L5 AND A PHYSIOLOGICALLY RELEVANT, LOW CONCENTRATION OF ASPIRIN (0.2 MM) ATTENUATED THE ADVERSE EFFECTS OF L5 ON HCAEC SURVIVAL, FGF2 EXPRESSION, AND FGF2 PROMOTER METHYLATION. IN CONTRAST, HIGH CONCENTRATIONS OF ASPIRIN (>/=1.0 MM) ACCENTUATED THE EFFECTS OF L5. CONCLUSIONS: OUR RESULTS SHOW THAT L5 LEVELS ARE SIGNIFICANTLY INCREASED IN STEMI PATIENTS. FURTHERMORE, L5 IMPAIRS HCAEC FUNCTION THROUGH CPG METHYLATION OF THE FGF2 PROMOTER, WHICH IS SUPPRESSED IN THE PRESENCE OF LOW-CONCENTRATION ASPIRIN. OUR RESULTS PROVIDE EVIDENCE OF A NOVEL MECHANISM OF ASPIRIN IN THE PREVENTION OF MI.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
3  4601  35 NDRG2 MRNA LEVELS AND MIR-28-5P AND MIR-650 ACTIVITY IN CHRONIC LYMPHOCYTIC LEUKEMIA. BACKGROUND: NDRG2 IS IDENTIFIED AS A TUMOR SUPPRESSOR GENE IN MANY TUMORS, AND FUNCTIONS IN CELL PROLIFERATION, DIFFERENTIATION AND APOPTOSIS. RECENT DATA INDICATE THAT NDRG2 EXPRESSION IS UP-REGULATED BY TP53. MOREOVER, PROPOSED MECHANISMS OF NDRG2 INACTIVATION INCLUDE EPIGENETIC SILENCING OF THE NDRG2 PROMOTER AND DOWN-REGULATION BY MICRORNAS (MIRNAS). HOWEVER, FEW STUDIES HAVE EVER BEEN DONE ON THE ROLE OF NDRG2 AND THE NDRG2-REGULATING MIRNAS INTERFERENCE IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). METHODS: NDRG2 AND MICRORNAS MRNA LEVELS IN CLL SUBJECTS WERE ASSESSED BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QRT-PCR). THE DUAL-LUCIFERASE REPORTER ASSAY WAS PERFORMED TO DETERMINE NDRG2-RELATED MIRNAS. LOW EXPRESSION OF MATURE EXOGENOUS MIRNAS IN CLL CELLS WAS ESTABLISHED BY TRANSIENT TRANSFECTION. NDRG2 PROTEIN LEVELS IN CLL CELLS WERE DETECTED BY WESTERN BLOT. IN ADDITION, FLOW CYTOMETRY WAS CONDUCTED TO EXAMINE THE APOPTOSIS OF CLL CELLS. RESULTS: LOWER EXPRESSION OF NDRG2 WAS FOUND IN THE B-CELLS FROM 102 CLL PATIENTS COMPARED THE 40 NORMAL SUBJECTS (P < 0.001). PATIENTS WITH ADVANCED BINET STAGE (P = 0.001), HIGH LACTATE DEHYDROGENASE (LDH) LEVEL (P = 0.036), UN-MUTATED IMMUNOGLOBULIN HEAVY CHAIN VARIABLE REGION GENE (IGHV) (P = 0.004) AND THOSE WITH P53 ABERRATIONS (P < 0.001) HAD A MARKEDLY LOWER LEVELS OF NDRG2 MRNA. THIS DECREASE WAS ASSOCIATED WITH BRIEFER TIME-TO-TREATMENT (P = 0.001) AND POORER SURVIVAL (P < 0.001). HIGH EXPRESSION OF MIR-28-5P AND MIR-650 WAS ASSOCIATED WITH BINET B/C STAGE (P = 0.044) AND IGHV UN-MUTATED (P = 0.011), AS WELL AS BINET B/C STAGE (P = 0.013) AND P53 ABERRATIONS (P = 0.037), RESPECTIVELY. INHIBITION OF MIR-28-5P OR MIR-650 COULD INDUCE MORE APOPTOSIS IN CLL CELLS WITH GERMLINE TP53. CONCLUSIONS: NDRG2 MRNA LEVELS MIGHT BE A USEFUL PROGNOSTIC VARIABLE FOR PATIENTS OF CLL AND UP-REGULATING NDRG2 TRANSCRIPTION MAY BE A THERAPY APPROACH IN CLL WITHOUT P53 ABERRATIONS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
4  6105  27 THE EMERGING ROLE OF LNCRNA FENDRR IN MULTIPLE CANCERS: A REVIEW. LONG NONCODING RNAS (LNCRNAS) ARE PROMINENT AS CRUCIAL REGULATORS OF TUMOR ESTABLISHMENT AND ARE REPEATEDLY DYSREGULATED IN MULTIPLE CANCERS. THEREFORE, LNCRNAS HAVE BEEN IDENTIFIED TO PLAY AN ESSENTIAL FUNCTION IN CARCINOGENESIS AND PROGRESSION OF CANCER AT GENETIC AND EPIGENETIC LEVELS. FENDRR (FETAL-LETHAL NONCODING DEVELOPMENTAL REGULATORY RNA) AS A LNCRNA IS A HALLMARK OF VARIOUS MALIGNANCIES. FENDRR IS CRUCIAL FOR MULTIPLE ORGANS' DEVELOPMENT, SUCH AS THE LUNG AND HEART. THE EFFECTS OF FENDRR UNDER SIGNALING PATHWAYS IN DIFFERENT CANCERS HAVE BEEN IDENTIFIED. IN ADDITION, IT HAS BEEN VERIFIED THAT FENDRR CAN AFFECT THE DEVELOPMENT AND PROGRESSION OF VARIOUS CANCERS. IN ADDITION, FENDRR EXPRESSION HAS BEEN ASSOCIATED WITH EPIGENETIC REGULATION OF TARGET GENES PARTICIPATING IN TUMOR IMMUNITY. FURTHERMORE, FENDRR DOWNREGULATION WAS OBSERVED IN VARIOUS TYPES OF CANCERS, INCLUDING COLORECTAL CANCER, GASTRIC CANCER, PANCREATIC CANCER, CHOLANGIOCARCINOMA, LIVER CANCER, GALLBLADDER CANCER, LUNG CANCER, BREAST CANCER, ENDOMETRIAL CANCER, PROSTATE CANCER, CHRONIC MYELOID LEUKEMIA, OSTEOSARCOMA, AND CUTANEOUS MALIGNANT MELANOMA CELLS. HERE, WE REVIEW THE BIOLOGICAL FUNCTIONS AND MOLECULAR MECHANISMS OF FENDRR IN SEVERAL CANCERS, AND WE WILL DISCUSS ITS POTENTIAL AS A CANCER BIOMARKER AND AS A PROBABLE OPTION FOR CANCER TREATMENT.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
5   779  26 CELL-FREE DNA PROMOTER HYPERMETHYLATION AS A DIAGNOSTIC MARKER FOR PANCREATIC DUCTAL ADENOCARCINOMA - AN EXTERNAL VALIDATION STUDY. BACKGROUND: WE RECENTLY IDENTIFIED A DIAGNOSTIC PREDICTION MODEL BASED ON PROMOTER HYPERMETHYLATION OF EIGHT SELECTED GENES IN PLASMA CELL-FREE (CF) DNA, WHICH SHOWED PROMISING RESULTS AS A DIAGNOSTIC BIOMARKER FOR PANCREATIC DUCTAL ADENOCARCINOMA (PDAC). THE AIM OF THE PRESENT STUDY WAS TO VALIDATE THIS BIOMARKER PROFILE IN AN EXTERNAL PATIENT COHORT AND EXAMINE ANY ADDITIONAL EFFECT OF SERUM CA 19-9. METHODS: PATIENTS WITH PDAC (N = 346, STAGE I-IV) AND CHRONIC PANCREATITIS (N = 25) WERE INCLUDED. METHYLATION-SPECIFIC PCR OF A 28-GENE PANEL WAS PERFORMED ON SERUM CFDNA SAMPLES. THE PREVIOUSLY DEVELOPED DIAGNOSTIC PREDICTION MODEL (AGE>65 YEARS, BMP3, RASSF1A, BNC1, MESTV2, TFPI2, APC, SFRP1 AND SFRP2) WAS VALIDATED ALONE AND IN COMBINATION WITH SERUM CA 19-9 IN THIS EXTERNAL PATIENT COHORT. RESULTS: PATIENTS WITH PDAC HAD A HIGHER NUMBER OF HYPERMETHYLATED GENES (MEAN 8.11, 95% CI 7.70-8.52) THAN PATIENTS WITH CHRONIC PANCREATITIS (MEAN 5.60, 95% CI 4.42-6.78, P = 0.011). VALIDATION OF THE DIAGNOSTIC PREDICTION MODEL YIELDED AN AUC OF 0.77 (95% CI 0.69-0.84). THE COMBINATION OF SERUM CA 19-9 AND OUR TEST HAD AN AUC OF 0.93 (95% CI 0.89-0.96) IN THE PRIMARY STUDY AND 0.85 (95% CI 0.79-0.91) IN THE VALIDATION STUDY. CONCLUSION: IN THIS VALIDATION STUDY, PDAC WAS ASSOCIATED WITH A HIGHER NUMBER OF HYPERMETHYLATED GENES IN SERUM CFDNA THAN CHRONIC PANCREATITIS. OUR DIAGNOSTIC TEST WAS SUPERIOR TO THE PREDICTIVE VALUE OF SERUM CA 19-9 ALONE IN BOTH THE PRIMARY AND THE VALIDATION STUDY. THE COMBINATION OF OUR TEST WITH CA 19-9 MAY SERVE AS A CLINICALLY USEFUL DIAGNOSTIC BIOMARKER FOR PDAC.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
6  6596  32 TUMOR-SUPPRESSIVE MIR-192-5P HAS PROGNOSTIC VALUE IN PANCREATIC DUCTAL ADENOCARCINOMA. PANCREATIC DUCTAL ADENOCARCINOMA (PDAC) IS CHARACTERIZED BY FAST TUMOR PROGRESSION AND DIAGNOSIS AT ADVANCED, INOPERABLE STAGES. PREVIOUS STUDIES COULD DEMONSTRATE AN INVOLVEMENT OF MIR-192-5P IN EPIGENETIC REGULATION OF VISCERAL CARCINOMAS. DUE TO CONTRADICTORY RESULTS, HOWEVER, THE CLINICAL UTILITY OF MIR-192-5P IN PDAC HAS YET TO BE DETERMINED. MIR-192-5P EXPRESSION WAS ANALYZED BY RT-QRT-PCR IN HUMAN PDAC AND BENIGN TISSUE (N = 78), BLOOD SERUM (N = 81) AND SERUM EXOSOMES (N = 74), AS WELL AS IN PDAC CELL LINES (N = 5), CHEMORESISTANT CELL CLONES (N = 2), AND PANCREATIC DUCT CELL LINE H6C7. ANALYSIS OF EMT-ASSOCIATED (EPITHELIAL-TO-MESENCHYMAL TRANSITION) PROTEINS WAS PERFORMED BY IMMUNOHISTOCHEMISTRY AND WESTERN BLOT. MIR-192-5P WAS DEREGULATED IN PDAC AS COMPARED TO HEALTHY CONTROLS (HCS), WITH DOWNREGULATION IN MACRODISSECTED TISSUE (P < 0.001) AND UPREGULATION IN BLOOD SERUM OF PDAC UICC (UNION FOR INTERNATIONAL CANCER CONTROL) STAGE IV (P = 0.016) AND SERUM EXOSOMES OF PDAC UICC STAGES II TO IV (P < 0.001). MIR-192-5P EXPRESSION IN TUMOR TISSUE WAS SIGNIFICANTLY LOWER AS COMPARED TO CORRESPONDING PERITUMORAL TISSUE (PDAC UICC STAGE II: P < 0.001; PDAC UICC STAGE III: P = 0.024), WHILE EMT MARKERS ZEB1 AND ZEB2 WERE MORE FREQUENTLY EXPRESSED IN TUMOR TISSUE AS COMPARED TO PERITUMORAL TISSUE, HCS, AND CHRONIC PANCREATITIS. TISSUE-DERIVED (AUC OF 0.86; P < 0.0001) AND EXOSOMAL (AUC OF 0.83; P = 0.0004) MIR-192-5P COULD DIFFERENTIATE BETWEEN PDAC AND HCS WITH GOOD ACCURACY. FURTHERMORE, HIGH EXPRESSION OF MIR-192-5P IN PDAC TISSUE OF CURATIVELY RESECTED PDAC PATIENTS CORRELATED WITH PROLONGED OVERALL AND RECURRENCE-FREE SURVIVAL IN MULTIVARIATE ANALYSIS. IN VITRO, MIR-192-5P WAS DOWNREGULATED IN GEMCITABINE-RESISTANT CELL CLONES OF ASPC-1 (P = 0.029). TRANSIENT TRANSFECTION OF MIA PACA-2 CELLS WITH MIR-192-5P MIMIC RESULTED IN DOWNREGULATION OF ZEB2. MIR-192-5P SEEMS TO POSSESS A TUMOR-SUPPRESSIVE ROLE AND HIGH POTENTIAL AS A DIAGNOSTIC AND PROGNOSTIC MARKER IN PDAC.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
7   659  32 BLOOD GLOBAL DNA METHYLATION IS DECREASED IN NON-SEVERE CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) PATIENTS. BACKGROUND: ALTERATIONS IN GLOBAL DNA METHYLATION HAVE BEEN ASSOCIATED WITH OXIDATIVE STRESS (OS). SINCE CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) IS CHARACTERIZED BY INCREASED OXIDATIVE STRESS WE AIMED TO EVALUATE THE LEVELS OF GLOBAL DNA METHYLATION IN THIS PATIENT GROUP. METHODS: WE ASSESSED METHYLCYTOSINE (MCYT) LEVELS IN DNA FROM BLOOD COLLECTED IN 43 COPD PATIENTS (29 WITH MILD AND 14 WITH MODERATE DISEASE) AND 43 AGE- AND SEX-MATCHED HEALTHY CONTROLS. RESULTS: DNA METHYLATION WAS SIGNIFICANTLY LOWER IN COPD PATIENTS VS. CONTROLS (4.20 +/- 0.18% MCYT VS. 4.29 +/- 0.18% MCYT, P = 0.02). FURTHERMORE, DNA METHYLATION IN COPD PATIENTS WITH MODERATE DISEASE WAS SIGNIFICANTLY LOWER THAN THAT IN PATIENTS WITH MILD DISEASE (4.14 +/- 0.15% MCYT VS. 4.23 +/- 0.19% MCYT, P < 0.05). UNIVARIATE LOGISTIC REGRESSION ANALYSIS SHOWED THAT LOWER DNA METHYLATION LEVELS WERE ASSOCIATED WITH PRESENCE OF COPD (CRUDE OR = 0.06, 95% CI 0.00 TO 0.67, P = 0.023). THIS RELATIONSHIP REMAINED SIGNIFICANT AFTER ADJUSTING FOR SEVERAL CONFOUNDERS (OR 0.03, 95% CI 0.00 TO 0.67; P = 0.028). RECEIVER OPERATING CHARACTERISTICS (ROC) CURVE ANALYSIS DEMONSTRATED THE AREA UNDER THE CURVE OF MCYT WAS 0.646, WITH 46.6% SENSITIVITY AND 79.1% SPECIFICITY FOR PRESENCE OF COPD. CONCLUSIONS: THERE WERE NO SIGNIFICANT CORRELATIONS BETWEEN METHYLATION AND OS INDICES. THE PRESENCE AND SEVERITY OF COPD IS ASSOCIATED WITH PROGRESSIVELY LOWER DNA METHYLATION IN BLOOD. HOWEVER, THIS EPIGENETIC ALTERATION SEEMS INDEPENDENT OF OXIDATIVE STRESS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
8  3448  34 HYPERMETHYLATION OF THE N-MYC DOWNSTREAM-REGULATED GENE 2 PROMOTER IN PERIPHERAL BLOOD MONONUCLEAR CELLS IS ASSOCIATED WITH LIVER FIBROSIS IN CHRONIC HEPATITIS B. DNA METHYLATION IS A FUNDAMENTAL EPIGENETIC MODIFICATION TO REGULATE GENE EXPRESSION. N-MYC DOWNSTREAM-REGULATED GENE (NDRG) 2 IS A CYTOPLASMIC PROTEIN AND PARTICIPATES IN THE PATHOGENESIS OF LIVER FIBROSIS. IN THIS STUDY, THE MRNA EXPRESSION AND METHYLATION STATUS OF NDRG2 WAS EVALUATED IN PATIENTS WITH CHRONIC HEPATITIS B (CHB). THE STUDY INCLUDED 143 CHB PATIENTS AND 65 NORMAL CONTROLS (NC). THE MRNA EXPRESSION OF NDRG2 IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) WAS DETECTED BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION. THE METHYLATION STATUS OF THE NDRG2 PROMOTER IN PBMCS WAS DETECTED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. THE NDRG2 MRNA LEVEL WAS LOWER IN THE CHB GROUP THAN IN THE NC GROUP (P < 0.001). METHYLATION FREQUENCY OF THE NDRG2 PROMOTER WAS SIGNIFICANTLY HIGHER IN CHB PATIENTS THAN IN THE NC GROUP (52.44% VS. 26.15%, P < 0.001). IMPORTANTLY, THE RELATIVE EXPRESSION LEVELS OF NDRG2 MRNA WERE SIGNIFICANTLY LOWER IN THE METHYLATED GROUP THAN IN THE UNMETHYLATED GROUP IN BOTH CHB PATIENTS AND NC (P < 0.001). FURTHERMORE, A LOWER MRNA LEVEL AND HYPERMETHYLATION OF NDRG2 WERE ASSOCIATED WITH LIVER FIBROSIS AND INFLAMMATION GRADE IN CHB. THE ASPARTATE AMINOTRANSFERASE-TO-PLATELET RATIO INDEX (APRI) SCORE IS WIDELY USED TO PREDICT LIVER FIBROSIS. THE MRNA EXPRESSION LEVELS AND METHYLATION STATUS OF NDRG2 SHOWED A BETTER SCORE COMPARED TO APRI FOR DISCRIMINATING THE SEVERITY OF LIVER FIBROSIS. IN CONCLUSION, HYPERMETHYLATION OF NDRG2 IN PBMCS WAS CORRELATED WITH DECREASED MRNA EXPRESSION AND WITH LIVER FIBROSIS. THE METHYLATION STATUS OF THE NDRG2 PROMOTER IN PBMCS IS A POTENTIAL NONINVASIVE BIOMARKER TO PREDICT THE SEVERITY OF LIVER FIBROSIS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
9  1953  34 EPIGENETIC AGE ACCELERATION AND CHRONIC HEALTH CONDITIONS AMONG ADULT SURVIVORS OF CHILDHOOD CANCER. BACKGROUND: MOUNTING EVIDENCE SUPPORTS THE OCCURRENCE OF ACCELERATING AGING AMONG LONG-TERM SURVIVORS OF CHILDHOOD CANCER. WE AIMED TO INVESTIGATE EPIGENETIC AGE ACCELERATION (EAA) IN SURVIVORS AND EVALUATE ASSOCIATIONS BETWEEN EAA, TREATMENT EXPOSURES, HEALTH BEHAVIORS, AND CHRONIC HEALTH CONDITIONS (CHCS). METHODS: GENOME-WIDE METHYLATION DATA WERE GENERATED WITH INFINIUM EPIC BEADCHIP ON BLOOD-DERIVED DNA FROM 2139 SURVIVORS AND 282 FREQUENCY MATCHED CONTROLS FROM THE ST JUDE LIFETIME COHORT STUDY. EAAS WERE ESTIMATED AS RESIDUALS FROM A LINEAR REGRESSION OF EPIGENETIC AGE (LEVINE'S CLOCK) AGAINST CHRONOLOGICAL AGE. ADJUSTED LEAST SQUARE MEAN (ALSM) OF EAA WAS CALCULATED AND COMPARED BETWEEN SURVIVORS AND CONTROLS, ACROSS TREATMENT EXPOSURES AND HEALTH BEHAVIORS. ASSOCIATIONS OF EAA WITH 20 CLINICALLY ASSESSED CHCS WERE EVALUATED WITH MULTIVARIABLE PIECEWISE-EXPONENTIAL MODELS. ALL STATISTICAL TESTS FOR P VALUES BELOW WERE 2-SIDED. RESULTS: EAA WAS STATISTICALLY SIGNIFICANTLY HIGHER IN SURVIVORS THAN CONTROLS (ALSM = 0.63, 95% CONFIDENCE INTERVAL [CI] = 0.26 TO 1.01 VS -3.61, 95% CI = -4.43 TO 2.80). IN A MULTIVARIABLE MODEL AMONG SURVIVORS, STATISTICALLY SIGNIFICANTLY HIGHER EAA (P < .05) WAS OBSERVED IN THOSE EXPOSED TO CHEST RADIOTHERAPY, ABDOMEN OR PELVIC RADIOTHERAPY, ALKYLATING AGENTS, GLUCOCORTICOIDS, OR EPIPODOPHYLLOTOXINS. COMPARED WITH SURVIVORS WITH FAVORABLE HEALTH BEHAVIORS (ALSM = 0.26, 95% CI=-0.36 TO 0.87), EAA WAS STATISTICALLY SIGNIFICANTLY HIGHER AMONG SURVIVORS WITH INTERMEDIATE (ALSM = 1.07, 95% CI = 0.59 TO 1.54) OR UNFAVORABLE HEALTH BEHAVIORS (ALSM = 1.45, 95% CI = 0.60 TO 2.30). IN TIME-TO-EVENT ANALYSES, STATISTICALLY SIGNIFICANT ASSOCIATIONS WERE IDENTIFIED BETWEEN EAA TERTILES AND INCIDENCE OF 7 CHCS: HYPERTENSION (3RD VS 1ST TERTILE, RELATIVE RATE [RR] = 1.83, 95% CI = 1.17 TO 2.83), MYOCARDIAL INFARCTION (RR = 2.91, 95% CI = 1.27 TO 7.21), OBESITY (RR = 1.39, 95% CI = 1.17 TO 1.66), OBSTRUCTIVE PULMONARY DEFICIT (RR = 1.86, 95% CI = 0.95 TO 3.77), PERIPHERAL MOTOR NEUROPATHY (RR = 2.89, 95% CI = 1.24 TO 6.97), PERIPHERAL SENSORY NEUROPATHY (RR = 2.04, 95% CI = 0.99 TO 4.26), AND PULMONARY DIFFUSION DEFICITS (RR = 2.75, 95% CI = 0.95 TO 7.63). CONCLUSIONS: EAA IS STATISTICALLY SIGNIFICANTLY HIGHER IN SURVIVORS OF CHILDHOOD CANCER THAN IN NONCANCER CONTROLS AND IS ASSOCIATED WITH SPECIFIC TREATMENT EXPOSURES, UNFAVORABLE HEALTH BEHAVIORS, AND PRESENCE OF SPECIFIC CHCS.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                  
10   401  35 ANALYSIS OF ABERRANT METHYLATION ON PROMOTER SEQUENCES OF TUMOR SUPPRESSOR GENES AND TOTAL DNA IN SPUTUM SAMPLES: A PROMISING TOOL FOR EARLY DETECTION OF COPD AND LUNG CANCER IN SMOKERS. BACKGROUND: CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) IS A DISORDER ASSOCIATED TO CIGARETTE SMOKE AND LUNG CANCER (LC). SINCE EPIGENETIC CHANGES IN ONCOGENES AND TUMOR SUPPRESSOR GENES (TSGS) ARE CLEARLY IMPORTANT IN THE DEVELOPMENT OF LC. IN THIS STUDY, WE HYPOTHESIZE THAT TOBACCO SMOKERS ARE SUSCEPTIBLE FOR METHYLATION IN THE PROMOTER REGION OF TSGS IN AIRWAY EPITHELIAL CELLS WHEN COMPARED WITH NON-SMOKER SUBJECTS. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE THE USEFULNESS OF DETECTION OF GENES PROMOTER METHYLATION IN SPUTUM SPECIMENS, AS A COMPLEMENTARY TOOL TO IDENTIFY LC BIOMARKERS AMONG SMOKERS WITH EARLY COPD. METHODS: WE DETERMINED THE AMOUNT OF DNA IN INDUCED SPUTUM FROM PATIENTS WITH COPD (N = 23), LC (N = 26), AS WELL AS IN HEALTHY SUBJECTS (CTR) (N = 33), USING A COMMERCIAL KIT FOR DNA PURIFICATION, FOLLOWED BY ABSORBANCE MEASUREMENT AT 260 NM. THE FREQUENCY OF CDKN2A, CDH1 AND MGMT PROMOTER METHYLATION IN THE SAME GROUPS WAS DETERMINED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP). THE FISHER'S EXACT TEST WAS EMPLOYED TO COMPARE FREQUENCY OF RESULTS BETWEEN DIFFERENT GROUPS. RESULTS: DNA CONCENTRATION WAS 7.4 AND 5.8 TIMES HIGHER IN LC AND COPD COMPARED TO THE (CTR) (P < 0.0001), RESPECTIVELY. METHYLATION STATUS OF CDKN2A AND MGMT WAS SIGNIFICANTLY HIGHER IN COPD AND LC PATIENTS COMPARED WITH CTR GROUP (P < 0.0001). FREQUENCY OF CDH1 METHYLATION ONLY SHOWED A STATISTICALLY SIGNIFICANT DIFFERENCE BETWEEN LC PATIENTS AND CTR GROUP (P < 0.05). CONCLUSIONS: WE PROVIDE EVIDENCE THAT ABERRANT METHYLATION OF TSGS IN SAMPLES OF INDUCED SPUTUM IS A USEFUL TOOL FOR EARLY DIAGNOSTIC OF LUNG DISEASES (LC AND COPD) IN SMOKER SUBJECTS. VIRTUAL SLIDES: THE ABSTRACT MUST FINISH WITH THE FOLLOWING TEXT: VIRTUAL SLIDES THE VIRTUAL SLIDE(S) FOR THIS ARTICLE CAN BE FOUND HERE: HTTP://WWW.DIAGNOSTICPATHOLOGY.DIAGNOMX.EU/VS/1127865005664160.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
11  4246  33 METHYLATION STATUS OF THE T-CADHERIN GENE PROMOTOR IN PERIPHERAL BLOOD MONONUCLEAR CELLS IS ASSOCIATED WITH HBV-RELATED HEPATOCELLULAR CARCINOMA PROGRESSION. DNA METHYLATION IS ONE OF THE EPIGENETIC MECHANISMS TO REGULATE GENE EXPRESSION AND FREQUENTLY OCCURS IN HUMAN CANCER CELLS. T-CADHERIN (CDH13) IS A NEW MEMBER OF THE CADHERIN SUPERFAMILY AND POSSESSES MULTIPLE FUNCTIONS. OUR STUDY INCLUDED 26 NORMAL CONTROLS (NCS), 65 CHRONIC HEPATITIS B PATIENTS (CHB), 14 LIVER CIRRHOSIS PATIENTS (LC) AND 157 HEPATOCELLULAR CARCINOMA PATIENTS (HCC). WE MAINLY FOCUSED ON THE MRNA EXPRESSION AND METHYLATION STATUS OF CDH13 IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS), WHICH WERE DETECTED BY SEMI-QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (RT-QPCR) AND METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP) RESPECTIVELY. THE CDH13 MRNA LEVEL WAS LOWER IN HCC, ESPECIALLY IN EARLY-STAGE OF HCC THAN IN NCS AND CHB GROUPS (P < 0.05). METHYLATION FREQUENCY OF THE CDH13 PROMOTER WAS SIGNIFICANTLY HIGHER IN HCC PATIENTS THAN IN THE NCS AND CHB GROUPS (67.52 % VS 0.00 %, P < 0.001, 67.52 % VS 52.31 %, P < 0.05, RESPECTIVELY). CDH13 MRNA LEVEL WAS SIGNIFICANTLY AND RELATIVELY LOWER IN METHYLATED GROUPS THAN IN UNMETHYLATED GROUPS AMONG THE WHOLE PARTICIPANTS. THE METHYLATION LEVEL OF CDH13 PROMOTER IN HCC MIGHT BE INFLUENCED OR PARTLY INFLUENCED BY SOME CRITICAL FACTORS SUCH AS TBIL, ALB AND AFP (P < 0.05). AS AN IMPORTANT FACTOR IN SIGNALING PATHWAY REGULATING BY CDH13 TO PROMOTE CARCINOGENESIS, JNK LEVEL WAS SIGNIFICANTLY HIGHER IN HCC WHICH HAD A HIGHER METHYLATION FREQUENCY THAN IN NCS, CHB AND LC (P < 0.05). FURTHERMORE, THE COMBINATION OF THE METHYLATED CDH13 LEVEL AND AFP LEVEL SHOWED A BETTER SCORE: AUC = 0.796 (SE = 0.031, 95 %CI 0.735-0.857; P < 0.001) IN MALE AND AUC = 0.832 (SE = 0.057, 95 %CI 0.721-0.944; P < 0.001) IN FEMALE COMPARED TO AFP ALONE FOR DIAGNOSING HCC FROM NCS, CHB AND LC. THE METHYLATION OF CDH13 PROMOTER WAS AN INDEPENDENT PREDICTOR FOR ASSESSING THE PROGNOSIS OF HCC PATIENTS (R=-1.378 P < 0.05). IN CONCLUSION, HYPERMETHYLATION OF CDH13 IN PBMCS WAS ASSOCIATED WITH THE UNDEREXPRESSION OF MRNA AND THE HIGH RISK OF HCC. THE METHYLATION STATUS OF THE CDH13 PROMOTER IN PBMCS WAS A POTENTIAL NONINVASIVE BIOMARKER TO PREDICT THE PROGNOSIS OF HCC PATIENTS.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
12  6415  30 THE STUDY OF P16 AND P15 GENE METHYLATION IN HEAD AND NECK SQUAMOUS CELL CARCINOMA AND THEIR QUANTITATIVE EVALUATION IN PLASMA BY REAL-TIME PCR. EPIGENETIC SILENCING OF THE P16 AND P15 GENES BY PROMOTER METHYLATION ARE COMMONLY OBSERVED IN HUMAN EPITHELIAL MALIGNANCIES, INCLUDING HEAD AND NECK SQUAMOUS CELL CARCINOMAS (HNSCC). IN THIS STUDY, A METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP) WAS USED TO EVALUATE THE METHYLATION STATUS OF THE P16 AND P15 GENES IN 73 HNSCC SURGICAL SPECIMENS. P16 AND P15 GENE METHYLATION WAS ALSO EXAMINED IN 29 PAIRED METASTATIC LYMPH NODES AND 29 PAIRED HISTOLOGICALLY, NORMAL RESECTION MARGIN MUCOSAE. THE QUANTITY OF CELL-FREE METHYLATED P16 AND P15 DNA IN THE PLASMA SAMPLES OF 20 HNSCC PATIENTS AND 24 HEALTHY CONTROLS WAS ALSO EXAMINED USING A FLUORESCENCE-BASED REAL-TIME PCR ASSAY. THE FREQUENCIES OF P16 AND P15 METHYLATION IN THE PRIMARY TUMOUR WERE 49% AND 60%, RESPECTIVELY. CONCORDANT METHYLATION OF P16 AND P15 IN TUMOUR SAMPLES AND METASTATIC LYMPH NODES WAS FOUND IN 59 AND 38% OF CASES, RESPECTIVELY. A SIGNIFICANTLY HIGHER PREVALENCE OF P15 METHYLATION WAS FOUND IN HISTOLOGICALLY-NORMAL SURGICAL MARGIN EPITHELIA OF HNSCC PATIENTS WITH CHRONIC SMOKING AND DRINKING HABITS COMPARED WITH NON-SMOKERS AND NON-DRINKERS. IN ADDITION, METHYLATED P16 AND P15 DNA LEVELS WERE SIGNIFICANTLY HIGHER IN THE PLASMA OF HNSCC PATIENTS (MEAN 56 COPIES/ML PLASMA AND 65 COPIES/ML PLASMA, RESPECTIVELY) COMPARED WITH NORMAL CONTROLS (MEAN 6 COPIES/ML PLASMA AND 16 COPIES/ML PLASMA, RESPECTIVELY). IN CONCLUSION, PROMOTER METHYLATION OF THE P16 AND P15 GENES IS INVOLVED IN THE PATHOGENESIS OF HNSCC AND MAY BE RELATED TO CHRONIC SMOKING AND DRINKING. THE DIFFERENTIAL LEVELS OF METHYLATED P16 AND P15 DNA IN PLASMA MIGHT BE POTENTIAL USEFUL MARKERS IN SCREENING HIGH-RISK POPULATIONS FOR EARLY HNSCC AND MONITORING THEIR TREATMENT RESPONSE.	2003	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
13  2326  33 EPIGENETIC REGULATION OF HOTAIR IN ADVANCED CHRONIC MYELOID LEUKEMIA. PURPOSE: CHRONIC MYELOID LEUKEMIA (CML) ACCOUNTS FOR ~10% OF LEUKEMIA CASES, AND ITS PROGRESSION INVOLVES EPIGENETIC GENE REGULATION. THIS STUDY INVESTIGATED EPIGENETIC REGULATION OF HOTAIR AND ITS TARGET MICRORNA, MIR-143, IN ADVANCED CML. PATIENTS AND METHODS: WE FIRST ISOLATED BONE MARROW MONONUCLEAR CELLS FROM 70 PATIENTS WITH DIFFERENT PHASES OF CML AND FROM HEALTHY DONORS AS NORMAL CONTROL; WE ALSO CULTURED K562 AND KCL22 CELLS, TREATED WITH DEMETHYLATION DRUG; MTT ASSAY, FLOW CYTOMETRY, QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QPCR), METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP), WESTERN BLOT, LUCIFERASE ASSAY, RNA PULL-DOWN ASSAY AND RNA-BINDING PROTEIN IMMUNOPRECIPITATION (RIP) ASSAY WERE PERFORMED. RESULT: AS MEASURED BY QPCR, HOTAIR EXPRESSION IN K562 CELLS, KCL22 CELLS, AND SAMPLES FROM CASES OF ADVANCED-STAGE CML INCREASED WITH LEVELS OF SEVERAL DNA METHYLTRANSFERASES AND HISTONE DEACETYLATES, INCLUDING DNMT1, DNMT3A, HDAC1, EZH2, AND LSD1, AND MIR-143 LEVELS WERE DECREASED AND HOTAIR LEVELS WERE INCREASED. TREATMENT WITH 5-AZACYTIDINE, A DNA METHYLATION INHIBITOR, DECREASED DNMT1, DNMT3A, HDAC1, EZH2, LSD1 MRNA, PROTEIN LEVELS, AND HOTAIR MRNA LEVELS BUT INCREASED MIR-143 LEVELS. HOTAIR KNOCKDOWN AND MIR-143 OVEREXPRESSION BOTH INHIBITED PROLIFERATION AND PROMOTED APOPTOSIS IN KCL22 AND K562 CELLS THROUGH THE PI3K/AKT PATHWAY. RNA PULL-DOWN, MASS SPECTROMETRY, AND RIP ASSAYS SHOWED THAT HOTAIR INTERACTED WITH EZH2 AND LSD1. A DUAL-LUCIFERASE ASSAY DEMONSTRATED THAT HOTAIR INTERACTED WITH MIR-143. CONCLUSION: OUR FINDINGS DEMONSTRATE THE KEY EPIGENETIC INTERACTIONS OF HOTAIR RELATED TO CML PROGRESSION AND SUGGEST HOTAIR AS A POTENTIAL THERAPEUTIC TARGET FOR ADVANCED CML. FURTHERMORE, OUR RESULTS SUPPORT THE USE OF DEMETHYLATION DRUGS AS A CML TREATMENT STRATEGY.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
14  1065  49 CLINICAL SIGNIFICANCE OF SERUM DRAM1 MRNA, ARSA MRNA, HSA-MIR-2053 AND LNCRNA-RP1-86D1.3 AXIS EXPRESSION IN MALIGNANT PLEURAL MESOTHELIOMA. AIM AND BACKGROUND: MALIGNANT PLEURAL MESOTHELIOMA (MPM) IS A LETHAL CANCER MAINLY CAUSED BY CHRONIC EXPOSURE OF ASBESTOS. IN THIS PILOT STUDY, WE AIMED TO ASSESS THE EXPRESSION OF SERUM RNA-BASED BIOMARKER PANEL EXPLORING THEIR CLINICAL UTILITY AS DIAGNOSTIC AND PROGNOSTIC BIOMARKERS FOR MPM. METHODS: WE HAVE SELECTED AN MPM-SPECIFIC RNA-BASED BIOMARKER PANEL THROUGH BIOINFORMATICS ANALYSIS BASED ON THE INTEGRATION OF DNA DAMAGE REGULATED AUTOPHAGY MODULATOR 1 (DRAM1) AND ARYLSULFATASE A ( ARSA) GENE EXPRESSION WITH THEIR EPIGENETIC REGULATORS MICRORNA ( MIR-2053) AND LONG NONCODING RNA ( LNCRNA-RP1-86D1.3). THEN, QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QPCR) VALIDATION IN SERA OF 60 MPM PATIENTS, 20 CHRONIC ASBESTOS EXPOSURE PATIENTS, AND 20 HEALTHY VOLUNTEERS WAS DONE. LASTLY, THE PROGNOSTIC POWER OF THE SELECTED PANEL WAS ASSESSED. RESULTS: THE EXPRESSION OF SERUM DRAM1 MESSENGER RNA (MRNA), ARSA MRNA, HSA-MIR-2053 AND LNCRNA-RP1-86D1.3 WERE POSITIVE IN 78.3%, 90%, 85%, AND 83.3% OF MPM PATIENTS, RESPECTIVELY. THE RNA-BASED BIOMARKER PANEL WAS ABLE TO DISCRIMINATE BETWEEN MPM PATIENTS AND CONTROLS WITH HIGH ACCURACY AND THEIR COMBINED SENSITIVITY REACHED 100% FOR THE DIAGNOSIS OF MPM. KAPLAN-MEIER ANALYSIS SHOWED THAT HSA-MIR-2053 IS AN INDEPENDENT PROGNOSTIC FACTOR OF MPM. CONCLUSION: OUR PRELIMINARY DATA REVEALED THAT THE CHOSEN RNAS PLAY AN IMPORTANT ROLE IN DRIVING MPM DEVELOPMENT AND PROGRESSION.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
15   576  25 BDNF METHYLATION AND SUICIDAL IDEATION IN PATIENTS WITH ACUTE CORONARY SYNDROME. OBJECTIVE: PATIENTS WITH ACUTE CORONARY SYNDROME (ACS) ARE AT AN INCREASED RISK OF SUICIDE. IT IS WELL KNOWN THAT EPIGENETIC MECHANISMS MAY EXPLAIN THE PATHOPHYSIOLOGY OF SUICIDAL BEHAVIOR INCLUDING SUICIDAL IDEATION (SI), BUT NO STUDY HAS EXPLORED THESE MECHANISMS IN ACS POPULATIONS. METHODS: IN TOTAL, 969 PATIENTS WERE INITIALLY RECRUITED WITHIN 2 WEEKS OF THE ACUTE CORONARY EVENT AND, 711 PATIENTS WERE SUCCESSFULLY FOLLOWED UP 1 YEAR AFTER ACS. SI WAS EVALUATED USING THE RELEVANT ITEMS ON THE MONTGOMERY-ASBERG DEPRESSION RATING SCALE AND COVARIATES POTENTIALLY AFFECTING SI WERE ESTIMATED. RESULTS: BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) HYPERMETHYLATION WAS ASSOCIATED WITH SI IN BOTH THE ACUTE AND CHRONIC PHASES OF ACS, ALTHOUGH THE ASSOCIATION WAS NOT STATISTICALLY SIGNIFICANT IN THE ACUTE PHASE AFTER APPLYING BONFERRONI'S CORRECTION. CONCLUSION: THESE RESULTS SUGGESTED THAT BDNF HYPERMETHYLATION MAY HAVE PLAYED A ROLE IN AN EPIGENETIC PREDISPOSITION FOR SI IN ACS PATIENTS, PARTICULARLY DURING THE CHRONIC PHASE.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
16  1029  35 CIRCULATING PLASMA MICRORNA IN PATIENTS WITH ACTIVE ACROMEGALY. CONTEXT: EXCESSIVE PRODUCTION OF GROWTH HORMONE CAUSES MARKED MULTIORGAN CHANGES IN PATIENTS WITH ACROMEGALY, WHICH MAY INVOLVE EPIGENETIC MECHANISMS. OBJECTIVE: TO EVALUATE DIFFERENCES IN CIRCULATING MICRORNAS (MIRNAS) ASSOCIATED WITH CHRONIC GROWTH HORMONE OVERPRODUCTION IN ADULTS. DESIGN AND SETTING: A CROSS-SECTIONAL CASE-CONTROL STUDY WAS CONDUCTED AT A TERTIARY MEDICAL CENTER. PARTICIPANTS: WE ENROLLED 12 CONSECUTIVE PATIENTS WITH ACROMEGALY ALONG WITH 12 AGE- AND SEX-MATCHED CONTROLS IN THE DISCOVERY PHASE OF THE STUDY AND THEN EXTENDED THIS COHORT TO 47 PATIENTS WITH ACROMEGALY AND 28 HEALTHY CONTROLS FOR THE VALIDATION STUDY. MAIN OUTCOME MEASURES: PLASMA MIRNAS WERE QUANTIFIED BY NEXT-GENERATION SEQUENCING (NGS) IN THE DISCOVERY PHASE. LEVELS OF SELECTED MIRNAS WERE VALIDATED ON EXTENDED COHORTS USING REVERSE TRANSCRIPTION QUANTITATIVE POLYMERASE CHAIN REACTION (RT-QPCR), COMPARED BETWEEN GROUPS, AND CORRELATED WITH CLINICAL PARAMETERS. RESULTS: BASED ON NGS DATA, WE SELECTED 3 PLASMA MIRNAS DOWNREGULATED IN PATIENTS WITH ACROMEGALY COMPARED TO HEALTHY CONTROLS: MIR-4446-3P -1.317 (P = 0.001), MIR-215-5P -3.040 (P = 0.005), AND MIR-342-5P -1.875 (P = 0.013) WITHOUT MULTIPLICITY CORRECTION FOR ALL 3 MIRNAS. THESE RESULTS WERE CONFIRMED BY RT-QPCR IN THE VALIDATION PHASE FOR 2 MIRNAS OUT OF 3: MIR-4446-3P (P < 0.001, PADJUSTED < 0.001), AREA UNDER THE RECEIVER-OPERATOR CURVE (AUC) 0.862 (95% CI 0.723-0.936; P < 0.001) AND MIR-215-5P (P < 0.001, PADJUSTED < 0.001), AUC 0.829 (95% CI 0.698-0.907; P < 0.001) TO DIFFERENTIATE PATIENTS WITH ACROMEGALY FROM HEALTHY CONTROLS. CONCLUSIONS: IN A 2-PHASE EXPERIMENT USING 2 DIFFERENT TECHNIQUES WE FOUND AND VALIDATED THE DOWNREGULATION OF PLASMA MIR-4446-3P AND MIR-215-5P IN PATIENTS WITH ACROMEGALY COMPARED TO HEALTHY SUBJECTS, WHICH MAKES THEM PROMISING BIOMARKERS FOR FURTHER RESEARCH.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
17  1074  29 CLONAL HAEMATOPOIESIS IN CHRONIC ISCHAEMIC HEART FAILURE: PROGNOSTIC ROLE OF CLONE SIZE FOR DNMT3A- AND TET2-DRIVER GENE MUTATIONS. AIMS: SOMATIC MUTATIONS OF THE EPIGENETIC REGULATORS DNMT3A AND TET2 CAUSING CLONAL EXPANSION OF HAEMATOPOIETIC CELLS (CLONAL HAEMATOPOIESIS; CH) WERE SHOWN TO BE ASSOCIATED WITH POOR PROGNOSIS IN CHRONIC ISCHAEMIC HEART FAILURE (CHF). THE AIM OF OUR ANALYSIS WAS TO DEFINE A THRESHOLD OF VARIANT ALLELE FREQUENCY (VAF) FOR THE PROGNOSTIC SIGNIFICANCE OF CH IN CHF. METHODS AND RESULTS: WE ANALYSED BONE MARROW AND PERIPHERAL BLOOD-DERIVED CELLS FROM 419 PATIENTS WITH CHF BY ERROR-CORRECTED AMPLICON SEQUENCING. CUT-OFF VAFS WERE OPTIMIZED BY MAXIMIZING SENSITIVITY PLUS SPECIFICITY FROM A TIME-DEPENDENT RECEIVER OPERATING CHARACTERISTIC (ROC) CURVE ANALYSIS FROM CENSORED DATA. 56.2% OF PATIENTS WERE CARRIERS OF A DNMT3A- (N = 173) OR A TET2- (N = 113) MUTATION WITH A VAF >0.5%, WITH 59 PATIENTS HARBOURING MUTATIONS IN BOTH GENES. SURVIVAL ROC ANALYSES REVEALED AN OPTIMIZED CUT-OFF VALUE OF 0.73% FOR TET2- AND 1.15% FOR DNMT3A-CH-DRIVER MUTATIONS. FIVE-YEAR-MORTALITY WAS 18% IN PATIENTS WITHOUT ANY DETECTED DNMT3A- OR TET2 MUTATION (VAF < 0.5%), 29% WITH ONLY ONE DNMT3A- OR TET2-CH-DRIVER MUTATIONS ABOVE THE RESPECTIVE CUT-OFF LEVEL AND 42% IN PATIENTS HARBOURING BOTH DNMT3A- AND TET2-CH-DRIVER MUTATIONS ABOVE THE RESPECTIVE CUT-OFF LEVELS. IN CARRIERS OF A DNMT3A MUTATION WITH VAF >/= 1.15%, 5-YEAR MORTALITY WAS 31%, COMPARED WITH 18% MORTALITY IN THOSE WITH VAF < 1.15% (P = 0.048). LIKEWISE, IN PATIENTS WITH TET2 MUTATIONS, 5-YEAR MORTALITY WAS 32% WITH VAF >/= 0.73%, COMPARED WITH 19% MORTALITY WITH VAF < 0.73% (P = 0.029). CONCLUSION: THE PRESENT STUDY DEFINES NOVEL THRESHOLD LEVELS FOR CLONE SIZE CAUSED BY ACQUIRED SOMATIC MUTATIONS IN THE CH-DRIVER GENES DNMT3A AND TET2 THAT ARE ASSOCIATED WITH WORSE OUTCOME IN PATIENTS WITH CHF.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
18  3294  26 HIGH INCIDENCE OF MGMT AND RARBETA PROMOTER METHYLATION IN PRIMARY GLIOBLASTOMAS: ASSOCIATION WITH HISTOPATHOLOGICAL CHARACTERISTICS, INFLAMMATORY MEDIATORS AND CLINICAL OUTCOME. GLIOBLASTOMAS, THE MOST FREQUENT PRIMARY BRAIN TUMORS IN ADULTS, ARE CHARACTERIZED BY A HIGHLY AGGRESSIVE, INFLAMMATORY AND ANGIOGENIC PHENOTYPE. METHYLATION OF CPG ISLANDS IN CANCER-RELATED GENES MAY SERVE AS AN EPIGENETIC BIOMARKER FOR GLIOBLASTOMA DIAGNOSIS AND PROGNOSIS. THE AIM OF THIS STUDY WAS TO ANALYZE THE METHYLATION STATUS OF FOUR CRITICAL TUMOR-ASSOCIATED GENES (MGMT, RARBETA, RASSF1A, CDH13), AND INVESTIGATE POSSIBLE LINKS WITH INFLAMMATORY (INTERLEUKIN [IL]-6, IL-8) AND ANGIOGENIC MEDIATORS (VASCULAR ENDOTHELIAL GROWTH FACTOR [VEGF], CYCLOOXYGENASE [COX]-2) AND CLINICAL OUTCOME IN 23 GLIOMA SAMPLES (6 GRADE II ASTROCYTOMAS, 17 GRADE IV GLIOBLASTOMAS). RARBETA AND MGMT GENES WERE MORE FREQUENTLY METHYLATED IN 70.58% AND 58.8% OF GLIOBLASTOMAS, RESPECTIVELY. RASSF1A AND CDH13 DISPLAYED A SIMILAR METHYLATION FREQUENCY (23.52%) IN GLIOBLASTOMAS. NO GENE METHYLATION WAS OBSERVED IN GRADE II ASTROCYTOMAS. TUMOR GRADE CORRELATED POSITIVELY WITH MGMT AND RARBETA METHYLATION (P = 0.005 AND P = 0.019, RESPECTIVELY) AND THE EXTENT OF NECROSIS (P = 0.001 AND P = 0.003). INTERESTINGLY, THE MARKER OF CHRONIC INFLAMMATION, IL-6, WAS POSITIVELY ASSOCIATED WITH METHYLATION OF MGMT (P = 0.004), RARBETA (P = 0.002), AND RASSF1A (P = 0.0081) AS WELL AS THE TOTAL NUMBER OF METHYLATED GENES (P < 0.0001), INDICATING THE IMPORTANT ROLE OF IL-6 IN MAINTAINING PROMOTER METHYLATION OF THESE GENES. VEGF EXPRESSION CORRELATED POSITIVELY WITH MGMT AND RARBETA METHYLATION ALTHOUGH THESE RELATIONSHIPS WERE OF MARGINAL SIGNIFICANCE (P = 0.0679 AND P = 0.0757). KAPLAN-MEIER UNIVARIATE SURVIVAL ANALYSIS INDICATED AN UNFAVORABLE SURVIVAL PERIOD IN PATIENTS WITH MGMT METHYLATION COMPARED WITH THOSE WITHOUT METHYLATION (P = 0.0474). OUR STUDY HIGHLIGHTS THE IMPLICATION OF MGMT AND RARBETA METHYLATION IN THE AGGRESSIVE PHENOTYPE OF PRIMARY GLIOBLASTOMAS. THE ASSOCIATION OF MGMT METHYLATION WITH CLINICAL OUTCOME INDICATES ITS POTENTIAL PROGNOSTIC VALUE.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
19  1064  38 CLINICAL SIGNIFICANCE OF PROMOTER METHYLATION STATUS OF TUMOR SUPPRESSOR GENES IN CIRCULATING DNA OF PANCREATIC CANCER PATIENTS. INTRODUCTION: PANCREATIC DUCTAL ADENOCARCINOMA (PDAC) IS A VERY AGGRESSIVE CANCER. THERE ARE VARIOUS SUB-CELLULAR EVENTS (BOTH GENETIC AND EPIGENETIC) THAT GET DYSREGULATED LEADING TO TUMORIGENESIS. METHYLATION IN PROMOTERS OF TUMOR SUPPRESSOR GENES IS ONE OF THESE EPIGENETIC PHENOMENA CONTRIBUTING TO THE PATHOGENESIS OF CANCER. GENES ANALYZED FOR PROMOTER METHYLATION STATUS IN THIS STUDY NAMELY SPARC (SECRETED PROTEIN ACIDIC AND RICH IN CYSTEINE, UCHL1 (UBIQUITIN CARBOXY-TERMINAL HYDROLASE L1), NPTX2 (NEURONAL PENTRAXIN 2), PENK (PROENKEPHALIN) HAD BEEN STUDIED IN PANCREATIC CANCER, BUT THERE IS A NEED TO CHECK METHYLATION IN THESE GENES AS CIRCULATORY NON-INVASIVE MARKERS. THIS STUDY ANALYZED THE ABSOLUTE QUANTIFICATION OF METHYLATION LEVELS OF SPARC, UCHL1, PENK, AND NPTX2 GENES PROMOTERS IN PDAC PATIENTS AS WELL AS IN CHRONIC PANCREATITIS (CP) PATIENTS AND HEALTHY SUBJECTS (HC) AND EVALUATED ITS CLINICAL SIGNIFICANCE IN PDAC. MATERIALS AND METHODS: THE STUDY INCLUDED 65 PDAC PATIENTS, 25 CP PATIENTS, AND 25 HEALTHY CONTROLS. DNA WAS EXTRACTED FROM THEIR PLASMA SAMPLES AND SUBSEQUENTLY GIVEN BISULFITE TREATMENT. ABSOLUTE QUANTIZATION OF METHYLATED AND UNMETHYLATED COPIES OF GENE PROMOTERS OF ALL THE FOUR GENES WAS PERFORMED USING REAL-TIME PCR (SYBR GREEN) BY THE STANDARD CURVE METHOD. METHYLATION LEVELS WERE EXPRESSED AS METHYLATION INDEX (MI) FOR EACH GENE IN EACH PATIENT. MI WAS CALCULATED FROM ABSOLUTE COPY NUMBERS AS FOLLOWS: MI-METHYLATED COPY NUMBER/METHYLATED COPY NUMBER + UNMETHYLATED COPY NUMBER). THESE INDICES WERE USED TO COMPARE GENE METHYLATION LEVELS WITHIN DIFFERENT GROUPS AND TO CORRELATE WITH CLINICOPATHOLOGICAL FEATURES AND SURVIVAL OF PANCREATIC CANCER PATIENTS. AN APPROPRIATE STATISTICAL ANALYSIS WAS APPLIED. RESULTS: METHYLATION INDICES FOR ALL THE FOUR GENES IN PDAC CASES WERE FOUND TO BE SIGNIFICANTLY HIGHER AS COMPARED TO THAT IN HEALTHY INDIVIDUALS. SPARC MI VALUES WERE FOUND TO DIFFERENTIATE EARLY-STAGE PDAC PATIENTS FROM CP PATIENTS. PDAC PATIENTS WITH THE METASTASIZED DISEASE AND STAGE IV DISEASE WERE FOUND TO HAVE HIGH MI FOR THE SPARC GENE AS WELL AS FOR THE NPTX2 GENE, WHILE A HIGHER UCHL1 METHYLATION INDEX WAS FOUND TO CORRELATE WITH AN ADVANCED STAGE OF THE DISEASE. HIGHER MI VALUES FOR SPARC AND NPTX2 GENES WERE FOUND TO ASSOCIATE WITH POOR SURVIVAL IN PATIENTS WITH PDAC. CONCLUSION: METHYLATION LOAD IN THE FORM OF MI FOR EACH OF THE FOUR GENES ASSESSED IN PLASMA MAY EMERGE AS A NON-INVASIVE BIOMARKER TO DIFFERENTIATE PANCREATIC CANCER FROM HEALTHY INDIVIDUALS. BUT ONLY SPARC AND NPTX2 HYPERMETHYLATION WERE ABLE TO DISTINGUISH PANCREATIC CANCER FROM CHRONIC PANCREATITIS. ASSOCIATION OF ABERRANT METHYLATION IN SPARC AND NPTX2 GENE WITH METASTASIS AND POOR SURVIVAL OF PATIENTS SUGGEST THE ROLE OF METHYLATION IN THESE GENES AS PROGNOSTIC MARKERS.	2020	

20  4231  30 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