1   447 144 APABETALONE LOWERS SERUM ALKALINE PHOSPHATASE AND IMPROVES CARDIOVASCULAR RISK IN PATIENTS WITH CARDIOVASCULAR DISEASE. BACKGROUND AND AIMS: IN PATIENTS WITH CARDIOVASCULAR DISEASE, CONSIDERABLE RESIDUAL RISK REMAINS DESPITE EVIDENCE-BASED SECONDARY PREVENTION MEASURES. ALKALINE PHOSPHATASE (ALP) HAS BEEN SUGGESTED AS A MODIFIABLE CARDIOVASCULAR RISK FACTOR. WE SOUGHT TO DETERMINE WHETHER CARDIOVASCULAR RISK REDUCTION BY THE BROMODOMAIN AND EXTRA-TERMINAL (BET) PROTEIN INHIBITOR APABETALONE IS ASSOCIATED WITH THE CONCOMITANT LOWERING OF SERUM ALP. METHODS: IN A POST-HOC ANALYSIS OF 795 PATIENTS WITH ESTABLISHED CORONARY HEART DISEASE AND STATIN TREATMENT, WHO PARTICIPATED IN PHASE 2 PLACEBO-CONTROLLED TRIALS OF APABETALONE, WE DETERMINED THE EFFECT OF ASSIGNED TREATMENT FOR UP TO 24 WEEKS ON THE INCIDENCE OF MAJOR ADVERSE CARDIOVASCULAR EVENTS (MACE) AND SERUM ALP. RESULTS: BASELINE ALP (MEDIAN 72 U/L) PREDICTED MACE (DEATH, NON-FATAL MYOCARDIAL INFARCTION, CORONARY REVASCULARIZATION, OR HOSPITALIZATION FOR CARDIOVASCULAR CAUSES), INDEPENDENT OF HIGH-SENSITIVITY C-REACTIVE PROTEIN (HSCRP), SEX, AGE, RACE, STUDY, CARDIOVASCULAR RISK FACTORS, CHRONIC KIDNEY DISEASE (CKD), LIVER FUNCTION MARKERS AND TREATMENT ALLOCATION (HAZARD RATIO [HR] PER STANDARD DEVIATION [SD] 1.6, 95% CI 1.19-2.16, P = 0.002). MEAN PLACEBO-CORRECTED DECREASES IN ALP FROM BASELINE WERE 9.2% (P < 0.001) AFTER 12-14 WEEKS AND 7.7% (P < 0.001) AFTER 24-26 WEEKS OF APABETALONE TREATMENT. IN THE APABETALONE GROUP, A 1-SD REDUCTION IN ALP WAS ASSOCIATED WITH A HR FOR MACE OF 0.64 (95% CI 0.46-0.90, P = 0.009). CONCLUSIONS: SERUM ALP PREDICTS RESIDUAL CARDIOVASCULAR RISK, INDEPENDENT OF HSCRP, ESTABLISHED CARDIOVASCULAR RISK FACTORS AND CKD, IN PATIENTS WITH CARDIOVASCULAR DISEASE ON STATIN TREATMENT. APABETALONE LOWERS SERUM ALP, WHICH WAS ASSOCIATED WITH A LOWER RISK OF CARDIOVASCULAR EVENTS. WHETHER THE BENEFICIAL CARDIOVASCULAR EFFECTS OF APABETALONE ARE CAUSALLY RELATED TO ALP REDUCTION REMAINS UNDETERMINED.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
2   448  52 APABETALONE MEDIATED EPIGENETIC MODULATION IS ASSOCIATED WITH FAVORABLE KIDNEY FUNCTION AND ALKALINE PHOSPHATASE PROFILE IN PATIENTS WITH CHRONIC KIDNEY DISEASE. BACKGROUND/AIMS: THE ASSOCIATION BETWEEN SERUM ALKALINE PHOSPHATASE (ALP) WITH ADVERSE CARDIOVASCULAR OUTCOMES, IN CHRONIC KIDNEY DISEASE (CKD) PATIENTS HAS PREVIOUSLY BEEN REPORTED AND MAY BE A RESULT OF INCREASED VASCULAR CALCIFICATION AND INFLAMMATION. HERE WE REPORT, FOR THE FIRST TIME, THE EFFECTS OF PHARMACOLOGIC EPIGENETIC MODULATION ON LEVELS OF ALP AND KIDNEY FUNCTION VIA A NOVEL ORAL SMALL MOLECULE BET INHIBITOR, APABETALONE, IN CKD PATIENTS. METHODS: A POST-HOC ANALYSIS EVALUATED PATIENTS WITH ESTIMATED GLOMERULAR FILTRATION RATE (EGFR) <60 ML/MIN/1.73M2, WHO PARTICIPATED IN THE APABETALONE PHASE 2 RANDOMIZED CONTROLLED TRIALS (SUSTAIN AND ASSURE). 48 CKD SUBJECTS WITH A HISTORY OF CARDIOVASCULAR DISEASE (CVD) WERE TREATED WITH 100MG TWICE-DAILY OF 24 AND 26 WEEKS OF APABETALONE OR PLACEBO. ALP AND EGFR WERE MEASURED PRIOR TO RANDOMIZATION AND AT FINAL VISITS. RESULTS: PATIENTS WHO RECEIVED APABETALONE (N=35) VERSUS PLACEBO (N=13) OVER 6 MONTHS SHOWED SIGNIFICANTLY (P=0.02) LOWERED SERUM ALP -14.0% (P<0.0001 VERSUS BASELINE) VERSUS -6.3% (P=0.9 VERSUS BASELINE). THE EGFR IN THE APABETALONE GROUP INCREASED BY 3.4% (1.7 ML/MIN/1.73 M2) (P=0.04 VERSUS BASELINE) AND DECREASED BY 5.8% (2.9 ML/MIN/1.73 M2) (P=0.6 VERSUS BASELINE) IN THE PLACEBO GROUP. APABETALONE WAS WELL TOLERATED. CONCLUSION: A POST-HOC ANALYSIS OF CKD SUBJECTS FROM THE SUSTAIN AND ASSURE RANDOMIZED CONTROLLED TRIALS DEMONSTRATED FAVORABLE EFFECTS OF APABETALONE ON ALP AND EGFR, AND GENERATED THE HYPOTHESIS THAT EPIGENETIC MODULATION BY BET INHIBITION MAY POTENTIALLY OFFER A NOVEL THERAPEUTIC STRATEGY TO TREAT CVD AND PROGRESSIVE KIDNEY FUNCTION LOSS IN CKD PATIENTS. THIS IS BEING EXAMINED IN THE PHASE III TRIAL BETONMACE.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
3  6043  47 THE COMBINED PROGNOSTIC SIGNIFICANCE OF ALKALINE PHOSPHATASE AND INTRACRANIAL ARTERIAL CALCIFICATIONS IN HEMODIALYSIS PATIENTS. INTRODUCTION: THE PREVALENCE OF INTRACRANIAL ARTERIAL CALCIFICATION (ICAC) IN MAINTENANCE HEMODIALYSIS (MHD) PATIENTS IS ABOUT 90%, AND ITS SEVERITY IS CORRELATED WITH AGE, HEMODIALYSIS VINTAGE, AND MINERAL BONE DISEASE. ELEVATED CONCENTRATIONS OF CALCIUM AND PHOSPHORUS ARE NOT SUFFICIENT FOR MEDIAL CALCIFICATION BECAUSE OF INHIBITION BY PYROPHOSPHATE. ALKALINE PHOSPHATASE (ALP) PROMOTES CALCIFICATION BY HYDROLYZING EXTRACELLULAR PYROPHOSPHATE. EPIGENETIC MECHANISMS INVOLVING ALP INHIBITION BY APABETALONE WERE INVESTIGATED AS A POTENTIAL TARGET FOR PREVENTING VASCULAR CALCIFICATIONS (VCS). THIS STUDY ASSESSED THE COMBINED IMPACT OF VCS AND ELEVATED SERUM ALP ON MORTALITY AMONG CHRONIC HD PATIENTS. METHODS: VCS REPRESENTED BY ICAC WERE MEASURED SIMULTANEOUSLY WITH MINERAL BONE DISEASE PARAMETERS INCLUDING SERUM ALP OF MHD PATIENTS WHO UNDERWENT NONCONTRAST BRAIN COMPUTED TOMOGRAPHY FROM 2015 TO 2018 IN OUR INSTITUTION. RESULTS: THIS RETROSPECTIVE STUDY INCLUDED 150 MHD PATIENTS (MEAN AGE 71.3 +/- 12.1 YEARS, 60.1% MALE). OF THE TOTAL COHORT, 12 (7.8%) HAD NO BRAIN CALCIFICATIONS AND 69 (45.1%) HAD MULTIPLE INTRACRANIAL CALCIFICATIONS. CONSIDERING THE PATIENTS WITH NORMAL ALP AND NO CALCIFICATION AS THE REFERENCE GROUP YIELDED ADJUSTED ODDS RATIOS FOR ALL-CAUSE MORTALITY OF 4.6 (95% CI: 1.7-12.7) AMONG PATIENTS WITH BRAIN CALCIFICATIONS AND NORMAL ALP (P = 0.003) AND ODDS RATIOS FOR ALL-CAUSE MORTALITY OF 6.1 (95% CI: 2.1-17.7) AMONG PATIENTS WITH BRAIN CALCIFICATIONS AND ELEVATED ALP (P= 0.001). CONCLUSION: WE FOUND AN INDEPENDENT ASSOCIATION BETWEEN ICAC AND THE RISK OF DEATH AMONG MHD PATIENTS. THE COMBINED EFFECT OF ICAC AND ELEVATED ALP WAS ASSOCIATED WITH A HIGHER ODDS RATIO FOR ALL-CAUSE MORTALITY IN MHD PATIENTS AND MAY CONTRIBUTE TO THE RISK STRATIFICATION OF THESE PATIENTS.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
4  1785  52 EFFECT OF APABETALONE ON CARDIOVASCULAR EVENTS IN DIABETES, CKD, AND RECENT ACUTE CORONARY SYNDROME: RESULTS FROM THE BETONMACE RANDOMIZED CONTROLLED TRIAL. BACKGROUND AND OBJECTIVES: CKD AND TYPE 2 DIABETES MELLITUS INTERACT TO INCREASE THE RISK OF MAJOR ADVERSE CARDIOVASCULAR EVENTS (I.E., CARDIOVASCULAR DEATH, NONFATAL MYOCARDIAL INFARCTION, OR STROKE) AND CONGESTIVE HEART FAILURE. A MALADAPTIVE EPIGENETIC RESPONSE MAY BE A CARDIOVASCULAR RISK DRIVER AND AMENABLE TO MODIFICATION WITH APABETALONE, A SELECTIVE MODULATOR OF THE BROMODOMAIN AND EXTRATERMINAL DOMAIN TRANSCRIPTION SYSTEM. WE EXAMINED THIS QUESTION IN A PRESPECIFIED ANALYSIS OF BETONMACE, A PHASE 3 TRIAL. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: BETONMACE WAS AN EVENT-DRIVEN, RANDOMIZED, DOUBLE-BLIND, PLACEBO-CONTROLLED TRIAL COMPARING EFFECTS OF APABETALONE VERSUS PLACEBO ON MAJOR ADVERSE CARDIOVASCULAR EVENTS AND HEART FAILURE HOSPITALIZATIONS IN 2425 PARTICIPANTS WITH TYPE 2 DIABETES AND A RECENT ACUTE CORONARY SYNDROME, INCLUDING 288 PARTICIPANTS WITH CKD WITH EGFR <60 ML/MIN PER 1.73 M(2) AT BASELINE. THE PRIMARY END POINT IN BETONMACE WAS THE TIME TO THE FIRST MAJOR ADVERSE CARDIOVASCULAR EVENT, WITH A SECONDARY END POINT OF TIME TO HOSPITALIZATION FOR HEART FAILURE. RESULTS: MEDIAN FOLLOW-UP WAS 27 MONTHS (INTERQUARTILE RANGE, 20-32 MONTHS). IN PARTICIPANTS WITH CKD, APABETALONE COMPARED WITH PLACEBO WAS ASSOCIATED WITH FEWER MAJOR ADVERSE CARDIOVASCULAR EVENTS (13 EVENTS IN 124 PATIENTS [11%] VERSUS 35 EVENTS IN 164 PATIENTS [21%]; HAZARD RATIO, 0.50; 95% CONFIDENCE INTERVAL, 0.26 TO 0.96) AND FEWER HEART FAILURE-RELATED HOSPITALIZATIONS (THREE HOSPITALIZATIONS IN 124 PATIENTS [3%] VERSUS 14 HOSPITALIZATIONS IN 164 PATIENTS [9%]; HAZARD RATIO, 0.48; 95% CONFIDENCE INTERVAL, 0.26 TO 0.86). IN THE NON-CKD GROUP, THE CORRESPONDING HAZARD RATIO VALUES WERE 0.96 (95% CONFIDENCE INTERVAL, 0.74 TO 1.24) FOR MAJOR ADVERSE CARDIOVASCULAR EVENTS, AND 0.76 (95% CONFIDENCE INTERVAL, 0.46 TO 1.27) FOR HEART FAILURE-RELATED HOSPITALIZATION. INTERACTION OF CKD ON TREATMENT EFFECT WAS P=0.03 FOR MAJOR ADVERSE CARDIOVASCULAR EVENTS, AND P=0.12 FOR HEART FAILURE-RELATED HOSPITALIZATION. PARTICIPANTS WITH CKD SHOWED SIMILAR NUMBERS OF ADVERSE EVENTS, REGARDLESS OF RANDOMIZATION TO APABETALONE OR PLACEBO (119 [73%] VERSUS 88 [71%] PATIENTS), AND THERE WERE FEWER SERIOUS ADVERSE EVENTS (29% VERSUS 43%; P=0.02) IN THE APABETALONE GROUP. CONCLUSIONS: APABETALONE MAY REDUCE THE INCIDENCE OF MAJOR ADVERSE CARDIOVASCULAR EVENTS IN PATIENTS WITH CKD AND TYPE 2 DIABETES WHO HAVE A HIGH BURDEN OF CARDIOVASCULAR DISEASE.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                   
5   446  37 APABETALONE DOWNREGULATES FIBROTIC, INFLAMMATORY AND CALCIFIC PROCESSES IN RENAL MESANGIAL CELLS AND PATIENTS WITH RENAL IMPAIRMENT. EPIGENETIC MECHANISMS ARE IMPLICATED IN TRANSCRIPTIONAL PROGRAMS DRIVING CHRONIC KIDNEY DISEASE (CKD). APABETALONE IS AN ORALLY AVAILABLE INHIBITOR OF BROMODOMAIN AND EXTRATERMINAL (BET) PROTEINS, WHICH ARE EPIGENETIC READERS THAT MODULATE GENE EXPRESSION. IN THE PHASE 3 BETONMACE TRIAL, APABETALONE REDUCED RISK OF MAJOR ADVERSE CARDIAC EVENTS (MACE) BY 50% IN THE CKD SUBPOPULATION, INDICATING FAVORABLE EFFECTS ALONG THE KIDNEY-HEART AXIS. ACTIVATION OF HUMAN RENAL MESANGIAL CELLS (HRMCS) TO A CONTRACTILE PHENOTYPE THAT OVERPRODUCES EXTRACELLULAR MATRIX (ECM) AND INFLAMMATORY CYTOKINES, AND PROMOTES CALCIFICATION, FREQUENTLY ACCOMPANIES CKD TO DRIVE PATHOLOGY. HERE, WE SHOW APABETALONE DOWNREGULATED HRMC ACTIVATION WITH TGF-BETA1 STIMULATION BY SUPPRESSING TGF-BETA1-INDUCED ALPHA-SMOOTH MUSCLE ACTIN (ALPHA-SMA) EXPRESSION, ALPHA-SMA ASSEMBLY INTO STRESS FIBERS, ENHANCED CONTRACTION, COLLAGEN OVERPRODUCTION, AND EXPRESSION OF KEY DRIVERS OF FIBROSIS, INFLAMMATION, OR CALCIFICATION INCLUDING THROMBOSPONDIN, FIBRONECTIN, PERIOSTIN, SPARC, INTERLEUKIN 6, AND ALKALINE PHOSPHATASE. LIPOPOLYSACCHARIDE-STIMULATED EXPRESSION OF INFLAMMATORY GENES IL6, IL1B, AND PTGS2 WAS ALSO SUPPRESSED. TRANSCRIPTOMICS CONFIRMED APABETALONE AFFECTED GENE SETS OF ECM REMODELING AND INTEGRINS. CLINICAL TRANSLATION OF IN VITRO RESULTS WAS INDICATED IN CKD PATIENTS WHERE A SINGLE DOSE OF APABETALONE REDUCED PLASMA LEVELS OF KEY PRO-FIBROTIC AND INFLAMMATORY MARKERS, AND INDICATED INHIBITION OF TGF-BETA1 SIGNALING. WHILE PLASMA PROTEINS CANNOT BE TRACED TO THE KIDNEY ALONE, ANTI-FIBROTIC AND ANTI-INFLAMMATORY EFFECTS OF APABETALONE IDENTIFIED IN THIS STUDY ARE CONSISTENT WITH THE OBSERVED DECREASE IN CARDIOVASCULAR RISK IN CKD PATIENTS.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
6  5045  36 PHARMACOLOGIC EPIGENETIC MODULATORS OF ALKALINE PHOSPHATASE IN CHRONIC KIDNEY DISEASE. PURPOSE OF REVIEW: IN CHRONIC KIDNEY DISEASE (CKD), DISTURBANCE OF SEVERAL METABOLIC REGULATORY MECHANISMS CAUSE PREMATURE AGEING, ACCELERATED CARDIOVASCULAR DISEASE (CVD), AND MORTALITY. SINGLE-TARGET INTERVENTIONS HAVE REPEATEDLY FAILED TO IMPROVE THE PROGNOSIS FOR CKD PATIENTS. EPIGENETIC INTERVENTIONS HAVE THE POTENTIAL TO MODULATE SEVERAL PATHOGENETIC PROCESSES SIMULTANEOUSLY. ALKALINE PHOSPHATASE (ALP) IS A ROBUST PREDICTOR OF CVD AND ALL-CAUSE MORTALITY AND IMPLICATED IN PATHOGENIC PROCESSES ASSOCIATED WITH CVD IN CKD. RECENT FINDINGS: IN EXPERIMENTAL STUDIES, EPIGENETIC MODULATION OF ALP BY MICRORNAS OR BROMODOMAIN AND EXTRATERMINAL (BET) PROTEIN INHIBITION HAS SHOWN PROMISING RESULTS FOR THE TREATMENT OF CVD AND OTHER CHRONIC METABOLIC DISEASES. THE BET INHIBITOR APABETALONE IS CURRENTLY BEING EVALUATED FOR CARDIOVASCULAR RISK REDUCTION IN A PHASE III CLINICAL STUDY IN HIGH-RISK CVD PATIENTS, INCLUDING PATIENTS WITH CKD (CLINICALTRIALS.GOV IDENTIFIER: NCT02586155). PHASE II STUDIES DEMONSTRATE AN ALP-LOWERING POTENTIAL OF APABETALONE, WHICH WAS ASSOCIATED WITH IMPROVED CARDIOVASCULAR AND RENAL OUTCOMES. SUMMARY: ALP IS A PREDICTOR OF CVD AND MORTALITY IN CKD. EPIGENETIC MODULATION OF ALP HAS THE POTENTIAL TO AFFECT SEVERAL PATHOGENETIC PROCESSES IN CKD AND THEREBY IMPROVE CARDIOVASCULAR OUTCOME.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
7   445  44 APABETALONE (RVX-208) REDUCES VASCULAR INFLAMMATION IN VITRO AND IN CVD PATIENTS BY A BET-DEPENDENT EPIGENETIC MECHANISM. BACKGROUND: APABETALONE (RVX-208) IS A BROMODOMAIN AND EXTRATERMINAL PROTEIN INHIBITOR (BETI) THAT IN PHASE II TRIALS REDUCED THE RELATIVE RISK (RR) OF MAJOR ADVERSE CARDIAC EVENTS (MACE) IN PATIENTS WITH CARDIOVASCULAR DISEASE (CVD) BY 44% AND IN DIABETIC CVD PATIENTS BY 57% ON TOP OF STATINS. A PHASE III TRIAL, BETONMACE, IS CURRENTLY ASSESSING APABETALONE'S ABILITY TO REDUCE MACE IN STATIN-TREATED POST-ACUTE CORONARY SYNDROME TYPE 2 DIABETIC CVD PATIENTS WITH LOW HIGH-DENSITY LIPOPROTEIN C. THE LEADING CAUSE OF MACE IS ATHEROSCLEROSIS, DRIVEN BY DYSFUNCTIONAL LIPID METABOLISM AND CHRONIC VASCULAR INFLAMMATION (VI). IN VITRO STUDIES HAVE IMPLICATED THE BET PROTEIN BRD4 AS AN EPIGENETIC DRIVER OF INFLAMMATION AND ATHEROGENESIS, SUGGESTING THAT BETI MAY BE CLINICALLY EFFECTIVE IN COMBATING VI. HERE, WE ASSESSED APABETALONE'S ABILITY TO REGULATE INFLAMMATION-DRIVEN GENE EXPRESSION AND CELL ADHESION IN VITRO AND INVESTIGATED THE MECHANISM BY WHICH APABETALONE SUPPRESSES EXPRESSION. THE CLINICAL IMPACT OF APABETALONE ON MEDIATORS OF VI WAS ASSESSED WITH PROTEOMIC ANALYSIS OF PHASE II CVD PATIENT PLASMA. RESULTS: IN VITRO, APABETALONE PREVENTED INFLAMMATORY (TNFALPHA, LPS, OR IL-1BETA) INDUCTION OF KEY FACTORS THAT DRIVE ENDOTHELIAL ACTIVATION, MONOCYTE RECRUITMENT, ADHESION, AND PLAQUE DESTABILIZATION. BRD4 ABUNDANCE ON INFLAMMATORY AND ADHESION GENE PROMOTERS AND ENHANCERS WAS REDUCED BY APABETALONE. BRD2-4 DEGRADATION BY MZ-1 ALSO PREVENTED TNFALPHA-INDUCED TRANSCRIPTION OF MONOCYTE AND ENDOTHELIAL CELL ADHESION MOLECULES AND INFLAMMATORY MEDIATORS, CONFIRMING BET-DEPENDENT REGULATION. TRANSCRIPTIONAL REGULATION BY APABETALONE TRANSLATED INTO A REDUCTION IN MONOCYTE ADHESION TO AN ENDOTHELIAL MONOLAYER. IN A PHASE II TRIAL, APABETALONE TREATMENT REDUCED THE ABUNDANCE OF MULTIPLE VI MEDIATORS IN THE PLASMA OF CVD PATIENTS (SOMASCAN(R) 1.3 K). THESE PROTEINS CORRELATE WITH CVD RISK AND INCLUDE ADHESION MOLECULES, CYTOKINES, AND METALLOPROTEINASES. INGENUITY(R) PATHWAY ANALYSIS (IPA(R)) PREDICTED THAT APABETALONE INHIBITS PRO-ATHEROGENIC REGULATORS AND PATHWAYS AND PREVENTS DISEASE STATES ARISING FROM LEUKOCYTE RECRUITMENT. CONCLUSIONS: APABETALONE SUPPRESSED GENE EXPRESSION OF VI MEDIATORS IN MONOCYTES AND ENDOTHELIAL CELLS BY INHIBITING BET-DEPENDENT TRANSCRIPTION INDUCED BY MULTIPLE INFLAMMATORY STIMULI. IN CVD PATIENTS, APABETALONE TREATMENT REDUCED CIRCULATING LEVELS OF VI MEDIATORS, AN OUTCOME CONDUCIVE WITH ATHEROSCLEROTIC PLAQUE STABILIZATION AND MACE REDUCTION. INHIBITION OF INFLAMMATORY AND ADHESION MOLECULE GENE EXPRESSION BY APABETALONE IS PREDICTED TO CONTRIBUTE TO MACE REDUCTION IN THE PHASE III BETONMACE TRIAL.	2019	
                                                                                                                                                                                                                                                                  
8   322  32 ALKALINE PHOSPHATASE: AN OLD FRIEND AS TREATMENT TARGET FOR CARDIOVASCULAR AND MINERAL BONE DISORDERS IN CHRONIC KIDNEY DISEASE. ALKALINE PHOSPHATASE (ALP) IS AN EVOLUTIONARY CONSERVED ENZYME AND WIDELY USED BIOMARKER IN CLINICAL PRACTICE. TISSUE-NONSPECIFIC ALKALINE PHOSPHATASE (TNALP) IS ONE OF FOUR HUMAN ISOZYMES THAT ARE EXPRESSED AS DISTINCT TNALP ISOFORMS AFTER POSTTRANSLATIONAL MODIFICATIONS, MAINLY IN BONE, LIVER, AND KIDNEY TISSUES. BEYOND THE WELL-KNOWN EFFECTS ON BONE MINERALIZATION, THE BONE ALP (BALP) ISOFORMS (B/I, B1, B1X, AND B2) ARE ALSO INVOLVED IN THE PATHOGENESIS OF ECTOPIC CALCIFICATION. THIS NARRATIVE REVIEW SUMMARIZES THE RECENT CLINICAL INVESTIGATIONS AND MECHANISMS THAT LINK ALP AND BALP TO INFLAMMATION, METABOLIC SYNDROME, VASCULAR CALCIFICATION, ENDOTHELIAL DYSFUNCTION, FIBROSIS, CARDIOVASCULAR DISEASE, AND MORTALITY. THE ASSOCIATION BETWEEN ALP, VITAMIN K, BONE METABOLISM, AND FRACTURE RISK IN PATIENTS WITH CHRONIC KIDNEY DISEASE (CKD) IS ALSO DISCUSSED. RECENT ADVANCES IN DIFFERENT PHARMACOLOGICAL STRATEGIES ARE HIGHLIGHTED, WITH THE POTENTIAL TO MODULATE THE EXPRESSION OF ALP DIRECTLY AND INDIRECTLY IN CKD-MINERAL AND BONE DISORDER (CKD-MBD), E.G., EPIGENETIC MODULATION, PHOSPHATE BINDERS, CALCIMIMETICS, VITAMIN D, AND OTHER ANTI-FRACTURE TREATMENTS. WE CONCLUDE THAT THE SIGNIFICANT EVIDENCE FOR ALP AS A PATHOGENIC FACTOR AND RISK MARKER IN CKD-MBD SUPPORTS THE INCLUSION OF CONCRETE TREATMENT TARGETS FOR ALP IN CLINICAL GUIDELINES. WHILE A TARGET VALUE BELOW 120 U/L IS ASSOCIATED WITH IMPROVED SURVIVAL, FURTHER EXPERIMENTAL AND CLINICAL RESEARCH SHOULD EXPLORE INTERVENTIONAL STRATEGIES WITH OPTIMAL RISK-BENEFIT PROFILES. THE FUTURE HOLDS GREAT PROMISE FOR NOVEL DRUG THERAPIES MODULATING ALP.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
9   974  26 CHRONIC OCCUPATIONAL EXPOSURE ENDURED BY TOBACCO FARMERS FROM BRAZIL AND ASSOCIATION WITH DNA DAMAGE. TOBACCO FARMING IS AN IMPORTANT ECONOMIC INCOME IN BRAZIL, ALTHOUGH IT HAS BEEN CHALLENGED AS REGARD THE OCCUPATIONAL EXPOSURE TO BOTH PESTICIDES AND NICOTINE ENDURED BY FARMERS. CHRONIC OCCUPATIONAL EXPOSURE TO COMPLEX MIXTURES CAN LEAD TO HEALTH HAZARDOUS. WE EXAMINED GENOMIC INSTABILITY AND EPIGENETIC CHANGES IN TOBACCO FARMERS OCCUPATIONALLY EXPOSED TO PESTICIDE MIXTURES AND NICOTINE AT TOBACCO FIELDS. DNA DAMAGE WAS ASSESSED BY ALKALINE COMET ASSAY IN BLOOD CELLS. GENOMIC DNA WAS ISOLATED, AND TELOMERE LENGTH WAS MEASURED USING QUANTITATIVE POLYMERASE CHAIN REACTION ASSAY. WE MEASURED 5-METHYL-2'-DEOXYCYTIDINE, A MARKER OF GLOBAL DNA METHYLATION, AND P16 PROMOTER METHYLATION. THE OXIDATIVE PROFILE WAS EVALUATED BY TROLOX EQUIVALENT ANTIOXIDANT CAPACITY AND LIPID PEROXIDATION (THIOBARBITURIC ACID REACTIVE SUBSTANCES) IN SERUM. EXPOSURE PARAMETERS, PLASMA COTININE AND INORGANIC ELEMENT LEVELS, WERE ALSO MEASURED. DNA DAMAGE WAS SIGNIFICANTLY ELEVATED FOR FARMERS IN RELATION TO UNEXPOSED GROUP (P < 0.001; MANN-WHITNEY TEST) AND POSITIVELY ASSOCIATED WITH YEARS OF EXPOSURE. INVERSE RELATIONSHIP BETWEEN DNA DAMAGE AND TOTAL EQUIVALENT ANTIOXIDANT ACTIVITY WAS DEMONSTRATED FOR EXPOSED AND UNEXPOSED GROUPS. EXPOSED GROUP SHOWED SIGNIFICANTLY SHORTER TELOMERES (P < 0.001; UNPAIRED T-TEST) AND DNA HYPOMETHYLATION (P < 0.001; UNPAIRED T-TEST), AS WELL AS P16 HYPERMETHYLATION (P = 0.003; MANN-WHITNEY TEST). LIPID PEROXIDATION WAS INCREASED FOR EXPOSED GROUP IN RELATION TO UNEXPOSED ONE (P = 0.02; MANN-WHITNEY TEST) AND PRESENTED A POSITIVE CORRELATION WITH GLOBAL DNA METHYLATION (P = 0.0264). FARMERS HAVE INCREASED PLASMA COTININE LEVELS (P < 0.001) AND INORGANIC ELEMENTS (PHOSPHORUS, SULPHUR AND CHLORINE) IN RELATION TO UNEXPOSED GROUP. ELEVATED OXIDATIVE STRESS LEVELS DUE TO CHRONIC OCCUPATIONAL PESTICIDE MIXTURES AND NICOTINE EXPOSURE IN TOBACCO FARMERS WERE ASSOCIATED WITH HIGHER DNA DAMAGE, SHORTER TELOMERES AND ALTERED DNA METHYLATION. TELOMERE-ACCELERATED ATTRITION DUE TO EXPOSURE MAY BE POTENTIAL INTERMEDIATE STEP BEFORE A DISEASE STATE.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
10  2239  32 EPIGENETIC MODULATION BY APABETALONE COUNTERS CYTOKINE-DRIVEN ACUTE PHASE RESPONSE IN VITRO, IN MICE AND IN PATIENTS WITH CARDIOVASCULAR DISEASE. CHRONIC SYSTEMIC INFLAMMATION CONTRIBUTES TO CARDIOVASCULAR DISEASE (CVD) AND CORRELATES WITH THE ABUNDANCE OF ACUTE PHASE RESPONSE (APR) PROTEINS IN THE LIVER AND PLASMA. BROMODOMAIN AND EXTRATERMINAL (BET) PROTEINS ARE EPIGENETIC READERS THAT REGULATE INFLAMMATORY GENE TRANSCRIPTION. WE SHOW THAT BET INHIBITION BY THE SMALL MOLECULE APABETALONE REDUCES APR GENE AND PROTEIN EXPRESSION IN HUMAN HEPATOCYTES, MOUSE MODELS, AND PLASMA FROM CVD PATIENTS. STEADY-STATE EXPRESSION OF SERUM AMYLOID P, PLASMINOGEN ACTIVATOR INHIBITOR 1, AND CERULOPLASMIN, APR PROTEINS LINKED TO CVD RISK, IS REDUCED BY APABETALONE IN CULTURED HEPATOCYTES AND IN HUMANIZED MOUSE LIVER. IN CYTOKINE-STIMULATED HEPATOCYTES, APABETALONE REDUCES THE EXPRESSION OF C-REACTIVE PROTEIN (CRP), ALPHA-2-MACROGLOBULIN, AND SERUM AMYLOID P. THE LATTER TWO ARE ALSO REDUCED BY APABETALONE IN THE LIVER OF ENDOTOXEMIC MICE. BET KNOCKDOWN IN VITRO ALSO COUNTERS CYTOKINE-MEDIATED INDUCTION OF THE CRP GENE. MECHANISTICALLY, APABETALONE REDUCES THE CYTOKINE-DRIVEN INCREASE IN BRD4 BET OCCUPANCY AT THE CRP PROMOTER, CONFIRMING THAT TRANSCRIPTION OF CRP IS BET-DEPENDENT. IN PATIENTS WITH STABLE CORONARY DISEASE, PLASMA APR PROTEINS CRP, IL-1 RECEPTOR ANTAGONIST, AND FIBRINOGEN GAMMA DECREASE AFTER APABETALONE TREATMENT VERSUS PLACEBO, RESULTING IN A PREDICTED DOWNREGULATION OF THE APR PATHWAY AND CYTOKINE TARGETS. WE CONCLUDE THAT CRP AND COMPONENTS OF THE APR PATHWAY ARE REGULATED BY BET PROTEINS AND THAT APABETALONE COUNTERS CHRONIC CYTOKINE SIGNALING IN PATIENTS.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
11  1391  24 DIAGNOSTIC APPROACH TO PATIENTS WITH LOW SERUM ALKALINE PHOSPHATASE. INCREASED SERUM LEVELS OF ALKALINE PHOSPHATASE (ALP) ARE WIDELY RECOGNIZED AS A BIOCHEMICAL MARKER OF MANY DISORDERS AFFECTING THE LIVER OR BONE. HOWEVER, THE APPROACH FOR PATIENTS WITH LOW ALP PHOSPHATASE IS NOT WELL-ESTABLISHED. LOW SERUM ALP IS AN EPIPHENOMENON OF MANY SEVERE ACUTE INJURIES AND DISEASES. PERSISTENTLY LOW SERUM ALP MAY BE SECONDARY TO DRUG THERAPY (INCLUDING ANTIRESORPTIVES) OR A VARIETY OF ACQUIRED DISORDERS, SUCH AS MALNUTRITION, VITAMIN AND MINERAL DEFICIENCIES, ENDOCRINE DISORDERS, ETC. HYPOPHOSPHATASIA, DUE TO PATHOGENIC VARIANTS OF THE ALPL GENE, WHICH ENCODES TISSUE NON-SPECIFIC ALP, IS THE MOST COMMON GENETIC CAUSE OF LOW SERUM ALP. MARKED BONE HYPOMINERALIZATION IS FREQUENT IN SEVERE PEDIATRIC-ONSET CASES. HOWEVER, ADULT FORMS OF HYPOPHOSPHATASIA USUALLY PRESENT WITH MILDER MANIFESTATIONS, SUCH AS SKELETAL PAIN, CHONDROCALCINOSIS, CALCIFIC PERIARTHRITIS, DENTAL PROBLEMS, AND STRESS FRACTURES. THE DIAGNOSTIC APPROACH TO THESE PATIENTS IS DISCUSSED. MEASURING SEVERAL ALP SUBSTRATES, SUCH AS PYROPHOSPHATE, PYRIDOXAL PHOSPHATE, OR PHOSPHOETHANOLAMINE, MAY HELP TO ESTABLISH ENZYME DEFICIENCY. GENE ANALYSIS SHOWING A PATHOGENIC VARIANT IN ALPL MAY CONFIRM THE DIAGNOSIS. HOWEVER, A SUBSTANTIAL PROPORTION OF PATIENTS SHOW NORMAL RESULTS AFTER SEQUENCING ALPL EXONS. IT IS STILL UNKNOWN IF THOSE PATIENTS CARRY UNIDENTIFIED MUTATIONS IN REGULATORY REGIONS OF ALPL, EPIGENETIC CHANGES, OR ABNORMALITIES IN OTHER GENES.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
12   587  43 BENEFIT OF APABETALONE ON PLASMA PROTEINS IN RENAL DISEASE. INTRODUCTION: APABETALONE, A SMALL MOLECULE INHIBITOR, TARGETS EPIGENETIC READERS TERMED BET PROTEINS THAT CONTRIBUTE TO GENE DYSREGULATION IN HUMAN DISORDERS. APABETALONE HAS IN VITRO AND IN VIVO ANTI-INFLAMMATORY AND ANTIATHEROSCLEROTIC PROPERTIES. IN PHASE 2 CLINICAL TRIALS, THIS DRUG REDUCED THE INCIDENCE OF MAJOR ADVERSE CARDIAC EVENTS IN PATIENTS WITH CARDIOVASCULAR DISEASE. CHRONIC KIDNEY DISEASE IS ASSOCIATED WITH A PROGRESSIVE LOSS OF RENAL FUNCTION AND A HIGH RISK OF CARDIOVASCULAR DISEASE. WE STUDIED THE IMPACT OF APABETALONE ON THE PLASMA PROTEOME IN PATIENTS WITH IMPAIRED KIDNEY FUNCTION. METHODS: SUBJECTS WITH STAGE 4 OR 5 CHRONIC KIDNEY DISEASE AND MATCHED CONTROLS RECEIVED A SINGLE DOSE OF APABETALONE. PLASMA WAS COLLECTED FOR PHARMACOKINETIC ANALYSIS AND FOR PROTEOMICS PROFILING USING THE SOMASCAN 1.3K PLATFORM. PROTEOMICS DATA WERE ANALYZED WITH INGENUITY PATHWAY ANALYSIS TO IDENTIFY DYSREGULATED PATHWAYS IN DISEASED PATIENTS, WHICH WERE TARGETED BY APABETALONE. RESULTS: AT BASELINE, 169 PLASMA PROTEINS (ADJUSTED P VALUE <0.05) WERE DIFFERENTIALLY ENRICHED IN RENALLY IMPAIRED PATIENTS VERSUS CONTROL SUBJECTS, INCLUDING CYSTATIN C AND BETA(2) MICROGLOBULIN, WHICH CORRELATE WITH RENAL FUNCTION. BIOINFORMATICS ANALYSIS OF THE PLASMA PROTEOME REVEALED A SIGNIFICANT ACTIVATION OF 42 PATHWAYS THAT CONTROL IMMUNITY AND INFLAMMATION, OXIDATIVE STRESS, ENDOTHELIAL DYSFUNCTION, VASCULAR CALCIFICATION, AND COAGULATION. AT 12 HOURS POSTDOSE, APABETALONE COUNTERED THE ACTIVATION OF PATHWAYS ASSOCIATED WITH RENAL DISEASE AND REDUCED THE ABUNDANCE OF DISEASE MARKERS, INCLUDING INTERLEUKIN-6, PLASMINOGEN ACTIVATOR INHIBITOR-1, AND OSTEOPONTIN. CONCLUSION: THESE DATA DEMONSTRATED PLASMA PROTEOME DYSREGULATION IN RENALLY IMPAIRED PATIENTS AND THE BENEFICIAL IMPACT OF APABETALONE ON PATHWAYS LINKED TO CHRONIC KIDNEY DISEASE AND ITS CARDIOVASCULAR COMPLICATIONS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
13  3295  32 HIGH PHOSPHATE-INDUCED DOWNREGULATION OF PPARGAMMA CONTRIBUTES TO CKD-ASSOCIATED VASCULAR CALCIFICATION. MEDIAL ARTERIAL CALCIFICATION ASSOCIATED WITH HYPERPHOSPHATEMIA IS A MAIN CAUSE OF CARDIOVASCULAR MORTALITY IN PATIENTS WITH CHRONIC KIDNEY DISEASE (CKD), BUT THE MECHANISMS UNDERLYING HIGH PHOSPHATE-INDUCED VASCULAR CALCIFICATION REMAIN LARGELY UNKNOWN. HERE, WE OBSERVED A SIGNIFICANT DECREASE IN THE EXPRESSION OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-GAMMA (PPARGAMMA) IN CALCIFIED ARTERIES BOTH IN CKD PATIENTS AND IN A MOUSE MODEL OF CKD WITH HYPERPHOSPHATEMIA. IN VITRO, HIGH PHOSPHATE TREATMENT LED TO A DECREASED EXPRESSION OF PPARGAMMA IN MOUSE VASCULAR SMOOTH MUSCLE CELLS (VMSCS), ACCOMPANIED BY APPARENT OSTEOGENIC DIFFERENTIATION AND CALCIFICATION. PRETREATMENT WITH PPARGAMMA AGONIST ROSIGLITAZONE SIGNIFICANTLY REVERSED HIGH PHOSPHATE-INDUCED VSMCS CALCIFICATION. FURTHER INVESTIGATION SHOWED THAT METHYL-CPG BINDING PROTEIN 2 (MECP2)-MEDIATED EPIGENETIC REPRESSION WAS INVOLVED IN HIGH PHOSPHATE-INDUCED PPARGAMMA DOWNREGULATION. MOREOVER, THE EXPRESSION OF KLOTHO THAT HAS THE ABILITY TO INHIBIT VASCULAR CALCIFICATION BY REGULATING PHOSPHATE UPTAKE DECREASED WITH THE PPARGAMMA REDUCTION IN VSMCS AFTER HIGH PHOSPHATE TREATMENT, AND ROSIGLITAZONE FAILED TO INHIBIT HIGH PHOSPHATE-INDUCED CALCIFICATION IN VSMCS WITH KNOCKDOWN OF KLOTHO OR IN AORTIC RINGS FROM KLOTHO-DEFICIENT (KL/KL) MICE. FINALLY, AN IN VIVO STUDY DEMONSTRATED THAT ORAL ADMINISTRATION OF ROSIGLITAZONE COULD INCREASE KLOTHO EXPRESSION AND PROTECT AGAINST HIGH PHOSPHATE-INDUCED VASCULAR CALCIFICATION IN CKD MICE. THESE FINDINGS SUGGEST THAT THE INHIBITION OF PPARGAMMA EXPRESSION MAY CONTRIBUTE TO THE PATHOGENESIS OF HIGH PHOSPHATE-INDUCED VASCULAR CALCIFICATION, WHICH MAY PROVIDE A NEW THERAPEUTIC TARGET FOR VASCULAR CALCIFICATION IN CKD PATIENTS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
14   364  40 AMELIORATION OF UREMIC TOXIN INDOXYL SULFATE-INDUCED OSTEOBLASTIC CALCIFICATION BY SET DOMAIN CONTAINING LYSINE METHYLTRANSFERASE 7/9 PROTEIN. BACKGROUND: VASCULAR CALCIFICATION (VC) IS A VERY COMMON PHENOMENON IN PATIENTS WITH CHRONIC KIDNEY DISEASE (CKD). IT HAS BEEN REPORTED THAT SOME HISTONE METHYLATION PLAY A ROLE IN VC AS AN EPIGENETIC REGULATOR. INDOXYL SULFATE (IS) IS A PROTEIN-BOUND UREMIC TOXIN THAT HAS BEEN PROVEN AS ONE OF THE MAJOR RISK FACTORS OF CARDIOVASCULAR DISEASE IN CKD. SET DOMAIN CONTAINING LYSINE METHYLTRANSFERASE 7/9 (SET7/9) IS ONE OF THE IMPORTANT HISTONE METHYLTRANSFERASES. OBJECTIVES: THIS STUDY AIMED TO DETERMINE THE EFFECT OF IS ON THE EXPRESSION OF SET7/9 AND THE ROLE OF SET7/9 IN IS-INDUCED OSTEOBLASTIC DIFFERENTIATION AND CALCIFICATION OF VASCULAR SMOOTH MUSCLE CELLS (VSMCS). METHODS: VSMCS WERE INCUBATED WITH VARIOUS CONCENTRATIONS OF IS FOR DIFFERENT DURATIONS TO ASSESS OSTEOBLASTIC DIFFERENTIATION AND EXPRESSION OF SET7/9. WESTERN BLOT ANALYSIS AND QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION WERE PERFORMED TO ASSESS THE PROTEIN AND MRNA LEVELS OF SET7/9 RESPECTIVELY. THE CALCIUM CONTENT WAS MEASURED TO EVALUATE CALCIFICATION. RESULTS: OSTEOBLASTIC DIFFERENTIATION AND CALCIFICATION OF VSMCS AND DOWNREGULATION OF THE EXPRESSION OF SET7/9 WERE OBSERVED AFTER IS TREATMENT. THE AUTOPHAGY WAS ACTIVATED AFTER TREATMENT WITH IS, WHEREAS THE INHIBITION OF THE AUTOPHAGY PARTIALLY ATTENUATED THE EFFECT OF IS ON BOTH THE STIMULATION OF THE EXPRESSION OF RUNT-RELATED TRANSCRIPTION FACTOR 2 AND CALCIUM DEPOSITION. CONCLUSIONS: OUR DATA DEMONSTRATED THAT SET7/9 DOWNREGULATION AND AUTOPHAGY ACTIVATION MAY BE THE KEY MECHANISM OF IS-INDUCED VC IN CKD.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
15  2130  17 EPIGENETIC INACTIVATION OF THE AUTOPHAGY-LYSOSOMAL SYSTEM IN APPENDIX IN PARKINSON'S DISEASE. THE GASTROINTESTINAL TRACT MAY BE A SITE OF ORIGIN FOR ALPHA-SYNUCLEIN PATHOLOGY IN IDIOPATHIC PARKINSON'S DISEASE (PD). DISRUPTION OF THE AUTOPHAGY-LYSOSOME PATHWAY (ALP) MAY CONTRIBUTE TO ALPHA-SYNUCLEIN AGGREGATION. HERE WE EXAMINED EPIGENETIC ALTERATIONS IN THE ALP IN THE APPENDIX BY DEEP SEQUENCING DNA METHYLATION AT 521 ALP GENES. WE IDENTIFIED ABERRANT METHYLATION AT 928 CYTOSINES AFFECTING 326 ALP GENES IN THE APPENDIX OF INDIVIDUALS WITH PD AND WIDESPREAD HYPERMETHYLATION THAT IS ALSO SEEN IN THE BRAIN OF INDIVIDUALS WITH PD. IN MICE, WE FIND THAT DNA METHYLATION CHANGES AT ALP GENES INDUCED BY CHRONIC GUT INFLAMMATION ARE GREATLY EXACERBATED BY ALPHA-SYNUCLEIN PATHOLOGY. DNA METHYLATION CHANGES AT ALP GENES INDUCED BY SYNUCLEINOPATHY ARE ASSOCIATED WITH THE ALP ABNORMALITIES OBSERVED IN THE APPENDIX OF INDIVIDUALS WITH PD SPECIFICALLY INVOLVING LYSOSOMAL GENES. OUR WORK IDENTIFIES EPIGENETIC DYSREGULATION OF THE ALP WHICH MAY SUGGEST A POTENTIAL MECHANISM FOR ACCUMULATION OF ALPHA-SYNUCLEIN PATHOLOGY IN IDIOPATHIC PD.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
16  1189  36 CORRELATION BETWEEN GLOBAL METHYLATION LEVEL OF PERIPHERAL BLOOD LEUKOCYTES AND SERUM C REACTIVE PROTEIN LEVEL MODIFIED BY MTHFR POLYMORPHISM: A CROSS-SECTIONAL STUDY. BACKGROUND: CHRONIC INFLAMMATORY CONDITIONS ARE ASSOCIATED WITH HIGHER TUMOR INCIDENCE THROUGH EPIGENETIC AND GENETIC ALTERATIONS. HERE, WE FOCUSED ON AN ASSOCIATION BETWEEN AN INFLAMMATION MARKER, C-REACTIVE-PROTEIN (CRP), AND GLOBAL DNA METHYLATION LEVELS OF PERIPHERAL BLOOD LEUKOCYTES. METHODS: THE SUBJECTS WERE 384 HEALTHY JAPANESE WOMEN ENROLLED AS THE CONTROL GROUP OF A CASE-CONTROL STUDY FOR BREAST CANCER CONDUCTED FROM 2001 TO 2005. GLOBAL DNA METHYLATION WAS QUANTIFIED BY LUMINOMETRIC METHYLATION ASSAY (LUMA). RESULTS: WITH ADJUSTMENT FOR LIFESTYLE-RELATED FACTORS, INCLUDING FOLATE INTAKE, THE GLOBAL DNA METHYLATION LEVEL OF PERIPHERAL BLOOD LEUKOCYTES WAS SIGNIFICANTLY BUT WEAKLY INCREASED BY 0.43% PER QUARTILE CATEGORY FOR CRP (P FOR TREND = 0.010). ESTIMATED METHYLATION LEVELS STRATIFIED BY CRP QUARTILE WERE 70.0%, 70.8%, 71.4%, AND 71.3%, RESPECTIVELY. IN ADDITION, INTERACTION BETWEEN POLYMORPHISM OF MTHFR (RS1801133, KNOWN AS C677T) AND CRP WAS SIGNIFICANT (P FOR INTERACTION = 0.046); THE GLOBAL METHYLATION LEVEL WAS SIGNIFICANTLY INCREASED BY 0.61% PER QUARTILE CATEGORY FOR CRP IN THE CT/TT GROUP (THOSE WITH THE MINOR ALLELE T, P FOR TREND = 0.001), WHEREAS NO ASSOCIATION WAS OBSERVED IN THE CC GROUP (WILD TYPE). CONCLUSIONS: OUR STUDY SUGGESTS THAT CRP CONCENTRATION IS WEAKLY ASSOCIATED WITH GLOBAL DNA METHYLATION LEVEL. HOWEVER, THIS ASSOCIATION WAS OBSERVED MORE CLEARLY IN INDIVIDUALS WITH THE MINOR ALLELE OF THE MTHFR MISSENSE SNP RS1801133. BY ELUCIDATING THE COMPLEX MECHANISM OF THE REGULATION OF DNA METHYLATION BY BOTH ACQUIRED AND GENETIC FACTORS, OUR RESULTS MAY BE IMPORTANT FOR CANCER PREVENTION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
17  4249  40 METHYLATION-BASED BIOLOGICAL AGE AND BREAST CANCER RISK. BACKGROUND: AGE IS ONE OF THE STRONGEST PREDICTORS OF CANCER, CHRONIC DISEASE, AND MORTALITY, BUT BIOLOGICAL RESPONSES TO AGING DIFFER AMONG PEOPLE. EPIGENETIC DNA MODIFICATIONS HAVE BEEN USED TO ESTIMATE "BIOLOGICAL AGE," WHICH MAY BE A USEFUL PREDICTOR OF DISEASE RISK. WE TESTED THIS HYPOTHESIS FOR BREAST CANCER. METHODS: USING A CASE-COHORT APPROACH, WE MEASURED BASELINE BLOOD DNA METHYLATION OF 2764 WOMEN ENROLLED IN THE SISTER STUDY, 1566 OF WHOM SUBSEQUENTLY DEVELOPED BREAST CANCER AFTER AN AVERAGE OF 6 YEARS. USING THREE PREVIOUSLY ESTABLISHED METHYLATION-BASED "CLOCKS" (HANNUM, HORVATH, AND LEVINE), WE DEFINED BIOLOGICAL AGE ACCELERATION FOR EACH WOMAN BY COMPARING HER ESTIMATED BIOLOGICAL AGE WITH HER CHRONOLOGICAL AGE. HAZARD RATIOS AND 95% CONFIDENCE INTERVALS FOR BREAST CANCER RISK WERE ESTIMATED USING COX REGRESSION MODELS. ALL STATISTICAL TESTS WERE TWO-SIDED. RESULTS: EACH OF THE THREE CLOCKS SHOWED THAT BIOLOGICAL AGE ACCELERATION WAS STATISTICALLY SIGNIFICANTLY ASSOCIATED WITH INCREASED RISK OF DEVELOPING BREAST CANCER (5-YEAR AGE ACCELERATION, HANNUM'S CLOCK: HAZARD RATIO [HR] = 1.10, 95% CONFIDENCE INTERVAL [CI] = 1.00 TO 1.21, P = .04; HORVATH'S CLOCK: HR = 1.08, 95% CI = 1.00 TO 1.17, P = .04; LEVINE'S CLOCK: HR = 1.15, 95% CI = 1.07 TO 1.23, P < .001). FOR LEVINE'S CLOCK, EACH 5-YEAR ACCELERATION IN BIOLOGICAL AGE CORRESPONDED WITH A 15% INCREASE IN BREAST CANCER RISK. ALTHOUGH BIOLOGICAL AGE MAY ACCELERATE WITH MENOPAUSAL TRANSITION, AGE ACCELERATION IN PREMENOPAUSAL WOMEN INDEPENDENTLY PREDICTED BREAST CANCER. CASE-ONLY ANALYSIS SUGGESTED THAT, AMONG WOMEN WHO DEVELOP BREAST CANCER, INCREASED AGE ACCELERATION IS ASSOCIATED WITH INVASIVE CANCER (ODDS RATIO FOR INVASIVE = 1.09, 95% CI = 0.98 TO 1.22, P = .10). CONCLUSIONS: DNA METHYLATION-BASED MEASURES OF BIOLOGICAL AGE MAY BE IMPORTANT PREDICTORS OF BREAST CANCER RISK.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
18  3623  37 IN VIVO COMET ASSAY ON ISOLATED KIDNEY CELLS TO DISTINGUISH GENOTOXIC CARCINOGENS FROM EPIGENETIC CARCINOGENS OR CYTOTOXIC COMPOUNDS. THE OBJECTIVE OF THIS STUDY WAS TO DETERMINE THE ABILITY OF THE ALKALINE IN VIVO COMET ASSAY (PH>13) TO DISTINGUISH GENOTOXIC CARCINOGENS FROM EPIGENETIC CARCINOGENS WHEN PERFORMED ON FRESHLY ISOLATED KIDNEY CELLS AND TO DETERMINE THE POSSIBLE INTERFERENCE OF CYTOTOXICITY BY ASSESSING DNA DAMAGE INDUCED BY RENAL GENOTOXIC, EPIGENETIC OR TOXIC COMPOUNDS AFTER ENZYMATIC ISOLATION OF KIDNEY CELLS FROM OFA SPRAGUE-DAWLEY MALE RATS. THE ABILITY OF THE COMET ASSAY TO DISTINGUISH (1) GENOTOXICITY VERSUS CYTOTOXICITY AND (2) GENOTOXIC VERSUS NON-GENOTOXIC (EPIGENETIC) CARCINOGENS, WAS THUS INVESTIGATED BY STUDYING FIVE KNOWN GENOTOXIC RENAL CARCINOGENS ACTING THROUGH DIVERSE MECHANISMS OF ACTION, I.E. STREPTOZOTOCIN, ARISTOLOCHIC ACIDS, 2-NITROANISOLE, POTASSIUM BROMATE AND CISPLATIN, TWO RODENT RENAL EPIGENETIC CARCINOGENS: D-LIMONENE AND CICLOSPORINE AND TWO NEPHROTOXIC COMPOUNDS: STREPTOMYCIN AND INDOMETHACIN. ANIMALS WERE TREATED ONCE WITH THE TEST COMPOUND BY THE APPROPRIATE ROUTE OF ADMINISTRATION AND GENOTOXIC EFFECTS WERE MEASURED AT THE TWO SAMPLING TIMES OF 3-6 AND 22-26H AFTER TREATMENT. REGARDING THE TISSUE PROCESSING, THE LIMITED BACKGROUND LEVEL OF DNA MIGRATION OBSERVED IN THE NEGATIVE CONTROL GROUPS THROUGHOUT ALL EXPERIMENTS DEMONSTRATED THAT THE ENZYMATIC ISOLATION METHOD IMPLEMENTED IN THE CURRENT STUDY IS APPROPRIATE. ON THE OTHER HAND, STREPTOZOTOCIN, 20MG/KG, USED AS POSITIVE REFERENCE CONTROL CONCURRENTLY TO EACH ASSAY, CAUSED A CLEAR INCREASE IN THE MEAN OLIVE TAIL MOMENT MEDIAN VALUE, WHICH ALLOWS VALIDATING THE CURRENT METHODOLOGY. UNDER THESE EXPERIMENTAL CONDITIONS, THE IN VIVO RODENT COMET ASSAY DEMONSTRATED GOOD SENSITIVITY AND GOOD SPECIFICITY: ALL THE FIVE RENAL GENOTOXIC CARCINOGENS WERE CLEARLY DETECTED IN AT LEAST ONE EXPRESSION PERIOD EITHER DIRECTLY OR INDIRECTLY, AS IN THE CASE OF CISPLATIN: FOR THIS CROSS-LINKING AGENT, THE SIGNIFICANT DECREASE IN DNA MIGRATION OBSERVED UNDER STANDARD ELECTROPHORESIS CONDITIONS WAS CLEARLY AMPLIFIED WHEN THE DURATION OF ELECTROPHORESIS WAS INCREASED UP TO 40MIN. IN CONTRAST, EPIGENETIC AND NEPHROTOXIC COMPOUNDS FAILED TO INDUCE ANY SIGNIFCANT INCREASE IN DNA MIGRATION. IN CONCLUSION, THE IN VIVO RODENT COMET ASSAY PERFORMED ON ISOLATED KIDNEY CELLS COULD BE USED AS A TOOL TO INVESTIGATE THE GENOTOXIC POTENTIAL OF A TEST COMPOUND IF NEOPLASIC/PRENEOPLASIC CHANGES OCCUR AFTER SUBCHRONIC OR CHRONIC TREATMENTS, IN ORDER TO DETERMINE THE ROLE OF GENOTOXICITY IN TUMOR INDUCTION. MOREOVER, THE EPIGENETIC CARCINOGENS AND CYTOTOXIC COMPOUNDS DISPLAYED CLEARLY NEGATIVE RESPONSES IN THIS STUDY. THESE RESULTS ALLOW EXCLUDING A DNA DIRECT-ACTING MECHANISM OF ACTION AND CAN THUS SUGGEST THAT A THRESHOLD EXISTS. THEREFORE, THE CURRENT IN VIVO RODENT COMET ASSAY COULD CONTRIBUTE TO ELUCIDATE AN EPIGENETIC MECHANISM AND THUS, TO UNDERTAKE A RISK ASSESSMENT ASSOCIATED WITH HUMAN USE, DEPENDING ON THE EXPOSURE LEVEL.	2007	

19  1625  45 DNAM-BASED SIGNATURES OF ACCELERATED AGING AND MORTALITY IN BLOOD ARE ASSOCIATED WITH LOW RENAL FUNCTION. BACKGROUND: THE DIFFERENCE BETWEEN AN INDIVIDUAL'S CHRONOLOGICAL AND DNA METHYLATION PREDICTED AGE (DNAMAGE), TERMED DNAMAGE ACCELERATION (DNAMAA), CAN CAPTURE LIFE-LONG ENVIRONMENTAL EXPOSURES AND AGE-RELATED PHYSIOLOGICAL CHANGES REFLECTED IN METHYLATION STATUS. SEVERAL STUDIES HAVE LINKED DNAMAA TO MORBIDITY AND MORTALITY, YET ITS RELATIONSHIP WITH KIDNEY FUNCTION HAS NOT BEEN ASSESSED. WE EVALUATED THE ASSOCIATIONS BETWEEN SEVEN DNAM AGING AND LIFESPAN PREDICTORS (AS WELL AS GRIMAGE COMPONENTS) AND FIVE KIDNEY TRAITS (ESTIMATED GLOMERULAR FILTRATION RATE [EGFR], URINE ALBUMIN-TO-CREATININE RATIO [UACR], SERUM URATE, MICROALBUMINURIA AND CHRONIC KIDNEY DISEASE [CKD]) IN UP TO 9688 EUROPEAN, AFRICAN AMERICAN AND HISPANIC/LATINO INDIVIDUALS FROM SEVEN POPULATION-BASED STUDIES. RESULTS: WE IDENTIFIED 23 SIGNIFICANT ASSOCIATIONS IN OUR LARGE TRANS-ETHNIC META-ANALYSIS (P < 1.43E-03 AND CONSISTENT DIRECTION OF EFFECT ACROSS STUDIES). AGE ACCELERATION MEASURED BY THE EXTRINSIC AND PHENOAGE ESTIMATORS, AS WELL AS ZHANG'S 10-CPG EPIGENETIC MORTALITY RISK SCORE (MRS), WERE ASSOCIATED WITH ALL PARAMETERS OF POOR KIDNEY HEALTH (LOWER EGFR, PREVALENT CKD, HIGHER UACR, MICROALBUMINURIA AND HIGHER SERUM URATE). SIX OF THESE ASSOCIATIONS WERE INDEPENDENTLY OBSERVED IN EUROPEAN AND AFRICAN AMERICAN POPULATIONS. MRS IN PARTICULAR WAS CONSISTENTLY ASSOCIATED WITH EGFR (BETA = - 0.12, 95% CI = [- 0.16, - 0.08] CHANGE IN LOG-TRANSFORMED EGFR PER UNIT INCREASE IN MRS, P = 4.39E-08), PREVALENT CKD (ODDS RATIO (OR) = 1.78 [1.47, 2.16], P = 2.71E-09) AND HIGHER SERUM URATE LEVELS (BETA = 0.12 [0.07, 0.16], P = 2.08E-06). THE "FIRST-GENERATION" CLOCKS (HANNUM, HORVATH) AND GRIMAGE SHOWED DIFFERENT PATTERNS OF ASSOCIATION WITH THE KIDNEY TRAITS. THREE OF THE DNAM-ESTIMATED COMPONENTS OF GRIMAGE, NAMELY ADRENOMEDULLIN, PLASMINOGEN-ACTIVATION INHIBITION 1 AND PACK YEARS, WERE POSITIVELY ASSOCIATED WITH HIGHER UACR, SERUM URATE AND MICROALBUMINURIA. CONCLUSION: DNAMAGE ACCELERATION AND DNAM MORTALITY PREDICTORS ESTIMATED IN WHOLE BLOOD WERE ASSOCIATED WITH MULTIPLE KIDNEY TRAITS, INCLUDING EGFR AND CKD, IN THIS MULTI-ETHNIC STUDY. EPIGENETIC BIOMARKERS WHICH REFLECT THE SYSTEMIC EFFECTS OF AGE-RELATED MECHANISMS SUCH AS IMMUNOSENESCENCE, INFLAMMAGING AND OXIDATIVE STRESS MAY HAVE IMPORTANT MECHANISTIC OR PROGNOSTIC ROLES IN KIDNEY DISEASE. OUR STUDY HIGHLIGHTS NEW FINDINGS LINKING KIDNEY DISEASE TO BIOLOGICAL AGING, AND OPPORTUNITIES WARRANTING FUTURE INVESTIGATION INTO DNA METHYLATION BIOMARKERS FOR PROGNOSTIC OR RISK STRATIFICATION IN KIDNEY DISEASE.	2021	
                                                                                                                                                                                                                                                                                                                                                                                  
20  1955  34 EPIGENETIC AGE ACCELERATION PREDICTS CANCER, CARDIOVASCULAR, AND ALL-CAUSE MORTALITY IN A GERMAN CASE COHORT. BACKGROUND: PREVIOUS STUDIES HAVE DEVELOPED MODELS PREDICTING METHYLATION AGE FROM DNA METHYLATION IN BLOOD AND OTHER TISSUES (EPIGENETIC CLOCK) AND SUGGESTED THE DIFFERENCE BETWEEN DNA METHYLATION AND CHRONOLOGICAL AGES AS A MARKER OF HEALTHY AGING. THE GOAL OF THIS STUDY WAS TO CONFIRM AND EXPAND SUCH OBSERVATIONS BY INVESTIGATING WHETHER DIFFERENT CONCEPTS OF THE EPIGENETIC CLOCKS IN A POPULATION-BASED COHORT ARE ASSOCIATED WITH CANCER, CARDIOVASCULAR, AND ALL-CAUSE MORTALITY. RESULTS: DNA METHYLATION AGE WAS ESTIMATED IN A COHORT OF 1863 OLDER PEOPLE, AND THE DIFFERENCE BETWEEN AGE PREDICTED BY DNA METHYLATION AND CHRONOLOGICAL AGE (DELTAAGE) WAS CALCULATED. A CASE-COHORT DESIGN AND WEIGHTED PROPORTIONAL COX HAZARD MODELS WERE USED TO ESTIMATE ASSOCIATIONS OF DELTAAGE WITH CANCER, CARDIOVASCULAR, AND ALL-CAUSE MORTALITY. HAZARD RATIOS FOR DELTAAGE (PER 5 YEARS) CALCULATED USING THE EPIGENETIC CLOCK DEVELOPED BY HORVATH WERE 1.23 (95 % CI 1.10-1.38) FOR ALL-CAUSE MORTALITY, 1.22 (95 % CI 1.03-1.45) FOR CANCER MORTALITY, AND 1.19 (95 % CI 0.98-1.43) FOR CARDIOVASCULAR MORTALITY AFTER ADJUSTMENT FOR BATCH EFFECTS, AGE, SEX, EDUCATIONAL LEVEL, HISTORY OF CHRONIC DISEASES, HYPERTENSION, SMOKING STATUS, BODY MASS INDEX, AND LEUCOCYTE DISTRIBUTION. ASSOCIATIONS WERE SIMILAR BUT WEAKER FOR DELTAAGE CALCULATED USING THE EPIGENETIC CLOCK DEVELOPED BY HANNUM. CONCLUSIONS: THESE RESULTS SHOW THAT AGE ACCELERATION IN TERMS OF THE DIFFERENCE BETWEEN AGE PREDICTED BY DNA METHYLATION AND CHRONOLOGICAL AGE IS AN INDEPENDENT PREDICTOR OF ALL-CAUSE AND CAUSE-SPECIFIC MORTALITY AND MAY BE USEFUL AS A GENERAL MARKER OF HEALTHY AGING.	2016