1 4484 170 MOLECULAR SIGNATURE OF CAID SYNDROME: NONCANONICAL ROLES OF SGO1 IN REGULATION OF TGF-BETA SIGNALING AND EPIGENOMICS. BACKGROUND & AIMS: A GENERALIZED HUMAN PACEMAKING SYNDROME, CHRONIC ATRIAL AND INTESTINAL DYSRHYTHMIA (CAID) (OMIM 616201), IS CAUSED BY A HOMOZYGOUS SGO1 MUTATION (K23E), LEADING TO CHRONIC INTESTINAL PSEUDO-OBSTRUCTION AND ARRHYTHMIAS. BECAUSE CAID PATIENTS DO NOT SHOW PHENOTYPES CONSISTENT WITH PERTURBATION OF KNOWN ROLES OF SGO1, WE HYPOTHESIZED THAT NONCANONICAL ROLES OF SGO1 DRIVE THE CLINICAL MANIFESTATIONS OBSERVED. METHODS: TO IDENTIFY A MOLECULAR SIGNATURE FOR CAID SYNDROME, WE ACHIEVED UNBIASED SCREENS IN CELL LINES AND GUT TISSUES FROM CAID PATIENTS VS WILD-TYPE CONTROLS. WE PERFORMED RNA SEQUENCING ALONG WITH STABLE ISOTOPE LABELING WITH AMINO ACIDS IN CELL CULTURE. IN ADDITION, WE DETERMINED THE GENOME-WIDE DNA METHYLATION AND CHROMATIN ACCESSIBILITY SIGNATURES USING REDUCED REPRESENTATIVE BISULFITE SEQUENCING AND ASSAY FOR TRANSPOSASE-ACCESSIBLE CHROMATIN WITH HIGH-THROUGHPUT SEQUENCING. FUNCTIONAL STUDIES INCLUDED PATCH-CLAMP, QUANTITATION OF TRANSFORMING GROWTH FACTOR-BETA (TGF-BETA) SIGNALING, AND IMMUNOHISTOCHEMISTRY IN CAID PATIENT GUT BIOPSY SPECIMENS. RESULTS: PROTEOME AND TRANSCRIPTOME STUDIES CONVERGE ON CELL-CYCLE REGULATION, CARDIAC CONDUCTION, AND SMOOTH MUSCLE REGULATION AS DRIVERS OF CAID SYNDROME. SPECIFICALLY, THE INWARD RECTIFIER CURRENT, AN IMPORTANT REGULATOR OF CELLULAR FUNCTION, WAS DISRUPTED. IMMUNOHISTOCHEMISTRY CONFIRMED OVEREXPRESSION OF BUDDING UNINHIBITED BY BENZIMIDAZOLES 1 (BUB1) IN PATIENTS, IMPLICATING THE TGF-BETA PATHWAY IN CAID PATHOGENESIS. CANONICAL TGF-BETA SIGNALING WAS UP-REGULATED AND UNCOUPLED FROM NONCANONICAL SIGNALING IN CAID PATIENTS. REDUCED REPRESENTATIVE BISULFITE SEQUENCING AND ASSAY FOR TRANSPOSASE-ACCESSIBLE CHROMATIN WITH HIGH-THROUGHPUT SEQUENCING EXPERIMENTS SHOWED SIGNIFICANT CHANGES OF CHROMATIN STATES IN CAID, POINTING TO EPIGENETIC REGULATION AS A POSSIBLE PATHOLOGIC MECHANISM. CONCLUSIONS: OUR FINDINGS POINT TO IMPAIRED INWARD RECTIFIER POTASSIUM CURRENT, DYSREGULATION OF CANONICAL TGF-BETA SIGNALING, AND EPIGENETIC REGULATION AS POTENTIAL DRIVERS OF INTESTINAL AND CARDIAC MANIFESTATIONS OF CAID SYNDROME. TRANSCRIPT PROFILING AND GENOMICS DATA ARE AS FOLLOWS: REPOSITORY URL: HTTPS://WWW.NCBI.NLM.NIH.GOV/GEO; SUPERSERIES GSE110612 WAS COMPOSED OF THE FOLLOWING SUBSERIES: GSE110309, GSE110576, AND GSE110601. 2019 2 746 17 CANNABIS TERATOLOGY EXPLAINS CURRENT PATTERNS OF COLORADAN CONGENITAL DEFECTS: THE CONTRIBUTION OF INCREASED CANNABINOID EXPOSURE TO RISING TERATOLOGICAL TRENDS. RISING DELTA9-TETRAHYDROCANNABINOL CONCENTRATIONS IN MODERN CANNABIS INVITES INVESTIGATION OF THE TERATOLOGICAL IMPLICATIONS OF PRENATAL CANNABIS EXPOSURE. DATA FROM COLORADO RESPONDS TO CHILDREN WITH SPECIAL NEEDS (CRCSN), NATIONAL SURVEY OF DRUG USE AND HEALTH, AND DRUG ENFORCEMENT AGENCY WAS ANALYZED. SEVEN, 40, AND 2 DEFECTS WERE RISING, FLAT, AND FALLING, RESPECTIVELY, AND 10/12 SUMMARY INDICES ROSE. ATRIAL SEPTAL DEFECT, SPINA BIFIDA, MICROCEPHALUS, DOWN'S SYNDROME, VENTRICULAR SEPTAL DEFECT, AND PATENT DUCTUS ARTERIOSUS ROSE, AND ALONG WITH CENTRAL NERVOUS SYSTEM, CARDIOVASCULAR, GENITOURINARY, RESPIRATORY, CHROMOSOMAL, AND MUSCULOSKELETAL DEFECTS ROSE 5 TO 37 TIMES FASTER THAN THE BIRTH RATE (3.3%) TO GENERATE AN EXCESS OF 11 753 (22%) MAJOR ANOMALIES. CANNABIS WAS THE ONLY DRUG WHOSE USE GREW FROM 2000 TO 2014 WHILE PAIN RELIEVERS, COCAINE, ALCOHOL, AND TOBACCO DID NOT. THE CORRELATION OF CANNABIS USE WITH MAJOR DEFECTS IN 2014 (2019 DATASET) WAS R = .77, P = .0011. MULTIPLE CANNABINOIDS WERE LINKED WITH SUMMARY MEASURES OF CONGENITAL ANOMALIES AND WERE ROBUST TO MULTIVARIATE ADJUSTMENT. 2019 3 781 37 CELL-FREE FILTRATES (CFF) AS VECTORS OF A TRANSMISSIBLE PATHOLOGIC TISSUE MEMORY CODE: A HYPOTHETICAL AND NARRATIVE REVIEW. CELLULAR MEMORY IS A CONTROVERSIAL CONCEPT REPRESENTING THE ABILITY OF CELLS TO "WRITE AND MEMORIZE" STRESSFUL EXPERIENCES VIA EPIGENETIC OPERATORS. THE PROGRESSIVE COURSE OF CHRONIC, NON-COMMUNICABLE DISEASES SUCH AS TYPE 2 DIABETES MELLITUS, CANCER, AND ARTERIOSCLEROSIS, IS LIKELY DRIVEN THROUGH AN ABNORMAL EPIGENETIC REPROGRAMMING, FOSTERING THE HYPOTHESIS OF A CELLULAR PATHOLOGIC MEMORY. ACCORDINGLY, CULTURED DIABETIC AND CANCER PATIENT-DERIVED CELLS RECALL BEHAVIORAL TRAITS AS WHEN IN THE DONOR'S ORGANISM IRRESPECTIVE TO CULTURE TIME AND CONDITIONS. HERE, WE ANALYZE THE DATA OF STUDIES CONDUCTED BY OUR GROUP AND LED BY A CASCADE OF HYPOTHESIS, IN WHICH WE AIMED TO VALIDATE THE HYPOTHETICAL EXISTENCE AND TRANSMISSIBILITY OF A CELLULAR PATHOLOGIC MEMORY IN DIABETES, ARTERIOSCLEROTIC PERIPHERAL ARTERIAL DISEASE, AND CANCER. THESE EXPERIMENTS WERE BASED ON THE ADMINISTRATION TO OTHERWISE HEALTHY ANIMALS OF CELL-FREE FILTRATES PREPARED FROM HUMAN PATHOLOGIC TISSUE SAMPLES REPRESENTATIVE OF EACH DISEASE CONDITION. THE ADMINISTRATION OF EACH PATHOLOGIC TISSUE HOMOGENATE CONSISTENTLY INDUCED THE FAITHFUL RECAPITULATION OF: (1) DIABETIC ARCHETYPICAL CHANGES IN CUTANEOUS ARTERIOLES AND NERVES. (2) NON-THROMBOTIC ARTERIOSCLEROTIC THICKENING, COLLAGENOUS ARTERIAL ENCROACHMENT, ABERRANT ANGIOGENESIS, AND VASCULAR REMODELING. (3) PRE-MALIGNANT AND MALIGNANT EPITHELIAL AND MESENCHYMAL TUMORS IN DIFFERENT ORGANS; ALL EVOCATIVE OF THE DONOR'S TISSUE HISTOPATHOLOGY AND WITH NO BARRIERS FOR INTERSPECIES TRANSMISSION. WE HYPOTHESIZE THAT HOMOGENATES CONTAIN PATHOLOGIC TISSUE MEMORY CODES REPRESENTED IN SOLUBLE DRIVERS THAT "INFILTRATE" HOST'S ANIMAL CELLS, AND ULTIMATELY IMPOSE THEIR PHENOTYPIC SIGNATURES. THE IDENTIFICATION AND VALIDATION OF THE ACTORS IN BEHIND MAY PAVE THE WAY FOR FUTURE THERAPIES. 2022 4 6569 24 TRANSPLANTATION OF EPIGENETICALLY MODIFIED ADULT CARDIAC C-KIT+ CELLS RETARDS REMODELING AND IMPROVES CARDIAC FUNCTION IN ISCHEMIC HEART FAILURE MODEL. CARDIAC C-KIT+ CELLS HAVE A MODEST CARDIOGENIC POTENTIAL THAT COULD LIMIT THEIR EFFICACY IN HEART DISEASE TREATMENT. THE PRESENT STUDY WAS DESIGNED TO AUGMENT THE CARDIOGENIC POTENTIAL OF CARDIAC C-KIT+ CELLS THROUGH CLASS I HISTONE DEACETYLASE (HDAC) INHIBITION AND EVALUATE THEIR THERAPEUTIC POTENCY IN THE CHRONIC HEART FAILURE (CHF) ANIMAL MODEL. MYOCARDIAL INFARCTION (MI) WAS CREATED BY CORONARY ARTERY OCCLUSION IN RATS. C-KIT+ CELLS WERE TREATED WITH MOCETINOSTAT (MOCE), A SPECIFIC CLASS I HDAC INHIBITOR. AT 3 WEEKS AFTER MI, CHF ANIMALS WERE RETROGRADELY INFUSED WITH UNTREATED (CONTROL) OR MOCE-TREATED C-KIT+ CELLS (MOCE/C-KIT+ CELLS) AND EVALUATED AT 3 WEEKS AFTER CELL INFUSION. WE FOUND THAT CLASS I HDAC INHIBITION IN C-KIT+ CELLS ELEVATED THE LEVEL OF ACETYLATED HISTONE H3 (ACH3) AND INCREASED ACH3 LEVELS IN THE PROMOTER REGIONS OF PLURIPOTENT AND CARDIAC-SPECIFIC GENES. EPIGENETIC CHANGES WERE ACCOMPANIED BY INCREASED EXPRESSION OF CARDIAC-SPECIFIC MARKERS. TRANSPLANTATION OF CHF RATS WITH EITHER CONTROL OR MOCE/C-KIT+ CELLS RESULTED IN AN IMPROVEMENT IN CARDIAC FUNCTION, RETARDATION OF CHF REMODELING MADE EVIDENT BY INCREASED VASCULARIZATION AND SCAR SIZE, AND CARDIOMYOCYTE HYPERTROPHY REDUCTION. COMPARED WITH CHF INFUSED WITH CONTROL CELLS, INFUSION OF MOCE/C-KIT+ CELLS RESULTED IN A FURTHER REDUCTION IN LEFT VENTRICLE END-DIASTOLIC PRESSURE AND TOTAL COLLAGEN AND AN INCREASE IN INTERLEUKIN-6 EXPRESSION. THE LOW ENGRAFTMENT OF INFUSED CELLS SUGGESTS THAT PARACRINE EFFECTS MIGHT ACCOUNT FOR THE BENEFICIAL EFFECTS OF C-KIT+ CELLS IN CHF. IN CONCLUSION, SELECTIVE INHIBITION OF CLASS I HDACS INDUCED EXPRESSION OF CARDIAC MARKERS IN C-KIT+ CELLS AND PARTIALLY AUGMENTED THE EFFICACY OF THESE CELLS FOR CHF REPAIR. SIGNIFICANCE: THE STUDY HAS SHOWN THAT SELECTIVE CLASS 1 HISTONE DEACETYLASE INHIBITION IS SUFFICIENT TO REDIRECT C-KIT+ CELLS TOWARD A CARDIAC FATE. EPIGENETICALLY MODIFIED C-KIT+ CELLS IMPROVED CONTRACTILE FUNCTION AND RETARDED REMODELING OF THE CONGESTIVE HEART FAILURE HEART. THIS STUDY PROVIDES NEW INSIGHTS INTO THE EFFICACY OF CARDIAC C-KIT+ CELLS IN THE ISCHEMIC HEART FAILURE MODEL. 2015 5 5735 26 SMALL VESSEL DISEASE-RELATED DEMENTIA: AN INVALID NEUROVASCULAR COUPLING? THE ARTERIOSCLEROSIS-DEPENDENT ALTERATION OF BRAIN PERFUSION IS ONE OF THE MAJOR DETERMINANTS IN SMALL VESSEL DISEASE, SINCE SMALL VESSELS HAVE A PIVOTAL ROLE IN THE BRAIN'S AUTOREGULATION. NEVERTHELESS, AS FAR AS WE KNOW, ENDOTHELIUM DISTRESS CAN POTENTIATE THE FLOW DYSREGULATION AND LEAD TO SUBCORTICAL VASCULAR DEMENTIA THAT IS RELATED TO SMALL VESSEL DISEASE (SVD), ALSO BEING DEFINED AS SUBCORTICAL VASCULAR DEMENTIA (SVAD), AS WELL AS MICROGLIA ACTIVATION, CHRONIC HYPOXIA AND HYPOPERFUSION, VESSEL-TONE DYSREGULATION, ALTERED ASTROCYTES, AND PERICYTES FUNCTIONING BLOOD-BRAIN BARRIER DISRUPTION. THE MOLECULAR BASIS OF THIS PATHOLOGY REMAINS CONTROVERSIAL. THE APPARENT CONSEQUENCE (OR A FIRST EVENT, TOO) IS THE MACROSCOPIC ALTERATION OF THE NEUROVASCULAR COUPLING. HERE, WE EXAMINED THE POSSIBLE MECHANISMS THAT LEAD A HEALTHY AGING PROCESS TOWARDS SUBCORTICAL DEMENTIA. WE REMARKED THAT SVD AND WHITE MATTER ABNORMALITIES RELATED TO AGE COULD BE ACCELERATED AND POTENTIATED BY DIFFERENT VASCULAR RISK FACTORS. VASCULAR FUNCTION CHANGES CAN BE HEAVILY INFLUENCED BY GENETIC AND EPIGENETIC FACTORS, WHICH ARE, TO THE BEST OF OUR KNOWLEDGE, MOSTLY UNKNOWN. METABOLIC DEMANDS, ACTIVE NEUROVASCULAR COUPLING, CORRECT GLYMPHATIC PROCESS, AND ADEQUATE OXIDATIVE AND INFLAMMATORY RESPONSES COULD BE BULWARKS IN DEFENSE OF THE CORRECT AGING PROCESS; THEIR IMPAIRMENTS LEAD TO A POTENTIALLY CATASTROPHIC AND NON-REVERSIBLE CONDITION. 2020 6 1709 30 DYSFUNCTIONAL ERG SIGNALING DRIVES PULMONARY VASCULAR AGING AND PERSISTENT FIBROSIS. VASCULAR DYSFUNCTION IS A HALLMARK OF CHRONIC DISEASES IN ELDERLY. THE CONTRIBUTION OF THE VASCULATURE TO LUNG REPAIR AND FIBROSIS IS NOT FULLY UNDERSTOOD. HERE, WE PERFORMED AN EPIGENETIC AND TRANSCRIPTIONAL ANALYSIS OF LUNG ENDOTHELIAL CELLS (ECS) FROM YOUNG AND AGED MICE DURING THE RESOLUTION OR PROGRESSION OF BLEOMYCIN-INDUCED LUNG FIBROSIS. WE IDENTIFIED THE TRANSCRIPTION FACTOR ETS-RELATED GENE (ERG) AS PUTATIVE ORCHESTRATOR OF LUNG CAPILLARY HOMEOSTASIS AND REPAIR, AND WHOSE FUNCTION IS DYSREGULATED IN AGING. ERG DYSREGULATION IS ASSOCIATED WITH REDUCED CHROMATIN ACCESSIBILITY AND MALADAPTIVE TRANSCRIPTIONAL RESPONSES TO INJURY. LOSS OF ENDOTHELIAL ERG ENHANCES PARACRINE FIBROBLAST ACTIVATION IN VITRO, AND IMPAIRS LUNG FIBROSIS RESOLUTION IN YOUNG MICE IN VIVO. SCRNA-SEQ OF ERG DEFICIENT MOUSE LUNGS REVEALES TRANSCRIPTIONAL AND FIBROGENIC ABNORMALITIES RESEMBLING THOSE ASSOCIATED WITH AGING AND HUMAN LUNG FIBROSIS, INCLUDING REDUCED NUMBER OF GENERAL CAPILLARY (GCAP) ECS. OUR FINDINGS DEMONSTRATE THAT LUNG ENDOTHELIAL CHROMATIN REMODELING DETERIORATES WITH AGING LEADING TO ABNORMAL TRANSCRIPTION, VASCULAR DYSREPAIR, AND PERSISTENT FIBROSIS FOLLOWING INJURY. 2022 7 5826 23 STRESS SIGNAL NETWORK BETWEEN HYPOXIA AND ER STRESS IN CHRONIC KIDNEY DISEASE. CHRONIC KIDNEY DISEASE (CKD) IS CHARACTERIZED BY AN IRREVERSIBLE DECREASE IN KIDNEY FUNCTION AND INDUCTION OF VARIOUS METABOLIC DYSFUNCTIONS. ACCUMULATED FINDINGS REVEAL THAT CHRONIC HYPOXIC STRESS AND ENDOPLASMIC RETICULUM (ER) STRESS ARE INVOLVED IN A RANGE OF PATHOGENIC CONDITIONS, INCLUDING THE PROGRESSION OF CKD. BECAUSE OF THE PRESENCE OF AN ARTERIOVENOUS OXYGEN SHUNT, THE KIDNEY IS THOUGHT TO BE SUSCEPTIBLE TO HYPOXIA. CHRONIC KIDNEY HYPOXIA IS INDUCED BY A NUMBER OF PATHOGENIC CONDITIONS, INCLUDING RENAL ISCHEMIA, REDUCED PERITUBULAR CAPILLARY, AND TUBULOINTERSTITIAL FIBROSIS. THE ER IS AN ORGANELLE WHICH HELPS MAINTAIN THE QUALITY OF PROTEINS THROUGH THE UNFOLDED PROTEIN RESPONSE (UPR) PATHWAY, AND ER DYSFUNCTION ASSOCIATED WITH MALADAPTIVE UPR ACTIVATION IS NAMED ER STRESS. ER STRESS IS REPORTED TO BE RELATED TO SOME OF THE EFFECTS OF PATHOGENESIS IN KIDNEY, PARTICULARLY IN THE PODOCYTE SLIT DIAPHRAGM AND TUBULOINTERSTITIUM. FURTHERMORE, CHRONIC HYPOXIA MEDIATES ER STRESS IN BLOOD VESSEL ENDOTHELIAL CELLS AND TUBULOINTERSTITIUM VIA SEVERAL MECHANISMS, INCLUDING OXIDATIVE STRESS, EPIGENETIC ALTERATION, LIPID METABOLISM, AND THE AKT PATHWAY. IN SUMMARY, A GROWING CONSENSUS CONSIDERS THAT THESE STRESSES INTERACT VIA COMPLICATED STRESS SIGNAL NETWORKS, WHICH LEADS TO THE EXACERBATION OF CKD (FIGURE 1). THIS STRESS SIGNAL NETWORK MIGHT BE A TARGET FOR INTERVENTIONS AIMED AT AMELIORATING CKD. 2017 8 487 34 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 9 444 32 AORTIC AND CAROTID ARTERIAL STIFFNESS AND EPIGENETIC REGULATOR GENE EXPRESSION CHANGES PRECEDE BLOOD PRESSURE RISE IN STROKE-PRONE DAHL SALT-SENSITIVE HYPERTENSIVE RATS. MULTIPLE CLINICAL STUDIES SHOW THAT ARTERIAL STIFFNESS, MEASURED AS PULSE WAVE VELOCITY (PWV), PRECEDES HYPERTENSION AND IS AN INDEPENDENT PREDICTOR OF HYPERTENSION END ORGAN DISEASES INCLUDING STROKE, CARDIOVASCULAR DISEASE AND CHRONIC KIDNEY DISEASE. RISK FACTOR STUDIES FOR ARTERIAL STIFFNESS IMPLICATE AGE, HYPERTENSION AND SODIUM. HOWEVER, CAUSAL MECHANISMS LINKING RISK FACTOR TO ARTERIAL STIFFNESS REMAIN TO BE ELUCIDATED. HERE, WE STUDIED THE CAUSAL RELATIONSHIP OF ARTERIAL STIFFNESS AND HYPERTENSION IN THE NA-INDUCED, STROKE-PRONE DAHL SALT-SENSITIVE (S) HYPERTENSIVE RAT MODEL, AND ANALYZED PUTATIVE MOLECULAR MECHANISMS. STROKE-PRONE AND NON-STROKE-PRONE MALE AND FEMALE RATS WERE STUDIED AT 3- AND 6-WEEKS OF AGE FOR ARTERIAL STIFFNESS (PWV, STRAIN), BLOOD PRESSURE, VESSEL WALL HISTOLOGY, AND GENE EXPRESSION CHANGES. STUDIES SHOWED THAT INCREASED LEFT CAROTID AND AORTIC ARTERIAL STIFFNESS PRECEDED HYPERTENSION, PULSE PRESSURE WIDENING, AND STRUCTURAL WALL CHANGES AT THE 6-WEEK TIME-POINT. INSTEAD, DIFFERENTIAL GENE INDUCTION WAS DETECTED IMPLICATING MOLECULAR-FUNCTIONAL CHANGES IN EXTRACELLULAR MATRIX (ECM) STRUCTURAL CONSTITUENTS, MODIFIERS, CELL ADHESION, AND MATRICELLULAR PROTEINS, AS WELL AS IN ENDOTHELIAL FUNCTION, APOPTOSIS BALANCE, AND EPIGENETIC REGULATORS. IMMUNOSTAINING TESTING HISTONE MODIFIERS EP300, HDAC3, AND PRMT5 LEVELS CONFIRMED CAROTID ARTERY-UPREGULATION IN ALL THREE LAYERS: ENDOTHELIAL, SMOOTH MUSCLE AND ADVENTITIAL CELLS. OUR STUDY RECAPITULATES OBSERVATIONS IN HUMANS THAT GIVEN SALT-SENSITIVITY, INCREASED NA-INTAKE INDUCED ARTERIAL STIFFNESS BEFORE HYPERTENSION, INCREASED PULSE PRESSURE, AND STRUCTURAL VESSEL WALL CHANGES. DIFFERENTIAL GENE EXPRESSION CHANGES ASSOCIATED WITH ARTERIAL STIFFNESS SUGGEST A MOLECULAR MECHANISM LINKING SODIUM TO FULL-VESSEL WALL RESPONSE AFFECTING GENE-NETWORKS INVOLVED IN VASCULAR ECM STRUCTURE-FUNCTION, APOPTOSIS BALANCE, AND EPIGENETIC REGULATION. 2014 10 2378 29 EPIGENETIC REGULATION OF VASCULAR SMOOTH MUSCLE CELL PHENOTYPE SWITCHING IN ATHEROSCLEROTIC ARTERY REMODELING: A MINI-REVIEW. ATHEROSCLEROSIS IS A CHRONIC INFLAMMATORY DISEASE CHARACTERIZED BY EXTENSIVE REMODELING OF MEDIUM AND LARGE-SIZED ARTERIES. INWARD REMODELING (=LUMEN SHRINKAGE) OF THE VASCULAR WALLS IS THE UNDERLYING CAUSE FOR ISCHEMIA IN TARGET ORGANS. THEREFORE, INWARD REMODELING CAN BE CONSIDERED THE PREDOMINANT FEATURE OF ATHEROSCLEROTIC PATHOLOGY. OUTWARD REMODELING (=LUMEN ENLARGEMENT) IS A PHYSIOLOGICAL RESPONSE COMPENSATING FOR LUMEN SHRINKAGE CAUSED BY NEOINTIMAL HYPERPLASIA, BUT AS A PATHOLOGICAL RESPONSE TO CHANGES IN BLOOD FLOW, OUTWARD REMODELING LEADS TO SUBSTANTIAL ARTERIAL WALL THINNING. THINNED VASCULAR WALLS ARE PRONE TO RUPTURE, AND SUBSEQUENT THROMBUS FORMATION ACCOUNTS FOR THE MAJORITY OF ACUTE CARDIOVASCULAR EVENTS. PATHOLOGICAL REMODELING IS DRIVEN BY INFLAMMATORY CELLS WHICH INDUCE VASCULAR SMOOTH MUSCLE CELLS TO SWITCH FROM QUIESCENT TO A PROLIFERATIVE AND MIGRATORY PHENOTYPE. AFTER DECADES OF INTENSIVE RESEARCH, THE MOLECULAR MECHANISMS OF ARTERIAL REMODELING ARE STARTING TO UNFOLD. IN THIS MINI-REVIEW, WE SUMMARIZE THE CURRENT KNOWLEDGE OF THE EPIGENETIC AND TRANSCRIPTIONAL REGULATION OF VASCULAR SMOOTH MUSCLE CELL PHENOTYPE SWITCHING FROM THE CONTRACTILE TO THE SYNTHETIC PHENOTYPE INVOLVED IN ARTERIAL REMODELING AND DISCUSS POTENTIAL THERAPEUTIC OPTIONS. 2021 11 274 31 AGE-RELATED CHANGES IN DNA METHYLATION AFFECT RENAL HISTOLOGY AND POST-TRANSPLANT FIBROSIS. DURING AGEING, KIDNEY FUNCTION DECREASES DUE TO RENAL TUBULAR ATROPHY, INTERSTITIAL FIBROSIS, GLOMERULOSCLEROSIS AND ARTERIOSCLEROSIS. RECENTLY, CHANGES IN DNA METHYLATION WERE SHOWN TO CONTRIBUTE TO VARIOUS AGEING PROCESSES. HOWEVER, IT IS UNKNOWN WHETHER SUCH CHANGES ALSO CONTRIBUTE TO AGE-RELATED KIDNEY DYSFUNCTION. TO ASSESS THIS, WE PROFILED GENOME-WIDE CHANGES IN DNA METHYLATION (OVER 800 000 CPG SITES) IN 95 RENAL BIOPSIES OBTAINED PRIOR TO KIDNEY TRANSPLANTATION FROM DONORS AGED 16 TO 73 YEARS. DONOR AGE SIGNIFICANTLY ASSOCIATED WITH THE METHYLATION OF 92 778 CPGS (FALSE DISCOVERY RATE UNDER 0.05), CORRESPONDING TO 10 285 DIFFERENTIALLY METHYLATED REGIONS. THESE REGIONS WERE MOST FREQUENTLY LOCATED IN GENES INVOLVED IN THE WNT/BETA-CATENIN SIGNALING PATHWAY. USING AN INDEPENDENT COHORT OF 67 BIOPSIES, WE AUTONOMOUSLY VALIDATED THESE FINDINGS. INTERESTINGLY, THE METHYLATION STATUS OF THESE 92 778 AGE-RELATED CPGS WAS ASSOCIATED WITH GLOMERULOSCLEROSIS (34.4% OF CPGS AT A FALSE DISCOVERY RATE UNDER 0.05) AND INTERSTITIAL FIBROSIS (0.9%) AND GRAFT FUNCTION AT ONE YEAR AFTER TRANSPLANTATION, BUT NOT WITH TUBULAR ATROPHY AND ARTERIOSCLEROSIS. NO ASSOCIATION WAS OBSERVED WITH ANY OF THESE PATHOLOGIES AT THE TIME OF TRANSPLANTATION (0% AT A FALSE DISCOVERY RATE UNDER 0.05). THUS, AGE-ASSOCIATED CHANGES IN DNA METHYLATION AT THE TIME OF TRANSPLANTATION PREDICT FUTURE INJURY OF TRANSPLANTED KIDNEYS. SPECIFICALLY, OUR EPIGENOME-WIDE ASSOCIATION STUDY DEMONSTRATES THAT EPIGENETIC RENAL AGEING IS IMPLICATED IN PROGRESSIVE FIBROSIS IN BOTH THE GLOMERULUS AND THE INTERSTITIUM. 2019 12 3064 47 GENOME-WIDE DNA METHYLATION ENCODES CARDIAC TRANSCRIPTIONAL REPROGRAMMING IN HUMAN ISCHEMIC HEART FAILURE. ISCHEMIC CARDIOMYOPATHY (ICM) IS THE CLINICAL ENDPOINT OF CORONARY HEART DISEASE AND A LEADING CAUSE OF HEART FAILURE. DESPITE GROWING DEMANDS TO DEVELOP PERSONALIZED APPROACHES TO TREAT ICM, PROGRESS IS LIMITED BY INADEQUATE KNOWLEDGE OF ITS PATHOGENESIS. SINCE EPIGENETICS HAS BEEN IMPLICATED IN THE DEVELOPMENT OF OTHER CHRONIC DISEASES, THE CURRENT STUDY WAS DESIGNED TO DETERMINE WHETHER TRANSCRIPTIONAL AND/OR EPIGENETIC CHANGES ARE SUFFICIENT TO DISTINGUISH ICM FROM OTHER ETIOLOGIES OF HEART FAILURE. SPECIFICALLY, WE HYPOTHESIZE THAT GENOME-WIDE DNA METHYLATION ENCODES TRANSCRIPTIONAL REPROGRAMMING IN ICM. RNA-SEQUENCING ANALYSIS WAS PERFORMED ON HUMAN ISCHEMIC LEFT VENTRICULAR TISSUE OBTAINED FROM PATIENTS WITH END-STAGE HEART FAILURE, WHICH ENRICHED KNOWN TARGETS OF THE POLYCOMB METHYLTRANSFERASE EZH2 COMPARED TO NON-ISCHEMIC HEARTS. COMBINED RNA SEQUENCING AND GENOME-WIDE DNA METHYLATION ANALYSIS REVEALED A ROBUST GENE EXPRESSION PATTERN CONSISTENT WITH SUPPRESSION OF OXIDATIVE METABOLISM, INDUCED ANAEROBIC GLYCOLYSIS, AND ALTERED CELLULAR REMODELING. LASTLY, KLF15 WAS IDENTIFIED AS A PUTATIVE UPSTREAM REGULATOR OF METABOLIC GENE EXPRESSION THAT WAS ITSELF REGULATED BY EZH2 IN A SET DOMAIN-DEPENDENT MANNER. OUR OBSERVATIONS THEREFORE DEFINE A NOVEL ROLE OF DNA METHYLATION IN THE METABOLIC REPROGRAMMING OF ICM. FURTHERMORE, WE IDENTIFY EZH2 AS AN EPIGENETIC REGULATOR OF KLF15 ALONG WITH DNA HYPERMETHYLATION, AND WE PROPOSE A NOVEL MECHANISM THROUGH WHICH CORONARY HEART DISEASE REPROGRAMS THE EXPRESSION OF BOTH INTERMEDIATE ENZYMES AND UPSTREAM REGULATORS OF CARDIAC METABOLISM SUCH AS KLF15. 2019 13 4415 28 MOLECULAR AND CELLULAR MECHANISMS THAT INDUCE ARTERIAL CALCIFICATION BY INDOXYL SULFATE AND P-CRESYL SULFATE. THE PROTEIN-BOUND UREMIC TOXINS, INDOXYL SULFATE (IS) AND P-CRESYL SULFATE (PCS), ARE CONSIDERED TO BE HARMFUL VASCULAR TOXINS. ARTERIAL MEDIA CALCIFICATION, OR THE DEPOSITION OF CALCIUM PHOSPHATE CRYSTALS IN THE ARTERIES, CONTRIBUTES SIGNIFICANTLY TO CARDIOVASCULAR COMPLICATIONS, INCLUDING LEFT VENTRICULAR HYPERTROPHY, HYPERTENSION, AND IMPAIRED CORONARY PERFUSION IN THE ELDERLY AND PATIENTS WITH CHRONIC KIDNEY DISEASE (CKD) AND DIABETES. RECENTLY, WE REPORTED THAT BOTH IS AND PCS TRIGGER MODERATE TO SEVERE CALCIFICATION IN THE AORTA AND PERIPHERAL VESSELS OF CKD RATS. THIS REVIEW DESCRIBES THE MOLECULAR AND CELLULAR MECHANISMS BY WHICH THESE UREMIC TOXINS INDUCE ARTERIAL MEDIA CALCIFICATION. A COMPLEX INTERPLAY BETWEEN INFLAMMATION, COAGULATION, AND LIPID METABOLISM PATHWAYS, INFLUENCED BY EPIGENETIC FACTORS, IS CRUCIAL IN IS/PCS-INDUCED ARTERIAL MEDIA CALCIFICATION. HIGH LEVELS OF GLUCOSE ARE LINKED TO THESE EVENTS, SUGGESTING THAT A GOOD BALANCE BETWEEN GLUCOSE AND LIPID LEVELS MIGHT BE IMPORTANT. ON THE CELLULAR LEVEL, EFFECTS ON ENDOTHELIAL CELLS, WHICH ACT AS THE PRIMARY SENSORS OF CIRCULATING PATHOLOGICAL TRIGGERS, MIGHT BE AS IMPORTANT AS THOSE ON VASCULAR SMOOTH MUSCLE CELLS. ENDOTHELIAL DYSFUNCTION, PROVOKED BY IS AND PCS TRIGGERED OXIDATIVE STRESS, MAY BE CONSIDERED A KEY EVENT IN THE ONSET AND DEVELOPMENT OF ARTERIAL MEDIA CALCIFICATION. IN THIS REVIEW A NUMBER OF IMPORTANT OUTSTANDING QUESTIONS SUCH AS THE ROLE OF MIRNA'S, PHENOTYPIC SWITCHING OF BOTH ENDOTHELIAL AND VASCULAR SMOOTH MUSCLE CELLS AND NEW TYPES OF PROGRAMMED CELL DEATH IN ARTERIAL MEDIA CALCIFICATION RELATED TO PROTEIN-BOUND UREMIC TOXINS ARE PUT FORWARD AND DISCUSSED. 2020 14 4170 23 MEETING REPORT: ISN FOREFRONTS IN NEPHROLOGY ON ENDOTHELIAL BIOLOGY AND RENAL DISEASE: FROM BENCH TO PREVENTION. THIS ISN-SPONSORED FOREFRONT IN NEPHROLOGY MEETING, WHICH HAS BROUGHT TOGETHER 120 SCIENTISTS FROM 21 COUNTRIES, HAS BEEN CONCERNED WITH VARIOUS ASPECTS OF ENDOTHELIAL FUNCTION AND DYSFUNCTION AND THEIR CONTRIBUTION TO PROGRESSION OF CHRONIC KIDNEY DISEASE AND/OR ITS CARDIOVASCULAR COMPLICATIONS. THE FOLLOWING THEMES WERE DISCUSSED IN GREAT DEPTH: (1) PHENOTYPICAL CHANGES IN THE VASCULAR ENDOTHELIUM - PERMEABILITY, SENESCENCE, AND APOPTOSIS; (2) REGULATION OF ENDOTHELIAL NITRIC OXIDE (NO) SYNTHASE FUNCTION - CAVEOLAR AND SHEAR STRESS MECHANISMS, EPIGENETIC REGULATION, S-NITROSYLATION, AND RHO-KINASE REGULATION; (3) OXIDATIVE STRESS AND HYPOXIA-INDUCED CHANGES; (4) ORGANELLAR DYSFUNCTION - LYSOSOMES, MITOCHONDRIA, AND ENDOPLASMIC RETICULUM; (5) NO-INDEPENDENT MECHANISMS OF VASOMOTION - EPOXIDES, HEME OXYGENASE-1 AND CARBON MONOXIDE, THROMBOXANE, TUMOR NECROSIS FACTOR-ALPHA, AND URIC ACID; (6) ENDOTHELIAL CROSSTALK WITH PODOCYTES, MONOCYTES, SMOOTH MUSCLE CELLS, AND PLATELETS; (7) CANDIDATE CLINICAL BIOMARKERS OF ENDOTHELIAL DYSFUNCTION - FUNCTIONAL TESTING OF MACRO- AND MICRO-VASCULAR FUNCTIONS, SURROGATE MARKERS, CIRCULATING DETACHED ENDOTHELIAL CELLS, AND ENDOTHELIAL PRECURSOR CELLS; AND CULMINATED IN ROUND TABLE DISCUSSION ON THE DIAGNOSIS OF ENDOTHELIAL DYSFUNCTION AND ITS TREATMENT OPTIONS. IN CONCLUSION, THIS MEETING HAS FOCUSED ON SEVERAL KEY PROBLEMS OF ENDOTHELIAL CELL PATHOBIOLOGY RELEVANT TO CHRONIC KIDNEY DISEASE. 2006 15 2385 31 EPIGENETIC REGULATORS OF THE REVASCULARIZATION RESPONSE TO CHRONIC ARTERIAL OCCLUSION. PERIPHERAL ARTERIAL DISEASE (PAD) IS THE LEADING CAUSE OF LOWER LIMB AMPUTATION AND ESTIMATED TO AFFECT OVER 202 MILLION PEOPLE WORLDWIDE. PAD IS CAUSED BY ATHEROSCLEROTIC LESIONS THAT OCCLUDE LARGE ARTERIES IN THE LOWER LIMBS, LEADING TO INSUFFICIENT BLOOD PERFUSION OF DISTAL TISSUES. GIVEN THE SEVERITY OF THIS CLINICAL PROBLEM, THERE HAS BEEN LONG-STANDING INTEREST IN BOTH UNDERSTANDING HOW CHRONIC ARTERIAL OCCLUSIONS AFFECT MUSCLE TISSUE AND VASCULATURE AND IDENTIFYING THERAPEUTIC APPROACHES CAPABLE OF RESTORING TISSUE COMPOSITION AND VASCULAR FUNCTION TO A HEALTHY STATE. TO DATE, THE MOST WIDELY UTILIZED ANIMAL MODEL FOR PERFORMING SUCH STUDIES HAS BEEN THE ISCHAEMIC MOUSE HINDLIMB. DESPITE NOT BEING A MODEL OF PAD PER SE, THE ISCHAEMIC HINDLIMB MODEL DOES RECAPITULATE SEVERAL KEY ASPECTS OF PAD. FURTHER, IT HAS SERVED AS A VALUABLE PLATFORM UPON WHICH WE HAVE BUILT MUCH OF OUR UNDERSTANDING OF HOW CHRONIC ARTERIAL OCCLUSIONS AFFECT MUSCLE TISSUE COMPOSITION, MUSCLE REGENERATION AND ANGIOGENESIS, AND COLLATERAL ARTERIOGENESIS. RECENTLY, THERE HAS BEEN A GLOBAL SURGE IN RESEARCH AIMED AT UNDERSTANDING HOW GENE EXPRESSION IS REGULATED BY EPIGENETIC FACTORS (I.E. NON-CODING RNAS, HISTONE POST-TRANSLATIONAL MODIFICATIONS, AND DNA METHYLATION). THUS, PERHAPS NOT UNEXPECTEDLY, MANY RECENT STUDIES HAVE IDENTIFIED ESSENTIAL ROLES FOR EPIGENETIC FACTORS IN REGULATING KEY RESPONSES TO CHRONIC ARTERIAL OCCLUSION(S). IN THIS REVIEW, WE SUMMARIZE THE MECHANISMS OF ACTION OF THESE EPIGENETIC REGULATORS AND HIGHLIGHT SEVERAL RECENT STUDIES INVESTIGATING THE ROLE OF SAID REGULATORS IN THE CONTEXT OF HINDLIMB ISCHAEMIA. IN ADDITION, WE FOCUS ON HOW THESE RECENT ADVANCES IN OUR UNDERSTANDING OF THE ROLE OF EPIGENETICS IN REGULATING RESPONSES TO CHRONIC ARTERIAL OCCLUSION(S) CAN INFORM FUTURE THERAPEUTIC APPLICATIONS TO PROMOTE REVASCULARIZATION AND PERFUSION RECOVERY IN THE SETTING OF PAD. 2019 16 3624 40 IN VIVO FUNCTION OF FLOW-RESPONSIVE CIS-DNA ELEMENTS OF ENOS GENE: A ROLE FOR CHROMATIN-BASED MECHANISMS. BACKGROUND: ENOS (ENDOTHELIAL NITRIC OXIDE SYNTHASE) IS AN ENDOTHELIAL CELL (EC)-SPECIFIC GENE PREDOMINANTLY EXPRESSED IN MEDIUM- TO LARGE-SIZED ARTERIES WHERE ECS EXPERIENCE ATHEROPROTECTIVE LAMINAR FLOW WITH HIGH SHEAR STRESS. DISTURBED FLOW WITH LOWER AVERAGE SHEAR STRESS DECREASES ENOS TRANSCRIPTION, WHICH LEADS TO THE DEVELOPMENT OF ATHEROSCLEROSIS, ESPECIALLY AT BIFURCATIONS AND CURVATURES OF ARTERIES. THIS PROTOTYPIC ARTERIAL EC GENE CONTAINS 2 DISTINCT FLOW-RESPONSIVE CIS-DNA ELEMENTS IN THE PROMOTER, THE SHEAR STRESS RESPONSE ELEMENT (SSRE) AND THE KLF (KRUPPEL-LIKE FACTOR) ELEMENT. PREVIOUS IN VITRO STUDIES SUGGESTED THEIR POSITIVE REGULATORY FUNCTIONS ON FLOW-INDUCED TRANSCRIPTION OF EC GENES INCLUDING ENOS. HOWEVER, THE IN VIVO FUNCTION OF THESE CIS-DNA ELEMENTS REMAINS UNKNOWN. METHODS: INSERTIONAL TRANSGENIC MICE WITH A MUTATION AT EACH FLOW-RESPONSIVE CIS-DNA ELEMENT WERE GENERATED USING A MURINE ENOS PROMOTER-BETA-GALACTOSIDASE REPORTER BY LINKER-SCANNING MUTAGENESIS AND COMPARED WITH EPISOMAL-BASED MUTATIONS IN VITRO. DNA METHYLATION AT THE ENOS PROXIMAL PROMOTER IN MOUSE ECS WAS ASSESSED BY BISULFITE SEQUENCING OR PYROSEQUENCING. RESULTS: WILD TYPE MICE WITH A FUNCTIONAL ENOS PROMOTER-REPORTER TRANSGENE EXHIBITED REDUCED ENDOTHELIAL REPORTER EXPRESSION IN THE ATHEROPRONE REGIONS OF DISTURBED FLOW (N=5). IT IS SURPRISING THAT THE SSRE MUTATION ABROGATED REPORTER EXPRESSION IN ECS AND WAS ASSOCIATED WITH ABERRANT HYPERMETHYLATION AT THE ENOS PROXIMAL PROMOTER (N=7). REPORTER GENE SILENCING WAS INDEPENDENT OF TRANSGENE COPY NUMBER AND INTEGRATION POSITION, INDICATING THAT THE SSRE IS A CRITICAL CIS-ELEMENT NECESSARY FOR ENOS TRANSCRIPTION IN VIVO. THE KLF MUTATION DEMONSTRATED AN INTEGRATION SITE-SPECIFIC DECREASE IN ENOS TRANSCRIPTION, AGAIN WITH MARKED PROMOTER METHYLATION (N=8), SUGGESTING THAT THE SSRE ALONE IS NOT SUFFICIENT FOR ENOS TRANSCRIPTION IN VIVO. IN WILD TYPE MICE, THE NATIVE ENOS PROMOTER WAS SIGNIFICANTLY HYPERMETHYLATED IN ECS FROM THE ATHEROPRONE REGIONS WHERE ENOS EXPRESSION WAS MARKEDLY REPRESSED BY CHRONIC DISTURBED FLOW, DEMONSTRATING THAT ENOS EXPRESSION IS REGULATED BY FLOW-DEPENDENT DNA METHYLATION THAT IS REGION-SPECIFIC IN THE ARTERIAL ENDOTHELIUM IN VIVO. CONCLUSIONS: WE REPORT, FOR THE FIRST TIME, THAT THE SSRE AND KLF ELEMENTS ARE CRITICAL FLOW SENSORS NECESSARY FOR A TRANSCRIPTIONALLY PERMISSIVE, HYPOMETHYLATED ENOS PROMOTER IN ECS UNDER CHRONIC SHEAR STRESS IN VIVO. MOREOVER, ENOS EXPRESSION IS REGULATED BY FLOW-DEPENDENT EPIGENETIC MECHANISMS, WHICH OFFERS NOVEL MECHANISTIC INSIGHT ON ENOS GENE REGULATION IN ATHEROGENESIS. 2021 17 6516 35 TRANSCRIPTIONAL AND EPIGENETIC FACTORS ASSOCIATED WITH EARLY THROMBOSIS OF FEMORAL ARTERY INVOLVED IN ARTERIOVENOUS FISTULA. ARTERIOVENOUS FISTULAS (AVFS), CREATED FOR HEMODIALYSIS IN END-STAGE RENAL DISEASE PATIENTS, MATURE THROUGH THE OUTWARD REMODELING OF THE OUTFLOW VEIN. HOWEVER, EARLY THROMBOSIS AND CHRONIC INFLAMMATION ARE DETRIMENTAL TO THE PROCESS OF AVF MATURATION AND PRECIPITATE AVF MATURATION FAILURE. FOR THE SUCCESSFUL REMODELING OF THE OUTFLOW VEIN, BLOOD FLOW THROUGH THE FISTULA IS ESSENTIAL, BUT EARLY ARTERIAL THROMBOSIS ATTENUATES THIS BLOOD FLOW, AND THE VESSELS BECOME THROMBOSED AND STENOSED, LEADING TO AVF FAILURE. THE ALTERED EXPRESSION OF VARIOUS PROTEINS INVOLVED IN MAINTAINING VESSEL PATENCY OR THROMBOSIS IS REGULATED BY GENES OF WHICH THE EXPRESSION IS REGULATED BY TRANSCRIPTION FACTORS AND MICRORNAS. IN THIS STUDY, USING THROMBOSED AND STENOSED ARTERIES FOLLOWING AVF CREATION, WE DELINEATED TRANSCRIPTION FACTORS AND MICRORNAS ASSOCIATED WITH DIFFERENTIALLY EXPRESSED GENES IN BULK RNA SEQUENCING DATA USING UPSTREAM AND CAUSAL NETWORK ANALYSIS. WE OBSERVED CHANGES IN MANY TRANSCRIPTION FACTORS AND MICRORNAS THAT ARE INVOLVED IN ANGIOGENESIS; VASCULAR SMOOTH MUSCLE CELL PROLIFERATION, MIGRATION, AND PHENOTYPIC CHANGES; ENDOTHELIAL CELL FUNCTION; HYPOXIA; OXIDATIVE STRESS; VESSEL REMODELING; IMMUNE RESPONSES; AND INFLAMMATION. THESE FACTORS AND MICRORNAS PLAY A CRITICAL ROLE IN THE UNDERLYING MOLECULAR MECHANISMS IN AVF MATURATION. WE ALSO OBSERVED EPIGENETIC FACTORS INVOLVED IN GENE REGULATION ASSOCIATED WITH THESE MOLECULAR MECHANISMS. THE RESULTS OF THIS STUDY INDICATE THE IMPORTANCE OF INVESTIGATING THE TRANSCRIPTIONAL AND EPIGENETIC REGULATION OF AVF MATURATION AND MATURATION FAILURE AND TARGETING FACTORS PRECIPITATING EARLY THROMBOSIS AND STENOSIS. 2022 18 4303 32 MICRORNA-223 INHIBITS TISSUE FACTOR EXPRESSION IN VASCULAR ENDOTHELIAL CELLS. OBJECTIVE: ATHEROSCLEROSIS IS A CHRONIC INFLAMMATORY PROCESS, IN WHICH VASCULAR ENDOTHELIAL CELLS (ECS) BECOME DYSFUNCTIONAL OWING TO THE EFFECTS OF CHEMICAL SUBSTANCES, SUCH AS INFLAMMATORY FACTOR AND GROWTH FACTORS. TISSUE FACTOR (TF) EXPRESSION IS INDUCED BY THE ABOVE CHEMICAL SUBSTANCES IN ACTIVATED ECS. TF INITIATES THROMBOSIS ON DISRUPTED ATHEROSCLEROTIC PLAQUES WHICH PLAYS AN ESSENTIAL ROLE DURING THE ONSET OF ACUTE CORONARY SYNDROMES (ACS). INCREASING EVIDENCES SUGGEST THE IMPORTANT ROLE OF MICRORNAS AS EPIGENETIC REGULATORS OF ATHEROSCLEROTIC DISEASE. THE AIM OF OUR STUDY IS TO IDENTIFY IF MICRORNA-223 (MIR-223) TARGETS TF IN ECS. METHODS AND RESULTS: BIOINFORMATIC ANALYSIS SHOWED THAT TF IS A TARGET CANDIDATE OF MIR-223. WESTERN BLOTTING ANALYSIS REVEALED THAT TUMOR NECROSIS FACTOR ALPHA (TNF-ALPHA) INCREASED TF EXPRESSION IN AORTA OF C57BL/6J MICE AND CULTURED ECS (EA.HY926 CELLS AND HUVEC) AFTER 4 H TREATMENT. IN TNF-ALPHA TREATED ECS, TF MRNA WAS ALSO INCREASED MEASURED BY REAL-TIME PCR. REAL-TIME PCR RESULTS SHOWED THAT MIR-223 LEVELS WERE DOWNREGULATED IN TNF-ALPHA-TREATED AORTA OF C57BL/6J MICE AND CULTURED ECS. TRANSFECTION OF ECS WITH MIR-223 MIMIC OR MIR-223 INHIBITOR MODIFIED TF EXPRESSION BOTH IN MRNA AND PROTEIN LEVELS. LUCIFERASE ASSAYS CONFIRMED THAT MIR-223 SUPPRESSED TF EXPRESSION BY BINDING TO THE SEQUENCE OF TF 3'-UNTRANSLATED REGIONS (3'UTR). TF PROCOAGULANT ACTIVITY WAS INHIBITED BY OVEREXPRESSING MIR-223 WITH OR WITHOUT TNF-ALPHA STIMULATION. CONCLUSIONS: MIR-223-MEDIATED SUPPRESSION OF TF EXPRESSION PROVIDES A NOVEL MOLECULAR MECHANISM FOR THE REGULATION OF COAGULATION CASCADE, AND SUGGESTS A CLUE AGAINST THROMBOGENESIS DURING THE PROCESS OF ATHEROSCLEROTIC PLAQUE RUPTURE. 2014 19 1092 29 COHESIN MUTATIONS IN MYELOID MALIGNANCIES. COHESIN IS A MULTISUBUNIT PROTEIN COMPLEX THAT FORMS A RING-LIKE STRUCTURE AROUND DNA. IT IS ESSENTIAL FOR SISTER CHROMATID COHESION, CHROMATIN ORGANIZATION, TRANSCRIPTIONAL REGULATION, AND DNA DAMAGE REPAIR AND PLAYS A MAJOR ROLE IN DYNAMICALLY SHAPING THE GENOME ARCHITECTURE AND MAINTAINING DNA INTEGRITY. THE CORE COMPLEX SUBUNITS STAG2, RAD21, SMC1, AND SMC3, AS WELL AS ITS MODULATORS PDS5A/B, WAPL, AND NIPBL, HAVE BEEN FOUND TO BE RECURRENTLY MUTATED IN HEMATOLOGIC AND SOLID MALIGNANCIES. THESE MUTATIONS ARE FOUND ACROSS THE FULL SPECTRUM OF MYELOID NEOPLASIA, INCLUDING PEDIATRIC DOWN SYNDROME-ASSOCIATED ACUTE MEGAKARYOBLASTIC LEUKEMIA, MYELODYSPLASTIC SYNDROMES, CHRONIC MYELOMONOCYTIC LEUKEMIA, AND DE NOVO AND SECONDARY ACUTE MYELOID LEUKEMIAS. THE MECHANISMS BY WHICH COHESIN MUTATIONS ACT AS DRIVERS OF CLONAL EXPANSION AND DISEASE PROGRESSION ARE STILL POORLY UNDERSTOOD. RECENT STUDIES HAVE DESCRIBED THE IMPACT OF COHESIN ALTERATIONS ON SELF-RENEWAL AND DIFFERENTIATION OF HEMATOPOIETIC STEM AND PROGENITOR CELLS, WHICH ARE ASSOCIATED WITH CHANGES IN CHROMATIN AND EPIGENETIC STATE DIRECTING LINEAGE COMMITMENT, AS WELL AS GENOMIC INTEGRITY. HEREIN, WE REVIEW THE ROLE OF THE COHESIN COMPLEX IN HEALTHY AND MALIGNANT HEMATOPOIESIS. WE DISCUSS CLINICAL IMPLICATIONS OF COHESIN MUTATIONS IN MYELOID MALIGNANCIES AND DISCUSS OPPORTUNITIES FOR THERAPEUTIC TARGETING. 2021 20 3719 26 INHIBITION OF BET PROTEINS REDUCES RIGHT VENTRICLE HYPERTROPHY AND PULMONARY HYPERTENSION RESULTING FROM COMBINED HYPOXIA AND PULMONARY INFLAMMATION. PULMONARY HYPERTENSION IS A CO-MORBIDITY, WHICH STRONGLY PARTICIPATES IN MORBI-MORTALITY IN PATIENTS WITH CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD). RECENT FINDINGS SHOWED THAT BROMODOMAIN-CONTAINING PROTEINS, IN CHARGE OF READING HISTONE ACETYLATION, COULD BE INVOLVED IN PULMONARY ARTERIAL HYPERTENSION. OUR AIM WAS TO STUDY THE EFFECT OF I-BET151, AN INHIBITOR OF BROMODOMAIN AND EXTRA-TERMINAL DOMAIN (BET), ON THE RIGHT VENTRICLE HYPERTROPHY AND PULMONARY HYPERTENSION, INDUCED BY A COMBINATION OF CHRONIC HYPOXIA AND PULMONARY INFLAMMATION, AS THE TWO MAIN STIMULI ENCOUNTERED IN COPD. ADULT WISTAR MALE RATS, EXPOSED TO CHRONIC HYPOXIA PLUS PULMONARY INFLAMMATION (CHPI), SHOWED A SIGNIFICANT RIGHT VENTRICLE HYPERTROPHY (+57%, P < 0.001), AN INCREASE IN SYSTOLIC PRESSURE (+46%, P < 0.001) AND IN CONTRACTION SPEED (+36%, P < 0.001), WHEN COMPARED TO CONTROL ANIMALS. I-BET151 TREATED ANIMALS (CHPI-IB) SHOWED RESTORED HEMODYNAMIC PARAMETERS TO LEVELS SIMILAR TO CONTROL ANIMALS, DESPITE CHRONIC HYPOXIA PLUS EXPOSURE TO PULMONARY INFLAMMATION. THEY DISPLAYED LOWER RIGHT VENTRICLE HYPERTROPHY AND HEMATOCRIT COMPARED TO THE CHPI GROUP (RESPECTIVELY -16%, P < 0.001; AND -9%, P < 0.05). OUR DESCRIPTIVE STUDY SHOWS A VALUABLE EFFECT OF THE INHIBITION OF BROMODOMAIN AND EXTRA-TERMINAL DOMAIN PROTEINS ON HEMODYNAMIC PARAMETERS, DESPITE THE PRESENCE OF CHRONIC HYPOXIA AND PULMONARY INFLAMMATION. THIS SUGGESTS THAT SUCH INHIBITION COULD BE OF POTENTIAL INTEREST FOR COPD PATIENTS WITH PULMONARY HYPERTENSION. FURTHER STUDIES ARE NEEDED TO UNRAVEL THE UNDERLYING MECHANISMS INVOLVED AND THE NET BENEFITS OF INHIBITING ADAPTATIONS TO CHRONIC HYPOXIA. 2018