1 3724 144 INHIBITION OF EZH2 AMELIORATES CARTILAGE ENDPLATE DEGENERATION AND ATTENUATES THE PROGRESSION OF INTERVERTEBRAL DISC DEGENERATION VIA DEMETHYLATION OF SOX-9. BACKGROUND: CARTILAGINOUS ENDPLATE (CEP) DEGENERATION IS CONSIDERED AS ONE OF THE MAJOR CAUSES OF INTERVERTEBRAL DISC DEGENERATION (IVDD) WHICH CAUSES LOW BACK PAIN. RECENT STUDIES HAVE PROVED THAT EPIGENETIC ALTERATION IS INVOLVED IN A VARIETY OF DISEASES. THIS WORK EXPLORED THE ROLE OF HISTONE METHYLTRANSFERASE ENHANCER OF ZESTE HOMOLOGUE 2 (EZH2) IN CEP DEGENERATION, AS WELL AS ITS UNDERLYING EPIGENETIC MECHANISMS, AND CONFIRMED THE EFFECT OF EZH2 KNOCKDOWN ON DELAYING IVDD DEVELOPMENT. METHODS: WESTERN BLOTTING, IMMUNOFLUORESCENCE STAINING, AND CHIP ASSAY WERE APPLIED TO DEMONSTRATE THE MOLECULAR MECHANISM OF EZH2 IN CEP TISSUE. THE THERAPEUTIC POTENTIAL OF EZH2 WAS INVESTIGATED USING PUNCTURE-INDUCED RAT MODELS. FINDINGS: THE EZH2 EXPRESSION WAS UPREGULATED IN HUMAN AND RAT CEP TISSUE. IT WAS ALSO FOUND THAT THE OVEREXPRESSION OF EZH2 SUPPRESSED THE EXPRESSION OF COLLAGEN II, AGGRECAN AND SOX-9, AND PROMOTED THE EXPRESSION OF ADTAMTS5 AND MMP13 IN RAT ENDPLATE CHONDROCYTES (EPCS), WHICH COULD BE REVERSED BY EZH2 SILENCING. THE CORRELATION BETWEEN EZH2 AND SOX-9 WAS FURTHER EXPLORED, WHILE OVEREXPRESSION OF SOX-9 COULD REVERSE THE EFFECT OF EZH2 IN RAT EPCS. MOREOVER, INHIBITION OF EZH2 UPREGULATED THE LEVEL OF SOX-9 BY DEMETHYLATING H3K27ME3 AT SOX-9 PROMOTER SITES, REVEALING THE REGULATORY MECHANISM OF EZH2 ON SOX-9. MEANWHILE, PUNCTURE-INDUCED RAT MODELS SHOWED THAT EZH2 KNOCKDOWN EXERTED A PROTECTIVE EFFECT ON CEP AND DISC DEGENERATION. INTERPRETATION: THIS STUDY REVEALS THAT EZH2 INHIBITION IS A PROMISING STRATEGY FOR MITIGATING THE SYMPTOMS AND PROGRESSION OF IVDD. FUNDING: THIS STUDY WAS FUNDED BY THE NATURAL SCIENCE FOUNDATION OF ZHEJIANG PROVINCE (Y16H060034). AUTHORS DECLARE THAT THE FUNDERS HAD NO INVOLVEMENT IN THE STUDY DESIGN, DATA ANALYSIS AND INTERPRETATION OF THE RESULTS. 2019 2 5068 31 PHYSICAL ACTIVITY AND PROGENITOR CELL-MEDIATED ENDOTHELIAL REPAIR IN CHRONIC HEART FAILURE: IS THERE A ROLE FOR EPIGENETICS? CHRONIC HEART FAILURE (CHF) IS THE MOST COMMON CARDIAC DISEASE AMONG THE ELDERLY AND A LEADING CAUSE OF MORTALITY IN ELDERLY PATIENTS. ENDOTHELIAL DYSFUNCTION IS HELD TO HAVE A MAJOR ROLE IN THE DEVELOPMENT AND PROGRESSION OF CHF, WHICH RESULTS IN PROGRESSIVELY IMPAIRED FUNCTIONAL CAPACITY. ENDOTHELIAL PROGENITOR CELLS (EPCS) AND CIRCULATING ANGIOGENIC CELLS (CACS) ARE THE MAIN PLAYERS INVOLVED IN THE ENDOGENOUS REPAIR MECHANISMS THAT CAN COUNTERACT ENDOTHELIAL DYSFUNCTION. A MOUNTING BODY OF DATA INDICATES THAT EXERCISE ENHANCES ENDOTHELIAL RENEWAL THROUGH MOBILIZATION OF BONE MARROW-DERIVED EPCS AND CACS, MAKING IT AN EFFECTIVE THERAPEUTIC TOOL FOR CHF. INTERESTINGLY, EMERGING EVIDENCE HAS BEEN SHOWING THAT EXERCISE TRAINING CAN ALSO PROMOTE EPIGENETIC MODIFICATIONS, E.G. DNA METHYLATION, HISTONE MODIFICATIONS, AND DIFFERENTIAL EXPRESSION OF SPECIFIC NON-CODING RNAS LIKE MICRORNA (MIRNAS). SINCE DEREGULATION OF THE MIRNAS INVOLVED IN ENDOTHELIAL FUNCTION MODULATION HAS WIDELY BEEN DOCUMENTED IN CIRCULATING CELLS AND PLASMA OF CHF PATIENTS, DEREGULATION OF EPIGENETIC FEATURES COULD PLAY A KEY ROLE IN DISEASE PROGRESSION. HERE, WE REVIEW CURRENT KNOWLEDGE OF THE CONTRIBUTION OF EPCS AND CACS TO ENDOTHELIAL REPAIR MECHANISMS IN CHF PATIENTS, FOCUSING ON THE EFFECTS INDUCED BY EXERCISE TRAINING AND HYPOTHESIZING THAT SOME OF THESE EFFECTS CAN BE MEDIATED BY EPIGENETIC MECHANISMS. 2016 3 6413 18 THE STATE OF ART OF REGENERATIVE THERAPY IN CARDIOVASCULAR ISCHEMIC DISEASE: BIOLOGY, SIGNALING PATHWAYS, AND EPIGENETICS OF ENDOTHELIAL PROGENITOR CELLS. ISCHEMIC HEART DISEASE IS CURRENTLY A MAJOR CAUSE OF MORTALITY AND MORBIDITY WORLDWIDE. NEVERTHELESS, THE ACTUAL THERAPEUTIC SCENARIO DOES NOT TARGET MYOCARDIAL CELL REGENERATION AND CONSEQUENTLY, THE PROGRESSION TOWARD THE LATE STAGE OF CHRONIC HEART FAILURE IS COMMON. ENDOTHELIAL PROGENITOR CELLS (EPCS) ARE BONE MARROW-DERIVED STEM CELLS THAT CONTRIBUTE TO THE HOMEOSTASIS OF THE ENDOTHELIAL WALL IN ACUTE AND CHRONIC ISCHEMIC DISEASE. CALCIUM MODULATION AND OTHER MOLECULAR PATHWAYS (NOTCH, VEGFR, AND CXCR4) CONTRIBUTE TO EPC PROLIFERATION AND DIFFERENTIATION. THE PRESENT REVIEW PROVIDES A SUMMARY OF EPC BIOLOGY WITH A PARTICULAR FOCUS ON THE REGULATORY PATHWAYS OF EPCS AND DESCRIBES PROMISING APPLICATIONS FOR CARDIOVASCULAR CELL THERAPY. 2020 4 328 35 ALPHA-KETOGLUTARIC ACID AMELIORATES INTERVERTEBRAL DISC DEGENERATION BY BLOCKING THE IL-6/JAK2/STAT3 PATHWAY. INTERVERTEBRAL DISC DEGENERATION (IVDD) IS THE MAJOR CAUSE OF LOW BACK PAIN. ALPHA-KETOGLUTARIC ACID (ALPHA-KG), AN IMPORTANT INTERMEDIATE IN ENERGY METABOLISM, HAS VARIOUS FUNCTIONS, INCLUDING EPIGENETIC REGULATION, MAINTENANCE OF REDOX HOMEOSTASIS, AND ANTI-AGING, BUT WHETHER IT CAN AMELIORATE IVDD HAS NOT BEEN REPORTED. HERE, WE EXAMINED THE IMPACTS OF LONG-TERM ADMINISTRATION OF A-KG ON AGING-ASSOCIATED IVDD IN ADULT RATS. IN VIVO AND IN VITRO EXPERIMENTS SHOWED THAT ALPHA-KG SUPPLEMENTATION EFFECTIVELY AMELIORATED IVDD IN RATS AND THE SENESCENCE OF NUCLEUS PULPOSUS CELLS (NPCS). ALPHA-KG SUPPLEMENTATION SIGNIFICANTLY ATTENUATED SENESCENCE, APOPTOSIS AND MMP-13 PROTEIN EXPRESSION, AND IT INCREASED THE SYNTHESIS OF AGGRECAN AND COLLAGEN II IN IL-1BETA-TREATED NPCS. IN ADDITION, ALPHA-KG SUPPLEMENTATION REDUCED THE LEVELS OF IL-6, PHOSPHORYLATED JAK2 AND STAT3, AND THE NUCLEAR TRANSLOCATION OF P-STAT3 IN IL-1BETA-INDUCED DEGENERATING NPCS. THE EFFECTS OF ALPHA-KG WERE ENHANCED BY AG490 IN NPCS. THE UNDERLYING MECHANISM MAY INVOLVE THE INHIBITION OF JAK2/STAT3 PHOSPHORYLATION AND THE REDUCTION OF IL-6 EXPRESSION. OUR FINDINGS MAY HELP IN THE DEVELOPMENT OF NEW THERAPEUTIC STRATEGIES FOR IVDD. 2023 5 2284 16 EPIGENETIC REGULATION IN INTERVERTEBRAL DISC DEGENERATION. INTERVERTEBRAL DISC (IVD) DEGENERATION IS THE LEADING CAUSE OF LOW BACK PAIN, WHICH HAS A STRIKING IMPACT ON NUMEROUS PATIENTS. THEREFORE, COMPREHENSIVELY ILLUMINATING THE REGULATORY MECHANISMS OF IVD DEGENERATION IS OF GREAT SIGNIFICANCE. HERE, WE DISCUSS THE LATEST ADVANCES IN UNDERSTANDING THE MAIN EPIGENETIC MECHANISMS REGULATING IVD DEGENERATION. 2022 6 5740 39 SMOKING AND TETRAMER TRYPTASE ACCELERATE INTERVERTEBRAL DISC DEGENERATION BY INDUCING METTL14-MEDIATED DIXDC1 M(6) MODIFICATION. ALTHOUGH CIGARETTE SMOKING (CS) AND LOW BACK PAIN (LBP) ARE COMMON WORLDWIDE, THEIR CORRELATIONS AND THE MECHANISMS OF ACTION REMAIN UNCLEAR. WE HAVE SHOWN THAT EXCESSIVE ACTIVATION OF MAST CELLS (MCS) AND THEIR PROTEASES PLAY KEY ROLES IN CS-ASSOCIATED DISEASES, LIKE ASTHMA, CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD), BLOOD COAGULATION, AND LUNG CANCER. PREVIOUS STUDIES HAVE ALSO SHOWN THAT MCS AND THEIR PROTEASES INDUCE DEGENERATIVE MUSCULOSKELETAL DISEASE. BY USING A CUSTOM-DESIGNED SMOKE-EXPOSURE MOUSE SYSTEM, WE DEMONSTRATED THAT CS RESULTS IN INTERVERTEBRAL DISC (IVD) DEGENERATION AND RELEASE OF MC-RESTRICTED TETRAMER TRYPTASES (TTS) IN THE IVDS. TTS WERE FOUND TO REGULATE THE EXPRESSION OF METHYLTRANSFERASE 14 (METTL14) AT THE EPIGENETIC LEVEL BY INDUCING N6-METHYLADENOSINE (M(6)A) DEPOSITION IN THE 3' UNTRANSLATED REGION (UTR) OF THE TRANSCRIPT THAT ENCODES DISHEVELLED-AXIN (DIX) DOMAIN-CONTAINING 1 (DIXDC1). THAT REACTION INCREASES THE MRNA STABILITY AND EXPRESSION OF DIXDC1. DIXDC1 FUNCTIONALLY INTERACTS WITH DISRUPTED IN SCHIZOPHRENIA 1 (DISC1) TO ACCELERATE THE DEGENERATION AND SENESCENCE OF NUCLEUS PULPOSUS (NP) CELLS BY ACTIVATING A CANONICAL WNT PATHWAY. OUR STUDY DEMONSTRATES THE ASSOCIATION BETWEEN CS, MC-DERIVED TTS, AND LBP. THESE FINDINGS RAISE THE POSSIBILITY THAT METTL14-MEDICATED DIXDC1 M(6)A MODIFICATION COULD SERVE AS A POTENTIAL THERAPEUTIC TARGET TO BLOCK THE DEVELOPMENT OF DEGENERATION OF THE NP IN LBP PATIENTS. 2023 7 4033 48 M6A HYPOMETHYLATION OF DNMT3B REGULATED BY ALKBH5 PROMOTES INTERVERTEBRAL DISC DEGENERATION VIA E4F1 DEFICIENCY. BACKGROUND: THE INTERVERTEBRAL DISC (IVD) DEGENERATION IS THE LEADING CAUSE OF LOW BACK PAIN, WHICH ACCOUNTS FOR A MAIN CAUSE OF DISABILITY. N6-METHYLADENOSINE (M6A) IS THE MOST ABUNDANT INTERNAL MODIFICATION IN EUKARYOTIC MESSENGER RNAS AND IS INVOLVED IN VARIOUS DISEASES AND CELLULAR PROCESSES BY MODULATING MRNA FATE. HOWEVER, THE CRITICAL ROLE OF M6A REGULATION IN IVD DEGENERATION REMAINS UNCLEAR. NUCLEUS PULPOSUS CELL (NPC) SENESCENCE IS CRITICAL FOR THE PROGRESSION OF IVD DEGENERATION. HERE, WE UNCOVERED THE ROLE AND EXPLORED THE REGULATORY MECHANISM OF M6A IN NPC SENESCENCE DURING IVD DEGENERATION. METHODS: IDENTIFICATION OF NPC SENESCENCE DURING IVD DEGENERATION WAS BASED ON THE ANALYSIS OF TISSUE SAMPLES AND THE CELLULAR MODEL. ALKBH5 UPREGULATION INDUCING CELLULAR SENESCENCE WAS CONFIRMED BY FUNCTIONAL EXPERIMENTS IN VIVO AND IN VITRO. CHIP-QPCR AND DNA-PULLDOWN WERE USED TO REVEAL INCREASED ALKBH5 WAS REGULATED BY KDM4A-MEDIATED H3K9ME3. FURTHERMORE, ME-RIP-SEQ WAS PERFORMED TO IDENTIFY M6A HYPOMETHYLATION OF DNMT3B TRANSCRIPTS IN SENESCENT NPCS. STABILITY ANALYSIS SHOWED THAT DNMT3B EXPRESSION WAS ENHANCED FOR LESS YTHDF2 RECOGNITION AND INCREASED DNMT3B PROMOTED NPC SENESCENCE AND IVD DEGENERATION VIA E4F1 METHYLATION BY IN VIVO AND IN VITRO ANALYSES. RESULTS: EXPRESSION OF ALKBH5 IS ENHANCED DURING IVD DEGENERATION AND NPC SENESCENCE, DUE TO DECREASED KDM4A-MEDIATED H3K9ME3 MODIFICATION. FUNCTIONALLY, ALKBH5 CAUSES NPC SENESCENCE BY DEMETHYLATING DNMT3B TRANSCRIPTS AND IN TURN PROMOTING ITS EXPRESSION VIA LESS YTHDF2 RECOGNITION AND FOLLOWING DEGRADATION DUE TO TRANSCRIPT HYPOMETHYLATION IN VITRO AND IN VIVO. INCREASED DNMT3B PROMOTES THE DEVELOPMENT OF IVD DEGENERATION AND NPC SENESCENCE, MECHANISTICALLY BY METHYLATING CPG ISLANDS OF E4F1 AT THE PROMOTER REGION AND THUS RESTRAINING ITS TRANSCRIPTION AND EXPRESSION. CONCLUSIONS: COLLECTIVELY, OUR FINDINGS REVEAL AN EPIGENETIC INTERPLAY MECHANISM IN NPC SENESCENCE AND IVD DEGENERATION, PRESENTING A CRITICAL PRO-SENESCENCE ROLE OF ALKBH5 AND M6A HYPOMETHYLATION, HIGHLIGHTING THE THERAPEUTIC POTENTIAL OF TARGETING THE M6A/DNMT3B/E4F1 AXIS FOR TREATING IVD DEGENERATION. 2022 8 4356 42 MIR-30A-5P PROMOTES GLOMERULAR PODOCYTE APOPTOSIS VIA DNMT1-MEDIATED HYPERMETHYLATION UNDER HYPERHOMOCYSTEINEMIA. ABNORMAL ELEVATION OF HOMOCYSTEINE (HCY) LEVEL IS CLOSELY RELATED TO THE DEVELOPMENT AND PROGRESSION OF CHRONIC KIDNEY DISEASE (CKD), WITH THE MOLECULAR MECHANISMS THAT ARE NOT FULLY ELUCIDATED. GIVEN THE DEMONSTRATION THAT MIR-30A-5P IS SPECIFICALLY EXPRESSED IN GLOMERULAR PODOCYTES, IN THE PRESENT STUDY WE AIMED TO INVESTIGATE THE ROLE AND POTENTIAL UNDERLYING MECHANISM OF MIR-30A-5P IN GLOMERULAR PODOCYTE APOPTOSIS INDUCED BY HCY. WE FOUND THAT ELEVATED HCY DOWNREGULATES MIR-30A-5P EXPRESSION IN THE MICE AND HCY-TREATED PODOCYTES, AND MIR-30A-5P DIRECTLY TARGETS THE 3'-UNTRANSLATED REGION (3'-UTR) OF THE FORKHEAD BOX A1 (FOXA1) AND OVEREXPRESSION OF MIR-30A-5P INHIBITS FOXA1 EXPRESSION. BY NMS-PCR AND MASSARRAY QUANTITATIVE METHYLATION ANALYSIS, WE SHOWED THE INCREASED DNA METHYLATION LEVEL OF MIR-30A-5P PROMOTER BOTH AND . MEANWHILE, DUAL-LUCIFERASE REPORTER ASSAY SHOWED THAT THE REGION BETWEEN --1400 AND --921 BP OF MIR-30A-5P PROMOTER IS A POSSIBLE REGULATORY ELEMENT FOR ITS TRANSCRIPTION. MECHANISTIC STUDIES INDICATED THAT DNA METHYLTRANSFERASE ENZYME 1 (DNMT1) IS THE KEY REGULATOR OF MIR-30A-5P, WHICH IN TURN ENHANCES MIR-30A-5P PROMOTER METHYLATION LEVEL AND THEREBY INHIBITS ITS EXPRESSION. TAKEN TOGETHER, OUR RESULTS REVEALED THAT EPIGENETIC MODIFICATION OF MIR-30A-5P IS INVOLVED IN GLOMERULAR PODOCYTE INJURY INDUCED BY HCY, PROVIDING A DIAGNOSTIC MARKER CANDIDATE AND NOVEL THERAPEUTIC TARGET IN CKD INDUCED BY HCY. 2022 9 5636 47 SERELAXIN ALLEVIATES CARDIAC FIBROSIS THROUGH INHIBITING ENDOTHELIAL-TO-MESENCHYMAL TRANSITION VIA RXFP1. RATIONALE: CARDIAC FIBROSIS IS AN INTEGRAL CONSTITUENT OF EVERY FORM OF CHRONIC HEART DISEASE, AND PERSISTENCE OF FIBROSIS REDUCES TISSUE COMPLIANCE AND ACCELERATES THE PROGRESSION TO HEART FAILURE. RELAXIN-2 IS A HUMAN HORMONE, WHICH HAS VARIOUS PHYSIOLOGICAL FUNCTIONS SUCH AS MEDIATING RENAL VASODILATION IN PREGNANCY. ITS RECOMBINANT FORM SERELAXIN HAS RECENTLY BEEN TESTED IN CLINICAL TRIALS AS A THERAPY FOR ACUTE HEART FAILURE BUT DID NOT MEET ITS PRIMARY ENDPOINTS. THE AIM OF THIS STUDY IS TO EXAMINE WHETHER SERELAXIN HAS AN ANTI-FIBROTIC EFFECT IN THE HEART AND THEREFORE COULD BE BENEFICIAL IN CHRONIC HEART FAILURE. METHODS: WE UTILIZED TWO DIFFERENT CARDIAC FIBROSIS MOUSE MODELS (ASCENDING AORTIC CONSTRICTION (AAC) AND ANGIOTENSIN II (ATII) ADMINISTRATION VIA OSMOTIC MINIPUMPS) TO ASSESS THE ANTI-FIBROTIC POTENTIAL OF SERELAXIN. HISTOLOGICAL ANALYSIS, IMMUNOFLUORESCENCE STAINING AND MOLECULAR ANALYSIS WERE PERFORMED TO ASSESS THE FIBROSIS LEVEL AND INDICATE ENDOTHELIAL CELLS WHICH ARE UNDERGOING ENDMT. IN VITRO TGFBETA1-INDUCED ENDOTHELIAL-TO-MESENCHYMAL TRANSITION (ENDMT) ASSAYS WERE PERFORMED IN HUMAN CORONARY ARTERY ENDOTHELIAL CELLS AND MOUSE CARDIAC ENDOTHELIAL CELLS (MCECS) AND WERE EXAMINED USING MOLECULAR METHODS. CHROMATIN IMMUNOPRECIPITATION-QPCR ASSAY WAS UTILIZED TO IDENTIFY THE SERELAXIN EFFECT ON CHROMATIN REMODELING IN THE RXFP1 PROMOTER REGION IN MCECS. RESULTS: OUR RESULTS DEMONSTRATE A SIGNIFICANT AND DOSE-DEPENDENT ANTI-FIBROTIC EFFECT OF SERELAXIN IN THE HEART IN BOTH MODELS. WE FURTHER SHOW THAT SERELAXIN MEDIATES THIS EFFECT, AT LEAST IN PART, THROUGH INHIBITION OF ENDMT THROUGH THE ENDOTHELIAL RELAXIN FAMILY PEPTIDE RECEPTOR 1 (RXFP1). WE FURTHER DEMONSTRATE THAT SERELAXIN ADMINISTRATION IS ABLE TO INCREASE ITS OWN RECEPTOR EXPRESSION (RXFP1) THROUGH EPIGENETIC REGULATION IN FORM OF HISTONE MODIFICATIONS BY ATTENUATING TGFBETA-PSMAD2/3 SIGNALING IN ENDOTHELIAL CELLS. CONCLUSIONS: THIS STUDY IS THE FIRST TO IDENTIFY THAT SERELAXIN INCREASES THE EXPRESSION OF ITS OWN RECEPTOR RXFP1 AND THAT THIS MEDIATES THE INHIBITION OF ENDMT AND CARDIAC FIBROSIS, SUGGESTING THAT SERELAXIN MAY HAVE A BENEFICIAL EFFECT AS ANTI-FIBROTIC THERAPY IN CHRONIC HEART FAILURE. 2020 10 3388 33 HOMOCYSTEINE INDUCES PODOCYTE APOPTOSIS BY REGULATING MIR-1929-5P EXPRESSION THROUGH C-MYC, DNMT1 AND EZH2. CHRONIC KIDNEY DISEASE (CKD) IS A COMMON AND COMPLEX DISEASE IN KIDNEYS WHICH HAS BEEN ASSOCIATED WITH AN INCREASED RISK OF RENAL CELL CARCINOMA. ELEVATED HOMOCYSTEINE (HCY) LEVELS ARE KNOWN TO INFLUENCE THE DEVELOPMENT AND PROGRESSION OF CKD BY REGULATING PODOCYTE INJURY AND APOPTOSIS. TO INVESTIGATE THE MOLECULAR MECHANISMS TRIGGERED IN PODOCYTES BY HCY, WE USED CBS(+/-) MICE AND OBSERVED THAT HIGHER HCY LEVELS INCREASED THE APOPTOSIS RATE OF PODOCYTES WITH ACCOMPANYING GLOMERULAR DAMAGE. HCY-INDUCED PODOCYTE INJURY AND APOPTOSIS IN CBS(+/-) MICE WAS REGULATED BY INHIBITION OF MICRORNA (MIR)-1929-5P EXPRESSION. OVEREXPRESSION OF MIR-1929-5P IN PODOCYTES INHIBITED APOPTOSIS BY UPREGULATING BCL-2. FURTHERMORE, THE EXPRESSION OF MIR-1929-5P WAS REGULATED BY EPIGENETIC MODIFICATIONS OF ITS PROMOTER. HCY UPREGULATED DNA METHYLTRANSFERASE 1 (DNMT1) AND ENHANCER OF ZESTE HOMOLOG 2 (EZH2) LEVELS, RESULTING IN INCREASED DNA METHYLATION AND H3K27ME3 LEVELS ON THE MIR-1929-5P PROMOTER. ADDITIONALLY, WE OBSERVED THAT C-MYC RECRUITED DNMT1 AND EZH2 TO THE MIR-1929-5P PROMOTER AND SUPPRESSED THE EXPRESSION OF MIR-1929-5P. IN SUMMARY, WE DEMONSTRATED THAT HCY PROMOTES PODOCYTE APOPTOSIS THROUGH THE REGULATION OF THE EPIGENETIC MODIFIERS DNMT1 AND EZH2, WHICH ARE RECRUITED BY C-MYC TO THE PROMOTER OF MIR-1929-5P TO SILENCE MIR-1929-5P EXPRESSION. 2021 11 273 43 AGE-INDUCED SUPPRESSION OF EZH2 MEDIATES INJURY OF PODOCYTES BY REDUCING H3K27ME3. BACKGROUND: CHRONIC HYPERGLYCEMIA, A PIVOTAL FEATURE OF DIABETES MELLITUS (DM), INITIATES THE FORMATION OF ADVANCED GLYCATION END PRODUCTS (AGES) AND THE DYSREGULATION OF EPIGENETIC MECHANISMS, WHICH MAY CAUSE INJURY TO RENAL PODOCYTES, A CENTRAL FEATURE OF DIABETIC KIDNEY DISEASE (DKD). PREVIOUS DATA OF OUR GROUP SHOWED THAT AGES SIGNIFICANTLY REDUCE THE EXPRESSION OF NIPP1 (NUCLEAR INHIBITOR OF PROTEIN PHOSPHATASE 1) IN PODOCYTES IN VITRO AS WELL AS IN HUMAN AND MURINE DKD. NIPP1 WAS SHOWN BY OTHERS TO INTERACT WITH ENHANCER OF ZESTE HOMOLOG 2 (EZH2), WHICH CATALYZES THE REPRESSIVE METHYLATION OF H3K27ME3 ON HISTONE 3. THEREFORE, WE HYPOTHESIZED THAT AGES CAN DIRECTLY INDUCE EPIGENETIC CHANGES IN PODOCYTES. METHODS: WE ANALYZED THE RELEVANCE OF AGES ON EZH2 EXPRESSION AND ACTIVITY IN A MURINE PODOCYTE CELL LINE. CELLS WERE TREATED WITH 5 MG/ML GLYCATED BSA FOR 24 H. TO DETERMINE THE MEANING OF EZH2 SUPPRESSION, EZH2 ACTIVITY WAS INHIBITED BY INCUBATING THE CELLS WITH THE PHARMACOLOGICAL METHYLTRANSFERASE INHIBITOR 3-DEAZANEPLANOCIN A; EZH2 EXPRESSION WAS REPRESSED WITH SIRNA. MRNA EXPRESSION WAS ANALYZED WITH REAL-TIME PCR, AND PROTEIN EXPRESSION WITH WESTERN BLOT. EZH2 EXPRESSION AND LEVEL OF H3K27 TRIMETHYLATION IN PODOCYTES OF DIABETIC DB/DB MICE, A MOUSE MODEL FOR TYPE 2 DM, WERE ANALYZED USING IMMUNOFLUORESCENCE. RESULTS: OUR DATA DEMONSTRATED THAT AGES DECREASE EZH2 EXPRESSION IN PODOCYTES AND CONSEQUENTLY REDUCE H3K27ME3. THIS SUPPRESSION OF EZH2 MIMICKED THE AGE EFFECTS AND CAUSED AN UPREGULATED EXPRESSION OF PATHOLOGICAL FACTORS THAT CONTRIBUTE TO PODOCYTE INJURY IN DKD. IN ADDITION, ANALYSES OF DB/DB MICE SHOWED SIGNIFICANTLY REDUCED H3K27ME3 AND EZH2 EXPRESSION IN PODOCYTES. MOREOVER, THE SUPPRESSION OF NIPP1 AND EZH2 SHOWED SIMILAR EFFECTS REGARDING PODOCYTE INJURY. CONCLUSIONS: OUR STUDIES PROVIDE A NOVEL PATHWAY HOW AGES CONTRIBUTE TO PODOCYTE INJURY AND THE FORMATION OF THE SO-CALLED METABOLIC MEMORY IN DKD. 2020 12 669 40 BONE MARROW STROMAL CELL ANTIGEN-1 (CD157) REGULATED BY SPHINGOSINE KINASE 2 MEDIATES KIDNEY FIBROSIS. CHRONIC KIDNEY DISEASE IS A PROGRESSIVE DISEASE THAT MAY LEAD TO END-STAGE RENAL DISEASE. INTERSTITIAL FIBROSIS DEVELOPS AS THE DISEASE PROGRESSES. THERAPIES THAT FOCUS ON FIBROSIS TO DELAY OR REVERSE PROGRESSIVE RENAL FAILURE ARE LIMITED. WE AND OTHERS SHOWED THAT SPHINGOSINE KINASE 2-DEFICIENT MICE (SPHK2 (-/-)) DEVELOP LESS FIBROSIS IN MOUSE MODELS OF KIDNEY FIBROSIS. SPHINGOSINE KINASE2 (SPHK2), ONE OF TWO SPHINGOSINE KINASES THAT PRODUCE SPHINGOSINE 1-PHOSPHATE (S1P), IS PRIMARILY LOCATED IN THE NUCLEUS. S1P PRODUCED BY SPHK2 INHIBITS HISTONE DEACETYLASE (HDAC) AND CHANGES HISTONE ACETYLATION STATUS, WHICH CAN LEAD TO ALTERED TARGET GENE EXPRESSION. WE HYPOTHESIZED THAT SPHK2 EPIGENETICALLY REGULATES DOWNSTREAM GENES TO INDUCE FIBROSIS, AND WE PERFORMED A COMPREHENSIVE ANALYSIS USING THE COMBINATION OF RNA-SEQ AND CHIP-SEQ. BST1/CD157 WAS IDENTIFIED AS A GENE THAT IS REGULATED BY SPHK2 THROUGH A CHANGE IN HISTONE ACETYLATION LEVEL, AND BST1 (-/-) MICE WERE FOUND TO DEVELOP LESS RENAL FIBROSIS AFTER UNILATERAL ISCHEMIA-REPERFUSION INJURY, A MOUSE MODEL OF KIDNEY FIBROSIS. ALTHOUGH BST1 IS A CELL-SURFACE MOLECULE THAT HAS A WIDE VARIETY OF FUNCTIONS THROUGH ITS VARIED ENZYMATIC ACTIVITIES AND DOWNSTREAM INTRACELLULAR SIGNALING PATHWAYS, NO STUDIES ON THE ROLE OF BST1 IN KIDNEY DISEASES HAVE BEEN REPORTED PREVIOUSLY. IN THE CURRENT STUDY, WE DEMONSTRATED THAT BST1 IS A GENE THAT IS REGULATED BY SPHK2 THROUGH EPIGENETIC CHANGE AND IS CRITICAL IN KIDNEY FIBROSIS. 2022 13 2779 42 EZH2 AND MATRIX CO-REGULATE PHENOTYPE AND KCNB2 EXPRESSION IN BLADDER SMOOTH MUSCLE CELLS. BACKGROUND: PARTIAL BLADDER OUTLET OBSTRUCTION (PBO) IS A WIDESPREAD CAUSE OF URINARY DYSFUNCTION AND PATIENT DISCOMFORT, RESULTING IN IMMENSE HEALTH CARE COSTS. PREVIOUSLY, WE FOUND THAT OBSTRUCTION IS ASSOCIATED WITH ALTERED REGULATION OF EPIGENETIC MACHINERY AND ALTERED FUNCTION. HERE WE EXAMINED IF PBO AND CHRONIC BLADDER OBSTRUCTIVE DISEASE (COBD) AFFECT EPIGENETIC MARKS IN A PROOF OF PRINCIPLE GENE AND EXPLORED MECHANISMS OF ITS EPIGENETIC REGULATION USING IN VITRO MODELS. METHODS: ARCHIVAL OBSTRUCTION TISSUES FROM COBD HAD BEEN CREATED IN 200-250 G FEMALE SPRAGUE-DAWLEY RATS BY SURGICAL LIGATION OF THE URETHRA FOR 6 WEEKS, FOLLOWED BY REMOVAL OF THE SUTURE AND FOLLOWING ANIMALS FOR 6 MORE WEEKS. OBSTRUCTION (PBO) IS THE 6-WEEK LIGATION ONLY. SHAM LIGATIONS COMPRISE PASSING THE SUTURE BEHIND THE URETHRA. HISTONE3 LYSINE27 TRIMETHYLATION (H3K27ME3) WAS STUDIED BY IMMUNOSTAINING AND CHROMATIN IMMUNOPRECIPITATION (CHIP)/PCR. THE INTERACTION OF MATRIX WITH KCNB2 REGULATION WAS STUDIED IN HUMAN BLADDER SMC PLATED ON DAMAGED MATRIX AND NATIVE COLLAGEN AND TREATED WITH VEHICLE OR UNC1999. CELLS WERE ANALYZED BY IMMUNOSTAINING FOR CELL PHENOTYPE, AND WESTERN BLOTTING FOR KCNB2, H3K27ME3 AND EZH2. EFFECTS OF CONDITIONED MEDIA FROM THESE CELLS WERE ALSO EXAMINED ON CELL PHENOTYPE. SIRNA AGAINST KCNB2 WAS EXAMINED FOR EFFECTS ON CELL PHENOTYPE AND GENE EXPRESSION BY RT-QPCR. RESULTS: H3K27ME3 INCREASED BY IMMUNOFLUORESCENCE DURING PBO, AND BY CHIP/PCR DURING COBD IN THE CPG ISLAND (CGI) AS WELL AS 350 BP UPSTREAM. OBSTRUCTION VS. SHAM ALSO SHOWED AN INCREASE IN H3K27ME3 DEPOSITION. IN SMC IN VITRO, EZH2 INHIBITION RESTORED KCNB2 EXPRESSION AND PARTIALLY RESTORED SMC PHENOTYPE. CONCLUSIONS: REGULATION OF KCNB2 AT THE PROMOTER DEMONSTRATED DYNAMIC CHANGES IN H3K27ME3 DURING COBD AND OBSTRUCTION. IN VITRO MODELS SUGGEST THAT MATRIX PLAYS A ROLE IN REGULATION OF EZH2, H3K27ME3 AND KCNB2, WHICH MAY PLAY A ROLE IN THE REGULATION OF SMOOTH MUSCLE PHENOTYPE IN VIVO. 2023 14 3096 37 GENOMIC CHARACTERIZATION REVEALS NOVEL MECHANISMS UNDERLYING THE VALOSIN-CONTAINING PROTEIN-MEDIATED CARDIAC PROTECTION AGAINST HEART FAILURE. CHRONIC HYPERTENSION IS A KEY RISK FACTOR FOR HEART FAILURE. HOWEVER, THE UNDERLYING MOLECULAR MECHANISMS ARE NOT FULLY UNDERSTOOD. OUR PREVIOUS STUDIES FOUND THAT THE VALOSIN-CONTAINING PROTEIN (VCP), AN ATPASE-ASSOCIATED PROTEIN, WAS SIGNIFICANTLY DECREASED IN THE HYPERTENSIVE HEART TISSUES. IN THIS STUDY, WE TESTED THE HYPOTHESIS THAT RESTORATION OF VCP PROTECTED THE HEART AGAINST PRESSURE OVERLOAD-INDUCED HEART FAILURE. WITH A CARDIAC-SPECIFIC TRANSGENIC (TG) MOUSE MODEL, WE SHOWED THAT A MODERATE INCREASE OF VCP WAS ABLE TO ATTENUATE CHRONIC PRESSURE OVERLOAD-INDUCED MALADAPTIVE CARDIAC HYPERTROPHY AND DYSFUNCTION. RNA SEQUENCING AND A COMPREHENSIVE BIOINFORMATIC ANALYSIS FURTHER DEMONSTRATED THAT OVEREXPRESSION OF VCP IN THE HEART NORMALIZED THE PRESSURE OVERLOAD-STIMULATED HYPERTROPHIC SIGNALS AND REPRESSED THE STRESS-INDUCED INFLAMMATORY RESPONSE. IN ADDITION, VCP OVEREXPRESSION PROMOTED CELL SURVIVAL BY ENHANCING THE MITOCHONDRIA RESISTANCE TO THE OXIDATIVE STRESS VIA ACTIVATING THE RICTOR-MEDIATED-GENE NETWORKS. VCP WAS ALSO FOUND TO BE INVOLVED IN THE REGULATION OF THE ALTERNATIVE SPLICING AND DIFFERENTIAL ISOFORM EXPRESSION FOR SOME GENES THAT ARE RELATED TO ATP PRODUCTION AND PROTEIN SYNTHESIS BY INTERACTING WITH LONG NO-CODING RNAS AND HISTONE DEACETYLASES, INDICATING A NOVEL EPIGENETIC REGULATION OF VCP IN INTEGRATING CODING AND NONCODING GENOMIC NETWORK IN THE STRESSED HEART. IN SUMMARY, OUR STUDY DEMONSTRATED THAT THE RESCUING OF A DEFICIENT VCP IN THE HEART COULD PREVENT PRESSURE OVERLOAD-INDUCED HEART FAILURE BY RECTIFYING CARDIAC HYPERTROPHIC AND INFLAMMATORY SIGNALING AND ENHANCING THE CARDIAC RESISTANCE TO OXIDATIVE STRESS, WHICH BROUGHT IN NOVEL INSIGHTS INTO THE UNDERSTANDING OF THE MECHANISM OF VCP IN PROTECTING PATIENTS FROM HYPERTENSIVE HEART FAILURE. 2020 15 4146 24 MECHANISMS REGULATING MUSCLE PROTEIN SYNTHESIS IN CKD. BACKGROUND: CKD INDUCES LOSS OF MUSCLE PROTEINS PARTLY BY SUPPRESSING MUSCLE PROTEIN SYNTHESIS. MUSCLES OF MICE WITH CKD HAVE INCREASED EXPRESSION OF NUCLEOLAR PROTEIN 66 (NO66), AS DO MUSCLE BIOPSY SPECIMENS FROM PATIENTS WITH CKD OR THOSE UNDERGOING HEMODIALYSIS. INFLAMMATION STIMULATES NO66 EXPRESSION AND CHANGES IN NF-KAPPAB MEDIATE THE RESPONSE. METHODS: SUBTOTAL NEPHRECTOMY CREATED A MOUSE MODEL OF CKD WITH BUN >80 MG/DL. CROSSING NO66(FLOX/FLOX) WITH MCK-CRE MICE BRED MUSCLE-SPECIFIC NO66 (MCK-NO66) KNOCKOUT MICE. EXPERIMENTS ASSESSED THE EFFECT OF REMOVING NO66. RESULTS: MUSCLE-SPECIFIC NO66 KNOCKOUT IN MICE BLOCKS CKD-INDUCED LOSS OF MUSCLE MASS AND IMPROVES PROTEIN SYNTHESIS. NO66 SUPPRESSION OF RIBOSOMAL BIOGENESIS VIA DEMETHYLASE ACTIVITY IS THE MECHANISM BEHIND THESE RESPONSES. IN MUSCLE CELLS, EXPRESSION OF NO66, BUT NOT OF DEMETHYLASE-DEAD MUTANT NO66, DECREASED H3K4ME3 AND H3K36ME3 AND SUPPRESSED PRE-RRNA EXPRESSION. KNOCKING OUT NO66 INCREASED THE ENRICHMENT OF H3K4ME3 AND H3K36ME3 ON RIBOSOMAL DNA. IN PRIMARY MUSCLE CELLS AND IN MUSCLES OF MICE WITHOUT NO66, RIBOSOMAL RNA, PRE-RRNA, AND PROTEIN SYNTHESIS ALL INCREASED. CONCLUSIONS: CKD SUPPRESSES MUSCLE PROTEIN SYNTHESIS VIA EPIGENETIC MECHANISMS THAT NO66 MEDIATES. BLOCKING NO66 COULD SUGGEST STRATEGIES THAT COUNTER CKD-INDUCED ABNORMAL MUSCLE PROTEIN CATABOLISM. 2020 16 4345 18 MIR-103 PROMOTES ENDOTHELIAL MALADAPTATION BY TARGETING LNCWDR59. BLOOD FLOW AT ARTERIAL BIFURCATIONS AND CURVATURES IS NATURALLY DISTURBED. ENDOTHELIAL CELLS (ECS) FAIL TO ADAPT TO DISTURBED FLOW, WHICH TRANSCRIPTIONALLY DIRECT ECS TOWARD A MALADAPTED PHENOTYPE, CHARACTERIZED BY CHRONIC REGENERATION OF INJURED ECS. MICRORNAS (MIRNAS) CAN REGULATE EC MALADAPTATION THROUGH TARGETING OF PROTEIN-CODING RNAS. HOWEVER, LONG NONCODING RNAS (LNCRNAS), KNOWN EPIGENETIC REGULATORS OF BIOLOGICAL PROCESSES, CAN ALSO BE MIRNA TARGETS, BUT THEIR CONTRIBUTION ON EC MALADAPTATION IS UNCLEAR. HERE WE SHOW THAT HYPERLIPIDEMIA- AND OXLDL-INDUCED UPREGULATION OF MIR-103 INHIBITS EC PROLIFERATION AND PROMOTES ENDOTHELIAL DNA DAMAGE THROUGH TARGETING OF NOVEL LNCWDR59. MIR-103 IMPEDES LNCWDR59 INTERACTION WITH NOTCH1-INHIBITOR NUMB, THEREFORE AFFECTING NOTCH1-INDUCED EC PROLIFERATION. MOREOVER, MIR-103 INCREASES THE SUSCEPTIBILITY OF PROLIFERATING ECS TO OXLDL-INDUCED MITOTIC ABERRATIONS, CHARACTERIZED BY AN INCREASED MICRONUCLEIC FORMATION AND DNA DAMAGE ACCUMULATION, BY AFFECTING NOTCH1-RELATED BETA-CATENIN CO-ACTIVATION. COLLECTIVELY, THESE DATA INDICATE THAT MIR-103 PROGRAMS ECS TOWARD A MALADAPTED PHENOTYPE THROUGH TARGETING OF LNCWDR59, WHICH MAY PROMOTE ATHEROSCLEROSIS. 2018 17 3962 30 LONG NONCODING RNA LEENE PROMOTES ANGIOGENESIS AND ISCHEMIC RECOVERY IN DIABETES MODELS. IMPAIRED ANGIOGENESIS IN DIABETES IS A KEY PROCESS CONTRIBUTING TO ISCHEMIC DISEASES SUCH AS PERIPHERAL ARTERIAL DISEASE. EPIGENETIC MECHANISMS, INCLUDING THOSE MEDIATED BY LONG NONCODING RNAS (LNCRNAS), ARE CRUCIAL LINKS CONNECTING DIABETES AND THE RELATED CHRONIC TISSUE ISCHEMIA. HERE WE IDENTIFY THE LNCRNA THAT ENHANCES ENDOTHELIAL NITRIC OXIDE SYNTHASE (ENOS) EXPRESSION (LEENE) AS A REGULATOR OF ANGIOGENESIS AND ISCHEMIC RESPONSE. LEENE EXPRESSION WAS DECREASED IN DIABETIC CONDITIONS IN CULTURED ENDOTHELIAL CELLS (ECS), MOUSE HIND LIMB MUSCLES, AND HUMAN ARTERIES. INHIBITION OF LEENE IN HUMAN MICROVASCULAR ECS REDUCED THEIR ANGIOGENIC CAPACITY WITH A DYSREGULATED ANGIOGENIC GENE PROGRAM. DIABETIC MICE DEFICIENT IN LEENE DEMONSTRATED IMPAIRED ANGIOGENESIS AND PERFUSION FOLLOWING HIND LIMB ISCHEMIA. IMPORTANTLY, OVEREXPRESSION OF HUMAN LEENE RESCUED THE IMPAIRED ISCHEMIC RESPONSE IN LEENE-KNOCKOUT MICE AT TISSUE FUNCTIONAL AND SINGLE-CELL TRANSCRIPTOMIC LEVELS. MECHANISTICALLY, LEENE RNA PROMOTED TRANSCRIPTION OF PROANGIOGENIC GENES IN ECS, SUCH AS KDR (ENCODING VEGFR2) AND NOS3 (ENCODING ENOS), POTENTIALLY BY INTERACTING WITH LEO1, A KEY COMPONENT OF THE RNA POLYMERASE II-ASSOCIATED FACTOR COMPLEX AND MYC, A CRUCIAL TRANSCRIPTION FACTOR FOR ANGIOGENESIS. TAKEN TOGETHER, OUR FINDINGS DEMONSTRATE AN ESSENTIAL ROLE FOR LEENE IN THE REGULATION OF ANGIOGENESIS AND TISSUE PERFUSION. FUNCTIONAL ENHANCEMENT OF LEENE TO RESTORE ANGIOGENESIS FOR TISSUE REPAIR AND REGENERATION MAY REPRESENT A POTENTIAL STRATEGY TO TACKLE ISCHEMIC VASCULAR DISEASES. 2023 18 5442 26 RENIN-ANGIOTENSIN BLOCKADE RESETS PODOCYTE EPIGENOME THROUGH KRUPPEL-LIKE FACTOR 4 AND ATTENUATES PROTEINURIA. PROTEINURIA IS A CENTRAL COMPONENT OF CHRONIC KIDNEY DISEASE AND AN INDEPENDENT RISK FACTOR FOR CARDIOVASCULAR DISEASE. KIDNEY PODOCYTES HAVE AN ESSENTIAL ROLE AS A FILTRATION BARRIER AGAINST PROTEINURIA. KRUPPEL-LIKE FACTOR 4 (KLF4) IS EXPRESSED IN PODOCYTES AND DECREASED IN GLOMERULAR DISEASES LEADING TO METHYLATION OF THE NEPHRIN PROMOTER, DECREASED NEPHRIN EXPRESSION AND PROTEINURIA. TREATMENT WITH AN ANGIOTENSIN RECEPTOR BLOCKER (ARB) REDUCED METHYLATION OF THE NEPHRIN PROMOTER IN MURINE GLOMERULI OF AN ADRIAMYCIN NEPHROPATHY MODEL WITH RECOVERY OF KLF4 EXPRESSION AND A DECREASE IN ALBUMINURIA. IN PODOCYTE-SPECIFIC KLF4 KNOCKOUT MICE, THE EFFECT OF ARB ON ALBUMINURIA AND THE NEPHRIN PROMOTER METHYLATION WAS ATTENUATED. IN CULTURED HUMAN PODOCYTES, ANGIOTENSIN II REDUCED KLF4 EXPRESSION AND CAUSED METHYLATION OF THE NEPHRIN PROMOTER WITH DECREASED NEPHRIN EXPRESSION. IN PATIENTS, NEPHRIN PROMOTER METHYLATION WAS INCREASED IN PROTEINURIC KIDNEY DISEASES WITH DECREASED KLF4 AND NEPHRIN EXPRESSION. KLF4 EXPRESSION IN ARB-TREATED PATIENTS WAS HIGHER IN PATIENTS WITH THAN WITHOUT ARB TREATMENT. THUS, ANGIOTENSIN II CAN MODULATE EPIGENETIC REGULATION IN PODOCYTES AND ARB INHIBITS THESE ACTIONS IN PART VIA KLF4 IN PROTEINURIC KIDNEY DISEASES. THIS STUDY PROVIDES A NEW CONCEPT THAT RENIN-ANGIOTENSIN SYSTEM BLOCKADE CAN EXERT THERAPEUTIC EFFECTS THROUGH EPIGENETIC MODULATION OF THE KIDNEY GENE EXPRESSION. 2015 19 2204 39 EPIGENETIC MODIFICATION OF THE VON WILLEBRAND FACTOR PROMOTER DRIVES PLATELET AGGREGATION ON THE PULMONARY ENDOTHELIUM IN CHRONIC THROMBOEMBOLIC PULMONARY HYPERTENSION. RATIONALE: VON WILLEBRAND FACTOR (VWF) MEDIATES PLATELET ADHESION DURING THROMBOSIS. WHILE CHRONIC THROMBOEMBOLIC PULMONARY HYPERTENSION (CTEPH) IS ASSOCIATED WITH INCREASED PLASMA LEVELS OF VWF, THE ROLE OF THIS PROTEIN IN CTEPH HAS REMAINED ENIGMATIC. OBJECTIVES: TO IDENTIFY THE ROLE OF VWF IN CTEPH. METHODS: CTEPH-SPECIFIC PATIENT PLASMA AND PULMONARY ENDARTERECTOMY MATERIAL FROM PATIENTS WITH CTEPH WERE USED TO STUDY THE RELATIONSHIP BETWEEN INFLAMMATION, VWF EXPRESSION, AND PULMONARY THROMBOSIS. CELL CULTURE FINDINGS WERE VALIDATED IN HUMAN TISSUE, AND PROTEOMICS AND CHROMATIN IMMUNOPRECIPITATION WERE USED TO INVESTIGATE THE UNDERLYING MECHANISM OF CTEPH. MEASUREMENTS AND MAIN RESULTS: VWF IS INCREASED IN PLASMA AND THE PULMONARY ENDOTHELIUM OF CTEPH PATIENTS. IN VITRO, THE INCREASE IN VWF GENE EXPRESSION AND THE HIGHER RELEASE OF VWF PROTEIN UPON ENDOTHELIAL ACTIVATION RESULTED IN ELEVATED PLATELET ADHESION TO CTEPH ENDOTHELIUM. PROTEOMIC ANALYSIS REVEALED THAT NUCLEAR FACTOR (NF)-KAPPAB2 WAS SIGNIFICANTLY INCREASED IN CTEPH. WE DEMONSTRATE REDUCED HISTONE TRI-METHYLATION AND INCREASED HISTONE ACETYLATION OF THE VWF PROMOTER IN CTEPH ENDOTHELIUM, FACILITATING BINDING OF NF-KAPPAB2 TO THE VWF PROMOTER AND DRIVING VWF TRANSCRIPTION. GENETIC INTERFERENCE OF NFKAPPAB2 NORMALIZED THE HIGH VWF RNA EXPRESSION LEVELS AND REVERSED THE PROTHROMBOTIC PHENOTYPE OBSERVED IN CTEPH-PULMONARY ARTERY ENDOTHELIAL CELLS. CONCLUSIONS: EPIGENETIC REGULATION OF THE VWF PROMOTER CONTRIBUTES TO THE CREATION OF A LOCAL ENVIRONMENT THAT FAVORS IN SITU THROMBOSIS IN THE PULMONARY ARTERIES. IT REVEALS A DIRECT MOLECULAR LINK BETWEEN INFLAMMATORY PATHWAYS AND PLATELET ADHESION IN THE PULMONARY VASCULAR WALL, EMPHASIZING A POSSIBLE ROLE OF IN SITU THROMBOSIS IN THE DEVELOPMENT OR PROGRESSION OF CTEPH. 2022 20 2041 35 EPIGENETIC CHANGES WITHIN THE ANNULUS FIBROSUS BY DNA METHYLATION IN RAT INTERVERTEBRAL DISC DEGENERATION MODEL. INTERVERTEBRAL DISC DEGENERATION (IDD) IS AN AGE-DEPENDENT PROGRESSIVE SPINAL DISEASE THAT CAUSES CHRONIC BACK OR NECK PAIN. ALTHOUGH AGING HAS LONG BEEN PRESENTED AS THE MAIN RISK FACTOR, THE EXACT CAUSE IS NOT FULLY KNOWN. DNA METHYLATION IS ASSOCIATED WITH CHRONIC PAIN, SUGGESTING THAT EPIGENETIC MODULATION MAY AMELIORATE DISC DEGENERATION. WE EXAMINED HISTOLOGICAL CHANGES IN THE DNA METHYLATION WITHIN THE DISCS AND THEIR ASSOCIATION WITH PAIN-RELATED TRANSIENT RECEPTOR POTENTIAL VANILLOID SUBTYPE 1 (TRPV1) EXPRESSION IN RATS SUBJECTED TO IDD. EPIGENETIC MARKERS (5-HYDROXYMETHYLCYTOSINE (5HMC), 5-METHYLCYTOSINE (5MC)), DNA METHYLTRANSFERASES (DNMTS), AND TEN-ELEVEN TRANSLOCATIONS (TETS) WERE ANALYZED USING IMMUNOHISTOCHEMISTRY, REAL-TIME PCR, AND DNA DOT-BLOT FOLLOWING IDD. RESULTS REVEALED HIGH 5MC LEVELS IN THE ANNULUS FIBROSUS (AF) REGION WITHIN THE DISC AFTER IDD AND AN ASSOCIATION WITH TRPV1 EXPRESSION. DNMT1 IS MAINLY INVOLVED IN 5MC CONVERSION IN DEGENERATED DISCS. HOWEVER, 5HMC LEVELS DID NOT DIFFER BETWEEN GROUPS. A DEGENERATED DISC CAN LEAD TO LOCOMOTOR DEFECTS AS ASSESSED BY LADDER AND TAIL SUSPENSION TESTS, NO PAIN SIGNALS IN THE VON FREY TEST, UPREGULATED MATRIX METALLOPROTEINASE-3, AND DOWNREGULATED AGGRECAN LEVELS WITHIN THE DISC. THUS, WE FOUND THAT THE DNA METHYLATION STATUS IN THE AF REGION OF THE DISC WAS MAINLY CHANGED AFTER IDD AND ASSOCIATED WITH ABERRANT TRPV1 EXPRESSION IN DEGENERATED DISCS. 2022