1 3124 135 GHRELIN TREATMENT IMPROVES PHYSICAL DECLINE IN SARCOPENIA MODEL MICE THROUGH MUSCULAR ENHANCEMENT AND MITOCHONDRIAL ACTIVATION. CHRONIC KIDNEY DISEASE (CKD) IMPAIRS PHYSICAL PERFORMANCE IN HUMANS, WHICH LEADS TO A RISK OF ALL-CAUSE MORTALITY. IN OUR PREVIOUS STUDY, WE DEMONSTRATED THAT A REDUCTION IN MUSCLE MITOCHONDRIA RATHER THAN MUSCLE MASS WAS A MAJOR CAUSE OF PHYSICAL DECLINE IN 5/6 NEPHRECTOMIZED CKD MODEL MICE. BECAUSE GHRELIN ADMINISTRATION HAS BEEN REPORTED TO ENHANCE OXYGEN UTILIZATION IN SKELETAL MUSCLE, WE EXAMINED THE USEFULNESS OF GHRELIN FOR A RECOVERY OF PHYSICAL DECLINE IN 5/6 NEPHRECTOMIZED C57BL/6 MICE, FOCUSING ON THE EPIGENETIC MODIFICATION OF PEROXISOME PROLIFERATOR ACTIVATED RECEPTOR GAMMA COACTIVATOR-1ALPHA (PGC-1ALPHA), A MASTER REGULATOR OF MITOCHONDRIAL BIOGENESIS. THE MICE WERE INTRAPERITONEALLY ADMINISTERED ACYLATED GHRELIN (0.1 NMOL/GBW; THREE TIMES PER WEEK) FOR A MONTH. MUSCLE STRENGTH AND EXERCISE ENDURANCE WERE MEASURED BY USING A DYNAMOMETER AND TREADMILL, RESPECTIVELY. MITOCHONDRIAL DNA COPY NUMBER WAS DETERMINED BY QUANTITATIVE PCR. THE METHYLATION LEVELS OF THE CYTOSINE RESIDUE AT 260 BASE PAIRS UPSTREAM OF THE TRANSLATION INITIATION POINT (C-260) OF PGC-1ALPHA, WHICH HAS BEEN DEMONSTRATED TO DECREASE THE EXPRESSION, WAS EVALUATED BY METHYLATION-SPECIFIC PCR AND BISULFITE GENOMIC SEQUENCING METHODS AFTER THE GHRELIN ADMINISTRATION. GHRELIN ADMINISTRATION IMPROVED BOTH MUSCLE STRENGTH AND EXERCISE ENDURANCE IN THE MICE AND WAS ASSOCIATED WITH AN INCREASE IN MUSCLE MASS AND MUSCLE MITOCHONDRIAL CONTENT. GHRELIN ADMINISTRATION DECREASED THE METHYLATION RATIO OF C-260 OF PGC-1ALPHA IN THE SKELETAL MUSCLE AND INCREASED THE EXPRESSION. THEREFORE, GHRELIN ADMINISTRATION EFFECTIVELY REDUCED THE PHYSICAL DECLINE IN 5/6 NEPHRECTOMIZED MICE AND WAS ACCOMPANIED WITH AN INCREASED MITOCHONDRIAL CONTENT THROUGH DE-METHYLATION OF THE PROMOTER REGION OF PGC-1ALPHA IN THE MUSCLE. 2017 2 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 3 1298 32 DECREASED NUCLEAR RECEPTOR ACTIVITY AND EPIGENETIC MODULATION ASSOCIATES WITH DOWN-REGULATION OF HEPATIC DRUG-METABOLIZING ENZYMES IN CHRONIC KIDNEY DISEASE. PATIENTS WITH CHRONIC KIDNEY DISEASE (CKD) REQUIRE MANY MEDICATIONS. CYP2C AND CYP3A DRUG-METABOLIZING ENZYMES PLAY A CRITICAL ROLE IN DETERMINING THE PHARMACOKINETICS OF THE MAJORITY OF PRESCRIBED MEDICATIONS. THESE ENZYMES ARE TRANSCRIPTIONALLY REGULATED BY THE NUCLEAR RECEPTORS PREGNANE X RECEPTOR (PXR) AND HEPATIC NUCLEAR FACTOR 4ALPHA (HNF-4ALPHA). EXPRESSION OF CYP2C AND CYP3A IS DECREASED IN CKD; HOWEVER, THE MECHANISMS BY WHICH THIS OCCURS IS UNKNOWN. WE INDUCED CKD IN RATS BY 5/6 NEPHRECTOMY AND USED CHROMATIN IMMUNOPRECIPITATION (CHIP) TO DETERMINE NUCLEAR RECEPTOR- AND EPIGENETIC ALTERATION-MEDIATED DIFFERENCES IN THE PROMOTER REGION OF THE CYP2C AND CYP3A GENES. RNA POLYMERASE II AND HNF-4ALPHA BINDING WAS DECREASED 76 AND 57% IN THE CYP2C11 PROMOTOR AND 71 AND 77% IN THE CYP3A2 PROMOTER, RESPECTIVELY (P<0.05). CHIP ALSO REVEALED A 57% DECREASE IN PXR BINDING TO THE CYP3A2 PROMOTER IN CKD RATS (P<0.05). THE DECREASE IN PXR AND HNF-4ALPHA BINDING WAS ACCOMPANIED BY DIMINISHED HISTONE 4 ACETYLATION IN THE CYP3A2 PROMOTER (48%) AND HISTONE 3 ACETYLATION IN THE CYP2C11 (77%) AND CYP3A2 (77%) PROMOTER LOCI FOR NUCLEAR RECEPTOR ACTIVATION (P<0.05). THIS STUDY SUGGESTS THAT DECREASED NUCLEAR RECEPTOR BINDING AND HISTONE ACETYLATION MAY CONTRIBUTE TO THE MECHANISM OF DRUG-METABOLIZING ENZYME DOWN-REGULATION AND ALTERED PHARMACOKINETICS IN CKD. 2014 4 5675 29 SHIFTS IN PODOCYTE HISTONE H3K27ME3 REGULATE MOUSE AND HUMAN GLOMERULAR DISEASE. HISTONE PROTEIN MODIFICATIONS CONTROL FATE DETERMINATION DURING NORMAL DEVELOPMENT AND DEDIFFERENTIATION DURING DISEASE. HERE, WE SET OUT TO DETERMINE THE EXTENT TO WHICH DYNAMIC CHANGES TO HISTONES AFFECT THE DIFFERENTIATED PHENOTYPE OF ORDINARILY QUIESCENT ADULT GLOMERULAR PODOCYTES. TO DO THIS, WE EXAMINED THE CONSEQUENCES OF SHIFTING THE BALANCE OF THE REPRESSIVE HISTONE H3 LYSINE 27 TRIMETHYLATION (H3K27ME3) MARK IN PODOCYTES. ADRIAMYCIN NEPHROTOXICITY AND SUBTOTAL NEPHRECTOMY (SNX) STUDIES INDICATED THAT DELETION OF THE HISTONE METHYLATING ENZYME EZH2 FROM PODOCYTES DECREASED H3K27ME3 LEVELS AND SENSITIZED MICE TO GLOMERULAR DISEASE. H3K27ME3 WAS ENRICHED AT THE PROMOTER REGION OF THE NOTCH LIGAND JAG1 IN PODOCYTES, AND DEREPRESSION OF JAG1 BY EZH2 INHIBITION OR KNOCKDOWN FACILITATED PODOCYTE DEDIFFERENTIATION. CONVERSELY, INHIBITION OF THE JUMONJI C DOMAIN-CONTAINING DEMETHYLASES JMJD3 AND UTX INCREASED THE H3K27ME3 CONTENT OF PODOCYTES AND ATTENUATED GLOMERULAR DISEASE IN ADRIAMYCIN NEPHROTOXICITY, SNX, AND DIABETES. PODOCYTES IN GLOMERULI FROM HUMANS WITH FOCAL SEGMENTAL GLOMERULOSCLEROSIS OR DIABETIC NEPHROPATHY EXHIBITED DIMINISHED H3K27ME3 AND HEIGHTENED UTX CONTENT. ANALOGOUS TO HUMAN DISEASE, INHIBITION OF JMJD3 AND UTX ABATED NEPHROPATHY PROGRESSION IN MICE WITH ESTABLISHED GLOMERULAR INJURY AND REDUCED H3K27ME3 LEVELS. TOGETHER, THESE FINDINGS INDICATE THAT OSTENSIBLY STABLE CHROMATIN MODIFICATIONS CAN BE DYNAMICALLY REGULATED IN QUIESCENT CELLS AND THAT EPIGENETIC REPROGRAMMING CAN IMPROVE OUTCOMES IN GLOMERULAR DISEASE BY REPRESSING THE REACTIVATION OF DEVELOPMENTAL PATHWAYS. 2018 5 3604 74 IMPROVEMENT OF PHYSICAL DECLINE THROUGH COMBINED EFFECTS OF MUSCLE ENHANCEMENT AND MITOCHONDRIAL ACTIVATION BY A GASTRIC HORMONE GHRELIN IN MALE 5/6NX CKD MODEL MICE. BECAUSE A PHYSICAL DECLINE CORRELATES WITH AN INCREASED RISK OF A WIDE RANGE OF DISEASE AND MORBIDITY, AN IMPROVEMENT OF PHYSICAL PERFORMANCE IS EXPECTED TO BRING SIGNIFICANT CLINICAL BENEFITS. THE PRIMARY CAUSE OF PHYSICAL DECLINE IN 5/6 NEPHRECTOMIZED (5/6NX) CHRONIC KIDNEY DISEASE MODEL MICE HAS BEEN REGARDED AS A DECREASE IN MUSCLE MASS; HOWEVER, OUR RECENT STUDY SHOWED THAT A DECREASE IN MUSCLE MITOCHONDRIA PLAYS A CRITICAL ROLE. IN THE PRESENT STUDY, WE EXAMINED THE EFFECTS OF A GASTRIC HORMONE GHRELIN, WHICH HAS BEEN REPORTED TO PROMOTE MUSCLE MITOCHONDRIAL OXIDATION, ON THE PHYSICAL DECLINE IN THE CHRONIC KIDNEY DISEASE MODEL MICE, FOCUSING ON THE EPIGENETIC MODULATIONS OF A MITOCHONDRIAL ACTIVATOR GENE, PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-GAMMA COACTIVATOR-1ALPHA (PGC-1ALPHA). GHRELIN TREATMENT IMPROVED A DECLINE IN EXERCISE ENDURANCE OF 5/6NX MICE, ASSOCIATED WITH AN INCREASE IN BOTH OF THE MUSCLE MASS AND MITOCHONDRIAL AMOUNT. THE EXPRESSION LEVEL OF PGC-1ALPHA WAS DECREASED IN THE SKELETAL MUSCLE OF 5/6NX MICE, WHICH WAS ASSOCIATED WITH AN INCREASE IN THE METHYLATION RATIO OF THE CYTOSINE RESIDUE AT 260 BASE PAIRS UPSTREAM OF THE INITIATION POINT. CONVERSELY, GHRELIN TREATMENT DE-METHYLATED THE CYTOSINE RESIDUE AND INCREASED THE EXPRESSION OF PGC-1ALPHA. A REPRESENTATIVE MUSCLE ANABOLIC FACTOR, IGF-1, DID NOT AFFECT THE EXPRESSION OF PGC-1ALPHA AND MUSCLE MITOCHONDRIAL AMOUNT, ALTHOUGH IT INCREASED MUSCLE MASS. AS A RESULT, IGF-1 TREATMENT IN 5/6NX MICE DID NOT INCREASE THE DECREASED EXERCISE ENDURANCE AS EFFECTIVELY AS GHRELIN TREATMENT DID. THESE FINDINGS INDICATE AN ADVANTAGE OF GHRELIN TREATMENT FOR A RECOVERY OF PHYSICAL DECLINE. 2015 6 6702 36 VEGFA PROMOTER GENE HYPERMETHYLATION AT HIF1ALPHA BINDING SITE IS AN EARLY CONTRIBUTOR TO CKD PROGRESSION AFTER RENAL ISCHEMIA. CHRONIC HYPOXIA IS A MAJOR CONTRIBUTOR TO CHRONIC KIDNEY DISEASE (CKD) AFTER ACUTE KIDNEY INJURY (AKI). HOWEVER, THE TEMPORAL RELATION BETWEEN THE ACUTE INSULT AND MALADAPTIVE RENAL RESPONSE TO HYPOXIA REMAINS UNCLEAR. IN THIS STUDY, WE ANALYZED THE TIME-COURSE OF RENAL HEMODYNAMICS, OXIDATIVE STRESS, INFLAMMATION, AND FIBROSIS, AS WELL AS EPIGENETIC MODIFICATIONS, WITH FOCUS ON HIF1ALPHA/VEGF SIGNALING, IN THE AKI TO CKD TRANSITION. SHAM-OPERATED, RIGHT NEPHRECTOMY (UNX), AND UNX PLUS RENAL ISCHEMIA (IR + UNX) GROUPS OF RATS WERE INCLUDED AND STUDIED AT 1, 2, 3, OR 4 MONTHS. THE IR + UNX GROUP DEVELOPED CKD CHARACTERIZED BY PROGRESSIVE PROTEINURIA, RENAL DYSFUNCTION, TUBULAR PROLIFERATION, AND FIBROSIS. AT FIRST MONTH POST-ISCHEMIA, THERE WAS A TWOFOLD SIGNIFICANT INCREASE IN OXIDATIVE STRESS AND REDUCTION IN GLOBAL DNA METHYLATION THAT WAS MAINTAINED THROUGHOUT THE STUDY. HIF1ALPHA AND VEGFA EXPRESSION WERE DEPRESSED IN THE FIRST AND SECOND-MONTHS POST-ISCHEMIA, AND THEN HIF1ALPHA BUT NOT VEGFA EXPRESSION WAS RECOVERED. INTERESTINGLY, HYPERMETHYLATION OF THE VEGFA PROMOTER GENE AT THE HIF1ALPHA BINDING SITE WAS FOUND, SINCE EARLY STAGES OF THE CKD PROGRESSION. OUR FINDINGS SUGGEST THAT RENAL HYPOPERFUSION, INEFFICIENT HYPOXIC RESPONSE, INCREASED OXIDATIVE STRESS, DNA HYPOMETHYLATION, AND, VEGFA PROMOTER GENE HYPERMETHYLATION AT HIF1ALPHA BINDING SITE, ARE EARLY DETERMINANTS OF AKI-TO-CKD TRANSITION. 2021 7 1274 28 DACH1 PROTECTS PODOCYTES FROM EXPERIMENTAL DIABETIC INJURY AND MODULATES PTIP-H3K4ME3 ACTIVITY. DACHSHUND HOMOLOG 1 (DACH1), A KEY CELL-FATE DETERMINANT, REGULATES TRANSCRIPTION BY DNA SEQUENCE-SPECIFIC BINDING. WE IDENTIFIED DIMINISHED DACH1 EXPRESSION IN A LARGE-SCALE SCREEN FOR MUTATIONS THAT CONVERT INJURY-RESISTANT PODOCYTES INTO INJURY-SUSCEPTIBLE PODOCYTES. IN DIABETIC KIDNEY DISEASE (DKD) PATIENTS, PODOCYTE DACH1 EXPRESSION LEVELS ARE DIMINISHED, A CONDITION THAT STRONGLY CORRELATES WITH POOR CLINICAL OUTCOMES. GLOBAL DACH1 KO MICE MANIFEST RENAL HYPOPLASIA AND DIE PERINATALLY. PODOCYTE-SPECIFIC DACH1 KO MICE, HOWEVER, MAINTAIN NORMAL GLOMERULAR ARCHITECTURE AT BASELINE, BUT RAPIDLY EXHIBIT PODOCYTE INJURY AFTER DIABETES ONSET. FURTHERMORE, PODOCYTE-SPECIFIC AUGMENTATION OF DACH1 EXPRESSION IN MICE PROTECTS FROM DKD. COMBINED RNA SEQUENCING AND IN SILICO PROMOTER ANALYSIS REVEAL CONVERSELY OVERLAPPING GLOMERULAR TRANSCRIPTOMIC SIGNATURES BETWEEN PODOCYTE-SPECIFIC DACH1 AND PAX TRANSACTIVATION-DOMAIN INTERACTING PROTEIN (PTIP) KO MICE, WITH UPREGULATED GENES POSSESSING HIGHER-THAN-EXPECTED NUMBERS OF PROMOTER DACH1-BINDING SITES. PTIP, AN ESSENTIAL COMPONENT OF THE ACTIVATING HISTONE H3 LYSINE 4 TRIMETHYLATION (H3K4ME3) COMPLEX, INTERACTS WITH DACH1 AND IS RECRUITED BY DACH1 TO ITS PROMOTER-BINDING SITES. DACH1-PTIP RECRUITMENT REPRESSES TRANSCRIPTION AND REDUCES PROMOTER H3K4ME3 LEVELS. DACH1 KNOCKDOWN IN PODOCYTES COMBINED WITH HYPERGLYCEMIA TRIGGERS TARGET GENE UPREGULATION AND INCREASES PROMOTER H3K4ME3. THESE FINDINGS REVEAL THAT IN DKD, DIMINISHED DACH1 EXPRESSION ENHANCES PODOCYTE INJURY VULNERABILITY VIA EPIGENETIC DEREPRESSION OF ITS TARGET GENES. 2021 8 4226 27 METHYLATION IN PERICYTES AFTER ACUTE INJURY PROMOTES CHRONIC KIDNEY DISEASE. THE ORIGIN AND FATE OF RENAL MYOFIBROBLASTS IS NOT CLEAR AFTER ACUTE KIDNEY INJURY (AKI). HERE, WE DEMONSTRATE THAT MYOFIBROBLASTS WERE ACTIVATED FROM QUIESCENT PERICYTES (QPERICYTES) AND THE CELL NUMBERS INCREASED AFTER ISCHEMIA/REPERFUSION INJURY-INDUCED AKI (IRI-AKI). MYOFIBROBLASTS UNDERWENT APOPTOSIS DURING RENAL RECOVERY BUT ONE-FIFTH OF THEM SURVIVED IN THE RECOVERED KIDNEYS ON DAY 28 AFTER IRI-AKI AND THEIR CELL NUMBERS INCREASED AGAIN AFTER DAY 56. MICROARRAY DATA SHOWED THE DISTINCTIVE GENE EXPRESSION PATTERNS OF QPERICYTES, ACTIVATED PERICYTES (APERICYTES, MYOFIBROBLASTS), AND INACTIVATED PERICYTES (IPERICYTES) ISOLATED FROM KIDNEYS BEFORE, ON DAY 7, AND ON DAY 28 AFTER IRI-AKI. HYPERMETHYLATION OF THE ACTA2 REPRESSOR YBX2 DURING IRI-AKI RESULTED IN EPIGENETIC MODIFICATION OF IPERICYTES TO PROMOTE THE TRANSITION TO CHRONIC KIDNEY DISEASE (CKD) AND AGGRAVATED FIBROGENESIS INDUCED BY A SECOND AKI INDUCED BY ADENINE. MECHANISTICALLY, TRANSFORMING GROWTH FACTOR-BETA1 DECREASED THE BINDING OF YBX2 TO THE PROMOTER OF ACTA2 AND INDUCED YBX2 HYPERMETHYLATION, THEREBY INCREASING ALPHA-SMOOTH MUSCLE ACTIN EXPRESSION IN APERICYTES. DEMETHYLATION BY 5-AZACYTIDINE RECOVERED THE MICROVASCULAR STABILIZING FUNCTION OF APERICYTES, REVERSED THE PROFIBROTIC PROPERTY OF IPERICYTES, PREVENTED AKI-CKD TRANSITION, AND ATTENUATED FIBROGENESIS INDUCED BY A SECOND ADENINE-AKI. IN CONCLUSION, INTERVENTION TO ERASE HYPERMETHYLATION OF PERICYTES AFTER AKI PROVIDES A STRATEGY TO STOP THE TRANSITION TO CKD. 2020 9 4356 33 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 10 5990 24 TGF-BETA1 PROMOTES EXPRESSION OF FIBROSIS-RELATED GENES THROUGH THE INDUCTION OF HISTONE VARIANT H3.3 AND HISTONE CHAPERONE HIRA. RENAL FIBROSIS IS A HISTOLOGICAL MANIFESTATION THAT OCCURS IN ALMOST EVERY TYPE OF CHRONIC KIDNEY DISEASE. HISTONE VARIANT H3.3 AND ITS CHAPERONE, HISTONE CELL CYCLE REGULATION DEFECTIVE HOMOLOG A (HIRA), SERVE AS EPIGENETIC MARKS THAT REGULATE TRANSCRIPTIONAL ACTIVITY. IN THIS STUDY, WE ASSESSED THE ROLES OF HISTONE H3.3 AND HIRA IN UNILATERAL URETERAL-OBSTRUCTION (UUO) MICE. IN UUO MICE, THE LEVELS OF HISTONE H3.3 AND HIRA WERE SIGNIFICANTLY UPREGULATED IN THE KIDNEYS. THESE UPREGULATED LEVELS WERE DECREASED BY A TGF-BETA1 NEUTRALIZING ANTIBODY. TGF-BETA1 INDUCED HISTONE H3.3 AND HIRA EXPRESSION IN VITRO VIA A SMAD3-DEPENDENT PATHWAY IN NORMAL RAT KIDNEY (NRK)-52E CELLS. ADDITIONALLY, KNOCKDOWN OF HIRA EXPRESSION DECREASED HISTONE H3.3 EXPRESSION AND FIBROGENESIS IN NRK-52E CELLS AFTER TGF-BETA1 STIMULATION. CHROMATIN IMMUNOPRECIPITATION ANALYSIS REVEALED THAT PROMOTERS OF FIBROSIS-RELATED GENES WERE IMMUNOPRECIPITATED WITH BOTH HISTONE H3.3 AND HIRA IN NRK-52E CELLS. LASTLY, IN HUMAN KIDNEY BIOPSIES FROM PATIENTS DIAGNOSED WITH IGA NEPHROPATHY, HISTONE H3.3 AND HIRA IMMUNOSTAINING CORRELATED POSITIVELY WITH AREAS OF FIBROSIS AND ESTIMATED GLOMERULAR FILTRATION RATE. IN CONCLUSION, TGF-BETA1 INDUCES EXPRESSION OF HISTONE H3.3 AND HIRA, WHICH REGULATES EXPRESSION OF FIBROSIS-RELATED GENES. 2018 11 5442 32 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 12 1015 24 CIGARETTE SMOKING CAUSES EPIGENETIC CHANGES ASSOCIATED WITH CARDIORENAL FIBROSIS. CLINICAL STUDIES INDICATE THAT SMOKING COMBUSTIBLE CIGARETTES PROMOTES PROGRESSION OF RENAL AND CARDIAC INJURY, LEADING TO FUNCTIONAL DECLINE IN THE SETTING OF CHRONIC KIDNEY DISEASE (CKD). HOWEVER, BASIC STUDIES USING IN VIVO SMALL ANIMAL MODELS THAT MIMIC CLINICAL PATHOLOGY OF CKD ARE LACKING. TO ADDRESS THIS ISSUE, WE EVALUATED RENAL AND CARDIAC INJURY PROGRESSION AND FUNCTIONAL CHANGES INDUCED BY 4 WK OF DAILY COMBUSTIBLE CIGARETTE SMOKE EXPOSURE IN THE 5/6TH PARTIAL NEPHRECTOMY (PNX) CKD MODEL. MOLECULAR EVALUATIONS REVEALED THAT CIGARETTE SMOKE SIGNIFICANTLY (P < 0.05) DECREASED RENAL AND CARDIAC EXPRESSION OF THE ANTIFIBROTIC MICRORNA MIR-29B-3 AND INCREASED EXPRESSION OF MOLECULAR FIBROSIS MARKERS. IN TERMS OF CARDIAC AND RENAL ORGAN STRUCTURE AND FUNCTION, EXPOSURE TO CIGARETTE SMOKE LED TO SIGNIFICANTLY INCREASED SYSTOLIC BLOOD PRESSURE, CARDIAC HYPERTROPHY, CARDIAC AND RENAL FIBROSIS, AND DECREASED RENAL FUNCTION. THESE DATA INDICATE THAT DECREASED EXPRESSION OF MIR-29B-3P IS A NOVEL MECHANISM WHEREIN CIGARETTE SMOKE PROMOTES ACCELERATED CARDIAC AND RENAL TISSUE INJURY IN CKD. (155 WORDS). 2016 13 6233 22 THE LONG NONCODING RNA TUG1 CONNECTS METABOLIC CHANGES WITH KIDNEY DISEASE IN PODOCYTES. AN INCREASING AMOUNT OF EVIDENCE SUGGESTS THAT METABOLIC ALTERATIONS PLAY A KEY ROLE IN CHRONIC KIDNEY DISEASE (CKD) PATHOGENESIS. IN THIS ISSUE OF THE JCI, LONG ET AL. REPORT THAT THE LONG NONCODING RNA (LNCRNA) TAURINE-UPREGULATED 1 (TUG1) CONTRIBUTES TO CKD DEVELOPMENT. THE AUTHORS SHOW THAT TUG1 REGULATES MITOCHONDRIAL FUNCTION IN PODOCYTES BY EPIGENETIC TARGETING OF EXPRESSION OF THE TRANSCRIPTION FACTOR PPARGAMMA COACTIVATOR 1ALPHA (PGC-1ALPHA, ENCODED BY PPARGC1A). TRANSGENIC OVEREXPRESSION OF TUG1 SPECIFICALLY IN PODOCYTES AMELIORATED DIABETES-INDUCED CKD IN MICE. TOGETHER, THESE RESULTS HIGHLIGHT AN IMPORTANT CONNECTION BETWEEN LNCRNA-MEDIATED METABOLIC ALTERATIONS IN PODOCYTES AND KIDNEY DISEASE DEVELOPMENT. 2016 14 3526 33 IL-6 AND SIL-6R INDUCES STAT3-DEPENDENT DIFFERENTIATION OF HUMAN VSMCS INTO OSTEOBLAST-LIKE CELLS THROUGH JMJD2B-MEDIATED HISTONE DEMETHYLATION OF RUNX2. INFLAMMATION AND VASCULAR CALCIFICATION ARE INDEPENDENT RISK FACTORS OF CARDIOVASCULAR EVENTS. VASCULAR SMOOTH MUSCLE CELLS (VSMCS) EXHIBIT OSTEOBLAST-LIKE CHARACTERISTICS IN RESPONSE TO VARIOUS STIMULI SUCH AS OXIDIZED CHOLESTEROL AND INFLAMMATION. HOWEVER THE PRECISE MECHANISM OF TRANSCRIPTIONAL REGULATION OF VSMCS BY INFLAMMATORY STIMULI REMAINS UNCLEAR. WE INVESTIGATED THE PROCESS AND MECHANISMS OF INFLAMMATORY CYTOKINE-INDUCED TRANSFORMATION OF HUMAN VSMCS (HVSMCS) INTO OSTEOBLAST-LIKE CELLS, WITH A SPECIAL FOCUS ON EPIGENETIC CHANGES. OUR RESULTS DEMONSTRATED: (1) INTERLEUKIN-6 (IL-6)/SOLUBLE INTERLEUKIN-6 RECEPTOR (SIL-6R) INDUCED TRANSFORMATION OF HVSMCS INTO AN OSTEOBLAST PHENOTYPE, WITH SUBSEQUENT VASCULAR CALCIFICATION, BASED ON THE RESULTS OF ALIZARIN RED S STAINING AND O-CRESOLPHTHALEIN COMPLEXONE METHOD; (2) IL-6/SIL-6R ACCELERATED THE EXPRESSION OF RUNT-RELATED TRANSCRIPTION FACTOR 2 (RUNX2) BASED ON THE RESULTS OF QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION; (3) KNOCKDOWN OF SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION (STAT) 3 REDUCED IL-6/SIL-6R-INDUCED RUNX2 MRNA EXPRESSION AND OSTEOBLAST TRANSDIFFERENTIATION OF HVSMCS; (4) CHROMATIN IMMUNOPRECIPITATION (CHIP) COUPLED WITH PCR (CHIP-PCR) IDENTIFIED A STAT-BINDING SITE IN RUNX2 PROMOTER REGION CONTAINING TRIMETHYLATED HISTONE 3 LYSINE 9 (H3K9ME3), A TRANSCRIPTIONAL REPRESSOR, AND H3K4ME3, A TRANSCRIPTIONAL ENHANCER. STIMULATION WITH IL-6/SIL-6R SUPPRESSED H3K9ME3 BUT NOT H3K4ME3 THROUGH THE RECRUITMENT OF JUMONJI DOMAIN-CONTAINING PROTEIN (JMJD) 2B, A HISTONE LYSINE DEMETHYLASE, AT THE STAT-BINDING SITE IN RUNX2 PROMOTER REGION; (5) IL-6/SIL-6R-INDUCED RUNX2 GENE EXPRESSION WAS INHIBITED IN HVSMCS PRETREATED WITH JIB04, JMJD2 INHIBITOR, AND THE INHIBITORY EFFECT WAS JIB04 DOSE-DEPENDENT. OUR RESULTS INDICATE THAT THE IL-6/STAT3/JMJD2B PATHWAY REGULATES HVSMCS DIFFERENTIATION INTO OSTEOBLAST-LIKE CELLS, WHICH SUGGEST ITS PATHOGENIC ROLE IN VASCULAR CALCIFICATION ASSOCIATED WITH CHRONIC INFLAMMATION. 2019 15 5332 27 PYRUVATE DEHYDROGENASE KINASE 1 AND 2 DEFICIENCY REDUCES HIGH-FAT DIET-INDUCED HYPERTROPHIC OBESITY AND INHIBITS THE DIFFERENTIATION OF PREADIPOCYTES INTO MATURE ADIPOCYTES. OBESITY IS NOW RECOGNIZED AS A DISEASE. THIS STUDY REVEALED A NOVEL ROLE FOR PYRUVATE DEHYDROGENASE KINASE (PDK) IN DIET-INDUCED HYPERTROPHIC OBESITY. MICE WITH GLOBAL OR ADIPOSE TISSUE-SPECIFIC PDK2 DEFICIENCY WERE PROTECTED AGAINST DIET-INDUCED OBESITY. THE WEIGHT OF ADIPOSE TISSUES AND THE SIZE OF ADIPOCYTES WERE REDUCED. ADIPOCYTE-SPECIFIC PDK2 DEFICIENCY SLIGHTLY INCREASED INSULIN SENSITIVITY IN HFD-FED MICE. IN STUDIES WITH 3T3-L1 PREADIPOCYTES, PDK2 AND PDK1 EXPRESSION WAS STRONGLY INCREASED DURING ADIPOGENESIS. EVIDENCE WAS FOUND FOR EPIGENETIC INDUCTION OF BOTH PDK1 AND PDK2. GAIN- AND LOSS-OF-FUNCTION STUDIES WITH 3T3-L1 CELLS REVEALED A CRITICAL ROLE FOR PDK1/2 IN ADIPOCYTE DIFFERENTIATION AND LIPID ACCUMULATION. PDK1/2 INDUCTION DURING DIFFERENTIATION WAS ALSO ACCOMPANIED BY INCREASED EXPRESSION OF HYPOXIA-INDUCIBLE FACTOR-1ALPHA (HIF1ALPHA) AND ENHANCED LACTATE PRODUCTION, BOTH OF WHICH WERE ABSENT IN THE CONTEXT OF PDK1/2 DEFICIENCY. EXOGENOUS LACTATE SUPPLEMENTATION INCREASED THE STABILITY OF HIF1ALPHA AND PROMOTED ADIPOGENESIS. PDK1/2 OVEREXPRESSION-MEDIATED ADIPOGENESIS WAS ABOLISHED BY HIF1ALPHA INHIBITION, SUGGESTING A ROLE FOR THE PDK-LACTATE-HIF1ALPHA AXIS DURING ADIPOGENESIS. IN HUMAN ADIPOSE TISSUE, THE EXPRESSION OF PDK1/2 WAS POSITIVELY CORRELATED WITH THAT OF THE ADIPOGENIC MARKER PPARGAMMA AND INVERSELY CORRELATED WITH OBESITY. SIMILARLY, PDK1/2 EXPRESSION IN MOUSE ADIPOSE TISSUE WAS DECREASED BY CHRONIC HIGH-FAT DIET FEEDING. WE CONCLUDE THAT PDK1 AND 2 ARE NOVEL REGULATORS OF ADIPOGENESIS THAT PLAY CRITICAL ROLES IN OBESITY. 2021 16 5504 29 RHEIN REVERSAL OF DNA HYPERMETHYLATION-ASSOCIATED KLOTHO SUPPRESSION AMELIORATES RENAL FIBROSIS IN MICE. RENAL FIBROSIS IS THE HALLMARK OF CHRONIC KIDNEY DISEASES (CKD) AND ITS DEVELOPMENT AND PROGRESSION ARE SIGNIFICANTLY AFFECTED BY EPIGENETIC MODIFICATIONS. RHEIN, A PLANT-DERIVED ANTHRAQUINONE, DISPLAYS STRONG ANTI-FIBROSIS PROPERTIES, BUT ITS PROTECTIVE MODE OF ACTION REMAINS INCOMPLETELY UNDERSTOOD. HERE WE EXPLORE THE MECHANISM OF RHEIN ANTI-RENAL FIBROSIS BY INVESTIGATING ITS REGULATION OF KLOTHO, A KNOWN RENAL ANTI-FIBROTIC PROTEIN WHOSE SUPPRESSION AFTER RENAL INJURY REPORTEDLY INVOLVES ABERRANT DNA METHYLATION. WE REPORT THAT RHEIN IS AN IMPRESSIVE UP-REGULATOR OF KLOTHO AND IT MARKEDLY REVERSED KLOTHO DOWN-REGULATION IN UNILATERAL URETERAL OCCLUSION-INDUCED FIBROTIC KIDNEY. FURTHER EXAMINATIONS REVEALED THAT KLOTHO LOSS IN FIBROTIC KIDNEY IS ASSOCIATED WITH KLOTHO PROMOTER HYPERMETHYLATION DUE TO ABERRANT METHYLTRANSFERASE 1 AND 3A EXPRESSIONS. HOWEVER, RHEIN SIGNIFICANTLY CORRECTED ALL THESE EPIGENETIC ALTERATIONS AND SUBSEQUENTLY ALLEVIATED PRO-FIBROTIC PROTEIN EXPRESSION AND RENAL FIBROSIS, WHEREAS KLOTHO KNOCKDOWN VIA RNA INTERFERENCES LARGELY ABROGATED THE ANTI-RENAL FIBROTIC EFFECTS OF RHEIN, SUGGESTING THAT RHEIN EPIGENETIC REVERSAL OF KLOTHO LOSS REPRESENTS A CRITICAL MODE OF ACTION THAT CONFERS RHEIN'S ANTI- RENAL FIBROTIC FUNCTIONS. ALTOGETHER OUR STUDIES UNCOVER A NOVEL HYPOMETHYLATING CHARACTER OF RHEIN IN PREVENTING KLOTHO LOSS AND RENAL FIBROSIS, AND DEMONSTRATE THE EFFICACY OF KLOTHO-TARGETED EPIGENETIC INTERVENTION IN POTENTIAL TREATMENT OF RENAL FIBROSIS-ASSOCIATED KIDNEY DISEASES. 2016 17 4001 41 LOSS OF MEN1 LEADS TO RENAL FIBROSIS AND DECREASES HGF-ADAMTS5 PATHWAY ACTIVITY VIA AN EPIGENETIC MECHANISM. BACKGROUND: RENAL FIBROSIS IS A SERIOUS CONDITION THAT RESULTS IN THE DEVELOPMENT OF CHRONIC KIDNEY DISEASES. THE MEN1 GENE IS AN EPIGENETIC REGULATOR THAT ENCODES THE MENIN PROTEIN AND ITS ROLE IN KIDNEY TISSUE REMAINS UNCLEAR. METHODS: KIDNEY HISTOLOGY WAS EXAMINED ON PARAFFIN SECTIONS STAINED WITH HEMATOXYLIN-EOSIN STAINING. MASSON'S TRICHROME STAINING AND SIRIUS RED STAINING WERE USED TO ANALYZE RENAL FIBROSIS. GENE AND PROTEIN EXPRESSION WERE DETERMINED BY QUANTITATIVE REAL-TIME PCR (QPCR) AND WESTERN BLOT, RESPECTIVELY. IMMUNOHISTOCHEMISTRY STAINING IN THE KIDNEY TISSUES FROM MICE OR PATIENTS WAS USED TO EVALUATE PROTEIN LEVELS. FLOW CYTOMETRY WAS USED TO ANALYZE THE CELL CYCLE DISTRIBUTIONS AND APOPTOSIS. RNA-SEQUENCING WAS PERFORMED FOR DIFFERENTIAL EXPRESSION GENES IN THE KIDNEY TISSUES OF THE MEN1F/F AND MEN1?/? MICE. CHROMATIN IMMUNOPRECIPITATION SEQUENCING (CHIP-SEQ) WAS CARRIED OUT FOR IDENTIFICATION OF MENIN- AND H3K4ME3-ENRICHED REGIONS WITHIN THE WHOLE GENOME IN THE MOUSE KIDNEY TISSUE. CHIP-QPCR ASSAYS WERE PERFORMED FOR OCCUPANCY OF MENIN AND H3K4ME3 AT THE GENE PROMOTER REGIONS. LUCIFERASE REPORTER ASSAY WAS USED TO DETECT THE PROMOTER ACTIVITY. THE EXACERBATED UNILATERAL URETERAL OBSTRUCTION (UUO) MODELS IN THE MEN1F/F AND MEN1?/? MICE WERE USED TO ASSESS THE PHARMACOLOGICAL EFFECTS OF RH-HGF ON RENAL FIBROSIS. RESULTS: THE EXPRESSION OF MEN1 IS REDUCE IN KIDNEY TISSUES OF FIBROTIC MOUSE AND HUMAN DIABETIC PATIENTS AND TREATMENT WITH FIBROTIC FACTOR RESULTS IN THE DOWNREGULATION OF MEN1 EXPRESSION IN RENAL TUBULAR EPITHELIAL CELLS (RTECS). DISRUPTION OF MEN1 IN RTECS LEADS TO HIGH EXPRESSION OF ALPHA-SMA AND COLLAGEN 1, WHEREAS MEN1 OVEREXPRESSION RESTRAINS EPITHELIAL-TO-MESENCHYMAL TRANSITION (EMT) INDUCED BY TGF-BETA TREATMENT. CONDITIONAL KNOCKOUT OF MEN1 RESULTED IN CHRONIC RENAL FIBROSIS AND UUO-INDUCED TUBULOINTERSTITIAL FIBROSIS (TIF), WHICH IS ASSOCIATED WITH AN INCREASED INDUCTION OF EMT, G2/M ARREST AND JNK SIGNALING. MECHANISTICALLY, MENIN RECRUITS AND INCREASES H3K4ME3 AT THE PROMOTER REGIONS OF HEPATOCYTE GROWTH FACTOR (HGF) AND A DISINTEGRIN AND METALLOPROTEINASE WITH THROMBOSPONDIN MOTIFS 5 (ADAMTS5) GENES AND ENHANCES THEIR TRANSCRIPTIONAL ACTIVATION. IN THE UUO MICE MODEL, EXOGENOUS HGF RESTORED THE EXPRESSION OF ADAMTS5 AND AMELIORATED RENAL FIBROSIS INDUCED BY MEN1 DEFICIENCY. CONCLUSIONS: THESE FINDINGS DEMONSTRATE THAT MEN1 IS AN ESSENTIAL ANTIFIBROTIC FACTOR IN RENAL FIBROGENESIS AND COULD BE A POTENTIAL TARGET FOR ANTIFIBROTIC THERAPY. 2022 18 1792 29 EFFECT OF CHRONIC WESTERN DIETS ON NON-ALCOHOLIC FATTY LIVER OF MALE MICE MODIFYING THE PPAR-GAMMA PATHWAY VIA MIR-27B-5P REGULATION. WESTERN DIETS CONTRIBUTE TO METABOLIC DISEASES. HOWEVER, THE EFFECTS OF VARIOUS DIETS AND EPIGENETIC MECHANISMS ARE MOSTLY UNKNOWN. HERE, SIX WEEK-OLD C57BL/6J MALE AND FEMALE MICE WERE FED WITH A LOW-FAT DIET (LFD), HIGH-FAT DIET (HFD), AND HIGH-FAT HIGH-FRUCTOSE DIET (HFD-HF) FOR 20 WEEKS. WE DETERMINED THAT HFD-HF OR HFD MICE EXPERIENCED SIGNIFICANT METABOLIC DYSREGULATION COMPARED TO THE LFD. HFD-HF AND HFD-FED MALE MICE SHOWED SIGNIFICANTLY INCREASED BODY WEIGHT, LIVER SIZE, AND FASTING GLUCOSE LEVELS WITH DOWNREGULATED PPARGAMMA, SCD1, AND FAS PROTEIN EXPRESSION. IN CONTRAST, FEMALE MICE WERE LESS AFFECTED BY HFD AND HFD-HF. AS MIR-27B CONTAINS A SEED SEQUENCE IN PPARGAMMA, IT WAS DISCOVERED THAT THESE CHANGES ARE ACCOMPANIED BY MALE-SPECIFIC UPREGULATION OF MIR-27B-5P, WHICH IS EVEN MORE PRONOUNCED IN THE HFD-HF GROUP (P < 0.01 VS. LFD) COMPARED TO THE HFD GROUP (P < 0.05 VS. LFD). OTHER MIR-27 SUBTYPES WERE INCREASED BUT NOT SIGNIFICANTLY. HFD-HF SHOWED INSIGNIFICANT CHANGES IN FIBROSIS MARKERS WHEN COMPARED TO LFD. INTERESTINGLY, FAT BALLOONING IN HEPATOCYTES WAS INCREASED IN HFD-FED MICE COMPARED TO HFD-HF FED MICE, HOWEVER, THE HFD-HF LIVER SHOWED AN INCREASE IN THE NUMBER OF SMALL CELLS. HERE, WE CONCLUDED THAT CHRONIC WESTERN DIET-COMPOSITION ADMINISTERED FOR 20 WEEKS MAY SURPASS THE NON-ALCOHOLIC FATTY LIVER (NAFL) STAGE BUT MAY BE AT AN INTERMEDIATE STAGE BETWEEN FATTY LIVER AND FIBROSIS VIA MIR-27B-5P-INDUCED PPARGAMMA DOWNREGULATION. 2021 19 3306 31 HIGH-PHOSPHATE-INDUCED CALCIFICATION IS RELATED TO SM22ALPHA PROMOTER METHYLATION IN VASCULAR SMOOTH MUSCLE CELLS. HYPERPHOSPHATEMIA IS CLOSELY RELATED TO VASCULAR CALCIFICATION IN PATIENTS WITH CHRONIC KIDNEY DISEASE. VASCULAR SMOOTH MUSCLE CELLS (VSMCS) EXPOSED TO HIGH PHOSPHATE CONCENTRATIONS IN VITRO UNDERGO PHENOTYPIC TRANSITION TO OSTEOBLAST-LIKE CELLS. MECHANISMS UNDERLYING THIS TRANSDIFFERENTIATION ARE NOT CLEAR. IN THIS STUDY WE USED TWO IN VITRO MODELS, HUMAN AORTIC SMOOTH MUSCLE CELLS AND RAT AORTIC RINGS, TO INVESTIGATE THE PHENOTYPIC TRANSITION OF VSMCS INDUCED BY HIGH PHOSPHATE. WE FOUND THAT HIGH PHOSPHATE CONCENTRATION (3.3 MMOL/L) IN THE MEDIUM WAS ASSOCIATED WITH INCREASED DNA METHYLTRANSFERASE ACTIVITY AND METHYLATION OF THE PROMOTER REGION OF SM22ALPHA. THIS WAS ACCOMPANIED BY LOSS OF THE SMOOTH MUSCLE CELL-SPECIFIC PROTEIN SM22ALPHA, GAIN OF THE OSTEOBLAST TRANSCRIPTION FACTOR CBFA1, AND INCREASED ALKALINE PHOSPHATASE ACTIVITY WITH THE SUBSEQUENT IN VITRO CALCIFICATION. THE ADDITION OF A DEMETHYLATING AGENT (PROCAINE) TO THE HIGH-PHOSPHATE MEDIUM REDUCED DNA METHYLTRANSFERASE ACTIVITY AND PREVENTED METHYLATION OF THE SM22ALPHA PROMOTER, WHICH WAS ACCOMPANIED BY AN INCREASE IN SM22ALPHA EXPRESSION AND LESS CALCIFICATION. ADDITIONALLY, DOWNREGULATION OF SM22ALPHA, EITHER BY SIRNA OR BY A METHYL GROUP DONOR (S-ADENOSYL METHIONINE), RESULTED IN OVEREXPRESSION OF CBFA1. IN CONCLUSION, WE DEMONSTRATE THAT METHYLATION OF SM22ALPHA PROMOTER IS AN IMPORTANT EVENT IN VASCULAR SMOOTH MUSCLE CELL CALCIFICATION AND THAT HIGH PHOSPHATE INDUCES THIS EPIGENETIC MODIFICATION. THESE FINDINGS UNCOVER A NEW INSIGHT INTO MECHANISMS BY WHICH HIGH PHOSPHATE CONCENTRATION PROMOTES VASCULAR CALCIFICATION. 2010 20 3302 28 HIGH-FAT, SUCROSE AND SALT-RICH DIET DURING RAT SPERMATOGENESIS LEAD TO THE DEVELOPMENT OF CHRONIC KIDNEY DISEASE IN THE FEMALE OFFSPRING OF THE F2 GENERATION. EFFECTS OF FEEDING MALE RATS DURING SPERMATOGENESIS A HIGH-FAT, HIGH-SUCROSE AND HIGH-SALT DIET (HFSSD) OVER TWO GENERATIONS (F0 AND F1) ON RENAL OUTCOMES ARE UNKNOWN. MALE F0 AND F1 RATS WERE FED EITHER CONTROL DIET (F0CD+F1CD) OR HFSSD (F0HD+F1HD). THE OUTCOMES WERE GLOMERULAR FILTRATION RATE AND URINARY ALBUMIN EXCRETION IN F1 AND F2 OFFSPRING. IF BOTH OUTCOMES WERE ALTERED A MORPHOLOGICAL AND MOLECULAR ASSESSMENT WAS DONE. F2 OFFSPRING OF BOTH SEXES HAD A DECREASED GFR. HOWEVER, INCREASED URINARY ALBUMIN EXCRETION WAS ONLY OBSERVED IN FEMALE F2 F0HD+F1HD OFFSPRING COMPARED WITH CONTROLS. F0HD+F1HD FEMALE F2 OFFSPRING DEVELOPED GLOMERULOSCLEROSIS (+31%; P < .01) AND INCREASED RENAL INTERSTITIAL FIBROSIS (+52%; P < .05). RNA SEQUENCING FOLLOWED BY QRT-PCR VALIDATION SHOWED THAT FOUR GENES (ENPP6, TMEM144, CD300LF, AND ACTR3B) WERE DIFFERENTIALLY REGULATED IN THE KIDNEYS OF FEMALE F2 OFFSPRING. LNCRNA XR-146683.1 EXPRESSION DECREASED IN FEMALE F0HD+F1HD F2 OFFSPRING AND ITS EXPRESSION WAS (R = 0.44, P = .027) CORRELATED WITH THE EXPRESSION OF TMEM144. METHYLATION OF CPG ISLANDS IN THE PROMOTER REGION OF THE CD300LF GENE WAS INCREASED (P = .001) IN FEMALE F2 F0HD+F1HD OFFSPRING COMPARED TO CONTROLS. PROMOTER CPG ISLAND METHYLATION RATE OF CD300LF WAS INVERSELY CORRELATED WITH CD300LF MRNA EXPRESSION IN F2 FEMALE OFFSPRING (R = -0.483, P = .012). CD300LF MRNA EXPRESSION WAS INVERSELY CORRELATED WITH THE URINARY ALBUMIN-TO-CREATININE RATIO IN FEMALE F2 OFFSPRING (R = -0.588, P = .005). PATERNAL PRE-CONCEPTIONAL UNHEALTHY DIET GIVEN FOR TWO GENERATIONS PREDISPOSE FEMALE F2 OFFSPRING TO CHRONIC KIDNEY DISEASE DUE TO EPIGENETIC ALTERATIONS OF RENAL GENE EXPRESSION. PARTICULARLY, CD300LF GENE PROMOTOR METHYLATION WAS INVERSELY ASSOCIATED WITH CD300LF MRNA EXPRESSION AND CD300LF MRNA EXPRESSION ITSELF WAS INVERSELY ASSOCIATED WITH URINARY ALBUMIN EXCRETION IN F2 FEMALE OFFSPRING WHOSE FATHERS AND GRANDFATHERS GOT A PRE-CONCEPTIONAL UNHEALTHY DIET. 2022