1 4731 173 NOVEL ANTI-AGING GENE NM_026333 CONTRIBUTES TO PROTON-INDUCED AGING VIA NCX1-PATHWAY. DIET-INDUCED METABOLIC ACIDOSIS IS ASSOCIATED WITH THE IMPAIRMENT OF BONE METABOLISM AND AN INCREASED RISK OF A NUMBER OF CHRONIC NONCOMMUNICABLE DISEASES, SUCH AS TYPE 2 DIABETES MELLITUS AND HYPERTENSION. LOW SERUM BICARBONATE IS ASSOCIATED WITH HIGH MORTALITY IN HEALTHY OLDER INDIVIDUALS. RECENTLY, WE DEMONSTRATED THAT BOTH COUPLING FACTOR 6 (CF6)-OVEREXPRESSING TRANSGENIC (TG) AND HIGH SALT-FED MICE WHICH HAD SUSTAINED INTRACELLULAR ACIDOSIS, DUE TO ENHANCED PROTON IMPORT THROUGH ECTO-F(1)F(O) COMPLEX AND/OR REDUCED PROTON EXPORT THROUGH NA(+)-K(+) ATPASE INHIBITION, DISPLAYED SHORTENED LIFESPAN AND EARLY SENESCENCE-ASSOCIATED PHENOTYPES SUCH AS SIGNS OF HAIR GREYING AND ALOPECIA, WEIGHT LOSS, AND/OR REDUCED ORGAN MASS. IN THIS STUDY, WE SEARCHED CAUSATIVE GENES OF PROTON-INDUCED AGING IN CF6-OVEREXPRESSING TG AND HIGH SALT-FED MICE. WE DISCOVERED NM_026333 AS A NOVEL ANTI-AGING GENE WHICH WAS DOWNREGULATED IN THE HEART AND KIDNEY IN BOTH TYPES OF MICE. NM_026333 PROTEIN CONSISTS OF 269 AMINO ACIDS WITH TRANSMEMBRANE REGION (90-193AA). INDUCTION OF NM_026333 OR RECOMBINANT PROTEIN RESCUED TG CELLS AND CF6-TREATED HUMAN CELLS FROM AGING HALLMARKS OF IMPAIRED AUTOPHAGY, GENOMIC INSTABILITY, AND EPIGENETIC ALTERATION. NM_026333 PROTEIN DIRECTLY BOUND PLASMA MEMBRANE NA(+)-CA(2+) EXCHANGER 1 (NCX1) TO SUPPRESS ITS REVERSE MODE, AND CANCELLED PROTON-INDUCED EPIGENETIC REGRESSION OF ATG7 THAT WAS CAUSED BY H3K4 AND H4K20 TRI-METHYLATION VIA SUPPRESSION OF DEMETHYLASE AND H4K5 ACETYLATION VIA SUPPRESSION OF NUCLEAR HDAC3-HDAC4-EMERIN SYSTEM. NM_026333 ALSO ATTENUATED PROTON-INDUCED IMPAIRED FORMATION OF AUTOLYSOSOME, AN INCREASE IN NUCLEAR ACETYLATED LC3 II, AND ACETYLATION OF ATG7. THESE EFFECTS REAPPEARED BY NCX1 INHIBITOR. FURTHERMORE, NCX1 INHIBITOR EXTENDED LIFESPAN COMPARED WITH VEHICLE-TREATMENT IN TG MICE. THIS STUDY WILL SHED LIGHT ON NOVEL AGING MECHANISM AND PROVIDE IMPLICATIONS IN A TARGET FOR ANTI-AGING THERAPY. 2018 2 3807 103 INTRACELLULAR PROTONS ACCELERATE AGING AND SWITCH ON AGING HALLMARKS IN MICE. DIET-INDUCED METABOLIC ACIDOSIS IS ASSOCIATED WITH THE IMPAIRMENT OF BONE METABOLISM AND AN INCREASED RISK OF A NUMBER OF CHRONIC NONCOMMUNICABLE DISEASES, SUCH AS TYPE 2 DIABETES MELLITUS AND HYPERTENSION. THE SERUM BICARBONATE LEVEL IS AN INDEPENDENT PREDICTOR OF CHRONIC KIDNEY DISEASE PROGRESSION. WE INVESTIGATED WHETHER PROTON ACCELERATES AGING BY ANALYZING BOTH COUPLING FACTOR 6-OVEREXPRESSING TRANSGENIC (TG) AND HIGH SALT-FED MICE WHICH DISPLAY SUSTAINED INTRACELLULAR ACIDOSIS, DUE TO ENHANCED PROTON IMPORT THROUGH ECTO-F(1) F(O) COMPLEX AND/OR REDUCED PROTON EXPORT THROUGH NA(+) -K(+) ATPASE INHIBITION. BOTH TYPES OF MICE DISPLAYED SHORTENED LIFESPAN AND EARLY SENESCENCE-ASSOCIATED PHENOTYPES SUCH AS SIGNS OF HAIR GREYING AND ALOPECIA, WEIGHT LOSS, AND/OR REDUCED ORGAN MASS. IN CHRONIC INTRACELLULAR ACIDOSIS MICE, AUTOPHAGY WAS IMPAIRED BY REGRESSION OF ATG7, AN INCREASE IN NUCLEAR ACETYLATED LC3 II, AND ACETYLATION OF ATG7. THE INCREASE IN HISTONE 3 TRIMETHYLATION AT LYSINE 4 (H3K4ME3) AND H4K20ME3 AND THE DECREASE IN H3K9ME3 AND H3K27ME3 WERE OBSERVED IN THE HEART AND KIDNEY OBTAINED FROM BOTH TG AND HIGH SALT-FED MICE. THE DECREASE IN LAMIN A/C, EMERIN, AND HETEROCHROMATIN PROTEIN 1ALPHA WITHOUT CHANGES IN BARRIER-TO-AUTOINTEGRATION FACTOR AND HIGH-MOBILITY GROUP BOX 1 WAS CONFIRMED IN TG AND HIGH SALT-FED MICE. SUPPRESSION OF NUCLEAR HISTONE DEACETYLASE 3-EMERIN SYSTEM IS ATTRIBUTABLE TO EPIGENETIC REGRESSION OF ATG7 AND H4K5 ACETYLATION. THESE FINDINGS WILL SHED LIGHT ON NOVEL AGING AND IMPAIRED AUTOPHAGY MECHANISM, AND PROVIDE IMPLICATIONS IN A TARGET FOR ANTIAGING THERAPY. 2018 3 1104 29 COMBINED EXPOSURE TO PROTONS AND (56)FE LEADS TO OVEREXPRESSION OF IL13 AND REACTIVATION OF REPETITIVE ELEMENTS IN THE MOUSE LUNG. INTEREST IN DEEP SPACE EXPLORATION UNDERLINES THE NEEDS TO INVESTIGATE THE EFFECTS OF EXPOSURE TO COMBINED SOURCES OF SPACE RADIATION. THE LUNG IS A TARGET ORGAN FOR RADIATION, AND EXPOSURE TO PROTONS AND HEAVY IONS AS RADIATION SOURCES MAY LEAD TO THE DEVELOPMENT OF DEGENERATIVE DISEASE AND CANCER. IN THIS STUDY, WE EVALUATED THE PRO-FIBROTIC AND EPIGENETIC EFFECTS OF EXPOSURE TO PROTONS (150 MEV/NUCLEON, 0.1 GY) AND HEAVY IRON IONS ((56)FE, 600 MEV/NUCLEON, 0.5 GY) ALONE OR IN COMBINATION (PROTONS ON DAY 1 AND (56)FE ON DAY 2) IN C57BL/6 MALE MICE 4 WEEKS AFTER IRRADIATION. EXPOSURE TO (56)FE, PROTON OR IN COMBINATION, DID NOT RESULT IN HISTOPATHOLOGICAL CHANGES IN THE MURINE LUNG. AT THE SAME TIME, COMBINED EXPOSURE TO PROTONS AND (56)FE RESULTED IN PRONOUNCED MOLECULAR ALTERATIONS IN COMPARISON WITH EITHER SOURCE OF RADIATION ALONE. SPECIFICALLY, WE OBSERVED A SUBSTANTIAL INCREASE IN THE EXPRESSION OF CYTOKINE IL13, LOSS OF EXPRESSION OF DNA METHYLTRANSFERASE DNMT1, AND REACTIVATION OF LINE-1, SINE B1 RETROTRANSPOSONS, AND MAJOR AND MINOR SATELLITES. GIVEN THE DELETERIOUS POTENTIAL OF THE OBSERVED EFFECTS THAT MAY LEAD TO DEVELOPMENT OF CHRONIC LUNG INJURY, PULMONARY FIBROSIS, AND CANCER, FUTURE STUDIES DEVOTED TO THE INVESTIGATION OF THE LONG-TERM EFFECTS OF COMBINED EXPOSURES TO PROTON AND HEAVY IONS ARE CLEARLY NEEDED. 2015 4 3237 41 HEPATIC COX-2 EXPRESSION PROTECTS MICE FROM AN ALCOHOL-HIGH FAT DIET-INDUCED METABOLIC DISORDER BY INVOLVING PROTEIN ACETYLATION RELATED ENERGY METABOLISM. PURPOSE: A DIET HIGH IN FAT AND ETHANOL OFTEN RESULTS IN CHRONIC METABOLIC DISORDER, HEPATIC STEATOSIS, AND LIVER INFLAMMATION. CONSTITUTIVE HEPATIC CYCLOOXYGENASE-2 (COX-2) EXPRESSION COULD PROTECT FROM HIGH FAT-INDUCED METABOLISM DISTURBANCE IN A MURINE MODEL. IN THIS STUDY, WE EXPLORED THE INFLUENCE OF HCOX-2 TRANSGENIC [TG] TO HIGH FAT WITH ETHANOL-INDUCED METABOLIC DISORDER AND LIVER INJURY USING A MOUSE ANIMAL MODEL. METHODS: 12-WEEK-OLD MALE HEPATIC HCOX-2 TRANSGENIC (TG) OR WILD TYPE MICE (WT) WERE FED EITHER A HIGH FAT AND ETHANOL LIQUID DIET (HF+ETH) OR A REGULAR CONTROL DIET (RCD) FOR 5 WEEKS (FOUR GROUPS: RCD/WT, RCD/TG; HF+ETH/TG, HF+ETH/WT). WE ASSESSED METABOLIC BIOMARKERS, CYTOKINE PROFILES, HISTOMORPHOLOGY, AND GENE EXPRESSION TO STUDY THE IMPACT OF PERSISTENT HEPATIC COX-2 EXPRESSION ON DIET-INDUCED LIVER INJURY. RESULTS: IN THE HF+ETH DIET, CONSTITUTIVELY HEPATIC HUMAN COX-2 EXPRESSION PROTECTS MICE FROM BODY WEIGHT GAIN AND WHITE ADIPOSE TISSUE ACCUMULATION, ACCOMPANIED BY IMPROVED IPGTT RESPONSE, SERUM TRIGLYCERIDE/CHOLESTEROL LEVELS, AND LOWER LEVELS OF SERUM AND LIVER INFLAMMATORY CYTOKINES. HISTOLOGICALLY, HCOX-2 MICE SHOWED DECREASED HEPATIC LIPID DROPLETS ACCUMULATION, DECREASED HEPATOCYTE BALLOONING, AND IMPROVED STEATOSIS SCORES. HEPATIC HCOX-2 OVEREXPRESSION ENHANCED AKT INSULIN SIGNALING AND INCREASED FATTY ACID SYNTHESIS IN BOTH RCD AND HF+ETH DIET GROUPS. THE ANTI-LIPOGENIC EFFECT OF HCOX-2 TG IN THE HF+ETH DIET ANIMALS WAS MEDIATED BY INCREASING LIPID DISPOSAL THROUGH ENHANCED BETA-OXIDATION VIA ELEVATIONS IN THE EXPRESSION OF PPARALPHA AND PPARGAMMA, AND INCREASED HEPATIC AUTOPHAGY AS ASSESSED BY THE RATIO OF AUTOPHAGY MARKERS LC3 II/I IN HEPATIC TISSUE. VARIOUS PROTEIN ACETYLATION PATHWAY COMPONENTS, INCLUDING HAT, HDAC1, SIRT1, AND SNAIL1, WERE MODULATED IN HCOX-2 TG MICE IN EITHER RCD OR HF+ETH DIET. CONCLUSIONS: HEPATIC HUMAN COX-2 EXPRESSION PROTECTED MICE FROM THE METABOLIC DISORDER AND LIVER INJURY INDUCED BY A HIGH FAT AND ETHANOL DIET BY ENHANCING HEPATIC LIPID EXPENDITURE. EPIGENETIC REPROGRAMMING OF DIVERSE METABOLIC GENES MIGHT BE INVOLVED IN THE ANTI-LIPOGENIC EFFECT OF COX-2. 2021 5 3096 36 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 6 3390 32 HOPX PLAYS A CRITICAL ROLE IN ANTIRETROVIRAL DRUGS INDUCED EPIGENETIC MODIFICATION AND CARDIAC HYPERTROPHY. PEOPLE LIVING WITH HIV (PLWH) HAVE TO TAKE AN ANTIRETROVIRAL THERAPY (ART) FOR LIFE AND SHOW NONCOMMUNICABLE ILLNESSES SUCH AS CHRONIC INFLAMMATION, IMMUNE ACTIVATION, AND MULTIORGAN DYSREGULATION. RECENT STUDIES SUGGEST THAT LONG-TERM USE OF ART INDUCES COMORBID CONDITIONS AND IS ONE OF THE LEADING CAUSES OF HEART FAILURE IN PLWH. HOWEVER, THE MOLECULAR MECHANISM OF ANTIRETROVIRAL DRUGS (ARVS) INDUCED HEART FAILURE IS UNCLEAR. TO DETERMINE THE MECHANISM OF ARVS INDUCED CARDIAC DYSFUNCTION, WE PERFORMED GLOBAL TRANSCRIPTOMIC PROFILING OF ARVS TREATED NEONATAL RAT VENTRICULAR CARDIOMYOCYTES IN CULTURE. DIFFERENTIALLY EXPRESSED GENES WERE IDENTIFIED BY RNA-SEQUENCING. OUR DATA SHOW THAT ARVS TREATMENT CAUSES UPREGULATION OF SEVERAL BIOLOGICAL FUNCTIONS ASSOCIATED WITH CARDIOTOXICITY, HYPERTROPHY, AND HEART FAILURE. GLOBAL GENE EXPRESSION DATA WERE VALIDATED IN CARDIAC TISSUE ISOLATED FROM HIV PATIENTS HAVING A HISTORY OF ART. INTERESTINGLY, WE FOUND THAT HOMEODOMAIN-ONLY PROTEIN HOMEOBOX (HOPX) EXPRESSION WAS SIGNIFICANTLY INCREASED IN CARDIOMYOCYTES TREATED WITH ARVS AND IN THE HEART TISSUE OF HIV PATIENTS. FURTHERMORE, WE FOUND THAT HOPX PLAYS A CRUCIAL ROLE IN ARVS MEDIATED CELLULAR HYPERTROPHY. MECHANISTICALLY, WE FOUND THAT HOPX PLAYS A CRITICAL ROLE IN EPIGENETIC REGULATION, THROUGH DEACETYLATION OF HISTONE, WHILE THE HDAC INHIBITOR, TRICHOSTATIN A, CAN RESTORE THE ACETYLATION LEVEL OF HISTONE 3 IN THE PRESENCE OF ARVS. 2021 7 5474 33 RESTORATION OF HISTONE ACETYLATION AMELIORATES DISEASE AND METABOLIC ABNORMALITIES IN A FUS MOUSE MODEL. DYSREGULATION OF EPIGENETIC MECHANISMS IS EMERGING AS A CENTRAL EVENT IN NEURODEGENERATIVE DISORDERS, INCLUDING AMYOTROPHIC LATERAL SCLEROSIS (ALS). IN MANY MODELS OF NEURODEGENERATION, GLOBAL HISTONE ACETYLATION IS DECREASED IN THE AFFECTED NEURONAL TISSUES. HISTONE ACETYLATION IS CONTROLLED BY THE ANTAGONISTIC ACTIONS OF TWO PROTEIN FAMILIES -THE HISTONE ACETYLTRANSFERASES (HATS) AND THE HISTONE DEACETYLASES (HDACS). DRUGS INHIBITING HDAC ACTIVITY ARE ALREADY USED IN THE CLINIC AS ANTI-CANCER AGENTS. THE AIM OF THIS STUDY WAS TO EXPLORE THE THERAPEUTIC POTENTIAL OF HDAC INHIBITION IN THE CONTEXT OF ALS. WE DISCOVERED THAT TRANSGENIC MICE OVEREXPRESSING WILD-TYPE FUS ("TG FUS+/+"), WHICH RECAPITULATE MANY ASPECTS OF HUMAN ALS, SHOWED REDUCED GLOBAL HISTONE ACETYLATION AND ALTERATIONS IN METABOLIC GENE EXPRESSION, RESULTING IN A DYSREGULATED METABOLIC HOMEOSTASIS. CHRONIC TREATMENT OF TG FUS+/+ MICE WITH ACY-738, A POTENT HDAC INHIBITOR THAT CAN CROSS THE BLOOD-BRAIN BARRIER, AMELIORATED THE MOTOR PHENOTYPE AND SUBSTANTIALLY EXTENDED THE LIFE SPAN OF THE TG FUS+/+ MICE. AT THE MOLECULAR LEVEL, ACY-738 RESTORED GLOBAL HISTONE ACETYLATION AND METABOLIC GENE EXPRESSION, THEREBY RE-ESTABLISHING METABOLITE LEVELS IN THE SPINAL CORD. TAKEN TOGETHER, OUR FINDINGS LINK EPIGENETIC ALTERATIONS TO METABOLIC DYSREGULATION IN ALS PATHOLOGY, AND HIGHLIGHT ACY-738 AS A POTENTIAL THERAPEUTIC STRATEGY TO TREAT THIS DEVASTATING DISEASE. 2019 8 6000 33 THE ACTIVATION OF HISTONE DEACETYLASES 4 PREVENTED ENDOTHELIAL DYSFUNCTION: A CRUCIAL MECHANISM OF HUANGQIGUIZHIWUWU DECOCTION IN IMPROVING MICROCIRCULATION DYSFUNCTION IN DIABETES. ETHNOPHARMACOLOGICAL RELEVANCE: THE REGULATION OF EPIGENETIC FACTORS IS CONSIDERED A CRUCIAL TARGET FOR SOLVING COMPLEX CHRONIC DISEASES SUCH AS CARDIO-CEREBROVASCULAR DISEASES. HUANGQIGUIZHIWUWU DECOCTION (HGWWD), A CLASSIC CHINESE PRESCRIPTION, IS MAINLY USED TO TREAT VARIOUS VASCULAR DISEASES. ALTHOUGH OUR PREVIOUS STUDIES REPORTED THAT HGWWD COULD EFFECTIVELY PREVENT VASCULAR DYSFUNCTION IN DIABETIC RODENT MODELS, THE PRECISE MECHANISM IS STILL ELUSIVE. AIM OF THE STUDY: IN THIS STUDY, WE INVESTIGATED THE EPIGENETIC MECHANISMS OF MODULATING THE DAMAGE OF VASCULAR ENDOTHELIAL CELLS IN DIABETES BY HGWWD. METHODS: WE FIRST ANALYZED COMMON ACTIVE COMPONENTS OF HGWWD BY USING HPLC-Q-TOF-MS/MS ANALYSIS, AND PREDICTED THE ISOFORMS OF HISTONE DEACETYLASE (HDAC) THAT CAN POTENTIALLY COMBINE THE ABOVE ACTIVE COMPONENTS BY SYSTEMS PHARMACOLOGY. NEXT, WE SCREENED THE INVOLVEMENT OF SPECIFIC HDAC ISOFORMS IN THE PROTECTIVE EFFECT OF HGWWD ON VASCULAR INJURY BY USING PHARMACOLOGICAL BLOCKADE COMBINED WITH THE EVALUATION OF VASCULAR FUNCTION IN VIVO AND IN VITRO. RESULTS: FIRSTLY, HDAC1, HDAC2, HDAC3, HDAC4, HDAC6, HDAC7, SIRT2, AND SIRT3 HAVE BEEN IMPLICATED WITH THE POSSIBILITY OF BINDING TO THE THIRTY-ONE COMMON ACTIVE COMPONENTS IN HGWWD. FURTHERMORE, THE PROTECTIVE EFFECT OF HGWWD IS REVERSED BY BOTH TSA (HDAC INHIBITOR) AND MC1568 (CLASS II HDAC INHIBITOR) ON VASCULAR IMPAIRMENT ACCOMPANIED BY REDUCED AORTIC HDAC ACTIVITY IN STZ MICE. FINALLY, INHIBITION OF HDAC4 BLOCKED THE PROTECTIVE EFFECT OF HGWWD ON MICROVASCULAR AND ENDOTHELIAL DYSFUNCTION IN DIABETIC MICE. CONCLUSIONS: THESE RESULTS PROVE THE KEY ROLE OF HDAC4 IN DIABETES-INDUCED MICROVASCULAR DYSFUNCTION AND UNDERLYING EPIGENETIC MECHANISMS FOR THE PROTECTIVE EFFECT OF HGWWD IN DIABETES. 2023 9 26 33 A 6-ALKYLSALICYLATE HISTONE ACETYLTRANSFERASE INHIBITOR INHIBITS HISTONE ACETYLATION AND PRO-INFLAMMATORY GENE EXPRESSION IN MURINE PRECISION-CUT LUNG SLICES. LYSINE ACETYLATIONS ARE POST-TRANSLATIONAL MODIFICATIONS OF CELLULAR PROTEINS, THAT ARE CRUCIAL IN THE REGULATION OF MANY CELLULAR PROCESSES. LYSINE ACETYLATIONS ON HISTONE PROTEINS ARE PART OF THE EPIGENETIC CODE REGULATING GENE EXPRESSION AND ARE INSTALLED BY HISTONE ACETYLTRANSFERASES. OBSERVATIONS THAT INFLAMMATORY LUNG DISEASES, SUCH AS ASTHMA AND CHRONIC OBSTRUCTIVE PULMONARY DISEASE, ARE CHARACTERIZED BY INCREASED HISTONE ACETYLTRANSFERASE ACTIVITY INDICATE THAT DEVELOPMENT OF SMALL MOLECULE INHIBITORS FOR THESE ENZYMES MIGHT BE A VALUABLE APPROACH TOWARDS NEW THERAPIES FOR THESE DISEASES. THE 6-ALKYLSALICYLATE MG149 IS A CANDIDATE TO EXPLORE THIS HYPOTHESIS BECAUSE IT HAS BEEN DEMONSTRATED TO INHIBIT THE MYST TYPE HISTONE ACETYLTRANSFERASES. IN THIS STUDY, WE DETERMINED THE K(I) VALUE FOR INHIBITION OF THE MYST TYPE HISTONE ACETYLTRANSFERASE KAT8 BY MG149 TO BE 39 +/- 7.7 MUM. UPON INVESTIGATING WHETHER THE INHIBITION OF HISTONE ACETYLTRANSFERASES BY MG149 CORRELATES WITH INHIBITION OF HISTONE ACETYLATION IN MURINE PRECISION-CUT LUNG SLICES, INHIBITION OF ACETYLATION WAS OBSERVED USING AN LC-MS/MS BASED ASSAY ON HISTONE H4 RES 4-17, WHICH CONTAINS THE TARGET LYSINE OF KAT8. FOLLOWING UP ON THIS, UPON TREATMENT WITH MG149, REDUCED PRO-INFLAMMATORY GENE EXPRESSION WAS OBSERVED IN LIPOPOLYSACCHARIDE AND INTERFERON GAMMA STIMULATED MURINE PRECISION-CUT LUNG SLICES. BASED ON THIS, WE PROPOSE THAT 6-ALKYLSALICYLATES SUCH AS MG149 HAVE POTENTIAL FOR DEVELOPMENT TOWARDS APPLICATIONS IN THE TREATMENT OF INFLAMMATORY LUNG DISEASES. 2017 10 3327 32 HISTONE DEACETYLASE 4 PROMOTES CHOLESTATIC LIVER INJURY IN THE ABSENCE OF PROHIBITIN-1. PROHIBITIN-1 (PHB1) IS AN EVOLUTIONARILY CONSERVED PLEIOTROPIC PROTEIN THAT PARTICIPATES IN DIVERSE PROCESSES DEPENDING ON ITS SUBCELLULAR LOCALIZATION AND INTERACTOME. RECENT DATA HAVE INDICATED A DIVERSE ROLE FOR PHB1 IN THE PATHOGENESIS OF OBESITY, CANCER, AND INFLAMMATORY BOWEL DISEASE, AMONG OTHERS. DATA PRESENTED HERE SUGGEST THAT PHB1 IS ALSO LINKED TO CHOLESTATIC LIVER DISEASE. EXPRESSION OF PHB1 IS MARKEDLY REDUCED IN PATIENTS WITH PRIMARY BILIARY CIRRHOSIS AND BILIARY ATRESIA OR WITH ALAGILLE SYNDROME, TWO MAJOR PEDIATRIC CHOLESTATIC CONDITIONS. IN THE EXPERIMENTAL MODEL OF BILE DUCT LIGATION, SILENCING OF PHB1 INDUCED LIVER FIBROSIS, REDUCED ANIMAL SURVIVAL, AND INDUCED BILE DUCT PROLIFERATION. IMPORTANTLY, THE MODULATORY EFFECT OF PHB1 IS NOT DEPENDENT ON ITS KNOWN MITOCHONDRIAL FUNCTION. ALSO, PHB1 INTERACTS WITH HISTONE DEACETYLASE 4 (HDAC4) IN THE PRESENCE OF BILE ACIDS. HENCE, PHB1 DEPLETION LEADS TO INCREASED NUCLEAR HDAC4 CONTENT AND ITS ASSOCIATED EPIGENETIC CHANGES. REMARKABLY, HDAC4 SILENCING AND THE ADMINISTRATION OF THE HDAC INHIBITOR PARTHENOLIDE DURING OBSTRUCTIVE CHOLESTASIS IN VIVO PROMOTE GENOMIC REPROGRAMMING, LEADING TO REGRESSION OF THE FIBROTIC PHENOTYPE IN LIVER-SPECIFIC PHB1 KNOCKOUT MICE. CONCLUSION: PHB1 IS AN IMPORTANT MEDIATOR OF CHOLESTATIC LIVER INJURY THAT REGULATES THE ACTIVITY OF HDAC4, WHICH CONTROLS SPECIFIC EPIGENETIC MARKERS; THESE RESULTS IDENTIFY POTENTIAL NOVEL STRATEGIES TO TREAT LIVER INJURY AND FIBROSIS, PARTICULARLY AS A CONSEQUENCE OF CHRONIC CHOLESTASIS. 2015 11 5636 38 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 12 1335 34 DERMAL FIBROBLASTS CULTURED FROM DONORS WITH TYPE 2 DIABETES MELLITUS RETAIN AN EPIGENETIC MEMORY ASSOCIATED WITH POOR WOUND HEALING RESPONSES. THE PREVALENCE OF TYPE 2 DIABETES MELLITUS (T2DM) IS ESCALATING GLOBALLY. PATIENTS SUFFER FROM MULTIPLE COMPLICATIONS INCLUDING THE DEVELOPMENT OF CHRONIC WOUNDS THAT CAN LEAD TO AMPUTATION. THESE WOUNDS ARE CHARACTERISED BY AN INFLAMMATORY ENVIRONMENT INCLUDING ELEVATED TUMOUR NECROSIS FACTOR ALPHA (TNF-ALPHA). DERMAL FIBROBLASTS (DF) ARE CRITICAL FOR EFFECTIVE WOUND HEALING, SO WE SOUGHT TO ESTABLISH WHETHER THERE WERE ANY DIFFERENCES IN DF CULTURED FROM T2DM DONORS OR THOSE WITHOUT DIABETES (ND-DF). ND- AND T2DM-DF WHEN CULTURED SIMILARLY IN VITRO SECRETED COMPARABLE CONCENTRATIONS OF TNF-ALPHA. FUNCTIONALLY, PRE-TREATMENT WITH TNF-ALPHA REDUCED THE PROLIFERATION OF ND-DF AND TRANSIENTLY ALTERED ND-DF MORPHOLOGY; HOWEVER, T2DM-DF WERE RESISTANT TO THESE TNF-ALPHA INDUCED CHANGES. IN CONTRAST, TNF-ALPHA INHIBITED ND- AND T2DM-DF MIGRATION AND MATRIX METALLOPROTEASE EXPRESSION TO THE SAME DEGREE, ALTHOUGH T2DM-DF EXPRESSED SIGNIFICANTLY HIGHER LEVELS OF TISSUE INHIBITOR OF METALLOPROTEASES (TIMP)-2. FINALLY, TNF-ALPHA SIGNIFICANTLY INCREASED THE SECRETION OF PRO-INFLAMMATORY CYTOKINES (INCLUDING CCL2, CXCL1 AND SERPINE1) IN ND-DF, WHILST THIS EFFECT IN T2DM-DF WAS BLUNTED, PRESUMABLY DUE TO THE TENDENCY TO HIGHER BASELINE PRO-INFLAMMATORY CYTOKINE EXPRESSION OBSERVED IN THIS CELL TYPE. COLLECTIVELY, THESE DATA DEMONSTRATE THAT T2DM-DF EXHIBIT A SELECTIVE LOSS OF RESPONSIVENESS TO TNF-ALPHA, PARTICULARLY REGARDING PROLIFERATIVE AND SECRETORY FUNCTIONS. THIS HIGHLIGHTS IMPORTANT PHENOTYPIC CHANGES IN T2DM-DF THAT MAY EXPLAIN THE SUSCEPTIBILITY TO CHRONIC WOUNDS IN THESE PATIENTS. 2021 13 662 33 BLOOD MONOCYTE TRANSCRIPTOME AND EPIGENOME ANALYSES REVEAL LOCI ASSOCIATED WITH HUMAN ATHEROSCLEROSIS. LITTLE IS KNOWN REGARDING THE EPIGENETIC BASIS OF ATHEROSCLEROSIS. HERE WE PRESENT THE CD14+ BLOOD MONOCYTE TRANSCRIPTOME AND EPIGENOME SIGNATURES ASSOCIATED WITH HUMAN ATHEROSCLEROSIS. THE TRANSCRIPTOME SIGNATURE INCLUDES TRANSCRIPTION COACTIVATOR, ARID5B, WHICH IS KNOWN TO FORM A CHROMATIN DEREPRESSOR COMPLEX WITH A HISTONE H3K9ME2-SPECIFIC DEMETHYLASE AND PROMOTE ADIPOGENESIS AND SMOOTH MUSCLE DEVELOPMENT. ARID5B CPG (CG25953130) METHYLATION IS INVERSELY ASSOCIATED WITH BOTH ARID5B EXPRESSION AND ATHEROSCLEROSIS, CONSISTENT WITH THIS CPG RESIDING IN AN ARID5B ENHANCER REGION, BASED ON CHROMATIN CAPTURE AND HISTONE MARKS DATA. MEDIATION ANALYSIS SUPPORTS ASSUMPTIONS THAT ARID5B EXPRESSION MEDIATES EFFECTS OF CG25953130 METHYLATION AND SEVERAL CARDIOVASCULAR DISEASE RISK FACTORS ON ATHEROSCLEROTIC BURDEN. IN LIPOPOLYSACCHARIDE-STIMULATED HUMAN THP1 MONOCYTES, ARID5B KNOCKDOWN REDUCED EXPRESSION OF GENES INVOLVED IN ATHEROSCLEROSIS-RELATED INFLAMMATORY AND LIPID METABOLISM PATHWAYS, AND INHIBITED CELL MIGRATION AND PHAGOCYTOSIS. THESE DATA SUGGEST THAT ARID5B EXPRESSION, POSSIBLY REGULATED BY AN EPIGENETICALLY CONTROLLED ENHANCER, PROMOTES ATHEROSCLEROSIS BY DYSREGULATING IMMUNOMETABOLISM TOWARDS A CHRONIC INFLAMMATORY PHENOTYPE.THE MOLECULAR MECHANISMS MEDIATING THE IMPACT OF ENVIRONMENTAL FACTORS IN ATHEROSCLEROSIS ARE UNCLEAR. HERE, THE AUTHORS EXAMINE CD14+ BLOOD MONOCYTE'S TRANSCRIPTOME AND EPIGENOME SIGNATURES TO FIND DIFFERENTIAL METHYLATION AND EXPRESSION OF ARID5B TO BE ASSOCIATED WITH HUMAN ATHEROSCLEROSIS. 2017 14 222 33 ACUTE LIVER STEATOSIS TRANSLATIONALLY CONTROLS THE EPIGENETIC REGULATOR MIER1 TO PROMOTE LIVER REGENERATION IN A STUDY WITH MALE MICE. THE EARLY PHASE LIPID ACCUMULATION IS ESSENTIAL FOR LIVER REGENERATION. HOWEVER, WHETHER THIS ACUTE LIPID ACCUMULATION CAN SERVE AS SIGNALS TO DIRECT LIVER REGENERATION RATHER THAN SIMPLY PROVIDING BUILDING BLOCKS FOR CELL PROLIFERATION REMAINS UNCLEAR. THROUGH IN VIVO CRISPR SCREENING, WE IDENTIFY MIER1 (MESODERM INDUCTION EARLY RESPONSE 1) AS A KEY EPIGENETIC REGULATOR THAT BRIDGES THE ACUTE LIPID ACCUMULATION AND CELL CYCLE GENE EXPRESSION DURING LIVER REGENERATION IN MALE ANIMALS. PHYSIOLOGICALLY, LIVER ACUTE LIPID ACCUMULATION INDUCES THE PHOSPHORYLATION OF EIF2S1(EUKARYOTIC TRANSLATION INITIATION FACTOR 2), WHICH CONSEQUENTLY ATTENUATED MIER1 TRANSLATION. MIER1 DOWNREGULATION IN TURN PROMOTES CELL CYCLE GENE EXPRESSION AND REGENERATION THROUGH CHROMATIN REMODELING. IMPORTANTLY, THE LIPIDS-EIF2S1-MIER1 PATHWAY IS IMPAIRED IN ANIMALS WITH CHRONIC LIVER STEATOSIS; WHEREAS MIER1 DEPLETION SIGNIFICANTLY IMPROVES REGENERATION IN THESE ANIMALS. TAKEN TOGETHER, OUR STUDIES IDENTIFY AN EPIGENETIC MECHANISM BY WHICH THE EARLY PHASE LIPID REDISTRIBUTION FROM ADIPOSE TISSUE TO LIVER DURING REGENERATION IMPACTS HEPATOCYTE PROLIFERATION, AND SUGGEST A POTENTIAL STRATEGY TO BOOST LIVER REGENERATION. 2023 15 984 31 CHRONIC PSYCHOLOGICAL STRESS ALTERS GENE EXPRESSION IN RAT COLON EPITHELIAL CELLS PROMOTING CHROMATIN REMODELING, BARRIER DYSFUNCTION AND INFLAMMATION. CHRONIC STRESS IS COMMONLY ASSOCIATED WITH ENHANCED ABDOMINAL PAIN (VISCERAL HYPERSENSITIVITY), BUT THE CELLULAR MECHANISMS UNDERLYING HOW CHRONIC STRESS INDUCES VISCERAL HYPERSENSITIVITY ARE POORLY UNDERSTOOD. IN THIS STUDY, WE EXAMINED CHANGES IN GENE EXPRESSION IN COLON EPITHELIAL CELLS FROM A RAT MODEL USING RNA-SEQUENCING TO EXAMINE STRESS-INDUCED CHANGES TO THE TRANSCRIPTOME. FOLLOWING CHRONIC STRESS, THE MOST SIGNIFICANTLY UP-REGULATED GENES INCLUDED ATG16L1, COQ10B, DCAF13, NAT2, PTBP2, RRAS2, SPINK4 AND DOWN-REGULATED GENES INCLUDING ABAT, CITED2, CNNM2, DAB2IP, PLEKHM1, SCD2, AND TAB2. THE PRIMARY ALTERED BIOLOGICAL PROCESSES REVEALED BY NETWORK ENRICHMENT ANALYSIS WERE INFLAMMATION/IMMUNE RESPONSE, TISSUE MORPHOGENESIS AND DEVELOPMENT, AND NUCLEOSOME/CHROMATIN ASSEMBLY. THE MOST SIGNIFICANTLY DOWN-REGULATED PROCESS WAS THE DIGESTIVE SYSTEM DEVELOPMENT/FUNCTION, WHEREAS THE MOST SIGNIFICANTLY UP-REGULATED PROCESSES WERE INFLAMMATORY RESPONSE, ORGANISMAL INJURY, AND CHROMATIN REMODELING MEDIATED BY H3K9 METHYLATION. FURTHERMORE, A SUBPOPULATION OF STRESSED RATS DEMONSTRATED VERY SIGNIFICANTLY ALTERED GENE EXPRESSION AND TRANSCRIPT ISOFORMS, ENRICHED FOR THE DIFFERENTIAL EXPRESSION OF GENES INVOLVED IN THE INFLAMMATORY RESPONSE, INCLUDING UPREGULATION OF CYTOKINE AND CHEMOKINE RECEPTOR GENE EXPRESSION COUPLED WITH DOWNREGULATION OF EPITHELIAL ADHERENS AND TIGHT JUNCTION MRNAS. IN SUMMARY, THESE FINDINGS SUPPORT THAT CHRONIC STRESS IS ASSOCIATED WITH INCREASED LEVELS OF CYTOKINES AND CHEMOKINES, THEIR DOWNSTREAM SIGNALING PATHWAYS COUPLED TO DYSREGULATION OF INTESTINAL CELL DEVELOPMENT AND FUNCTION. EPIGENETIC REGULATION OF CHROMATIN REMODELING LIKELY PLAYS A PROMINENT ROLE IN THIS PROCESS. RESULTS ALSO SUGGEST THAT SUPER ENHANCERS PLAY A PRIMARY ROLE IN CHRONIC STRESS-ASSOCIATED INTESTINAL BARRIER DYSFUNCTION. 2022 16 3049 31 GENOME-WIDE ANALYSIS REVEALS ZINC TRANSPORTER ZIP9 REGULATED BY DNA METHYLATION PROMOTES RADIATION-INDUCED SKIN FIBROSIS VIA THE TGF-BETA SIGNALING PATHWAY. RADIATION-INDUCED SKIN FIBROSIS IS A DETRIMENTAL AND CHRONIC DISORDER THAT OCCURS AFTER RADIATION EXPOSURE. DNA METHYLATION HAS BEEN CHARACTERIZED AS AN IMPORTANT REGULATORY MECHANISM OF MULTIPLE PATHOLOGICAL PROCESSES. IN THIS STUDY, WE COMPARED THE GENOME-WIDE DNA METHYLATION STATUS IN RADIATION-INDUCED FIBROTIC SKIN AND ADJACENT NORMAL TISSUES OF RATS BY METHYLATED DNA IMMUNOPRECIPITATION SEQUENCING. RADIATION-INDUCED FIBROTIC SKIN SHOWED DIFFERENTIALLY METHYLATED REGIONS ASSOCIATED WITH 3,650 PROTEIN-CODING GENES, 72 MICRORNAS, 5,836 LONG NONCODING RNAS AND 3 PIWI-INTERACTING RNAS. BY INTEGRATING THE MRNA AND METHYLATION PROFILES, THE ZINC TRANSPORTER SLC39A9/ZIP9 WAS INVESTIGATED IN GREATER DETAIL. THE PROTEIN LEVEL OF ZIP9 WAS INCREASED IN IRRADIATED SKIN TISSUES OF HUMANS, MONKEYS, AND RATS, ESPECIALLY IN RADIOGENIC FIBROTIC SKIN TISSUES. RADIATION INDUCED THE DEMETHYLATION OF A CPG DINUCLEOTIDE IN EXON 1 OF ZIP9 THAT RESULTED IN RECRUITMENT OF THE TRANSCRIPTIONAL FACTOR SP1 AND INCREASED ZIP9 EXPRESSION. OVEREXPRESSION OF ZIP9 RESULTED IN ACTIVATION OF THE PROFIBROTIC TRANSFORMING GROWTH FACTOR-BETA SIGNALING PATHWAY THROUGH PROTEIN KINASE B IN HUMAN FIBROBLASTS. IN ADDITION, RADIATION-INDUCED SKIN FIBROSIS WAS ASSOCIATED WITH INCREASED ZINC ACCUMULATION. THE ZINC CHELATOR N,N,N',N'-TETRAKIS(2-PYRIDYLMETHYL)-1,2-ETHYLENEDIAMINE ABROGATED ZIP9-INDUCED ACTIVATION OF THE TRANSFORMING GROWTH FACTOR-BETA SIGNALING PATHWAY AND ATTENUATED RADIATION-INDUCED SKIN FIBROSIS IN A RAT MODEL. IN SUMMARY, OUR FINDINGS ILLUSTRATE EPIGENETIC REGULATION OF ZIP9 AND ITS CRITICAL ROLE IN PROMOTING RADIATION-INDUCED SKIN FIBROSIS. 2020 17 3128 46 GIPC-REGULATED IGFBP-3 PROMOTES HSC MIGRATION IN VITRO AND PORTAL HYPERTENSION IN VIVO THROUGH A BETA1-INTEGRIN PATHWAY. BACKGROUND & AIMS: TRANSFORMING GROWTH FACTOR (TGF-BETA)-INDUCED ACTIVATION OF QUIESCENT HEPATIC STELLATE CELLS (HSCS) AND THEIR TRANSFORMATION TO MYOFIBROBLASTS IS A KEY EVENT IN LIVER FIBROSIS AND PORTAL HYPERTENSION. GIPC (ALSO REFERRED TO AS SYNECTIN) IS A DOWNSTREAM SIGNAL ACTIVATION MOLECULE OF TGF-BETA AND OTHER RECEPTORS. IN THIS STUDY, WE SOUGHT TO IDENTIFY NOVEL GENES TARGETED BY TGF-BETA AND GIPC AND ELUCIDATE IF AND HOW THEY MAY CONTRIBUTE TO LIVER FIBROSIS. METHODS: WE PERFORMED SEQUENTIAL MESSENGER RNA SEQUENCING ANALYSIS ON TGF-BETA-STIMULATED HSCS AND THEN ON TGF-BETA-STIMULATED HSCS IN THE PRESENCE AND ABSENCE OF GIPC ALSO REFERRED TO AS SYNECTIN (GIPC) KNOCKDOWN. INSULIN-LIKE GROWTH FACTOR BINDING PROTEIN-3 (IGFBP-3) TRANSPORT PROTEIN EMERGED AS A TOP ACTIVATION TARGET OF BOTH TGF-BETA AND GIPC. QUANTITATIVE POLYMERASE CHAIN REACTION, ENZYME-LINKED IMMUNOSORBENT ASSAY, TARGETED CHROMATIN IMMUNOPRECIPITATION, AND WESTERN BLOT ANALYSIS WERE DONE FOR FURTHER CONFIRMATION. RESULTS: IGFBP-3, AN INSULIN GROWTH FACTOR TRANSPORT PROTEIN, EMERGED AS A TOP ACTIVATION TARGET OF BOTH TGF-BETA AND GIPC, WHICH WAS CONFIRMED BY QUANTITATIVE POLYMERASE CHAIN REACTION, ENZYME-LINKED IMMUNOSORBENT ASSAY, AND WESTERN BLOT ANALYSIS. TARGETED CHROMATIN IMMUNOPRECIPITATION SHOWED THAT GIPC INCREASES THE HISTONE 3 LYSINE 27 (H3K27) ACETYLATION ACTIVATING MARK AND CONCURRENTLY DECREASES THE H3K27 INHIBITORY TRIMETHYLATION (H3K27M3) MARK, PROVIDING AN EPIGENETIC CORRELATE TO THE GENE REGULATION CHANGES. IN VIVO, GLOBAL KNOCKOUT OF IGFBP-3 MICE RESULTED IN ATTENUATION OF HSC ACTIVATION MARKERS AND ATTENUATION OF PORTAL PRESSURE IN RESPONSE TO CHRONIC LIVER INJURY MODELS. ANALYSIS OF SERUM LEVELS FROM CIRRHOTIC PATIENTS ALSO SHOWED AN IGFBP-3 INCREASE OF MORE THAN 2-FOLD COMPARED WITH HEALTHY CONTROLS. FINALLY, IN VITRO MECHANISM STUDIES SHOWED THAT IGFBP-3 PROMOTES HSC MIGRATION THROUGH INTEGRIN-DEPENDENT PHOSPHORYLATION OF PROTEIN KINASE B. CONCLUSIONS: TGF-BETA UP-REGULATES IGFBP-3 THROUGH GIPC, LEADING TO INCREASED HSC MIGRATION IN VITRO AND PROMOTES PORTAL HYPERTENSION IN VIVO. THESE STUDIES SUPPORT THE ROLE OF IGFBP-3 AS A POTENTIAL PATHOPHYSIOLOGIC TARGET OR BIOMARKER IN CHRONIC LIVER DISEASE. 2020 18 3306 37 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 19 669 32 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 20 1461 35 DISRUPTION OF RCAN1.4 EXPRESSION MEDIATED BY YY1/HDAC2 MODULATES CHRONIC RENAL ALLOGRAFT INTERSTITIAL FIBROSIS. CHRONIC ALLOGRAFT DYSFUNCTION (CAD) IS A MAJOR FACTOR THAT HINDERS KIDNEY TRANSPLANT SURVIVAL IN THE LONG RUN. EPITHELIAL-MESENCHYMAL TRANSITION (EMT) HAS BEEN CONFIRMED TO SIGNIFICANTLY CONTRIBUTE TO INTERSTITIAL FIBROSIS/TUBULAR ATROPHY (IF/TA), WHICH IS THE MAIN HISTOPATHOLOGICAL FEATURE OF CAD. ABERRANT EXPRESSION OF THE REGULATOR OF CALCINEURIN 1 (RCAN1), RECOGNIZED AS AN ENDOGENOUS INHIBITOR OF THE CALCINEURIN PHOSPHATASE, HAS BEEN SHOWN TO BE EXTENSIVELY INVOLVED IN VARIOUS KIDNEY DISEASES. HOWEVER, IT REMAINS UNCLEAR HOW RCAN1.4 REGULATES IF/TA FORMATION IN CAD PATIENTS. HEREIN, AN IN VIVO MOUSE RENAL TRANSPLANTATION MODEL AND AN IN VITRO MODEL OF HUMAN RENAL TUBULAR EPITHELIAL CELLS (HK-2) TREATED WITH TUMOR NECROSIS FACTOR-ALPHA (TNF-ALPHA) WERE EMPLOYED. OUR RESULTS PROVED THAT RCAN1.4 EXPRESSION WAS DECREASED IN VIVO AND IN VITRO, IN ADDITION TO THE UP-REGULATION OF YIN YANG 1 (YY1), A TRANSCRIPTION FACTOR THAT HAS BEEN REPORTED TO CONVEY MULTIPLE FUNCTIONS IN CHRONIC KIDNEY DISEASE (CKD). KNOCKING IN OF RCAN1.4 EFFICIENTLY ATTENUATED CHRONIC RENAL ALLOGRAFT INTERSTITIAL FIBROSIS IN VIVO AND INHIBITED TNF-ALPHA-INDUCED EMT IN VITRO THROUGH REGULATING ANTI-OXIDATIVE STRESS AND THE CALCINEURIN/NUCLEAR FACTOR OF ACTIVATED T CELLS CYTOPLASMIC 1 (NFATC1) SIGNALING PATHWAY. IN ADDITION, SUPPRESSION OF YY1 MEDIATED BY SHRNA OR SIRNA ALLEVIATED TNF-ALPHA-INDUCED EMT THROUGH ABOLISHING REACTIVE SPECIES PARTLY IN AN RCAN1.4-DEPENDENT MANNER. NOTABLY, WE CONFIRMED THAT YY1 NEGATIVELY REGULATED RCAN1.4 TRANSCRIPTION BY DIRECTLY INTERACTING WITH THE RCAN1.4 PROMOTER. IN ADDITION, HISTONE DEACETYLASE 2 (HDAC2) INTERACTED WITH YY1 TO FORM A MULTI-MOLECULAR COMPLEX, WHICH WAS INVOLVED IN TNF-ALPHA-INDUCED RCAN1.4 TRANSCRIPTIONAL REPRESSION. THEREFORE, RCAN1.4 IS SUGGESTED TO BE MODULATED BY THE YY1/HDAC2 TRANSCRIPTION REPRESSOR COMPLEX IN AN EPIGENETIC MANNER, WHICH IS A MEDIATED NEPHROPROTECTIVE EFFECT PARTLY THROUGH MODULATING O2?- GENERATION AND THE CALCINEURIN/NFATC1 SIGNALING PATHWAY. THUS, THE YY1-RCAN1.4 AXIS CONSTITUTES AN INNOVATIVE TARGET FOR IF/TA TREATMENT IN CAD PATIENTS. 2023