1 3237 160 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 2 5332 38 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 3 3128 45 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 4 5604 41 ROSAVIN AMELIORATES HEPATIC INFLAMMATION AND FIBROSIS IN THE NASH RAT MODEL VIA TARGETING HEPATIC CELL DEATH. BACKGROUND: NON-ALCOHOLIC FATTY LIVER DISEASE (NAFLD) REPRESENTS THE MOST COMMON FORM OF CHRONIC LIVER DISEASE THAT URGENTLY NEEDS EFFECTIVE THERAPY. ROSAVIN, A MAJOR CONSTITUENT OF THE RHODIOLA ROSEA PLANT OF THE FAMILY CRASSULACEAE, IS BELIEVED TO EXHIBIT MULTIPLE PHARMACOLOGICAL EFFECTS ON DIVERSE DISEASES. HOWEVER, ITS EFFECT ON NON-ALCOHOLIC STEATOHEPATITIS (NASH), THE PROGRESSIVE FORM OF NAFLD, AND THE UNDERLYING MECHANISMS ARE NOT FULLY ILLUSTRATED. AIM: INVESTIGATE THE PHARMACOLOGICAL ACTIVITY AND POTENTIAL MECHANISM OF ROSAVIN TREATMENT ON NASH MANAGEMENT VIA TARGETING HEPATIC CELL DEATH-RELATED (HSPD1/TNF/MMP14/ITGB1) MRNAS AND THEIR UPSTREAM NONCODING RNA REGULATORS (MIRNA-6881-5P AND LNC-SPARCL1-1:2) IN NASH RATS. RESULTS: HIGH SUCROSE HIGH FAT (HSHF) DIET-INDUCED NASH RATS WERE TREATED WITH DIFFERENT CONCENTRATIONS OF ROSAVIN (10, 20, AND 30 MG/KG/DAY) FOR THE LAST FOUR WEEKS OF DIETARY MANIPULATION. THE DATA REVEALED THAT ROSAVIN HAD THE ABILITY TO MODULATE THE EXPRESSION OF THE HEPATIC CELL DEATH-RELATED RNA PANEL THROUGH THE UPREGULATION OF BOTH (HSPD1/TNF/MMP14/ITGB1) MRNAS AND THEIR EPIGENETIC REGULATORS (MIRNA-6881-5P AND LNC-SPARCL1-1:2). MOREOVER, ROSAVIN AMELIORATED THE DETERIORATION IN BOTH LIVER FUNCTIONS AND LIPID PROFILE, AND THEREBY IMPROVED THE HEPATIC INFLAMMATION, FIBROSIS, AND APOPTOSIS, AS EVIDENCED BY THE DECREASED PROTEIN LEVELS OF IL6, TNF-ALPHA, AND CASPASE-3 IN LIVER SECTIONS OF TREATED ANIMALS COMPARED TO THE UNTREATED NASH RATS. CONCLUSION: ROSAVIN HAS DEMONSTRATED A POTENTIAL ABILITY TO ATTENUATE DISEASE PROGRESSION AND INHIBIT HEPATIC CELL DEATH IN THE NASH ANIMAL MODEL. THE PRODUCED EFFECT WAS CORRELATED WITH UPREGULATION OF THE HEPATIC CELL DEATH-RELATED (HSPD1, TNF, MMP14, AND ITGB1) MRNAS-(MIRNA-6881-5P-(LNC-SPARCL1-1:2) RNA PANEL. 2022 5 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 6 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 7 4400 33 MODULATION OF HEPATIC STELLATE CELLS BY MUTAFLOR((R)) PROBIOTIC IN NON-ALCOHOLIC FATTY LIVER DISEASE MANAGEMENT. BACKGROUND: NAFLD AND NASH ARE EMERGING AS PRIMARY CAUSES OF CHRONIC LIVER DISEASE, INDICATING A NEED FOR AN EFFECTIVE TREATMENT. MUTAFLOR(R) PROBIOTIC, A MICROBIAL TREATMENT OF INTEREST, WAS EFFECTIVE IN SUSTAINING REMISSION IN ULCERATIVE COLITIS PATIENTS. OBJECTIVE: TO CONSTRUCT A GENETIC-EPIGENETIC NETWORK LINKED TO HSC SIGNALING AS A MODULATOR OF NAFLD/NASH PATHOGENESIS, THEN ASSESS THE EFFECTS OF MUTAFLOR((R)) ON THIS NETWORK. METHODS: FIRST, IN SILICO ANALYSIS WAS USED TO CONSTRUCT A GENETIC-EPIGENETIC NETWORK LINKED TO HSC SIGNALING. SECOND, AN INVESTIGATION USING RATS, INCLUDING HFHSD INDUCED NASH AND MUTAFLOR((R)) TREATED ANIMALS, WAS DESIGNED. EXPERIMENTAL PROCEDURES INCLUDED BIOCHEMICAL AND HISTOPATHOLOGIC ANALYSIS OF RAT BLOOD AND LIVER SAMPLES. AT THE MOLECULAR LEVEL, THE EXPRESSION OF GENETIC (FOXA2, TEAD2, AND LATS2 MRNAS) AND EPIGENETIC (MIR-650, RPARP AS-1 LNCRNA) NETWORK WAS MEASURED BY REAL-TIME PCR. PCR RESULTS WERE VALIDATED WITH IMMUNOHISTOCHEMISTRY (ALPHA-SMA AND LATS2). TARGET EFFECTOR PROTEINS, IL-6 AND TGF-BETA, WERE ESTIMATED BY ELISA. RESULTS: MUTAFLOR((R)) ADMINISTRATION MINIMIZED BIOCHEMICAL AND HISTOPATHOLOGIC ALTERATIONS CAUSED BY NAFLD/NASH. HSC ACTIVATION AND EXPRESSION OF PROFIBROGENIC IL-6 AND TGF-BETA EFFECTOR PROTEINS WERE REDUCED VIA INHIBITION OF HEDGEHOG AND HIPPO PATHWAYS. PATHWAYS MAY HAVE BEEN INHIBITED THROUGH UPREGULATION OF RPARP AS-1 LNCRNA WHICH IN TURN DOWNREGULATED THE EXPRESSION OF MIR-650, FOXA2 MRNA AND TEAD2 MRNA AND UPREGULATED LATS2 MRNA EXPRESSION. CONCLUSION: MUTAFLOR((R)) MAY SLOW THE PROGRESSION OF NAFLD/NASH BY MODULATING A GENETIC-EPIGENETIC NETWORK LINKED TO HSC SIGNALING. THE PROBIOTIC MAY BE A USEFUL MODALITY FOR THE PREVENTION AND TREATMENT OF NAFLD/NASH. 2022 8 1335 29 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 9 6385 44 THE ROLE OF PROMYELOCYTIC LEUKEMIA PROTEIN IN STEATOSIS-ASSOCIATED HEPATIC TUMORS RELATED TO CHRONIC HEPATITIS B VIRUS INFECTION. THE PERSISTENCE OF HEPATITIS B SURFACE ANTIGEN (HBSAG) IS A RISK FACTOR FOR THE DEVELOPMENT OF STEATOSIS-ASSOCIATED TUMORS IN CHRONIC HEPATITIS B VIRUS (HBV) INFECTION, YET LITTLE IS KNOWN ABOUT THE METABOLIC LINK WITH THIS FACTOR. WE CORRELATED HBV-RELATED PATHOGENESIS IN GENETICALLY ENGINEERED MICE AND HUMAN CARRIERS WITH METABOLIC PROTEOMICS AND LIPOGENIC GENE EXPRESSION PROFILES. THE IMMUNOHISTOCHEMISTRY SHOWED THAT THE PROMYELOCYTIC LEUKEMIA PROTEIN (PML, A TUMOR SUPPRESSOR INVOLVED IN GENOME MAINTENANCE AND FATTY ACID OXIDATION), BEING INVERSELY INFLUENCED BY THE DYNAMIC HBSAG LEVELS FROM ACUTE PHASE TO SEROCLEARANCE, APPEARED AS A LIPO-METABOLIC SWITCH LINKING HBSAG-INDUCED STEATOSIS (LIPOGENESIS) TO HBSAG-LOST FAT-BURNING HEPATOCARCINOGENESIS (LIPOLYSIS). KNOCKDOWN OF PML IN HBSAG-TRANSGENIC MICE PREDISPOSED TO OBESITY AND DROVE EARLY STEATOSIS-SPECIFIC LIVER TUMORIGENESIS. PROTEOME ANALYSIS REVEALED THAT THE SIGNALING PATHWAYS CORRESPONDING TO ENERGY METABOLISM AND ITS REGULATORS WERE FREQUENTLY ALTERED BY SUPPRESSION OR DEPLETION OF PML IN THE HBSAG-TRANSGENIC MICE, MAINLY INCLUDING OXIDATIVE PHOSPHORYLATION AND FATTY ACID METABOLISM. EXPRESSION PROFILING FURTHER IDENTIFIED UPREGULATION OF STEAROYL-COA DESATURASE 1 (SCD1) AND EPIGENETIC METHYLATION OF NDUFA13 IN THE MITOCHONDRIAL RESPIRATORY CHAIN AND THE CELL CYCLE INHIBITOR CDKN1C IN CONCORDANCE TO THE INCREASED SEVERITY OF LIPODYSTROPHY AND NEOPLASIA IN THE LIVERS OF HBSAG-TRANSGENIC MICE WITH PML INSUFFICIENCY. THE DEFECT IN LIPOLYSIS IN PML-DEFICIENT HBSAG-TRANSGENIC MICE MADE THE HBSAG-INDUCED ADIPOSE-LIKE LIVER TUMORS VULNERABLE TO SYNTHETIC LETHALITY FROM TOXIC SATURATED FAT ACCUMULATION WITH A SCD1 INHIBITOR. OUR FINDINGS PROVIDE MECHANISTIC INSIGHTS INTO THE EVOLUTION OF STEATOSIS-ASSOCIATED HEPATIC TUMORS DRIVEN BY RECIPROCAL INTERACTIONS OF HBSAG AND PML, AND A POTENTIAL UTILITY OF LIPID METABOLIC REPROGRAMMING AS A TREATMENT TARGET. 2018 10 316 37 ALCOHOL-ASSOCIATED FOLATE DISTURBANCES RESULT IN ALTERED METHYLATION OF FOLATE-REGULATING GENES. FOLATE PLAYS A CRITICAL ROLE IN MAINTAINING NORMAL METABOLIC, ENERGY, DIFFERENTIATION AND GROWTH STATUS OF ALL MAMMALIAN CELLS. THE STEADY-STATE ACCUMULATION OF FOLATE SEEMS TO DEPEND ON THE ACTIVITY OF TWO ENZYMES: FOLYLPOLYGLUTAMATE SYNTHETASE (FPGS), WHICH ADDS GLUTAMATE RESIDUES, AND GAMMA-GLUTAMYL HYDROLASE (GGH), WHICH REMOVES THEM, ENABLING IT TO BE TRANSPORTED ACROSS THE BIOLOGICAL MEMBRANES. OVEREXPRESSION OF GGH AND DOWNREGULATION OF FPGS WOULD BE EXPECTED TO DECREASE INTRACELLULAR FOLATE IN ITS POLYGLUTAMYLATED FORM, THEREBY INCREASING EFFLUX OF FOLATE AND ITS RELATED MOLECULES, WHICH MIGHT LEAD TO RESISTANCE TO DRUGS OR FOLATE DEFICIENCY. THE STUDY WAS SOUGHT TO DELINEATE THE ACTIVITY OF GGH AND EXPRESSION FPGS IN TISSUES INVOLVED IN FOLATE HOMEOSTASIS DURING ALCOHOLISM AND THE EPIGENETIC REGULATION OF THESE ENZYMES AND TRANSPORTERS REGULATING INTRACELLULAR FOLATE LEVELS. WE DETERMINED THE ACTIVITY OF GGH AND EXPRESSION OF FPGS IN TISSUES AFTER 3 MONTHS OF ETHANOL FEEDING TO RATS AT 1 G/KG BODY WEIGHT/DAY. THE RESULTS SHOWED THAT THERE WAS NOT ANY SIGNIFICANT CHANGE IN THE ACTIVITY OF FOLATE HYDROLYZING ENZYME GGH IN ETHANOL-FED RATS WHILE THERE WAS SIGNIFICANT DOWN REGULATION IN THE EXPRESSION OF FPGS. ETHANOL FEEDING DECREASED THE TOTAL AS WELL AS POLYGLUTAMATED FOLATE LEVELS. THERE WAS TISSUE-SPECIFIC HYPER/HYPO METHYLATION OF FOLATE TRANSPORTER GENES VIZ. PCFT AND RFC BY CHRONIC ETHANOL FEEDING. MOREOVER, HYPERMETHYLATION OF FPGS GENE WAS OBSERVED IN INTESTINE AND KIDNEY WITHOUT ANY CHANGE IN METHYLATION LEVELS OF GGH IN THE ETHANOL-FED RATS. IN CONCLUSION, THE INITIAL DECONJUGATION OF POLYGLUTAMYLATED FOLATE BY GGH WAS NOT IMPAIRED IN ETHANOL-FED RATS WHILE THE CONVERSION OF MONOGLUTAMYLATED FOLATE TO POLYGLUTAMYLATED FORM MIGHT BE IMPAIRED. THERE WAS TISSUE-SPECIFIC ALTERED METHYLATION OF FOLATE TRANSPORTER GENES BY CHRONIC ETHANOL FEEDING. 2012 11 5636 39 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 12 984 35 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 13 2758 25 EXPRESSION OF HORMONAL CARCINOGENESIS GENES AND RELATED REGULATORY MICRORNAS IN UTERUS AND OVARIES OF DDT-TREATED FEMALE RATS. THE INSECTICIDE DICHLORODIPHENYLTRICHLOROETHANE (DDT) IS A NONMUTAGENIC XENOBIOTIC COMPOUND ABLE TO EXERT ESTROGEN-LIKE EFFECTS RESULTING IN ACTIVATION OF ESTROGEN RECEPTOR-ALPHA (ERALPHA) FOLLOWED BY CHANGED EXPRESSION OF ITS DOWNSTREAM TARGET GENES. IN ADDITION, STUDIES PERFORMED OVER RECENT YEARS SUGGEST THAT DDT MAY ALSO INFLUENCE EXPRESSION OF MICRORNAS. HOWEVER, AN IMPACT OF DDT ON EXPRESSION OF ER, MICRORNAS, AND RELATED TARGET GENES HAS NOT BEEN FULLY ELUCIDATED. HERE, USING REAL-TIME PCR, WE ASSESSED CHANGES IN EXPRESSION OF KEY GENES INVOLVED IN HORMONAL CARCINOGENESIS AS WELL AS POTENTIALLY RELATED REGULATORY ONCOGENIC/TUMOR SUPPRESSOR MICRORNAS AND THEIR TARGET GENES IN THE UTERUS AND OVARIES OF FEMALE WISTAR RATS DURING SINGLE AND CHRONIC MULTIPLE-DOSE DDT EXPOSURE. WE FOUND THAT APPLYING DDT RESULTS IN ALTERED EXPRESSION OF MICRORNAS-221, -222, -205, -126A, AND -429, THEIR TARGET GENES (PTEN, DICER1), AS WELL AS GENES INVOLVED IN HORMONAL CARCINOGENESIS (ESR1, PGR, CCND1, CYP19A1). NOTABLY, CYP19A1 EXPRESSION SEEMS TO BE ALSO REGULATED BY MICRORNAS-221, -222, AND -205. THE DATA SUGGEST THAT EPIGENETIC EFFECTS INDUCED BY DDT AS A POTENTIAL CARCINOGEN MAY BE BASED ON AT LEAST TWO MECHANISMS: (I) ACTIVATION OF ERALPHA FOLLOWED BY ALTERED EXPRESSION OF THE TARGET GENES ENCODING RECEPTOR PGR AND CCND1 AS WELL AS IMPAIRED EXPRESSION OF CYP19A1, AFFECTING, THEREBY, CELL HORMONE BALANCE; AND (II) CHANGED EXPRESSION OF MICRORNAS RESULTING IN IMPAIRED EXPRESSION OF RELATED TARGET GENES INCLUDING REDUCED LEVEL OF CYP19A1 MRNA. 2017 14 3390 29 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 15 4360 32 MIR-6769B-5P TARGETS CCND-1 TO REGULATE PROLIFERATION IN CADMIUM-TREATED PLACENTAL TROPHOBLASTS: ASSOCIATION WITH THE IMPAIRMENT OF FETAL GROWTH. ENVIRONMENTAL CADMIUM (CD) IS POSITIVELY ASSOCIATED WITH PLACENTAL IMPAIRMENT AND FETAL GROWTH RETARDATION. NEVERTHELESS, ITS POTENTIAL MECHANISMS REMAIN UNCLEAR. MICRORNAS (MIRNAS) ARE KNOWN TO INFLUENCE PLACENTAL DEVELOPMENT AND FETAL GROWTH. THIS WORK WAS AIMED TO DETERMINE WHICH MIRNAS ARE INVOLVED IN CD-IMPAIRED PLACENTAL AND FETAL DEVELOPMENT BASED ON THE MRNA AND MIRNA EXPRESSION PROFILES ANALYSIS. AS A RESULT, GESTATIONAL CD EXPOSURE DECEASED FETAL AND PLACENTAL WEIGHT, AND REDUCED THE PROTEIN LEVEL OF PCNA IN HUMAN AND MOUSE PLACENTAE. FURTHERMORE, THE RESULTS OF MRNA MICROARRAY SHOWED THAT CD-DOWNREGULATED MRNAS WERE PREDICTIVELY CORRELATED WITH SEVERAL BIOLOGICAL PROCESSES, INCLUDING CELL PROLIFERATION, DIFFERENTIATION AND MOTILITY. IN ADDITION, THE RESULTS OF MIRNA MICROARRAY AND QPCR ASSAY DEMONSTRATED THAT CD SIGNIFICANTLY INCREASED THE LEVEL OF MIR-6769B-5P, MIR-146B-5P AND MIR-452-5P. INTEGRATED ANALYSIS OF CD-UPREGULATED MIRNAS PREDICTED TARGET GENES AND CD-DOWNREGULATED MRNAS FOUND THAT OVERLAPPING MRNAS, SUCH AS CCND1, CDK13, RINT1 AND CDC26 WERE ALSO SIGNIFICANTLY ASSOCIATED WITH CELL PROLIFERATION. FURTHER EXPERIMENTS SHOWED THAT MIR-6769B-5P INHIBITOR, BUT NOT MIR-146B-5P AND MIR-452-5P, MARKEDLY REVERSED CD-DOWNREGULATED THE EXPRESSION OF PROLIFERATION-RELATED MRNAS, AND THEREBY RESTORED CD-DECREASED THE PROTEINS LEVEL OF CCND1 AND PCNA IN HUMAN PLACENTAL TROPHOBLASTS. DUAL LUCIFERASE REPORTER ASSAY FURTHER REVEALED THAT MIR-6769B-5P DIRECTLY TARGETS CCND1. FINALLY, THE CASE-CONTROL STUDY DEMONSTRATED THAT INCREASED MIR-6769B-5P LEVEL AND IMPAIRED CELL PROLIFERATION WERE OBSERVED IN SMALL-FOR-GESTATIONAL-AGE HUMAN PLACENTAE. IN CONCLUSION, MIR-6769B-5P TARGETS CCND-1 TO REGULATE PROLIFERATION IN CD-TREATED PLACENTAL TROPHOBLASTS, WHICH IS ASSOCIATED WITH THE IMPAIRMENT OF FETAL GROWTH. OUR FINDINGS IMPLY THAT PLACENTAL MIR-6769B-5P MAY BE USED AS AN EPIGENETIC MARKER FOR ENVIRONMENTAL POLLUTANTS-CAUSED FETAL GROWTH RESTRICTION AND ITS LATE-ONSET CHRONIC DISEASES. 2021 16 2427 34 EPIGENETIC SILENCING OF MICRORNA-125B-5P PROMOTES LIVER FIBROSIS IN NONALCOHOLIC FATTY LIVER DISEASE VIA INTEGRIN ALPHA8-MEDIATED ACTIVATION OF RHOA SIGNALING PATHWAY. BACKGROUND: NONALCOHOLIC FATTY LIVER DISEASE (NAFLD) IS ONE OF THE MOST COMMON CHRONIC LIVER DISEASES THAT MAY PROGRESS TO LIVER FIBROSIS OR CANCER. THE PRESENT STUDY AIMED TO INVESTIGATE THE ROLE OF MICRORNA-125B-5P (MIR-125B-5P) IN NAFLD AND TO FURTHER EXPLORE UNDERLYING MOLECULAR MECHANISMS. METHODS: A MOUSE MODEL OF NAFLD WAS CONSTRUCTED BY HIGH CHOLESTEROL DIET FEEDING AND A CELL-MODEL WAS DEVELOPED BY TREATING THE MOUSE LIVER CELL LINE NCTC1469 WITH PALMITIC ACID. GAIN- AND LOSS-OF-FUNCTION EXPERIMENTS WERE PERFORMED TO DETERMINE THE EFFECTS OF MIR-125B-5P, INTEGRIN ALPHA8 (ITGA8), AND THE RHOA SIGNALING PATHWAY ON LIVER FIBROSIS IN NAFLD. AFTER THE EXPRESSION LEVELS OF MIR-125B-5P, ITGA8, AND RHOA WERE DETERMINED, LIVER FIBROSIS WAS EVALUATED IN VIVO AND IN VITRO. THE BINDING RELATIONSHIP OF MIR-125B-5P AND ITGA8 WAS THEN VALIDATED. FINALLY, MIR-125B-5P PROMOTER METHYLATION IN NAFLD LIVER TISSUES AND CELLS WAS DETERMINED. RESULTS: IN NAFLD CLINICAL SAMPLES, MOUSE MODEL, AND CELL-MODEL, MIR-125B-5P EXPRESSION WAS REDUCED, WHILE ITGA8 EXPRESSION WAS INCREASED. MOREOVER, MIR-125B-5P TARGETED AND DOWNREGULATED ITGA8, LEADING TO INHIBITION OF THE RHOA SIGNALING PATHWAY. IN NAFLD LIVER TISSUES AND CELLS, THE CPG ISLAND IN THE MIR-125B-5P PROMOTER WAS METHYLATED, CAUSING EPIGENETIC SILENCING OF MIR-125B-5P. BOTH MIR-125B-5P SILENCING AND ITGA8 OVEREXPRESSION PROMOTED IN VITRO AND IN VIVO LIVER FIBROSIS IN NAFLD VIA ACTIVATION OF THE RHOA SIGNALING PATHWAY. CONCLUSIONS: COLLECTIVELY, EPIGENETIC SILENCING OF MIR-125B-5P UPREGULATES ITGA8 EXPRESSION TO ACTIVATE THE RHOA SIGNALING PATHWAY, LEADING TO LIVER FIBROSIS IN NAFLD. 2020 17 3096 39 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 18 4001 44 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 19 222 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 20 1429 39 DIFFERENTIAL EXPRESSION OF MICRORNAS IN THE HIPPOCAMPI OF MALE AND FEMALE RODENTS AFTER CHRONIC ALCOHOL ADMINISTRATION. BACKGROUND: WOMEN ARE MORE VULNERABLE THAN MEN TO THE NEUROTOXICITY AND SEVERE BRAIN DAMAGE CAUSED BY CHRONIC HEAVY ALCOHOL USE. IN ADDITION, BRAIN DAMAGE DUE TO CHRONIC HEAVY ALCOHOL USE MAY BE ASSOCIATED WITH SEX-DEPENDENT EPIGENETIC MODIFICATIONS. THIS STUDY AIMED TO IDENTIFY MICRORNAS (MIRNAS) AND THEIR TARGET GENES THAT ARE DIFFERENTIALLY EXPRESSED IN THE HIPPOCAMPI OF MALE AND FEMALE ANIMAL MODELS IN RESPONSE TO ALCOHOL. METHODS: AFTER CHRONIC ALCOHOL ADMINISTRATION (3~3.5 G/KG/DAY) IN MALE (CONTROL, N = 10; ALCOHOL, N = 12) OR FEMALE (CONTROL, N = 10; ALCOHOL, N = 12) SPRAGUE-DAWLEY RATS FOR 6 WEEKS, WE MEASURED BODY WEIGHTS AND DOUBLECORTIN (DCX; A NEUROGENESIS MARKER) CONCENTRATIONS AND ANALYZED UP- OR DOWNREGULATED MIRNAS USING GENECHIP MIRNA 4.0 ARRAYS. THE DIFFERENTIALLY EXPRESSED MIRNAS AND THEIR PUTATIVE TARGET GENES WERE VALIDATED BY RT-QPCR. RESULTS: ALCOHOL ATTENUATED BODY WEIGHT GAIN ONLY IN THE MALE GROUP. ON THE OTHER HAND, ALCOHOL LED TO INCREASED SERUM AST IN FEMALE RATS AND DECREASED SERUM TOTAL CHOLESTEROL CONCENTRATIONS IN MALE RATS. THE EXPRESSION OF DCX WAS SIGNIFICANTLY REDUCED IN THE HIPPOCAMPI OF MALE ALCOHOL-TREATED RATS. NINE MIRNAS WERE SIGNIFICANTLY UP- OR DOWNREGULATED IN MALE ALCOHOL-TREATED RATS, INCLUDING UPREGULATION OF MIR-125A-3P, LET-7A-5P, AND MIR-3541, AND DOWNREGULATION OF THEIR TARGET GENES (PRDM5, SUV39H1, PTPRZ1, MAPK9, ING4, WT1, NKX3-1, DAB2IP, RNF152, RIPK1, LIN28A, APBB3, NRAS, AND ACVR1C). ON THE OTHER HAND, 7 MIRNAS WERE SIGNIFICANTLY UP- OR DOWNREGULATED IN ALCOHOL-TREATED FEMALE RATS, INCLUDING DOWNREGULATION OF MIR-881-3P AND MIR-504 AND UPREGULATION OF THEIR TARGET GENES (NAA50, CLOCK, CBFB, ARIH1, UBE2G1, AND GNG7). CONCLUSIONS: THESE RESULTS SUGGEST THAT CHRONIC HEAVY ALCOHOL USE PRODUCES SEX-DEPENDENT EFFECTS ON NEUROGENESIS AND MIRNA EXPRESSION IN THE HIPPOCAMPUS AND THAT SEX DIFFERENCES SHOULD BE CONSIDERED WHEN DEVELOPING MIRNA BIOMARKERS TO DIAGNOSE OR TREAT ALCOHOLICS. 2020