1 1022 135 CIRCULAR RNA HSA_CIRC_0098181 INHIBITS METASTASIS IN HEPATOCELLULAR CARCINOMA BY ACTIVATING THE HIPPO SIGNALING PATHWAY VIA INTERACTION WITH EEF2. INTRODUCTION AND OBJECTIVES: THE DEVELOPMENT OF HEPATOCELLULAR CARCINOMA (HCC) IS A MULTI-STEP PROCESS THAT ACCUMULATES GENETIC AND EPIGENETIC ALTERATIONS, INCLUDING CHANGES IN CIRCULAR RNA (CIRCRNA). THIS STUDY AIMED TO UNDERSTAND THE ALTERATIONS IN CIRCRNA EXPRESSION IN HCC DEVELOPMENT AND METASTASIS AND TO EXPLORE THE BIOLOGICAL FUNCTIONS OF CIRCRNA. MATERIALS AND METHODS: TEN PAIRS OF ADJACENT CHRONIC HEPATITIS TISSUES AND HCC TISSUES FROM PATIENTS WITHOUT VENOUS METASTASES, AND TEN HCC TISSUES FROM PATIENTS WITH VENOUS METASTASES WERE ANALYZED USING HUMAN CIRCRNA MICROARRAYS. DIFFERENTIALLY EXPRESSED CIRCRNAS WERE THEN VALIDATED BY QUANTITATIVE REAL-TIME PCR. IN VITRO AND IN VIVO ASSAYS WERE PERFORMED TO ASSESS THE ROLES OF THE CIRCRNA IN HCC PROGRESSION. RNA PULL-DOWN ASSAY, MASS SPECTROMETRY ANALYSIS, AND RNA-BINDING PROTEIN IMMUNOPRECIPITATION WERE CONDUCTED TO EXPLORE THE PROTEIN PARTNERS OF THE CIRCRNA. RESULTS: CIRCRNA MICROARRAYS REVEALED THAT THE EXPRESSION PATTERNS OF CIRCRNAS ACROSS THE THREE GROUPS WERE SIGNIFICANTLY DIFFERENT. AMONG THESE, HSA_CIRC_0098181 WAS VALIDATED TO BE LOWLY EXPRESSED AND ASSOCIATED WITH POOR PROGNOSIS IN HCC PATIENTS. ECTOPIC EXPRESSION OF HSA_CIRC_0098181 DELAYED HCC METASTASIS IN VITRO AND IN VIVO. MECHANISTICALLY, HSA_CIRC_0098181 SEQUESTERED EUKARYOTIC TRANSLATION ELONGATION FACTOR 2 (EEF2) AND DISSOCIATED EEF2 FROM FILAMENTOUS ACTIN (F-ACTIN) TO PREVENT F-ACTIN FORMATION, WHICH BLOCKED ACTIVATION OF THE HIPPO SIGNALING PATHWAY. IN ADDITION, THE RNA BINDING PROTEIN QUAKING-5 BOUND DIRECTLY TO HSA_CIRC_0098181 AND INDUCED ITS BIOGENESIS. CONCLUSIONS: OUR STUDY REVEALS CHANGES IN CIRCRNA EXPRESSION FROM CHRONIC HEPATITIS, PRIMARY HCC, TO METASTATIC HCC. FURTHER, THE QKI5-HSA_CIRC_0098181-EEF2-HIPPO SIGNALING PATHWAY EXERTS A REGULATORY ROLE IN HCC. 2023 2 5939 33 TARGETING MECHANOTRANSDUCTION AT THE TRANSCRIPTIONAL LEVEL: YAP AND BRD4 ARE NOVEL THERAPEUTIC TARGETS FOR THE REVERSAL OF LIVER FIBROSIS. LIVER FIBROSIS IS THE RESULT OF A DEREGULATED WOUND HEALING PROCESS CHARACTERIZED BY THE EXCESSIVE DEPOSITION OF EXTRACELLULAR MATRIX. HEPATIC STELLATE CELLS (HSCS), WHICH ARE ACTIVATED IN RESPONSE TO LIVER INJURY, ARE THE MAJOR SOURCE OF EXTRACELLULAR MATRIX AND DRIVE THE WOUND HEALING PROCESS. HOWEVER, CHRONIC LIVER DAMAGE LEADS TO PERPETUAL HSC ACTIVATION, PROGRESSIVE FORMATION OF PATHOLOGICAL SCAR TISSUE AND ULTIMATELY, CIRRHOSIS AND ORGAN FAILURE. HSC ACTIVATION IS TRIGGERED LARGELY IN RESPONSE TO MECHANOSIGNALING FROM THE MICROENVIRONMENT, WHICH INDUCES A PROFIBROTIC NUCLEAR TRANSCRIPTION PROGRAM THAT PROMOTES HSC PROLIFERATION AND EXTRACELLULAR MATRIX SECRETION THEREBY SETTING UP A POSITIVE FEEDBACK LOOP LEADING TO MATRIX STIFFENING AND SELF-SUSTAINED, PATHOLOGICAL, HSC ACTIVATION. DESPITE THE SIGNIFICANT PROGRESS IN OUR UNDERSTANDING OF LIVER FIBROSIS, THE MOLECULAR MECHANISMS THROUGH WHICH THE EXTRACELLULAR MATRIX PROMOTES HSC ACTIVATION ARE NOT WELL UNDERSTOOD AND NO EFFECTIVE THERAPIES HAVE BEEN APPROVED TO DATE THAT CAN TARGET THIS EARLY, REVERSIBLE, STAGE IN LIVER FIBROSIS. SEVERAL NEW LINES OF INVESTIGATION NOW PROVIDE IMPORTANT INSIGHT INTO THIS AREA OF STUDY AND IDENTIFY TWO NUCLEAR TARGETS WHOSE INHIBITION HAS THE POTENTIAL OF REVERSING LIVER FIBROSIS BY INTERFERING WITH HSC ACTIVATION: YES-ASSOCIATED PROTEIN (YAP), A TRANSCRIPTIONAL CO-ACTIVATOR AND EFFECTOR OF THE MECHANOSENSITIVE HIPPO PATHWAY, AND BROMODOMAIN-CONTAINING PROTEIN 4 (BRD4), AN EPIGENETIC REGULATOR OF GENE EXPRESSION. YAP AND BRD4 ACTIVITY IS INDUCED IN RESPONSE TO MECHANICAL STIMULATION OF HSCS AND EACH PROTEIN INDEPENDENTLY CONTROLS WAVES OF EARLY GENE EXPRESSION NECESSARY FOR HSC ACTIVATION. SIGNIFICANTLY, INHIBITION OF EITHER PROTEIN CAN REVERT THE CHRONIC ACTIVATION OF HSCS AND IMPEDE PATHOLOGICAL PROGRESSION OF LIVER FIBROSIS IN CLINICALLY RELEVANT MODEL SYSTEMS. IN THIS REVIEW WE WILL DISCUSS THE ROLES OF THESE NUCLEAR CO-ACTIVATORS IN HSC ACTIVATION, THEIR MECHANISM OF ACTION IN THE FIBROTIC PROCESS IN THE LIVER AND OTHER ORGANS, AND THE POTENTIAL OF TARGETING THEIR ACTIVITY WITH SMALL MOLECULE DRUGS FOR FIBROSIS REVERSAL. 2016 3 699 31 BROMODOMAIN PROTEIN 4 IS A KEY MOLECULAR DRIVER OF TGFBETA1-INDUCED HEPATIC STELLATE CELL ACTIVATION. LIVER FIBROSIS IS CHARACTERIZED BY THE EXCESSIVE DEPOSITION OF EXTRACELLULAR MATRIX IN LIVER. CHRONIC LIVER INJURY INDUCES THE ACTIVATION OF HEPATIC STELLATE CELL (HSCS), A KEY STEP IN LIVER FIBROGENESIS. THE ACTIVATED HSC IS THE PRIMARY SOURCE OF ECM AND CONTRIBUTES SIGNIFICANTLY TO LIVER FIBROSIS. TGFBETA1 IS THE MOST POTENT PRO-FIBROTIC CYTOKINE. BROMODOMAIN PROTEIN 4 (BRD4), AN EPIGENETIC READER OF HISTONE ACETYLATION MARKS, WAS CRUCIAL FOR PROFIBROTIC GENE EXPRESSION IN HSCS. THE PRESENT STUDY AIMED TO INVESTIGATE THE ROLES OF BRD4 IN TGFBETA1-DEPENDENT HSC ACTIVATION AND LIVER FIBROSIS, FOCUSING ON TGFBETA1-INDUCED ALTERATIONS OF THE LEVELS OF THE FIBROTIC-RELATED IMPORTANT PROTEINS IN HSCS BY EMPLOYING THE HETEROZYGOUS TGFBETA1 KNOCKOUT MICE AND BRD4 KNOCKDOWN IN VIVO AND IN VITRO. RESULTS REVEALED THAT BRD4 PROTEIN LEVEL WAS SIGNIFICANTLY UPREGULATED BY TGFBETA1 AND BRD4 KNOCKDOWN REDUCED TGFBETA1-INDUCED HSC ACTIVATION AND LIVER FIBROSIS. BRD4 WAS REQUIRED FOR THE INFLUENCES OF TGFBETA1 ON PDGFBETA RECEPTOR AND ON THE PATHWAYS OF SMAD3, STAT3, AND AKT. BRD4 ALSO MEDIATED TGFBETA1-INDUCED INCREASES IN HISTONE ACETYLTRANSFERASE P300, THE PIVOTAL PRO-INFLAMMATORY NFKB P65, AND TISSUE INHIBITOR OF METALLOPROTEINASE 1 WHEREAS BRD4 REDUCED CASPASE-3 PROTEIN LEVELS IN HSCS DURING LIVER INJURY, INDEPENDENT OF TGFBETA1. FURTHER EXPERIMENTS INDICATED THE INTERACTION BETWEEN TGFBETA1-INDUCED BRD4 AND NFKB P65 IN HSCS AND IN LIVER OF TAA-INDUCED LIVER INJURY. HUMAN CIRRHOTIC LIVERS WERE DEMONSTRATED A PARALLEL INCREASE IN THE PROTEIN LEVELS OF BRD4 AND NFKB P65 IN HSCS. THIS STUDY REVEALED THAT BRD4 WAS A KEY MOLECULAR DRIVER OF TGFBETA1-INDUCED HSC ACTIVATION AND LIVER FIBROSIS. 2023 4 1572 31 DNA METHYLATION PATTERNS IN JUVENILE SYSTEMIC SCLEROSIS AND LOCALIZED SCLERODERMA. SCLERODERMA REFERS TO A GROUP OF CHRONIC FIBROTIC IMMUNE-MEDIATED DISEASES OF UNKNOWN ETIOLOGY. CHARACTERIZING EPIGENETIC CHANGES IN CHILDHOOD-ONSET SCLERODERMA, SYSTEMIC SCLEROSIS OR LOCALIZED SCLERODERMA, HAS NOT BEEN PREVIOUSLY PERFORMED. THE AIM OF THIS STUDY WAS TO ASSESS DNA METHYLATION DIFFERENCES AND SIMILARITIES BETWEEN JUVENILE SYSTEMIC SCLEROSIS (JSSC) AND JUVENILE LOCALIZED SCLERODERMA (JLS) COMPARED TO MATCHED HEALTHY CONTROLS. GENOME-WIDE DNA METHYLATION CHANGES IN PERIPHERAL BLOOD MONONUCLEAR CELL SAMPLES WERE ASSESSED USING THE METHYLATIONEPIC ARRAY FOLLOWED BY BIOINFORMATIC ANALYSIS AND LIMITED FUNCTIONAL ASSESSMENT. WE IDENTIFIED A TOTAL OF 105 AND 144 DIFFERENTIALLY METHYLATED SITES COMPARED TO HEALTHY CONTROLS IN JSSC AND JLS, RESPECTIVELY. THE MAJORITY OF DIFFERENTIALLY METHYLATED SITES AND GENES REPRESENTED WERE UNIQUE TO EITHER JSSC OR JLS SUGGESTING A DIFFERENT UNDERLYING EPIGENETIC PATTERN IN BOTH DISEASES. AMONG SHARED DIFFERENTIALLY METHYLATED GENES, METHYLATION LEVELS IN A CPG SITE IN FGFR2 CAN DISTINGUISH BETWEEN LS AND HEALTHY PBMCS WITH A HIGH ACCURACY. CANONICAL PATHWAY ANALYSIS REVEALED THAT INFLAMMATORY PATHWAYS WERE ENRICHED IN GENES DIFFERENTIALLY METHYLATED IN JSSC, INCLUDING STAT3, NF-KAPPAB, AND IL-15 PATHWAYS. IN CONTRAST, THE HIPPO SIGNALING PATHWAY WAS ENRICHED IN JLS. OUR DATA ALSO SUGGEST A POTENTIAL ROLE FOR NOTCH3 IN BOTH JSSC AND JLS, AND REVEALED A NUMBER OF TRANSCRIPTION FACTORS UNIQUE TO EACH OF THE TWO DISEASES. IN SUMMARY, OUR DATA REVEALED IMPORTANT INSIGHTS INTO JSSC AND JLS AND SUGGEST A POTENTIALLY NOVEL EPIGENETIC DIAGNOSTIC BIOMARKER FOR LS. 2021 5 1764 34 EARLY-IMMEDIATE GENE EGR1 IS ASSOCIATED WITH TGFBETA1 REGULATION OF EPIGENETIC READER BROMODOMAIN-CONTAINING PROTEIN 4 VIA THE CANONICAL SMAD3 SIGNALING IN HEPATIC STELLATE CELLS IN VITRO AND IN VIVO. UPON CHRONIC DAMAGE TO THE LIVER, MULTIPLE CYTOKINES STIMULATE HEPATIC STELLATE CELLS (HSCS), CAUSING THE ALTERATIONS OF GENE EXPRESSION PROFILES AND THUS LEADING TO HSC ACTIVATION, A KEY STEP IN LIVER FIBROGENESIS. ACTIVATED HSCS ARE THE DOMINANT CONTRIBUTORS TO LIVER FIBROSIS. BROMODOMAIN CONTAINING PROTEIN 4 (BRD4), AN IMPORTANT EPIGENETIC READER, WAS DEMONSTRATED TO CONCENTRATE ON HUNDREDS OF ENHANCERS ASSOCIATED WITH GENES INVOLVED IN MULTIPLE PROFIBROTIC PATHWAYS, THEREBY DIRECTING HSC ACTIVATION AND THE FIBROTIC RESPONSES. THE PRESENT STUDIES WERE DESIGNED TO EXAMINE THE EFFECT OF TRANSFORMING GROWTH FACTOR BETA-1 (TGFBETA1), THE MOST POTENT PRO-FIBROTIC CYTOKINE, ON BRD4 EXPRESSION IN HSCS AND, IF SO, ELUCIDATED THE UNDERLYING MECHANISMS IN VITRO AND IN VIVO. THE EXPERIMENTS EMPLOYED THE HETEROGENEOUS TGFBETA1 KNOCKOUT (TGFBETA1(+/-) ) MICE, GENE KNOCKDOWN IN VIVO, AND A MODEL OF THIOACETAMIDE (TAA)-INDUCED LIVER INJURY. THE RESULTS REVEALED THAT TGFBETA1 ENHANCED BRD4 EXPRESSION IN HSCS, WHICH WAS MEDIATED, AT LEAST, BY SMAD3 SIGNALING AND EARLY-IMMEDIATE GENE EGR1 (EARLY GROWTH RESPONSE-1). TGFBETA1-INDUCED SMAD3 SIGNALING INCREASED EGR1 EXPRESSION AND PROMOTED EGR1 BINDING TO BRD4 PROMOTER AT A SITE AROUND -111 BP, PROMOTING BRD4 EXPRESSION. EGR1 KNOCKDOWN REDUCED BRD4 EXPRESSION IN HSCS IN A MOUSE MODEL OF TAA-INDUCED LIVER INJURY AND LESSENED LIVER FIBROSIS. DOUBLE FLUORESCENCE STAINING DEMONSTRATED A STRONG INCREASE IN BRD4 EXPRESSION IN ACTIVATED HSCS IN FIBROTIC AREAS OF THE HUMAN LIVERS, PARALLELING THE UPREGULATION OF P-SMAD3 AND EGR1. THIS RESEARCH SUGGESTED NOVEL MOLECULAR EVENTS UNDERLYING THE ROLES OF THE MASTER PRO-FIBROTIC CYTOKINE TGFBETA1 IN HSC ACTIVATION AND LIVER FIBROGENESIS. 2022 6 5234 41 PROFILE ANALYSIS AND FUNCTIONAL MODELING IDENTIFY CIRCULAR RNAS IN NONALCOHOLIC FATTY LIVER DISEASE AS REGULATORS OF HEPATIC LIPID METABOLISM. NONALCOHOLIC FATTY LIVER DISEASE (NAFLD) IS THE LEADING CAUSE OF CHRONIC LIVER DISEASE, ASSOCIATED WITH AN OUTCOME OF HEPATIC FIBROSIS/CIRRHOSIS AND HEPATOCELLULAR CARCINOMA. HOWEVER, LIMITED EXPLORATION OF THE UNDERLYING MECHANISMS HINDERS ITS PREVENTION AND TREATMENT. TO INVESTIGATE THE MECHANISMS OF EPIGENETIC REGULATION IN NAFLD, THE EXPRESSION PROFILE OF CIRCULAR RNA (CIRCRNA) OF RODENTS IN WHICH NAFLD WAS INDUCED BY A HIGH-FAT, HIGH-CHOLESTEROL (HFHC) DIET WAS STUDIED. MODELING OF THE CIRCRNA-MICRORNA (MIRNA) -MRNA REGULATORY NETWORK REVEALED THE FUNCTIONAL CHARACTERISTICS OF NAFLD-SPECIFIC CIRCRNAS. THE TARGETS AND EFFECTS IN THE LIVER OF SUCH NAFLD-SPECIFIC CIRCRNAS WERE FURTHER ASSESSED. OUR RESULTS UNCOVERED THAT THE DOWNREGULATION OF 28 ANNOTATED CIRCRNAS CHARACTERIZES HFHC DIET-INDUCED NAFLD. AMONG THE DOWNREGULATED CIRCRNAS, LONG INTERGENIC NON-PROTEIN CODING RNA, P53 INDUCED TRANSCRIPT (LNCPINT) -DERIVED CIRCRNAS (CIRC_0001452, CIRC_0001453, AND CIRC_0001454) TARGETED BOTH MIR-466I-3P AND MIR-669C-3P. THEIR DEFICIENCY IN NAFLD ABROGATED THE CIRCRNA-BASED INHIBITORY EFFECT ON BOTH MIRNAS, WHICH FURTHER INACTIVATED THE AMPK SIGNALING PATHWAY VIA AMPK-ALPHA1 SUPPRESSION. INHIBITION OF THE AMPK SIGNALING PATHWAY PROMOTES HEPATIC STEATOSIS, DEPENDING ON THE TRANSCRIPTIONAL AND TRANSLATIONAL UPREGULATION OF LIPOGENIC GENES, SUCH AS THOSE ENCODING STEROL REGULATORY ELEMENT-BINDING PROTEIN 1 (SREBP1) AND FATTY ACID SYNTHASE (FASN) IN HEPATOCYTES. THE LEVELS OF LNCPINT-DERIVED CIRCRNAS DISPLAYED A NEGATIVE ASSOCIATION WITH HEPATIC TRIGLYCERIDE (TG) CONCENTRATION. THESE FINDINGS SUGGEST THAT LOSS OF LNCPINT-DERIVED CIRCRNAS MAY UNDERLIE NAFLD VIA MIR-466I-3P- AND MIR-669C-3P-DEPENDENT INACTIVATION OF THE AMPK SIGNALING PATHWAY. 2022 7 4097 20 MATRIX STIFFNESS REGULATES MACROPHAGE POLARIZATION IN ATHEROSCLEROSIS. ATHEROSCLEROSIS IS A CHRONIC INFLAMMATORY DISEASE AND THE PATHOLOGICAL BASIS OF MANY FATAL CARDIOVASCULAR DISEASES. MACROPHAGES, THE MAIN INFLAMMATORY CELLS IN ATHEROSCLEROTIC PLAQUE, HAVE A PARADOX ROLE IN DISEASE PROGRESSION. IN RESPONSE TO DIFFERENT MICROENVIRONMENTS, MACROPHAGES MAINLY HAVE TWO POLARIZED DIRECTIONS: PRO-INFLAMMATORY MACROPHAGES AND ANTI-INFLAMMATORY MACROPHAGES. MORE AND MORE EVIDENCE SHOWS THAT MACROPHAGE IS MECHANOSENSITIVE AND MATRIX STIFFNESS REGULATE MACROPHAGE PHENOTYPES IN ATHEROSCLEROSIS. HOWEVER, THE MOLECULAR MECHANISM OF MATRIX STIFFNESS REGULATING MACROPHAGE POLARIZATION STILL LACKS IN-DEPTH RESEARCH, WHICH HINDERS THE DEVELOPMENT OF NEW ANTI-ATHEROSCLEROTIC THERAPIES. IN THIS REVIEW, WE DISCUSS THE IMPORTANT ROLE OF MATRIX STIFFNESS IN REGULATING MACROPHAGE POLARIZATION THROUGH MECHANICAL SIGNAL TRANSDUCTION (HIPPO, PIEZO, CYTOSKELETON, AND INTEGRIN) AND EPIGENETIC MECHANISMS (MIRNA, DNA METHYLATION, AND HISTONE). WE HOPE TO PROVIDE A NEW PERSPECTIVE FOR ATHEROSCLEROSIS THERAPY BY TARGETING MATRIX STIFFNESS AND MACROPHAGE POLARIZATION. 2022 8 5120 35 POSSIBLE EPIGENETIC REGULATORY EFFECT OF DYSREGULATED CIRCULAR RNAS IN EPILEPSY. CIRCULAR RNAS (CIRCRNAS) INVOLVE IN THE EPIGENETIC REGULATION AND ITS MAJOR MECHANISM IS THE SEQUESTRATION OF THE TARGET MICRO RNAS (MIRNAS). WE HYPOTHESIZED THAT CIRCRNAS MIGHT BE RELATED WITH THE PATHOPHYSIOLOGY OF CHRONIC EPILEPSY AND EVALUATED THE ALTERED CIRCRNA EXPRESSIONS AND THEIR POSSIBLE REGULATORY EFFECTS ON THEIR TARGET MIRNAS AND MRNAS IN A MOUSE EPILEPSY MODEL. THE CIRCRNA EXPRESSION PROFILE IN THE HIPPOCAMPUS OF THE PILOCARPINE MICE WAS ANALYZED AND COMPARED WITH CONTROL. THE CORRELATION BETWEEN THE EXPRESSION OF MIRNA BINDING SITES (MIRNA RESPONSE ELEMENTS, MRE) IN THE DYSREGULATED CIRCRNAS AND THE EXPRESSION OF THEIR TARGET MIRNAS WAS EVALUATED. AS MIRNAS ALSO INHIBIT THEIR TARGET MRNAS, CIRCRNA-MIRNA-MRNA REGULATORY NETWORK, COMPRISED OF DYSREGULATED RNAS THAT TARGETS ONE ANOTHER WERE SEARCHED. FOR THE IDENTIFIED NETWORKS, BIOINFORMATICS ANALYSES WERE PERFORMED. AS THE RESULT, FORTY-THREE CIRCRNAS WERE DYSREGULATED IN THE HIPPOCAMPUS (UP-REGULATED, 26; DOWN-REGULATED, 17). THE CHANGE IN THE EXPRESSION OF MRE IN THOSE CIRCRNAS NEGATIVELY CORRELATED WITH THE CHANGE IN THE RELEVANT TARGET MIRNA EXPRESSION (R = -0.461, P<0.001), SUPPORTING THAT CIRCRNAS INHIBIT THEIR TARGET MIRNA. 333 DYSREGULATED CIRCRNA-MIRNA-MRNA NETWORKS WERE IDENTIFIED. GENE ONTOLOGY AND PATHWAY ANALYSES DEMONSTRATED THAT THE UP-REGULATED MRNAS IN THOSE NETWORKS WERE CLOSELY RELATED TO THE MAJOR PROCESSES IN EPILEPSY. AMONG THEM, STRING ANALYSIS IDENTIFIED 37 KEY MRNAS WITH ABUNDANT (>/=4) INTERACTIONS WITH OTHER DYSREGULATED TARGET MRNAS. THE DYSREGULATION OF THE CIRCRNAS WHICH HAD MULTIPLE INTERACTIONS WITH KEY MRNAS WERE VALIDATED BY PCR. WE CONCLUDED THAT DYSREGULATED CIRCRNAS MIGHT HAVE A PATHOPHYSIOLOGIC ROLE IN CHRONIC EPILEPSY BY REGULATING MULTIPLE DISEASE RELEVANT MRNAS VIA CIRCRNA-MIRNA-MRNA INTERACTIONS. 2018 9 4286 46 MICRORNA EXPRESSION ANALYSIS IN HIGH FAT DIET-INDUCED NAFLD-NASH-HCC PROGRESSION: STUDY ON C57BL/6J MICE. BACKGROUND: HEPATOCELLULAR CARCINOMA (HCC) IS THE MOST COMMON MALIGNANT TUMOR OF THE LIVER. NON-ALCOHOLIC FATTY LIVER DISEASE (NAFLD) IS A FREQUENT CHRONIC LIVER DISORDER IN DEVELOPED COUNTRIES. NAFLD CAN PROGRESS THROUGH THE MORE SEVERE NON ALCOHOLIC STEATOHEPATITIS (NASH), CIRRHOSIS AND, LASTLY, HCC. GENETIC AND EPIGENETIC ALTERATIONS OF CODING GENES AS WELL AS DEREGULATION OF MICRORNAS (MIRNAS) ACTIVITY PLAY A ROLE IN HCC DEVELOPMENT. IN THIS STUDY, THE C57BL/6J MOUSE MODEL WAS LONG TERM HIGH-FAT (HF) OR LOW-FAT (LF) DIET FED, IN ORDER TO ANALYZE MOLECULAR MECHANISMS RESPONSIBLE FOR THE HEPATIC DAMAGE PROGRESSION. METHODS: MICE WERE HF OR LF DIET FED FOR DIFFERENT TIME POINTS, THEN PLASMA AND HEPATIC TISSUES WERE COLLECTED. HISTOLOGICAL AND CLINICAL CHEMISTRY ASSAYS WERE PERFORMED TO ASSESS THE PROGRESSION OF LIVER DISEASE. MICRORNAS' DIFFERENTIAL EXPRESSION WAS EVALUATED ON POOLED RNAS FROM TISSUES, AND SOME MIRNAS SHOWING DYSREGULATION WERE FURTHER ANALYZED AT THE INDIVIDUAL LEVEL. RESULTS: CHOLESTEROL, LOW AND HIGH DENSITY LIPOPROTEINS, TRIGLYCERIDES AND ALANINE AMINOTRANSFERASE INCREASE WAS DETECTED IN HF MICE. GROSS ANATOMICAL EXAMINATION REVEALED HEPATOMEGALY IN HF LIVERS, AND HISTOLOGICAL ANALYSIS HIGHLIGHTED DIFFERENT DEGREES AND LEVELS OF STEATOSIS, INFLAMMATORY INFILTRATE AND FIBROSIS IN HF AND LF ANIMALS, DEMONSTRATING THE PROGRESSION FROM NAFLD THROUGH NASH. MACROSCOPIC NODULES, SHOWING TYPICAL NEOPLASTIC FEATURES, WERE OBSERVED IN 20% OF HF DIET FED MICE. FIFTEEN MIRNAS DIFFERENTIALLY EXPRESSED IN HF WITH RESPECT TO LF HEPATIC TISSUES DURING THE PROGRESSION OF LIVER DAMAGE, AND IN TUMORS WITH RESPECT TO HF NON TUMOR LIVER SPECIMENS WERE IDENTIFIED. AMONG THEM, MIR-340-5P, MIR-484, MIR-574-3P, MIR-720, WHOSE EXPRESSION WAS NEVER DESCRIBED IN NAFLD, NASH AND HCC TISSUES, AND MIR-125A-5P AND MIR-182, WHICH SHOWED EARLY AND SIGNIFICANT DYSREGULATION IN THE SEQUENTIAL HEPATIC DAMAGE PROCESS. CONCLUSIONS: IN THIS STUDY, FIFTEEN MICRORNAS WHICH WERE MODULATED IN HEPATIC TISSUES AND IN TUMORS DURING THE TRANSITION NAFLD-NASH-HCC ARE REPORTED. BESIDES SOME ALREADY DESCRIBED, NEW AND EARLY DYSREGULATED MIRNAS WERE IDENTIFIED. FUNCTIONAL ANALYSES ARE NEEDED TO VALIDATE THE RESULTS HERE OBTAINED, AND TO BETTER DEFINE THE ROLE OF THESE MOLECULES IN THE PROGRESSION OF THE HEPATIC DISEASE. 2016 10 3752 42 INTEGRATED ANALYSIS OF CIRCRNAS AND MRNAS EXPRESSION PROFILE REVEALED THE INVOLVEMENT OF HSA_CIRC_0007919 IN THE PATHOGENESIS OF ULCERATIVE COLITIS. BACKGROUND: ULCERATIVE COLITIS (UC) IS CHARACTERIZED BY CHRONIC INFLAMMATION IN THE COLON AND EPIGENETIC FACTORS UNDERLYING THE OCCURRENCE. CIRCULAR RNAS (CIRCRNAS) HAVE BEEN UNDER INTENSIVE FOCUS DUE TO THE CIRCULAR CONSTRUCT AND GENE-REGULATING FUNCTIONS. HOWEVER, THE CHANGES AND ROLES OF CIRCRNAS IN UC REMAIN UNKNOWN. METHODS: MICROARRAYS WERE USED TO DETECT THE DIFFERENTIALLY EXPRESSED GENES, AND QUANTITATIVE REAL-TIME PCR WAS USED TO IDENTIFY THE CHANGES IN UC. IN SILICO ANALYSES WERE PERFORMED TO PREDICT THE FUNCTIONS OF CIRCRNAS AND MRNAS. IN VITRO, EPITHELIAL CELL LINES WERE STIMULATED BY PRO-INFLAMMATION EFFECTORS TO TEST THE ALTERATIONS IN CIRCRNAS. CIRCRNAS-MICRORNAS-MRNAS NETWORK CLARIFIED THE POTENTIAL MECHANISMS UNDERLYING CIRCRNAS IN UC. THE BINDING SITE BETWEEN HSA_CIRC_0007919 AND MIR-138 OR LET-7A WAS VERIFIED USING DUAL-LUCIFERASE ASSAY. RESULTS: A TOTAL OF 264 SIGNIFICANTLY DYSREGULATED CIRCRNAS AND 1869 DIFFERENTIALLY EXPRESSED MRNAS IN INFLAMED MUCOSA WERE COMPARED WITH THE NON-INFLAMED MUCOSA IN UC. HSA_CIRC_0004662 AND HSA_CIRC_0007919 WERE ALTERED LARGELY IN UC TISSUES. HSA_CIRC_0007919 WAS REDUCED PERSISTENTLY AFTER INFLAMMATORY TREATMENTS, AND IT WAS RELEVANT TO MAYO ENDOSCOPIC SUBSCORES AND THE EXPRESSION OF TIGHT JUNCTION MOLECULES. FINALLY, HSA_CIRC_0007919 COULD HARBOR MIR-138, AND LET-7A TO REGULATE THE TARGETED MRNAS EPC1 AND VIPR1. CONCLUSIONS: SEVERAL CIRCRNAS WERE DIFFERENTIALLY EXPRESSED IN UC. HSA_CIRC_0007919 IS RELATED TO CLINICAL CHARACTERISTICS AND EPITHELIAL INTEGRITY BY BINDING TO HSA-LET-7A, HSA-MIR-138 TO REGULATE THE TARGET GENES. CIRCRNAS, ESPECIALLY HSA_CIRC_0007919, ARE ASSOCIATED WITH THE PATHOGENESIS AND DEVELOPMENT OF UC, WITH POTENTIAL DIAGNOSTIC AND THERAPEUTIC IMPLICATIONS. 2019 11 4303 39 MICRORNA-223 INHIBITS TISSUE FACTOR EXPRESSION IN VASCULAR ENDOTHELIAL CELLS. OBJECTIVE: ATHEROSCLEROSIS IS A CHRONIC INFLAMMATORY PROCESS, IN WHICH VASCULAR ENDOTHELIAL CELLS (ECS) BECOME DYSFUNCTIONAL OWING TO THE EFFECTS OF CHEMICAL SUBSTANCES, SUCH AS INFLAMMATORY FACTOR AND GROWTH FACTORS. TISSUE FACTOR (TF) EXPRESSION IS INDUCED BY THE ABOVE CHEMICAL SUBSTANCES IN ACTIVATED ECS. TF INITIATES THROMBOSIS ON DISRUPTED ATHEROSCLEROTIC PLAQUES WHICH PLAYS AN ESSENTIAL ROLE DURING THE ONSET OF ACUTE CORONARY SYNDROMES (ACS). INCREASING EVIDENCES SUGGEST THE IMPORTANT ROLE OF MICRORNAS AS EPIGENETIC REGULATORS OF ATHEROSCLEROTIC DISEASE. THE AIM OF OUR STUDY IS TO IDENTIFY IF MICRORNA-223 (MIR-223) TARGETS TF IN ECS. METHODS AND RESULTS: BIOINFORMATIC ANALYSIS SHOWED THAT TF IS A TARGET CANDIDATE OF MIR-223. WESTERN BLOTTING ANALYSIS REVEALED THAT TUMOR NECROSIS FACTOR ALPHA (TNF-ALPHA) INCREASED TF EXPRESSION IN AORTA OF C57BL/6J MICE AND CULTURED ECS (EA.HY926 CELLS AND HUVEC) AFTER 4 H TREATMENT. IN TNF-ALPHA TREATED ECS, TF MRNA WAS ALSO INCREASED MEASURED BY REAL-TIME PCR. REAL-TIME PCR RESULTS SHOWED THAT MIR-223 LEVELS WERE DOWNREGULATED IN TNF-ALPHA-TREATED AORTA OF C57BL/6J MICE AND CULTURED ECS. TRANSFECTION OF ECS WITH MIR-223 MIMIC OR MIR-223 INHIBITOR MODIFIED TF EXPRESSION BOTH IN MRNA AND PROTEIN LEVELS. LUCIFERASE ASSAYS CONFIRMED THAT MIR-223 SUPPRESSED TF EXPRESSION BY BINDING TO THE SEQUENCE OF TF 3'-UNTRANSLATED REGIONS (3'UTR). TF PROCOAGULANT ACTIVITY WAS INHIBITED BY OVEREXPRESSING MIR-223 WITH OR WITHOUT TNF-ALPHA STIMULATION. CONCLUSIONS: MIR-223-MEDIATED SUPPRESSION OF TF EXPRESSION PROVIDES A NOVEL MOLECULAR MECHANISM FOR THE REGULATION OF COAGULATION CASCADE, AND SUGGESTS A CLUE AGAINST THROMBOGENESIS DURING THE PROCESS OF ATHEROSCLEROTIC PLAQUE RUPTURE. 2014 12 1666 44 DOWNREGULATION OF MICRORNA-145A-5P PROMOTES STEATOSIS-TO-NASH PROGRESSION THROUGH UPREGULATION OF NR4A2. BACKGROUND & AIMS: THE MOLECULAR MECHANISMS UNDERLYING THE PROGRESSION OF SIMPLE STEATOSIS TO NON-ALCOHOLIC STEATOHEPATITIS (NASH) REMAIN INCOMPLETELY UNDERSTOOD, THOUGH THE POTENTIAL ROLE OF EPIGENETIC REGULATION BY MICRORNA (MIRNAS) IS AN AREA OF INCREASING INTEREST. IN THE PRESENT STUDY, WE AIMED TO INVESTIGATE THE ROLE AND MECHANISM OF MIRNAS DURING STEATOSIS-TO-NASH PROGRESSION. METHODS: MIR-145A-5P WAS IDENTIFIED AS AN IMPORTANT CHECKPOINT IN STEATOSIS-TO-NASH PROGRESSION. IN VIVO LOSS-OF-FUNCTION AND GAIN-OF-FUNCTION STUDIES WERE PERFORMED TO EXPLORE THE ROLE OF MIR-145A-5P AND NR4A2 IN NASH PROGRESSION. RNA-SEQUENCING AND BIOINFORMATIC ANALYSIS WERE USED TO INVESTIGATE THE TARGETS OF MIR-145A-5P. RESULTS: SUPPRESSION OF MIR-145A-5P IN THE LIVER AGGRAVATED LIPID ACCUMULATION AND ACTIVATED HEPATIC INFLAMMATION, LIVER INJURY AND FIBROSIS IN STEATOTIC MICE, WHEREAS ITS RESTORATION MARKEDLY ATTENUATED DIET-INDUCED NASH PATHOGENESIS. MECHANISTICALLY, MIR-145A-5P WAS ABLE TO DOWNREGULATE THE NUCLEAR RECEPTOR NR4A2 AND THUS INHIBIT THE EXPRESSION OF NASH-ASSOCIATED GENES. SIMILARLY, NR4A2 OVEREXPRESSION PROMOTED STEATOSIS-TO-NASH PROGRESSION WHILE LIVER-SPECIFIC NR4A2 KNOCKOUT MICE WERE PROTECTED FROM DIET-INDUCED NASH. THIS ROLE OF THE MIR-145A-5P/NR4A2 REGULATORY AXIS WAS ALSO CONFIRMED IN PRIMARY HUMAN HEPATOCYTES. FURTHERMORE, THE EXPRESSION OF MIR-145A-5P WAS REDUCED AND THE EXPRESSION OF NR4A2 WAS INCREASED IN THE LIVERS OF PATIENTS WITH NASH, WHILE THEIR EXPRESSION LEVELS SIGNIFICANTLY NEGATIVELY AND POSITIVELY CORRELATED WITH FEATURES OF LIVER PATHOLOGY, RESPECTIVELY. CONCLUSIONS: OUR FINDINGS HIGHLIGHT THE ROLE OF THE MIR-145A-5P/NR4A2 REGULATORY AXIS IN STEATOSIS-TO-NASH PROGRESSION, SUGGESTING THAT EITHER SUPPLEMENTATION OF MIR-145A-5P OR PHARMACOLOGICAL INHIBITION OF NR4A2 IN HEPATOCYTES MAY PROVIDE A PROMISING THERAPEUTIC APPROACH FOR THE TREATMENT OF NASH. IMPACT AND IMPLICATIONS: NON-ALCOHOLIC FATTY LIVER DISEASE (NAFLD) IS A DYNAMIC SPECTRUM OF CHRONIC LIVER DISEASES RANGING FROM SIMPLE STEATOSIS TO NON-ALCOHOLIC STEATOHEPATITIS (NASH). UNFORTUNATELY, THERE ARE CURRENTLY NO APPROVED DRUGS FOR NASH. OUR CURRENT STUDY IDENTIFIED MIR-145A-5P AS A NOVEL REGULATOR THAT INHIBITS STEATOSIS-TO-NASH PROGRESSION. WE FOUND THAT MIR-145A-5P WAS ABLE TO DOWNREGULATE THE NUCLEAR RECEPTOR NR4A2 TO SUPPRESS THE EXPRESSION OF NASH-ASSOCIATED GENES. THE DIFFERENTIAL EXPRESSION OF MIR-145A-5P AND NR4A2 WAS FURTHER CONFIRMED IN PATIENTS WITH NASH, RAISING THE POSSIBILITY THAT SUPPLEMENTATION OF MIR-145A-5P OR SUPPRESSION OF NR4A2 IN HEPATOCYTES MIGHT PROVIDE NOVEL STRATEGIES FOR TREATING NASH. 2023 13 692 35 BRD4 PROMOTES HEPATIC STELLATE CELLS ACTIVATION AND HEPATIC FIBROSIS VIA MEDIATING P300/H3K27AC/PLK1 AXIS. HEPATIC FIBROSIS (HF) IS A REVERSIBLE WOUND-HEALING RESPONSE CHARACTERIZED BY EXCESSIVE EXTRACELLULAR MATRIX (ECM) DEPOSITION AND SECONDARY TO PERSISTENT CHRONIC INJURY. BROMODOMAIN PROTEIN 4 (BRD4) COMMONLY FUNCTIONS AS A "READER" TO REGULATE EPIGENETIC MODIFICATIONS INVOLVED IN VARIOUS BIOLOGICAL AND PATHOLOGICAL EVENTS, BUT THE MECHANISM OF HF REMAINS UNCLEAR. IN THIS STUDY, WE ESTABLISHED A CCL(4)-INDUCED HF MODEL AND SPONTANEOUS RECOVERY MODEL IN MICE AND FOUND ABERRANT BRD4 EXPRESSION, WHICH WAS CONSISTENT WITH THE RESULTS IN HUMAN HEPATIC STELLATE CELLS (HSCS)- LX2 CELLS IN VITRO. SUBSEQUENTLY, WE FOUND THAT DISTRICTION AND INHIBITION OF BRD4 RESTRAINED TGFBETA-INDUCED TRANS-DIFFERENTIATION OF LX2 CELLS INTO ACTIVATED, PROLIFERATIVE MYOFIBROBLASTS AND ACCELERATED APOPTOSIS, AND BRD4 OVEREXPRESSION BLOCKED MDI-INDUCED LX2 CELLS INACTIVATION AND PROMOTED THE PROLIFERATION AND INHIBITED APOPTOSIS OF INACTIVATED CELLS. ADDITIONALLY, ADENO-ASSOCIATED VIRUS SEROTYPE 8-LOADED SHORT HAIRPIN RNA-MEDIATED BRD4 KNOCKDOWN IN MICE SIGNIFICANTLY ATTENUATED CCL(4)-INDUCED FIBROTIC RESPONSES INCLUDING HSCS ACTIVATION AND COLLAGEN DEPOSITION. MECHANISTICALLY, BRD4 DEFICIENCY INHIBITED PLK1 EXPRESSION IN ACTIVATED LX2 CELLS, AND CHIP AND CO-IP ASSAYS REVEALED THAT BRD4 REGULATION OF PLK1 WAS DEPENDENT ON P300-MEDIATED ACETYLATION MODIFICATION FOR H3K27 ON THE PLK1 PROMOTER. IN CONCLUSION, BRD4 DEFICIENCY IN THE LIVER ALLEVIATES CCL(4)-INDUCED HF IN MICE, AND BRD4 PARTICIPATES IN THE ACTIVATION AND REVERSAL OF HSCS THROUGH POSITIVELY REGULATING THE P300/H3K27AC/PLK1 AXIS, PROVIDING A POTENTIAL INSIGHT FOR HF THERAPY. 2023 14 4501 24 MORPHOGENS AND HEPATIC STELLATE CELL FATE REGULATION IN CHRONIC LIVER DISEASE. HEPATIC STELLATE CELLS (HSC) ARE THE LIVER MESENCHYMAL CELL TYPE WHICH RESPONDS TO HEPATOCELLULAR DAMAGE AND PARTICIPATES IN WOUND HEALING. ALTHOUGH HSC MYOFIBROBLASTIC TRANS-DIFFERENTIATION (ACTIVATION) IS IMPLICATED IN EXCESSIVE EXTRACELLULAR MATRIX DEPOSITION, MOLECULAR UNDERSTANDING OF THIS PHENOTYPIC SWITCH FROM THE VIEWPOINT OF CELL FATE REGULATION IS LIMITED. RECENT STUDIES DEMONSTRATE THE ROLES OF ANTI-ADIPOGENIC MORPHOGENS (WNT, NECDIN, SHH) IN EPIGENETIC REPRESSION OF THE HSC DIFFERENTIATION GENE PPARGAMMA AS A CAUSAL EVENT IN HSC ACTIVATION. THESE MORPHOGENS HAVE POSITIVE CROSS-INTERACTIONS WHICH CONVERGE TO EPIGENETIC REPRESSION OF PPARGAMMA INVOLVING THE METHYL-CPG BINDING PROTEIN MECP2. HOWEVER, THESE MORPHOGENS EXPRESSED BY ACTIVATED HSC MAY ALSO PARTICIPATE IN CROSS-TALK BETWEEN HSC AND HEPATOBLASTS/HEPATOCYTES TO SUPPORT LIVER REGENERATION, AND THEIR ABERRANT REGULATION MAY CONTRIBUTE TO LIVER TUMORIGENESIS. IMPLICATIONS OF HSC-DERIVED MORPHOGENS IN THESE POSSIBILITIES ARE DISCUSSED. 2012 15 4400 39 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 16 3931 16 LIVER INJURY AND THE ACTIVATION OF THE HEPATIC MYOFIBROBLASTS. LIVER FIBROSIS IS A WOUND HEALING PROCESS, THE END RESULT OF CHRONIC LIVER INJURY ELICITED BY DIFFERENT NOXIOUS STIMULI. ACTIVATED HEPATIC STELLATE CELLS OR MYOFIBROBLASTS AND PORTAL MYOFIBROBLASTS ARE CONSIDERED AS THE MAIN PRODUCERS OF THE EXTRACELLULAR MATRIX IN THE LIVER. UPON LIVER INJURY THE QUIESCENT STELLATE CELLS TRANSDIFFERENTIATE INTO MYOFIBROBLASTS A PROCESS HIGHLIGHTED BY THE LOSS OF VITAMIN A STORES, UPREGULATION OF INTERSTITIAL TYPE COLLAGENS, SMOOTH MUSCLE ALPHA ACTIN, MATRIX METALLOPROTEINASES, PROTEOGLYCANS, AND THE INDUCTION OF CELL SURVIVAL PATHWAYS. ACTIVATION OF HEPATIC STELLATE CELLS IS A RESULT OF A COMPLEX INTERPLAY BETWEEN THE PARENCHYMAL CELLS, IMMUNE CELLS, EXTRACELLULAR MATRIX MECHANICS AND EXTRAHEPATIC MILIEU SUCH AS THE GUT MICROBIOME. IN THIS REVIEW WE WILL FOCUS ON THE PATHOMECHANISM OF STELLATE CELL ACTIVATION FOLLOWING CHRONIC LIVER INJURY; WITH THE AIM OF IDENTIFYING POSSIBLE TREATMENT TARGETS FOR ANTI-FIBROGENIC AGENTS. 2013 17 3944 41 LNCRNA H19-EZH2 INTERACTION PROMOTES LIVER FIBROSIS VIA REPROGRAMMING H3K27ME3 PROFILES. LIVER FIBROSIS IS A WOUND-HEALING PROCESS CHARACTERIZED BY EXCESS FORMATION OF EXTRACELLULAR MATRIX (ECM) FROM ACTIVATED HEPATIC STELLATE CELLS (HSCS). PREVIOUS STUDIES SHOW THAT BOTH EZH2, AN EPIGENETIC REGULATOR THAT CATALYZES LYSINE 27 TRIMETHYLATION ON HISTONE 3 (H3K27ME3), AND LONG NON-CODING RNA H19 ARE HIGHLY CORRELATED WITH FIBROGENESIS. IN THE CURRENT STUDY, WE INVESTIGATED THE UNDERLYING MECHANISMS. VARIOUS MODELS OF LIVER FIBROSIS INCLUDING MDR2(-/-), BILE DUCT LIGATION (BDL) AND CCL(4) MICE WERE ADAPTED. WE FOUND THAT EZH2 WAS MARKEDLY UPREGULATED AND CORRELATED WITH H19 AND FIBROTIC MARKERS EXPRESSION IN THESE MODELS. ADMINISTRATION OF EZH2 INHIBITOR 3-DZNEP CAUSED SIGNIFICANT PROTECTIVE EFFECTS IN THESE MODELS. FURTHERMORE, TREATMENT WITH 3-DZNEP OR GSK126 SIGNIFICANTLY INHIBITED PRIMARY HSC ACTIVATION AND PROLIFERATION IN TGF-BETA-TREATED HSCS AND H19-OVEREXPREESING LX2 CELLS IN VIVO. USING RNA-PULL DOWN ASSAY COMBINED WITH RNA IMMUNOPRECIPITATION, WE DEMONSTRATED THAT H19 COULD DIRECTLY BIND TO EZH2. INTEGRATED ANALYSIS OF RNA-SEQUENCING (RNA-SEQ) AND CHROMATIN IMMUNOPRECIPITATION SEQUENCING (CHIP-SEQ) FURTHER REVEALED THAT H19 REGULATED THE REPROGRAMMING OF EZH2-MEDIATED H3K27ME3 PROFILES, WHICH EPIGENETICALLY PROMOTED SEVERAL PATHWAYS FAVORING HSCS ACTIVATION AND PROLIFERATION, INCLUDING EPITHELIAL-MESENCHYMAL TRANSITION AND WNT/BETA-CATENIN SIGNALING. IN CONCLUSION, HIGHLY EXPRESSED H19 IN CHRONIC LIVER DISEASES PROMOTES FIBROGENESIS BY REPROGRAMMING EZH2-MEDIATED EPIGENETIC REGULATION OF HSCS ACTIVATION. TARGETING THE H19-EZH2 INTERACTION MAY SERVE AS A NOVEL THERAPEUTIC APPROACH FOR LIVER FIBROSIS. 2023 18 3946 32 LNCRNA MALAT1 BINDS CHROMATIN REMODELING SUBUNIT BRG1 TO EPIGENETICALLY PROMOTE INFLAMMATION-RELATED HEPATOCELLULAR CARCINOMA PROGRESSION. HEPATOCELLULAR CARCINOMA (HCC) IS ONE TYPE OF CANCERS WHOSE CARCINOGENESIS AND PROGRESSION ARE CLOSELY RELATED TO CHRONIC INFLAMMATION. IDENTIFYING THE MOLECULAR MECHANISMS FOR INFLAMMATION-RELATED HCC PROGRESSION WILL CONTRIBUTE TO IMPROVE THE EFFICACY OF CURRENT THERAPEUTICS FOR HCC PATIENTS. MANY KINDS OF EPIGENETIC FACTORS, INCLUDING LONG NON-CODING RNAS (LNCRNAS), HAVE BEEN DISCOVERED TO BE IMPORTANT IN HCC GROWTH AND METASTASIS. HOWEVER, HOW THE LNCRNAS PROMOTE HCC PROGRESSION AND WHAT'S THE APPLICATION OF LNCRNA SILENCING IN VIVO IN SUPPRESSING HCC REMAIN TO BE FURTHER INVESTIGATED. HERE, WE FOUND THAT LNCRNA METASTASIS ASSOCIATED LUNG ADENOCARCINOMA TRANSCRIPT1 (MALAT1) WAS UPREGULATED IN HCC TUMOR TISSUES, AND KNOCKDOWN OF MALAT1 SUPPRESSED PROLIFERATION, CELL CYCLE AND INVASION OF HCC CELLS IN RESPONSE TO LIPOPOLYSACCHARIDE (LPS) STIMULATION. KNOCKDOWN OF MALAT1 SIGNIFICANTLY INHIBITED LPS-INDUCED PRO-INFLAMMATORY MEDIATORS IL-6 AND CXCL8 EXPRESSION IN HCC CELLS, WHICH COULD BE RESTORED BY OVEREXPRESSING MALAT1. MECHANISTICALLY, MALAT1 RECRUITED BRAHMA-RELATED GENE 1 (BRG1), A CATALYTIC SUBUNIT OF CHROMATIN REMODELING COMPLEX SWITCHING/SUCROSE NON-FERMENTABLE (SWI/SNF), TO THE PROMOTER REGION OF IL-6 AND CXCL8, AND THUS FACILITATED NF-KAPPAB TO INDUCE THE EXPRESSION OF THESE INFLAMMATORY FACTORS. IMPORTANTLY, IN VIVO SILENCING OF MALAT1 IN HCC TISSUES INHIBITED GROWTH OF HCC XENOGRAFTS, AND ALSO SUPPRESSED THE EXPRESSION OF PRO-INFLAMMATORY FACTORS IN HCC TISSUES ACCORDINGLY. OUR RESULTS DEMONSTRATE THAT MALAT1 PROMOTES HCC PROGRESSION BY BINDING BRG1 TO EPIGENETICALLY ENHANCE INFLAMMATORY RESPONSE IN HCC TISSUES, AND SILENCING OF MALAT1 MAY BE A POTENTIAL APPROACH TO THE TREATMENT OF HCC. 2019 19 1021 24 CIRCULAR RNA AS AN EPIGENETIC REGULATOR IN CHRONIC LIVER DISEASES. CIRCULAR RNA (CIRCRNA) IS A TYPE OF NON-CODING RNA CHARACTERIZED BY A COVALENTLY CLOSED CONTINUOUS LOOP. CIRCRNA IS GENERATED BY PRE-MRNA THROUGH BACK-SPLICING AND IS PROBABLY CLEARED UP BY EXTRACELLULAR VESICLES. CIRCRNAS PLAY A PIVOTAL ROLE IN THE EPIGENETIC REGULATION OF GENE EXPRESSION AT TRANSCRIPTIONAL AND POST-TRANSCRIPTIONAL LEVELS. RECENTLY, CIRCRNAS HAVE BEEN DEMONSTRATED TO BE INVOLVED IN THE REGULATION OF LIVER HOMEOSTASIS AND DISEASES. HOWEVER, THE EPIGENETIC ROLE AND UNDERLYING MECHANISMS OF CIRCRNAS IN CHRONIC LIVER DISEASES REMAIN UNCLEAR. THIS REVIEW DISCUSSED THE ROLE OF CIRCRNAS IN NON-NEOPLASTIC CHRONIC LIVER DISEASES, INCLUDING ALCOHOLIC LIVER DISEASE (ALD), METABOLIC-ASSOCIATED FATTY LIVER DISEASE (MAFLD), VIRAL HEPATITIS, LIVER INJURY AND REGENERATION, LIVER CIRRHOSIS, AND AUTOIMMUNE LIVER DISEASE. THE REVIEW ALSO HIGHLIGHTED THAT FURTHER EFFORTS ARE URGENTLY NEEDED TO DEVELOP CIRCRNAS AS NOVEL DIAGNOSTICS AND THERAPEUTICS FOR CHRONIC LIVER DISEASES. 2021 20 3947 41 LNCRNA UCA1 INDUCES AUTOPHAGIC GENE EXPRESSION VIA EPIGENETIC REGULATION MEDIATED BY ATG16L1 AND MIR-132-3P IN SH-SY5Y CELLS TREATED WITH RETINOIC ACID. OBJECTIVE: EPILEPSY IS A CHRONIC BRAIN DISEASE WITH RECURRENT SEIZURES. AUTOPHAGY PLAYS A CRUCIAL ROLE IN THE PROGRESSION OF EPILEPSY. THIS STUDY AIMED TO EXPLORE THE FUNCTION AND INTRINSIC MECHANISM OF THE LONG NON-CODING RNA (LNCRNA) UCA1/MIR-132-3P/ATG16L1 AXIS IN EPILEPSY VIA REGULATION OF AUTOPHAGY. METHODS: THE EXPRESSION OF LNCRNA UCA1, MIR-132-3P AND ATG16L1 WAS MEASURED IN SERUM FROM EPILEPTIC PATIENTS BY QUANTITATIVE RT-PCR. A SH-SY5Y CELL MODEL WAS FURTHER CONSTRUCTED USING RETINOIC ACID TO INVESTIGATE THE UCA1/ MIR-132-3P/ATG16L1 AXIS BY QUANTITATIVE RT-PCR, WESTERN BLOTTING, FLUORESCENCE IN SITU HYBRIDISATION, RNA IMMUNOPRECIPITATION, CHROMATIN IMMUNOPRECIPITATION, AND A DUAL-LUCIFERASE REPORTER GENE ASSAY. RESULTS: IN THE SERUM OF EPILEPTIC PATIENTS, THE LEVEL OF LNCRNA UCA1 AND ATG16L1 WAS REDUCED AND MIR-132-3P ELEVATED, COMPARED TO CONTROLS. SIMILARLY, IN THE SH-SY5Y CELL MODEL, THE LEVEL OF LNCRNA UCA1 AND ATG16L1 WAS REDUCED AND MIR-132-3P ELEVATED IN RETINOIC ACID-TREATED CELLS; LNCRNA UCA1 WAS MAINLY LOCATED IN THE CYTOPLASM. LNCRNA UCA1 OVEREXPRESSION WAS SHOWN TO PROMOTE AUTOPHAGIC GENE EXPRESSION, WHICH WAS REVERSED BY MIR-132-3P OVEREXPRESSION. MOREOVER, AUTOPHAGIC GENE EXPRESSION INDUCED BY MIR-132-3P KNOCKDOWN WAS REVERSED BY ATG16L1 KNOCKDOWN. BASED ON PRECIPITATION ASSAYS, LNCRNA UCA1 AND MIR-132-3P WERE SHOWN TO FORM A COMPLEX WITH THE TRANSCRIPTION FACTOR, EZH2, AND MIR-132-3P WAS SHOWN TO INTERACT WITH ATG16L1 BASED ON A LUCIFERASE ASSAY. FINALLY, LNCRNA UCA1 WAS SHOWN TO NEGATIVELY REGULATE MIR-132-3P EXPRESSION, AND MIR-132-3P WAS SHOWN TO NEGATIVELY REGULATE ATG16L1. SIGNIFICANCE: IN THIS CELL MODEL, LNCRNA UCA1 PROMOTES AUTOPHAGIC GENE EXPRESSION VIA EPIGENETIC REGULATION MEDIATED BY ATG16L1 AND MIR-132-3P. 2022