1 814 143 CHANGES IN SMALL NONCODING RNA EXPRESSION DURING CHONDROCYTE SENESCENCE. OBJECTIVE: OSTEOARTHRITIS (OA) IS CHARACTERIZED BY THE CHRONIC AND PROGRESSIVE DETERIORATION OF ARTICULAR CARTILAGE. CHONDROCYTE SENESCENCE COULD LEAD TO A SHIFT IN THE BALANCE BETWEEN EXTRACELLULAR MATRIX (ECM) COMPONENT SYNTHESIS AND DEGRADATION. SMALL NONCODING RNAS (SNCRNAS), INCLUDING MICRORNAS (MIRNAS), P-ELEMENT-INDUCED WIMPY TESTIS-(PIWI-) INTERACTING RNAS (PIRNAS), SMALL NUCLEOLAR RNAS (SNORNAS), SMALL NUCLEAR RNAS (SNRNAS), AND REPEAT-ASSOCIATED SIRNAS (RASIRNAS), ARE A CLASS OF IMPORTANT EPIGENETIC MOLECULES. WE AIMED TO GAIN INSIGHTS INTO THE CHANGES AND ROLES OF SNCRNA IN CHONDROCYTE SENESCENCE. DESIGN: HEALTHY MOUSE POSTNATAL CHONDROCYTES WERE ISOLATED, AND A REPLICATIVE AGING MODEL WAS CONSTRUCTED. WE USED SMALL RNA SEQUENCING (SMALL RNA-SEQ) TO GENERATE EXTENSIVE SMALL RNA DATA. WE IDENTIFIED DIFFERENTIALLY EXPRESSED SNCRNAS AND PERFORMED TISSUE-SPECIFIC ANALYSIS USING REAL-TIME QUANTITATIVE POLYMERASE CHAIN REACTION (QRT-PCR). BETA-GALACTOSIDASE STAINING WAS USED TO DETECT CHONDROCYTE SENESCENCE. THE RESULTS SHOWED THAT THE EXPRESSION PROFILES OF SNCRNA IN PASSAGE 5 CHONDROCYTES WERE SIGNIFICANTLY DIFFERENT FROM THOSE IN PASSAGE 0 CHONDROCYTES. THE EXPRESSION OF SNCRNA WAS TISSUE SPECIFIC. WE FOUND THAT 40 MIRNAS WERE UPREGULATED AND 70 MIRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT MIR-132-5P EXPRESSION INHIBITION PREVENTED CHONDROCYTE SENESCENCE. WE FOUND THAT 8 PIRNAS WERE UPREGULATED AND 17 PIRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT PIRNA PIR_025576 OVEREXPRESSION DELAYED CHONDROCYTE SENESCENCE. WE FOUND THAT 24 SNORNAS WERE UPREGULATED AND 28 SNORNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT SNORNA ENSMUSG00000087935 OVEREXPRESSION DELAYED CHONDROCYTE SENESCENCE. WE FOUND THAT 5 SNRNAS WERE UPREGULATED AND 6 SNRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT SNRNA ENSMUSG00000064682 OVEREXPRESSION DELAYED CHONDROCYTE SENESCENCE. WE FOUND THAT 1 RASIRNA WAS UPREGULATED AND 4 RASIRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE. CONCLUSIONS: THESE FINDINGS MIGHT PROVIDE NOVEL INSIGHTS INTO OA PATHOGENESIS AND CONTRIBUTE TO THE DEVELOPMENT OF CANDIDATES FOR TARGETED THERAPEUTICS IN OA. 2022 2 3947 27 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 3 2117 27 EPIGENETIC HISTONE METHYLATION MODULATES FIBROTIC GENE EXPRESSION. TGF-BETA1-INDUCED EXPRESSION OF EXTRACELLULAR MATRIX (ECM) GENES PLAYS A MAJOR ROLE IN THE DEVELOPMENT OF CHRONIC RENAL DISEASES SUCH AS DIABETIC NEPHROPATHY. ALTHOUGH MANY KEY TRANSCRIPTION FACTORS ARE KNOWN, MECHANISMS INVOLVING THE NUCLEAR CHROMATIN THAT MODULATE ECM GENE EXPRESSION REMAIN UNCLEAR. HERE, WE EXAMINED THE ROLE OF EPIGENETIC CHROMATIN MARKS SUCH AS HISTONE H3 LYSINE METHYLATION (H3KME) IN TGF-BETA1-INDUCED GENE EXPRESSION IN RAT MESANGIAL CELLS UNDER NORMAL AND HIGH-GLUCOSE (HG) CONDITIONS. TGF-BETA1 INCREASED THE EXPRESSION OF THE ECM-ASSOCIATED GENES CONNECTIVE TISSUE GROWTH FACTOR, COLLAGEN-ALPHA1[IOTA], AND PLASMINOGEN ACTIVATOR INHIBITOR-1. INCREASED LEVELS OF CHROMATIN MARKS ASSOCIATED WITH ACTIVE GENES (H3K4ME1, H3K4ME2, AND H3K4ME3), AND DECREASED LEVELS OF REPRESSIVE MARKS (H3K9ME2 AND H3K9ME3) AT THESE GENE PROMOTERS ACCOMPANIED THESE CHANGES IN EXPRESSION. TGF-BETA1 ALSO INCREASED EXPRESSION OF THE H3K4 METHYLTRANSFERASE SET7/9 AND RECRUITMENT TO THESE PROMOTERS. SET7/9 GENE SILENCING WITH SIRNAS SIGNIFICANTLY ATTENUATED TGF-BETA1-INDUCED ECM GENE EXPRESSION. FURTHERMORE, A TGF-BETA1 ANTIBODY NOT ONLY BLOCKED HG-INDUCED ECM GENE EXPRESSION BUT ALSO REVERSED HG-INDUCED CHANGES IN PROMOTER H3KME LEVELS AND SET7/9 OCCUPANCY. TAKEN TOGETHER, THESE RESULTS SHOW THE FUNCTIONAL ROLE OF EPIGENETIC CHROMATIN HISTONE H3KME IN TGF-BETA1-MEDIATED ECM GENE EXPRESSION IN MESANGIAL CELLS UNDER NORMAL AND HG CONDITIONS. PHARMACOLOGIC AND OTHER THERAPIES THAT REVERSE THESE MODIFICATIONS COULD HAVE POTENTIAL RENOPROTECTIVE EFFECTS FOR DIABETIC NEPHROPATHY. 2010 4 3935 37 LIVER-SPECIFIC KNOCKDOWN OF CLASS IIA HDACS HAS LIMITED EFFICACY ON GLUCOSE METABOLISM BUT ENTAILS SEVERE ORGAN SIDE EFFECTS IN MICE. HISTONE DEACETYLASES (HDACS) ARE IMPORTANT REGULATORS OF EPIGENETIC GENE MODIFICATION THAT ARE INVOLVED IN THE TRANSCRIPTIONAL CONTROL OF METABOLISM. IN PARTICULAR CLASS IIA HDACS HAVE BEEN SHOWN TO AFFECT HEPATIC GLUCONEOGENESIS AND PREVIOUS APPROACHES REVEALED THAT THEIR INHIBITION REDUCES BLOOD GLUCOSE IN TYPE 2 DIABETIC MICE. IN THE PRESENT STUDY, WE AIMED TO EVALUATE THE POTENTIAL OF CLASS IIA HDAC INHIBITION AS A THERAPEUTIC OPPORTUNITY FOR THE TREATMENT +OF METABOLIC DISEASES. FOR THAT, SIRNAS SELECTIVELY TARGETING HDAC4, 5 AND 7 WERE SELECTED AND USED TO ACHIEVE A COMBINATORIAL KNOCKDOWN OF THESE THREE CLASS IIA HDAC ISOFORMS. SUBSEQUENTLY, THE HEPATOCELLULAR EFFECTS AS WELL AS THE IMPACT ON GLUCOSE AND LIPID METABOLISM WERE ANALYZED IN VITRO AND IN VIVO. THE TRIPLE KNOCKDOWN RESULTED IN A STATISTICALLY SIGNIFICANT DECREASE OF GLUCONEOGENIC GENE EXPRESSION IN MURINE AND HUMAN HEPATOCYTE CELL MODELS. A SIMILAR HDAC-INDUCED DOWNREGULATION OF HEPATIC GLUCONEOGENESIS GENES COULD BE ACHIEVED IN MICE USING A LIVER-SPECIFIC LIPID NANOPARTICLE SIRNA FORMULATION. HOWEVER, THE EFFICACY ON WHOLE BODY GLUCOSE METABOLISM ASSESSED BY PYRUVATE-TOLERANCE TESTS WERE ONLY LIMITED AND DID NOT OUTWEIGH THE SAFETY FINDINGS OBSERVED BY HISTOPATHOLOGICAL ANALYSIS IN SPLEEN AND KIDNEY. MECHANISTICALLY, AFFYMETRIX GENE EXPRESSION STUDIES PROVIDE EVIDENCE THAT CLASS IIA HDACS DIRECTLY TARGET OTHER KEY FACTORS BEYOND THE DESCRIBED FORKHEAD BOX (FOXP) TRANSCRIPTION REGULATORS, SUCH AS HEPATOCYTE NUCLEAR FACTOR 4 ALPHA (HNF4A). DOWNSTREAM OF THESE FACTORS SEVERAL ADDITIONAL PATHWAYS WERE REGULATED NOT MERELY INCLUDING GLUCOSE AND LIPID METABOLISM AND TRANSPORT. IN CONCLUSION, THE LIVER-DIRECTED COMBINATORIAL KNOCKDOWN OF HDAC4, 5 AND 7 BY THERAPEUTIC SIRNAS AFFECTED MULTIPLE PATHWAYS IN VITRO, LEADING IN VIVO TO THE DOWNREGULATION OF GENES INVOLVED IN GLUCONEOGENESIS. HOWEVER, THE EFFECTS ON GENE EXPRESSION LEVEL WERE NOT PARALLELED BY A SIGNIFICANT REDUCTION OF GLUCONEOGENESIS IN MICE. COMBINED KNOCKDOWN OF HDAC ISOFORMS WAS ASSOCIATED WITH SEVERE ADVERSE EFFECTS IN VIVO, CHALLENGING THIS APPROACH AS A TREATMENT OPTION FOR CHRONIC METABOLIC DISORDERS LIKE TYPE 2 DIABETES. 2020 5 412 37 ANALYSIS OF LONG NON-CODING RNA (LNCRNA) EXPRESSION IN HEPATITIS B PATIENTS. LONG NON-CODING RNAS (LNCRNAS) HAVE BEEN IMPLICATED IN NUMEROUS BIOLOGICAL PROCESSES, INCLUDING EPIGENETIC REGULATION, CELL-CYCLE CONTROL, AND TRANSCRIPTIONAL/TRANSLATIONAL REGULATION OF GENE EXPRESSION. DIFFERENTIAL EXPRESSION OF LNCRNAS AND DISRUPTION OF THE REGULATORY PROCESSES ARE RECOGNIZED AS CRITICAL STEPS IN CANCER DEVELOPMENT. THE ROLE OF LNCRNAS IN HEPATITIS B VIRUS (HBV) INFECTION IS NOT WELL UNDERSTOOD. HERE WE ANALYZED THE EXPRESSION OF 135 LNCRNAS IN PLASMA SAMPLES OF 82 HBV PATIENTS (CLASSIFIED AS CHRONIC PATIENTS, INACTIVE CARRIERS, OR RESOLVED PATIENTS) AT DIAGNOSIS AND AT 12 MONTHS OF TREATMENT IN RELATION TO CONTROL GROUP (81 HEALTHY VOLUNTEERS). WE ALSO INVESTIGATED THE EFFECT OF SMALL INTERFERING RNA (SIRNA)-MEDIATED SILENCING OF LINCRNA-SFMBT2 ON HBV-POSITIVE HUMAN LIVER CANCER CELL LINE. LNCRNA EXPRESSION WAS ANALYZED BY QUANTITATIVE REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION (QRT-PCR). CHEMICALLY SYNTHESIZED SIRNAS WERE TRANSFECTED INTO THE CELL LINES USING LIPOFECTAMINE 2000 REAGENT (THERMO FISHER SCIENTIFIC). HBV DNA AND HBSAG AND HBEAG WERE DETECTED IN TRANSFECTED CULTURES BY REAL-TIME PCR AND ELISA, RESPECTIVELY, USING COMMERCIAL KITS. WE OBSERVED CHANGES IN LNCRNA EXPRESSION IN ALL THREE HBV GROUPS, COMPARED TO CONTROL GROUP. MOST NOTABLY, THE EXPRESSION OF ANTI-NOS2A, LINCRNA-SFMBT2, AND ZFHX2AS WAS SIGNIFICANTLY INCREASED AND EXPRESSION OF Y5 LNCRNA WAS DECREASED IN CHRONIC HBV PATIENTS. A DECREASED Y5 EXPRESSION AND INCREASED LINCRNA-SFMBT2 EXPRESSION WERE OBSERVED IN INACTIVE HBSAG CARRIERS. THE EXPRESSION OF HOTTIP, MEG9, AND PCAT-32 WAS INCREASED IN RESOLVED HBV PATIENTS, AND NO SIGNIFICANT CHANGE IN THE EXPRESSION OF Y5 WAS OBSERVED, COMPARED TO CONTROL GROUP. SIRNA-MEDIATED INHIBITION OF LINCRNA-SFMBT2 DECREASED THE LEVEL OF HBV DNA IN HUMAN LIVER CANCER CELLS. FURTHER RESEARCH IS NEEDED TO CONFIRM THE PROGNOSTIC AS WELL AS THERAPEUTIC ROLE OF THESE LNCRNAS IN HBV PATIENTS. 2018 6 3904 31 LEPROSY PIRNOME: EXPLORING NEW POSSIBILITIES FOR AN OLD DISEASE. LEPROSY, WHICH IS CAUSED BY THE HUMAN PATHOGEN MYCOBACTERIUM LEPRAE, CAUSES NERVE DAMAGE, DEFORMITY AND DISABILITY IN OVER 200,000 PEOPLE EVERY YEAR. BECAUSE OF THE LONG DOUBLING TIME OF M. LEPRAE (13 DAYS) AND THE DELAYED ONSET OF DETECTABLE SYMPTOMS, WHICH IS ESTIMATED TO BE APPROXIMATELY 3-7 YEARS AFTER INFECTION, THERE IS ALWAYS A LARGE PERCENTAGE OF SUBCLINICALLY INFECTED INDIVIDUALS IN THE POPULATION WHO WILL EVENTUALLY DEVELOP THE DISEASE, MAINLY IN ENDEMIC COUNTRIES. PIRNAS COMPRISE THE LARGEST GROUP OF SMALL NONCODING RNAS FOUND IN HUMANS, AND THEY ARE DISTINCT FROM MICRORNAS (MIRNAS) AND SMALL INTERFERING RNAS (SIRNAS). PIRNAS FUNCTION IN TRANSPOSON SILENCING, EPIGENETIC REGULATION, AND GERMLINE DEVELOPMENT. THE FUNCTIONAL ROLE OF PIRNAS AND THEIR ASSOCIATED PIWI PROTEINS HAVE STARTED TO EMERGE IN THE DEVELOPMENT OF HUMAN CANCERS AND VIRAL INFECTIONS, BUT THEIR RELEVANCE TO BACTERIAL DISEASES HAS NOT BEEN INVESTIGATED. THE PRESENT STUDY REPORTS THE PIRNOME OF HUMAN SKIN, REVEALING THAT ALL BUT ONE OF THE PIRNAS EXAMINED ARE DOWNREGULATED IN LEPROSY SKIN LESIONS. CONSIDERING THAT ONE OF THE BEST CHARACTERIZED FUNCTIONS OF PIRNAS IN HUMANS IS POSTTRANSCRIPTIONAL MRNA SILENCING, THEIR FUNCTIONS ARE SIMILAR TO WHAT WE HAVE DESCRIBED FOR MIRNAS, INCLUDING ACTING ON APOPTOSIS, M. LEPRAE RECOGNITION AND ENGULFMENT, SCHWANN CELL (SC) DEMYELINATION, EPITHELIAL-MESENCHYMAL TRANSITION (EMT), LOSS OF SENSATION AND NEUROPATHIC PAIN. IN ADDITION TO NEW FINDINGS ON LEPROSY PHYSIOPATHOLOGY, THE DISCOVERY OF RELEVANT PIRNAS INVOLVED IN DISEASE PROCESSES IN HUMAN SKIN MAY PROVIDE NEW CLUES FOR THERAPEUTIC TARGETS, SPECIFICALLY TO CONTROL NERVE DAMAGE, A PROMINENT FEATURE OF LEPROSY THAT HAS NO CURRENTLY AVAILABLE PHARMACEUTICAL TREATMENT. 2020 7 4364 31 MIRNA DEREGULATION BY EPIGENETIC SILENCING DISRUPTS SUPPRESSION OF THE ONCOGENE PLAG1 IN CHRONIC LYMPHOCYTIC LEUKEMIA. MICRORNAS (MIRNA) PLAY A KEY ROLE IN CELLULAR REGULATION AND, IF DEREGULATED, IN THE DEVELOPMENT OF NEOPLASTIC DISORDERS INCLUDING CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). RNAS FROM PRIMARY CELLS OF 50 TREATMENT-NAIVE CLL PATIENTS AND PERIPHERAL B CELLS OF 14 HEALTHY DONORS WERE APPLIED TO MIRNA EXPRESSION PROFILING USING BEAD CHIP TECHNOLOGY. IN CLL CELLS, A SET OF 7 UP- AND 19 DOWN-REGULATED MIRNAS WAS IDENTIFIED. AMONG THE MIRNAS DOWN-REGULATED IN CLL CELLS, 6 OF 10 MIRNA PROMOTERS EXAMINED SHOWED GAIN OF METHYLATION COMPARED WITH NORMAL B-CELL CONTROLS. SUBSEQUENT TARGET PREDICTION OF DEREGULATED MIRNAS REVEALED A HIGHLY SIGNIFICANT BINDING PREDICTION AT THE 3' UNTRANSLATED REGION OF THE PLEOMORPHIC ADENOMA GENE 1 (PLAG1) ONCOGENE. LUCIFERASE REPORTER ASSAYS INCLUDING SITE-DIRECTED MUTAGENESIS OF BINDING SITES REVEALED A SIGNIFICANT REGULATION OF PLAG1 BY MIR-181A, MIR-181B, MIR-107, AND MIR-424. ALTHOUGH EXPRESSION OF PLAG1 MRNA WAS NOT AFFECTED, PLAG1 PROTEIN EXPRESSION WAS SHOWN TO BE SIGNIFICANTLY ELEVATED IN CLL CELLS COMPARED WITH THE LEVELS IN HEALTHY DONOR B CELLS. IN SUMMARY, WE COULD DEMONSTRATE DISRUPTION OF MIRNA-MEDIATED TRANSLATIONAL CONTROL, PARTLY DUE TO EPIGENETIC TRANSCRIPTIONAL SILENCING OF MIRNAS, WITH SUBSEQUENT OVEREXPRESSION OF THE ONCOGENIC TRANSCRIPTION FACTOR PLAG1 AS A PUTATIVE NOVEL MECHANISM OF CLL PATHOGENESIS. 2009 8 6235 30 THE M(6)A DEMETHYLASE FTO PROMOTES RENAL EPITHELIAL-MESENCHYMAL TRANSITION BY REDUCING THE M(6)A MODIFICATION OF LNCRNA GAS5. BACKGROUND: RENAL INTERSTITIAL FIBROSIS (RIF) IS THE MAIN PATHOLOGICAL CHANGE OF A VARIETY OF CHRONIC KIDNEY DISEASES (CKD). EPIGENETIC MODIFICATIONS OF FIBROSIS-PRONE GENES REGULATE RIF PROGRESSION. THIS STUDY AIMED TO INVESTIGATE LONG NON-CODING RNA (LNCRNA) N6-METHYLADENOSINE (M(6)A) MODIFICATION AND ITS ROLE IN REGULATING RIF PROGRESSION. METHODS: UNILATERAL URETERAL OCCLUSION (UUO) WAS EMPLOYED TO CONSTRUCT THE RIF IN VIVO MODEL; AND TGF-BETA1-TREATED HK-2 AND HKC-8 CELLS WERE USED FOR IN VITRO EXPERIMENTS. THE MRNA AND PROTEIN EXPRESSIONS WERE ASSESSED USING QRT-PCR AND WESTERN BLOT. THE PROLIFERATION AND MIGRATION WERE EVALUATED BY EDU ASSAY AND TRANSWELL ASSAY, RESPECTIVELY. IN ADDITION, LEVELS OF INFLAMMATORY CYTOKINES WERE DETERMINED BY ELISA ASSAY AND QRT-PCR. MOREOVER, LNCRNA GAS5 M(6)A LEVEL WAS DETECTED USING ME-RIP ASSAY. HE AND MASSON STAINING WERE EMPLOYED TO EVALUATE FIBROTIC LESIONS OF THE KIDNEY. RESULTS: FTO EXPRESSION WAS ELEVATED IN HK-2 AND HKC-8 CELLS AFTER TGF-BETA1 TREATMENT AND MOUSE KIDNEY TISSUE FOLLOWING UUO, AND LNCRNA GAS5 WAS DOWNREGULATED. LNCRNA GAS5 OVEREXPRESSION OR FTO SILENCING SUPPRESSED TGF-BETA1-INDUCED THE INCREASE OF EMT-RELATED PROTEINS (VIMENTIN, SNAIL AND N-CADHERIN) AND INFLAMMATORY CYTOKINES (IL-6, IL-1BETA AND TNF-ALPHA) LEVELS IN HK-2 CELLS. FTO SUPPRESSED LNCRNA GAS5 EXPRESSION BY REDUCING THE M6A MODIFICATION OF LNCRNA GAS5. ADDITIONALLY, FTO KNOCKDOWN COULD SUPPRESS EMT PROCESS AND INFLAMMATION RESPONSE INDUCED BY TGF-BETA1 AND UUO IN VITRO AND IN VIVO. AS EXPECTED, FTO KNOCKDOWN ABROGATED THE PROMOTION EFFECTS OF LNCRNA GAS5 SILENCING ON TGF-BETA1-INDUCED EMT PROCESS AND INFLAMMATION RESPONSE IN HK-2 AND HKC-8 CELLS. CONCLUSION: FTO PROMOTED EMT PROCESS AND INFLAMMATION RESPONSE THROUGH REDUCING THE M(6)A MODIFICATION OF LNCRNA GAS5. 2022 9 3721 34 INHIBITION OF CLASS II HISTONE DEACETYLASES IN THE SPINAL CORD ATTENUATES INFLAMMATORY HYPERALGESIA. BACKGROUND: SEVERAL CLASSES OF HISTONE DEACETYLASES (HDACS) ARE EXPRESSED IN THE SPINAL CORD THAT IS A CRITICAL STRUCTURE OF THE NOCICEPTIVE PATHWAY. HDAC-REGULATED HISTONE ACETYLATION IS AN IMPORTANT COMPONENT OF CHROMATIN REMODELING LEADING TO EPIGENETIC REGULATION OF GENE TRANSCRIPTION. TO UNDERSTAND THE ROLE OF HISTONE ACETYLATION IN EPIGENETIC REGULATION OF PATHOLOGICAL PAIN, WE HAVE STUDIED THE IMPACT OF DIFFERENT CLASSES OF HDACS IN THE SPINAL CORD ON INFLAMMATORY HYPERALGESIA INDUCED BY COMPLETE FREUND'S ADJUVANT (CFA). RESULTS: WE INTRATHECALLY APPLIED INHIBITORS SPECIFIC TO DIFFERENT CLASSES OF HDACS AND EVALUATED THEIR IMPACT ON INFLAMMATORY HYPERALGESIA. PRE-INJECTED INHIBITORS TARGETING CLASS I AS WELL AS II (SAHA, TSA, LAQ824) OR IIA (VPA, 4-PB) HDACS SIGNIFICANTLY DELAYED THE THERMAL HYPERALGESIA INDUCED BY UNILATERAL CFA INJECTION IN THE HINDPAW. EXISTING HYPERALGESIA INDUCED BY CFA WAS ALSO ATTENUATED BY THE HDAC INHIBITORS (HDACIS). IN CONTRAST, THESE INHIBITORS DID NOT INTERFERE WITH THE THERMAL RESPONSE EITHER IN NAIVE ANIMALS, OR ON THE CONTRALATERAL SIDE OF INFLAMED ANIMALS. INTERESTINGLY, MS-275 THAT SPECIFICALLY INHIBITS CLASS I HDACS FAILED TO ALTER THE HYPERALGESIA ALTHOUGH IT INCREASED HISTONE 3 ACETYLATION IN THE SPINAL CORD AS SAHA DID. USING IMMUNOBLOT ANALYSIS, WE FURTHER FOUND THAT THE LEVELS OF CLASS IIA HDAC MEMBERS (HDAC4, 5, 7, 9) IN THE SPINAL DORSAL HORN WERE UPREGULATED FOLLOWING CFA INJECTION WHILE THOSE OF CLASS I HDAC MEMBERS (HDAC1, 2, 3) REMAINED STABLE OR WERE SLIGHTLY REDUCED. CONCLUSIONS: OUR DATA SUGGEST THAT ACTIVITY OF CLASS II HDACS IN THE SPINAL CORD IS CRITICAL TO THE INDUCTION AND MAINTENANCE OF INFLAMMATORY HYPERALGESIA INDUCED BY CFA, WHILE ACTIVITY OF CLASS I HDACS MAY BE UNNECESSARY. COMPARISON OF THE EFFECTS OF HDACIS SPECIFIC TO CLASS II AND IIA AS WELL AS THE EXPRESSION PATTERN OF DIFFERENT HDACS IN THE SPINAL CORD IN RESPONSE TO CFA SUGGESTS THAT THE MEMBERS OF CLASS IIA HDACS MAY BE POTENTIAL TARGETS FOR ATTENUATING PERSISTENT INFLAMMATORY PAIN. 2010 10 3358 23 HISTONE H3K9 METHYLATION REGULATES CHRONIC STRESS AND IL-6-INDUCED COLON EPITHELIAL PERMEABILITY AND VISCERAL PAIN. BACKGROUND: CHRONIC STRESS IS ASSOCIATED WITH ACTIVATION OF THE HPA AXIS, ELEVATION IN PRO-INFLAMMATORY CYTOKINES, DECREASE IN INTESTINAL EPITHELIAL CELL TIGHT JUNCTION (TJ) PROTEINS, AND ENHANCED VISCERAL PAIN. IT IS UNKNOWN WHETHER EPIGENETIC REGULATORY PATHWAYS PLAY A ROLE IN CHRONIC STRESS-INDUCED INTESTINAL BARRIER DYSFUNCTION AND VISCERAL HYPERALGESIA. METHODS: YOUNG ADULT MALE RATS WERE SUBJECTED TO WATER AVOIDANCE STRESS +/- H3K9 METHYLATION INHIBITORS OR SIRNAS. VISCERAL PAIN RESPONSE WAS ASSESSED. DIFFERENTIATED CACO-2/BBE CELLS AND HUMAN COLONOIDS WERE TREATED WITH CORTISOL OR IL-6 +/- ANTAGONISTS. EXPRESSION OF TJ, IL-6, AND H3K9 METHYLATION STATUS AT GENE PROMOTERS WAS MEASURED. TRANSEPITHELIAL ELECTRICAL RESISTANCE AND FITC-DEXTRAN PERMEABILITY WERE EVALUATED. KEY RESULTS: CHRONIC STRESS INDUCED IL-6 UP-REGULATION PRIOR TO A DECREASE IN TJ PROTEINS IN THE RAT COLON. THE IL-6 LEVEL INVERSELY CORRELATED WITH OCCLUDIN EXPRESSION. TREATMENT WITH IL-6 DECREASED OCCLUDIN AND INDUCED VISCERAL HYPERALGESIA. CHRONIC STRESS AND IL-6 INCREASED H3K9 METHYLATION AND DECREASED TRANSCRIPTIONAL GR BINDING TO THE OCCLUDIN GENE PROMOTER, LEADING TO DOWN-REGULATION OF PROTEIN EXPRESSION AND INCREASE IN PARACELLULAR PERMEABILITY. INTRARECTAL ADMINISTRATION OF A H3K9 METHYLATION ANTAGONIST PREVENTED CHRONIC STRESS-INDUCED VISCERAL HYPERALGESIA IN THE RAT. IN A HUMAN COLONOID MODEL, CORTISOL DECREASED OCCLUDIN EXPRESSION, WHICH WAS PREVENTED BY THE GR ANTAGONIST RU486, AND IL-6 INCREASED H3K9 METHYLATION AND DECREASED TJ PROTEIN LEVELS, WHICH WERE PREVENTED BY INHIBITORS OF H3K9 METHYLATION. CONCLUSIONS & INFERENCES: OUR FINDINGS SUPPORT A NOVEL ROLE FOR METHYLATION OF THE REPRESSIVE HISTONE H3K9 TO REGULATE CHRONIC STRESS, PRO-INFLAMMATORY CYTOKINE-MEDIATED REDUCTION IN COLON TJ PROTEIN LEVELS, AND INCREASE IN PARACELLULAR PERMEABILITY AND VISCERAL HYPERALGESIA. 2020 11 2326 37 EPIGENETIC REGULATION OF HOTAIR IN ADVANCED CHRONIC MYELOID LEUKEMIA. PURPOSE: CHRONIC MYELOID LEUKEMIA (CML) ACCOUNTS FOR ~10% OF LEUKEMIA CASES, AND ITS PROGRESSION INVOLVES EPIGENETIC GENE REGULATION. THIS STUDY INVESTIGATED EPIGENETIC REGULATION OF HOTAIR AND ITS TARGET MICRORNA, MIR-143, IN ADVANCED CML. PATIENTS AND METHODS: WE FIRST ISOLATED BONE MARROW MONONUCLEAR CELLS FROM 70 PATIENTS WITH DIFFERENT PHASES OF CML AND FROM HEALTHY DONORS AS NORMAL CONTROL; WE ALSO CULTURED K562 AND KCL22 CELLS, TREATED WITH DEMETHYLATION DRUG; MTT ASSAY, FLOW CYTOMETRY, QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QPCR), METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP), WESTERN BLOT, LUCIFERASE ASSAY, RNA PULL-DOWN ASSAY AND RNA-BINDING PROTEIN IMMUNOPRECIPITATION (RIP) ASSAY WERE PERFORMED. RESULT: AS MEASURED BY QPCR, HOTAIR EXPRESSION IN K562 CELLS, KCL22 CELLS, AND SAMPLES FROM CASES OF ADVANCED-STAGE CML INCREASED WITH LEVELS OF SEVERAL DNA METHYLTRANSFERASES AND HISTONE DEACETYLATES, INCLUDING DNMT1, DNMT3A, HDAC1, EZH2, AND LSD1, AND MIR-143 LEVELS WERE DECREASED AND HOTAIR LEVELS WERE INCREASED. TREATMENT WITH 5-AZACYTIDINE, A DNA METHYLATION INHIBITOR, DECREASED DNMT1, DNMT3A, HDAC1, EZH2, LSD1 MRNA, PROTEIN LEVELS, AND HOTAIR MRNA LEVELS BUT INCREASED MIR-143 LEVELS. HOTAIR KNOCKDOWN AND MIR-143 OVEREXPRESSION BOTH INHIBITED PROLIFERATION AND PROMOTED APOPTOSIS IN KCL22 AND K562 CELLS THROUGH THE PI3K/AKT PATHWAY. RNA PULL-DOWN, MASS SPECTROMETRY, AND RIP ASSAYS SHOWED THAT HOTAIR INTERACTED WITH EZH2 AND LSD1. A DUAL-LUCIFERASE ASSAY DEMONSTRATED THAT HOTAIR INTERACTED WITH MIR-143. CONCLUSION: OUR FINDINGS DEMONSTRATE THE KEY EPIGENETIC INTERACTIONS OF HOTAIR RELATED TO CML PROGRESSION AND SUGGEST HOTAIR AS A POTENTIAL THERAPEUTIC TARGET FOR ADVANCED CML. FURTHERMORE, OUR RESULTS SUPPORT THE USE OF DEMETHYLATION DRUGS AS A CML TREATMENT STRATEGY. 2018 12 3347 23 HISTONE DEACETYLASES MEDIATE THE SILENCING OF MIR-15A, MIR-16, AND MIR-29B IN CHRONIC LYMPHOCYTIC LEUKEMIA. CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) DEMONSTRATES A GLOBAL DOWN-REGULATION OF MIR-15A AND MIR-16 AND A SELECTIVE SILENCING OF THE RELATED MIR-29B IN AGGRESSIVE DISEASE. DELETIONS IN CHROMOSOME 13 [DEL(13Q14)] PARTIALLY ACCOUNT FOR THE LOSS OF EXPRESSION OF MIR-15A AND MIR-16, BUT THE MECHANISMS BY WHICH MIR-29B BECOMES SILENCED IS UNKNOWN. IN THE PRESENT STUDY, WE SHOW THAT THE HISTONE DEACETYLASES (HDACS) ARE OVEREXPRESSED IN CLL AND MEDIATE THE EPIGENETIC SILENCING OF MIR-15A, MIR-16, AND MIR-29B. HDAC INHIBITION TRIGGERED THE ACCUMULATION OF THE TRANSCRIPTIONALLY ACTIVATING CHROMATIN MODIFICATION H3K4ME2 AND RESTORED THE EXPRESSION OF MIR-15A, MIR-16, AND MIR-29B IN APPROXIMATELY 35% OF SAMPLES. ECTOPIC EXPRESSION OF MIR-15A AND MIR-16 AND HDAC INHIBITION-INDUCED EXPRESSION OF MIR-15A, MIR-16, OR MIR-29B IN PRIMARY CLL CELLS WAS ASSOCIATED WITH DECLINES IN THE LEVELS OF MCL-1, BUT NOT BCL-2, MITOCHONDRIAL DYSFUNCTION, AND INDUCTION OF CELL DEATH. THEREFORE, OUR RESULTS SHOW THAT HDACS ABERRANTLY SILENCE THE EXPRESSION OF THE CRITICAL TUMOR SUPPRESSORS MIR-15A, MIR-16, AND MIR-29B IN CLL. DEACETYLASE INHIBITION MAY BE A THERAPEUTIC STRATEGY THAT RESTORES THE EXPRESSION OF THESE MIRS TO ANTAGONIZE MCL-1, AN IMPORTANT SURVIVAL PROTEIN IN THESE CELLS. CONSEQUENTLY, CLL PATIENTS WHO EXHIBIT SUCH EPIGENETIC SILENCING MAY BENEFIT FROM HDAC INHIBITOR-BASED THERAPY. 2012 13 1826 42 EFFECTS OF HISTONE DEACETYLASE INHIBITOR ON EXTRACELLULAR MATRIX PRODUCTION IN HUMAN NASAL POLYP ORGAN CULTURES. BACKGROUND: NASAL POLYPOSIS IS ASSOCIATED WITH A CHRONIC INFLAMMATORY CONDITION OF THE SINONASAL MUCOSA AND INVOLVES MYOFIBROBLAST DIFFERENTIATION AND EXTRACELLULAR MATRIX (ECM) ACCUMULATION. EPIGENETIC MODULATION BY HISTONE DEACETYLASE (HDAC) INHIBITORS INCLUDING TRICHOSTATIN A (TSA) HAS BEEN REPORTED TO HAVE INHIBITORY EFFECTS ON MYOFIBROBLAST DIFFERENTIATION IN LUNG AND RENAL FIBROBLASTS. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE THE INHIBITORY EFFECT OF TSA ON MYOFIBROBLAST DIFFERENTIATION AND ECM PRODUCTION IN NASAL POLYP ORGAN CULTURES. METHODS: NASAL POLYP TISSUES FROM 18 PATIENTS WERE ACQUIRED DURING ENDOSCOPIC SINUS SURGERY. AFTER ORGAN CULTURE, NASAL POLYPS WERE STIMULATED WITH TGF-BETA1 AND THEN TREATED WITH TSA. ALPHA-SMOOTH MUSCLE ACTIN (ALPHA-SMA), FIBRONECTIN, AND COLLAGEN TYPE I EXPRESSION LEVELS WERE EXAMINED BY REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION (PCR), REAL-TIME PCR, WESTERN BLOT, AND IMMUNOFLUORESCENT STAINING. HDAC2, HDAC4, AND ACETYLATED H4 EXPRESSION LEVELS WERE ASSAYED BY WESTERN BLOT. CYTOTOXICITY WAS ANALYZED BY THE TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE BIOTIN-DUTP NICK END LABELING ASSAY. RESULTS: THE EXPRESSION LEVELS OF ALPHA-SMA, FIBRONECTIN, AND COLLAGEN TYPE 1 WERE INCREASED IN NASAL POLYP AFTER TRANSFORMING GROWTH FACTOR (TGF) BETA1 TREATMENT. TSA-INHIBITED TGF-BETA1 INDUCED THESE GENE AND PROTEIN EXPRESSION LEVELS. FURTHERMORE, TSA SUPPRESSED PROTEIN EXPRESSION LEVELS OF HDAC2 AND HDAC4. HOWEVER, TSA INDUCED HYPERACETYLATION OF HISTONES H4. TREATMENT WITH TGF-BETA1 WITH OR WITHOUT TSA DID NOT HAVE CYTOTOXIC EFFECT. CONCLUSION: THESE FINDINGS PROVIDE NOVEL INSIGHTS INTO THE EPIGENETIC REGULATION IN MYOFIBROBLAST DIFFERENTIATION AND ECM PRODUCTION OF NASAL POLYP. TSA COULD BE A CANDIDATE OF A THERAPEUTIC AGENT FOR REVERSING THE TGF-BETA1-INDUCED ECM SYNTHESIS THAT LEADS TO NASAL POLYP DEVELOPMENT. 2013 14 5850 28 SUBEROYLANILIDE HYDROXAMIC ACID (SAHA) REDUCES FIBROSIS MARKERS AND DEACTIVATES HUMAN STELLATE CELLS VIA THE EPITHELIAL-MESENCHYMAL TRANSITION (EMT). HEPATIC FIBROSIS IS KNOWN AS THE ACCUMULATION OF CONNECTIVE TISSUE SECONDARY TO CHRONIC DAMAGE TO THE LIVER. EPITHELIAL-MESENCHYMAL TRANSITION (EMT) CORRESPONDING INCREASE IN LIVER FIBROGENESIS WAS SHOWN WITH IMMUNOHISTOCHEMISTRY AND PCR-BASED STUDIES. SUBEROYLANILIDE HYDROXAMIC ACID (SAHA), A SYNTHETIC COMPOUND APPROVED AS A HISTONE DEACETYLASE INHIBITOR (HDAC) BY THE FDA TO TREAT CUTANEOUS T-CELL LYMPHOMA IS UNDER INVESTIGATION FOR THE TREATMENT OF LUNG AND RENAL FIBROSIS. EXPERIMENTAL MODELING FOR HEPATIC FIBROSIS CAN BE CONSTRUCTED WITH AN LX2 CELL LINE ISOLATED FROM HUMAN HEPATIC STELLATE CELLS (HSCS). IN THIS STUDY, WE AIMED TO INVESTIGATE THE MODULATION OF SAHA IN THE PATHOGENESIS OF LIVER FIBROSIS BY DETECTING THE LEVELS OF PROTEINS; (E-CADHERIN (E-CAD), N-CADHERIN (N-CAD), VIMENTIN (VIM), AND GENES; E-CAD, N-CAD, VIM, TRANSFORMING GROWTH FACTOR-BETA (TGF-BETA), ALPHA-SMOOTH MUSCLE ACTIN (ALPHA-SMA), TYPE 1 COLLAGEN (COL1A1), TYPE 3 COLLAGEN (COL3A1)) THAT PLAY A SIGNIFICANT ROLE IN EMT WITH THE LX2 CELL LINE. WE ALSO EVALUATED THE ACTION OF SAHA WITH CELL PROLIFERATION, CLONOGENIC, AND MIGRATION ASSAY. CELL PROLIFERATION WAS PERFORMED BY FLOW CYTOMETRY. ALL THE PROTEIN LEVELS WERE DETERMINED BY WESTERN BLOT ANALYSIS, AND GENE EXPRESSION LEVELS WERE MEASURED BY REAL-TIME PCR. OUR STUDY OBSERVED THAT SAHA TREATMENT DECREASED CELL VIABILITY, COLONY FORMATION AND MIGRATION IN LX2 CELLS. WE FOUND THAT SAHA INCREASED E-CAD EXPRESSION LEVEL, WHILE IT DECREASED N-CAD, VIM, COL1A1, COL3A1, ALPHA-SMA TGF-BETA GENES EXPRESSION LEVELS. SAHA DECREASED THE LEVEL OF E-CAD, N-CAD, AND VIM PROTEIN LEVELS. WE THOUGHT THAT SAHA POSSESSES POTENT ANTIFIBROTIC AND ANTI-EMT PROPERTIES IN LX2. 2021 15 5120 29 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 16 699 34 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 17 598 23 BETA-ADRENERGIC SIGNALING PROMOTES TUMOR ANGIOGENESIS AND PROSTATE CANCER PROGRESSION THROUGH HDAC2-MEDIATED SUPPRESSION OF THROMBOSPONDIN-1. CHRONIC BEHAVIORAL STRESS AND BETA-ADRENERGIC SIGNALING HAVE BEEN SHOWN TO PROMOTE CANCER PROGRESSION, WHOSE UNDERLYING MECHANISMS ARE LARGELY UNCLEAR, ESPECIALLY THE INVOLVEMENT OF EPIGENETIC REGULATION. HISTONE DEACETYLASE-2 (HDAC2), AN EPIGENETIC REGULATOR, IS CRITICAL FOR STRESS-INDUCED CARDIAC HYPERTROPHY. IT IS UNKNOWN WHETHER IT IS NECESSARY FOR BETA-ADRENERGIC SIGNALING-PROMOTED CANCER PROGRESSION. USING XENOGRAFT MODELS, WE SHOWED THAT CHRONIC BEHAVIORAL STRESS AND BETA-ADRENERGIC SIGNALING PROMOTE ANGIOGENESIS AND PROSTATE CANCER PROGRESSION. HDAC2 WAS INDUCED BY BETA-ADRENERGIC SIGNALING IN VITRO AND IN MOUSE XENOGRAFTS. WE NEXT UNCOVERED THAT HDAC2 IS A DIRECT TARGET OF CAMP RESPONSE ELEMENT-BINDING PROTEIN (CREB) THAT IS ACTIVATED BY BETA-ADRENERGIC SIGNALING. NOTABLY, HDAC2 IS NECESSARY FOR BETA-ADRENERGIC SIGNALING TO INDUCE ANGIOGENESIS. WE FURTHER DEMONSTRATED THAT, UPON CREB ACTIVATION, HDAC2 REPRESSES THROMBOSPONDIN-1 (TSP1), A POTENT ANGIOGENESIS INHIBITOR, THROUGH EPIGENETIC REGULATION. TOGETHER, THESE DATA ESTABLISH A NOVEL PATHWAY THAT HDAC2 AND TSP1 ACT DOWNSTREAM OF CREB ACTIVATION IN BETA-ADRENERGIC SIGNALING TO PROMOTE CANCER PROGRESSION. 2017 18 1654 24 DORSAL ROOT GANGLIA COACTIVATOR-ASSOCIATED ARGININE METHYLTRANSFERASE 1 CONTRIBUTES TO PERIPHERAL NERVE INJURY-INDUCED PAIN HYPERSENSITIVITIES. NEUROPATHIC PAIN IS ASSOCIATED WITH GENE EXPRESSION CHANGES WITHIN THE DORSAL ROOT GANGLION (DRG) AFTER PERIPHERAL NERVE INJURY, WHICH INVOLVES EPIGENETIC MECHANISMS. COACTIVATOR-ASSOCIATED ARGININE METHYLTRANSFERASE 1 (CARM1), AN EPIGENETIC ACTIVATOR, REGULATES GENE TRANSCRIPTIONAL ACTIVITY BY PROTEIN POSTTRANSLATIONAL MODIFICATIONS. HOWEVER, WHETHER CARM1 PLAYS AN ESSENTIAL ROLE IN THE DEVELOPMENT AND MAINTENANCE OF NEUROPATHIC PAIN IS UNKNOWN. WE REPORT HERE THAT PERIPHERAL NERVE INJURY INDUCED THE UPREGULATION OF THE MRNA AND PROTEIN EXPRESSION OF CARM1 IN THE INJURED DRG, AND BLOCKING ITS EXPRESSION THROUGH SMALL INTERFERING RNA (SIRNA) IN THE INJURED DRG ATTENUATED THE DEVELOPMENT AND MAINTENANCE OF NEUROPATHIC PAIN. FURTHERMORE, PHARMACOLOGICAL INHIBITION OF CARM1 MITIGATED PERIPHERAL NERVE INJURY-INDUCED MECHANICAL ALLODYNIA AND THERMAL HYPERALGESIA. GIVEN THAT CARM1 INHIBITION OR KNOCKDOWN ATTENUATED THE INDUCTION AND MAINTENANCE OF NEUROPATHIC PAIN AFTER PERIPHERAL NERVE INJURY, OUR FINDINGS SUGGEST THAT CARM1 MAY SERVE AS A PROMISING THERAPEUTIC TARGET FOR NEUROPATHIC PAIN TREATMENT IN CLINICAL APPLICATIONS. 2018 19 6658 36 UPREGULATED LNCRNA H19 SPONGES MIR-106A-5P AND CONTRIBUTES TO ALDOSTERONE-INDUCED VASCULAR CALCIFICATION VIA ACTIVATING THE RUNX2-DEPENDENT PATHWAY. BACKGROUND: EXCESS ALDOSTERONE IS IMPLICATED IN VASCULAR CALCIFICATION (VC), BUT THE MECHANISM BY WHICH ALDOSTERONE-MR (MINERALOCORTICOID RECEPTOR) COMPLEX PROMOTES VC IS UNCLEAR. EMERGING EVIDENCE INDICATES THAT LONG-NONCODING RNA H19 (H19) PLAYS A CRITICAL ROLE IN VC. WE EXAMINED WHETHER ALDOSTERONE-INDUCED OSTEOGENIC DIFFERENTIATION OF VASCULAR SMOOTH MUSCLE CELLS (VSMCS) THROUGH H19 EPIGENETIC MODIFICATION OF RUNX2 (RUNT-RELATED TRANSCRIPTION FACTOR-2) IN A MR-DEPENDENT MANNER. METHODS: WE INDUCED IN VIVO RAT MODEL OF CHRONIC KIDNEY DISEASE USING A HIGH ADENINE AND PHOSPHATE DIET TO EXPLORE THE RELATIONSHIP AMONG ALDOSTERONE, MR, H19, AND VC. WE ALSO CULTURED HUMAN AORTIC VSMCS TO EXPLORE THE ROLES OF H19 IN ALDOSTERONE-MR COMPLEX-INDUCED OSTEOGENIC DIFFERENTIATION AND CALCIFICATION OF VSMCS. RESULTS: H19 AND RUNX2 WERE SIGNIFICANTLY INCREASED IN ALDOSTERONE-INDUCED VSMC OSTEOGENIC DIFFERENTIATION AND VC, BOTH IN VITRO AND IN VIVO, WHICH WERE SIGNIFICANTLY BLOCKED BY THE MR ANTAGONIST SPIRONOLACTONE. MECHANISTICALLY, OUR FINDINGS REVEAL THAT THE ALDOSTERONE-ACTIVATED MR BOUND TO H19 PROMOTER AND INCREASED ITS TRANSCRIPTIONAL ACTIVITY, AS DETERMINED BY CHROMATIN IMMUNOPRECIPITATION, ELECTROPHORETIC MOBILITY SHIFT ASSAY, AND LUCIFERASE REPORTER ASSAY. SILENCING H19 INCREASED MICRORNA-106A-5P (MIR-106A-5P) EXPRESSION, WHICH SUBSEQUENTLY INHIBITED ALDOSTERONE-INDUCED RUNX2 EXPRESSION AT THE POSTTRANSCRIPTIONAL LEVEL. IMPORTANTLY, WE OBSERVED A DIRECT INTERACTION BETWEEN H19 AND MIR-106A-5P, AND DOWNREGULATION OF MIR-106A-5P EFFICIENTLY REVERSED THE SUPPRESSION OF RUNX2 INDUCED BY H19 SILENCING. CONCLUSIONS: OUR STUDY CLARIFIES A NOVEL MECHANISM BY WHICH UPREGULATION OF H19 CONTRIBUTES TO ALDOSTERONE-MR COMPLEX-PROMOTED RUNX2-DEPENDENT VSMC OSTEOGENIC DIFFERENTIATION AND VC THROUGH SPONGING MIR-106A-5P. THESE FINDINGS HIGHLIGHT A POTENTIAL THERAPEUTIC TARGET FOR ALDOSTERONE-INDUCED VC. 2023 20 3946 33 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