1 1807 134 EFFECTIVE MODULATION OF INFLAMMATION AND OXIDATIVE STRESS FOR ENHANCED REGENERATION OF INTERVERTEBRAL DISCS USING 3D POROUS HYBRID PROTEIN NANOSCAFFOLD. DEGENERATION OF FIBROCARTILAGINOUS TISSUES IS OFTEN ASSOCIATED WITH COMPLEX PRO-INFLAMMATORY FACTORS. THESE INCLUDE REACTIVE OXYGEN SPECIES (ROS), CELL-FREE NUCLEIC ACIDS (CF-NAS), AND EPIGENETIC CHANGES IN IMMUNE CELLS. TO EFFECTIVELY CONTROL THIS COMPLEX INFLAMMATORY SIGNALING, IT DEVELOPED AN ALL-IN-ONE NANOSCAFFOLD-BASED 3D POROUS HYBRID PROTEIN (3D-PHP) SELF-THERAPEUTIC STRATEGY FOR TREATING INTERVERTEBRAL DISC (IVD) DEGENERATION. THE 3D-PHP NANOSCAFFOLD IS SYNTHESIZED BY INTRODUCING A NOVEL NANOMATERIAL-TEMPLATED PROTEIN ASSEMBLY (NTPA) STRATEGY. 3D-PHP NANOSCAFFOLDS THAT AVOID COVALENT MODIFICATION OF PROTEINS DEMONSTRATE INFLAMMATORY STIMULI-RESPONSIVE DRUG RELEASE, DISC-MIMETIC STIFFNESS, AND EXCELLENT BIODEGRADABILITY. ENZYME-LIKE 2D NANOSHEETS INCORPORATED INTO NANOSCAFFOLDS FURTHER ENABLED ROBUST SCAVENGING OF ROS AND CF-NAS, REDUCING INFLAMMATION AND ENHANCING THE SURVIVAL OF DISC CELLS UNDER INFLAMMATORY STRESS IN VITRO. IMPLANTATION OF 3D-PHP NANOSCAFFOLDS LOADED WITH BROMODOMAIN EXTRATERMINAL INHIBITOR (BETI) INTO A RAT NUCLEOTOMY DISC INJURY MODEL EFFECTIVELY SUPPRESSED INFLAMMATION IN VIVO, THUS PROMOTING RESTORATION OF THE EXTRACELLULAR MATRIX (ECM). THE RESULTING REGENERATION OF DISC TISSUE FACILITATED LONG-TERM PAIN REDUCTION. THEREFORE, SELF-THERAPEUTIC AND EPIGENETIC MODULATOR-ENCAPSULATED HYBRID PROTEIN NANOSCAFFOLD SHOWS GREAT PROMISE AS A NOVEL APPROACH TO RESTORE DYSREGULATED INFLAMMATORY SIGNALING AND TREAT DEGENERATIVE FIBROCARTILAGINOUS DISEASES, INCLUDING DISC INJURIES, PROVIDING HOPE AND RELIEF TO PATIENTS WORLDWIDE. 2023 2 3485 19 IDENTIFICATION OF CIRCULAR RNAS EXPRESSION PATTERN IN CAPRINE FETAL FIBROBLAST CELLS EXPOSED TO A CHRONIC NON-CYTOTOXIC DOSE OF GRAPHENE OXIDE-SILVER NANOPARTICLE NANOCOMPOSITES. THE WIDESPREAD USE OF GRAPHENE OXIDE-SILVER NANOPARTICLE NANOCOMPOSITES (GO-AGNPS) IN BIOMEDICAL SCIENCES IS INCREASING THE CHANCES OF HUMAN AND ANIMAL EXPOSURE TO ITS CHRONIC NON-TOXIC DOSES. EXPOSURE TO AGNPS-RELATED NANOMATERIALS MAY RESULT IN THE NEGATIVE EFFECT ON THE DAM, FETUS AND OFFSPRING. HOWEVER, THERE ARE ONLY LITTLE AVAILABLE INFORMATION FOR PROFOUND UNDERSTANDING OF THE EPIGENETIC ALTERATION IN THE CELLS AND ANIMALS CAUSED BY LOW-DOSE CHRONIC EXPOSURE OF GO-AGNPS. THE PRESENT STUDY INVESTIGATED THE EFFECT OF 0.5 MUG/ML GO-AGNPS FOR 10 WEEKS ON THE DIFFERENTIAL EXPRESSION OF CIRCULAR RNAS (CIRCRNAS) IN CAPRINE FETAL FIBROBLAST CELLS (CFFCS), AND THIS DOSE OF GO-AGNPS DID NOT AFFECT CELL VIABILITY AND ROS LEVEL. WE PREDICTED THE FUNCTIONS OF THOSE DIFFERENTIALLY EXPRESSED (DE) CIRCRNAS IN CFFCS BY BIOINFORMATICS ANALYSIS. FURTHERMORE, WE VALIDATED THE EXPRESSION OF TEN DE CIRCRNAS USING QUANTITATIVE REAL-TIME REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION (QRT-PCR) TO ENSURE THE RELIABILITY OF THE SEQUENCING DATA. OUR RESULTS SHOWED THAT THE DE CIRCRNAS MAY POTENTIALLY REGULATE THE GO-AGNPS-INDUCING EPIGENETIC TOXICITY THROUGH A REGULATORY NETWORK CONSISTED OF CIRCRNAS, MIRNAS AND MESSENGER RNAS (MRNAS). THEREFORE, THE EPIGENETICS TOXICITY IS ESSENTIAL TO ASSESS THE BIOSAFETY LEVEL OF GO-AGNPS. 2023 3 2284 14 EPIGENETIC REGULATION IN INTERVERTEBRAL DISC DEGENERATION. INTERVERTEBRAL DISC (IVD) DEGENERATION IS THE LEADING CAUSE OF LOW BACK PAIN, WHICH HAS A STRIKING IMPACT ON NUMEROUS PATIENTS. THEREFORE, COMPREHENSIVELY ILLUMINATING THE REGULATORY MECHANISMS OF IVD DEGENERATION IS OF GREAT SIGNIFICANCE. HERE, WE DISCUSS THE LATEST ADVANCES IN UNDERSTANDING THE MAIN EPIGENETIC MECHANISMS REGULATING IVD DEGENERATION. 2022 4 693 28 BREAKING BOUNDARIES: PAN BETI DISRUPT 3D CHROMATIN STRUCTURE, BD2-SELECTIVE BETI ARE STRICTLY EPIGENETIC TRANSCRIPTIONAL REGULATORS. BACKGROUND: BROMODOMAIN AND EXTRATERMINAL PROTEINS (BETS) ARE MORE THAN JUST EPIGENETIC REGULATORS OF TRANSCRIPTION. HERE WE HIGHLIGHT A NEW ROLE FOR THE BET PROTEIN BRD4 IN THE MAINTENANCE OF HIGHER ORDER CHROMATIN STRUCTURE AT TOPOLOGICALLY ASSOCIATING DOMAIN BOUNDARIES (TADBS). BD2-SELECTIVE AND PAN (NON-SELECTIVE) BET INHIBITORS (BETI) DIFFERENTIALLY SUPPORT CHROMATIN STRUCTURE, SELECTIVELY AFFECTING TRANSCRIPTION AND CELL VIABILITY. METHODS: USING RNA-SEQ AND BRD4 CHIP-SEQ, THE DIFFERENTIAL EFFECT OF BETI TREATMENT ON THE TRANSCRIPTOME AND BRD4 CHROMATIN OCCUPANCY OF HUMAN AORTIC ENDOTHELIAL CELLS FROM DIABETIC PATIENTS (DHAECS) STIMULATED WITH TNFALPHA WAS EVALUATED. CHROMATIN DECONDENSATION AND DNA FRAGMENTATION WAS ASSESSED BY IMMUNOFLUORESCENCE IMAGING AND QUANTIFICATION. KEY DHAEC FINDINGS WERE VERIFIED IN PROLIFERATING MONOCYTE-LIKE THP-1 CELLS USING REAL TIME-PCR, BRD4 CO-IMMUNOPRECIPITATION STUDIES, WESTERN BLOTS, PROLIFERATION AND APOPTOSIS ASSAYS. FINDINGS: WE DISCOVERED THAT 1) BRD4 CO-LOCALIZES WITH YING-YANG 1 (YY1) AT TADBS, CRITICAL CHROMATIN STRUCTURE COMPLEXES PROXIMAL TO MANY DNA REPAIR GENES. 2) BD2-SELECTIVE BETI ENRICH BRD4/YY1 ASSOCIATIONS, WHILE PAN-BETI DO NOT. 3) FAILURE TO SUPPORT CHROMATIN STRUCTURES THROUGH BRD4/YY1 ENRICHMENT INHIBITS DNA REPAIR GENE TRANSCRIPTION, WHICH INDUCES DNA DAMAGE RESPONSES, AND CAUSES WIDESPREAD CHROMATIN DECONDENSATION, DNA FRAGMENTATION, AND APOPTOSIS. 4) BD2-SELECTIVE BETI MAINTAIN HIGH ORDER CHROMATIN STRUCTURE AND CELL VIABILITY, WHILE REDUCING DELETERIOUS PRO-INFLAMMATORY TRANSCRIPTION. INTERPRETATION: BRD4 PLAYS A PREVIOUSLY UNRECOGNIZED ROLE AT TADBS. BETI DIFFERENTIALLY IMPACT TADB STABILITY. OUR RESULTS PROVIDE TRANSLATIONAL INSIGHT FOR THE DEVELOPMENT OF BETI AS THERAPEUTICS FOR A RANGE OF DISEASES INCLUDING CVD, CHRONIC KIDNEY DISEASE, CANCER, AND COVID-19. 2022 5 4033 29 M6A HYPOMETHYLATION OF DNMT3B REGULATED BY ALKBH5 PROMOTES INTERVERTEBRAL DISC DEGENERATION VIA E4F1 DEFICIENCY. BACKGROUND: THE INTERVERTEBRAL DISC (IVD) DEGENERATION IS THE LEADING CAUSE OF LOW BACK PAIN, WHICH ACCOUNTS FOR A MAIN CAUSE OF DISABILITY. N6-METHYLADENOSINE (M6A) IS THE MOST ABUNDANT INTERNAL MODIFICATION IN EUKARYOTIC MESSENGER RNAS AND IS INVOLVED IN VARIOUS DISEASES AND CELLULAR PROCESSES BY MODULATING MRNA FATE. HOWEVER, THE CRITICAL ROLE OF M6A REGULATION IN IVD DEGENERATION REMAINS UNCLEAR. NUCLEUS PULPOSUS CELL (NPC) SENESCENCE IS CRITICAL FOR THE PROGRESSION OF IVD DEGENERATION. HERE, WE UNCOVERED THE ROLE AND EXPLORED THE REGULATORY MECHANISM OF M6A IN NPC SENESCENCE DURING IVD DEGENERATION. METHODS: IDENTIFICATION OF NPC SENESCENCE DURING IVD DEGENERATION WAS BASED ON THE ANALYSIS OF TISSUE SAMPLES AND THE CELLULAR MODEL. ALKBH5 UPREGULATION INDUCING CELLULAR SENESCENCE WAS CONFIRMED BY FUNCTIONAL EXPERIMENTS IN VIVO AND IN VITRO. CHIP-QPCR AND DNA-PULLDOWN WERE USED TO REVEAL INCREASED ALKBH5 WAS REGULATED BY KDM4A-MEDIATED H3K9ME3. FURTHERMORE, ME-RIP-SEQ WAS PERFORMED TO IDENTIFY M6A HYPOMETHYLATION OF DNMT3B TRANSCRIPTS IN SENESCENT NPCS. STABILITY ANALYSIS SHOWED THAT DNMT3B EXPRESSION WAS ENHANCED FOR LESS YTHDF2 RECOGNITION AND INCREASED DNMT3B PROMOTED NPC SENESCENCE AND IVD DEGENERATION VIA E4F1 METHYLATION BY IN VIVO AND IN VITRO ANALYSES. RESULTS: EXPRESSION OF ALKBH5 IS ENHANCED DURING IVD DEGENERATION AND NPC SENESCENCE, DUE TO DECREASED KDM4A-MEDIATED H3K9ME3 MODIFICATION. FUNCTIONALLY, ALKBH5 CAUSES NPC SENESCENCE BY DEMETHYLATING DNMT3B TRANSCRIPTS AND IN TURN PROMOTING ITS EXPRESSION VIA LESS YTHDF2 RECOGNITION AND FOLLOWING DEGRADATION DUE TO TRANSCRIPT HYPOMETHYLATION IN VITRO AND IN VIVO. INCREASED DNMT3B PROMOTES THE DEVELOPMENT OF IVD DEGENERATION AND NPC SENESCENCE, MECHANISTICALLY BY METHYLATING CPG ISLANDS OF E4F1 AT THE PROMOTER REGION AND THUS RESTRAINING ITS TRANSCRIPTION AND EXPRESSION. CONCLUSIONS: COLLECTIVELY, OUR FINDINGS REVEAL AN EPIGENETIC INTERPLAY MECHANISM IN NPC SENESCENCE AND IVD DEGENERATION, PRESENTING A CRITICAL PRO-SENESCENCE ROLE OF ALKBH5 AND M6A HYPOMETHYLATION, HIGHLIGHTING THE THERAPEUTIC POTENTIAL OF TARGETING THE M6A/DNMT3B/E4F1 AXIS FOR TREATING IVD DEGENERATION. 2022 6 699 27 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 7 1826 31 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 8 4366 20 MIRNA-23A/CXCR4 REGULATES NEUROPATHIC PAIN VIA DIRECTLY TARGETING TXNIP/NLRP3 INFLAMMASOME AXIS. BACKGROUND: CHEMOKINE CXC RECEPTOR 4 (CXCR4) IN SPINAL GLIAL CELLS HAS BEEN IMPLICATED IN NEUROPATHIC PAIN. HOWEVER, THE REGULATORY CASCADES OF CXCR4 IN NEUROPATHIC PAIN REMAIN ELUSIVE. HERE, WE INVESTIGATED THE FUNCTIONAL REGULATORY ROLE OF MIRNAS IN THE PAIN PROCESS AND ITS INTERPLAY WITH CXCR4 AND ITS DOWNSTREAM SIGNALING. METHODS: MIRNAS AND CXCR4 AND ITS DOWNSTREAM SIGNALING MOLECULES WERE MEASURED IN THE SPINAL CORDS OF MICE WITH SCIATIC NERVE INJURY VIA PARTIAL SCIATIC NERVE LIGATION (PSNL). IMMUNOBLOTTING, IMMUNOFLUORESCENCE, IMMUNOPRECIPITATION, AND MAMMAL TWO-HYBRID AND BEHAVIORAL TESTS WERE USED TO EXPLORE THE DOWNSTREAM CXCR4-DEPENDENT SIGNALING PATHWAY. RESULTS: CXCR4 EXPRESSION INCREASED IN SPINAL GLIAL CELLS OF MICE WITH PSNL-INDUCED NEUROPATHIC PAIN. BLOCKING CXCR4 ALLEVIATED THE PAIN BEHAVIOR; CONTRARILY, OVEREXPRESSING CXCR4 INDUCED PAIN HYPERSENSITIVITY. MICRORNA-23A-3P (MIR-23A) DIRECTLY BOUNDS TO 3' UTR OF CXCR4 MRNA. PSNL-INDUCED NEUROPATHIC PAIN SIGNIFICANTLY REDUCED MRNA EXPRESSION OF MIR-23A. OVEREXPRESSION OF MIR-23A BY INTRATHECAL INJECTION OF MIR-23A MIMICS OR LENTIVIRUS REDUCED SPINAL CXCR4 AND PREVENTED PSNL-INDUCED NEUROPATHIC PAIN. IN CONTRAST, KNOCKDOWN OF MIR-23A BY INTRATHECAL INJECTION OF MIR-23A INHIBITOR OR LENTIVIRUS INDUCED PAIN-LIKE BEHAVIOR, WHICH WAS REDUCED BY CXCR4 INHIBITION. ADDITIONALLY, MIR-23A KNOCKDOWN OR CXCR4 OVEREXPRESSION IN NAIVE MICE COULD INCREASE THE THIOREDOXIN-INTERACTING PROTEIN (TXNIP), WHICH WAS ASSOCIATED WITH INDUCTION OF NOD-LIKE RECEPTOR PROTEIN 3 (NLRP3) INFLAMMASOME. INDEED, CXCR4 AND TXNIP WERE CO-EXPRESSED. THE MAMMAL TWO-HYBRID ASSAY REVEALED THE DIRECT INTERACTION BETWEEN CXCR4 AND TXNIP, WHICH WAS INCREASED IN THE SPINAL CORD OF PSNL MICE. IN PARTICULAR, INHIBITION OF TXNIP REVERSED PAIN BEHAVIOR ELICITED BY PSNL, MIR-23A KNOCKDOWN, OR CXCR4 OVEREXPRESSION. MOREOVER, MIR-23A OVEREXPRESSION OR CXCR4 KNOCKDOWN INHIBITED THE INCREASE OF TXNIP AND NLRP3 INFLAMMASOME IN PSNL MICE. CONCLUSIONS: MIR-23A, BY DIRECTLY TARGETING CXCR4, REGULATES NEUROPATHIC PAIN VIA TXNIP/NLRP3 INFLAMMASOME AXIS IN SPINAL GLIAL CELLS. EPIGENETIC INTERVENTIONS AGAINST MIR-23A, CXCR4, OR TXNIP MAY POTENTIALLY SERVE AS NOVEL THERAPEUTIC AVENUES IN TREATING PERIPHERAL NERVE INJURY-INDUCED NOCICEPTIVE HYPERSENSITIVITY. 2018 9 692 28 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 10 6580 21 TREPONEMA DENTICOLA INCREASES MMP-2 EXPRESSION AND ACTIVATION IN THE PERIODONTIUM VIA REVERSIBLE DNA AND HISTONE MODIFICATIONS. HOST-DERIVED MATRIX METALLOPROTEINASES (MMPS) AND BACTERIAL PROTEASES MEDIATE DESTRUCTION OF EXTRACELLULAR MATRICES AND SUPPORTING ALVEOLAR BONE IN PERIODONTITIS. THE TREPONEMA DENTICOLA DENTILISIN PROTEASE INDUCES MMP-2 EXPRESSION AND ACTIVATION IN PERIODONTAL LIGAMENT (PDL) CELLS, AND DENTILISIN-MEDIATED ACTIVATION OF PRO-MMP-2 IS REQUIRED FOR CELLULAR FIBRONECTIN DEGRADATION. HERE, WE REPORT THAT T. DENTICOLA REGULATES MMP-2 EXPRESSION THROUGH EPIGENETIC MODIFICATIONS IN THE PERIODONTIUM. PDL CELLS WERE TREATED WITH EPIGENETIC ENZYME INHIBITORS BEFORE OR AFTER T. DENTICOLA CHALLENGE. FIBRONECTIN FRAGMENTATION, MMP-2 EXPRESSION, AND ACTIVATION WERE ASSESSED BY IMMUNOBLOT, ZYMOGRAPHY, AND QRT-PCR, RESPECTIVELY. CHROMATIN MODIFICATION ENZYME EXPRESSION IN T. DENTICOLA-CHALLENGED PDL CELLS AND PERIODONTAL TISSUES WERE EVALUATED USING GENE ARRAYS. SEVERAL CLASSES OF EPIGENETIC ENZYMES SHOWED SIGNIFICANT ALTERATIONS IN TRANSCRIPTION IN DISEASED TISSUE AND T. DENTICOLA-CHALLENGED PDL CELLS. T. DENTICOLA-MEDIATED MMP-2 EXPRESSION AND ACTIVATION WERE SIGNIFICANTLY REDUCED IN PDL CELLS TREATED WITH INHIBITORS OF AURORA KINASES AND HISTONE DEACETYLASES. IN CONTRAST, DNA METHYLTRANSFERASE INHIBITORS HAD LITTLE EFFECT, AND INHIBITORS OF HISTONE ACETYLTRANSFERASES, METHYLTRANSFERASES, AND DEMETHYLASES EXACERBATED T. DENTICOLA-MEDIATED MMP-2 EXPRESSION AND ACTIVATION. CHRONIC EPIGENETIC CHANGES IN PERIODONTAL TISSUES MEDIATED BY T. DENTICOLA OR OTHER ORAL MICROBES MAY CONTRIBUTE TO THE LIMITED SUCCESS OF CONVENTIONAL TREATMENT OF CHRONIC PERIODONTITIS AND MAY BE AMENABLE TO THERAPEUTIC REVERSAL. 2018 11 4529 20 MULTIGENERATIONAL GRAPHENE OXIDE INTOXICATION RESULTS IN REPRODUCTION DISORDERS AT THE MOLECULAR LEVEL OF VITELLOGENIN PROTEIN EXPRESSION IN ACHETA DOMESTICUS. THE ANTHROPOGENIC ACTIVITIES MAY LEAD TO ACCUMULATION OF GRAPHENE OXIDE (GO) POLLUTION IN THE ENVIRONMENT. ORGANISMS EXPOSED TO CHRONIC OR MULTIGENERATIONAL GO INTOXICATION CAN PRESENT REPRODUCTION DEPLETION. VITELLOGENIN (VG) HAS BEEN USED AS A PARAMETER FOR EVALUATING FEMALE FERTILITY DUE TO ITS IMPORTANCE IN EMBRYO NUTRITION. IN THIS STUDY, WE USED A PROMISING MODEL ORGANISM, ACHETA DOMESTICUS, WHICH WAS INTOXICATED WITH GO IN FOOD FOR THREE GENERATIONS. THE AIM OF THE STUDY WAS TO INVESTIGATE THE PROCESS OF VG SYNTHESIS IN CRICKETS DEPENDING ON THE EXPOSURE TIME, GO CONCENTRATION, AND AGE OF THE FEMALES. THE RESULTS REVEALED THAT CHRONIC GO INTOXICATION HAD ADVERSE EFFECTS ON THE VG EXPRESSION PATTERN. THE 1ST GENERATION OF INSECTS SHOWING LOW VG EXPRESSION WAS MOST AFFECTED. THE 2ND GENERATION OF A. DOMESTICUS PRESENTED A HIGH VG EXPRESSION. THE LAST INVESTIGATED GENERATION SEEMED TO COPE WITH STRESS CAUSED BY GO, AND THE VG EXPRESSION WAS BALANCED. WE SUGGEST THAT THE EPIGENETIC MECHANISMS MAY PLAY A ROLE IN THE INFORMATION TRANSFER TO THE NEXT GENERATIONS ON HOW TO REACT TO THE RISK FACTOR AND KEEP REPRODUCTION AT A HIGH RATE. WE SUSPECT THAT CHRONIC GO INTOXICATION CAN DISTURB THE REGULAR FORMATION OF THE VG QUATERNARY STRUCTURE, RESULTING IN CONSEQUENCES FOR DEVELOPING AN EMBRYO. 2021 12 2784 26 EZH2 PROMOTES EXTRACELLULAR MATRIX DEGRADATION VIA NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN PULPITIS. PULPITIS IS A COMPLICATED CHRONIC INFLAMMATORY PROCESS WHICH CAN BE IN A DYNAMIC BALANCE BETWEEN DAMAGE AND REPAIR. THE EXTRACELLULAR MATRIX PLAYS AN IMPORTANT REGULATORY ROLE IN WOUND HEALING AND TISSUE REPAIR. THE AIM OF THIS STUDY WAS TO EXPLORE THE ROLE OF THE EPIGENETIC MARK, ENHANCER OF ZESTE HOMOLOG 2 (EZH2) ON THE DEGRADATION OF EXTRACELLULAR MATRIX DURING PULPITIS. QUANTITATIVE POLYMERASE CHAIN REACTION WAS USED TO ASSESS THE EXPRESSION OF MATRIX METALLOPROTEINASES (MMPS) AND TYPE I COLLAGEN IN HUMAN DENTAL PULP CELLS (HDPCS) UPON EZH2 AND EI1 (EZH2 INHIBITOR) STIMULATION. THE MECHANISM OF EZH2 AFFECTING EXTRACELLULAR MATRIX WAS EXPLORED THROUGH QUANTITATIVE POLYMERASE CHAIN REACTION AND WESTERN BLOT. A RAT MODEL OF DENTAL PULP INFLAMMATION WAS ESTABLISHED, AND THE EXPRESSION OF TYPE I COLLAGEN IN DENTAL PULP UNDER EZH2 STIMULATION WAS DETECTED BY IMMUNOHISTOCHEMICAL STAINING. EZH2 UPREGULATED THE EXPRESSION OF MMP-1, MMP-3, MMP-8, AND MMP-10 AND DECREASED THE PRODUCTION OF TYPE I COLLAGEN IN HDPCS, WHILE EI1 HAD THE OPPOSITE EFFECT. EZH2 ACTIVATED THE NUCLEAR FACTOR-KAPPA B (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN HDPCS, THE INHIBITION OF WHICH REVERSED THE INDUCTION OF MMPS AND THE SUPPRESSION OF TYPE I COLLAGEN. EZH2 CAN DOWNREGULATE THE TYPE I COLLAGEN LEVELS IN AN EXPERIMENTAL MODEL OF DENTAL PULPITIS IN RATS. EZH2 PROMOTES EXTRACELLULAR MATRIX DEGRADATION VIA NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN PULPITIS. EZH2 CAN DECREASE THE TYPE I COLLAGEN LEVELS IN VIVO AND IN VITRO. 2021 13 4312 16 MICRORNAS AS A SUITABLE BIOMARKER TO DETECT THE EFFECTS OF LONG-TERM EXPOSURES TO NANOMATERIALS. STUDIES ON TIO(2)NP AND MWCNT. THE PRESENCE OF NANOMATERIALS (NMS) IN THE ENVIRONMENT MAY REPRESENT A SERIOUS RISK TO HUMAN HEALTH, ESPECIALLY IN A SCENARIO OF CHRONIC EXPOSURE. TO EVALUATE THE POTENTIAL RELATIONSHIP BETWEEN NM-INDUCED EPIGENETIC ALTERATIONS AND CARCINOGENESIS, THE PRESENT STUDY ANALYZED A PANEL OF 33 MIRNAS RELATED TO THE CELL TRANSFORMATION PROCESS IN BEAS-2B CELLS TRANSFORMED BY TIO(2)NP AND LONG-TERM MWCNT EXPOSURE. OUR BATTERY REVEALED A LARGE IMPACT ON MIRNA EXPRESSION PROFILING IN CELLS EXPOSED TO BOTH NMS. FROM THIS ANALYSIS, A SMALL SET OF FIVE MIRNAS (MIR-23A, MIR-25, MIR-96, MIR-210, AND MIR-502) WERE IDENTIFIED AS INFORMATIVE BIOMARKERS OF THE TRANSFORMING EFFECTS INDUCED BY NM EXPOSURES. THE USEFULNESS OF THIS REDUCED MIRNA BATTERY WAS FURTHER VALIDATED IN OTHER PREVIOUSLY GENERATED TRANSFORMED CELL SYSTEMS BY LONG-TERM EXPOSURE TO OTHER NMS (CONP, ZNONP, MSINP, AND CEO(2)NP). INTERESTINGLY, THE FIVE SELECTED MIRNAS WERE CONSISTENTLY OVEREXPRESSED IN ALL CELL LINES AND NMS TESTED. THESE RESULTS CONFIRM THE SUITABILITY OF THE PROPOSED SET OF MRNAS TO IDENTIFY THE POTENTIAL TRANSFORMING ABILITY OF NMS. PARTICULAR ATTENTION SHOULD BE PAID TO THE EPIGENOME AND ESPECIALLY TO MIRNAS FOR HAZARD ASSESSMENT OF NMS, AS WELLS AS FOR THE STUDY OF THE UNDERLYING MECHANISMS OF ACTION. 2021 14 1764 27 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 15 3614 22 IN VITRO CELL TRANSFORMATION ASSAYS: A VALUABLE APPROACH FOR CARCINOGENIC POTENTIALITY ASSESSMENT OF NANOMATERIALS. THIS REVIEW EXPLORES THE APPLICATION OF IN VITRO CELL TRANSFORMATION ASSAYS (CTAS) AS A SCREENING PLATFORM TO ASSESS THE CARCINOGENIC POTENTIAL OF NANOMATERIALS (NMS) RESULTING FROM CONTINUOUSLY GROWING INDUSTRIAL PRODUCTION AND USE. THE WIDESPREAD APPLICATION OF NMS IN VARIOUS FIELDS HAS RAISED CONCERNS ABOUT THEIR POTENTIAL ADVERSE EFFECTS, NECESSITATING SAFETY EVALUATIONS, PARTICULARLY IN LONG-TERM CONTINUOUS EXPOSURE SCENARIOS. CTAS PRESENT A REALISTIC SCREENING PLATFORM FOR KNOWN AND EMERGING NMS BY EXAMINING THEIR RESEMBLANCE TO THE HALLMARK OF MALIGNANCY, INCLUDING HIGH PROLIFERATION RATES, LOSS OF CONTACT INHIBITION, THE GAIN OF ANCHORAGE-INDEPENDENT GROWTH, CELLULAR INVASION, DYSREGULATION OF THE CELL CYCLE, APOPTOSIS RESISTANCE, AND ABILITY TO FORM TUMORS IN EXPERIMENTAL ANIMALS. THROUGH THE DELIBERATE TRANSFORMATION OF CELLS VIA CHRONIC NM EXPOSURE, RESEARCHERS CAN INVESTIGATE THE TUMORIGENIC PROPERTIES OF NMS AND THE UNDERLYING MECHANISMS OF CANCER DEVELOPMENT. THIS ARTICLE EXAMINES NM-INDUCED CELL TRANSFORMATION STUDIES, FOCUSING ON IDENTIFYING EXISTING KNOWLEDGE GAPS. SPECIFICALLY, IT EXPLORES THE PHYSICOCHEMICAL PROPERTIES OF NMS, EXPERIMENTAL MODELS, ASSAYS, DOSE AND TIME REQUIREMENTS FOR CELL TRANSFORMATION, AND THE UNDERLYING MECHANISMS OF MALIGNANCY. OUR REVIEW AIMS TO ADVANCE UNDERSTANDING IN THIS FIELD AND IDENTIFY AREAS FOR FURTHER INVESTIGATION. 2023 16 589 23 BET BROMODOMAIN INHIBITORS SUPPRESS INFLAMMATORY ACTIVATION OF GINGIVAL FIBROBLASTS AND EPITHELIAL CELLS FROM PERIODONTITIS PATIENTS. BET BROMODOMAIN PROTEINS ARE IMPORTANT EPIGENETIC REGULATORS OF GENE EXPRESSION THAT BIND ACETYLATED HISTONE TAILS AND REGULATE THE FORMATION OF ACETYLATION-DEPENDENT CHROMATIN COMPLEXES. BET INHIBITORS SUPPRESS INFLAMMATORY RESPONSES IN MULTIPLE CELL TYPES AND ANIMAL MODELS, AND PROTECT AGAINST BONE LOSS IN EXPERIMENTAL PERIODONTITIS IN MICE. HERE, WE ANALYZED THE ROLE OF BET PROTEINS IN INFLAMMATORY ACTIVATION OF GINGIVAL FIBROBLASTS (GFS) AND GINGIVAL EPITHELIAL CELLS (GECS). WE SHOW THAT THE BET INHIBITORS I-BET151 AND JQ1 SIGNIFICANTLY REDUCED EXPRESSION AND/OR PRODUCTION OF DISTINCT, BUT OVERLAPPING, PROFILES OF CYTOKINE-INDUCIBLE MEDIATORS OF INFLAMMATION AND BONE RESORPTION IN GFS FROM HEALTHY DONORS (IL6, IL8, IL1B, CCL2, CCL5, COX2, AND MMP3) AND THE GEC LINE TIGK (IL6, IL8, IL1B, CXCL10, MMP9) WITHOUT AFFECTING CELL VIABILITY. ACTIVATION OF MITOGEN-ACTIVATED PROTEIN KINASE AND NUCLEAR FACTOR-KAPPAB PATHWAYS WAS UNAFFECTED BY I-BET151, AS WAS THE HISTONE ACETYLATION STATUS, AND NEW PROTEIN SYNTHESIS WAS NOT REQUIRED FOR THE ANTI-INFLAMMATORY EFFECTS OF BET INHIBITION. I-BET151 AND JQ1 ALSO SUPPRESSED EXPRESSION OF INFLAMMATORY CYTOKINES, CHEMOKINES, AND OSTEOCLASTOGENIC MEDIATORS IN GFS AND TIGKS INFECTED WITH THE KEY PERIODONTAL PATHOGEN PORPHYROMONAS GINGIVALIS. NOTABLY, P. GINGIVALIS INTERNALIZATION AND INTRACELLULAR SURVIVAL IN GFS AND TIGKS REMAINED UNAFFECTED BY BET INHIBITORS. FINALLY, INHIBITION OF BET PROTEINS SIGNIFICANTLY REDUCED P. GINGIVALIS-INDUCED INFLAMMATORY MEDIATOR EXPRESSION IN GECS AND GFS FROM PATIENTS WITH PERIODONTITIS. OUR RESULTS DEMONSTRATE THAT BET INHIBITORS MAY BLOCK THE EXCESSIVE INFLAMMATORY MEDIATOR PRODUCTION BY RESIDENT CELLS OF THE GINGIVAL TISSUE AND IDENTIFY THE BET FAMILY OF EPIGENETIC READER PROTEINS AS A POTENTIAL THERAPEUTIC TARGET IN THE TREATMENT OF PERIODONTAL DISEASE. 2019 17 6687 21 VALIDATION OF THE EPIGENETIC READER BROMODOMAIN-CONTAINING PROTEIN 4 (BRD4) AS A THERAPEUTIC TARGET FOR TREATMENT OF AIRWAY REMODELING. STRUCTURAL REMODELING IS CENTRAL TO THE INITIATION AND PROGRESSION OF MANY CHRONIC LUNG DISEASES, REPRESENTING AN IMPORTANT UNMET NEED. WE EXAMINE THE EVIDENCE SUPPORTING BROMODOMAIN-CONTAINING PROTEIN 4 (BRD4) AS A VALIDATED BIOLOGICAL TARGET FOR TREATMENT OF AIRWAY REMODELING. IN EPITHELIAL CELLS AND FIBROBLASTS, BRD4 SERVES AS A SCAFFOLD FOR CHROMATIN REMODELING COMPLEXES IN ACTIVE SUPER-ENHANCERS. IN RESPONSE TO INFLAMMATORY STIMULI, BRD4 IS REPOSITIONED TO INNATE AND MESENCHYMAL GENES ACTIVATING THEIR PRODUCTION. PROOF-OF-CONCEPT STUDIES SHOW PROMISING BENEFIT OF SELECTIVE BRD4 INHIBITORS IN DISRUPTING EPITHELIAL MESENCHYMAL TRANSITION AND MYOFIBROBLAST TRANSITION IN DIVERSE MODELS OF LUNG INJURY. RECENT IDENTIFICATION OF BIOMARKERS OF BRD4 PROVIDES A BASIS FOR FURTHER DRUG DEVELOPMENT FOR APPLICATION IN VIRAL-INDUCED AIRWAY INFLAMMATION, COPD AND INTERSTITIAL LUNG DISEASES. 2020 18 6023 25 THE BET PROTAC INHIBITOR DBET6 PROTECTS AGAINST RETINAL DEGENERATION AND INHIBITS THE CGAS-STING IN RESPONSE TO LIGHT DAMAGE. BACKGROUND: CHRONIC INFLAMMATION SIGNIFICANTLY CONTRIBUTES TO PHOTORECEPTOR DEATH IN BLINDING RETINAL DISEASES SUCH AS AGE-RELATED MACULAR DEGENERATION (AMD) AND RETINITIS PIGMENTOSA (RP). BROMODOMAIN AND EXTRATERMINAL DOMAIN (BET) PROTEINS ARE EPIGENETIC READERS THAT ACT AS KEY PROINFLAMMATORY FACTORS. WE RECENTLY FOUND THE FIRST-GENERATION BET INHIBITOR JQ1 ALLEVIATED SODIUM IODATE-INDUCED RETINAL DEGENERATION BY SUPPRESSING CGAS-STING INNATE IMMUNITY. HERE, WE INVESTIGATED THE EFFECTS AND MECHANISM OF DBET6, A PROTEOLYSIS?TARGETING CHIMERA (PROTAC) SMALL MOLECULE THAT SELECTIVELY DEGRADES BET BY THE UBIQUITIN?PROTEASOME SYSTEM, IN LIGHT-INDUCED RETINAL DEGENERATION. METHODS: MICE WERE EXPOSED TO BRIGHT LIGHT TO INDUCE RETINAL DEGENERATION, AND THE ACTIVATION OF CGAS-STING WAS DETERMINED BY RNA-SEQUENCING AND MOLECULAR BIOLOGY. RETINAL FUNCTION, MORPHOLOGY, PHOTORECEPTOR VIABILITY AND RETINAL INFLAMMATION WERE EXAMINED IN THE PRESENCE AND ABSENCE OF DBET6 TREATMENT. RESULTS: INTRAPERITONEAL INJECTION OF DBET6 LED TO THE RAPID DEGRADATION OF BET PROTEIN IN THE RETINA WITHOUT DETECTABLE TOXICITY. DBET6 IMPROVED RETINAL RESPONSIVENESS AND VISUAL ACUITY AFTER LIGHT DAMAGE (LD). DBET6 ALSO REPRESSED LD-INDUCED RETINAL MACROPHAGES/MICROGLIA ACTIVATION, MULLER CELL GLIOSIS, PHOTORECEPTOR DEATH AND RETINAL DEGENERATION. ANALYSIS OF SINGLE-CELL RNA-SEQUENCING RESULTS REVEALED CGAS-STING COMPONENTS WERE EXPRESSED IN RETINAL MICROGLIA. LD LED TO DRAMATIC ACTIVATION OF THE CGAS-STING PATHWAY, WHEREAS DBET6 SUPPRESSED LD-INDUCED STING EXPRESSION IN REACTIVE MACROPHAGES/MICROGLIA AND THE RELATED INFLAMMATORY RESPONSE. CONCLUSIONS: THIS STUDY INDICATES TARGETED DEGRADATION OF BET BY DBET6 EXERTS NEUROPROTECTIVE EFFECTS BY INHIBITING CGAS-STING IN REACTIVE RETINAL MACROPHAGES/MICROGLIA, AND IS EXPECTED TO BECOME A NEW STRATEGY FOR TREATMENT OF RETINAL DEGENERATION. 2023 19 5120 17 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 20 592 23 BET BROMODOMAIN PROTEINS REGULATE TRANSCRIPTIONAL REPROGRAMMING IN GENETIC DILATED CARDIOMYOPATHY. THE BROMODOMAIN AND EXTRATERMINAL (BET) FAMILY COMPRISES EPIGENETIC READER PROTEINS THAT ARE IMPORTANT REGULATORS OF INFLAMMATORY AND HYPERTROPHIC GENE EXPRESSION IN THE HEART. WE PREVIOUSLY IDENTIFIED THE ACTIVATION OF PROINFLAMMATORY GENE NETWORKS AS A KEY EARLY DRIVER OF DILATED CARDIOMYOPATHY (DCM) IN TRANSGENIC MICE EXPRESSING A MUTANT FORM OF PHOSPHOLAMBAN (PLNR9C) - A GENETIC CAUSE OF DCM IN HUMANS. WE HYPOTHESIZED THAT BETS COACTIVATE THIS INFLAMMATORY PROCESS, REPRESENTING A CRITICAL NODE IN THE PROGRESSION OF DCM. TO TEST THIS HYPOTHESIS, WE TREATED PLNR9C OR AGE-MATCHED WT MICE LONGITUDINALLY WITH THE SMALL MOLECULE BET BROMODOMAIN INHIBITOR JQ1 OR VEHICLE. BET INHIBITION ABROGATED ADVERSE CARDIAC REMODELING, REDUCED CARDIAC FIBROSIS, AND PROLONGED SURVIVAL IN PLNR9C MICE BY INHIBITING EXPRESSION OF PROINFLAMMATORY GENE NETWORKS AT ALL STAGES OF DISEASE. SPECIFICALLY, JQ1 HAD PROFOUND EFFECTS ON PROINFLAMMATORY GENE NETWORK EXPRESSION IN CARDIAC FIBROBLASTS, WHILE HAVING LITTLE EFFECT ON GENE EXPRESSION IN CARDIOMYOCYTES. CARDIAC FIBROBLAST PROLIFERATION WAS ALSO SUBSTANTIALLY REDUCED BY JQ1. MECHANISTICALLY, WE DEMONSTRATED THAT BRD4 SERVES AS A DIRECT AND ESSENTIAL REGULATOR OF NF-KAPPAB-MEDIATED PROINFLAMMATORY GENE EXPRESSION IN CARDIAC FIBROBLASTS. SUPPRESSING PROINFLAMMATORY GENE EXPRESSION VIA BET BROMODOMAIN INHIBITION COULD BE A NOVEL THERAPEUTIC STRATEGY FOR CHRONIC DCM IN HUMANS. 2020