1 589 132 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 2 3204 73 HDAC3 REGULATES GINGIVAL FIBROBLAST INFLAMMATORY RESPONSES IN PERIODONTITIS. HISTONE DEACETYLASES (HDACS) ARE IMPORTANT REGULATORS OF GENE EXPRESSION THAT ARE ABERRANTLY REGULATED IN SEVERAL INFLAMMATORY AND INFECTIOUS DISEASES. HDAC INHIBITORS (HDACI) SUPPRESS INFLAMMATORY ACTIVATION OF VARIOUS CELL TYPES THROUGH EPIGENETIC AND NON-EPIGENETIC MECHANISMS, AND AMELIORATE PATHOLOGY IN A MOUSE MODEL OF PERIODONTITIS. ACTIVATION OF GINGIVAL FIBROBLASTS (GFS) SIGNIFICANTLY CONTRIBUTES TO THE DEVELOPMENT OF PERIODONTITIS AND THE ANAEROBIC BACTERIUM PORPHYROMONAS GINGIVALIS PLAYS A KEY ROLE IN DRIVING CHRONIC INFLAMMATION. HERE, WE ANALYZED THE ROLE OF HDACS IN INFLAMMATORY RESPONSES OF GFS. PAN-HDACI SUBEROYLANILIDE HYDROXAMIC ACID (SAHA) AND/OR ITF2357 (GIVINOSTAT) SIGNIFICANTLY REDUCED TNFALPHA- AND P. GINGIVALIS-INDUCIBLE EXPRESSION AND/OR PRODUCTION OF A CLUSTER OF INFLAMMATORY MEDIATORS IN HEALTHY DONOR GFS (IL1B, CCL2, CCL5, CXCL10, COX2, AND MMP3) WITHOUT AFFECTING CELL VIABILITY. SELECTIVE INHIBITION OF HDAC3/6, BUT NOT SPECIFIC HDAC1, HDAC6, OR HDAC8 INHIBITION, REPRODUCED THE SUPPRESSIVE EFFECTS OF PAN-HDACI ON THE INFLAMMATORY GENE EXPRESSION PROFILE INDUCED BY TNFALPHA AND P. GINGIVALIS, SUGGESTING A CRITICAL ROLE FOR HDAC3 IN GF INFLAMMATORY ACTIVATION. CONSISTENTLY, SILENCING OF HDAC3 EXPRESSION WITH SIRNA LARGELY RECAPITULATED THE EFFECTS OF HDAC3/6I ON MRNA LEVELS OF INFLAMMATORY MEDIATORS IN P. GINGIVALIS-INFECTED GFS. IN CONTRAST, P. GINGIVALIS INTERNALIZATION AND INTRACELLULAR SURVIVAL IN GFS REMAINED UNAFFECTED BY HDACI. ACTIVATION OF MITOGEN-ACTIVATED PROTEIN KINASES AND NFKAPPAB SIGNALING WAS UNAFFECTED BY GLOBAL OR HDAC3/6-SELECTIVE HDACI, AND NEW PROTEIN SYNTHESIS WAS NOT REQUIRED FOR GENE SUPPRESSION BY HDACI. FINALLY, PAN-HDACI AND HDAC3/6I SUPPRESSED P. GINGIVALIS-INDUCED EXPRESSION OF IL1B, CCL2, CCL5, CXCL10, MMP1, AND MMP3 IN GFS FROM PATIENTS WITH PERIODONTITIS. OUR RESULTS IDENTIFY HDAC3 AS AN IMPORTANT REGULATOR OF INFLAMMATORY GENE EXPRESSION IN GFS AND SUGGEST THAT THERAPEUTIC TARGETING OF HDAC ACTIVITY, IN PARTICULAR HDAC3, MAY BE CLINICALLY BENEFICIAL IN SUPPRESSING INFLAMMATION IN PERIODONTAL DISEASE. 2020 3 5114 36 PORPHYROMONAS GINGIVALIS LIPOPOLYSACCHARIDE STIMULATION IN HUMAN PERIODONTAL LIGAMENT STEM CELLS: ROLE OF EPIGENETIC MODIFICATIONS TO THE INFLAMMATION. PERIODONTITIS IS A CHRONIC ORAL INFLAMMATORY DISEASE PRODUCED BY BACTERIA. GINGIVAL RETRACTION AND BONE AND CONNECTIVE TISSUES RESORPTION ARE THE HALLMARKS OF THIS DISEASE. CHRONIC PERIODONTITIS MAY CONTRIBUTE TO THE RISK OF ONSET OR PROGRESSION OF NEUROINFLAMMATORY PATHOLOGICAL CONDITIONS, SUCH AS ALZHEIMER'S DISEASE. THE MAIN GOAL OF THE PRESENT STUDY WAS TO INVESTIGATE IF THE ROLE OF EPIGENETIC MODULATIONS IS INVOLVED IN PERIODONTITIS USING HUMAN PERIODONTAL LIGAMENT STEM CELLS (HPDLSCS) AS AN IN VITRO MODEL SYSTEM. HPDLSCS WERE TREATED WITH LIPOPOLYSACCHARIDE OF PORPHYROMONAS GINGIVALIS AND THE EXPRESSION OF PROTEINS ASSOCIATED WITH DNA METHYLATION AND HISTONE ACETYLATION, SUCH AS DNMT1 AND P300, RESPECTIVELY, AND INFLAMMATORY TRANSCRIPTION FACTOR NF-KB, WERE EXAMINED. IMMUNOFLUORESCENCE, WESTERN BLOT AND NEXT GENERATION SEQUENCING RESULTS DEMONSTRATED THAT P. GINGIVALIS LIPOPOLYSACCHARIDE SIGNIFICANTLY REDUCED DNA METHYLASE DNMT1, WHILE IT MARKEDLY UPREGULATED THE LEVEL OF HISTONE ACETYLTRANSFERASE P300 AND NF-KB IN HPDLSCS. OUR RESULTS SHOWED THAT P. GINGIVALIS LIPOPOLYSACCHARIDE MARKEDLY REGULATE THE GENES INVOLVED IN EPIGENETIC MECHANISM, WHICH MAY RESULT IN INFLAMMATION INDUCTION. WE PROPOSE THAT P. GINGIVALIS LIPOPOLYSACCHARIDE-TREATED HPDLSCS COULD BE A POTENTIAL IN VITRO MODEL SYSTEM TO STUDY EPIGENETICS MODULATIONS ASSOCIATED WITH PERIODONTITIS, WHICH MIGHT BE HELPFUL TO IDENTIFY NOVEL BIOMARKERS LINKED TO THIS ORAL INFLAMMATORY DISEASE. 2017 4 592 39 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 5 4582 34 N-TERMINAL BET BROMODOMAIN INHIBITORS DISRUPT A BRD4-P65 INTERACTION AND REDUCE INDUCIBLE NITRIC OXIDE SYNTHASE TRANSCRIPTION IN PANCREATIC BETA-CELLS. CHRONIC INFLAMMATION OF PANCREATIC ISLETS IS A KEY DRIVER OF BETA-CELL DAMAGE THAT CAN LEAD TO AUTOREACTIVITY AND THE EVENTUAL ONSET OF AUTOIMMUNE DIABETES (T1D). IN THE ISLET, ELEVATED LEVELS OF PROINFLAMMATORY CYTOKINES INDUCE THE TRANSCRIPTION OF THE INDUCIBLE NITRIC OXIDE SYNTHASE (INOS) GENE, NOS2, ULTIMATELY RESULTING IN INCREASED NITRIC OXIDE (NO). EXCESSIVE OR PROLONGED EXPOSURE TO NO CAUSES BETA-CELL DYSFUNCTION AND FAILURE ASSOCIATED WITH DEFECTS IN MITOCHONDRIAL RESPIRATION. RECENT STUDIES SHOWED THAT INHIBITION OF THE BROMODOMAIN AND EXTRATERMINAL DOMAIN (BET) FAMILY OF PROTEINS, A DRUGGABLE CLASS OF EPIGENETIC READER PROTEINS, PREVENTS THE ONSET AND PROGRESSION OF T1D IN THE NON-OBESE DIABETIC MOUSE MODEL. WE HYPOTHESIZED THAT BET PROTEINS CO-ACTIVATE TRANSCRIPTION OF CYTOKINE-INDUCED INFLAMMATORY GENE TARGETS IN BETA-CELLS AND THAT SELECTIVE, CHEMOTHERAPEUTIC INHIBITION OF BET BROMODOMAINS COULD REDUCE SUCH TRANSCRIPTION. HERE, WE INVESTIGATED THE ABILITY OF BET BROMODOMAIN SMALL MOLECULE INHIBITORS TO REDUCE THE BETA-CELL RESPONSE TO THE PROINFLAMMATORY CYTOKINE INTERLEUKIN 1 BETA (IL-1BETA). BET BROMODOMAIN INHIBITION ATTENUATED IL-1BETA-INDUCED TRANSCRIPTION OF THE INFLAMMATORY MEDIATOR NOS2 AND CONSEQUENT INOS PROTEIN AND NO PRODUCTION. REDUCED NOS2 TRANSCRIPTION IS CONSISTENT WITH INHIBITION OF NF-KAPPAB FACILITATED BY DISRUPTING THE INTERACTION OF A SINGLE BET FAMILY MEMBER, BRD4, WITH THE NF-KAPPAB SUBUNIT, P65. USING RECENTLY REPORTED SELECTIVE INHIBITORS OF THE FIRST AND SECOND BET BROMODOMAINS, INHIBITION OF ONLY THE FIRST BROMODOMAIN WAS NECESSARY TO REDUCE THE INTERACTION OF BRD4 WITH P65 IN BETA-CELLS. MOREOVER, INHIBITION OF THE FIRST BROMODOMAIN WAS SUFFICIENT TO MITIGATE IL-1BETA-DRIVEN DECREASES IN MITOCHONDRIAL OXYGEN CONSUMPTION RATES AND BETA-CELL VIABILITY. BY IDENTIFYING A ROLE FOR THE INTERACTION BETWEEN BRD4 AND P65 IN CONTROLLING THE RESPONSE OF BETA-CELLS TO PROINFLAMMATORY CYTOKINES, WE PROVIDE MECHANISTIC INFORMATION ON HOW BET BROMODOMAIN INHIBITION CAN DECREASE INFLAMMATION. THESE STUDIES ALSO SUPPORT THE POTENTIAL THERAPEUTIC APPLICATION OF MORE SELECTIVE BET BROMODOMAIN INHIBITORS IN ATTENUATING BETA-CELL INFLAMMATION. 2022 6 194 35 ACETYLSHIKONIN SUPPRESSES INVASION OF PORPHYROMONAS GINGIVALIS?INFECTED YD10B ORAL CANCER CELLS BY MODULATING THE INTERLEUKIN-8/MATRIX METALLOPROTEINASE AXIS. THE DEVELOPMENT OF PHARMACEUTICAL AGENTS POSSESSING ANTI?INVASIVE AND ANTI?METASTATIC ABILITIES, AS WELL AS APOPTOTIC ACTIVITY, IS IMPORTANT IN DECREASING THE INCIDENCE AND RECURRENCE OF ORAL CANCER. CANCER CELLS ARE KNOWN TO ACQUIRE INVASIVENESS NOT ONLY THROUGH EPIGENETIC CHANGES, BUT ALSO FROM INFLAMMATORY STIMULI WITHIN THE TUMOR MICROENVIRONMENT. ACCORDINGLY, THE IDENTIFICATION OF AGENTS THAT CAN SUPPRESS THE INFLAMMATION?PROMOTED INVASIVENESS OF CANCER CELLS MAY BE IMPORTANT IN TREATING CANCER AND IMPROVING THE PROGNOSIS OF PATIENTS WITH CANCER. ACETYLSHIKONIN, A FLAVONOID WITH ANTI?INFLAMMATORY ACTIVITY, INHIBITS PROLIFERATION AND INDUCES APOPTOSIS OF ORAL CANCER CELLS. IN THE PRESENT STUDY, THE ANTI?INVASIVE EFFECT OF ACETYLSHIKONIN ON YD10B ORAL CANCER CELLS INFECTED WITH PORPHYROMONAS GINGIVALIS, A MAJOR PATHOGEN OF CHRONIC PERIODONTITIS, AND THE MECHANISMS INVOLVED WERE INVESTIGATED. FIRSTLY, WE EXAMINED WHETHER P. GINGIVALIS INFECTION INCREASED THE INVASIVENESS OF YD10B CELLS. RESULTS SUGGESTED THAT YD10B ORAL CANCER CELLS BECOME MORE AGGRESSIVE WHEN THEY ARE INFECTED WITH P. GINGIVALIS. SECONDLY, ACETYLSHIKONIN SIGNIFICANTLY INHIBITED THE INVASION OF P. GINGIVALIS?INFECTED YD10B CELLS BY SUPPRESSING IL?8 RELEASE AND IL?8?DEPENDENT MMP RELEASE. THESE DATA SUGGEST THAT ACETYLSHIKONIN MAY BE A USEFUL PREVENTIVE AND THERAPEUTIC CANDIDATE FOR ORAL CANCER THAT IS CHRONICALLY INFECTED WITH PERIODONTAL PATHOGENS. 2018 7 3527 29 IL-6 ENHANCES THE NUCLEAR TRANSLOCATION OF DNA CYTOSINE-5-METHYLTRANSFERASE 1 (DNMT1) VIA PHOSPHORYLATION OF THE NUCLEAR LOCALIZATION SEQUENCE BY THE AKT KINASE. THE EPIGENETIC PROGRAMMING OF GENOMIC DNA IS ACCOMPLISHED, IN PART, BY SEVERAL DNA CYTOSINE-5-METHYLTRANSFERASES THAT ACT BY COVALENTLY MODIFYING CYTOSINES WITH THE ADDITION OF A METHYL GROUP. THIS COVALENT MODIFICATION IS MAINTAINED BY THE DNA CYTOSINE-5-METHYLTRANSFERASE-1 ENZYME (DNMT1), WHICH IS CAPABLE OF ACTING IN CONCERT WITH OTHER SIMILAR ENZYMES TO SILENCE IMPORTANT TUMOR SUPPRESSOR GENES. IL-6 IS A MULTIFUNCTIONAL MEDIATOR OF INFLAMMATION, ACTING THROUGH SEVERAL MAJOR SIGNALING CASCADES, INCLUDING THE PHOSPHATIDYLINOSITOL-3-KINASE PATHWAY (PI-3-K), WHICH ACTIVATES PROTEIN KINASE B (AKT/PKB) DOWNSTREAM. HERE, WE SHOW THAT THE SUBCELLULAR LOCALIZATION OF DNMT1 CAN BE ALTERED BY THE ADDITION OF IL-6, INCREASING THE RATE OF NUCLEAR TRANSLOCATION OF THE ENZYME FROM THE CYTOSOLIC COMPARTMENT. THE MECHANISM OF NUCLEAR TRANSLOCATION OF DNMT1 IS GREATLY ENHANCED BY PHOSPHORYLATION OF THE DNMT1 NUCLEAR LOCALIZATION SIGNAL (NLS) BY PKB/AKT KINASE. MUTAGENIC ALTERATION OF THE TWO AKT TARGET AMINO ACIDS WITHIN THE NLS RESULTS IN A MAJOR LOSS OF DNMT1 NUCLEAR TRANSLOCATION, WHILE THE CREATION OF A "PHOSPHO-MIMIC" AMINO ACID (MUTATION TO ACIDIC RESIDUES) RESTORES THIS COMPARTMENTATION ABILITY. THESE OBSERVATIONS SUGGEST AN INTERESTING HYPOTHESIS REGARDING HOW MEDIATORS OF CHRONIC INFLAMMATION MAY DISTURB THE DELICATE BALANCE OF CELLULAR COMPARTMENTALIZATION OF IMPORTANT PROTEINS, AND REVEALS A POTENTIAL MECHANISM FOR THE INDUCTION OR ENHANCEMENT OF TUMOR GROWTH VIA ALTERATION OF THE COMPONENTS INVOLVED IN THE EPIGENETIC PROGRAMMING OF A CELL. 2007 8 593 45 BET PROTEIN INHIBITION REGULATES CYTOKINE PRODUCTION AND PROMOTES NEUROPROTECTION AFTER SPINAL CORD INJURY. BACKGROUND: SPINAL CORD INJURY (SCI) USUALLY CAUSES A DEVASTATING LIFELONG DISABILITY FOR PATIENTS. AFTER A TRAUMATIC LESION, DISRUPTION OF THE BLOOD-SPINAL CORD BARRIER INDUCES THE INFILTRATION OF MACROPHAGES INTO THE LESION SITE AND THE ACTIVATION OF RESIDENT GLIAL CELLS, WHICH RELEASE CYTOKINES AND CHEMOKINES. THESE EVENTS RESULT IN A PERSISTENT INFLAMMATION, WHICH HAS BOTH DETRIMENTAL AND BENEFICIAL EFFECTS, BUT EVENTUALLY LIMITS FUNCTIONAL RECOVERY AND CONTRIBUTES TO THE APPEARANCE OF NEUROPATHIC PAIN. BROMODOMAIN AND EXTRA-TERMINAL DOMAIN (BET) PROTEINS ARE EPIGENETIC READERS THAT REGULATE THE EXPRESSION OF INFLAMMATORY GENES BY INTERACTING WITH ACETYLATED LYSINE RESIDUES. WHILE BET INHIBITORS ARE A PROMISING THERAPEUTIC STRATEGY FOR CANCER, LITTLE IS KNOWN ABOUT THEIR IMPLICATION AFTER SCI. THUS, THE CURRENT STUDY WAS AIMED TO INVESTIGATE THE ANTI-INFLAMMATORY ROLE OF BET INHIBITORS IN THIS PATHOLOGIC CONDITION. METHODS: WE EVALUATED THE EFFECTIVENESS OF THE BET INHIBITOR JQ1 TO MODIFY MACROPHAGE REACTIVITY IN VITRO AND TO MODULATE INFLAMMATION IN A SCI MICE MODEL. WE ANALYZED THE EFFECTS OF BET INHIBITION IN PRO-INFLAMMATORY AND ANTI-INFLAMMATORY CYTOKINE PRODUCTION IN VITRO AND IN VIVO. WE DETERMINED THE EFFECTIVENESS OF BET INHIBITION IN TISSUE SPARING, INFLAMMATION, NEURONAL PROTECTION, AND BEHAVIORAL OUTCOME AFTER SCI. RESULTS: WE HAVE FOUND THAT THE BET INHIBITOR JQ1 REDUCED THE LEVELS OF PRO-INFLAMMATORY MEDIATORS AND INCREASED THE EXPRESSION OF ANTI-INFLAMMATORY CYTOKINES. A PROLONGED TREATMENT WITH JQ1 ALSO DECREASED REACTIVITY OF MICROGLIA/MACROPHAGES, ENHANCED NEUROPROTECTION AND FUNCTIONAL RECOVERY, AND ACUTELY REDUCED NEUROPATHIC PAIN AFTER SCI. CONCLUSIONS: BET PROTEIN INHIBITION IS AN EFFECTIVE TREATMENT TO REGULATE CYTOKINE PRODUCTION AND PROMOTE NEUROPROTECTION AFTER SCI. THESE NOVEL RESULTS DEMONSTRATE FOR THE FIRST TIME THAT TARGETING BET PROTEINS IS AN ENCOURAGING APPROACH FOR SCI REPAIR AND A POTENTIAL STRATEGY TO TREAT OTHER INFLAMMATORY PATHOLOGIES. 2019 9 5088 39 PIPERLONGUMINE REGULATES EPIGENETIC MODULATION AND ALLEVIATES PSORIASIS-LIKE SKIN INFLAMMATION VIA INHIBITION OF HYPERPROLIFERATION AND INFLAMMATION. PSORIASIS IS AN AUTOIMMUNE SKIN DISEASE, WHERE CHRONIC IMMUNE RESPONSES DUE TO EXAGGERATED CYTOKINE SIGNALING, ABNORMAL DIFFERENTIATION, AND EVASION OF KERATINOCYTES APOPTOSIS PLAYS A CRUCIAL ROLE IN MEDIATING ABNORMAL KERATINOCYTES HYPERPROLIFERATION. FROM THE THERAPEUTIC PERSPECTIVE, THE MOLECULES WITH STRONG ANTI-PROLIFERATIVE AND ANTI-INFLAMMATORY PROPERTIES COULD HAVE TREMENDOUS RELEVANCE. IN THIS STUDY, WE DEMONSTRATED THAT PIPERLONGUMINE (PPL) TREATMENT EFFECTIVELY ABROGATED THE HYPERPROLIFERATION AND DIFFERENTIATION OF KERATINOCYTES BY INDUCING ROS-MEDIATED LATE APOPTOSIS WITH LOSS OF MITOCHONDRIAL MEMBRANE POTENTIAL. BESIDES, THE ARREST OF CELL CYCLE WAS FOUND AT SUB-G1 PHASE AS A RESULT OF DNA FRAGMENTATION. MOLECULARLY, INHIBITION OF STAT3 AND AKT SIGNALING WAS OBSERVED WITH A DECREASE IN PROLIFERATIVE MARKERS SUCH AS PCNA, KI67, AND CYCLIN D1 ALONG WITH ANTI-APOPTOTIC BCL-2 PROTEIN EXPRESSION. KERATIN 17 IS A CRITICAL REGULATOR OF KERATINOCYTE DIFFERENTIATION, AND IT WAS FOUND TO BE DOWNREGULATED WITH PPL SIGNIFICANTLY. FURTHERMORE, PROMINENT ANTI-INFLAMMATORY EFFECTS WERE OBSERVED BY INHIBITION OF LIPOPOLYSACCHARIDE (LPS)/IMIQUIMOD (IMQ)-INDUCED P65 NF-KAPPAB SIGNALING CASCADE AND STRONGLY INHIBITED THE PRODUCTION OF CYTOKINE STORM INVOLVED IN PSORIASIS-LIKE SKIN INFLAMMATION, THUS LED TO THE RESTORATION OF NORMAL EPIDERMAL ARCHITECTURE WITH REDUCTION OF EPIDERMAL HYPERPLASIA AND SPLENOMEGALY. IN ADDITION, PPL EPIGENETICALLY INHIBITED HISTONE-MODIFYING ENZYMES, WHICH INCLUDE HISTONE DEACETYLASES (HDACS) OF CLASS I (HDAC1-4) AND CLASS II (HDAC6) EVALUATED BY IMMUNOBLOTTING AND HDAC ENZYME ASSAY KIT. IN ADDITION, OUR RESULTS SHOW THAT PPL EFFECTIVELY INHIBITS THE NUCLEAR TRANSLOCATION OF P65 AND A HISTONE MODULATOR HDAC3, THUS SEQUESTERED IN THE CYTOPLASM OF MACROPHAGES. FURTHERMORE, PPL EFFECTIVELY ENHANCED THE PROTEIN-PROTEIN INTERACTIONS OF HDAC3 AND P65 WITH IKAPPABALPHA, WHICH WAS DISRUPTED BY LPS STIMULATION AND WERE EVALUATED BY CO-IP AND MOLECULAR MODELING. COLLECTIVELY, OUR FINDINGS INDICATE THAT PIPERLONGUMINE MAY SERVE AS AN ANTI-PROLIFERATIVE AND ANTI-INFLAMMATORY AGENT AND COULD SERVE AS A POTENTIAL THERAPEUTIC OPTION IN TREATING PSORIASIS. 2020 10 594 25 BET PROTEIN INHIBITOR JQ1 MODULATES MITOCHONDRIAL DYSFUNCTION AND OXIDATIVE STRESS INDUCED BY CHRONIC KIDNEY DISEASE. AMONG THE MECHANISMS INVOLVED IN THE PROGRESSION OF KIDNEY DISEASE, MITOCHONDRIAL DYSFUNCTION HAS SPECIAL RELEVANCE. EPIGENETIC DRUGS SUCH AS INHIBITORS OF EXTRA-TERMINAL DOMAIN PROTEINS (IBET) HAVE SHOWN BENEFICIAL EFFECTS IN EXPERIMENTAL KIDNEY DISEASE, MAINLY BY INHIBITING PROLIFERATIVE AND INFLAMMATORY RESPONSES. THE IMPACT OF IBET ON MITOCHONDRIAL DAMAGE WAS EXPLORED IN IN VITRO STUDIES IN RENAL CELLS STIMULATED WITH TGF-BETA1 AND IN VIVO IN MURINE UNILATERAL URETERAL OBSTRUCTION (UUO) MODEL OF PROGRESSIVE KIDNEY DAMAGE. IN VITRO, JQ1 PRETREATMENT PREVENTED THE TGF-BETA1-INDUCED DOWNREGULATION OF COMPONENTS OF THE OXIDATIVE PHOSPHORYLATION CHAIN (OXPHOS), SUCH AS CYTOCHROME C AND CV-ATP5A IN HUMAN PROXIMAL TUBULAR CELLS. IN ADDITION, JQ1 ALSO PREVENTED THE ALTERED MITOCHONDRIAL DYNAMICS BY AVOIDING THE INCREASE IN THE DRP-1 FISSION FACTOR. IN UUO MODEL, RENAL GENE EXPRESSION LEVELS OF CYTOCHROME C AND CV-ATP5A AS WELL AS PROTEIN LEVELS OF CYTOCHROME C WERE REDUCED THESE CHANGES WERE PREVENTED BY JQ1 ADMINISTRATION. IN ADDITION, JQ1 DECREASED PROTEIN LEVELS OF THE DRP1 FISSION PROTEIN AND INCREASED THE OPA-1 FUSION PROTEIN, RESTORING MITOCHONDRIAL DYNAMICS. MITOCHONDRIA ALSO PARTICIPATE IN THE MAINTENANCE OF REDOX BALANCE. JQ1 RESTORED THE GENE EXPRESSION OF ANTIOXIDANT PROTEINS, SUCH AS CATALASE AND HEME OXYGENASE 1 IN TGF-BETA1-STIMULATED HUMAN PROXIMAL TUBULAR CELLS AND IN MURINE OBSTRUCTED KIDNEYS. INDEED, IN TUBULAR CELLS, JQ1 DECREASED ROS PRODUCTION INDUCED BY STIMULATION WITH TGF-BETA1, AS EVALUATED BY MITOSOXTM. IBETS, SUCH AS JQ1, IMPROVE MITOCHONDRIAL DYNAMICS, FUNCTIONALITY, AND OXIDATIVE STRESS IN KIDNEY DISEASE. 2023 11 2228 32 EPIGENETIC MODIFICATIONS OF HISTONES IN PERIODONTAL DISEASE. PERIODONTITIS IS A CHRONIC INFECTIOUS DISEASE DRIVEN BY DYSBIOSIS, AN IMBALANCE BETWEEN COMMENSAL BACTERIA AND THE HOST ORGANISM. PERIODONTITIS IS A LEADING CAUSE OF TOOTH LOSS IN ADULTS AND OCCURS IN ABOUT 50% OF THE US POPULATION. IN ADDITION TO THE CLINICAL CHALLENGES ASSOCIATED WITH TREATING PERIODONTITIS, THE PROGRESSION AND CHRONIC NATURE OF THIS DISEASE SERIOUSLY AFFECT HUMAN HEALTH. EMERGING EVIDENCE SUGGESTS THAT PERIODONTITIS IS ASSOCIATED WITH MECHANISMS BEYOND BACTERIA-INDUCED PROTEIN AND TISSUE DEGRADATION. HERE, WE HYPOTHESIZE THAT BACTERIA ARE ABLE TO INDUCE EPIGENETIC MODIFICATIONS IN ORAL EPITHELIAL CELLS MEDIATED BY HISTONE MODIFICATIONS. IN THIS STUDY, WE FOUND THAT DYSBIOSIS IN VIVO LED TO EPIGENETIC MODIFICATIONS, INCLUDING ACETYLATION OF HISTONES AND DOWNREGULATION OF DNA METHYLTRANSFERASE 1. IN ADDITION, IN VITRO EXPOSURE OF ORAL EPITHELIAL CELLS TO LIPOPOLYSACCHARIDES RESULTED IN HISTONE MODIFICATIONS, ACTIVATION OF TRANSCRIPTIONAL COACTIVATORS, SUCH AS P300/CBP, AND ACCUMULATION OF NUCLEAR FACTOR-KAPPAB (NF-KAPPAB). GIVEN THAT ORAL EPITHELIAL CELLS ARE THE FIRST LINE OF DEFENSE FOR THE PERIODONTIUM AGAINST BACTERIA, WE ALSO EVALUATED WHETHER ACTIVATION OF PATHOGEN RECOGNITION RECEPTORS INDUCED HISTONE MODIFICATIONS. WE FOUND THAT ACTIVATION OF THE TOLL-LIKE RECEPTORS 1, 2, AND 4 AND THE NUCLEOTIDE-BINDING OLIGOMERIZATION DOMAIN PROTEIN 1 INDUCED HISTONE ACETYLATION IN ORAL EPITHELIAL CELLS. OUR FINDINGS CORROBORATE THE EMERGING CONCEPT THAT EPIGENETIC MODIFICATIONS PLAY A ROLE IN THE DEVELOPMENT OF PERIODONTITIS. 2016 12 698 37 BROMODOMAIN CONTAINING PROTEIN 4 (BRD4) REGULATES EXPRESSION OF ITS INTERACTING COACTIVATORS IN THE INNATE RESPONSE TO RESPIRATORY SYNCYTIAL VIRUS. BROMODOMAIN-CONTAINING PROTEIN 4 PLAYS A CENTRAL ROLE IN COORDINATING THE COMPLEX EPIGENETIC COMPONENT OF THE INNATE IMMUNE RESPONSE. PREVIOUS STUDIES IMPLICATED BRD4 AS A COMPONENT OF A CHROMATIN-MODIFYING COMPLEX THAT IS DYNAMICALLY RECRUITED TO A NETWORK OF PROTECTIVE CYTOKINES BY BINDING ACTIVATED TRANSCRIPTION FACTORS, POLYMERASES, AND HISTONES TO TRIGGER THEIR RAPID EXPRESSION VIA TRANSCRIPTIONAL ELONGATION. OUR PREVIOUS STUDY EXTENDED OUR UNDERSTANDING OF THE AIRWAY EPITHELIAL BRD4 INTERACTOME BY IDENTIFYING OVER 100 FUNCTIONALLY IMPORTANT COACTIVATORS AND TRANSCRIPTION FACTORS, WHOSE ASSOCIATION IS INDUCED BY RESPIRATORY SYNCYTIAL VIRUS (RSV) INFECTION. RSV IS AN ETIOLOGICAL AGENT OF RECURRENT RESPIRATORY TRACT INFECTIONS ASSOCIATED WITH EXACERBATIONS OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE. USING A HIGHLY SELECTIVE SMALL-MOLECULE BRD4 INHIBITOR (ZL0454) DEVELOPED BY US, WE EXTEND THESE FINDINGS TO IDENTIFY THE GENE REGULATORY NETWORK DEPENDENT ON BRD4 BROMODOMAIN (BD) INTERACTIONS. HUMAN SMALL AIRWAY EPITHELIAL CELLS WERE INFECTED IN THE ABSENCE OR PRESENCE OF ZL0454, AND GENE EXPRESSION PROFILING WAS PERFORMED. A HIGHLY REPRODUCIBLE DATASET WAS OBTAINED WHICH INDICATED THAT BRD4 MEDIATES BOTH ACTIVATION AND REPRESSION OF RSV-INDUCIBLE GENE REGULATORY NETWORKS CONTROLLING CYTOKINE EXPRESSION, INTERFERON (IFN) PRODUCTION, AND EXTRACELLULAR MATRIX REMODELING. INDEX GENES OF FUNCTIONALLY SIGNIFICANT CLUSTERS WERE VALIDATED INDEPENDENTLY. WE DISCOVER THAT BRD4 REGULATES THE EXPRESSION OF ITS OWN GENE DURING THE INNATE IMMUNE RESPONSE. INTERESTINGLY, BRD4 ACTIVATES THE EXPRESSION OF NFKAPPAB/RELA, A COACTIVATOR THAT BINDS TO BRD4 IN A BD-DEPENDENT MANNER. WE EXTEND THIS FINDING TO SHOW THAT BRD4 ALSO REGULATES OTHER COMPONENTS OF ITS FUNCTIONAL INTERACTOME, INCLUDING THE MEDIATOR (MED) COACTIVATOR COMPLEX AND THE SWI/SNF-RELATED, MATRIX-ASSOCIATED, ACTIN-DEPENDENT REGULATOR OF CHROMATIN (SMARC) SUBUNITS. TO PROVIDE FURTHER INSIGHT INTO MECHANISMS FOR BRD4 IN RSV EXPRESSION, WE MAPPED 7,845 RSV-INDUCIBLE TN5 TRANSPOSASE PEAKS ONTO THE BRD4-DEPENDENT GENE BODIES. THESE WERE LOCATED IN PROMOTERS AND INTRONS OF CYTOSTRUCTURAL AND EXTRACELLULAR MATRIX (ECM) FORMATION GENES. THESE DATA INDICATE THAT BRD4 MEDIATES THE DYNAMIC RESPONSE OF AIRWAY EPITHELIAL CELLS TO RNA INFECTION BY MODULATING THE EXPRESSION OF ITS COACTIVATORS, CONTROLLING THE EXPRESSION OF HOST DEFENSE MECHANISMS AND REMODELING GENES THROUGH CHANGES IN PROMOTER ACCESSIBILITY. 2021 13 689 36 BRD4 AS A THERAPEUTIC TARGET IN PULMONARY DISEASES. BROMODOMAIN AND EXTRA-TERMINAL DOMAIN (BET) PROTEINS ARE EPIGENETIC MODULATORS THAT REGULATE GENE TRANSCRIPTION THROUGH INTERACTING WITH ACETYLATED LYSINE RESIDUES OF HISTONE PROTEINS. BET PROTEINS HAVE MULTIPLE ROLES IN REGULATING KEY CELLULAR FUNCTIONS SUCH AS CELL PROLIFERATION, DIFFERENTIATION, INFLAMMATION, OXIDATIVE AND REDOX BALANCE, AND IMMUNE RESPONSES. AS A RESULT, BET PROTEINS HAVE BEEN FOUND TO BE ACTIVELY INVOLVED IN A BROAD RANGE OF HUMAN LUNG DISEASES INCLUDING ACUTE LUNG INFLAMMATION, ASTHMA, PULMONARY ARTERIAL HYPERTENSION, PULMONARY FIBROSIS, AND CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD). DUE TO THE IDENTIFICATION OF SPECIFIC SMALL MOLECULAR INHIBITORS OF BET PROTEINS, TARGETING BET IN THESE LUNG DISEASES HAS BECOME AN AREA OF INCREASING INTEREST. EMERGING EVIDENCE HAS DEMONSTRATED THE BENEFICIAL EFFECTS OF BET INHIBITORS IN PRECLINICAL MODELS OF VARIOUS HUMAN LUNG DISEASES. THIS IS, IN GENERAL, LARGELY RELATED TO THE ABILITY OF BET PROTEINS TO BIND TO PROMOTERS OF GENES THAT ARE CRITICAL FOR INFLAMMATION, DIFFERENTIATION, AND BEYOND. BY MODULATING THESE CRITICAL GENES, BET PROTEINS ARE INTEGRATED INTO THE PATHOGENESIS OF DISEASE PROGRESSION. THE INTRINSIC HISTONE ACETYLTRANSFERASE ACTIVITY OF BROMODOMAIN-CONTAINING PROTEIN 4 (BRD4) IS OF PARTICULAR INTEREST, SEEMS TO ACT INDEPENDENTLY OF ITS BROMODOMAIN BINDING ACTIVITY, AND HAS IMPLICATION IN SOME CONTEXTS. IN THIS REVIEW, WE PROVIDE A BRIEF OVERVIEW OF THE RESEARCH ON BET PROTEINS WITH A FOCUS ON BRD4 IN SEVERAL MAJOR HUMAN LUNG DISEASES, THE UNDERLYING MOLECULAR MECHANISMS, AS WELL AS FINDINGS OF TARGETING BET PROTEINS USING PHARMACEUTICAL INHIBITORS IN DIFFERENT LUNG DISEASES PRECLINICALLY. 2023 14 5995 33 TGFBETA-INDUCED FIBROBLAST ACTIVATION REQUIRES PERSISTENT AND TARGETED HDAC-MEDIATED GENE REPRESSION. TISSUE FIBROSIS IS A CHRONIC DISEASE DRIVEN BY PERSISTENT FIBROBLAST ACTIVATION THAT HAS RECENTLY BEEN LINKED TO EPIGENETIC MODIFICATIONS. HERE, WE SCREENED A SMALL LIBRARY OF EPIGENETIC SMALL-MOLECULE MODULATORS TO IDENTIFY COMPOUNDS CAPABLE OF INHIBITING OR REVERSING TGFBETA-MEDIATED FIBROBLAST ACTIVATION. WE IDENTIFIED PRACINOSTAT, AN HDAC INHIBITOR, AS A POTENT ATTENUATOR OF LUNG FIBROBLAST ACTIVATION AND CONFIRMED ITS EFFICACY IN PATIENT-DERIVED FIBROBLASTS ISOLATED FROM FIBROTIC LUNG TISSUE. MECHANISTICALLY, WE FOUND THAT HDAC-DEPENDENT TRANSCRIPTIONAL REPRESSION WAS AN EARLY AND ESSENTIAL EVENT IN TGFBETA-MEDIATED FIBROBLAST ACTIVATION. TREATMENT OF LUNG FIBROBLASTS WITH PRACINOSTAT BROADLY ATTENUATED TGFBETA-MEDIATED EPIGENETIC REPRESSION AND PROMOTED FIBROBLAST QUIESCENCE. WE CONFIRMED A SPECIFIC ROLE FOR HDAC-DEPENDENT HISTONE DEACETYLATION IN THE PROMOTER REGION OF THE ANTI-FIBROTIC GENE PPARGC1A (PGC1ALPHA) IN RESPONSE TO TGFBETA STIMULATION. FINALLY, WE IDENTIFIED HDAC7 AS A KEY FACTOR WHOSE SIRNA-MEDIATED KNOCKDOWN ATTENUATES FIBROBLAST ACTIVATION WITHOUT ALTERING GLOBAL HISTONE ACETYLATION. TOGETHER, THESE RESULTS PROVIDE NOVEL MECHANISTIC INSIGHT INTO THE ESSENTIAL ROLE HDACS PLAY IN TGFBETA-MEDIATED FIBROBLAST ACTIVATION VIA TARGETED GENE REPRESSION. 2019 15 4302 41 MICRORNA-223 CONTROLS THE EXPRESSION OF HISTONE DEACETYLASE 2: A NOVEL AXIS IN COPD. REDUCED ACTIVITY OF HISTONE DEACETYLASE 2 (HDAC2) HAS BEEN DESCRIBED IN PATIENTS WITH CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD), BUT THE MECHANISMS RESULTING IN DECREASED EXPRESSION OF THIS IMPORTANT EPIGENETIC MODIFIER REMAIN UNKNOWN. HERE, WE EMPLOYED SEVERAL IN VITRO EXPERIMENTS TO ADDRESS THE ROLE OF MICRORNAS (MIRNAS) ON THE REGULATION OF HDAC2 IN ENDOTHELIAL CELLS. MANIPULATION OF MIRNA LEVELS IN HUMAN PULMONARY ARTERY ENDOTHELIAL CELLS (HPAEC) WAS ACHIEVED BY USING ELECTROPORATION WITH ANTI-MIRNAS AND MIRNA MIMICS. TARGET PREDICTION SOFTWARE IDENTIFIED MIR-223 AS A POTENTIAL REPRESSOR OF HDAC2. IN SUBSEQUENT STIMULATION EXPERIMENTS USING INFLAMMATORY CYTOKINES KNOWN TO BE INCREASED IN PATIENTS WITH COPD, MIR-223 WAS FOUND TO BE SIGNIFICANTLY INDUCED. FUNCTIONAL ANALYSIS DEMONSTRATED THAT OVEREXPRESSION OF MIR-223 DECREASED HDAC2 EXPRESSION AND ACTIVITY IN HPAEC. CONVERSELY, HDAC2 EXPRESSION AND ACTIVITY WAS PRESERVED IN ANTI-MIR-223-TREATED CELLS. DIRECT MIRNA-TARGET INTERACTION WAS CONFIRMED BY REPORTER GENE ASSAY. IN A NEXT STEP, REDUCED EXPRESSION OF HDAC2 WAS FOUND TO INCREASE THE LEVELS OF THE CHEMOKINE FRACTALKINE (CX3CL1). IN VIVO STUDIES CONFIRMED ELEVATED EXPRESSION LEVELS OF MIR-223 IN MICE EXPOSED TO CIGARETTE SMOKE AND IN EMPHYSEMATOUS LUNG TISSUE FROM LPS-TREATED MICE. MOREOVER, A SIGNIFICANT INVERSE CORRELATION OF MIR-223 AND HDAC2 EXPRESSION WAS FOUND IN TWO INDEPENDENT COHORTS OF COPD PATIENTS. THESE DATA EMPHASIZE THAT MIR-223, THE MOST PREVALENT MIRNA IN COPD, CONTROLS EXPRESSION AND ACTIVITY OF HDAC2 IN PULMONARY CELLS, WHICH, IN TURN, MIGHT ALTER THE EXPRESSION PROFILE OF CHEMOKINES. THIS PATHWAY PROVIDES A NOVEL PATHOGENIC LINK BETWEEN DYSREGULATED MIRNA EXPRESSION AND EPIGENETIC ACTIVITY IN COPD. KEY MESSAGES: HISTONE DEACETYLASE 2 IS DIRECTLY TARGETED BY MIR-223. LEVELS OF MIR-223 ARE INDUCED BY INTERLEUKIN-1BETA AND TUMOR NECROSIS FACTOR-ALPHA. MIR-223 CONTROLS THE EXPRESSION OF FRACTALKINE BY TARGETING HISTONE DEACETYLASE 2. MIR-223 LEVELS ARE INCREASED IN COPD MOUSE MODELS. MIR-223 LEVELS INVERSELY CORRELATE WITH HDAC2 EXPRESSION IN COPD PATIENTS. 2016 16 172 30 ABSENCE OF HDAC3 BY MATRIX STIFFNESS PROMOTES CHROMATIN REMODELING AND FIBROBLAST ACTIVATION IN IDIOPATHIC PULMONARY FIBROSIS. IDIOPATHIC PULMONARY FIBROSIS (IPF) IS A CHRONIC AND FATAL DISEASE CHARACTERIZED BY PROGRESSIVE AND IRREVERSIBLE LUNG SCARRING ASSOCIATED WITH PERSISTENT ACTIVATION OF FIBROBLASTS. EPIGENETICS COULD INTEGRATE DIVERSE MICROENVIRONMENTAL SIGNALS, SUCH AS STIFFNESS, TO DIRECT PERSISTENT FIBROBLAST ACTIVATION. HISTONE MODIFICATIONS BY DEACETYLASES (HDAC) MAY PLAY AN ESSENTIAL ROLE IN THE GENE EXPRESSION CHANGES INVOLVED IN THE PATHOLOGICAL REMODELING OF THE LUNG. PARTICULARLY, HDAC3 IS CRUCIAL FOR MAINTAINING CHROMATIN AND REGULATING GENE EXPRESSION, BUT LITTLE IS KNOWN ABOUT ITS ROLE IN IPF. IN THE STUDY, CONTROL AND IPF-DERIVED FIBROBLASTS WERE USED TO DETERMINE THE INFLUENCE OF HDAC3 ON CHROMATIN REMODELING AND GENE EXPRESSION ASSOCIATED WITH IPF SIGNATURE. ADDITIONALLY, THE CELLS WERE GROWN ON HYDROGELS TO MIMIC THE STIFFNESS OF A FIBROTIC LUNG. OUR RESULTS SHOWED A DECREASED HDAC3 IN THE NUCLEUS OF IPF FIBROBLASTS, WHICH CORRELATES WITH CHANGES IN NUCLEUS SIZE AND HETEROCHROMATIN LOSS. THE INHIBITION OF HDAC3 WITH A PHARMACOLOGICAL INHIBITOR CAUSES HYPERACETYLATION OF H3K9 AND PROVOKES AN INCREASED EXPRESSION OF COL1A1, ACTA2, AND P21. COMPARABLE RESULTS WERE FOUND IN HYDROGELS, WHERE MATRIX STIFFNESS PROMOTES THE LOSS OF NUCLEAR HDAC3 AND INCREASES THE PROFIBROTIC SIGNATURE. FINALLY, LATRUNCULIN B WAS USED TO CONFIRM THAT CHANGES BY STIFFNESS DEPEND ON THE MECHANOTRANSDUCTION SIGNALS. TOGETHER, THESE RESULTS SUGGEST THAT HDAC3 COULD BE A LINK BETWEEN EPIGENETIC MECHANISMS AND THE FIBROTIC MICROENVIRONMENT. 2023 17 2247 26 EPIGENETIC MODULATION OF MACROPHAGE POLARIZATION PREVENTS LUMBAR DISC DEGENERATION. INFLAMMATION PLAYS AN ESSENTIAL ROLE IN THE DEVELOPMENT OF LUMBAR DISC DEGENERATION (LDD), ALTHOUGH THE EXACT EFFECTS OF MACROPHAGE SUBTYPES ON LDD REMAIN UNCLEAR. BASED ON PREVIOUS STUDIES, WE HYPOTHESIZED THAT M2-POLARIZATION OF LOCAL MACROPHAGES AND SIMULTANEOUS SUPPRESSION OF THEIR PRODUCTION OF FIBROTIC TRANSFORMING GROWTH FACTOR BETA 1 (TGFBETA1) COULD INHIBIT PROGRESSION OF LDD. THUS, WE APPLIED AN ORTHOTOPIC INJECTION OF ADENO-ASSOCIATED VIRUS (AAV) CARRYING SHRNA FOR DNA METHYLTRANSFERASE 1 (DNMT1) AND/OR SHRNA FOR TGFBETA1 UNDER A MACROPHAGE-SPECIFIC CD68 PROMOTER TO SPECIFICALLY TARGET LOCAL MACROPHAGES IN A MOUSE MODEL FOR LDD. WE FOUND THAT SHDNMT1 SIGNIFICANTLY REDUCED LEVELS OF THE PRO-INFLAMMATORY CYTOKINES TNFALPHA, IL-1BETA AND IL-6, SIGNIFICANTLY INCREASED LEVELS OF THE ANTI-INFLAMMATORY CYTOKINES IL-4 AND IL-10, SIGNIFICANTLY INCREASED M2 MACROPHAGE POLARIZATION, SIGNIFICANTLY REDUCED CELL APOPTOSIS IN THE DISC DEGENERATION ZONE AND SIGNIFICANTLY REDUCED LDD-ASSOCIATED PAIN. THE ANTI-APOPTOTIC AND ANTI-PAIN EFFECTS WERE FURTHER STRENGTHENED BY CO-APPLICATION OF SHTGFBETA1. TOGETHER, THESE DATA SUGGEST THAT M2 POLARIZATION OF MACROPHAGES INDUCED BY BOTH EPIGENETIC MODULATION AND SUPPRESSED PRODUCTION AND RELEASE OF TGFBETA1 FROM POLARIZED M2 MACROPHAGES, MAY HAVE A DEMONSTRABLE THERAPEUTIC EFFECT ON LDD. 2020 18 446 36 APABETALONE DOWNREGULATES FIBROTIC, INFLAMMATORY AND CALCIFIC PROCESSES IN RENAL MESANGIAL CELLS AND PATIENTS WITH RENAL IMPAIRMENT. EPIGENETIC MECHANISMS ARE IMPLICATED IN TRANSCRIPTIONAL PROGRAMS DRIVING CHRONIC KIDNEY DISEASE (CKD). APABETALONE IS AN ORALLY AVAILABLE INHIBITOR OF BROMODOMAIN AND EXTRATERMINAL (BET) PROTEINS, WHICH ARE EPIGENETIC READERS THAT MODULATE GENE EXPRESSION. IN THE PHASE 3 BETONMACE TRIAL, APABETALONE REDUCED RISK OF MAJOR ADVERSE CARDIAC EVENTS (MACE) BY 50% IN THE CKD SUBPOPULATION, INDICATING FAVORABLE EFFECTS ALONG THE KIDNEY-HEART AXIS. ACTIVATION OF HUMAN RENAL MESANGIAL CELLS (HRMCS) TO A CONTRACTILE PHENOTYPE THAT OVERPRODUCES EXTRACELLULAR MATRIX (ECM) AND INFLAMMATORY CYTOKINES, AND PROMOTES CALCIFICATION, FREQUENTLY ACCOMPANIES CKD TO DRIVE PATHOLOGY. HERE, WE SHOW APABETALONE DOWNREGULATED HRMC ACTIVATION WITH TGF-BETA1 STIMULATION BY SUPPRESSING TGF-BETA1-INDUCED ALPHA-SMOOTH MUSCLE ACTIN (ALPHA-SMA) EXPRESSION, ALPHA-SMA ASSEMBLY INTO STRESS FIBERS, ENHANCED CONTRACTION, COLLAGEN OVERPRODUCTION, AND EXPRESSION OF KEY DRIVERS OF FIBROSIS, INFLAMMATION, OR CALCIFICATION INCLUDING THROMBOSPONDIN, FIBRONECTIN, PERIOSTIN, SPARC, INTERLEUKIN 6, AND ALKALINE PHOSPHATASE. LIPOPOLYSACCHARIDE-STIMULATED EXPRESSION OF INFLAMMATORY GENES IL6, IL1B, AND PTGS2 WAS ALSO SUPPRESSED. TRANSCRIPTOMICS CONFIRMED APABETALONE AFFECTED GENE SETS OF ECM REMODELING AND INTEGRINS. CLINICAL TRANSLATION OF IN VITRO RESULTS WAS INDICATED IN CKD PATIENTS WHERE A SINGLE DOSE OF APABETALONE REDUCED PLASMA LEVELS OF KEY PRO-FIBROTIC AND INFLAMMATORY MARKERS, AND INDICATED INHIBITION OF TGF-BETA1 SIGNALING. WHILE PLASMA PROTEINS CANNOT BE TRACED TO THE KIDNEY ALONE, ANTI-FIBROTIC AND ANTI-INFLAMMATORY EFFECTS OF APABETALONE IDENTIFIED IN THIS STUDY ARE CONSISTENT WITH THE OBSERVED DECREASE IN CARDIOVASCULAR RISK IN CKD PATIENTS. 2023 19 3193 39 HDAC INHIBITION REGULATES OXIDATIVE STRESS IN CD4(+)THELPER CELLS OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE AND NON-SMALL CELL LUNG CANCER PATIENTS VIA MITOCHONDRIAL TRANSCRIPTION FACTOR A (MTTFA) MODULATING NF-KAPPAB/HIF1ALPHA AXIS. HISTONE DEACETYLASES (HDACS) PLAY A CRUCIAL ROLE IN THE EPIGENETIC REGULATION OF GENE EXPRESSION BY REMODELLING CHROMATIN. ISOENZYMES OF THE HDAC FAMILY EXHIBIT ABERRANT REGULATION IN A WIDE VARIETY OF CANCERS AS WELL AS SEVERAL INFLAMMATORY LUNG DISORDERS LIKE CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD). INHIBITION OF HDACS IS A POTENTIAL THERAPEUTIC STRATEGY THAT COULD BE USED TO REVERSE EPIGENETIC MODIFICATION. TRICHOSTATIN A (TSA), A POWERFUL HISTONE DEACETYLASE (HDAC) INHIBITOR, HAS ANTI-CANCER EFFECTS IN NUMEROUS CANCER TYPES. HOWEVER, IT IS NOT YET APPARENT HOW HDAC INHIBITORS AFFECT HUMAN NON-SMALL CELL LUNG CANCER CELLS (NSCLC) AND COPD. THIS STUDY AIMS TO INVESTIGATE TSA'S ROLE IN RESTORING MITOCHONDRIAL DYSFUNCTION AND ITS EFFECT ON HYPOXIA AND INFLAMMATION IN CD4(+)T CELLS OBTAINED FROM PATIENTS WITH COPD AND LUNG CANCER. AS A RESULT OF TREATMENT WITH TSA, THERE IS A REDUCTION IN THE EXPRESSION OF INFLAMMATORY CYTOKINES AND A DECREASED ENRICHMENT OF TRANSCRIPTIONAL FACTORS ASSOCIATED WITH INFLAMMATION AT VEGFA GENE LOCI. WE HAVE SEEN A SUBSTANTIAL DECREASE IN THE EXPRESSION OF NF-KAPPAB AND HIF1ALPHA, WHICH ARE THE CRITICAL MEDIATORS OF INFLAMMATION AND HYPOXIA, RESPECTIVELY. FOLLOWING TSA TREATMENT, MTTFA EXPRESSION WAS INCREASED, FACILITATING PATIENTS WITH COPD AND NSCLC IN THE RECOVERY OF THEIR DYSFUNCTIONAL MITOCHONDRIA. FURTHERMORE, WE HAVE DISCOVERED THAT TSA TREATMENT IN PATIENTS WITH COPD AND NSCLC MAY LEAD TO IMMUNOPROTECTIVE NESS BY INDUCING TH1NESS. OUR FINDING GIVES A NEW INSIGHT INTO THE EXISTING BODY OF KNOWLEDGE REGARDING TSA-BASED THERAPEUTIC METHODS AND HIGHLIGHTS THE NECESSITY OF EPIGENETIC THERAPY FOR THESE DEVASTATING LUNG DISORDERS. 2023 20 3720 32 INHIBITION OF CLASS I HISTONE DEACETYLASES ABROGATES TUMOR GROWTH FACTOR BETA EXPRESSION AND DEVELOPMENT OF FIBROSIS DURING CHRONIC PANCREATITIS. PANCREATIC FIBROSIS IS THE HALLMARK OF CHRONIC PANCREATITIS, A HIGHLY DEBILITATING DISEASE FOR WHICH THERE IS CURRENTLY NO CURE. THE KEY EVENT AT THE BASIS OF PANCREATIC FIBROSIS IS THE DEPOSITION OF EXTRACELLULAR MATRIX PROTEINS BY ACTIVATED PANCREATIC STELLATE CELLS (PSCS). TRANSFORMING GROWTH FACTOR BETA (TGFBETA) IS A POTENT PROFIBROTIC FACTOR IN THE PANCREAS AS IT PROMOTES THE ACTIVATION OF PSC; THUS, PHARMACOLOGIC INTERVENTIONS THAT EFFECTIVELY REDUCE TGFBETA EXPRESSION HARBOR CONSIDERABLE THERAPEUTIC POTENTIAL IN THE TREATMENT OF CHRONIC PANCREATITIS. IN THIS STUDY, WE INVESTIGATED WHETHER TGFBETA EXPRESSION IS REDUCED BY PHARMACOLOGIC INHIBITION OF THE EPIGENETIC MODIFIERS HISTONE DEACETYLASES (HDACS). TO ADDRESS THIS AIM, CHRONIC PANCREATITIS WAS INDUCED IN C57BL/6 MICE WITH SERIAL INJECTIONS OF CERULEIN, AND THE SELECTIVE CLASS 1 HDAC INHIBITOR MS-275 WAS ADMINISTERED IN VIVO IN A PREVENTIVE AND THERAPEUTIC MANNER. BOTH MS-275 REGIMENS POTENTLY REDUCED DEPOSITION OF EXTRACELLULAR MATRIX AND DEVELOPMENT OF FIBROSIS IN THE PANCREAS AFTER 4 WEEKS OF CHRONIC PANCREATITIS. REDUCED PANCREATIC FIBROSIS WAS CONCOMITANT WITH LOWER EXPRESSION OF PANCREATIC TGFBETA AND CONSEQUENT REDUCED PSC ACTIVATION. IN SEARCH OF THE CELL TYPES TARGETED BY THE INHIBITOR, WE FOUND THAT MS-275 TREATMENT ABROGATED THE EXPRESSION OF TGFBETA IN ACINAR CELLS STIMULATED BY CERULEIN TREATMENT. OUR STUDY DEMONSTRATES THAT MS-275 IS AN EFFECTIVE ANTIFIBROTIC AGENT IN THE CONTEXT OF EXPERIMENTAL CHRONIC PANCREATITIS AND THUS MAY CONSTITUTE A VALID THERAPEUTIC INTERVENTION FOR THIS SEVERE DISEASE. 2018