1 3677 127 INFLAMMATION AND PYROPTOSIS MEDIATE MUSCLE EXPANSION IN AN INTERLEUKIN-1BETA (IL-1BETA)-DEPENDENT MANNER. MUSCLE INFLAMMATION IS OFTEN ASSOCIATED WITH ITS EXPANSION. BLADDER SMOOTH MUSCLE INFLAMMATION-INDUCED CELL DEATH IS ACCOMPANIED BY HYPERPLASIA AND HYPERTROPHY AS THE PRIMARY CAUSE FOR POOR BLADDER FUNCTION. IN MICE, DNA DAMAGE INITIATED BY CHEMOTHERAPEUTIC DRUG CYCLOPHOSPHAMIDE ACTIVATED CASPASE 1 THROUGH THE FORMATION OF THE NLRP3 COMPLEX RESULTING IN DETRUSOR HYPERPLASIA. A CYCLOPHOSPHAMIDE METABOLITE, ACROLEIN, CAUSED GLOBAL DNA METHYLATION AND ACCUMULATION OF DNA DAMAGE IN A MOUSE MODEL OF BLADDER INFLAMMATION AND IN CULTURED BLADDER MUSCLE CELLS. IN CORRELATION, GLOBAL DNA METHYLATION AND NLRP3 EXPRESSION WAS UP-REGULATED IN HUMAN CHRONIC BLADDER INFLAMMATORY TISSUES. THE EPIGENETIC SILENCING OF DNA DAMAGE REPAIR GENE, OGG1, COULD BE REVERSED BY THE USE OF DEMETHYLATING AGENTS. IN MICE, DEMETHYLATING AGENTS REVERSED CYCLOPHOSPHAMIDE-INDUCED BLADDER INFLAMMATION AND DETRUSOR EXPANSION. THE TRANSGENIC KNOCK-OUT OF OGG1 IN AS FEW AS 10% OF THE DETRUSOR CELLS TRIPLED THE PROLIFERATION OF THE REMAINING WILD TYPE COUNTERPARTS IN AN IN VITRO CO-CULTURE TITRATION EXPERIMENT. ANTAGONIZING IL-1BETA WITH ANAKINRA, A RHEUMATOID ARTHRITIS THERAPEUTIC, PREVENTED DETRUSOR PROLIFERATION IN CONDITIONED MEDIA EXPERIMENTS AS WELL AS IN A MOUSE MODEL OF BLADDER INFLAMMATION. RADIATION TREATMENT VALIDATED THE ROLE OF DNA DAMAGE IN THE NLRP3-ASSOCIATED CASPASE 1-MEDIATED IL-1BETA SECRETORY PHENOTYPE. A PROTEIN ARRAY ANALYSIS IDENTIFIED IGF1 TO BE DOWNSTREAM OF IL-1BETA SIGNALING. IL-1BETA-INDUCED DETRUSOR PROLIFERATION AND HYPERTROPHY COULD BE REVERSED WITH THE USE OF ANAKINRA AS WELL AS AN IGF1 NEUTRALIZING ANTIBODY. IL-1BETA ANTAGONISTS IN CURRENT CLINICAL PRACTICE CAN EXPLOIT THE REVEALED MECHANISM FOR DNA DAMAGE-MEDIATED MUSCULAR EXPANSION. 2015 2 64 31 A HIGH-THROUGHPUT SCREENING ASSAY FOR SILENCING ESTABLISHED HIV-1 MACROPHAGE INFECTION IDENTIFIES NUCLEOSIDE ANALOGS THAT PERTURB H3K9ME3 ON PROVIRAL GENOMES. HIV-INFECTED MACROPHAGES ARE LONG-LIVED CELLS THAT REPRESENT A BARRIER TO FUNCTIONAL CURE. ADDITIONALLY, LOW-LEVEL VIRAL EXPRESSION BY CENTRAL NERVOUS SYSTEM (CNS) MACROPHAGES CONTRIBUTES TO NEUROCOGNITIVE DEFICITS THAT DEVELOP DESPITE ANTIRETROVIRAL THERAPY (ART). WE RECENTLY IDENTIFIED H3K9ME3 AS AN ATYPICAL EPIGENETIC MARK ASSOCIATED WITH CHRONIC HIV INFECTION IN MACROPHAGES. THUS, STRATEGIES ARE NEEDED TO SUPPRESS HIV-1 EXPRESSION IN MACROPHAGES, BUT THE UNIQUE MYELOID ENVIRONMENT AND THE RESPONSIBLE MACROPHAGE/CNS-TROPIC STRAINS REQUIRE CELL/STRAIN-SPECIFIC APPROACHES. HERE, WE GENERATED AN HIV-1 REPORTER VIRUS FROM A CNS-DERIVED STRAIN WITH INTACT AUXILIARY GENES EXPRESSING DESTABILIZED LUCIFERASE. WE EMPLOYED THIS REPORTER VIRUS IN POLYCLONAL INFECTION OF PRIMARY HUMAN MONOCYTE-DERIVED MACROPHAGES (MDM) FOR A HIGH-THROUGHPUT SCREEN (HTS) TO IDENTIFY COMPOUNDS THAT SUPPRESS VIRUS EXPRESSION FROM ESTABLISHED MACROPHAGE INFECTION. SCREENING ~6,000 KNOWN DRUGS AND COMPOUNDS YIELDED 214 HITS. A SECONDARY SCREEN WITH 10-DOSE TITRATION IDENTIFIED 24 MEETING CRITERIA FOR HIV-SELECTIVE ACTIVITY. USING THREE REPLICATION-COMPETENT CNS-DERIVED MACROPHAGE-TROPIC HIV-1 ISOLATES AND VIRAL GENE EXPRESSION READOUT IN MDM, WE CONFIRMED THE EFFECT OF THREE PURINE ANALOGS, NELARABINE, FLUDARABINE, AND ENTECAVIR, SHOWING THE SUPPRESSION OF HIV-1 EXPRESSION FROM ESTABLISHED MACROPHAGE INFECTION. NELARABINE INHIBITED THE FORMATION OF H3K9ME3 ON HIV GENOMES IN MACROPHAGES. THUS, THIS NOVEL HTS ASSAY CAN IDENTIFY SUPPRESSORS OF HIV-1 TRANSCRIPTION IN ESTABLISHED MACROPHAGE INFECTION, SUCH AS NUCLEOSIDE ANALOGS AND HDAC INHIBITORS, WHICH MAY BE LINKED TO H3K9ME3 MODIFICATION. THIS SCREEN MAY BE USEFUL TO IDENTIFY NEW METABOLIC AND EPIGENETIC AGENTS THAT AMELIORATE HIV-DRIVEN NEUROINFLAMMATION IN PEOPLE ON ART OR PREVENT VIRAL RECRUDESCENCE FROM MACROPHAGE RESERVOIRS IN STRATEGIES TO ACHIEVE ART-FREE REMISSION. IMPORTANCE MACROPHAGES INFECTED BY HIV-1 ARE A LONG-LIVED RESERVOIR AND A BARRIER IN CURRENT EFFORTS TO ACHIEVE HIV CURE AND ALSO CONTRIBUTE TO NEUROCOGNITIVE COMPLICATIONS IN PEOPLE DESPITE ANTIRETROVIRAL THERAPY (ART). SILENCING HIV EXPRESSION IN THESE CELLS WOULD BE OF GREAT VALUE, BUT THE REGULATION OF HIV-1 IN MACROPHAGES DIFFERS FROM T CELLS. WE DEVELOPED A NOVEL HIGH-THROUGHPUT SCREEN FOR COMPOUNDS THAT CAN SILENCE ESTABLISHED INFECTION OF PRIMARY MACROPHAGES, AND IDENTIFIED AGENTS THAT DOWNREGULATE VIRUS EXPRESSION AND ALTER PROVIRUS EPIGENETIC PROFILES. THE SIGNIFICANCE OF THIS ASSAY IS THE POTENTIAL TO IDENTIFY NEW DRUGS THAT ACT IN THE UNIQUE MACROPHAGE ENVIRONMENT ON RELEVANT VIRAL STRAINS, WHICH MAY CONTRIBUTE TO ADJUNCTIVE TREATMENT FOR HIV-ASSOCIATED NEUROCOGNITIVE DISORDERS AND/OR PREVENT VIRAL REBOUND IN EFFORTS TO ACHIEVE ART-FREE REMISSION OR CURE. 2023 3 6523 28 TRANSCRIPTIONAL AND EPIGENETIC REGULATORY MECHANISMS AFFECTING HTLV-1 PROVIRUS. HUMAN T-CELL LEUKEMIA VIRUS TYPE 1 (HTLV-1) IS A RETROVIRUS ASSOCIATED WITH HUMAN DISEASES, SUCH AS ADULT T-CELL LEUKEMIA (ATL) AND HTLV-1-ASSOCIATED MYELOPATHY/TROPIC SPASTIC PARAPARESIS (HAM/TSP). AS A RETROVIRUS, ITS LIFE CYCLE INCLUDES A STEP WHERE HTLV-1 IS INTEGRATED INTO THE HOST GENOMIC DNA AND FORMS PROVIRAL DNA. IN THE CHRONIC PHASE OF THE INFECTION, HTLV?1 IS KNOWN TO PROLIFERATE AS A PROVIRUS VIA THE MITOTIC DIVISION OF THE INFECTED HOST CELLS. THERE ARE GENERALLY TENS OF THOUSANDS OF INFECTED CLONES WITHIN AN INFECTED INDIVIDUAL. THEY EXIST NOT ONLY IN PERIPHERAL BLOOD, BUT ALSO IN VARIOUS LYMPHOID ORGANS. VIRAL PROTEINS ENCODED IN HTLV-1 GENOME PLAY A ROLE IN THE PROLIFERATION AND SURVIVAL OF THE INFECTED CELLS. AS IS THE CASE WITH OTHER CHRONIC VIRAL INFECTIONS, HTLV-1 GENE EXPRESSION INDUCES THE ACTIVATION OF THE HOST IMMUNITY AGAINST THE VIRUS. THUS, THE TRANSCRIPTION FROM HTLV-1 PROVIRUS NEEDS TO BE CONTROLLED IN ORDER TO EVADE THE HOST IMMUNE SURVEILLANCE. THERE SHOULD BE A DYNAMIC AND COMPLEX REGULATION IN VIVO, WHERE AN EQUILIBRIUM BETWEEN VIRAL ANTIGEN EXPRESSION AND HOST IMMUNE SURVEILLANCE IS ACHIEVED. THE MECHANISMS REGULATING VIRAL GENE EXPRESSION FROM THE PROVIRUS ARE A KEY TO UNDERSTANDING THE PERSISTENT/LATENT INFECTION WITH HTLV-1 AND ITS PATHOGENESIS. IN THIS ARTICLE, WE WOULD LIKE TO REVIEW OUR CURRENT UNDERSTANDING ON THIS TOPIC. 2016 4 2143 25 EPIGENETIC LANDSCAPE OF HIV-1 INFECTION IN PRIMARY HUMAN MACROPHAGE. HUMAN IMMUNODEFICIENCY VIRUS (HIV)-INFECTED MACROPHAGES ARE LONG-LIVED CELLS THAT SUSTAIN PERSISTENT VIRUS EXPRESSION, WHICH IS BOTH A BARRIER TO VIRAL ERADICATION AND CONTRIBUTOR TO NEUROLOGICAL COMPLICATIONS IN PATIENTS DESPITE ANTIRETROVIRAL THERAPY (ART). TO BETTER UNDERSTAND THE REGULATION OF HIV-1 IN MACROPHAGES, WE COMPARED HIV-INFECTED PRIMARY HUMAN MONOCYTE-DERIVED MACROPHAGES (MDM) TO ACUTELY INFECTED PRIMARY CD4 T CELLS AND JURKAT CELLS LATENTLY INFECTED WITH HIV (JLAT 8.4). HIV GENOMES IN MDM WERE ACTIVELY TRANSCRIBED DESPITE ENRICHMENT WITH HETEROCHROMATIN-ASSOCIATED H3K9ME3 ACROSS THE COMPLETE HIV GENOME IN COMBINATION WITH ELEVATED ACTIVATION MARKS OF H3K9AC AND H3K27AC AT THE LONG TERMINAL REPEAT (LTR). MACROPHAGE PATTERNS CONTRASTED WITH JLAT CELLS, WHICH SHOWED CONVENTIONAL BIVALENT H3K4ME3/H3K27ME3, AND ACUTELY INFECTED CD4 T CELLS, WHICH SHOWED AN INTERMEDIATE EPIGENOTYPE. 5'-METHYLCYTOSINE (5MC) WAS ENRICHED ACROSS THE HIV GENOME IN LATENTLY INFECTED JLAT CELLS, WHILE 5'-HYDROXYMETHYLCYTOSINE (5HMC) WAS ENRICHED IN CD4 CELLS AND MDMS. HIV INFECTION INDUCED MULTINUCLEATION OF MDMS ALONG WITH DNA DAMAGE-ASSOCIATED P53 PHOSPHORYLATION, AS WELL AS LOSS OF TET2 AND THE NUCLEAR REDISTRIBUTION OF 5-HYDOXYMETHYLATION. TAKEN TOGETHER, OUR FINDINGS SUGGEST THAT HIV INDUCES A UNIQUE MACROPHAGE NUCLEAR AND TRANSCRIPTIONAL PROFILE, AND VIRAL GENOMES ARE MAINTAINED IN A NONCANONICAL BIVALENT EPIGENETIC STATE. IMPORTANCE MACROPHAGES SERVE AS A RESERVOIR FOR LONG-TERM PERSISTENCE AND CHRONIC PRODUCTION OF HIV. WE FOUND AN ATYPICAL EPIGENETIC CONTROL OF HIV IN MACROPHAGES MARKED BY HETEROCHROMATIC H3K9ME3 DESPITE ACTIVE VIRAL TRANSCRIPTION. HIV INFECTION INDUCED CHANGES IN MACROPHAGE NUCLEAR MORPHOLOGY AND EPIGENETIC REGULATORY FACTORS. THESE FINDINGS MAY IDENTIFY NEW MECHANISMS TO CONTROL CHRONIC HIV EXPRESSION IN INFECTED MACROPHAGES. 2022 5 6688 21 VALPROATE SYNERGIZES WITH PURINE NUCLEOSIDE ANALOGUES TO INDUCE APOPTOSIS OF B-CHRONIC LYMPHOCYTIC LEUKAEMIA CELLS. RESISTANCE TO CHEMOTHERAPY AND DRUG TOXICITY ARE TWO MAJOR CONCERNS OF CHRONIC LYMPHOCYTIC LEUKAEMIA (B-CLL) TREATMENT BY PURINE NUCLEOSIDE ANALOGUES (PNA, I.E. FLUDARABINE AND CLADRIBINE). WE HYPOTHESIZED THAT TARGETING EPIGENETIC CHANGES MIGHT ADDRESS THESE ISSUES AND EVALUATED THE EFFECT OF THE HISTONE DEACETYLASE INHIBITOR VALPROATE (VPA) AT A CLINICALLY RELEVANT CONCENTRATION. VPA ACTED IN A HIGHLY SYNERGISTIC/ADDITIVE MANNER WITH FLUDARABINE AND CLADRIBINE TO INDUCE APOPTOSIS OF B-CLL CELLS. IMPORTANTLY, VPA ALSO RESTORED SENSITIVITY TO FLUDARABINE IN B CELLS FROM POOR PROGNOSIS CLL PATIENTS WHO BECAME RESISTANT TO CHEMOTHERAPY. MECHANISM OF APOPTOSIS INDUCED BY VPA ALONE OR COMBINED WITH FLUDARABINE OR TO CLADRIBINE WAS CASPASE-DEPENDENT AND INVOLVED THE EXTRINSIC PATHWAY. VPA, BUT NEITHER FLUDARABINE NOR CLADRIBINE, ENHANCED THE PRODUCTION OF REACTIVE OXYGEN SPECIES (ROS) AND INHIBITION OF ROS WITH N-ACETYLCYSTEINE DECREASES APOPTOSIS OF CLL CELLS. VPA STIMULATES HYPERPHOSPHORYLATION OF P42/P44 ERK, CYTOCHROME C RELEASE AND OVEREXPRESSION OF BAX AND FAS. TOGETHER, OUR DATA INDICATE THAT VPA MAY AMELIORATE THE OUTCOME OF PNA-BASED THERAPEUTIC PROTOCOLS AND PROVIDE A POTENTIAL ALTERNATIVE TREATMENT IN BOTH THE RELAPSED AND FRONT-LINE RESISTANT PATIENTS AND IN PATIENTS WITH HIGH RISK FEATURES. 2009 6 3380 31 HIV-1 INFECTION OF GENETICALLY ENGINEERED IPSC-DERIVED CENTRAL NERVOUS SYSTEM-ENGRAFTED MICROGLIA IN A HUMANIZED MOUSE MODEL. THE CENTRAL NERVOUS SYSTEM (CNS) IS A MAJOR HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 RESERVOIR. MICROGLIA ARE THE PRIMARY TARGET CELL OF HIV-1 INFECTION IN THE CNS. CURRENT MODELS HAVE NOT ALLOWED THE PRECISE MOLECULAR PATHWAYS OF ACUTE AND CHRONIC CNS MICROGLIAL INFECTION TO BE TESTED WITH IN VIVO GENETIC METHODS. HERE, WE DESCRIBE A NOVEL HUMANIZED MOUSE MODEL UTILIZING HUMAN-INDUCED PLURIPOTENT STEM CELL-DERIVED MICROGLIA TO XENOGRAFT INTO MURINE HOSTS. THESE MICE ARE ADDITIONALLY ENGRAFTED WITH HUMAN PERIPHERAL BLOOD MONONUCLEAR CELLS THAT SERVED AS A MEDIUM TO ESTABLISH A PERIPHERAL INFECTION THAT THEN SPREAD TO THE CNS MICROGLIA XENOGRAFT, MODELING A TRANS-BLOOD-BRAIN BARRIER ROUTE OF ACUTE CNS HIV-1 INFECTION WITH HUMAN TARGET CELLS. THE APPROACH IS COMPATIBLE WITH IPSC GENETIC ENGINEERING, INCLUDING INSERTING TARGETED TRANSGENIC REPORTER CASSETTES TO TRACK THE XENOGRAFTED HUMAN CELLS, ENABLING THE TESTING OF NOVEL TREATMENT AND VIRAL TRACKING STRATEGIES IN A COMPARATIVELY SIMPLE AND COST-EFFECTIVE WAY VIVO MODEL FOR NEUROHIV. IMPORTANCE: OUR MOUSE MODEL IS A POWERFUL TOOL FOR INVESTIGATING THE GENETIC MECHANISMS GOVERNING CNS HIV-1 INFECTION AND LATENCY IN THE CNS AT A SINGLE-CELL LEVEL. A MAJOR ADVANTAGE OF OUR MODEL IS THAT IT USES IPSC-DERIVED MICROGLIA, WHICH ENABLES HUMAN GENETICS, INCLUDING GENE FUNCTION AND THERAPEUTIC GENE MANIPULATION, TO BE EXPLORED IN VIVO , WHICH IS MORE CHALLENGING TO STUDY WITH CURRENT HEMATOPOIETIC STEM CELL-BASED MODELS FOR NEUROHIV. OUR TRANSGENIC TRACING OF XENOGRAFTED HUMAN CELLS WILL PROVIDE A QUANTITATIVE MEDIUM TO DEVELOP NEW MOLECULAR AND EPIGENETIC STRATEGIES FOR REDUCING THE HIV-1 LATENT RESERVOIR AND TO TEST THE IMPACT OF THERAPEUTIC INFLAMMATION-TARGETING DRUG INTERVENTIONS ON CNS HIV-1 LATENCY. 2023 7 3877 31 KDM6A PROMOTES IMATINIB RESISTANCE THROUGH YY1-MEDIATED TRANSCRIPTIONAL UPREGULATION OF TRKA INDEPENDENTLY OF ITS DEMETHYLASE ACTIVITY IN CHRONIC MYELOGENOUS LEUKEMIA. RATIONALE: DESPITE LANDMARK THERAPY OF CHRONIC MYELOGENOUS LEUKEMIA (CML) WITH TYROSINE KINASE INHIBITORS (TKIS), DRUG RESISTANCE REMAINS PROBLEMATIC. CANCER PATHOGENESIS INVOLVES EPIGENETIC DYSREGULATION AND IN PARTICULAR, HISTONE LYSINE DEMETHYLASES (KDMS) HAVE BEEN IMPLICATED IN TKI RESISTANCE. WE SOUGHT TO IDENTIFY KDMS WITH ALTERED EXPRESSION IN CML AND DEFINE THEIR CONTRIBUTION TO IMATINIB RESISTANCE. METHODS: BIOINFORMATICS SCREENING COMPARED KDM EXPRESSION IN CML VERSUS NORMAL BONE MARROW WITH SHRNA KNOCKDOWN AND FLOW CYTOMETRY USED TO MEASURE EFFECTS ON IMATINIB-INDUCED APOPTOSIS IN K562 CELLS. TRANSCRIPTOMIC ANALYSES WERE PERFORMED AGAINST KDM6A CRISPR KNOCKOUT/SHRNA KNOCKDOWN K562 CELLS ALONG WITH GENE RESCUE EXPERIMENTS USING WILDTYPE AND MUTANT DEMETHYLASE-DEAD KDM6A CONSTRUCTS. CO-IMMUNOPRECIPITATION, LUCIFERASE REPORTER AND CHIP WERE EMPLOYED TO ELUCIDATE MECHANISMS OF KDM6A-DEPENDENT RESISTANCE. RESULTS: AMONGST FIVE KDMS UPREGULATED IN CML, ONLY KDM6A DEPLETION SENSITIZED CML CELLS TO IMATINIB-INDUCED APOPTOSIS. RE-INTRODUCTION OF DEMETHYLASE-DEAD KDM6A AS WELL AS WILD-TYPE KDM6A RESTORED IMATINIB RESISTANCE. RNA-SEQ IDENTIFIED NTRK1 GENE DOWNREGULATION AFTER DEPLETION OF KDM6A. MOREOVER, NTRK1 EXPRESSION POSITIVELY CORRELATED WITH KDM6A IN A SUBSET OF CLINICAL CML SAMPLES AND KDM6A KNOCKDOWN IN FRESH CML ISOLATES DECREASED NTRK1 ENCODED PROTEIN (TRKA) EXPRESSION. MECHANISTICALLY, KDM6A WAS RECRUITED TO THE NTRK1 PROMOTER BY THE TRANSCRIPTION FACTOR YY1 WITH SUBSEQUENT TRKA UPREGULATION ACTIVATING DOWN-STREAM SURVIVAL PATHWAYS TO INVOKE IMATINIB RESISTANCE. CONCLUSION: CONTRARY TO ITS REPORTED ROLE AS A TUMOR SUPPRESSOR AND INDEPENDENT OF ITS DEMETHYLASE FUNCTION, KDM6A PROMOTES IMATINIB-RESISTANCE IN CML CELLS. THE IDENTIFICATION OF THE KDM6A/YY1/TRKA AXIS AS A NOVEL IMATINIB-RESISTANCE MECHANISM REPRESENTS AN UNEXPLORED AVENUE TO OVERCOME TKI RESISTANCE IN CML. 2021 8 1853 31 ELEVATED METHIONINE FLUX DRIVES PYROPTOSIS EVASION IN PERSISTER CANCER CELLS. INDUCTION OF CELL DEATH REPRESENTS A PRIMARY GOAL OF MOST ANTICANCER TREATMENTS. DESPITE THE EFFICACY OF SUCH APPROACHES, A SMALL POPULATION OF "PERSISTERS" DEVELOP EVASION STRATEGIES TO THERAPY-INDUCED CELL DEATH. WHILE PREVIOUS STUDIES HAVE IDENTIFIED MECHANISMS OF RESISTANCE TO APOPTOSIS, THE MECHANISMS BY WHICH PERSISTERS DAMPEN OTHER FORMS OF CELL DEATH, SUCH AS PYROPTOSIS, REMAIN TO BE ELUCIDATED. PYROPTOSIS IS A FORM OF INFLAMMATORY CELL DEATH THAT INVOLVES FORMATION OF MEMBRANE PORES, ION GRADIENT IMBALANCE, WATER INFLOW, AND MEMBRANE RUPTURE. HEREIN, WE INVESTIGATE MECHANISMS BY WHICH CANCER PERSISTERS RESIST PYROPTOSIS, SURVIVE, THEN PROLIFERATE IN THE PRESENCE OF TYROSINE KINASE INHIBITORS (TKI). LUNG, PROSTATE, AND ESOPHAGEAL CANCER PERSISTER CELLS REMAINING AFTER TREATMENTS EXHIBITED SEVERAL HALLMARKS INDICATIVE OF PYROPTOSIS RESISTANCE. THE INFLAMMATORY ATTRIBUTES OF PERSISTERS INCLUDED CHRONIC ACTIVATION OF INFLAMMASOME, STING, AND TYPE I INTERFERONS. COMPREHENSIVE METABOLOMIC CHARACTERIZATION UNCOVERED THAT TKI-INDUCED PYROPTOTIC PERSISTERS DISPLAY HIGH METHIONINE CONSUMPTION AND EXCESSIVE TAURINE PRODUCTION. ELEVATED METHIONINE FLUX OR EXOGENOUS TAURINE PRESERVED PLASMA MEMBRANE INTEGRITY VIA OSMOLYTE-MEDIATED EFFECTS. INCREASED DEPENDENCY ON METHIONINE FLUX DECREASED THE LEVEL OF ONE CARBON METABOLISM INTERMEDIATE S-(5'-ADENOSYL)-L-HOMOCYSTEINE, A DETERMINANT OF CELL METHYLATION CAPACITY. THE CONSEQUENT INCREASE IN METHYLATION POTENTIAL INDUCED DNA HYPERMETHYLATION OF GENES REGULATING METAL ION BALANCE AND INTRINSIC IMMUNE RESPONSE. THIS ENABLED THWARTING TKI RESISTANCE BY USING THE HYPOMETHYLATING AGENT DECITABINE. IN SUMMARY, THE EVOLUTION OF RESISTANCE TO PYROPTOSIS CAN OCCUR VIA A STEPWISE PROCESS OF PHYSICAL ACCLIMATION AND EPIGENETIC CHANGES WITHOUT EXISTING OR RECURRENT MUTATIONS. SIGNIFICANCE: METHIONINE ENABLES CANCER CELLS TO PERSIST BY EVADING PYROPTOTIC OSMOTIC LYSIS, WHICH LEADS TO GENOME-WIDE HYPERMETHYLATION THAT ALLOWS PERSISTERS TO GAIN PROLIFERATIVE ADVANTAGES. 2023 9 3340 32 HISTONE DEACETYLASE MEDIATED TRANSCRIPTIONAL ACTIVATION REDUCES PROVIRAL LOADS IN HTLV-1 ASSOCIATED MYELOPATHY/TROPICAL SPASTIC PARAPARESIS PATIENTS. EPIGENETIC MODIFICATIONS OF CHROMATIN MAY PLAY A ROLE IN MAINTAINING VIRAL LATENCY AND THUS PERSISTENCE OF THE HUMAN T-LYMPHOTROPIC VIRUS TYPE 1 (HTLV-1), WHICH IS RESPONSIBLE FOR HTLV-ASSOCIATED MYELOPATHY/TROPICAL SPASTIC PARAPARESIS (HAM/TSP). A MAJOR DETERMINANT OF DISEASE PROGRESSION IS INCREASED PERIPHERAL BLOOD PROVIRAL LOAD (PVL), POSSIBLY VIA THE ACCUMULATION OF INFECTED CELLS IN THE CENTRAL NERVOUS SYSTEM (CNS) CREATING A DAMAGING INFLAMMATORY RESPONSE. CURRENT THERAPEUTIC APPROACHES THAT FOCUS ON REDUCING EITHER CELL PROLIFERATION, VIRAL REPLICATION, OR TISSUE INVASION ARE STILL UNSATISFACTORY. CONTRASTING WITH THESE INHIBITORY STRATEGIES, WE EVALUATED THE EFFICACY OF A NOVEL APPROACH AIMED, PARADOXICALLY, AT ACTIVATING VIRAL GENE EXPRESSION TO EXPOSE VIRUS-POSITIVE CELLS TO THE HOST IMMUNE RESPONSE. WE USED VALPROATE (VPA), A HISTONE DEACETYLASE INHIBITOR THAT HAS BEEN USED FOR DECADES AS A CHRONIC, SAFE TREATMENT FOR EPILEPTIC DISORDERS. BASED ON IN VITRO AND IN VIVO DATA, WE PROVIDE EVIDENCE THAT TRANSIENT ACTIVATION OF THE LATENT VIRAL RESERVOIR CAUSES ITS COLLAPSE, A PROCESS THAT MAY ALLEVIATE THE CONDITION OF HAM/TSP. THIS REPRESENTS THE FIRST SUCH APPROACH TO TREATING HAM/TSP, USING GENE ACTIVATION THERAPY TO TILT THE HOST-PATHOGEN BALANCE IN FAVOR OF AN EXISTING ANTIVIRAL RESPONSE. THIS TRIAL IS REGISTERED AT HTTP://CLINICALTRIALS.GOV/AS NO. NCT00519181. 2007 10 2115 15 EPIGENETIC HETEROGENEITY IN HIV-1 LATENCY ESTABLISHMENT. DESPITE PROLONGED ANTIRETROVIRAL THERAPY, HIV-1 PERSISTS AS TRANSCRIPTIONALLY INACTIVE PROVIRUSES. THE HIV-1 LATENCY REMAINS A PRINCIPAL OBSTACLE IN CURING AIDS. IT IS IMPORTANT TO UNDERSTAND MECHANISMS BY WHICH HIV-1 LATENCY IS ESTABLISHED TO MAKE THE LATENT RESERVOIR SMALLER. WE PRESENT A MOLECULAR CHARACTERIZATION OF DISTINCT POPULATIONS AT AN EARLY PHASE OF INFECTION. WE DEVELOPED AN ORIGINAL DUAL-COLOR REPORTER VIRUS TO MONITOR LTR KINETICS FROM ESTABLISHMENT TO MAINTENANCE STAGE. WE FOUND THAT THERE ARE TWO WAYS OF LATENCY ESTABLISHMENT I.E., BY IMMEDIATE SILENCING AND SLOW INACTIVATION FROM ACTIVE INFECTION. HISTONE COVALENT MODIFICATIONS, PARTICULARLY POLYCOMB REPRESSIVE COMPLEX 2 (PRC2)-MEDIATED H3K27 TRIMETHYLATION, APPEARED TO DOMINATE VIRAL TRANSCRIPTION AT THE EARLY PHASE. PRC2 ALSO CONTRIBUTES TO TIME-DEPENDENT LTR DORMANCY IN THE CHRONIC PHASE OF THE INFECTION. SIGNIFICANT DIFFERENCES IN SENSITIVITY AGAINST SEVERAL STIMULI WERE OBSERVED BETWEEN THESE TWO DISTINCT POPULATIONS. THESE RESULTS WILL EXPAND OUR UNDERSTANDING OF HETEROGENEOUS ESTABLISHMENT OF HIV-1 LATENCY POPULATIONS. 2015 11 2392 25 EPIGENETIC REPRESSION OF INTERLEUKIN 2 EXPRESSION IN SENESCENT CD4+ T CELLS DURING CHRONIC HIV TYPE 1 INFECTION. THE MOLECULAR MECHANISMS FOR IL2 GENE-SPECIFIC DYSREGULATION DURING CHRONIC HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 (HIV-1) INFECTION ARE UNKNOWN. HERE, WE INVESTIGATED THE ROLE OF DNA METHYLATION IN SUPPRESSING INTERLEUKIN 2 (IL-2) EXPRESSION IN MEMORY CD4(+) T CELLS DURING CHRONIC HIV-1 INFECTION. WE OBSERVED THAT CPG SITES IN THE IL2 PROMOTER OF CD4(+) T CELLS WERE FULLY METHYLATED IN NAIVE CD4(+) T CELLS AND SIGNIFICANTLY DEMETHYLATED IN THE MEMORY POPULATIONS. INTERESTINGLY, WE FOUND THAT THE MEMORY CELLS THAT HAD A TERMINALLY DIFFERENTIATED PHENOTYPE AND EXPRESSED CD57 HAD INCREASED IL2 PROMOTER METHYLATION RELATIVE TO LESS DIFFERENTIATED MEMORY CELLS IN HEALTHY INDIVIDUALS. IMPORTANTLY, EARLY EFFECTOR MEMORY SUBSETS FROM HIV-1-INFECTED SUBJECTS EXPRESSED HIGH LEVELS OF CD57 AND WERE HIGHLY METHYLATED AT THE IL2 LOCUS. FURTHERMORE, THE INCREASED CD57 EXPRESSION ON MEMORY CD4(+) T CELLS WAS INVERSELY CORRELATED WITH IL-2 PRODUCTION. THESE DATA SUGGEST THAT DNA METHYLATION AT THE IL2 LOCUS IN CD4(+) T CELLS IS COUPLED TO IMMUNOSENESCENCE AND PLAYS A CRITICAL ROLE IN THE BROAD DYSFUNCTION THAT OCCURS IN POLYCLONAL T CELLS DURING HIV-1 INFECTION. 2015 12 4849 25 OPIOID-MEDIATED HIV-1 IMMUNOPATHOGENESIS. DESPITE THE ABILITY OF COMBINATION ANTIRETROVIRAL THERAPY TO DRAMATICALLY SUPPRESS VIREMIA, THE BRAIN CONTINUES TO BE A RESERVOIR OF HIV-1 LOW-LEVEL REPLICATION. ADDING FURTHER COMPLEXITY TO THIS IS THE COMORBIDITY OF DRUG ABUSE WITH HIV-1 ASSOCIATED NEUROCOGNITIVE DISORDERS AND NEUROHIV. AMONG SEVERAL ABUSED DRUGS, THE USE OF OPIATES IS HIGHLY PREVALENT IN HIV-1 INFECTED INDIVIDUALS, BOTH AS AN ABUSED DRUG AS WELL AS FOR PAIN MANAGEMENT. OPIOIDS AND THEIR RECEPTORS HAVE ATTAINED NOTABLE ATTENTION OWING TO THEIR ABILITY TO MODULATE IMMUNE FUNCTIONS, IN TURN, IMPACTING DISEASE PROGRESSION. VARIOUS CELL CULTURE, ANIMAL AND HUMAN STUDIES HAVE IMPLICATED THE ROLE OF OPIOIDS AND THEIR RECEPTORS IN MODULATING VIRAL REPLICATION AND VIRUS-MEDIATED PATHOLOGY BOTH POSITIVELY AND NEGATIVELY. FURTHER, THE COMBINATORIAL EFFECTS OF HIV-1/HIV-1 PROTEINS AND MORPHINE HAVE DEMONSTRATED ACTIVATION OF INFLAMMATORY SIGNALING IN THE HOST SYSTEM. HEREIN, WE SUMMARIZED THE CURRENT KNOWLEDGE ON THE ROLE OF OPIOIDS ON PERIPHERAL IMMUNOPATHOGENESIS, VIRAL IMMUNOPATHOGENESIS, EPIGENETIC PROFILES OF THE HOST AND VIRAL GENOME, NEUROPATHOGENESIS OF SIV/SHIV-INFECTED NON-HUMAN PRIMATES, BLOOD-BRAIN-BARRIER, HIV-1 VIRAL LATENCY, AND VIRAL REBOUND. OVERALL, THIS REVIEW PROVIDES RECENT INSIGHTS INTO THE ROLE OF OPIOIDS IN HIV-1 IMMUNOPATHOGENESIS. GRAPHICAL ABSTRACT. 2020 13 2326 29 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 14 5908 29 TARGETED DE-METHYLATION OF THE FOXP3-TSDR IS SUFFICIENT TO INDUCE PHYSIOLOGICAL FOXP3 EXPRESSION BUT NOT A FUNCTIONAL TREG PHENOTYPE. CD4+ REGULATORY T CELLS (TREGS) ARE KEY MEDIATORS OF IMMUNOLOGICAL TOLERANCE AND PROMISING EFFECTOR CELLS FOR IMMUNO-SUPPRESSIVE ADOPTIVE CELLULAR THERAPY TO FIGHT AUTOIMMUNITY AND CHRONIC INFLAMMATION. THEIR FUNCTIONAL STABILITY IS CRITICAL FOR THEIR CLINICAL UTILITY AND HAS BEEN CORRELATED TO THE DEMETHYLATED STATE OF THE TSDR/CNS2 ENHANCER ELEMENT IN THE TREG LINEAGE TRANSCRIPTION FACTOR FOXP3. HOWEVER, PROOF FOR A CAUSAL CONTRIBUTION OF THE TSDR DE-METHYLATION TO FOXP3 STABILITY AND TREG INDUCTION IS SO FAR LACKING. WE HERE ESTABLISHED A POWERFUL TRANSIENT-TRANSFECTION CRISPR-CAS9-BASED EPIGENETIC EDITING METHOD FOR THE SELECTIVE DE-METHYLATION OF THE TSDR WITHIN THE ENDOGENOUS CHROMATIN ENVIRONMENT OF A LIVING CELL. THE INDUCED DE-METHYLATED STATE WAS STABLE OVER WEEKS IN CLONAL T CELL PROLIFERATION CULTURES EVEN AFTER EXPRESSION OF THE EDITING COMPLEX HAD CEASED. EPIGENETIC EDITING OF THE TSDR RESULTED IN FOXP3 EXPRESSION, EVEN IN ITS PHYSIOLOGICAL ISOFORM DISTRIBUTION, PROVING A CAUSAL ROLE FOR THE DE-METHYLATED TSDR IN FOXP3 REGULATION. HOWEVER, SUCCESSFUL FOXP3 INDUCTION WAS NOT ASSOCIATED WITH A SWITCH TOWARDS A FUNCTIONAL TREG PHENOTYPE, IN CONTRAST TO WHAT HAS BEEN REPORTED FROM FOXP3 OVEREXPRESSION APPROACHES. THUS, TSDR DE-METHYLATION IS REQUIRED, BUT NOT SUFFICIENT FOR A STABLE TREG PHENOTYPE INDUCTION. THEREFORE, TARGETED DEMETHYLATION OF THE TSDR MAY BE A CRITICAL ADDITION TO PUBLISHED IN VITRO TREG INDUCTION PROTOCOLS WHICH SO FAR LACK FOXP3 STABILITY. 2020 15 4304 31 MICRORNA-223 PROTECTS NEURONS FROM DEGENERATION IN EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS. MULTIPLE SCLEROSIS IS A CHRONIC INFLAMMATORY, DEMYELINATING, AND NEURODEGENERATIVE DISEASE AFFECTING THE BRAIN, SPINAL CORD AND OPTIC NERVES. NEURONAL DAMAGE IS TRIGGERED BY VARIOUS HARMFUL FACTORS THAT ENGAGE DIVERSE SIGNALLING CASCADES IN NEURONS; THUS, THERAPEUTIC APPROACHES TO PROTECT NEURONS WILL NEED TO FOCUS ON AGENTS THAT CAN TARGET MULTIPLE BIOLOGICAL PROCESSES. WE HAVE THEREFORE FOCUSED OUR ATTENTION ON MICRORNAS: SMALL NON-CODING RNAS THAT PRIMARILY FUNCTION AS POST-TRANSCRIPTIONAL REGULATORS THAT TARGET MESSENGER RNAS AND REPRESS THEIR TRANSLATION INTO PROTEINS. A SINGLE MICRORNA CAN TARGET MANY FUNCTIONALLY RELATED MESSENGER RNAS MAKING MICRORNAS POWERFUL EPIGENETIC REGULATORS. DYSREGULATION OF MICRORNAS HAS BEEN DESCRIBED IN MANY NEURODEGENERATIVE DISEASES INCLUDING MULTIPLE SCLEROSIS. HERE, WE REPORT THAT TWO MICRORNAS, MIR-223-3P AND MIR-27A-3P, ARE UPREGULATED IN NEURONS IN THE EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS MOUSE MODEL OF CNS INFLAMMATION AND IN GREY MATTER-CONTAINING MULTIPLE SCLEROSIS LESIONS. PRIOR WORK HAS SHOWN PERIPHERAL BLOOD MONONUCLEAR CELL CONDITIONED MEDIA CAUSES SUBLETHAL DEGENERATION OF NEURONS IN CULTURE. WE FIND OVEREXPRESSION OF MIR-27A-3P OR MIR-223-3P PROTECTS DISSOCIATED CORTICAL NEURONS FROM CONDITION MEDIA MEDIATED DEGENERATION. INTRODUCTION OF MIR-223-3P IN VIVO IN MOUSE RETINAL GANGLION CELLS PROTECTS THEIR AXONS FROM DEGENERATION IN EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS. IN SILICO ANALYSIS REVEALED THAT MESSENGER RNAS INVOLVED IN GLUTAMATE RECEPTOR SIGNALLING ARE ENRICHED AS MIR-27A-3P AND MIR-223-3P TARGETS. WE OBSERVE THAT ANTAGONISM OF NMDA AND AMPA TYPE GLUTAMATE RECEPTORS PROTECTS NEURONS FROM CONDITION MEDIA DEPENDENT DEGENERATION. OUR RESULTS SUGGEST THAT MIR-223-3P AND MIR-27A-3P ARE UPREGULATED IN RESPONSE TO INFLAMMATION TO MEDIATE A COMPENSATORY NEUROPROTECTIVE GENE EXPRESSION PROGRAM THAT DESENSITIZES NEURONS TO GLUTAMATE BY TARGETING MESSENGER RNAS INVOLVED IN GLUTAMATE RECEPTOR SIGNALLING. 2019 16 5857 27 SUBSTRATE-SPECIFIC BINDING OF 8-OXOGUANINE DNA GLYCOSYLASE 1 (OGG1) REPROGRAMS MUCOSAL ADAPTATIONS TO CHRONIC AIRWAY INJURY. RECENT ADVANCES HAVE UNCOVERED THE NON-RANDOM DISTRIBUTION OF 7, 8-DIHYDRO-8-OXOGUANINE (8-OXOGUA) INDUCED BY REACTIVE OXYGEN SPECIES, WHICH IS BELIEVED TO HAVE EPIGENETIC EFFECTS. ITS COGNATE REPAIR PROTEIN, 8-OXOGUANINE DNA GLYCOSYLASE 1 (OGG1), READS OXIDATIVE SUBSTRATES AND PARTICIPATES IN TRANSCRIPTIONAL INITIATION. WHEN REDOX SIGNALING IS ACTIVATED IN SMALL AIRWAY EPITHELIAL CELLS, THE DNA REPAIR FUNCTION OF OGG1 IS REPURPOSED TO TRANSMIT ACUTE INFLAMMATORY SIGNALS ACCOMPANIED BY CELL STATE TRANSITIONS AND MODIFICATION OF THE EXTRACELLULAR MATRIX. EPITHELIAL-MESENCHYMAL AND EPITHELIAL-IMMUNE INTERACTIONS ACT COOPERATIVELY TO ESTABLISH A LOCAL NICHE THAT INSTRUCTS THE MUCOSAL IMMUNE LANDSCAPE. IF THE TRANSITIONAL CELL STATE GOVERNED BY OGG1 REMAINS RESPONSIVE TO INFLAMMATORY MEDIATORS INSTEAD OF DIFFERENTIATION, THE COLLATERAL DAMAGE PROVIDES POSITIVE FEEDBACK TO INFLAMMATION, ASCRIBING INFLAMMATORY REMODELING TO ONE OF THE DRIVERS IN CHRONIC PATHOLOGIES. IN THIS REVIEW, WE DISCUSS THE SUBSTRATE-SPECIFIC READ THROUGH OGG1 HAS EVOLVED IN REGULATING THE INNATE IMMUNE RESPONSE, CONTROLLING ADAPTATIONS OF THE AIRWAY TO ENVIRONMENTAL AND INFLAMMATORY INJURY, WITH A FOCUS ON THE READER FUNCTION OF OGG1 IN INITIATION AND PROGRESSION OF EPITHELIAL TO MESENCHYMAL TRANSITIONS IN CHRONIC PULMONARY DISEASE. 2023 17 3202 25 HDAC2 REGULATES ATYPICAL ANTIPSYCHOTIC RESPONSES THROUGH THE MODULATION OF MGLU2 PROMOTER ACTIVITY. HISTONE DEACETYLASES (HDACS) COMPACT CHROMATIN STRUCTURE AND REPRESS GENE TRANSCRIPTION. IN SCHIZOPHRENIA, CLINICAL STUDIES DEMONSTRATE THAT HDAC INHIBITORS ARE EFFICACIOUS WHEN GIVEN IN COMBINATION WITH ATYPICAL ANTIPSYCHOTICS. HOWEVER, THE MOLECULAR MECHANISM THAT INTEGRATES A BETTER RESPONSE TO ANTIPSYCHOTICS WITH CHANGES IN CHROMATIN STRUCTURE REMAINS UNKNOWN. HERE WE FOUND THAT CHRONIC ATYPICAL ANTIPSYCHOTICS DOWNREGULATED THE TRANSCRIPTION OF METABOTROPIC GLUTAMATE 2 RECEPTOR (MGLU2, ALSO KNOWN AS GRM2), AN EFFECT THAT WAS ASSOCIATED WITH DECREASED HISTONE ACETYLATION AT ITS PROMOTER IN MOUSE AND HUMAN FRONTAL CORTEX. THIS EPIGENETIC CHANGE OCCURRED IN CONCERT WITH A SEROTONIN 5-HT(2A) RECEPTOR-DEPENDENT UPREGULATION AND INCREASED BINDING OF HDAC2 TO THE MGLU2 PROMOTER. VIRALLY MEDIATED OVEREXPRESSION OF HDAC2 IN FRONTAL CORTEX DECREASED MGLU2 TRANSCRIPTION AND ITS ELECTROPHYSIOLOGICAL PROPERTIES, THEREBY INCREASING PSYCHOSIS-LIKE BEHAVIOR. CONVERSELY, HDAC INHIBITORS PREVENTED THE REPRESSIVE HISTONE MODIFICATIONS INDUCED AT THE MGLU2 PROMOTER BY ATYPICAL ANTIPSYCHOTICS, AND AUGMENTED THEIR THERAPEUTIC-LIKE EFFECTS. THESE OBSERVATIONS SUPPORT THE VIEW OF HDAC2 AS A PROMISING NEW TARGET FOR SCHIZOPHRENIA TREATMENT. 2012 18 3864 32 JAK-STAT PATHWAY INHIBITION PARTIALLY RESTORES INTESTINAL HOMEOSTASIS IN HDAC1- AND HDAC2-INTESTINAL EPITHELIAL CELL-DEFICIENT MICE. WE HAVE PREVIOUSLY REPORTED THAT HISTONE DEACETYLASE EPIGENETIC REGULATOR HDAC1 AND HDAC2 DELETION IN INTESTINAL EPITHELIAL CELLS (IEC) DISRUPTS MUCOSAL TISSUE ARCHITECTURE AND BARRIER, CAUSING CHRONIC INFLAMMATION. IN THIS STUDY, PROTEOME AND TRANSCRIPTOME ANALYSIS REVEALED THE IMPORTANCE OF SIGNALING PATHWAYS INDUCED UPON GENETIC IEC-HDAC1 AND HDAC2 DELETION. INDEED, GENE ONTOLOGY BIOLOGICAL PROCESS ANALYSIS OF ENRICHED DEFICIENT IEC RNA AND PROTEINS IDENTIFIED COMMON PATHWAYS, INCLUDING LIPID METABOLIC AND OXIDATION-REDUCTION PROCESS, CELL ADHESION, AND ANTIGEN PROCESSING AND PRESENTATION, RELATED TO IMMUNE RESPONSES, CORRELATING WITH DYSREGULATION OF MAJOR HISTOCOMPATIBILITY COMPLEX (MHC) CLASS II GENES. TOP UPSTREAM REGULATORS INCLUDED REGULATORS ASSOCIATED WITH ENVIRONMENTAL SENSING PATHWAYS TO XENOBIOTICS, MICROBIAL AND DIET-DERIVED LIGANDS, AND ENDOGENOUS METABOLITES. PROTEOME ANALYSIS REVEALED MTOR SIGNALING IEC-SPECIFIC DEFECTS. IN ADDITION TO MTOR, THE STAT AND NOTCH PATHWAYS WERE DYSREGULATED SPECIFICALLY IN JEJUNAL IEC. TO DETERMINE THE IMPACT OF PATHWAY DYSREGULATION ON MUTANT JEJUNUM ALTERATIONS, WE TREATED MUTANT MICE WITH TOFACITINIB, A JAK INHIBITOR. TREATMENT WITH THE INHIBITOR PARTIALLY CORRECTED PROLIFERATION AND TIGHT JUNCTION DEFECTS, AS WELL AS NICHE STABILIZATION BY INCREASING PANETH CELL NUMBERS. THUS, IEC-SPECIFIC HISTONE DEACETYLASES 1 (HDAC1) AND 2 (HDAC2) SUPPORT INTESTINAL HOMEOSTASIS BY REGULATING SURVIVAL AND TRANSLATION PROCESSES, AS WELL AS DIFFERENTIATION AND METABOLIC PATHWAYS. HDAC1 AND HDAC2 MAY PLAY AN IMPORTANT ROLE IN THE REGULATION OF IEC-SPECIFIC INFLAMMATORY RESPONSES BY CONTROLLING, DIRECTLY OR INDIRECTLY, THE JAK/STAT PATHWAY. IEC-SPECIFIC JAK/STAT PATHWAY DEREGULATION MAY BE, AT LEAST IN PART, RESPONSIBLE FOR INTESTINAL HOMEOSTASIS DISRUPTION IN MUTANT MICE. 2021 19 3722 35 INHIBITION OF DNA METHYLATION DURING CHRONIC OBSTRUCTIVE BLADDER DISEASE (COBD) IMPROVES FUNCTION, PATHOLOGY AND EXPRESSION. PARTIAL BLADDER OUTLET OBSTRUCTION DUE TO PROSTATE HYPERPLASIA OR POSTERIOR URETHRAL VALVES, IS A WIDESPREAD CAUSE OF URINARY DYSFUNCTION, PATIENT DISCOMFORT AND ALSO RESPONSIBLE FOR IMMENSE HEALTH CARE COSTS. EVEN AFTER REMOVAL OR RELIEF OF OBSTRUCTION, THE FUNCTIONAL AND PATHOLOGIC ASPECTS OF OBSTRUCTION REMAIN AS A CHRONIC OBSTRUCTIVE BLADDER DISEASE (COBD). EPIGENETIC CHANGES, SUCH AS DNA METHYLATION, CONTRIBUTE TO THE PERSISTENT CHARACTER OF MANY CHRONIC DISEASES, AND MAY BE ALTERED IN COBD. WE TESTED WHETHER CANDIDATE GENES AND PATHWAYS AND THE PATHOPHYSIOLOGY OF COBD WERE AFFECTED BY A HYPOMETHYLATING AGENT, DECITABINE (DAC). COBD WAS CREATED IN FEMALE SPRAGUE-DAWLEY RATS BY SURGICAL LIGATION OF THE URETHRA FOR 6 WEEKS, FOLLOWED BY REMOVAL OF THE SUTURE. SHAM LIGATIONS WERE PERFORMED BY PASSING THE SUTURE BEHIND THE URETHRA. AFTER REMOVAL OF THE OBSTRUCTION OR SHAM REMOVAL, ANIMALS WERE RANDOMIZED TO DAC TREATMENT (1 MG/KG/3-TIMES/WEEK INTRAPERITONEALLY) OR VEHICLE (NORMAL SALINE). BLADDER FUNCTION WAS NON-INVASIVELY TESTED USING METABOLIC CAGES, BOTH ONE DAY PRIOR TO DE-OBSTRUCTION AT 6 WEEKS AND PRIOR TO SACRIFICE AT 10 WEEKS. RESIDUAL VOLUME AND BLADDER MASS WERE MEASURED FOR EACH BLADDER. BLADDERS WERE EXAMINED BY IMMUNOSTAINING AS WELL AS QPCR. THE EFFECTS OF DNA METHYLTRANSFERASE (DNMT)-3A KNOCKOUT OR OVEREXPRESSION ON SMOOTH MUSCLE CELL (SMC) FUNCTION AND PHENOTYPE WERE ALSO EXAMINED IN BLADDER SMC AND EX VIVO CULTURE. RESIDUAL VOLUMES OF THE DAC TREATED GROUP WERE NOT SIGNIFICANTLY DIFFERENT FROM THE NS GROUP. COMPARED TO COBD NS, COBD DAC TREATMENT HELPED PRESERVE MICTURITION VOLUME WITH A SIGNIFICANT RECOVERY OF THE VOIDING EFFICIENCY (RATIO OF THE MAXIMUM VOIDED VOLUME/MAXIMUM BLADDER CAPACITY) BY ONE THIRD (FIG. 1, P > 0.05). BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) VARIANTS 1 AND 5 WERE UPREGULATED BY COBD AND SIGNIFICANTLY REDUCED BY DAC TREATMENT. DEPOSITION OF COLLAGEN IN THE COBD BLADDER WAS REDUCED BY DAC, BUT GROSS HYPERTROPHY REMAINED. IN BLADDER SMC, DNMT3A OVEREXPRESSION LED TO A LOSS OF CONTRACTILE FUNCTION AND PHENOTYPE. IN BLADDERS, PERSISTENTLY ALTERED BY COBD, INHIBITION OF DNA-METHYLATION ENHANCES FUNCTIONAL RECOVERY, UNLIKE TREATMENT DURING PARTIAL OBSTRUCTION, WHICH EXACERBATES OBSTRUCTIVE PATHOLOGY. THE UNDERLYING MECHANISMS MAY RELATE TO THE GENE EXPRESSION CHANGES IN BDNF AND THEIR EFFECTS ON SIGNALING IN THE BLADDER. 2021 20 329 31 ALPHA-OXOGLUTARATE INHIBITS THE PROLIFERATION OF IMMORTALIZED NORMAL BLADDER EPITHELIAL CELLS VIA AN EPIGENETIC SWITCH INVOLVING ARID1A. INTERSTITIAL CYSTITIS (IC) IS A CHRONIC URINARY TRACT DISEASE THAT IS CHARACTERIZED BY UNPLEASANT SENSATIONS, SUCH AS PERSISTENT PELVIC PAIN, IN THE ABSENCE OF INFECTION OR OTHER IDENTIFIABLE CAUSES. WE PREVIOUSLY PERFORMED COMPREHENSIVE METABOLOMICS PROFILING OF URINE SAMPLES FROM IC PATIENTS USING NUCLEAR MAGNETIC RESONANCE AND GAS-CHROMATOGRAPHY/MASS SPECTROMETRY AND FOUND THAT URINARY ALPHA-OXOGLUTARATE (ALPHA-OG), WAS SIGNIFICANTLY ELEVATED. ALPHA-OG, A TRICARBOXYLIC ACID (TCA) CYCLE INTERMEDIATE, REPORTEDLY FUNCTIONS TO SUPPRESS THE PROLIFERATION OF IMMORTALIZED NORMAL HUMAN BLADDER EPITHELIAL CELLS. HERE, WE IDENTIFIED AT-RICH INTERACTIVE DOMAIN 1 A (ARID1A), A KEY CHROMATIN REMODELER, AS BEING HYPOMETHYLATED AND UPREGULATED BY ALPHA-OG TREATMENT. THIS WAS DONE THROUGH EPIC DNA METHYLATION PROFILING AND SUBSEQUENT BIOCHEMICAL APPROACHES, INCLUDING QUANTITATIVE RT-PCR AND WESTERN BLOT ANALYSES. FURTHERMORE, WE FOUND THAT ALPHA-OG ALMOST COMPLETELY SUPPRESSES TEN-ELEVEN TRANSLOCATION (TET) ACTIVITY, BUT DOES NOT AFFECT DNA METHYLTRANSFERASE (DNMT) ACTIVITY. ALTOGETHER, OUR STUDIES REVEAL THE POTENTIAL ROLE OF ALPHA-OG IN EPIGENETIC REMODELING THROUGH ITS EFFECTS ON ARID1A AND TET EXPRESSION IN THE BLADDER. THIS MAY PROVIDE A NEW POSSIBLE THERAPEUTIC STRATEGY IN TREATING IC. 2018