1 4174 138 MELATONIN MEDIATED INHIBITION OF EZH2-NOS2 CROSSTALK ATTENUATES INFLAMMATORY BOWEL DISEASE IN PRECLINICAL IN VITRO AND IN VIVO MODELS. AIMS: INFLAMMATORY BOWEL DISEASE IS CHARACTERISED BY ABDOMINAL PAIN, DIARRHOEA, RECTAL BLEEDING AND WEIGHT LOSS. SOMETIMES IT MAY LEADS TO SEVERE HEALTH COMPLICATIONS RESULTING IN DEATH OF AN INDIVIDUAL. CURRENT RESEARCH EFFORTS TO HIGHLIGHT THE ROLE OF MELATONIN IN REGULATING EZH2, A MASTER EPIGENETIC REGULATOR AND ITS BENEFICIARY EFFECT IN CASE OF IBD MANAGEMENT. MATERIAL METHODS: MURINE MACROPHAGES (RAW 264.7) WERE TREATED WITH LIPOPOLYSACCHARIDES (LPS) TO ACTIVATE THEM FOR GENERATING INFLAMMATORY RESPONSE TO INVESTIGATE EFFICACY OF MELATONIN IN-VITRO MODELS. SIMILARLY, FOR DEVELOPING IN VIVO MODELS, DEXTRAN SODIUM SULPHATE (36-50 KDA) WAS USED. EVALUATIONS OF ANTI-INFLAMMATORY ACTIVITIES WERE CARRIED OUT BY NITRITE ASSAY, WESTERN BLOTTING, Q-PCR, IMMUNOFLUORESCENCE, AND HISTOLOGICAL STUDIES. KEY FINDINGS: REDUCTION OF EPIGENETIC TARGET, EZH2 BY MELATONIN SIGNIFICANTLY IMPROVES THE CLINICAL SYMPTOMS OF DEXTRAN SODIUM SULPHATE INDUCED COLITIS AND MAY BE IMPLICATED AS A POTENTIAL THERAPEUTIC TARGET IN IBD MANAGEMENT. THE PRESENT STUDY EVALUATES THE EFFICACY OF MELATONIN BY EPIGENETIC REGULATION IN IBD MODELS. DOWN REGULATION OF EZH2 BY MELATONIN REDUCED THE CHEMICAL INDUCED INFLAMMATORY INSULTS IN IN VITRO AND IN VIVO MODELS. EXPLORATION OF MOLECULAR PATHWAYS HAS REVEALED INTERLINK OF EZH2 AND NOS2, A HALLMARK OF INFLAMMATION. MOLECULAR MECHANISTIC ACTION OF MELATONIN IS ATTRIBUTED TO INHIBITION OF THE EXPRESSION AND PHYSICAL INTERACTION OF EZH2 AND NOS2. SIGNIFICANCE: OUR STUDY HIGHLIGHTS MELATONIN THERAPEUTIC EFFECT VIA ATTENUATING INTERACTION BETWEEN EZH2 AND NOS2 WHICH IS BENEFICIAL IN MANAGING IBD TREATMENT. 2022 2 386 34 AN IN VIVO IMMUNOHISTOCHEMICAL STUDY ON MACROH2A.1 IN LUNG AND LYMPH-NODE TISSUES EXPOSED TO AN ASBESTIFORM FIBER. AIMS: THE AIM OF THIS STUDY WAS TO INVESTIGATE MACROH2A.1 IMMUNOEXPRESSION IN TISSUES OF SHEEP EXPOSED TO FE. BACKGROUND: THE CORRELATION BETWEEN ASBESTIFORM FIBERS, LUNG CANCER, PLEURAL MESOTHELIOMA, AND OTHER LUNG DISEASES IS ALREADY WELL ESTABLISHED AS THE PATHOPHISIOLOGICAL PATHOPHYSIOLOGICAL RESPIRATORY MECHANISMS INVOLVED BY INHALATION OF FLUORO-EDENITE (FE). THE LATTER IS REPRESENTED BY CELL PROLIFERATION AND INDUCING THE RELEASE OF GROWTH FACTORS, CYTOKINES, AND REACTIVE OXYGEN AND NITRITE SPECIES, WITH DNA DAMAGE THAT CAUSES CHRONIC INFLAMMATION AND CARCINOGENESIS. MACROH2A.1, AND HISTONE VARIANT, SEEMS TO PLAY A ROLE IN SENSING THE METABOLIC STATE OF THE CELL AND LINKING IT WITH CHROMATIN. PHYSIOLOGICALLY, MACROH2A.1 IS EXPRESSED AT LOW LEVELS IN STEM CELLS AND IT BECAME UPREGULATED DURING DIFFERENTIATION, PREVENTING REPROGRAMMING OF INDUCED PLURIPOTENT STEM CELLS AND AFTER NUCLEAR TRANSFER. IN PARTICULAR, MACROH2A.1 HAS BEEN SHOWN TO EXPLICATE A POTENT ANTITUMOR MECHANISM IN VIVO AS IT RESULTS UPREGULATED IN SENESCENT CELLS DETERMINING A PERMANENT GROWTH-ARREST. OBJECTIVE: EVALUATE THE POSSIBLE ROLE OF THE HISTONE VARIANT IN THE ORGANISM IN RESPONSE TO DEEP INSIGHT UNDERSTANDING THE MECHANISMS OF TOXICITY AND THE CELLULAR RESPONSE TO FE. METHODS: LUNG AND LYMPH NODES OF EXPOSED SHEEP WERE SELECTED. SAMPLES WERE PROCESSED FOR HISTOLOGICAL AND IMMUNIHISTOCHEMICAL IMMUNOHISTOCHEMICAL EVALUATIONS. DENSITOMETRIC, MORPHOMETRIC, AND STATISTICAL ANALYSIS ANALYSES WERE CONDUCTED. RESULTS: TISSUE SECTIONS OF FE EXPOSED SHEEP DEMONSTRATED OVEREXPRESSION OF MACROH2A.1 VS UNEXPOSED SAMPLES. THE DATA SUGGEST AN INVOLVEMENT OF THESE THIS MOLECULE IN THE CELLULAR RESPONSE TRIGGERED BY FE DIRECTED EXPOSURE. CONCLUSION: IN THIS CONTEST, MACROH2A.1 OVEREXPRESSION SUPPORTS ITS FUNCTION AS AN EPIGENETIC STABILIZER THAT HELPS TO ESTABLISH AND MAINTAIN DIFFERENTIATED STATES. 2020 3 4044 23 MACROPHAGES IN OXIDATIVE STRESS AND MODELS TO EVALUATE THE ANTIOXIDANT FUNCTION OF DIETARY NATURAL COMPOUNDS. ANTIOXIDANT TESTING OF NATURAL PRODUCTS HAS ATTRACTED INCREASING INTEREST IN RECENT YEARS, MAINLY DUE TO THE FACT THAT AN ANTIOXIDANT-RICH DIET MIGHT PROVIDE HEALTH BENEFITS. ACTIVATED MACROPHAGES ARE A MAJOR SOURCE OF REACTIVE OXYGEN SPECIES, REACTIVE NITROGEN SPECIES, AND PEROXYNITRITE GENERATED THROUGH THE SO-CALLED RESPIRATORY BURST. CONSTITUTIVELY RELEASED PROINFLAMMATORY CYTOKINE, ESPECIALLY TUMOR NECROSIS FACTOR-ALPHA, TRIGGERS NUCLEAR FACTOR-KAPPAB, AND ACTIVATOR PROTEIN-1 TRANSLOCATION LEADING TO THE OVER PRODUCTION OF REACTIVE OXYGEN SPECIES AND REACTIVE NITROGEN SPECIES IN MACROPHAGES. ACTIVATION OF TRANSCRIPTION FACTORS IN THE LONG-LIVED TISSUE-RESIDENT MACROPHAGES AND/OR MONOCYTE-DERIVED MACROPHAGES, TRIGGER EPIGENETIC MODIFICATIONS LEADING TO THE PATHOGENESIS OF CHRONIC DISEASES. NUTRACEUTICALS INCLUDING LIPID RAFT STRUCTURE DISRUPTION AGENT, CHOLESTEROL DEPLETION AGENT, FARNESYLTRANSFERASE INHIBITOR, NUCLEAR FACTOR-KAPPAB BLOCKER (ALPHA,BETA-UNSATURATED CARBONYL COMPOUNDS), GLUCOCORTICOID RECEPTOR AGONIST, AND PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-GAMMA AGONIST HAVE LONG BEEN USED TO INACTIVE MACROPHAGE. THE INHIBITION EFFECTS ON THE FORMATION OF NITRIC OXIDE, SUPEROXIDE, AND NITRITE PEROXIDE MAY BE RESPONSIBLE FOR THE ANTI-INFLAMMATORY FUNCTIONALITIES. ACTIVATED MACROPHAGE MODELS COULD BE USED TO IDENTIFY THE ACTIVE COMPONENTS FOR FUNCTIONAL DIETS DEVELOPMENT THROUGH A MULTIPLE TARGETS STRATEGY. 2017 4 1635 24 DOES A SINGLE BOUT OF EXERCISE IMPACTS BDNF, OXIDATIVE STRESS AND EPIGENETIC MARKERS IN SPINAL CORD INJURY PATIENTS? OUR AIM WAS TO EVALUATE THE IMPACT OF A SINGLE BOUT OF EXERCISE, CONSISTING OF A GAIT TRAINING SESSION WITH BODY WEIGHT SUPPORT (BWS), ON HISTONE ACETYLATION STATUS (GLOBAL HISTONE H4 AND H3 ACETYLATION LEVELS), BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) LEVELS, AND OXIDATIVE STRESS MARKERS IN PERIPHERAL BLOOD OF INDIVIDUALS WITH CHRONIC SPINAL CORD INJURY (SCI). WE ALSO SET OUT TO COMPARE THESE RESPONSES WITH THOSE RECORDED AFTER GAIT TRAINING PERFORMED USING A WALKER AND WITH NO BWS. THE SUBJECTS (NEARLY ALL WITH AN INCOMPLETE SPINAL CORD LESION) WERE EACH SUBMITTED TO TWO 60-MINUTE EXPERIMENTAL SESSIONS ON SEPARATE DAYS WITH A 1- WEEK WASH-OUT PERIOD BETWEEN THE INTERVENTIONS. THE ORDER OF THE SESSIONS WAS RANDOMIZED. BLOOD SAMPLES WERE COLLECTED BEFORE AND AFTER EACH EXPERIMENTAL TRIAL FOR MEASUREMENT OF BIOMARKERS. THE HISTONE ACETYLATION STATUS AND BDNF LEVELS REMAINED UNCHANGED AFTER BOTH INTERVENTIONS. AFTER THE TREADMILL TRAINING, THE PARTICIPANTS SHOWED A STRONG INCREASE IN LEVELS OF OXIDATIVE STRESS MARKERS [PLASMA ADVANCED OXIDATION PROTEIN PRODUCTS (AOPPS), NITRITE AND THIOBARBITURIC ACID-REACTIVE SUBSTANCES] WITHOUT CHANGES IN ANTIOXIDANT MEDIATORS. INSTEAD, ELEVATIONS IN AOPP AND NITRITE CONCENTRATIONS, IN ADDITION TO INCREASED LEVELS OF GLUTATHIONE AND CATALASE ACTIVITY, WERE FOUND AFTER THE WALKER TRAINING. A SINGLE BOUT OF GAIT TRAINING, BE IT CONDUCTED ON A TREADMILL WITH BWS OR USING A WALKER WITHOUT BWS, IS NOT ABLE TO ALTER BDNF LEVELS AND HISTONE ACETYLATION STATUS IN SCI PATIENTS. HOWEVER, THESE TRIALS CAN MODULATE OXIDATIVE STRESS PARAMETERS, SEEMINGLY IN A PROTOCOL-DEPENDENT MANNER. 2019 5 5043 34 PHARMACOKINETICS AND PHARMACODYNAMICS OF CURCUMIN IN REGULATING ANTI-INFLAMMATORY AND EPIGENETIC GENE EXPRESSION. CHRONIC INFLAMMATION IS A KEY DRIVER OF CANCER DEVELOPMENT. NITRITE LEVELS, WHICH ARE REGULATED BY INDUCIBLE NITRIC OXIDE SYNTHASE (INOS), PLAY A CRITICAL ROLE IN INFLAMMATION. WHILE THE ANTI-OXIDANT AND ANTI-INFLAMMATORY EFFECTS OF CURCUMIN, A NATURAL PRODUCT PRESENT IN THE ROOTS OF CURCUMA LONGA HAVE BEEN STUDIED WIDELY, THE ACUTE PHARMACOKINETICS (PK) AND PHARMACODYNAMICS (PD) OF CURCUMIN IN SUPPRESSING PRO-INFLAMMATORY MARKERS AND EPIGENETIC MODULATORS REMAIN UNCLEAR. THIS STUDY EVALUATED THE PK AND PD OF CURCUMIN-INDUCED SUPPRESSION OF LIPOPOLYSACCHARIDE (LPS)-MEDIATED INFLAMMATION IN RAT LYMPHOCYTES. LPS WAS ADMINISTERED INTRAVENOUSLY EITHER ALONE OR WITH CURCUMIN TO FEMALE SPRAGUE-DAWLEY RATS. PLASMA SAMPLES WERE ANALYSED FOR CURCUMIN CONCENTRATION AND MRNA EXPRESSION WAS QUANTIFIED IN LYMPHOCYTES. THE RELATIVE GENE EXPRESSION OF SEVERAL INFLAMMATORY AND EPIGENETIC MODULATORS WAS ANALYSED. TO INVESTIGATE THE RELATIONSHIP BETWEEN CURCUMIN CONCENTRATION AND INOS, TNF-ALPHA, AND IL-6 GENE EXPRESSION, PK/PD MODELING USING JUSKO'S INDIRECT RESPONSE MODEL (IDR) INTEGRATING TRANSIT COMPARTMENTS (TC) DESCRIBING THE DELAYED RESPONSE WAS CONDUCTED. THE CONCENTRATION-TIME PROFILE OF CURCUMIN EXHIBITED A BI-EXPONENTIAL DECLINE, WHICH WAS WELL DESCRIBED BY A TWO-COMPARTMENTAL PHARMACOKINETIC MODEL. IMPORTANTLY THE RESULTS DEMONSTRATE THAT LPS INDUCED GENE EXPRESSION OF PRO-INFLAMMATORY MARKERS IN LYMPHOCYTES, WITH PEAK EXPRESSION AT APPROXIMATELY 3 H AND CURCUMIN SUPPRESSED THE GENE EXPRESSION IN ANIMALS ADMINISTERED WITH LPS. THESE EFFECTS WERE WELL CAPTURED USING THE IDR MODEL AND AN IDR MODEL WITH THE TRANSIT COMPARTMENTS. IN SUMMARY, THE PK/PD MODELING APPROACH COULD POTENTIALLY PROVIDE A ROBUST QUANTITATIVE FRAMEWORK FOR EVALUATING THE ACUTE ANTI-INFLAMMATORY AND EPIGENETIC EFFECTS OF CURCUMIN IN FUTURE CLINICAL TRIALS. 2018 6 6456 30 THYMOSIN BETA4 PREVENTS OXIDATIVE STRESS, INFLAMMATION, AND FIBROSIS IN ETHANOL- AND LPS-INDUCED LIVER INJURY IN MICE. THYMOSIN BETA 4 (TBETA4), AN ACTIN-SEQUESTERING PROTEIN, IS INVOLVED IN TISSUE DEVELOPMENT AND REGENERATION. IT PREVENTS INFLAMMATION AND FIBROSIS IN SEVERAL TISSUES. WE INVESTIGATED THE ROLE OF TBETA4 IN CHRONIC ETHANOL- AND ACUTE LIPOPOLYSACCHARIDE- (LPS-) INDUCED MOUSE LIVER INJURY. C57BL/6 MICE WERE FED 5% ETHANOL IN LIQUID DIET FOR 4 WEEKS PLUS BINGE ETHANOL (5 G/KG, GAVAGE) WITH OR WITHOUT LPS (2 MG/KG, INTRAPERITONEAL) FOR 6 HOURS. TBETA4 (1 MG/KG, INTRAPERITONEAL) WAS ADMINISTERED FOR 1 WEEK. WE DEMONSTRATED THAT TBETA4 PREVENTED ETHANOL- AND LPS-MEDIATED INCREASE IN LIVER INJURY MARKERS AS WELL AS CHANGES IN LIVER PATHOLOGY. IT ALSO PREVENTED ETHANOL- AND LPS-MEDIATED INCREASE IN OXIDATIVE STRESS BY DECREASING ROS AND LIPID PEROXIDATION AND INCREASING THE ANTIOXIDANTS, REDUCED GLUTATHIONE AND MANGANESE-DEPENDENT SUPEROXIDE DISMUTASE. IT ALSO PREVENTED THE ACTIVATION OF NUCLEAR FACTOR KAPPA B BY BLOCKING THE PHOSPHORYLATION OF THE INHIBITORY PROTEIN, IKAPPAB, THEREBY PREVENTED PROINFLAMMATORY CYTOKINE PRODUCTION. MOREOVER, TBETA4 PREVENTED FIBROGENESIS BY SUPPRESSING THE EPIGENETIC REPRESSOR, METHYL-CPG-BINDING PROTEIN 2, THAT COORDINATELY REVERSED THE EXPRESSION OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-GAMMA AND DOWNREGULATED FIBROGENIC GENES, PLATELET-DERIVED GROWTH FACTOR-BETA RECEPTOR, ALPHA-SMOOTH MUSCLE ACTIN, COLLAGEN 1, AND FIBRONECTIN, RESULTING IN REDUCED FIBROSIS. OUR DATA SUGGEST THAT TBETA4 HAS ANTIOXIDANT, ANTI-INFLAMMATORY, AND ANTIFIBROTIC POTENTIAL DURING ALCOHOLIC LIVER INJURY. 2018 7 5760 33 SOLUBLE URIC ACID PRIMES TLR-INDUCED PROINFLAMMATORY CYTOKINE PRODUCTION BY HUMAN PRIMARY CELLS VIA INHIBITION OF IL-1RA. OBJECTIVES: THE STUDY OF THE PROINFLAMMATORY ROLE OF URIC ACID HAS FOCUSED ON THE EFFECTS OF ITS CRYSTALS OF MONOSODIUM URATE (MSU). HOWEVER, LITTLE IS KNOWN WHETHER URIC ACID ITSELF CAN DIRECTLY HAVE PROINFLAMMATORY EFFECTS. IN THIS STUDY, WE INVESTIGATE THE PRIMING EFFECTS OF URIC ACID EXPOSURE ON THE CYTOKINE PRODUCTION OF PRIMARY HUMAN CELLS UPON STIMULATION WITH GOUT-RELATED STIMULI. METHODS: PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) WERE HARVESTED FROM PATIENTS WITH GOUT AND HEALTHY VOLUNTEERS. CELLS WERE PRETREATED WITH OR WITHOUT URIC ACID IN SOLUBLE FORM FOR 24 H AND THEN STIMULATED FOR 24 H WITH TOLL-LIKE RECEPTOR (TLR)2 OR TLR4 LIGANDS IN THE PRESENCE OR ABSENCE OF MSU CRYSTALS. CYTOKINE PRODUCTION WAS MEASURED BY ELISA; MRNA LEVELS WERE ASSESSED USING QPCR. RESULTS: THE PRODUCTION OF INTERLEUKIN (IL)-1BETA AND IL-6 WAS HIGHER IN PATIENTS COMPARED WITH CONTROLS AND THIS CORRELATED WITH SERUM URATE LEVELS. PROINFLAMMATORY CYTOKINE PRODUCTION WAS SIGNIFICANTLY POTENTIATED WHEN CELLS FROM HEALTHY SUBJECTS WERE PRETREATED WITH URIC ACID. SURPRISINGLY, THIS WAS ASSOCIATED WITH A SIGNIFICANT DOWNREGULATION OF THE ANTI-INFLAMMATORY CYTOKINE IL-1 RECEPTOR ANTAGONIST (IL-1RA). THIS EFFECT WAS SPECIFIC TO STIMULATION BY URIC ACID AND WAS EXERTED AT THE LEVEL OF GENE TRANSCRIPTION. EPIGENETIC REPROGRAMMING AT THE LEVEL OF HISTONE METHYLATION BY URIC ACID WAS INVOLVED IN THIS EFFECT. CONCLUSIONS: IN THIS STUDY WE DEMONSTRATE A MECHANISM THROUGH WHICH HIGH CONCENTRATIONS OF URIC ACID (UP TO 50 MG/DL) INFLUENCE INFLAMMATORY RESPONSES BY FACILITATING IL-1BETA PRODUCTION IN PBMCS. WE SHOW THAT A MECHANISM FOR THE AMPLIFICATION OF IL-1BETA CONSISTS IN THE DOWNREGULATION OF IL-1RA AND THAT THIS EFFECT COULD BE EXERTED VIA EPIGENETIC MECHANISMS SUCH AS HISTONE METHYLATION. HYPERURICAEMIA CAUSES A SHIFT IN THE IL-1BETA/IL-1RA BALANCE PRODUCED BY PBMCS AFTER EXPOSURE TO MSU CRYSTALS AND TLR-MEDIATED STIMULI, AND THIS PHENOMENON IS LIKELY TO REINFORCE THE ENHANCED STATE OF CHRONIC INFLAMMATION. 2016 8 5480 31 RESVERATROL REVERSES MORPHINE-INDUCED NEUROINFLAMMATION IN MORPHINE-TOLERANT RATS BY REVERSAL HDAC1 EXPRESSION. BACKGROUND/PURPOSE: WE PREVIOUSLY SHOWED THAT SUBSEQUENT INTRATHECAL (I.T.) INJECTION OF RESVERATROL (30 MUG) SIGNIFICANTLY REVERSES MORPHINE-EVOKED NEUROINFLAMMATION IN MORPHINE-TOLERANT RATS. THE PRESENT STUDY EXAMINED THE UNDERLYING MECHANISM. METHODS: MALE WISTAR RATS WERE IMPLANTED WITH TWO I.T. CATHETERS, ONE OF WHICH WAS CONNECTED TO A MINIOSMOTIC PUMP AND USED FOR MORPHINE (15 MUG/H) OR SALINE INFUSION FOR 120 HOURS. TO EXAMINE THE EFFECTS ON SPINAL CORD EXPRESSION OF HISTONE DEACETYLASE 1 (HDAC1), THE INFLAMMATORY CYTOKINE TUMOR NECROSIS FACTOR-ALPHA (TNF-ALPHA), AND TNF RECEPTOR (TNFR) 1 AND TNFR2 DURING TOLERANCE INDUCTION, A TAIL-FLICK TEST WAS PERFORMED PRIOR TO INFUSION AND AFTER 24 HOURS, 48 HOURS, 72 HOURS, 96 HOURS, AND 120 HOURS OF INFUSION. RESULTS: RESVERATROL TREATMENT PRIOR TO MORPHINE CHALLENGE RESTORED THE ANTINOCICEPTIVE EFFECT OF MORPHINE IN MORPHINE-TOLERANT RATS AND REVERSED THE MORPHINE INFUSION-INDUCED INCREASE IN HDAC1, TNF-ALPHA, AND TNFR1 EXPRESSION. MOREOVER, CHRONIC MORPHINE INFUSION INCREASED TNFR1-SPECIFIC EXPRESSION IN NEURON IN MORPHINE-TOLERANT RAT SPINAL CORDS, AND THIS EFFECT WAS ALMOST COMPLETELY INHIBITED BY RESVERATROL TREATMENT PRIOR TO MORPHINE CHALLENGE. CONCLUSION: RESVERATROL RESTORES THE ANTINOCICEPTIVE EFFECT OF MORPHINE BY REVERSING MORPHINE INFUSION-INDUCED SPINAL CORD NEUROINFLAMMATION AND INCREASE IN TNFR1 EXPRESSION. THE REVERSAL OF THE MORPHINE-INDUCED INCREASE IN TNFR1 EXPRESSION BY RESVERATROL IS PARTIALLY DUE TO REVERSAL OF THE MORPHINE INFUSION-INDUCED INCREASE IN HDAC1 EXPRESSION. RESVERATROL PRETREATMENT CAN BE USED AS AN ADJUVANT IN CLINICAL PAIN MANAGEMENT FOR PATIENTS WHO NEED LONG-TERM MORPHINE TREATMENT OR WITH NEUROPATHIC PAIN. 2016 9 5868 36 SUPPRESSIVE EFFECTS OF METFORMIN ON T-HELPER 1-RELATED CHEMOKINES EXPRESSION IN THE HUMAN MONOCYTIC LEUKEMIA CELL LINE THP-1. PURPOSE OF THE STUDY: TYPE 1 AND TYPE 2 DIABETES MELLITUS (DM) ARE CHRONIC T-CELL-MEDIATED INFLAMMATORY DISEASES. METFORMIN IS A WIDELY USED DRUG FOR TYPE 2 DM THAT REDUCES THE NEED FOR INSULIN IN TYPE 1 DM. HOWEVER, WHETHER METFORMIN HAS AN ANTI-INFLAMMATORY EFFECT FOR TREATING DM IS UNKNOWN. WE INVESTIGATED THE ANTI-INFLAMMATORY MECHANISM OF METFORMIN IN THE HUMAN MONOCYTIC LEUKEMIA CELL LINE THP-1. MATERIALS AND METHODS: THE HUMAN MONOCYTIC LEUKEMIA CELL LINE THP-1 WAS PRETREATED WITH METFORMIN AND STIMULATED WITH LIPOPOLYSACCHARIDE (LPS). THE PRODUCTION OF T-HELPER (TH)-1-RELATED CHEMOKINES INCLUDING INTERFERON-GAMMA-INDUCED PROTEIN-10 (IP-10) AND MONOCYTE CHEMOATTRACTANT PROTEIN-1 (MCP-1), TH2-RELATED CHEMOKINE MACROPHAGE-DERIVED CHEMOKINE, AND THE PROINFLAMMATORY CHEMOKINE TUMOR NECROSIS FACTOR-ALPHA WAS MEASURED USING ENZYME-LINKED IMMUNOSORBENT ASSAY. INTRACELLULAR SIGNALING PATHWAYS WERE INVESTIGATED USING WESTERN BLOT ANALYSIS AND CHROMATIN IMMUNOPRECIPITATION ASSAY. RESULTS: METFORMIN SUPPRESSED LPS-INDUCED IP-10 AND MCP-1 PRODUCTION AS WELL AS LPS-INDUCED PHOSPHORYLATION OF C-JUN N-TERMINAL KINASE (JNK), P38, EXTRACELLULAR SIGNAL-REGULATED KINASE (ERK), AND NUCLEAR FACTOR-KAPPA B (NF-KAPPAB). MOREOVER, METFORMIN SUPPRESSED LPS-INDUCED ACETYLATION OF HISTONES H3 AND H4 AT THE IP-10 PROMOTER. CONCLUSIONS: METFORMIN SUPPRESSED THE PRODUCTION OF TH1-RELATED CHEMOKINES IP-10 AND MCP-1 IN THP-1 CELLS. SUPPRESSIVE EFFECTS OF METFORMIN ON IP-10 PRODUCTION MIGHT BE ATTRIBUTED AT LEAST PARTIALLY TO THE JNK, P38, ERK, AND NF-KAPPAB PATHWAYS AS WELL AS TO EPIGENETIC REGULATION THROUGH THE ACETYLATION OF HISTONES H3 AND H4. THESE RESULTS INDICATED THE THERAPEUTIC ANTI-INFLAMMATORY POTENTIAL OF METFORMIN. 2018 10 4582 29 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 11 593 38 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 12 6166 29 THE GLUTATHIONE SYSTEM: A NEW DRUG TARGET IN NEUROIMMUNE DISORDERS. GLUTATHIONE (GSH) HAS A CRUCIAL ROLE IN CELLULAR SIGNALING AND ANTIOXIDANT DEFENSES EITHER BY REACTING DIRECTLY WITH REACTIVE OXYGEN OR NITROGEN SPECIES OR BY ACTING AS AN ESSENTIAL COFACTOR FOR GSH S-TRANSFERASES AND GLUTATHIONE PEROXIDASES. GSH ACTING IN CONCERT WITH ITS DEPENDENT ENZYMES, KNOWN AS THE GLUTATHIONE SYSTEM, IS RESPONSIBLE FOR THE DETOXIFICATION OF REACTIVE OXYGEN AND NITROGEN SPECIES (ROS/RNS) AND ELECTROPHILES PRODUCED BY XENOBIOTICS. ADEQUATE LEVELS OF GSH ARE ESSENTIAL FOR THE OPTIMAL FUNCTIONING OF THE IMMUNE SYSTEM IN GENERAL AND T CELL ACTIVATION AND DIFFERENTIATION IN PARTICULAR. GSH IS A UBIQUITOUS REGULATOR OF THE CELL CYCLE PER SE. GSH ALSO HAS CRUCIAL FUNCTIONS IN THE BRAIN AS AN ANTIOXIDANT, NEUROMODULATOR, NEUROTRANSMITTER, AND ENABLER OF NEURON SURVIVAL. DEPLETION OF GSH LEADS TO EXACERBATION OF DAMAGE BY OXIDATIVE AND NITROSATIVE STRESS; HYPERNITROSYLATION; INCREASED LEVELS OF PROINFLAMMATORY MEDIATORS AND INFLAMMATORY POTENTIAL; DYSFUNCTIONS OF INTRACELLULAR SIGNALING NETWORKS, E.G., P53, NUCLEAR FACTOR-KAPPAB, AND JANUS KINASES; DECREASED CELL PROLIFERATION AND DNA SYNTHESIS; INACTIVATION OF COMPLEX I OF THE ELECTRON TRANSPORT CHAIN; ACTIVATION OF CYTOCHROME C AND THE APOPTOTIC MACHINERY; BLOCKADE OF THE METHIONINE CYCLE; AND COMPROMISED EPIGENETIC REGULATION OF GENE EXPRESSION. AS SUCH, GSH DEPLETION HAS MARKED CONSEQUENCES FOR THE HOMEOSTATIC CONTROL OF THE IMMUNE SYSTEM, OXIDATIVE AND NITROSATIVE STRESS (O&NS) PATHWAYS, REGULATION OF ENERGY PRODUCTION, AND MITOCHONDRIAL SURVIVAL AS WELL. GSH DEPLETION AND CONCOMITANT INCREASE IN O&NS AND MITOCHONDRIAL DYSFUNCTIONS PLAY A ROLE IN THE PATHOPHYSIOLOGY OF DIVERSE NEUROIMMUNE DISORDERS, INCLUDING DEPRESSION, MYALGIC ENCEPHALOMYELITIS/CHRONIC FATIGUE SYNDROME AND PARKINSON'S DISEASE, SUGGESTING THAT DEPLETED GSH IS AN INTEGRAL PART OF THESE DISEASES. THERAPEUTICAL INTERVENTIONS THAT AIM TO INCREASE GSH CONCENTRATIONS IN VIVO INCLUDE N-ACETYL CYSTEINE; NRF-2 ACTIVATION VIA HYPERBARIC OXYGEN THERAPY; DIMETHYL FUMARATE; PHYTOCHEMICALS, INCLUDING CURCUMIN, RESVERATROL, AND CINNAMON; AND FOLATE SUPPLEMENTATION. 2014 13 440 25 ANTIOXIDATIVE EFFECTS OF RHODIOLA GENUS: PHYTOCHEMISTRY AND PHARMACOLOGICAL MECHANISMS AGAINST THE DISEASES. RHODIOLA AS ONE OF TRADITIONAL MEDICINES HAS BEEN USED FOR CLINICAL TREATMENTS DUE TO ITS STRONG ANTIOXIDANT PROPERTIES. PHYTOCHEMICAL ANALYSIS REVEALED THE PRESENCE OF FLAVONOIDS, PHENYLPROPANOIDS, PHENYLETHANOL/BENZYL ALCOHOL DERIVATIVES, CYANOGENIC GLYCOSIDES AND TERPENOIDS. THE BIOACTIVE COMPOUNDS HAD BEEN DEMONSTRATED TO BE EFFECTIVE AT SCAVENGING REACTIVE OXYGEN SPECIES (ROS). THE STRUCTURES CONTAIN PHENOLIC HYDROXYL GROUPS AND UNSATURATED BONDS. THIS ARTICLE REVIEWS ANTIOXIDANT CAPACITIES OF THE EXTRACTS AND BIOACTIVE COMPONENTS DERIVED FROM RHODIOLA PLANTS. AS THE MAJOR PHARMACOLOGICAL INGREDIENT, SALIDROSIDE IS RIGOROUSLY INVESTIGATED AND USED IN SCIENTIFIC RESEARCHES AND CLINICAL PRACTICES. ACCUMULATED EVIDENCES INDICATED THAT EXTRACTS OF RHODIOLA PLANTS OR SALIDROSIDE COULD BE ABLE TO REVERSE DNA DAMAGE AND ALTER EXPRESSION OF CYTOKINES AND ANTIOXIDATIVE ENZYMES INDUCED BY ROS. THE UNDERLYING MECHANISMS FOR THE ANTIOXIDATIVE EFFECTS OF THE HERB HAVE BEEN INVESTIGATED IN THE LAST TWO DECADES. WE SUMMARIZE THE POSSIBLE EFFECTS AND ACTING PATHWAYS FOR THE HERB INVOLVED IN SEVERAL CHRONIC DISEASES IN CARDIOVASCULAR, RESPIRATORY, AND NERVOUS SYSTEMS, AS WELL AS POTENTIAL EPIGENETIC INFLUENCES. THE INFORMATION GENERATED FROM EXPERIMENTAL AND CLINICAL STUDIES OFFERED VALUABLE INSIGHTS FOR FURTHER INVESTIGATIONS OF MEDICAL POTENTIALS OF RHODIOLA PLANTS. 2017 14 5448 28 REPRESSION OF HDAC5 BY ACETATE RESTORES HYPOTHALAMIC-PITUITARY-OVARIAN FUNCTION IN TYPE 2 DIABETES MELLITUS. TYPE 2 DIABETES MELLITUS (T2DM) ACCOUNTS FOR 90-95 % OF WORLDWIDE DIABETES CASES AND IS PRIMARILY CHARACTERIZED BY INSULIN RESISTANCE. ITS PROGRESSION AS A CHRONIC METABOLIC DISEASE HAS BEEN LARGELY ASSOCIATED WITH FEMALE REPRODUCTIVE ABNORMALITIES, INCLUDING OVARIAN DYSFUNCTION WITH CONSEQUENT INFERTILITY. EPIGENETIC MODIFICATIONS HAVE BEEN SUGGESTED AS A POSSIBLE LINK TO METABOLIC COMORBIDITIES. WE THEREFORE HYPOTHESIZED THAT SHORT CHAIN FATTY ACIDS, ACETATE (ACA), A POTENTIAL HISTONE DEACETYLASE INHIBITOR (HDAC) AMELIORATES HYPOTHALAMIC-PITUITARY-OVARIAN (HPO) DYSFUNCTION IN T2DM. FEMALE WISTAR RATS WEIGHING 160-190 G WERE ALLOTTED INTO THREE GROUPS (N = 6/GROUP): CONTROL (VEHICLE; PO), T2D AND T2D + ACA (200 MG/KG; PO). T2DM WAS INDUCED BY FRUCTOSE ADMINISTRATION (10 %; W/V) FOR 6 WEEKS AND SINGLE DOSE OF STREPTOZOTOCIN (35 MG/KG; IP). THE PRESENT DATA SHOWED THAT IN ADDITION TO INSULIN RESISTANCE, INCREASED FASTING BLOOD GLUCOSE AND INSULIN, T2DM INDUCED ELEVATED PLASMA, HYPOTHALAMIC AND OVARIAN TRIGLYCERIDE, LIPID PEROXIDATION, TNF-ALPHA AND GLUTATHIONE DEPLETION. ASIDE, T2DM ALSO LED TO INCREASED PLASMA LACTATE PRODUCTION AND GAMMA-GLUTAMYL TRANSFERASE AS WELL AS DECREASED GONADOTROPINS/17BETA-ESTRADIOL. HISTOLOGICALLY, HYPOTHALAMUS, PITUITARY AND OVARIES REVEALED DISRUPTED NEURONAL CELLS/MODERATE HEMORRHAGE, ALTERED MORPHOLOGY/VASCULAR CONGESTIONS, AND DEGENERATED ANTRAL FOLLICLE/GRAAFIAN FOLLICLE WITH MILD FIBROSIS AND INFILTRATED INFLAMMATORY CELLS RESPECTIVELY IN T2D ANIMALS. INTERESTINGLY, THESE ALTERATIONS WERE ACCOMPANIED BY ELEVATED PLASMA/HYPOTHALAMIC HDAC5 AND ATTENUATED WHEN TREATED WITH ACETATE. THE PRESENT RESULTS DEMONSTRATE THAT T2DM INDUCES HPO DYSFUNCTION, WHICH IS ACCOMPANIED BY ELEVATED CIRCULATING/HYPOTHALAMIC HDAC5. THE RESULTS IN ADDITION SUGGEST THAT ACETATE RESTORES HPO FUNCTION IN T2DM BY SUPPRESSION OF HDAC5 AND ENHANCEMENT OF INSULIN SENSITIVITY. 2021 15 2716 22 EXERCISE-MODULATED EPIGENETIC MARKERS AND INFLAMMATORY RESPONSE IN COPD INDIVIDUALS: A PILOT STUDY. THE STUDY INVESTIGATED THE EFFECTS OF EXERCISE ON EPIGENETIC SIGNALS AND SYSTEMIC CYTOKINE LEVELS IN CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) INDIVIDUALS. TEN PARTICIPANTS OF A PULMONARY REHABILITATION PROGRAM WERE SUBMITTED TO 24 SESSIONS OF A SUPERVISIONED EXERCISE PROTOCOL THRICE-WEEKLY (90MIN/SESSION). BLOOD SAMPLES WERE COLLECTED AT BASELINE, AFTER THE 1ST SESSION, BEFORE AND AFTER THE 24TH SESSION. A DNA HYPOMETHYLATION STATUS WAS OBSERVED AFTER THE 1ST SESSION WHEN COMPARED AT BASELINE, WHILE GLOBAL HISTONE H4 ACETYLATION STATUS WAS UNALTERED IN ANY TIME-POINTS EVALUATED. NO SIGNIFICANT CHANGES WERE OBSERVED ON CYTOKINE LEVELS AFTER THE 1ST SESSION. A SIGNIFICANT ENHANCEMENT ON INTERLEUKIN 6 (IL-6) AND A DECREASE ON TRANSFORMING GROWTH FACTOR-BETA (TGF-BETA) LEVELS WERE FOUND AFTER THE 24TH SESSION WHEN COMPARED TO THE PRE 24TH SESSION. MOREOVER, 23 SESSIONS OF EXERCISE WERE ABLE TO DIMINISH SIGNIFICANTLY THE BASAL LEVELS OF IL-6 AND INTERLEUKIN 8 (IL-8). THESE DATA SUGGEST A POTENTIAL ROLE OF EPIGENETIC MACHINERY IN MEDIATING THE ANTI-INFLAMMATORY EFFECTS OF EXERCISE IN COPD PATIENTS. 2017 16 1105 39 COMBINED INHIBITION OF HISTONE DEACETYLASES AND BET FAMILY PROTEINS AS EPIGENETIC THERAPY FOR NERVE INJURY-INDUCED NEUROPATHIC PAIN. CURRENT TREATMENTS FOR NEUROPATHIC PAIN HAVE OFTEN MODERATE EFFICACY AND PRESENT UNWANTED EFFECTS SHOWING THE NEED TO DEVELOP EFFECTIVE THERAPIES. ACCUMULATING EVIDENCE SUGGESTS THAT HISTONE ACETYLATION PLAYS ESSENTIAL ROLES IN CHRONIC PAIN AND THE ANALGESIC ACTIVITY OF HISTONE DEACETYLASES (HDACS) INHIBITORS IS DOCUMENTED. BROMODOMAIN AND EXTRA-TERMINAL DOMAIN (BET) PROTEINS ARE EPIGENETIC READERS THAT INTERACT WITH ACETYLATED LYSINE RESIDUES ON HISTONES, BUT LITTLE IS KNOWN ABOUT THEIR IMPLICATION IN NEUROPATHIC PAIN. THUS, THE CURRENT STUDY WAS AIMED TO INVESTIGATE THE EFFECT OF THE COMBINATION OF HDAC AND BET INHIBITORS IN THE SPARED NERVE INJURY (SNI) MODEL IN MICE. INTRANASAL ADMINISTRATION OF I-BET762 (BET INHIBITOR) OR SAHA (HDAC INHIBITOR) ATTENUATED THERMAL AND MECHANICAL HYPERSENSITIVITY AND THIS ANTIALLODYNIC ACTIVITY WAS IMPROVED BY CO-ADMINISTRATION OF BOTH DRUGS. SPINAL CORD SECTIONS OF SNI MICE SHOWED AN INCREASED EXPRESSION OF HDAC1 AND BRD4 PROTEINS AND COMBINATION PRODUCED A STRONGER REDUCTION COMPARED TO EACH EPIGENETIC AGENT ALONE. SAHA AND I-BET762, ADMINISTERED ALONE OR IN COMBINATION, COUNTERACTED THE SNI-INDUCED MICROGLIA ACTIVATION BY INHIBITING THE EXPRESSION OF IBA1, CD11B, INDUCIBLE NITRIC OXIDE SYNTHASE (INOS), THE ACTIVATION OF NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) AND SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION-1 (STAT1) WITH COMPARABLE EFFICACY. CONVERSELY, THE EPIGENETIC INHIBITORS SHOWED A MODEST EFFECT ON SPINAL PROINFLAMMATORY CYTOKINES CONTENT THAT WAS SIGNIFICANTLY POTENTIATED BY THEIR COMBINATION. PRESENT RESULTS INDICATE A KEY ROLE OF ACETYLATED HISTONES AND THEIR RECRUITMENT BY BET PROTEINS ON MICROGLIA-MEDIATED SPINAL NEUROINFLAMMATION. TARGETING NEUROPATHIC PAIN WITH THE COMBINATION OF HDAC AND BET INHIBITORS MAY REPRESENT A PROMISING NEW THERAPEUTIC OPTION. 2021 17 5055 28 PHASE 2 TRIAL OF THE HISTONE DEACETYLASE INHIBITOR ROMIDEPSIN FOR THE TREATMENT OF REFRACTORY MULTIPLE MYELOMA. BACKGROUND: EPIGENETIC DYSREGULATION IS A HALLMARK OF CANCER, INCLUDING MULTIPLE MYELOMA. INHIBITORS OF HISTONE DEACETYLASES (HDACS) INDUCE DNA HYPERACETYLATION BY INHIBITING REMOVAL OF ACETYL GROUPS FROM AMINO TAILS ON HISTONE PROTEINS, THEREBY UNCOILING CONDENSED CHROMATIN FAVORING TRANSCRIPTION OF SILENCED GENES, INCLUDING TUMOR SUPPRESSOR GENES. ROMIDEPSIN IS AN HDAC INHIBITOR THAT EXHIBITS ANTIPROLIFERATIVE AND APOPTOTIC EFFECTS AGAINST MULTIPLE MYELOMA CELL LINES. METHODS: A PHASE 2 TRIAL WAS PERFORMED OF ROMIDEPSIN IN PATIENTS WITH MULTIPLE MYELOMA WHO WERE REFRACTORY TO STANDARD THERAPY. TREATMENT WAS COMPRISED OF ROMIDEPSIN (13 MG/M(2)) GIVEN AS A 4-HOUR INTRAVENOUS INFUSION ON DAYS 1, 8, AND 15 EVERY 28 DAYS). THIRTEEN PATIENTS RECEIVED A MEDIAN OF 2 CYCLES OF THERAPY (RANGE, 1-7 CYCLES). RESULTS: ALTHOUGH NO PATIENTS HAD AN OBJECTIVE RESPONSE, 4 OF 12 PATIENTS WITH SECRETORY MYELOMA EXHIBITED EVIDENCE OF M-PROTEIN STABILIZATION, AND SEVERAL OTHER PATIENTS EXPERIENCED IMPROVEMENT IN BONE PAIN AND RESOLUTION OF HYPERCALCEMIA. CONCLUSIONS: THE RESULTS OF THE CURRENT STUDY DEMONSTRATE THAT ROMIDEPSIN, AS A SINGLE AGENT, IS UNLIKELY TO BE ASSOCIATED WITH A RESPONSE RATE OF >/=30% IN PATIENTS WITH REFRACTORY MYELOMA, ALTHOUGH THERE WAS SOME CLINICAL EVIDENCE SUGGESTING A BIOLOGICAL EFFECT ASSOCIATED WITH THERAPY. 2011 18 5010 28 PEROXIDATION OF LINOLEIC, ARACHIDONIC AND OLEIC ACID IN RELATION TO THE INDUCTION OF OXIDATIVE DNA DAMAGE AND CYTOGENETIC EFFECTS. IN THE PRESENT STUDY, THE POSSIBLE ROLE OF THE POLYUNSATURATED FATTY ACIDS LINOLEIC AND ARACHIDONIC ACID IN THE CHEMICAL INDUCTION OF CARCINOGENESIS HAS BEEN INVESTIGATED. ANALYSIS OF 7,8-DIHYDRO-8-OXO-2'-DEOXYGUANOSINE (8-OXODG) LEVELS IN 2'-DEOXYGUANOSINE (DG) AND ISOLATED DNA HAS DEMONSTRATED THAT LINOLEIC AND ARACHIDONIC ACID ARE CAPABLE OF INDUCING THIS SPECIFIC GENOTOXIC DAMAGE. THIS EFFECT APPEARS TO BE RELATED TO THE DEGREE OF FATTY ACID UNSATURATION, SINCE IT WAS NOT INDUCED BY MONOUNSATURATED OLEIC ACID. ENZYMATIC PEROXIDATION OF LINOLEIC AND ARACHIDONIC ACID RESULTED IN A SIGNIFICANT INCREASE IN OXIDATIVE DNA DAMAGE. STUDIES ON THE INTERFERENCE OF RADICAL SCAVENGERS WITH THE INDUCTION OF 8-OXODG IN COMBINATION WITH ELECTRON SPIN RESONANCE SPECTROSCOPY DEMONSTRATED THAT THE SUPEROXIDE ANION WAS GENERATED DURING PEROXIDATION OF THESE FATTY ACIDS AND THAT SINGLET OXYGEN IS MOST LIKELY INVOLVED IN THE FORMATION OF OXIDATIVE DNA DAMAGE. THE LEVEL OF OXIDATIVE DAMAGE IN DG AND SINGLE-STRANDED DNA WAS HIGHER AS COMPARED TO THAT IN NATIVE DNA AFTER EQUIMOLAR TREATMENT. EXPOSURE OF HUMAN LYMPHOCYTES TO LINOLEIC OR ARACHIDONIC ACID DID NOT RESULT IN A SIGNIFICANT INCREASE IN LEVELS OF 8-OXODG. THIS MAY INDICATE THAT THE RATE OF INTRACELLULAR PEROXIDATION IS RELATIVELY LOW AND/OR THAT NUCLEAR DNA IN INTACT CELLS IS EFFECTIVELY PROTECTED AGAINST GENETIC DAMAGE INDUCED BY REACTIVE OXYGEN SPECIES. IT IS THEREFORE CONCLUDED THAT RELATIVELY SHORT PERIODS OF LINOLEIC OR ARACHIDONIC ACID ADMINISTRATION ARE NOT LIKELY TO IMPOSE A DIRECT GENOTOXIC RISK. IT CAN, HOWEVER, NOT BE EXCLUDED THAT CHRONIC EXPOSURE TO POLYUNSATURATED FATTY ACIDS INDUCES OXIDATIVE DNA DAMAGE OR IS RELATED TO CANCER RISK BY EPIGENETIC MECHANISMS, AS IS ALSO INDICATED BY THE OBSERVED CYTOTOXIC EFFECTS OF LINOLEIC AND ARACHIDONIC ACID. 1994 19 5074 34 PHYSIOLOGIC AND EPIGENETIC EFFECTS OF NUTRIENTS ON DISEASE PATHWAYS. BACKGROUND/OBJECTIVES: EPIGENETIC REGULATION BY NUTRIENTS CAN INFLUENCE THE DEVELOPMENT OF SPECIFIC DISEASES. THIS STUDY SOUGHT TO EXAMINE THE EFFECT OF INDIVIDUAL NUTRIENTS AND NUTRIENT FAMILIES IN THE CONTEXT OF PREVENTING CHRONIC METABOLIC DISEASES VIA EPIGENETIC REGULATION. THE INHIBITION OF LIPID ACCUMULATION AND INFLAMMATION BY NUTRIENTS INCLUDING PROTEINS, LIPIDS, VITAMINS, AND MINERALS WERE OBSERVED, AND HISTONE ACETYLATION BY HISTONE ACETYLTRANSFERASE (HAT) WAS MEASURED. CORRELATIVE ANALYSES WERE ALSO PERFORMED. MATERIALS/METHODS: NUTRIENTS WERE SELECTED ACCORDING TO INFORMATION FROM THE KOREAN MINISTRY OF FOOD AND DRUG SAFETY. SELECTED NUTRIENT FUNCTIONALITIES, INCLUDING THE ATTENUATION OF FATTY ACID-INDUCED LIPID ACCUMULATION AND LIPOPOLYSACCHARIDE-MEDIATED ACUTE INFLAMMATION WERE EVALUATED IN MOUSE MACROPHAGE RAW264.7 AND MOUSE HEPATOCYTE AML-12 CELLS. EFFECTS OF THE SELECTED NUTRIENTS ON IN VITRO HAT INHIBITION WERE ALSO EVALUATED. RESULTS: NITRIC OXIDE (NO) PRODUCTION CORRELATED WITH HAT ACTIVITY, WHICH WAS REGULATED BY THE AMINO ACIDS GROUP, SUGGESTING THAT AMINO ACIDS POTENTIALLY CONTRIBUTE TO THE ATTENUATION OF NO PRODUCTION VIA THE INHIBITION OF HAT ACTIVITY. UNSATURATED FATTY ACIDS TENDED TO ATTENUATE INFLAMMATION BY INHIBITING NO PRODUCTION, WHICH MAY BE ATTRIBUTABLE TO THE INHIBITION OF IN VITRO HAT ACTIVITY. IN CONTRAST TO WATER-SOLUBLE VITAMINS, THE LIPID-SOLUBLE VITAMINS SIGNIFICANTLY DECREASED NO PRODUCTION. WATER- AND LIPID-SOLUBLE VITAMINS BOTH EXHIBITED SIGNIFICANT INHIBITORY ACTIVITIES AGAINST HAT. IN ADDITION, CALCIUM AND MANGANESE SIGNIFICANTLY INHIBITED LIPID ACCUMULATION, NO PRODUCTION, AND HAT ACTIVITY. CONCLUSIONS: SEVERAL CANDIDATE NUTRIENTS AND THEIR FAMILY MEMBERS MAY HAVE ROLES IN THE PREVENTION OF DISEASES, INCLUDING HEPATIC STEATOSIS AND INFLAMMATION-RELATED DISEASES (I.E., NONALCOHOLIC STEATOHEPATITIS) VIA EPIGENETIC REGULATION. FURTHER STUDIES ARE WARRANTED TO DETERMINE WHICH SPECIFIC AMINO ACIDS, UNSATURATED FATTY ACIDS AND LIPID-SOLUBLE VITAMINS OR SPECIFIC MINERALS INFLUENCE THE DEVELOPMENT OF STEATOSIS AND INFLAMMATORY-RELATED DISEASES. 2023 20 2822 33 FIRST-IN-HUMAN STUDY OF INHALED AZACITIDINE IN PATIENTS WITH ADVANCED NON-SMALL CELL LUNG CANCER. BACKGROUND: AEROSOLIZED AZACITIDINE HAS BEEN SHOWN TO INHIBIT ORTHOTOPIC LUNG CANCER GROWTH AND INDUCE RE-EXPRESSION OF METHYLATED TUMOR SUPPRESSOR GENES IN MURINE MODELS. WE HYPOTHESIZED THAT INHALED AZACITIDINE IS SAFE AND EFFECTIVE IN REVERSING EPIGENETIC CHANGES IN THE BRONCHIAL EPITHELIUM SECONDARY TO CHRONIC SMOKING. PATIENTS AND METHODS: WE REPORT THE FIRST IN HUMAN STUDY OF INHALED AZACITIDINE. AZACITIDINE IN AQUEOUS SOLUTION WAS USED TO GENERATE AN AEROSOL SUSPENSION OF 0.25-5 MUM PARTICLE SIZE. MAIN INCLUSION CRITERIA: STAGE IV OR RECURRENT NSCLC WITH PREDOMINANTLY LUNG INVOLVEMENT, >/=1 PRIOR SYSTEMIC THERAPY, ECOG PS 0-1, AND ADEQUATE PULMONARY FUNCTION. PATIENTS RECEIVED INHALED AZACITIDINE DAILY ON DAYS 1-5 AND 15-19 OF 28-DAY CYCLES, AT 3 ESCALATING DOSES (15, 30 AND 45 MG/M(2) DAILY). THE PRIMARY OBJECTIVE WAS TO DETERMINE THE FEASIBILITY AND TOLERABILITY OF THIS NEW THERAPEUTIC MODALITY. THE KEY SECONDARY OBJECTIVES INCLUDED PHARMACOKINETICS, METHYLATION PROFILES AND EFFICACY. RESULTS: FROM 3/2015 TO 2/2018, EIGHT PATIENTS RECEIVED A MEDIAN NUMBER OF 2 (IQR = 1) CYCLES OF INHALED AZACITIDINE. NO CLINICALLY SIGNIFICANT ADVERSE EVENTS WERE OBSERVED, EXCEPT ONE PATIENT TREATED AT THE HIGHEST DOSE DEVELOPED AN ASYMPTOMATIC GRADE 2 DECREASED DLCO WHICH RESOLVED SPONTANEOUSLY. ONE PATIENT RECEIVING 12 CYCLES OF THERAPY HAD AN OBJECTIVE AND DURABLE PARTIAL RESPONSE, AND TWO PATIENTS HAD STABLE DISEASE. PLASMA AZACITIDINE WAS ONLY BRIEFLY DETECTABLE IN PATIENTS TREATED AT THE HIGHER DOSES. MOREOVER, IN 2 OF 3 PARTICIPANTS WHO AGREED AND UNDERWENT PRE- AND POST-TREATMENT BRONCHOSCOPY, THE GLOBAL DNA METHYLATION IN THE BRONCHIAL EPITHELIUM DECREASED BY 24 % AND 79 % POST-THERAPY, RESPECTIVELY. THE INTERVAL BETWEEN LAST INHALED TREATMENT AND BRONCHOSCOPY WAS 3 DAYS. CONCLUSIONS: INHALED AZACITIDINE RESULTED IN NEGLIGIBLE PLASMA LEVELS COMPARED TO THE PREVIOUSLY REPORTED SUBCUTANEOUS ADMINISTRATION AND WAS WELL-TOLERATED. THE RESULTS JUSTIFY THE CONTINUED DEVELOPMENT OF INHALED AZACITIDINE AT NON-CYTOTOXIC DOSES FOR PATIENTS WITH LUNG-CONFINED MALIGNANT AND/OR PREMALIGNANT LESIONS. 2021