1 3744 123 INSIGHTS INTO THE CRYSTAL STRUCTURE OF BRD2-BD2 - PHENANTHRIDINONE COMPLEX AND THEORETICAL STUDIES ON PHENANTHRIDINONE ANALOGS. BROMODOMAIN AND EXTRA-TERMINAL FAMILY PROTEINS RECOGNIZE THE ACETYLATED HISTONE CODE ON CHROMATIN AND PARTICIPATE IN DOWNSTREAM PROCESSES LIKE DNA REPLICATION, MODIFICATION, AND REPAIR. AS PART OF EPIGENETIC APPROACHES, BRD2 AND BRD4 WERE IDENTIFIED AS PUTATIVE TARGETS, FOR THE MANAGEMENT OF CHRONIC DISEASES. WE HAVE RECENTLY REPORTED THE DISCOVERY OF A NEW SCAFFOLD OF THE PHENANTHRIDINONE-BASED INHIBITOR (L10) OF THE SECOND BROMODOMAIN OF BRD2 (BRD2-BD2). HERE, WE PRESENT THE CRYSTAL STRUCTURE OF THE BRD2-BD2, REFINED TO 1.4 A RESOLUTION, IN COMPLEX WITH BETA-MERCAPTOETHANOL (A COMPONENT OF THE PROTEIN BUFFER). THE BETA-MERCAPTOETHANOL COVALENTLY LINKS TO C425 OF BD2 IN THE ACETYL-LYSINE BINDING POCKET, TO FORM A MODIFIED CYSTEINE MERCAPTOETHANOL (CME). THE CME MODIFICATION SIGNIFICANTLY HINDERS THE ENTRY OF LIGANDS INTO THE BD2 BINDING POCKET, SUGGESTING THAT BETA-MERCAPTOETHANOL SHOULD BE REMOVED DURING PROTEIN PRODUCTION PROCESS. NEXT, TO CONFIRM WHETHER PHENANTHRIDIONONE SCAFFOLD IS A NEW INHIBITOR FAMILY OF BRD2-BD2, WE HAVE DETERMINED THE CRYSTAL STRUCTURE OF BD2 IN COMPLEX WITH 6(5H)-PHENANTHRIDINONE (A CORE MOIETY OF L10), REFINED TO 1.28 A RESOLUTION. IT CONFIRMED THAT THE PHENANTHRIDINONE MOLECULE, UNAMBIGUOUSLY, BINDS TO BD2. MOREOVER, WE PERFORMED MOLECULAR DOCKING AND MOLECULAR DYNAMIC STUDIES ON SELECTED PHENANTHRIDINONE ANALOGS. THE PREDICTED L10 ANALOGS ARE STABLE WITH ESSENTIAL HYDROPHOBIC AND HYDROPHILIC INTERACTIONS WITH BD2 DURING MOLECULAR DYNAMIC SIMULATIONS. WE PROPOSE THAT THE PREDICTED PHENANTHRIDINONE ANALOGS MAY BE POTENTIAL MOLECULES FOR INHIBITING THE BD2 FUNCTION OF ACETYLATED HISTONE RECOGNITION. 2018 2 3471 35 IDENTIFICATION AND CHARACTERIZATIONS OF NOVEL, SELECTIVE HISTONE METHYLTRANSFERASE SET7 INHIBITORS BY SCAFFOLD HOPPING- AND 2D-MOLECULAR FINGERPRINT-BASED SIMILARITY SEARCH. SET7, SERVING AS THE ONLY HISTONE METHYLTRANSFERASE THAT MONOMETHYLATES 'LYS-4' OF HISTONE H3, HAS BEEN PROVED TO FUNCTION AS A KEY REGULATOR IN DIVERSE BIOLOGICAL PROCESSES, SUCH AS CELL PROLIFERATION, TRANSCRIPTIONAL NETWORK REGULATION IN EMBRYONIC STEM CELL, CELL CYCLE CONTROL, PROTEIN STABILITY, HEART MORPHOGENESIS AND DEVELOPMENT. WHAT'S MORE, SET7 IS INVOLVED INTHE PATHOGENESIS OF ALOPECIA AERATE, BREAST CANCER, TUMOR AND CANCER PROGRESSION, ATHEROSCLEROSIS IN HUMAN CAROTID PLAQUES, CHRONIC RENAL DISEASES, DIABETES, OBESITY, OVARIAN CANCER, PROSTATE CANCER, HEPATOCELLULAR CARCINOMA, AND PULMONARY FIBROSIS. THEREFORE, THERE IS URGENT NEED TO DEVELOP NOVEL SET7 INHIBITORS. IN THIS PAPER, BASED ON DC-S239 WHICH HAS BEEN PREVIOUSLY REPORTED IN OUR GROUP, WE EMPLOYED SCAFFOLD HOPPING- AND 2D FINGERPRINT-BASED SIMILARITY SEARCHES AND IDENTIFIED DC-S285 AS THE NEW HIT COMPOUND TARGETING SET7 (IC(50) = 9.3 MUM). BOTH RADIOACTIVE TRACING AND NMR EXPERIMENTS VALIDATED THE INTERACTIONS BETWEEN DC-S285 AND SET7 FOLLOWED BY THE SECOND-ROUND SIMILARITY SEARCH LEADING TO THE IDENTIFICATION OFDC-S303 WITH THE IC(50) VALUE OF 1.1 MUM. IN CELLULAR LEVEL, DC-S285 RETARDED TUMOR CELL PROLIFERATION AND SHOWED SELECTIVITY AGAINST MCF7 (IC(50) = 21.4 MUM), JURKAT (IC(50) = 2.2 MUM), THP1 (IC(50) = 3.5 MUM), U937 (IC(50) = 3.9 MUM) CELL LINES. DOCKING CALCULATIONS SUGGESTED THAT DC-S303 SHARE SIMILAR BINDING MODE WITH THE PARENT COMPOUNDDC-S239. WHAT'S MORE, IT PRESENTED GOOD SELECTIVITY AGAINST OTHER EPIGENETIC TARGETS, INCLUDING SETD1B, SETD8, G9A, SMYD2 AND EZH2. DC-S303 CAN SERVE AS A DRUG-LIKE SCAFFOLD WHICH MAY NEED FURTHER OPTIMIZATION FOR DRUG DEVELOPMENT, AND CAN BE USED AS CHEMICAL PROBE TO HELP THE COMMUNITY TO BETTER UNDERSTAND THE SET7 BIOLOGY. 2018 3 1338 32 DESIGN, SYNTHESIS AND BIOLOGICAL EVALUATION OF A PHENYL BUTYRIC ACID DERIVATIVE, N-(4-CHLOROPHENYL)-4-PHENYLBUTANAMIDE: A HDAC6 INHIBITOR WITH ANTI-PROLIFERATIVE ACTIVITY ON CERVIX CANCER AND LEUKEMIA CELLS. BACKGROUND: THE EPIGENETIC REGULATION OF GENES IN CANCER COULD BE TARGETED BY INHIBITING HISTONE DEACETYLASE 6 (HDAC6), AN ENZYME INVOLVED IN SEVERAL TYPES OF CANCER SUCH AS LYMPHOMA, LEUKEMIA, OVARIAN CANCER, ETC. OBJECTIVE: THROUGH IN SILICO METHODS, A SET OF PHENYL BUTYRIC ACID DERIVATIVES WITH POSSIBLE HDAC6 INHIBITORY ACTIVITY WERE DESIGNED, RENDERING MONOPHENYLAMIDES AND BIPHENYLAMIDES USING TUBACIN (HDAC6 SELECTIVE INHIBITOR) AS REFERENCE. METHOD: THE TARGET COMPOUNDS WERE SUBMITTED TO THEORETICAL ADMET ANALYSES AND THEIR BINDING PROPERTIES ON DIFFERENT HDAC6 CONFORMERS WERE EVALUATED THROUGH DOCKING CALCULATIONS. RESULTS: THESE IN SILICO STUDIES ALLOWED US TO IDENTIFY A COMPOUND NAMED B-R2B. IN ORDER TO HAVE MORE INFORMATION ABOUT THE B-R2B BINDING RECOGNITION PROPERTIES ON HDAC6, THE B-R2B-HDAC6 COMPLEX WAS SUBMITTED THROUGH 100 NS-LONG MOLECULAR DYNAMICS (MD) SIMULATION COUPLED TO MMGBSA APPROACH, REVEALING THAT B-R2B IS LOCATED AT THE ENTRANCE OF HDAC6 ACTIVE POCKET, BLOCKING THE PASSAGE OF THE SUBSTRATE WITHOUT REACHING THE HDAC6 BINDING SITE. BASED ON THESE RESULTS, B-R2B WAS SYNTHESIZED, CHARACTERIZED AND BIOLOGICALLY TESTED. THE HDAC6 FLUOROMETRIC DRUG DISCOVERY KIT FLUOR-DE-LYS (ENZO LIFE SCIENCES INC.) WAS USED TO DETERMINE THE HDAC6 HUMAN INHIBITORY ACTIVITY (IC50 VALUE) OF B-R2B COMPOUND. IN ADDITION, B-R2B SHOW IC50 VALUES ON CANCER CELL LINES (HELA; IC50 = 72.6 MICROM), ACUTE MYELOID LEUKEMIA (THP-1; IC50 = 16.5 MICROM), HUMAN MAST LEUKEMIA (HMC; IC50 = 79.29 MICROM) AND CHRONIC MYELOGENOUS LEUKEMIA (KASUMI; IC50 = 101 MICROM). CONCLUSION: THESE RESULTS SHOW THAT B-R2B IS A HDAC6 INHIBITOR, SPECIFICALLY A NON-COMPETITIVE TYPE IN A SIMILAR WAY THAT TUBACIN DOES, ACCORDING TO MD SIMULATIONS. 2017 4 5944 32 TARGETING SMALL MOLECULE TYROSINE KINASES BY POLYPHENOLS: NEW MOVE TOWARDS ANTI-TUMOR DRUG DISCOVERY. BACKGROUND: CANCER IS A COMPLEX DISEASE INVOLVING GENETIC AND EPIGENETIC ALTERATION THAT ALLOWS CELLS TO ESCAPE NORMAL HOMEOSTASIS. KINASES PLAY A CRUCIAL ROLE IN SIGNALING PATHWAYS THAT REGULATE CELL FUNCTIONS. DEREGULATION OF KINASES LEADS TO A VARIETY OF PATHOLOGICAL CHANGES, ACTIVATING CANCER CELL PROLIFERATION AND METASTASES. THE MOLECULAR MECHANISM OF CANCER IS COMPLEX AND THE DYSREGULATION OF TYROSINE KINASES LIKE ANAPLASTIC LYMPHOMA KINASE (ALK), BCR-ABL (FUSION GENE FOUND IN PATIENT WITH CHRONIC MYELOGENOUS LEUKEMIA (CML), JAK (JANUS ACTIVATED KINASE), SRC FAMILY KINASES (SFKS), ALK (ANAPLASTIC LYMPHOMA KINASE), C-MET (MESENCHYMAL- EPITHELIAL TRANSITION), EGFR (EPIDERMAL GROWTH FACTOR RECEPTOR), PDGFR (PLATELET-DERIVED GROWTH FACTOR RECEPTOR), RET (REARRANGED DURING TRANSFECTION) AND VEGFR (VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTOR) PLAYS MAJOR ROLE IN THE PROCESS OF CARCINOGENESIS. RECENTLY, KINASE INHIBITORS HAVE OVERCOME MANY PROBLEMS OF TRADITIONAL CANCER CHEMOTHERAPY AS THEY EFFECTIVELY SEPARATE OUT NORMAL, NON-CANCER CELLS AS WELL AS RAPIDLY MULTIPLYING CANCER CELLS. METHODS: ELECTRONIC DATABASES WERE SEARCHED TO EXPLORE THE SMALL MOLECULE TYROSINE KINASES BY POLYPHENOLS WITH THE HELP OF DOCKING STUDY (GLIDE-7.6 PROGRAM INTERFACED WITH MAESTRO-V11.3 OF SCHRODINGER 2017) TO SHOW THE BINDING ENERGIES OF POLYPHENOLS INHIBITOR WITH DIFFERENT TYROSINE KINASES IN ORDER TO DIFFERENTIATE BETWEEN THE TARGETS. RESULTS: FROM THE LITERATURE SURVEY, IT WAS OBSERVED THAT THE NUMBER OF POLYPHENOLS DERIVED FROM NATURAL SOURCES ALTERS THE EXPRESSION AND SIGNALING CASCADE OF TYROSINE KINASE IN VARIOUS TUMOR MODELS. THEREFORE, THE DEVELOPMENT OF POLYPHENOLS AS A TYROSINE KINASE INHIBITOR AGAINST TARGETED PROTEINS IS REGARDED AS AN UPCOMING TREND FOR CHEMOPREVENTION. CONCLUSION: IN THIS REVIEW, WE HAVE DISCUSSED THE ROLE OF POLYPHENOLS AS CHEMORECEPTIVE WHICH WILL HELP IN FUTURE FOR THE DEVELOPMENT AND DISCOVERY OF NOVEL SEMISYNTHETIC ANTICANCER AGENTS COUPLED WITH POLYPHENOLS. 2020 5 4303 27 MICRORNA-223 INHIBITS TISSUE FACTOR EXPRESSION IN VASCULAR ENDOTHELIAL CELLS. OBJECTIVE: ATHEROSCLEROSIS IS A CHRONIC INFLAMMATORY PROCESS, IN WHICH VASCULAR ENDOTHELIAL CELLS (ECS) BECOME DYSFUNCTIONAL OWING TO THE EFFECTS OF CHEMICAL SUBSTANCES, SUCH AS INFLAMMATORY FACTOR AND GROWTH FACTORS. TISSUE FACTOR (TF) EXPRESSION IS INDUCED BY THE ABOVE CHEMICAL SUBSTANCES IN ACTIVATED ECS. TF INITIATES THROMBOSIS ON DISRUPTED ATHEROSCLEROTIC PLAQUES WHICH PLAYS AN ESSENTIAL ROLE DURING THE ONSET OF ACUTE CORONARY SYNDROMES (ACS). INCREASING EVIDENCES SUGGEST THE IMPORTANT ROLE OF MICRORNAS AS EPIGENETIC REGULATORS OF ATHEROSCLEROTIC DISEASE. THE AIM OF OUR STUDY IS TO IDENTIFY IF MICRORNA-223 (MIR-223) TARGETS TF IN ECS. METHODS AND RESULTS: BIOINFORMATIC ANALYSIS SHOWED THAT TF IS A TARGET CANDIDATE OF MIR-223. WESTERN BLOTTING ANALYSIS REVEALED THAT TUMOR NECROSIS FACTOR ALPHA (TNF-ALPHA) INCREASED TF EXPRESSION IN AORTA OF C57BL/6J MICE AND CULTURED ECS (EA.HY926 CELLS AND HUVEC) AFTER 4 H TREATMENT. IN TNF-ALPHA TREATED ECS, TF MRNA WAS ALSO INCREASED MEASURED BY REAL-TIME PCR. REAL-TIME PCR RESULTS SHOWED THAT MIR-223 LEVELS WERE DOWNREGULATED IN TNF-ALPHA-TREATED AORTA OF C57BL/6J MICE AND CULTURED ECS. TRANSFECTION OF ECS WITH MIR-223 MIMIC OR MIR-223 INHIBITOR MODIFIED TF EXPRESSION BOTH IN MRNA AND PROTEIN LEVELS. LUCIFERASE ASSAYS CONFIRMED THAT MIR-223 SUPPRESSED TF EXPRESSION BY BINDING TO THE SEQUENCE OF TF 3'-UNTRANSLATED REGIONS (3'UTR). TF PROCOAGULANT ACTIVITY WAS INHIBITED BY OVEREXPRESSING MIR-223 WITH OR WITHOUT TNF-ALPHA STIMULATION. CONCLUSIONS: MIR-223-MEDIATED SUPPRESSION OF TF EXPRESSION PROVIDES A NOVEL MOLECULAR MECHANISM FOR THE REGULATION OF COAGULATION CASCADE, AND SUGGESTS A CLUE AGAINST THROMBOGENESIS DURING THE PROCESS OF ATHEROSCLEROTIC PLAQUE RUPTURE. 2014 6 2326 21 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 7 2729 33 EXPLORING EPIGENETIC DRUGS AS POTENTIAL INHIBITORS OF SARS-COV-2 MAIN PROTEASE: A DOCKING AND MD SIMULATION STUDY. THE COVID-19 PANDEMIC HAS CAUSED HAVOC AROUND THE GLOBE SINCE 2019 AND IS CONSIDERED THE LARGEST GLOBAL EPIDEMIC OF THE TWENTIETH CENTURY. ALTHOUGH THE FIRST ANTIVIRAL DRUG, REMDESIVIR, WAS INITIALLY INTRODUCED AGAINST COVID?19, VIRTUALLY NO TANGIBLE THERAPEUTIC DRUGS EXIST TO TREAT SARS-COV-2 INFECTION. FDA-APPROVED PAXLOVID (NIRMATRELVIR SUPPLEMENTED BY RITONAVIR) WAS RECENTLY ANNOUNCED AS A PROMISING DRUG AGAINST THE SARS-COV-2 MAJOR PROTEASE (M(PRO)). HERE WE REPORT FOR THE FIRST TIME THE REMARKABLE INHIBITORY POTENTIALS OF LEAD EPIGENETIC-TARGETING DRUGS (EPI-DRUGS) AGAINST SARS-COV-2 M(PRO). EPI-DRUGS ARE PROMISING COMPOUNDS TO BE USED IN COMBINATION WITH CANCER CHEMOTHERAPEUTICS TO REGULATE GENE EXPRESSION. THE SEARCH FOR ALL KNOWN EPI-DRUGS FOR THE SPECIFIC INHIBITION OF SARS-COV-2 M(PRO) WAS PERFORMED FOR THE FIRST TIME BY CONSENSUS (THREE HIGH-ORDER PROGRAM) MOLECULAR DOCKING STUDIES AND END-STATE FREE ENERGY CALCULATIONS. SEVERAL EPI-DRUGS WERE IDENTIFIED WITH HIGHLY COMPARABLE BINDING AFFINITY TO SARS-COV-2 M(PRO) COMPARED TO NIRMATRELVIR. IN PARTICULAR, POTENT HISTONE METHYLTRANSFERASE INHIBITOR EPZ005687 AND DNA METHYLTRANSFERASE INHIBITOR GUADECITABINE WERE PROMINENT AS THE MOST PROMISING EPI-DRUG INHIBITORS FOR SARS-COV-2 M(PRO). LONG MOLECULAR DYNAMICS (MD) SIMULATIONS (200 NS EACH) AND CORRESPONDING MM-GBSA CALCULATIONS CONFIRMED THE STABILITY OF THE EPZ005687-M(PRO) COMPLEX WITH MM-GBSA BINDING FREE ENERGY (DELTAG(BIND)) -48.2 KCAL/MOL (EPZ005687) COMPARED TO NIRMATRELVIR (-44.7 KCAL/MOL). TAKEN TOGETHER, THE ANTIVIRAL ACTIVITIES OF THE HIGHLIGHTED EPI-DRUGS ARE REPORTED BEYOND WIDESPREAD USE IN COMBINATION WITH ANTI-CANCER AGENTS. THE CURRENT FINDINGS THEREFORE HIGHLIGHT AS YET UNEXPLORED ANTIVIRAL POTENTIAL OF EPI-DRUGS SUITABLE FOR USE IN PATIENTS STRUGGLING WITH CHRONIC IMMUNOSUPPRESSIVE DISORDERS.COMMUNICATED BY RAMASWAMY H. SARMA. 2023 8 5947 28 TARGETING THE EPIGENOME: SCREENING BIOACTIVE COMPOUNDS THAT REGULATE HISTONE DEACETYLASE ACTIVITY. SCOPE: NUTRIGENOMICS IS A RAPIDLY EXPANDING FIELD THAT ELUCIDATES THE LINK BETWEEN DIET-GENOME INTERACTIONS. RECENT EVIDENCE DEMONSTRATES THAT REGULATION OF THE EPIGENOME, AND IN PARTICULAR INHIBITION OF HISTONE DEACETYLASES (HDACS), IMPACT PATHOGENETIC MECHANISMS INVOLVED IN CHRONIC DISEASE. FEW STUDIES, TO DATE, HAVE SCREENED LIBRARIES OF BIOACTIVE COMPOUNDS THAT ACT AS EPIGENETIC MODIFIERS. THIS STUDY SCREENED A LIBRARY OF 131 NATURAL COMPOUNDS TO DETERMINE BIOACTIVE COMPOUNDS THAT INHIBIT ZN-DEPENDENT HDAC ACTIVITY. METHODS AND RESULTS: USING CLASS-SPECIFIC HDAC SUBSTRATES, WE SCREENED 131 NATURAL COMPOUNDS FOR HDAC ACTIVITY IN BOVINE CARDIAC TISSUE. FROM THIS SCREEN, WE IDENTIFIED 18 BIOACTIVE COMPOUND HDAC INHIBITORS. USING OUR CLASS-SPECIFIC HDAC SUBSTRATES, WE NEXT SCREENED THESE 18 BIOACTIVE COMPOUNDS AGAINST RECOMBINANT HDAC PROTEINS. CONSISTENT WITH INHIBITION OF HDAC ACTIVITY, THESE COMPOUNDS WERE CAPABLE OF INHIBITING ACTIVITY OF INDIVIDUAL HDAC ISOFORMS. LASTLY, WE REPORT THAT TREATMENT OF H9C2 CARDIAC MYOBLASTS WITH BIOACTIVE HDAC INHIBITORS WAS SUFFICIENT TO INCREASE LYSINE ACETYLATION AS ASSESSED VIA IMMUNOBLOT. CONCLUSION: THIS STUDY PROVIDED THE FIRST STEP IN IDENTIFYING MULTIPLE BIOACTIVE COMPOUND HDAC INHIBITORS. TAKEN TOGETHER, THIS REPORT SETS THE STAGE FOR FUTURE EXPLORATION OF THESE BIOACTIVE COMPOUNDS AS EPIGENETIC REGULATORS TO POTENTIALLY AMELIORATE CHRONIC DISEASE. 2017 9 2362 22 EPIGENETIC REGULATION OF SNAP25 PREVENTS PROGRESSIVE GLUTAMATE EXCITOTOXICTY IN HYPOXIC CA3 NEURONS. EXPOSURE TO GLOBAL HYPOXIA AND ISCHEMIA HAS BEEN REPORTED TO CAUSE NEURODEGENERATION IN THE HIPPOCAMPUS WITH CA3 NEURONS. THIS NEURONAL DAMAGE IS PROGRESSIVE DURING THE INITIAL PHASE OF EXPOSURE BUT MAINTAINS A PLATEAU ON PROLONGED EXPOSURE. THE PRESENT STUDY ON SPRAGUE DAWLEY RATS AIMED AT UNDERSTANDING THE UNDERLYING MOLECULAR AND EPIGENETIC MECHANISMS THAT LEAD TO HYPOXIC ADAPTATION OF CA3 NEURONS ON PROLONGED EXPOSURE TO A GLOBAL HYPOXIA. OUR RESULTS SHOW STAGNANCY IN NEURODEGENERATION IN CA3 REGION BEYOND 14 DAYS OF CHRONIC EXPOSURE TO HYPOBARIA SIMULATING AN ALTITUDE OF 25,000 FT. DESPITE INCREASED SYNAPTOSOMAL GLUTAMATE AND HIGHER EXPRESSION OF NR1 SUBUNIT OF NMDA RECEPTORS, WE OBSERVED DECREASE IN POST-SYNAPTIC DENSITY AND ACCUMULATION OF SYNAPTIC VESICLES AT THE PRE-SYNAPTIC TERMINALS. MOLECULAR INVESTIGATIONS INVOLVING WESTERN BLOT AND REAL-TIME PCR SHOWED DURATION-DEPENDENT DECREASE IN THE EXPRESSION OF SNAP-25 RESULTING IN REDUCED VESICULAR DOCKING AND SYNAPTIC REMODELING. CHIP ASSAYS FOR EPIGENETIC FACTORS SHOWED DECREASED EXPRESSION OF H3K9AC AND H3K14AC RESULTING IN SNAP-25 PROMOTER SILENCING DURING PROLONGED HYPOXIA. ADMINISTRATION OF SODIUM BUTYRATE, A NON-SPECIFIC HDAC INHIBITOR, DURING 21 DAYS HYPOXIC EXPOSURE PREVENTED SNAP-25 DOWNREGULATION BUT INCREASED CA3 NEURODEGENERATION. THIS EPIGENETIC REGULATION OF SNAP-25 PROMOTER WAS INDEPENDENT OF INCREASED DNMT3B EXPRESSION AND PROMOTER METHYLATION. OUR FINDINGS PROVIDE A NOVEL INSIGHT INTO EPIGENETIC FACTORS-MEDIATED SYNAPTIC REMODELING TO PREVENT EXCITOTOXIC NEURODEGENERATION ON PROLONGED EXPOSURE TO GLOBAL HYPOBARIC HYPOXIA. 2017 10 439 27 ANTILEUKEMIC ACTIVITY OF VALPROIC ACID IN CHRONIC LYMPHOCYTIC LEUKEMIA B CELLS DEFINED BY MICROARRAY ANALYSIS. EPIGENETIC CODE MODIFICATIONS BY HISTONE DEACETYLASE INHIBITORS HAVE RECENTLY BEEN PROPOSED AS POTENTIAL NEW THERAPIES FOR HEMATOLOGICAL MALIGNANCIES. CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) REMAINS INCURABLE DESPITE THE INTRODUCTION OF NEW TREATMENTS. CLL B CELLS ARE CHARACTERIZED BY AN APOPTOSIS DEFECT RATHER THAN EXCESSIVE PROLIFERATION, BUT PROLIFERATION CENTERS HAVE BEEN FOUND IN ORGANS SUCH AS THE BONE MARROW AND LYMPH NODES. IN THIS STUDY, WE ANALYZED GENE EXPRESSION MODIFICATIONS IN CLL B CELLS AFTER TREATMENT WITH VALPROIC ACID (VPA), A WELL-TOLERATED ANTI-EPILEPTIC DRUG WITH HDAC INHIBITORY ACTIVITY. CLL B CELLS OBTAINED FROM 14 PATIENTS WERE TREATED IN VITRO WITH A CONCENTRATION OF 1 MM VPA FOR 4 H. VPA EFFECTS ON GENE EXPRESSION WERE THEREAFTER STUDIED USING AFFYMETRIX TECHNOLOGY, AND SOME IDENTIFIED GENES WERE VALIDATED BY REAL-TIME PCR AND WESTERN BLOT. WE OBSERVED THAT VPA INDUCED APOPTOSIS BY DOWNREGULATING SEVERAL ANTI-APOPTOTIC GENES AND BY UPREGULATING PRO-APOPTOTIC GENES. FURTHERMORE, VPA SIGNIFICANTLY INCREASED CHEMOSENSITIVITY TO FLUDARABINE, FLAVOPIRIDOL, BORTEZOMIB, THALIDOMIDE AND LENALIDOMIDE. VPA INHIBITED THE PROLIFERATION OF CPG/IL2-STIMULATED CLL B CELLS AND MODULATED MANY CELL CYCLE MESSENGER RNAS. IN CONCLUSION, EXPOSURE OF CLL B CELLS TO VPA INDUCED APOPTOSIS, POTENTIATED CHEMOTHERAPEUTIC AGENT EFFECTS AND INHIBITED PROLIFERATION. THESE DATA STRONGLY SUGGEST THE USE OF VPA IN CLL TREATMENT, PARTICULARLY IN COMBINATION WITH ANTILEUKEMIA AGENTS. 2009 11 2189 25 EPIGENETIC MECHANISMS UNDERLYING THE THERAPEUTIC EFFECTS OF HDAC INHIBITORS IN CHRONIC MYELOID LEUKEMIA. CHRONIC MYELOID LEUKEMIA (CML) IS A HEMATOLOGICAL DISORDER CAUSED BY THE ONCOGENIC BCR-ABL FUSION PROTEIN IN MORE THAN 90% OF PATIENTS. DESPITE THE STRIKING IMPROVEMENTS IN THE MANAGEMENT OF CML PATIENTS SINCE THE INTRODUCTION OF TYROSINE KINASE INHIBITORS (TKIS), THE APPEARANCE OF TKI RESISTANCE AND SIDE EFFECTS LEAD TO TREATMENT FAILURE, JUSTIFYING THE NEED OF NOVEL THERAPEUTIC APPROACHES. HISTONE DEACETYLASE INHIBITORS (HDACIS), ABLE TO MODULATE GENE EXPRESSION PATTERNS AND IMPORTANT CELLULAR SIGNALING PATHWAYS THROUGH THE REGULATION OF THE ACETYLATION STATUS OF BOTH HISTONE AND NON-HISTONE PROTEIN TARGETS, HAVE BEEN REPORTED TO DISPLAY PROMISING ANTI-LEUKEMIC PROPERTIES ALONE OR IN COMBINATION WITH TKIS. THIS REVIEW SUMMARIZES PRE-CLINICAL AND CLINICAL STUDIES THAT INVESTIGATED THE MECHANISMS UNDERLYING THE ANTICANCER POTENTIAL OF HDACIS AND DISCUSSES THE RATIONALE FOR A COMBINATION OF HDACIS WITH TKIS AS A THERAPEUTIC OPTION IN CML. 2020 12 826 25 CHARACTERIZATION OF K562 CELLS: UNCOVERING NOVEL CHROMOSOMES, ASSESSING TRANSFERRIN RECEPTOR EXPRESSION, AND PROBING PHARMACOLOGICAL THERAPIES. HUMAN ERYTHROLEUKEMIC K562 CELLS REPRESENT THE PROTOTYPICAL CELL CULTURE MODEL OF CHRONIC MYELOID LEUKEMIA (CML). THE CELLS ARE PSEUDO-TRIPLOID AND POSITIVE FOR THE PHILADELPHIA CHROMOSOME. THEREFORE, K562 CELLS HAVE BEEN WIDELY USED FOR INVESTIGATING THE BCR/ABL1 ONCOGENE AND THE TYROSINE KINASE INHIBITOR, IMATINIB-MESYLATE. FURTHER, K562 CELLS OVEREXPRESS TRANSFERRIN RECEPTORS (TFR) AND HAVE BEEN USED AS A MODEL FOR TARGETING CYTOTOXIC THERAPIES, VIA RECEPTOR-MEDIATED ENDOCYTOSIS. HERE, WE HAVE CHARACTERIZED K562 CELLS FOCUSING ON THE KARYOTYPE OF CELLS IN PROLONGED CULTURE, REGULATION OF EXPRESSION OF TFR IN WILDTYPE (WT) AND DOXORUBICIN-RESISTANT CELLS, AND RESPONSES TO HISTONE DEACETYLASE INHIBITION (HDACI). KARYOTYPE ANALYSIS INDICATES NOVEL CHROMOSOMES AND GENE EXPRESSION ANALYSIS SUGGESTS A SHIFT OF CULTURED K562 CELLS AWAY FROM PATIENT-DERIVED LEUKEMIC CELLS. WE CONFIRM THE HIGH EXPRESSION OF TFR ON K562 CELLS USING IMMUNOFLUORESCENCE AND CELL-SURFACE RECEPTOR BINDING RADIOASSAYS. IMPORTANTLY, HIGH TFR EXPRESSION IS OBSERVED IN PATIENT-DERIVED CELLS, AND WE HIGHLIGHT THE PERSISTENT EXPRESSION OF TFR FOLLOWING DOXORUBICIN ACQUIRED RESISTANCE. EPIGENETIC ANALYSIS INDICATES THAT PERMISSIVE HISTONE ACETYLATION AND METHYLATION AT THE PROMOTER REGION REGULATES THE TRANSCRIPTION OF TFR IN K562 CELLS. FINALLY, WE SHOW RELATIVELY HIGH EXPRESSION OF HDAC ENZYMES IN K562 CELLS AND DEMONSTRATE THE CHEMOTOXIC EFFECTS OF HDACI, USING THE FDA-APPROVED HYDROXAMIC ACID, VORINOSTAT. TOGETHER WITH A DESCRIPTION OF MORPHOLOGY, INFRARED SPECTRAL ANALYSIS, AND EXAMINATION OF METABOLIC PROPERTIES, WE PROVIDE A COMPREHENSIVE CHARACTERIZATION OF K562 CELLS. OVERALL, K562 CELL CULTURE SYSTEMS REMAIN WIDELY USED FOR THE INVESTIGATION OF NOVEL THERAPEUTICS FOR CML, WHICH IS PARTICULARLY IMPORTANT IN CASES OF IMATINIB-MESYLATE RESISTANCE. 2023 13 1826 25 EFFECTS OF HISTONE DEACETYLASE INHIBITOR ON EXTRACELLULAR MATRIX PRODUCTION IN HUMAN NASAL POLYP ORGAN CULTURES. BACKGROUND: NASAL POLYPOSIS IS ASSOCIATED WITH A CHRONIC INFLAMMATORY CONDITION OF THE SINONASAL MUCOSA AND INVOLVES MYOFIBROBLAST DIFFERENTIATION AND EXTRACELLULAR MATRIX (ECM) ACCUMULATION. EPIGENETIC MODULATION BY HISTONE DEACETYLASE (HDAC) INHIBITORS INCLUDING TRICHOSTATIN A (TSA) HAS BEEN REPORTED TO HAVE INHIBITORY EFFECTS ON MYOFIBROBLAST DIFFERENTIATION IN LUNG AND RENAL FIBROBLASTS. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE THE INHIBITORY EFFECT OF TSA ON MYOFIBROBLAST DIFFERENTIATION AND ECM PRODUCTION IN NASAL POLYP ORGAN CULTURES. METHODS: NASAL POLYP TISSUES FROM 18 PATIENTS WERE ACQUIRED DURING ENDOSCOPIC SINUS SURGERY. AFTER ORGAN CULTURE, NASAL POLYPS WERE STIMULATED WITH TGF-BETA1 AND THEN TREATED WITH TSA. ALPHA-SMOOTH MUSCLE ACTIN (ALPHA-SMA), FIBRONECTIN, AND COLLAGEN TYPE I EXPRESSION LEVELS WERE EXAMINED BY REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION (PCR), REAL-TIME PCR, WESTERN BLOT, AND IMMUNOFLUORESCENT STAINING. HDAC2, HDAC4, AND ACETYLATED H4 EXPRESSION LEVELS WERE ASSAYED BY WESTERN BLOT. CYTOTOXICITY WAS ANALYZED BY THE TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE BIOTIN-DUTP NICK END LABELING ASSAY. RESULTS: THE EXPRESSION LEVELS OF ALPHA-SMA, FIBRONECTIN, AND COLLAGEN TYPE 1 WERE INCREASED IN NASAL POLYP AFTER TRANSFORMING GROWTH FACTOR (TGF) BETA1 TREATMENT. TSA-INHIBITED TGF-BETA1 INDUCED THESE GENE AND PROTEIN EXPRESSION LEVELS. FURTHERMORE, TSA SUPPRESSED PROTEIN EXPRESSION LEVELS OF HDAC2 AND HDAC4. HOWEVER, TSA INDUCED HYPERACETYLATION OF HISTONES H4. TREATMENT WITH TGF-BETA1 WITH OR WITHOUT TSA DID NOT HAVE CYTOTOXIC EFFECT. CONCLUSION: THESE FINDINGS PROVIDE NOVEL INSIGHTS INTO THE EPIGENETIC REGULATION IN MYOFIBROBLAST DIFFERENTIATION AND ECM PRODUCTION OF NASAL POLYP. TSA COULD BE A CANDIDATE OF A THERAPEUTIC AGENT FOR REVERSING THE TGF-BETA1-INDUCED ECM SYNTHESIS THAT LEADS TO NASAL POLYP DEVELOPMENT. 2013 14 1339 32 DESIGN, SYNTHESIS, BIOLOGICAL EVALUATION, AND STRUCTURAL CHARACTERIZATION OF POTENT HISTONE DEACETYLASE INHIBITORS BASED ON CYCLIC ALPHA/BETA-TETRAPEPTIDE ARCHITECTURES. HISTONE DEACETYLASES (HDACS) ARE A FAMILY OF ENZYMES FOUND IN BACTERIA, FUNGI, PLANTS, AND ANIMALS THAT PROFOUNDLY AFFECT CELLULAR FUNCTION BY CATALYZING THE REMOVAL OF ACETYL GROUPS FROM -N-ACETYLATED LYSINE RESIDUES OF VARIOUS PROTEIN SUBSTRATES INCLUDING HISTONES, TRANSCRIPTION FACTORS, ALPHA-TUBULIN, AND NUCLEAR IMPORTERS. ALTHOUGH THE PRECISE ROLES OF HDAC ISOFORMS IN CELLULAR FUNCTION ARE NOT YET COMPLETELY UNDERSTOOD, INHIBITION OF HDAC ACTIVITY HAS EMERGED AS A PROMISING APPROACH FOR REVERSING THE ABERRANT EPIGENETIC STATES ASSOCIATED WITH CANCER AND OTHER CHRONIC DISEASES. POTENT NEW ISOFORM-SELECTIVE HDAC INHIBITORS WOULD THEREFORE HELP EXPAND OUR UNDERSTANDING OF THE HDAC ENZYMES AND REPRESENT ATTRACTIVE LEAD COMPOUNDS FOR DRUG DESIGN, ESPECIALLY IF COMBINED WITH HIGH-RESOLUTION STRUCTURAL ANALYSES OF SUCH INHIBITORS TO SHED LIGHT ON THE THREE-DIMENSIONAL PHARMACOPHORIC FEATURES NECESSARY FOR THE FUTURE DESIGN OF MORE POTENT AND SELECTIVE COMPOUNDS. HERE WE PRESENT STRUCTURAL AND FUNCTIONAL ANALYSES OF A SERIES OF BETA-AMINO-ACID-CONTAINING HDAC INHIBITORS INSPIRED BY CYCLIC TETRAPEPTIDE NATURAL PRODUCTS. TO SURVEY A DIVERSE ENSEMBLE OF PHARMACOPHORIC CONFIGURATIONS, WE SYSTEMATICALLY VARIED THE POSITION OF THE BETA-AMINO ACID, AMINO ACID CHIRALITY, FUNCTIONALIZATION OF THE ZN(2+)-COORDINATING AMINO ACID SIDE CHAIN, AND ALKYLATION OF THE BACKBONE AMIDE NITROGEN ATOMS AROUND THE MACROCYCLE. IN MANY CASES, THE COMPOUNDS WERE A SINGLE CONFORMATION IN SOLUTION AND EXHIBITED POTENT ACTIVITIES AGAINST A NUMBER OF HDAC ISOFORMS AS WELL AS EFFECTIVE ANTIPROLIFERATIVE AND CYTOTOXIC ACTIVITIES AGAINST HUMAN TUMOR CELLS. HIGH-RESOLUTION NMR SOLUTION STRUCTURES WERE DETERMINED FOR A SELECTION OF THE INHIBITORS, PROVIDING A USEFUL MEANS OF CORRELATING DETAILED STRUCTURAL INFORMATION WITH POTENCY. THE STRUCTURE-BASED APPROACH DESCRIBED HERE IS EXPECTED TO FURNISH VALUABLE INSIGHTS TOWARD THE FUTURE DESIGN OF MORE SELECTIVE HDAC INHIBITORS. 2009 15 3191 27 HDAC AND HAT INHIBITORS DIFFERENTLY AFFECT ANALGESIA MEDIATED BY GROUP II METABOTROPIC GLUTAMATE RECEPTORS. BACKGROUND: HISTONE DEACETYLASES (HDACS) AND HISTONE ACETYLTRANSFERASES (HATS) ARE KEY PLAYERS IN EPIGENETIC REGULATION OF GENE EXPRESSION. ANALGESIC ACTIVITY BY HDAC INHIBITORS HAS BEEN REPORTED IN DIFFERENT PAIN MODELS INCLUDING INFLAMMATORY AND NEUROPATHIC PAIN. THESE DRUGS INTERFERE WITH GENE EXPRESSION THROUGH DIFFERENT MECHANISMS INCLUDING CHROMATIN REMODELING AND/OR ACTIVATION OF TRANSCRIPTION FACTORS. AMONG OTHER TARGETS, HDAC INHIBITORS REGULATE METABOTROPIC GLUTAMATE RECEPTORS TYPE 2 (MGLU2) EXPRESSION IN CENTRAL AND PERIPHERAL CENTRAL NERVOUS SYSTEM. HOWEVER WHETHER INHIBITION OF HAT ACTIVITY ALSO REGULATES MGLU2 EXPRESSION HAS NOT BEEN REPORTED. FINDINGS: HERE WE REPORT THAT CURCUMIN (CUR), A NATURALLY OCCURRING COMPOUND ENDOWED WITH P300/CREB-BINDING PROTEIN HAT INHIBITORY ACTIVITY, IS ABLE TO INDUCE A DRASTIC DOWN-REGULATION OF THE MGLU2 RECEPTOR IN THE MOUSE SPINAL CORD AFTER SYSTEMIC ADMINISTRATION TOGETHER WITH A MARKED HYPOACETYLATION OF HISTONES H3 AND H4 IN DORSAL ROOT GANGLIA (DRG). FURTHERMORE, THE ANALGESIC ACTIVITY OF THE MGLU2/3 AGONIST, LY379268 IS LOST AFTER A 3-DAY TREATMENT WITH CUR. CONVERSELY THE ANALGESIC ACTIVITY OF LY379268 IS POTENTIATED IN MICE PRETREATED FOR 5 CONSECUTIVE DAYS WITH THE HDAC INHIBITOR, SUBEROYLANILIDE HYDROXAMIC ACID (SAHA), KNOWN TO INDUCE MGLU2-UPREGULATION. CONCLUSIONS: OUR RESULTS DEMONSTRATE THAT SYSTEMICALLY INJECTED CUR IS ABLE TO INHIBIT H3 AND H4 ACETYLATION IN THE DRG AND TO DOWN-REGULATE MGLU2 RECEPTORS IN THE SPINAL CORD. WE ALSO DEMONSTRATE THAT LONG TERM MODIFICATION OF THE MGLU2 EXPRESSION AFFECTS THE ANALGESIC PROPERTIES OF THE ORTHOSTERIC MGLU2/3 AGONIST, LY379268. THESE DATA OPEN UP THE POSSIBILITY THAT EPIGENETIC MODULATORS MIGHT BE GIVEN IN COMBINATION WITH "TRADITIONAL" DRUGS IN A CONTEXT OF A MULTI TARGET APPROACH FOR A BETTER ANALGESIC EFFICACY. 2014 16 1046 25 CLINICAL DEVELOPMENT OF DECITABINE AS A PROTOTYPE FOR AN EPIGENETIC DRUG PROGRAM. THIS REVIEW HIGHLIGHTS DECITABINE AS A PROTOTYPE EPIGENETIC MODIFYING DRUG TO SHOW HOW THE CLINICAL DEVELOPMENT OF EPIGENETIC AGENTS DIFFERS FROM THAT OF TRADITIONAL CYTOTOXIC CHEMOTHERAPIES. DECITABINE, A CYTOSINE ANALOGUE, IS CYTOTOXIC AT HIGH DOSES BUT HAS SELECTIVE DNA DEMETHYLATING ACTIVITY AT LOW DOSES. THE FOCUS OF CURRENT DECITABINE INVESTIGATIONS IS TWOFOLD: TO ELUCIDATE ALL OF THE MECHANISMS OF ACTION AND TO DETERMINE THE OPTIMAL DOSE, SCHEDULE, AND CONCOMITANT THERAPIES. NEW PHASE I TRIALS HAVE IDENTIFIED A "BIOLOGICALLY EFFECTIVE DOSE," WHICH IS 1 TO 2 LOGS LOWER THAN THE CYTOTOXIC DOSE. A CLINICAL DEVELOPMENT PROGRAM WITH LOW-DOSE DECITABINE IN MALIGNANT DISEASES IS FOCUSED ON MYELODYSPLASTIC SYNDROME (MDS), ACUTE MYELOGENOUS LEUKEMIA (AML), AND CHRONIC MYELOGENOUS LEUKEMIA (CML). A PHASE III TRIAL IN MDS SHOWED OBJECTIVE RESPONSES (COMPLETE [CR] PLUS PARTIAL [PR] REMISSION) AND LONGER MEDIAN TIME TO PROGRESSION TO AML OR DEATH WITH DECITABINE THAN WITH SUPPORTIVE CARE ALONE. THE OPTIMAL USE OF DECITABINE MAY BE IN COMBINATION WITH OTHER AGENTS THAT PROMOTE GENE EXPRESSION, NAMELY, HISTONE DEACETYLASE (HDAC) INHIBITORS. OPTIMIZED DECITABINE DOSES AND COMBINATIONS WITH OTHER EPIGENETIC THERAPIES THAT CAN BE USED AT MINIMALLY TOXIC DOSES PROVIDE POTENTIALLY SAFER THERAPEUTIC OPTIONS AND INTRODUCE NOVEL COMBINATION THERAPIES. 2005 17 881 25 CHRONIC CLOZAPINE TREATMENT RESTRAINS VIA HDAC2 THE PERFORMANCE OF MGLU2 RECEPTOR AGONISM IN A RODENT MODEL OF ANTIPSYCHOTIC ACTIVITY. PRECLINICAL FINDINGS IN RODENT MODELS POINTED TOWARD ACTIVATION OF METABOTROPIC GLUTAMATE 2/3 (MGLU2/3) RECEPTORS AS A NEW PHARMACOLOGICAL APPROACH TO TREAT PSYCHOSIS. HOWEVER, MORE RECENT STUDIES FAILED TO SHOW CLINICAL EFFICACY OF MGLU2/3 RECEPTOR AGONISM IN SCHIZOPHRENIA PATIENTS. WE PREVIOUSLY PROPOSED THAT LONG-TERM ANTIPSYCHOTIC MEDICATION RESTRICTED THE THERAPEUTIC EFFECTS OF THESE GLUTAMATERGIC AGENTS. HOWEVER, LITTLE IS KNOWN ABOUT THE MOLECULAR MECHANISM UNDERLYING THE POTENTIAL REPERCUSSION OF PREVIOUS ANTIPSYCHOTIC EXPOSURE ON THE THERAPEUTIC PERFORMANCE OF MGLU2/3 RECEPTOR AGONISTS. HERE WE SHOW THAT THIS MALADAPTIVE EFFECT OF ANTIPSYCHOTIC TREATMENT IS MEDIATED MOSTLY VIA HISTONE DEACETYLASE 2 (HDAC2). CHRONIC TREATMENT WITH THE ANTIPSYCHOTIC CLOZAPINE LED TO A DECREASE IN MOUSE FRONTAL CORTEX MGLU2 MRNA, AN EFFECT THAT REQUIRED EXPRESSION OF BOTH HDAC2 AND THE SEROTONIN 5-HT(2A) RECEPTOR. THIS TRANSCRIPTIONAL ALTERATION OCCURRED IN ASSOCIATION WITH HDAC2-DEPENDENT REPRESSIVE HISTONE MODIFICATIONS AT THE MGLU2 PROMOTER. WE FOUND THAT CHRONIC CLOZAPINE TREATMENT DECREASED VIA HDAC2 THE CAPABILITIES OF THE MGLU2/3 RECEPTOR AGONIST LY379268 TO ACTIVATE G-PROTEINS IN THE FRONTAL CORTEX OF MICE. CHRONIC CLOZAPINE TREATMENT BLUNTED THE ANTIPSYCHOTIC-RELATED BEHAVIORAL EFFECTS OF LY379268, AN EFFECT THAT WAS NOT OBSERVED IN HDAC2 KNOCKOUT MICE. MORE IMPORTANTLY, CO-ADMINISTRATION OF THE CLASS I AND II HDAC INHIBITOR SAHA (VORINOSTAT) PRESERVED THE ANTIPSYCHOTIC PROFILE OF LY379268 AND FRONTAL CORTEX MGLU2/3 RECEPTOR DENSITY IN WILD-TYPE MICE. THESE FINDINGS RAISE CONCERNS ON THE DESIGN OF PREVIOUS CLINICAL STUDIES WITH MGLU2/3 AGONISTS, PROVIDING THE RATIONALE FOR THE DEVELOPMENT OF HDAC2 INHIBITORS AS A NEW EPIGENETIC-BASED APPROACH TO IMPROVE THE CURRENTLY LIMITED RESPONSE TO TREATMENT WITH GLUTAMATERGIC ANTIPSYCHOTICS. 2019 18 5132 33 POTENTIAL EFFECT OF LUTEOLIN, EPIAFZELECHIN, AND ALBIGENIN ON RATS UNDER CADMIUM-INDUCED INFLAMMATORY INSULT: IN SILICO AND IN VIVO APPROACH. INTRODUCTION: CADMIUM(CD) AN INDUSTRIAL POISON PRESENT ABUNDANTLY IN THE ENVIRONMENT, CAUSES HUMAN TOXICITY BY AN INFLAMMATORY PROCESS. CHRONIC EXPOSURE OF CADMIUM CAN CAUSE A NUMBER OF MOLECULAR LESIONS THAT COULD BE RELEVANT TO ONCOGENESIS, THROUGH INDIRECT OR EPIGENETIC MECHANISMS, POTENTIALLY INCLUDING ABNORMAL ACTIVATION OF ONCOGENES AND SUPPRESSION OF APOPTOSIS BY DEPLETION OF ANTIOXIDANTS. AS INDUCTION OF CYCLOOXYGENASE (COX)-2 IS LINKED TO INFLAMMATORY PROCESSES, USE OF LUTEOLIN, EPIAFZELECHIN, AND ALBIGENIN ALONE OR IN DIFFERENT COMBINATIONS MAY BE USED AS ANTI-INFLAMMATORY THERAPEUTIC AGENTS. METHODS: WE, HEREIN, PERFORMED IN SILICO EXPERIMENTS TO CHECK THE BINDING AFFINITY OF PHYTOCHEMICALS AND THEIR THERAPEUTIC EFFECT AGAINST COX-2 IN CADMIUM ADMINISTERED RATS. WISTAR ALBINO RATS WERE GIVEN PHYTOCHEMICALS IN DIFFERENT COMBINATIONS TO CHECK THEIR ANTI-INFLAMMATORY ACTIVITIES AGAINST CADMIUM INTOXICATION. THE LEVEL OF ALANINE AMINOTRANSFERASES (ALT), 4-HYDROXYNONENAL (4HNE), 8-HYDROXY-2-DEOXYGUANOSINE (8-OHDG), TUMOR NECROSIS FACTOR-ALPHA (TNF-ALPHA), ISOPROSTANES (ISOP-2ALPHA), COX-2, AND MALONDIALDEHYDE (MDA) WERE ESTIMATED WITH THEIR RESPECTIVE ELISA AND SPECTROPHOTOMETRIC METHODS. RESULTS: THE GENERATED RESULTS SHOW THAT PHYTOCOMPOUNDS POSSESSED GOOD BINDING ENERGY POTENTIAL AGAINST COX-2, AND COMMON INTERACTIVE BEHAVIOR WAS OBSERVED IN ALL DOCKING STUDIES. MOREOVER, THE LEVEL OF ALT, 4HNE, 8-OHDG, TNF-ALPHA, ISOP-2ALPHA, MALONDIALDEHYDE, AND COX-2 WERE SIGNIFICANTLY INCREASED IN RATS WITH INDUCED TOXICITY COMPARED TO THE CONTROL GROUP, WHEREAS IN COMBINATIONAL THERAPY OF PHYTOCOMPOUNDS, THE LEVELS WERE SIGNIFICANTLY DECREASED IN THE GROUP. DISCUSSION: TAKEN TOGETHER, LUTEOLIN, EPIAFZELECHIN, AND ALBIGENIN CAN BE USED AS ANTI-INFLAMMATORY THERAPEUTIC AGENTS FOR FUTURE NOVEL DRUG DESIGN, AND THUS IT MAY HAVE THERAPEUTIC IMPORTANCE AGAINST CADMIUM TOXICITY. 2023 19 2437 23 EPIGENETIC SILENCING OF SONIC HEDGEHOG ELICITS ANTITUMOR IMMUNE RESPONSE AND SUPPRESSES TUMOR GROWTH BY INHIBITING THE HEDGEHOG SIGNALING PATHWAY IN METASTATIC SPINE TUMORS IN SPRAGUE-DAWLEY RATS. BACKGROUND: PATIENTS WITH METASTATIC SPINE TUMORS MAY SUFFER FROM PAIN OR NEUROLOGIC DEFICIT, AND THE DISEASE MAY BE DETECTED IN PATIENTS WITH A KNOWN MALIGNANCY. SONIC HEDGEHOG (SHH) HAS RECEIVED SPECIAL ATTENTION DUE TO ITS ROLE IN CANCERS. THEREFORE, THIS STUDY INVESTIGATED THE EFFECTS OF EPIGENETIC SILENCING OF SHH ON ANTITUMOR IMMUNE RESPONSE AND TUMOR GROWTH BY REGULATING THE HEDGEHOG (HH) SIGNALING PATHWAY IN METASTATIC SPINE TUMORS. METHODS: RAT MODELS OF METASTATIC SPINE TUMORS WERE SUCCESSFULLY ESTABLISHED. WE FIRST CALCULATED THE TUMOR VOLUME AND THE INHIBITION RATE OF TUMOR GROWTH TO INVESTIGATE THE EFFECT OF SHH ON TUMOR GROWTH. AFTERWARDS, IMMUNOHISTOCHEMISTRY WAS USED TO DETERMINE WHETHER PROLIFERATION WAS DELAYED BY SHH DEPLETION, AND THE 3-(4,5-DIMETHYLTHIAZOL-2-YL)-2,5-DIPHENYLTETRAZOLIUM BROMIDE ASSAY WAS CONDUCTED TO TEST THE CHANGES IN THE LYMPHOCYTE TRANSFORMATION RATE IN THE SPLEEN TRIGGERED BY SHH SILENCING. THEN, THE INFLUENCE OF SHH DEPLETION ON IMMUNE FUNCTION WAS INVESTIGATED. LATER, QUANTITATIVE REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION AND WESTERN BLOT ASSAY WERE PERFORMED TO EXPLORE THE HH SIGNALING PATHWAY-RELATED FACTORS. FINALLY, WE ADDED THE HH SIGNALING PATHWAY INHIBITOR, GDC-0449, TO CONFIRM THE ROLE OF THE PATHWAY IN TUMOR PROGRESSION. RESULTS: INITIALLY, WE OBSERVED THAT SHH DEPLETION WAS A NEGATIVE FACTOR FOR TUMOR GROWTH. AFTERWARDS, IT WAS REVEALED THAT EPIGENETIC SILENCING OF SHH SERVED AS AN INHIBITOR FACTOR FOR THE FUNCTION OF SPLEEN LYMPHOCYTE TRANSFORMATION AND INFLAMMATION WHILE PROMOTING ANTITUMOR IMMUNE FUNCTION. CONCLUSION: OUR PRELIMINARY RESULTS INDICATE THAT EPIGENETIC SILENCING OF SHH ELICITS AN ANTITUMOR IMMUNE RESPONSE AND SUPPRESSES TUMOR GROWTH BY INHIBITING THE HH SIGNALING PATHWAY IN METASTATIC SPINE TUMORS. 2018 20 1105 32 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