1 1338 160 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 2 950 23 CHRONIC MILD STRESS EXACERBATES SEVERITY OF EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS IN ASSOCIATION WITH ALTERED NON-CODING RNA AND METABOLIC BIOMARKERS. THE CAUSAL FACTORS DETERMINING THE ONSET AND SEVERITY OF MULTIPLE SCLEROSIS (MS) ARE NOT WELL UNDERSTOOD. HERE, WE INVESTIGATED THE INFLUENCE OF CHRONIC STRESS ON CLINICAL SYMPTOMS, METABOLIC AND EPIGENETIC MANIFESTATIONS OF EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE), A COMMON ANIMAL MODEL OF MS. LEWIS RATS WERE IMMUNIZED FOR MONOPHASIC EAE WITH MBP(69-88) AND WERE EXPOSED TO CHRONIC STRESS FOR 37DAYS STARTING 7DAYS PRIOR TO IMMUNIZATION. THE EXPOSURE TO STRESS ACCELERATED AND EXACERBATED THE CLINICAL SYMPTOMS OF EAE. BOTH STRESS AND EAE ALSO DISRUPTED METABOLIC STATUS AS INDICATED BY TRACE ELEMENTAL ANALYSIS IN BODY HAIR. STRESS PARTICULARLY EXACERBATED CHLORINE DEPOSITION IN EAE ANIMALS. MOREOVER, DEEP SEQUENCING REVEALED A CONSIDERABLE IMPACT OF STRESS ON MICRORNA EXPRESSION IN EAE. EAE BY ITSELF UPREGULATED MICRORNA EXPRESSION IN LUMBAR SPINAL CORD, INCLUDING MIR-21, MIR-142-3P, MIR-142-5P, MIR-146A, AND MIR-155. STRESS IN EAE FURTHER UP-REGULATED MIR-16, MIR-146A AND MIR-155 LEVELS. THE LATTER TWO MICRORNAS ARE RECOGNIZED BIOMARKERS OF HUMAN MS. THUS, STRESS MAY SYNERGISTICALLY EXACERBATE SEVERITY OF EAE BY ALTERING EPIGENETIC REGULATORY PATHWAYS. THE FINDINGS SUGGEST THAT STRESS MAY REPRESENT A SIGNIFICANT RISK FACTOR FOR SYMPTOMATIC DETERIORATION IN MS. STRESS-RELATED METABOLIC AND MICRORNA SIGNATURES SUPPORT THEIR VALUE AS BIOMARKERS FOR PREDICTING THE RISK AND SEVERITY OF MS. 2017 3 3347 26 HISTONE DEACETYLASES MEDIATE THE SILENCING OF MIR-15A, MIR-16, AND MIR-29B IN CHRONIC LYMPHOCYTIC LEUKEMIA. CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) DEMONSTRATES A GLOBAL DOWN-REGULATION OF MIR-15A AND MIR-16 AND A SELECTIVE SILENCING OF THE RELATED MIR-29B IN AGGRESSIVE DISEASE. DELETIONS IN CHROMOSOME 13 [DEL(13Q14)] PARTIALLY ACCOUNT FOR THE LOSS OF EXPRESSION OF MIR-15A AND MIR-16, BUT THE MECHANISMS BY WHICH MIR-29B BECOMES SILENCED IS UNKNOWN. IN THE PRESENT STUDY, WE SHOW THAT THE HISTONE DEACETYLASES (HDACS) ARE OVEREXPRESSED IN CLL AND MEDIATE THE EPIGENETIC SILENCING OF MIR-15A, MIR-16, AND MIR-29B. HDAC INHIBITION TRIGGERED THE ACCUMULATION OF THE TRANSCRIPTIONALLY ACTIVATING CHROMATIN MODIFICATION H3K4ME2 AND RESTORED THE EXPRESSION OF MIR-15A, MIR-16, AND MIR-29B IN APPROXIMATELY 35% OF SAMPLES. ECTOPIC EXPRESSION OF MIR-15A AND MIR-16 AND HDAC INHIBITION-INDUCED EXPRESSION OF MIR-15A, MIR-16, OR MIR-29B IN PRIMARY CLL CELLS WAS ASSOCIATED WITH DECLINES IN THE LEVELS OF MCL-1, BUT NOT BCL-2, MITOCHONDRIAL DYSFUNCTION, AND INDUCTION OF CELL DEATH. THEREFORE, OUR RESULTS SHOW THAT HDACS ABERRANTLY SILENCE THE EXPRESSION OF THE CRITICAL TUMOR SUPPRESSORS MIR-15A, MIR-16, AND MIR-29B IN CLL. DEACETYLASE INHIBITION MAY BE A THERAPEUTIC STRATEGY THAT RESTORES THE EXPRESSION OF THESE MIRS TO ANTAGONIZE MCL-1, AN IMPORTANT SURVIVAL PROTEIN IN THESE CELLS. CONSEQUENTLY, CLL PATIENTS WHO EXHIBIT SUCH EPIGENETIC SILENCING MAY BENEFIT FROM HDAC INHIBITOR-BASED THERAPY. 2012 4 6586 32 TUBASTATIN, A SELECTIVE HISTONE DEACETYLASE 6 INHIBITOR SHOWS ANTI-INFLAMMATORY AND ANTI-RHEUMATIC EFFECTS. EPIGENETIC MODIFICATIONS REPRESENT A PROMISING NEW APPROACH TO MODULATE CELL FUNCTIONS AS OBSERVED IN AUTOIMMUNE DISEASES. EMERGING EVIDENCE SUGGESTS THE UTILITY OF HDAC INHIBITORS IN THE TREATMENT OF CHRONIC IMMUNE AND INFLAMMATORY DISORDERS. HOWEVER, CLASS AND ISOFORM SELECTIVE INHIBITION OF HDAC IS CURRENTLY FAVORED AS IT LIMITS THE TOXICITY THAT HAS BEEN OBSERVED WITH PAN-HDAC INHIBITORS. HDAC6, A MEMBER OF THE HDAC FAMILY, WHOSE MAJOR SUBSTRATE IS ALPHA-TUBULIN, IS BEING INCREASINGLY IMPLICATED IN THE PATHOGENESIS OF INFLAMMATORY DISORDERS. THE PRESENT STUDY WAS CARRIED OUT TO STUDY THE POTENTIAL ANTI-INFLAMMATORY AND ANTI-RHEUMATIC EFFECTS OF HDAC6 SELECTIVE INHIBITOR TUBASTATIN. TUBASTATIN, A POTENT HUMAN HDAC6 INHIBITOR WITH AN IC50 OF 11 NM SHOWED SIGNIFICANT INHIBITION OF TNF-ALPHA AND IL-6 IN LPS STIMULATED HUMAN THP-1 MACROPHAGES WITH AN IC50 OF 272 NM AND 712 NM RESPECTIVELY. ADDITIONALLY, TUBASTATIN INHIBITED NITRIC OXIDE (NO) SECRETION IN MURINE RAW 264.7 MACROPHAGES DOSE DEPENDENTLY WITH AN IC50 OF 4.2 MUM AND INDUCED ALPHA-TUBULIN HYPERACETYLATION CORRESPONDING TO HDAC6 INHIBITION IN THP-1 CELLS WITHOUT AFFECTING THE CELL VIABILITY. TUBASTATIN SHOWED SIGNIFICANT INHIBITION OF PAW VOLUME AT 30 MG/KG I.P. IN A FREUND'S COMPLETE ADJUVANT (FCA) INDUCED ANIMAL MODEL OF INFLAMMATION. THE DISEASE MODIFYING ACTIVITY OF TUBASTATIN WAS ALSO EVIDENT IN COLLAGEN INDUCED ARTHRITIS DBA1 MOUSE MODEL AT 30 MG/KG I.P. THE SIGNIFICANT ATTENUATION OF CLINICAL SCORES (~70%) BY TUBASTATIN WAS CONFIRMED HISTOPATHOLOGICALLY AND WAS FOUND COMPARABLE TO DEXAMETHASONE (~90% INHIBITION OF CLINICAL SCORES). TUBASTATIN SHOWED SIGNIFICANT INHIBITION OF IL-6 IN PAW TISSUES OF ARTHRITIC MICE. THE PRESENT WORK HAS DEMONSTRATED ANTI-INFLAMMATORY AND ANTIRHEUMATIC EFFECTS OF A SELECTIVE HDAC6 INHIBITOR TUBASTATIN. 2013 5 1471 32 DISTINCT GLOBAL DNA METHYLATION STATUS IN B-CELL LYMPHOMAS: IMMUNOHISTOCHEMICAL STUDY OF 5-METHYLCYTOSINE AND 5-HYDROXYMETHYLCYTOSINE. LYMPHOMAS ARE MALIGNANT NEOPLASMS COMPOSED OF LYMPHOID CELLS AT VARIOUS DEVELOPMENTAL STAGES AND LINEAGES. RECENT ADVANCES IN COMPREHENSIVE GENOMIC ANALYSES IN ACUTE MYELOID LEUKEMIA HAVE REVEALED PREVALENT MUTATIONS IN REGULATORS OF EPIGENETIC PHENOMENA INCLUDING GLOBAL DNA METHYLATION STATUS. THE EXAMPLES INCLUDE MUTATIONS IN ISOCITRATE DEHYDROGENASE 1 (IDH1), IDH2, AND TEN-ELEVEN TRANSLOCATION 2. THESE MUTATIONS ARE PROPOSED TO INHIBIT CONVERSION OF 5-METHYLCYTOSINE (5 MC) TO 5-HYDROXYMETHYLCYTOSINE (5 HMC), LEADING TO GLOBAL ACCUMULATION OF 5 MC. THESE CHANGES IN GLOBAL DNA METHYLATION STATUS CAN BE VISUALIZED IMMUNOHISTOCHEMICALLY USING SPECIFIC ANTIBODIES AGAINST 5 MC AND 5 HMC. WE EXAMINED THE GLOBAL DNA METHYLATION STATUS OF B-CELL LYMPHOMAS AND THAT OF THEIR NORMAL COUNTERPARTS BY IMMUNOHISTOCHEMISTRY FOR 5 MC AND 5 HMC. NON-TUMOR LYMPHOID CELLS INSIDE GERMINAL CENTERS (GC) IN REACTIVE LYMPHOID HYPERPLASIA (RLH) WERE STAINED POSITIVE FOR 5 MC, BUT THEY WERE NEGATIVE FOR 5 HMC. SIMILARLY, FOLLICULAR LYMPHOMAS, WHOSE POSTULATED NORMAL COUNTERPARTS ARE CENTROCYTES IN GCS, WERE 5 MC-POSITIVE BUT 5 HMC-NEGATIVE BY IMMUNOHISTOCHEMISTRY. THIS IMMUNOSTAINING PATTERN WAS ALSO OBSERVED IN BURKITT LYMPHOMA. IN CONTRAST, NON-TUMOR LYMPHOID CELLS IN MANTLE ZONES WERE STAINED POSITIVE FOR 5 MC AS WELL AS FOR 5 HMC. LIKEWISE, MOST MANTLE CELL LYMPHOMAS, WHOSE POSTULATED NORMAL COUNTERPARTS ARE MANTLE ZONE B CELLS IN RLH, WERE STAINED POSITIVE FOR 5 MC AS WELL AS FOR 5 HMC. THIS IMMUNOSTAINING PATTERN WAS ALSO OBSERVED IN CHRONIC LYMPHOCYTIC LEUKEMIA/SMALL LYMPHOCYTIC LYMPHOMA. THESE RESULTS SUGGEST THAT, IN TERMS OF 5 MC/5 HMC IMMUNOHISTOCHEMISTRY, B-CELL LYMPHOMAS WITH DIFFERENT HISTOLOGICAL SUBTYPES ARE ASSOCIATED WITH DISTINCT GLOBAL DNA METHYLATION STATUSES THAT RESEMBLE THOSE OF THEIR POSTULATED NORMAL COUNTERPARTS. 2014 6 1572 24 DNA METHYLATION PATTERNS IN JUVENILE SYSTEMIC SCLEROSIS AND LOCALIZED SCLERODERMA. SCLERODERMA REFERS TO A GROUP OF CHRONIC FIBROTIC IMMUNE-MEDIATED DISEASES OF UNKNOWN ETIOLOGY. CHARACTERIZING EPIGENETIC CHANGES IN CHILDHOOD-ONSET SCLERODERMA, SYSTEMIC SCLEROSIS OR LOCALIZED SCLERODERMA, HAS NOT BEEN PREVIOUSLY PERFORMED. THE AIM OF THIS STUDY WAS TO ASSESS DNA METHYLATION DIFFERENCES AND SIMILARITIES BETWEEN JUVENILE SYSTEMIC SCLEROSIS (JSSC) AND JUVENILE LOCALIZED SCLERODERMA (JLS) COMPARED TO MATCHED HEALTHY CONTROLS. GENOME-WIDE DNA METHYLATION CHANGES IN PERIPHERAL BLOOD MONONUCLEAR CELL SAMPLES WERE ASSESSED USING THE METHYLATIONEPIC ARRAY FOLLOWED BY BIOINFORMATIC ANALYSIS AND LIMITED FUNCTIONAL ASSESSMENT. WE IDENTIFIED A TOTAL OF 105 AND 144 DIFFERENTIALLY METHYLATED SITES COMPARED TO HEALTHY CONTROLS IN JSSC AND JLS, RESPECTIVELY. THE MAJORITY OF DIFFERENTIALLY METHYLATED SITES AND GENES REPRESENTED WERE UNIQUE TO EITHER JSSC OR JLS SUGGESTING A DIFFERENT UNDERLYING EPIGENETIC PATTERN IN BOTH DISEASES. AMONG SHARED DIFFERENTIALLY METHYLATED GENES, METHYLATION LEVELS IN A CPG SITE IN FGFR2 CAN DISTINGUISH BETWEEN LS AND HEALTHY PBMCS WITH A HIGH ACCURACY. CANONICAL PATHWAY ANALYSIS REVEALED THAT INFLAMMATORY PATHWAYS WERE ENRICHED IN GENES DIFFERENTIALLY METHYLATED IN JSSC, INCLUDING STAT3, NF-KAPPAB, AND IL-15 PATHWAYS. IN CONTRAST, THE HIPPO SIGNALING PATHWAY WAS ENRICHED IN JLS. OUR DATA ALSO SUGGEST A POTENTIAL ROLE FOR NOTCH3 IN BOTH JSSC AND JLS, AND REVEALED A NUMBER OF TRANSCRIPTION FACTORS UNIQUE TO EACH OF THE TWO DISEASES. IN SUMMARY, OUR DATA REVEALED IMPORTANT INSIGHTS INTO JSSC AND JLS AND SUGGEST A POTENTIALLY NOVEL EPIGENETIC DIAGNOSTIC BIOMARKER FOR LS. 2021 7 2311 33 EPIGENETIC REGULATION OF CYTOSOLIC PHOSPHOLIPASE A2 IN SH-SY5Y HUMAN NEUROBLASTOMA CELLS. GROUP IVA CYTOSOLIC PHOSPHOLIPASE A2 (CPLA2 OR PLA2G4A) IS A KEY ENZYME THAT CONTRIBUTES TO INFLAMMATION VIA THE GENERATION OF ARACHIDONIC ACID AND EICOSANOIDS. WHILE MUCH IS KNOWN ABOUT REGULATION OF CPLA2 BY POSTTRANSLATIONAL MODIFICATION SUCH AS PHOSPHORYLATION, LITTLE IS KNOWN ABOUT ITS EPIGENETIC REGULATION. IN THIS STUDY, TREATMENT WITH HISTONE DEACETYLASE (HDAC) INHIBITORS, TRICHOSTATIN A (TSA), VALPROIC ACID, TUBACIN AND THE CLASS I HDAC INHIBITOR, MS-275, WERE FOUND TO INCREASE CPLA2ALPHA MESSENGER RNA (MRNA) EXPRESSION IN SH-SY5Y HUMAN NEUROBLASTOMA CELLS. CO-TREATMENT OF THE HISTONE ACETYLTRANSFERASE (HAT) INHIBITOR, ANACARDIC ACID, MODULATED UPREGULATION OF CPLA2ALPHA INDUCED BY TSA. SPECIFIC INVOLVEMENT OF CLASS I HDACS AND HAT IN CPLA2ALPHA REGULATION WAS FURTHER SHOWN, AND A TIP60-SPECIFIC HAT INHIBITOR, NU9056, MODULATED THE UPREGULATION OF CPLA2ALPHA INDUCED BY MS-275. IN ADDITION, CO-TREATMENT OF WITH HISTONE METHYLTRANSFERASE (HMT) INHIBITOR, 5'-DEOXY-5'-METHYLTHIOADENOSINE (MTA) SUPPRESSED TSA-INDUCED CPLA2ALPHA UPREGULATION. THE ABOVE CHANGES IN CPLA2 MRNA EXPRESSION WERE REFLECTED AT THE PROTEIN LEVEL BY WESTERN BLOTS AND IMMUNOCYTOCHEMISTRY. CHROMATIN IMMUNOPRECIPITATION (CHIP) SHOWED TSA INCREASED BINDING OF TRIMETHYLATED H3K4 TO THE PROXIMAL PROMOTER REGION OF THE CPLA2ALPHA GENE. CELL INJURY AFTER TSA TREATMENT AS INDICATED BY LACTATE DEHYDROGENASE (LDH) RELEASE WAS MODULATED BY ANACARDIC ACID, AND A ROLE OF CPLA2 IN MEDIATING TSA-INDUCED INJURY SHOWN, AFTER CO-INCUBATION WITH THE CPLA2 SELECTIVE INHIBITOR, ARACHIDONOYL TRIFLUOROMETHYL KETONE (AACOCF3). TOGETHER, RESULTS INDICATE EPIGENETIC REGULATION OF CPLA2 AND THE POTENTIAL OF SUCH REGULATION FOR TREATMENT OF CHRONIC INFLAMMATION. 2016 8 4054 20 MAPK IS A MUTUAL PATHWAY TARGETED BY ANXIETY-RELATED MIRNAS, AND E2F5 IS A PUTATIVE TARGET FOR ANXIOLYTIC MIRNAS. ANXIETY-RELATED DISORDERS (ARDS) ARE CHRONIC NEUROPSYCHOLOGICAL DISEASES AND THE SIXTH LEADING CAUSE OF DISABILITY IN THE WORLD. AS DYSREGULATION OF MICRORNAS (MIRS) ARE OBSERVED IN THE PATHOLOGICAL COURSE OF NEUROPSYCHIATRIC DISORDERS, THE PRESENT STUDY AIMED TO INTRODUCE MIRS THAT UNDERLIE ANXIETY PROCESSING IN THE BRAIN. FIRST, WE COLLECTED THE EXPERIMENTALLY CONFIRMED ANXIETY-RELATED MIRNAS (ARMIRS), PREDICTED THEIR TARGET TRANSCRIPTS, AND INTRODUCED CRITICAL CELLULAR PATHWAYS WITH KEY COMMUNE HUB GENES. AS A RESULT, WE HAVE FOUND NINE ANXIOLYTIC AND TEN ANXIOGENIC ARMIRS. THE ANXIOLYTIC MIRS FREQUENTLY TARGET THE MRNA OF ACYL-COA SYNTHETASE LONG-CHAIN FAMILY MEMBER 4 (ACSL4), AFF4-AF4/FMR2 FAMILY MEMBER 4 (AFF4), AND KRUPPEL LIKE TRANSCRIPTION FACTOR 4 (KLF4) GENES, WHERE MIR-34B-5P AND MIR-34C-5P INTERACT WITH ALL OF THEM. MOREOVER, THE ANXIOGENIC MIRS FREQUENTLY TARGET THE MRNA OF NINE GENES; AMONG THEM, ONLY TWO MIR (MIR-142-5P AND MIR-218-5P) HAVE NO INTERACTION WITH THE MRNA OF TRINUCLEOTIDE REPEAT-CONTAINING ADAPTOR 6B (TNRC6B), AND MIR-124-3P INTERACTS WITH ALL OF THEM WHERE MAPK IS THE MAIN SIGNALING PATHWAY AFFECTED BY BOTH ANXIOLYTIC AND ANXIOGENIC MIR. IN ADDITION, THE ANXIOLYTIC MIR COMMONLY TARGET E2F TRANSCRIPTION FACTOR 5 (E2F5) IN THE TGF-BETA SIGNALING PATHWAY, AND THE ANXIOGENIC MIR COMMONLY TARGET ATAXIN 1 (ATXN1), WASP-LIKE ACTIN NUCLEATION PROMOTING FACTOR (WASL), AND SOLUTE CARRIER FAMILY 17 MEMBER 6 (SLC17A6) GENES IN THE NOTCH SIGNALING, ADHERENCE JUNCTION, AND SYNAPTIC VESICLE CYCLE PATHWAYS, RESPECTIVELY. TAKEN TOGETHER, WE CONCLUDE THAT THE MOST IMPORTANT ANXIOLYTIC (MIR-34C, LET-7D, AND MIR-17) AND ANXIOGENIC (MIR-19B, MIR-92A, AND 218) MIR, AS HUB EPIGENETIC MODULATORS, POTENTIALLY INFLUENCE THE PATHOPHYSIOLOGY OF ANXIETY, PRIMARILY VIA INTERACTION WITH THE MAPK SIGNALING PATHWAY. MOREOVER, THE ROLE OF E2F5 AS A NOVEL PUTATIVE TARGET FOR ANXIOLYTIC MIRNAS IN ARDS DISORDERS DESERVES FURTHER EXPLORATION. 2023 9 2749 29 EXPRESSION AND POLYMORPHISM OF MICRO-RNA ACCORDING TO BODY MASS INDEX AND BREAST CANCER PRESENTATION IN TUNISIAN PATIENTS. MICRO-RNAS (MIRS) CONSTITUTE A CLASS OF SMALL NONCODING RNAS IMPLICATED IN THE REGULATION OF GENE EXPRESSION BY BINDING TO TARGET MRNAS. A MIR CAN TARGET SEVERAL MRNAS, BEING INVOLVED IN DIFFERENT BIOLOGIC PROCESSES AND PATHOLOGIES. THIS PLEIOTROPIC FUNCTION MIGHT EXPLAIN THE LINK BETWEEN DISEASES CO-OCCURRENCE. EPIGENETIC ORIGIN OF THE LINK BETWEEN OBESITY AND BREAST CANCER (BC) IS INVESTIGATED IN A COHORT OF TUNISIAN PATIENTS, FOCUSING ON POLYMORPHISM AT GERMLINE LEVEL (MIR-146A) AND ON EXPRESSION IN MAMMARY TUMORS (MIR-21, MIR-146A, AND MIR-34A), ACCORDING TO BODY MASS INDEX (BMI) AND CLINICO-PATHOLOGIC FEATURES. THE MEASURE OF MIR EXPRESSION IN 60 MAMMARY TUMORS WAS REALIZED USING QUANTITATIVE RT-PCR. STUDY OF RS 2910164 IN MIR-146A WAS PERFORMED BY PCR AND DIRECT SEQUENCING USING BLOOD DNA OF 83 AFFECTED WOMEN AND 50 UNRELATED SUBJECTS FROM GREAT TUNIS. MIR-21, MIR-146A, AND MIR-34A HAVE BEEN QUANTIFIED IN BREAST TUMOR ACCORDING TO BMI. MIR-21 IS SIGNIFICANTLY MORE EXPRESSED IN TUMORS OF OBESE WOMEN COMPARATIVELY TO NONOBESE PATIENTS. ON THE CONTRARY, MIR-34A IS DECREASED IN TUMORS OF OBESE WOMEN. MOREOVER, IN OBESE BC PATIENTS, A SIGNIFICANT INCREASE IN BOTH MIR-21 AND MIR-146A EXPRESSION IS REVEALED IN CASES WITH LYMPH NODE METASTASIS. THE POLYMORPHISM AT RS 2910164 (MIR-146A) LOCUS WAS NOT SHOWN AS A RISK FACTOR FOR BC. HOWEVER THE MUTANT CC GENOTYPE WAS REVEALED TO BE ASSOCIATED WITH A RISK FOR BAD OUTCOME OF THE DISEASE. CHRONIC INFLAMMATION IN OBESE WOMEN WOULD BE LINKED TO AGGRESSIVE BREAST TUMORS VIA INDUCTION OF ONCOMIRS OVEREXPRESSION AND DECREASE OF TUMOR SUPPRESSOR MIRS. 2019 10 4350 31 MIR-181A-5P IS A POTENTIAL CANDIDATE EPIGENETIC BIOMARKER IN MULTIPLE SCLEROSIS. MULTIPLE SCLEROSIS (MS) IS A CHRONIC INFLAMMATORY DISEASE OF THE CENTRAL NERVOUS SYSTEM (CNS) CHARACTERIZED BY DEMYELINATION AND AXONAL DEGENERATION. ABNORMAL EXPRESSION OF MICRORNAS (MIRNAS) PLAYS AN IMPORTANT ROLE IN MS PATHOLOGY. IN THIS COHORT STUDY, DIFFERENTIAL EXPRESSION OF THE FOUR MIRNAS (HSA-MIR-155-5P, HSA-MIR-9-5P, HSA-MIR-181A-5P, AND HSA-MIR-125B-5P) WAS INVESTIGATED IN 69 INDIVIDUALS, INCLUDING 39 MS PATIENTS (RELAPSING-REMITTING MS (RRMS), N = 27; SECONDARY PROGRESSIVE MS (SPMS), N = 12) AND 30 HEALTHY CONTROLS. IN SILICO ANALYSES REVEALED POSSIBLE GENES AND PATHWAYS SPECIFIC TO MIRNAS. PERIPHERAL BLOOD MIRNA EXPRESSIONS WERE DETECTED BY QUANTITATIVE REAL-TIME PCR (QPCR). HSA-MIR-181A-5P WAS DOWNREGULATED AND ASSOCIATED WITH INCREASED MS RISK (P = 0.012). THE OTHER THREE MIRNAS WERE UPREGULATED AND NOT ASSOCIATED WITH MS (P < 0.05). THE AREA UNDER THE CURVE (AUC) IS 0.779. IN SILICO ANALYSES SHOWED THAT HSA-MIR-181A-5P MAY PARTICIPATE IN MS PATHOLOGY BY TARGETING MAP2K1, CREB1, ATXN1, AND ATXN3 GENES IN INFLAMMATION AND NEURODEGENERATION PATHWAYS. THE CIRCULATORY HSA-MIR-181A-5P CAN REGULATE TARGET GENES, REVERSING THE MECHANISMS INVOLVED IN MS PATHOLOGIES SUCH AS PROTEIN UPTAKE AND PROCESSING, CELL PROLIFERATION AND SURVIVAL, INFLAMMATION, AND NEURODEGENERATION. THUS, THIS MIRNA COULD BE USED AS AN EPIGENOMIC-GUIDED DIAGNOSTIC TOOL AND FOR THERAPEUTIC PURPOSE. 2022 11 5479 30 RESVERATROL ATTENUATES CIGARETTE SMOKE EXTRACT INDUCED CELLULAR SENESCENCE IN HUMAN AIRWAY EPITHELIAL CELLS BY REGULATING THE MIR-34A/SIRT1/NF-KAPPAB PATHWAY. CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) IS CHARACTERIZED BY ACCELERATED LUNG AGING. SMOKING IS THE CRITICAL RISK FACTOR FOR COPD. CELLULAR SENESCENCE OF AIRWAY EPITHELIAL CELLS IS THE CYTOLOGICAL BASIS OF ACCELERATED LUNG AGING IN COPD, AND THE REGULATION OF MICRORNAS (MIRNAS) IS THE CENTRAL EPIGENETIC MECHANISM OF CELLULAR SENESCENCE. RESVERATROL (RES) IS A POLYPHENOL WITH ANTI-AGING PROPERTIES. THIS STUDY INVESTIGATED WHETHER RES ATTENUATES CIGARETTE SMOKE EXTRACT (CSE)-INDUCED CELLULAR SENESCENCE IN HUMAN AIRWAY EPITHELIAL CELLS (BEAS-2B) THROUGH THE MIR-34A/SIRT1/NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) PATHWAY. BEAS-2B CELLS WERE TREATED WITH RES, CSE AND TRANSFECTED WITH MIR-34A-5P MIMICS. CELLULAR SENESCENCE WAS EVALUATED BY SENESCENCE -RELATED BETA-GALACTOSIDASE (SA-BETA-GAL) STAINING AND EXPRESSION OF SENESCENCE-RELATED GENES (P16, P21, AND P53). THE EXPRESSIONS OF MIR-34A-5P, SIRT1, AND NF-KAPPAB P65 WERE EXAMINED USING QUANTITATIVE REAL TIME POLYMERASE CHAIN REACTION AND WESTERN BLOTTING. THE SENESCENCE-ASSOCIATED SECRETORY PHENOTYPE (SASP) CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) WERE ASSESSED BY ENZYME-LINKED IMMUNOSORBENT ASSAY. THE BINDING BETWEEN MIR-34A-5P AND SIRT1 WAS CONFIRMED BY DUAL-LUCIFERASE REPORTER ASSAY. THE RESULTS SHOWED THAT CSE DOSE-DEPENDENTLY DECREASED CELL VIABILITY AND ELEVATED CELLULAR SENESCENCE, CHARACTERIZED BY INCREASED SA-BETA-GAL STAINING AND SENESCENCE-RELATED GENE EXPRESSIONS (P16, P21, AND P53). FURTHER, CSE DOSE-DEPENDENTLY INCREASED THE EXPRESSION OF MIR-34A-5P AND SASP CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) IN BEAS-2B CELLS. PRETREATMENT WITH RES INHIBITED CSE-INDUCED CELLULAR SENESCENCE AND SECRETION OF SASP CYTOKINES (IL-1BETA, IL-6, IL-8, TNF-ALPHA) IN A DOSE-DEPENDENT MANNER. MOREOVER, RES REVERSED THE CSE-INDUCED DOWN-REGULATION OF SIRT1 AND UP-REGULATION OF MIR-34A-5P AND NF-KAPPAB P65. SIRT1 IS A TARGET OF MIR-34A-5P. OVEREXPRESSION OF MIR-34A-5P VIA TRANSFECTION WITH MIR-34A-5P MIMIC IN BEAS-2B CELLS ATTENUATED THE INHIBITORY EFFECT OF RES ON CELLULAR SENESCENCE, ACCOMPANIED BY REVERSING THE EXPRESSION OF SIRT1 AND NF-KAPPAB P65. IN CONCLUSION, RES ATTENUATED CSE-INDUCED CELLULAR SENESCENCE IN BEAS-2B CELLS BY REGULATING THE MIR-34A/SIRT1/NF-KAPPAB PATHWAY, WHICH MAY PROVIDE A NEW APPROACH FOR COPD TREATMENT. 2022 12 5891 27 SYSTEMS BIOLOGY IN CHRONIC HEART FAILURE-IDENTIFICATION OF POTENTIAL MIRNA REGULATORS. HEART FAILURE (HF) IS A COMPLEX DISEASE ENTITY WITH HIGH CLINICAL IMPACT, POORLY UNDERSTOOD PATHOPHYSIOLOGY AND SCANTLY KNOWN MIRNA-MEDIATED EPIGENETIC REGULATION. WE HAVE ANALYSED MIRNA PATTERNS IN PATIENTS WITH CHRONIC HF (CHF) AND A SEX- AND AGE-MATCHED REFERENCE GROUP AND PURSUED AN IN SILICO SYSTEM BIOLOGY ANALYSIS TO DISCERN PATHWAYS INVOLVED IN CHF PATHOPHYSIOLOGY. TWENTY-EIGHT MIRNAS WERE IDENTIFIED IN CHF THAT WERE UP-REGULATED IN THE REFERENCE GROUP, AND EIGHT OF THEM WERE VALIDATED BY RT-QPCR. IN SILICO ANALYSIS OF PREDICTED TARGETS BY STRING PROTEIN-PROTEIN INTERACTION NETWORKS REVEALED EIGHT CLUSTER NETWORKS (INVOLVING SEVEN OF THE IDENTIFIED MIRNAS) ENRICHED IN PATHWAYS RELATED TO CELL CYCLE, RAS, CHEMOKINE, PI3K-AKT AND TGF-BETA SIGNALING. BY ROC CURVE ANALYSIS, COMBINED PROBABILITIES OF THESE SEVEN MIRNAS (LET-7A-5P, MIR-107, MIR-125A-5P, MIR-139-5P, MIR-150-5P, MIR-30B-5P AND MIR-342-3P; CLUSTERS 1-4 [C:1-4]), DISCRIMINATED BETWEEN HF WITH PRESERVED EJECTION FRACTION (HFPEF) AND HF WITH REDUCED EJECTION FRACTION (HFREF), AND ISCHAEMIC AND NON-ISCHAEMIC AETIOLOGY. A COMBINATION OF MIR-107, MIR-139-5P AND MIR-150-5P, INVOLVED IN CLUSTERS 5 AND 7 (C:5+7), DISCRIMINATED HFPEF FROM HFREF. PATHWAY ENRICHMENT ANALYSIS OF MIRNAS PRESENT IN C:1-4 (LET-7A-5P, MIR-125A-5P, MIR-30B-5P AND MIR-342-3P) REVEALED PATHWAYS RELATED TO HF PATHOGENESIS. IN CONCLUSION, WE HAVE IDENTIFIED A DIFFERENTIAL SIGNATURE OF DOWN-REGULATED MIRNAS IN THE PLASMA OF HF PATIENTS AND PROPOSE NOVEL CELLULAR MECHANISMS INVOLVED IN CHF PATHOGENESIS. 2022 13 908 26 CHRONIC EXPOSURE TO ENVIRONMENTALLY RELEVANT CONCENTRATION OF FLUORIDE IMPAIRS OSTEOBLAST'S COLLAGEN SYNTHESIS AND MATRIX MINERALIZATION: INVOLVEMENT OF EPIGENETIC REGULATION IN SKELETAL FLUOROSIS. GLOBALLY, 200 MILLION PEOPLE ARE SUFFERING FROM TOXIC MANIFESTATIONS OF FLUORIDE(F), DENTAL AND SKELETAL FLUOROSIS; UNFORTUNATELY, THERE IS NO TREATMENT. TO UNRAVEL THE PATHOGENESIS OF SKELETAL FLUOROSIS, WE ESTABLISHED FLUOROSIS MICE BY TREATING ENVIRONMENTALLY RELEVANT CONCENTRATION OF F (15 PPM NAF) THROUGH DRINKING WATER FOR 4 MONTHS. AS IN SKELETAL FLUOROSIS, LOCOMOTOR DISABILITY, CRIPPLING DEFORMITIES OCCUR AND THUS, OUR HYPOTHESIS WAS F MIGHT ADVERSELY AFFECTS COLLAGEN WHICH GIVES THE BONE TENSILE STRENGTH. THIS WORK INEVITABLY HAD TO BE CARRIED OUT ON OSTEOBLAST CELLS, RESPONSIBLE FOR SYNTHESIS, DEPOSITION, AND MINERALIZATION OF BONE MATRIX. ISOLATED OSTEOBLAST CELLS WERE CONFIRMED BY ALP ACTIVITY AND MINERALIZED NODULES FORMATION. EXPRESSION OF COLLAGEN COL1A1, COL1A2, COL1A1 WAS SIGNIFICANTLY REDUCED IN TREATED MICE. FURTHER, A STUDY REVEALED THE INVOLVEMENT OF EPIGENETIC REGULATION BY PROMOTER HYPERMETHYLATION OF COL1A1; EXPRESSIONAL ALTERATIONS OF TRANSCRIPTION FACTORS, CALCIUM CHANNELS AND OTHER GENES E.G., CBFA-1, TGF-BETA1, BMP1, SP1, SP7, NF-(K)B P65, BMP-2, BGLAP, GPRC6A AND CAV(1.2) ARE ASSOCIATED WITH IMPAIRMENT OF COLLAGEN SYNTHESIS, DEPOSITION AND DECREASED MINERALIZATION THUS, ENFEEBLING BONE HEALTH. THIS STUDY INDICATES THE POSSIBLE ASSOCIATION OF EPIGENETIC REGULATION IN SKELETAL FLUOROSIS. HOWEVER, NO ASSOCIATION WAS FOUND BETWEEN POLYMORPHISMS IN THE COL1A1 (RSAI, HINDIII) AND COL1A2 (RSAI, HINDIII) GENES WITH FLUOROSIS IN MICE. 2023 14 4301 23 MICRORNA-21-ENRICHED EXOSOMES AS EPIGENETIC REGULATORS IN MELANOMAGENESIS AND MELANOMA PROGRESSION: THE IMPACT OF WESTERN LIFESTYLE FACTORS. DNA MUTATION-INDUCED ACTIVATION OF RAS-BRAF-MEK-ERK SIGNALING ASSOCIATED WITH INTERMITTENT OR CHRONIC ULTRAVIOLET (UV) IRRADIATION CANNOT EXCLUSIVELY EXPLAIN THE EXCESSIVE INCREASE OF MALIGNANT MELANOMA (MM) INCIDENCE SINCE THE 1950S. MALIGNANT CONVERSION OF A MELANOCYTE TO AN MM CELL AND METASTATIC MM IS ASSOCIATED WITH A STEADY INCREASE IN MICRORNA-21 (MIR-21). AT THE EPIGENETIC LEVEL, MIR-21 INHIBITS KEY TUMOR SUPPRESSORS OF THE RAS-BRAF SIGNALING PATHWAY ENHANCING PROLIFERATION AND MM PROGRESSION. INCREASED MM CELL LEVELS OF MIR-21 EITHER RESULT FROM ENDOGENOUS UPREGULATION OF MELANOCYTIC MIR-21 EXPRESSION OR BY UPTAKE OF MIR-21-ENRICHED EXOGENOUS EXOSOMES. BASED ON EPIDEMIOLOGICAL DATA AND TRANSLATIONAL EVIDENCE, THIS REVIEW PROVIDES DEEPER INSIGHTS INTO ENVIRONMENTALLY AND METABOLICALLY INDUCED EXOSOMAL MIR-21 TRAFFICKING BEYOND UV-IRRADIATION IN MELANOMAGENESIS AND MM PROGRESSION. SOURCES OF MIR-21-ENRICHED EXOSOMES INCLUDE UV-IRRADIATED KERATINOCYTES, ADIPOCYTE-DERIVED EXOSOMES IN OBESITY, AIRWAY EPITHELIUM-DERIVED EXOSOMES GENERATED BY SMOKING AND POLLUTION, DIET-RELATED EXOSOMES AND INFLAMMATION-INDUCED EXOSOMES, WHICH MAY SYNERGISTICALLY INCREASE THE EXOSOMAL MIR-21 BURDEN OF THE MELANOCYTE, THE TRANSFORMED MM CELL AND ITS TUMOR ENVIRONMENT. SEVERAL THERAPEUTIC AGENTS THAT SUPPRESS MM CELL GROWTH AND PROLIFERATION ATTENUATE MIR-21 EXPRESSION. THESE INCLUDE MIR-21 ANTAGONISTS, METFORMIN, KINASE INHIBITORS, BETA-BLOCKERS, VITAMIN D, AND PLANT-DERIVED BIOACTIVE COMPOUNDS, WHICH MAY REPRESENT NEW OPTIONS FOR THE PREVENTION AND TREATMENT OF MM. 2020 15 4304 27 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 5426 30 REGULATION OF SIRTUIN EXPRESSION IN AUTOIMMUNE NEUROINFLAMMATION: INDUCTION OF SIRT1 IN OLIGODENDROCYTE PROGENITOR CELLS. IN MULTIPLE SCLEROSIS (MS) REGENERATION OF OLIGODENDROCYTES FOLLOWING INFLAMMATORY DEMYELINATION IS LIMITED BY THE COMPROMISED ABILITY OF PROGENITORS TO REPOPULATE LESIONED AREAS AND TRANSITION TO FUNCTIONALLY COMPETENT OLIGODENDROCYTES. REGARDING UNDERLYING MECHANISMS, THE INVOLVEMENT OF EPIGENETIC PROCESSES HAS BEEN SUGGESTED, E.G. THE CONTRIBUTION OF HISTONE DEACETYLASES (HDAC) KNOWN TO REGULATE OLIGODENDROCYTE PROGENITOR CELL (OPC) DIFFERENTIATION. HOWEVER, THEIR PRECISE EXPRESSION PATTERNS, PARTICULAR OF REDOX-SENSITIVE NAD(+) HDACS, REMAINS LARGELY UNKNOWN. IN THIS STUDY, WE DETERMINED THE EXPRESSION AND ACTIVITY OF SIRTUINS, MEMBERS OF THE HDAC CLASS III FAMILY WITH A SPECIFIC FOCUS ON SIRT1, PREVIOUSLY ASSOCIATED WITH NEURODEGENERATIVE, INFLAMMATORY AND DEMYELINATING DISORDERS OF THE CENTRAL NERVOUS SYSTEM (CNS). BY INVESTIGATING MOUSE EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE), A MODEL FOR MS, WE FOUND THAT TRANSCRIPTION OF SIRT1, SIRT2 AND SIRT6 WAS SIGNIFICANTLY INCREASED IN THE CNS DURING CHRONIC DISEASE STAGES. WE CONFIRMED THIS FINDING FOR SIRT1 PROTEIN EXPRESSION AND WERE ABLE TO LOCALIZE UPREGULATED SIRT1 IN NUCLEI OF NG2(+) OR PDGFRALPHA(+) OPCS IN DEMYELINATED BRAIN LESIONS. IN CULTURED MOUSE A2B5(+) OPCS BLOCKADE OF SIRT1 ACTIVITY BY THE SMALL MOLECULE COMPOUND EX527 ENHANCED MITOTIC ACTIVITY BUT DID NOT AFFECT THE CAPACITY TO DIFFERENTIATE. A SIMILAR PATTERN WAS DETECTABLE IN OPCS DERIVED FROM SIRT1-DEFICIENT ANIMALS. TAKEN TOGETHER, OUR DATA SUGGEST THAT SIRT1 INHIBITION MAY HELP TO EXPAND THE ENDOGENOUS POOL OF OPCS WITHOUT AFFECTING THEIR DIFFERENTIATION. 2019 17 6217 30 THE JAK INHIBITOR TOFACITINIB INHIBITS STRUCTURAL DAMAGE IN OSTEOARTHRITIS BY MODULATING JAK1/TNF-ALPHA/IL-6 SIGNALING THROUGH MIR-149-5P. BACKGROUND: OSTEOARTHRITIS (OA), A COMMON ARTICULAR BONE DEGENERATIVE DISEASE, IS EXACERBATED BY PROINFLAMMATORY CYTOKINE SIGNALING. MOUNTING EVIDENCE SUGGESTS THAT EPIGENETIC MODIFIERS, NAMELY MICRORNAS (MIRS), ARE DYSREGULATED IN ARTICULAR CHONDROCYTES (ACS) DURING OA. METHODS: AN INITIAL DATABASE SEARCH LED TO THE IDENTIFICATION OF MIR-149-5P, WHICH WAS DOWNREGULATED IN CLINICAL OA SAMPLES AND CONTRIBUTED TO CHRONIC INFLAMMATION, BY INCREASING TNF-ALPHA/IL-6 SIGNALING WITHIN THE SYNOVIUM, AND OA PROGRESSION. RESULTS: WE OVEREXPRESSED MIR-149-5P IN THE HUMAN CHONDROCYTE CELL LINES C20A4 AND C28/I2 TO EXAMINE ITS ROLE IN CHONDROCYTE HYPERTROPHY AND OSTEOCLASTOGENESIS AND FOUND A SIGNIFICANT DECREASE IN IL-6 EXPRESSION, AN INCREASE IN SOX9 EXPRESSION, AND A REDUCTION IN CHONDROCYTE HYPERTROPHY. WE EVALUATED THE THERAPEUTIC EFFECTS OF TOFACITINIB (JAK INHIBITOR) BY SUPPRESSING INFLAMMATION AND RESTORING MIR-149-5P EXPRESSION. TOFACITINIB-TREATED C20A4 AND C28/I2 CELLS HAD A SIGNIFICANTLY LOWER EXPRESSION OF JAK/IL-6/TNF-ALPHA AND AN INCREASED LEVEL OF MIR-149-5P. NOTABLY, TOFACITINIB TREATMENT REDUCED AC HYPERTROPHY AND SECRETION OF RANKL AND IL-6. FINALLY, AN OA MOUSE MODEL WAS USED TO EVALUATE THE THERAPEUTIC POTENTIAL OF TOFACITINIB. INTRA-ARTICULAR INJECTION OF TOFACITINIB SIGNIFICANTLY LOWERED ARTHRITIS SCORES AND BONE DEGRADATION IN TREATED MICE COMPARED WITH THEIR CONTROL COUNTERPARTS. CONCLUSION: WE SHOW FOR THE FIRST TIME THAT TOFACITINIB SUPPRESSES THE EXPRESSION LEVEL OF JAK1/TNF-ALPHA/IL-6 BY UPREGULATING MIR-149-5P LEVEL. OUR FINDINGS REVEALED THE FUNCTIONAL ASSOCIATION BETWEEN PROINFLAMMATORY JAK1/TNF-ALPHA/IL-6 SIGNALING AND ACS DEVELOPMENT AND HIGHLIGHT THE THERAPEUTIC POTENTIAL OF TOFACITINIB IN OA. 2021 18 5120 26 POSSIBLE EPIGENETIC REGULATORY EFFECT OF DYSREGULATED CIRCULAR RNAS IN EPILEPSY. CIRCULAR RNAS (CIRCRNAS) INVOLVE IN THE EPIGENETIC REGULATION AND ITS MAJOR MECHANISM IS THE SEQUESTRATION OF THE TARGET MICRO RNAS (MIRNAS). WE HYPOTHESIZED THAT CIRCRNAS MIGHT BE RELATED WITH THE PATHOPHYSIOLOGY OF CHRONIC EPILEPSY AND EVALUATED THE ALTERED CIRCRNA EXPRESSIONS AND THEIR POSSIBLE REGULATORY EFFECTS ON THEIR TARGET MIRNAS AND MRNAS IN A MOUSE EPILEPSY MODEL. THE CIRCRNA EXPRESSION PROFILE IN THE HIPPOCAMPUS OF THE PILOCARPINE MICE WAS ANALYZED AND COMPARED WITH CONTROL. THE CORRELATION BETWEEN THE EXPRESSION OF MIRNA BINDING SITES (MIRNA RESPONSE ELEMENTS, MRE) IN THE DYSREGULATED CIRCRNAS AND THE EXPRESSION OF THEIR TARGET MIRNAS WAS EVALUATED. AS MIRNAS ALSO INHIBIT THEIR TARGET MRNAS, CIRCRNA-MIRNA-MRNA REGULATORY NETWORK, COMPRISED OF DYSREGULATED RNAS THAT TARGETS ONE ANOTHER WERE SEARCHED. FOR THE IDENTIFIED NETWORKS, BIOINFORMATICS ANALYSES WERE PERFORMED. AS THE RESULT, FORTY-THREE CIRCRNAS WERE DYSREGULATED IN THE HIPPOCAMPUS (UP-REGULATED, 26; DOWN-REGULATED, 17). THE CHANGE IN THE EXPRESSION OF MRE IN THOSE CIRCRNAS NEGATIVELY CORRELATED WITH THE CHANGE IN THE RELEVANT TARGET MIRNA EXPRESSION (R = -0.461, P<0.001), SUPPORTING THAT CIRCRNAS INHIBIT THEIR TARGET MIRNA. 333 DYSREGULATED CIRCRNA-MIRNA-MRNA NETWORKS WERE IDENTIFIED. GENE ONTOLOGY AND PATHWAY ANALYSES DEMONSTRATED THAT THE UP-REGULATED MRNAS IN THOSE NETWORKS WERE CLOSELY RELATED TO THE MAJOR PROCESSES IN EPILEPSY. AMONG THEM, STRING ANALYSIS IDENTIFIED 37 KEY MRNAS WITH ABUNDANT (>/=4) INTERACTIONS WITH OTHER DYSREGULATED TARGET MRNAS. THE DYSREGULATION OF THE CIRCRNAS WHICH HAD MULTIPLE INTERACTIONS WITH KEY MRNAS WERE VALIDATED BY PCR. WE CONCLUDED THAT DYSREGULATED CIRCRNAS MIGHT HAVE A PATHOPHYSIOLOGIC ROLE IN CHRONIC EPILEPSY BY REGULATING MULTIPLE DISEASE RELEVANT MRNAS VIA CIRCRNA-MIRNA-MRNA INTERACTIONS. 2018 19 1120 27 COMPARISON OF DIFFERENT HISTONE DEACETYLASE INHIBITORS IN ATTENUATING INFLAMMATORY PAIN IN RATS. HISTONE DEACETYLASE INHIBITORS (HDACIS), WHICH INTERFERE WITH THE EPIGENETIC PROCESS OF HISTONE ACETYLATION, HAVE SHOWN ANALGESIC EFFECTS IN ANIMAL MODELS OF PERSISTENT PAIN. THE HDAC FAMILY COMPRISES 18 GENES; HOWEVER, THE DIFFERENT EFFECTS OF DISTINCT CLASSES OF HDACIS ON PAIN RELIEF REMAIN UNCLEAR. THE AIM OF THIS STUDY WAS TO DETERMINE THE EFFICACY OF THESE HDACIS ON ATTENUATING THERMAL HYPERALGESIA IN PERSISTENT INFLAMMATORY PAIN. PERSISTENT INFLAMMATORY PAIN WAS INDUCED BY INJECTING COMPLETE FREUND'S ADJUVANT (CFA) INTO THE LEFT HIND PAW OF RATS. THEN, HDACIS TARGETING CLASS I (ENTINOSTAT (MS-275)) AND CLASS IIA (SODIUM BUTYRATE, VALPROIC ACID (VPA), AND 4-PHENYLBUTYRIC ACID (4-PBA)), OR CLASS II (SUBEROYLANILIDE HYDOXAMIC ACID (SAHA), TRICHOSTATIN A (TSA), AND DACINOSTAT (LAQ824)) WERE ADMINISTERED INTRAPERITONEALLY ONCE DAILY FOR 3 OR 4 DAYS. WE FOUND THAT THE INJECTION OF SAHA ONCE A DAY FOR 3 DAYS SIGNIFICANTLY ATTENUATED CFA-INDUCED THERMAL HYPERALGESIA FROM DAY 4 AND LASTED 7 DAYS. IN COMPARISON WITH SAHA, SUPPRESSION OF HYPERALGESIA BY 4-PBA PEAKED ON DAY 2, WHEREAS THAT BY MS-275 OCCURRED ON DAYS 5 AND 6. FATIGUE WAS A SERIOUS SIDE EFFECT SEEN WITH MS-275. THESE FINDINGS WILL BE BENEFICIAL FOR OPTIMIZING THE SELECTION OF SPECIFIC HDACIS IN MEDICAL FIELDS SUCH AS PAIN MEDICINE AND NEUROPSYCHIATRY. 2019 20 3192 29 HDAC INHIBITION COUNTERACTS METASTATIC RE-ACTIVATION OF PROSTATE CANCER CELLS INDUCED BY CHRONIC MTOR SUPPRESSION. THIS STUDY WAS DESIGNED TO INVESTIGATE WHETHER EPIGENETIC MODULATION BY HISTONE DEACETYLASE (HDAC) INHIBITION MIGHT CIRCUMVENT RESISTANCE TOWARDS THE MECHANISTIC TARGET OF RAPAMYCIN (MTOR) INHIBITOR TEMSIROLIMUS IN A PROSTATE CANCER CELL MODEL. PARENTAL (PAR) AND TEMSIROLIMUS-RESISTANT (RES) PC3 PROSTATE CANCER CELLS WERE EXPOSED TO THE HDAC INHIBITOR VALPROIC ACID (VPA), AND TUMOR CELL ADHESION, CHEMOTAXIS, MIGRATION, AND INVASION WERE EVALUATED. TEMSIROLIMUS RESISTANCE WAS CHARACTERIZED BY REDUCED BINDING OF PC3(RES) CELLS TO ENDOTHELIUM, IMMOBILIZED COLLAGEN, AND FIBRONECTIN, BUT INCREASED ADHESION TO LAMININ, AS COMPARED TO THE PARENTAL CELLS. CHEMOTAXIS, MIGRATION, AND INVASION OF PC3(RES) CELLS WERE ENHANCED FOLLOWING TEMSIROLIMUS RE-TREATMENT. INTEGRIN ALPHA AND BETA RECEPTORS WERE SIGNIFICANTLY ALTERED IN PC3(RES) COMPARED TO PC3(PAR) CELLS. VPA SIGNIFICANTLY COUNTERACTED TEMSIROLIMUS RESISTANCE BY DOWN-REGULATING TUMOR CELL(-)MATRIX INTERACTION, CHEMOTAXIS, AND MIGRATION. EVALUATION OF INTEGRIN EXPRESSION IN THE PRESENCE OF VPA REVEALED A SIGNIFICANT DOWN-REGULATION OF INTEGRIN ALPHA5 IN PC3(RES) CELLS. BLOCKING STUDIES DEMONSTRATED A CLOSE ASSOCIATION BETWEEN ALPHA5 EXPRESSION ON PC3(RES) AND CHEMOTAXIS. IN THIS IN VITRO MODEL, TEMSIROLIMUS RESISTANCE DROVE PROSTATE CANCER CELLS TO BECOME HIGHLY MOTILE, WHILE HDAC INHIBITION REVERSED THE METASTATIC ACTIVITY. THE VPA-INDUCED INHIBITION OF METASTATIC ACTIVITY WAS ACCOMPANIED BY A LOWERED INTEGRIN ALPHA5 SURFACE LEVEL ON THE TUMOR CELLS. 2018