1  1663 175 DOWNREGULATION OF DNA METHYLTRANSFERASE-3A AMELIORATES THE OSTEOGENIC DIFFERENTIATION ABILITY OF ADIPOSE-DERIVED STEM CELLS IN DIABETIC OSTEOPOROSIS VIA WNT/BETA-CATENIN SIGNALING PATHWAY. BACKGROUND: DIABETES-RELATED OSTEOPOROSIS (DOP) IS A CHRONIC DISEASE CAUSED BY THE HIGH GLUCOSE ENVIRONMENT THAT INDUCES A METABOLIC DISORDER OF OSTEOCYTES AND OSTEOBLAST-ASSOCIATED MESENCHYMAL STEM CELLS. THE PROCESSES OF BONE DEFECT REPAIR AND REGENERATION BECOME EXTREMELY DIFFICULT WITH DOP. ADIPOSE-DERIVED STEM CELLS (ASCS), AS SEED CELLS IN BONE TISSUE ENGINEERING TECHNOLOGY, PROVIDE A PROMISING THERAPEUTIC APPROACH FOR BONE REGENERATION IN DOP PATIENTS. THE OSTEOGENIC ABILITY OF ASCS IS LOWER IN A DOP MODEL THAN THAT OF CONTROL ASCS. DNA METHYLATION, AS A MECHANISM OF EPIGENETIC REGULATION, MAY BE INVOLVED IN DNA METHYLATION OF VARIOUS GENES, THEREBY PARTICIPATING IN BIOLOGICAL BEHAVIORS OF VARIOUS CELLS. EMERGING EVIDENCE SUGGESTS THAT INCREASED DNA METHYLATION LEVELS ARE ASSOCIATED WITH ACTIVATION OF WNT/BETA-CATENIN SIGNALING PATHWAY. THE PURPOSE OF THIS STUDY WAS TO INVESTIGATE THE INFLUENCE OF THE DIABETIC ENVIRONMENT ON THE OSTEOGENIC POTENTIAL OF ASCS, TO EXPLORE THE ROLE OF DNA METHYLATION ON OSTEOGENIC DIFFERENTIATION OF DOP-ASCS VIA WNT/BETA-CATENIN SIGNALING PATHWAY, AND TO IMPROVE THE OSTEOGENIC DIFFERENTIATION ABILITY OF ASCS WITH DOP. METHODS: DOP-ASCS AND CONTROL ASCS WERE ISOLATED FROM DOP C57BL/6 AND CONTROL MICE, RESPECTIVELY. THE MULTIPOTENCY OF DOP-ASCS WAS CONFIRMED BY ALIZARIN RED-S, OIL RED-O, AND ALCIAN BLUE STAINING. REAL-TIME POLYMERASE CHAIN REACTION (RT-PCR), IMMUNOFLUORESCENCE, AND WESTERN BLOTTING WERE USED TO ANALYZE CHANGES IN MARKERS OF OSTEOGENIC DIFFERENTIATION, DNA METHYLATION, AND WNT/BETA-CATENIN SIGNALING. ALIZARIN RED-S STAINING WAS ALSO USED TO CONFIRM CHANGES IN THE OSTEOGENIC ABILITY. DNMT SMALL INTERFERING RNA (SIRNA), SHRNA-DNMT3A, AND LVRNA-DNMT3A WERE USED TO ASSESS THE ROLE OF DNMT3A IN OSTEOGENIC DIFFERENTIATION OF CONTROL ASCS AND DOP-ASCS. MICRO-COMPUTED TOMOGRAPHY, HEMATOXYLIN AND EOSIN STAINING, AND MASSON STAINING WERE USED TO ANALYZE CHANGES IN THE OSTEOGENIC CAPABILITY WHILE DOWNREGULATING DNMT3A WITH LENTIVIRUS IN DOP MICE IN VIVO. RESULTS: THE PROLIFERATIVE ABILITY OF DOP-ASCS WAS LOWER THAN THAT OF CONTROL ASCS. DOP-ASCS SHOWED A DECREASE IN OSTEOGENIC DIFFERENTIATION CAPACITY, LOWER WNT/BETA-CATENIN SIGNALING PATHWAY ACTIVITY, AND A HIGHER LEVEL OF DNMT3A THAN CONTROL ASCS. WHEN DNMT3A WAS DOWNREGULATED BY SIRNA AND SHRNA, OSTEOGENIC-RELATED FACTORS RUNT-RELATED TRANSCRIPTION FACTOR 2 AND OSTEOPONTIN, AND ACTIVITY OF WNT/BETA-CATENIN SIGNALING PATHWAY WERE INCREASED, WHICH RESCUED THE POOR OSTEOGENIC POTENTIAL OF DOP-ASCS. WHEN DNMT3A WAS UPREGULATED BY LVRNA-DNMT3A, THE OSTEOGENIC ABILITY WAS INHIBITED. THE SAME RESULTS WERE OBTAINED IN VIVO. CONCLUSIONS: DNMT3A SILENCING RESCUES THE NEGATIVE EFFECTS OF DOP ON ASCS AND PROVIDES A POSSIBLE APPROACH FOR BONE TISSUE REGENERATION IN PATIENTS WITH DIABETIC OSTEOPOROSIS.	2022	

2  4850  27 OPIOIDS AND OPIOID RECEPTORS; UNDERSTANDING PHARMACOLOGICAL MECHANISMS AS A KEY TO THERAPEUTIC ADVANCES AND MITIGATION OF THE MISUSE CRISIS. OPIOIDS ARE A MAINSTAY IN ACUTE PAIN MANAGEMENT AND PRODUCE THEIR EFFECTS AND SIDE EFFECTS (E.G., TOLERANCE, OPIOID-USE DISORDER AND IMMUNE SUPPRESSION) BY INTERACTION WITH OPIOID RECEPTORS. I WILL DISCUSS OPIOID PHARMACOLOGY IN SOME CONTROVERSIAL AREAS OF ENQUIRY OF ANAESTHETIC RELEVANCE. THE MAIN OPIOID TARGET IS THE MICRO (MU,MOP) RECEPTOR BUT OTHER MEMBERS OF THE OPIOID RECEPTOR FAMILY, DELTA (DELTA; DOP) AND KAPPA (KAPPA; KOP) OPIOID RECEPTORS ALSO PRODUCE ANALGESIC ACTIONS. THESE ARE NALOXONE-SENSITIVE. THERE IS IMPORTANT CLINICAL DEVELOPMENT RELATING TO THE NOCICEPTIN/ORPHANIN FQ (NOP) RECEPTOR, AN OPIOID RECEPTOR THAT IS NOT NALOXONE-SENSITIVE. BETTER UNDERSTANDING OF THE DRIVERS FOR OPIOID EFFECTS AND SIDE EFFECTS MAY FACILITATE SEPARATION OF SIDE EFFECTS AND PRODUCTION OF SAFER DRUGS. OPIOIDS BIND TO THE RECEPTOR ORTHOSTERIC SITE TO PRODUCE THEIR EFFECTS AND CAN ENGAGE MONOMER OR HOMO-, HETERODIMER RECEPTORS. SOME LIGANDS CAN DRIVE ONE INTRACELLULAR PATHWAY OVER ANOTHER. THIS IS THE BASIS OF BIASED AGONISM (OR FUNCTIONAL SELECTIVITY). OPIOID ACTIONS AT THE ORTHOSTERIC SITE CAN BE MODULATED ALLOSTERICALLY AND POSITIVE ALLOSTERIC MODULATORS THAT ENHANCE OPIOID ACTION ARE IN DEVELOPMENT. AS WELL AS TARGETING LIGAND-RECEPTOR INTERACTION AND TRANSDUCTION, MODULATING RECEPTOR EXPRESSION AND HENCE FUNCTION IS ALSO TRACTABLE. THERE IS EVIDENCE FOR EPIGENETIC ASSOCIATIONS WITH DIFFERENT TYPES OF PAIN AND ALSO SUBSTANCE MISUSE. AS LONG AS THE OPIOID NARRATIVE IS DEFINED BY THE 'OPIOID CRISIS' THE DRIVE TO REMOVE THEM COULD GATHER PACE. THIS WILL DENY USE WHERE THEY ARE EFFECTIVE, AND ACCESS TO MORPHINE FOR PAIN RELIEF IN LOW INCOME COUNTRIES.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
3  5743  39 SMOKING SUPPRESSES THE THERAPEUTIC POTENTIAL OF ADIPOSE STEM CELLS IN CROHN'S DISEASE PATIENTS THROUGH EPIGENETIC CHANGES. PATIENTS WITH CROHN'S DISEASE (CD) WHO SMOKE ARE KNOWN TO HAVE A WORSE PROGNOSIS THAN NEVER-SMOKERS AND A HIGHER RISK FOR POST-SURGICAL RECURRENCE, WHEREAS PATIENTS WHO QUIT SMOKING AFTER SURGERY HAVE SIGNIFICANTLY LOWER POST-OPERATIVE RECURRENCE. THE HYPOTHESIS WAS THAT SMOKING INDUCES EPIGENETIC CHANGES THAT IMPAIR THE CAPACITY OF ADIPOSE STEM CELLS (ASCS) TO SUPPRESS THE IMMUNE SYSTEM. IT WAS ALSO QUESTIONED WHETHER THIS IMPAIRMENT REMAINS IN EX-SMOKERS WITH CD. ASCS WERE ISOLATED FROM NON-SMOKERS, SMOKERS AND EX-SMOKERS WITH CD AND THEIR INTERACTIONS WITH IMMUNE CELLS WERE STUDIED. THE ASCS FROM BOTH SMOKERS AND EX-SMOKERS PROMOTED MACROPHAGE POLARIZATION TO AN M1 PRO-INFLAMMATORY PHENOTYPE, WERE NOT ABLE TO INHIBIT T- AND B-CELL PROLIFERATION IN VITRO AND ENHANCED THE GENE AND PROTEIN EXPRESSION OF INFLAMMATORY MARKERS INCLUDING INTERLEUKIN-1B. GENOME-WIDE EPIGENETIC ANALYSIS USING TWO DIFFERENT BIOINFORMATIC APPROACHES REVEALED SIGNIFICANT CHANGES IN THE METHYLATION PATTERNS OF GENES THAT ARE CRITICAL FOR WOUND HEALING, IMMUNE AND METABOLIC RESPONSE AND P53-MEDIATED DNA DAMAGE RESPONSE IN ASCS FROM SMOKERS AND EX-SMOKERS WITH CD. IN CONCLUSION, CIGARETTE SMOKING INDUCES A PRO-INFLAMMATORY EPIGENETIC SIGNATURE IN ASCS THAT LIKELY COMPROMISES THEIR THERAPEUTIC POTENTIAL.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
4   364  53 AMELIORATION OF UREMIC TOXIN INDOXYL SULFATE-INDUCED OSTEOBLASTIC CALCIFICATION BY SET DOMAIN CONTAINING LYSINE METHYLTRANSFERASE 7/9 PROTEIN. BACKGROUND: VASCULAR CALCIFICATION (VC) IS A VERY COMMON PHENOMENON IN PATIENTS WITH CHRONIC KIDNEY DISEASE (CKD). IT HAS BEEN REPORTED THAT SOME HISTONE METHYLATION PLAY A ROLE IN VC AS AN EPIGENETIC REGULATOR. INDOXYL SULFATE (IS) IS A PROTEIN-BOUND UREMIC TOXIN THAT HAS BEEN PROVEN AS ONE OF THE MAJOR RISK FACTORS OF CARDIOVASCULAR DISEASE IN CKD. SET DOMAIN CONTAINING LYSINE METHYLTRANSFERASE 7/9 (SET7/9) IS ONE OF THE IMPORTANT HISTONE METHYLTRANSFERASES. OBJECTIVES: THIS STUDY AIMED TO DETERMINE THE EFFECT OF IS ON THE EXPRESSION OF SET7/9 AND THE ROLE OF SET7/9 IN IS-INDUCED OSTEOBLASTIC DIFFERENTIATION AND CALCIFICATION OF VASCULAR SMOOTH MUSCLE CELLS (VSMCS). METHODS: VSMCS WERE INCUBATED WITH VARIOUS CONCENTRATIONS OF IS FOR DIFFERENT DURATIONS TO ASSESS OSTEOBLASTIC DIFFERENTIATION AND EXPRESSION OF SET7/9. WESTERN BLOT ANALYSIS AND QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION WERE PERFORMED TO ASSESS THE PROTEIN AND MRNA LEVELS OF SET7/9 RESPECTIVELY. THE CALCIUM CONTENT WAS MEASURED TO EVALUATE CALCIFICATION. RESULTS: OSTEOBLASTIC DIFFERENTIATION AND CALCIFICATION OF VSMCS AND DOWNREGULATION OF THE EXPRESSION OF SET7/9 WERE OBSERVED AFTER IS TREATMENT. THE AUTOPHAGY WAS ACTIVATED AFTER TREATMENT WITH IS, WHEREAS THE INHIBITION OF THE AUTOPHAGY PARTIALLY ATTENUATED THE EFFECT OF IS ON BOTH THE STIMULATION OF THE EXPRESSION OF RUNT-RELATED TRANSCRIPTION FACTOR 2 AND CALCIUM DEPOSITION. CONCLUSIONS: OUR DATA DEMONSTRATED THAT SET7/9 DOWNREGULATION AND AUTOPHAGY ACTIVATION MAY BE THE KEY MECHANISM OF IS-INDUCED VC IN CKD.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
5  1632  43 DNMTS ARE INVOLVED IN TGF-BETA1-INDUCED EPITHELIAL-MESENCHYMAL TRANSITIONS IN AIRWAY EPITHELIAL CELLS. CHRONIC RHINOSINUSITIS (CRS) PATHOGENESIS IS CLOSELY RELATED TO TISSUE REMODELING, INCLUDING EPITHELIAL-MESENCHYMAL TRANSITION (EMT). EPIGENETIC MECHANISMS PLAY KEY ROLES IN EMT. DNA METHYLATION, MEDIATED BY DNA METHYLTRANSFERASES (DNMTS), IS AN EPIGENETIC MARKER THAT IS CRITICAL TO EMT. THE GOAL OF THIS STUDY WAS TO DETERMINE WHETHER DNMTS WERE INVOLVED IN TGF-BETA1-INDUCED EMT AND ELUCIDATE THE UNDERLYING MECHANISMS IN NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. GLOBAL DNA METHYLATION AND DNMT ACTIVITY WERE QUANTIFIED. DNMT EXPRESSION WAS MEASURED USING REAL-TIME PCR (QRT-PCR) IN HUMAN CRS TISSUES. MRNA AND PROTEIN LEVELS OF DNMTS, E-CADHERIN, VIMENTIN, ALPHA-SMA, AND FIBRONECTIN WERE DETERMINED USING RT-PCR AND WESTERN BLOTTING, RESPECTIVELY. DNMT1, DNMT3A, AND DNMT3B GENE EXPRESSION WERE KNOCKED DOWN USING SIRNA TRANSFECTION. MAPK PHOSPHORYLATION AND EMT-RELATED TRANSCRIPTION FACTOR LEVELS WERE DETERMINED USING WESTERN BLOTTING. SIGNALING PATHWAYS WERE ANALYZED USING SPECIFIC INHIBITORS OF MAPK. WE DEMONSTRATED THESE DATA IN PRIMARY NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. GLOBAL DNA METHYLATION, DNMT ACTIVITY, AND DNMT EXPRESSION INCREASED IN CRS TISSUES. DNMT EXPRESSION WAS POSITIVELY CORRELATED WITH LUND-MCKAY CT SCORES. TGF-BETA1 DOSE-DEPENDENTLY INDUCED DNMT EXPRESSION. FURTHER, 5-AZA INHIBITED TGF-BETA1-INDUCED DNMT, SNAIL, AND SLUG EXPRESSION RELATED TO EMT, AS WELL AS P38 AND JNK PHOSPHORYLATION IN A549 CELLS AND TGF-BETA1-INDUCED DNMT EXPRESSION AND EMT IN PRIMARY NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. TGF-BETA1-INDUCED DNMT EXPRESSION LEADS TO DNA METHYLATION AND EMT VIA P38, JNK, SNAIL, AND SLUG SIGNALING PATHWAYS. INHIBITION OF DNMT SUPPRESSED THE EMT PROCESS AND THEREFORE IS POTENTIALLY A CRS THERAPEUTIC STRATEGY.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
6  4303  48 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
7  5868  37 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
8  2941  35 GENETIC AND EPIGENETIC ALTERATIONS IN TOLL LIKE RECEPTOR 2 AND WOUND HEALING IMPAIRMENT IN TYPE 2 DIABETES PATIENTS. AIM: PERSISTENT HYPERGLYCEMIC MICROENVIRONMENT IN TYPE 2 DIABETES MELLITUS (T2DM) LEADS TO THE DEVELOPMENT OF SECONDARY COMPLICATIONS LIKE WOUND HEALING IMPAIRMENT. PROPER CO-ORDINATION OF INNATE IMMUNE SYSTEM PLAYS AN INTEGRAL ROLE IN WOUND HEALING. TOLL LIKE RECEPTORS (TLRS) ARE PROMINENT CONTRIBUTORS FOR THE INDUCTION OF THE INNATE IMMUNE AND INFLAMMATION RESPONSE. TLR2 IS AN IMPORTANT EXTRACELLULAR MEMBER IN MAMMALIAN TLR FAMILY AND HAS BEEN SHOWN TO BE A POTENT PLAYER IN THE WOUND HEALING MECHANISM. METHODS: EXPRESSIONAL STATUS OF TLR2 WAS SEEN IN WOUNDS OF T2DM CASES WITH RESPECT TO THE SEVERITY OF WOUNDS IN 110 HUMAN LOWER EXTREMITY WOUNDS. THE METHYLATION STATUS OF TLR2 PROMOTER WAS ALSO EXAMINED. RESULTS: ALTHOUGH TLR2 TRANSCRIPTS WERE DOWNREGULATED IN T2DM WOUNDS COMPARED TO CONTROL, THEIR LEVELS TEND TO INCREASE WITH THE SEVERITY OF T2DM WOUNDS. THE METHYLATION STATUS OF TLR2 GENE PROMOTER WAS NOT SIGNIFICANTLY DIFFERENT AMONG DIFFERENT GRADES OF WOUNDS IN T2DM SUBJECTS. THE CPG SITES INVESTIGATED WERE TOTALLY OR PARTIALLY METHYLATED IN MAJORITY OF DFU CASES. CONCLUSION: TLR2 DOWN REGULATION IN WOUNDS OF T2DM PATIENTS COMPARED TO NON DIABETIC PATIENTS MAY LEAD TO DEVELOPMENT OF NON HEALING CHRONIC ULCERS IN THEM.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
9  5479  44 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
10  2326  50 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
11  5129  36 POSTOPERATIVE PAIN AND ANALGESIA: IS THERE A GENETIC BASIS TO THE OPIOID CRISIS? BACKGROUND: MULTIPLE FACTORS HAVE BEEN IMPLICATED IN DETERMINING WHY CERTAIN PATIENTS HAVE INCREASED POSTOPERATIVE PAIN, WITH THE POTENTIAL TO DEVELOP CHRONIC PAIN. THE PURPOSE OF THIS STUDY WAS TO: 1) IDENTIFY AND DESCRIBE GENES THAT AFFECT POSTOPERATIVE PAIN PERCEPTION AND CONTROL; 2) ADDRESS MODIFIABLE RISK FACTORS THAT RESULT IN EPIGENETIC ALTERED RESPONSES TO PAIN; AND 3) CHARACTERIZE DIFFERENCES IN PAIN SENSITIVITY AND THRESHOLDS BETWEEN OPIOID-NAIVE AND OPIOID-DEPENDENT PATIENTS. MATERIALS AND METHODS: THREE ELECTRONIC DATABASES WERE USED TO CONDUCT THE LITERATURE SEARCH: PUBMED, EBSCO HOST, AND SCOPUS. A TOTAL OF 372 ABSTRACTS WERE REVIEWED, OF WHICH 46 STUDIES WERE DEEMED RELEVANT AND ARE INCLUDED IN THIS REVIEW. RESULTS: SPECIFIC GENE ALTERATIONS THAT WERE SHOWN TO AFFECT POSTOPERATIVE PAIN CONTROL INCLUDED SINGLE NUCLEOTIDE POLYMORPHISMS IN THE MU, KAPPA, AND DELTA OPIOID RECEPTORS, ION CHANNEL GENES, CYTOTOXIC T-CELLS, GLUTAMATE RECEPTORS AND CYTOKINE GENES, AMONG OTHERS. ALCOHOLISM, OBESITY, AND SMOKING WERE ALL LINKED WITH GENETIC POLYMORPHISMS THAT ALTERED PAIN SENSITIVITY. OPIOID ABUSE WAS FOUND TO BE ASSOCIATED WITH A POORER RESPONSE TO ANALGESICS POSTOPERATIVELY, AS WELL AS A RISK FOR PRESCRIPTION OVERDOSE. CONCLUSION: ALTHOUGH PAIN PERCEPTION HAS MULTIPLE COMPLEX INFLUENCES, THE GREATEST VARIABILITY SEEN IN RESPONSE TO OPIOIDS AMONG POSTOPERATIVE PATIENTS KNOWN TO DATE CAN BE TRACED TO GENETIC DIFFERENCES IN OPIOID METABOLISM. FURTHER STUDY IS NEEDED TO DETERMINE THE CLINICAL SIGNIFICANCE OF THESE GENETIC ASSOCIATIONS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
12  1826  45 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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
13  6658  48 UPREGULATED LNCRNA H19 SPONGES MIR-106A-5P AND CONTRIBUTES TO ALDOSTERONE-INDUCED VASCULAR CALCIFICATION VIA ACTIVATING THE RUNX2-DEPENDENT PATHWAY. BACKGROUND: EXCESS ALDOSTERONE IS IMPLICATED IN VASCULAR CALCIFICATION (VC), BUT THE MECHANISM BY WHICH ALDOSTERONE-MR (MINERALOCORTICOID RECEPTOR) COMPLEX PROMOTES VC IS UNCLEAR. EMERGING EVIDENCE INDICATES THAT LONG-NONCODING RNA H19 (H19) PLAYS A CRITICAL ROLE IN VC. WE EXAMINED WHETHER ALDOSTERONE-INDUCED OSTEOGENIC DIFFERENTIATION OF VASCULAR SMOOTH MUSCLE CELLS (VSMCS) THROUGH H19 EPIGENETIC MODIFICATION OF RUNX2 (RUNT-RELATED TRANSCRIPTION FACTOR-2) IN A MR-DEPENDENT MANNER. METHODS: WE INDUCED IN VIVO RAT MODEL OF CHRONIC KIDNEY DISEASE USING A HIGH ADENINE AND PHOSPHATE DIET TO EXPLORE THE RELATIONSHIP AMONG ALDOSTERONE, MR, H19, AND VC. WE ALSO CULTURED HUMAN AORTIC VSMCS TO EXPLORE THE ROLES OF H19 IN ALDOSTERONE-MR COMPLEX-INDUCED OSTEOGENIC DIFFERENTIATION AND CALCIFICATION OF VSMCS. RESULTS: H19 AND RUNX2 WERE SIGNIFICANTLY INCREASED IN ALDOSTERONE-INDUCED VSMC OSTEOGENIC DIFFERENTIATION AND VC, BOTH IN VITRO AND IN VIVO, WHICH WERE SIGNIFICANTLY BLOCKED BY THE MR ANTAGONIST SPIRONOLACTONE. MECHANISTICALLY, OUR FINDINGS REVEAL THAT THE ALDOSTERONE-ACTIVATED MR BOUND TO H19 PROMOTER AND INCREASED ITS TRANSCRIPTIONAL ACTIVITY, AS DETERMINED BY CHROMATIN IMMUNOPRECIPITATION, ELECTROPHORETIC MOBILITY SHIFT ASSAY, AND LUCIFERASE REPORTER ASSAY. SILENCING H19 INCREASED MICRORNA-106A-5P (MIR-106A-5P) EXPRESSION, WHICH SUBSEQUENTLY INHIBITED ALDOSTERONE-INDUCED RUNX2 EXPRESSION AT THE POSTTRANSCRIPTIONAL LEVEL. IMPORTANTLY, WE OBSERVED A DIRECT INTERACTION BETWEEN H19 AND MIR-106A-5P, AND DOWNREGULATION OF MIR-106A-5P EFFICIENTLY REVERSED THE SUPPRESSION OF RUNX2 INDUCED BY H19 SILENCING. CONCLUSIONS: OUR STUDY CLARIFIES A NOVEL MECHANISM BY WHICH UPREGULATION OF H19 CONTRIBUTES TO ALDOSTERONE-MR COMPLEX-PROMOTED RUNX2-DEPENDENT VSMC OSTEOGENIC DIFFERENTIATION AND VC THROUGH SPONGING MIR-106A-5P. THESE FINDINGS HIGHLIGHT A POTENTIAL THERAPEUTIC TARGET FOR ALDOSTERONE-INDUCED VC.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
14  1631  52 DNMT3A METHYLATION IN NEUROPATHIC PAIN. BACKGROUND: MU OPIOID RECEPTOR (MOR) PLAYS A CRUCIAL ROLE IN MEDIATING ANALGESIC EFFECTS OF OPIOIDS AND IS CLOSELY ASSOCIATED WITH THE PATHOLOGIES OF NEUROPATHIC PAIN. PREVIOUS STUDIES HAVE REPORTED THAT PERIPHERAL NERVE INJURY DOWNREGULATES MOR EXPRESSION, BUT THE EPIGENETIC MECHANISMS REMAIN UNKNOWN. OBJECTIVE: THEREFORE, WE INVESTIGATED DNA METHYLTRANSFERASE3A (DNMT3A) EXPRESSION OR METHYLATION CHANGES WITHIN MOR PROMOTER IN THE SPINAL CORD IN A NEUROPATHIC PAIN INDUCED BY A CHRONIC CONSTRICTION INJURY (CCI) MOUSE MODEL AND FURTHER DETERMINED WHETHER THESE INJURY-ASSOCIATED CHANGES ARE REVERSIBLE BY PHARMACOLOGICAL INTERVENTIONS. METHODS: A CCI MOUSE MODEL WAS ESTABLISHED AND TISSUE SPECIMENS OF LUMBAR SPINAL CORDS WERE COLLECTED. THE NOCICEPTION THRESHOLD WAS EVALUATED BY A MODEL HEATED 400 BASE. DNMT3A AND MOR MRNA AND PROTEIN LEVEL WERE DETECTED BY REAL-TIME-POLYMERASE CHAIN REACTION AND WESTERN BLOT, RESPECTIVELY. METHYLATION OF DNMT3A GENE WAS MEASURED BY METHYLATION-SPECIFIC PCR. RESULTS: OUR DATA SHOWED THAT CHRONIC NERVE INJURY LED TO A SIGNIFICANT UPREGULATION OF DNMT3A EXPRESSION THAT WAS ASSOCIATED WITH INCREASED METHYLATION OF MOR GENE PROMOTER AND DECREASED MOR PROTEIN EXPRESSION IN THE SPINAL CORD. INHIBITION OF DNMT3A CATALYTIC ACTIVITY WITH DNMT INHIBITOR RG108 SIGNIFICANTLY BLOCKED THE INCREASE IN METHYLATION OF THE MOR PROMOTER, AND THEN UPREGULATED MOR EXPRESSION AND ATTENUATED THERMAL HYPERALGESIA IN NEUROPATHIC PAIN MICE. CONCLUSION: THIS STUDY DEMONSTRATES THAT AN INCREASE OF DNMT3A EXPRESSION AND MOR METHYLATION EPIGENETICALLY PLAY AN IMPORTANT ROLE IN NEUROPATHIC PAIN. TARGETING DNMT3A TO THE PROMOTER OF MOR GENE BY DNMT INHIBITOR MAY BE A PROMISING APPROACH TO THE DEVELOPMENT OF NEW NEUROPATHIC PAIN THERAPY.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
15  6216  33 THE INVOLVEMENT OF COPPER, CIRCULAR RNAS, AND INFLAMMATORY CYTOKINES IN CHRONIC RESPIRATORY DISEASE. EXPOSURE TO HIGH CONCENTRATIONS OF COPPER IS ASSOCIATED WITH PULMONARY INFLAMMATION AND CHRONIC RESPIRATORY DISEASE (CRD). EPIGENETIC MODULATION OF NONCODING RNAS CONTRIBUTES TO THE DEVELOPMENT OF SEVERAL CRDS. IT IS UNKNOWN WHETHER EPIGENETIC MODULATION IS INVOLVED IN COPPER MEDIATED PULMONARY INFLAMMATION AND CRD. WE CONDUCTED A CASE-CONTROL STUDY OF 101 CRD CASES AND 161 CONTROL SUBJECTS IN SHIJIAZHUANG, CHINA, AND EVALUATED CIRCRNAS AND CYTOKINE LEVELS (IL-6 AND IL-8) BY QPCR AND ELISA. URINARY COPPER CONCENTRATION WAS DETERMINED BY INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY. LINEAR MIXED MODELS AND GENERALIZED LINEAR MIXED MODELS WERE USED TO ASSESS THE ASSOCIATIONS OF CIRCRNAS WITH CRD, URINARY COPPER, AND CYTOKINES. WE EXPOSED THE HUMAN BRONCHIAL EPITHELIAL CELL LINE, 16HBE, TO COPPER AND ASSESSED THE FUNCTIONAL ROLE OF A CIRCRNA, CIRC_0008882, BY RNA OVEREXPRESSION. CELLULAR LOCATION OF CIRC_0008882 WAS ASSESSED BY SEPARATION OF NUCLEAR AND CYTOPLASMIC RNAS. NINE CIRCRNAS WERE ASSOCIATED WITH AN INCREASED RISK FOR CRDS, WHILE THE RELATIVE EXPRESSION OF CIRC_0008882 WAS DECREASED AFTER COPPER EXPOSURE IN VITRO AND IN VIVO. COPPER EXPOSURE STIMULATED 16HBE CELLS TO RELEASE PROINFLAMMATORY IL-6 AND IL-8. THE RELEASE OF THE CYTOKINES WAS INHIBITED BY OVEREXPRESSION OF CIRC_0008882. THESE RESULTS SUGGEST A ROLE FOR CIRC_0008882 IN THE REGULATION OF CRD ASSOCIATED INFLAMMATION FOLLOWING COPPER EXPOSURE.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
16  1158  52 CONTEXT-DEPENDENT EPIGENETIC REGULATION OF NUCLEAR FACTOR OF ACTIVATED T CELLS 1 IN PANCREATIC PLASTICITY. BACKGROUND & AIMS: THE ABILITY OF EXOCRINE PANCREATIC CELLS TO CHANGE THE CELLULAR PHENOTYPE IS REQUIRED FOR TISSUE REGENERATION UPON INJURY, BUT ALSO CONTRIBUTES TO THEIR MALIGNANT TRANSFORMATION AND TUMOR PROGRESSION. WE INVESTIGATED CONTEXT-DEPENDENT SIGNALING AND TRANSCRIPTION MECHANISMS THAT DETERMINE PANCREATIC CELL FATE DECISIONS TOWARD REGENERATION AND MALIGNANCY. IN PARTICULAR, WE STUDIED THE FUNCTION AND REGULATION OF THE INFLAMMATORY TRANSCRIPTION FACTOR NUCLEAR FACTOR OF ACTIVATED T CELLS 1 (NFATC1) IN PANCREATIC CELL PLASTICITY AND TISSUE ADAPTATION. METHODS: WE ANALYZED CELL PLASTICITY DURING PANCREATIC REGENERATION AND TRANSFORMATION IN MICE WITH PANCREAS-SPECIFIC EXPRESSION OF A CONSTITUTIVELY ACTIVE FORM OF NFATC1, OR DEPLETION OF ENHANCER OF ZESTE 2 HOMOLOGUE 2 (EZH2), IN THE CONTEXT OF WILD-TYPE OR CONSTITUTIVELY ACTIVATE KRAS, RESPECTIVELY. ACUTE AND CHRONIC PANCREATITIS WERE INDUCED BY INTRAPERITONEAL INJECTION OF CAERULEIN. EZH2-DEPENDENT REGULATION OF NFATC1 EXPRESSION WAS STUDIED IN MOUSE IN HUMAN PANCREATIC TISSUE AND CELLS BY IMMUNOHISTOCHEMISTRY, IMMUNOBLOTTING, AND QUANTITATIVE REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION. WE USED GENETIC AND PHARMACOLOGIC APPROACHES OF EZH2 AND NFATC1 INHIBITION TO STUDY THE CONSEQUENCES OF PATHWAY DISRUPTION ON PANCREATIC MORPHOLOGY AND FUNCTION. EPIGENETIC MODIFICATIONS ON THE NFATC1 GENE WERE INVESTIGATED BY CHROMATIN IMMUNOPRECIPITATION ASSAYS. RESULTS: NFATC1 WAS RAPIDLY AND TRANSIENTLY INDUCED IN EARLY ADAPTATION TO ACINAR CELL INJURY IN HUMAN SAMPLES AND IN MICE, WHERE IT PROMOTED ACINAR CELL TRANSDIFFERENTIATION AND BLOCKED PROLIFERATION OF METAPLASTIC PANCREATIC CELLS. HOWEVER, IN LATE STAGES OF REGENERATION, NFATC1 WAS EPIGENETICALLY SILENCED BY EZH2-DEPENDENT HISTONE METHYLATION, TO ENABLE ACINAR CELL REDIFFERENTIATION AND PREVENT ORGAN ATROPHY AND EXOCRINE INSUFFICIENCY. IN CONTRAST, ONCOGENIC ACTIVATION OF KRAS SIGNALING IN PANCREATIC DUCTAL ADENOCARCINOMA CELLS REVERSED THE EZH2-DEPENDENT EFFECTS ON THE NFATC1 GENE AND WAS REQUIRED FOR EZH2-MEDIATED TRANSCRIPTIONAL ACTIVATION OF NFATC1. CONCLUSIONS: IN STUDIES OF HUMAN AND MOUSE PANCREATIC CELLS AND TISSUE, WE IDENTIFIED CONTEXT-SPECIFIC EPIGENETIC REGULATION OF NFATC1 ACTIVITY AS AN IMPORTANT MECHANISM OF PANCREATIC CELL PLASTICITY. INHIBITORS OF EZH2 MIGHT THEREFORE INTERFERE WITH ONCOGENIC ACTIVITY OF NFATC1 AND BE USED IN TREATMENT OF PANCREATIC DUCTAL ADENOCARCINOMA.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
17  5034  34 PHARMACOEPIGENETICS OF THE ROLE OF DNA METHYLATION IN MU-OPIOID RECEPTOR EXPRESSION IN DIFFERENT HUMAN BRAIN REGIONS. AIM: EXPOSURE TO OPIOIDS HAS BEEN ASSOCIATED WITH EPIGENETIC EFFECTS. STUDIES IN RODENTS SUGGESTED A ROLE OF VARYING DEGREES OF DNA METHYLATION IN THE DIFFERENTIAL REGULATION OF MU-OPIOID RECEPTOR EXPRESSION ACROSS THE BRAIN. METHODS: IN A TRANSLATIONAL INVESTIGATION, USING TISSUE ACQUIRED POSTMORTEM FROM 21 BRAIN REGIONS OF FORMER OPIATE ADDICTS, REPRESENTING A HUMAN COHORT WITH CHRONIC OPIOID EXPOSURE, MU-OPIOID RECEPTOR EXPRESSION WAS ANALYZED AT THE LEVEL OF DNA METHYLATION, MRNA AND PROTEIN. RESULTS & CONCLUSION: WHILE HIGH OR LOW MU-OPIOID RECEPTOR EXPRESSION SIGNIFICANTLY CORRELATED WITH LOCAL OPRM1 MRNA LEVELS, THERE WAS NO CORRESPONDING ASSOCIATION WITH OPRM1 METHYLATION STATUS. ADDITIONAL EXPERIMENTS IN HUMAN CELL LINES SHOWED THAT CHANGES IN DNA METHYLATION ASSOCIATED WITH CHANGES IN MU-OPIOID EXPRESSION WERE AN ORDER OF MAGNITUDE GREATER THAN DIFFERENCES IN BRAIN. HENCE, DIFFERENT DEGREES OF DNA METHYLATION ASSOCIATED WITH CHRONIC OPIOID EXPOSURE ARE UNLIKELY TO EXERT A MAJOR ROLE IN THE REGION-SPECIFICITY OF MU-OPIOID RECEPTOR EXPRESSION IN THE HUMAN BRAIN.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
18  5568  46 ROLE OF MARIJUANA COMPONENTS ON THE REGENERATIVE ABILITY OF STEM CELLS. STEM CELL THERAPY PROMOTES TISSUE REGENERATION AND WOUND HEALING. EFFORTS HAVE BEEN MADE TO PRIME STEM CELLS TO ENHANCE THEIR REGENERATIVE ABILITIES. CERTAIN MARIJUANA COMPONENTS, NAMELY THE NON-PSYCHOACTIVE CANNABIDIOL (CBD) AND PSYCHOACTIVE TETRAHYDROCANNABINOL (THC), ARE DEFINED AS IMMUNOMODULATORS.(9) WE TEST WHETHER TWO SOURCES OF STEM CELLS, PRIMED WITH CBD OR THC, WOULD DEMONSTRATE IMPROVED REGENERATIVE ABILITIES. HUMAN ADIPOSE-DERIVED STEM CELLS (ASCS) AND BONE MARROW-DERIVED STEM CELLS (BMDSCS), NOT OBTAINED FROM THE SAME INDIVIDUAL, WERE TREATED WITH LOW (300 NM) OR HIGH (3 MUM) CONCENTRATION CBD. PORCINE ASCS AND BMDSCS WERE ISOLATED FROM A SINGLE PIG, AND TREATED WITH EITHER LOW OR HIGH CONCENTRATIONS OF CBD OR THC. TRANSWELL MIGRATION AND MTT PROLIFERATION ASSAYS WERE PERFORMED ON THE HUMAN ASCS AND BMDSCS. ALSO, TRANSWELL MIGRATION ASSAY WAS PERFORMED ON THE PORCINE ASCS AND BMDSCS. FINALLY, A WOUND HEALING SCRATCH ASSAY IN PORCINE PRIMARY FIBROBLASTS (PFS) WAS PERFORMED, CO-CULTURED WITH THE CANNABINOID-TREATED ASCS. CBD PRIMING AT LOW CONCENTRATION INDUCES MIGRATION BY 180% (P < .01) IN PORCINE ASCS, AND BY ONLY 93% (P < .02) IN PORCINE BMDSCS. IN PORCINE STEM CELLS, THC PRIMING AT LOW CONCENTRATION INDUCES MIGRATION BY 91.6% (P < .01) IN ASCS BUT BY ONLY 44.3% (P < .03) IN BMDSCS. COMPARED TO PFS CO-CULTURED WITH UNTREATED ASCS, PFS CO-CULTURED WITH LOW CBD-PRIMED ASCS HAD 75% FASTER WOUND CLOSURE AT 18 HOURS (P < .01). CBD AND THC PRIMING OF ASCS AND BMDSCS, PARTICULARLY AT LOWER DOSES, ENHANCES A NUMBER OF REGENERATIVE PARAMETERS, SUGGESTING THAT THESE MAJOR MARIJUANA COMPONENTS MAY IMPROVE STEM CELL-BASED THERAPIES. SIGNIFICANCE OF THE STUDY: OUR STUDY DEMONSTRATES THAT CANNABINOIDS CAN ENHANCE THE REGENERATIVE CAPACITY OF TWO MAJOR SOURCES OF STEM CELLS, ADIPOSE- AND BONE MARROW-DERIVED, FROM HUMAN AND PORCINE DONORS. STEM CELL ISOLATION AND EXPANSION IS INVASIVE, COSTLY AND TIME CONSUMING. STEM CELLS WITH IMPROVED REGENERATIVE PROPERTIES MAY BE EFFECTIVE IN THE TREATMENT OF ACUTE OR CHRONIC WOUNDS. THIS IS THE FIRST STUDY TO COMPARE THE PRIMING POTENTIAL OF TWO SOURCES OF STEM CELLS FROM THE SAME ANIMAL, WITH THE SAME GENETIC AND EPIGENETIC PROFILE, AS WELL AS THE FIRST TO PRIME WITH THC.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
19  3809  45 INTRAPERITONEAL 5-AZACYTIDINE ALLEVIATES NERVE INJURY-INDUCED PAIN IN RATS BY MODULATING DNA METHYLATION. TO INVESTIGATE THE ROLE OF DNA METHYLATION IN MODULATING CHRONIC NEUROPATHIC PAIN (NPP), IDENTIFY POSSIBLE TARGET GENES OF DNA METHYLATION INVOLVED IN THIS PROCESS, AND PRELIMINARILY CONFIRM THE MEDICINAL VALUE OF THE DNA METHYLTRANSFERASES (DNMTS) INHIBITOR 5-AZACYTIDINE (5-AZA) IN NPP BY TARGETING GENE METHYLATION. TWO RAT NPP MODELS, CHRONIC CONSTRICTION INJURY (CCI) AND SPINAL NERVE LIGATION (SNL), WERE USED. THE DNA METHYLATION PROFILES IN THE LUMBAR SPINAL CORD WERE ASSAYED USING AN ARRAYSTAR RAT REFSEQ PROMOTER ARRAY. THE UNDERLYING GENES WITH DIFFERENTIAL METHYLATION WERE THEN IDENTIFIED AND SUBMITTED TO GENE ONTOLOGY AND PATHWAY ANALYSIS. METHYL-DNA IMMUNOPRECIPITATION QUANTITATIVE PCR (MEDIP-QPCR) AND QUANTITATIVE REVERSE TRANSCRIPTION-PCR (RT-QPCR) WERE USED TO CONFIRM GENE METHYLATION AND EXPRESSION. THE PROTECTIVE FUNCTION OF 5-AZA IN NPP AND GENE EXPRESSION WERE EVALUATED VIA BEHAVIORAL ASSAYS AND RT-QPCR, RESPECTIVELY. ANALYSIS OF THE DNA METHYLATION PATTERNS IN THE LUMBAR SPINAL CORD INDICATED THAT 1205 DIFFERENTIALLY METHYLATED FRAGMENTS IN CCI RATS WERE LOCATED WITHIN DNA PROMOTER REGIONS, INCLUDING 638 HYPERMETHYLATED FRAGMENTS AND 567 HYPOMETHYLATED FRAGMENTS. THE METHYLATION LEVELS OF GRM4, HTR4, ADRB2, KCNF1, GAD2, AND PPARG, WHICH ARE ASSOCIATED WITH LONG-TERM POTENTIATION (LTP) AND GLUTAMATERGIC SYNAPSE PATHWAYS, WERE INCREASED WITH A CORRESPONDING DECREASE IN THEIR MRNA EXPRESSION, IN THE SPINAL CORDS OF CCI RATS. MOREOVER, WE FOUND THAT THE INTRAPERITONEAL INJECTION OF 5-AZA (4 MG/KG) ATTENUATED CCI- OR SNL-INDUCED MECHANICAL ALLODYNIA AND THERMAL HYPERALGESIA. FINALLY, THE MRNA EXPRESSION OF HYPERMETHYLATED GENES SUCH AS GRM4, HTR4, ADRB2, KCNF1, AND GAD2 WAS REVERSED AFTER 5-AZA TREATMENT. CCI INDUCED WIDESPREAD METHYLATION CHANGES IN THE DNA PROMOTER REGIONS IN THE LUMBAR SPINAL CORD. INTRAPERITONEAL 5-AZA ALLEVIATED HYPERALGESIA IN CCI AND SNL RATS, AN EFFECT ACCOMPANIED BY THE REVERSED EXPRESSION OF HYPERMETHYLATED GENES. THUS, DNA METHYLATION INHIBITION REPRESENTS A PROMISING EPIGENETIC STRATEGY FOR PROTECTION AGAINST CHRONIC NPP FOLLOWING NERVE INJURY. OUR STUDY LAYS A THEORETICAL FOUNDATION FOR 5-AZA TO BECOME A CLINICAL TARGETED DRUG.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
20  4854  28 OPRM1 METHYLATION CONTRIBUTES TO OPIOID TOLERANCE IN CANCER PATIENTS. CANCER PATIENTS IN PAIN REQUIRE HIGH DOSES OF OPIOIDS AND QUICKLY BECOME OPIOID-TOLERANT. PREVIOUS STUDIES HAVE SHOWN THAT CHRONIC CANCER PAIN AS WELL AS HIGH-DOSE OPIOID USE LEAD TO MU-OPIOID RECEPTOR DOWNREGULATION. IN THIS STUDY WE EXPLORE DOWNREGULATION OF THE MU-OPIOID RECEPTOR GENE (OPRM1), AS A MECHANISM FOR OPIOID TOLERANCE IN THE SETTING OF OPIOID USE FOR CANCER PAIN. WE DEMONSTRATE IN A COHORT OF 84 CANCER PATIENTS THAT HIGH-DOSE OPIOID USE CORRELATES WITH OPRM1 HYPERMETHYLATION IN PERIPHERAL LEUKOCYTES OF THESE PATIENTS. WE THEN REVERSE-TRANSLATE OUR CLINICAL FINDINGS BY CREATING A MOUSE CANCER PAIN MODEL; WE CREATE OPIOID TOLERANCE IN THE MOUSE CANCER MODEL TO MIMIC OPIOID TOLERANCE IN THE CANCER PATIENTS. USING THIS MODEL WE DETERMINE THE FUNCTIONAL SIGNIFICANCE OF OPRM1 METHYLATION ON CANCER PAIN AND OPIOID TOLERANCE. WE FOCUS ON 2 MAIN CELLS WITHIN THE CANCER MICROENVIRONMENT: THE CANCER CELL AND THE NEURON. WE SHOW THAT TARGETED RE-EXPRESSION OF MU-OPIOID RECEPTOR ON CANCER CELLS INHIBITS MECHANICAL AND THERMAL HYPERSENSITIVITY, AND PREVENTS OPIOID TOLERANCE, IN THE MOUSE MODEL. THE RESULTANT ANALGESIA AND PROTECTION AGAINST OPIOID TOLERANCE ARE LIKELY DUE TO PRESERVATION OF MU-OPIOID RECEPTOR EXPRESSION ON THE CANCER-ASSOCIATED NEURONS. PERSPECTIVE: WE DEMONSTRATE THAT EPIGENETIC REGULATION OF OPRM1 CONTRIBUTES TO OPIOID TOLERANCE IN CANCER PATIENTS, AND THAT TARGETED GENE THERAPY COULD TREAT CANCER-INDUCED NOCICEPTION AND OPIOID TOLERANCE IN A MOUSE CANCER MODEL.	2017