1  2863 120 FUNCTION OF DNA METHYLTRANSFERASE 3A IN LEAD (PB(2+) )-INDUCED CYCLOOXYGENASE-2 GENE. LEAD IONS (PB(2+) ) ARE TOXIC INDUSTRIAL POLLUTANTS ASSOCIATED WITH CHRONIC INFLAMMATORY DISEASES IN HUMANS AND ANIMALS. PREVIOUSLY, WE FOUND THAT PB(2+) IONS INDUCE COX-2 GENE EXPRESSION VIA THE EGF RECEPTOR/NUCLEAR FACTOR-KAPPAB SIGNAL TRANSDUCTION PATHWAY IN EPIDERMOID CARCINOMA CELL LINE A431. IN THIS STUDY, TO SEE WHETHER PB(2+) IONS AFFECT COX-2 EXPRESSION BY EPIGENETIC MECHANISMS, WE LOOKED AT THE MRNAS OF DNA METHYLTRANSFERASES (DNMTS) USING REAL-TIME PCR OF TOTAL RNA FROM THESE CELLS. CELLS EXPOSED TO PB(2+) HAD LOW LEVELS OF DNMT3A MRNA, WHEREAS THE LEVELS OF DNMT1 AND DNMT3B MRNAS REMAINED UNCHANGED. PRETREATMENT OF CELLS WITH DNMT INHIBITOR 5-AZA-2'-DEOXYCYTIDINE (5 MUM) FOLLOWED BY PB(2+) (1 MUM) SIGNIFICANTLY INCREASED LEVELS OF COX-2 MRNA COMPARED WITH CELLS TREATED WITH PB(2+) ALONE. OVEREXPRESSION OF TUMOR SUPPRESSOR GENE RB CORRELATED WITH AN INCREASE IN COX-2 MRNA AND A DECREASE IN DNMT3A MRNA. CONVERSELY, OVEREXPRESSION OF TRANSCRIPTION FACTOR E2F1 CORRELATED WITH A DECREASE IN COX-2 MRNA AND AN INCREASE IN DMNT3A MRNA. PRETREATMENT WITH EGFR INHIBITORS AG1478 AND PD153035 SIGNIFICANTLY LIMITED PB(2+) -INDUCED REDUCTION IN DNMT3A MRNA. IN ADDITION, GENE KNOCKDOWN OF DNMT3A WITH SHORT HAIRPIN RNA CORRELATED WITH INCREASED COX-2 MRNA INDUCED BY PB(2+) . OUR FINDINGS SUGGEST PB(2+) IONS INDUCE COX-2 EXPRESSION INDIRECTLY BY REDUCING DNMT3A METHYLATION OF THE COX-2 PROMOTER VIA TRANSCRIPTION FACTORS RB AND E2F1.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
2  6566  24 TRANSLATIONAL PERSPECTIVES TO TREAT EPIDERMOLYSIS BULLOSA-WHERE DO WE STAND? EPIDERMOLYSIS BULLOSA (EB) IS THE PROTOTYPICAL EXAMPLE OF GENETIC SKIN FRAGILITY DISORDERS. GENOTYPIC HETEROGENEITY, MODIFIER GENES, EPIGENETIC, BIOCHEMICAL AND ENVIRONMENTAL FACTORS ALTER AND DETERMINE PATHOGENIC TRAITS AND, ULTIMATELY, THE WIDE AND STRIKING PHENOTYPIC VARIABILITY IN EB. BESIDES THE PRIMARY STRUCTURAL-FUNCTIONAL DEFECT, CHRONIC TISSUE DAMAGE WITH INDUCTION AND DYSREGULATION OF INFLAMMATORY PATHWAYS IS A COMMON PATHOGENIC MECHANISM IN EB. IN LOCALIZED VARIANTS, THE INFLAMMATORY ABERRATIONS MAY MAINLY AFFECT THE MICROMILIEU OF LESIONAL SKIN, WHILE A SYSTEMIC INFLAMMATORY RESPONSE WAS SHOWN TO CONTRIBUTE TO THE SYSTEMIC MORBIDITY IN SEVERE EB SUBTYPES WITH EXTENSIVE CUTANEOUS INVOLVEMENT. OUR CONTINUED UNDERSTANDING OF THE PATHOPHYSIOLOGY OF EB, AS WELL AS ADVANCES IN MOLECULAR TECHNOLOGIES, HAS PAVED THE WAY FOR TRANSLATIONAL THERAPEUTIC APPROACHES. THE SPECTRUM COMPRISES OF CORRECTIVE AND SYMPTOM-RELIEVING THERAPIES THAT INCLUDE INNOVATIVE THERAPEUTIC OPTIONS GARNERED FROM THE BENCH, REPURPOSED DRUGS APPROVED FOR OTHER DISEASES, AS WELL AS STRATEGIES FOR GENE-, PROTEIN- AND CELL-BASED THERAPIES. IMMUNOLOGICAL TRAITS FURTHER DEFINE NEW TARGETS OF THERAPY, AIMED AT IMPROVING SKIN BARRIER RESTORATION, MICROBIAL SURVEILLANCE AND INFECTION CONTROL, WOUND HEALING AND ANTI-NEOPLASTIC EFFECTS. CLINICAL AVAILABILITY AND FEASIBILITY OF THESE APPROACHES FOR ALL EB PATIENTS AND SUBTYPES ARE CURRENTLY LIMITED, REFLECTING ISSUES OF EFFICACY, SPECIFICITY, TOLERABILITY AND SAFETY. A MULTISTEP TARGETING APPROACH AND HIGHLY INDIVIDUALIZED, RISK-STRATIFIED COMBINATORY TREATMENT PLANS WILL THUS BE ESSENTIAL FOR SUSTAINED EFFICACY AND IMPROVED OVERALL QUALITY OF LIFE IN EB.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
3   703  28 BTLA EXPRESSION IN CLL: EPIGENETIC REGULATION AND IMPACT ON CLL B CELL PROLIFERATION AND ABILITY TO IL-4 PRODUCTION. IN OUR PREVIOUS STUDY, WHILE CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) CASES SHOWED HIGHER LEVELS OF B AND T LYMPHOCYTE ATTENUATOR (BTLA) MRNA COMPARED TO CONTROLS, LOWER BTLA PROTEIN EXPRESSION WAS OBSERVED IN CASES COMPARED TO CONTROLS. HENCE WE HYPOTHESIZE THAT MICRO RNA (MIR) 155-5P REGULATES BTLA EXPRESSION IN CLL. IN LINE WITH EARLIER DATA, EXPRESSION OF BTLA MRNA AND MIR-155-5P IS ELEVATED IN CLL (P = 0.034 AND P = 0.0006, RESPECTIVELY) AS WELL AS IN MEC-1 CELL LINE (P = 0.009 AND 0.016, RESPECTIVELY). INHIBITION OF MIR-155-5P PARTIALLY RESTORED BTLA PROTEIN EXPRESSION IN CLL PATIENTS (P = 0.01) AND IN MEC-1 CELL LINES (P = 0.058). ADDITIONALLY, WE AIMED TO EVALUATE THE SIGNIFICANCE OF BTLA DEFICIENCY IN CLL CELLS ON PROLIFERATION AND IL-4 PRODUCTION OF B CELLS. WE FOUND THAT SECRETION OF IL-4 IS NOT DEPENDENT ON BTLA EXPRESSION, SINCE FRACTIONS OF BTLA POSITIVE AND BTLA NEGATIVE B CELLS EXPRESSING INTRACELLULAR IL-4 WERE SIMILAR IN CLL PATIENTS AND CONTROLS. WE DEMONSTRATED THAT IN CONTROLS THE FRACTION OF PROLIFERATING CELLS IS LOWER IN BTLA POSITIVE THAN IN BTLA NEGATIVE B CELLS (P = 0.059), WHICH WAS NOT OBSERVED IN CLL. HOWEVER, THE FREQUENCY OF BTLA POSITIVE KI67+ B CELLS IN CLL WAS HIGHER COMPARED TO CORRESPONDING CELLS FROM CONTROLS (P = 0.055) WHILE THERE WERE NO DIFFERENCES BETWEEN THE EXAMINED GROUPS REGARDING FREQUENCY OF BTLA NEGATIVE KI67+ B CELLS. OUR STUDIES SUGGEST THAT MIR-155-5P IS INVOLVED IN BTLA DEFICIENCY, AFFECTING PROLIFERATION OF CLL B CELLS, WHICH MAY BE ONE OF THE MECHANISMS RESPONSIBLE FOR CLL PATHOGENESIS.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
4   969  23 CHRONIC NON-BACTERIAL OSTEOMYELITIS IS ASSOCIATED WITH IMPAIRED SP1 SIGNALING, REDUCED IL10 PROMOTER PHOSPHORYLATION, AND REDUCED MYELOID IL-10 EXPRESSION. CHRONIC NON-BACTERIAL OSTEOMYELITIS (CNO) IS AN AUTO-INFLAMMATORY DISORDER THAT AFFECTS THE SKELETAL SYSTEM. INTERLEUKIN (IL-)10 IS AN IMMUNE-MODULATORY CYTOKINE THAT CONTROLS INFLAMMATION, AND LIMITS INFLAMMATORY CYTOKINE RESPONSES. DYSREGULATION OF IL-10 EXPRESSION HAS BEEN SHOWN TO RESULT IN AUTOIMMUNE AND INFECTIOUS DISEASES. WE INVESTIGATED IL-10 EXPRESSION BY MONOCYTIC CELLS FROM CNO PATIENTS AND CONTROLS. IN RESPONSE TO STIMULATION WITH LPS, IL-10 EXPRESSION FROM CNO MONOCYTES WAS REDUCED (P<0.001). THIS WAS INDEPENDENT OF IL10 PROMOTER POLYMORPHISMS. THUS, WE INVESTIGATED SP1 RECRUITMENT TO THE IL10 PROMOTER AND SAW MARKEDLY REDUCED BINDING IN CNO MONOCYTES. THIS WAS ACCOMPANIED WITH REDUCED PHOSPHORYLATION OF HISTONE H3 SERINE 10 (H3S10), AN ACTIVATING EPIGENETIC MARK. IMPAIRED RECRUITMENT OF SP1 TO THE IL10 PROMOTER, AND REDUCED H3S10 PHOSPHORYLATION, MAY BE A REFLECTION OF DEFICIENT MAPK SIGNALING IN CNO MONOCYTES IN RESPONSE TO LPS STIMULATION. THUS, WE HAVE DISCOVERED A MECHANISM THAT MAY BE CENTRAL IN THE PATHOPHYSIOLOGY OF CNO.	2011	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
5  3636  31 INCREASED DNA METHYLTRANSFERASE ACTIVITY AND DNA METHYLATION FOLLOWING EPIDERMAL GROWTH FACTOR STIMULATION IN OVARIAN CANCER CELLS. OVARIAN CANCER PROGRESSION IS CORRELATED WITH ACCUMULATION OF ABERRANT CPG ISLAND METHYLATION. IN OVARIAN CANCER, ASCITES FLUID CONTAINS NUMEROUS EPIDERMAL-GROWTH-FACTOR-RECEPTOR (EGFR) ACTIVATORS, WHICH COULD RESULT IN A TUMOR MICROENVIRONMENT OF CONSTANT EGFR ACTIVATION. SIGNALING PATHWAYS DOWNSTREAM OF EGFR, SUCH AS RAS, REGULATE DNA METHYLATION. WE HYPOTHESIZED THAT CHRONIC EGFR ACTIVATION COULD ALTER DNA METHYLATION. WE FOUND THAT EGFR ACTIVATION INCREASED DNA METHYLTRANSFERASE (DNMT) ACTIVITY ACUTELY, AS WELL AS AFTER LONG-TERM EGF TREATMENT OR EXPRESSION OF A MUTATIONALLY ACTIVATED EGFR. FURTHERMORE, THIS INCREASE IN DNMT ACTIVITY WAS DEPENDENT ON EGFR CATALYTIC ACTIVITY AND RESULTED IN INCREASED GLOBAL DNA METHYLATION. ADDITIONALLY, TREATMENT WITH THE DNMT INHIBITOR/HYPOMETHYLATING AGENT 5-AZA-2'-DEOXYCYTIDINE (AZA) INHIBITED THE EGF INDUCED INCREASE OF BOTH DNMT ACTIVITY AND GLOBAL METHYLATION. THESE DATA SUPPORT A ROLE FOR EGFR IN THE PROCESS OF ACCUMULATED DNA METHYLATION DURING OVARIAN CANCER PROGRESSION AND SUGGEST THAT EPIGENETIC THERAPY MAY BE BENEFICIAL FOR THE TREATMENT OF OVARIAN CANCER.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
6   574  36 BCR/ABL INCREASES EZH2 LEVELS WHICH REGULATES XIAP EXPRESSION VIA MIRNA-219 IN CHRONIC MYELOID LEUKEMIA CELLS. IN THIS STUDY, WE SHOWED THAT THE LEVELS OF EZH2 IN BONE MARROW MONONUCLEAR CELLS (BMMNCS) ISOLATED FROM INDIVIDUALS WITH CHRONIC MYELOID LEUKEMIA (CML) (N=12) WERE SIGNIFICANTLY GREATER THAN THOSE IN BMMNCS ISOLATED FROM HEALTHY VOLUNTEERS (N=6) AS WELL AS INDIVIDUALS WITH PHILADELPHIA CHROMOSOME-NEGATIVE MYELOPROLIFERATIVE NEOPLASMS. LENTIVIRAL TRANSDUCTION OF THE BCR/ABL GENE IN BA/F3 CELLS INCREASED EZH2 LEVELS IN PARALLEL WITH PHOSPHORYLATION OF STAT5. NOTABLY, CHROMATIN IMMUNOPRECIPITATION ASSAYS SHOWED THAT STAT5A BOUND TO A PROMOTER REGION OF THE EZH2 GENE, RESULTING IN AN INCREASE IN THE TRANSCRIPTIONAL ACTIVITY OF EZH2 IN LEUKEMIA CELLS. IMPORTANTLY, DOWNREGULATION OF EZH2 BY SHORT HAIRPIN RNAS (SHRNAS) INHIBITED THE EXPRESSION OF XIAP AND INCREASED THE MIR-219 LEVELS ASSOCIATED WITH A DECREASE IN HYPERMETHYLATION OF MIR-219-1 CPG ISLANDS. MOREOVER, OVEREXPRESSION OF MIR-219 DECREASED THE LEVELS OF XIAP IN CML CELLS. SINCE THE 3'-UNTRANSLATED REGION (3'-UTR) OF XIAP CONTAINS MIR219-5P-COMPLEMENTARY BINDING SITE, MIR-219 MIGHT MODULATE THE EXPRESSION OF XIAP THROUGH BINDING OF MIR-219 ON THE 3'-UTR OF XIAP. TAKEN TOGETHER, BCR/ABL POSITIVELY REGULATES THE EXPRESSION OF EZH2 VIA STAT5 SIGNALING. EZH2 MODULATES EPIGENETIC CHANGES AT DNA METHYLATED REGIONS ENCODING MIR-219 AND DOWNREGULATES THE LEVEL OF MIR-219, RESULTING IN UPREGULATION OF XIAP.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
7  1061  38 CLINICAL REMISSION OF SIGHT-THREATENING NON-INFECTIOUS UVEITIS IS CHARACTERIZED BY AN UPREGULATION OF PERIPHERAL T-REGULATORY CELL POLARIZED TOWARDS T-BET AND TIGIT. BACKGROUND: NON-INFECTIOUS UVEITIS CAN CAUSE CHRONIC RELAPSING AND REMITTING OCULAR INFLAMMATION, WHICH MAY REQUIRE HIGH DOSE SYSTEMIC IMMUNOSUPPRESSION TO PREVENT SEVERE SIGHT LOSS. IT HAS BEEN CLASSICALLY DESCRIBED AS AN AUTOIMMUNE DISEASE, MEDIATED BY PRO-INFLAMMATORY TH1 AND TH17 T-CELL SUBSETS. STUDIES SUGGEST THAT NATURAL IMMUNOSUPPRESSIVE CD4(+)CD25(+)FOXP3(+) T-REGULATORY CELLS (TREGS) ARE INVOLVED IN RESOLUTION OF INFLAMMATION AND MAY BE INVOLVED IN THE MAINTENANCE OF CLINICAL REMISSION. OBJECTIVE: TO INVESTIGATE WHETHER THERE IS A PERIPHERAL BLOOD IMMUNOREGULATORY PHENOTYPE ASSOCIATED WITH CLINICAL REMISSION OF SIGHT-THREATENING NON-INFECTIOUS UVEITIS BY COMPARING PERIPHERAL BLOOD LEVELS OF TREG, TH1, AND TH17, AND ASSOCIATED DNA METHYLATION AND CYTOKINE LEVELS IN PATIENTS WITH ACTIVE UVEITIC DISEASE, CONTROL SUBJECTS AND PATIENTS (WITH PREVIOUSLY ACTIVE DISEASE) IN CLINICAL REMISSION INDUCED BY IMMUNOSUPPRESSIVE DRUGS. METHODS: ISOLATED PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMC) FROM PERIPHERAL BLOOD SAMPLES FROM PROSPECTIVELY RECRUITED SUBJECTS WERE ANALYZED BY FLOW CYTOMETRY FOR CD3, CD4, FOXP3, TIGIT, T-BET, AND RELATED ORPHAN RECEPTOR GAMMAT. EPIGENETIC DNA METHYLATION LEVELS OF FOXP3 TREG-SPECIFIC DEMETHYLATED REGION (TSDR), FOXP3 PROMOTER, TBX21, RORC2, AND TIGIT LOCI WERE DETERMINED IN CRYOPRESERVED PBMC USING A NEXT-GENERATION SEQUENCING APPROACH. RELATED CYTOKINES WERE MEASURED IN BLOOD SERA. FUNCTIONAL SUPPRESSIVE CAPACITY OF TREG WAS ASSESSED USING T-CELL PROLIFERATION ASSAYS. RESULTS: FIFTY PATIENTS WITH UVEITIS (INTERMEDIATE, POSTERIOR, AND PANUVEITIS) AND 10 CONTROL SUBJECTS WERE RECRUITED. THE FREQUENCY OF CD4(+)CD25(+)FOXP3(+) TREG, TIGIT(+) TREG, AND T-BET(+) TREG AND THE RATIO OF TREG TO TH1 WERE SIGNIFICANTLY HIGHER IN REMISSION PATIENTS COMPARED WITH PATIENTS WITH ACTIVE UVEITIC DISEASE; AND TIGIT(+) TREGS WERE A SIGNIFICANT PREDICTOR OF CLINICAL REMISSION. TREG FROM PATIENTS IN CLINICAL REMISSION DEMONSTRATED A HIGH LEVEL OF IN VITRO SUPPRESSIVE FUNCTION COMPARED WITH TREG FROM CONTROL SUBJECTS AND FROM PATIENTS WITH UNTREATED ACTIVE DISEASE. PBMC FROM PATIENTS IN CLINICAL REMISSION HAD SIGNIFICANTLY LOWER METHYLATION LEVELS AT THE FOXP3 TSDR, FOXP3 PROMOTER, AND TIGIT LOCI AND HIGHER LEVELS AT RORC LOCI THAN THOSE WITH ACTIVE DISEASE. CLINICAL REMISSION WAS ALSO ASSOCIATED WITH SIGNIFICANTLY HIGHER SERUM LEVELS OF TRANSFORMING GROWTH FACTOR BETA AND IL-10, WHICH POSITIVELY CORRELATED WITH TREG LEVELS, AND LOWER SERUM LEVELS OF IFNGAMMA, IL-17A, AND IL-22 COMPARED WITH PATIENTS WITH ACTIVE DISEASE. CONCLUSION: CLINICAL REMISSION OF SIGHT-THREATENING NON-INFECTIOUS UVEITIS HAS AN IMMUNOREGULATORY PHENOTYPE CHARACTERIZED BY UPREGULATION OF PERIPHERAL TREG, POLARIZED TOWARD T-BET AND TIGIT. THESE FINDINGS MAY ASSIST WITH INDIVIDUALIZED THERAPY OF UVEITIS, BY INFORMING WHETHER DRUG THERAPY HAS INDUCED PHENOTYPICALLY STABLE TREG ASSOCIATED WITH LONG-TERM CLINICAL REMISSION.	2018	

8  5399  29 REDUCED TEN-ELEVEN TRANSLOCATION AND ISOCITRATE DEHYDROGENASE EXPRESSION IN INFLAMMATORY HIDRADENITIS SUPPURATIVA LESIONS. BACKGROUND: AS ENVIRONMENTAL FACTORS APPEAR TO PREDISPOSE PATIENTS TO HIDRADENITIS SUPPURATIVA (HS), STUDYING EPIGENETIC MODIFICATIONS IS OF INTEREST TO FURTHER UNDERSTAND THE PATHOGENESIS OF HS. OBJECTIVES: TO STUDY THE EXPRESSION OF DNA HYDROXYMETHYLATION REGULATORS, NAMELY THE TEN-ELEVEN TRANSLOCATION (TET) AND ISOCITRATE DEHYDROGENASE (IDH) FAMILY, IN THE SKIN OF HS PATIENTS. MATERIALS & METHODS: TWENTY PATIENTS WITH HS AND 12 HEALTHY SUBJECTS WERE RECRUITED. WE ANALYSED THE EXPRESSION OF TET1, TET2, TET3, IDH1, IDH2, IDH3A, AND IDH3B IN LESIONAL AND PERILESIONAL HS TISSUE AS WELL AS TISSUE FROM HEALTHY CONTROLS BY QUANTITATIVE REAL-TIME REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION (RT-PCR). IN ADDITION, IMMUNOHISTOCHEMISTRY WAS PERFORMED FOR TET1, TET2, AND TET3. RESULTS: RT-PCR ANALYSIS SHOWED THAT MRNA OF ALL THE STUDIED GENES WAS SIGNIFICANTLY UNDER-EXPRESSED IN LESIONAL HS SKIN COMPARED TO HEALTHY SKIN. IDH1 AND IDH2 MRNA EXPRESSION WAS ALSO SIGNIFICANTLY LOWER IN PERILESIONAL HS SKIN COMPARED TO HEALTHY SKIN, AND TET3 MRNA EXPRESSION WAS SIGNIFICANTLY LOWER IN LESIONAL HS SKIN COMPARED TO PERILESIONAL HS SKIN. RT-PCR ANALYSIS FOR TET1, TET2, AND TET3 MRNA EXPRESSION WAS CONFIRMED BY IMMUNOHISTOCHEMICAL ANALYSIS. CORRELATION ANALYSIS REVEALED A SIGNIFICANT POSITIVE CORRELATION BETWEEN TET AND IDH GENE EXPRESSION IN PERILESIONAL AND LESIONAL HS SKIN. CONCLUSIONS: OUR RESULTS SUGGEST THAT EPIGENETIC CHANGES OCCUR IN HS TISSUE AND THAT ABERRANT EXPRESSION OF THE DNA HYDROXYMETHYLATION REGULATORS MAY PLAY A ROLE IN THE PATHOGENESIS OF HS. AS EPIGENETIC MODIFICATIONS ARE REVERSIBLE, FURTHER RESEARCH INTO THE CAUSE OF THESE ABERRANT EXPRESSION PATTERNS IS WARRANTED IN ORDER TO DEVELOP POSSIBLE NOVEL THERAPEUTIC APPROACHES.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
9  6336  42 THE ROLE OF DNA METHYLATION IN TRANSCRIPTIONAL REGULATION OF PRO-NOCICEPTIVE GENES IN RAT TRIGEMINAL GANGLIA. EPIGENETIC MODULATION BY DNA METHYLATION IS ASSOCIATED WITH ABERRANT GENE EXPRESSION IN SENSORY NEURONS, WHICH CONSEQUENTLY LEADS TO PATHOLOGICAL PAIN RESPONSES. IN THIS STUDY, WE SOUGHT TO INVESTIGATE WHETHER PERIPHERAL INFLAMMATION ALTERS GLOBAL DNA METHYLATION IN TRIGEMINAL GANGLIA (TG) AND RESULTS IN ABNORMAL EXPRESSION OF PRO-NOCICEPTIVE GENES. OUR RESULTS SHOW THAT PERIPHERAL INFLAMMATION REMOTELY REDUCED THE LEVEL OF GLOBAL DNA METHYLATION IN RAT TG WITH A CONCURRENT REDUCTION IN DNMT1 AND DNMT3A EXPRESSION. USING UNBIASED STEPS, WE SELECTED THE FOLLOWING PRO-NOCICEPTIVE CANDIDATE GENES THAT ARE POTENTIALLY REGULATED BY DNA METHYLATION: TRPV1, TRPA1, P2X3, AND PIEZO2. INHIBITION OF DNMT WITH 5-AZA-DC IN DISSOCIATED TG CELLS PRODUCED DOSE-DEPENDENT UPREGULATION OF TRPV1, TRPA1, AND P2X3. SYSTEMIC TREATMENT OF ANIMALS WITH 5-AZA-DC SIGNIFICANTLY INCREASED THE EXPRESSION OF TRPV1, TRPA1, AND PIEZO2 IN TG. FURTHERMORE, THE OVEREXPRESSION OF DNMT3A, AS DELIVERED BY A LENTIVIRAL VECTOR, SIGNIFICANTLY DOWNREGULATED TRPV1 AND PIEZO2 EXPRESSION AND ALSO RELIABLY DECREASED TRPA1 AND P2X3 TRANSCRIPTS. MEDIP REVEALED THAT THIS OVEREXPRESSION ALSO SIGNIFICANTLY ENHANCED METHYLATION OF CGIS ASSOCIATED WITH TRPV1 AND TRPA1. IN ADDITION, BISULFITE SEQUENCING DATA INDICATED THAT THE CGI ASSOCIATED WITH TRPA1 WAS METHYLATED IN A PATTERN CATALYZED BY DNMT3A. TAKEN TOGETHER, OUR RESULTS SHOW THAT ALL 4 PRO-NOCICEPTIVE GENES ARE SUBJECT TO EPIGENETIC MODULATION VIA DNA METHYLATION, LIKELY VIA DNMT3A UNDER INFLAMMATORY CONDITIONS. THESE FINDINGS PROVIDE THE FIRST EVIDENCE FOR THE FUNCTIONAL IMPORTANCE OF DNA METHYLATION AS AN EPIGENETIC FACTOR IN THE TRANSCRIPTION OF PRO-NOCICEPTIVE GENES IN TG THAT ARE IMPLICATED IN PATHOLOGICAL OROFACIAL PAIN RESPONSES.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
10  3953  33 LOCUS-SPECIFIC REVERSIBLE DNA METHYLATION REGULATES TRANSIENT IL-10 EXPRESSION IN TH1 CELLS. IL-10 IS A PLEIOTROPIC CYTOKINE WITH MULTIFACETED FUNCTIONS IN ESTABLISHING IMMUNE HOMEOSTASIS. ALTHOUGH EXPRESSED BY TH1 AND TH2 CELLS, CONVENTIONAL TH1 CELLS PRODUCE MARGINAL LEVELS OF IL-10 COMPARED WITH THEIR TH2 COUNTERPARTS. IN THIS STUDY, WE INVESTIGATED THE EPIGENETIC MECHANISMS OF IL-10 GENE EXPRESSION IN TH1 CELLS. BIOINFORMATICS EMBOSS CPG PLOT ANALYSIS AND BISULFITE PYROSEQUENCING REVEALED THREE CPG DNA METHYLATION SITES IN THE IL-10 GENE LOCUS. PROGRESSIVE DNA METHYLATION AT ALL OF THE CPG REGIONS OF INTEREST (ROIS) ESTABLISHED A REPRESSIVE PROGRAM OF IL-10 GENE EXPRESSION IN TH1 CELLS. INTERESTINGLY, TH1 CELLS TREATED WITH IL-12 AND IL-27 CYTOKINES, THEREBY MIMICKING A CHRONIC INFLAMMATORY CONDITION IN VIVO, DISPLAYED A SIGNIFICANT INCREASE IN IL-10 PRODUCTION THAT WAS ACCOMPANIED BY SELECTIVE DNA DEMETHYLATION AT ROI 3 LOCATED IN INTRON 3. IL-10-PRODUCING T CELLS ISOLATED FROM LYMPHOCYTIC CHORIOMENINGITIS VIRUS-INFECTED MICE ALSO SHOWED ENHANCED DNA DEMETHYLATION AT ROI 3. BINDING OF STAT1 AND STAT3 TO DEMETHYLATED ROI 3 ENHANCED IL-10 EXPRESSION IN AN IL-12/IL-27-DEPENDENT MANNER. ACCORDINGLY, CD4(+) T CELLS ISOLATED FROM STAT1- OR STAT3-KNOCKOUT MICE WERE SIGNIFICANTLY DEFECTIVE IN IL-10 PRODUCTION. OUR DATA SUGGEST THAT, ALTHOUGH STABLY MAINTAINED DNA METHYLATION AT THE PROMOTER MAY REPRESS IL-10 EXPRESSION IN TH1 CELLS, LOCUS-SPECIFIC REVERSIBLE DNA DEMETHYLATION MAY SERVE AS A THRESHOLD PLATFORM TO CONTROL TRANSIENT IL-10 GENE EXPRESSION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
11  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	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
12  3645  28 INCREASED PRESENCE AND DIFFERENTIAL MOLECULAR IMPRINTING OF TRANSIT AMPLIFYING CELLS IN PSORIASIS. PSORIASIS IS A VERY COMMON CHRONIC INFLAMMATORY SKIN DISEASE CHARACTERIZED BY EPIDERMAL THICKENING AND SCALING RESULTING FROM KERATINOCYTE HYPERPROLIFERATION AND IMPAIRED DIFFERENTIATION. PATHOMECHANISTIC STUDIES IN PSORIASIS ARE OFTEN LIMITED BY USING WHOLE SKIN TISSUE BIOPSIES, NEGLECTING THEIR STRATIFICATION AND CELLULAR DIVERSITY. THIS STUDY AIMED AT CHARACTERIZING EPIDERMAL ALTERATIONS IN PSORIASIS AT THE LEVEL OF KERATINOCYTE POPULATIONS. EPIDERMAL CELL POPULATIONS WERE PURIFIED FROM SKIN BIOPSIES OF PSORIASIS PATIENTS AND HEALTHY DONORS USING A NOVEL CELL TYPE-SPECIFIC APPROACH. MOLECULAR CHARACTERIZATION OF THE TRANSIT-AMPLIFYING CELLS (TAC), THE KEY PLAYERS OF EPIDERMAL RENEWAL, WAS PERFORMED USING IMMUNOCYTOFLUORESCENCE-TECHNIQUE AND INTEGRATED MULTISCALE-OMICS ANALYSES. ALREADY TAC FROM NON-LESIONAL PSORIATIC SKIN SHOWED ALTERED METHYLATION AND DIFFERENTIAL EXPRESSION IN 1.7% AND 1.0% OF ALL PROTEIN-CODING GENES, RESPECTIVELY. IN PSORIATIC LESIONS, TAC WERE STRONGLY EXPANDED SHOWING FURTHER INCREASED DIFFERENTIALLY METHYLATED (10-FOLD) AND EXPRESSED (22-FOLD) GENES NUMBERS. IMPORTANTLY, 17.2% OF DIFFERENTIALLY EXPRESSED GENES WERE ASSOCIATED WITH RESPECTIVE GENE METHYLATIONS. COMPARED WITH NON-LESIONAL TAC, PATHWAY ANALYSES REVEALED METABOLIC ALTERATIONS AS ONE FEATURE PREDOMINANTLY CHANGED IN TAC DERIVED FROM ACTIVE PSORIATIC LESIONS. OVERALL, OUR STUDY SHOWED STAGE-SPECIFIC MOLECULAR ALTERATIONS, ALLOWS NEW INSIGHTS INTO THE PATHOGENESIS, AND IMPLIES THE INVOLVEMENT OF EPIGENETIC MECHANISMS IN LESION DEVELOPMENT IN PSORIASIS. KEY MESSAGES: TRANSIT AMPLIFYING CELL (TAC) NUMBERS ARE HIGHLY INCREASED IN PSORIATIC LESIONS PSORIATIC TAC SHOW PROFOUND MOLECULAR ALTERATIONS & STAGE-SPECIFIC IDENTITY TAC FROM UNAFFECTED AREAS ALREADY SHOW FIRST SIGNS OF MOLECULAR ALTERATIONS LESIONAL TAC SHOW A PREFERENCE IN METABOLIC-RELATED ALTERATIONS.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
13  5049  30 PHARMACOLOGICAL INHIBITION OF RORC2 ENHANCES HUMAN TH17-TREG STABILITY AND FUNCTION. INFLAMMATORY BOWEL DISEASES (IBD) ARE CHRONIC CONDITIONS THAT RESULT FROM UNCONTROLLED INTESTINAL INFLAMMATION. PATHOGENIC TH17 CELLS, CHARACTERIZED BY PRODUCTION OF IL-17A IN THE ABSENCE OF IL-10, ARE THOUGHT TO CONTRIBUTE TO THIS INFLAMMATION, BUT IN HUMANS, ANTIBODY-MEDIATED BLOCKADE OF IL-17A IS AN INEFFECTIVE IBD THERAPY WHEREAS IL-23 BLOCKADE IS EFFECTIVE. HERE, WE INVESTIGATED THE EFFECTS OF PHARMACOLOGICAL INHIBITION OF RORC2, THE TH17 CELL LINEAGE-DEFINING TRANSCRIPTION FACTOR, ON IN VIVO-DIFFERENTIATED HUMAN TH17 CELLS AND TH17-LIKE TREGS (TH17-TREGS). BMS-336, A SMALL MOLECULE RORC2 INVERSE AGONIST, INHIBITED EXPRESSION OF RORC2-REGULATED GENES IN PERIPHERAL TH17 CELLS (CD4(+) CD25(-) CD127(+) CXCR3(-) CCR4(+) CCR6(+) ) IN A DOSE-DEPENDENT MANNER, WITH SIMILAR INHIBITORY EFFECTS ON LAMINAR PROPRIA MONONUCLEAR CELLS FROM IBD AND NON-IBD SUBJECTS. EXPOSURE OF PERIPHERAL TH17-TREGS (CD4(+) CD25(HI) CD127(LO) CXCR3(-) CCR4(+) CCR6(+) ) TO BMS-336 ALSO INHIBITED IL-17A PRODUCTION AND PREVENTED INFLAMMATORY CYTOKINE-INDUCED DESTABILIZATION, AS EVIDENCED BY PRESERVED FOXP3 EXPRESSION AND EPIGENETIC STATUS OF THE TREG-SPECIFIC DEMETHYLATION REGION. IN PARALLEL, RORC2 INHIBITION INCREASED THE PRODUCTION OF IL-10 IN TH17-TREGS, RESULTING IN ENHANCED SUPPRESSION OF INFLAMMATORY CYTOKINES FROM MYELOID CELLS. THUS, VIA ITS ABILITY TO SIMULTANEOUSLY INHIBIT TH17 CELLS AND ENHANCE THE STABILITY AND FUNCTION OF TH17-TREGS, PHARMACOLOGICAL INHIBITION OF RORC2 IS A PROMISING APPROACH TO SUPPRESS INFLAMMATION AND PROMOTE IMMUNE REGULATION IN IBD.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
14  2760  30 EXPRESSION OF PD-L1, PD-L2, PD-1 AND CTLA4 IN MYELODYSPLASTIC SYNDROMES IS ENHANCED BY TREATMENT WITH HYPOMETHYLATING AGENTS. BLOCKADE OF IMMUNE CHECKPOINTS IS EMERGING AS A NEW FORM OF ANTICANCER THERAPY. WE STUDIED THE EXPRESSION OF PROGRAMMED DEATH LIGAND 1 (PD-L1), PD-L2, PROGRAMMED DEATH 1 (PD-1) AND CYTOTOXIC T LYMPHOCYTE-ASSOCIATED ANTIGEN 4 (CTLA4) MRNA IN CD34+ CELLS FROM MYELODYSPLASTIC SYNDROME (MDS), CHRONIC MYELOMONOCYTIC LEUKEMIA (CMML) AND ACUTE MYELOID LEUKEMIA (AML) PATIENTS (N=124). ABERRANT UPREGULATION (?2-FOLD) WAS OBSERVED IN 34, 14, 15 AND 8% OF THE PATIENTS. INCREASED EXPRESSION OF THESE FOUR GENES WAS ALSO OBSERVED IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMNCS) (N=61). THE RELATIVE EXPRESSION OF PD-L1 FROM PBMNC WAS SIGNIFICANTLY HIGHER IN MDS (P=0.018) AND CMML (P=0.0128) COMPARED WITH AML. BY IMMUNOHISTOCHEMICAL ANALYSIS, PD-L1 PROTEIN EXPRESSION WAS OBSERVED IN MDS CD34+ CELLS, WHEREAS STROMA/NON-BLAST CELLULAR COMPARTMENT WAS POSITIVE FOR PD-1. IN A COHORT OF PATIENTS TREATED WITH EPIGENETIC THERAPY, PD-L1, PD-L2, PD-1 AND CTLA4 EXPRESSION WAS UPREGULATED. PATIENTS RESISTANT TO THERAPY HAD RELATIVE HIGHER INCREMENTS IN GENE EXPRESSION COMPARED WITH PATIENTS WHO ACHIEVED RESPONSE. TREATMENT OF LEUKEMIA CELLS WITH DECITABINE RESULTED IN A DOSE-DEPENDENT UPREGULATION OF ABOVE GENES. EXPOSURE TO DECITABINE RESULTED IN PARTIAL DEMETHYLATION OF PD-1 IN LEUKEMIA CELL LINES AND HUMAN SAMPLES. THIS STUDY SUGGESTS THAT PD-1 SIGNALING MAY BE INVOLVED IN MDS PATHOGENESIS AND RESISTANCE MECHANISMS TO HYPOMETHYLATING AGENTS. BLOCKADE OF THIS PATHWAY CAN BE A POTENTIAL THERAPY IN MDS AND AML.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
15  5459  34 RESEARCH ON THE EPIGENETIC REGULATION MECHANISM OF THE PTPN6 GENE IN ADVANCED CHRONIC MYELOID LEUKAEMIA. PTPN6, A TYROSINE PHOSPHATASE PROTEIN, PLAYS A NEGATIVE ROLE IN CELL SIGNAL TRANSDUCTION AND IS NEGATIVELY CORRELATED WITH TUMOUR FORMATION AND GROWTH. HOWEVER, EPIGENETIC REGULATION MECHANISM OF THE PTPN6 GENE IN ADVANCED CHRONIC MYELOID LEUKAEMIA (CML) REMAINS UNCLEAR. THIS STUDY INVESTIGATED BONE MARROW OR BLOOD SAMPLES FROM 44 CML PATIENTS AND 10 HEALTHY VOLUNTEERS. KCL22 AND K562 CELLS WERE CULTURED AND TREATED WITH DEMETHYLATION DRUGS AND HISTONE DEACETYLASE INHIBITORS. REAL TIME QUANTITATIVE POLYMERASE CHAIN REACTION (QPCR), METHYLATION-SPECIFIC PCR, BISULFITE SEQUENCING PCR, WESTERN BLOTTING, CO-IMMUNOPRECIPITATION AND CHROMATIN IMMUNOPRECIPITATION (CHIP) WAS PERFORMED. PTPN6 WAS DOWN-REGULATED IN CELL LINES AND PATIENTS WITH ADVANCED PHASE CML, WHEREAS DNMT1, DNMT3A, MECP2, MBD2 AND HDAC1 WERE UP-REGULATED. TREATMENT WITH 5-AZACYTIDINE, DECITABINE, SODIUM VALPROATE AND LBH589 INCREASED PTPN6 EXPRESSION, BUT DECREASED THAT OF DNMT1, DNMT3A, MECP2, MBD2 AND HDAC1. IMMUNOPRECIPITATION AND MASS SPECTROMETRY SHOWED THAT HDAC1 COMBINED DIRECTLY WITH PTPN6. CHIP-SEQ SHOWED THAT HDAC1 DID NOT COMBINE WITH THE PROMOTER REGION OF PTPN6, WHILE MAPK, AKT, STAT5, JAK2 AND MYC PROMOTER REGIONS ALL COMBINED WITH HDAC1. PTPN6 IS ASSOCIATED WITH PROGRESSION OF CML. LOW EXPRESSION LEVEL OF PTPN6 WAS ASSOCIATED WITH DNA METHYLATION AND REGULATED BY HISTONE ACETYLATION. HDAC1 PARTICIPATES IN THE REGULATION OF PTPN6.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
16  1297  38 DECREASED MRNA EXPRESSION LEVELS OF DNA METHYLTRANSFERASES TYPE 1 AND 3A IN SYSTEMIC LUPUS ERYTHEMATOSUS. OBJECTIVES: SYSTEMIC LUPUS ERYTHEMATOSUS (SLE) IS A CHRONIC RELAPSING AUTOIMMUNE DISEASE CHARACTERIZED BY THE PRESENCE OF AUTOANTIBODIES DIRECTED AGAINST NUCLEAR ANTIGENS AND BY CHRONIC INFLAMMATION. ALTHOUGH THE ETIOLOGY OF SLE REMAINS UNCLEAR, THE INFLUENCE OF ENVIRONMENT FACTORS, WHICH IS LARGELY REFLECTED BY THE EPIGENETIC MECHANISMS, WITH DNA METHYLATION CHANGES IN PARTICULAR, IS GENERALLY CONSIDERED AS MAIN PLAYERS IN THE PATHOGENESIS OF SLE. WE STUDIED DNA METHYLTRANSFERASES' (DNMTS) TYPE 1, 3A AND 3B TRANSCRIPT LEVELS IN PERIPHERAL BLOOD MONONUCLEAR CELLS FROM PATIENTS DIAGNOSED WITH SYSTEMIC LUPUS ERYTHEMATOSUS AND FROM THE HEALTHY CONTROL SUBJECTS. FURTHERMORE, THE ASSOCIATION OF DNMT1, DNMT3A, AND DNMT3B MRNA LEVELS WITH GENDER, AGE, AND MAJOR CLINICAL MANIFESTATIONS WAS ANALYZED. METHODS: PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) WERE ISOLATED FROM 32 SLE PATIENTS AND 40 HEALTHY CONTROLS. REVERSE TRANSCRIPTION AND REAL-TIME QUANTITATIVE POLYMERASE CHAIN REACTION (RT-QPCR) ANALYSES WERE USED TO DETERMINE DNMT1, DNMT3A, AND DNMT3B MRNA EXPRESSION LEVELS. RESULTS: SIGNIFICANTLY LOWER DNMT1 (P = 0.015543) AND DNMT3A (P = 0.003652) TRANSCRIPT LEVELS IN SLE PATIENTS WERE OBSERVED COMPARED WITH HEALTHY CONTROLS. NEVERTHELESS, THE DNMT3B MRNA EXPRESSION LEVELS WERE MARKEDLY LOWER COMPARED WITH DNMT1 AND DNMT3A, BOTH IN PBMCS FROM AFFECTED PATIENTS AND THOSE FROM CONTROL SUBJECTS. FURTHERMORE, THE DNMT1 TRANSCRIPT LEVELS WERE POSITIVELY CORRELATED WITH SLE DISEASE ACTIVITY INDEX (SLEDAI) (R (S) = 0.4087, P = 0.020224), WHILE THE DNMT3A TRANSCRIPT LEVELS WERE NEGATIVELY CORRELATED WITH PATIENTS AGE (R (S) = -0.3765, P = 0.03369). CONCLUSIONS: OUR ANALYSES CONFIRMED THE IMPORTANCE OF EPIGENETIC ALTERATIONS IN SLE ETIOLOGY. MOREOVER, OUR RESULTS SUGGEST THAT THE PRESENCE OF SOME CLINICAL MANIFESTATIONS, SUCH AS PHOTOTOSENSITIVITY AND ARTHRITIS, MIGHT BE ASSOCIATED WITH THE DYSREGULATION OF DNA METHYLTRANSFERASES' MRNA EXPRESSION LEVELS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
17  5417  47 REGULATION OF DNA METHYLATION IN RHEUMATOID ARTHRITIS SYNOVIOCYTES. RHEUMATOID ARTHRITIS (RA) IS A CHRONIC INFLAMMATORY DISEASE IN WHICH FIBROBLAST-LIKE SYNOVIOCYTES (FLS) EXHIBIT AN AGGRESSIVE PHENOTYPE. ALTHOUGH THE MECHANISMS RESPONSIBLE ARE NOT WELL DEFINED, EPIGENETIC DETERMINANTS SUCH AS DNA METHYLATION MIGHT CONTRIBUTE. DNA METHYLTRANSFERASES (DNMTS) ARE CRITICAL ENZYMES THAT ESTABLISH AND MAINTAIN DNA METHYLATION. WE EVALUATED WHETHER PROINFLAMMATORY CYTOKINES MIGHT CONTRIBUTE TO DIFFERENTIAL DNA METHYLATION PREVIOUSLY DESCRIBED IN RA FLS THROUGH ALTERED DNMT EXPRESSION. FLS WERE OBTAINED FROM RA AND OSTEOARTHRITIS (OA) SYNOVIUM AT THE TIME OF TOTAL JOINT REPLACEMENT. GENE EXPRESSION WAS DETERMINED BY QUANTITATIVE REAL-TIME PCR AND PROTEIN EXPRESSION BY WESTERN BLOT ANALYSIS. DNMT ACTIVITY WAS MEASURED WITH A FUNCTIONAL ASSAY, AND GLOBAL METHYLATION WAS DETERMINED BY AN IMMUNOASSAY THAT DETECTS METHYLCYTOSINE. RESTING EXPRESSION OF DNMT1, -3A, AND -3B MRNA WERE SIMILAR IN RA AND OA FLS. WESTERN BLOT SHOWED ABUNDANT DNMT1 AND DNMT3A PROTEIN. EXPOSURE TO IL-1 DECREASED DNMT1 AND DNMT3A MRNA EXPRESSION IN FLS. DOSE RESPONSES DEMONSTRATED DECREASED DNMT EXPRESSION AT CONCENTRATIONS AS LOW AS 1 PG/ML OF IL-1. DNMT MRNA LEVELS DECREASED RAPIDLY, WITH SIGNIFICANT SUPPRESSION AFTER 2-8 H OF IL-1 STIMULATION. IL-1 STIMULATION OF OA FLS DID NOT AFFECT METHYLATION OF LINE1 SITES BUT LED TO DEMETHYLATION OF A CHI3L1 LOCUS THAT IS HYPOMETHYLATED IN RA FLS. CHRONIC IL-1 STIMULATION ALSO MIMICKED THE EFFECT OF A DNMT INHIBITOR ON FLS GENE EXPRESSION. EXPOSURE TO PROINFLAMMATORY MEDIATORS REVERSIBLY ALTERS DNA METHYLATION IN FLS BY DECREASING DNMT EXPRESSION AND FUNCTION. THESE DATA SUGGEST THAT IL-1 CAN POTENTIALLY IMPRINT CELLS IN CHRONIC INFLAMMATORY DISEASES.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
18   150  32 ABERRANT HYPOMETHYLATION OF THE CANCER-TESTIS ANTIGEN PRAME CORRELATES WITH PRAME EXPRESSION IN ACUTE MYELOID LEUKEMIA. PRAME IS A TUMOR-ASSOCIATED ANTIGEN, WHICH BELONGS TO THE FAMILY OF CANCER-TESTIS ANTIGENS (CTA). THE EXPRESSION OF CTA IS MAINLY RESTRICTED TO THE TESTIS AND VARIOUS TUMORS. IN CONTRAST TO OTHER CTA, PRAME EXPRESSION IS ALSO FREQUENTLY DETECTED IN ACUTE AND CHRONIC LEUKEMIAS. DUE TO THIS EXPRESSION PATTERN, PRAME HAS ATTRACTED GREAT INTEREST AS A PROGNOSTIC TUMOR MARKER THAT CAN BE USED FOR THE DETECTION OF MINIMAL RESIDUAL DISEASE AND AS A POTENTIAL TARGET FOR IMMUNOTHERAPY. IN ACUTE MYELOID LEUKEMIA (AML), PRAME EXPRESSION HAS BEEN OBSERVED IN 30-64% OF CASES. TO EVALUATE WHETHER EPIGENETIC MECHANISMS CONTRIBUTE TO PRAME ACTIVATION IN AML, WE STUDIED DNA METHYLATION OF 15 CPG DINUCLEOTIDES WITHIN A CPG-RICH REGION LOCATED IN THE INTRON 1 OF THE PRAME GENE. DNA METHYLATION WAS DETERMINED BY SEQUENCE ANALYSIS OF CLONED PCR PRODUCTS GENERATED FROM BISULFITE-TREATED GENOMIC DNA. METHYLATION PATTERNS WERE CORRELATED WITH PRAME MRNA LEVELS AS DETERMINED BY MICROARRAY ANALYSIS AND REAL-TIME PCR. WE FOUND ALMOST COMPLETE METHYLATION IN MONONUCLEAR BLOOD CELLS FROM TWO HEALTHY DONORS AND IN BONE MARROW CELLS OF FOUR PRAME-NEGATIVE AML PATIENTS. IN CONTRAST, THE DEGREE OF PRAME METHYLATION WAS CLEARLY REDUCED IN FOUR PRAME-POSITIVE AML BONE MARROW SAMPLES. IN PARTICULAR, THESE SAMPLES WERE CHARACTERIZED BY THE PRESENCE OF CLONES, WHICH WERE COMPLETELY DEVOID OF METHYLATION. THE SIGNIFICANT INVERSE CORRELATION BETWEEN THE DEGREE OF METHYLATION AND PRAME EXPRESSION SUGGESTS A CAUSAL ROLE OF DNA METHYLATION IN PRAME REGULATION. SUCH A ROLE IS FURTHER SUPPORTED BY THE OBSERVATION THAT TREATMENT OF PRAME-NEGATIVE CELL LINES U-937 AND THP-1 WITH THE DEMETHYLATING AGENT 5'-AZA-2'DC RESULTED IN A DOSE-RELATED UPREGULATION OF PRAME EXPRESSION.	2008	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
19   547  23 ATTENUATED TLR4/MAPK SIGNALING IN MONOCYTES FROM PATIENTS WITH CRMO RESULTS IN IMPAIRED IL-10 EXPRESSION. CHRONIC RECURRENT MULTIFOCAL OSTEOMYELITIS (CRMO) IS AN AUTOINFLAMMATORY BONE DISORDER OF UNKNOWN ORIGIN. WE PREVIOUSLY DEMONSTRATED THAT MONOCYTES FROM CRMO PATIENTS FAIL TO EXPRESS THE IMMUNE-MODULATORY CYTOKINE INTERLEUKIN-10 (IL-10) IN A CHROMATIN DEPENDENT MANNER. HERE, WE DEMONSTRATE THAT ATTENUATED EXTRACELLULAR-SIGNAL REGULATED KINASE (ERK)1 AND 2 SIGNALING IN RESPONSE TO TLR4 ACTIVATION RESULTS IN FAILURE TO INDUCE IL-10 EXPRESSION IN MONOCYTES FROM CRMO PATIENTS. ATTENUATED ERK1/2 ACTIVATION RESULTS IN 1) REDUCED LEVELS OF SP-1, A TRANSCRIPTION FACTOR THAT INDUCES IL-10 EXPRESSION IN MONOCYTES, AND 2) IMPAIRED H3S10 PHOSPHORYLATION OF THE IL10 PROMOTER, AN ACTIVATING EPIGENETIC MARK. THE PRO-INFLAMMATORY CYTOKINES TUMOR NECROSIS FACTOR (TNF)ALPHA AND IL-6 ARE NOT NEGATIVELY AFFECTED, RESULTING IN AN IMBALANCE TOWARDS PRO-INFLAMMATORY CYTOKINES. THUS, IMPAIRED ERK1/2 SIGNALING WITH SUBSEQUENTLY REDUCED SP-1 EXPRESSION AND H3S10 PHOSPHORYLATION OF THE IL10 PROMOTER MAY CENTRALLY CONTRIBUTE TO THE PATHOPHYSIOLOGY OF CRMO.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               
20  6661  40 UPREGULATION OF DNA METHYLTRANSFERASE-MEDIATED GENE SILENCING, ANCHORAGE-INDEPENDENT GROWTH, AND MIGRATION OF COLON CANCER CELLS BY INTERLEUKIN-6. INFLAMMATORY BOWEL DISEASE IS CHARACTERIZED BY CHRONIC INFLAMMATION WHICH PREDISPOSES TO COLORECTAL CANCER. THE MECHANISMS BY WHICH INFLAMMATION PROMOTES TUMORIGENESIS ARE NOT FULLY KNOWN. WE AIMED TO INVESTIGATE THE LINKS BETWEEN COLONIC INFLAMMATION AND TUMORIGENESIS VIA EPIGENETIC GENE SILENCING. COLON CANCER SPECIMENS WERE ASSESSED FOR THE EXPRESSION OF DNA METHYLTRANSFERASE-1 (DNMT-1) USING IMMUNOHISTOCHEMISTRY. COLORECTAL CARCINOMA CELL LINES WERE ASSESSED FOR DNMT1 EXPRESSION, METHYLCYTOSINE CONTENT, PROMOTER METHYLATION, GENE EXPRESSION, AND TUMORIGENESIS IN RESPONSE TO INTERLEUKIN (IL)-6. DNMT1 WAS EXPRESSED AT HIGHER LEVELS IN BOTH THE PERITUMORAL STROMA AND TUMOR IN INFLAMMATORY BOWEL DISEASE-ASSOCIATED CANCERS COMPARED WITH SPORADIC COLON CANCERS. IL-6 TREATMENT OF COLON CANCER CELLS RESULTED IN AN INCREASE IN DNMT1 EXPRESSION, INDEPENDENT OF DE NOVO GENE EXPRESSION. IL-6 INCREASED THE METHYLATION OF PROMOTER REGIONS OF GENES ASSOCIATED WITH TUMOR SUPPRESSION, ADHESION, AND APOPTOSIS RESISTANCE. EXPRESSION OF A SUBSET OF THESE GENES WAS DOWNREGULATED BY IL-6, AN EFFECT THAT WAS PREVENTED BY PREINCUBATION WITH 5-AZADEOXYCYTIDINE, A DNMT1 INHIBITOR. ANCHORAGE-INDEPENDENT GROWTH AND MIGRATION OF COLON CANCER CELLS WAS ALSO INCREASED BY IL-6 IN A 5-AZADEOXYCYTIDINE-SENSITIVE MANNER. OUR RESULTS INDICATE THAT DNMT-MEDIATED GENE SILENCING MAY PLAY A ROLE IN INFLAMMATION-ASSOCIATED COLON TUMORIGENESIS.	2010