1 5527 175 RNA HELICASE DEAD BOX PROTEIN 5 REGULATES POLYCOMB REPRESSIVE COMPLEX 2/HOX TRANSCRIPT ANTISENSE INTERGENIC RNA FUNCTION IN HEPATITIS B VIRUS INFECTION AND HEPATOCARCINOGENESIS. CHRONIC HEPATITIS B VIRUS (HBV) INFECTION IS A MAJOR FACTOR IN HEPATOCELLULAR CARCINOMA (HCC) PATHOGENESIS BY A MECHANISM NOT YET UNDERSTOOD. ELUCIDATING MECHANISMS OF HBV-MEDIATED HEPATOCARCINOGENESIS IS NEEDED TO GAIN INSIGHTS INTO CLASSIFICATION AND TREATMENT OF HCC. IN HBV REPLICATING CELLS, INCLUDING VIRUS-ASSOCIATED HCCS, SUPPRESSOR OF ZESTE 12 HOMOLOG (SUZ12), A CORE SUBUNIT OF POLYCOMB REPRESSIVE COMPLEX2 (PRC2), UNDERGOES PROTEASOMAL DEGRADATION. THIS PROCESS REQUIRES THE LONG NONCODING RNA, HOX TRANSCRIPT ANTISENSE INTERGENIC RNA (HOTAIR). INTRIGUINGLY, HOTAIR INTERACTS WITH PRC2 AND ALSO BINDS RNA-BINDING E3 LIGASES, SERVING AS A UBIQUITINATION SCAFFOLD. HEREIN, WE IDENTIFIED THE RNA HELICASE, DEAD BOX PROTEIN 5 (DDX5), AS A REGULATOR OF SUZ12 STABILITY AND PRC2-MEDIATED GENE REPRESSION, ACTING BY REGULATING RNA-PROTEIN COMPLEXES FORMED WITH HOTAIR. SPECIFICALLY, KNOCKDOWN OF DDX5 AND/OR HOTAIR ENABLED REEXPRESSION OF PRC2-REPRESSED GENES EPITHELIAL CELL ADHESION MOLECULE (EPCAM) AND PLURIPOTENCY GENES. ALSO, KNOCKDOWN OF DDX5 ENHANCED TRANSCRIPTION FROM THE HBV MINICHROMOSOME. THE HELICASE ACTIVITY OF DDX5 STABILIZED SUZ12- AND PRC2-MEDIATED GENE SILENCING, BY DISPLACING THE RNA-BINDING E3 LIGASE, MEX-3 RNA-BINDING FAMILY MEMBER B (MEX3B), FROM HOTAIR. CONVERSELY, ECTOPIC EXPRESSION OF MEX3B UBIQUITINATED SUZ12, DISPLACED DDX5 FROM HOTAIR, AND INDUCED SUZ12 DOWN-REGULATION. IN G2 PHASE OF CELLS EXPRESSING THE HBV X PROTEIN (HBX), SUZ12 PREFERENTIALLY ASSOCIATED WITH MEX3B, BUT NOT DDX5, RESULTING IN DE-REPRESSION OF PRC2 TARGETS, INCLUDING EPCAM AND PLURIPOTENCY GENES. SIGNIFICANTLY, LIVER TUMORS FROM HBX/C-MYC BITRANSGENIC MICE AND CHRONICALLY HBV-INFECTED PATIENTS EXHIBITED A STRONG NEGATIVE CORRELATION BETWEEN DDX5 MESSENGER RNA LEVELS, PLURIPOTENCY GENE EXPRESSION, AND LIVER TUMOR DIFFERENTIATION. NOTABLY, CHRONICALLY INFECTED HBV PATIENTS WITH HCC EXPRESSING REDUCED DDX5 EXHIBITED POOR PROGNOSIS AFTER TUMOR RESECTION, IDENTIFYING DDX5 AS AN IMPORTANT PLAYER IN POOR PROGNOSIS HCC. CONCLUSION: THE RNA HELICASE DDX5, AND E3 LIGASE MEX3B, ARE IMPORTANT CELLULAR TARGETS FOR THE DESIGN OF NOVEL, EPIGENETIC THERAPIES TO COMBAT HBV INFECTION AND POOR PROGNOSIS HBV-ASSOCIATED LIVER CANCER. (HEPATOLOGY 2016;64:1033-1048).	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                 
2 1806  42 EFFECT OF TRANSCRIPTION INHIBITION AND GENERATION OF SUPPRESSIVE VIRAL NON-CODING RNAS. BACKGROUND: HIV-1 PATIENTS RECEIVING COMBINATION ANTIRETROVIRAL THERAPY (CART) SURVIVE INFECTION BUT REQUIRE LIFE-LONG ADHERENCE AT HIGH EXPENSE. IN CHRONIC CART-TREATED PATIENTS WITH UNDETECTABLE VIRAL TITERS, CELL-ASSOCIATED VIRAL RNA IS STILL DETECTABLE, POINTING TO LOW-LEVEL VIRAL TRANSCRIPTIONAL LEAKINESS. TO DATE, THERE ARE NO FDA-APPROVED DRUGS AGAINST HIV-1 TRANSCRIPTION. WE HAVE PREVIOUSLY SHOWN THAT F07#13, A THIRD GENERATION TAT PEPTIDE MIMETIC WITH COMPETITIVE ACTIVITY AGAINST CDK9/T1-TAT BINDING SITES, INHIBITS HIV-1 TRANSCRIPTION IN VITRO AND IN VIVO. RESULTS: HERE, WE DEMONSTRATE THAT INCREASING CONCENTRATIONS OF F07#13 (0.01, 0.1, 1 MICROM) CAUSE A DECREASE IN TAT LEVELS IN A DOSE-DEPENDENT MANNER BY INHIBITING THE CDK9/T1-TAT COMPLEX FORMATION AND SUBSEQUENT UBIQUITIN-MEDIATED TAT SEQUESTRATION AND DEGRADATION. OUR DATA INDICATE THAT COMPLEXES I AND IV CONTAIN DISTINCT PATTERNS OF UBIQUITINATED TAT AND THAT TRANSCRIPTIONAL INHIBITION INDUCED BY F07#13 CAUSES AN OVERALL REDUCTION IN TAT LEVELS. THIS REDUCTION MAY BE TRIGGERED BY F07#13 BUT ULTIMATELY IS MEDIATED BY TAR-GAG VIRAL RNAS THAT BIND SUPPRESSIVE TRANSCRIPTION FACTORS (SIMILAR TO 7SK, NRON, HOTAIR, AND XIST LNCRNAS) TO ENHANCE TRANSCRIPTIONAL GENE SILENCING AND LATENCY. THESE RNAS COMPLEX WITH PRC2, SIN3A, AND CUL4B, RESULTING IN EPIGENETIC MODIFICATIONS. FINALLY, WE OBSERVED AN F07#13-MEDIATED DECREASE OF VIRAL BURDEN BY TARGETING THE R REGION OF THE LONG TERMINAL REPEAT (HIV-1 PROMOTER REGION, LTR), PROMOTING BOTH PAUSED POLYMERASES AND INCREASED EFFICIENCY OF CRISPR/CAS9 EDITING IN INFECTED CELLS. THIS IMPLIES THAT GENE EDITING MAY BE BEST PERFORMED UNDER A REPRESSED TRANSCRIPTIONAL STATE. CONCLUSIONS: COLLECTIVELY, OUR RESULTS INDICATE THAT F07#13, WHICH CAN TERMINATE RNA POLYMERASE II AT DISTINCT SITES, CAN GENERATE SCAFFOLD RNAS, WHICH MAY ASSEMBLE INTO SPECIFIC SETS OF "RNA MACHINES" THAT CONTRIBUTE TO GENE REGULATION. IT REMAINS TO BE SEEN WHETHER THESE EFFECTS CAN ALSO BE SEEN IN VARIOUS CLADES THAT HAVE VARYING PROMOTER STRENGTH, MUTANT LTRS, AND IN PATIENT SAMPLES.	2019	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
3 2316  42 EPIGENETIC REGULATION OF FRUCTOSE-1,6-BISPHOSPHATASE 1 BY HOST TRANSCRIPTION FACTOR SPECKLED 110 KDA DURING HEPATITIS B VIRUS INFECTION. HEPATITIS B VIRUS (HBV) IS THE LEADING CAUSE OF LIVER DISEASE RANGING FROM ACUTE AND CHRONIC HEPATITIS TO LIVER CIRRHOSIS AND HEPATOCELLULAR CARCINOMA (HCC). STUDIES HAVE REVEALED THAT HBV INFECTION BROADLY REPROGRAMMES THE HOST CELLULAR METABOLIC PROCESSES FOR VIRAL PATHOGENESIS. PREVIOUS REPORTS HAVE SHOWN THAT GLYCOLYSIS AND GLUCONEOGENESIS ARE AMONG THE MOST DEREGULATED PATHWAYS DURING HBV INFECTION. WE NOTED THAT DESPITE BEING ONE OF THE RATE-LIMITING ENZYMES OF GLUCONEOGENESIS, THE ROLE AND REGULATION OF FRUCTOSE-1,6-BISPHOSPHATASE 1 (FBP1) DURING HBV INFECTION IS NOT MUCH EXPLORED. IN THIS STUDY, WE REPORT FBP1 UPREGULATION UPON HBV INFECTION AND UNRAVEL A NOVEL MECHANISM OF EPIGENETIC REPROGRAMMING OF FBP1 BY HBV VIA UTILIZING HOST FACTOR SPECKLED 110 KDA (SP110). HERE, WE IDENTIFIED ACETYLATED LYSINE 18 OF HISTONE H3 (H3K18AC) AS A SELECTIVE INTERACTOR OF SP110 BROMODOMAIN. FURTHERMORE, WE FOUND THAT SP110 GETS RECRUITED ON H3K18AC-ENRICHED FBP1 PROMOTER, AND FACILITATES RECRUITMENT OF DEACETYLASE SIRTUIN 2 (SIRT2) ON THAT SITE IN THE PRESENCE OF HBV. SIRT2 IN TURN BRINGS ITS INTERACTOR AND TRANSCRIPTIONAL ACTIVATOR HEPATOCYTE NUCLEAR FACTOR 4-ALPHA TO THE PROMOTER, WHICH ULTIMATELY LEADS TO A LOSS OF DNA METHYLATION NEAR THE COGNATE SITE. INTERESTINGLY, THIS SP110 DRIVEN FBP1 REGULATION DURING INFECTION WAS FOUND TO PROMOTE VIRAL-BORNE HCC PROGRESSION. MOREOVER, SP110 CAN BE USED AS A PROGNOSTIC MARKER FOR THE HEPATITIS-MEDIATED HCC PATIENTS, WHERE HIGH SP110 EXPRESSION SIGNIFICANTLY LOWERED THEIR SURVIVAL. THUS, THE EPIGENETIC READER PROTEIN SP110 HAS POTENTIAL TO BE A THERAPEUTIC TARGET TO CHALLENGE HBV-INDUCED HCCS.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              
4 5678  40 SHORT HAIRPIN RNA INDUCES METHYLATION OF HEPATITIS B VIRUS COVALENTLY CLOSED CIRCULAR DNA IN HUMAN HEPATOMA CELLS. SMALL INTERFERING RNAS NOT ONLY MODULATE GENE EXPRESSION AT A POST-TRANSCRIPTIONAL LEVEL, BUT ALSO INDUCE TRANSCRIPTIONAL GENE SILENCING BY RNA INTERFERENCE-MEDIATED HETEROCHROMATIN FORMATION AND RNA-DIRECTED DNA METHYLATION (RDDM). HOWEVER, ALTHOUGH ESTABLISHED IN PLANTS, THERE HAVE BEEN CONTROVERSIES WHETHER RDDM OPERATES IN MAMMALS. HEPATITIS B VIRUS (HBV) COVALENTLY CLOSED CIRCULAR DNA (CCCDNA) SERVES AS A TEMPLATE FOR VIRAL RNA TRANSCRIPTION, AND TRANSCRIPTIONAL ACTIVITY OF HBV CCCDNA IS REGULATED BY METHYLATION IN PATIENTS WITH CHRONIC HBV INFECTION. IN THIS STUDY, WE STABLY EXPRESSED SHORT HAIRPIN RNA (SHRNA) AGAINST HBV IN HUMAN HEPATOMA CELLS TO DETERMINE WHETHER SHRNA INDUCES METHYLATION OF HBV CCCDNA. HEPAD38 CELLS WHICH PERMIT REPLICATION OF HBV UNDER CONTROL OF TETRACYCLINE-RESPONSIVE PROMOTER WERE TRANSDUCED WITH LENTIVIRAL VECTORS WHICH ENCODE SH-1580, A SHRNA AGAINST THE HEPATITIS B VIRAL PROTEIN HBX. BISULFITE SEQUENCING PCR ANALYSIS REVEALED THAT SH-1580 INDUCED CPG METHYLATIONS AT A HIGHER RATE COMPARED TO CONTROL (31.3% VS. 12.8%, P<0.05). THE SH-1580-INDUCED CPG METHYLATION WAS LOCALIZED NEAR THE TARGET SEQUENCE OF SH-1580 IN MORE THAN A HALF OF THE CLONES. METHYLATION-INDUCED TRANSCRIPTIONAL SUPPRESSION WAS CONFIRMED BY IN VITRO TRANSCRIPTION ASSAY. THESE RESULTS CONFIRM THE FEASIBILITY OF RDDM OF HBV CCCDNA IN HUMAN CELLS. LENTIVIRAL VECTOR-MEDIATED TRANSFER OF SHRNA MAY BE USED AS A TOOL FOR NOVEL TRANSCRIPTIONAL MODULATION BY EPIGENETIC MODIFICATION OF HBV CCCDNA.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
5 5101  33 POLYCOMB FACTOR PHF19 CONTROLS CELL GROWTH AND DIFFERENTIATION TOWARD ERYTHROID PATHWAY IN CHRONIC MYELOID LEUKEMIA CELLS. POLYCOMB GROUP (PCG) OF PROTEINS ARE A GROUP OF HIGHLY CONSERVED EPIGENETIC REGULATORS INVOLVED IN MANY BIOLOGICAL FUNCTIONS, SUCH AS EMBRYONIC DEVELOPMENT, CELL PROLIFERATION, AND ADULT STEM CELL DETERMINATION. PHD FINGER PROTEIN 19 (PHF19) IS AN ASSOCIATED FACTOR OF POLYCOMB REPRESSOR COMPLEX 2 (PRC2), OFTEN UPREGULATED IN HUMAN CANCERS. IN PARTICULAR, MYELOID LEUKEMIA CELL LINES SHOW INCREASED LEVELS OF PHF19, YET LITTLE IS KNOWN ABOUT ITS FUNCTION. HERE, WE HAVE CHARACTERIZED THE ROLE OF PHF19 IN MYELOID LEUKEMIA CELLS. WE DEMONSTRATED THAT PHF19 DEPLETION DECREASES CELL PROLIFERATION AND PROMOTES CHRONIC MYELOID LEUKEMIA (CML) DIFFERENTIATION. MECHANISTICALLY, WE HAVE SHOWN HOW PHF19 REGULATES THE PROLIFERATION OF CML THROUGH A DIRECT REGULATION OF THE CELL CYCLE INHIBITOR P21. FURTHERMORE, WE OBSERVED THAT MTF2, A PHF19 HOMOLOG, PARTIALLY COMPENSATES FOR PHF19 DEPLETION IN A SUBSET OF TARGET GENES, INSTRUCTING SPECIFIC ERYTHROID DIFFERENTIATION. TAKEN TOGETHER, OUR RESULTS SHOW THAT PHF19 IS A KEY TRANSCRIPTIONAL REGULATOR FOR CELL FATE DETERMINATION AND COULD BE A POTENTIAL THERAPEUTIC TARGET FOR MYELOID LEUKEMIA TREATMENT.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
6 3128  40 GIPC-REGULATED IGFBP-3 PROMOTES HSC MIGRATION IN VITRO AND PORTAL HYPERTENSION IN VIVO THROUGH A BETA1-INTEGRIN PATHWAY. BACKGROUND & AIMS: TRANSFORMING GROWTH FACTOR (TGF-BETA)-INDUCED ACTIVATION OF QUIESCENT HEPATIC STELLATE CELLS (HSCS) AND THEIR TRANSFORMATION TO MYOFIBROBLASTS IS A KEY EVENT IN LIVER FIBROSIS AND PORTAL HYPERTENSION. GIPC (ALSO REFERRED TO AS SYNECTIN) IS A DOWNSTREAM SIGNAL ACTIVATION MOLECULE OF TGF-BETA AND OTHER RECEPTORS. IN THIS STUDY, WE SOUGHT TO IDENTIFY NOVEL GENES TARGETED BY TGF-BETA AND GIPC AND ELUCIDATE IF AND HOW THEY MAY CONTRIBUTE TO LIVER FIBROSIS. METHODS: WE PERFORMED SEQUENTIAL MESSENGER RNA SEQUENCING ANALYSIS ON TGF-BETA-STIMULATED HSCS AND THEN ON TGF-BETA-STIMULATED HSCS IN THE PRESENCE AND ABSENCE OF GIPC ALSO REFERRED TO AS SYNECTIN (GIPC) KNOCKDOWN. INSULIN-LIKE GROWTH FACTOR BINDING PROTEIN-3 (IGFBP-3) TRANSPORT PROTEIN EMERGED AS A TOP ACTIVATION TARGET OF BOTH TGF-BETA AND GIPC. QUANTITATIVE POLYMERASE CHAIN REACTION, ENZYME-LINKED IMMUNOSORBENT ASSAY, TARGETED CHROMATIN IMMUNOPRECIPITATION, AND WESTERN BLOT ANALYSIS WERE DONE FOR FURTHER CONFIRMATION. RESULTS: IGFBP-3, AN INSULIN GROWTH FACTOR TRANSPORT PROTEIN, EMERGED AS A TOP ACTIVATION TARGET OF BOTH TGF-BETA AND GIPC, WHICH WAS CONFIRMED BY QUANTITATIVE POLYMERASE CHAIN REACTION, ENZYME-LINKED IMMUNOSORBENT ASSAY, AND WESTERN BLOT ANALYSIS. TARGETED CHROMATIN IMMUNOPRECIPITATION SHOWED THAT GIPC INCREASES THE HISTONE 3 LYSINE 27 (H3K27) ACETYLATION ACTIVATING MARK AND CONCURRENTLY DECREASES THE H3K27 INHIBITORY TRIMETHYLATION (H3K27M3) MARK, PROVIDING AN EPIGENETIC CORRELATE TO THE GENE REGULATION CHANGES. IN VIVO, GLOBAL KNOCKOUT OF IGFBP-3 MICE RESULTED IN ATTENUATION OF HSC ACTIVATION MARKERS AND ATTENUATION OF PORTAL PRESSURE IN RESPONSE TO CHRONIC LIVER INJURY MODELS. ANALYSIS OF SERUM LEVELS FROM CIRRHOTIC PATIENTS ALSO SHOWED AN IGFBP-3 INCREASE OF MORE THAN 2-FOLD COMPARED WITH HEALTHY CONTROLS. FINALLY, IN VITRO MECHANISM STUDIES SHOWED THAT IGFBP-3 PROMOTES HSC MIGRATION THROUGH INTEGRIN-DEPENDENT PHOSPHORYLATION OF PROTEIN KINASE B. CONCLUSIONS: TGF-BETA UP-REGULATES IGFBP-3 THROUGH GIPC, LEADING TO INCREASED HSC MIGRATION IN VITRO AND PROMOTES PORTAL HYPERTENSION IN VIVO. THESE STUDIES SUPPORT THE ROLE OF IGFBP-3 AS A POTENTIAL PATHOPHYSIOLOGIC TARGET OR BIOMARKER IN CHRONIC LIVER DISEASE.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
7 2432  39 EPIGENETIC SILENCING OF MIR-708 ENHANCES NF-KAPPAB SIGNALING IN CHRONIC LYMPHOCYTIC LEUKEMIA. MICRORNAS (MIRNAS) ARE POST-TRANSCRIPTIONAL REGULATORS OF GENE EXPRESSION AND THEIR DEREGULATION IS INVOLVED IN TUMOR DEVELOPMENT. EPIGENETIC GENE SILENCING IN CANCER BY DNA METHYLATION CONTRIBUTES TO THE SILENCING OF TUMOR-SUPPRESSOR GENES, INCLUDING MIRNAS. WE HAVE RECENTLY SHOWN THAT THE PROMOTER OF MIR-708 IS ABERRANTLY METHYLATED IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). TO CHARACTERIZE THE MOLECULAR SIGNALING NETWORKS THAT ARE INFLUENCED BY MIR-708, WE PERFORMED A LUCIFERASE-BASED SCREEN EVALUATING THE EFFECTS OF ECTOPIC MIR-708 EXPRESSION ON LEUKEMIA-RELEVANT SIGNALING PATHWAYS. WE FOUND THAT MIR-708 STRONGLY REPRESSED NF-KAPPAB SIGNALING, A PATHWAY KNOWN TO BE DEREGULATED IN CLL. AMONG THE PREDICTED MIR-708 TARGETS WAS IKKBETA (INHIBITOR OF KAPPA LIGHT POLYPEPTIDE GENE ENHANCER IN B CELLS, KINASE-BETA/IKBKB), A KEY KINASE FACILITATING NF-KAPPAB SIGNALING. WE VALIDATED THE INTERACTION OF MIR-708 WITH THE 3'-UNTRANSLATED REGION OF IKKBETA AND FOUND THAT MIR-708 OVEREXPRESSION REPRESSES ENDOGENOUS IKKBETA. PHOSPHORYLATION OF THE IKKBETA TARGET IKAPPABALPHA AND EXPRESSION OF KNOWN NF-KAPPAB TARGET GENES WERE IMPAIRED BY MIR-708. FURTHERMORE, WE IDENTIFIED AN ENHANCER REGION DOWNSTREAM OF THE MIR-708 PROMOTER THAT DISPLAYS A DISTINCT DNA METHYLATION STATUS IN CLL. HIGH ENHANCER METHYLATION IS SIGNIFICANTLY CORRELATED WITH LOWER MIR-708 EXPRESSION AND IS PREDOMINANTLY FOUND IN PATIENTS WITH POOR PROGNOSIS AND SHORTER TIME TO TREATMENT. THESE RESULTS DEMONSTRATE THAT MIR-708 REGULATES THE NF-KAPPAB PATHWAY BY TARGETING IKKBETA, AND THAT METHYLATION OF A KEY ENHANCER REGION CONTRIBUTES TO ITS SUPPRESSION IN CLL.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
8 1334  31 DEREGULATION OF AIOLOS EXPRESSION IN CHRONIC LYMPHOCYTIC LEUKEMIA IS ASSOCIATED WITH EPIGENETIC MODIFICATIONS. CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS CHARACTERIZED BY A CLONAL ACCUMULATION OF MATURE NEOPLASTIC B CELLS THAT ARE RESISTANT TO APOPTOSIS. AIOLOS, A MEMBER OF THE IKAROS FAMILY OF ZINC-FINGER TRANSCRIPTION FACTORS, PLAYS AN IMPORTANT ROLE IN THE CONTROL OF MATURE B LYMPHOCYTE DIFFERENTIATION AND MATURATION. IN THIS STUDY, WE SHOWED THAT AIOLOS EXPRESSION IS UP-REGULATED IN B-CLL CELLS. THIS OVEREXPRESSION DOES NOT IMPLICATE ISOFORM IMBALANCE OR DISTURB AIOLOS SUBCELLULAR LOCALIZATION. THE CHROMATIN STATUS AT THE AIOLOS PROMOTER IN CLL IS DEFINED BY THE DEMETHYLATION OF DNA AND AN ENRICHMENT OF EUCHROMATIN ASSOCIATED HISTONE MARKERS, SUCH AS THE DIMETHYLATION OF THE LYSINE 4 ON HISTONE H3. THESE EPIGENETIC MODIFICATIONS SHOULD ALLOW ITS UPSTREAM EFFECTORS, SUCH AS NUCLEAR FACTOR-KAPPAB, CONSTITUTIVELY ACTIVATED IN CLL, TO GAIN ACCESS TO PROMOTER, RESULTING UP-REGULATION OF AIOLOS. TO DETERMINE THE CONSEQUENCES OF AIOLOS DEREGULATION IN CLL, WE ANALYZED THE EFFECTS OF AIOLOS OVEREXPRESSION OR DOWN-REGULATION ON APOPTOSIS. AIOLOS IS INVOLVED IN CELL SURVIVAL BY REGULATING THE EXPRESSION OF SOME BCL-2 FAMILY MEMBERS. OUR RESULTS STRONGLY SUGGEST THAT AIOLOS DEREGULATION BY EPIGENETIC MODIFICATIONS MAY BE A HALLMARK OF CLL.	2011	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
9 4696  40 NF-KAPPAB REPRESSES RETINOIC ACID RECEPTOR-MEDIATED GPRC5A TRANSACTIVATION IN LUNG EPITHELIAL CELLS TO PROMOTE NEOPLASIA. CHRONIC INFLAMMATION IS ASSOCIATED WITH LUNG TUMORIGENESIS, IN WHICH NF-KAPPAB-MEDIATED EPIGENETIC REGULATION PLAYS A CRITICAL ROLE. LUNG TUMOR SUPPRESSOR G PROTEIN-COUPLED RECEPTOR, FAMILY C, MEMBER 5A (GPRC5A), IS REPRESSED IN MOST NON-SMALL CELL LUNG CANCER (NSCLC); HOWEVER, THE MECHANISMS REMAIN UNCLEAR. HERE, WE SHOW THAT NF-KAPPAB ACTS AS A TRANSCRIPTIONAL REPRESSOR IN SUPPRESSION OF GPRC5A. NF-KAPPAB INDUCED GPRC5A REPRESSION BOTH IN VITRO AND IN VIVO. INTRIGUINGLY, TRANSACTIVATION OF NF-KAPPAB DOWNSTREAM TARGETS WAS NOT REQUIRED, BUT THE TRANSACTIVATION DOMAIN OF RELA/P65 WAS REQUIRED FOR GPRC5A REPRESSION. NF-KAPPAB DID NOT BIND TO ANY POTENTIAL CIS-ELEMENT IN THE GPRC5A PROMOTER. INSTEAD, P65 WAS COMPLEXED WITH RETINOIC ACID RECEPTOR ALPHA/BETA (RARALPHA/BETA) AND RECRUITED TO THE RA RESPONSE ELEMENT SITE AT THE GPRC5A PROMOTER, RESULTING IN DISRUPTED RNA POLYMERASE II COMPLEXING AND SUPPRESSED TRANSCRIPTION. NOTABLY, PHOSPHORYLATION ON SERINE 276 OF P65 WAS REQUIRED FOR INTERACTION WITH RARALPHA/BETA AND REPRESSION OF GPRC5A. MOREOVER, NF-KAPPAB-MEDIATED EPIGENETIC REPRESSION WAS THROUGH SUPPRESSION OF ACETYLATED HISTONE H3K9 (H3K9AC), BUT NOT DNA METHYLATION OF THE CPG ISLANDS, AT THE GPRC5A PROMOTER. CONSISTENTLY, A HISTONE DEACETYLASE INHIBITOR, BUT NOT DNA METHYLATION INHIBITOR, RESTORED GPRC5A EXPRESSION IN NSCLC CELLS. THUS, NF-KAPPAB INDUCES TRANSCRIPTIONAL REPRESSION OF GPRC5A VIA A COMPLEX WITH RARALPHA/BETA AND MEDIATES EPIGENETIC REPRESSION VIA SUPPRESSION OF H3K9AC.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
10 6659  39 UPREGULATION OF AKT3 CONFERS RESISTANCE TO THE AKT INHIBITOR MK2206 IN BREAST CANCER. ACQUIRED RESISTANCE TO MOLECULAR TARGETED THERAPY REPRESENTS A MAJOR CHALLENGE FOR THE EFFECTIVE TREATMENT OF CANCER. HYPERACTIVATION OF THE PI3K/AKT PATHWAY IS FREQUENTLY OBSERVED IN VIRTUALLY ALL HUMAN MALIGNANCIES, AND NUMEROUS PI3K AND AKT INHIBITORS ARE CURRENTLY UNDER CLINICAL EVALUATION. HOWEVER, MECHANISMS OF ACQUIRED RESISTANCE TO AKT INHIBITORS HAVE YET TO BE DESCRIBED. HERE, WE USE A BREAST CANCER PRECLINICAL MODEL TO IDENTIFY RESISTANCE MECHANISMS TO A SMALL MOLECULE ALLOSTERIC AKT INHIBITOR, MK2206. USING A STEP-WISE AND CHRONIC HIGH-DOSE EXPOSURE, BREAST CANCER CELL LINES HARBORING ONCOGENIC PI3K RESISTANT TO MK2206 WERE ESTABLISHED. USING THIS MODEL, WE REVEAL THAT AKT3 EXPRESSION IS MARKEDLY UPREGULATED IN AKT INHIBITOR-RESISTANT CELLS. INDUCTION OF AKT3 IS REGULATED EPIGENETICALLY BY THE BROMODOMAIN AND EXTRA TERMINAL DOMAIN PROTEINS. IMPORTANTLY, KNOCKDOWN OF AKT3, BUT NOT AKT1 OR AKT2, IN RESISTANT CELLS RESTORES SENSITIVITY TO MK2206. AKT INHIBITOR-RESISTANT CELLS ALSO DISPLAY AN EPITHELIAL TO MESENCHYMAL TRANSITION PHENOTYPE AS ASSESSED BY ALTERATIONS IN THE LEVELS OF E-CADHERIN, N-CADHERIN, AND VIMENTIN, AS WELL AS ENHANCED INVASIVENESS OF TUMOR SPHEROIDS. NOTABLY, THE INVASIVE MORPHOLOGY OF RESISTANT SPHEROIDS IS DIMINISHED UPON AKT3 DEPLETION. WE ALSO SHOW THAT RESISTANCE TO MK2206 IS REVERSIBLE BECAUSE UPON DRUG REMOVAL RESISTANT CELLS REGAIN SENSITIVITY TO AKT INHIBITION, ACCOMPANIED BY REEXPRESSION OF EPITHELIAL MARKERS AND REDUCTION OF AKT3 EXPRESSION, IMPLYING THAT EPIGENETIC REPROGRAMMING CONTRIBUTES TO ACQUISITION OF RESISTANCE. THESE FINDINGS PROVIDE A RATIONALE FOR DEVELOPING THERAPEUTICS TARGETING AKT3 TO CIRCUMVENT ACQUIRED RESISTANCE IN BREAST CANCER. MOL CANCER THER; 15(8); 1964-74. (C)2016 AACR.	2016	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
11 6385  46 THE ROLE OF PROMYELOCYTIC LEUKEMIA PROTEIN IN STEATOSIS-ASSOCIATED HEPATIC TUMORS RELATED TO CHRONIC HEPATITIS B VIRUS INFECTION. THE PERSISTENCE OF HEPATITIS B SURFACE ANTIGEN (HBSAG) IS A RISK FACTOR FOR THE DEVELOPMENT OF STEATOSIS-ASSOCIATED TUMORS IN CHRONIC HEPATITIS B VIRUS (HBV) INFECTION, YET LITTLE IS KNOWN ABOUT THE METABOLIC LINK WITH THIS FACTOR. WE CORRELATED HBV-RELATED PATHOGENESIS IN GENETICALLY ENGINEERED MICE AND HUMAN CARRIERS WITH METABOLIC PROTEOMICS AND LIPOGENIC GENE EXPRESSION PROFILES. THE IMMUNOHISTOCHEMISTRY SHOWED THAT THE PROMYELOCYTIC LEUKEMIA PROTEIN (PML, A TUMOR SUPPRESSOR INVOLVED IN GENOME MAINTENANCE AND FATTY ACID OXIDATION), BEING INVERSELY INFLUENCED BY THE DYNAMIC HBSAG LEVELS FROM ACUTE PHASE TO SEROCLEARANCE, APPEARED AS A LIPO-METABOLIC SWITCH LINKING HBSAG-INDUCED STEATOSIS (LIPOGENESIS) TO HBSAG-LOST FAT-BURNING HEPATOCARCINOGENESIS (LIPOLYSIS). KNOCKDOWN OF PML IN HBSAG-TRANSGENIC MICE PREDISPOSED TO OBESITY AND DROVE EARLY STEATOSIS-SPECIFIC LIVER TUMORIGENESIS. PROTEOME ANALYSIS REVEALED THAT THE SIGNALING PATHWAYS CORRESPONDING TO ENERGY METABOLISM AND ITS REGULATORS WERE FREQUENTLY ALTERED BY SUPPRESSION OR DEPLETION OF PML IN THE HBSAG-TRANSGENIC MICE, MAINLY INCLUDING OXIDATIVE PHOSPHORYLATION AND FATTY ACID METABOLISM. EXPRESSION PROFILING FURTHER IDENTIFIED UPREGULATION OF STEAROYL-COA DESATURASE 1 (SCD1) AND EPIGENETIC METHYLATION OF NDUFA13 IN THE MITOCHONDRIAL RESPIRATORY CHAIN AND THE CELL CYCLE INHIBITOR CDKN1C IN CONCORDANCE TO THE INCREASED SEVERITY OF LIPODYSTROPHY AND NEOPLASIA IN THE LIVERS OF HBSAG-TRANSGENIC MICE WITH PML INSUFFICIENCY. THE DEFECT IN LIPOLYSIS IN PML-DEFICIENT HBSAG-TRANSGENIC MICE MADE THE HBSAG-INDUCED ADIPOSE-LIKE LIVER TUMORS VULNERABLE TO SYNTHETIC LETHALITY FROM TOXIC SATURATED FAT ACCUMULATION WITH A SCD1 INHIBITOR. OUR FINDINGS PROVIDE MECHANISTIC INSIGHTS INTO THE EVOLUTION OF STEATOSIS-ASSOCIATED HEPATIC TUMORS DRIVEN BY RECIPROCAL INTERACTIONS OF HBSAG AND PML, AND A POTENTIAL UTILITY OF LIPID METABOLIC REPROGRAMMING AS A TREATMENT TARGET.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          
12 5004  32 PERIPHERAL B CELLS FROM PATIENTS WITH HEPATITIS C VIRUS-ASSOCIATED LYMPHOMA EXHIBIT CLONAL EXPANSION AND AN ANERGIC-LIKE TRANSCRIPTIONAL PROFILE. CHRONIC HCV INFECTION REMAINS A GLOBAL HEALTH CONCERN DUE TO ITS INVOLVEMENT IN HEPATIC AND EXTRAHEPATIC DISEASES, INCLUDING B CELL NON-HODGKIN LYMPHOMA (BNHL). CLINICAL AND EPIDEMIOLOGICAL EVIDENCE SUPPORT A CAUSAL ROLE FOR HCV IN BNHL DEVELOPMENT, ALTHOUGH MECHANISTIC INSIGHT IS LACKING. WE PERFORMED RNA-SEQUENCING ON PERIPHERAL B CELLS FROM PATIENTS WITH HCV ALONE, BNHL ALONE, AND HCV-ASSOCIATED BNHL TO IDENTIFY UNIQUE AND SHARED TRANSCRIPTIONAL PROFILES ASSOCIATED WITH TRANSFORMATION. IN PATIENTS WITH HCV-ASSOCIATED BNHL, WE OBSERVED THE ENRICHMENT OF AN ANERGIC-LIKE GENE SIGNATURE AND EVIDENCE OF CLONAL EXPANSION THAT WAS CORRELATED WITH THE EXPRESSION OF EPIGENETIC REGULATORY GENES. OUR DATA SUPPORT A ROLE FOR VIRAL-MEDIATED CLONAL EXPANSION OF ANERGIC-LIKE B CELLS IN HCV-ASSOCIATED BNHL DEVELOPMENT AND SUGGEST EPIGENETIC DYSREGULATION AS A POTENTIAL MECHANISM DRIVING EXPANSION. WE PROPOSE EPIGENETIC MECHANISMS MAY BE INVOLVED IN BOTH HCV-ASSOCIATED LYMPHOMA AND REGULATION OF B CELL ANERGY, REPRESENTING AN ATTRACTIVE TARGET FOR CLINICAL INTERVENTIONS.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
13 3249  36 HEPATITIS B VIRUS HIJACKS CTHRC1 TO EVADE HOST IMMUNITY AND MAINTAIN REPLICATION. HEPATITIS B VIRUS (HBV) INFECTION CAUSES ACUTE AND CHRONIC LIVER DISEASES, BUT IS NOT DIRECTLY CYTOPATHIC. LIVER INJURY RESULTS FROM REPEATED ATTEMPTS OF THE CELLULAR IMMUNE RESPONSE SYSTEM TO CONTROL THE VIRAL INFECTION. HERE, WE INVESTIGATE THE ROLES OF CELLULAR FACTORS AND SIGNALING PATHWAYS INVOLVED IN THE REGULATION OF HBV REPLICATION TO REVEAL THE MECHANISM UNDERLYING HBV INFECTION AND PATHOGENESIS. WE SHOW THAT COLLAGEN TRIPLE HELIX REPEAT CONTAINING 1 (CTHRC1) EXPRESSION IS ELEVATED IN HBV-INFECTED PATIENTS AND IN HBV-TRANSFECTED CELLS THROUGH EPIGENETIC MODIFICATION AND TRANSCRIPTIONAL REGULATION. CTHRC1 FACILITATES HBV REPLICATION IN CULTURED CELLS AND BALB/C MICE BY ACTIVATING THE PKCALPHA/ERK/JNK/C-JUN CASCADE TO REPRESS THE IFN/JAK/STAT PATHWAY. HBV-ACTIVATED CTHRC1 DOWNREGULATES THE ACTIVITY OF TYPE I INTERFERON (IFN), THE PRODUCTION OF IFN-STIMULATED GENES (ISGS), AND THE PHOSPHORYLATION OF SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION 1/2 (STAT1/2), WHEREAS IT UPREGULATES THE PHOSPHORYLATION AND UBIQUITINATION OF TYPE I IFN RECEPTORS (IFNARALPHA/BETA). THUS, OUR RESULTS SHOW THAT HBV USES A NOVEL MECHANISM TO HIJACK CELLULAR FACTORS AND SIGNAL CASCADES IN ORDER TO EVADE HOST ANTIVIRAL IMMUNITY AND MAINTAIN PERSISTENT INFECTION. WE ALSO DEMONSTRATE THAT CTHRC1 HAS A NOVEL ROLE IN VIRAL INFECTION.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
14 1067  43 CLINICAL UTILITY OF PDSS2 EXPRESSION TO STRATIFY PATIENTS AT RISK FOR RECURRENCE OF HEPATOCELLULAR CARCINOMA. IDENTIFICATION OF NOVEL GENETIC AND EPIGENETIC ALTERATIONS IS REQUIRED FOR OPTIMAL STRATIFICATION OF PATIENTS WITH HEPATOCELLULAR CARCINOMA (HCC) AT RISK FOR RECURRENCE AND ADVERSE PROGNOSIS. COENZYME Q10 (COQ10), WHICH MEDIATES APOPTOSIS, IS SYNTHESIZED BY PRENYL DIPHOSPHATE SYNTHASE SUBUNIT 2 (PDSS2). IN THE PRESENT STUDY WE EVALUATED THE CLINICAL SIGNIFICANCE AND REGULATORY MECHANISMS OF PDSS2 EXPRESSION IN HCC. PDSS2 EXPRESSION LEVELS AND THOSE OF GENES ENCODING POTENTIALLY INTERACTING PROTEINS AS WELL AS THE METHYLATION STATUS OF THE PDSS2 PROMOTER REGION WERE ANALYZED IN HCC CELL LINES. PDSS2 MRNA LEVELS IN 151 PAIRS OF RESECTED SPECIMENS WERE DETERMINED TO EVALUATE THE ASSOCIATION OF PDSS2 EXPRESSION AND CLINICOPATHOLOGICAL FACTORS. THE EXPRESSION AND DISTRIBUTION OF PDSS2 WERE DETERMINED USING IMMUNOHISTOCHEMISTRY. PDSS2 MRNA EXPRESSION WAS DECREASED IN SIX OF NINE HCC CELL LINES AND SIGNIFICANTLY CORRELATED WITH THOSE OF HEPATOCYTE NUCLEAR FACTOR 4ALPHA. PDSS2 TRANSCRIPTION IN HCC CELLS WITH DECREASED PDSS2 EXPRESSION ACCOMPANYING HYPERMETHYLATION WAS REACTIVATED AFTER TREATING THESE CELLS WITH A METHYLATION INHIBITOR. MEAN EXPRESSION LEVELS OF PDSS2 MRNA RELATIVE TO THAT OF UNINVOLVED LIVER DIMINISHED GRADUALLY IN THE ORDER OF CHRONIC HEPATITIS TO CIRRHOSIS, AND EACH WAS SIGNIFICANTLY HIGHER THAN THOSE OF HCCS. PDSS2 AND PDSS2 MRNA LEVELS WERE CONSISTENT. DECREASED PDSS2 MRNA LEVELS WERE DETECTED IN HCC TISSUES OF 56 PATIENTS, CORRELATED WITH SHORTER DISEASE-SPECIFIC SURVIVAL, AND WAS IDENTIFIED AS AN INDEPENDENT PROGNOSTIC FACTOR. PDSS2 IS A PUTATIVE TUMOR SUPPRESSOR, AND PROMOTER HYPERMETHYLATION IS A KEY REGULATORY MECHANISM IN HCC. DECREASED LEVELS OF PDSS2 MRNA EXPRESSION MAY REPRESENT A NOVEL BIOMARKER OF HCC.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
15 3252  52 HEPATITIS B VIRUS SUPPRESSES COMPLEMENT C9 SYNTHESIS BY LIMITING THE AVAILABILITY OF TRANSCRIPTION FACTOR USF-1 AND INHIBITS FORMATION OF MEMBRANE ATTACK COMPLEX: IMPLICATIONS IN DISEASE PATHOGENESIS. BACKGROUND: THE COMPLEMENT SYSTEM FUNCTIONS PRIMARILY AS A FIRST-LINE HOST DEFENSE AGAINST INVADING MICROBES, INCLUDING VIRUSES. HOWEVER, THE INTERACTION OF HEPATITIS B VIRUS (HBV) WITH THE COMPLEMENT-COMPONENTS DURING CHRONIC HBV INFECTION REMAINS LARGELY UNKNOWN. WE INVESTIGATED THE MECHANISM BY WHICH HBV INHIBITS THE FORMATION OF CYTOLYTIC COMPLEMENT MEMBRANE-ATTACK COMPLEX (MAC) AND STUDIED ITS IMPACT ON MAC-MEDIATED MICROBICIDAL ACTIVITY AND DISEASE PATHOGENESIS. METHODS: BLOOD/LIVER TISSUES WERE COLLECTED FROM CHRONICALLY HBV-INFECTED PATIENTS AND CONTROLS. HEPG2(HNTCP) CELLS WERE INFECTED WITH HBV PARTICLES AND HUH7 CELLS WERE TRANSFECTED WITH FULL-LENGTH LINEAR HBV-MONOMER OR PLASMIDS CONTAINING DIFFERENT HBV-ORFS AND EXPRESSION OF COMPLEMENT COMPONENTS OR OTHER HOST GENES WERE EVALUATED. ADDITIONALLY, ELISA, REAL-TIME PCR, WESTERN BLOT, BIOINFORMATICS ANALYSIS, GENE OVEREXPRESSION/KNOCK-DOWN, MUTAGENESIS, CHROMATIN IMMUNOPRECIPITATION, EPIGENETIC STUDIES, IMMUNOFLUORESCENCE, AND QUANTIFICATION OF SERUM HBV-DNA, BACTERIAL-DNA AND ENDOTOXIN WERE PERFORMED. RESULTS: AMONG THE MAC COMPONENTS (C5B-C9), SIGNIFICANT REDUCTION WAS NOTED IN THE EXPRESSION OF C9, THE MAJOR CONSTITUENT OF MAC, IN HBV-INFECTED HEPG2(HNTCP) CELLS AND IN HUH7 CELLS TRANSFECTED WITH FULL-LENGTH HBV AS WELL AS HBX. C9 LEVEL WAS ALSO MARKED LOW IN SERA/LIVER OF CHRONIC HEPATITIS B (CHB) AND IMMUNE-TOLERANT (IT) PATIENTS THAN INACTIVE CARRIERS AND HEALTHY CONTROLS. HBX STRONGLY REPRESSED C9-PROMOTER ACTIVITY IN HUH7 CELLS BUT CPG-ISLAND WAS NOT DETECTED IN C9-PROMOTER. WE IDENTIFIED USF-1 AS THE KEY TRANSCRIPTION FACTOR THAT DRIVES C9 EXPRESSION AND DEMONSTRATED THAT HBX-INDUCED HYPERMETHYLATION OF USF-1-PROMOTER IS THE LEADING CAUSE OF USF-1 DOWNREGULATION THAT IN TURN DIMINISHED C9 TRANSCRIPTION. REDUCED MAC FORMATION AND IMPAIRED LYSIS OF HBV-TRANSFECTED HUH7 AND BACTERIAL CELLS WERE OBSERVED FOLLOWING INCUBATION OF THESE CELLS WITH C9-DEFICIENT CHB SERA BUT WAS REVERSED UPON C9 SUPPLEMENTATION. SIGNIFICANT INVERSE CORRELATION WAS NOTED BETWEEN C9 CONCENTRATION AND HBV-DNA, BACTERIAL-DNA AND ENDOTOXIN CONTENT IN HBV-INFECTED PATIENTS. ONE-YEAR TENOFOVIR THERAPY RESULTED IN IMPROVEMENT IN C9 LEVEL AND DECLINE IN VIRAL/BACTERIAL/ENDOTOXIN LOAD IN CHB PATIENTS. CONCLUSION: COLLECTIVELY, HBX SUPPRESSED C9 TRANSCRIPTION BY RESTRICTING THE AVAILABILITY OF USF-1 THROUGH HYPERMETHYLATION OF USF-1-PROMOTER AND CONSEQUENTLY HINDER THE FORMATION AND LYTIC FUNCTIONS OF MAC. EARLY THERAPY IS NEEDED FOR BOTH CHB AND IT TO NORMALIZE THE ABERRANT COMPLEMENT PROFILE AND CONTAIN VIRAL AND BACTERIAL INFECTION AND LIMIT DISEASE PROGRESSION.	2022	

16 6448  48 THERAPEUTIC SHUTDOWN OF HBV TRANSCRIPTS PROMOTES REAPPEARANCE OF THE SMC5/6 COMPLEX AND SILENCING OF THE VIRAL GENOME IN VIVO. OBJECTIVE: THERAPEUTIC STRATEGIES SILENCING AND REDUCING THE HEPATITIS B VIRUS (HBV) RESERVOIR, THE COVALENTLY CLOSED CIRCULAR DNA (CCCDNA), HAVE THE POTENTIAL TO CURE CHRONIC HBV INFECTION. WE AIMED TO INVESTIGATE THE IMPACT OF SMALL INTERFERRING RNA (SIRNA) TARGETING ALL HBV TRANSCRIPTS OR PEGYLATED INTERFERON-ALPHA (PEG-IFNALPHA) ON THE VIRAL REGULATORY HBX PROTEIN AND THE STRUCTURAL MAINTENANCE OF CHROMOSOME 5/6 COMPLEX (SMC5/6), A HOST FACTOR SUPPRESSING CCCDNA TRANSCRIPTION. IN PARTICULAR, WE ASSESSED WHETHER INTERVENTIONS LOWERING HBV TRANSCRIPTS CAN ACHIEVE AND MAINTAIN SILENCING OF CCCDNA TRANSCRIPTION IN VIVO. DESIGN: HBV-INFECTED HUMAN LIVER CHIMERIC MICE WERE TREATED WITH SIRNA OR PEG-IFNALPHA. VIROLOGICAL AND HOST CHANGES WERE ANALYSED AT THE END OF TREATMENT AND DURING THE REBOUND PHASE BY QUALITATIVE PCR, ELISA, IMMUNOBLOTTING AND CHROMATIN IMMUNOPRECIPITATION. RNA IN SITU HYBRIDISATION WAS COMBINED WITH IMMUNOFLUORESCENCE TO DETECT SMC6 AND HBV RNAS AT SINGLE CELL LEVEL. THE ENTRY INHIBITOR MYRCLUDEX-B WAS USED DURING THE REBOUND PHASE TO AVOID NEW INFECTION EVENTS. RESULTS: BOTH SIRNA AND PEG-IFNALPHA STRONGLY REDUCED ALL HBV MARKERS, INCLUDING HBX LEVELS, THUS ENABLING THE REAPPEARANCE OF SMC5/6 IN HEPATOCYTES THAT ACHIEVED HBV-RNA NEGATIVISATION AND SMC5/6 ASSOCIATION WITH THE CCCDNA. ONLY IFN REDUCED CCCDNA LOADS AND ENHANCED IFN-STIMULATED GENES. HOWEVER, THE ANTIVIRAL EFFECTS DID NOT PERSIST OFF TREATMENT AND SMC5/6 WAS AGAIN DEGRADED. REMARKABLY, THE BLOCKADE OF VIRAL ENTRY THAT STARTED AT THE END OF TREATMENT HINDERED RENEWED DEGRADATION OF SMC5/6. CONCLUSION: THESE RESULTS REVEAL THAT THERAPEUTICS ABROGATING ALL HBV TRANSCRIPTS INCLUDING HBX PROMOTE EPIGENETIC SUPPRESSION OF THE HBV MINICHROMOSOME, WHEREAS STRATEGIES PROTECTING THE HUMAN HEPATOCYTES FROM REINFECTION ARE NEEDED TO MAINTAIN CCCDNA SILENCING.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
17 4225  33 METHYLATION DEGREE OF METALLOPROTEINASE INHIBITOR RECK GENE: LINKS TO RECK PROTEIN LEVEL AND HEPATOCELLULAR CARCINOMA IN CHRONIC HCV INFECTION PATIENTS. THE RECK GENE, A TUMOR SUPPRESSOR GENE, INHIBITS ANGIOGENESIS, INVASION, AND TUMOR METASTASIS. EPIGENETIC REGULATION OF THE RECK GENE CONSTITUTES A POTENT APPROACH TO THE MOLECULAR BASIS OF LIVER MALIGNANCY. THIS STUDY AIMS TO EVALUATE THE PROMOTER METHYLATION STATUS OF THE RECK GENE AND ITS SERUM LEVEL IN PATIENTS WITH HEPATITIS C VIRUS (HCV)-RELATED HEPATOCELLULAR CARCINOMA (HCC) AND THE POTENTIAL ASSOCIATION OF RECK GENE METHYLATION WITH CLINICAL CRITERIA OF HCC. ONE HUNDRED AND FIFTY-FIVE SUBJECTS WERE INCLUDED (HEALTHY CONTROL [55], CHRONIC HCV PATIENTS [55], HCV-RELATED HCC PATIENTS [45]). THE METHYLATION STATUS OF THE RECK GENE PROMOTER AND SERUM RECK LEVEL WERE INVESTIGATED BY METHYLATION-SPECIFIC PCR AND ENZYME-LINKED IMMUNOSORBENT ASSAY TECHNIQUES, RESPECTIVELY. RECK GENE PROMOTER HYPERMETHYLATION WAS RECORDED IN 46.7% OF HCC PATIENTS, AND 10.9% OF HCV PATIENTS, BUT NOT IN CONTROL SUBJECTS (0%). IT WAS RELATED TO RECK PROTEIN LEVEL, VARICES, EDEMA, ASCITES, LYMPH NODE METASTASIS, VASCULAR INVASION, AND THE LARGEST DIAMETER OF FOCAL LESIONS. MEANWHILE, IT WAS NOT ASSOCIATED WITH FOCAL LESION NUMBER NOR DISTANT METASTASIS OF HCC. IN CONCLUSION, RECK GENE PROMOTER HYPERMETHYLATION IS LINKED TO HCV GENOTYPE-4-RELATED HCC. MOREOVER, DIFFERENT DEGREES OF RECK GENE PROMOTER METHYLATION ARE ASSOCIATED WITH SERUM RECK LEVEL, LYMPH NODE METASTASIS, AND VASCULAR INVASION, WHICH COULD PROVE ITS PATHOGENIC ROLE IN HEPATOCARCINOGENESIS IN CHRONIC HCV-INFECTED PATIENTS.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
18 4443  39 MOLECULAR INSIGHTS INTO SPINDLIN1-HBX INTERPLAY AND ITS IMPACT ON HBV TRANSCRIPTION FROM CCCDNA MINICHROMOSOME. MOLECULAR INTERPLAY BETWEEN HOST EPIGENETIC FACTORS AND VIRAL PROTEINS CONSTITUTES AN INTRIGUING MECHANISM FOR SUSTAINING HEPATITIS B VIRUS (HBV) LIFE CYCLE AND ITS CHRONIC INFECTION. HBV ENCODES A REGULATORY PROTEIN, HBX, WHICH ACTIVATES TRANSCRIPTION AND REPLICATION OF HBV GENOME ORGANIZED AS COVALENTLY CLOSED CIRCULAR (CCC) DNA MINICHROMOSOME. HERE WE ILLUSTRATE HOW HBX ACCOMPLISHES ITS TASK BY HIJACKING SPINDLIN1, AN EPIGENETIC READER COMPRISING THREE CONSECUTIVE TUDOR DOMAINS. OUR BIOCHEMICAL AND STRUCTURAL STUDIES HAVE REVEALED THAT THE HIGHLY CONSERVED N-TERMINAL 2-21 SEGMENT OF HBX (HBX(2-21)) ASSOCIATES INTIMATELY WITH TUDOR 3 OF SPINDLIN1, ENHANCING HISTONE H3 "K4ME3-K9ME3" READOUT BY TUDORS 2 AND 1. FUNCTIONALLY, SPINDLIN1-HBX ENGAGEMENT PROMOTES GENE EXPRESSION FROM THE CHROMATINIZED CCCDNA, ACCOMPANIED BY AN EPIGENETIC SWITCH FROM AN H3K9ME3-ENRICHED REPRESSIVE STATE TO AN H3K4ME3-MARKED ACTIVE STATE, AS WELL AS A CONFORMATIONAL SWITCH OF HBX THAT MAY OCCUR IN COORDINATION WITH OTHER HBX-BINDING FACTORS, SUCH AS DDB1. DESPITE A PROPOSED TRANSREPRESSION ACTIVITY OF HBX(2-21), OUR STUDY REVEALS A KEY ROLE OF SPINDLIN1 IN DEREPRESSING THIS CONSERVED MOTIF, THEREBY PROMOTING HBV TRANSCRIPTION FROM ITS CHROMATINIZED GENOME.	2023	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
19 1966  32 EPIGENETIC ALTERATION OF PRKCDBP IN COLORECTAL CANCERS AND ITS IMPLICATION IN TUMOR CELL RESISTANCE TO TNFALPHA-INDUCED APOPTOSIS. PURPOSE: PRKCDBP IS A PUTATIVE TUMOR SUPPRESSOR IN WHICH ALTERATION HAS BEEN OBSERVED IN SEVERAL HUMAN CANCERS. WE INVESTIGATED EXPRESSION AND FUNCTION OF PRKCDBP IN COLORECTAL CELLS AND TISSUES TO EXPLORE ITS CANDIDACY AS A SUPPRESSOR IN COLORECTAL TUMORIGENESIS. EXPERIMENTAL DESIGN: EXPRESSION AND METHYLATION STATUS OF PRKCDBP AND ITS EFFECT ON TUMOR GROWTH WERE EVALUATED. TRANSCRIPTIONAL REGULATION BY NF-KAPPAB SIGNALING WAS DEFINED BY LUCIFERASE REPORTER AND CHROMATIN IMMUNOPRECIPITATION ASSAYS. RESULTS: PRKCDBP EXPRESSION WAS HARDLY DETECTABLE IN 29 OF 80 (36%) PRIMARY TUMORS AND 11 OF 19 (58%) CELL LINES, AND ITS ALTERATION CORRELATED WITH TUMOR STAGE AND GRADE. PROMOTER HYPERMETHYLATION WAS COMMONLY FOUND IN CANCERS. PRKCDBP EXPRESSION INDUCED THE G(1) CELL-CYCLE ARREST AND INCREASED CELLULAR SENSITIVITY TO VARIOUS APOPTOTIC STRESSES. PRKCDBP WAS INDUCED BY TNFALPHA, AND ITS LEVEL CORRELATED WITH TUMOR CELL SENSITIVITY TO TNFALPHA-INDUCED APOPTOSIS. PRKCDBP INDUCTION BY TNFALPHA WAS DISRUPTED BY BLOCKING NF-KAPPAB SIGNALING WHILE IT WAS ENHANCED BY RELA TRANSFECTION. THE PRKCDBP PROMOTER ACTIVITY WAS INCREASED IN RESPONSE TO TNFALPHA, AND THIS RESPONSE WAS ABOLISHED BY DISRUPTION OF A KAPPAB SITE IN THE PROMOTER. PRKCDBP DELAYED THE FORMATION AND GROWTH OF XENOGRAFT TUMORS AND IMPROVED TUMOR RESPONSE TO TNFALPHA-INDUCED APOPTOSIS. CONCLUSIONS: PRKCDBP IS A PROAPOPTOTIC TUMOR SUPPRESSOR WHICH IS COMMONLY ALTERED IN COLORECTAL CANCER BY PROMOTER HYPERMETHYLATION, AND ITS GENE TRANSCRIPTION IS DIRECTLY ACTIVATED BY NF-KAPPAB IN RESPONSE TO TNFALPHA. THIS SUGGESTS THAT PRKCDBP INACTIVATION MAY CONTRIBUTE TO TUMOR PROGRESSION BY REDUCING CELLULAR SENSITIVITY TO TNFALPHA AND OTHER STRESSES, PARTICULARLY UNDER CHRONIC INFLAMMATORY MICROENVIRONMENT.	2011	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
20 4233  36 METHYLATION OF SEPTIN9 MEDIATED BY DNMT3A ENHANCES HEPATIC STELLATE CELLS ACTIVATION AND LIVER FIBROGENESIS. LIVER FIBROSIS, RESULTING FROM CHRONIC AND PERSISTENT INJURY TO THE LIVER, IS A WORLDWIDE HEALTH PROBLEM. ADVANCED LIVER FIBROSIS RESULTS IN CIRRHOSIS, LIVER FAILURE AND EVEN HEPATOCELLULAR CANCER (HCC), OFTEN EVENTUALLY REQUIRING LIVER TRANSPLANTATION, POSES A HUGE HEALTH BURDEN ON THE GLOBAL COMMUNITY. HOWEVER, THE SPECIFIC PATHOGENESIS OF LIVER FIBROSIS REMAINS NOT FULLY UNDERSTOOD. NUMEROUS BASIC AND CLINICAL STUDIES HAVE PROVIDED EVIDENCE THAT EPIGENETIC MODIFICATIONS, ESPECIALLY DNA METHYLATION, MIGHT CONTRIBUTE TO THE ACTIVATION OF HEPATIC STELLATE CELLS (HSCS), THE PIVOTAL CELL TYPE RESPONSIBLE FOR THE FIBROUS SCAR IN LIVER. HERE, REDUCED REPRESENTATION BISULFITE SEQUENCING (RRBS) AND BISULFITE PYROSEQUENCING PCR (BSP) ANALYSIS IDENTIFIED HYPERMETHYLATION STATUS OF SEPTIN9 (SEPT9) GENE IN LIVER FIBROGENESIS. SEPT9 PROTEIN WAS DRAMATICALLY DECREASED IN LIVERS OF CCL4-TREATED MICE AND IMMORTALIZED HSC-T6 CELLS EXPOSED TO TGF-BETA1. NEVERTHELESS, THE SUPPRESSION OF SEPT9 COULD BE BLOCKED BY DNMT3A-SIRNA AND DNA METHYLTRANSFERASE INHIBITOR, 5-AZA-2'-DEOXYCYTIDINE (5-AZADC). OVEREXPRESSED SEPT9 ATTENUATED TGF-BETA1-INDUCED EXPRESSION OF MYOFIBROBLAST MARKERS ALPHA-SMA AND COL1A1, ACCOMPANIED BY UP-REGULATION OF CELL APOPTOSIS-RELATED PROTEINS. CONVERSELY, RNAI-MEDIATED SILENCING OF SEPT9 ENHANCED ACCUMULATION OF EXTRACELLULAR MATRIX. THESE OBSERVATIONS SUGGESTED THAT SEPT9 CONTRIBUTED TO ALLEVIATE LIVER FIBROSIS MIGHT PARTIALLY THROUGH PROMOTING ACTIVATED HSCS APOPTOSIS AND THIS ANTI-FIBROGENESIS EFFECT MIGHT BE BLOCKED BY DNMT-3A MEDIATED METHYLATION OF SEPT9. THEREFORE, PHARMACOLOGICAL AGENTS THAT INHIBIT SEPT9 METHYLATION AND INCREASE ITS EXPRESSION COULD BE CONSIDERED AS VALUABLE TREATMENTS FOR LIVER FIBROSIS.	2017