1  3185 102 HBC BINDS TO THE CPG ISLANDS OF HBV CCCDNA AND PROMOTES AN EPIGENETIC PERMISSIVE STATE. HBV COVALENTLY CLOSED CIRCULAR DNA (CCCDNA) IS THE TEMPLATE FOR THE TRANSCRIPTION OF HBV. HBV CORE PROTEIN (HBC) IS A MAIN COMPONENT OF THE HBV CCCDNA MINICHROMOSOME. HOWEVER, THE FUNCTION OF HBC IN CCCDNA IS NOT FULLY UNDERSTOOD. IN LIGHT OF RECENT FINDINGS THAT HBV CCCDNA MAY BE REGULATED EPIGENETICALLY, WE ANALYZED THE BINDING OF HBC TO CCCDNA AND THE IMPACT OF HBC ON CCCDNA EPIGENETIC PROFILE IN THE LIVER BIOPSY SAMPLES OF 22 PATIENTS WITH CHRONIC HEPATITIS B (CHB). WE FOUND THAT HBC BINDING TO HBV CCCDNA OCCURRED PREFERENTIALLY AT CPG ISLAND 2, AN IMPORTANT REGION FOR THE REGULATION OF HBV TRANSCRIPTION. FURTHERMORE, THE RELATIVE ABUNDANCES OF HBC BINDING TO CPG ISLAND 2 WERE POSITIVELY CORRELATED WITH THE RATIOS OF RELAXED CIRCULAR DNA TO CCCDNA AND THE LEVELS OF SERUM HBV DNA IN THOSE PATIENTS. INTERESTINGLY, THE RELATIVE ABUNDANCES OF HBC BINDING TO CPG ISLAND 2 WERE ASSOCIATED WITH THE BINDING OF CREB BINDING PROTEIN (CBP) AND WITH HYPOMETHYLATION IN CPG ISLAND 2 OF HBV CCCDNA MINICHROMOSOMES. HOWEVER, RELATIVELY HIGHER AMOUNTS OF HBC BINDING TO CPG ISLAND 2 OF CCCDNA WERE ACCOMPANIED BY LOWER AMOUNTS OF HDAC1 BINDING. MULTIVARIATE ANALYSIS REVEALED THAT THE ABUNDANCES OF HBC BINDING TO CPG ISLAND 2 OF CCCDNA AND POSITIVE HBEAG WERE INDEPENDENT FACTORS ASSOCIATED WITH THE REPLICATION OF HBV (P = 0.001 FOR BOTH). APPARENTLY, HBC IS A POSITIVE REGULATOR OF HBV TRANSCRIPTION AND REPLICATION, MAINTAINING THE PERMISSIVE EPIGENETIC STATE IN THE CRITICAL REGION OF THE HBV CCCDNA MINICHROMOSOMES.	2011	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
2  1190  29 CORRELATION BETWEEN HEPATIC HUMAN MALES ABSENT ON THE FIRST (HMOF) AND VIRAL PERSISTENCE IN CHRONIC HEPATITIS B PATIENTS. BACKGROUND: CHRONIC HEPATITIS B (CHB) REMAINS A GLOBAL HEALTH DILEMMA WITH HIGH MORBIDITY AND MORTALITY. HUMAN MALES ABSENT ON THE FIRST (HMOF) (A HISTONE ACETYLTRANSFERASE) IS RESPONSIBLE FOR DNA DAMAGE REPAIR, TUMORIGENESIS AND CELL CYCLE REGULATION. PERSISTENCE OF HBV DNA CONTRIBUTES TO CIRRHOSIS AND HEPATOCELLULAR CARCINOMA (HCC) IN CHB PATIENTS. HISTONE ACETYLTRANSFERASE ENHANCES HBV REPLICATION, HOWEVER THE PRECISE UNDERLYING MECHANISM OF HMOF IN HBV REPLICATION IN CHB PATIENTS REMAINS TO BE EXPLORED. THIS STUDY AIMS TO INVESTIGATE THE CORRELATION BETWEEN HEPATIC HMOF AND HBV DNA REPLICATION IN CHB PATIENTS, AND MAY PROVIDE NEW INSIGHTS TOWARDS THE TREATMENT OF CHB PATIENTS. METHODS: HMOF IN LIVER BIOPSY (CHB, N = 33 HBEAG(+); N = 20 HBEAG(-), AND THREE HEALTHY CONTROLS) WAS DETERMINED, USING IMMUNOHISTOCHEMISTRY, QPCR AND WESTERN BLOT. THE CORRELATION BETWEEN HMOF AND HBSAG, AS WELL AS, HBEAG WERE DETERMINED. RESULTS: A POSITIVE CORRELATION BETWEEN HMOF AND HBV DNA IN OVERALL CHB PATIENTS WAS OBSERVED. A DISTINCT POSITIVE CORRELATION BETWEEN HMOF AND HBSAG AND/OR HBEAG IN HBEAG(+) CHB PATIENTS WAS ALSO DETECTED, HOWEVER NOT OBSERVED BETWEEN HMOF AND HBSAG IN HBEAG(-) CHB PATIENTS. NO CORRELATION WAS OBSERVED BETWEEN HMOF AND HEPATIC INFLAMMATION SEVERITY AND FIBROTIC STAGE IN CHB PATIENTS. CONCLUSIONS: HEPATIC HMOF MIGHT CONTRIBUTE TO HOST HBV CLEARANCE IN CHB PATIENTS AND POSSIBLE PATHOGENESIS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
3  3246  27 HEPATITIS B VIRUS (HBV) INDUCES THE EXPRESSION OF INTERLEUKIN-8 THAT IN TURN REDUCES HBV SENSITIVITY TO INTERFERON-ALPHA. HIGH LEVELS OF SERUM INTERLEUKIN-8 (IL-8) HAVE BEEN DETECTED IN CHRONIC HEPATITIS B (CHB) PATIENTS DURING EPISODES OF HEPATITIS FLARES. WE INVESTIGATED WHETHER HEPATITIS B VIRUS (HBV) MAY DIRECTLY INDUCE IL-8 PRODUCTION AND WHETHER IL-8 MAY ANTAGONIZE INTERFERON-ALPHA (IFN-ALPHA) ANTIVIRAL ACTIVITY AGAINST HBV. WE SHOWED THAT CHB PATIENTS HAD SIGNIFICANTLY HIGHER IL-8 LEVELS BOTH IN SERUM AND IN LIVER TISSUE THAN CONTROLS. IN HBV-REPLICATING HEPG2 CELLS, IL-8 TRANSCRIPTION WAS SIGNIFICANTLY ACTIVATED. AP-1, C/EBP AND NF-KB TRANSCRIPTION FACTORS WERE CONCURRENTLY NECESSARY FOR MAXIMUM IL-8 INDUCTION. MOREOVER, HBX VIRAL PROTEIN WAS RECRUITED ONTO THE IL-8 PROMOTER AND THIS WAS PARALLELED BY IL8-BOUND HISTONE HYPERACETYLATION AND BY ACTIVE RECRUITMENT OF TRANSCRIPTIONAL COACTIVATORS. INHIBITION OF IL-8 INCREASES THE ANTIVIRAL ACTIVITY OF IFN-ALPHA AGAINST HBV. OUR RESULTS INDICATE THAT HBV ACTIVATES IL-8 GENE EXPRESSION BY TARGETING THE EPIGENETIC REGULATION OF THE IL-8 PROMOTER AND THAT IL-8 MAY CONTRIBUTE TO REDUCE HBV SENSITIVITY TO IFN-ALPHA.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
4  4239  31 METHYLATION PROFILE OF HEPATITIS B VIRUS IS NOT INFLUENCED BY INTERFERON ALPHA IN HUMAN LIVER CANCER CELLS. INTERFERON (IFN) ALPHA IS USED FOR THE TREATMENT OF CHRONIC HEPATITIS B VIRUS (HBV) INFECTION, BUT THE MOLECULAR MECHANISMS UNDERLYING ITS ANTIVIRAL EFFECT HAVE NOT BEEN FULLY ELUCIDATED. EPIGENETIC MODIFICATIONS REGULATE THE TRANSCRIPTIONAL ACTIVITY OF COVALENTLY CLOSED CIRCULAR DNA (CCCDNA) IN CELLS WITH CHRONIC HBV INFECTION. IFN?ALPHA HAS BEEN SHOWN TO MODIFY CCCDNA?BOUND HISTONES, BUT IT IS NOT KNOWN WHETHER THE ANTI?HBV EFFECT OF IFN?ALPHA INVOLVES METHYLATION OF CCCDNA. THE PRESENT STUDY AIMED TO DETERMINE WHETHER IFN?ALPHA INDUCED METHYLATION OF HBV CCCDNA IN A CELL?BASED MODEL IN WHICH HEPG2 CELLS WERE DIRECTLY INFECTED WITH WILD?TYPE HBV VIRIONS. METHYLATION STATUS OF HBV CCCDNA WAS ASSESSED USING GLOBAL DNA METHYLATION ELISA ASSAY, METHYLATION?SPECIFIC PCR AND BISULFITE SEQUENCING. IFN?ALPHA SUPPRESSED HBV DNA AND RNA TRANSCRIPTS, BUT METHYLATION PROFILES WERE SIMILAR BETWEEN THE CONTROL AND IFN?ALPHA TREATED GROUPS. CHROMATIN IMMUNOPRECIPITATION RESULTS REVEALED BINDING OF DNA METHYLTRANSFERASES (DNMT) 3A AND DNMT3B TO HBV CCCDNA AND TREATMENT WITH IFN?ALPHA SUPPRESSED THE RECRUITMENT OF DNMT3B TO CCCDNA. TAKEN TOGETHER, THESE RESULTS SUGGEST THAT IFN?ALPHA DOES NOT INDUCE METHYLATION OF HBV CCCDNA. THEREFORE, IT WAS CONCLUDED THAT METHYLATION IS UNLIKELY TO CONTRIBUTE TO THE ANTI?HBV EFFECT OF IFN?ALPHA IN HEPG2 CELLS, AND THAT ALTERNATIVE MECHANISMS NEED TO BE SOUGHT TO ENHANCE CCCDNA METHYLATION AS A NOVEL THERAPY AGAINST HBV.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
5  1393  32 DIAGNOSTIC VALUE OF THE HYPOMETHYLATION OF THE WISP1 PROMOTER IN PATIENTS WITH HEPATOCELLULAR CARCINOMA ASSOCIATED WITH HEPATITIS B VIRUS. WNT1-INDUCIBLE SIGNALING PATHWAY PROTEIN 1 (WISP1) REGULATES CELL PROLIFERATION, DIFFERENTIATION, ADHESION, MIGRATION AND SURVIVAL. ABNORMAL WISP1 EXPRESSION IS ASSOCIATED WITH THE CARCINOGENESIS OF HEPATOCELLULAR CARCINOMA (HCC). ABERRANT DNA METHYLATION IS ONE OF THE MAJOR EPIGENETIC ALTERATIONS IN HCC. HOWEVER, THE METHYLATION STATUS OF THE WISP1 PROMOTER IS STILL UNCLEAR. WE THEREFORE AIMED TO DETERMINE THE METHYLATION STATUS OF THE WISP1 PROMOTER AND EVALUATE ITS CLINICAL VALUE IN HCC. THE STUDY ENROLLED 251 PARTICIPANTS, INCLUDING 123 PARTICIPANTS WITH HCC, 90 PARTICIPANTS WITH CHRONIC HEPATITIS B (CHB) AND 38 HEALTHY CONTROLS (HCS). WISP1 METHYLATION STATUS, MRNA LEVELS AND PLASMA SOLUBLE WISP1 WERE DETECTED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP), QUANTITATIVE REAL-TIME PCR (RT-QPCR) AND ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA), RESPECTIVELY. WE FOUND THAT THE METHYLATION FREQUENCY OF WISP1 IN PATIENTS WITH HCC WAS SIGNIFICANTLY LOWER THAN THAT IN PATIENTS WITH CHB AND HCS, WHILE THE RELATIVE EXPRESSION LEVELS OF WISP1 MRNA WERE MARKEDLY HIGHER IN PATIENTS WITH HCC THAN IN PATIENTS WITH CHB AND HCS. FURTHERMORE, THE PLASMA SOLUBLE WISP1 IN PATIENTS WITH HCC WAS OBVIOUSLY LOWER THAN IN THAT IN PATIENTS WITH CHB AND HCS. ALPHA-FETOPROTEIN (AFP) IS A WIDELY RECOGNIZED BIOMARKER TO DIAGNOSE HCC WHICH LACKS ENOUGH SENSITIVITY AND SPECIFICITY. WISP1 PROMOTER METHYLATION STATUS COMBINED WITH AFP SIGNIFICANTLY IMPROVED THE DIAGNOSTIC ABILITY IN DISCRIMINATING HCC FROM CHB COMPARED WITH AFP OR WISP1 METHYLATION STATUS ALONE. IN CONCLUSION, HYPOMETHYLATION OF THE WISP1 GENE PROMOTER MAY SERVE AS A NONINVASIVE BIOMARKER FOR DETECTING HBV-ASSOCIATED HCC.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
6  4903  22 P16 PROMOTER HYPERMETHYLATION IN HUMAN HEPATOCELLULAR CARCINOMA WITH OR WITHOUT HEPATITIS VIRUS INFECTION. BACKGROUND: EPIGENETIC ALTERATION THROUGH METHYLATION IS ONE OF THE MOST IMPORTANT STEPS IN CARCINOGENESIS. HOWEVER, THE RELATION BETWEEN HEPATITIS VIRUS INFECTION AND EPIGENETIC ALTERATIONS IS POORLY UNDERSTOOD. METHODS: SIXTEEN PATIENTS WITHOUT HEPATITIS B VIRUS (HBV) AND HEPATITIS C VIRUS (HCV) AND 35 PATIENTS WITH HBV OR HCV WHO UNDERWENT LIVER RESECTION FOR HEPATOCELLULAR CARCINOMA (HCC) WERE STUDIED. MUTATION OF P53 WAS DETECTED BY DIRECT SEQUENCING. METHYLATION STATUS OF P16 WAS EVALUATED IN TUMOR AND NONCANCEROUS LIVER TISSUES BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. RESULTS: IN HCC WITHOUT HBV AND HCV, P53 MUTATIONS WERE DETECTED IN 5 (31%) OF 16 HCCS. METHYLATION OF P16 PROMOTER WAS DETECTED IN 2 (25%) OF 8 MODERATELY DIFFERENTIATED HCCS, 6 (75%) OF 8 POORLY DIFFERENTIATED HCCS, AND NONE OF 16 NONCANCEROUS TISSUE SPECIMENS. IN HCC WITH HBV OR HCV, P53 MUTATIONS WERE DETECTED IN 8 (23%) OF 35 HCCS. METHYLATION OF P16 PROMOTER WAS DETECTED IN 2 (100%) OF 2 WELL-DIFFERENTIATED HCCS, 13 (76%) OF 17 MODERATELY DIFFERENTIATED HCCS, 12 (75%) OF 16 POORLY DIFFERENTIATED HCCS, AND 9 (26%) OF 35 NONCANCEROUS LIVER TISSUE SPECIMENS. CONCLUSIONS: OUR RESULTS SUGGEST THAT HEPATITIS VIRUSES MIGHT INDUCE METHYLATION OF P16 PROMOTER IN LIVER WITH CHRONIC INFLAMMATION, BEFORE APPEARANCE OF HCC.	2004	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
7   412  29 ANALYSIS OF LONG NON-CODING RNA (LNCRNA) EXPRESSION IN HEPATITIS B PATIENTS. LONG NON-CODING RNAS (LNCRNAS) HAVE BEEN IMPLICATED IN NUMEROUS BIOLOGICAL PROCESSES, INCLUDING EPIGENETIC REGULATION, CELL-CYCLE CONTROL, AND TRANSCRIPTIONAL/TRANSLATIONAL REGULATION OF GENE EXPRESSION. DIFFERENTIAL EXPRESSION OF LNCRNAS AND DISRUPTION OF THE REGULATORY PROCESSES ARE RECOGNIZED AS CRITICAL STEPS IN CANCER DEVELOPMENT. THE ROLE OF LNCRNAS IN HEPATITIS B VIRUS (HBV) INFECTION IS NOT WELL UNDERSTOOD. HERE WE ANALYZED THE EXPRESSION OF 135 LNCRNAS IN PLASMA SAMPLES OF 82 HBV PATIENTS (CLASSIFIED AS CHRONIC PATIENTS, INACTIVE CARRIERS, OR RESOLVED PATIENTS) AT DIAGNOSIS AND AT 12 MONTHS OF TREATMENT IN RELATION TO CONTROL GROUP (81 HEALTHY VOLUNTEERS). WE ALSO INVESTIGATED THE EFFECT OF SMALL INTERFERING RNA (SIRNA)-MEDIATED SILENCING OF LINCRNA-SFMBT2 ON HBV-POSITIVE HUMAN LIVER CANCER CELL LINE. LNCRNA EXPRESSION WAS ANALYZED BY QUANTITATIVE REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION (QRT-PCR). CHEMICALLY SYNTHESIZED SIRNAS WERE TRANSFECTED INTO THE CELL LINES USING LIPOFECTAMINE 2000 REAGENT (THERMO FISHER SCIENTIFIC). HBV DNA AND HBSAG AND HBEAG WERE DETECTED IN TRANSFECTED CULTURES BY REAL-TIME PCR AND ELISA, RESPECTIVELY, USING COMMERCIAL KITS. WE OBSERVED CHANGES IN LNCRNA EXPRESSION IN ALL THREE HBV GROUPS, COMPARED TO CONTROL GROUP. MOST NOTABLY, THE EXPRESSION OF ANTI-NOS2A, LINCRNA-SFMBT2, AND ZFHX2AS WAS SIGNIFICANTLY INCREASED AND EXPRESSION OF Y5 LNCRNA WAS DECREASED IN CHRONIC HBV PATIENTS. A DECREASED Y5 EXPRESSION AND INCREASED LINCRNA-SFMBT2 EXPRESSION WERE OBSERVED IN INACTIVE HBSAG CARRIERS. THE EXPRESSION OF HOTTIP, MEG9, AND PCAT-32 WAS INCREASED IN RESOLVED HBV PATIENTS, AND NO SIGNIFICANT CHANGE IN THE EXPRESSION OF Y5 WAS OBSERVED, COMPARED TO CONTROL GROUP. SIRNA-MEDIATED INHIBITION OF LINCRNA-SFMBT2 DECREASED THE LEVEL OF HBV DNA IN HUMAN LIVER CANCER CELLS. FURTHER RESEARCH IS NEEDED TO CONFIRM THE PROGNOSTIC AS WELL AS THERAPEUTIC ROLE OF THESE LNCRNAS IN HBV PATIENTS.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
8  3186  32 HBV COVALENTLY CLOSED CIRCULAR DNA MINICHROMOSOMES IN DISTINCT EPIGENETIC TRANSCRIPTIONAL STATES DIFFER IN THEIR VULNERABILITY TO DAMAGE. BACKGROUND AND AIMS: HBV COVALENTLY CLOSED CIRCULAR DNA (CCCDNA) IS A MAJOR OBSTACLE FOR A CURE OF CHRONIC HEPATITIS B. ACCUMULATING EVIDENCE SUGGESTS THAT EPIGENETIC MODIFICATIONS REGULATE THE TRANSCRIPTIONAL ACTIVITY OF CCCDNA MINICHROMOSOMES. HOWEVER, IT REMAINS UNCLEAR HOW THE EPIGENETIC STATE OF CCCDNA AFFECTS ITS STABILITY. APPROACHES AND RESULTS: BY USING HBV INFECTION CELL MODELS AND IN VITRO AND IN VIVO RECOMBINANT CCCDNA (RCCCDNA) AND HBVCIRCLE MODELS, THE REDUCTION RATE OF HBV CCCDNA AND THE EFFICACY OF APOLIPOPROTEIN B MRNA EDITING ENZYME CATALYTIC SUBUNIT 3A (APOBEC3A)-MEDIATED AND CRISPR/CRISPR-ASSOCIATED 9 (CAS9)-MEDIATED CCCDNA TARGETING WERE COMPARED BETWEEN CCCDNAS WITH DISTINCT TRANSCRIPTIONAL ACTIVITIES. INTERFERON-ALPHA TREATMENT AND HEPATITIS B X PROTEIN (HBX) DELETION WERE APPLIED AS TWO STRATEGIES FOR CCCDNA REPRESSION. CHROMATIN IMMUNOPRECIPITATION AND MICROCOCCAL NUCLEASE ASSAYS WERE PERFORMED TO DETERMINE THE EPIGENETIC PATTERN OF CCCDNA. HBV CCCDNA LEVELS REMAINED STABLE IN NONDIVIDING HEPATOCYTES; HOWEVER, THEY WERE SIGNIFICANTLY REDUCED DURING CELL DIVISION, AND THE REDUCTION RATE WAS SIMILAR BETWEEN CCCDNAS IN TRANSCRIPTIONALLY ACTIVE AND TRANSCRIPTIONALLY REPRESSED STATES. STRIKINGLY, HBV RCCCDNA WITHOUT HBX EXPRESSION EXHIBITED A SIGNIFICANTLY LONGER PERSISTENCE IN MICE. THE CCCDNA WITH LOW TRANSCRIPTIONAL ACTIVITY EXHIBITED AN EPIGENETICALLY INACTIVE PATTERN AND WAS MORE DIFFICULT TO ACCESS BY APOBEC3A AND ENGINEERED CRISPR-CAS9. THE EPIGENETIC REGULATOR ACTIVATING CCCDNA INCREASED ITS VULNERABILITY TO APOBEC3A. CONCLUSIONS: HBV CCCDNA MINICHROMOSOMES IN DISTINCT EPIGENETIC TRANSCRIPTIONAL STATES SHOWED A SIMILAR REDUCTION RATE DURING CELL DIVISION BUT SIGNIFICANTLY DIFFERED IN THEIR ACCESSIBILITY AND VULNERABILITY TO TARGETED NUCLEASES AND ANTIVIRAL AGENTS. EPIGENETIC SENSITIZATION OF CCCDNA MAKES IT MORE SUSCEPTIBLE TO DAMAGE AND MAY POTENTIALLY CONTRIBUTE TO AN HBV CURE.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                
9  3794  37 INTERLEUKIN-33 MEDIATES BOTH IMMUNE-RELATED AND NON-IMMUNE-RELATED INHIBITORY EFFECTS AGAINST HEPATITIS B VIRUS. CHRONIC INFECTION BY HEPATITIS B VIRUS (HBV) IS ASSOCIATED WITH HIGH RISKS OF LIVER FIBROSIS, CIRRHOSIS AND HEPATOCELLULAR CARCINOMA. HBV COVALENTLY CLOSED CIRCULAR DNA (CCCDNA) IN THE NUCLEUS OF INFECTED HEPATOCYTE SERVES AS TRANSCRIPTION TEMPLATE. NEITHER NATURAL RESOLUTION OF ACUTE INFECTION NOR CURRENT TREATMENT OPTIONS FOR CHRONIC INFECTION ARE BELIEVED TO CAUSE CCCDNA CLEARANCE. WE PREVIOUSLY SHOWED THAT INJECTION OF IL-33-EXPRESSING PLASMID FACILITATED CLEARANCE OF INTRAHEPATIC HBV DNA IN A MOUSE MODEL OF HBV PERSISTENCE. IN THIS WORK, HBV-TARGETING THERAPEUTIC EFFECTS OF IL-33 WERE FURTHER EXPLORED. MURINE IL-33 DELIVERED BY RECOMBINANT ADENO-ASSOCIATED VIRUS (AAV-MIL-33) INDUCED CLEARANCE OF BOTH SERUM HBV MARKERS AND INTRAHEPATIC HBV DNA IN TWO MOUSE MODELS OF HBV PERSISTENCE BASED ON REPLICON PLASMID AND RECOMBINANT CCCDNA (RCCCDNA) RESPECTIVELY. CLEARANCE WAS ASSOCIATED WITH SERUM ALT ELEVATIONS AND LIVER INFILTRATIONS BY CD4(+) AND CD8(+) T CELLS, INDICATING IL-33-INDUCED CELLULAR IMMUNE RESPONSES AGAINST HBV-HARBORING CELLS. ADOPTIVE TRANSFER OF SPLENOCYTES FROM AAV-MIL-33-CURED MICE WAS INDEED SUFFICIENT TO ENGENDER SIMILAR CLEARANCE IN RECIPIENT MICE. IN VITRO, INTRACELLULAR, INSTEAD OF EXTRACELLULAR, IL-33 WAS MAINLY RESPONSIBLE FOR REPRESSING VIRAL TRANSCRIPTION, PROTEIN PRODUCTION AND GENOME REPLICATION IN HUH7 CELLS TRANSFECTED WITH HBV REPLICON OR RCCCDNA. IL-33 WAS SHOWN TO BE RECRUITED ONTO RCCCDNA MINICHROMOSOME ACCOMPANIED BY LOSS OF TRANSCRIPTIONAL ACTIVATION EPIGENETIC MARKS. FINALLY, TRANSFECTION OF IL-33 INTO HBV-INFECTED HEPG2/NTCP CELLS RESULTED IN REDUCED TRANSCRIPTION, ANTIGEN EXPRESSION AND GENOME REPLICATION, SUGGESTING REPRESSION OF CANONICAL CCCDNA. THESE DATA DEMONSTRATED DIVERSE INHIBITORY EFFECTS ON HBV AND HBV-INFECTED CELLS MEDIATED BY IL-33, AND SUGGEST IL-33 AS AN INTERESTING THERAPEUTIC CANDIDATE.	2022	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
10  5715  33 SIRT3 RESTRICTS HEPATITIS B VIRUS TRANSCRIPTION AND REPLICATION THROUGH EPIGENETIC REGULATION OF COVALENTLY CLOSED CIRCULAR DNA INVOLVING SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 1 AND SET DOMAIN CONTAINING 1A HISTONE METHYLTRANSFERASES. HEPATITIS B VIRUS (HBV) INFECTION REMAINS A MAJOR HEALTH PROBLEM WORLDWIDE. MAINTENANCE OF THE COVALENTLY CLOSED CIRCULAR DNA (CCCDNA), WHICH SERVES AS A TEMPLATE FOR HBV RNA TRANSCRIPTION, IS RESPONSIBLE FOR THE FAILURE OF ERADICATING CHRONIC HBV DURING CURRENT ANTIVIRAL THERAPY. CCCDNA IS ASSEMBLED WITH CELLULAR HISTONE PROTEINS INTO CHROMATIN, BUT LITTLE IS KNOWN ABOUT THE REGULATION OF HBV CHROMATIN BY HISTONE POSTTRANSLATIONAL MODIFICATIONS. IN THIS STUDY, WE IDENTIFIED SILENT MATING TYPE INFORMATION REGULATION 2 HOMOLOG 3 (SIRT3) AS A HOST FACTOR RESTRICTING HBV TRANSCRIPTION AND REPLICATION BY SCREENING SEVEN MEMBERS OF THE SIRTUIN FAMILY, WHICH IS THE CLASS III HISTONE DEACETYLASE. ECTOPIC SIRT3 EXPRESSION SIGNIFICANTLY REDUCED TOTAL HBV RNAS, 3.5-KB RNA, AS WELL AS REPLICATIVE INTERMEDIATE DNA IN HBV-INFECTED HEPG2-NA(+) /TAUROCHOLATE COTRANSPORTING POLYPEPTIDE CELLS AND PRIMARY HUMAN HEPATOCYTES. IN CONTRAST, GENE SILENCING OF SIRT3 PROMOTED HBV TRANSCRIPTION AND REPLICATION. A MECHANISTIC STUDY FOUND THAT NUCLEAR SIRT3 WAS RECRUITED TO THE HBV CCCDNA, WHERE IT DEACETYLATED HISTONE 3 LYSINE 9. IMPORTANTLY, OCCUPANCY OF SIRT3 ON CCCDNA COULD INCREASE THE RECRUITMENT OF HISTONE METHYLTRANSFERASE SUPPRESSOR OF VARIEGATION 3-9 HOMOLOG 1 TO CCCDNA AND DECREASE RECRUITMENT OF SET DOMAIN CONTAINING 1A, LEADING TO A MARKED INCREASE OF TRIMETHYL-HISTONE H3 (LYS9) AND A DECREASE OF TRIMETHYL-HISTONE H3 (LYS4) ON CCCDNA. MOREOVER, SIRT3-MEDIATED HBV CCCDNA TRANSCRIPTIONAL REPRESSION INVOLVED DECREASED BINDING OF HOST RNA POLYMERASE II AND TRANSCRIPTION FACTOR YIN YANG 1 TO CCCDNA. FINALLY, HEPATITIS B VIRAL X PROTEIN COULD RELIEVE SIRT3-MEDIATED CCCDNA TRANSCRIPTIONAL REPRESSION BY INHIBITING BOTH SIRT3 EXPRESSION AND ITS RECRUITMENT TO CCCDNA. CONCLUSION: SIRT3 IS A HOST FACTOR EPIGENETICALLY RESTRICTING HBV CCCDNA TRANSCRIPTION BY ACTING COOPERATIVELY WITH HISTONE METHYLTRANSFERASE; THESE DATA PROVIDE A RATIONALE FOR THE USE OF SIRT3 ACTIVATORS IN THE PREVENTION OR TREATMENT OF HBV INFECTION. (HEPATOLOGY 2018).	2018	
                                                                                                                                                                                                                                                   
11  3189  36 HBX RELIEVES CHROMATIN-MEDIATED TRANSCRIPTIONAL REPRESSION OF HEPATITIS B VIRAL CCCDNA INVOLVING SETDB1 HISTONE METHYLTRANSFERASE. BACKGROUND & AIMS: MAINTENANCE OF THE COVALENTLY CLOSED CIRCULAR HBV DNA (CCCDNA) THAT SERVES AS A TEMPLATE FOR HBV TRANSCRIPTION IS RESPONSIBLE FOR THE FAILURE OF ANTIVIRAL THERAPIES. WHILE STUDIES IN CHRONIC HEPATITIS PATIENTS HAVE SHOWN THAT HIGH VIREMIA CORRELATES WITH HYPERACETYLATION OF CCCDNA-ASSOCIATED HISTONES, THE MOLECULAR MECHANISMS CONTROLLING CCCDNA STABILITY AND TRANSCRIPTIONAL REGULATION ARE STILL POORLY UNDERSTOOD. THIS STUDY AIMED TO DECIPHER THE ROLE OF CHROMATIN AND CHROMATIN MODIFIER PROTEINS ON HBV TRANSCRIPTION. METHODS: WE ANALYZED THE CHROMATIN STRUCTURE OF ACTIVELY TRANSCRIBED OR SILENCED CCCDNA BY INFECTING PRIMARY HUMAN HEPATOCYTES AND DIFFERENTIATED HEPARG CELLS WITH WILD-TYPE VIRUS OR VIRUS DEFICIENT (HBVX-) FOR THE EXPRESSION OF HEPATITIS B VIRUS X PROTEIN (HBX), THAT IS REQUIRED FOR HBV EXPRESSION. RESULTS: IN THE ABSENCE OF HBX, HBV CCCDNA WAS TRANSCRIPTIONALLY SILENCED WITH THE CONCOMITANT DECREASE OF HISTONE 3 (H3) ACETYLATION AND H3K4ME3, INCREASE OF H3 DI- AND TRI-METHYLATION (H3K9ME) AND THE RECRUITMENT OF HETEROCHROMATIN PROTEIN 1 FACTORS (HP1) THAT CORRELATE WITH CONDENSED CHROMATIN. SETDB1 WAS FOUND TO BE THE MAIN HISTONE METHYLTRANSFERASE RESPONSIBLE FOR THE DEPOSITION OF H3K9ME3 AND HBV REPRESSION. FINALLY, FULL TRANSCRIPTIONAL REACTIVATION OF HBVX- UPON HBX RE-EXPRESSION CORRELATED WITH AN INCREASE OF HISTONE ACETYLATION AND H3K4ME3, AND A CONCOMITANT DECREASE OF HP1 BINDING AND OF H3K9ME3 ON THE CCCDNA. CONCLUSION: UPON HBV INFECTION, CELLULAR MECHANISMS INVOLVING SETDB1-MEDIATED H3K9ME3 AND HP1 INDUCE SILENCING OF HBV CCCDNA TRANSCRIPTION THROUGH MODULATION OF CHROMATIN STRUCTURE. HBX IS ABLE TO RELIEVE THIS REPRESSION AND ALLOW THE ESTABLISHMENT OF ACTIVE CHROMATIN.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
12  2759  33 EXPRESSION OF IL-17 AND ITS GENE PROMOTER METHYLATION STATUS ARE ASSOCIATED WITH THE PROGRESSION OF CHRONIC HEPATITIS B VIRUS INFECTION. TO EXPLORE INTERLEUKIN-17 (IL-17) AND ITS EPIGENETIC REGULATION DURING THE PROGRESSION OF CHRONIC HEPATITIS B VIRUS (HBV) INFECTION.A TOTAL OF 162 PATIENTS WITH CHRONIC HBV INFECTION, INCLUDING 75 WITH CHRONIC HEPATITIS B (CHB), 54 WITH HEPATITIS B-ASSOCIATED LIVER CIRRHOSIS AND 33 WITH HEPATITIS B-ASSOCIATED HEPATOCELLULAR CARCINOMA (HBV-HCC), WERE ENROLLED IN THIS STUDY. THIRTY HEALTHY ADULTS OF THE SAME ETHNICITY WERE ENROLLED IN THE CONTROL GROUP. WHOLE VENOUS BLOOD WAS OBTAINED FROM THE PATIENTS AND NORMAL CONTROLS (N = 30). CLINICAL AND LABORATORY PARAMETERS WERE ASSESSED, AND WE PERFORMED ENZYME-LINKED IMMUNOSORBENT ASSAY AND QUANTITATIVE REAL-TIME PCR TO MEASURE THE SERUM LEVELS AND RELATIVE MRNA EXPRESSION OF IL-17, RESPECTIVELY. IL-17 PROMOTER METHYLATION IN PERIPHERAL BLOOD MONONUCLEAR CELLS WAS ASSESSED BY METHYLATION-SPECIFIC PCR. WE ANALYZED THE SERUM AND MRNA LEVELS OF IL-17 AND IL-17 PROMOTER METHYLATION IN THE 4 GROUPS AS WELL AS THE EFFECT OF METHYLATION ON SERUM IL-17 LEVELS. CORRELATIONS BETWEEN THE IL-17 PROMOTER METHYLATION STATUS AND CLINICAL PARAMETERS WERE ANALYZED BY SPEARMAN CORRELATION ANALYSIS.COMPARED TO THE NORMAL CONTROL GROUP, THE PATIENT GROUPS EXHIBITED SIGNIFICANTLY HIGHER SERUM AND RELATIVE MRNA LEVELS OF IL-17. THE METHYLATION DISTRIBUTION AMONG THE PATIENTS WAS SIGNIFICANTLY LOWER THAN THAT AMONG THE NORMAL CONTROLS (P < .05), WITH THE HBV-HCC GROUP SHOWING THE LOWEST IL-17 GENE METHYLATION FREQUENCY. THE AVERAGE IL-17 PROMOTER CG METHYLATION LEVEL WAS NEGATIVELY CORRELATED WITH IL-17 MRNA EXPRESSION (R = -0.39, P = .03), AND NEGATIVE CORRELATIONS BETWEEN IL-17 PROMOTER METHYLATION AND PROTHROMBIN TIME ACTIVITY (R = -0.585, P = .035), ALANINE AMINOTRANSFERASE (R = -0.522, P < .01), ASPARTATE AMINOTRANSFERASE (R = -0.315, P < .05), AND THE MODEL FOR END-STAGE LIVER DISEASE SCORE (R = -0.461, P < .05) WERE OBSERVED. IL-17 SERUM LEVELS IN THE METHYLATED-PROMOTER GROUPS WERE SIGNIFICANTLY LOWER THAN THOSE IN THE UNMETHYLATED-PROMOTER GROUPS.IL-17 EXPRESSION AND PROMOTER METHYLATION WERE ASSOCIATED WITH CHRONIC HBV INFECTION PROGRESSION, ESPECIALLY IN THE HBV-HCC GROUP. THE IL-17 PROMOTER STATUS MAY HELP CLINICIANS INITIATE THE CORRECT TREATMENT STRATEGY AT THE CHB STAGE.	2019	
                                                                                                                                                
13   442  23 ANTIVIRAL THERAPIES FOR HEPATITIS B VIRUS-RELATED HEPATOCELLULAR CARCINOMA. CHRONIC HEPATITIS B VIRUS (HBV) INFECTION IS A CRITICAL RISK FACTOR FOR THE CARCINOGENESIS AND PROGRESSION OF HEPATOCELLULAR CARCINOMA (HCC). IT PROMOTES HCC DEVELOPMENT BY INDUCING LIVER FIBROGENESIS, GENETIC AND EPIGENETIC ALTERATIONS, AND THE EXPRESSION OF ACTIVE VIRAL-CODED PROTEINS. EFFECTIVE ANTIVIRAL TREATMENTS INHIBIT THE REPLICATION OF HBV, REDUCE SERUM VIRAL LOAD AND ACCELERATE HEPATITIS B E ANTIGEN SERUM CONVERSION. TIMELY INITIATION OF ANTIVIRAL TREATMENT IS NOT ONLY ESSENTIAL FOR PREVENTING THE INCIDENCE OF HCC IN CHRONIC HEPATITIS B PATIENTS, BUT ALSO IMPORTANT FOR REDUCING HBV REACTIVATION, IMPROVING LIVER FUNCTION, REDUCING OR DELAYING HCC RECURRENCE, AND PROLONGING OVERALL SURVIVAL OF HBV-RELATED HCC PATIENTS AFTER CURATIVE AND PALLIATIVE THERAPIES. THE SELECTION OF ANTIVIRAL DRUGS, MONITORING OF INDICATORS SUCH AS HBV DNA AND HEPATITIS B SURFACE ANTIGEN, AND TIMELY RESCUE TREATMENT WHEN NECESSARY, ARE ESSENTIAL IN ANTIVIRAL THERAPIES FOR HBV-RELATED HCC.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       
14  4815  34 OCCULT HEPATITIS B INFECTION AND HEPATOCELLULAR CARCINOMA: EPIDEMIOLOGY, VIROLOGY, HEPATOCARCINOGENESIS AND CLINICAL SIGNIFICANCE. OCCULT HEPATITIS B INFECTION (OBI) REFERS TO A CONDITION WHERE REPLICATION-COMPETENT HBV DNA IS PRESENT IN THE LIVER, WITH OR WITHOUT HBV DNA IN THE BLOOD, IN INDIVIDUALS WITH SERUM HBSAG NEGATIVITY ASSESSED BY CURRENTLY AVAILABLE ASSAYS. THE EPISOMAL COVALENTLY CLOSED CIRCULAR DNA (CCCDNA) IN OBI IS IN A LOW REPLICATIVE STATE. VIRAL GENE EXPRESSION IS MEDIATED BY EPIGENETIC CONTROL OF HBV TRANSCRIPTION, INCLUDING THE HBV CPG ISLAND METHYLATION PATHWAY AND POST-TRANSLATIONAL MODIFICATION OF CCCDNA-BOUND HISTONE, WITH A DIFFERENT PATTERN FROM PATIENTS WITH CHRONIC HBV INFECTION. THE PREVALENCE OF OBI VARIES TREMENDOUSLY ACROSS PATIENT POPULATIONS OWING TO NUMEROUS FACTORS, SUCH AS GEOGRAPHIC LOCATION, ASSAY CHARACTERISTICS, HOST IMMUNE RESPONSE, COINFECTION WITH OTHER VIRUSES, AND VACCINATION STATUS. APART FROM THE RISK OF VIRAL REACTIVATION UPON IMMUNOSUPPRESSION AND THE RISK OF TRANSMISSION OF HBV, OBI HAS BEEN IMPLICATED IN HEPATOCELLULAR CARCINOMA (HCC) DEVELOPMENT IN PATIENTS WITH CHRONIC HCV INFECTION, THOSE WITH CRYPTOGENIC OR KNOWN LIVER DISEASE, AND IN PATIENTS WITH HBSAG SEROCLEARANCE AFTER CHRONIC HBV INFECTION. AN INCREASING NUMBER OF PROSPECTIVE STUDIES AND META-ANALYSES HAVE REPORTED A HIGHER INCIDENCE OF HCC IN PATIENTS WITH HCV AND OBI, AS WELL AS MORE ADVANCED TUMOUR HISTOLOGICAL GRADES AND EARLIER AGE OF HCC DIAGNOSIS, COMPARED WITH PATIENTS WITHOUT OBI. THE PROPOSED PATHOGENETIC MECHANISMS OF OBI-RELATED HCC INCLUDE THE INFLUENCE OF HBV DNA INTEGRATION ON THE HEPATOCYTE CELL CYCLE, THE PRODUCTION OF PRO-ONCOGENIC PROTEINS (HBX PROTEIN AND MUTATED SURFACE PROTEINS), AND PERSISTENT LOW-GRADE NECROINFLAMMATION (CONTRIBUTING TO THE DEVELOPMENT OF FIBROSIS AND CIRRHOSIS). THERE REMAIN UNCERTAINTIES ABOUT EXACTLY HOW, AND IN WHAT ORDER, THESE MECHANISMS DRIVE THE DEVELOPMENT OF TUMOURS IN PATIENTS WITH OBI.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                
15  5226  41 PRMT5 RESTRICTS HEPATITIS B VIRUS REPLICATION THROUGH EPIGENETIC REPRESSION OF COVALENTLY CLOSED CIRCULAR DNA TRANSCRIPTION AND INTERFERENCE WITH PREGENOMIC RNA ENCAPSIDATION. CHRONIC HEPATITIS B VIRUS (HBV) INFECTION REMAINS A MAJOR HEALTH PROBLEM WORLDWIDE. THE COVALENTLY CLOSED CIRCULAR DNA (CCCDNA) MINICHROMOSOME, WHICH SERVES AS THE TEMPLATE FOR THE TRANSCRIPTION OF VIRAL RNAS, PLAYS A KEY ROLE IN VIRAL PERSISTENCE. WHILE ACCUMULATING EVIDENCE SUGGESTS THAT CCCDNA TRANSCRIPTION IS REGULATED BY EPIGENETIC MACHINERY, PARTICULARLY THE ACETYLATION OF CCCDNA-BOUND HISTONE 3 (H3) AND H4, THE POTENTIAL CONTRIBUTIONS OF HISTONE METHYLATION AND RELATED HOST FACTORS REMAIN OBSCURE. HERE, BY SCREENING A SERIES OF METHYLTRANSFERASES AND DEMETHYLASES, WE IDENTIFIED PROTEIN ARGININE METHYLTRANSFERASE 5 (PRMT5) AS AN EFFECTIVE RESTRICTOR OF HBV TRANSCRIPTION AND REPLICATION. IN CELL CULTURE-BASED MODELS FOR HBV INFECTION AND IN LIVER TISSUES OF PATIENTS WITH CHRONIC HBV INFECTION, WE FOUND THAT SYMMETRIC DIMETHYLATION OF ARGININE 3 ON H4 ON CCCDNA WAS A REPRESSIVE MARKER OF CCCDNA TRANSCRIPTION AND WAS REGULATED BY PRMT5 DEPENDING ON ITS METHYLTRANSFERASE DOMAIN. MOREOVER, PRMT5-TRIGGERED SYMMETRIC DIMETHYLATION OF ARGININE 3 ON H4 ON THE CCCDNA MINICHROMOSOME INVOLVED AN INTERACTION WITH THE HBV CORE PROTEIN AND THE BRG1-BASED HUMAN SWI/SNF CHROMATIN REMODELER, WHICH RESULTED IN DOWN-REGULATION OF THE BINDING OF RNA POLYMERASE II TO CCCDNA. IN ADDITION TO THE INHIBITORY EFFECT ON CCCDNA TRANSCRIPTION, PRMT5 INHIBITED HBV CORE PARTICLE DNA PRODUCTION INDEPENDENTLY OF ITS METHYLTRANSFERASE ACTIVITY. FURTHER STUDY REVEALED THAT PRMT5 INTERFERED WITH PREGENOMIC RNA ENCAPSIDATION BY PREVENTING ITS INTERACTION WITH VIRAL POLYMERASE PROTEIN THROUGH BINDING TO THE REVERSE TRANSCRIPTASE-RIBONUCLEASE H REGION OF POLYMERASE, WHICH IS CRUCIAL FOR THE POLYMERASE-PREGENOMIC RNA INTERACTION. CONCLUSION: PRMT5 RESTRICTS HBV REPLICATION THROUGH A TWO-PART MECHANISM INCLUDING EPIGENETIC SUPPRESSION OF CCCDNA TRANSCRIPTION AND INTERFERENCE WITH PREGENOMIC RNA ENCAPSIDATION; THESE FINDINGS IMPROVE THE UNDERSTANDING OF EPIGENETIC REGULATION OF HBV TRANSCRIPTION AND HOST-HBV INTERACTION, THUS PROVIDING NEW INSIGHTS INTO TARGETED THERAPEUTIC INTERVENTION. (HEPATOLOGY 2017;66:398-415).	2017	
                                                                                                                                                                                                                                        
16  3448  33 HYPERMETHYLATION OF THE N-MYC DOWNSTREAM-REGULATED GENE 2 PROMOTER IN PERIPHERAL BLOOD MONONUCLEAR CELLS IS ASSOCIATED WITH LIVER FIBROSIS IN CHRONIC HEPATITIS B. DNA METHYLATION IS A FUNDAMENTAL EPIGENETIC MODIFICATION TO REGULATE GENE EXPRESSION. N-MYC DOWNSTREAM-REGULATED GENE (NDRG) 2 IS A CYTOPLASMIC PROTEIN AND PARTICIPATES IN THE PATHOGENESIS OF LIVER FIBROSIS. IN THIS STUDY, THE MRNA EXPRESSION AND METHYLATION STATUS OF NDRG2 WAS EVALUATED IN PATIENTS WITH CHRONIC HEPATITIS B (CHB). THE STUDY INCLUDED 143 CHB PATIENTS AND 65 NORMAL CONTROLS (NC). THE MRNA EXPRESSION OF NDRG2 IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) WAS DETECTED BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION. THE METHYLATION STATUS OF THE NDRG2 PROMOTER IN PBMCS WAS DETECTED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. THE NDRG2 MRNA LEVEL WAS LOWER IN THE CHB GROUP THAN IN THE NC GROUP (P < 0.001). METHYLATION FREQUENCY OF THE NDRG2 PROMOTER WAS SIGNIFICANTLY HIGHER IN CHB PATIENTS THAN IN THE NC GROUP (52.44% VS. 26.15%, P < 0.001). IMPORTANTLY, THE RELATIVE EXPRESSION LEVELS OF NDRG2 MRNA WERE SIGNIFICANTLY LOWER IN THE METHYLATED GROUP THAN IN THE UNMETHYLATED GROUP IN BOTH CHB PATIENTS AND NC (P < 0.001). FURTHERMORE, A LOWER MRNA LEVEL AND HYPERMETHYLATION OF NDRG2 WERE ASSOCIATED WITH LIVER FIBROSIS AND INFLAMMATION GRADE IN CHB. THE ASPARTATE AMINOTRANSFERASE-TO-PLATELET RATIO INDEX (APRI) SCORE IS WIDELY USED TO PREDICT LIVER FIBROSIS. THE MRNA EXPRESSION LEVELS AND METHYLATION STATUS OF NDRG2 SHOWED A BETTER SCORE COMPARED TO APRI FOR DISCRIMINATING THE SEVERITY OF LIVER FIBROSIS. IN CONCLUSION, HYPERMETHYLATION OF NDRG2 IN PBMCS WAS CORRELATED WITH DECREASED MRNA EXPRESSION AND WITH LIVER FIBROSIS. THE METHYLATION STATUS OF THE NDRG2 PROMOTER IN PBMCS IS A POTENTIAL NONINVASIVE BIOMARKER TO PREDICT THE SEVERITY OF LIVER FIBROSIS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
17  5368  25 RECENT ADVANCES IN THE STUDY OF HEPATITIS B VIRUS COVALENTLY CLOSED CIRCULAR DNA. CHRONIC HEPATITIS B INFECTION IS CAUSED BY HEPATITIS B VIRUS (HBV) AND A TOTAL CURE IS YET TO BE ACHIEVED. THE VIRAL COVALENTLY CLOSED CIRCULAR DNA (CCCDNA) IS THE KEY TO ESTABLISH A PERSISTENT INFECTION WITHIN HEPATOCYTES. CURRENT ANTIVIRAL STRATEGIES HAVE NO EFFECT ON THE PRE-EXISTING CCCDNA RESERVOIR. THEREFORE, THE STUDY OF THE MOLECULAR MECHANISM OF CCCDNA FORMATION IS BECOMING A MAJOR FOCUS OF HBV RESEARCH. THIS REVIEW SUMMARIZES THE CURRENT ADVANCES IN CCCDNA MOLECULAR BIOLOGY AND THE LATEST STUDIES ON THE ELIMINATION OR INACTIVATION OF CCCDNA, INCLUDING THREE MAJOR AREAS: (1) EPIGENETIC REGULATION OF CCCDNA BY HBV X PROTEIN, (2) IMMUNE-MEDIATED DEGRADATION, AND (3) GENOME-EDITING NUCLEASES. ALL THESE ASPECTS PROVIDE CLUES ON HOW TO FINALLY ATTAIN A CURE FOR CHRONIC HEPATITIS B INFECTION.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
18  2842  26 FREQUENT CONCOMITANT EPIGENETIC SILENCING OF SOX1 AND SECRETED FRIZZLED-RELATED PROTEINS (SFRPS) IN HUMAN HEPATOCELLULAR CARCINOMA. BACKGROUND AND AIM: EXCEPT FOR GENETIC MUTATIONS, EPIGENETIC CHANGES ARE ALSO INVOLVED IN THE DEVELOPMENT OF HUMAN CANCERS. RECENTLY, WE HAVE IDENTIFIED SOX1, SRY (SEX DETERMINING REGION Y)-BOX 1, IS HYPERMETHYLATED IN CERVICAL CANCER AND OVARIAN CANCER. THEREFORE, WE INVESTIGATED WHETHER PROMOTER HYPERMETHYLATION OF SOX1 IS COMMON IN HEPATOCELLULAR CARCINOMA (HCC). METHODS: WE USED METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MS-PCR) AND BISULFITE SEQUENCING TO ANALYZE THE METHYALTION LEVEL OF THE SOX1 PROMOTER IN SEVEN HCC CELL LINES, 54 CLINICAL HCCS, 42 CIRRHOTIC LIVERS, 21 LIVERS WITH CHRONIC HEPATITIS, AND 15 CONTROL LIVERS. THEN, WE EMPLOYED QUANTITATIVE MS-PCR (QMSP) TO VALIDATE IN AN INDEPENDENT SET OF SAMPLES (60 PAIRED HCCS AND 30 CONTROL LIVERS). FINALLY, WE USED LUCIFERASE REPORTER AND COLONY FORMATION ASSAY TO CHECK THE EFFECT OF SOX1 IN HCC. RESULTS: PROMOTER METHYLATION OF SOX1 WAS SIGNIFICANTLY FREQUENT IN HCC CELL LINES AND CLINICAL HCCS, CIRRHOTIC LIVERS, BUT NOT IN CONTROL LIVERS (P < 0.0001). THERE IS A SIGNIFICANT CORRELATION BETWEEN DOWNREGULATION OF SOX1 EXPRESSION AND PROMOTER METHYLATION. QMSP RESULTS CONFIRMED THAT PROMOTER HYPERMETHYLATION OF SOX1 IS SIGNIFICANTLY MORE FREQUENT IN HCCS THAN CONTROL LIVERS (P < 0.0001). THE FREQUENCY OF SOX1 METHYLATION IN PATIENTS WITH SECRETED FRIZZLED-RELATED PROTEINS (SFRPS) METHYLATION IS SIGNIFICANTLY HIGHER THAN IN PATIENTS WITHOUT SFRPS METHYLATION (P < 0.0001). FURTHERMORE, ECTOPIC EXPRESSION OF SOX1 COULD SUPPRESS T-CELL FACTOR-DEPENDENT TRANSCRIPTIONAL ACTIVITY AND COLONY FORMATION NUMBER IN HCCS. CONCLUSIONS: CONCOMITANT EPIGENETIC SILENCING OF SOX1 AND SFRPS THROUGH PROMOTER HYPERMETHYLATION IS FREQUENT IN HCCS, AND THIS MIGHT CONTRIBUTE TO ABNORMAL ACTIVATION OF CANONICAL WNT SIGNAL PATHWAY.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
19  6707  33 VIRAL HEPATITIS AND HEPATOCELLULAR CARCINOMA: STATE OF THE ART. VIRAL HEPATITIS IS ONE OF THE MAIN CAUSES LEADING TO HEPATOCELLULAR CARCINOMA (HCC). THE CONTINUED RISE IN INCIDENCE OF HCC SUGGESTS ADDITIONAL FACTORS FOLLOWING INFECTION MAY BE INVOLVED. THIS REVIEW EXAMINES RECENT STUDIES INVESTIGATING THE MOLECULAR MECHANISMS OF CHRONIC HEPATITIS AND ITS ASSOCIATION WITH HEPATOCARCINOGENESIS. HEPATITIS B VIRUS PATIENTS WITH GENOTYPE C DISPLAY AN AGGRESSIVE DISEASE COURSE LEADING TO HCC MORE THAN OTHER GENOTYPES. FURTHERMORE, HEPATITIS B EXCRETORY ANTIGEN (HBEAG) SEEMS TO BE A MORE SENSITIVE PREDICTIVE TUMOR MARKER EXHIBITING A SIX-FOLD HIGHER RELATIVE RISK IN PATIENTS WITH POSITIVE HBSAG AND HBEAG THAN THOSE WITH HBSAG ONLY. SINGLE OR COMBINED MUTATIONS OF VIRAL GENOME CAN PREDICT HCC DEVELOPMENT IN UP TO 80% OF PATIENTS. SEVERAL MUTATIONS IN HBX-GENE ARE RELATED WITH HIGHER HCC INCIDENCE. OVEREXPRESSION OF THE CORE PROTEIN IN HCV LEADS TO HEPATOCELLULAR LIPID ACCUMULATION ASSOCIATED WITH ONCOGENESIS. REDUCED NUMBER AND DECREASED FUNCTIONALITY OF NATURAL KILLER CELLS IN CHRONIC HCV INDIVIDUALS DYSREGULATE THEIR SURVEILLANCE FUNCTION IN TUMOR AND VIRAL CELLS RESULTING IN HCC. FURTHERMORE, HIGH T-CELL IMMUNOGLOBULIN AND MUCIN 3 LEVELS SUPRESS CD8+ T-CELLS, WHICH LEAD TO IMMUNOLOGICAL DYSREGULATION. HEPATITIS D PROMOTES HCC DEVELOPMENT INDIRECTLY VIA MODIFICATIONS TO INNATE IMMUNITY, EPIGENETIC ALTERATIONS AND PRODUCTION OF REACTIVE OXYGEN SPECIES WITH THE LHDAG BEING THE MOST HIGHLY ASSOCIATED WITH HCC DEVELOPMENT. SUMMARIZING THE RESULTS, HBV AND HCV INFECTION REPRESENT THE MOST ASSOCIATED FORMS OF VIRAL HEPATITIS CAUSING HCC. FURTHER STUDIES ARE WARRANTED TO FURTHER IMPROVE THE PREDICTION OF HIGH-RISK PATIENTS AND DEVELOPMENT OF TARGETED THERAPEUTICS PREVENTING THE TRANSITION FROM HEPATIC INFLAMMATION-FIBROSIS TO CANCER.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
20  3190  32 HCV-INDUCED EPIGENETIC CHANGES ASSOCIATED WITH LIVER CANCER RISK PERSIST AFTER SUSTAINED VIROLOGIC RESPONSE. BACKGROUND & AIMS: CHRONIC HEPATITIS C VIRUS (HCV) INFECTION IS AN IMPORTANT RISK FACTOR FOR HEPATOCELLULAR CARCINOMA (HCC). DESPITE EFFECTIVE ANTIVIRAL THERAPIES, THE RISK FOR HCC IS DECREASED BUT NOT ELIMINATED AFTER A SUSTAINED VIROLOGIC RESPONSE (SVR) TO DIRECT-ACTING ANTIVIRAL (DAA) AGENTS, AND THE RISK IS HIGHER IN PATIENTS WITH ADVANCED FIBROSIS. WE INVESTIGATED HCV-INDUCED EPIGENETIC ALTERATIONS THAT MIGHT AFFECT RISK FOR HCC AFTER DAA TREATMENT IN PATIENTS AND MICE WITH HUMANIZED LIVERS. METHODS: WE PERFORMED GENOME-WIDE CHIPMENTATION-BASED CHIP-SEQ AND RNA-SEQ ANALYSES OF LIVER TISSUES FROM 6 PATIENTS WITHOUT HCV INFECTION (CONTROLS), 18 PATIENTS WITH CHRONIC HCV INFECTION, 8 PATIENTS WITH CHRONIC HCV INFECTION CURED BY DAA TREATMENT, 13 PATIENTS WITH CHRONIC HCV INFECTION CURED BY INTERFERON THERAPY, 4 PATIENTS WITH CHRONIC HEPATITIS B VIRUS INFECTION, AND 7 PATIENTS WITH NONALCOHOLIC STEATOHEPATITIS IN EUROPE AND JAPAN. HCV-INDUCED EPIGENETIC MODIFICATIONS WERE MAPPED BY COMPARATIVE ANALYSES WITH MODIFICATIONS ASSOCIATED WITH OTHER LIVER DISEASE ETIOLOGIES. UPA/SCID MICE WERE ENGRAFTED WITH HUMAN HEPATOCYTES TO CREATE MICE WITH HUMANIZED LIVERS AND GIVEN INJECTIONS OF HCV-INFECTED SERUM SAMPLES FROM PATIENTS; MICE WERE GIVEN DAAS TO ERADICATE THE VIRUS. PATHWAYS ASSOCIATED WITH HCC RISK WERE IDENTIFIED BY INTEGRATIVE PATHWAY ANALYSES AND VALIDATED IN ANALYSES OF PAIRED HCC TISSUES FROM 8 PATIENTS WITH AN SVR TO DAA TREATMENT OF HCV INFECTION. RESULTS: WE FOUND CHRONIC HCV INFECTION TO INDUCE SPECIFIC GENOME-WIDE CHANGES IN H3K27AC, WHICH CORRELATED WITH CHANGES IN EXPRESSION OF MRNAS AND PROTEINS. THESE CHANGES PERSISTED AFTER AN SVR TO DAAS OR INTERFERON-BASED THERAPIES. INTEGRATIVE PATHWAY ANALYSES OF LIVER TISSUES FROM PATIENTS AND MICE WITH HUMANIZED LIVERS DEMONSTRATED THAT HCV-INDUCED EPIGENETIC ALTERATIONS WERE ASSOCIATED WITH LIVER CANCER RISK. COMPUTATIONAL ANALYSES ASSOCIATED INCREASED EXPRESSION OF SPHK1 WITH HCC RISK. WE VALIDATED THESE FINDINGS IN AN INDEPENDENT COHORT OF PATIENTS WITH HCV-RELATED CIRRHOSIS (N = 216), A SUBSET OF WHICH (N = 21) ACHIEVED VIRAL CLEARANCE. CONCLUSIONS: IN AN ANALYSIS OF LIVER TISSUES FROM PATIENTS WITH AND WITHOUT AN SVR TO DAA THERAPY, WE IDENTIFIED EPIGENETIC AND GENE EXPRESSION ALTERATIONS ASSOCIATED WITH RISK FOR HCC. THESE ALTERATIONS MIGHT BE TARGETED TO PREVENT LIVER CANCER IN PATIENTS TREATED FOR HCV INFECTION.	2019