1 1730 150 DYSREGULATION OF STEM CELL SIGNALING NETWORK DUE TO GERMLINE MUTATION, SNP, HELICOBACTER PYLORI INFECTION, EPIGENETIC CHANGE AND GENETIC ALTERATION IN GASTRIC CANCER. GENETIC FACTORS, HELICOBACTER PYLORI INFECTION, SALT OVER-UPTAKE, DECREASED VEGETABLE/FRUIT CONSUMPTION, SMOKING, AND METABOLIC SYNDROME ARE RISK FACTORS OF HUMAN GASTRIC CANCER. GERMLINE MUTATIONS OF CDH1 GENE, AND SNPS OF PTPN11 (SHP2), TLR4, IL1B, TNFA, BMP6, GDF15 AND RUNX3 GENES ARE ASSOCIATED WITH GASTRIC CANCER. HELICOBACTER PYLORI ACTIVATES CAGA-SHP2-ERK AND PEPTIDOGLYCAN-NOD1-NFKAPPAB SIGNALING CASCADES IN GASTRIC EPITHELIAL CELLS USING TYPE IV SECRETION SYSTEM, AND ALSO TRAF6-MAP3K7-NFKAPPAB AND TRAF6-MAP3K7-AP-1 SIGNALING CASCADES IN EPITHELIAL AND IMMUNE CELLS THROUGH LIPOPOLYSACCHARIDE RECOGNITION BY TLR2 OR TLR4. IL-1BETA, IL-6, IL-8, TNFALPHA AND IFNGAMMA ARE ELEVATED IN GASTRIC MUCOSA WITH HELICOBACTER PYLORI INFECTION. IL-6 AND TNFALPHA INDUCE UPREGULATION OF WNT5A AND WNT10B, RESPECTIVELY. WNT SIGNALS ARE TRANSDUCED TO BETA-CATENIN-TCF/LEF, RHOA, JNK, PKC, NFAT, AND NLK SIGNALING CASCADES. WNT-BETA-CATENIN-TCF/LEF SIGNALING INDUCES UPREGULATION OF MYC, CCND1, WISP1, FGF20, JAG1 AND DKK1 GENES. NOTCH SIGNALS ARE TRANSDUCED TO CSL-NICD-MAML AND NFKAPPAB SIGNALING CASCADES. FGF SIGNALS ARE TRANSDUCED TO ERK, PI3K-AKT, PKC, AND NFAT SIGNALING CASCADES. HELICOBACTER PYLORI INFECTION INDUCES SHH UPREGULATION IN PARIETAL CELL LINEAGE, WHILE BMP SIGNALS INDUCE IHH UPREGULATION IN PIT CELL LINEAGE. HEDGEHOG SIGNALS INDUCE UPREGULATION OF GLI1, PTCH1, CCND2, FOXL1, JAG2 AND SFRP1 GENES. JAG1 AND JAG2 ACTIVATE NOTCH SIGNALING, WHILE DKK1 AND SFRP1 INHIBIT WNT SIGNALING. STEM CELL SIGNALING NETWORK, CONSISTING OF WNT, NOTCH, FGF, HEDGEHOG AND BMP SIGNALING PATHWAYS, IS ACTIVATED DURING CHRONIC HELICOBACTER PYLORI INFECTION. EPIGENETIC SILENCING OF SFRP1 GENE OCCURS IN THE EARLIER STAGE OF CARCINOGENESIS IN THE STOMACH, WHILE AMPLIFICATION AND OVEREXPRESSION OF FGFR2 GENE IN THE LATER STAGE. DYSREGULATION OF THE STEM CELL SIGNALING NETWORK DUE TO THE ACCUMULATION OF GERMLINE MUTATION, SNP, HELICOBACTER PYLORI INFECTION, EPIGENETIC CHANGE AND GENETIC ALTERATION GIVES RISE TO GASTRIC CANCER. SNP TYPING AND CUSTOM-MADE MICROARRAY ANALYSES ON GENES ENCODING STEM CELL SIGNALING MOLECULES COULD BE UTILIZED FOR THE PERSONALIZED MEDICINE. 2007 2 6758 46 WNT SIGNALING IN STEM CELL BIOLOGY AND REGENERATIVE MEDICINE. WNT FAMILY MEMBERS ARE SECRETED-TYPE GLYCOPROTEINS TO ORCHESTRATE EMBRYOGENESIS, TO MAINTAIN HOMEOSTASIS, AND TO INDUCE PATHOLOGICAL CONDITIONS. FZD1, FZD2, FZD3, FZD4, FZD5, FZD6, FZD7, FZD8, FZD9, FZD10, LRP5, LRP6, AND ROR2 ARE TRANSMEMBRANE RECEPTORS TRANSDUCING WNT SIGNALS BASED ON LIGAND-DEPENDENT PREFERENTIALITY FOR CAVEOLIN- OR CLATHRIN-MEDIATED ENDOCYTOSIS. WNT SIGNALS ARE TRANSDUCED TO CANONICAL PATHWAY FOR CELL FATE DETERMINATION, AND TO NON-CANONICAL PATHWAYS FOR REGULATION OF PLANAR CELL POLARITY, CELL ADHESION, AND MOTILITY. MYC, CCND1, AXIN2, FGF20, WISP1, JAG1, DKK1 AND GLUCAGON ARE TARGET GENES OF CANONICAL WNT SIGNALING CASCADE, WHILE CD44, VIMENTIN AND STX5 ARE TARGET GENES OF NON-CANONICAL WNT SIGNALING CASCADES. HOWEVER, TARGET GENES OF WNT SIGNALING CASCADES ARE DETERMINED IN A CONTEXT-DEPENDENT MANNER DUE TO EXPRESSION PROFILE OF TRANSCRIPTION FACTORS AND EPIGENETIC STATUS. WNT SIGNALING CASCADES NETWORK WITH NOTCH, FGF, BMP AND HEDGEHOG SIGNALING CASCADES TO REGULATE THE BALANCE OF STEM CELLS AND PROGENITOR CELLS. HERE WNT SIGNALING IN EMBRYONIC STEM CELLS, NEURAL STEM CELLS, MESENCHYMAL STEM CELLS, HEMATOPOIETIC STEM CELLS, AND INTESTINAL STEM CELLS WILL BE REVIEWED. WNT3, WNT5A AND WNT10B ARE EXPRESSED IN UNDIFFERENTIATED HUMAN EMBRYONIC STEM CELLS, WHILE WNT6, WNT8B AND WNT10B IN ENDODERM PRECURSOR CELLS. WNT6 IS EXPRESSED IN INTESTINAL CRYPT REGION FOR STEM OR PROGENITOR CELLS. TNF/ALPHA-WNT10B SIGNALING IS A NEGATIVE FEEDBACK LOOP TO MAINTAIN HOMEOSTASIS OF ADIPOSE TISSUE AND GASTROINTESTINAL MUCOSA WITH CHRONIC INFLAMMATION. RECOMBINANT WNT PROTEIN OR WNT MIMETIC (CIRCULAR PEPTIDE, SMALL MOLECULE COMPOUND, OR RNA APTAMER) IN COMBINATION WITH NOTCH MIMETIC, FGF PROTEIN, AND BMP PROTEIN OPENS A NEW WINDOW TO TISSUE ENGINEERING FOR REGENERATIVE MEDICINE. 2008 3 3216 56 HEDGEHOG SIGNALING PATHWAY AND GASTROINTESTINAL STEM CELL SIGNALING NETWORK (REVIEW). HEDGEHOG, BMP/TGFBETA, FGF, WNT AND NOTCH SIGNALING PATHWAYS CONSTITUTE THE STEM CELL SIGNALING NETWORK, WHICH PLAYS A KEY ROLE IN A VARIETY OF PROCESSES, SUCH AS EMBRYOGENESIS, MAINTENANCE OF ADULT TISSUE HOMEOSTASIS, TISSUE REPAIR DURING CHRONIC PERSISTENT INFLAMMATION, AND CARCINOGENESIS. SONIC HEDGEHOG (SHH), INDIAN HEDGEHOG (IHH) AND DESERT HEDGEHOG (DHH) BIND TO PTCH1/PTCH OR PTCH2 RECEPTOR TO RELEASE SMOOTHENED (SMO) SIGNAL TRANSDUCER FROM PATCHED-DEPENDENT SUPPRESSION. SMO THEN ACTIVATES STK36 SERINE/THREONINE KINASE TO STABILIZE GLI FAMILY MEMBERS AND TO PHOSPHORYLATE SUFU FOR NUCLEAR ACCUMULATION OF GLI. HEDGEHOG SIGNALING ACTIVATION LEADS TO GLI-DEPENDENT TRANSCRIPTIONAL ACTIVATION OF TARGET GENES, SUCH AS GLI1, PTCH1, CCND2, FOXL1, JAG2 AND SFRP1. GLI1-DEPENDENT POSITIVE FEEDBACK LOOP COMBINED WITH PTCH1-DEPENDENT NEGATIVE FEEDBACK LOOP GIVES RISE TO TRANSIENT PROLIFERATION OF HEDGEHOG TARGET CELLS. IGUANA HOMOLOGS (DZIP1 AND DZIP1L) AND COSTAL-2 HOMOLOGS (KIF7 AND KIF27) ARE IDENTIFIED BY COMPARATIVE INTEGROMICS. SHH-DEPENDENT PARIETAL CELL PROLIFERATION IS IMPLICATED IN GASTRIC MUCOSAL REPAIR DURING CHRONIC HELICOBACTER PYLORI INFECTION. BMP-RUNX3 SIGNALING INDUCES IHH EXPRESSION IN SURFACE DIFFERENTIATED EPITHELIAL CELLS OF STOMACH AND INTESTINE. HEDGEHOG SIGNALS FROM EPITHELIAL CELLS THEN INDUCES FOXL1-MEDIATED BMP4 UPREGULATION IN MESENCHYMAL CELLS. HEDGEHOG SIGNALING IS FREQUENTLY ACTIVATED IN ESOPHAGEAL CANCER, GASTRIC CANCER AND PANCREATIC CANCER DUE TO TRANSCRIPTIONAL UPREGULATION OF HEDGEHOG LIGANDS AND EPIGENETIC SILENCING OF HHIP1/HHIP GENE, ENCODING THE HEDGEHOG INHIBITOR. HOWEVER, HEDGEHOG SIGNALING IS RARELY ACTIVATED IN COLORECTAL CANCER DUE TO NEGATIVE REGULATION BY THE CANONICAL WNT SIGNALING PATHWAY. HEDGEHOG SIGNALING MOLECULES OR TARGETS, SUCH AS SHH, IHH, HHIP1, PTCH1 AND GLI1, ARE APPLIED AS BIOMARKERS FOR CANCER DIAGNOSTICS, PROGNOSTICS AND THERAPEUTICS. SMALL-MOLECULE INHIBITORS FOR SMO OR STK36 ARE SUITABLE TO BE USED FOR TREATMENT OF HEDGEHOG-DEPENDENT CANCER. 2006 4 6756 38 WNT ANTAGONIST, SFRP1, IS HEDGEHOG SIGNALING TARGET. HEDGEHOG AND WNT SIGNALING PATHWAYS NETWORK TOGETHER DURING EMBRYOGENESIS AND CARCINOGENESIS. HEDGEHOG SIGNALING IN INTESTINAL EPITHELIUM REPRESSES CANONICAL WNT SIGNALING TO RESTRICT EXPRESSION OF WNT TARGET GENES TO STEM OR PROGENITOR CELLS; HOWEVER, THE MECHANISM REMAINS UNCLEAR. THE HEDGEHOG SIGNAL IS TRANSDUCED TO GLI FAMILY TRANSCRIPTION FACTORS THOUGH PATCHED RECEPTOR, SMOOTHENED SIGNAL TRANSDUCER, AND OTHER SIGNALING COMPONENTS, SUCH AS KIF27, KIF7, STK36, SUFU, AND DZIP1. HERE, WE SEARCHED FOR THE GLI-BINDING SITE WITHIN THE PROMOTER REGION OF GENES ENCODING SECRETED-TYPE WNT SIGNAL INHIBITORS, INCLUDING SFRP1, SFRP2, SFRP3, SFRP4, SFRP5, DKK1, DKK2, DKK3, DKK4, AND WIF1. THE GLI-BINDING SITE WAS IDENTIFIED WITHIN THE HUMAN SFRP1 PROMOTER BASED ON BIOINFORMATICS AND HUMAN INTELLIGENCE. THE CHIMPANZEE SFRP1 GENE WAS IDENTIFIED WITHIN THE NW_110515.1 GENOME SEQUENCE. THE GLI-BINDING SITE OF THE HUMAN SFRP1 PROMOTER WAS CONSERVED IN CHIMPANZEE SFRP1, MOUSE SFRP1, AND RAT SFRP1 PROMOTERS. SFRP1 IS THE EVOLUTIONARILY CONSERVED TARGET OF THE HEDGEHOG-GLI SIGNALING PATHWAY. EXPRESSION DOMAIN ANALYSES BASED ON TEXT MINING REVEALED THAT INDIAN HEDGEHOG (IHH), SFRP1, AND WNT6 ARE EXPRESSED IN DIFFERENTIATED INTESTINAL EPITHELIAL CELLS, MESENCHYMAL CELLS, AND STEM/PROGENITOR CELLS, RESPECTIVELY. HEDGEHOG IS SECRETED FROM DIFFERENTIATED EPITHELIAL CELLS TO INDUCE SFRP1 EXPRESSION IN MESENCHYMAL CELLS, WHICH KEEPS DIFFERENTIATED EPITHELIAL CELLS AWAY FROM THE EFFECTS OF CANONICAL WNT SIGNALING. THESE FACTS INDICATE THAT SFRP1 IS THE HEDGEHOG TARGET TO CONFINE CANONICAL WNT SIGNALING WITHIN STEM OR PROGENITOR CELLS. THEREFORE, EPIGENETIC CPG HYPERMETHYLATION OF THE SFRP1 PROMOTER DURING CHRONIC PERSISTENT INFLAMMATION AND AGING LEADS TO THE OCCURRENCE OF GASTROINTESTINAL CANCERS, SUCH AS COLORECTAL CANCER AND GASTRIC CANCER, THROUGH THE BREAKDOWN OF HEDGEHOG-DEPENDENT WNT SIGNAL INHIBITION. 2006 5 2811 45 FGFR2-RELATED PATHOGENESIS AND FGFR2-TARGETED THERAPEUTICS (REVIEW). FGFR2 GENE AT HUMAN CHROMOSOME 10Q26 ENCODES FGFR2B AND FGFR2C ISOFORMS FUNCTIONING AS FGF RECEPTORS WITH DISTINCT EXPRESSION DOMAIN AND LIGAND SPECIFICITY. FGFR2 PLAYS ONCOGENIC AND ANTI-ONCOGENIC ROLES IN A CONTEXT-DEPENDENT MANNER. SINGLE NUCLEOTIDE POLYMORPHISMS (SNPS) WITHIN INTRON 2 OF FGFR2 GENE ARE ASSOCIATED WITH BREAST CANCER THROUGH ALLELIC FGFR2 UPREGULATION. MISSENSE MUTATIONS OR COPY NUMBER GAINS OF FGFR2 GENE OCCUR IN BREAST CANCER AND GASTRIC CANCER TO ACTIVATE FGFR2 SIGNALING. ABERRANT FGFR2 SIGNALING ACTIVATION INDUCES PROLIFERATION AND SURVIVAL OF TUMOR CELLS. THE CLASS SWITCH FROM FGFR2B TO FGFR2C OCCURS DURING PROGRESSION OF PROSTATE CANCER AND BLADDER CANCER BECAUSE OF SPLICEOSOME DYSREGULATION. IN ADDITION, EPIDERMAL FGFR2B KNOCKOUT MICE SHOW INCREASED SENSITIVITY TO CHEMICAL CARCINOGENESIS PARTLY DUE TO THE FAILURE OF NFE2L2 (NRF2)-MEDIATED DETOXIFICATION OF REACTIVE OXYGEN SPECIES (ROS). LOSS OF FGFR2B SIGNALING INDUCES EPITHELIAL-TO-MESENCHYMAL TRANSITION (EMT) AND UNRULY ROS. FGFR2 SIGNALING DYSREGULATION DUE TO THE ACCUMULATION OF EPIGENETIC MODIFICATIONS AND GENETIC ALTERATIONS DURING CHRONIC INFLAMMATION, SMOKING, INCREASED CALORIC UPTAKE, AND DECREASED EXERCISE LEADS TO CARCINOGENESIS. PD173074, SU5402, AZD2171, AND KI23057 ARE SMALL-MOLECULE FGFR INHIBITORS. HUMAN ANTIBODY, PEPTIDE MIMETIC, RNA APTAMER, SIRNA, AND SYNTHETIC MICRORNA (MIRNA) ARE EMERGING TECHNOLOGIES TO BE APPLIED FOR CANCER THERAPEUTICS TARGETED TO FGFR2. BECAUSE NOVEL SEQUENCE TECHNOLOGY AND PETA-SCALE SUPER-COMPUTER ARE OPENING UP THE SEQUENCE ERA FOLLOWING THE GENOME ERA, PERSONALIZED MEDICINE PRESCRIBING TARGETED DRUGS BASED ON GERMLINE AND/OR SOMATIC GENOMIC INFORMATION IS COMING REALITY. APPLICATION OF FGFR2 INHIBITORS FOR CANCER TREATMENT IN PATIENTS WITH FGFR2 MUTATION OR GENE AMPLIFICATION IS BENEFICIAL; HOWEVER, THAT FOR CANCER PREVENTION IN PEOPLE WITH FGFR2 RISK ALLELE MIGHT BE DISADVANTAGEOUS DUE TO THE IMPEDIMENT OF A CYTOPROTECTIVE MECHANISM AGAINST OXIDATIVE STRESS. 2009 6 4162 26 MECP2 REGULATES PTCH1 EXPRESSION THROUGH DNA METHYLATION IN RHEUMATOID ARTHRITIS. RHEUMATOID ARTHRITIS (RA) IS A CHRONIC AUTOIMMUNE INFLAMMATORY DISEASE, IN WHICH PATHOGENESIS IS NOT CLEAR. MANY RESEARCH DEMONSTRATED THAT FIBROBLAST-LIKE SYNOVIOCYTES (FLSS) PLAY A KEY ROLE IN RA PATHOGENESIS, JOIN IN THE CARTILAGE INJURY AND HYPERPLASIA OF THE SYNOVIUM, AND CONTRIBUTE TO THE RELEASE OF INFLAMMATORY CYTOKINES. WE USED ADJUVANT ARTHRITIS (AA) RATS AS RA ANIMAL MODELS. THE METHYL-CPG-BINDING PROTEIN 2 (MECP2) ENABLES THE SUPPRESSED CHROMATIN STRUCTURE TO BE SELECTIVELY DETECTED IN AA FLSS. OVEREXPRESSION OF THIS PROTEIN LEADS TO AN INCREASE OF INTEGRAL METHYLATION LEVELS. SOME RESEARCH HAS CONFIRMED THE HEDGEHOG (HH) SIGNALING PATHWAY PLAYS AN IMPORTANT ROLE IN RA PATHOGENESIS; FURTHERMORE, PATCHED 1 (PTCH1) IS A NEGATIVE FRACTION OF HH SIGNALING PATHWAY. WE USED 5-AZA-2'-DEOXYCYTIDINE (5-AZADC) AS DNA METHYLATION INHIBITOR. IN OUR RESEARCH, WE FOUND MECP2 REDUCED PTCH1 EXPRESSION IN AA FLSS; 5-AZADC OBSTRUCTED THE LOSS OF PTCH1 EXPRESSION. 5-AZADC, TREATMENT OF AA FLSS, ALSO BLOCKS THE RELEASE OF INFLAMMATORY CYTOKINES. IN ORDER TO PROBE THE POTENTIAL MOLECULAR MECHANISM, WE ASSUMED THE EPIGENETIC PARTICIPATION IN THE REGULATION OF PTCH1. RESULTS DEMONSTRATED THAT PTCH1 HYPERMETHYLATION IS RELATED TO THE PERSISTENT FLS ACTIVATION AND INFLAMMATION IN AA RATS. KNOCKDOWN OF MECP2 USING SMALL-INTERFERING RNA TECHNIQUE ADDED PTCH1 EXPRESSION IN AA FLSS. OUR RESULTS INDICATE THAT DNA METHYLATION MAY OFFER MOLECULE MECHANISMS, AND THE REDUCED PTCH1 METHYLATION LEVEL COULD REGULATE INFLAMMATION THROUGH KNOCKDOWN OF MECP2. GRAPHICAL ABSTRACT PTCH1 IS AN INHIBITORY PROTEIN OF THE HEDGEHOG SIGNALING PATHWAY. INCREASED EXPRESSION OF PTCH1 CAN INHIBIT THE EXPRESSION OF GLI1 AND SHH, THEREBY INHIBITING THE ACTIVATION OF HEDGEHOG SIGNALING PATHWAY. INACTIVATED HEDGEHOG SIGNALING PATHWAY INHIBITS THE SECRETION OF IL-6 AND TNF-ALPHA. MECP2 MEDIATES HYPERMETHYLATION OF PTCH1 GENE AND DECREASES THE EXPRESSION OF PTCH1 PROTEIN, THUS ACTIVATING HEDGEHOG SIGNALING PATHWAY AND INCREASING SECRETION OF IL-6 AND TNF-ALPHA. 2017 7 1393 22 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 8 6441 18 THERAPEUTIC APPROACHES FOR NONALCOHOLIC FATTY LIVER DISEASE: ESTABLISHED TARGETS AND DRUGS. NONALCOHOLIC FATTY LIVER DISEASE (NAFLD), AS A MULTISYSTEMIC DISEASE, IS THE MOST PREVALENT CHRONIC LIVER DISEASE CHARACTERIZED BY EXTREMELY COMPLEX PATHOGENIC MECHANISMS AND MULTIFACTORIAL ETIOLOGY, WHICH OFTEN DEVELOPS AS A CONSEQUENCE OF OBESITY, METABOLIC SYNDROME. PATHOPHYSIOLOGICAL MECHANISMS INVOLVED IN THE DEVELOPMENT OF NAFLD INCLUDE DIET, OBESITY, INSULIN RESISTANCE (IR), GENETIC AND EPIGENETIC DETERMINANTS, INTESTINAL DYSBIOSIS, OXIDATIVE/NITROSATIVE STRESS, AUTOPHAGY DYSREGULATION, HEPATIC INFLAMMATION, GUT-LIVER AXIS, GUT MICROBES, IMPAIRED MITOCHONDRIAL METABOLISM AND REGULATION OF HEPATIC LIPID METABOLISM. SOME OF THE NEW DRUGS FOR THE TREATMENT OF NAFLD ARE INTRODUCED HERE. ALL OF THEM ACHIEVE THERAPEUTIC OBJECTIVES BY INTERFERING WITH CERTAIN PATHOPHYSIOLOGICAL PATHWAYS OF NAFLD, INCLUDING FIBROBLAST GROWTH FACTORS (FGF) ANALOGUES, PEROXISOME PROLIFERATOR-ACTIVATED RECEPTORS (PPARS) AGONISTS, GLUCAGON-LIKE PEPTIDE-1 (GLP-1) AGONISTS, G PROTEIN-COUPLED RECEPTORS (GPCRS), SODIUM-GLUCOSE COTRANSPORTER-2 INHIBITORS (SGLT-2I), FARNESOID X RECEPTOR (FXR), FATTY ACID SYNTHASE INHIBITOR (FASNI), ANTIOXIDANTS, ETC. THIS REVIEW DESCRIBES SOME PATHOPHYSIOLOGICAL MECHANISMS OF NAFLD AND ESTABLISHED TARGETS AND DRUGS. 2023 9 1483 23 DKK1 IS EPIGENETICALLY DOWNREGULATED BY PROMOTER METHYLATION AND INHIBITS BILE ACID-INDUCED GASTRIC INTESTINAL METAPLASIA. DICKKOPF-RELATED PROTEIN 1 (DKK1) IS ESSENTIAL TO GASTRIC CANCER AS AN INHIBITOR OF WNT SIGNALING. GASTRIC INTESTINAL METAPLASIA (GIM) IS AN IMPORTANT PRECANCEROUS LESION OF GASTRIC CANCER THAT CAN BE ACTIVATED BY BILE ACID REFLUX AND CHRONIC INFLAMMATION. HOWEVER, THE EXACT MECHANISM OF DKK1 IN BILE ACID-INDUCED GIM HAS NOT BEEN COMPLETELY ELUCIDATED. WE AIMED TO EXPLORE THE EPIGENETIC ALTERATIONS AND BIOLOGICAL FUNCTIONS OF DKK1 IN THE DEVELOPMENT OF GIM. IN THE PRESENT STUDY, BILE ACID WAS FOUND TO INDUCE THE EXPRESSION OF INTESTINAL MARKERS IN GASTRIC EPITHELIAL CELLS, WHEREAS DKK1 WAS DOWNREGULATED IN RESPONSE TO BILE ACID STIMULATION. THE MRNA AND PROTEIN EXPRESSION LEVELS OF DKK1 WERE DECREASED IN GIM TISSUES AS EVIDENCED BY QRT-PCR AND IMMUNOHISTOCHEMICAL STAINING. SURPRISINGLY, THE METHYLATION OF THE DKK1 PROMOTER INCREASED IN GIM TISSUES, AND WE DISCOVERED 28 DIFFERENTIAL METHYLATION SITES OF THE DKK1 PROMOTER IN GIM TISSUES. BILE ACID WAS ABLE TO INDUCE THE PARTIAL METHYLATION OF THE DKK1 PROMOTER, WHILE 5-AZA COULD INCREASE DKK1 EXPRESSION AS WELL AS DECREASE INTESTINAL MARKERS EXPRESSION IN GASTRIC EPITHELIAL CELLS. IN CONCLUSION, THE PROMOTER METHYLATION AND DOWNREGULATION OF DKK1 MIGHT PLAY IMPORTANT ROLES IN THE DEVELOPMENT OF GIM, ESPECIALLY BILE ACID-INDUCED GIM. 2020 10 2809 22 FGF1 AMELIORATES OBESITY-ASSOCIATED HEPATIC STEATOSIS BY REVERSING IGFBP2 HYPERMETHYLATION. OBESITY IS A MAJOR CONTRIBUTING FACTOR FOR METABOLIC-ASSOCIATED FATTY LIVER DISEASE (MAFLD). FIBROBLAST GROWTH FACTOR (FGF) 1 IS THE FIRST PARACRINE FGF FAMILY MEMBER IDENTIFIED TO EXHIBIT PROMISING METABOLIC REGULATORY PROPERTIES CAPABLE OF CONFERRING GLUCOSE-LOWERING AND INSULIN-SENSITIZING EFFECT. THIS STUDY EXPLORES THE ROLE AND MOLECULAR UNDERPINNINGS OF FGF1 IN OBESITY-ASSOCIATED HEPATIC STEATOSIS. IN A MOUSE HIGH-FAT DIET (HFD)-INDUCED MAFLD MODEL, CHRONIC TREATMENT WITH RECOMBINANT FGF1(RFGF1) WAS FOUND TO EFFECTIVELY REDUCE THE SEVERITY OF INSULIN RESISTANCE, HYPERLIPIDEMIA, AND INFLAMMATION. FGF1 TREATMENT DECREASED LIPID ACCUMULATION IN THE MOUSE LIVER AND PALMITIC ACID-TREATED AML12 CELLS. THESE EFFECTS WERE ASSOCIATED WITH DECREASED MATURE FORM SREBF1 EXPRESSION AND ITS TARGET GENES FASN AND SCD1. INTERESTINGLY, WE UNCOVERED THAT RFGF1 SIGNIFICANTLY INDUCED IGFBP2 EXPRESSION AT BOTH MRNA AND PROTEIN LEVELS IN HFD-FED MOUSE LIVERS AND CULTURED HEPATOCYTES TREATED WITH PALMITIC ACID. ADENO-ASSOCIATED VIRUS-MEDIATED IGFBP2 SUPPRESSION SIGNIFICANTLY DIMINISHED THE THERAPEUTIC BENEFIT OF RFGF1 ON MAFLD-ASSOCIATED PHENOTYPES, INDICATING THAT IGFBP2 PLAYS A CRUCIAL ROLE IN THE FGF1-MEDIATED REDUCTION OF HEPATIC STEATOSIS. FURTHER ANALYSIS REVEALED THAT RFGF1 TREATMENT REDUCES THE RECRUITMENT OF DNA METHYLTRANSFERASE 3 ALPHA TO THE IGFBP2 GENOMIC LOCUS, LEADING TO DECREASED IGFBP2 GENE METHYLATION AND INCREASED MRNA AND PROTEIN EXPRESSION. COLLECTIVELY, OUR FINDINGS REVEAL FGF1 MODULATION OF LIPID METABOLISM VIA EPIGENETIC REGULATION OF IGFBP2 EXPRESSION, AND UNRAVEL THE THERAPEUTIC POTENTIAL OF THE FGF1-IGFBP2 AXIS IN METABOLIC DISEASES ASSOCIATED WITH OBESITY. 2023 11 3396 31 HOST SHP1 PHOSPHATASE ANTAGONIZES HELICOBACTER PYLORI CAGA AND CAN BE DOWNREGULATED BY EPSTEIN-BARR VIRUS. MOST IF NOT ALL GASTRIC CANCERS ARE ASSOCIATED WITH CHRONIC INFECTION OF THE STOMACH MUCOSA WITH HELICOBACTER PYLORI CAGA-POSITIVE STRAINS(1-4). APPROXIMATELY 10% OF GASTRIC CANCERS ALSO HARBOUR EPSTEIN-BARR VIRUS (EBV) IN THE CANCER CELLS(5,6). FOLLOWING DELIVERY INTO GASTRIC EPITHELIAL CELLS VIA TYPE IV SECRETION(7,8), THE CAGA-ENCODED CAGA PROTEIN UNDERGOES TYROSINE PHOSPHORYLATION ON THE GLU-PRO-ILE-TYR-ALA (EPIYA) MOTIFS INITIALLY BY SRC FAMILY KINASES (SFKS) AND THEN BY C-ABL(9,10). TYROSINE-PHOSPHORYLATED CAGA BINDS TO THE PRO-ONCOGENIC PROTEIN TYROSINE PHOSPHATASE SHP2 AND THEREBY DEREGULATES THE PHOSPHATASE ACTIVITY(11,12), WHICH HAS BEEN CONSIDERED TO PLAY AN IMPORTANT ROLE IN GASTRIC CARCINOGENESIS(13). HERE WE SHOW THAT THE SHP2 HOMOLOGUE SHP1 INTERACTS WITH CAGA INDEPENDENTLY OF THE EPIYA MOTIF. THE INTERACTION POTENTIATES THE PHOSPHATASE ACTIVITY OF SHP1 THAT DAMPENS THE ONCOGENIC ACTION OF CAGA BY DEPHOSPHORYLATING THE CAGA EPIYA MOTIFS. IN VITRO INFECTION OF GASTRIC EPITHELIAL CELLS WITH EBV INDUCES SHP1 PROMOTER HYPERMETHYLATION, WHICH STRENGTHENS PHOSPHORYLATION-DEPENDENT CAGA ACTION VIA EPIGENETIC DOWNREGULATION OF SHP1 EXPRESSION. CLINICAL SPECIMENS OF EBV-POSITIVE GASTRIC CANCERS ALSO EXHIBIT SHP1 HYPERMETHYLATION WITH REDUCED SHP1 EXPRESSION. THE RESULTS REVEAL THAT SHP1 IS THE LONG-SOUGHT PHOSPHATASE THAT CAN ANTAGONIZE CAGA. AUGMENTED H. PYLORI CAGA ACTIVITY, VIA SHP1 INHIBITION, MIGHT ALSO CONTRIBUTE TO THE DEVELOPMENT OF EBV-POSITIVE GASTRIC CANCER. 2016 12 3642 28 INCREASED H3K4ME3 METHYLATION AND DECREASED MIR-7113-5P EXPRESSION LEAD TO ENHANCED WNT/BETA-CATENIN SIGNALING IN IMMUNE CELLS FROM PTSD PATIENTS LEADING TO INFLAMMATORY PHENOTYPE. BACKGROUND: POSTTRAUMATIC STRESS DISORDER (PTSD) IS A PSYCHIATRIC DISORDER ACCOMPANIED BY CHRONIC PERIPHERAL INFLAMMATION. WHAT TRIGGERS INFLAMMATION IN PTSD IS CURRENTLY UNCLEAR. IN THE PRESENT STUDY, WE IDENTIFIED POTENTIAL DEFECTS IN SIGNALING PATHWAYS IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) FROM INDIVIDUALS WITH PTSD. METHODS: RNASEQ (5 SAMPLES EACH FOR CONTROLS AND PTSD), CHIPSEQ (5 SAMPLES EACH) AND MIRNA ARRAY (6 SAMPLES EACH) WERE USED IN COMBINATION WITH BIOINFORMATICS TOOLS TO IDENTIFY DYSREGULATED GENES IN PBMCS. REAL TIME QRT-PCR (24 SAMPLES EACH) AND IN VITRO ASSAYS WERE EMPLOYED TO VALIDATE OUR PRIMARY FINDINGS AND HYPOTHESIS. RESULTS: BY RNA-SEQ ANALYSIS OF PBMCS, WE FOUND THAT WNT SIGNALING PATHWAY WAS UPREGULATED IN PTSD WHEN COMPARED TO NORMAL CONTROLS. SPECIFICALLY, WE FOUND INCREASED EXPRESSION OF WNT10B IN THE PTSD GROUP WHEN COMPARED TO CONTROLS. OUR FINDINGS WERE CONFIRMED USING NCBI'S GEO DATABASE INVOLVING A LARGER SAMPLE SIZE. ADDITIONALLY, IN VITRO ACTIVATION STUDIES REVEALED THAT ACTIVATED BUT NOT NAIVE PBMCS FROM CONTROL INDIVIDUALS EXPRESSED MORE IFNGAMMA IN THE PRESENCE OF RECOMBINANT WNT10B SUGGESTING THAT WNT SIGNALING PLAYED A CRUCIAL ROLE IN EXACERBATING INFLAMMATION. NEXT, WE INVESTIGATED THE MECHANISM OF INDUCTION OF WNT10B AND FOUND THAT INCREASED EXPRESSION OF WNT10B MAY RESULT FROM EPIGENETIC MODULATION INVOLVING DOWNREGULATION OF HSA-MIR-7113-5P WHICH TARGETED WNT10B. FURTHERMORE, WE ALSO OBSERVED THAT WNT10B OVEREXPRESSION WAS LINKED TO HIGHER EXPRESSION OF H3K4ME3 HISTONE MODIFICATION AROUND THE PROMOTOR OF WNT10B. ADDITIONALLY, KNOCKDOWN OF HISTONE DEMETHYLASE SPECIFIC TO H3K4ME3, USING SIRNA, LED TO INCREASED EXPRESSION OF WNT10B PROVIDING CONCLUSIVE EVIDENCE THAT H3K4ME3 INDEED CONTROLLED WNT10B EXPRESSION. CONCLUSIONS: IN SUMMARY, OUR DATA DEMONSTRATE FOR THE FIRST TIME THAT WNT SIGNALING PATHWAY IS UPREGULATED IN PBMCS OF PTSD PATIENTS RESULTING FROM EPIGENETIC CHANGES INVOLVING MICRORNA DYSREGULATION AND HISTONE MODIFICATIONS, WHICH IN TURN MAY PROMOTE THE INFLAMMATORY PHENOTYPE IN SUCH CELLS. 2020 13 2437 21 EPIGENETIC SILENCING OF SONIC HEDGEHOG ELICITS ANTITUMOR IMMUNE RESPONSE AND SUPPRESSES TUMOR GROWTH BY INHIBITING THE HEDGEHOG SIGNALING PATHWAY IN METASTATIC SPINE TUMORS IN SPRAGUE-DAWLEY RATS. BACKGROUND: PATIENTS WITH METASTATIC SPINE TUMORS MAY SUFFER FROM PAIN OR NEUROLOGIC DEFICIT, AND THE DISEASE MAY BE DETECTED IN PATIENTS WITH A KNOWN MALIGNANCY. SONIC HEDGEHOG (SHH) HAS RECEIVED SPECIAL ATTENTION DUE TO ITS ROLE IN CANCERS. THEREFORE, THIS STUDY INVESTIGATED THE EFFECTS OF EPIGENETIC SILENCING OF SHH ON ANTITUMOR IMMUNE RESPONSE AND TUMOR GROWTH BY REGULATING THE HEDGEHOG (HH) SIGNALING PATHWAY IN METASTATIC SPINE TUMORS. METHODS: RAT MODELS OF METASTATIC SPINE TUMORS WERE SUCCESSFULLY ESTABLISHED. WE FIRST CALCULATED THE TUMOR VOLUME AND THE INHIBITION RATE OF TUMOR GROWTH TO INVESTIGATE THE EFFECT OF SHH ON TUMOR GROWTH. AFTERWARDS, IMMUNOHISTOCHEMISTRY WAS USED TO DETERMINE WHETHER PROLIFERATION WAS DELAYED BY SHH DEPLETION, AND THE 3-(4,5-DIMETHYLTHIAZOL-2-YL)-2,5-DIPHENYLTETRAZOLIUM BROMIDE ASSAY WAS CONDUCTED TO TEST THE CHANGES IN THE LYMPHOCYTE TRANSFORMATION RATE IN THE SPLEEN TRIGGERED BY SHH SILENCING. THEN, THE INFLUENCE OF SHH DEPLETION ON IMMUNE FUNCTION WAS INVESTIGATED. LATER, QUANTITATIVE REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION AND WESTERN BLOT ASSAY WERE PERFORMED TO EXPLORE THE HH SIGNALING PATHWAY-RELATED FACTORS. FINALLY, WE ADDED THE HH SIGNALING PATHWAY INHIBITOR, GDC-0449, TO CONFIRM THE ROLE OF THE PATHWAY IN TUMOR PROGRESSION. RESULTS: INITIALLY, WE OBSERVED THAT SHH DEPLETION WAS A NEGATIVE FACTOR FOR TUMOR GROWTH. AFTERWARDS, IT WAS REVEALED THAT EPIGENETIC SILENCING OF SHH SERVED AS AN INHIBITOR FACTOR FOR THE FUNCTION OF SPLEEN LYMPHOCYTE TRANSFORMATION AND INFLAMMATION WHILE PROMOTING ANTITUMOR IMMUNE FUNCTION. CONCLUSION: OUR PRELIMINARY RESULTS INDICATE THAT EPIGENETIC SILENCING OF SHH ELICITS AN ANTITUMOR IMMUNE RESPONSE AND SUPPRESSES TUMOR GROWTH BY INHIBITING THE HH SIGNALING PATHWAY IN METASTATIC SPINE TUMORS. 2018 14 6662 21 UPREGULATION OF FZD5 IN EOSINOPHILIC CHRONIC RHINOSINUSITIS WITH NASAL POLYPS BY EPIGENETIC MODIFICATION. EOSINOPHILIC CHRONIC RHINOSINUSITIS WITH NASAL POLYPS (CRSWNP) IS ONE OF THE MOST CHALLENGING PROBLEMS IN CLINICAL RHINOLOGY. FZD5 IS A RECEPTOR FOR WNT5A, AND ITS COMPLEX WITH WNT5A CONTRIBUTES TO ACTIVATING INFLAMMATION AND TISSUE MODIFICATION. NASAL POLYPS AND EOSINOPHIL/NON-EOSINOPHIL COUNTS ARE REPORTED TO BE DIRECTLY CORRELATED. THIS STUDY INVESTIGATED THE EXPRESSION AND DISTRIBUTION OF FZD5, AND THE ROLE OF EOSINOPHIL INFILTRATION AND FZD5 IN EOSINOPHILIC CRSWNP PATHOGENESIS. THE PROGNOSTIC ROLE OF EOSINOPHIL LEVELS WAS EVALUATED IN SEVEN PATIENTS WITH CRSWNP. FIFTEEN PATIENTS WITH CRS WERE CLASSIFIED BASED ON THE PERCENTAGE OF EOSINOPHILS IN NASAL POLYP TISSUE. METHYLATED GENES WERE DETECTED USING METHYL-CPG-BINDING DOMAIN SEQUENCING, AND QRT-PCR AND IMMUNOHISTOCHEMISTRY WERE USED TO DETECT FZD5 EXPRESSION IN NASAL POLYP TISSUE SAMPLES. THE RESULTS SHOWED THAT MRNA EXPRESSION OF FZD5 WAS UPREGULATED IN NASAL POLYPS. FZD5 EXPRESSION WAS SIGNIFICANTLY HIGHER IN NASAL POLYP SAMPLES FROM PATIENTS WITH EOSINOPHILIC CRSWNP THAN IN THOSE FROM PATIENTS WITH NON-EOSINOPHILIC CRSWNP, AS INDICATED BY IMMUNOHISTOCHEMISTRY. FURTHERMORE, INFLAMMATORY CYTOKINE LEVELS WERE HIGHER IN EOSINOPHILIC CRSWNP-DERIVED EPITHELIAL CELLS THAN IN NORMAL TISSUES. IN CONCLUSION, FZD5 EXPRESSION IN NASAL MUCOSAL EPITHELIAL CELLS IS CORRELATED WITH INFLAMMATORY CELLS AND MIGHT PLAY A ROLE IN THE PATHOGENESIS OF EOSINOPHILIC CRSWNP. 2019 15 2379 30 EPIGENETIC REGULATION OF WNT PATHWAY ANTAGONISTS IN HUMAN GLIOBLASTOMA MULTIFORME. EPIGENETIC INACTIVATION OF TUMOR SUPPRESSOR GENES IS COMMON IN HUMAN CANCER. USING A LARGE-SCALE WHOLE-GENOME APPROACH IN AN EARLIER STUDY, THE AUTHORS IDENTIFIED EPIGENETICALLY SILENCED GENES WITH POTENTIAL TUMOR SUPPRESSOR FUNCTION IN GLIOBLASTOMA (GBM). THREE GENES IDENTIFIED IN THIS ANALYSIS-DKK1, SFRP1, AND WIF1-ARE POTENT INHIBITORS OF THE WNT SIGNAL TRANSDUCTION PATHWAY. HERE, THE AUTHORS CONFIRM DECREASED EXPRESSION OF THESE GENES IN GBM TUMOR TISSUE SAMPLES RELATIVE TO NONTUMOR BRAIN TISSUE SAMPLES USING REAL-TIME PCR. THEY THEN SHOW THAT EXPRESSION OF ALL 3 GENES IS RESTORED IN T98 GBM CELLS BY TREATMENT WITH THE HISTONE DEACETYLASE INHIBITOR TRICHOSTATIN A (TSA), BUT ONLY DKK1 EXPRESSION IS RESTORED BY TREATMENT WITH THE DEMETHYLATING AGENT 5-AZACYTIDINE. BISULFITE SEQUENCING DID NOT REVEAL SIGNIFICANT METHYLATION IN THE PROMOTER REGION OF DKK1, WHEREAS HISTONE ACETYLATION AND CHROMATIN ACCESSIBILITY INCREASED SIGNIFICANTLY FOR ALL 3 GENES AFTER TSA TREATMENT. ECTOPIC EXPRESSION OF DKK1 SIGNIFICANTLY REDUCES COLONY FORMATION AND INCREASES CHEMOTHERAPY-INDUCED APOPTOSIS IN T98 CELLS. ECTOPIC EXPRESSION OF THE CANONICAL WNT PATHWAY INHIBITORS WIF1 AND SFRP1 SHOWS A RELATIVE LACK OF RESPONSE. CHRONIC WNT3A STIMULATION ONLY PARTIALLY REVERSES GROWTH SUPPRESSION AFTER DKK1 REEXPRESSION, WHEREAS A SPECIFIC INHIBITOR OF THE JNK PATHWAY SIGNIFICANTLY REVERSES THE EFFECT OF DKK1 REEXPRESSION ON COLONY FORMATION AND APOPTOSIS IN T98 CELLS. THESE RESULTS SUPPORT A POTENTIAL GROWTH-SUPPRESSIVE FUNCTION FOR EPIGENETICALLY SILENCED DKK1 IN GBM AND SUGGEST THAT DKK1 RESTORATION COULD MODULATE WNT SIGNALING THROUGH BOTH CANONICAL AND NONCANONICAL PATHWAYS. 2010 16 6529 26 TRANSCRIPTIONAL DOWN-REGULATION OF THE WNT ANTAGONIST SFRP1 IN HAEMATOPOIETIC CELLS OF PATIENTS WITH DIFFERENT RISK TYPES OF MDS. SECRETED FRIZZLED RELATED PROTEIN 1 (SFRP1) IS AN EXTRACELLULAR ANTAGONIST OF THE WNT SIGNALLING PATHWAY THAT PLAYS AN IMPORTANT ROLE IN THE PATHOGENESIS OF SOLID TUMOURS AND HAEMATOPOIETIC MALIGNANCIES. SFRP1 HAS BEEN OBSERVED TO BE TRANSCRIPTIONALLY DOWN-REGULATED DUE TO HYPERMETHYLATION IN ACUTE AND CHRONIC LEUKAEMIA, BUT SO FAR NOT IN MYELODYSPLASTIC SYNDROME (MDS). MOREOVER, IT HAS BEEN SHOWN THAT THE EPIGENETIC INACTIVATION OF SFRP1 CORRELATES WITH AN OVEREXPRESSION OF THE WNT RECEPTOR FRIZZLED 3 (FZD3) IN ACUTE LEUKAEMIA. USING REAL-TIME QUANTITATIVE REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION (RT-PCR) WE EXAMINED MRNA EXPRESSION OF SFRP1 AND FZD3 IN BONE MARROW CELLS DERIVED FROM 121 PATIENTS WITH DIFFERENT RISK TYPES OF MDS, ACUTE MYELOID LEUKAEMIA (AML) AND ACUTE LYMPHOBLASTIC LEUKAEMIA (ALL). WE EMPLOYED PYROSEQUENCING TO QUANTIFY PROMOTER DNA METHYLATION IN MDS AND ACUTE LEUKAEMIA. WE DETECTED SIGNIFICANT LOWER MRNA TRANSCRIPTION OF SFRP1 IN MDS COMPARED TO HEALTHY INDIVIDUALS. HOWEVER, DNA SEQUENCE MUTATIONS OR FREQUENT ELEVATED DNA METHYLATION LEVELS OF THE SFRP1 PROMOTER COULD NOT BE OBSERVED IN MDS BUT IN AML AND ALL AS PREVIOUSLY REPORTED. THE EXPRESSION LEVELS OF FZD3 WERE UP-REGULATED IN BOTH ACUTE LEUKAEMIA AND MDS. OUR DATA SHOW A SIGNIFICANT TRANSCRIPTIONAL DOWN-REGULATION OF SFRP1 AS A COMMON EVENT IN AML, ALL AND - AS DEMONSTRATED FOR THE FIRST TIME - IN MDS. AN INACTIVATION OF SFRP1 AND THE TRANSCRIPTIONAL UP-REGULATION OF FZD3 SEEM TO BE ASSOCIATED WITH AN ACTIVATION OF THE WNT SIGNALLING PATHWAY IN THESE HAEMATOPOIETIC DISEASES. 2010 17 1632 20 DNMTS ARE INVOLVED IN TGF-BETA1-INDUCED EPITHELIAL-MESENCHYMAL TRANSITIONS IN AIRWAY EPITHELIAL CELLS. CHRONIC RHINOSINUSITIS (CRS) PATHOGENESIS IS CLOSELY RELATED TO TISSUE REMODELING, INCLUDING EPITHELIAL-MESENCHYMAL TRANSITION (EMT). EPIGENETIC MECHANISMS PLAY KEY ROLES IN EMT. DNA METHYLATION, MEDIATED BY DNA METHYLTRANSFERASES (DNMTS), IS AN EPIGENETIC MARKER THAT IS CRITICAL TO EMT. THE GOAL OF THIS STUDY WAS TO DETERMINE WHETHER DNMTS WERE INVOLVED IN TGF-BETA1-INDUCED EMT AND ELUCIDATE THE UNDERLYING MECHANISMS IN NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. GLOBAL DNA METHYLATION AND DNMT ACTIVITY WERE QUANTIFIED. DNMT EXPRESSION WAS MEASURED USING REAL-TIME PCR (QRT-PCR) IN HUMAN CRS TISSUES. MRNA AND PROTEIN LEVELS OF DNMTS, E-CADHERIN, VIMENTIN, ALPHA-SMA, AND FIBRONECTIN WERE DETERMINED USING RT-PCR AND WESTERN BLOTTING, RESPECTIVELY. DNMT1, DNMT3A, AND DNMT3B GENE EXPRESSION WERE KNOCKED DOWN USING SIRNA TRANSFECTION. MAPK PHOSPHORYLATION AND EMT-RELATED TRANSCRIPTION FACTOR LEVELS WERE DETERMINED USING WESTERN BLOTTING. SIGNALING PATHWAYS WERE ANALYZED USING SPECIFIC INHIBITORS OF MAPK. WE DEMONSTRATED THESE DATA IN PRIMARY NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. GLOBAL DNA METHYLATION, DNMT ACTIVITY, AND DNMT EXPRESSION INCREASED IN CRS TISSUES. DNMT EXPRESSION WAS POSITIVELY CORRELATED WITH LUND-MCKAY CT SCORES. TGF-BETA1 DOSE-DEPENDENTLY INDUCED DNMT EXPRESSION. FURTHER, 5-AZA INHIBITED TGF-BETA1-INDUCED DNMT, SNAIL, AND SLUG EXPRESSION RELATED TO EMT, AS WELL AS P38 AND JNK PHOSPHORYLATION IN A549 CELLS AND TGF-BETA1-INDUCED DNMT EXPRESSION AND EMT IN PRIMARY NASAL EPITHELIAL CELLS AND AIR-LIQUID INTERFACE CULTURES. TGF-BETA1-INDUCED DNMT EXPRESSION LEADS TO DNA METHYLATION AND EMT VIA P38, JNK, SNAIL, AND SLUG SIGNALING PATHWAYS. INHIBITION OF DNMT SUPPRESSED THE EMT PROCESS AND THEREFORE IS POTENTIALLY A CRS THERAPEUTIC STRATEGY. 2022 18 2435 31 EPIGENETIC SILENCING OF SFRP1 ACTIVATES THE CANONICAL WNT PATHWAY AND CONTRIBUTES TO INCREASED CELL GROWTH AND PROLIFERATION IN HEPATOCELLULAR CARCINOMA. THE WNT PATHWAY IS A KEY REGULATOR OF EMBRYONIC DEVELOPMENT AND STEM CELLS, AND ITS ABERRANT ACTIVATION IS ASSOCIATED WITH HUMAN MALIGNANCIES, MOST NOTABLY HEPATOCELLULAR CARCINOMA (HCC). EPIGENETIC DEREGULATION OF THE GENES ENCODING THE SECRETED FRIZZLED-RELATED PROTEINS (SFRPS), THE WNT SIGNALLING ANTAGONISTS, HAS BEEN LINKED WITH ABERRANT HYPERACTIVATION OF THE WNT SIGNALLING IN HCC CELLS; HOWEVER, THE PRECISE UNDERLYING MECHANISM REMAINS ELUSIVE. WE INVESTIGATED THE METHYLATION PROFILES OF WNT ANTAGONISTS IN LIVER SAMPLES OF DIFFERENT STAGES OF HCC DEVELOPMENT AND LIVER CANCER CELL LINES AND STUDIED THE FUNCTIONAL IMPACT OF ABERRANT EPIGENETIC SILENCING OF SFRPS ON THE CANONICAL WNT PATHWAY AND CELL VIABILITY. WE FOUND THAT THE SFRP1 GENE ENCODING THE SUBUNIT IS A FREQUENT TARGET OF ABERRANT DNA HYPERMETHYLATION AND SILENCING IN HCC TUMOURS, WHEREAS OTHER EXTRACELLULAR WNT ANTAGONISTS, WIF1 AND DKK3, EXHIBITED NO METHYLATION IN TUMOUR CELLS, CONSISTENT WITH THE NOTION THAT ABERRANT METHYLATION EVENTS IN CANCER CELLS ARE NON-RANDOMLY DISTRIBUTED AMONG THE GENES AND THAT THERE IS A STRONG PREFERENCE FOR HYPERMETHYLATION OF SPECIFIC GENES IN HCC. IN ADDITION, BY COMPARING SFRP1 METHYLATION STATUS IN HCC TUMOURS WITH NORMAL, CIRRHOTIC AND CHRONIC HEPATITIS LIVER TISSUES, WE IDENTIFIED SFRP1 GENE AS A POTENTIAL EARLY MARKER OF HCC. THE RESTORATION OF SFRP1 EXPRESSION IN CANCER CELLS BY ECTOPIC EXPRESSION INHIBITED WNT ACTIVITY ACCOMPANIED WITH DESTABILIZATION OF BETA-CATENIN AND DOWNREGULATION OF C-MYC AND CYCLIN D1, THE KNOWN DOWNSTREAM TARGETS OF WNT PATHWAY. IMPORTANTLY, RESTORING SFRP1 LEVELS IN CANCER CELLS INHIBITED CELL GROWTH AND INDUCED APOPTOTIC CELL DEATH. THIS STUDY SUPPORTS THE CRITICAL ROLE FOR SFRP1 SILENCING IN HEPATOCELLULAR CARCINOMA AND REINFORCES THE IMPORTANCE OF THE WNT ANTAGONISTS IN PREVENTING ONCOGENIC STABILIZATION OF BETA-CATENIN AND CHRONIC ACTIVATION OF THE CANONICAL WNT PATHWAY, SUGGESTING THAT SFRP1 MAY BE AN ATTRACTIVE TARGET FOR EARLY CANCER DETECTION AND THERAPEUTIC INTERVENTION. 2012 19 5083 23 PICKING A BONE WITH WISP1 (CCN4): NEW STRATEGIES AGAINST DEGENERATIVE JOINT DISEASE. AS THE WORLD'S POPULATION CONTINUES TO AGE, IT IS ESTIMATED THAT DEGENERATIVE JOINT DISEASE DISORDERS SUCH AS OSTEOARTHRITIS WILL IMPACT AT LEAST 130 MILLION INDIVIDUALS THROUGHOUT THE GLOBE BY THE YEAR 2050. ADVANCED AGE, OBESITY, GENETICS, GENDER, BONE DENSITY, TRAUMA, AND A POOR LEVEL OF PHYSICAL ACTIVITY CAN LEAD TO THE ONSET AND PROGRESSION OF OSTEOARTHRITIS. HOWEVER, FACTORS THAT LEAD TO DEGENERATIVE JOINT DISEASE AND INVOLVE GENDER, GENETICS, EPIGENETIC MECHANISMS, AND ADVANCED AGE ARE NOT WITHIN THE CONTROL OF AN INDIVIDUAL. FURTHERMORE, CURRENT THERAPIES INCLUDING PAIN MANAGEMENT, IMPROVED NUTRITION, AND REGULAR PROGRAMS FOR EXERCISE DO NOT LEAD TO THE RESOLUTION OF OSTEOARTHRITIS. AS A RESULT, NEW AVENUES FOR TARGETING THE TREATMENT OF OSTEOARTHRITIS ARE DESPERATELY NEEDED. WNT1 INDUCIBLE SIGNALING PATHWAY PROTEIN 1 (WISP1), A MATRICELLULAR PROTEIN AND A DOWNSTREAM TARGET OF THE WINGLESS PATHWAY WNT1, IS ONE SUCH TARGET TO CONSIDER THAT GOVERNS CELLULAR PROTECTION, STEM CELL PROLIFERATION, AND TISSUE REGENERATION IN A NUMBER OF DISORDERS INCLUDING BONE DEGENERATION. HOWEVER, INCREASED WISP1 EXPRESSION ALSO HAS BEEN ASSOCIATED WITH THE PROGRESSION OF OSTEOARTHRITIS. WISP1 HAS AN INTRICATE RELATIONSHIP WITH A NUMBER OF PROLIFERATIVE AND PROTECTIVE PATHWAYS THAT INCLUDE PHOSPHOINOSITIDE 3-KINASE (PI 3-K), PROTEIN KINASE B (AKT), NUCLEAR FACTOR KAPPA-LIGHT-CHAIN-ENHANCER OF ACTIVATED B CELLS (NF-KAPPAB), INTERLEUKIN -6 (IL-6), TRANSFORMING GROWTH FACTOR-BETA, MATRIX METALLOPROTEINASE, SMALL NON-CODING RIBONUCLEIC ACIDS (RNAS), SIRTUIN SILENT MATING TYPE INFORMATION REGULATION 2 HOMOLOG 1 (SACCHAROMYCES CEREVISIAE) (SIRT1), AND THE MECHANISTIC TARGET OF RAPAMYCIN (MTOR). TAKEN TOGETHER, THIS COMPLEX ASSOCIATION WISP1 HOLDS WITH THESE SIGNALING PATHWAYS NECESSITATES A FINE BIOLOGICAL REGULATION OF WISP1 ACTIVITY THAT CAN OFFSET THE PROGRESSION OF DEGENERATIVE JOINT DISEASE, BUT NOT LIMIT THE CELLULAR PROTECTIVE CAPABILITIES OF THE WISP1 PATHWAY. 2016 20 6235 26 THE M(6)A DEMETHYLASE FTO PROMOTES RENAL EPITHELIAL-MESENCHYMAL TRANSITION BY REDUCING THE M(6)A MODIFICATION OF LNCRNA GAS5. BACKGROUND: RENAL INTERSTITIAL FIBROSIS (RIF) IS THE MAIN PATHOLOGICAL CHANGE OF A VARIETY OF CHRONIC KIDNEY DISEASES (CKD). EPIGENETIC MODIFICATIONS OF FIBROSIS-PRONE GENES REGULATE RIF PROGRESSION. THIS STUDY AIMED TO INVESTIGATE LONG NON-CODING RNA (LNCRNA) N6-METHYLADENOSINE (M(6)A) MODIFICATION AND ITS ROLE IN REGULATING RIF PROGRESSION. METHODS: UNILATERAL URETERAL OCCLUSION (UUO) WAS EMPLOYED TO CONSTRUCT THE RIF IN VIVO MODEL; AND TGF-BETA1-TREATED HK-2 AND HKC-8 CELLS WERE USED FOR IN VITRO EXPERIMENTS. THE MRNA AND PROTEIN EXPRESSIONS WERE ASSESSED USING QRT-PCR AND WESTERN BLOT. THE PROLIFERATION AND MIGRATION WERE EVALUATED BY EDU ASSAY AND TRANSWELL ASSAY, RESPECTIVELY. IN ADDITION, LEVELS OF INFLAMMATORY CYTOKINES WERE DETERMINED BY ELISA ASSAY AND QRT-PCR. MOREOVER, LNCRNA GAS5 M(6)A LEVEL WAS DETECTED USING ME-RIP ASSAY. HE AND MASSON STAINING WERE EMPLOYED TO EVALUATE FIBROTIC LESIONS OF THE KIDNEY. RESULTS: FTO EXPRESSION WAS ELEVATED IN HK-2 AND HKC-8 CELLS AFTER TGF-BETA1 TREATMENT AND MOUSE KIDNEY TISSUE FOLLOWING UUO, AND LNCRNA GAS5 WAS DOWNREGULATED. LNCRNA GAS5 OVEREXPRESSION OR FTO SILENCING SUPPRESSED TGF-BETA1-INDUCED THE INCREASE OF EMT-RELATED PROTEINS (VIMENTIN, SNAIL AND N-CADHERIN) AND INFLAMMATORY CYTOKINES (IL-6, IL-1BETA AND TNF-ALPHA) LEVELS IN HK-2 CELLS. FTO SUPPRESSED LNCRNA GAS5 EXPRESSION BY REDUCING THE M6A MODIFICATION OF LNCRNA GAS5. ADDITIONALLY, FTO KNOCKDOWN COULD SUPPRESS EMT PROCESS AND INFLAMMATION RESPONSE INDUCED BY TGF-BETA1 AND UUO IN VITRO AND IN VIVO. AS EXPECTED, FTO KNOCKDOWN ABROGATED THE PROMOTION EFFECTS OF LNCRNA GAS5 SILENCING ON TGF-BETA1-INDUCED EMT PROCESS AND INFLAMMATION RESPONSE IN HK-2 AND HKC-8 CELLS. CONCLUSION: FTO PROMOTED EMT PROCESS AND INFLAMMATION RESPONSE THROUGH REDUCING THE M(6)A MODIFICATION OF LNCRNA GAS5. 2022