1 967 120 CHRONIC NICOTINE EXPOSURE AUGMENTS RENAL OXIDATIVE STRESS AND INJURY THROUGH TRANSCRIPTIONAL ACTIVATION OF P66SHC. BACKGROUND: CHRONIC NICOTINE (CH-NIC) EXPOSURE EXACERBATES ISCHEMIA/REPERFUSION (I/R)-INDUCED OXIDATIVE STRESS AND ACUTE KIDNEY INJURY (AKI), AND MITOCHONDRIAL PRODUCTION OF REACTIVE OXYGEN SPECIES (ROS) IN CULTURED RENAL PROXIMAL TUBULE CELLS (RPTCS). BECAUSE SER36-PHOSPHORYLATED P66SHC MODULATES MITOCHONDRIAL ROS PRODUCTION AND INJURY OF RPTCS, WE HYPOTHESIZED THAT CH-NIC EXACERBATES AKI BY INCREASING STRESS-INDUCED PHOSPHORYLATION OF P66SHC. METHODS: WE FIRST TESTED WHETHER CH-NIC AUGMENTS I/R-AKI-INDUCED EXPRESSION AND PHOSPHORYLATION OF P66SHC IN VIVO. WE THEN EXAMINED WHETHER KNOCKING DOWN P66SHC, OR IMPAIRING ITS SER36 PHOSPHORYLATION OR BINDING TO CYTOCHROME C, ALTERS THE EFFECTS OF CH-NIC ON OXIDATIVE STRESS (H(2)O(2))-INDUCED PRODUCTION OF ROS, MITOCHONDRIAL DEPOLARIZATION AND INJURY IN RPTCS IN VITRO. RESULTS: WE FOUND THAT CH-NIC INCREASED THE EXPRESSION OF P66SHC IN THE CONTROL AND ISCHEMIC KIDNEYS, BUT ONLY INCREASED ITS SER36 PHOSPHORYLATION AFTER RENAL I/R. KNOCKING DOWN P66SHC OR IMPAIRING PHOSPHORYLATION OF ITS SER36 RESIDUE, VIA THE S36A MUTATION (BUT NOT THE PHOSPHOMIMETIC S36D MUTATION), BLUNTED CH-NIC + H2O2-DEPENDENT ROS PRODUCTION, MITOCHONDRIAL DEPOLARIZATION AND INJURY IN RPTCS. ADDITIONALLY, CH-NIC + H2O2-DEPENDENT BINDING OF P66SHC TO MITOCHONDRIAL CYTOCHROME C WAS ATTENUATED BY S36A MUTATION OF P66SHC, AND IMPAIRING CYTOCHROME C BINDING (VIA W134F MUTATION) ABOLISHED ROS PRODUCTION, MITOCHONDRIAL DEPOLARIZATION AND INJURY, WHILE ECTOPIC OVEREXPRESSION OF P66SHC (WHICH MIMICS CH-NIC TREATMENT) AUGMENTED OXIDANT INJURY. WE DETERMINED THAT CH-NIC STIMULATES THE P66SHC PROMOTER THROUGH P53- AND EPIGENETIC MODIFICATION (PROMOTER HYPOMETHYLATION). CONCLUSIONS: CH-NIC WORSENS OXIDATIVE STRESS-DEPENDENT ACUTE RENAL INJURY BY INCREASING EXPRESSION AND CONSEQUENT OXIDATIVE STRESS-DEPENDENT SER36 PHOSPHORYLATION OF P66SHC. THUS, TARGETING THIS PATHWAY MAY HAVE THERAPEUTIC RELEVANCE IN PREVENTING/AMELIORATING TOBACCO-RELATED KIDNEY INJURY. 2013 2 2796 18 FBW7 MEDIATES SENESCENCE AND PULMONARY FIBROSIS THROUGH TELOMERE UNCAPPING. TISSUE STEM CELLS UNDERGO PREMATURE SENESCENCE UNDER STRESS, PROMOTING AGE-RELATED DISEASES; HOWEVER, THE ASSOCIATED MECHANISMS REMAIN UNCLEAR. HERE, WE REPORT THAT IN RESPONSE TO RADIATION, OXIDATIVE STRESS, OR BLEOMYCIN, THE E3 UBIQUITIN LIGASE FBW7 MEDIATES CELL SENESCENCE AND TISSUE FIBROSIS THROUGH TELOMERE UNCAPPING. FBW7 BINDING TO TELOMERE PROTECTION PROTEIN 1 (TPP1) FACILITATES TPP1 MULTISITE POLYUBIQUITINATION AND ACCELERATES DEGRADATION, TRIGGERING TELOMERE UNCAPPING AND DNA DAMAGE RESPONSE. OVEREXPRESSING TPP1 OR INHIBITING FBW7 BY GENETIC ABLATION, EPIGENETIC INTERFERENCE, OR PEPTIDOMIMETIC TELOMERE DYSFUNCTION INHIBITOR (TELODIN) REDUCES TELOMERE UNCAPPING AND SHORTENING, EXPANDING THE PULMONARY ALVEOLAR AEC2 STEM CELL POPULATION IN MICE. TELODIN, SYNTHESIZED FROM THE SEVENTH BETA STRAND BLADE OF FBW7 WD40 PROPELLER DOMAIN, INCREASES TPP1 STABILITY, LUNG RESPIRATORY FUNCTION, AND RESISTANCE TO SENESCENCE AND FIBROSIS IN ANIMALS CHRONICALLY EXPOSED TO ENVIRONMENTAL STRESS. OUR FINDINGS ELUCIDATE A PIVOTAL MECHANISM UNDERLYING STRESS-INDUCED PULMONARY EPITHELIAL STEM CELL SENESCENCE AND FIBROSIS, PROVIDING A FRAMEWORK FOR AGING-RELATED DISORDER INTERVENTIONS. 2020 3 2649 17 EPIGENOMIC, GENOMIC, AND TRANSCRIPTOMIC LANDSCAPE OF SCHWANNOMATOSIS. SCHWANNOMATOSIS (SWNTS) IS A GENETIC CANCER PREDISPOSITION SYNDROME THAT MANIFESTS AS MULTIPLE AND OFTEN PAINFUL NEURONAL TUMORS CALLED SCHWANNOMAS (SWNS). WHILE GERMLINE MUTATIONS IN SMARCB1 OR LZTR1, PLUS SOMATIC MUTATIONS IN NF2 AND LOSS OF HETEROZYGOSITY IN CHROMOSOME 22Q HAVE BEEN IDENTIFIED IN A SUBSET OF PATIENTS, LITTLE IS KNOWN ABOUT THE EPIGENOMIC AND GENOMIC ALTERATIONS THAT DRIVE SWNTS-RELATED SWNS (SWNTS-SWNS) IN A MAJORITY OF THE CASES. WE PERFORMED MULTIPLATFORM GENOMIC ANALYSIS AND ESTABLISHED THE MOLECULAR SIGNATURE OF SWNTS-SWNS. WE SHOW THAT SWNTS-SWNS HARBOR DISTINCT GENOMIC FEATURES RELATIVE TO THE HISTOLOGICALLY IDENTICAL NON-SYNDROMIC SPORADIC SWNS (NS-SWNS). WE DEMONSTRATE THE EXISTENCE OF FOUR DISTINCT DNA METHYLATION SUBGROUPS OF SWNTS-SWNS THAT ARE ASSOCIATED WITH SPECIFIC TRANSCRIPTIONAL PROGRAMS AND TUMOR LOCATION. WE SHOW SEVERAL NOVEL RECURRENT NON-22Q DELETIONS AND STRUCTURAL REARRANGEMENTS. WE DETECTED THE SH3PXD2A-HTRA1 GENE FUSION IN SWNTS-SWNS, WITH PREDOMINANCE IN LZTR1-MUTANT TUMORS. IN ADDITION, WE IDENTIFIED SPECIFIC GENETIC, EPIGENETIC, AND ACTIONABLE TRANSCRIPTIONAL PROGRAMS ASSOCIATED WITH PAINFUL SWNTS-SWNS INCLUDING PIGF, VEGF, MEK, AND MTOR PATHWAYS, WHICH MAY BE HARNESSED FOR MANAGEMENT OF THIS SYNDROME. 2021 4 4048 22 MAINTENANCE AND PHARMACOLOGIC TARGETING OF ROR1 PROTEIN LEVELS VIA UHRF1 IN T(1;19) PRE-B-ALL. EXPRESSION OF THE TRANSMEMBRANE PSEUDOKINASE ROR1 IS REQUIRED FOR SURVIVAL OF T(1;19)-PRE-B-CELL ACUTE LYMPHOBLASTIC LEUKEMIA (T(1;19) PRE-B-ALL), CHRONIC LYMPHOCYTIC LEUKEMIA, AND MANY SOLID TUMORS. HOWEVER, TARGETING ROR1 WITH SMALL-MOLECULES HAS BEEN CHALLENGING DUE TO THE ABSENCE OF ROR1 KINASE ACTIVITY. TO IDENTIFY GENES THAT REGULATE ROR1 EXPRESSION AND MAY, THEREFORE, SERVE AS SURROGATE DRUG TARGETS, WE EMPLOYED AN SIRNA SCREENING APPROACH AND DETERMINED THAT THE EPIGENETIC REGULATOR AND E3 UBIQUITIN LIGASE, UHRF1, IS REQUIRED FOR T(1;19) PRE-B-ALL CELL VIABILITY IN A ROR1-DEPENDENT MANNER. UPON UHRF1 SILENCING, ROR1 PROTEIN IS REDUCED WITHOUT ALTERING ROR1 MRNA, AND ECTOPICALLY EXPRESSED UHRF1 IS SUFFICIENT TO INCREASE ROR1 LEVELS. ADDITIONALLY, PROTEASOME INHIBITION RESCUES LOSS OF ROR1 PROTEIN AFTER UHRF1 SILENCING, SUGGESTING A ROLE FOR THE PROTEASOME IN THE UHRF1-ROR1 AXIS. FINALLY, WE SHOW THAT ROR1-POSITIVE CELLS ARE TWICE AS SENSITIVE TO THE UHRF1-TARGETING DRUG, NAPHTHAZARIN, AND UNDERGO INCREASED APOPTOSIS COMPARED TO ROR1-NEGATIVE CELLS. NAPHTHAZARIN ELICITS REDUCED EXPRESSION OF UHRF1 AND ROR1, AND COMBINATION OF NAPHTHAZARIN WITH INHIBITORS OF PRE-B CELL RECEPTOR SIGNALING RESULTS IN FURTHER REDUCTION OF CELL SURVIVAL COMPARED WITH EITHER INHIBITOR ALONE. THEREFORE, OUR WORK REVEALS A MECHANISM BY WHICH UHRF1 STABILIZES ROR1, SUGGESTING A POTENTIAL TARGETING STRATEGY TO INHIBIT ROR1 IN T(1;19) PRE-B-ALL AND OTHER MALIGNANCIES. 2018 5 3876 21 KDM4A-MEDIATED HISTONE DEMETHYLATION OF SLC7A11 INHIBITS CELL FERROPTOSIS IN OSTEOSARCOMA. OSTEOSARCOMA (OS) IS THE MOST COMMON TYPE OF BONE TUMOR THAT SERIOUSLY AFFECTS LIMB FUNCTION AND INDUCES GREAT PAIN IN PATIENTS. LUNG METASTASIS AND CHEMOTHERAPY RESISTANCE ARE TWO KEY ISSUES LEADING TO THE POOR PROGNOSIS OF OS PATIENTS, THEREFORE NEW TREATMENT TARGETS AND STRATEGIES ARE URGENTLY NEEDED. IN OUR STUDY, WE UNCOVERED THE ROLE OF HISTONE DEMETHYLASE KDM4A IN REGULATING OS CELL FERROPTOSIS AND TUMOR PROGRESSION. KDM4A WAS SIGNIFICANTLY UPREGULATED IN OS SPECIMENS AND HIGH KDM4A EXPRESSION WAS ASSOCIATED WITH POORER PROGNOSIS IN OS PATIENTS. OUR DATA INDICATED THAT TARGETING KDM4A SIGNIFICANTLY INCREASED OS CELL DEATH, ENHANCED CISPLATIN RESPONSE, AND ATTENUATED MIGRATION ABILITY IN VITRO. KDM4A DEPLETION DRAMATICALLY INHIBITED TUMOR PROGRESSION AND LUNG METASTASIS OF OS IN VIVO FURTHER EXPERIMENTS CONFIRMED THAT KDM4A KNOCKDOWN PROMOTED OS CELL FERROPTOSIS, A SPECIAL NON-APOPTOTIC FORM OF CELL DEATH. KDM4A REGULATES SLC7A11 TRANSCRIPTION AND OS CELL FERROPTOSIS BY CONTROLLING H3K9ME3 DEMETHYLATION IN THE PROMOTER REGION OF SLC7A11. OUR FINDINGS DEEPENED THE RECOGNITION OF EPIGENETIC REGULATORY MECHANISM IN OS TUMORIGENESIS, CHEMORESISTANCE, AND METASTASIS, SUGGESTING THAT KDM4A ACTIVITY MAY BE A POTENTIAL THERAPEUTIC TARGET FOR FUTURE OS TREATMENT. 2021 6 6463 19 TISSUE METHYLATION AND DEMETHYLATION INFLUENCE TRANSLESION SYNTHESIS DNA POLYMERASES (TLS) CONTRIBUTING TO THE GENESIS OF CHROMOSOMAL ABNORMALITIES IN MYELODYSPLASTIC SYNDROME. AIMS: DNA METHYLATION HAS ITS DISTRIBUTION INFLUENCED BY DNA DEMETHYLATION PROCESSES WITH THE CATALYTIC CONVERSION OF 5-METHYLCYTOSINE (5MC) INTO 5-HYDROXYMETHYLCYTOSINE (5HMC). MYELODYSPLASTIC SYNDROME (MDS) HAS BEEN ASSOCIATED WITH EPIGENETIC DYSREGULATION OF GENES RELATED TO DNA REPAIR SYSTEM, CHRONIC IMMUNE RESPONSE AND CELL CYCLE. METHODS: WE EVALUATED THE TISSUE DNA METHYLATION/HYDROXYMETHYLATION IN BONE MARROW TREPHINE BIOPSIES OF 73 PATIENTS WITH MDS, TRYING TO CORRELATE WITH THE MRNA EXPRESSION OF 21 GENES (POLH, POLL, REV3L, POLN, POLQ, POLI, POLK, IRF-1, IRF-2, IRF-3, IRF-4, IRF-5, IRF6, IRF-7, IRF-8,IRF-9, MAD2, CDC20, AURKA, AURKB AND TPX2). RESULTS: THE M-SCORE (5MC) WAS SIGNIFICANTLY HIGHER IN PATIENTS WITH CHROMOSOMAL ABNORMALITIES THAN PATIENTS WITH NORMAL KARYOTYPE (95% CI -27.127779 TO -2.368020; P=0.022). WE OBSERVED A HIGHER 5MC/5HMC RATIO IN PATIENTS CLASSIFIED AS HIGH-RISK SUBTYPES COMPARED WITH LOW-RISK SUBTYPES (95% CI -72.922115 TO -1.855662; P=0.040) AS WELL AS PATIENTS WITH HYPERCELLULAR BONE MARROW COMPARED WITH PATIENTS WITH NORMOCELLULAR/HYPOCELLULAR BONE MARROW (95% CI -69.189259 TO -0.511828; P=0.047) AND WITH THE PRESENCE OF DYSERYTHROPOIESIS (95% CI 17.077703 TO 51.331388; P=0.001). DNA POLS WITH TRANSLESION ACTIVITY ARE SIGNIFICANTLY INFLUENCED BY METHYLATION. AS 5MC IMMUNOEXPRESSION INCREASES, THE EXPRESSIONS OF POLH (R=-0.816; R(2) =0.665; P=0.000), POLQ (R=-0.790; R(2)=0.624; P=0.001), PCNA (R=-0.635; R(2)=0.403; P=0.020), POLK (R=-0.633; R(2)=0.400; P=0.036 AND REV1 (R=-0.578; R(2)=0.334; P=0.049) DECREASE. CONCLUSIONS: OUR RESULTS CONFIRM THAT THERE IS AN IMBALANCE IN THE DNA METHYLATION IN MDS, INFLUENCING THE DEVELOPMENT OF CHROMOSOMAL ABNORMALITIES WHICH MAY BE ASSOCIATED WITH THE LOW EXPRESSION OF DNA POLYMERASES WITH TRANSLESION SYNTHESIS POLYMERASES ACTIVITY. 2022 7 4443 19 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 8 850 18 CHILDREN WITH CHRONIC IMMUNE THROMBOCYTOPENIA EXHIBIT HIGH EXPRESSION OF HUMAN ENDOGENOUS RETROVIRUSES TRIM28 AND SETDB1. CHRONIC IMMUNE THROMBOCYTOPENIA (CITP) IS AN AUTOIMMUNE DISEASE WHOSE UNDERLYING BIOLOGIC MECHANISMS REMAIN ELUSIVE. HUMAN ENDOGENOUS RETROVIRUSES (HERVS) DERIVE FROM ANCESTRAL INFECTIONS AND CONSTITUTE ABOUT 8% OF OUR GENOME. A WEALTH OF CLINICAL AND EXPERIMENTAL STUDIES HIGHLIGHTS THEIR PIVOTAL PATHOGENETIC ROLE IN AUTOIMMUNE DISEASES. EPIGENETIC MECHANISMS, SUCH AS THOSE MODULATED BY TRIM28 AND SETDB1, ARE INVOLVED IN HERV ACTIVATION AND REGULATION OF IMMUNE RESPONSE. WE ASSESSED, THROUGH A POLYMERASE CHAIN REACTION REAL-TIME TAQMAN AMPLIFICATION ASSAY, THE TRANSCRIPTION LEVELS OF POL GENES OF HERV-H, HERV-K, AND HERV-W; ENV GENES OF SYNCYTIN (SYN)1, SYN2, AND HERV-W; AS WELL AS TRIM28 AND SETDB1 IN WHOLE BLOOD FROM 34 CHILDREN WITH CITP AND AGE-MATCHED HEALTHY CONTROLS (HC). THE TRANSCRIPTIONAL LEVELS OF ALL HERV SEQUENCES, WITH THE EXCEPTION OF HERV-W-ENV, WERE SIGNIFICANTLY ENHANCED IN CHILDREN WITH CITP AS COMPARED TO HC. PATIENTS ON ELTROMBOPAG TREATMENT EXHIBITED LOWER EXPRESSION OF SYN1, SYN2, AND HERV-W-ENV AS COMPARED TO UNTREATED PATIENTS. THE MRNA CONCENTRATIONS OF TRIM28 AND SETDB1 WERE SIGNIFICANTLY HIGHER AND WERE POSITIVELY CORRELATED WITH THOSE OF HERVS IN CITP PATIENTS. THE OVER-EXPRESSIONS OF HERVS AND TRIM28/SETDB1 AND THEIR POSITIVE CORRELATIONS IN PATIENTS WITH CITP ARE SUGGESTIVE CLUES OF THEIR CONTRIBUTION TO THE PATHOGENESIS OF THE DISEASE AND SUPPORT INNOVATIVE INTERVENTIONS TO INHIBIT HERV AND TRIM28/SETDB1 EXPRESSIONS IN PATIENTS UNRESPONSIVE TO STANDARD THERAPIES. 2023 9 1171 25 CONTRIBUTION OF MATURE HEPATOCYTES TO BILIARY REGENERATION IN RATS WITH ACUTE AND CHRONIC BILIARY INJURY. WHETHER HEPATOCYTES CAN CONVERT INTO BILIARY EPITHELIAL CELLS (BECS) DURING BILIARY INJURY IS MUCH DEBATED. TO TEST THIS CONCEPT, WE TRACED THE FATE OF GENETICALLY LABELED [DIPEPTIDYL PEPTIDASE IV (DPPIV)-POSITIVE] HEPATOCYTES IN HEPATOCYTE TRANSPLANTATION MODEL FOLLOWING ACUTE HEPATO-BILIARY INJURY INDUCED BY 4,4'-METHYLENE-DIANILINE (DAPM) AND D-GALACTOSAMINE (DAPM+D-GAL) AND IN DPPIV-CHIMERIC LIVER MODEL SUBJECTED TO ACUTE (DAPM+D-GAL) OR CHRONIC BILIARY INJURY CAUSED BY DAPM AND BILE DUCT LIGATION (DAPM+BDL). IN BOTH MODELS BEFORE BILIARY INJURY, BECS ARE UNIFORMLY DPPIV-DEFICIENT AND PROLIFERATION OF DPPIV-DEFICIENT HEPATOCYTES IS RESTRICTED BY RETRORSINE. WE FOUND THAT MATURE HEPATOCYTES UNDERWENT A STEPWISE CONVERSION INTO BECS AFTER BILIARY INJURY. IN THE HEPATOCYTE TRANSPLANTATION MODEL, DPPIV-POSITIVE HEPATOCYTES ENTRAPPED PERIPORTALLY PROLIFERATED, AND FORMED TWO-LAYERED PLATES ALONG PORTAL VEINS. WITHIN THE TWO-LAYERED PLATES, THE HEPATOCYTES GRADUALLY LOST THEIR HEPATOCYTIC IDENTITY, PROCEEDED THROUGH AN INTERMEDIATE STATE, ACQUIRED A BILIARY PHENOTYPE, AND SUBSEQUENTLY FORMED BILE DUCTS ALONG THE HILUM-TO-PERIPHERY AXIS. IN DPPIV-CHIMERIC LIVER MODEL, PERIPORTAL HEPATOCYTES EXPRESSING HEPATOCYTE NUCLEAR FACTOR-1BETA (HNF-1BETA) WERE EXCLUSIVELY DPPIV-POSITIVE AND WERE IN CONTINUITY TO DPPIV-POSITIVES BILE DUCTS. INHIBITION OF HEPATOCYTE PROLIFERATION BY ADDITIONAL DOSES OF RETRORSINE IN DPPIV-CHIMERIC LIVERS PREVENTED THE APPEARANCE OF DPPIV-POSITIVE BECS AFTER BILIARY INJURY. MOREOVER, ENRICHED DPPIV-POSITIVE BEC/HEPATIC OVAL CELL TRANSPLANTATION PRODUCED DPPIV-POSITIVE BECS OR BILE DUCTS IN UNEXPECTEDLY LOW FREQUENCY AND IN MID-LOBULAR REGIONS. THESE RESULTS TOGETHER SUGGEST THAT MATURE HEPATOCYTES BUT NOT CONTAMINATING BECS/HEPATIC OVAL CELLS ARE THE SOURCES OF PERIPORTAL DPPIV-POSITIVE BECS. WE CONCLUDE THAT MATURE HEPATOCYTES CONTRIBUTE TO BILIARY REGENERATION IN THE ENVIRONMENT OF ACUTE AND CHRONIC BILIARY INJURY THROUGH A DUCTAL PLATE CONFIGURATION WITHOUT THE NEED OF EXOGENOUSLY GENETIC OR EPIGENETIC MANIPULATION. 2015 10 5228 23 PRMT7 TARGETS OF FOXM1 CONTROLS ALVEOLAR MYOFIBROBLAST PROLIFERATION AND DIFFERENTIATION DURING ALVEOLOGENESIS. ALTHOUGH ABERRANT ALVEOLAR MYOFIBROBLASTS (AMYFS) PROLIFERATION AND DIFFERENTIATION ARE OFTEN ASSOCIATED WITH ABNORMAL LUNG DEVELOPMENT AND DISEASES, SUCH AS BRONCHOPULMONARY DYSPLASIA (BPD), CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD), AND IDIOPATHIC PULMONARY FIBROSIS (IPF), EPIGENETIC MECHANISMS REGULATING PROLIFERATION AND DIFFERENTIATION OF AMYFS REMAIN POORLY UNDERSTOOD. PROTEIN ARGININE METHYLTRANSFERASE 7 (PRMT7) IS THE ONLY REPORTED TYPE III ENZYME RESPONSIBLE FOR MONOMETHYLATION OF ARGININE RESIDUE ON BOTH HISTONE AND NONHISTONE SUBSTRATES. HERE WE PROVIDE EVIDENCE FOR PRMT7'S FUNCTION IN REGULATING AMYFS PROLIFERATION AND DIFFERENTIATION DURING LUNG ALVEOLOGENESIS. IN PRMT7-DEFICIENT MICE, WE FOUND REDUCED AMYFS PROLIFERATION AND DIFFERENTIATION, ABNORMAL ELASTIN DEPOSITION, AND FAILURE OF ALVEOLAR SEPTUM FORMATION. WE FURTHER SHOWN THAT ONCOGENE FORKHEAD BOX M1 (FOXM1) IS A DIRECT TARGET OF PRMT7 AND THAT PRMT7-CATALYZED MONOMETHYLATION AT HISTONE H4 ARGININE 3 (H4R3ME1) DIRECTLY ASSOCIATE WITH CHROMATIN OF FOXM1 TO ACTIVATE ITS TRANSCRIPTION, AND THEREBY REGULATE OF CELL CYCLE-RELATED GENES TO INHIBIT AMYFS PROLIFERATION AND DIFFERENTIATION. OVEREXPRESSION OF FOXM1 IN ISOLATED MYOFIBROBLASTS (MYFS) SIGNIFICANTLY RESCUED PRMT7-DEFICIENCY-INDUCED CELL PROLIFERATION AND DIFFERENTIATION DEFECTS. THUS, OUR RESULTS REVEAL A NOVEL EPIGENETIC MECHANISM THROUGH WHICH PRMT7-MEDIATED HISTONE ARGININE MONOMETHYLATION ACTIVATES FOXM1 TRANSCRIPTIONAL EXPRESSION TO REGULATE AMYFS PROLIFERATION AND DIFFERENTIATION DURING LUNG ALVEOLOGENESIS AND MAY REPRESENT A POTENTIAL TARGET FOR INTERVENTION IN PULMONARY DISEASES. 2021 11 1217 31 CREG PROTECTS FROM MYOCARDIAL ISCHEMIA/REPERFUSION INJURY BY REGULATING MYOCARDIAL AUTOPHAGY AND APOPTOSIS. AIMS: HUMAN CELLULAR REPRESSOR OF E1A-STIMULATED GENES (CREG) IS A SECRETED GLYCOPROTEIN THAT REGULATES TISSUE AND CELL HOMEOSTASIS AND HAS BEEN SHOWN TO ANTAGONIZE HEART FIBROSIS, WHICH INDICATES A POTENTIAL PROTECTIVE EFFECT OF CREG AGAINST CARDIOMYOCYTE CHRONIC DAMAGE. HOWEVER, LITTLE IS KNOWN ABOUT THE ROLE OF CREG IN MYOCARDIAL TISSUE ACUTE INJURY, IN THIS STUDY, WE AIMED TO INVESTIGATE THE ROLE OF CREG IN MYOCARDIAL ISCHEMIA/REPERFUSION (MI/R) INJURY AND CLARIFY THE MECHANISM OF ACTION. METHODS AND RESULTS: WILD-TYPE CREG (CREG(+/+)), HETEROZYGOUS CREG (CREG(+/-)) MICE AND MICE PRETREATED WITH INFUSION OF RECOMBINANT 0.3MG/KG.D CREG PROTEIN (RECREG(+/+)) WERE SUBJECTED TO 30MIN OF LEFT ASCENDING CORONARY ISCHEMIA AND 24H OF REPERFUSION. EVAN'S BLUE-TRIPHENYL- TETRAZOLIUM CHLORIDE (TTC) SOLUTION AND ECHOCARDIOGRAPHY ANALYSIS WERE USED TO EVALUATE THE EFFECTS OF CREG ON MI/R MICE. THE UNDERLYING MECHANISMS WERE FURTHER DETERMINED BY CULTURED MYOCARDIAL CELLS IN VITRO. OUR FINDINGS REVEALED THAT THE LEVEL OF CREG PROTEIN IN MOUSE HEARTS WAS SIGNIFICANTLY DECREASED AFTER MICE WERE SUBJECTED TO MI/R. MOREOVER, CREG(+/-) MICE HAD LARGER INFARCTION SIZE 2H AFTER REPERFUSION AND WORSE CARDIAC FUNCTION 28DAYS AFTER MI/R INJURY COMPARED TO THAT IN CREG(+/+) MICE. HOWEVER, RECREG(+/+) MICE COULD MAINTAIN CREG AT A HIGH LEVEL EVEN AFTER MI/R INJURY, AND MITIGATED INFARCTION SIZE AND IMPROVED CARDIAC FUNCTION SIGNIFICANTLY. IN CREG(+/-) MICE, MYOCARDIAL AUTOPHAGY WAS DYSFUNCTIONAL CHARACTERIZED BY ACCUMULATION OF LC3A AND P62, WHILE APOPTOTIC CELL NUMBER INCREASE WAS DETECTED BY CLEAVED CASPASE-3 BLOTTING AND TUNEL STAINING. CONVERSELY, DECREASED APOPTOSIS AND ACTIVATED AUTOPHAGY WERE DETECTED IN RECREG(+/+) MICE. FURTHERMORE, CHLOROQUINE, A KIND OF AUTOPHAGY BLOCKER, WAS USED TO DEMONSTRATE RECOMBINANT CREG PROTECTED CARDIOMYOCYTES AGAINST APOPTOSIS MEDIATED BY ACTIVATING AUTOPHAGY BOTH IN VIVO AND IN VITRO. FINALLY, WE FOUND CREG WAS INVOLVED INTO LYSOSOMAL PROTEIN TRANSFER AND IMPROVE CELLULAR AUTOPHAGY. CONCLUSION: CREG PROTECTS HEART AGAINST MI/R INJURY-INDUCED CARDIOMYOCYTES APOPTOSIS BY ACTIVATING LYSOSOMAL AUTOPHAGY. THIS ARTICLE IS PART OF A SPECIAL ISSUE ENTITLED: GENETIC AND EPIGENETIC CONTROL OF HEART FAILURE - EDITED BY JUN REN AND MEGAN YINGMEI ZHANG. 2017 12 5298 23 PROTEIN ARGININE METHYLTRANSFERASE 5 SUPPRESSES THE TRANSCRIPTION OF THE RB FAMILY OF TUMOR SUPPRESSORS IN LEUKEMIA AND LYMPHOMA CELLS. THE PROPER EPIGENETIC MODIFICATION OF CHROMATIN BY PROTEIN ARGININE METHYLTRANSFERASES (PRMTS) IS CRUCIAL FOR NORMAL CELL GROWTH AND HEALTH. THE HUMAN SWI/SNF-ASSOCIATED PRMT5 IS INVOLVED IN THE TRANSCRIPTIONAL REPRESSION OF TARGET GENES BY DIRECTLY METHYLATING H3R8 AND H4R3. TO FURTHER UNDERSTAND THE IMPACT OF PRMT5-MEDIATED HISTONE METHYLATION ON CANCER, WE ANALYZED ITS EXPRESSION IN NORMAL AND TRANSFORMED HUMAN B LYMPHOCYTES. OUR FINDINGS REVEAL THAT PRMT5 PROTEIN LEVELS ARE ENHANCED IN VARIOUS HUMAN LYMPHOID CANCER CELLS, INCLUDING TRANSFORMED CHRONIC LYMPHOCYTIC LEUKEMIA (B-CLL) CELL LINES. PRMT5 OVEREXPRESSION IS CAUSED BY THE ALTERED EXPRESSION OF THE PRMT5-SPECIFIC MICRORNAS 19A, 25, 32, 92, 92B, AND 96 AND RESULTS IN THE INCREASED GLOBAL SYMMETRIC METHYLATION OF H3R8 AND H4R3. AN EVALUATION OF BOTH EPIGENETIC MARKS AT PRMT5 TARGET GENES SUCH AS RB1 (P105), RBL1 (P107), AND RBL2 (P130) SHOWED THAT PROMOTERS H3R8 AND H4R3 ARE HYPERMETHYLATED, WHICH IN TURN TRIGGERS POCKET PROTEIN TRANSCRIPTIONAL REPRESSION. FURTHERMORE, REDUCING PRMT5 EXPRESSION IN WAC3CD5 B-CLL CELLS ABOLISHES H3R8 AND H4R3 HYPERMETHYLATION, RESTORES RBL2 EXPRESSION, AND INHIBITS CANCER CELL PROLIFERATION. THESE RESULTS INDICATE THAT PRMT5 OVEREXPRESSION EPIGENETICALLY ALTERS THE TRANSCRIPTION OF KEY TUMOR SUPPRESSOR GENES AND SUGGEST A CAUSAL ROLE OF THE ELEVATED SYMMETRIC METHYLATION OF H3R8 AND H4R3 AT THE RBL2 PROMOTER IN TRANSFORMED B-LYMPHOCYTE PATHOLOGY. 2008 13 2640 23 EPIGENOMIC AND TRANSCRIPTIONAL PROFILING IDENTIFIES IMPAIRED GLYOXYLATE DETOXIFICATION IN NAFLD AS A RISK FACTOR FOR HYPEROXALURIA. EPIGENETIC MODIFICATIONS (E.G. DNA METHYLATION) IN NAFLD AND THEIR CONTRIBUTION TO DISEASE PROGRESSION AND EXTRAHEPATIC COMPLICATIONS ARE POORLY EXPLORED. HERE, WE USE AN INTEGRATED EPIGENOME AND TRANSCRIPTOME ANALYSIS OF MOUSE NAFLD HEPATOCYTES AND IDENTIFY ALTERATIONS IN GLYOXYLATE METABOLISM, A PATHWAY RELEVANT IN KIDNEY DAMAGE VIA OXALATE RELEASE-A HARMFUL WASTE PRODUCT AND KIDNEY STONE-PROMOTING FACTOR. DOWNREGULATION AND HYPERMETHYLATION OF ALANINE-GLYOXYLATE AMINOTRANSFERASE (AGXT), WHICH DETOXIFIES GLYOXYLATE, PREVENTING EXCESSIVE OXALATE ACCUMULATION, IS ACCOMPANIED BY INCREASED OXALATE FORMATION AFTER METABOLISM OF THE PRECURSOR HYDROXYPROLINE. VIRAL-MEDIATED AGXT TRANSFER OR INHIBITING HYDROXYPROLINE CATABOLISM RESCUES EXCESSIVE OXALATE RELEASE. IN HUMAN STEATOTIC HEPATOCYTES, AGXT IS ALSO DOWNREGULATED AND HYPERMETHYLATED, AND IN NAFLD ADOLESCENTS, STEATOSIS SEVERITY CORRELATES WITH URINARY OXALATE EXCRETION. THUS, THIS WORK IDENTIFIES A REDUCED CAPACITY OF THE STEATOTIC LIVER TO DETOXIFY GLYOXYLATE, TRIGGERING ELEVATED OXALATE, AND PROVIDES A MECHANISTIC EXPLANATION FOR THE INCREASED RISK OF KIDNEY STONES AND CHRONIC KIDNEY DISEASE IN NAFLD PATIENTS. 2021 14 3236 23 HEN EGG LYSOZYME ALLEVIATES STATIC MECHANICAL PAIN VIA NRF1-PARKIN-TACAN SIGNALING AXIS IN SENSORY NEURONS. MECHANICAL ALLODYNIA IMPINGES ON THE LIFE QUALITY OF PATIENTS. HEN EGG LYSOZYME (HEL) IS A SUBSTANCE EXTRACTED FROM EGGS THAT IS COMMONLY USED TO INHIBIT BACTERIAL ACTIVITY. THE ROLE OF HEL IN REGULATING AND TREATING PAIN IS UNCLEAR. HERE, WE FIND THAT HEL SELECTIVELY ATTENUATES STATIC MECHANICAL ALLODYNIA OF MICE INDUCED BY COMPLETE FREUND'S ADJUVANT (CFA), SPINAL NERVE LIGATION (SNL) AND CHEMOTHERAPEUTIC AGENT. RNA-SEQ SCREENING REVEALS THAT CFA SIGNIFICANTLY REDUCES THE EXPRESSION OF PARKIN IN DORSAL ROOT GANGLION (DRG) NEURONS OF MICE, WHILE PRE-ADMINISTRATION OF HEL INCREASES THE EXPRESSION OF PARKIN AND REMITS THE STATIC MECHANICAL ALLODYNIA INDUCED BY PARKIN-SIRNA. MOREOVER, HEL INCREASES THE INTERACTION BETWEEN NUCLEAR RESPIRATORY FACTOR 1 (NRF1) AND HISTONE ACETYLTRANSFERASE P300 AND THEN ENHANCES THE NRF1 MEDIATED HISTONE ACETYLATION IN PRKN PROMOTER REGION IN DRGS OF MICE. FURTHER, PARKIN INTERACTS WITH MECHANOTRANSDUCING ION CHANNEL TACAN (TMEM120A) AND KNOCKDOWN OF PARKIN SIGNIFICANTLY INCREASES THE MEMBRANE TRAFFICKING OF TACAN IN SENSORY NEURONS OF MICE. WHILE PRE-ADMINISTRATION OF HEL INHIBITS THE INCREASED MEMBRANE TRAFFICKING OF TACAN IN SENSORY NEURONS OF MICE INDUCED BY PARKIN-SIRNA. IN ADDITION, PRE-GIVEN OF HEL ALSO SIGNIFICANTLY ATTENUATES THE STATIC MECHANICAL ALLODYNIA INDUCED BY OVEREXPRESSION OF TACAN IN MICE, AND THE EFFECT OF HEL CAN BE BLOCKED BY PARKIN-SIRNA. THIS INDICATES THAT HEL INCREASES THE EXPRESSION OF PARKIN THROUGH EPIGENETIC MECHANISMS AND THEN DECREASES TACAN MEMBRANE TRAFFICKING IN SENSORY NEURONS TO RELIEVE STATIC MECHANICAL HYPERSENSITIVITY. THEREFORE, WE REVEAL A NOVEL FUNCTION OF HEL, WHICH IS A POTENTIAL SUBSTANCE FOR THE TREATMENT OF STATIC MECHANICAL PAIN. 2022 15 16 27 4CRNA NEAT1 SPONGE ADSORPTION OF MIR-378 MODULATES ACTIVITY OF LIPOPOLYSACCHARIDE-TREATED ARTICULAR CHONDROCYTES AND INFLUENCES THE PATHOLOGICAL DEVELOPMENT OF OSTEOARTHRITIS. CONTEXT: OSTEOARTHRITIS (OA) IS A CHRONIC JOINT DISEASE THAT CAN EVENTUALLY LEAD TO DEGENERATION, FIBROSIS, FRACTURES, AND DEFECTS OF THE ARTICULAR CARTILAGE. LONG NON-CODING RNA (LNCRNA) IS A KEY SUBSTANCE IN MANY PROCESSES, SUCH AS EPIGENETIC REGULATION AND CELL-CYCLE AND CELL-DIFFERENTIATION MODULATION, AND ITS RELATIONSHIP WITH OA HAS BEEN REPEATEDLY VERIFIED. OBJECTIVE: THE STUDY INTENDED TO CLARIFY THE INFLUENCE OF LNCRNA NUCLEAR ENRICHED ABUNDANT TRANSCRIPT 1 (NEAT1), LNCRNA NEAT1, ON LIPOPOLYSACCHARIDE (LPS)-INDUCED OA CHONDROCYTES THROUGH SPONGE ADSORPTION OF MICRORNA-378 (MIR-378) AND TO PROVIDE NOVEL INSIGHTS INTO FUTURE DIAGNOSIS AND TREATMENT OF OA. DESIGN: THE RESEARCH TEAM PERFORMED AN ANIMAL STUDY. SETTING: THE STUDY TOOK PLACE IN THE DEPARTMENT OF REHABILITATION MEDICINE AT LINYI PEOPLE'S HOSPITAL IN LINYI, SHANGDONG, CHINA. ANIMALS: THE STUDY'S ANIMALS WERE 10 SPRAGUE DAWLEY (SD) RATS, 3-5 DAYS OLD AND 10-15 G IN WEIGHT, OF THE SPECIFIC-PATHOGEN-FREE (SPF) GRADE. INTERVENTION: THE RAT CHONDROCYTES FOR THE POSITIVE CONTROL GROUP (THE MODEL GROUP) WERE TREATED WITH 500 NG/ML OF LPS TO INDUCE OA. CHONDROCYTES TREATED WITH THE SAME AMOUNT OF NORMAL SALINE WERE USED AS THE NEGATIVE CONTROL GROUP. THE CHONDROCYTES OF THE LPS-INDUCED RATS WERE INTO SIX GROUPS: (1) A POSITIVE CONTROL GROUP TRANSFECTED WITH NEAT1-INTERFERING RNA, THE SH-NEAT1 GROUP; (2) A NEGATIVE CONTROL GROUP NOT TRANSFECTED WITH NEAT1-INTERFERING RNA, THE NEAT1 EMPTY VECTOR (NC-NEAT1) GROUP; (3) AN INTERVENTION GROUP CO-TRANSFECTED WITH NEAT1 INTERFERING RNA AND THE MIR-378 INHIBITOR SEQUENCE (INH-MIR-378 THE SH-NEAT1+ INH-MIR-378 GROUP; (4) A NEGATIVE CONTROL GROUP TRANSFECTED WITH NEAT1 INTERFERING RNA BUT NOT TRANSFECTED WITH THE MIR-378 INHIBITOR SEQUENCE, THE SH-NEAT1+ MIR-378 NEGATIVE CONTROL (NC-MIR-378) GROUP; (5) A NEGATIVE CONTROL GROUP TRANSFECTED WITH THE MIR-378 INHIBITOR SEQUENCE BUT NOT TRANSFECTED WITH NEAT1 INTERFERING RNA, THE NEAT1 EMPTY VECTOR (NC-NEAT1) + INH-MIR-378 GROUP; (6) A NEGATIVE CONTROL GROUP NOT TRANSFECTED WITH EITHER NEAT1 INTERFERING RNA OR THE MIR-378 INHIBITOR SEQUENCE, THE NC-NEAT1 + NC-MIR-378 GROUP. OUTCOME MEASURES: AN OA-CHONDROCYTE MODEL WAS INDUCED BY LPS AND MEASUREMENTS OF NEAT1 AND MIR-378 EXPRESSION WERE MADE BY REAL-TIME QUANTITATIVE REVERSE TRANSCRIPTION (QRT)- POLYMERASE CHAIN REACTION (PCR). THEN, SMALL NEAT1-INTERFERING RNA (SH-NEAT1), EMPTY VECTOR NEAT1 (NC-NEAT1), INHIBITOR-SEQUENCE-MIR-378 (INH-MIR-378), AND NEGATIVE-CONTROL-MIR-378 (NC-MIR-378) WERE TRANSFECTED INTO CELLS, AND CELL VIABILITY AND APOPTOSIS RATE WERE MEASURED. FINALLY, THE STUDY VERIFIED THE RELATIONSHIP BETWEEN NEAT1 AND MIR-378. RESULTS: COMPARED TO THE CONTROL GROUP, NEAT1 WAS SIGNIFICANTLY ELEVATED IN THE MODEL GROUP, AND ITS MIR-378 WAS SIGNIFICANTLY DECREASED. SILENCING NEAT1 CAN ENHANCE OA-CHONDROCYTE ACTIVITY AND DECREASE APOPTOSIS. WHEN NEAT1 AND MIR-378 WERE INHIBITED TOGETHER, AS SHOWN FORT THE NC-NEAT1 + NC-MIR-378 GROUP, NEAT1 EXPRESSION, AS WELL AS THE MULTIPLICATION AND APOPTOSIS ABILITY OF THE OA-MODEL CELLS, WERE THE SAME AS THOSE OF CELLS TRANSFECTED WITH AN EMPTY VECTOR, THE NC-NEAT1 GROUP. ALSO, THE NEAT1 + NC-MIR-378 GROUP'S CELL ACTIVITY WAS LOWER THAN THAT OF THE SH-NEAT1+NC-MIR-378 GROUP BUT HIGHER THAN THAT OF THE NC-NEAT1 + INH-MIR-378 GROUP. FINALLY, HIGHER FLUORESCENCE ACTIVITY OCCURRED FOR NEAT1-MUTANT TYPE (MUT) TRANSFECTED WITH INH-MIR-378. CONCLUSIONS: NEAT1, WHICH IS HIGHLY EXPRESSED IN OA, MEDIATES LPS-INDUCED OA-CHONDROCYTE ACTIVITY THROUGH SPONGE ADSORPTION OF MIR-378. 2022 16 3303 15 HIGH-FREQUENCY P16(INK) (4A) PROMOTER METHYLATION IS ASSOCIATED WITH HISTONE METHYLTRANSFERASE SETDB1 EXPRESSION IN SPORADIC CUTANEOUS MELANOMA. EPIGENETIC MECHANISMS PARTICIPATE IN MELANOMA DEVELOPMENT AND PROGRESSION. THE EFFECT OF HISTONE MODIFICATIONS AND THEIR CATALYSING ENZYMES OVER EUCHROMATIC PROMOTER DNA METHYLATION IN MELANOMA REMAINS UNCLEAR. THIS STUDY INVESTIGATED THE POTENTIAL ASSOCIATION OF P16(INK) (4A) PROMOTER METHYLATION WITH HISTONE METHYLTRANSFERASE SETDB1 EXPRESSION IN GREEK PATIENTS WITH SPORADIC MELANOMA AND THEIR CORRELATION WITH CLINICOPATHOLOGICAL CHARACTERISTICS. PROMOTER METHYLATION WAS DETECTED BY METHYLATION-SPECIFIC PCR IN 100 PERIPHERAL BLOOD SAMPLES AND 58 MELANOMA TISSUES FROM THE SAME PATIENTS. CELL PROLIFERATION (KI-67 INDEX), P16(INK) (4A) AND SETDB1 EXPRESSION WERE EVALUATED BY IMMUNOHISTOCHEMISTRY. HIGH-FREQUENCY PROMOTER METHYLATION (25.86%) WAS OBSERVED IN TISSUE SAMPLES AND CORRELATED WITH INCREASED CELL PROLIFERATION (P = 0.0514). P16(INK) (4A) PROMOTER METHYLATION WAS HIGHER IN VERTICAL GROWTH-PHASE (60%) MELANOMAS THAN IN RADIAL (40%, P = 0.063) AND THOSE DISPLAYING EPIDERMAL INVOLVEMENT (P = 0.046). IMPORTANTLY, P16(INK) (4A) METHYLATION CORRELATED WITH INCREASED MELANOMA THICKNESS ACCORDING TO BRESLOW INDEX (P = 0.0495) AND MARGINALLY WITH INCREASED CLARK LEVEL (I/II VS III/IV/V, P = 0.070). LOW (1-30%) P16(INK) (4A) EXPRESSION WAS DETECTED AT THE MAJORITY (19 OF 54) OF MELANOMA CASES (35.19%), BEING MARGINALLY CORRELATED WITH TUMOR LYMPHOCYTIC INFILTRATION (P = 0.078). SETDB1 NUCLEAR IMMUNOREACTIVITY WAS OBSERVED IN 47 OF 57 (82.46%) CASES, WHEREAS 27 OF 57 (47.37%) SHOWED CYTOPLASMIC IMMUNOEXPRESSION. CYTOPLASMIC SETDB1 EXPRESSION CORRELATED WITH HIGHER FREQUENCY OF P16(INK) (4A) METHYLATION AND P16(INK) (4A) EXPRESSION (P = 0.033, P = 0.011, RESPECTIVELY). INCREASED NUCLEAR SETDB1 LEVELS WERE ASSOCIATED WITH HIGHER MITOTIC COUNT (0-5/MM(2) VS >5/MM(2) , P = 0.0869), ADVANCED CLARK LEVEL (III-V, P = 0.0380), EPIDERMAL INVOLVEMENT (P = 0.0331) AND THE NON-CHRONIC SUN EXPOSURE-ASSOCIATED MELANOMA TYPE (P = 0.0664). OUR DATA DEMONSTRATE FOR THE FIRST TIME THE ASSOCIATION OF HISTONE METHYLTRANSFERASE SETDB1 WITH FREQUENT METHYLATION OF THE EUCHROMATIC P16(INK) (4A) PROMOTER AND SEVERAL PROGNOSTIC PARAMETERS IN MELANOMAS. 2014 17 6072 20 THE DNA METHYLOME OF HUMAN VASCULAR ENDOTHELIUM AND ITS USE IN LIQUID BIOPSIES. BACKGROUND: VASCULAR ENDOTHELIAL CELLS (VECS) ARE AN ESSENTIAL COMPONENT OF EACH TISSUE, CONTRIBUTE TO MULTIPLE PATHOLOGIES, AND ARE TARGETED BY IMPORTANT DRUGS. YET, THERE IS A SHORTAGE OF BIOMARKERS TO ASSESS VEC TURNOVER. METHODS: TO DEVELOP DNA METHYLATION-BASED LIQUID BIOPSIES FOR VECS, WE DETERMINED THE METHYLOME OF VECS ISOLATED FROM FRESHLY DISSOCIATED HUMAN TISSUES. FINDINGS: A COMPARISON WITH A HUMAN CELL-TYPE METHYLOME ATLAS YIELDED THOUSANDS OF LOCI THAT ARE UNIQUELY UNMETHYLATED IN VECS. THESE SITES ARE TYPICALLY GENE ENHANCERS, OFTEN RESIDING ADJACENT TO VEC-SPECIFIC GENES. WE ALSO IDENTIFIED HUNDREDS OF GENOMIC LOCI THAT ARE DIFFERENTIALLY METHYLATED IN ORGANOTYPIC VECS, INDICATING THAT VECS FEEDING SPECIFIC ORGANS ARE DISTINCT CELL TYPES WITH A STABLE EPIGENETIC IDENTITY. WE ESTABLISHED UNIVERSAL AND LUNG-SPECIFIC VEC MARKERS AND EVALUATED THEIR PRESENCE IN CIRCULATING CELL-FREE DNA (CFDNA). NEARLY 2.5% OF CFDNA IN THE PLASMA OF HEALTHY INDIVIDUALS ORIGINATES FROM VECS. SEPSIS, GRAFT VERSUS HOST DISEASE, AND CARDIAC CATHETERIZATION ARE ASSOCIATED WITH ELEVATED LEVELS OF VEC-DERIVED CFDNA, INDICATIVE OF VASCULAR DAMAGE. LUNG-SPECIFIC VEC CFDNA IS SELECTIVELY ELEVATED IN PATIENTS WITH CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) OR LUNG CANCER, REVEALING TISSUE-SPECIFIC VASCULAR TURNOVER. CONCLUSIONS: VEC CFDNA BIOMARKERS INFORM VASCULAR DYNAMICS IN HEALTH AND DISEASE, POTENTIALLY CONTRIBUTING TO EARLY DIAGNOSIS AND MONITORING OF PATHOLOGIES, AND ASSESSMENT OF DRUG ACTIVITY. FUNDING: THIS WORK WAS SUPPORTED BY THE BEUTLER RESEARCH PROGRAM, HELMSLEY CHARITABLE TRUST, JDRF, GRAIL AND THE DON FOUNDATION (TO Y.D.). Y.D HOLDS THE WALTER & GRETA STIEL CHAIR IN HEART STUDIES. B.G., R.S., J.M., D.N., T.K., AND Y.D. FILED PATENTS ON CFDNA ANALYSIS. 2023 18 1274 26 DACH1 PROTECTS PODOCYTES FROM EXPERIMENTAL DIABETIC INJURY AND MODULATES PTIP-H3K4ME3 ACTIVITY. DACHSHUND HOMOLOG 1 (DACH1), A KEY CELL-FATE DETERMINANT, REGULATES TRANSCRIPTION BY DNA SEQUENCE-SPECIFIC BINDING. WE IDENTIFIED DIMINISHED DACH1 EXPRESSION IN A LARGE-SCALE SCREEN FOR MUTATIONS THAT CONVERT INJURY-RESISTANT PODOCYTES INTO INJURY-SUSCEPTIBLE PODOCYTES. IN DIABETIC KIDNEY DISEASE (DKD) PATIENTS, PODOCYTE DACH1 EXPRESSION LEVELS ARE DIMINISHED, A CONDITION THAT STRONGLY CORRELATES WITH POOR CLINICAL OUTCOMES. GLOBAL DACH1 KO MICE MANIFEST RENAL HYPOPLASIA AND DIE PERINATALLY. PODOCYTE-SPECIFIC DACH1 KO MICE, HOWEVER, MAINTAIN NORMAL GLOMERULAR ARCHITECTURE AT BASELINE, BUT RAPIDLY EXHIBIT PODOCYTE INJURY AFTER DIABETES ONSET. FURTHERMORE, PODOCYTE-SPECIFIC AUGMENTATION OF DACH1 EXPRESSION IN MICE PROTECTS FROM DKD. COMBINED RNA SEQUENCING AND IN SILICO PROMOTER ANALYSIS REVEAL CONVERSELY OVERLAPPING GLOMERULAR TRANSCRIPTOMIC SIGNATURES BETWEEN PODOCYTE-SPECIFIC DACH1 AND PAX TRANSACTIVATION-DOMAIN INTERACTING PROTEIN (PTIP) KO MICE, WITH UPREGULATED GENES POSSESSING HIGHER-THAN-EXPECTED NUMBERS OF PROMOTER DACH1-BINDING SITES. PTIP, AN ESSENTIAL COMPONENT OF THE ACTIVATING HISTONE H3 LYSINE 4 TRIMETHYLATION (H3K4ME3) COMPLEX, INTERACTS WITH DACH1 AND IS RECRUITED BY DACH1 TO ITS PROMOTER-BINDING SITES. DACH1-PTIP RECRUITMENT REPRESSES TRANSCRIPTION AND REDUCES PROMOTER H3K4ME3 LEVELS. DACH1 KNOCKDOWN IN PODOCYTES COMBINED WITH HYPERGLYCEMIA TRIGGERS TARGET GENE UPREGULATION AND INCREASES PROMOTER H3K4ME3. THESE FINDINGS REVEAL THAT IN DKD, DIMINISHED DACH1 EXPRESSION ENHANCES PODOCYTE INJURY VULNERABILITY VIA EPIGENETIC DEREPRESSION OF ITS TARGET GENES. 2021 19 5677 29 SHORT AIP1 (ASK1-INTERACTING PROTEIN-1) ISOFORM LOCALIZES TO THE MITOCHONDRIA AND PROMOTES VASCULAR DYSFUNCTION. OBJECTIVE: VASCULAR ENDOTHELIAL CELLS (ECS) NORMALLY MAINTAIN VASCULAR HOMEOSTASIS AND ARE REGULATED BY PROINFLAMMATORY CYTOKINES AND REACTIVE OXYGEN SPECIES. A HUMAN GENOME-WIDE ASSOCIATION STUDY IDENTIFIED THAT AIP1 (ASK1 [APOPTOSIS SIGNAL-REGULATING KINASE 1]-INTERACTING PROTEIN-1; ALSO IDENTIFIED AS DAB2IP) GENE VARIANTS CONFER SUSCEPTIBILITY TO CARDIOVASCULAR DISEASE, BUT THE UNDERLYING MECHANISM IS UNKNOWN. APPROACH AND RESULTS: WE DETECTED A NORMAL AIP1 FORM (NAMED AIP1A) IN THE HEALTHY AORTA, BUT A SHORTER FORM OF AIP1 (NAMED AIP1B) WAS FOUND IN DISEASED AORTAE THAT CONTAINED ATHEROSCLEROTIC PLAQUES AND GRAFT ARTERIOSCLEROSIS. AIP1B TRANSCRIPTION IN RESTING ECS WAS SUPPRESSED THROUGH EPIGENETIC INHIBITION BY RIF1 (RAP1 [RAS-RELATED PROTEIN 1]-INTERACTING FACTOR 1)/H3K9 (HISTONE H3 LYSINE 9) METHYLTRANSFERASE-MEDIATED H3K9 TRIMETHYLATION, AND THIS INHIBITION WAS RELEASED BY PROINFLAMMATORY CYTOKINES. AIP1A, BUT NOT AIP1B, WAS DOWNREGULATED BY PROTEOLYTIC DEGRADATION THROUGH A SMURF1 (SMAD [SUPPRESSOR OF MOTHERS AGAINST DECAPENTAPLEGIC MISCELLANEOUS] UBIQUITYLATION REGULATORY FACTOR 1)-DEPENDENT PATHWAY IN ECS UNDER INFLAMMATION. THEREFORE, AIP1B WAS THE MAJOR FORM PRESENT DURING INFLAMMATORY CONDITIONS. AIP1B, WHICH LACKS THE N-TERMINAL PLECKSTRIN HOMOLOGY DOMAIN OF AIP1A, LOCALIZED TO THE MITOCHONDRIA AND AUGMENTED TNFALPHA (TUMOR NECROSIS FACTOR ALPHA)-INDUCED MITOCHONDRIAL REACTIVE OXYGEN SPECIES GENERATION AND EC ACTIVATION. AIP1B-ECTG (EC-SPECIFIC AIP1B TRANSGENIC) MICE EXHIBITED AUGMENTED REACTIVE OXYGEN SPECIES PRODUCTION, EC ACTIVATION, AND NEOINTIMA FORMATION IN VASCULAR REMODELING MODELS. CONCLUSIONS: OUR CURRENT STUDY SUGGESTS THAT A SHIFT FROM ANTI-INFLAMMATORY AIP1A TO PROINFLAMMATORY AIP1B DURING CHRONIC INFLAMMATION PLAYS A KEY ROLE IN INFLAMMATORY VASCULAR DISEASES. 2020 20 5763 12 SOME COMMENTS ON MASOCHISM AND THE DELUSION OF OMNIPOTENCE FROM A DEVELOPMENTAL PERSPECTIVE. THIS PAPER EXPLORES THE RELATION OF THE DELUSION OF OMNIPOTENCE TO MASOCHISM AND SUGGESTS THAT THIS FANTASY CONSTITUTES A MAJOR COMPONENT OF THE RESISTANCE SO PROMINENT IN WORK WITH MASOCHISTIC PATIENTS. THE CONNECTIONS AMONG MASOCHISM, OMNIPOTENCE, NEGATIVE THERAPEUTIC REACTION, AND CLINGING TO PAIN ARE DISCUSSED. THE CLASSICAL VIEW HAS BEEN THAT THE FAILURE OF INFANTILE OMNIPOTENCE FORCES THE CHILD TO TURN TO REALITY. OUR EXPERIENCE WITH MASOCHISTIC PATIENTS SUGGESTS THAT IT IS THE REAL FAILURE TO ACHIEVE COMPETENT INTERACTIONS WITH OTHERS THAT FORCES THE CHILD TO TURN TO OMNIPOTENT SOLUTIONS. THE DISTINCTION IS MADE BETWEEN FANTASIES THAT ENHANCE THE REAL CAPACITIES OF THE SELF AND THOSE AIMED AT DENYING AND TRANSFORMING THE PAIN AND INADEQUACY OF THE MOTHER-CHILD RELATIONSHIP. THE EPIGENETIC TRANSFORMATIONS OF OMNIPOTENT FANTASIES THROUGH ALL LEVELS OF DEVELOPMENT ARE DESCRIBED. THE PATIENT'S NEED TO PROTECT THE OMNIPOTENT FANTASY IS DISCUSSED IN RELATION TO RESISTANCE AT EACH PHASE OF ANALYSIS. 1991