1 3336 142 HISTONE DEACETYLASE INHIBITORS INDUCE PROTEOLYSIS OF ACTIVATED CDC42-ASSOCIATED KINASE-1 IN LEUKEMIC CELLS. PURPOSE: ACTIVATED CDC42-ASSOCIATED KINASE-1 (ACK1/TNK2) AND EPIGENETIC REGULATORS OF THE HISTONE DEACETYLASE (HDAC) FAMILY REGULATE THE PROLIFERATION AND SURVIVAL OF LEUKEMIC CELLS. 18 HDACS FALL INTO FOUR CLASSES (I-IV). WE TESTED THE IMPACT OF CLINICALLY RELEVANT HISTONE DEACETYLASE INHIBITORS (HDACI) ON ACK1 AND IF SUCH DRUGS COMBINE FAVORABLY WITH THE THERAPEUTICALLY USED ACK1 INHIBITOR DASATINIB. METHODS: WE APPLIED THE BROAD-RANGE HDACI PANOBINOSTAT/LBH589 AND THE CLASS I HDAC-SPECIFIC INHIBITOR ENTINOSTAT/MS-275 TO VARIOUS ACUTE AND CHRONIC MYELOID LEUKEMIA CELLS (AML/CML). WE ALSO USED THE REPLICATIVE STRESS INDUCER HYDROXYUREA (HU), A STANDARD DRUG FOR LEUKEMIC PATIENTS, AND THE APOPTOSIS INDUCER STAUROSPORINE (STS). TO ASSESS CYTOTOXIC EFFECTS OF HDACI, WE MEASURED CELL CYCLE PROFILES AND DNA FRAGMENTATION BY FLOW CYTOMETRY. WESTERN BLOT WAS EMPLOYED TO ANALYZE PROTEIN EXPRESSION AND PHOSPHORYLATION. RESULTS: LBH589 AND MS-275 INDUCE PROTEOLYSIS OF ACK1 IN CML AND AML CELLS. PANOBINOSTAT MORE STRONGLY INDUCES APOPTOSIS THAN ENTINOSTAT, AND THIS CORRELATES WITH A SIGNIFICANTLY PRONOUNCED LOSS OF ACK1. STS AND HU ALSO PROPEL THE DEGRADATION OF ACK1 IN LEUKEMIC CELLS. MOREOVER, THE CASPASE INHIBITOR Z-VAD-FMK REDUCES ACK1 DEGRADATION IN THE PRESENCE OF HDACI. CONCOMITANT WITH THE ATTENUATION OF ACK1, WE NOTICED DECREASED PHOSPHORYLATION OF STAT3. DIRECT INHIBITION OF ACK1 WITH DASATINIB ALSO SUPPRESSES STAT3 PHOSPHORYLATION. FURTHERMORE, DASATINIB AND HDACI COMBINATIONS ARE EFFECTIVE AGAINST CML CELLS. CONCLUSION: HDACS SUSTAIN THE ACK1-STAT3 SIGNALING NODE AND LEUKEMIC CELL GROWTH. CONSISTENT WITH THEIR DIFFERENT EFFECTS ON ACK1 STABILITY OR AUTO-PHOSPHORYLATION, DASATINIB AND HDACI COMBINATIONS PRODUCE BENEFICIAL ANTILEUKEMIC EFFECTS. 2016 2 671 36 BOTHROPS MOOJENI L-AMINO ACID OXIDASE INDUCES APOPTOSIS AND EPIGENETIC MODULATION ON BCR-ABL(+) CELLS. BACKGROUND: RESISTANCE TO APOPTOSIS IN CHRONIC MYELOID LEUKEMIA (CML) IS ASSOCIATED WITH CONSTITUTIVE TYROSINE KINASE ACTIVITY OF THE BCR-ABL ONCOPROTEIN. THE DEREGULATED EXPRESSION OF APOPTOSIS-RELATED GENES AND ALTERATION IN EPIGENETIC MACHINERY MAY ALSO CONTRIBUTE TO APOPTOSIS RESISTANCE IN CML. TYROSINE KINASE INHIBITORS TARGET THE BCR-ABL ONCOPROTEIN AND ARE USED IN CML TREATMENT. THE RESISTANCE OF CML PATIENTS TO TYROSINE KINASE INHIBITORS HAS GUIDED THE SEARCH FOR NEW COMPOUNDS THAT MAY INDUCE APOPTOSIS IN BCR-ABL(+) LEUKEMIC CELLS AND IMPROVE THE DISEASE TREATMENT. METHODS: IN THE PRESENT STUDY, WE INVESTIGATED WHETHER THE L-AMINO ACID OXIDASE ISOLATED FROM BOTHROPS MOOJENI SNAKE VENOM (BMOOLAAO-I) (I) WAS CYTOTOXIC TO BCR-ABL(+) CELL LINES (HL-60.BCR-ABL, K562-S, AND K562-R), HL-60 (ACUTE PROMYELOCYTIC LEUKEMIA) CELLS, THE NON-TUMOR CELL LINE HEK-293, AND PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMC); AND (II) AFFECTED EPIGENETIC MECHANISMS, INCLUDING DNA METHYLATION AND MICRORNAS EXPRESSION IN VITRO. RESULTS: BMOOLAAO-I INDUCED ROS PRODUCTION, APOPTOSIS, AND DIFFERENTIAL DNA METHYLATION PATTERN OF REGULATORY APOPTOSIS GENES. THE TOXIN UPREGULATED EXPRESSION OF THE PRO-APOPTOTIC GENES BID AND FADD AND DOWNREGULATED DFFA EXPRESSION IN LEUKEMIC CELL LINES, AS WELL AS INCREASED MIR-16 EXPRESSION - WHOSE MAJOR PREDICTED TARGET IS THE ANTI-APOPTOTIC GENE BCL2 - IN BCR-ABL(+) CELLS. CONCLUSION: BMOOLAAO-I EXERTS SELECTIVE ANTITUMOR ACTION MEDIATED BY H(2)O(2) RELEASE AND INDUCES APOPTOSIS, AND ALTERATIONS IN EPIGENETIC MECHANISMS. THESE RESULTS SUPPORT FUTURE INVESTIGATIONS ON THE EFFECT OF BMOOLAAO-I ON IN VIVO MODELS TO DETERMINE ITS POTENTIAL IN CML THERAPY. 2020 3 5934 34 TARGETING FEATURES OF CURAXIN CBL0137 ON HEMATOLOGICAL MALIGNANCIES IN VITRO AND IN VIVO. THE ANTICANCER ACTIVITY OF CURAXIN CBL0137, A DNA-BINDING SMALL MOLECULE WITH CHROMATIN REMODULATING EFFECT, HAS BEEN DEMONSTRATED IN DIFFERENT CANCERS. HEREIN, A COMPARATIVE EVALUATION OF CBL0137 ACTIVITY WAS PERFORMED IN RESPECT TO ACUTE MYELOID LEUKEMIA (AML), ACUTE LYMPHOBLASTIC LEUKEMIA (ALL), CHRONIC MYELOID LEUKEMIA AND MULTIPLE MYELOMA (MM) CULTURED IN VITRO. MTT ASSAY SHOWED AML AND MM HIGHER SENSITIVITY TO CBL0137'S CYTOSTATIC EFFECT COMPARATIVELY TO OTHER HEMATOLOGICAL MALIGNANCY CELLS. FLOW CYTOMETRY CELL CYCLE ANALYSIS REVEALED AN INCREASE IN SUBG1 AND G2/M POPULATIONS AFTER CBL0137 CELL TREATMENT, BUT THE PREVALENT TYPE OF ARREST VARIED. APOPTOSIS ACTIVATION BY CBL0137 MEASURED BY ANNEXIN-V/PI DUAL STAINING WAS MORE ACTIVE IN AML AND MM CELLS. RT2 PCR ARRAY SHOWED THAT CHANGES CAUSED BY CBL0137 IN SIGNALING PATHWAYS INVOLVED IN CANCER PATHOGENESIS WERE MORE INTENSIVE IN AML AND MM CELLS. ON THE MURINE MODEL OF AML WEHI-3, CBL0137 SHOWED SIGNIFICANT ANTICANCER EFFECTS IN VIVO, WHICH WERE EVALUATED BY CORRESPONDING CHANGES IN SPLEEN AND LIVER. THUS, MORE PRONOUNCED ANTICANCER EFFECTS OF CBL0137 IN VITRO WERE OBSERVED IN RESPECT TO AML AND MM. EXPERIMENTS IN VIVO ALSO INDICATED THE PERSPECTIVE OF CBL0137 USE FOR AML TREATMENT. THIS IN ACCORDANCE WITH THE FRONTLINE TREATMENT APPROACH IN AML USING EPIGENETIC DRUGS. 2023 4 5775 31 SPERMIDINE/SPERMINE N(1)-ACETYLTRANSFERASE ACTIVITY ASSOCIATES WITH WHITE BLOOD CELL COUNT IN MYELOID LEUKEMIAS. THE METABOLISM OF POLYAMINES, THE CATIONIC SMALL MOLECULES ESSENTIAL FOR CELL PROLIFERATION AND DIFFERENTIATION, IS ALTERED IN CANCER CELLS AND CAN BE EXPLOITED IN CANCER DIAGNOSIS AND THERAPY. SPERMIDINE/SPERMINE N(1)-ACETYLTRANSFERASE (SSAT), WHICH REGULATES INTRACELLULAR LEVELS OF POLYAMINES BY CATABOLIZING SPERMIDINE AND SPERMINE, HAS A CONTROVERSIAL ROLE IN THE DEVELOPMENT OF CANCERS. IN THIS STUDY, THE POLYAMINE METABOLISM AND FUNCTION OF SSAT WERE CHARACTERIZED IN ACUTE MYELOID LEUKEMIA (AML), CHRONIC MYELOID LEUKEMIA (CML), AND ACUTE LYMPHOID LEUKEMIA PATIENT SAMPLES. ALSO, MICE OVEREXPRESSING SSAT AND HAVING A MYELOPROLIFERATIVE PHENOTYPE WERE ANALYZED FOR THEIR RESPONSE TO DECITABINE AND HISTONE DEACETYLASE INHIBITOR TRICHOSTATIN A. THE PRESENCE OF EPIGENETIC FACTORS IN THE BONE MARROW CELLS OF SSAT MICE WAS ANALYZED. ELEVATED LEVELS OF SPERMIDINE AND SPERMINE, AS WELL AS INCREASED ACTIVITY OF SSAT, WERE DETECTED IN AML, CML, AND ACUTE LYMPHOID LEUKEMIA PATIENTS COMPARED WITH THE CONTROLS. HOWEVER, WE FOUND SSAT ACTIVITY TO BE ASSOCIATED WITH WHITE BLOOD CELL COUNT ONLY IN AML AND CML PATIENTS. DECITABINE TREATMENT BROUGHT THE PERIPHERAL BLOOD AND BONE MARROW CELL COUNTS OF SSAT MICE TO THE LEVEL OF WILD-TYPE MICE. SPERMIDINE/SPERMINE N(1)-ACETYLTRANSFERASE MICE HAD INCREASED HISTONE METHYLATION AND AN INCREASED LEVEL OF HISTONE DEACETYLASE 1 IN THEIR BONE MARROW CELLS. THE STUDY SUGGESTS THAT SSAT INFLUENCES THE DEVELOPMENT OF MYELOID MALIGNANCIES, AND EPIGENETIC FACTORS PARTLY CONTRIBUTE TO THE SSAT OVEREXPRESSION-INDUCED MYELOPROLIFERATIVE DISEASE IN MICE. 2014 5 1843 29 EFFECTS OF TELOMERASE INHIBITOR ON EPIGENETIC CHROMATIN MODIFICATION ENZYMES IN MALIGNANCIES. TELOMERASE HAS A CRITICAL ROLE IN CELL PROLIFERATION, TUMOR MAINTAINING, AND THERAPY RESISTANCE, WHICH ACT BY MODIFYING MANY SIGNALING PATHWAYS. 2-[(E)-3-NAPHTALEN-2-YL-BUT-2-ENOYLAMINO]-BENZOIC ACID (BIBR1532) IS ONE OF THE MOST STUDIED TELOMERASE INHIBITORS, AND IT TARGETS TELOMERASE COMPONENTS TERC AND TERT. IN THIS NOVEL STUDY, WE AIMED TO INVESTIGATE THE EPIGENETIC EFFECTS OF BIBR1532 ON BOTH HEMATOLOGIC MALIGNANCIES AND SOLID TUMORS. K-562 HUMAN CHRONIC MYELOID LEUKEMIA CELL LINE AND U87MG GLIOBLASTOMA CELL LINE WERE COMPARED WITH CONTROL GROUPS WITHOUT BIBR1532 TREATMENT. CYTOTOXIC EFFECTS OF BIBR1532 WERE DETERMINED BY USING WST-1 ASSAY. APOPTOTIC EFFECTS OF BIBR1532 WERE DETECTED BY USING ANNEXIN V METHOD. TO ASSESS EXPRESSION CHANGES IN THE HUMAN EPIGENETIC CHROMATIN MODIFICATION ENZYME GENES, TOTAL RNA WAS ISOLATED FROM K-562 AND U87MG CELLS TREATED WITH BIBR1532 AND UNTREATED CONTROL CELLS. BIBR1532 INDUCED 2.41-FOLD APOPTOTIC CELL DEATH IN U87MG CELL LINES COMPARED WITH CONTROL GROUPS. APOPTOSIS WAS SLIGHTLY INDUCED IN K-562 CELLS WITH BIBR1532 TREATMENT COMPARED WITH CONTROL CELLS. WE OBSERVED THAT BIBR1532 ALSO REGULATES SIMILAR GENES IN BOTH CELL LINES, AND IT IS USEFUL ON EPIGENETIC MECHANISMS. AS A RESULT, TELOMERASE INHIBITOR BIBR1532 HAS A SIGNIFICANT EFFECT ON BOTH HEMATOLOGICAL MALIGNANCIES AND SOLID TUMORS. 2018 6 2763 27 EXPRESSION OF THE LEUKEMIC PROGNOSTIC MARKER CD7 IS LINKED TO EPIGENETIC MODIFICATIONS IN CHRONIC MYELOID LEUKEMIA. BACKGROUND: EXPRESSION LEVELS OF THE CELL SURFACE GLYCOPROTEIN, CD7, AND THE SERINE PROTEASE, ELASTASE 2 (ELA2), IN THE LEUKEMIC CELLS OF PATIENTS WITH CHRONIC MYELOID LEUKEMIA (CML) HAVE BEEN ASSOCIATED WITH CLINICAL OUTCOME. HOWEVER, LITTLE IS KNOWN ABOUT THE MECHANISMS THAT UNDERLIE THE VARIABLE EXPRESSION OF THESE GENES IN THE LEUKEMIC CELLS. RESULTS: TO ADDRESS THIS QUESTION, WE COMPARED THE LEVEL OF THEIR EXPRESSION WITH THE DNA METHYLATION AND HISTONE ACETYLATION STATUS OF 5' SEQUENCES OF BOTH GENES IN LEUKEMIC CELL LINES AND PRIMITIVE (LIN-CD34+) LEUKEMIC CELLS FROM CHRONIC PHASE CML PATIENTS. DNA METHYLATION OF THE ELA2 GENE PROMOTER DID NOT CORRELATE WITH ITS EXPRESSION PATTERN IN LIN-CD34+ CELLS FROM CHRONIC PHASE CML PATIENT SAMPLES EVEN THOUGH THERE WAS CLEAR DIFFERENTIAL DNA METHYLATION OF THIS LOCUS IN ELA2-EXPRESSING AND NON-EXPRESSING CELL LINES. IN CONTRAST, WE FOUND A STRONG RELATION BETWEEN CD7 EXPRESSION AND TRANSCRIPTION-PERMISSIVE CHROMATIN MODIFICATIONS, BOTH AT THE LEVEL OF DNA METHYLATION AND HISTONE ACETYLATION WITH EVIDENCE OF HYPOMETHYLATION OF THE CD7 PROMOTER REGION IN THE LIN-CD34+ CELLS FROM CML PATIENTS WITH HIGH CD7 EXPRESSION. CONCLUSION: THESE FINDINGS INDICATE A LINK BETWEEN EPIGENETIC MODIFICATIONS AND CD7 EXPRESSION IN PRIMITIVE CML CELLS. 2010 7 3484 36 IDENTIFICATION OF CHROMATIN REMODELING GENES ARID4A AND ARID4B AS LEUKEMIA SUPPRESSOR GENES. BACKGROUND: LEUKEMIA EVOLVES THROUGH A MULTISTEP PROCESS FROM PREMALIGNANCY TO MALIGNANCY. EPIGENETIC ALTERATIONS, INCLUDING HISTONE MODIFICATIONS, HAVE BEEN PROPOSED TO PLAY AN IMPORTANT ROLE IN TUMORIGENESIS. THE INVOLVEMENT OF TWO CHROMATIN REMODELING GENES, RETINOBLASTOMA-BINDING PROTEIN 1 (RBBP1/ARID4A) AND RBBP1-LIKE 1 (RBBP1L1/ARID4B), IN LEUKEMOGENESIS WAS NOT CHARACTERIZED. METHODS: THE LEUKEMIC PHENOTYPE OF MICE DEFICIENT FOR ARID4A WITH OR WITHOUT HAPLOINSUFFICIENCY FOR ARID4B WAS INVESTIGATED BY SERIALLY MONITORING COMPLETE BLOOD COUNTS TOGETHER WITH MICROSCOPIC HISTOLOGIC ANALYSIS AND FLOW CYTOMETRIC ANALYSIS OF BONE MARROW AND SPLEEN FROM THE ARID4A(-/-) MICE OR ARID4A(-/-)ARID4B(+/-) MICE. REGULATION IN BONE MARROW CELLS OF DOWNSTREAM GENES IMPORTANT FOR NORMAL HEMATOPOIESIS WAS ANALYZED BY REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION. GENOTYPIC EFFECTS ON HISTONE MODIFICATIONS WERE EXAMINED BY WESTERN BLOTTING AND IMMUNOFLUORESCENCE ANALYSIS. ALL STATISTICAL TESTS WERE TWO-SIDED. RESULTS: YOUNG (2-5 MONTHS OLD) ARID4A-DEFICIENT MICE HAD INEFFECTIVE BLOOD CELL PRODUCTION IN ALL HEMATOPOIETIC LINEAGES. BEYOND 5 MONTHS OF AGE, THE ARID4A(-/-) MICE MANIFESTED MONOCYTOSIS, ACCOMPANIED BY SEVERE ANEMIA AND THROMBOCYTOPENIA. THESE SICK ARID4A(-/-) MICE SHOWED BONE MARROW FAILURE WITH MYELOFIBROSIS ASSOCIATED WITH SPLENOMEGALY AND HEPATOMEGALY. FIVE OF 42 ARID4A(-/-) MICE AND 10 OF 12 ARID4A(-/-)ARID4B(+/-) MICE PROGRESSED TO ACUTE MYELOID LEUKEMIA (AML) AND HAD RAPID FURTHER INCREASES OF LEUKOCYTE COUNTS. EXPRESSION OF HOX GENES (HOXB3, HOXB5, HOXB6, AND HOXB8) WAS DECREASED IN ARID4A-DEFICIENT BONE MARROW CELLS WITH OR WITHOUT ARID4B HAPLOINSUFFICIENCY, AND FOXP3 EXPRESSION WAS REDUCED IN ARID4A(-/-)ARID4B(+/-) BONE MARROW. INCREASES OF HISTONE TRIMETHYLATION OF H3K4, H3K9, AND H4K20 (FOLD INCREASES IN TRIMETHYLATION = 32, 95% CONFIDENCE INTERVAL [CI] = 27 TO 32; 45, 95% CI = 41 TO 49; AND 2.2, 95% CI = 1.7 TO 2.7, RESPECTIVELY) WERE OBSERVED IN THE BONE MARROW OF ARID4A-DEFICIENT MICE. CONCLUSIONS: ARID4A-DEFICIENT MICE INITIALLY DISPLAY INEFFECTIVE HEMATOPOIESIS, FOLLOWED BY TRANSITION TO CHRONIC MYELOMONOCYTIC LEUKEMIA (CMML)-LIKE MYELODYSPLASTIC/MYELOPROLIFERATIVE DISORDER, AND THEN TRANSFORMATION TO AML. THE DISEASE PROCESSES IN THE ARID4A-DEFICIENT MICE ARE VERY SIMILAR TO THE COURSE OF EVENTS IN HUMANS WITH CMML AND AML. THIS MOUSE MODEL HAS THE POTENTIAL TO FURNISH ADDITIONAL INSIGHTS INTO THE ROLE OF EPIGENETIC ALTERATIONS IN LEUKEMOGENESIS, AND IT MAY BE USEFUL IN DEVELOPING NOVEL PHARMACOLOGICAL APPROACHES TO TREATMENT OF PRELEUKEMIC AND LEUKEMIC STATES. 2008 8 3128 39 GIPC-REGULATED IGFBP-3 PROMOTES HSC MIGRATION IN VITRO AND PORTAL HYPERTENSION IN VIVO THROUGH A BETA1-INTEGRIN PATHWAY. BACKGROUND & AIMS: TRANSFORMING GROWTH FACTOR (TGF-BETA)-INDUCED ACTIVATION OF QUIESCENT HEPATIC STELLATE CELLS (HSCS) AND THEIR TRANSFORMATION TO MYOFIBROBLASTS IS A KEY EVENT IN LIVER FIBROSIS AND PORTAL HYPERTENSION. GIPC (ALSO REFERRED TO AS SYNECTIN) IS A DOWNSTREAM SIGNAL ACTIVATION MOLECULE OF TGF-BETA AND OTHER RECEPTORS. IN THIS STUDY, WE SOUGHT TO IDENTIFY NOVEL GENES TARGETED BY TGF-BETA AND GIPC AND ELUCIDATE IF AND HOW THEY MAY CONTRIBUTE TO LIVER FIBROSIS. METHODS: WE PERFORMED SEQUENTIAL MESSENGER RNA SEQUENCING ANALYSIS ON TGF-BETA-STIMULATED HSCS AND THEN ON TGF-BETA-STIMULATED HSCS IN THE PRESENCE AND ABSENCE OF GIPC ALSO REFERRED TO AS SYNECTIN (GIPC) KNOCKDOWN. INSULIN-LIKE GROWTH FACTOR BINDING PROTEIN-3 (IGFBP-3) TRANSPORT PROTEIN EMERGED AS A TOP ACTIVATION TARGET OF BOTH TGF-BETA AND GIPC. QUANTITATIVE POLYMERASE CHAIN REACTION, ENZYME-LINKED IMMUNOSORBENT ASSAY, TARGETED CHROMATIN IMMUNOPRECIPITATION, AND WESTERN BLOT ANALYSIS WERE DONE FOR FURTHER CONFIRMATION. RESULTS: IGFBP-3, AN INSULIN GROWTH FACTOR TRANSPORT PROTEIN, EMERGED AS A TOP ACTIVATION TARGET OF BOTH TGF-BETA AND GIPC, WHICH WAS CONFIRMED BY QUANTITATIVE POLYMERASE CHAIN REACTION, ENZYME-LINKED IMMUNOSORBENT ASSAY, AND WESTERN BLOT ANALYSIS. TARGETED CHROMATIN IMMUNOPRECIPITATION SHOWED THAT GIPC INCREASES THE HISTONE 3 LYSINE 27 (H3K27) ACETYLATION ACTIVATING MARK AND CONCURRENTLY DECREASES THE H3K27 INHIBITORY TRIMETHYLATION (H3K27M3) MARK, PROVIDING AN EPIGENETIC CORRELATE TO THE GENE REGULATION CHANGES. IN VIVO, GLOBAL KNOCKOUT OF IGFBP-3 MICE RESULTED IN ATTENUATION OF HSC ACTIVATION MARKERS AND ATTENUATION OF PORTAL PRESSURE IN RESPONSE TO CHRONIC LIVER INJURY MODELS. ANALYSIS OF SERUM LEVELS FROM CIRRHOTIC PATIENTS ALSO SHOWED AN IGFBP-3 INCREASE OF MORE THAN 2-FOLD COMPARED WITH HEALTHY CONTROLS. FINALLY, IN VITRO MECHANISM STUDIES SHOWED THAT IGFBP-3 PROMOTES HSC MIGRATION THROUGH INTEGRIN-DEPENDENT PHOSPHORYLATION OF PROTEIN KINASE B. CONCLUSIONS: TGF-BETA UP-REGULATES IGFBP-3 THROUGH GIPC, LEADING TO INCREASED HSC MIGRATION IN VITRO AND PROMOTES PORTAL HYPERTENSION IN VIVO. THESE STUDIES SUPPORT THE ROLE OF IGFBP-3 AS A POTENTIAL PATHOPHYSIOLOGIC TARGET OR BIOMARKER IN CHRONIC LIVER DISEASE. 2020 9 1334 31 DEREGULATION OF AIOLOS EXPRESSION IN CHRONIC LYMPHOCYTIC LEUKEMIA IS ASSOCIATED WITH EPIGENETIC MODIFICATIONS. CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS CHARACTERIZED BY A CLONAL ACCUMULATION OF MATURE NEOPLASTIC B CELLS THAT ARE RESISTANT TO APOPTOSIS. AIOLOS, A MEMBER OF THE IKAROS FAMILY OF ZINC-FINGER TRANSCRIPTION FACTORS, PLAYS AN IMPORTANT ROLE IN THE CONTROL OF MATURE B LYMPHOCYTE DIFFERENTIATION AND MATURATION. IN THIS STUDY, WE SHOWED THAT AIOLOS EXPRESSION IS UP-REGULATED IN B-CLL CELLS. THIS OVEREXPRESSION DOES NOT IMPLICATE ISOFORM IMBALANCE OR DISTURB AIOLOS SUBCELLULAR LOCALIZATION. THE CHROMATIN STATUS AT THE AIOLOS PROMOTER IN CLL IS DEFINED BY THE DEMETHYLATION OF DNA AND AN ENRICHMENT OF EUCHROMATIN ASSOCIATED HISTONE MARKERS, SUCH AS THE DIMETHYLATION OF THE LYSINE 4 ON HISTONE H3. THESE EPIGENETIC MODIFICATIONS SHOULD ALLOW ITS UPSTREAM EFFECTORS, SUCH AS NUCLEAR FACTOR-KAPPAB, CONSTITUTIVELY ACTIVATED IN CLL, TO GAIN ACCESS TO PROMOTER, RESULTING UP-REGULATION OF AIOLOS. TO DETERMINE THE CONSEQUENCES OF AIOLOS DEREGULATION IN CLL, WE ANALYZED THE EFFECTS OF AIOLOS OVEREXPRESSION OR DOWN-REGULATION ON APOPTOSIS. AIOLOS IS INVOLVED IN CELL SURVIVAL BY REGULATING THE EXPRESSION OF SOME BCL-2 FAMILY MEMBERS. OUR RESULTS STRONGLY SUGGEST THAT AIOLOS DEREGULATION BY EPIGENETIC MODIFICATIONS MAY BE A HALLMARK OF CLL. 2011 10 197 26 ACINAR ATP8B1/LPC PATHWAY PROMOTES MACROPHAGE EFFEROCYTOSIS AND CLEARANCE OF INFLAMMATION DURING CHRONIC PANCREATITIS DEVELOPMENT. NONINFLAMMATORY CLEARANCE OF DYING CELLS BY PROFESSIONAL PHAGOCYTES, TERMED EFFEROCYTOSIS, IS FUNDAMENTAL IN BOTH HOMEOSTASIS AND INFLAMMATORY FIBROSIS DISEASE BUT HAS NOT BEEN CONFIRMED TO OCCUR IN CHRONIC PANCREATITIS (CP). HERE, WE INVESTIGATED WHETHER EFFEROCYTOSIS CONSTITUTES A NOVEL REGULATORY TARGET IN CP AND ITS MECHANISMS. PRSS1 TRANSGENIC (PRSS1(TG)) MICE WERE TREATED WITH CAERULEIN TO MIMIC CP DEVELOPMENT. PHOSPHOLIPID METABOLITE PROFILING AND EPIGENETIC ASSAYS WERE PERFORMED WITH PRSS1(TG) CP MODELS. THE POTENTIAL FUNCTIONS OF ATP8B1 IN CP MODEL WERE CLARIFIED USING ATP8B1-OVEREXPRESSING ADENO-ASSOCIATED VIRUS, IMMUNOFLUORESCENCE, ENZYME-LINKED IMMUNOSORBENT ASSAY(ELISA), AND LIPID METABOLOMIC APPROACHES. ATAC-SEQ COMBINED WITH RNA-SEQ WAS THEN USED TO IDENTIFY TRANSCRIPTION FACTORS BINDING TO THE ATP8B1 PROMOTER, AND CHIP-QPCR AND LUCIFERASE ASSAYS WERE USED TO CONFIRM THAT THE IDENTIFIED TRANSCRIPTION FACTOR BOUND TO THE ATP8B1 PROMOTER, AND TO IDENTIFY THE SPECIFIC BINDING SITE. FLOW CYTOMETRY WAS PERFORMED TO ANALYZE THE PROPORTION OF PANCREATIC MACROPHAGES. DECREASED EFFEROCYTOSIS WITH AGGRAVATED INFLAMMATION WAS IDENTIFIED IN CP. THE LYSOPHOSPHATIDYLCHOLINE (LPC) PATHWAY WAS THE MOST OBVIOUSLY DYSREGULATED PHOSPHOLIPID PATHWAY, AND LPC AND ATP8B1 EXPRESSION GRADUALLY DECREASED DURING CP DEVELOPMENT. H3K27ME3 CHIP-SEQ SHOWED THAT INCREASED ATP8B1 PROMOTER METHYLATION LED TO TRANSCRIPTIONAL INHIBITION. ATP8B1 COMPLEMENTATION SUBSTANTIALLY INCREASED THE LPC CONCENTRATION AND IMPROVED CP OUTCOMES. BHLHA15 WAS IDENTIFIED AS A TRANSCRIPTION FACTOR THAT BINDS TO THE ATP8B1 PROMOTER AND REGULATES PHOSPHOLIPID METABOLISM. OUR STUDY INDICATES THAT THE ACINAR ATP8B1/LPC PATHWAY ACTS AS AN IMPORTANT "FIND-ME" SIGNAL FOR MACROPHAGES AND PLAYS A PROTECTIVE ROLE IN CP, WITH ATP8B1 TRANSCRIPTION PROMOTED BY THE ACINAR CELL-SPECIFIC TRANSCRIPTION FACTOR BHLHA15. BHLHA15, ATP8B1, AND LPC COULD BE CLINICALLY TRANSLATED INTO VALUABLE THERAPEUTIC TARGETS TO OVERCOME THE LIMITATIONS OF CURRENT CP THERAPIES. 2022 11 4696 27 NF-KAPPAB REPRESSES RETINOIC ACID RECEPTOR-MEDIATED GPRC5A TRANSACTIVATION IN LUNG EPITHELIAL CELLS TO PROMOTE NEOPLASIA. CHRONIC INFLAMMATION IS ASSOCIATED WITH LUNG TUMORIGENESIS, IN WHICH NF-KAPPAB-MEDIATED EPIGENETIC REGULATION PLAYS A CRITICAL ROLE. LUNG TUMOR SUPPRESSOR G PROTEIN-COUPLED RECEPTOR, FAMILY C, MEMBER 5A (GPRC5A), IS REPRESSED IN MOST NON-SMALL CELL LUNG CANCER (NSCLC); HOWEVER, THE MECHANISMS REMAIN UNCLEAR. HERE, WE SHOW THAT NF-KAPPAB ACTS AS A TRANSCRIPTIONAL REPRESSOR IN SUPPRESSION OF GPRC5A. NF-KAPPAB INDUCED GPRC5A REPRESSION BOTH IN VITRO AND IN VIVO. INTRIGUINGLY, TRANSACTIVATION OF NF-KAPPAB DOWNSTREAM TARGETS WAS NOT REQUIRED, BUT THE TRANSACTIVATION DOMAIN OF RELA/P65 WAS REQUIRED FOR GPRC5A REPRESSION. NF-KAPPAB DID NOT BIND TO ANY POTENTIAL CIS-ELEMENT IN THE GPRC5A PROMOTER. INSTEAD, P65 WAS COMPLEXED WITH RETINOIC ACID RECEPTOR ALPHA/BETA (RARALPHA/BETA) AND RECRUITED TO THE RA RESPONSE ELEMENT SITE AT THE GPRC5A PROMOTER, RESULTING IN DISRUPTED RNA POLYMERASE II COMPLEXING AND SUPPRESSED TRANSCRIPTION. NOTABLY, PHOSPHORYLATION ON SERINE 276 OF P65 WAS REQUIRED FOR INTERACTION WITH RARALPHA/BETA AND REPRESSION OF GPRC5A. MOREOVER, NF-KAPPAB-MEDIATED EPIGENETIC REPRESSION WAS THROUGH SUPPRESSION OF ACETYLATED HISTONE H3K9 (H3K9AC), BUT NOT DNA METHYLATION OF THE CPG ISLANDS, AT THE GPRC5A PROMOTER. CONSISTENTLY, A HISTONE DEACETYLASE INHIBITOR, BUT NOT DNA METHYLATION INHIBITOR, RESTORED GPRC5A EXPRESSION IN NSCLC CELLS. THUS, NF-KAPPAB INDUCES TRANSCRIPTIONAL REPRESSION OF GPRC5A VIA A COMPLEX WITH RARALPHA/BETA AND MEDIATES EPIGENETIC REPRESSION VIA SUPPRESSION OF H3K9AC. 2023 12 5101 28 POLYCOMB FACTOR PHF19 CONTROLS CELL GROWTH AND DIFFERENTIATION TOWARD ERYTHROID PATHWAY IN CHRONIC MYELOID LEUKEMIA CELLS. POLYCOMB GROUP (PCG) OF PROTEINS ARE A GROUP OF HIGHLY CONSERVED EPIGENETIC REGULATORS INVOLVED IN MANY BIOLOGICAL FUNCTIONS, SUCH AS EMBRYONIC DEVELOPMENT, CELL PROLIFERATION, AND ADULT STEM CELL DETERMINATION. PHD FINGER PROTEIN 19 (PHF19) IS AN ASSOCIATED FACTOR OF POLYCOMB REPRESSOR COMPLEX 2 (PRC2), OFTEN UPREGULATED IN HUMAN CANCERS. IN PARTICULAR, MYELOID LEUKEMIA CELL LINES SHOW INCREASED LEVELS OF PHF19, YET LITTLE IS KNOWN ABOUT ITS FUNCTION. HERE, WE HAVE CHARACTERIZED THE ROLE OF PHF19 IN MYELOID LEUKEMIA CELLS. WE DEMONSTRATED THAT PHF19 DEPLETION DECREASES CELL PROLIFERATION AND PROMOTES CHRONIC MYELOID LEUKEMIA (CML) DIFFERENTIATION. MECHANISTICALLY, WE HAVE SHOWN HOW PHF19 REGULATES THE PROLIFERATION OF CML THROUGH A DIRECT REGULATION OF THE CELL CYCLE INHIBITOR P21. FURTHERMORE, WE OBSERVED THAT MTF2, A PHF19 HOMOLOG, PARTIALLY COMPENSATES FOR PHF19 DEPLETION IN A SUBSET OF TARGET GENES, INSTRUCTING SPECIFIC ERYTHROID DIFFERENTIATION. TAKEN TOGETHER, OUR RESULTS SHOW THAT PHF19 IS A KEY TRANSCRIPTIONAL REGULATOR FOR CELL FATE DETERMINATION AND COULD BE A POTENTIAL THERAPEUTIC TARGET FOR MYELOID LEUKEMIA TREATMENT. 2021 13 1234 30 CRYPTOTANSHINONE SUPPRESSES KEY ONCO-PROLIFERATIVE AND DRUG-RESISTANT PATHWAYS OF CHRONIC MYELOID LEUKEMIA BY TARGETING STAT5 AND STAT3 PHOSPHORYLATION. C-MYC AND SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION (STAT) FAMILY PROTEINS HAVE BEEN PROPOSED TO BE IMPORTANT DOWNSTREAM GENES OF BCR-ABL, WHICH CHARACTERIZES MOST CASES OF CHRONIC MYELOID LEUKEMIA (CML). HERE, WE REPORT A C-MYC PATHWAY-TARGETED SCREENING OF SEVEN NATURAL ANTICANCER COMPOUNDS, IN WHICH WE IDENTIFIED CRYPTOTANSHINONE AS A HIGHLY PROMISING AGENT FOR CML THERAPY. CRYPTOTANSHINONE DEPLETES C-MYC IN CML BY REPRESSING THE PHOSPHORYLATION OF STAT5. DECREASED VIABILITY OF K562 CELLS CORRELATED WITH P-STAT5 SUPPRESSION. UNEXPECTEDLY, IMATINIB ACTIVATES RATHER THAN INHIBITS THE PHOSPHORYLATION OF STAT3 IN K562 CELLS. WE DEMONSTRATED THAT CRYPTOTANSHINONE, AS A DUAL INHIBITOR OF P-STAT5 AND P-STAT3, CAN EFFECTIVELY BLOCK IL-6-MEDIATED STAT3 ACTIVATION AND REVERSE BCR-ABL KINASE-INDEPENDENT DRUG RESISTANCE. MOREOVER, WE SHOWED THAT THE EPIGENETIC REBALANCE BETWEEN DECREASED BCR-ABL/STAT5/C-MYC AND ENHANCED STAT3/MULTI-DRUG RESISTANCE (MDR) PATHWAYS IS CHARACTERISTIC OF THE CANCER STEM CELL-LIKE PROPERTY OF K562/ADR. SIMULTANEOUSLY SUPPRESSING THESE TWO PATHWAYS USING CRYPTOTANSHINONE PROVES TO BE CRITICAL FOR THE MALIGNANT NETWORK REDRESS AND MDR REVERSAL OF K562/ADR. THESE STUDIES REVEAL THE DUAL FUNCTIONS OF CRYPTOTANSHINONE THAT SUPPRESS KEY ONCOGENIC PROLIFERATION AND DRUG-RESISTANT PATHWAYS IN CML CELLS BY TARGETING P-STAT5 AND P-STAT3, PROVIDING A NEW STRATEGY FOR CML THERAPY THAT TAKES ADVANTAGE OF NATURAL PRODUCTS. 2018 14 1951 37 EPIGENETIC ACTIVATION OF THE TUSC3 GENE AS A POTENTIAL THERAPY FOR XMEN DISEASE. BACKGROUND: X-LINKED MAGT1 DEFICIENCY WITH INCREASED SUSCEPTIBILITY TO EPSTEIN-BARR VIRUS INFECTION AND N-LINKED GLYCOSYLATION DEFECT (XMEN) DISEASE IS A RARE COMBINED IMMUNODEFICIENCY CAUSED BY LOSS-OF-FUNCTION MUTATIONS IN THE MAGNESIUM TRANSPORTER 1 (MAGT1) GENE. MAGT1 DEFICIENCY IMPAIRS MAGNESIUM TRANSPORT AND THE N-LINKED GLYCOSYLATION OF A PANEL OF PROTEINS, WHICH SUBSEQUENTLY ABOLISHES THE EXPRESSION OF KEY IMMUNE RECEPTORS SUCH AS NATURAL KILLER GROUP 2, MEMBER D (AKA NKG2D). THESE EFFECTS INDUCE IMMUNE SYSTEM ABNORMALITIES, CHRONIC EPSTEIN-BARR VIRUS INFECTION, AND NEOPLASIA. RECENT RESEARCH SHOWS THAT MAGT1 AND TUMOR CANDIDATE SUPPRESSOR 3 (TUSC3) SHARE HIGH SEQUENCE AND FUNCTIONAL SIMILARITY. OBJECTIVE: WE SOUGHT TO INVESTIGATE THE FEASIBILITY OF ACTIVATING TUSC3 EXPRESSION TO PROVIDE A POTENTIAL THERAPEUTIC STRATEGY FOR XMEN DISEASE. METHODS: THE EXPRESSION PROFILES OF MAGT1 AND TUSC3 WERE ANALYZED USING MULTIPLE DATABASES, REAL-TIME QUANTITATIVE PCR, AND WESTERN BLOT. THE EFFECTS OF DECITABINE AND PANOBINOSTAT ON THE REGULATION OF TUSC3 EXPRESSION WERE EXPLORED IN BOTH MAGT1 KNOCKOUT (KO)/PATIENT-DERIVED LYMPHOCYTES AND MAGT1 KO HEPATOCYTES. RESULTS: ALTHOUGH TUSC3 IS WIDELY EXPRESSED, IT IS UNDETECTABLE SPECIFICALLY IN THE IMMUNE SYSTEM AND LIVER, CONSISTENT WITH THE MAIN DISEASED TISSUES IN PATIENTS WITH XMEN DISEASE. CRISPR/CAS9-MEDIATED KO OF MAGT1 IN THE NKL CELL LINE SUCCESSFULLY MIMICKED THE PHENOTYPES OF XMEN PATIENT-DERIVED LYMPHOCYTES, AND EXOGENOUS EXPRESSION OF TUSC3 RESCUED THE DEFICIENCIES IN KO NKL CELLS. USING THIS IN VITRO MODEL, WE IDENTIFIED 2 EPIGENETIC DRUGS, DECITABINE AND PANOBINOSTAT, BY SCREENING. COMBINATION TREATMENT USING THESE 2 DRUGS SIGNIFICANTLY UPREGULATED TUSC3 EXPRESSION AND RESCUED THE IMMUNE AND LIVER ABNORMALITIES. CONCLUSIONS: EPIGENETIC ACTIVATION OF TUSC3 EXPRESSION CONSTITUTES AN EFFECTIVE THERAPEUTIC STRATEGY FOR XMEN DISEASE. 2023 15 6659 24 UPREGULATION OF AKT3 CONFERS RESISTANCE TO THE AKT INHIBITOR MK2206 IN BREAST CANCER. ACQUIRED RESISTANCE TO MOLECULAR TARGETED THERAPY REPRESENTS A MAJOR CHALLENGE FOR THE EFFECTIVE TREATMENT OF CANCER. HYPERACTIVATION OF THE PI3K/AKT PATHWAY IS FREQUENTLY OBSERVED IN VIRTUALLY ALL HUMAN MALIGNANCIES, AND NUMEROUS PI3K AND AKT INHIBITORS ARE CURRENTLY UNDER CLINICAL EVALUATION. HOWEVER, MECHANISMS OF ACQUIRED RESISTANCE TO AKT INHIBITORS HAVE YET TO BE DESCRIBED. HERE, WE USE A BREAST CANCER PRECLINICAL MODEL TO IDENTIFY RESISTANCE MECHANISMS TO A SMALL MOLECULE ALLOSTERIC AKT INHIBITOR, MK2206. USING A STEP-WISE AND CHRONIC HIGH-DOSE EXPOSURE, BREAST CANCER CELL LINES HARBORING ONCOGENIC PI3K RESISTANT TO MK2206 WERE ESTABLISHED. USING THIS MODEL, WE REVEAL THAT AKT3 EXPRESSION IS MARKEDLY UPREGULATED IN AKT INHIBITOR-RESISTANT CELLS. INDUCTION OF AKT3 IS REGULATED EPIGENETICALLY BY THE BROMODOMAIN AND EXTRA TERMINAL DOMAIN PROTEINS. IMPORTANTLY, KNOCKDOWN OF AKT3, BUT NOT AKT1 OR AKT2, IN RESISTANT CELLS RESTORES SENSITIVITY TO MK2206. AKT INHIBITOR-RESISTANT CELLS ALSO DISPLAY AN EPITHELIAL TO MESENCHYMAL TRANSITION PHENOTYPE AS ASSESSED BY ALTERATIONS IN THE LEVELS OF E-CADHERIN, N-CADHERIN, AND VIMENTIN, AS WELL AS ENHANCED INVASIVENESS OF TUMOR SPHEROIDS. NOTABLY, THE INVASIVE MORPHOLOGY OF RESISTANT SPHEROIDS IS DIMINISHED UPON AKT3 DEPLETION. WE ALSO SHOW THAT RESISTANCE TO MK2206 IS REVERSIBLE BECAUSE UPON DRUG REMOVAL RESISTANT CELLS REGAIN SENSITIVITY TO AKT INHIBITION, ACCOMPANIED BY REEXPRESSION OF EPITHELIAL MARKERS AND REDUCTION OF AKT3 EXPRESSION, IMPLYING THAT EPIGENETIC REPROGRAMMING CONTRIBUTES TO ACQUISITION OF RESISTANCE. THESE FINDINGS PROVIDE A RATIONALE FOR DEVELOPING THERAPEUTICS TARGETING AKT3 TO CIRCUMVENT ACQUIRED RESISTANCE IN BREAST CANCER. MOL CANCER THER; 15(8); 1964-74. (C)2016 AACR. 2016 16 368 27 AMYLOID BETA-MEDIATED EPIGENETIC ALTERATION OF INSULIN-LIKE GROWTH FACTOR BINDING PROTEIN 3 CONTROLS CELL SURVIVAL IN ALZHEIMER'S DISEASE. SWEDISH DOUBLE MUTATION (KM670/671NL) OF AMYLOID PRECURSOR PROTEIN (APP) IS REPORTED TO INCREASE TOXIC AMYLOID BETA (ABETA) PRODUCTION VIA ABERRANT CLEAVAGE AT THE BETA-SECRETASE SITE AND THEREBY CAUSE EARLY-ONSET ALZHEIMER'S DISEASE (AD). HOWEVER, THE UNDERLYING MOLECULAR MECHANISMS LEADING TO AD PATHOGENESIS REMAINS LARGELY UNKNOWN. PREVIOUSLY, OUR TRANSCRIPTOME SEQUENCE ANALYSES REVEALED GLOBAL EXPRESSIONAL MODIFICATIONS OF OVER 600 GENES IN APP-SWEDISH MUTANT-EXPRESSING H4 (H4-SW) CELLS COMPARED TO WILD TYPE H4 CELLS. INSULIN-LIKE GROWTH FACTOR BINDING PROTEIN 3 (IGFBP3) IS ONE GENE THAT SHOWED SIGNIFICANTLY DECREASED MRNA EXPRESSION IN H4-SW CELLS. IN THIS STUDY, WE INVESTIGATED THE FUNCTIONAL ROLE OF IGFBP3 IN AD PATHOGENESIS AND ELUCIDATED THE MECHANISMS REGULATING ITS EXPRESSION. WE OBSERVED DECREASED IGFBP3 EXPRESSION IN THE H4-SW CELL LINE AS WELL AS THE HIPPOCAMPUS OF AD MODEL TRANSGENIC MICE. TREATMENT WITH EXOGENOUS IGFBP3 PROTEIN INHIBITED ABETA1-42- INDUCED CELL DEATH AND CASPASE-3 ACTIVITY, WHEREAS SIRNA-MEDIATED SUPPRESSION OF IGFBP3 EXPRESSION INDUCED CELL DEATH AND CASPASE-3 CLEAVAGE. IN PRIMARY HIPPOCAMPAL NEURONS, ADMINISTRATION OF IGFBP3 PROTEIN BLOCKED APOPTOTIC CELL DEATH DUE TO ABETA1-42 TOXICITY. THESE DATA IMPLICATE A PROTECTIVE ROLE FOR IGFBP3 AGAINST ABETA1-42-MEDIATED APOPTOSIS. NEXT, WE INVESTIGATED THE REGULATORY MECHANISMS OF IGFBP3 EXPRESSION IN AD PATHOGENESIS. WE OBSERVED ABNORMAL IGFBP3 HYPERMETHYLATION WITHIN THE PROMOTER CPG ISLAND IN H4-SW CELLS. TREATMENT WITH THE DNA METHYLTRANSFERASE INHIBITOR 5-AZA-2'-DEOXYCYTIDINE RESTORED IGFBP3 EXPRESSION AT BOTH THE MRNA AND PROTEIN LEVELS. CHRONIC EXPOSURE TO ABETA1-42 INDUCED IGFBP3 HYPERMETHYLATION AT CPGS, PARTICULARLY AT LOCI -164 AND -173, AND SUBSEQUENTLY SUPPRESSED IGFBP3 EXPRESSION. THEREFORE, WE DEMONSTRATE THAT EXPRESSION OF ANTI-APOPTOTIC IGFBP3 IS REGULATED BY EPIGENETIC DNA METHYLATION, SUGGESTING A MECHANISM THAT CONTRIBUTES TO AD PATHOGENESIS. 2014 17 2887 25 GADD45A TRANSCRIPTIONAL INDUCTION ELICITED BY THE AURORA KINASE INHIBITOR MK-0457 IN BCR-ABL-EXPRESSING CELLS IS DRIVEN BY OCT-1 TRANSCRIPTION FACTOR. THE ADVANTAGE OF AURORA KINASE (AK) INHIBITORS IN CHRONIC MYELOID LEUKEMIA (CML) THERAPY MOSTLY ARISES FROM "OFF-TARGET" EFFECTS ON TYROSINE KINASE (TK) ACTIVITY OF WILD TYPE (WT) OR MUTATED BCR-ABL PROTEINS WHICH DRIVE THE DISEASE RESISTANCE TO IMATINIB (IM). WE PROVED THAT THE AK INHIBITOR MK-0457 INDUCES THE GROWTH ARREST DNA DAMAGE-INDUCIBLE (GADD) 45A THROUGH RECRUITMENT OF OCTAMER-BINDING (OCT)-1 TRANSCRIPTION FACTOR AT A CRITICAL PROMOTER REGION FOR GENE TRANSCRIPTION AND COVALENT MODIFICATIONS OF HISTONE H3 (LYSINE 14 ACETYLATION, LYSINE 9 DE-METHYLATION). SUCH EPIGENETIC CHROMATIN MODIFICATIONS MAY DEPICT A GENERAL MECHANISM PROMOTING THE RE-ACTIVATION OF TUMOR SUPPRESSOR GENES SILENCED BY BCR-ABL. 2012 18 2080 27 EPIGENETIC DNA METHYLATION OF EBI3 MODULATES HUMAN INTERLEUKIN-35 FORMATION VIA NFKB SIGNALING: A PROMISING THERAPEUTIC OPTION IN ULCERATIVE COLITIS. ULCERATIVE COLITIS (UC), A SEVERE CHRONIC DISEASE WITH UNCLEAR ETIOLOGY THAT IS ASSOCIATED WITH INCREASED RISK FOR COLORECTAL CANCER, IS ACCOMPANIED BY DYSREGULATION OF CYTOKINES. EPSTEIN-BARR VIRUS-INDUCED GENE 3 (EBI3) ENCODES A SUBUNIT IN THE UNIQUE HETERODIMERIC IL-12 CYTOKINE FAMILY OF EITHER PRO- OR ANTI-INFLAMMATORY FUNCTION. AFTER HAVING RECENTLY DEMONSTRATED THAT UPREGULATION OF EBI3 BY HISTONE ACETYLATION ALLEVIATES DISEASE SYMPTOMS IN A DEXTRAN SULFATE SODIUM (DSS)-TREATED MOUSE MODEL OF CHRONIC COLITIS, WE NOW AIMED TO EXAMINE A POSSIBLE FURTHER EPIGENETIC REGULATION OF EBI3 BY DNA METHYLATION UNDER INFLAMMATORY CONDITIONS. TREATMENT WITH THE DNA METHYLTRANSFERASE INHIBITOR (DNMTI) DECITABINE (DAC) AND TNFALPHA LED TO SYNERGISTIC UPREGULATION OF EBI3 IN HUMAN COLON EPITHELIAL CELLS (HCEC). USE OF DIFFERENT SIGNALING PATHWAY INHIBITORS INDICATED NFKAPPAB SIGNALING WAS NECESSARY AND PROPORTIONAL TO THE SYNERGISTIC EBI3 INDUCTION. MALDI-TOF/MS AND HPLC-ESI-MS/MS ANALYSIS OF DAC/TNFALPHA-TREATED HCEC IDENTIFIED IL-12P35 AS THE MOST PROBABLE BINDING PARTNER TO FORM A FUNCTIONAL PROTEIN. EBI3/IL-12P35 HETERODIMERS (IL-35) INDUCE THEIR OWN GENE UPREGULATION, SOMETHING THAT WAS INDEED OBSERVED IN HCEC CULTURED WITH MEDIA FROM PREVIOUSLY DAC/TNFALPHA-TREATED HCEC. THESE RESULTS SUGGEST THAT UNDER INFLAMMATORY AND DEMETHYLATING CONDITIONS THE UPREGULATION OF EBI3 RESULTS IN THE FORMATION OF ANTI-INFLAMMATORY IL-35, WHICH MIGHT BE CONSIDERED AS A THERAPEUTIC TARGET IN COLITIS. 2021 19 145 30 ABERRANT DNA METHYLATION STATUS AND MRNA EXPRESSION LEVEL OF SMG1 GENE IN CHRONIC MYELOID LEUKEMIA: A CASE-CONTROL STUDY. OOBJECTIVE: CHRONIC MYELOID LEUKEMIA (CML) IS A MYELOPROLIFERATIVE MALIGNANCY WITH DIFFERENT STAGES. ABERRANT EPIGENETIC MODIFICATIONS, SUCH AS DNA METHYLATION, HAVE BEEN INTRODUCED AS A SIGNATURE FOR DIVERSE CANCERS WHICH ALSO PLAYS A CRUCIAL ROLE IN CML PATHOGENESIS AND DEVELOPMENT. SUPPRESSOR WITH MORPHOGENETIC EFFECT ON GENITALIA (SMG1) GENE RECENTLY HAS BEEN BROUGHT TO THE SPOTLIGHT AS A POTENT TUMOR SUPPRESSOR GENE THAT CAN BE SUPPRESSED BY TUMORS FOR FURTHER PROGRESS. THE PRESENT STUDY AIMS TO INVESTIGATE SMG1 STATUS IN CML PATIENTS. MATERIALS AND METHODS: IN THIS CASE-CONTROL STUDY, PERIPHERAL BLOOD FROM 30 PATIENTS WITH DIFFERENT PHASES OF CML [NEW CASE (N)=10, COMPLETE MOLECULAR REMISSION (CMR)=10, BLASTIC PHASE (BP)=10] AND 10 HEALTHY SUBJECTS WERE COLLECTED. METHYLATION STATUS AND EXPRESSION LEVEL OF SMG1 GENE PROMOTER WAS ASSESSED BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION (MSP) AND QUANTITATIVE REVERSE-TRANSCRIPTION PCR, RESPECTIVELY. RESULTS: MSP RESULTS OF SMG1 GENE PROMOTOR IN THE NEW CASE GROUP WERE METHYLATED (60% METHYLATED, 30% HEMIMETHYLATED AND 10% UNMETHYLATED). ALL CMR AND CONTROL GROUP PATIENTS WERE UNMETHYLATED IN THE SMG1 GENE PROMOTER. IN THE BP GROUP, METHYLATED SMG1 PROMOTER WAS SEEN (50% OF PATIENTS HAD A METHYLATED STATUS AND 50% HAD HEMIMETHYLATED STATUS). IN COMPARISON WITH THE HEALTHY SUBJECTS, EXPRESSION LEVEL OF SMG1 IN THE NEW CASE GROUP WAS DECREASED (P<0.01); IN THE CMR GROUP AND BP-CML GROUPS, IT WAS INCREASED (P<0.05). NO SIGNIFICANT CORRELATION BETWEEN PATIENTS' HEMATOLOGICAL FEATURES AND SMG1 METHYLATION WAS SEEN. CONCLUSION: OUR RESULTS DEMONSTRATED THAT ABERRANT METHYLATION OF SMG1 OCCURRED IN CML PATIENTS AND IT HAD A SIGNIFICANT ASSOCIATION WITH SMG1 EXPRESSION. SMG1 GENE PROMOTER SHOWED DIVERSE METHYLATED STATUS AND SUBSEQUENT EXPRESSION LEVELS IN DIFFERENT PHASES OF CML. THESE FINDINGS SUGGESTED POSSIBLE PARTICIPATION OF SMG1 SUPPRESSION IN THE CML PATHOGENESIS. 2022 20 829 29 CHARACTERIZATION OF P190-BCR-ABL CHRONIC MYELOID LEUKEMIA REVEALS SPECIFIC SIGNALING PATHWAYS AND THERAPEUTIC TARGETS. THE ONCOGENIC PROTEIN BCR-ABL HAS TWO MAJOR ISOFORMS, P190(BCR-ABL) AND P210(BCR-ABL). WHILE P210(BCR-ABL) IS THE HALLMARK OF CHRONIC MYELOID LEUKEMIA (CML), P190(BCR-ABL) OCCURS IN THE MAJORITY OF PHILADELPHIA-POSITIVE ACUTE LYMPHOBLASTIC LEUKEMIA (PH + ALL) PATIENTS. IN CML, P190(BCR-ABL) OCCURS IN A MINORITY OF PATIENTS ASSOCIATING WITH DISTINCT HEMATOLOGICAL FEATURES AND INFERIOR OUTCOMES, YET THE PATHOGENIC ROLE OF P190(BCR-ABL) AND POTENTIAL TARGETING THERAPIES ARE LARGELY UNCHARACTERIZED. WE EMPLOYED NEXT GENERATION SEQUENCING, PHOSPHO-PROTEOMIC PROFILING, AND DRUG SENSITIVITY TESTING TO CHARACTERIZE P190(BCR-ABL) IN CML AND HEMATOPOIETIC PROGENITOR CELL LINE MODELS (BA/F3 AND HPC-LSK). P190(BCR-ABL) CML PATIENTS DEMONSTRATED POOR RESPONSE TO IMATINIB AND FREQUENT MUTATIONS IN EPIGENETIC MODIFIERS GENES. IN CONTRAST WITH P210(BCR-ABL), P190(BCR-ABL) EXHIBITED SPECIFIC TRANSCRIPTIONAL UPREGULATION OF INTERFERON, INTERLEUKIN-1 RECEPTOR, AND P53 SIGNALING PATHWAYS, ASSOCIATED WITH HYPERPHOSPHORYLATION OF RELEVANT SIGNALING MOLECULES INCLUDING JAK1/STAT1 AND PAK1 IN ADDITION TO SRC HYPERPHOSPHORYLATION. COMPARABLE TO P190(BCR-ABL) CML PATIENTS, P190(BCR-ABL) CELL LINES DEMONSTRATED SIMILAR TRANSCRIPTIONAL AND PHOSPHO-SIGNALING SIGNATURES. WITH THE DRUG SENSITIVITY SCREENING WE IDENTIFIED TARGETED DRUGS WITH SPECIFIC ACTIVITY IN P190(BCR-ABL) CELL LINES INCLUDING IAP-, PAK1-, AND SRC INHIBITORS AND GLUCOCORTICOIDS. OUR RESULTS PROVIDE NOVEL INSIGHTS INTO THE MECHANISMS UNDERLYING THE DISTINCT FEATURES OF P190(BCR-ABL) CML AND PROMISING THERAPEUTIC TARGETS FOR THIS HIGH-RISK PATIENT GROUP. 2021