1 1338 160 DESIGN, SYNTHESIS AND BIOLOGICAL EVALUATION OF A PHENYL BUTYRIC ACID DERIVATIVE, N-(4-CHLOROPHENYL)-4-PHENYLBUTANAMIDE: A HDAC6 INHIBITOR WITH ANTI-PROLIFERATIVE ACTIVITY ON CERVIX CANCER AND LEUKEMIA CELLS. BACKGROUND: THE EPIGENETIC REGULATION OF GENES IN CANCER COULD BE TARGETED BY INHIBITING HISTONE DEACETYLASE 6 (HDAC6), AN ENZYME INVOLVED IN SEVERAL TYPES OF CANCER SUCH AS LYMPHOMA, LEUKEMIA, OVARIAN CANCER, ETC. OBJECTIVE: THROUGH IN SILICO METHODS, A SET OF PHENYL BUTYRIC ACID DERIVATIVES WITH POSSIBLE HDAC6 INHIBITORY ACTIVITY WERE DESIGNED, RENDERING MONOPHENYLAMIDES AND BIPHENYLAMIDES USING TUBACIN (HDAC6 SELECTIVE INHIBITOR) AS REFERENCE. METHOD: THE TARGET COMPOUNDS WERE SUBMITTED TO THEORETICAL ADMET ANALYSES AND THEIR BINDING PROPERTIES ON DIFFERENT HDAC6 CONFORMERS WERE EVALUATED THROUGH DOCKING CALCULATIONS. RESULTS: THESE IN SILICO STUDIES ALLOWED US TO IDENTIFY A COMPOUND NAMED B-R2B. IN ORDER TO HAVE MORE INFORMATION ABOUT THE B-R2B BINDING RECOGNITION PROPERTIES ON HDAC6, THE B-R2B-HDAC6 COMPLEX WAS SUBMITTED THROUGH 100 NS-LONG MOLECULAR DYNAMICS (MD) SIMULATION COUPLED TO MMGBSA APPROACH, REVEALING THAT B-R2B IS LOCATED AT THE ENTRANCE OF HDAC6 ACTIVE POCKET, BLOCKING THE PASSAGE OF THE SUBSTRATE WITHOUT REACHING THE HDAC6 BINDING SITE. BASED ON THESE RESULTS, B-R2B WAS SYNTHESIZED, CHARACTERIZED AND BIOLOGICALLY TESTED. THE HDAC6 FLUOROMETRIC DRUG DISCOVERY KIT FLUOR-DE-LYS (ENZO LIFE SCIENCES INC.) WAS USED TO DETERMINE THE HDAC6 HUMAN INHIBITORY ACTIVITY (IC50 VALUE) OF B-R2B COMPOUND. IN ADDITION, B-R2B SHOW IC50 VALUES ON CANCER CELL LINES (HELA; IC50 = 72.6 MICROM), ACUTE MYELOID LEUKEMIA (THP-1; IC50 = 16.5 MICROM), HUMAN MAST LEUKEMIA (HMC; IC50 = 79.29 MICROM) AND CHRONIC MYELOGENOUS LEUKEMIA (KASUMI; IC50 = 101 MICROM). CONCLUSION: THESE RESULTS SHOW THAT B-R2B IS A HDAC6 INHIBITOR, SPECIFICALLY A NON-COMPETITIVE TYPE IN A SIMILAR WAY THAT TUBACIN DOES, ACCORDING TO MD SIMULATIONS. 2017 2 6586 32 TUBASTATIN, A SELECTIVE HISTONE DEACETYLASE 6 INHIBITOR SHOWS ANTI-INFLAMMATORY AND ANTI-RHEUMATIC EFFECTS. EPIGENETIC MODIFICATIONS REPRESENT A PROMISING NEW APPROACH TO MODULATE CELL FUNCTIONS AS OBSERVED IN AUTOIMMUNE DISEASES. EMERGING EVIDENCE SUGGESTS THE UTILITY OF HDAC INHIBITORS IN THE TREATMENT OF CHRONIC IMMUNE AND INFLAMMATORY DISORDERS. HOWEVER, CLASS AND ISOFORM SELECTIVE INHIBITION OF HDAC IS CURRENTLY FAVORED AS IT LIMITS THE TOXICITY THAT HAS BEEN OBSERVED WITH PAN-HDAC INHIBITORS. HDAC6, A MEMBER OF THE HDAC FAMILY, WHOSE MAJOR SUBSTRATE IS ALPHA-TUBULIN, IS BEING INCREASINGLY IMPLICATED IN THE PATHOGENESIS OF INFLAMMATORY DISORDERS. THE PRESENT STUDY WAS CARRIED OUT TO STUDY THE POTENTIAL ANTI-INFLAMMATORY AND ANTI-RHEUMATIC EFFECTS OF HDAC6 SELECTIVE INHIBITOR TUBASTATIN. TUBASTATIN, A POTENT HUMAN HDAC6 INHIBITOR WITH AN IC50 OF 11 NM SHOWED SIGNIFICANT INHIBITION OF TNF-ALPHA AND IL-6 IN LPS STIMULATED HUMAN THP-1 MACROPHAGES WITH AN IC50 OF 272 NM AND 712 NM RESPECTIVELY. ADDITIONALLY, TUBASTATIN INHIBITED NITRIC OXIDE (NO) SECRETION IN MURINE RAW 264.7 MACROPHAGES DOSE DEPENDENTLY WITH AN IC50 OF 4.2 MUM AND INDUCED ALPHA-TUBULIN HYPERACETYLATION CORRESPONDING TO HDAC6 INHIBITION IN THP-1 CELLS WITHOUT AFFECTING THE CELL VIABILITY. TUBASTATIN SHOWED SIGNIFICANT INHIBITION OF PAW VOLUME AT 30 MG/KG I.P. IN A FREUND'S COMPLETE ADJUVANT (FCA) INDUCED ANIMAL MODEL OF INFLAMMATION. THE DISEASE MODIFYING ACTIVITY OF TUBASTATIN WAS ALSO EVIDENT IN COLLAGEN INDUCED ARTHRITIS DBA1 MOUSE MODEL AT 30 MG/KG I.P. THE SIGNIFICANT ATTENUATION OF CLINICAL SCORES (~70%) BY TUBASTATIN WAS CONFIRMED HISTOPATHOLOGICALLY AND WAS FOUND COMPARABLE TO DEXAMETHASONE (~90% INHIBITION OF CLINICAL SCORES). TUBASTATIN SHOWED SIGNIFICANT INHIBITION OF IL-6 IN PAW TISSUES OF ARTHRITIC MICE. THE PRESENT WORK HAS DEMONSTRATED ANTI-INFLAMMATORY AND ANTIRHEUMATIC EFFECTS OF A SELECTIVE HDAC6 INHIBITOR TUBASTATIN. 2013 3 2761 33 EXPRESSION OF TESTIS-SPECIFIC GENES, TEX101 AND ODF4, IN CHRONIC MYELOID LEUKEMIA AND EVALUATION OF TEX101 IMMUNOGENICITY. BACKGROUND AND OBJECTIVES: CANCER-TESTIS (CT) ANTIGENS ARE A GROUP OF ANTIGENS WITH A RESTRICTED EXPRESSION IN NORMAL TISSUES, EXCEPT TESTIS, AND THEY HAVE ABERRANT EXPRESSION IN DIFFERENT TUMORS. THIS PATTERN OF EXPRESSION HAS MADE THEM PROMISING TARGETS FOR IMMUNOTHERAPY AND CANCER DETECTION. OUR AIM WAS TO FIND NEW MEMBERS OF THIS GROUP THAT MIGHT BE USEFUL AS MARKERS IN THE DETECTION OF CANCER AND IMMUNOTHERAPY. DESIGN AND SETTING: A DESCRIPTIVE STUDY CONDUCTED IN REFERRAL CENTERS OF TEHRAN UNIVERSITY OF MEDICAL SCIENCE FROM JANUARY 2008 TO JANUARY 2009. PATIENTS AND METHODS: WE ANALYZED THE EXPRESSION OF TWO TESTIS-SPECIFIC GENES NAMED ODF4 (OUTER DENSE FIBER OF SPERM TAILS 4) AND TEX101 (TESTIS EXPRESSED 101) IN 20 CHRONIC MYELOID LEUKEMIA (CML) AND 20 NORMAL SAMPLES BY REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION AND SEQUENCING. IMMUNOGENICITY OF TEX101 WAS EVALUATED BY MEANS OF ENZYME-LINKED IMMUNOSORBENT ASSAY. RESULTS: THESE TWO GENES WERE EXPRESSED IN 30% OF CML PATIENTS BUT NOT IN ANY OF THE HEALTHY DONORS. HUMORAL RESPONSE AGAINST TEX101 WAS NOT DETECTED IN ANY SAMPLES. CONCLUSIONS: TEX101 AND ODF4 ARE CT GENES USEFUL FOR DETECTION OF CML. UNLIKE MANY CT GENES, OVEREXPRESSION OF TEX101 WAS NOT SHOWN TO INDUCE IMMUNOLOGIC RESPONSES IN THESE SAMPLES. ACCORDING TO THE PREVIOUS STUDIES, OVEREXPRESSION OF TEX101 LEADS TO SUPPRESSION OF CANCER INVASION AND METASTASIS; THUS, THE INDUCTION OF THE EXPRESSION OF TEX101 IN CANCER BY EPIGENETIC MECHANISMS MAY BE A TREATMENT STRATEGY. 2012 4 3198 35 HDAC-LINKED "PROLIFERATIVE" MIRNA EXPRESSION PATTERN IN PANCREATIC NEUROENDOCRINE TUMORS. EPIGENETIC FACTORS ARE ESSENTIALLY INVOLVED IN CARCINOGENESIS, TUMOR PROMOTION, AND CHEMORESISTANCE. TWO EPIGENETIC KEY PLAYERS ARE MIRNAS AND HISTONE DEACETYLASES (HDACS). AS PREVIOUSLY SHOWN BY OWN THEORETICAL DATABANK ANALYSIS, THE CROSSTALK BETWEEN MIRNAS AND HDACS IS RELEVANT IN DIFFERENT HUMAN CHRONIC DISEASES AND CANCEROGENIC PATHWAYS. WE AIMED TO INVESTIGATE A POTENTIAL CONNECTION BETWEEN THE EXPRESSION OF A WELL-DEFINED SUBSET OF "PROLIFERATION-ASSOCIATED" MIRNAS AND THE EXPRESSION OF HDACS AS WELL AS CLINICAL PARAMETERS IN PANCREATIC NEUROENDOCRINE TUMORS (PNETS). MATERIALS AND METHODS: EXPRESSION LEVELS OF MIRNA132-3P, MIRNA145-5P, MIRNA183-5P, MIRNA34A-5P, AND MIRNA449A IN 57 PNETS RESECTED BETWEEN 1997 AND 2015 WERE MEASURED AND LINKED TO THE IMMUNOHISTOCHEMICAL EXPRESSION PATTERN OF MEMBERS OF THE FOUR HDAC CLASSES ON HUMAN TISSUE MICROARRAYS. ALL PNET CASES WERE CLINICALLY AND PATHOLOGICALLY CHARACTERIZED ACCORDING TO PUBLISHED GUIDELINES. CORRELATION ANALYSIS REVEALED A SIGNIFICANT ASSOCIATION BETWEEN EXPRESSION OF SPECIFIC MIRNAS AND TWO MEMBERS OF THE HDAC FAMILY (HDAC3 AND HDAC4). ADDITIONALLY, A LINKAGE BETWEEN MIRNA EXPRESSION AND CLINICO-PATHOLOGICAL PARAMETERS LIKE GRADING, TNM-STAGING, AND HORMONE ACTIVITY WAS FOUND. MOREOVER, OVERALL AND DISEASE-FREE SURVIVAL IS STATISTICALLY CORRELATED WITH THE EXPRESSION OF THE INVESTIGATED MIRNAS. OVERALL, WE DEMONSTRATED THAT SPECIFIC MIRNAS COULD BE LINKED TO HDAC EXPRESSION IN PNETS. ESPECIALLY MIRNA449A (ASSOCIATED WITH HDAC3/4) SEEMS TO PLAY AN IMPORTANT ROLE IN PNET PROLIFERATION AND COULD BE A POTENTIAL PROGNOSTIC FACTOR FOR POOR SURVIVAL. THESE FIRST DATA COULD HELP, TO IMPROVE OUR KNOWLEDGE OF THE COMPLEX INTERACTIONS OF THE EPIGENETIC DRIVERS IN PNETS FOR FURTHER THERAPEUTIC APPROACHES. 2018 5 2733 46 EXPLORING THE POTENTIAL OF PHYTOCOMPOUNDS FOR TARGETING EPIGENETIC MECHANISMS IN RHEUMATOID ARTHRITIS: AN IN SILICO STUDY USING SIMILARITY INDEXING. FINDING STRUCTURALLY SIMILAR COMPOUNDS IN COMPOUND DATABASES IS HIGHLY EFFICIENT AND IS WIDELY USED IN PRESENT-DAY DRUG DISCOVERY METHODOLOGY. THE MOST-TRUSTED AND -FOLLOWED SIMILARITY INDEXING METHOD IS TANIMOTO SIMILARITY INDEXING. EPIGENETIC PROTEINS LIKE HISTONE DEACETYLASES (HDACS) INHIBITORS ARE TRADITIONALLY USED TO TARGET CANCER, BUT HAVE ONLY BEEN INVESTIGATED VERY RECENTLY FOR THEIR POSSIBLE EFFECTIVENESS AGAINST RHEUMATOID ARTHRITIS (RA). THE SYNTHETIC DRUGS THAT HAVE BEEN IDENTIFIED AND USED FOR THE INHIBITION OF HDACS INCLUDE SAHA, WHICH IS BEING USED TO INHIBIT THE ACTIVITY OF HDACS OF DIFFERENT CLASSES. SAHA WAS CHOSEN AS A COMPOUND OF HIGH IMPORTANCE AS IT IS REPORTED TO INHIBIT THE ACTIVITY OF MANY HDAC TYPES. SIMILARITY SEARCHING USING THE UNPD DATABASE AS A REFERENCE IDENTIFIED AGLAITHIODULINE FROM THE AGLAIA LEPTANTHA COMPOUND AS HAVING A ~70% SIMILARITY OF MOLECULAR FINGERPRINTS WITH SAHA, BASED ON THE TANIMOTO INDEXING METHOD USING CHEMMINER. AGLAITHIODULINE IS ABUNDANTLY PRESENT IN THE SHELL AND FRUITS OF A. LEPTANTHA. IN SILICO STUDIES WITH AGLAITHIODULINE WERE CARRIED OUT AGAINST THE HDAC8 PROTEIN TARGET AND SHOWED A BINDING AFFINITY OF -8.5 KCAL MOL. THE COMPLEX WAS FURTHER SUBJECTED TO MOLECULAR DYNAMICS SIMULATION USING GROMACS. THE RMSD, RMSF, COMPACTNESS AND SASA PLOTS OF THE TARGET WITH AGLAITHIODULINE, IN COMPARISON WITH THE CO-CRYSTALLIZED LIGAND (SAHA) SYSTEM, SHOWED A VERY STABLE CONFIGURATION. THE RESULTS OF THE STUDY ARE SUPPORTIVE OF THE USAGE OF A. LEPTANTHA AND A. EDULIS IN INDIAN TRADITIONAL MEDICINE FOR THE TREATMENT OF PAIN-RELATED AILMENTS SIMILAR TO RA. OUR STUDY THEREFORE CALLS FOR FURTHER INVESTIGATION OF A. LEPTANTHA AND A. EDULIS FOR THEIR POTENTIAL USE AGAINST RA BY TARGETING EPIGENETIC CHANGES, USING IN VIVO AND IN VITRO STUDIES. 2023 6 2763 28 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 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 8 1334 30 DEREGULATION OF AIOLOS EXPRESSION IN CHRONIC LYMPHOCYTIC LEUKEMIA IS ASSOCIATED WITH EPIGENETIC MODIFICATIONS. CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IS CHARACTERIZED BY A CLONAL ACCUMULATION OF MATURE NEOPLASTIC B CELLS THAT ARE RESISTANT TO APOPTOSIS. AIOLOS, A MEMBER OF THE IKAROS FAMILY OF ZINC-FINGER TRANSCRIPTION FACTORS, PLAYS AN IMPORTANT ROLE IN THE CONTROL OF MATURE B LYMPHOCYTE DIFFERENTIATION AND MATURATION. IN THIS STUDY, WE SHOWED THAT AIOLOS EXPRESSION IS UP-REGULATED IN B-CLL CELLS. THIS OVEREXPRESSION DOES NOT IMPLICATE ISOFORM IMBALANCE OR DISTURB AIOLOS SUBCELLULAR LOCALIZATION. THE CHROMATIN STATUS AT THE AIOLOS PROMOTER IN CLL IS DEFINED BY THE DEMETHYLATION OF DNA AND AN ENRICHMENT OF EUCHROMATIN ASSOCIATED HISTONE MARKERS, SUCH AS THE DIMETHYLATION OF THE LYSINE 4 ON HISTONE H3. THESE EPIGENETIC MODIFICATIONS SHOULD ALLOW ITS UPSTREAM EFFECTORS, SUCH AS NUCLEAR FACTOR-KAPPAB, CONSTITUTIVELY ACTIVATED IN CLL, TO GAIN ACCESS TO PROMOTER, RESULTING UP-REGULATION OF AIOLOS. TO DETERMINE THE CONSEQUENCES OF AIOLOS DEREGULATION IN CLL, WE ANALYZED THE EFFECTS OF AIOLOS OVEREXPRESSION OR DOWN-REGULATION ON APOPTOSIS. AIOLOS IS INVOLVED IN CELL SURVIVAL BY REGULATING THE EXPRESSION OF SOME BCL-2 FAMILY MEMBERS. OUR RESULTS STRONGLY SUGGEST THAT AIOLOS DEREGULATION BY EPIGENETIC MODIFICATIONS MAY BE A HALLMARK OF CLL. 2011 9 3484 39 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 10 5101 35 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 11 2767 37 EXPRESSION, PROGNOSTIC VALUE, AND FUNCTIONAL MECHANISM OF THE KDM5 FAMILY IN PANCREATIC CANCER. BACKGROUND: THE HISTONE LYSINE DEMETHYLASE KDM5 FAMILY IS AN IMPORTANT EPIGENETIC STATE-MODIFYING ENZYME FAMILY. INCREASING EVIDENCE SUPPORTS THAT EPIGENETIC ABNORMALITIES IN THE KDM5 FAMILY ARE RELATED TO MULTIPLE CANCERS IN HUMANS. HOWEVER, THE ROLE OF THE KDM5 FAMILY IN PANCREATIC CANCER IS NOT CLEAR, AND RELATED RESEARCH IS VERY SCARCE. METHODS: R SOFTWARE, KAPLAN-MEIER PLOTTER, CBIOPORTAL, TIMER, LINKEDOMICS, STRING, METASCAPE, TISIDB, AND THE GSCA LITE ONLINE TOOL WERE UTILIZED FOR BIOINFORMATICS ANALYSIS. RESULTS: KDM5A/B/C WAS SIGNIFICANTLY OVEREXPRESSED IN MANY KINDS OF TUMOR TISSUES, INCLUDING PANCREATIC ADENOCARCINOMA (PAAD), WHILE THE EXPRESSION OF KDM5D WAS SIGNIFICANTLY DOWNREGULATED. THE HIGH EXPRESSION OF KDM5A/B/C WAS RELATED TO POOR CLINICAL FEATURES, SUCH AS WORSE TREATMENT EFFICACY, HIGHER TUMOR GRADE, AND MORE ADVANCED CLINICAL STAGE. PATIENTS WITH A FAMILY HISTORY OF BREAST CANCER AND MELANOMA, HISTORY OF DRINKING OR HISTORY CHRONIC PANCREATITIS WERE MORE LIKELY TO HAVE KDM5A/B/C GENE ABNORMALITIES, WHICH WERE RELATED TO A VARIETY OF ADVERSE CLINICAL FEATURES. THE RESULTS OF GENE ONTOLOGY (GO) AND KYOTO ENCYCLOPEDIA OF GENES AND GENOMES (KEGG) PATHWAY ANALYSES OF THE KDM5 FAMILY AND ITS 800 CO-EXPRESSED GENES SHOWED THAT MANY GENE TERMS RELATED TO CELL PROLIFERATION, MIGRATION AND MANY CARCINOGENIC PATHWAYS. NOTABLY, WE FOUND THAT THE EXPRESSION LEVEL OF KDM5A/B/C WAS POSITIVELY CORRELATED WITH THE EXPRESSION OF MULTIPLE KEY DRIVER GENES SUCH AS KRAS, BRCA1, AND BRCA2 ETC. IN ADDITION, PPI NETWORK ANALYSIS SHOWED KDM5 FAMILY PROTEINS HAVE STRONG INTERACTIONS WITH HISTONE DEACETYLASE FAMILY 1 (HDAC1), WHICH COULD MODIFY THE LYSINES OF HISTONE H3, AND CO-ACT ON MANY PATHWAYS, INCLUDING THE "LONGEVITY-REGULATING PATHWAY" AND "NOTCH SIGNALING PATHWAY". MOREOVER, THE UPREGULATION OF KDM5A/B/C EXPRESSION WAS ASSOCIATED WITH AN INCREASE IN THE INFILTRATION OF B CELLS, CD8(+) T CELLS AND OTHER INFILTRATING IMMUNE LYMPHOCYTES AND THE EXPRESSION LEVELS OF IMMUNE MOLECULES SUCH AS NT5E AND CD274. INTERESTINGLY, THE OVEREXPRESSION OF KDM5A/C WAS ALSO CORELATED WITH REDUCED SENSITIVITY OF PANCREATIC CANCER CELLS TO MANY KINDS OF PANCREATIC CANCER-TARGETING OR CHEMOTHERAPEUTIC DRUGS, INCLUDING AXITINIB AND GEMCITABINE. CONCLUSION: KDM5 FAMILY MEMBERS MAY BE PROGNOSTIC MARKERS AND NEW THERAPEUTIC TARGETS FOR PATIENTS WITH PANCREATIC CANCER. 2022 12 3128 37 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 13 671 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 14 1806 28 EFFECT OF TRANSCRIPTION INHIBITION AND GENERATION OF SUPPRESSIVE VIRAL NON-CODING RNAS. BACKGROUND: HIV-1 PATIENTS RECEIVING COMBINATION ANTIRETROVIRAL THERAPY (CART) SURVIVE INFECTION BUT REQUIRE LIFE-LONG ADHERENCE AT HIGH EXPENSE. IN CHRONIC CART-TREATED PATIENTS WITH UNDETECTABLE VIRAL TITERS, CELL-ASSOCIATED VIRAL RNA IS STILL DETECTABLE, POINTING TO LOW-LEVEL VIRAL TRANSCRIPTIONAL LEAKINESS. TO DATE, THERE ARE NO FDA-APPROVED DRUGS AGAINST HIV-1 TRANSCRIPTION. WE HAVE PREVIOUSLY SHOWN THAT F07#13, A THIRD GENERATION TAT PEPTIDE MIMETIC WITH COMPETITIVE ACTIVITY AGAINST CDK9/T1-TAT BINDING SITES, INHIBITS HIV-1 TRANSCRIPTION IN VITRO AND IN VIVO. RESULTS: HERE, WE DEMONSTRATE THAT INCREASING CONCENTRATIONS OF F07#13 (0.01, 0.1, 1 MICROM) CAUSE A DECREASE IN TAT LEVELS IN A DOSE-DEPENDENT MANNER BY INHIBITING THE CDK9/T1-TAT COMPLEX FORMATION AND SUBSEQUENT UBIQUITIN-MEDIATED TAT SEQUESTRATION AND DEGRADATION. OUR DATA INDICATE THAT COMPLEXES I AND IV CONTAIN DISTINCT PATTERNS OF UBIQUITINATED TAT AND THAT TRANSCRIPTIONAL INHIBITION INDUCED BY F07#13 CAUSES AN OVERALL REDUCTION IN TAT LEVELS. THIS REDUCTION MAY BE TRIGGERED BY F07#13 BUT ULTIMATELY IS MEDIATED BY TAR-GAG VIRAL RNAS THAT BIND SUPPRESSIVE TRANSCRIPTION FACTORS (SIMILAR TO 7SK, NRON, HOTAIR, AND XIST LNCRNAS) TO ENHANCE TRANSCRIPTIONAL GENE SILENCING AND LATENCY. THESE RNAS COMPLEX WITH PRC2, SIN3A, AND CUL4B, RESULTING IN EPIGENETIC MODIFICATIONS. FINALLY, WE OBSERVED AN F07#13-MEDIATED DECREASE OF VIRAL BURDEN BY TARGETING THE R REGION OF THE LONG TERMINAL REPEAT (HIV-1 PROMOTER REGION, LTR), PROMOTING BOTH PAUSED POLYMERASES AND INCREASED EFFICIENCY OF CRISPR/CAS9 EDITING IN INFECTED CELLS. THIS IMPLIES THAT GENE EDITING MAY BE BEST PERFORMED UNDER A REPRESSED TRANSCRIPTIONAL STATE. CONCLUSIONS: COLLECTIVELY, OUR RESULTS INDICATE THAT F07#13, WHICH CAN TERMINATE RNA POLYMERASE II AT DISTINCT SITES, CAN GENERATE SCAFFOLD RNAS, WHICH MAY ASSEMBLE INTO SPECIFIC SETS OF "RNA MACHINES" THAT CONTRIBUTE TO GENE REGULATION. IT REMAINS TO BE SEEN WHETHER THESE EFFECTS CAN ALSO BE SEEN IN VARIOUS CLADES THAT HAVE VARYING PROMOTER STRENGTH, MUTANT LTRS, AND IN PATIENT SAMPLES. 2019 15 4400 34 MODULATION OF HEPATIC STELLATE CELLS BY MUTAFLOR((R)) PROBIOTIC IN NON-ALCOHOLIC FATTY LIVER DISEASE MANAGEMENT. BACKGROUND: NAFLD AND NASH ARE EMERGING AS PRIMARY CAUSES OF CHRONIC LIVER DISEASE, INDICATING A NEED FOR AN EFFECTIVE TREATMENT. MUTAFLOR(R) PROBIOTIC, A MICROBIAL TREATMENT OF INTEREST, WAS EFFECTIVE IN SUSTAINING REMISSION IN ULCERATIVE COLITIS PATIENTS. OBJECTIVE: TO CONSTRUCT A GENETIC-EPIGENETIC NETWORK LINKED TO HSC SIGNALING AS A MODULATOR OF NAFLD/NASH PATHOGENESIS, THEN ASSESS THE EFFECTS OF MUTAFLOR((R)) ON THIS NETWORK. METHODS: FIRST, IN SILICO ANALYSIS WAS USED TO CONSTRUCT A GENETIC-EPIGENETIC NETWORK LINKED TO HSC SIGNALING. SECOND, AN INVESTIGATION USING RATS, INCLUDING HFHSD INDUCED NASH AND MUTAFLOR((R)) TREATED ANIMALS, WAS DESIGNED. EXPERIMENTAL PROCEDURES INCLUDED BIOCHEMICAL AND HISTOPATHOLOGIC ANALYSIS OF RAT BLOOD AND LIVER SAMPLES. AT THE MOLECULAR LEVEL, THE EXPRESSION OF GENETIC (FOXA2, TEAD2, AND LATS2 MRNAS) AND EPIGENETIC (MIR-650, RPARP AS-1 LNCRNA) NETWORK WAS MEASURED BY REAL-TIME PCR. PCR RESULTS WERE VALIDATED WITH IMMUNOHISTOCHEMISTRY (ALPHA-SMA AND LATS2). TARGET EFFECTOR PROTEINS, IL-6 AND TGF-BETA, WERE ESTIMATED BY ELISA. RESULTS: MUTAFLOR((R)) ADMINISTRATION MINIMIZED BIOCHEMICAL AND HISTOPATHOLOGIC ALTERATIONS CAUSED BY NAFLD/NASH. HSC ACTIVATION AND EXPRESSION OF PROFIBROGENIC IL-6 AND TGF-BETA EFFECTOR PROTEINS WERE REDUCED VIA INHIBITION OF HEDGEHOG AND HIPPO PATHWAYS. PATHWAYS MAY HAVE BEEN INHIBITED THROUGH UPREGULATION OF RPARP AS-1 LNCRNA WHICH IN TURN DOWNREGULATED THE EXPRESSION OF MIR-650, FOXA2 MRNA AND TEAD2 MRNA AND UPREGULATED LATS2 MRNA EXPRESSION. CONCLUSION: MUTAFLOR((R)) MAY SLOW THE PROGRESSION OF NAFLD/NASH BY MODULATING A GENETIC-EPIGENETIC NETWORK LINKED TO HSC SIGNALING. THE PROBIOTIC MAY BE A USEFUL MODALITY FOR THE PREVENTION AND TREATMENT OF NAFLD/NASH. 2022 16 493 26 ASSESSMENT OF P53 AND ATM FUNCTIONALITY IN CHRONIC LYMPHOCYTIC LEUKEMIA BY MULTIPLEX LIGATION-DEPENDENT PROBE AMPLIFICATION. THE ATM-P53 DNA-DAMAGE RESPONSE (DDR) PATHWAY HAS A CRUCIAL ROLE IN CHEMORESISTANCE IN CLL, AS INDICATED BY THE ADVERSE PROGNOSTIC IMPACT OF GENETIC ABERRATIONS OF TP53 AND ATM. IDENTIFYING AND DISTINGUISHING TP53 AND ATM FUNCTIONAL DEFECTS HAS BECOME RELEVANT AS EPIGENETIC AND POSTTRANSCRIPTIONAL DYSREGULATION OF THE ATM/P53 AXIS IS INCREASINGLY BEING RECOGNIZED AS THE UNDERLYING CAUSE OF CHEMORESISTANCE. ALSO, SPECIFIC TREATMENTS SENSITIZING TP53- OR ATM-DEFICIENT CLL CELLS ARE EMERGING. WE THEREFORE DEVELOPED A NEW ATM-P53 FUNCTIONAL ASSAY WITH THE AIM TO (I) IDENTIFY AND (II) DISTINGUISH ABNORMALITIES OF TP53 VERSUS ATM AND (III) ENABLE THE IDENTIFICATION OF ADDITIONAL DEFECTS IN THE ATM-P53 PATHWAY. REVERSED TRANSCRIPTASE MULTIPLEX LIGATION-DEPENDENT PROBE AMPLIFICATION (RT-MLPA) WAS USED TO MEASURE ATM AND/OR P53-DEPENDENT GENES AT THE RNA LEVEL FOLLOWING DNA DAMAGE USING IRRADIATION. HERE, WE SHOWED THAT THIS ASSAY IS ABLE TO IDENTIFY AND DISTINGUISH THREE SUBGROUPS OF CLL TUMORS (I.E., TP53-DEFECTIVE, ATM-DEFECTIVE AND WT) AND IS ALSO ABLE TO DETECT ADDITIONAL SAMPLES WITH A DEFECTIVE DDR, WITHOUT MOLECULAR ABERRATIONS IN TP53 AND/OR ATM. THESE FINDINGS MAKE THE ATM-P53 RT-MLPA FUNCTIONAL ASSAY A PROMISING PROGNOSTIC TOOL FOR PREDICTING TREATMENT RESPONSES IN CLL. 2015 17 5775 33 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 18 3049 22 GENOME-WIDE ANALYSIS REVEALS ZINC TRANSPORTER ZIP9 REGULATED BY DNA METHYLATION PROMOTES RADIATION-INDUCED SKIN FIBROSIS VIA THE TGF-BETA SIGNALING PATHWAY. RADIATION-INDUCED SKIN FIBROSIS IS A DETRIMENTAL AND CHRONIC DISORDER THAT OCCURS AFTER RADIATION EXPOSURE. DNA METHYLATION HAS BEEN CHARACTERIZED AS AN IMPORTANT REGULATORY MECHANISM OF MULTIPLE PATHOLOGICAL PROCESSES. IN THIS STUDY, WE COMPARED THE GENOME-WIDE DNA METHYLATION STATUS IN RADIATION-INDUCED FIBROTIC SKIN AND ADJACENT NORMAL TISSUES OF RATS BY METHYLATED DNA IMMUNOPRECIPITATION SEQUENCING. RADIATION-INDUCED FIBROTIC SKIN SHOWED DIFFERENTIALLY METHYLATED REGIONS ASSOCIATED WITH 3,650 PROTEIN-CODING GENES, 72 MICRORNAS, 5,836 LONG NONCODING RNAS AND 3 PIWI-INTERACTING RNAS. BY INTEGRATING THE MRNA AND METHYLATION PROFILES, THE ZINC TRANSPORTER SLC39A9/ZIP9 WAS INVESTIGATED IN GREATER DETAIL. THE PROTEIN LEVEL OF ZIP9 WAS INCREASED IN IRRADIATED SKIN TISSUES OF HUMANS, MONKEYS, AND RATS, ESPECIALLY IN RADIOGENIC FIBROTIC SKIN TISSUES. RADIATION INDUCED THE DEMETHYLATION OF A CPG DINUCLEOTIDE IN EXON 1 OF ZIP9 THAT RESULTED IN RECRUITMENT OF THE TRANSCRIPTIONAL FACTOR SP1 AND INCREASED ZIP9 EXPRESSION. OVEREXPRESSION OF ZIP9 RESULTED IN ACTIVATION OF THE PROFIBROTIC TRANSFORMING GROWTH FACTOR-BETA SIGNALING PATHWAY THROUGH PROTEIN KINASE B IN HUMAN FIBROBLASTS. IN ADDITION, RADIATION-INDUCED SKIN FIBROSIS WAS ASSOCIATED WITH INCREASED ZINC ACCUMULATION. THE ZINC CHELATOR N,N,N',N'-TETRAKIS(2-PYRIDYLMETHYL)-1,2-ETHYLENEDIAMINE ABROGATED ZIP9-INDUCED ACTIVATION OF THE TRANSFORMING GROWTH FACTOR-BETA SIGNALING PATHWAY AND ATTENUATED RADIATION-INDUCED SKIN FIBROSIS IN A RAT MODEL. IN SUMMARY, OUR FINDINGS ILLUSTRATE EPIGENETIC REGULATION OF ZIP9 AND ITS CRITICAL ROLE IN PROMOTING RADIATION-INDUCED SKIN FIBROSIS. 2020 19 3718 31 INHIBITION OF BCL6B PROMOTES GASTRIC CANCER BY AMPLIFYING INFLAMMATION IN MICE. BACKGROUND: CHRONIC GASTRITIS HAS BEEN DEMONSTRATED TO BE A KEY CAUSE OF GASTRIC CANCER (GC), AND CONTROL OF GASTRIC INFLAMMATION IS REGARDED AS AN EFFECTIVE TREATMENT FOR THE CLINICAL PREVENTION OF GASTRIC CARCINOGENESIS. HOWEVER, THERE REMAINS AN UNMET NEED TO IDENTIFY THE DOMINANT REGULATORS OF GASTRIC ONCOGENESIS-ASSOCIATED INFLAMMATION IN VIVO. METHODS: THE MOUSE MODEL FOR THE STUDY OF INFLAMMATION-ASSOCIATED GC WAS INDUCED BY BENZO[A]PYRENE (BAP) INTRAGASTRIC ADMINISTRATION IN BCL6B(-/-) AND WILDTYPE MICE ON A C57BL/6 BACKGROUND. 5-AZA-2'-DEOXYCYTIDINE (5-AZA), THE DEMETHYLATION DRUG, WAS INTRAPERITONEALLY INJECTED TO RESTORE BCL6B EXPRESSION. HUMAN GC TISSUE ARRAY WAS USED TO ANALYSE PATIENT SURVIVAL BASED ON BCL6B AND CD3 PROTEIN EXPRESSION. RESULTS: BCL6B WAS GRADUALLY DOWNREGULATED BY ITS OWN PROMOTER HYPERMETHYLATION IN PARALLEL TO AN INCREASING INFLAMMATORY RESPONSE DURING THE PROGRESSION OF BAP-INDUCED GASTRIC CARCINOGENESIS IN MICE. MOREOVER, KNOCKOUT OF BCL6B DRAMATICALLY WORSENED THE SEVERITY OF GASTRIC CANCER AND AGGRAVATED THE INFLAMMATORY RESPONSE IN THE BAP-INDUCED MICE GC MODEL. RE-ACTIVATION OF BCL6B BY 5-AZA IMPEDED INFLAMMATORY AMPLIFICATION AND BAP-INDUCED GC DEVELOPMENT, PROLONGING SURVIVAL TIME IN WILDTYPE MICE, WHEREAS NO NOTABLE CURATIVE EFFECT OCCURRED IN BCL6B(-/-) MICE WITH 5-AZA TREATMENT. FINALLY, SIGNIFICANT NEGATIVE CORRELATIONS WERE DETECTED BETWEEN THE MRNA LEVELS OF BCL6B AND INFLAMMATORY CYTOKINES IN HUMAN GC TISSUES; PATIENTS HARBOURING BCL6B-NEGETIVE AND SEVERE-INFLAMMATION GC TUMOURS WERE FOUND TO EXHIBIT THE SHORTEST SURVIVAL TIME. CONCLUSIONS: EPIGENETIC INACTIVATION OF BCL6B PROMOTES GASTRIC CANCER THROUGH AMPLIFICATION OF THE GASTRIC INFLAMMATORY RESPONSE IN VIVO AND OFFERS A NEW APPROACH FOR GC TREATMENT AND REGENERATIVE MEDICINE. 2019 20 1843 32 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