1 2793 101 FATE-MAPPING OF GM-CSF EXPRESSION IDENTIFIES A DISCRETE SUBSET OF INFLAMMATION-DRIVING T HELPER CELLS REGULATED BY CYTOKINES IL-23 AND IL-1BETA. PATHOGENIC LYMPHOCYTES INITIATE THE DEVELOPMENT OF CHRONIC INFLAMMATORY DISEASES. THE CYTOKINE GRANULOCYTE-MACROPHAGE COLONY-STIMULATING FACTOR (GM-CSF) (ENCODED BY CSF2) IS A KEY COMMUNICATOR BETWEEN PATHOGENIC LYMPHOCYTES AND TISSUE-INVADING INFLAMMATORY PHAGOCYTES. HOWEVER, THE MOLECULAR PROPERTIES OF GM-CSF-PRODUCING CELLS AND THE MODE OF CSF2 REGULATION IN VIVO REMAIN UNCLEAR. TO SYSTEMATICALLY STUDY AND MANIPULATE GM-CSF(+) CELLS AND THEIR PROGENY IN VIVO, WE GENERATED A FATE-MAP AND REPORTER OF GM-CSF EXPRESSION MOUSE STRAIN (FROG). WE MAPPED THE PHENOTYPIC AND FUNCTIONAL PROFILE OF AUTO-AGGRESSIVE T HELPER (TH) CELLS DURING NEUROINFLAMMATION AND IDENTIFIED THE SIGNATURE AND PATHOGENIC MEMORY OF A DISCRETE ENCEPHALITOGENIC TH SUBSET. THESE CELLS REQUIRED INTERLEUKIN-23 RECEPTOR (IL-23R) AND IL-1R BUT NOT IL-6R SIGNALING FOR THEIR MAINTENANCE AND PATHOGENICITY. SPECIFIC ABLATION OF THIS SUBSET INTERRUPTED THE INFLAMMATORY CASCADE, DESPITE THE UNPERTURBED TISSUE ACCUMULATION OF OTHER TH SUBSETS (E.G., TH1 AND TH17), HIGHLIGHTING THAT GM-CSF EXPRESSION NOT ONLY MARKS PATHOGENIC TH CELLS, BUT THAT THIS SUBSET MEDIATES IMMUNOPATHOLOGY AND TISSUE DESTRUCTION. 2019 2 597 27 BET-BROMODOMAIN AND EZH2 INHIBITOR-TREATED CHRONIC GVHD MICE HAVE BLUNTED GERMINAL CENTERS WITH DISTINCT TRANSCRIPTOMES. DESPITE ADVANCES IN THE FIELD, CHRONIC GRAFT-VERSUS-HOST-DISEASE (CGVHD) REMAINS A LEADING CAUSE OF MORBIDITY AND MORTALITY FOLLOWING ALLOGENIC HEMATOPOIETIC STEM CELL TRANSPLANT. BECAUSE TREATMENT OPTIONS REMAIN LIMITED, WE TESTED EFFICACY OF ANTICANCER, CHROMATIN-MODIFYING ENZYME INHIBITORS IN A CLINICALLY RELEVANT MURINE MODEL OF CGVHD WITH BRONCHIOLITIS OBLITERANS (BO). WE OBSERVED THAT THE NOVEL ENHANCER OF ZESTE HOMOLOG 2 (EZH2) INHIBITOR JQ5 AND THE BET-BROMODOMAIN INHIBITOR JQ1 EACH IMPROVED PULMONARY FUNCTION; IMPAIRED THE GERMINAL CENTER (GC) REACTION, A PREREQUISITE IN CGVHD/BO PATHOGENESIS; AND JQ5 REDUCED EZH2-MEDIATED H3K27ME3 IN DONOR T CELLS. USING CONDITIONAL EZH2 KNOCKOUT DONOR CELLS, WE DEMONSTRATED THAT EZH2 IS OBLIGATORY FOR THE INITIATION OF CGVHD/BO. IN A SCLERODERMATOUS CGVHD MODEL, JQ5 REDUCED THE SEVERITY OF CUTANEOUS LESIONS. TO DETERMINE HOW THE 2 DRUGS COULD LEAD TO THE SAME PHYSIOLOGICAL IMPROVEMENTS WHILE TARGETING UNIQUE EPIGENETIC PROCESSES, WE ANALYZED THE TRANSCRIPTOMES OF SPLENIC GCB CELLS (GCBS) FROM TRANSPLANTED MICE TREATED WITH EITHER DRUG. MULTIPLE INFLAMMATORY AND SIGNALING PATHWAYS ENRICHED IN CGVHD/BO GCBS WERE REDUCED BY EACH DRUG. GCBS FROM JQ5- BUT NOT JQ1-TREATED MICE WERE ENRICHED FOR PROPROLIFERATIVE PATHWAYS ALSO SEEN IN GCBS FROM BONE MARROW-ONLY TRANSPLANTED MICE, LIKELY REFLECTING THEIR UNDERLYING BIOLOGY IN THE UNPERTURBED STATE. IN CONJUNCTION WITH IN VIVO DATA, THESE INSIGHTS LED US TO CONCLUDE THAT EPIGENETIC TARGETING OF THE GC IS A VIABLE CLINICAL APPROACH FOR THE TREATMENT OF CGVHD, AND THAT THE EZH2 INHIBITOR JQ5 AND THE BET-BROMODOMAIN INHIBITOR JQ1 DEMONSTRATED CLINICAL POTENTIAL FOR EZH2I AND BETI IN PATIENTS WITH CGVHD/BO. 2022 3 2914 30 GENE REGULATORY NETWORK UNDERLYING THE IMMORTALIZATION OF EPITHELIAL CELLS. BACKGROUND: TUMORIGENIC TRANSFORMATION OF HUMAN EPITHELIAL CELLS IN VITRO HAS BEEN DESCRIBED EXPERIMENTALLY AS THE POTENTIAL RESULT OF SPONTANEOUS IMMORTALIZATION. THIS PROCESS IS CHARACTERIZED BY A SERIES OF CELL-STATE TRANSITIONS, IN WHICH NORMAL EPITHELIAL CELLS ACQUIRE FIRST A SENESCENT STATE WHICH IS LATER SURPASSED TO ATTAIN A MESENCHYMAL STEM-LIKE PHENOTYPE WITH A POTENTIALLY TUMORIGENIC BEHAVIOR. IN THIS PAPER WE AIM TO PROVIDE A SYSTEM-LEVEL MECHANISTIC EXPLANATION TO THE EMERGENCE OF THESE CELL TYPES, AND TO THE TIME-ORDERED TRANSITION PATTERNS THAT ARE COMMON TO NEOPLASIAS OF EPITHELIAL ORIGIN. TO THIS END, WE FIRST INTEGRATE PUBLISHED FUNCTIONAL AND WELL-CURATED MOLECULAR DATA OF THE COMPONENTS AND INTERACTIONS THAT HAVE BEEN FOUND TO BE INVOLVED IN SUCH CELL STATES AND TRANSITIONS INTO A NETWORK OF 41 MOLECULAR COMPONENTS. WE THEN REDUCE THIS INITIAL NETWORK BY REMOVING SIMPLE MEDIATORS (I.E., LINEAR PATHWAYS), AND FORMALIZE THE RESULTING REGULATORY CORE INTO LOGICAL RULES THAT GOVERN THE DYNAMICS OF EACH OF THE NETWORK COMPONENTS AS A FUNCTION OF THE STATES OF ITS REGULATORS. RESULTS: COMPUTATIONAL DYNAMIC ANALYSIS SHOWS THAT OUR PROPOSED GENE REGULATORY NETWORK MODEL RECOVERS EXACTLY THREE ATTRACTORS, EACH OF THEM DEFINED BY A SPECIFIC GENE EXPRESSION PROFILE THAT CORRESPONDS TO THE EPITHELIAL, SENESCENT, AND MESENCHYMAL STEM-LIKE CELLULAR PHENOTYPES, RESPECTIVELY. WE SHOW THAT ALTHOUGH A MESENCHYMAL STEM-LIKE STATE CAN BE ATTAINED EVEN UNDER UNPERTURBED PHYSIOLOGICAL CONDITIONS, THE LIKELIHOOD OF CONVERGING TO THIS STATE IS INCREASED WHEN PRO-INFLAMMATORY CONDITIONS ARE SIMULATED, PROVIDING A SYSTEMS-LEVEL MECHANISTIC EXPLANATION FOR THE CARCINOGENIC ROLE OF CHRONIC INFLAMMATORY CONDITIONS OBSERVED IN THE CLINIC. WE ALSO FOUND THAT THE REGULATORY CORE YIELDS AN EPIGENETIC LANDSCAPE THAT RESTRICTS TEMPORAL PATTERNS OF PROGRESSION BETWEEN THE STEADY STATES, SUCH THAT RECOVERED PATTERNS RESEMBLE THE TIME-ORDERED TRANSITIONS OBSERVED DURING THE SPONTANEOUS IMMORTALIZATION OF EPITHELIAL CELLS, BOTH IN VIVO AND IN VITRO. CONCLUSION: OUR STUDY STRONGLY SUGGESTS THAT THE IN VITRO TUMORIGENIC TRANSFORMATION OF EPITHELIAL CELLS, WHICH STRONGLY CORRELATES WITH THE PATTERNS OBSERVED DURING THE PATHOLOGICAL PROGRESSION OF EPITHELIAL CARCINOGENESIS IN VIVO, EMERGES FROM UNDERLYING REGULATORY NETWORKS INVOLVED IN EPITHELIAL TRANS-DIFFERENTIATION DURING DEVELOPMENT. 2017 4 2787 25 EZH2-MEDIATED EPIGENETIC MODIFICATION IS REQUIRED FOR ALLOGENEIC T CELL-INDUCED LUPUS DISEASE. BACKGROUND: THE MECHANISMS INVOLVED IN THE PATHOGENESIS OF AUTOIMMUNE DISORDERS, INCLUDING SYSTEMIC LUPUS ERYTHEMATOSUS (SLE), HAVE NOT BEEN FULLY ELUCIDATED. SOME OF THESE MECHANISMS INVOLVE EPIGENETIC REGULATION OF GENE EXPRESSION. THE HISTONE METHYLTRANSFERASE EZH2 CONTRIBUTES TO EPIGENETIC REGULATION OF GENE EXPRESSION, IS HIGHLY EXPRESSED IN GERMINAL CENTER (GC) B CELLS AND FOLLICULAR T HELPER (T(FH)) CELLS, AND MAY BE INVOLVED IN LUPUS PATHOGENESIS. METHODS: THE MURINE BM12 MODEL OF LUPUS-LIKE CHRONIC GRAFT VERSUS HOST DISEASE (CGVHD) WAS INDUCED BY INTRA-PERITONEAL INJECTION OF NEGATIVELY ISOLATED ALLOGENEIC CD4(+) T CELLS. LUPUS-LIKE DISEASE DEVELOPMENT WAS MONITORED BY ELISA DETERMINATION OF SERUM ANTI-DSDNA AND ANTI-CHROMATIN ANTIBODY TITERS. IMMUNE CELL ACTIVATION AND EZH2 EXPRESSION WERE EVALUATED BY FLOW CYTOMETRY AND WESTERN BLOTTING. RESULTS: DECREASED AUTOANTIBODY PRODUCTION AND GC FORMATION ARE OBSERVED WHEN EZH2-DEFICIENT CD4(+) T CELLS ARE USED INSTEAD OF WILD-TYPE (WT) TO INDUCE CGVHD AND WHEN MICE THAT RECEIVE ALLOGENEIC WT DONOR T CELLS TO INDUCE CGVHD ARE TREATED WITH GSK503, AN EZH2-SPECIFIC INHIBITOR. IN THE BM12 CGVHD MODEL, WT DONOR T CELLS ARE NORMALLY FULLY ACTIVATED 1 WEEK AFTER INFUSION INTO AN ALLOGENEIC HOST, EXHIBIT A T(FH) CELL (PD-1(HI)/CXCR5(HI)) PHENOTYPE WITH UPREGULATED EZH2, AND ACTIVATE B CELLS TO FORM GERMINAL CENTERS (GCS). IN CONTRAST, EZH2-DEFICIENT DONOR T CELLS GENERATE FEWER T(FH) CELLS THAT FAIL TO ACTIVATE B CELLS OR PROMOTE GC FORMATION. DESPITE SIMILAR T-INDEPENDENT, LPS-INDUCED B CELL RESPONSES, OVA-IMMUNIZED CD4.EZH2-KO MICE HAD A SKEWED LOW-AFFINITY IGM PHENOTYPE IN COMPARISON TO SIMILARLY TREATED WT MICE. IN ADDITION, EARLY AFTER OVA IMMUNIZATION, MORE CD4(+) T CELLS FROM B6.CD4.EZH2-KO MICE HAD A CD44(LO)/CD62L(LO) PHENOTYPE, WHICH SUGGESTS ARRESTED OR DELAYED ACTIVATION, THAN CD4(+) T CELLS FROM OVALBUMIN-IMMUNIZED B6.WT MICE. CONCLUSION: EZH2 GENE DELETION OR PHARMACOLOGICAL EZH2 INHIBITION SUPPRESSES AUTOANTIBODY PRODUCTION AND GC FORMATION IN BM12 LUPUS-LIKE CGVHD AND DECREASES AFFINITY MATURATION AND ISOTYPE SWITCHING IN RESPONSE TO IMMUNIZATION WITH A T CELL-DEPENDENT ANTIGEN. EZH2 INHIBITION MAY BE USEFUL FOR THE TREATMENT OF LUPUS AND OTHER AUTOIMMUNE DISORDERS. 2020 5 5139 13 POTENTIAL NOVEL BIOMARKERS IN CHRONIC GRAFT-VERSUS-HOST DISEASE. PROGNOSTIC, DIAGNOSTIC OR PREDICTIVE BIOMARKERS ARE URGENTLY NEEDED FOR ASSESSMENT OF CHRONIC GRAFT-VERSUS-HOST DISEASE (CGVHD), A MAJOR RISK FOR PATIENTS UNDERGOING ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION. THE MAIN GOAL OF THIS REVIEW GENERATED WITHIN THE COST ACTION EUROGRAFT "INTEGRATED EUROPEAN NETWORK ON CHRONIC GRAFT VERSUS HOST DISEASE" WAS TO IDENTIFY POTENTIAL NOVEL BIOMARKERS FOR CGVHD BESIDES THE WIDELY ACCEPTED MOLECULAR AND CELLULAR BIOMARKERS. THUS, THE FOCUS WAS ON CELLULAR BIOMARKERS, ALLOANTIBODIES, GLYCOMICS, ENDOTHELIAL DERIVED PARTICLES, EXTRACELLULAR VESICLES, MICROBIOME, EPIGENETIC AND NEUROLOGIC CHANGES IN CGVHD PATIENTS. BOTH HOST-REACTIVE ANTIBODIES IN GENERAL, AND PARTICULARLY ALLOANTIBODIES HAVE BEEN ASSOCIATED WITH CGVHD AND REQUIRE FURTHER CONSIDERATION. GLYCANS ATTACHED TO IGG MODULATE ITS ACTIVITY AND REPRESENT A PROMISING PREDICTIVE AND/OR STRATIFICATION BIOMARKER FOR CGVHD. FURTHERMORE, EPIGENETIC CHANGES SUCH AS MICRORNAS AND DNA METHYLATION REPRESENT POTENTIAL BIOMARKERS FOR MONITORING CGVHD PATIENTS AND NOVEL TARGETS FOR DEVELOPING NEW TREATMENT APPROACHES. FINALLY, THE MICROBIOME LIKELY AFFECTS THE PATHOPHYSIOLOGY OF CGVHD; BACTERIAL STRAINS AS WELL AS MICROBIAL METABOLITES COULD DISPLAY POTENTIAL BIOMARKERS FOR DYSBIOSIS AND RISK FOR THE DEVELOPMENT OF CGVHD. IN SUMMARY, ALTHOUGH THERE ARE NO VALIDATED BIOMARKERS CURRENTLY AVAILABLE FOR CLINICAL USE TO BETTER INFORM ON THE DIAGNOSIS, PROGNOSIS OR PREDICTION OF OUTCOME FOR CGVHD, MANY NOVEL SOURCES OF POTENTIAL MARKERS HAVE SHOWN PROMISE AND WARRANT FURTHER INVESTIGATION USING WELL CHARACTERIZED, MULTI-CENTER PATIENT COHORTS. 2020 6 6117 31 THE EPIGENETIC DRUG TRICHOSTATIN A AMELIORATES EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS VIA T CELL TOLERANCE INDUCTION AND IMPAIRED INFLUX OF T CELLS INTO THE SPINAL CORD. MULTIPLE SCLEROSIS IS A T CELL MEDIATED CHRONIC DEMYELINATING DISEASE OF THE CENTRAL NERVOUS SYSTEM. ALTHOUGH CURRENTLY AVAILABLE THERAPIES REDUCE RELAPSES, THEY DO NOT FACILITATE TOLERIZATION OF MYELIN ANTIGEN-SPECIFIC T LYMPHOCYTES TO ENSURE PROLONGED PROTECTION AGAINST MULTIPLE SCLEROSIS. HERE, WE SHOW THAT TREATMENT OF NOD MICE WITH THE HISTONE DEACETYLASE INHIBITOR, TRICHOSTATIN A AFFORDS ROBUST PROTECTION AGAINST MYELIN PEPTIDE INDUCED EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS, A MOUSE MODEL OF MULTIPLE SCLEROSIS. PROTECTION WAS ACCOMPANIED BY HISTONE HYPERACETYLATION, AND REDUCED INFLAMMATION AND AXONAL DAMAGE IN THE SPINAL CORD. DRUG TREATMENT DIMINISHED THE GENERATION OF CD4(+) MEMORY T CELLS AND INDUCED TOLERANCE IN CD4(+) T CELLS RECOGNIZING THE IMMUNIZING MYELIN PEPTIDE. DURING THE EARLY IMMUNIZATION PERIOD, CD4(+) T CELLS PRODUCING GM-CSF+IFN-GAMMA, GM-CSF+IL-17A, AS WELL AS THOSE EXPRESSING BOTH IL-17A+IFN-GAMMA (DOUBLE-PRODUCERS) WERE DETECTED IN THE SECONDARY LYMPHOID ORGANS FOLLOWED BY THE APPEARANCE OF CELLS PRODUCING IFN-GAMMA AND GM-CSF. ON THE OTHER HAND, IFN-GAMMA PRODUCING TH1 CELLS APPEAR FIRST IN THE SPINAL CORD FOLLOWED BY CELLS PRODUCING IL-17A AND GM-CSF. TREATMENT WITH TRICHOSTATIN A SUBSTANTIALLY REDUCED THE FREQUENCIES OF ALL T CELLS SECRETING VARIOUS LYMPHOKINES BOTH IN THE PERIPHERY AND IN THE SPINAL CORD. THESE DATA INDICATE THAT EPIGENETIC MODIFICATIONS INDUCED BY HISTONE HYPERACETYLATION FACILITATES T CELL TOLERANCE INDUCTION IN THE PERIPHERY LEADING TO REDUCED MIGRATION OF T CELLS TO THE SPINAL CORD AND MITIGATION OF NEURONAL DAMAGE AND IMPROVED CLINICAL OUTCOME. THESE RESULTS SUGGEST THAT EPIGENETIC MODULATION OF THE GENOME MAY SIMILARLY OFFER BENEFITS TO MULTIPLE SCLEROSIS PATIENTS VIA ABROGATING THE FUNCTION OF ENCEPHALITOGENIC T LYMPHOCYTES WITHOUT EXERTING SEVERE SIDE EFFECTS ASSOCIATED WITH CURRENTLY USED DISEASE-MODIFYING THERAPIES. 2017 7 3760 17 INTEGRATED SINGLE CELL ANALYSIS SHOWS CHRONIC ALCOHOL DRINKING DISRUPTS MONOCYTE DIFFERENTIATION IN THE BONE MARROW. CHRONIC HEAVY ALCOHOL DRINKING (CHD) REWIRES MONOCYTES AND MACROPHAGES TOWARD HEIGHTENED INFLAMMATORY STATES WITH COMPROMISED ANTIMICROBIAL DEFENSES THAT PERSIST AFTER 1-MONTH ABSTINENCE. TO DETERMINE WHETHER THESE CHANGES ARE MEDIATED THROUGH ALTERATIONS IN THE BONE MARROW NICHE, WE PROFILED MONOCYTES AND HEMATOPOIETIC STEM CELL PROGENITORS (HSCPS) FROM CHD RHESUS MACAQUES USING A COMBINATION OF FUNCTIONAL ASSAYS AND SINGLE CELL GENOMICS. CHD RESULTED IN TRANSCRIPTIONAL PROFILES CONSISTENT WITH INCREASED ACTIVATION AND INFLAMMATION WITHIN BONE MARROW RESIDENT MONOCYTES AND MACROPHAGES. FURTHERMORE, CHD RESULTED IN TRANSCRIPTIONAL SIGNATURES ASSOCIATED WITH INCREASED OXIDATIVE AND CELLULAR STRESS IN HSCP. DIFFERENTIATION OF HSCP IN VITRO REVEALED SKEWING TOWARD MONOCYTES EXPRESSING "NEUTROPHIL-LIKE" MARKERS WITH GREATER INFLAMMATORY RESPONSES TO BACTERIAL AGONISTS. FURTHER ANALYSES OF HSCPS SHOWED BROAD EPIGENETIC CHANGES THAT WERE IN LINE WITH EXACERBATED INFLAMMATORY RESPONSES WITHIN MONOCYTES AND THEIR PROGENITORS. IN SUMMARY, CHD ALTERS HSCPS IN THE BONE MARROW LEADING TO THE PRODUCTION OF MONOCYTES POISED TO GENERATE DYSREGULATED HYPER-INFLAMMATORY RESPONSES. 2023 8 1906 25 ENHANCER OF ZESTE HOMOLOG 2-CATALYSED H3K27 TRIMETHYLATION PLAYS A KEY ROLE IN ACUTE-ON-CHRONIC LIVER FAILURE VIA TNF-MEDIATED PATHWAY. ACUTE-ON-CHRONIC LIVER FAILURE IS MAINLY DUE TO HOST IMMUNITY SELF-DESTRUCTION. THE HISTONE H3 LYSINE 27 (H3K27) TRIMETHYLATING ENZYME, ENHANCER OF ZESTE HOMOLOG 2 (EZH2) MEDIATES EPIGENETIC SILENCING OF GENE EXPRESSION AND REGULATES IMMUNITY, ALSO INVOLVES PATHOGENESIS OF SEVERAL LIVER DISEASES. THE CURRENT STUDY WAS TO DETERMINE THE ROLE OF METHYLTRANSFERASE EZH2 AND ITS CATALYSED H3K27 TRIMETHYLATION (H3K27ME3) IN LIVER FAILURE, AND TO FURTHER INVESTIGATE THE POTENTIAL TARGET FOR LIVER FAILURE TREATMENT. EZH2 AND ITS CATALYSED H3K27ME3 WERE DETERMINED IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMC) FROM LIVER FAILURE PATIENTS AND KUPFFER CELLS FROM EXPERIMENTAL MICE. FURTHERMORE, GSK126 (AN INHIBITOR FOR EZH2 TRIMETHYLATION FUNCTION) WAS APPLIED IN LIVER FAILURE MICE IN VIVO, AND LIPOPOLYSACCHARIDE-STIMULATED MONONUCLEAR CELLS IN VITRO. EZH2 AND H3K27ME3 WERE SIGNIFICANTLY UPREGULATED IN HUMAN PBMC FROM LIVER FAILURE PATIENTS OR MURINE KUPFFER CELLS FROM THE LIVER FAILURE ANIMALS, RESPECTIVELY. GSK126 AMELIORATED DISEASE SEVERITY IN LIVER FAILURE MICE, WHICH MAYBE ATTRIBUTE TO DOWN-REGULATE CIRCULATING AND HEPATIC PROINFLAMMATORY CYTOKINES, ESPECIALLY TNF VIA REDUCING H3K27ME3. IN-DEPTH CHROMATIN IMMUNOPRECIPITATION ANALYSIS UNRAVELLED THAT DECREASED ENRICHMENT OF H3K27ME3 ON TNF PROMOTOR, RESULTING IN TNF ELEVATION IN KUPFFER CELLS FROM LIVER FAILURE MICE. NUCLEAR FACTOR KAPPA B (NF-KAPPAB) AND PROTEIN KINASE B (AKT) SIGNALLING PATHWAYS WERE ACTIVATED UPON LIPOPOLYSACCHARIDE STIMULATION, BUT ATTENUATED BY USING GSK126, ACCOMPANIED WITH DECREASED TNF IN VITRO. IN CONCLUSION, EZH2 AND H3K27ME3 CONTRIBUTED TO THE PATHOGENESIS OF LIVER FAILURE VIA TRIGGERING TNF AND OTHER INDISPENSABLE PROINFLAMMATORY CYTOKINES. EZH2 WAS TO MODIFY H3K27ME3 ENRICHMENT, AS WELL AS, ACTIVATION OF THE DOWNSTREAM NF-KAPPAB AND AKT SIGNALLING PATHWAYS. 2018 9 1061 24 CLINICAL REMISSION OF SIGHT-THREATENING NON-INFECTIOUS UVEITIS IS CHARACTERIZED BY AN UPREGULATION OF PERIPHERAL T-REGULATORY CELL POLARIZED TOWARDS T-BET AND TIGIT. BACKGROUND: NON-INFECTIOUS UVEITIS CAN CAUSE CHRONIC RELAPSING AND REMITTING OCULAR INFLAMMATION, WHICH MAY REQUIRE HIGH DOSE SYSTEMIC IMMUNOSUPPRESSION TO PREVENT SEVERE SIGHT LOSS. IT HAS BEEN CLASSICALLY DESCRIBED AS AN AUTOIMMUNE DISEASE, MEDIATED BY PRO-INFLAMMATORY TH1 AND TH17 T-CELL SUBSETS. STUDIES SUGGEST THAT NATURAL IMMUNOSUPPRESSIVE CD4(+)CD25(+)FOXP3(+) T-REGULATORY CELLS (TREGS) ARE INVOLVED IN RESOLUTION OF INFLAMMATION AND MAY BE INVOLVED IN THE MAINTENANCE OF CLINICAL REMISSION. OBJECTIVE: TO INVESTIGATE WHETHER THERE IS A PERIPHERAL BLOOD IMMUNOREGULATORY PHENOTYPE ASSOCIATED WITH CLINICAL REMISSION OF SIGHT-THREATENING NON-INFECTIOUS UVEITIS BY COMPARING PERIPHERAL BLOOD LEVELS OF TREG, TH1, AND TH17, AND ASSOCIATED DNA METHYLATION AND CYTOKINE LEVELS IN PATIENTS WITH ACTIVE UVEITIC DISEASE, CONTROL SUBJECTS AND PATIENTS (WITH PREVIOUSLY ACTIVE DISEASE) IN CLINICAL REMISSION INDUCED BY IMMUNOSUPPRESSIVE DRUGS. METHODS: ISOLATED PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMC) FROM PERIPHERAL BLOOD SAMPLES FROM PROSPECTIVELY RECRUITED SUBJECTS WERE ANALYZED BY FLOW CYTOMETRY FOR CD3, CD4, FOXP3, TIGIT, T-BET, AND RELATED ORPHAN RECEPTOR GAMMAT. EPIGENETIC DNA METHYLATION LEVELS OF FOXP3 TREG-SPECIFIC DEMETHYLATED REGION (TSDR), FOXP3 PROMOTER, TBX21, RORC2, AND TIGIT LOCI WERE DETERMINED IN CRYOPRESERVED PBMC USING A NEXT-GENERATION SEQUENCING APPROACH. RELATED CYTOKINES WERE MEASURED IN BLOOD SERA. FUNCTIONAL SUPPRESSIVE CAPACITY OF TREG WAS ASSESSED USING T-CELL PROLIFERATION ASSAYS. RESULTS: FIFTY PATIENTS WITH UVEITIS (INTERMEDIATE, POSTERIOR, AND PANUVEITIS) AND 10 CONTROL SUBJECTS WERE RECRUITED. THE FREQUENCY OF CD4(+)CD25(+)FOXP3(+) TREG, TIGIT(+) TREG, AND T-BET(+) TREG AND THE RATIO OF TREG TO TH1 WERE SIGNIFICANTLY HIGHER IN REMISSION PATIENTS COMPARED WITH PATIENTS WITH ACTIVE UVEITIC DISEASE; AND TIGIT(+) TREGS WERE A SIGNIFICANT PREDICTOR OF CLINICAL REMISSION. TREG FROM PATIENTS IN CLINICAL REMISSION DEMONSTRATED A HIGH LEVEL OF IN VITRO SUPPRESSIVE FUNCTION COMPARED WITH TREG FROM CONTROL SUBJECTS AND FROM PATIENTS WITH UNTREATED ACTIVE DISEASE. PBMC FROM PATIENTS IN CLINICAL REMISSION HAD SIGNIFICANTLY LOWER METHYLATION LEVELS AT THE FOXP3 TSDR, FOXP3 PROMOTER, AND TIGIT LOCI AND HIGHER LEVELS AT RORC LOCI THAN THOSE WITH ACTIVE DISEASE. CLINICAL REMISSION WAS ALSO ASSOCIATED WITH SIGNIFICANTLY HIGHER SERUM LEVELS OF TRANSFORMING GROWTH FACTOR BETA AND IL-10, WHICH POSITIVELY CORRELATED WITH TREG LEVELS, AND LOWER SERUM LEVELS OF IFNGAMMA, IL-17A, AND IL-22 COMPARED WITH PATIENTS WITH ACTIVE DISEASE. CONCLUSION: CLINICAL REMISSION OF SIGHT-THREATENING NON-INFECTIOUS UVEITIS HAS AN IMMUNOREGULATORY PHENOTYPE CHARACTERIZED BY UPREGULATION OF PERIPHERAL TREG, POLARIZED TOWARD T-BET AND TIGIT. THESE FINDINGS MAY ASSIST WITH INDIVIDUALIZED THERAPY OF UVEITIS, BY INFORMING WHETHER DRUG THERAPY HAS INDUCED PHENOTYPICALLY STABLE TREG ASSOCIATED WITH LONG-TERM CLINICAL REMISSION. 2018 10 2016 18 EPIGENETIC BIOMARKERS IN PROGRESSION FROM NON-DYSPLASTIC BARRETT'S OESOPHAGUS TO OESOPHAGEAL ADENOCARCINOMA: A SYSTEMATIC REVIEW PROTOCOL. INTRODUCTION: BARRETT'S OESOPHAGUS (BO), A METAPLASTIC CONDITION AFFECTING THE LOWER OESOPHAGUS DUE TO LONG-STANDING GASTRO-OESOPHAGEAL REFLUX AND CHRONIC INFLAMMATION, IS A PRECURSOR LESION FOR OESOPHAGEAL ADENOCARCINOMA (OADC). THERE IS NO CLINICAL TEST TO PREDICT WHICH PATIENTS WITH BO WILL PROGRESS TO OADC. THE BRITISH SOCIETY OF GASTROENTEROLOGY RECOMMENDS ENDOSCOPIC SURVEILLANCE OF PATIENTS WITH BO. EPIGENETIC CHANGES HAVE BEEN WELL CHARACTERISED IN THE NEOPLASTIC PROGRESSION OF ULCERATIVE COLITIS TO COLONIC CARCINOMA, ANOTHER GASTROINTESTINAL CANCER ASSOCIATED WITH CHRONIC INFLAMMATION. THIS SYSTEMATIC REVIEW PROTOCOL AIMS TO IDENTIFY AND EVALUATE STUDIES WHICH EXAMINE EPIGENETIC BIOMARKERS IN BO AND THEIR ASSOCIATION WITH PROGRESSION TO OADC. METHODS AND ANALYSIS: ALL PROSPECTIVE AND RETROSPECTIVE PRIMARY STUDIES, AND EXISTING SYSTEMATIC REVIEWS INVESTIGATING EPIGENETIC MARKERS INCLUDING DNA METHYLATION, HISTONE MODIFICATION, CHROMATIN REMODELLING, MICRO AND NON-CODING RNAS OF ALL TYPES WILL BE ELIGIBLE FOR INCLUSION. ELIGIBLE PATIENTS ARE THOSE OVER THE AGE OF 18 WITH BO, BO WITH DYSPLASIA, OADC OR UNSPECIFIED OESOPHAGEAL CANCER. A COMPREHENSIVE SEARCH OF BIBLIOGRAPHIC DATABASES USING COMBINATIONS OF TEXT AND INDEX WORDS RELATING TO THE POPULATION, PROGNOSTIC MARKERS AND OUTCOME WILL BE UNDERTAKEN WITH NO LANGUAGE RESTRICTIONS. RESULTS WILL BE SCREENED BY 2 INDEPENDENT REVIEWERS AND DATA EXTRACTED USING A STANDARDISED PROFORMA. THE QUALITY AND RISK OF BIAS OF INDIVIDUAL STUDIES WILL BE ASSESSED USING THE QUALITY IN PROGNOSTIC STUDIES (QUIPS) TOOL. A NARRATIVE SYNTHESIS OF ALL EVIDENCE WILL BE PERFORMED WITH KEY FINDINGS TABULATED. META-ANALYSIS WILL BE CONSIDERED WHERE STUDIES AND REPORTED OUTCOMES ARE CONSIDERED SUFFICIENTLY HOMOGENEOUS, BOTH CLINICALLY AND METHODOLOGICALLY. FINDINGS WILL BE INTERPRETED IN THE CONTEXT OF THE QUALITY OF INCLUDED STUDIES. THE SYSTEMATIC REVIEW WILL BE REPORTED ACCORDING TO PRISMA GUIDELINES. ETHICS AND DISSEMINATION: THIS IS A SYSTEMATIC REVIEW OF COMPLETED STUDIES AND NO ETHICAL APPROVAL IS REQUIRED. FINDINGS FROM THE FULL SYSTEMATIC REVIEW WILL BE SUBMITTED FOR PUBLICATION AND PRESENTATION AT NATIONAL AND INTERNATIONAL CONFERENCES WHICH WILL INFORM FUTURE RESEARCH ON RISK STRATIFICATION IN PATIENTS WITH BO. REVIEW REGISTRATION NUMBER: CRD42016038654. 2016 11 1472 32 DISTINCT MECHANISMS REGULATE IL1B GENE TRANSCRIPTION IN LYMPHOID CD4 T CELLS AND MONOCYTES. INTERLEUKIN 1BETA IS A PRO-INFLAMMATORY CYTOKINE IMPORTANT FOR BOTH NORMAL IMMUNE RESPONSES AND CHRONIC INFLAMMATORY DISEASES. THE REGULATION OF THE 31 KDA PROIL-1BETA PRECURSOR CODED BY THE IL1B GENE HAS BEEN EXTENSIVELY STUDIED IN MYELOID CELLS, BUT NOT IN LYMPHOID-DERIVED CD4 T CELLS. SURPRISINGLY, WE FOUND THAT SOME CD4 T CELL SUBSETS EXPRESS HIGHER LEVELS OF PROIL-1BETA THAN UNSTIMULATED MONOCYTES, DESPITE RELATIVELY LOW IL1B MRNA LEVELS. WE OBSERVED A SIGNIFICANT INCREASE IN IL1B TRANSCRIPTION AND TRANSLATION IN CD4 T CELLS UPON EX VIVO CD3/CD28 ACTIVATION, AND A SIMILAR ELEVATION IN THE CCR5+ EFFECTOR MEMORY POPULATION COMPARED TO CCR5- T CELLS IN VIVO. THE RAPID AND VIGOROUS INCREASE IN IL1B GENE TRANSCRIPTION FOR STIMULATED MONOCYTES HAS PREVIOUSLY BEEN ASSOCIATED WITH THE PRESENCE OF SPI-1/PU.1 (SPI1), A MYELOID-LINEAGE TRANSCRIPTION FACTOR, PRE-BOUND TO THE PROMOTER. IN THE CASE OF CD4 T CELLS, THIS INCREASE OCCURRED DESPITE THE LACK OF DETECTABLE SPI1 AT THE IL1B PROMOTER. ADDITIONALLY, WE FOUND ALTERED EPIGENETIC REGULATION OF THE IL1B LOCUS IN CD3/CD28-ACTIVATED CD4 T CELLS. UNLIKE MONOCYTES, ACTIVATED CD4 T CELLS POSSESS BIVALENT H3K4ME3+/H3K27ME3+ NUCLEOSOME MARKS AT THE IL1B PROMOTER, REFLECTING LOW TRANSCRIPTIONAL ACTIVITY. THESE RESULTS SUPPORT A MODEL IN WHICH THE IL1B GENE IN CD4 T CELLS IS TRANSCRIBED FROM A LOW-ACTIVITY BIVALENT PROMOTER INDEPENDENT OF SPI1. ACCUMULATED CYTOPLASMIC PROIL-1BETA MAY ULTIMATELY BE CLEAVED TO MATURE 17 KDA BIOACTIVE IL-1BETA, REGULATING T CELL POLARIZATION AND PATHOGENIC CHRONIC INFLAMMATION. 2018 12 2063 25 EPIGENETIC CONTROL OF IL-23 EXPRESSION IN KERATINOCYTES IS IMPORTANT FOR CHRONIC SKIN INFLAMMATION. THE CHRONIC SKIN INFLAMMATION PSORIASIS IS CRUCIALLY DEPENDENT ON THE IL-23/IL-17 CYTOKINE AXIS. ALTHOUGH IL-23 IS EXPRESSED BY PSORIATIC KERATINOCYTES AND IMMUNE CELLS, ONLY THE IMMUNE CELL-DERIVED IL-23 IS BELIEVED TO BE DISEASE RELEVANT. HERE WE USE A GENETIC MOUSE MODEL TO SHOW THAT KERATINOCYTE-PRODUCED IL-23 IS SUFFICIENT TO CAUSE A CHRONIC SKIN INFLAMMATION WITH AN IL-17 PROFILE. FURTHERMORE, WE REVEAL A CELL-AUTONOMOUS NUCLEAR FUNCTION FOR THE ACTIN POLYMERIZING MOLECULE N-WASP, WHICH CONTROLS IL-23 EXPRESSION IN KERATINOCYTES BY REGULATING THE DEGRADATION OF THE HISTONE METHYLTRANSFERASES G9A AND GLP, AND H3K9 DIMETHYLATION OF THE IL-23 PROMOTER. THIS MECHANISM MEDIATES THE INDUCTION OF IL-23 BY TNF, A KNOWN INDUCER OF IL-23 IN PSORIASIS. FINALLY, IN KERATINOCYTES OF PSORIATIC LESIONS A DECREASE IN H3K9 DIMETHYLATION CORRELATES WITH INCREASED IL-23 EXPRESSION, SUGGESTING RELEVANCE FOR DISEASE. TAKEN TOGETHER, OUR DATA DESCRIBE A MOLECULAR PATHWAY WHERE EPIGENETIC REGULATION OF KERATINOCYTES CAN CONTRIBUTE TO CHRONIC SKIN INFLAMMATION. 2018 13 1905 25 ENHANCER OF ZESTE HOMOLOG 2 CONTRIBUTES TO APOPTOSIS BY INACTIVATING JANUS KINASE 2/ SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION SIGNALING IN INFLAMMATORY BOWEL DISEASE. BACKGROUND: INFLAMMATORY BOWEL DISEASE (IBD) IS A PREVALENT WORLDWIDE HEALTH PROBLEM FEATURED BY RELAPSING, CHRONIC GASTROINTESTINAL INFLAMMATION. ENHANCER OF ZESTE HOMOLOG 2 (EZH2) IS A CRITICAL EPIGENETIC REGULATOR IN DIFFERENT PATHOLOGICAL MODELS, SUCH AS CANCER AND INFLAMMATION. HOWEVER, THE ROLE OF EZH2 IN THE IBD DEVELOPMENT IS STILL OBSCURE. AIM: TO EXPLORE THE EFFECT OF EZH2 ON IBD PROGRESSION AND THE UNDERLYING MECHANISM. METHODS: THE IBD MOUSE MODEL WAS CONDUCTED BY ADDING DEXTRAN SODIUM SULFATE (DSS), AND THE EFFECT OF EZH2 ON DSS-INDUCED COLITIS WAS ASSESSED IN THE MODEL. THE FUNCTION OF EZH2 IN REGULATING APOPTOSIS AND PERMEABILITY WAS EVALUATED BY ANNEXIN V-FITC APOPTOSIS DETECTION KIT, TRANSEPITHELIAL ELECTRICAL RESISTANCE ANALYSIS, AND WESTERN BLOT ANALYSIS OF RELATED MARKERS, INCLUDING ZONA OCCLUDENS 1, CLAUDIN-5, AND OCCLUDIN, IN NCM460 AND FETAL HUMAN COLON (FHC) CELLS. THE MECHANICAL INVESTIGATION WAS PERFORMED BY QUANTITATIVE REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION, WESTERN BLOT ANALYSIS, AND CHROMATIN IMMUNOPRECIPITATION ASSAYS. RESULTS: THE COLON LENGTH WAS INHIBITED IN THE DSS-TREATED MICE AND WAS ENHANCED BY THE EZH2 DEPLETION IN THE SYSTEM. DSS TREATMENT CAUSED A DECREASED HISTOLOGICAL SCORE IN THE MICE, WHICH WAS REVERSED BY EZH2 DEPLETION. THE INFLAMMATORY CYTOKINES, SUCH AS TUMOR NECROSIS FACTOR-ALPHA, INTERLEUKIN-6, AND INTERLEUKIN-1BETA, WERE INDUCED IN THE DSS-TREATED MICE, IN WHICH THE DEPLETION OF EZH2 COULD REVERSE THIS EFFECT. MOREOVER, THE TUMOR NECROSIS FACTOR-ALPHA TREATMENT INDUCED THE APOPTOSIS OF NCM460 AND FHC CELLS, IN WHICH EZH2 DEPLETION COULD REVERSE THIS EFFECT IN THE CELLS. MOREOVER, THE DEPLETION OF EZH2 ATTENUATED PERMEABILITY OF COLONIC EPITHELIAL CELLS. MECHANICALLY, THE DEPLETION OF EZH2 OR EZH2 INHIBITOR GSK343 WAS ABLE TO ENHANCE THE EXPRESSION AND THE PHOSPHORYLATION OF JANUS KINASE 2 (JK2) AND SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION IN THE NCM460 AND FHC CELLS. SPECIFICALLY, EZH2 INACTIVATED JAK2 EXPRESSION BY REGULATING HISTONE H3K27ME3. JAK2 INHIBITOR TG101348 WAS ABLE TO REVERSE EZH2 KNOCKDOWN-MEDIATED COLONIC EPITHELIAL CELL PERMEABILITY AND APOPTOSIS. CONCLUSION: THUS, WE CONCLUDED THAT EZH2 CONTRIBUTED TO APOPTOSIS AND INFLAMMATORY RESPONSE BY INACTIVATING JAK2/ SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION SIGNALING IN IBD. EZH2 MAY BE APPLIED AS A POTENTIAL TARGET FOR IBD THERAPY. 2021 14 2067 26 EPIGENETIC CONTROL OF MACROPHAGE SHAPE TRANSITION TOWARDS AN ATYPICAL ELONGATED PHENOTYPE BY HISTONE DEACETYLASE ACTIVITY. INFLAMMATORY CHRONIC PATHOLOGIES ARE COMPLEX PROCESSES CHARACTERIZED BY AN IMBALANCE BETWEEN THE RESOLUTION OF THE INFLAMMATORY PHASE AND THE ESTABLISHMENT OF TISSUE REPAIR. THE MAIN PLAYERS IN THESE INFLAMMATORY PATHOLOGIES ARE BONE MARROW DERIVED MONOCYTES (BMDMS). HOWEVER, HOW MONOCYTE DIFFERENTIATION IS MODULATED TO GIVE RISE TO SPECIFIC MACROPHAGE SUBPOPULATIONS (M1 OR M2) THAT MAY EITHER MAINTAIN THE CHRONIC INFLAMMATORY PROCESS OR LEAD TO WOUND HEALING IS STILL UNCLEAR. CONSIDERING THAT INHIBITORS OF HISTONE DEACETYLASE (HDAC) HAVE AN ANTI-INFLAMMATORY ACTIVITY, WE ASKED WHETHER THIS ENZYME WOULD PLAY A ROLE ON MONOCYTE DIFFERENTIATION INTO M1 OR M2 PHENOTYPE AND IN THE CELL SHAPE TRANSITION THAT FOLLOWS. WE THEN INDUCED MURINE BONE MARROW PROGENITORS INTO MONOCYTE/MACROPHAGE DIFFERENTIATION PATHWAY USING MEDIA CONTAINING GM-CSF AND THE HDAC BLOCKER, TRICHOSTATIN A (TSA). WE FOUND THAT THE PHARMACOLOGICAL INHIBITION OF HDAC ACTIVITY LED TO A SHAPE TRANSITION FROM THE TYPICAL MACROPHAGE PANCAKE-LIKE SHAPE INTO AN ELONGATED MORPHOLOGY, WHICH WAS CORRELATED TO A MIXED M1/M2 PROFILE OF CYTOKINE AND CHEMOKINE SECRETION. OUR RESULTS PRESENT, FOR THE FIRST TIME, THAT HDAC ACTIVITY ACTS AS A REGULATOR OF MACROPHAGE DIFFERENTIATION IN THE ABSENCE OF LYMPHOCYTE STIMULI. WE PROPOSE THAT HDAC ACTIVITY DOWN REGULATES MACROPHAGE PLASTICITY FAVORING THE PRO-INFLAMMATORY PHENOTYPE. 2015 15 2784 20 EZH2 PROMOTES EXTRACELLULAR MATRIX DEGRADATION VIA NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN PULPITIS. PULPITIS IS A COMPLICATED CHRONIC INFLAMMATORY PROCESS WHICH CAN BE IN A DYNAMIC BALANCE BETWEEN DAMAGE AND REPAIR. THE EXTRACELLULAR MATRIX PLAYS AN IMPORTANT REGULATORY ROLE IN WOUND HEALING AND TISSUE REPAIR. THE AIM OF THIS STUDY WAS TO EXPLORE THE ROLE OF THE EPIGENETIC MARK, ENHANCER OF ZESTE HOMOLOG 2 (EZH2) ON THE DEGRADATION OF EXTRACELLULAR MATRIX DURING PULPITIS. QUANTITATIVE POLYMERASE CHAIN REACTION WAS USED TO ASSESS THE EXPRESSION OF MATRIX METALLOPROTEINASES (MMPS) AND TYPE I COLLAGEN IN HUMAN DENTAL PULP CELLS (HDPCS) UPON EZH2 AND EI1 (EZH2 INHIBITOR) STIMULATION. THE MECHANISM OF EZH2 AFFECTING EXTRACELLULAR MATRIX WAS EXPLORED THROUGH QUANTITATIVE POLYMERASE CHAIN REACTION AND WESTERN BLOT. A RAT MODEL OF DENTAL PULP INFLAMMATION WAS ESTABLISHED, AND THE EXPRESSION OF TYPE I COLLAGEN IN DENTAL PULP UNDER EZH2 STIMULATION WAS DETECTED BY IMMUNOHISTOCHEMICAL STAINING. EZH2 UPREGULATED THE EXPRESSION OF MMP-1, MMP-3, MMP-8, AND MMP-10 AND DECREASED THE PRODUCTION OF TYPE I COLLAGEN IN HDPCS, WHILE EI1 HAD THE OPPOSITE EFFECT. EZH2 ACTIVATED THE NUCLEAR FACTOR-KAPPA B (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN HDPCS, THE INHIBITION OF WHICH REVERSED THE INDUCTION OF MMPS AND THE SUPPRESSION OF TYPE I COLLAGEN. EZH2 CAN DOWNREGULATE THE TYPE I COLLAGEN LEVELS IN AN EXPERIMENTAL MODEL OF DENTAL PULPITIS IN RATS. EZH2 PROMOTES EXTRACELLULAR MATRIX DEGRADATION VIA NUCLEAR FACTOR-KAPPAB (NF-KAPPAB) AND P38 SIGNALING PATHWAYS IN PULPITIS. EZH2 CAN DECREASE THE TYPE I COLLAGEN LEVELS IN VIVO AND IN VITRO. 2021 16 1469 18 DISTINCT EVOLUTIONARY PATHS IN CHRONIC LYMPHOCYTIC LEUKEMIA DURING RESISTANCE TO THE GRAFT-VERSUS-LEUKEMIA EFFECT. LEUKEMIC RELAPSE REMAINS A MAJOR BARRIER TO SUCCESSFUL ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION (ALLO-HSCT) FOR AGGRESSIVE HEMATOLOGIC MALIGNANCIES. THE BASIS FOR RELAPSE OF ADVANCED LYMPHOID MALIGNANCIES REMAINS INCOMPLETELY UNDERSTOOD AND MAY INVOLVE ESCAPE FROM THE GRAFT-VERSUS-LEUKEMIA (GVL) EFFECT. WE HYPOTHESIZED THAT FOR PATIENTS WITH CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) TREATED WITH ALLO-HSCT, LEUKEMIC CELL-INTRINSIC FEATURES INFLUENCE TRANSPLANT OUTCOMES BY DIRECTING THE EVOLUTIONARY TRAJECTORIES OF CLL CELLS. INTEGRATED GENETIC, TRANSCRIPTOMIC, AND EPIGENETIC ANALYSES OF CLL CELLS FROM 10 PATIENTS REVEALED THAT THE CLINICAL KINETICS OF POST-HSCT RELAPSE ARE SHAPED BY DISTINCT MOLECULAR DYNAMICS. EARLY RELAPSES AFTER ALLO-HSCT EXHIBITED NOTABLE GENETIC STABILITY; SINGLE CLL CELL TRANSCRIPTIONAL ANALYSIS DEMONSTRATED A CELLULAR HETEROGENEITY THAT WAS STATIC OVER TIME. IN CONTRAST, CLL CELLS RELAPSING LATE AFTER ALLO-HSCT DISPLAYED NOTABLE GENETIC EVOLUTION AND EVIDENCE OF NEOANTIGEN DEPLETION, CONSISTENT WITH MARKED SINGLE-CELL TRANSCRIPTIONAL SHIFTS THAT WERE UNIQUE TO EACH PATIENT. WE OBSERVED A GREATER RATE OF EPIGENETIC CHANGE FOR LATE RELAPSES NOT SEEN IN EARLY RELAPSES OR RELAPSES AFTER CHEMOTHERAPY ALONE, SUGGESTING THAT THE SELECTION PRESSURES OF THE GVL BOTTLENECK ARE UNLIKE THOSE IMPOSED BY CHEMOTHERAPY. NO SELECTIVE ADVANTAGE FOR HUMAN LEUKOCYTE ANTIGEN (HLA) LOSS WAS OBSERVED, EVEN WHEN PRESENT IN PRETRANSPLANT SUBPOPULATIONS. GAIN OF STEM CELL MODULES WAS A COMMON SIGNATURE ASSOCIATED WITH LEUKEMIA RELAPSE REGARDLESS OF POSTTRANSPLANT RELAPSE KINETICS. THESE DATA ELUCIDATE THE BIOLOGICAL PATHWAYS THAT UNDERLIE GVL RESISTANCE AND POSTTRANSPLANT RELAPSE. 2020 17 5602 31 RORGAMMAT(+) HEMATOPOIETIC CELLS ARE NECESSARY FOR TUMOR CELL PROLIFERATION DURING COLITIS-ASSOCIATED TUMORIGENESIS IN MICE. COLORECTAL CANCER (CRC) IS ONE OF THE MOST COMMON TUMOR ENTITIES. IN PATIENTS WITH INFLAMMATORY BOWEL DISEASES, THE DEVELOPMENT OF COLITIS-ASSOCIATED COLON CANCER IS CONSIDERED A DANGEROUS LONG-TERM COMPLICATION. IL-17A AND THE TRANSCRIPTION FACTOR RETINOIC ACID RECEPTOR-RELATED ORPHAN RECEPTOR GAMMAT (RORGAMMAT) PLAY FUNDAMENTAL ROLES IN THE PATHOGENESIS OF INFLAMMATORY BOWEL DISEASES; IN HUMAN STUDIES, WE DETECTED A DENSE INFILTRATION OF RORGAMMAT-DEPENDENT CD4(+) IL17A(+) T HELPER (TH)17 CELLS IN SPECIMENS OF CRC, ULCERATIVE COLITIS, AND ULCERATIVE COLITIS-ASSOCIATED COLORECTAL CANCER. HOWEVER, THE MECHANISTIC ROLE OF RORGAMMAT(+) HEMATOPOIETIC CELLS IN COLITIS-ASSOCIATED TUMORIGENESIS REMAINS UNCLEAR. TO INVESTIGATE COLITIS-ASSOCIATED COLON TUMORIGENESIS, WE CONDUCTED STUDIES IN THE AOM+DSS MOUSE MODEL THAT REVEALED THE IMPORTANCE OF RORGAMMAT FOR COLON TUMOR PROGRESSION. IN THE ABSENCE OF RORGAMMAT-DEPENDENT TH17 LYMPHOCYTES, MICE SHOWED SIGNS OF INTENSE CHRONIC COLITIS, BUT DEVELOPED SIGNIFICANTLY FEWER MACROSCOPIC TUMOR NODULES. THE REDUCTION OF TUMOR DEVELOPMENT IN RORGAMMAT(-/-) MICE WAS NOT DUE TO REDUCED COLON TUMOR INITIATION. HOWEVER, THE PROLIFERATION RATE OF TUMOR CELLS WAS REDUCED IN THE ABSENCE OF RORGAMMAT-DEPENDENT TH17 CELLS AND TUMOR CELLS SHOWED PRONOUNCED SIGNS OF SENESCENCE-ASSOCIATED EPIGENETIC AND LYSOSOMAL CHANGES. THESE RESULTS INDICATE AN IMPORTANT ROLE FOR THE IMMUNOLOGICAL MILIEU IN COLITIS-ASSOCIATED CANCER, WHICH IS SHAPED IN-PART BY RORGAMMAT-DEPENDENT TH17 LYMPHOCYTES THAT SUPPORT CRC GROWTH. 2015 18 3759 20 INTEGRATED SINGLE CELL ANALYSIS SHOWS CHRONIC ALCOHOL DRINKING DISRUPTS MONOCYTE DIFFERENTIATION IN THE BONE MARROW NICHE. CHRONIC ALCOHOL DRINKING REWIRES CIRCULATING MONOCYTES AND TISSUE-RESIDENT MACROPHAGES TOWARDS HEIGHTENED INFLAMMATORY STATES WITH COMPROMISED ANTI-MICROBIAL DEFENSES. AS THESE EFFECTS REMAIN CONSISTENT IN SHORT-LIVED MONOCYTES AFTER A 1-MONTH ABSTINENCE PERIOD IT IS UNCLEAR WHETHER THESE CHANGES ARE RESTRICTED TO THE PERIPHERY OR MEDIATED THROUGH ALTERATIONS IN THE PROGENITOR NICHE. TO TEST THIS HYPOTHESIS, WE PROFILED MONOCYTES/MACROPHAGES AND HEMATOPOIETIC STEM CELL PROGENITORS (HSCP) OF THE BONE MARROW COMPARTMENT FROM RHESUS MACAQUES AFTER 12 MONTHS OF ETHANOL CONSUMPTION USING A COMBINATION OF FUNCTIONAL ASSAYS AND SINGLE CELL GENOMICS. BONE MARROW-RESIDENT MONOCYTES/MACROPHAGES FROM ETHANOL-CONSUMING ANIMALS EXHIBITED HEIGHTENED INFLAMMATION. DIFFERENTIATION OF HSCP IN VITRO REVEALED SKEWING TOWARDS MONOCYTES EXPRESSING NEUTROPHIL-LIKE MARKERS WITH HEIGHTENED INFLAMMATORY RESPONSES TO BACTERIAL AGONISTS. SINGLE CELL TRANSCRIPTIONAL ANALYSIS OF HSCPS SHOWED REDUCED PROLIFERATION BUT INCREASED INFLAMMATORY MARKERS IN MATURE MYELOID PROGENITORS. WE OBSERVED TRANSCRIPTIONAL SIGNATURES ASSOCIATED WITH INCREASED OXIDATIVE AND CELLULAR STRESS AS WELL AS OXIDATIVE PHOSPHORYLATION IN IMMATURE AND MATURE MYELOID PROGENITORS. SINGLE CELL ANALYSIS OF THE CHROMATIN LANDSCAPE SHOWED ALTERED DRIVERS OF DIFFERENTIATION IN MONOCYTES AND PROGENITORS. COLLECTIVELY, THESE DATA INDICATE THAT CHRONIC ETHANOL DRINKING RESULTS IN REMODELING OF THE TRANSCRIPTIONAL AND EPIGENETIC LANDSCAPES OF THE BONE MARROW COMPARTMENT LEADING TO ALTERED FUNCTIONS IN THE PERIPHERY. 2023 19 6540 34 TRANSCRIPTIONAL, EPIGENETIC, AND FUNCTIONAL REPROGRAMMING OF MONOCYTES FROM NON-HUMAN PRIMATES FOLLOWING CHRONIC ALCOHOL DRINKING. CHRONIC HEAVY DRINKING (CHD) OF ALCOHOL IS A KNOWN RISK FACTOR FOR INCREASED SUSCEPTIBILITY TO BACTERIAL AND VIRAL INFECTION AS WELL AS IMPAIRED WOUND HEALING. EVIDENCE SUGGESTS THAT THESE DEFECTS ARE MEDIATED BY A DYSREGULATED INFLAMMATORY RESPONSE ORIGINATING FROM MYELOID CELLS, NOTABLY MONOCYTES AND MACROPHAGES, BUT THE MECHANISMS REMAIN POORLY UNDERSTOOD. OUR ABILITY TO STUDY CHD IS IMPACTED BY THE COMPLEXITIES OF HUMAN DRINKING PATTERNS AND BEHAVIOR AS WELL AS COMORBIDITIES AND CONFOUNDING RISK FACTORS FOR PATIENTS WITH ALCOHOL USE DISORDERS. TO OVERCOME THESE CHALLENGES, WE UTILIZED A TRANSLATIONAL RHESUS MACAQUE MODEL OF VOLUNTARY ETHANOL SELF-ADMINISTRATION THAT CLOSELY RECAPITULATES HUMAN DRINKING PATTERNS AND CHRONICITY. IN THIS STUDY, WE EXAMINED THE EFFECTS OF CHD ON BLOOD MONOCYTES IN CONTROL AND CHD FEMALE MACAQUES AFTER 12 MONTHS OF DAILY ETHANOL CONSUMPTION. WHILE MONOCYTES FROM CHD FEMALE MACAQUES GENERATED A HYPER-INFLAMMATORY RESPONSE TO EX VIVO LPS STIMULATION, THEIR RESPONSE TO E. COLI WAS DAMPENED. IN DEPTH SCRNA-SEQ ANALYSIS OF PURIFIED MONOCYTES REVEALED SIGNIFICANT SHIFTS IN CLASSICAL MONOCYTE SUBSETS WITH ACCUMULATION OF CELLS EXPRESSING MARKERS OF HYPOXIA (HIF1A) AND INFLAMMATION (NFKB SIGNALING PATHWAY) IN CHD MACAQUES. THE INCREASED PRESENCE OF MONOCYTE SUBSETS SKEWED TOWARDS INFLAMMATORY PHENOTYPES WAS COMPLEMENTED BY EPIGENETIC ANALYSIS, WHICH REVEALED HIGHER ACCESSIBILITY OF PROMOTER REGIONS THAT REGULATE GENES INVOLVED IN CYTOKINE SIGNALING PATHWAYS. COLLECTIVELY, DATA PRESENTED IN THIS MANUSCRIPT DEMONSTRATE THAT CHD SHIFTS CLASSICAL MONOCYTE SUBSET COMPOSITION AND PRIMES THE MONOCYTES TOWARDS A MORE HYPER-INFLAMMATORY RESPONSE TO LPS, BUT COMPROMISED PATHOGEN RESPONSE. 2021 20 6293 24 THE PRO- AND ANTI-INFLAMMATORY POTENTIAL OF IL-12: THE DUAL ROLE OF TH1 CELLS. THE DIFFERENTIATION OF T-HELPER (TH) LYMPHOCYTES INTO VARIOUS TYPES OF T-HELPER EFFECTOR AND MEMORY CELLS WITH DISTINCT FUNCTIONS DEPENDING ON THE TYPE OF CONCOMITANT SIGNALS THEY RECEIVE UPON ACTIVATION IS A CRITICAL EVENT DETERMINING THE COURSE OF AN IMMUNE REACTION. TH1 CELLS CHARACTERIZED BY THE EXPRESSION OF IFN-GAMMA AND THE RECENTLY DESCRIBED TH17 CELLS PROMOTE INFLAMMATION AND ARE CRITICALLY INVOLVED IN THE INDUCTION AND MAINTENANCE OF AUTOIMMUNITY, WHEREAS THE SECRETION OF IL-4 IS A HALLMARK OF TH2 CELLS MEDIATING PROTECTION FROM PARASITES AND ALLERGY. ORIGINAL STIMULATION IN THE PRESENCE OF IL-12 RESULTS IN THE IMPRINTING OF TH1 MEMORY CELLS FOR THE EXPRESSION OF IFN-GAMMA BY EXPRESSION OF THE TRANSCRIPTION FACTOR T-BET AND EPIGENETIC MODIFICATION OF THE IFNGAMMA GENE. IT HAS BEEN DEMONSTRATED THAT TH1 CELLS ARE POTENT INDUCERS OF INFLAMMATION. HOWEVER, IN THE CHRONIC PHASE OF SUCH INFLAMMATION, THE REGULATORY POTENTIAL OF IL-12 AND TH1 CELLS THEMSELVES MAY PLAY AN IMPORTANT ROLE IN LIMITING IMMUNOPATHOLOGY. 2007