1 4570 126 MYELOMONOCYTIC CELLS IN GIANT CELL ARTERITIS ACTIVATE TRAINED IMMUNITY PROGRAMS SUSTAINING INFLAMMATION AND CYTOKINE PRODUCTION. OBJECTIVE: TRAINED IMMUNITY (TI) IS A DE FACTO MEMORY PROGRAM OF INNATE IMMUNE CELLS, CHARACTERIZED BY IMMUNOMETABOLIC AND EPIGENETIC CHANGES SUSTAINING ENHANCED PRODUCTION OF CYTOKINES. TI EVOLVED AS A PROTECTIVE MECHANISM AGAINST INFECTIONS; HOWEVER, INAPPROPRIATE ACTIVATION CAN CAUSE DETRIMENTAL INFLAMMATION AND MIGHT BE IMPLICATED IN THE PATHOGENESIS OF CHRONIC INFLAMMATORY DISEASES. IN THIS STUDY, WE INVESTIGATED THE ROLE OF TI IN THE PATHOGENESIS OF GIANT CELL ARTERITIS (GCA), A LARGE-VESSEL VASCULITIS CHARACTERIZED BY ABERRANT MACROPHAGE ACTIVATION AND EXCESS CYTOKINE PRODUCTION. METHODS: MONOCYTES FROM GCA PATIENTS AND FROM AGE- AND SEX-MATCHED HEALTHY DONORS WERE SUBJECTED TO POLYFUNCTIONAL STUDIES, INCLUDING CYTOKINE PRODUCTION ASSAYS AT BASELINE AND FOLLOWING STIMULATION, INTRACELLULAR METABOLOMICS, CHROMATIN IMMUNOPRECIPITATION-QPCR, AND COMBINED ATAC/RNA SEQUENCING. IMMUNOMETABOLIC ACTIVATION (I.E. GLYCOLYSIS) WAS ASSESSED IN INFLAMED VESSELS OF GCA PATIENTS WITH FDG-PET AND IMMUNOHISTOCHEMISTRY (IHC), AND THE ROLE OF THIS PATHWAY IN SUSTAINING CYTOKINE PRODUCTION WAS CONFIRMED WITH SELECTIVE PHARMACOLOGIC INHIBITION IN GCA MONOCYTES. RESULTS: GCA MONOCYTES EXHIBITED HALLMARK MOLECULAR FEATURES OF TI. SPECIFICALLY, THESE INCLUDED ENHANCED IL-6 PRODUCTION UPON STIMULATION, TYPICAL IMMUNOMETABOLIC CHANGES (E.G. INCREASED GLYCOLYSIS AND GLUTAMINOLYSIS) AND EPIGENETIC CHANGES PROMOTING ENHANCED TRANSCRIPTION OF GENES GOVERNING PRO-INFLAMMATORY ACTIVATION. IMMUNOMETABOLIC CHANGES OF TI (I.E. GLYCOLYSIS) WERE A FEATURE OF MYELOMONOCYTIC CELLS IN GCA LESIONS AND WERE REQUIRED FOR ENHANCED CYTOKINE PRODUCTION. CONCLUSIONS: MYELOMONOCYTIC CELLS IN GCA ACTIVATE TI PROGRAMS SUSTAINING ENHANCED INFLAMMATORY ACTIVATION WITH EXCESS CYTOKINE PRODUCTION. 2023 2 6493 42 TRAINED IMMUNITY AND REACTIVITY OF MACROPHAGES AND ENDOTHELIAL CELLS. INNATE IMMUNE CELLS CAN DEVELOP EXACERBATED IMMUNOLOGIC RESPONSE AND LONG-TERM INFLAMMATORY PHENOTYPE FOLLOWING BRIEF EXPOSURE TO ENDOGENOUS OR EXOGENOUS INSULTS, WHICH LEADS TO AN ALTERED RESPONSE TOWARDS A SECOND CHALLENGE AFTER THE RETURN TO A NONACTIVATED STATE. THIS PHENOMENON IS KNOWN AS TRAINED IMMUNITY (TI). TI IS NOT ONLY IMPORTANT FOR HOST DEFENSE AND VACCINE RESPONSE BUT ALSO FOR CHRONIC INFLAMMATIONS SUCH AS CARDIOVASCULAR AND METABOLIC DISEASES SUCH AS ATHEROSCLEROSIS. TI CAN OCCUR IN INNATE IMMUNE CELLS SUCH AS MONOCYTES/MACROPHAGES, NATURAL KILLER CELLS, ENDOTHELIAL CELLS (ECS), AND NONIMMUNE CELLS, SUCH AS FIBROBLAST. IN THIS BRIEF REVIEW, WE ANALYZE THE SIGNIFICANCE OF TI IN ECS, WHICH ARE ALSO CONSIDERED AS INNATE IMMUNE CELLS IN ADDITION TO MACROPHAGES. TI CAN BE INDUCED BY A VARIETY OF STIMULI, INCLUDING LIPOPOLYSACCHARIDES, BCG (BACILLUS CALMETTE-GUERIN), AND OXLDL (OXIDIZED LOW-DENSITY LIPOPROTEIN), WHICH ARE DEFINED AS RISK FACTORS FOR CARDIOVASCULAR AND METABOLIC DISEASES. FURTHERMORE, TI IN ECS IS FUNCTIONAL FOR INFLAMMATION EFFECTIVENESS AND TRANSITION TO CHRONIC INFLAMMATION. REWIRING OF CELLULAR METABOLISM OF THE TRAINED CELLS TAKES PLACE DURING INDUCTION OF TI, INCLUDING INCREASED GLYCOLYSIS, GLUTAMINOLYSIS, INCREASED ACCUMULATION OF TRICARBOXYLIC ACID CYCLE METABOLITES AND ACETYL-COENZYME A PRODUCTION, AS WELL AS INCREASED MEVALONATE SYNTHESIS. SUBSEQUENTLY, THIS LEADS TO EPIGENETIC REMODELING, RESULTING IN IMPORTANT CHANGES IN CHROMATIN ARCHITECTURE THAT ENABLES INCREASED GENE TRANSCRIPTION AND ENHANCED PROINFLAMMATORY IMMUNE RESPONSE. HOWEVER, TI PATHWAYS AND INFLAMMATORY PATHWAYS ARE SEPARATED TO ENSURE MEMORY STAYS WHEN INFLAMMATION UNDERGOES RESOLUTION. ADDITIONALLY, REACTIVE OXYGEN SPECIES PLAY CONTEXT-DEPENDENT ROLES IN TI. THEREFORE, TI PLAYS SIGNIFICANT ROLES IN EC AND MACROPHAGE PATHOLOGY AND CHRONIC INFLAMMATION. HOWEVER, FURTHER CHARACTERIZATION OF TI IN ECS AND MACROPHAGES WOULD PROVIDE NOVEL INSIGHTS INTO CARDIOVASCULAR DISEASE PATHOGENESIS AND NEW THERAPEUTIC TARGETS. GRAPHIC ABSTRACT: A GRAPHIC ABSTRACT IS AVAILABLE FOR THIS ARTICLE. 2021 3 194 32 ACETYLSHIKONIN SUPPRESSES INVASION OF PORPHYROMONAS GINGIVALIS?INFECTED YD10B ORAL CANCER CELLS BY MODULATING THE INTERLEUKIN-8/MATRIX METALLOPROTEINASE AXIS. THE DEVELOPMENT OF PHARMACEUTICAL AGENTS POSSESSING ANTI?INVASIVE AND ANTI?METASTATIC ABILITIES, AS WELL AS APOPTOTIC ACTIVITY, IS IMPORTANT IN DECREASING THE INCIDENCE AND RECURRENCE OF ORAL CANCER. CANCER CELLS ARE KNOWN TO ACQUIRE INVASIVENESS NOT ONLY THROUGH EPIGENETIC CHANGES, BUT ALSO FROM INFLAMMATORY STIMULI WITHIN THE TUMOR MICROENVIRONMENT. ACCORDINGLY, THE IDENTIFICATION OF AGENTS THAT CAN SUPPRESS THE INFLAMMATION?PROMOTED INVASIVENESS OF CANCER CELLS MAY BE IMPORTANT IN TREATING CANCER AND IMPROVING THE PROGNOSIS OF PATIENTS WITH CANCER. ACETYLSHIKONIN, A FLAVONOID WITH ANTI?INFLAMMATORY ACTIVITY, INHIBITS PROLIFERATION AND INDUCES APOPTOSIS OF ORAL CANCER CELLS. IN THE PRESENT STUDY, THE ANTI?INVASIVE EFFECT OF ACETYLSHIKONIN ON YD10B ORAL CANCER CELLS INFECTED WITH PORPHYROMONAS GINGIVALIS, A MAJOR PATHOGEN OF CHRONIC PERIODONTITIS, AND THE MECHANISMS INVOLVED WERE INVESTIGATED. FIRSTLY, WE EXAMINED WHETHER P. GINGIVALIS INFECTION INCREASED THE INVASIVENESS OF YD10B CELLS. RESULTS SUGGESTED THAT YD10B ORAL CANCER CELLS BECOME MORE AGGRESSIVE WHEN THEY ARE INFECTED WITH P. GINGIVALIS. SECONDLY, ACETYLSHIKONIN SIGNIFICANTLY INHIBITED THE INVASION OF P. GINGIVALIS?INFECTED YD10B CELLS BY SUPPRESSING IL?8 RELEASE AND IL?8?DEPENDENT MMP RELEASE. THESE DATA SUGGEST THAT ACETYLSHIKONIN MAY BE A USEFUL PREVENTIVE AND THERAPEUTIC CANDIDATE FOR ORAL CANCER THAT IS CHRONICALLY INFECTED WITH PERIODONTAL PATHOGENS. 2018 4 4042 46 MACROPHAGES ACQUIRE A TNF-DEPENDENT INFLAMMATORY MEMORY IN ALLERGIC ASTHMA. BACKGROUND: INFECTIOUS AGENTS CAN REPROGRAM OR "TRAIN" MACROPHAGES AND THEIR PROGENITORS TO RESPOND MORE READILY TO SUBSEQUENT INSULTS. HOWEVER, WHETHER SUCH AN INFLAMMATORY MEMORY EXISTS IN TYPE 2 INFLAMMATORY CONDITIONS SUCH AS ALLERGIC ASTHMA WAS NOT KNOWN. OBJECTIVE: WE SOUGHT TO DECIPHER MACROPHAGE-TRAINED IMMUNITY IN ALLERGIC ASTHMA. METHODS: WE USED A COMBINATION OF CLINICAL SAMPLING OF HOUSE DUST MITE (HDM)-ALLERGIC PATIENTS, HDM-INDUCED ALLERGIC AIRWAY INFLAMMATION IN MICE, AND AN IN VITRO TRAINING SETUP TO ANALYZE PERSISTENT CHANGES IN MACROPHAGE EICOSANOID, CYTOKINE, AND CHEMOKINE PRODUCTION AS WELL AS THE UNDERLYING METABOLIC AND EPIGENETIC MECHANISMS. TRANSCRIPTIONAL AND METABOLIC PROFILES OF PATIENT-DERIVED AND IN VITRO TRAINED MACROPHAGES WERE ASSESSED BY RNA SEQUENCING OR METABOLIC FLUX ANALYSIS AND LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY ANALYSIS, RESPECTIVELY. RESULTS: WE FOUND THAT MACROPHAGES DIFFERENTIATED FROM BONE MARROW OR BLOOD MONOCYTE PROGENITORS OF HDM-ALLERGIC MICE OR ASTHMA PATIENTS SHOW INFLAMMATORY TRANSCRIPTIONAL REPROGRAMMING AND EXCESSIVE MEDIATOR (TNF-ALPHA, CCL17, LEUKOTRIENE, PGE(2), IL-6) RESPONSES UPON STIMULATION. MACROPHAGES FROM HDM-ALLERGIC MICE INITIALLY EXHIBITED A TYPE 2 IMPRINT, WHICH SHIFTED TOWARD A CLASSICAL INFLAMMATORY TRAINING OVER TIME. HDM-INDUCED ALLERGIC AIRWAY INFLAMMATION ELICITED A METABOLICALLY ACTIVATED MACROPHAGE PHENOTYPE, PRODUCING HIGH AMOUNTS OF 2-HYDROXYGLUTARATE (2-HG). HDM-INDUCED MACROPHAGE TRAINING IN VITRO WAS MEDIATED BY A FORMYL PEPTIDE RECEPTOR 2-TNF-2-HG-PGE(2)/PGE(2) RECEPTOR 2 AXIS, RESULTING IN AN M2-LIKE MACROPHAGE PHENOTYPE WITH HIGH CCL17 PRODUCTION. TNF BLOCKADE BY ETANERCEPT OR GENETIC ABLATION OF TNF IN MYELOID CELLS PREVENTED THE INFLAMMATORY IMPRINTING OF BONE MARROW-DERIVED MACROPHAGES FROM HDM-ALLERGIC MICE. CONCLUSION: ALLERGEN-TRIGGERED INFLAMMATION DRIVES A TNF-DEPENDENT INNATE MEMORY, WHICH MAY PERPETUATE AND EXACERBATE CHRONIC TYPE 2 AIRWAY INFLAMMATION AND THUS REPRESENTS A TARGET FOR ASTHMA THERAPY. 2022 5 3525 29 IL-1BETA, IL-8, AND MATRIX METALLOPROTEINASES-1, -2, AND -10 ARE ENRICHED UPON MONOCYTE-BREAST CANCER CELL COCULTIVATION IN A MATRIGEL-BASED THREE-DIMENSIONAL SYSTEM. BREAST CANCER REMAINS THE FIRST CANCER-RELATED CAUSE OF DEATH IN WOMEN WORLDWIDE, PARTICULARLY IN DEVELOPING COUNTRIES IN WHICH MOST CASES ARE DIAGNOSED IN LATE STAGES. ALTHOUGH MOST CANCER STUDIES ARE BASED IN THE GENETIC OR EPIGENETIC CHANGES OF THE TUMOR CELLS, IMMUNE CELLS WITHIN THE TUMOR STROMA OFTEN COOPERATE WITH CANCER PROGRESSION. PARTICULARLY, MONOCYTES ARE ATTRACTED TO THE TUMOR PRIMARY SITE IN WHICH THEY ARE DIFFERENTIATED INTO TUMOR-ASSOCIATED MACROPHAGES THAT FACILITATE TUMOR CELL INVASION AND METASTASIS. IN THIS STUDY, WE USED THREE-DIMENSIONAL CULTURES TO FORM ACINI-LIKE STRUCTURES TO ANALYZE THE INFLAMMATORY SECRETION PROFILE OF TUMOR CELLS INDIVIDUALLY OR IN CO-CULTURE WITH MONOCYTES. BREAST CANCER CELL LINES AND PRIMARY ISOLATES FROM EIGHT MEXICAN PATIENTS WITH BREAST CANCER WERE USED. WE FOUND HIGH LEVELS OF RANTES/CCL5, MCP-1/CCL2, AND G-CSF IN THE BREAST CANCER INDIVIDUAL CULTURES, SUPPORTING AN IMPORTANT RECRUITMENT CAPACITY OF MONOCYTES, BUT ALSO OF NEUTROPHILS. THE CO-CULTURES OF THE TUMOR CELLS AND MONOCYTES WERE SIGNIFICANTLY ENRICHED WITH THE POTENT PRO-INFLAMMATORY CYTOKINES INTERLEUKIN (IL)-1BETA AND IL-8, KNOWN TO SUPPORT MALIGNANT PROGRESSION. WE ALSO FOUND THAT THE INTERACTION OF TUMOR CELLS WITH MONOCYTES PROMOTED HIGH LEVELS OF MATRIX METALLOPROTEINASES (MMP)-1, MMP-2, AND MMP-10. OUR STUDY SUPPORTS THAT A KEY EVENT FOR MALIGNANT PROGRESSION IS THE RECRUITMENT OF DIFFERENT IMMUNE CELL POPULATIONS, WHICH HELP TO SUSTAIN AND ENHANCE A CHRONIC INFLAMMATORY MICROENVIRONMENT THAT HIGHLY FAVORS TUMOR MALIGNANCY. 2017 6 1594 34 DNA METHYLATION PROFILING REVEALS DIFFERENCES IN THE 3 HUMAN MONOCYTE SUBSETS AND IDENTIFIES UREMIA TO INDUCE DNA METHYLATION CHANGES DURING DIFFERENTIATION. HUMAN MONOCYTES ARE A HETEROGENEOUS CELL POPULATION CONSISTING OF 3 SUBSETS: CLASSICAL CD14++CD16-, INTERMEDIATE CD14++CD16+ AND NONCLASSICAL CD14+CD16++ MONOCYTES. VIA POORLY CHARACTERIZED MECHANISMS, INTERMEDIATE MONOCYTE COUNTS RISE IN CHRONIC INFLAMMATORY DISEASES, AMONG WHICH CHRONIC KIDNEY DISEASE IS OF PARTICULAR EPIDEMIOLOGIC IMPORTANCE. DNA METHYLATION IS A CENTRAL EPIGENETIC FEATURE THAT CONTROLS HEMATOPOIESIS. BY APPLYING NEXT-GENERATION METHYL-SEQUENCING WE NOW TESTED HOW FAR THE 3 MONOCYTE SUBSETS DIFFER IN THEIR DNA METHYLOME AND WHETHER UREMIA INDUCES DNA METHYLATION CHANGES IN DIFFERENTIATING MONOCYTES. WE FOUND THAT EACH MONOCYTE SUBSET DISPLAYS A UNIQUE PHENOTYPE WITH REGARDS TO DNA METHYLATION. GENES WITH DIFFERENTIALLY METHYLATED PROMOTER REGIONS IN INTERMEDIATE MONOCYTES WERE LINKED TO DISTINCT IMMUNOLOGICAL PROCESSES, WHICH IS IN LINE WITH RESULTS FROM RECENT GENE EXPRESSION ANALYSES. IN VITRO, UREMIA INDUCED DYSREGULATION OF DNA METHYLATION IN DIFFERENTIATING MONOCYTES, WHICH AFFECTED SEVERAL TRANSCRIPTION REGULATORS IMPORTANT FOR MONOCYTE DIFFERENTIATION (E.G., FLT3, HDAC1, MNT) AND LED TO ENHANCED GENERATION OF INTERMEDIATE MONOCYTES. AS POTENTIAL MEDIATOR, THE UREMIC TOXIN AND METHYLATION INHIBITOR S-ADENOSYLHOMOCYSTEINE INDUCED SHIFTS IN MONOCYTE SUBSETS IN VITRO, AND ASSOCIATED WITH MONOCYTE SUBSET COUNTS IN VIVO. OUR DATA SUPPORT THE CONCEPT OF MONOCYTE TRICHOTOMY AND THE DISTINCT ROLE OF INTERMEDIATE MONOCYTES IN HUMAN IMMUNITY. THE SHIFT IN MONOCYTE SUBSETS THAT OCCURS IN CHRONIC KIDNEY DISEASE, A PROINFLAMMATORY CONDITION OF SUBSTANTIAL EPIDEMIOLOGICAL IMPACT, MAY BE INDUCED BY ACCUMULATION OF UREMIC TOXINS THAT MEDIATE EPIGENETIC DYSREGULATION. 2016 7 5088 32 PIPERLONGUMINE REGULATES EPIGENETIC MODULATION AND ALLEVIATES PSORIASIS-LIKE SKIN INFLAMMATION VIA INHIBITION OF HYPERPROLIFERATION AND INFLAMMATION. PSORIASIS IS AN AUTOIMMUNE SKIN DISEASE, WHERE CHRONIC IMMUNE RESPONSES DUE TO EXAGGERATED CYTOKINE SIGNALING, ABNORMAL DIFFERENTIATION, AND EVASION OF KERATINOCYTES APOPTOSIS PLAYS A CRUCIAL ROLE IN MEDIATING ABNORMAL KERATINOCYTES HYPERPROLIFERATION. FROM THE THERAPEUTIC PERSPECTIVE, THE MOLECULES WITH STRONG ANTI-PROLIFERATIVE AND ANTI-INFLAMMATORY PROPERTIES COULD HAVE TREMENDOUS RELEVANCE. IN THIS STUDY, WE DEMONSTRATED THAT PIPERLONGUMINE (PPL) TREATMENT EFFECTIVELY ABROGATED THE HYPERPROLIFERATION AND DIFFERENTIATION OF KERATINOCYTES BY INDUCING ROS-MEDIATED LATE APOPTOSIS WITH LOSS OF MITOCHONDRIAL MEMBRANE POTENTIAL. BESIDES, THE ARREST OF CELL CYCLE WAS FOUND AT SUB-G1 PHASE AS A RESULT OF DNA FRAGMENTATION. MOLECULARLY, INHIBITION OF STAT3 AND AKT SIGNALING WAS OBSERVED WITH A DECREASE IN PROLIFERATIVE MARKERS SUCH AS PCNA, KI67, AND CYCLIN D1 ALONG WITH ANTI-APOPTOTIC BCL-2 PROTEIN EXPRESSION. KERATIN 17 IS A CRITICAL REGULATOR OF KERATINOCYTE DIFFERENTIATION, AND IT WAS FOUND TO BE DOWNREGULATED WITH PPL SIGNIFICANTLY. FURTHERMORE, PROMINENT ANTI-INFLAMMATORY EFFECTS WERE OBSERVED BY INHIBITION OF LIPOPOLYSACCHARIDE (LPS)/IMIQUIMOD (IMQ)-INDUCED P65 NF-KAPPAB SIGNALING CASCADE AND STRONGLY INHIBITED THE PRODUCTION OF CYTOKINE STORM INVOLVED IN PSORIASIS-LIKE SKIN INFLAMMATION, THUS LED TO THE RESTORATION OF NORMAL EPIDERMAL ARCHITECTURE WITH REDUCTION OF EPIDERMAL HYPERPLASIA AND SPLENOMEGALY. IN ADDITION, PPL EPIGENETICALLY INHIBITED HISTONE-MODIFYING ENZYMES, WHICH INCLUDE HISTONE DEACETYLASES (HDACS) OF CLASS I (HDAC1-4) AND CLASS II (HDAC6) EVALUATED BY IMMUNOBLOTTING AND HDAC ENZYME ASSAY KIT. IN ADDITION, OUR RESULTS SHOW THAT PPL EFFECTIVELY INHIBITS THE NUCLEAR TRANSLOCATION OF P65 AND A HISTONE MODULATOR HDAC3, THUS SEQUESTERED IN THE CYTOPLASM OF MACROPHAGES. FURTHERMORE, PPL EFFECTIVELY ENHANCED THE PROTEIN-PROTEIN INTERACTIONS OF HDAC3 AND P65 WITH IKAPPABALPHA, WHICH WAS DISRUPTED BY LPS STIMULATION AND WERE EVALUATED BY CO-IP AND MOLECULAR MODELING. COLLECTIVELY, OUR FINDINGS INDICATE THAT PIPERLONGUMINE MAY SERVE AS AN ANTI-PROLIFERATIVE AND ANTI-INFLAMMATORY AGENT AND COULD SERVE AS A POTENTIAL THERAPEUTIC OPTION IN TREATING PSORIASIS. 2020 8 4210 27 METFORMIN AND VITAMIN D MODULATE INFLAMMATION AND AUTOPHAGY DURING ADIPOSE-DERIVED STEM CELL DIFFERENTIATION. ADIPOSE-DERIVED STEM CELLS (ADSCS) CAME OUT FROM THE REGENERATIVE MEDICINE LANDSCAPE FOR THEIR ABILITY TO DIFFERENTIATE INTO SEVERAL PHENOTYPES, CONTRIBUTING TO TISSUE REGENERATION BOTH IN VITRO AND IN VIVO. DYSREGULATION IN STEM CELL RECRUITMENT AND DIFFERENTIATION DURING ADIPOGENESIS IS LINKED TO A CHRONIC LOW-GRADE INFLAMMATION AND MACROPHAGE INFILTRATION INSIDE THE ADIPOSE TISSUE, INSULIN RESISTANCE, CARDIOVASCULAR DISEASE AND OBESITY. IN THE PRESENT PAPER WE AIMED TO EVALUATE THE ROLE OF METFORMIN AND VITAMIN D, ALONE OR IN COMBINATION, IN MODULATING INFLAMMATION AND AUTOPHAGY IN ADSCS DURING ADIPOGENIC COMMITMENT. ADSCS WERE CULTURED FOR 21 DAYS IN THE PRESENCE OF A SPECIFIC ADIPOGENIC DIFFERENTIATION MEDIUM, TOGETHER WITH METFORMIN, OR VITAMIN D, OR BOTH. WE THEN ANALYZED THE EXPRESSION OF FOXO1 AND HEAT SHOCK PROTEINS (HSP) AND THE SECRETION OF PROINFLAMMATORY CYTOKINES IL-6 AND TNF-ALPHA BY ELISA. AUTOPHAGY WAS ALSO ASSESSED BY SPECIFIC WESTERN BLOT ANALYSIS OF ATG12, LC3B I, AND LC3B II EXPRESSION. OUR RESULTS SHOWED THE ABILITY OF THE CONDITIONED MEDIA TO MODULATE ADIPOGENIC DIFFERENTIATION, FINELY TUNING THE INFLAMMATORY RESPONSE AND AUTOPHAGY. WE OBSERVED A MODULATION IN HSP MRNA LEVELS, AND A SIGNIFICANT DOWNREGULATION IN CYTOKINE SECRETION. TAKEN TOGETHER, OUR FINDINGS SUGGEST THE POSSIBLE APPLICATION OF THESE MOLECULES IN CLINICAL PRACTICE TO COUNTERACT UNCONTROLLED LIPOGENESIS AND PREVENT OBESITY AND OBESITY-RELATED METABOLIC DISORDERS. 2021 9 3893 28 LACTATE INDUCES METABOLIC AND EPIGENETIC REPROGRAMMING OF PRO-INFLAMMATORY TH17 CELLS. INCREASED LACTATE LEVELS IN THE TISSUE MICROENVIRONMENT ARE A WELL-KNOWN FEATURE OF CHRONIC INFLAMMATION. HOWEVER, THE ROLE OF LACTATE IN REGULATING T CELL FUNCTION REMAINS CONTROVERSIAL. HERE, WE DEMONSTRATE THAT EXTRACELLULAR LACTATE PREDOMINANTLY INDUCES DEREGULATION OF THE TH17-SPECIFIC GENE EXPRESSION PROGRAM BY MODULATING THE METABOLIC AND EPIGENETIC STATUS OF TH17 CELLS. FOLLOWING LACTATE TREATMENT, TH17 CELLS SIGNIFICANTLY REDUCED THEIR IL-17A PRODUCTION AND UPREGULATED FOXP3 EXPRESSION THROUGH ROS-DRIVEN IL-2 SECRETION. MOREOVER, WE OBSERVED INCREASED LEVELS OF GENOME-WIDE HISTONE H3K18 LACTYLATION, A RECENTLY DESCRIBED MARKER FOR ACTIVE CHROMATIN IN MACROPHAGES, IN LACTATE-TREATED TH17 CELLS. IN ADDITION, WE SHOW THAT HIGH LACTATE CONCENTRATIONS SUPPRESS TH17 PATHOGENICITY DURING INTESTINAL INFLAMMATION IN MICE. THESE RESULTS INDICATE THAT LACTATE IS CAPABLE OF REPROGRAMMING PRO-INFLAMMATORY T CELL PHENOTYPES INTO REGULATORY T CELLS. 2022 10 2340 37 EPIGENETIC REGULATION OF LEUKOCYTE INFLAMMATORY MEDIATOR PRODUCTION DICTATES STAPHYLOCOCCUS AUREUS CRANIOTOMY INFECTION OUTCOME. STAPHYLOCOCCUS AUREUS IS A COMMON CAUSE OF SURGICAL-SITE INFECTIONS, INCLUDING THOSE ARISING AFTER CRANIOTOMY, WHICH IS PERFORMED TO ACCESS THE BRAIN FOR THE TREATMENT OF TUMORS, EPILEPSY, OR HEMORRHAGE. CRANIOTOMY INFECTION IS CHARACTERIZED BY COMPLEX SPATIAL AND TEMPORAL DYNAMICS OF LEUKOCYTE RECRUITMENT AND MICROGLIAL ACTIVATION. WE RECENTLY IDENTIFIED UNIQUE TRANSCRIPTIONAL PROFILES OF THESE IMMUNE POPULATIONS DURING S. AUREUS CRANIOTOMY INFECTION. EPIGENETIC PROCESSES ALLOW RAPID AND REVERSIBLE CONTROL OVER GENE TRANSCRIPTION; HOWEVER, LITTLE IS KNOWN ABOUT HOW EPIGENETIC PATHWAYS INFLUENCE IMMUNITY TO LIVE S. AUREUS. AN EPIGENETIC COMPOUND LIBRARY SCREEN IDENTIFIED BROMODOMAIN AND EXTRATERMINAL DOMAIN-CONTAINING (BET) PROTEINS AND HISTONE DEACETYLASES (HDACS) AS CRITICAL FOR REGULATING TNF, IL-6, IL-10, AND CCL2 PRODUCTION BY PRIMARY MOUSE MICROGLIA, MACROPHAGES, NEUTROPHILS, AND GRANULOCYTIC MYELOID-DERIVED SUPPRESSOR CELLS IN RESPONSE TO LIVE S. AUREUS. CLASS I HDACS (C1HDACS) WERE INCREASED IN THESE CELL TYPES IN VITRO AND IN VIVO DURING ACUTE DISEASE IN A MOUSE MODEL OF S. AUREUS CRANIOTOMY INFECTION. HOWEVER, SUBSTANTIAL REDUCTIONS IN C1HDACS WERE OBSERVED DURING CHRONIC INFECTION, HIGHLIGHTING TEMPORAL REGULATION AND THE IMPORTANCE OF THE TISSUE MICROENVIRONMENT FOR DICTATING C1HDAC EXPRESSION. MICROPARTICLE DELIVERY OF HDAC AND BET INHIBITORS IN VIVO CAUSED WIDESPREAD DECREASES IN INFLAMMATORY MEDIATOR PRODUCTION, WHICH SIGNIFICANTLY INCREASED BACTERIAL BURDEN IN THE BRAIN, GALEA, AND BONE FLAP. THESE FINDINGS IDENTIFY HISTONE ACETYLATION AS AN IMPORTANT MECHANISM FOR REGULATING CYTOKINE AND CHEMOKINE PRODUCTION ACROSS DIVERSE IMMUNE CELL LINEAGES THAT IS CRITICAL FOR BACTERIAL CONTAINMENT. ACCORDINGLY, ABERRANT EPIGENETIC REGULATION MAY BE IMPORTANT FOR PROMOTING S. AUREUS PERSISTENCE DURING CRANIOTOMY INFECTION. 2023 11 5114 30 PORPHYROMONAS GINGIVALIS LIPOPOLYSACCHARIDE STIMULATION IN HUMAN PERIODONTAL LIGAMENT STEM CELLS: ROLE OF EPIGENETIC MODIFICATIONS TO THE INFLAMMATION. PERIODONTITIS IS A CHRONIC ORAL INFLAMMATORY DISEASE PRODUCED BY BACTERIA. GINGIVAL RETRACTION AND BONE AND CONNECTIVE TISSUES RESORPTION ARE THE HALLMARKS OF THIS DISEASE. CHRONIC PERIODONTITIS MAY CONTRIBUTE TO THE RISK OF ONSET OR PROGRESSION OF NEUROINFLAMMATORY PATHOLOGICAL CONDITIONS, SUCH AS ALZHEIMER'S DISEASE. THE MAIN GOAL OF THE PRESENT STUDY WAS TO INVESTIGATE IF THE ROLE OF EPIGENETIC MODULATIONS IS INVOLVED IN PERIODONTITIS USING HUMAN PERIODONTAL LIGAMENT STEM CELLS (HPDLSCS) AS AN IN VITRO MODEL SYSTEM. HPDLSCS WERE TREATED WITH LIPOPOLYSACCHARIDE OF PORPHYROMONAS GINGIVALIS AND THE EXPRESSION OF PROTEINS ASSOCIATED WITH DNA METHYLATION AND HISTONE ACETYLATION, SUCH AS DNMT1 AND P300, RESPECTIVELY, AND INFLAMMATORY TRANSCRIPTION FACTOR NF-KB, WERE EXAMINED. IMMUNOFLUORESCENCE, WESTERN BLOT AND NEXT GENERATION SEQUENCING RESULTS DEMONSTRATED THAT P. GINGIVALIS LIPOPOLYSACCHARIDE SIGNIFICANTLY REDUCED DNA METHYLASE DNMT1, WHILE IT MARKEDLY UPREGULATED THE LEVEL OF HISTONE ACETYLTRANSFERASE P300 AND NF-KB IN HPDLSCS. OUR RESULTS SHOWED THAT P. GINGIVALIS LIPOPOLYSACCHARIDE MARKEDLY REGULATE THE GENES INVOLVED IN EPIGENETIC MECHANISM, WHICH MAY RESULT IN INFLAMMATION INDUCTION. WE PROPOSE THAT P. GINGIVALIS LIPOPOLYSACCHARIDE-TREATED HPDLSCS COULD BE A POTENTIAL IN VITRO MODEL SYSTEM TO STUDY EPIGENETICS MODULATIONS ASSOCIATED WITH PERIODONTITIS, WHICH MIGHT BE HELPFUL TO IDENTIFY NOVEL BIOMARKERS LINKED TO THIS ORAL INFLAMMATORY DISEASE. 2017 12 5982 35 TET2 REGULATES IMMUNE TOLERANCE IN CHRONICALLY ACTIVATED MAST CELLS. MUTATION OF THE TET2 DNA-HYDROXYMETHYLASE HAS BEEN ASSOCIATED WITH A NUMBER OF IMMUNE PATHOLOGIES. THE DISPARITY IN PHENOTYPE AND CLINICAL PRESENTATION AMONG THESE PATHOLOGIES LEADS TO QUESTIONS REGARDING THE ROLE OF TET2 MUTATION IN PROMOTING DISEASE EVOLUTION IN DIFFERENT IMMUNE CELL TYPES. HERE WE SHOW THAT, IN PRIMARY MAST CELLS, TET2 EXPRESSION IS INDUCED IN RESPONSE TO CHRONIC AND ACUTE ACTIVATION SIGNALS. IN TET2-DEFICIENT MAST CELLS, CHRONIC ACTIVATION VIA THE ONCOGENIC KITD816V ALLELE ASSOCIATED WITH MASTOCYTOSIS, SELECTS FOR A SPECIFIC EPIGENETIC SIGNATURE CHARACTERIZED BY HYPERMETHYLATED DNA REGIONS (HMR) AT IMMUNE RESPONSE GENES. H3K27AC AND TRANSCRIPTION FACTOR BINDING IS CONSISTENT WITH PRIMING OR MORE OPEN CHROMATIN AT BOTH HMR AND NON-HMR IN PROXIMITY TO IMMUNE GENES IN THESE CELLS, AND THIS SIGNATURE COINCIDES WITH INCREASED PATHOLOGICAL INFLAMMATION SIGNALS. HMR ARE ALSO ASSOCIATED WITH A SUBSET OF IMMUNE GENES THAT ARE DIRECT TARGETS OF TET2 AND REPRESSED IN TET2-DEFICIENT CELLS. REPRESSION OF THESE GENES RESULTS IN IMMUNE TOLERANCE TO ACUTE STIMULATION THAT CAN BE RESCUED WITH VITAMIN C TREATMENT OR REITERATED WITH A TET INHIBITOR. OVERALL, OUR DATA SUPPORT A MODEL WHERE TET2 PLAYS A DIRECT ROLE IN PREVENTING IMMUNE TOLERANCE IN CHRONICALLY ACTIVATED MAST CELLS, SUPPORTING TET2 AS A VIABLE TARGET TO REPROGRAM THE INNATE IMMUNE RESPONSE FOR INNOVATIVE THERAPIES. 2022 13 19 32 5-AZACYTYDINE AND RESVERATROL REVERSE SENESCENCE AND AGEING OF ADIPOSE STEM CELLS VIA MODULATION OF MITOCHONDRIAL DYNAMICS AND AUTOPHAGY. OBESITY AND ENDOCRINE DISORDERS HAVE BECOME PREVALENT ISSUES IN THE FIELD OF BOTH HUMAN AND VETERINARY MEDICINE. EQUINE METABOLIC SYNDROME IS A COMPLEX DISORDER INVOLVING ALTERNATION IN METABOLISM AND CHRONIC SYSTEMIC INFLAMMATION. IT HAS BEEN SHOWN THAT UNFAVOURABLE MICROENVIRONMENT OF INFLAMED ADIPOSE TISSUE NEGATIVELY AFFECTS ADIPOSE STEM CELL POPULATION (ASC) RESIDING WITHIN, MARKEDLY LIMITING THEIR THERAPEUTIC POTENTIAL. ASCS(EMS) ARE CHARACTERIZED BY INCREASED SENESCENCE APOPTOSIS, EXCESSIVE ACCUMULATION OF REACTIVE OXYGEN SPECIES (ROS), MITOCHONDRIA DETERIORATION AND "AUTOPHAGIC FLUX." THE AIM OF THE PRESENT STUDY WAS TO EVALUATE WHETHER TREATMENT OF ASCS(EMS) WITH A COMBINATION OF 5-AZACYTYDINE (AZA) AND RESVERATROL (RES) WOULD REVERSE AGED PHENOTYPE OF THESE CELLS. FOR THIS REASON, WE PERFORMED THE FOLLOWING ANALYZES: MOLECULAR BIOLOGY (RT-PCR), MICROSCOPIC (IMMUNOFLUORESCENCE, TEM) AND FLOW CYTOMETRY (JC-1, ROS, KI67). WE EVALUATED THE MITOCHONDRIAL STATUS, DYNAMICS AND CLEARANCE AS WELL AS AUTOPHAGIC PATHWAYS. FURTHERMORE, WE INVESTIGATED EPIGENETIC ALTERNATIONS IN TREATED CELLS BY MEASURING THE EXPRESSION OF TET GENES AND ANALYSIS OF DNA METHYLATION STATUS. WE HAVE DEMONSTRATED THAT AZA/RES TREATMENT OF ASCS(EMS) IS ABLE TO REJUVENATE THESE CELLS BY MODULATING MITOCHONDRIAL DYNAMICS, IN PARTICULAR BY PROMOTING MITOCHONDRIAL FUSION OVER FISSION. AFTER AZA/RES TREATMENT, ASCS(EMS) WERE CHARACTERIZED BY INCREASED PROLIFERATION RATE, DECREASED APOPTOSIS AND SENESCENCE AND LOWER ROS ACCUMULATION. OUR FINDINGS OFFER A NOVEL APPROACH AND POTENTIAL TARGETS FOR THE BENEFICIAL EFFECTS OF AZA/RES IN AMELIORATING STEM CELL DYSFUNCTIONS. 2019 14 855 33 CHROMATIN ACCESSIBILITY LANDSCAPES OF IMMUNE CELLS IN RHEUMATOID ARTHRITIS NOMINATE MONOCYTES IN DISEASE PATHOGENESIS. BACKGROUND: RHEUMATOID ARTHRITIS (RA) IS A CHRONIC, SYSTEMIC AUTOIMMUNE DISEASE THAT INVOLVES A VARIETY OF CELL TYPES. HOWEVER, HOW THE EPIGENETIC DYSREGULATIONS OF PERIPHERAL IMMUNE CELLS CONTRIBUTE TO THE PATHOGENESIS OF RA STILL REMAINS LARGELY UNCLEAR. RESULTS: HERE, WE ANALYSED THE GENOME-WIDE ACTIVE DNA REGULATORY ELEMENTS OF FOUR MAJOR IMMUNE CELLS, NAMELY MONOCYTES, B CELLS, CD4(+) T CELLS AND CD8(+) T CELLS, IN PERIPHERAL BLOOD OF RA PATIENTS, OSTEOARTHRITIS (OA) PATIENTS AND HEALTHY DONORS USING ASSAY OF TRANSPOSASE ACCESSIBLE CHROMATIN WITH SEQUENCING (ATAC-SEQ). WE FOUND A STRONG RA-ASSOCIATED CHROMATIN DYSREGULATION SIGNATURE IN MONOCYTES, BUT NO OTHER EXAMINED CELL TYPES. MOREOVER, WE FOUND THAT SERUM C-REACTIVE PROTEIN (CRP) CAN INDUCE THE RA-ASSOCIATED CHROMATIN DYSREGULATION IN MONOCYTES VIA IN VITRO EXPERIMENTS. AND THE EXTENT OF THIS DYSREGULATION WAS REGULATED THROUGH THE TRANSCRIPTION FACTOR FRA2. CONCLUSIONS: TOGETHER, OUR STUDY REVEALED A CRP-INDUCED PATHOGENIC CHROMATIN DYSREGULATION SIGNATURE IN MONOCYTES FROM RA PATIENTS AND PREDICTED THE RESPONSIBLE SIGNALLING PATHWAY AS POTENTIAL THERAPEUTIC TARGETS FOR THE DISEASE. 2021 15 3948 29 LNCRNA-CD160 DECREASES THE IMMUNITY OF CD8(+) T CELLS THROUGH EPIGENETIC MECHANISMS IN HEPATITIS B VIRUS INFECTION. THE TRANSFER AND DEVELOPMENT OF CHRONIC HEPATITIS B VIRUS (HBV) INFECTION IS ASSOCIATED WITH THE T CELL IMMUNE RESPONSE, THEREFORE INVESTIGATING THE KEY REGULATORS OF CELL IMMUNE RESPONSE IS NEEDED TO IMPROVE CHRONIC HBV TREATMENT. BLOOD SAMPLES FROM PATIENTS WITH CHRONIC HBV INFECTION WERE USED TO CONFIRM THE CORRELATION BETWEEN HBV INFECTION STAGE AND CD160 RECEPTOR EXPRESSION LEVELS IN CD8(+) T CELLS, THE CD8(+) T CELLS ARE USED TO RESEARCH THE MECHANISM OF T CELL IMMUNE RESPONSE MODULATION, MOREOVER, C3H/HEN MICE WITH REDUCED CD160 EXPRESSION LEVELS WERE USED TO INVESTIGATE THE ASSOCIATION BETWEEN LONG NON-CODING (LNC)RNA-CD160 AND HBV INFECTION. LONG NON-CODING (LNC)RNA-CD160 AND HISTONE-MODIFICATION ENZYME GENE HISTONE DEACETYLASE 11 (HDAC11) EXPRESSION LEVELS WERE NEGATIVELY ASSOCIATED WITH CD160 EXPRESSION. LNCRNA-CD160 CAN INHIBIT THE SECRETION OF IFN-GAMMA AND TNF-ALPHA THROUGH HDAC11 RECRUITMENT AND BIND TO HDAC11 TO FORM A COMPLEX ON THE PROMOTERS OF IFN-GAMMA AND TNF-ALPHA. THE HDAC11, IFN-GAMMA AND TNF-ALPHA FORM A COMPLEX AND ENHANCE THE METHYLATION OF H3K9ME1, CHROMATIN CHANGES INTO THE HETEROCHROMATIN AND THE TRANSCRIPTION OF IFN-GAMMA AND TNF-ALPHA IS BLOCKED; MOREOVER, THE HDAC11/IFN-GAMMA/TNF-ALPHA COMPLEX CAN ALSO INHIBIT THE SECRETION OF IFN-GAMMA AND TNF-ALPHA IN CD160(-) CD8(+) T CELLS AND SUPPRESSES THE FUNCTION OF CD8(+) T CELLS. FURTHERMORE, SMALL INTERFERING RNA TARGETING LNCRNA-CD160 CAN BLOCK HBV INFECTION PROGRESSION. LNCRNA-CD160 ACTS AS AN IMMUNE SUPPRESSIVE FACTOR AND IS EXPRESSED AT A HIGH LEVEL IN PERIPHERAL BLOOD CD8(+) T CELLS OF HBV INFECTED PATIENTS. FURTHERMORE, HIGH EXPRESSION LEVELS OF LNCRNA-CD160 CAN CONTRIBUTE TO THE INHIBITION OF IFN-GAMMA AND TNF-ALPHA SECRETION IN CD8(+) T CELLS AND DECREASE THE IMMUNE RESPONSE OF CD8(+) T CELLS. THEREFORE, LNCRNA-CD160 MAY BECOME A NEW TARGET FOR IMMUNOTHERAPY OF CHRONIC HBV INFECTION IN THE FUTURE AND MAY PROVIDE A NEW THERAPEUTIC STRATEGY FOR THE TREATMENT OF HBV INFECTION. 2020 16 4831 46 OLIGOMERIC S100A4 IS ASSOCIATED WITH MONOCYTE INNATE IMMUNE MEMORY AND BYPASS OF TOLERANCE TO SUBSEQUENT STIMULATION WITH LIPOPOLYSACCHARIDES. OBJECTIVES: MOST DAMPS IN INFLAMMATORY DISEASES ARE TLR2- AND TLR4-LIGANDS AND ACCORDING TO THE CURRENT CONCEPT, REPEATED STIMULI WOULD RESULT IN TOLERANCE. AIMS OF THE STUDY WERE TO VERIFY THIS ASSUMPTION, TO INVESTIGATE WHETHER EPIGENETIC EFFECTORS ARE INVOLVED AND TO EXPLORE THE SITUATION IN RHEUMATOID ARTHRITIS (RA). METHODS: A TRAINED IMMUNITY (TI) AND TOLERANCE PROTOCOL WAS ESTABLISHED USING PERIPHERAL BLOOD MONOCYTES FROM HEALTHY DONORS, BETA-GLUCAN AND LIPOPOLYSACCHARIDE (LPS). THE TRAINING OR TOLERANCE CAPACITIES OF RA-RELEVANT DAMPS WERE TESTED. RESULTS: BETA-GLUCAN-, OS100A4-, HMBG1-, AND HSP90-PRETREATED MONOCYTES SHOWED INCREASED IL-6 RESPONSES TO LPS RE-STIMULATION. BETA-GLUCAN, OS100A AND TENASCIN C INDUCED TRAINING OF MONOCYTES TO RELEASE MORE TNFALPHA. IN COMPARISON TO BETA-GLUCAN, MOST DAMPS TESTED INDUCED LESS TI, WITH EXCEPTION OF OS100A4. MONOCYTES EXPOSED TO OS100A4 SHOWED INCREASED IL-1BETA, IL-6, AND TNFALPHA IN RESPONSE TO LPS, IN SPITE THAT BOTH STIMULATE TLR4. RNASEQ UPON BETA-GLUCAN OR OS100A4 REVEALED SIMILAR CHANGES IN CHEMOKINES/CYTOKINES AND EPIGENETIC EFFECTORS; 17 EPIGENETIC EFFECTORS CORRELATED WITH CHEMOKINE/CYTOKINE GENE EXPRESSION; PRDM8 WAS ASSOCIATED WITH MORE CHEMOKINE AND CYTOKINE TRANSCRIPTS. KNOCKDOWN OF PRDM8 ABOLISHED TI INDUCED BY OS100A4. IN RA, PLASMA S100A4 CORRELATED WITH INCREASED CSF2, AND INCREASED PRDM8 TRANSCRIPTION IN RA MONOCYTES WAS ASSOCIATED WITH INCREASED PLASMA CCL5 AND IL-6, AS WELL AS THERAPY-RESISTANCE. CONCLUSION: BYPASS OF TOLERANCE BY DAMPS MIGHT BE A PHENOMENON AS IMPORTANT AS TI, SINCE IT COULD EXPLAIN HOW CHRONIC INFLAMMATION CAN BE MAINTAINED IN SPITE OF AN ENVIRONMENT WITH MULTIPLE TLR2/TLR4-LIGANDS. IN RA MONOCYTES, A PRDM8-DEPENDENT TI MECHANISM COULD BE RESPONSIBLE FOR SUSTAINED CHEMOKINE/CYTOKINES LEVELS. 2019 17 5491 32 REVIEW ARTICLE: INFLAMMATION-RELATED PROMOTION OF GASTROINTESTINAL CARCINOGENESIS--A PERIGENETIC PATHWAY. CHRONIC INFLAMMATION HAS BEEN REPORTED TO ACCELERATE NEOPLASMAS IN GASTROINTESTINAL TRACT. CERTAIN BACTERIA INCLUDING HELICOBACTER PYLORI DIRECTLY INTERACT WITH HOST CELLS, INDUCE PROINFLAMMATORY CYTOKINES AND STIMULATE PRODUCTION OF FREE RADICALS. FREE RADICALS CAUSE MUTATIONS IN TARGET CELLS SO THAT NEOPLASTIC CLONES ARE ESTABLISHED. ACCUMULATION OF SUCH GENETIC ALTERATIONS MAY CAUSE MALIGNANT TRANSFORMATION OF SOME ESTABLISHED CLONES. IN ADDITION, INFLAMMATORY ALTERATIONS MAY PROMOTE GROWTH, EXPANSION AND INVASION OF GASTROINTESTINAL EPITHELIAL CELLS. THE LATTER CHANGES CAUSED BY INFLAMMATION MAY OCCUR EVEN WITHOUT FURTHER GENETIC MUTATIONS OR EPIGENETIC ALTERATIONS, AND THEREFORE MAY BE CATEGORIZED AS 'PERIGENETIC ALTERATIONS' OF NEOPLASTIC CELLS. FOR AN EXAMPLE, TUMOUR NECROSIS FACTOR ALPHA (TNF-ALPHA) PLAYS PIVOTAL ROLES NOT ONLY IN THE REDUCTION BUT ALSO IN THE GROWTH, INVASION AND METASTASES OF CERTAIN NEOPLASMAS. OUR STUDIES SHOW THAT TNF-ALPHA INCREASES INTRACELLULAR RADICAL PRODUCTION, DEGRADATES E-CADHERIN / BETA-CATENIN COMPLEX AND PROMOTES DISPERSION AND MIGRATION IN EPITHELIAL CELLS TRANSFORMED WITH AN ACTIVATED SRC ONCOGENE (V-SRC). THESE DATA INDICATE THAT AN INFLAMMATORY CYTOKINE INDUCES THE MALIGNANT POTENTIAL OF SRC-ACTIVATED NEOPLASTIC CELLS. INTERESTINGLY, TNF-ALPHA ALSO INDUCED THESE PHENOTYPIC CHANGES IN NONMUTATED CELLS WHOSE C-SRC WAS ACTIVATED BY TGF-ALPHA, SUGGESTING THAT THE INVASIVE PROPERTIES OF THE CELL WERE NOT NECESSARILY RELATED TO GENE MUTATION. FURTHERMORE, CERTAIN RADICAL SCAVENGERS SUPPRESSED THE INVASIVE PHENOTYPE OF THE CELLS. THESE RESULTS INDICATE THAT PERIGENETIC ALTERATIONS ARE AN IMPORTANT TARGET OF PHARMACOLOGICAL INTERVENTION OF CARCINOGENESIS. 2003 18 1618 37 DNA METHYLTRANSFERASE INHIBITORS INCREASE NOD-LIKE RECEPTOR ACTIVITY AND EXPRESSION IN A MONOCYTIC CELL LINE. BACKGROUND: THE INTRACELLULAR NOD-LIKE RECEPTOR (NLR) FAMILY OF PATHOGEN RECOGNITION RECEPTORS (PRRA) IS INVOLVED IN INITIATING THE INNATE IMMUNE RESPONSE OF WHICH NOD1 AND NOD2 ARE THE BEST-CHARACTERIZED MEMBERS. ABERRANT EXPRESSION OF NOD1 AND NOD2 HAS BEEN UNCOVERED IN A NUMBER OF CHRONIC INFLAMMATORY DISEASES, SUCH AS INFLAMMATORY BOWEL DISEASE AND RHEUMATOID ARTHRITIS. HOWEVER, THE MECHANISM UNDERLYING NOD1/NOD2 GENE EXPRESSION REGULATION IS STILL IN ITS INFANCY. EPIGENETIC MODIFICATIONS SUCH AS DNA METHYLATION AND HISTONE ACETYLATION REGULATE THE EXPRESSION OF GENES AND ALTERATIONS IN THEIR PATTERNS HAVE BEEN LINKED TO MANY INFLAMMATORY DISEASES. THIS STUDY INVESTIGATED WHETHER EPIGENETIC MODIFYING DRUGS AFFECT THE REGULATION OF NOD1/NOD2 ACTIVITY AND EXPRESSION. DNA METHYLTRANSFERASE INHIBITORS HAVE RECENTLY BEEN USED IN THE TREATMENT OF MYELODYSPLASTIC SYNDROME AND AS COMBINATION THERAPY IN CANCER BUT THE FULL EXTENT OF THEIR EFFECTS HAS NOT BEEN QUANTIFIED.METHODS: PHARMACOLOGICAL INHIBITION OF EPIGENETIC ENZYMES IN A HUMAN MONOCYTIC THP-1 CELL LINE WAS CARRIED OUT AND NOD1/NOD2 EXPRESSION AND PRO-INFLAMMATORY RESPONSES WERE QUANTIFIED.RESULTS: CELLS PRIMED WITH A DNA METHYLTRANSFERASE INHIBITOR (BUT NOT A HISTONE DEACETYLASE [HDAC] INHIBITOR) WERE FOUND TO BE CONSISTENTLY MORE RESPONSIVE TO NOD1/NOD2 STIMULATION AND HAD INCREASED BASAL EXPRESSION.CONCLUSION: THE NOVEL EXPERIMENTATION CARRIED OUT HERE SUGGESTS FOR THE FIRST TIME THAT NOD1/NOD2 RECEPTOR ACTIVITY AND EXPRESSION IN MONOCYTES ARE POSSIBLY REGULATED DIRECTLY BY DNA METHYLATION. 2022 19 3662 33 INDUCTION OF STABLE HUMAN FOXP3(+) TREGS BY A PARASITE-DERIVED TGF-BETA MIMIC. IMMUNE HOMEOSTASIS IN THE INTESTINE IS TIGHTLY CONTROLLED BY FOXP3(+) REGULATORY T CELLS (TREGS), DEFECTS OF WHICH ARE LINKED TO THE DEVELOPMENT OF CHRONIC CONDITIONS, SUCH AS INFLAMMATORY BOWEL DISEASE (IBD). AS A MECHANISM OF IMMUNE EVASION, SEVERAL SPECIES OF INTESTINAL PARASITES BOOST TREG ACTIVITY. THE PARASITE HELIGMOSOMOIDES POLYGYRUS IS KNOWN TO SECRETE A MOLECULE (HP-TGM) THAT MIMICS THE ABILITY OF TGF-BETA TO INDUCE FOXP3 EXPRESSION IN CD4(+) T CELLS. THE STUDY AIMED TO INVESTIGATE WHETHER HP-TGM COULD INDUCE HUMAN FOXP3(+) TREGS AS A POTENTIAL THERAPEUTIC APPROACH FOR INFLAMMATORY DISEASES. CD4(+) T CELLS FROM HEALTHY VOLUNTEERS WERE EXPANDED IN THE PRESENCE OF HP-TGM OR TGF-BETA. TREG INDUCTION WAS MEASURED BY FLOW CYTOMETRIC DETECTION OF FOXP3 AND OTHER TREG MARKERS, SUCH AS CD25 AND CTLA-4. EPIGENETIC CHANGES WERE DETECTED USING CHIP-SEQ AND PYROSEQUENCING OF FOXP3. TREG PHENOTYPE STABILITY WAS ASSESSED FOLLOWING INFLAMMATORY CYTOKINE CHALLENGE AND TREG FUNCTION WAS EVALUATED BY CELLULAR CO-CULTURE SUPPRESSION ASSAYS AND CYTOMETRIC BEAD ARRAYS FOR SECRETED CYTOKINES. HP-TGM EFFICIENTLY INDUCED FOXP3 EXPRESSION (> 60%), IN ADDITION TO CD25 AND CTLA-4, AND CAUSED EPIGENETIC MODIFICATION OF THE FOXP3 LOCUS TO A GREATER EXTENT THAN TGF-BETA. HP-TGM-INDUCED TREGS HAD SUPERIOR SUPPRESSIVE FUNCTION COMPARED WITH TGF-BETA-INDUCED TREGS, AND RETAINED THEIR PHENOTYPE FOLLOWING EXPOSURE TO INFLAMMATORY CYTOKINES. FURTHERMORE, HP-TGM INDUCED A TREG-LIKE PHENOTYPE IN IN VIVO DIFFERENTIATED TH1 AND TH17 CELLS, INDICATING ITS POTENTIAL TO RE-PROGRAM MEMORY CELLS TO ENHANCE IMMUNE TOLERANCE. THESE DATA INDICATE HP-TGM HAS POTENTIAL TO BE USED TO GENERATE STABLE HUMAN FOXP3(+) TREGS TO TREAT IBD AND OTHER INFLAMMATORY DISEASES. 2021 20 827 37 CHARACTERIZATION OF LONG NON-CODING RNAS IN SYSTEMIC SCLEROSIS MONOCYTES: A POTENTIAL ROLE FOR PSMB8-AS1 IN ALTERED CYTOKINE SECRETION. SYSTEMIC SCLEROSIS (SSC) IS A CHRONIC AUTOIMMUNE DISEASE MAINLY AFFECTING THE CONNECTIVE TISSUE. IN SSC PATIENTS, MONOCYTES ARE INCREASED IN CIRCULATION, INFILTRATE AFFECTED TISSUES, AND SHOW A PRO-INFLAMMATORY ACTIVATION STATUS, INCLUDING THE SO-CALLED INTERFERON (IFN) SIGNATURE. WE PREVIOUSLY DEMONSTRATED THAT THE DYSREGULATION OF THE IFN RESPONSE IN SSC MONOCYTES IS SUSTAINED BY ALTERED EPIGENETIC FACTORS AS WELL AS BY UPREGULATION OF THE LONG NON-CODING RNA (LNCRNA) NRIR. CONSIDERING THE ENORMOUSLY DIVERSE MOLECULAR FUNCTIONS OF LNCRNAS IN IMMUNE REGULATION, THE PRESENT STUDY INVESTIGATED THE GENOME-WIDE PROFILE OF LNCRNAS IN SSC MONOCYTES, WITH THE AIM TO FURTHER UNRAVEL THEIR POSSIBLE ROLE IN MONOCYTE DYSREGULATION AND DISEASE PATHOGENESIS. TRANSCRIPTOMIC DATA FROM TWO INDEPENDENT COHORTS OF SSC PATIENTS IDENTIFIED 886 LNCRNAS WITH AN ALTERED EXPRESSION IN SSC MONOCYTES. DIFFERENTIALLY EXPRESSED LNCRNAS WERE CORRELATED WITH NEIGHBORING PROTEIN CODING GENES IMPLICATED IN THE REGULATION OF IFN RESPONSES AND APOPTOTIC SIGNALING IN SSC MONOCYTES. IN PARALLEL, GENE CO-EXPRESSION NETWORK ANALYSIS IDENTIFIED THE LNCRNA PSMB8-AS1 AS A TOP-RANKING HUB GENE IN CO-EXPRESSION MODULES IMPLICATED IN CELL ACTIVATION AND RESPONSE TO VIRAL AND EXTERNAL STIMULI. FUNCTIONAL CHARACTERIZATION OF PSMB8-AS1 IN MONOCYTES DEMONSTRATED THAT THIS LNCRNA IS INVOLVED IN THE SECRETION OF IL-6 AND TNFALPHA, TWO PIVOTAL PRO-INFLAMMATORY CYTOKINES ALTERED IN THE CIRCULATION OF SSC PATIENTS AND ASSOCIATED WITH FIBROSIS AND DISEASE SEVERITY. COLLECTIVELY, OUR DATA SHOWED THAT LNCRNAS ARE LINKED TO MONOCYTE DYSREGULATION IN SSC, AND HIGHLIGHT THEIR POTENTIAL CONTRIBUTION TO DISEASE PATHOGENESIS. 2021