1 4041 148 MACROPHAGE PLASTICITY IN DUCHENNE MUSCULAR DYSTROPHY: A NEXUS OF PATHOLOGICAL REMODELLING WITH THERAPEUTIC IMPLICATIONS. DUCHENNE MUSCULAR DYSTROPHY (DMD) IS CHARACTERIZED BY CHRONIC SKELETAL MUSCLE NECROSIS, LEADING TO MUSCLE REGENERATION FAILURE AND FIBROSIS. ALTHOUGH MACROPHAGES (MPS) ARE NORMALLY ESSENTIAL FOR MUSCLE REGENERATION, DYSREGULATED MP FUNCTION PROMOTES PATHOLOGICAL MUSCLE REMODELLING. INFILTRATING MPS CAN BE PREDOMINANTLY PRO-INFLAMMATORY (M1 BIASED), ANTI-INFLAMMATORY (M2 BIASED) OR OF A MIXED PHENOTYPE AND CAN ORIGINATE FROM THE ADULT BONE MARROW (MONOCYTE DEPENDENT) OR EMBRYONIC PRECURSORS (MONOCYTE INDEPENDENT). IN MDX MICE (GENETIC MODEL OF DMD) LACKING EITHER TOLL-LIKE RECEPTOR (TLR) 2 OR TLR4, IT IS FOUND THAT MP INFILTRATION OF DYSTROPHIC MUSCLE IS SIGNIFICANTLY REDUCED AND THAT THE MP PHENOTYPE IS SHIFTED TOWARD A MORE ANTI-INFLAMMATORY PROFILE. THIS IS ACCOMPANIED BY SIGNIFICANT IMPROVEMENTS IN MUSCLE HISTOLOGY AND FORCE PRODUCTION. LACK OF THE CHEMOKINE RECEPTOR CCR2, WHICH IMPEDES MONOCYTE RELEASE FROM THE BONE MARROW, LEADS TO SIMILAR BENEFICIAL EFFECTS IN MDX MICE. EVIDENCE WAS ALSO FOUND FOR TLR4-REGULATED INDUCTION OF TRAINED INNATE IMMUNITY IN MPS CULTURED FROM THE BONE MARROW OF MDX MICE BEFORE THEIR ENTRY INTO THE MUSCLE. THESE MPS EXHIBIT EPIGENETIC AND METABOLIC ALTERATIONS, ACCOMPANIED BY NON-SPECIFIC HYPER-RESPONSIVENESS TO MULTIPLE STIMULI, WHICH IS MANIFESTED BY POTENTIATED UPREGULATION OF BOTH PRO- AND ANTI-INFLAMMATORY GENES. IN SUMMARY, EXAGGERATED RECRUITMENT OF MONOCYTE-DERIVED MPS AND SIGNS OF TRAINED INNATE IMMUNITY AT THE LEVEL OF THE BONE MARROW ARE FEATURES OF THE IMMUNOPHENOTYPE ASSOCIATED WITH DYSTROPHIC MUSCLE DISEASE. THESE PHENOMENA ARE REGULATED BY TOLL-LIKE RECEPTORS THAT BIND ENDOGENOUS DAMAGE-ASSOCIATED MOLECULAR PATTERN (DAMP) MOLECULES, SUGGESTING THAT DAMP RELEASE FROM DYSTROPHIC MUSCLES MODULATES MP PLASTICITY AT THE BONE MARROW LEVEL THROUGH TOLL-LIKE RECEPTOR-DRIVEN MECHANISMS. 2022 2 6495 53 TRAINED IMMUNITY AS A POTENTIAL TARGET FOR THERAPEUTIC IMMUNOMODULATION IN DUCHENNE MUSCULAR DYSTROPHY. DYSREGULATED INFLAMMATION INVOLVING INNATE IMMUNE CELLS, PARTICULARLY OF THE MONOCYTE/MACROPHAGE LINEAGE, IS A KEY CONTRIBUTOR TO THE PATHOGENESIS OF DUCHENNE MUSCULAR DYSTROPHY (DMD). TRAINED IMMUNITY IS AN EVOLUTIONARILY ANCIENT PROTECTIVE MECHANISM AGAINST INFECTION, IN WHICH EPIGENETIC AND METABOLIC ALTERATIONS CONFER NON-SPECIFIC HYPERRESPONSIVENESS OF INNATE IMMUNE CELLS TO VARIOUS STIMULI. RECENT WORK IN AN ANIMAL MODEL OF DMD (MDX MICE) HAS SHOWN THAT MACROPHAGES EXHIBIT CARDINAL FEATURES OF TRAINED IMMUNITY, INCLUDING THE PRESENCE OF INNATE IMMUNE SYSTEM "MEMORY". THE LATTER IS REFLECTED BY EPIGENETIC CHANGES AND DURABLE TRANSMISSIBILITY OF THE TRAINED PHENOTYPE TO HEALTHY NON-DYSTROPHIC MICE BY BONE MARROW TRANSPLANTATION. MECHANISTICALLY, IT IS SUGGESTED THAT A TOLL-LIKE RECEPTOR (TLR) 4-REGULATED, MEMORY-LIKE CAPACITY OF INNATE IMMUNITY IS INDUCED AT THE LEVEL OF THE BONE MARROW BY FACTORS RELEASED FROM THE DAMAGED MUSCLES, LEADING TO EXAGGERATED UPREGULATION OF BOTH PRO- AND ANTI-INFLAMMATORY GENES. HERE WE PROPOSE A CONCEPTUAL FRAMEWORK FOR THE INVOLVEMENT OF TRAINED IMMUNITY IN DMD PATHOGENESIS AND ITS POTENTIAL TO SERVE AS A NEW THERAPEUTIC TARGET. 2023 3 1356 22 DEVELOPMENT OF A DIETARY METHYL DONOR FOOD FREQUENCY QUESTIONNAIRE TO ASSESS FOLATE AND VITAMIN B(12) STATUS IN CHILDREN WITH CHRONIC HEPATITIS B VIRUS INFECTION. OBJECTIVE: TO DEVELOP A DIETARY METHYL DONOR FOOD FREQUENCY QUESTIONNAIRE (DMD-FFQ) THAT IS VALIDATED IN A COHORT OF US CHILDREN AND TO DETERMINE WHETHER THE CONSUMPTION OF FOLATE AND VITAMIN B(12), PRINCIPAL DMDS, CORRELATES WITH HBV DNA LEVELS AND ITS METHYLATION DENSITY. STUDY DESIGN: WE DEVELOPED A SEMIQUANTITATIVE DMD-FFQ TO ESTIMATE INTAKE OF FOLATE AND VITAMIN B(12) AND VALIDATED THIS INSTRUMENT AGAINST A 24-HOUR DIETARY RECALL AND BIOMARKERS-RED BLOOD CELL FOLATE, SERUM VITAMIN B(12), AND HOMOCYSTEINE-IN 35 CHILDREN WITH CHRONIC HBV INFECTION WITHOUT OTHER MEDICAL COMORBIDITIES. ESTIMATES OF DMD, AS WELL AS THE SERUM BIOMARKERS, WERE CORRELATED WITH THE METHYLATION DENSITY OF HBV CPG ISLAND 2 AND HBV DNA LEVELS. RESULTS: FOLATE PER KILOGRAM OF BODY WEIGHT BY THE DMD-FFQ CORRELATED POSITIVELY WITH 24-HOUR RECALL (R = 0.60; P < .001) AND RED BLOOD CELL FOLATE (R = 0.40; P = .02), AND NEGATIVELY WITH HOMOCYSTEINE (R = -0.54; P < .001). VITAMIN B(12) PER KILOGRAM BY DMD-FFQ ALSO CORRELATED POSITIVELY WITH 24-HOUR RECALL (R = 0.57; P < .001) AND SERUM VITAMIN B(12) (R = 0.36, P = .04), AND NEGATIVELY WITH HOMOCYSTEINE (R = -0.44; P = .008). NEITHER DMD INTAKE (FROM DMD-FFQ OR 24-HOUR RECALL) NOR SERUM BIOMARKERS CORRELATED WITH HBV DNA LEVELS OR ITS METHYLATION DENSITY. CONCLUSIONS: OUR DMD-FFQ CORRELATES WELL WITH A 24-HOUR RECALL AND CIRCULATING BIOMARKERS. ALTHOUGH LITTLE EVIDENCE EXISTED THAT CONSUMPTION OF THESE MICRONUTRIENTS CORRELATED WITH HBV REPLICATION, THIS TOOL COULD PROVE USEFUL FOR INVESTIGATING EPIGENETIC MODIFICATION BY DIET FOR SEVERAL PEDIATRIC DISEASES. 2018 4 3307 30 HIGH-RESOLUTION GENOMIC PROFILING OF LIVER CANCER LINKS ETIOLOGY WITH MUTATION AND EPIGENETIC SIGNATURES. BACKGROUND & AIMS: HEPATOCELLULAR CARCINOMA (HCC) IS A MODEL OF A DIVERSE SPECTRUM OF CANCERS BECAUSE IT IS INDUCED BY WELL-KNOWN ETIOLOGIES, MAINLY HEPATITIS C VIRUS (HCV) AND HEPATITIS B VIRUS. HERE, WE AIMED TO IDENTIFY HCV-SPECIFIC MUTATIONAL SIGNATURES AND EXPLORED THE LINK BETWEEN THE HCV-RELATED REGIONAL VARIATION IN MUTATIONS RATES AND HCV-INDUCED ALTERATIONS IN GENOME-WIDE CHROMATIN ORGANIZATION. METHODS: TO IDENTIFY AN HCV-SPECIFIC MUTATIONAL SIGNATURE IN HCC, WE PERFORMED HIGH-RESOLUTION TARGETED SEQUENCING TO DETECT PASSENGER MUTATIONS ON 64 HCC SAMPLES FROM 3 ETIOLOGY GROUPS: HEPATITIS B VIRUS, HCV, OR OTHER. TO EXPLORE THE LINK BETWEEN THE GENOMIC SIGNATURE AND GENOME-WIDE CHROMATIN ORGANIZATION WE PERFORMED CHROMATIN IMMUNOPRECIPITATION SEQUENCING FOR THE TRANSCRIPTIONALLY PERMISSIVE H3K4ME3, H3K9AC, AND SUPPRESSIVE H3K9ME3 MODIFICATIONS AFTER HCV INFECTION. RESULTS: REGIONAL VARIATION IN MUTATION RATE ANALYSIS SHOWED SIGNIFICANT ETIOLOGY-DEPENDENT REGIONAL MUTATION RATES IN 12 GENES: LRP2, KRT84, TMEM132B, DOCK2, DMD, INADL, JAK2, DNAH6, MTMR9, ATM, SLX4, AND ARSD. WE FOUND AN ENRICHMENT OF C->T TRANSVERSION MUTATIONS IN THE HCV-ASSOCIATED HCC CASES. FURTHERMORE, THESE CASES SHOWED REGIONAL VARIATION IN MUTATION RATES ASSOCIATED WITH GENOMIC INTERVALS IN WHICH HCV INFECTION DICTATED EPIGENETIC ALTERATIONS. THIS SIGNATURE MAY BE RELATED TO THE HCV-INDUCED DECREASED EXPRESSION OF GENES ENCODING KEY ENZYMES IN THE BASE EXCISION REPAIR PATHWAY. CONCLUSIONS: WE IDENTIFIED NOVEL DISTINCT HCV ETIOLOGY-DEPENDENT MUTATION SIGNATURES IN HCC ASSOCIATED WITH HCV-INDUCED ALTERATIONS IN HISTONE MODIFICATION. THIS STUDY PRESENTS A LINK BETWEEN CANCER-CAUSING MUTAGENESIS AND THE INCREASED PREDISPOSITION TO LIVER CANCER IN CHRONIC HCV-INFECTED INDIVIDUALS, AND UNVEILS NOVEL ETIOLOGY-SPECIFIC MECHANISMS LEADING TO HCC AND CANCER IN GENERAL. 2023 5 3937 35 LKB1 SIGNALING IS ALTERED IN SKELETAL MUSCLE OF A DUCHENNE MUSCULAR DYSTROPHY MOUSE MODEL. THE POTENTIAL ROLE OF LIVER KINASE B1 (LKB1) IN THE ALTERED ACTIVATION OF THE MASTER METABOLIC AND EPIGENETIC REGULATOR ADENOSINE MONOPHOSPHATE-ACTIVATED PROTEIN KINASE (AMPK) IN DUCHENNE MUSCULAR DYSTROPHY HAS NOT BEEN INVESTIGATED SO FAR. HENCE, WE ANALYZED BOTH GENE AND PROTEIN LEVELS OF LKB1 AND ITS RELATED TARGETS IN GASTROCNEMIUS MUSCLES OF ADULT C57BL/10 MDX MICE AND D2 MDX MICE, A MODEL WITH A MORE SEVERE DYSTROPHIC PHENOTYPE, AS WELL AS THE SENSITIVITY OF THE LKB1-AMPK PATHWAY TO AMPK ACTIVATORS, SUCH AS CHRONIC EXERCISE. OUR DATA SHOW, FOR THE FIRST TIME, A REDUCTION IN THE LEVELS OF LKB1 AND ACCESSORY PROTEINS, MO25 AND STRADALPHA, IN BOTH MDX STRAINS VERSUS THE RESPECTIVE WILD TYPE, WHICH WAS FURTHER IMPAIRED BY EXERCISE, IN PARALLEL WITH A LACK OF FURTHER PHOSPHORYLATION OF AMPK. THE AMPK-LIKE KINASE SALT-INDUCIBLE KINASE (SIK) AND CLASS II HISTONE DEACETYLASES, ALONG WITH EXPRESSION OF THE HDAC TARGET GENE MEF2C, WERE ALSO ALTERED, SUPPORTING AN IMPAIRMENT OF LKB1-SIK-CLASS II HISTONE DEACETYLASE SIGNALING. OUR RESULTS DEMONSTRATE THAT LKB1 MAY BE INVOLVED IN DYSTROPHIC PROGRESSION, PAVING THE WAY FOR FUTURE PRECLINICAL STUDIES. 2023 6 5327 30 PULSED GLUCOCORTICOIDS ENHANCE DYSTROPHIC MUSCLE PERFORMANCE THROUGH EPIGENETIC-METABOLIC REPROGRAMMING. IN HUMANS, CHRONIC GLUCOCORTICOID USE IS ASSOCIATED WITH SIDE EFFECTS LIKE MUSCLE WASTING, OBESITY, AND METABOLIC SYNDROME. INTERMITTENT STEROID DOSING HAS BEEN PROPOSED IN DUCHENNE MUSCULAR DYSTROPHY PATIENTS TO MITIGATE THE SIDE EFFECTS SEEN WITH DAILY STEROID INTAKE. WE EVALUATED BIOMARKERS FROM DUCHENNE MUSCULAR DYSTROPHY PATIENTS, FINDING THAT, COMPARED WITH CHRONIC DAILY STEROID USE, WEEKEND STEROID USE WAS ASSOCIATED WITH REDUCED SERUM INSULIN, FREE FATTY ACIDS, AND BRANCHED CHAIN AMINO ACIDS, AS WELL AS REDUCTION IN FAT MASS DESPITE HAVING SIMILAR BMIS. WE REASONED THAT INTERMITTENT PREDNISONE ADMINISTRATION IN DYSTROPHIC MICE WOULD ALTER MUSCLE EPIGENOMIC SIGNATURES, AND WE IDENTIFIED THE COORDINATED ACTION OF THE GLUCOCORTICOID RECEPTOR, KLF15 AND MEF2C AS MEDIATORS OF A GENE EXPRESSION PROGRAM DRIVING METABOLIC REPROGRAMMING AND ENHANCED NUTRIENT UTILIZATION. MUSCLE LACKING KLF15 FAILED TO RESPOND TO INTERMITTENT STEROIDS. FURTHERMORE, COADMINISTRATION OF THE HISTONE ACETYLTRANSFERASE INHIBITOR ANACARDIC ACID WITH STEROIDS IN MDX MICE ELIMINATED STEROID-SPECIFIC EPIGENETIC MARKS AND ABROGATED THE STEROID RESPONSE. TOGETHER, THESE FINDINGS INDICATE THAT INTERMITTENT, REPEATED EXPOSURE TO GLUCOCORTICOIDS PROMOTES PERFORMANCE IN DYSTROPHIC MUSCLE THROUGH AN EPIGENETIC PROGRAM THAT ENHANCES NUTRIENT UTILIZATION. 2019 7 3794 34 INTERLEUKIN-33 MEDIATES BOTH IMMUNE-RELATED AND NON-IMMUNE-RELATED INHIBITORY EFFECTS AGAINST HEPATITIS B VIRUS. CHRONIC INFECTION BY HEPATITIS B VIRUS (HBV) IS ASSOCIATED WITH HIGH RISKS OF LIVER FIBROSIS, CIRRHOSIS AND HEPATOCELLULAR CARCINOMA. HBV COVALENTLY CLOSED CIRCULAR DNA (CCCDNA) IN THE NUCLEUS OF INFECTED HEPATOCYTE SERVES AS TRANSCRIPTION TEMPLATE. NEITHER NATURAL RESOLUTION OF ACUTE INFECTION NOR CURRENT TREATMENT OPTIONS FOR CHRONIC INFECTION ARE BELIEVED TO CAUSE CCCDNA CLEARANCE. WE PREVIOUSLY SHOWED THAT INJECTION OF IL-33-EXPRESSING PLASMID FACILITATED CLEARANCE OF INTRAHEPATIC HBV DNA IN A MOUSE MODEL OF HBV PERSISTENCE. IN THIS WORK, HBV-TARGETING THERAPEUTIC EFFECTS OF IL-33 WERE FURTHER EXPLORED. MURINE IL-33 DELIVERED BY RECOMBINANT ADENO-ASSOCIATED VIRUS (AAV-MIL-33) INDUCED CLEARANCE OF BOTH SERUM HBV MARKERS AND INTRAHEPATIC HBV DNA IN TWO MOUSE MODELS OF HBV PERSISTENCE BASED ON REPLICON PLASMID AND RECOMBINANT CCCDNA (RCCCDNA) RESPECTIVELY. CLEARANCE WAS ASSOCIATED WITH SERUM ALT ELEVATIONS AND LIVER INFILTRATIONS BY CD4(+) AND CD8(+) T CELLS, INDICATING IL-33-INDUCED CELLULAR IMMUNE RESPONSES AGAINST HBV-HARBORING CELLS. ADOPTIVE TRANSFER OF SPLENOCYTES FROM AAV-MIL-33-CURED MICE WAS INDEED SUFFICIENT TO ENGENDER SIMILAR CLEARANCE IN RECIPIENT MICE. IN VITRO, INTRACELLULAR, INSTEAD OF EXTRACELLULAR, IL-33 WAS MAINLY RESPONSIBLE FOR REPRESSING VIRAL TRANSCRIPTION, PROTEIN PRODUCTION AND GENOME REPLICATION IN HUH7 CELLS TRANSFECTED WITH HBV REPLICON OR RCCCDNA. IL-33 WAS SHOWN TO BE RECRUITED ONTO RCCCDNA MINICHROMOSOME ACCOMPANIED BY LOSS OF TRANSCRIPTIONAL ACTIVATION EPIGENETIC MARKS. FINALLY, TRANSFECTION OF IL-33 INTO HBV-INFECTED HEPG2/NTCP CELLS RESULTED IN REDUCED TRANSCRIPTION, ANTIGEN EXPRESSION AND GENOME REPLICATION, SUGGESTING REPRESSION OF CANONICAL CCCDNA. THESE DATA DEMONSTRATED DIVERSE INHIBITORY EFFECTS ON HBV AND HBV-INFECTED CELLS MEDIATED BY IL-33, AND SUGGEST IL-33 AS AN INTERESTING THERAPEUTIC CANDIDATE. 2022 8 3264 29 HEPATITIS VIRUS INFECTION AFFECTS DNA METHYLATION IN MICE WITH HUMANIZED LIVERS. BACKGROUND & AIMS: CELLS OF TUMORS ASSOCIATED WITH CHRONIC INFLAMMATION FREQUENTLY HAVE ALTERED PATTERNS OF DNA METHYLATION, INCLUDING HEPATOCELLULAR CARCINOMAS. CHRONIC HEPATITIS HAS ALSO BEEN ASSOCIATED WITH ABERRANT DNA METHYLATION, BUT LITTLE IS KNOWN ABOUT THEIR RELATIONSHIP. METHODS: PYROSEQUENCING WAS USED TO DETERMINE THE METHYLATION STATUS OF CULTURED HUH7.5.1 HEPATOMA CELLS AFTER HEPATITIS C VIRUS (HCV) INFECTION. WE ALSO STUDIED MICE WITH SEVERE COMBINED IMMUNODEFICIENCY CARRYING THE UROKINASE-TYPE PLASMINOGEN ACTIVATOR TRANSGENE CONTROLLED BY AN ALBUMIN PROMOTER (UROKINASE-TYPE PLASMINOGEN ACTIVATOR/SEVERE COMBINED IMMUNODEFICIENT MICE), IN WHICH UP TO 85% OF HEPATOCYTES WERE REPLACED BY HUMAN HEPATOCYTES (CHIMERIC MICE). MICE WERE GIVEN INTRAVENOUS INJECTIONS OF HEPATITIS B VIRUS (HBV) OR HCV, LIVER TISSUES WERE COLLECTED, AND DNA METHYLATION PROFILES WERE DETERMINED AT DIFFERENT TIME POINTS AFTER INFECTION. WE ALSO COMPARED METHYLATION PATTERNS BETWEEN PAIRED SAMPLES OF HEPATOCELLULAR CARCINOMAS AND ADJACENT NONTUMOR LIVER TISSUES FROM PATIENTS. RESULTS: NO REPRODUCIBLE CHANGES IN DNA METHYLATION WERE OBSERVED AFTER INFECTION OF HUH7.5.1 CELLS WITH HCV. LIVERS FROM HBV- AND HCV-INFECTED MICE HAD GENOME-WIDE, TIME-DEPENDENT CHANGES IN DNA METHYLATION, COMPARED WITH UNINFECTED UROKINASE-TYPE PLASMINOGEN ACTIVATOR/SEVERE COMBINED IMMUNODEFICIENT MICE. THERE WERE CHANGES IN 160 +/- 63 GENES IN HBV-INFECTED AND 237 +/- 110 GENES IN HCV-INFECTED MICE. METHYLATION OF 149 COMMON GENES INCREASED IN HBV- AND HCV-INFECTED MICE; METHYLATION OF SOME OF THESE GENES ALSO INCREASED IN HEPATOCELLULAR CARCINOMA SAMPLES FROM PATIENTS COMPARED WITH NONTUMOR TISSUES. EXPRESSION OF IFNG, WHICH IS EXPRESSED BY NATURAL KILLER CELLS, INCREASED SIGNIFICANTLY IN CHIMERIC LIVERS, IN CONCORDANCE WITH INDUCTION OF DNA METHYLATION, AFTER INFECTION WITH HBV OR HCV. INDUCTION OF IFNG WAS REDUCED AFTER ADMINISTRATION OF AN INHIBITOR OF NATURAL KILLER CELL FUNCTION (ANTI-ASIALO GM1). CONCLUSIONS: IN CHIMERIC MICE WITH HUMANIZED LIVERS, INFECTION WITH HBV AND HCV APPEARS TO ACTIVATE A NATURAL KILL CELL-DEPENDENT INNATE IMMUNE RESPONSE. THIS CONTRIBUTES TO THE INDUCTION AND ACCUMULATION OF ABERRANT DNA METHYLATION IN HUMAN HEPATOCYTES. 2014 9 2143 28 EPIGENETIC LANDSCAPE OF HIV-1 INFECTION IN PRIMARY HUMAN MACROPHAGE. HUMAN IMMUNODEFICIENCY VIRUS (HIV)-INFECTED MACROPHAGES ARE LONG-LIVED CELLS THAT SUSTAIN PERSISTENT VIRUS EXPRESSION, WHICH IS BOTH A BARRIER TO VIRAL ERADICATION AND CONTRIBUTOR TO NEUROLOGICAL COMPLICATIONS IN PATIENTS DESPITE ANTIRETROVIRAL THERAPY (ART). TO BETTER UNDERSTAND THE REGULATION OF HIV-1 IN MACROPHAGES, WE COMPARED HIV-INFECTED PRIMARY HUMAN MONOCYTE-DERIVED MACROPHAGES (MDM) TO ACUTELY INFECTED PRIMARY CD4 T CELLS AND JURKAT CELLS LATENTLY INFECTED WITH HIV (JLAT 8.4). HIV GENOMES IN MDM WERE ACTIVELY TRANSCRIBED DESPITE ENRICHMENT WITH HETEROCHROMATIN-ASSOCIATED H3K9ME3 ACROSS THE COMPLETE HIV GENOME IN COMBINATION WITH ELEVATED ACTIVATION MARKS OF H3K9AC AND H3K27AC AT THE LONG TERMINAL REPEAT (LTR). MACROPHAGE PATTERNS CONTRASTED WITH JLAT CELLS, WHICH SHOWED CONVENTIONAL BIVALENT H3K4ME3/H3K27ME3, AND ACUTELY INFECTED CD4 T CELLS, WHICH SHOWED AN INTERMEDIATE EPIGENOTYPE. 5'-METHYLCYTOSINE (5MC) WAS ENRICHED ACROSS THE HIV GENOME IN LATENTLY INFECTED JLAT CELLS, WHILE 5'-HYDROXYMETHYLCYTOSINE (5HMC) WAS ENRICHED IN CD4 CELLS AND MDMS. HIV INFECTION INDUCED MULTINUCLEATION OF MDMS ALONG WITH DNA DAMAGE-ASSOCIATED P53 PHOSPHORYLATION, AS WELL AS LOSS OF TET2 AND THE NUCLEAR REDISTRIBUTION OF 5-HYDOXYMETHYLATION. TAKEN TOGETHER, OUR FINDINGS SUGGEST THAT HIV INDUCES A UNIQUE MACROPHAGE NUCLEAR AND TRANSCRIPTIONAL PROFILE, AND VIRAL GENOMES ARE MAINTAINED IN A NONCANONICAL BIVALENT EPIGENETIC STATE. IMPORTANCE MACROPHAGES SERVE AS A RESERVOIR FOR LONG-TERM PERSISTENCE AND CHRONIC PRODUCTION OF HIV. WE FOUND AN ATYPICAL EPIGENETIC CONTROL OF HIV IN MACROPHAGES MARKED BY HETEROCHROMATIC H3K9ME3 DESPITE ACTIVE VIRAL TRANSCRIPTION. HIV INFECTION INDUCED CHANGES IN MACROPHAGE NUCLEAR MORPHOLOGY AND EPIGENETIC REGULATORY FACTORS. THESE FINDINGS MAY IDENTIFY NEW MECHANISMS TO CONTROL CHRONIC HIV EXPRESSION IN INFECTED MACROPHAGES. 2022 10 4831 37 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 11 2185 27 EPIGENETIC MECHANISMS UNDERLYING HIV-INFECTION INDUCED SUSCEPTIBILITY OF CD4+ T CELLS TO ENHANCED ACTIVATION-INDUCED FASL EXPRESSION AND CELL DEATH. BACKGROUND: CHRONIC IMMUNE ACTIVATION AND CD4 T CELL DEPLETION ARE SIGNIFICANT PATHOGENIC FEATURES OF HIV INFECTION. EXPRESSION OF FAS LIGAND (FASL), A KEY MEDIATOR OF ACTIVATION-INDUCED CELL DEATH IN T CELLS, IS ELEVATED IN PEOPLE LIVING WITH HIV-1 INFECTION (PLWH). HOWEVER, THE EPIGENETIC MECHANISMS UNDERLYING THE ENHANCED INDUCTION OF FASL EXPRESSION IN CD4 T LYMPHOCYTES IN PLWH ARE NOT COMPLETELY ELUCIDATED. HENCE, THE CURRENT WORK EXAMINED THE EFFECT OF HIV INFECTION ON FASL PROMOTER-ASSOCIATED HISTONE MODIFICATIONS AND TRANSCRIPTIONAL REGULATION IN CD4 T LYMPHOCYTES IN PLWH. METHOD: FLOW CYTOMETRIC ANALYSIS WAS PERFORMED TO EXAMINE THE FAS-FASL EXPRESSION ON TOTAL CD4 T CELLS AND NAIVE/MEMORY CD4 T CELL SUBSETS. EPIGENETIC FASL PROMOTER HISTONE MODIFICATIONS WERE INVESTIGATED BY CHROMATIN IMMUNOPRECIPITATION-QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION ANALYSIS USING FRESHLY ISOLATED TOTAL CD4 T LYMPHOCYTES FROM HIV-1 INFECTED AND NONINFECTED INDIVIDUALS. RESULTS: ALL NAIVE/MEMORY CD4 T CELL SUBSETS FROM PLWH SHOWED MARKEDLY GREATER FREQUENCY OF FASL EXPRESSION. NOTABLY, EXAMINATION OF FUNCTIONAL OUTCOME OF FASL/FAS CO-EXPRESSION DEMONSTRATED THE PREFERENTIAL SUSCEPTIBILITY OF TCM AND TEM SUBSETS TO ACTIVATION-INDUCED APOPTOSIS. IMPORTANTLY, THESE CD4 T CELLS COLLECTIVELY DEMONSTRATED A DISTINCT FASL PROMOTER HISTONE PROFILE INVOLVING A COORDINATED CROSS-TALK BETWEEN HISTONE H3 MODIFICATIONS LEADING TO ENHANCED FASL GENE EXPRESSION. SPECIFICALLY, LEVELS OF TRANSCRIPTIONALLY PERMISSIVE HISTONE H3K4-TRIMETHYLATION (H3K4ME3) AND HISTONE H3K9-ACETYLATION (H3K9AC) WERE INCREASED, WITH A CONCOMITANT DECREASE IN THE REPRESSIVE H3K9-TRIMETHYLATION (H3K9ME3). CONCLUSION: THE PRESENT WORK DEMONSTRATES THAT EPIGENETIC MECHANISMS INVOLVING PROMOTER-HISTONE MODIFICATIONS REGULATE TRANSCRIPTIONAL COMPETENCE AND FASL EXPRESSION IN CD4 T CELLS FROM PLWH AND RENDER THEM SUSCEPTIBLE TO ACTIVATION-INDUCED CELL DEATH. 2021 12 2476 35 EPIGENETIC UPREGULATION OF CCL2 AND CCL3 VIA HISTONE MODIFICATIONS IN INFILTRATING MACROPHAGES AFTER PERIPHERAL NERVE INJURY. TO GAIN INSIGHT INTO THE EPIGENETIC REGULATION OF CC-CHEMOKINE LIGAND (CCL) 2 AND CCL3, KEY PLAYERS IN THE PERIPHERAL SENSITIZATION LEADING TO NEUROPATHIC PAIN, WE EXAMINED THE RELATIONSHIP BETWEEN HISTONE H3 MODIFICATION AND THE UPREGULATION OF THESE MOLECULES USING A MOUSE MODEL OF NEUROPATHIC PAIN AFTER PARTIAL SCIATIC NERVE LIGATION (PSL). WE FOUND THAT CIRCUITING BONE MARROW (BM)-DERIVED MACROPHAGES INFILTRATED INTO THE INJURED SCIATIC NERVE (SCN) USING ENHANCED GREEN FLUORESCENT PROTEIN CHIMERIC MICE. THE MRNA LEVELS OF CCL2, CCL3 AND THEIR RECEPTORS (CCR2 AND CCR1/CCR5, RESPECTIVELY) WERE INCREASED IN THE INJURED SCN. CHROMATIN IMMUNOPRECIPITATION ASSAY REVEALED THAT LEVELS OF LYSINE 9-ACETYLATED HISTONE H3 (H3K9AC) AND LYSINE 4-TRIMETHYLATED H3 (H3K4ME(3)) IN THE PROMOTER REGIONS OF THE CCL2 AND CCL3 GENES WERE INCREASED IN THE INJURED SCN AFTER PSL, INDICATING THE ENHANCEMENT OF GENE EXPRESSION. IMMUNOREACTIVITY FOR H3K9AC AND H3K4ME(3) WAS LOCALIZED IN THE NUCLEI OF INFILTRATING BM-DERIVED CELLS AND CCL-EXPRESSING CELLS IN THE INJURED SCN. WE OBSERVED H3K9AC AND H3K4ME(3) MAINLY IN THE NUCLEI OF RECRUITED MACROPHAGES ON DAY 7 AFTER PSL. FURTHERMORE, UPREGULATION OF CCLS AND CCRS WERE SUPPRESSED BY HISTONE ACETYLTRANSFERASE INHIBITOR, ANACARDIC ACID. TAKEN TOGETHER, OUR FINDINGS DEMONSTRATE THAT CCL2 AND CCL3 ARE UPREGULATED IN THE INJURED PERIPHERAL NERVE THROUGH EPIGENETIC HISTONE MODIFICATION IN INFILTRATING IMMUNE CELLS SUCH AS MACROPHAGES. THESE CHEMOKINE CASCADES MAY SUBSEQUENTLY ELICIT CHRONIC NEUROINFLAMMATION FOLLOWING NERVE INJURY. 2013 13 2312 30 EPIGENETIC REGULATION OF DENDRITIC CELL-DERIVED INTERLEUKIN-12 FACILITATES IMMUNOSUPPRESSION AFTER A SEVERE INNATE IMMUNE RESPONSE. PATIENTS WHO SURVIVE SEPSIS HAVE SIGNIFICANT DEFICIENCIES IN THEIR IMMUNE RESPONSES CAUSED BY POORLY UNDERSTOOD MECHANISMS. WE HAVE EXPLORED THIS PHENOMENON BY STUDYING DENDRITIC CELLS (DCS) RECOVERED FROM ANIMALS SURVIVING SEVERE PERITONITIS-INDUCED SEPSIS, USING THE WELL-ESTABLISHED CECAL LIGATION AND PUNCTURE (CLP) MODEL. IMMEDIATELY AFTER THE INITIATION OF SEPSIS THERE IS A DEPLETION IN DCS FROM THE LUNG AND SPLEEN, WHICH IS FOLLOWED BY REPOPULATION OF THESE CELLS BACK TO THE RESPECTIVE ORGANS. DCS RECOVERED FROM SURVIVING ANIMALS EXHIBITED A SIGNIFICANT AND CHRONIC SUPPRESSION OF INTERLEUKIN-12 (IL-12), A KEY HOST DEFENSE CYTOKINE. THE SUPPRESSION OF DC-DERIVED IL-12 PERSISTED FOR AT LEAST 6 WEEKS AFTER CLP AND WAS NOT DUE TO IMMUNOREGULATORY CYTOKINES, SUCH AS IL-10. USING CHROMATIN IMMUNOPRECIPITATION (CHIP) TECHNIQUES, WE HAVE SHOWN THAT THE DEFICIENCY IN DC-DERIVED IL-12 WAS DUE TO EPIGENETIC ALTERATIONS. SPECIFICALLY, IL-12 EXPRESSION WAS REGULATED BY STABLE RECIPROCAL CHANGES IN HISTONE H3 LYSINE-4 TRIMETHYLATION (H3K4ME3) AND HISTONE H3 LYSINE-27 DIMETHYLATION (H3K27ME2), AS WELL AS CHANGES IN COGNATE HISTONE METHYLTRANSFERASE (HMT) COMPLEXES ON THE IL12P35 AND IL12P40 PROMOTERS. THESE DATA IMPLICATE HISTONE MODIFICATION ENZYMES IN SUPPRESSING DC-DERIVED IL-12, WHICH MAY PROVIDE ONE OF THE MECHANISMS OF LONG-TERM IMMUNOSUPPRESSION SUBSEQUENT TO THE SEPTIC RESPONSE. 2008 14 3619 27 IN VIVO ACUTE ON CHRONIC ETHANOL EFFECTS IN LIVER: A MOUSE MODEL EXHIBITING EXACERBATED INJURY, ALTERED METABOLIC AND EPIGENETIC RESPONSES. CHRONIC ALCOHOLICS WHO ALSO BINGE DRINK (I.E., ACUTE ON CHRONIC) ARE PRONE TO AN EXACERBATED LIVER INJURY BUT ITS MECHANISM IS NOT UNDERSTOOD. WE THEREFORE INVESTIGATED THE IN VIVO EFFECTS OF CHRONIC AND BINGE ETHANOL INGESTION AND COMPARED TO CHRONIC ETHANOL FOLLOWED BY THREE REPEAT BINGE ETHANOL ON THE LIVER OF MALE C57/BL6 MICE FED ETHANOL IN LIQUID DIET (4%) FOR FOUR WEEKS FOLLOWED BY BINGE ETHANOL (INTRAGASTRIC ADMINISTRATION, 3.5 G/KG BODY WEIGHT, THREE DOSES, 12H APART). CHRONIC FOLLOWED BY BINGE ETHANOL EXACERBATED FAT ACCUMULATION, NECROSIS, DECREASE IN HEPATIC SAM AND SAM:SAH RATIO, INCREASE IN ADENOSINE LEVELS, AND ELEVATED CYP2E1 LEVELS. HISTONE H3 LYSINE ACETYLATION (H3ACK9), DUALLY MODIFIED PHOSPHOACETYLATED HISTONE H3 (H3ACK9/PS10), AND PHOSPHORYLATED H2AX INCREASED AFTER BINGE WHEREAS PHOSPHORYLATION OF HISTONE H3 SER 10 (H3S10) AND H3 SER 28 (H3S28) INCREASED AFTER CHRONIC ETHANOL-BINGE. HISTONE H3 LYSINE 4 AND 9 DIMETHYLATION INCREASED WITH A MARKED DIMETHYLATION IN H3K9 IN CHRONIC ETHANOL BINGE GROUP. TRIMETHYLATED HISTONE H3 LEVELS DID NOT CHANGE. NUCLEAR LEVELS OF HISTONE ACETYL TRANSFERASE GCN5 AND HISTONE DEACETYLASE HDAC3 WERE ELEVATED WHEREAS PHOSPHO-CREB DECREASED IN A DISTINCTIVE MANNER. TAKEN TOGETHER, ACUTE ON CHRONIC ETHANOL INGESTION CAUSED AMPLIFICATION OF LIVER INJURY AND ELICITED CHARACTERISTIC PROFILES OF HISTONE MODIFICATIONS, METABOLIC ALTERATIONS, AND CHANGES IN NUCLEAR PROTEIN LEVELS. THESE FINDINGS DEMONSTRATE THAT CHRONIC ETHANOL EXPOSURE RENDERS LIVER MORE SUSCEPTIBLE TO REPEAT ACUTE/BINGE ETHANOL INDUCED ACCELERATION OF ALCOHOLIC LIVER DISEASE. 2015 15 3246 19 HEPATITIS B VIRUS (HBV) INDUCES THE EXPRESSION OF INTERLEUKIN-8 THAT IN TURN REDUCES HBV SENSITIVITY TO INTERFERON-ALPHA. HIGH LEVELS OF SERUM INTERLEUKIN-8 (IL-8) HAVE BEEN DETECTED IN CHRONIC HEPATITIS B (CHB) PATIENTS DURING EPISODES OF HEPATITIS FLARES. WE INVESTIGATED WHETHER HEPATITIS B VIRUS (HBV) MAY DIRECTLY INDUCE IL-8 PRODUCTION AND WHETHER IL-8 MAY ANTAGONIZE INTERFERON-ALPHA (IFN-ALPHA) ANTIVIRAL ACTIVITY AGAINST HBV. WE SHOWED THAT CHB PATIENTS HAD SIGNIFICANTLY HIGHER IL-8 LEVELS BOTH IN SERUM AND IN LIVER TISSUE THAN CONTROLS. IN HBV-REPLICATING HEPG2 CELLS, IL-8 TRANSCRIPTION WAS SIGNIFICANTLY ACTIVATED. AP-1, C/EBP AND NF-KB TRANSCRIPTION FACTORS WERE CONCURRENTLY NECESSARY FOR MAXIMUM IL-8 INDUCTION. MOREOVER, HBX VIRAL PROTEIN WAS RECRUITED ONTO THE IL-8 PROMOTER AND THIS WAS PARALLELED BY IL8-BOUND HISTONE HYPERACETYLATION AND BY ACTIVE RECRUITMENT OF TRANSCRIPTIONAL COACTIVATORS. INHIBITION OF IL-8 INCREASES THE ANTIVIRAL ACTIVITY OF IFN-ALPHA AGAINST HBV. OUR RESULTS INDICATE THAT HBV ACTIVATES IL-8 GENE EXPRESSION BY TARGETING THE EPIGENETIC REGULATION OF THE IL-8 PROMOTER AND THAT IL-8 MAY CONTRIBUTE TO REDUCE HBV SENSITIVITY TO IFN-ALPHA. 2013 16 3760 27 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 17 2120 24 EPIGENETIC HISTONE MODIFICATIONS IN A CLINICALLY RELEVANT RAT MODEL OF CHRONIC ETHANOL-BINGE-MEDIATED LIVER INJURY. PURPOSE: ETHANOL BINGE AUGMENTS LIVER INJURY AFTER CHRONIC ETHANOL CONSUMPTION IN HUMANS, BUT THE MECHANISM BEHIND THE ENHANCED LIVER INJURY BY ETHANOL BINGE IS NOT KNOWN. IN THIS STUDY WE USED A CLINICALLY RELEVANT RAT MODEL IN WHICH LIVER INJURY IS AMPLIFIED BY BINGE AFTER CHRONIC ETHANOL TREATMENT AND INVESTIGATED THE IMPORTANCE OF HISTONE MODIFICATIONS. METHODS: EIGHT-WEEK-OLD SPRAGUE-DAWLEY RATS WERE FED ETHANOL IN A LIQUID DIET FOR 4 WEEKS. CONTROL RATS WERE FED AN ISOCALORIC LIQUID DIET. THIS WAS FOLLOWED BY THREE BINGE ADMINISTRATIONS OF ETHANOL (INTRAGASTRIC 5 G/KG BODY WEIGHT, 12 H APART). IN THE CONTROL, ETHANOL WAS REPLACED BY WATER. FOUR HOURS AFTER THE LAST BINGE ADMINISTRATION, LIVER SAMPLES WERE ANALYZED FOR HISTONE MODIFICATIONS AND PARAMETERS OF LIVER INJURY. RESULTS: CHRONIC ETHANOL ADMINISTRATION ALONE CAUSED AN INCREASE IN HISTONE H3 SER10 AND SER28 (H3S10 OR S28) PHOSPHORYLATION, AND BINGE ETHANOL REDUCED THEIR LEVELS. LEVELS OF DUALLY MODIFIED PHOSPHOACETYLATED HISTONE H3 (H3ACK9/PS10) INCREASED AFTER ACUTE BINGE ETHANOL AND REMAINED SAME AFTER CHRONIC ETHANOL BINGE. IN CONTRAST, HISTONE H3 LYSINE-9 ACETYLATION (H3ACK9) WAS NOT INCREASED AFTER CHRONIC ETHANOL BUT INCREASED SIGNIFICANTLY AFTER ACUTE BINGE AND CHRONIC ETHANOL BINGE. INCREASE IN HISTONE ACETYLATION WAS ACCOMPANIED BY INCREASED PHOSPHO-ERK1/2 IN THE NUCLEAR EXTRACTS. INCREASED ACETYLATION AFTER CHRONIC ETHANOL BINGE WAS ALSO ACCOMPANIED BY INCREASED PROTEIN LEVELS OF GCN5 HISTONE ACETYL TRANSFERASE AND A MODEST INCREASE IN HDAC3 IN THE NUCLEUS. HISTONE LYSINE-9 DIMETHYLATION WAS SIGNIFICANTLY INCREASED AFTER CHRONIC ETHANOL BINGE. CHRONIC ETHANOL BINGE ALSO RESULTED IN A DECREASE IN THE SAM:SAH RATIO WITH A RELATIVE DECREASE OF SAM LEVELS AND A CORRESPONDING INCREASE IN SAH LEVELS. CONCLUSIONS: ETHANOL BINGE AFTER CHRONIC ETHANOL ALTERED THE PROFILE OF SITE-SPECIFIC HISTONE MODIFICATIONS AND MAY UNDERLIE THE MECHANISM OF AUGMENTED LIVER INJURY BY CHRONIC-ETHANOL-BINGE-TREATED RATS. 2014 18 3088 25 GENOMEWIDE HISTONE H3 LYSINE 9 ACETYLATION PROFILING IN CD4+ T CELLS REVEALED ENDOPLASMIC RETICULUM STRESS DEFICIENCY IN PATIENTS WITH ACUTE-ON-CHRONIC LIVER FAILURE. ACUTE-ON-CHRONIC LIVER FAILURE (ACLF) DISPLAYED 'SEPSIS-LIKE' IMMUNE PARALYSIS. LITTLE IS KNOWN ABOUT THE ROLE OF CD4+ T LYMPHOCYTES, THE PRIMARY REGULATOR OF INNATE AND ADOPTED IMMUNE SYSTEM, PLAYED IN ACLF. ACETYLATION OF HISTONE H3 LYSINE 9 (H3K9AC), A KEY EPIGENETIC MODIFICATION, TIGHTLY CONTROLS GENE TRANSCRIPTION. WHETHER AND HOW DOES H3K9AC MODIFICATION REGULATE CD4+ T CELLS IN ACLF REMAINS UNCLEAR. PBMCS WERE ISOLATED FROM PATIENTS WITH ACLF, IMMUNE TOLERANCE OF CHRONIC HEPATITIS B (CHB-T) AND IMMUNE ACTIVE OF CHRONIC HEPATITIS B (CHB-A). THEN, CD4+ T LYMPHOCYTES WERE PURIFIED BY MAGNETIC MICROBEADS, AND THE PURITY WAS CONFIRMED BY FLOW CYTOMETRY. H3K9AC VARIATIONS WERE ANALYSED IN CD4+ T CELLS USING CHROMATIN IMMUNOPRECIPITATION MICROARRAY AND THEN CONFIRMED BY QUANTITATIVE PCR. WHOLE-GENOME H3K9 ACETYLATION ANALYSES WERE CONDUCTED BY BIOINFORMATICS. A TOTAL OF 70 GENES WERE DIFFERENTLY MODIFIED IN H3K9AC BETWEEN CHB-A AND ACLF GROUPS, WHILE 44 GENES WERE DIFFERENTLY MODIFIED IN H3K9AC BETWEEN CHB-T AND ACLF GROUPS. CLUSTERING ALGORITHM ANALYSIS SHOWED PATIENTS WITH ACLF DISPLAYED 'SEPSIS-LIKE' IMMUNE PARALYSIS. FUNCTIONAL ANALYSIS SHOWED ENDOPLASMIC RETICULUM (ER) STRESS, OR DOWNSTREAM PATHWAY-RELATED GENES, SUCH AS BIP, ATF4, PER1, CSNK1D, IRF3, BNIP1, AKT1 AND UBC, WERE DIFFERENTIALLY MODIFIED IN ACLF. WE PROFILED H3K9 ACETYL MODIFICATION IN CD4+ T LYMPHOCYTES FROM HBV-INFECTED PATIENTS WITH THREE DIFFERENT IMMUNE STATES, THAT IS ACLF, IMMUNE TOLERANCE AND IMMUNE ACTIVE PHASES. ACLF DISPLAYED 'SEPSIS-LIKE' IMMUNE PARALYSIS. ER STRESS IN CD4+ T LYMPHOCYTES ATTRIBUTED TO ACLF. THIS STUDY PROVIDES SOME USEFUL CLUES FOR REVEALING THE MECHANISMS UNDERLYING ACLF. 2015 19 6540 35 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 20 744 42 CANNABINOID WIN55,212-2 REPROGRAMS MONOCYTES AND MACROPHAGES TO INHIBIT LPS-INDUCED INFLAMMATION. INTRODUCTION: CHRONIC OR UNCONTROLLED ACTIVATION OF MYELOID CELLS INCLUDING MONOCYTES, MACROPHAGES AND DENDRITIC CELLS (DCS) IS A HALLMARK OF IMMUNE-MEDIATED INFLAMMATORY DISORDERS. THERE IS AN URGENT NEED FOR THE DEVELOPMENT OF NOVEL DRUGS WITH THE CAPACITY TO IMPAIR INNATE IMMUNE CELL OVERACTIVATION UNDER INFLAMMATORY CONDITIONS. COMPELLING EVIDENCE POINTED OUT CANNABINOIDS AS POTENTIAL THERAPEUTIC TOOLS WITH ANTI-INFLAMMATORY AND IMMUNOMODULATORY CAPACITY. WIN55,212-2, A NON-SELECTIVE SYNTHETIC CANNABINOID AGONIST, DISPLAYS PROTECTIVE EFFECTS IN SEVERAL INFLAMMATORY CONDITIONS BY MECHANISMS PARTIALLY DEPENDING ON THE GENERATION OF TOLEROGENIC DCS ABLE TO INDUCE FUNCTIONAL REGULATORY T CELLS (TREGS). HOWEVER, ITS IMMUNOMODULATORY CAPACITY ON OTHER MYELOID CELLS SUCH AS MONOCYTES AND MACROPHAGES REMAINS INCOMPLETELY UNDERSTOOD. METHODS: HUMAN MONOCYTE-DERIVED DCS (HMODCS) WERE DIFFERENTIATED IN THE ABSENCE (CONVENTIONAL HMODCS) OR PRESENCE OF WIN55,212-2 (WIN-HMODCS). CELLS WERE STIMULATED WITH LPS, COCULTURED WITH NAIVE T LYMPHOCYTES AND THEIR CYTOKINE PRODUCTION AND ABILITY TO INDUCE T CELL RESPONSES WERE ANALYSED BY ELISA OR FLOW CYTOMETRY. TO EVALUATE THE EFFECT OF WIN55,212-2 IN MACROPHAGE POLARIZATION, HUMAN AND MURINE MACROPHAGES WERE ACTIVATED WITH LPS OR LPS/IFNGAMMA, IN THE PRESENCE OR ABSENCE OF THE CANNABINOID. CYTOKINE, COSTIMULATORY MOLECULES AND INFLAMMASOME MARKERS WERE ASSAYED. METABOLIC AND CHROMATIN IMMUNOPRECIPITATION ASSAYS WERE ALSO PERFORMED. FINALLY, THE PROTECTIVE CAPACITY OF WIN55,212-2 WAS STUDIED IN VIVO IN BALB/C MICE AFTER INTRAPERITONEAL INJECTION WITH LPS. RESULTS: WE SHOW FOR THE FIRST TIME THAT THE DIFFERENTIATION OF HMODCS IN THE PRESENCE OF WIN55,212-2 GENERATES TOLEROGENIC WIN-HMODCS THAT ARE LESS RESPONSIVE TO LPS STIMULATION AND ABLE TO PRIME TREGS. WIN55,212-2 ALSO IMPAIRS THE PRO-INFLAMMATORY POLARIZATION OF HUMAN MACROPHAGES BY INHIBITING CYTOKINE PRODUCTION, INFLAMMASOME ACTIVATION AND RESCUING MACROPHAGES FROM PYROPTOTIC CELL DEATH. MECHANISTICALLY, WIN55,212-2 INDUCED A METABOLIC AND EPIGENETIC SHIFT IN MACROPHAGES BY DECREASING LPS-INDUCED MTORC1 SIGNALING, COMMITMENT TO GLYCOLYSIS AND ACTIVE HISTONE MARKS IN PRO-INFLAMMATORY CYTOKINE PROMOTERS. WE CONFIRMED THESE DATA IN EX VIVO LPS-STIMULATED PERITONEAL MACROPHAGES (PMPHIS), WHICH WERE ALSO SUPPORTED BY THE IN VIVO ANTI-INFLAMMATORY CAPACITY OF WIN55,212-2 IN A LPS-INDUCED SEPSIS MOUSE MODEL. CONCLUSION: OVERALL, WE SHED LIGHT INTO THE MOLECULAR MECHANISMS BY WHICH CANNABINOIDS EXERT ANTI-INFLAMMATORY PROPERTIES IN MYELOID CELLS, WHICH MIGHT WELL CONTRIBUTE TO THE FUTURE RATIONAL DESIGN OF NOVEL THERAPEUTIC STRATEGIES FOR INFLAMMATORY DISORDERS. 2023