1 3937 118 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 2 4041 35 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 3 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 4 4576 32 MYOGENIC POTENTIAL OF CANINE CRANIOFACIAL SATELLITE CELLS. THE SKELETAL FIBERS HAVE DIFFERENT EMBRYOLOGICAL ORIGIN; THE EXTRAOCULAR AND JAW-CLOSER MUSCLES DEVELOP FROM PRECHORDAL MESODERM WHILE THE LIMB AND TRUNK MUSCLES FROM SOMITES. THESE DIFFERENT ORIGINS CHARACTERIZE ALSO THE ADULT MUSCLE STEM CELLS, KNOWN AS SATELLITE CELLS (SCS) AND RESPONSIBLE FOR THE FIBER GROWTH AND REGENERATION. THE PHYSIOLOGICAL PROPERTIES OF PRESOMITIC SCS AND THEIR EPIGENETICS ARE POORLY STUDIED DESPITE THEIR PECULIAR CHARACTERISTICS TO PRESERVE MUSCLE INTEGRITY DURING CHRONIC MUSCLE DEGENERATION. HERE, WE ISOLATED SCS FROM CANINE SOMITIC [SOMITE-DERIVED MUSCLE (SDM): VASTUS LATERALIS, RECTUS ABDOMINIS, GLUTEUS SUPERFICIALIS, BICEPS FEMORIS, PSOAS] AND PRESOMITIC [PRE-SOMITE-DERIVED MUSCLE (PSDM): LATERAL RECTUS, TEMPORALIS, AND RETRACTOR BULBI] MUSCLES AS MYOGENIC PROGENITOR CELLS FROM YOUNG AND OLD ANIMALS. IN ADDITION, SDM AND PSDM-SCS WERE OBTAINED ALSO FROM GOLDEN RETRIEVERS AFFECTED BY MUSCULAR DYSTROPHY (GRMD). WE CHARACTERIZED THE LIFESPAN, THE MYOGENIC POTENTIAL AND FUNCTIONS, AND OXIDATIVE STRESS OF BOTH SOMITIC AND PRESOMITIC SCS WITH THE AIM TO REVEAL DIFFERENCES WITH AGING AND BETWEEN HEALTHY AND DYSTROPHIC ANIMALS. THE DIFFERENT PROLIFERATION RATE WAS CONSISTENT WITH HIGHER TELOMERASE ACTIVITY IN PSDM-SCS COMPARED TO SDM-SCS, ALTHOUGH RESTRICTED AT EARLY PASSAGES. SDM-SCS EXPRESS EARLY (PAX7, MYOD) AND LATE (MYOSIN HEAVY CHAIN, MYOGENIN) MYOGENIC MARKERS DIFFERENTLY FROM PSDM-SCS RESULTING IN A MORE EFFICIENT AND FASTER CELL DIFFERENTIATION. TAKEN TOGETHER, OUR RESULTS SHOWED THAT PSDM-SCS ELICIT A STRONGER STEM CELL PHENOTYPE COMPARED TO SDM ONES. FINALLY, MYOMIR EXPRESSION PROFILE REVEALS A UNIQUE EPIGENETIC SIGNATURE IN GRMD SCS AND MIR-206, HIGHLY EXPRESSED IN DYSTROPHIC SCS, SEEMS TO PLAY A CRITICAL ROLE IN MUSCLE DEGENERATION. THUS, MIR-206 COULD REPRESENT A POTENTIAL TARGET FOR NOVEL THERAPEUTIC APPROACHES. 2014 5 6495 30 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 6 545 23 ATTENUATED EPIGENETIC SUPPRESSION OF MUSCLE STEM CELL NECROPTOSIS IS REQUIRED FOR EFFICIENT REGENERATION OF DYSTROPHIC MUSCLES. SOMATIC STEM CELLS EXPAND MASSIVELY DURING TISSUE REGENERATION, WHICH MIGHT REQUIRE CONTROL OF CELL FITNESS, ALLOWING ELIMINATION OF NON-COMPETITIVE, POTENTIALLY HARMFUL CELLS. HOW OR IF SUCH CELLS ARE REMOVED TO RESTORE ORGAN FUNCTION IS NOT FULLY UNDERSTOOD. HERE, WE SHOW THAT A SUBSTANTIAL FRACTION OF MUSCLE STEM CELLS (MUSCS) UNDERGO NECROPTOSIS BECAUSE OF EPIGENETIC REWIRING DURING CHRONIC SKELETAL MUSCLE REGENERATION, WHICH IS REQUIRED FOR EFFICIENT REGENERATION OF DYSTROPHIC MUSCLES. INHIBITION OF NECROPTOSIS STRONGLY ENHANCES SUPPRESSION OF MUSC EXPANSION IN A NON-CELL-AUTONOMOUS MANNER. PREVENTION OF NECROPTOSIS IN MUSCS OF HEALTHY MUSCLES IS MEDIATED BY THE CHROMATIN REMODELER CHD4, WHICH DIRECTLY REPRESSES THE NECROPTOTIC EFFECTOR RIPK3, WHILE CHD4-DEPENDENT RIPK3 REPRESSION IS DRAMATICALLY ATTENUATED IN DYSTROPHIC MUSCLES. LOSS OF RIPK3 REPRESSION BY INACTIVATION OF CHD4 CAUSES MASSIVE NECROPTOSIS OF MUSCS, ABOLISHING REGENERATION. OUR STUDY DEMONSTRATES HOW PROGRAMMED CELL DEATH IN MUSCS IS TIGHTLY CONTROLLED TO ACHIEVE OPTIMAL TISSUE REGENERATION. 2020 7 550 19 AUTOIMMUNE POLYENDOCRINOPATHY-CANDIDIASIS-ECTODERMAL DYSTROPHY IN TWO SIBLINGS: SAME MUTATIONS BUT VERY DIFFERENT PHENOTYPES. AUTOIMMUNE POLYENDOCRINOPATHY-CANDIDIASIS-ECTODERMAL DYSTROPHY (APECED), CAUSED BY MUTATIONS IN THE AIRE GENE, IS MAINLY CHARACTERIZED BY THE TRIAD OF HYPOPARATHYROIDISM, PRIMARY ADRENOCORTICAL INSUFFICIENCY AND CHRONIC MUCOCUTANEOUS CANDIDIASIS, BUT CAN INCLUDE MANY OTHER MANIFESTATIONS, WITH NO CURRENTLY CLEAR GENOTYPE-PHENOTYPE CORRELATION. WE PRESENT THE CLINICAL FEATURES OF TWO SIBLINGS, A MALE AND A FEMALE, WITH THE SAME MUTATIONS IN THE AIRE GENE ASSOCIATED WITH TWO VERY DIFFERENT PHENOTYPES. INTERESTINGLY, THE BROTHER RECENTLY EXPERIENCED COVID-19 INFECTION WITH PNEUMONIA, COMPLICATED BY HYPERTENSION, HYPOKALEMIA AND HYPERCALCEMIA. ALTHOUGH APECED IS A MONOGENIC DISEASE, ITS EXPRESSIVENESS CAN BE EXTREMELY DIFFERENT. IN ADDITION TO THE GENETIC BASIS, EPIGENETIC AND ENVIRONMENTAL FACTORS MIGHT INFLUENCE THE PHENOTYPIC EXPRESSION, ALTHOUGH THEIR EXACT ROLE REMAINS TO BE ELUCIDATED. 2021 8 553 15 AUTOINFLAMMATORY RETINOPATHY IN CHRONIC INFANTILE NEUROLOGICAL CUTANEOUS AND ARTICULAR (CINCA) SYNDROME. CHRONIC INFANTILE NEUROLOGICAL CUTANEOUS AND ARTICULAR (CINCA) SYNDROME IS A RARE AUTOSOMAL DOMINANT AUTOINFLAMMATORY DISEASE. WE REPORT THE CASES OF MONOZYGOTIC TWINS WITH CINCA SYNDROME WHOSE PREDOMINANT OCULAR MANIFESTATION WAS INFLAMMATORY ROD-CONE RETINAL DYSTROPHY. ATYPICALLY, THERE WERE SIGNIFICANT DIFFERENCES BETWEEN TWINS IN PHENOTYPE SEVERITY, SUGGESTIVE OF EPIGENETIC DIFFERENCES AND/OR INVOLVEMENT OF ENVIRONMENTAL FACTORS. 2016 9 6817 20 [FACIOSCAPULOHUMERAL MUSCULAR DYSTROPHY: REPORT OF SEVEN PATIENTS]. BACKGROUND: FACIOSCAPULOHUMERAL MUSCULAR DYSTROPHY IS THE THIRD MOST COMMON MUSCULAR DYSTROPHY WITH AN ESTIMATED PREVALENCE OF 1 PER 20.000 AND A NORMAL LIFE EXPECTANCY IN THE MAJORITY OF PATIENTS. HOWEVER, APPROXIMATELY 15% OF PATIENTS BECOME WHEELCHAIR BOUND IN THE COURSE OF THEIR LIFE. IT IS A HEREDITARY AUTOSOMAL DOMINANT DISEASE WITH HIGH (95%) PENETRANCE BY THE AGE OF 20, BUT WITH VARIABLE DEGREE OF PHENOTYPIC EXPRESSION EVEN IN THE SAME FAMILY GROUP. SYMPTOMS FREQUENTLY START IN THE SECOND DECADE OF LIFE, WITH FACIAL AND SCAPULAR WEAKNESS. AIM: TO REPORT THE CLINICAL FEATURES OF SEVEN PATIENTS WITH THE DISEASE, SEEN AT A PUBLIC HOSPITAL. MATERIAL AND METHODS: ANALYSIS OF SEVEN PATIENTS WITH GENETIC STUDY SEEN IN A PUBLIC HOSPITAL IN SANTIAGO. RESULTS: THE AGE OF PATIENTS FLUCTUATED FROM 18 TO 61 YEARS AND FOUR WERE FEMALES. THE MEAN AGE AT ONSET OF SYMPTOMS WAS 29 YEARS AND FOUR HAD A FAMILY HISTORY OF THE DISEASE. THE USUAL PRESENTING COMPLAINT WAS ARM OR SHOULDER ASYMMETRIC WEAKNESS. FOUR PATIENTS HAD BONE PAIN. FACIAL INVOLVEMENT WAS PRESENT IN FOUR. A GENETIC STUDY WAS DONE IN FIVE PATIENTS, THE OTHER TWO PATIENTS WERE RELATIVES, CONFIRMING THE CONTRACTION OR LOWER NUMBER OF REPETITIONS IN D4Z4 REGION. AFTER 12 YEARS OF FOLLOW UP ONLY 2 PATIENTS OLDER THAN 60 YEARS CANNOT WORK AND ONE FEMALE PATIENTS IS IN A SEMI DEPENDENT STATE AT THE AGE OF 30. CONCLUSIONS: THE CLINICAL WORKUP IN THE DIAGNOSIS AND THE TIMELY INDICATION OF GENETIC STUDIES ARE HIGHLIGHTED, TO AVOID UNNECESSARY AND INVASIVE PROCEDURES. THE VARIABILITY IN THE PHENOTYPIC EXPRESSION IN A SIMILAR GENETIC DEFECT IS DISCUSSED AND THE GENETIC OR EPIGENETIC MECHANISMS OF THIS MUSCULAR DYSTROPHY ARE DESCRIBED. 2015 10 3424 29 HUMAN STEM CELLS ISOLATED FROM ADULT SKELETAL MUSCLE DIFFERENTIATE INTO NEURAL PHENOTYPES. MULTIPOTENT NEURAL STEM CELLS HAVE BEEN ISOLATED FROM THE ADULT [KIRSCHENBAUM B, NEDERGAARD M, PREUSS A, BARAMI K, FRASER RA, GOLDMAN SA. IN VITRO NEURONAL PRODUCTION AND DIFFERENTIATION BY PRECURSOR CELLS DERIVED FROM THE ADULT HUMAN FOREBRAIN. CEREB CORTEX 1994;4(6):576-89; LAYWELL ED, KUKEKOV VG, STEINDLER DA. MULTIPOTENT NEUROSPHERES CAN BE DERIVED FROM FOREBRAIN SUBEPENDYMAL ZONE AND SPINAL CORD OF ADULT MICE AFTER PROTRACTED POSTMORTEM INTERVALS. EXP NEUROL 1999;156:430-3; PLUCHINO S, QUATTRINI A, BRAMBILLA E, GRITTI A, SALANI G, DINA G, ET AL. INJECTION OF ADULT NEUROSPHERES INDUCES RECOVERY IN A CHRONIC MODEL OF MULTIPLE SCLEROSIS. NATURE 2003;422:688-94] AND EMBRYONIC [VESCOVI AL, PARATI EA, GRITTI A, POULIN P, FERRARIO M, WANKE E, ET AL. ISOLATION AND CLONING OF MULTIPOTENTIAL STEM CELLS FROM THE EMBRYONIC HUMAN CNS AND ESTABLISHMENT OF TRANSPLANTABLE HUMAN NEURAL STEM CELL LINES BY EPIGENETIC STIMULATION. EXP NEUROL 1999;156:71-83] CENTRAL NERVOUS SYSTEM (CNS). IN ADDITION, NEURAL CELLS CAN BE OBTAINED FROM SOURCES OTHER THAN THE CNS BY DIFFERENTIATING STEM CELLS FROM A NON-NEURAL SOURCE DOWN A NEURAL LINEAGE. THIS HAS PREVIOUSLY BEEN PERFORMED WITH PLURIPOTENT EMBRYONIC STEM CELLS AND ADULT STEM CELLS DERIVED FROM RAT BONE MARROW [WOODBURY D, SCHWARZ EJ, PROCKOP DJ, BLACK IB. ADULT RAT AND HUMAN BONE MARROW STROMAL CELLS DIFFERENTIATE INTO NEURONS. J NEUROSCI RES 2000;61:364-70; WOODBURY D, REYNOLDS K, BLACK IB. ADULT BONE MARROW STROMAL STEM CELLS EXPRESS GERMLINE, ECTODERMAL, ENDODERMAL, AND MESODERMAL GENES PRIOR TO NEUROGENESIS. J NEUROSCI 2002;69(6):908-17] AND SKELETAL MUSCLE [ROMERO-RAMOS M, VOURC'H P, YOUNG HE, LUCAS PA, WU Y, CHIVATAKARN O, ET AL. NEURONAL DIFFERENTIATION OF STEM CELLS ISOLATED FROM ADULT MUSCLE. J NEUROSCI RES 2002;69:894-907]. PREVIOUSLY, WE HAVE ISOLATED ADULT STEM CELLS FROM HUMAN SKELETAL MUSCLE WITH THE POTENTIAL TO DIFFERENTIATE INTO MESODERM, ECTODERM, AND ENDODERM. THE FOLLOWING IN VITRO EXPERIMENTS WERE DESIGNED TO DETERMINE WHETHER HUMAN ADULT STEM CELLS BEHAVED SIMILARLY TO RAT ADULT STEM CELLS WHEN BOTH WERE ISOLATED FROM SKELETAL MUSCLE BY THE SAME PROCEDURE [ROMERO-RAMOS M, VOURC'H P, YOUNG HE, LUCAS PA, WU Y, CHIVATAKARN O, ET AL. NEURONAL DIFFERENTIATION OF STEM CELLS ISOLATED FROM ADULT MUSCLE. J NEUROSCI RES 2002;69:894-907] AND SUBJECTED TO THE SAME PROTOCOLS TO INDUCE NEUROGENESIS. THE NEURAL PHENOTYPES THAT WERE CREATED THROUGH THE NEUROCOCKTAIL OR NEUROSPHERE PROTOCOL WERE ANALYZED FOR NEURAL CHARACTERISTICS THROUGH MORPHOLOGY AND IMMUNOHISTOCHEMISTRY ANTIBODY LABELING FOR PROTEINS TO NEURONS (RT-97, BETA-TUBULIN III, NF-160, NF-200, AND SYNAPSIN), OLIGODENDROCYTES (CNPASE AND RIP), AND ASTROCYTES (GFAP). A CALCIUM UPTAKE ASSAY ALSO SHOWED RESPONSE TO THE NEURONAL EXCITOTOXIC AGENT GLUTAMIC ACID. IN CONCLUSION, THE NEURAL DIFFERENTIATED STEM CELLS DERIVED FROM ADULT SKELETAL MUSCLE MAY BE A LESS INVASIVE ALTERNATIVE FOR THE TREATMENT OF CNS DISORDERS OVER CNS DERIVED NEURAL STEM CELLS. 2006 11 6578 23 TREATMENT WITH TRICHURIS SUIS SOLUBLE PRODUCTS DURING MONOCYTE-TO-MACROPHAGE DIFFERENTIATION REDUCES INFLAMMATORY RESPONSES THROUGH EPIGENETIC REMODELING. HELMINTHS HAVE STRONG IMMUNOREGULATORY PROPERTIES THAT MAY BE EXPLOITED IN TREATMENT OF CHRONIC IMMUNE DISORDERS, SUCH AS MULTIPLE SCLEROSIS AND INFLAMMATORY BOWEL DISEASE. ESSENTIAL PLAYERS IN THE PATHOGENESIS OF THESE DISEASES ARE PROINFLAMMATORY MACROPHAGES. WE PRESENT EVIDENCE THAT HELMINTHS MODULATE THE FUNCTION AND PHENOTYPE OF THESE INNATE IMMUNE CELLS. WE FOUND THAT SOLUBLE PRODUCTS DERIVED FROM THE TRICHURIS SUIS (TSSP) SIGNIFICANTLY AFFECT THE DIFFERENTIATION OF MONOCYTES INTO MACROPHAGES AND THEIR SUBSEQUENT POLARIZATION. TSSPS REDUCE THE EXPRESSION AND PRODUCTION OF INFLAMMATORY CYTOKINES, INCLUDING IL-6 AND TNF, IN HUMAN PROINFLAMMATORY M1 MACROPHAGES. TSSPS INDUCE A CONCOMITANT ANTI-INFLAMMATORY M2 SIGNATURE, WITH INCREASED IL-10 PRODUCTION. FURTHERMORE, THEY SUPPRESS CHIT ACTIVITY AND ENHANCE SECRETION OF MATRIX METALLOPROTEINASE 9. SHORT-TERM TRIGGERING OF MONOCYTES WITH TSSPS EARLY DURING MONOCYTE-TO-MACROPHAGE DIFFERENTIATION IMPRINTED THESE PHENOTYPIC ALTERATIONS, SUGGESTING LONG-LASTING EPIGENETIC CHANGES. THE TSSP-INDUCED EFFECTS IN M1 MACROPHAGES WERE COMPLETELY REVERSED BY INHIBITING HISTONE DEACETYLASES, WHICH CORRESPONDED WITH DECREASED HISTONE ACETYLATION AT THE TNF AND IL6 PROMOTERS. THESE RESULTS DEMONSTRATE THAT TSSPS HAVE A POTENT AND SUSTAINED IMMUNOMODULATORY EFFECT ON HUMAN MACROPHAGE DIFFERENTIATION AND POLARIZATION THROUGH EPIGENETIC REMODELING AND PROVIDE NEW INSIGHTS INTO THE MECHANISMS BY WHICH HELMINTHS MODULATE HUMAN IMMUNE RESPONSES.-HOEKSEMA, M. A., LAAN, L. C., POSTMA, J. J., CUMMINGS, R. D., DE WINTHER, M. P. J., DIJKSTRA, C. D., VAN DIE, I., KOOIJ, G. TREATMENT WITH TRICHURIS SUIS SOLUBLE PRODUCTS DURING MONOCYTE-TO-MACROPHAGE DIFFERENTIATION REDUCES INFLAMMATORY RESPONSES THROUGH EPIGENETIC REMODELING. 2016 12 6657 27 UPREGULATED KAT7 IN SYNOVIAL FIBROBLASTS PROMOTES TH17 CELL DIFFERENTIATION AND INFILTRATION IN RHEUMATOID ARTHRITIS. RHEUMATOID ARTHRITIS (RA) IS A CHRONIC AUTOIMMUNE DISEASE INVOLVING MULTIPLE CELLULAR PARTICIPANTS, OF WHICH SYNOVIAL FIBROBLASTS (SFS) ARE TIGHTLY CONNECTED WITH THE DEVELOPMENT AND PROGRESSION OF RA. HERE, WE PROVIDE EVIDENCE CONFIRMING THAT KAT7, AN H4-SPECIFIC HISTONE ACETYLASE, IS UPREGULATED IN SFS OF RA PATIENTS, WHICH IS AT LEAST ATTRIBUTED TO THE STIMULATION BY RA-ASSOCIATED PROINFLAMMATORY CYTOKINES, SUCH AS TNF-ALPHA, IL-1BETA OR IFN-GAMMA. IN ADDITION, KAT7 OVEREXPRESSION IN CULTURED HUMAN FIBROBLAST-LIKE SYNOVIOCYTES (HFLSS) INDUCES IL-6 AND TGF-BETA EXPRESSION THROUGH AN EPIGENETIC MECHANISM, AND IN VITRO T HELPER 17 (TH17) CELL POLARIZATION CULTURED IN THESE SUPERNATANTS SHOWS PROMOTED CELL DIFFERENTIATION. MOREOVER, KAT7 OVEREXPRESSION IN HFLSS INDUCES CCL20 EXPRESSION VIA P44/42 MAPK PATHWAY, WHEREBY PROMOTING TH17 CELL MIGRATION. THESE TWO ACTIVITIES OF KAT7 IN RA SFS INDICATE ITS POTENTIAL ROLES IN ACCELERATING RA PATHOLOGY. OVERALL, THESE RESULTS DEMONSTRATE SOME CONNECTIONS BETWEEN KAT7 UPREGULATED IN RA SFS AND RA PROGRESSION AND PRESENT THE INHIBITION OF KAT7 ACTIVITY AS A NOVEL THERAPEUTIC TARGET FOR INTERFERING RA DISEASE. 2017 13 4146 22 MECHANISMS REGULATING MUSCLE PROTEIN SYNTHESIS IN CKD. BACKGROUND: CKD INDUCES LOSS OF MUSCLE PROTEINS PARTLY BY SUPPRESSING MUSCLE PROTEIN SYNTHESIS. MUSCLES OF MICE WITH CKD HAVE INCREASED EXPRESSION OF NUCLEOLAR PROTEIN 66 (NO66), AS DO MUSCLE BIOPSY SPECIMENS FROM PATIENTS WITH CKD OR THOSE UNDERGOING HEMODIALYSIS. INFLAMMATION STIMULATES NO66 EXPRESSION AND CHANGES IN NF-KAPPAB MEDIATE THE RESPONSE. METHODS: SUBTOTAL NEPHRECTOMY CREATED A MOUSE MODEL OF CKD WITH BUN >80 MG/DL. CROSSING NO66(FLOX/FLOX) WITH MCK-CRE MICE BRED MUSCLE-SPECIFIC NO66 (MCK-NO66) KNOCKOUT MICE. EXPERIMENTS ASSESSED THE EFFECT OF REMOVING NO66. RESULTS: MUSCLE-SPECIFIC NO66 KNOCKOUT IN MICE BLOCKS CKD-INDUCED LOSS OF MUSCLE MASS AND IMPROVES PROTEIN SYNTHESIS. NO66 SUPPRESSION OF RIBOSOMAL BIOGENESIS VIA DEMETHYLASE ACTIVITY IS THE MECHANISM BEHIND THESE RESPONSES. IN MUSCLE CELLS, EXPRESSION OF NO66, BUT NOT OF DEMETHYLASE-DEAD MUTANT NO66, DECREASED H3K4ME3 AND H3K36ME3 AND SUPPRESSED PRE-RRNA EXPRESSION. KNOCKING OUT NO66 INCREASED THE ENRICHMENT OF H3K4ME3 AND H3K36ME3 ON RIBOSOMAL DNA. IN PRIMARY MUSCLE CELLS AND IN MUSCLES OF MICE WITHOUT NO66, RIBOSOMAL RNA, PRE-RRNA, AND PROTEIN SYNTHESIS ALL INCREASED. CONCLUSIONS: CKD SUPPRESSES MUSCLE PROTEIN SYNTHESIS VIA EPIGENETIC MECHANISMS THAT NO66 MEDIATES. BLOCKING NO66 COULD SUGGEST STRATEGIES THAT COUNTER CKD-INDUCED ABNORMAL MUSCLE PROTEIN CATABOLISM. 2020 14 4042 33 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 15 5017 22 PERSISTENT INFECTION OF CULTURED CELLS WITH MOUSE HEPATITIS VIRUS (MHV) RESULTS FROM THE EPIGENETIC EXPRESSION OF THE MHV RECEPTOR. THE A59 STRAIN OF MURINE CORONAVIRUS MOUSE HEPATITIS VIRUS (MHV) CAN CAUSE PERSISTENT INFECTION OF 17C1-1 CELLS AND OTHER MURINE CELL LINES. PERSISTENTLY INFECTED CULTURES RELEASED LARGE AMOUNTS OF VIRUS (10(7) TO 10(8) PFU/ML) AND WERE RESISTANT TO SUPERINFECTION WITH MHV BUT NOT TO INFECTION WITH UNRELATED SEMLIKI FOREST AND VESICULAR STOMATITIS VIRUSES. THE CULTURE MEDIUM FROM PERSISTENTLY INFECTED CULTURES DID NOT CONTAIN A SOLUBLE INHIBITOR SUCH AS INTERFERON THAT PROTECTED UNINFECTED CELLS FROM INFECTION BY MHV OR VESICULAR STOMATITIS VIRUS. THE PERSISTENT INFECTION WAS CURED IF FEWER THAN 100 CELLS WERE TRANSFERRED DURING SUBCULTURING, AND SUCH CURED CULTURES WERE SUSCEPTIBLE TO REINFECTION AND THE REESTABLISHMENT OF PERSISTENT INFECTION. CULTURES OF 17C1-1 CELLS THAT HAD BEEN NEWLY CLONED FROM SINGLE CELLS CONSISTED OF A MIXTURE OF MHV-RESISTANT AND -SUSCEPTIBLE CELLS. 17C1-1/#97 CELLS, WHICH WERE CURED BY SUBCLONING AFTER 97 PASSAGES OF A PERSISTENTLY INFECTED CULTURE OVER A 1-YEAR PERIOD, CONTAINED 5 TO 10% OF THEIR POPULATION AS SUSCEPTIBLE CELLS, WHILE 17C1-1/#402 CELLS, WHICH WERE CURED BY SUBCLONING AFTER 402 PASSAGES OVER A 3-YEAR PERIOD, HAD LESS THAN 1% SUSCEPTIBLE CELLS. SUSCEPTIBILITY TO INFECTION CORRELATED WITH THE EXPRESSION OF MHV RECEPTOR GLYCOPROTEIN (MHVR [BGP1A]). FLUORESCENCE-ACTIVATED CELL SORTER ANALYSIS WITH ANTIBODY TO MHVR SHOWED THAT 17C1-1/#97 CELLS CONTAINED A SMALL FRACTION OF MHVR-EXPRESSING CELLS. THESE MHVR-EXPRESSING CELLS WERE SELECTIVELY ELIMINATED WITHIN 24 H AFTER CHALLENGE WITH MHV-A59, AND PRETREATMENT OF 17C1-1/#97 CELLS WITH MONOCLONAL ANTIBODY CC1, WHICH BINDS TO THE N-TERMINAL DOMAIN OF MHVR, BLOCKED INFECTION. WE CONCLUDE THAT THE SUBPOPULATION OF MHVR-EXPRESSING CELLS WERE INFECTED AND KILLED IN ACUTELY OR PERSISTENTLY INFECTED CULTURES, WHILE THE SUBPOPULATION OF MHVR-NONEXPRESSING CELLS SURVIVED AND PROLIFERATED. THE SUBPOPULATION OF MHVR-NEGATIVE CELLS PRODUCED A SMALL PROPORTION OF PROGENY CELLS THAT EXPRESSED MHVR AND BECAME INFECTED, THEREBY MAINTAINING THE PERSISTENT INFECTION AS A STEADY-STATE CARRIER CULTURE. THUS, IN 17C1-1 CELL CULTURES, THE UNSTABLE OR EPIGENETIC EXPRESSION OF MHVR PERMITTED THE ESTABLISHMENT OF A PERSISTENT, CHRONIC INFECTION. 1995 16 1335 26 DERMAL FIBROBLASTS CULTURED FROM DONORS WITH TYPE 2 DIABETES MELLITUS RETAIN AN EPIGENETIC MEMORY ASSOCIATED WITH POOR WOUND HEALING RESPONSES. THE PREVALENCE OF TYPE 2 DIABETES MELLITUS (T2DM) IS ESCALATING GLOBALLY. PATIENTS SUFFER FROM MULTIPLE COMPLICATIONS INCLUDING THE DEVELOPMENT OF CHRONIC WOUNDS THAT CAN LEAD TO AMPUTATION. THESE WOUNDS ARE CHARACTERISED BY AN INFLAMMATORY ENVIRONMENT INCLUDING ELEVATED TUMOUR NECROSIS FACTOR ALPHA (TNF-ALPHA). DERMAL FIBROBLASTS (DF) ARE CRITICAL FOR EFFECTIVE WOUND HEALING, SO WE SOUGHT TO ESTABLISH WHETHER THERE WERE ANY DIFFERENCES IN DF CULTURED FROM T2DM DONORS OR THOSE WITHOUT DIABETES (ND-DF). ND- AND T2DM-DF WHEN CULTURED SIMILARLY IN VITRO SECRETED COMPARABLE CONCENTRATIONS OF TNF-ALPHA. FUNCTIONALLY, PRE-TREATMENT WITH TNF-ALPHA REDUCED THE PROLIFERATION OF ND-DF AND TRANSIENTLY ALTERED ND-DF MORPHOLOGY; HOWEVER, T2DM-DF WERE RESISTANT TO THESE TNF-ALPHA INDUCED CHANGES. IN CONTRAST, TNF-ALPHA INHIBITED ND- AND T2DM-DF MIGRATION AND MATRIX METALLOPROTEASE EXPRESSION TO THE SAME DEGREE, ALTHOUGH T2DM-DF EXPRESSED SIGNIFICANTLY HIGHER LEVELS OF TISSUE INHIBITOR OF METALLOPROTEASES (TIMP)-2. FINALLY, TNF-ALPHA SIGNIFICANTLY INCREASED THE SECRETION OF PRO-INFLAMMATORY CYTOKINES (INCLUDING CCL2, CXCL1 AND SERPINE1) IN ND-DF, WHILST THIS EFFECT IN T2DM-DF WAS BLUNTED, PRESUMABLY DUE TO THE TENDENCY TO HIGHER BASELINE PRO-INFLAMMATORY CYTOKINE EXPRESSION OBSERVED IN THIS CELL TYPE. COLLECTIVELY, THESE DATA DEMONSTRATE THAT T2DM-DF EXHIBIT A SELECTIVE LOSS OF RESPONSIVENESS TO TNF-ALPHA, PARTICULARLY REGARDING PROLIFERATIVE AND SECRETORY FUNCTIONS. THIS HIGHLIGHTS IMPORTANT PHENOTYPIC CHANGES IN T2DM-DF THAT MAY EXPLAIN THE SUSCEPTIBILITY TO CHRONIC WOUNDS IN THESE PATIENTS. 2021 17 916 26 CHRONIC HIGH GLUCOSE AND INSULIN STIMULATE BONE-MARROW STROMAL CELLS ADIPOGENIC DIFFERENTIATION IN YOUNG SPONTANEOUSLY HYPERTENSIVE RATS. WE EVALUATED WHETHER GENETIC PREDISPOSITION IS SUFFICIENT TO INDUCE CHANGES DUE TO CHRONIC HIGH GLUCOSE (HG; 25 MMOL/L) IN THE PRESENCE OR ABSENCE OF INSULIN (HGI; 10 MUG/ML) ON OSTEOGENIC DIFFERENTIATION AND MARKERS IN BONE-MARROW MESENCHYMAL STEM CELLS (BMSCS) FROM YOUNG WISTAR (WBMSCS) AND SPONTANEOUS HYPERTENSIVE RATS (SBMSCS) WITHOUT HYPERTENSION. HG SUPPRESSED OSTEOGENIC DIFFERENTIATION IN BOTH THE STRAINS, OBSERVED BY MINERALIZATION INHIBITION AND DECREASED LEVELS OF THE OSTEOGENIC MARKERS RUNX2, OSTERIX, OSTEOPONTIN, AND BONE SIALOPROTEIN, COMPARED TO OSTEOGENIC MEDIUM (OM) CELLS. IN WBMSCS, THE EFFECTS OF HG WERE ASSOCIATED WITH THE DOWN REGULATION OF ERK1/2 AND UP REGULATION OF P38 ACTIVITIES; HOWEVER, HGI DID NOT REVERT THE EFFECTS OF HG ON MAPK ACTIVITIES. MOREOVER, HG DID NOT AFFECT MAPK SIGNALING IN SBMSCS COMPARED TO THAT IN OM. HGI INCREASED MINERALIZATION IN WBMSCS COMPARED TO THAT IN OM, BUT NOT IN SBMSCS. HIGH EXPRESSION OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR-GAMMA AND GLUCOSE TRANSPORTER TYPE 4 IN OM COULD BE RELATED WITH THE PREDISPOSITION TO ADIPOGENIC DIFFERENTIATION NOTED IN SBMSCS AND WAS CONFIRMED BY EMERGENCE OF ADIPOCYTE-LIKE CELLS BY HGI TREATMENT. DOWNREGULATION OF P38 AND UPREGULATION OF JNK ACTIVITIES WERE OBSERVED IN BOTH BMSCS TREATED WITH HGI COMPARED TO THOSE TREATED BY HG. MA (OSMOTIC CONTROL) ALSO SUPPRESSED OSTEOGENIC DIFFERENTIATION IN BOTH THE STRAINS. IN CONCLUSION, WE DEMONSTRATED THAT SBMSCS FROM YOUNG SPONTANEOUS HYPERTENSIVE RATS, WITHOUT HYPERTENSION BUT WITH GENETIC AND EPIGENETIC PREDISPOSITION, EXHIBITED DECREASED OSTEOBLASTIC DIFFERENTIATION UNDER HG AND HGI DID NOT REVERT THE EFFECTS OF HG IN SBMSCS BUT INCREASED ADIPOGENIC DIFFERENTIATION. 2018 18 3716 21 INHIBITING INFLAMMATION WITH MYELOID CELL-SPECIFIC NANOBIOLOGICS PROMOTES ORGAN TRANSPLANT ACCEPTANCE. INDUCING GRAFT ACCEPTANCE WITHOUT CHRONIC IMMUNOSUPPRESSION REMAINS AN ELUSIVE GOAL IN ORGAN TRANSPLANTATION. USING AN EXPERIMENTAL TRANSPLANTATION MOUSE MODEL, WE DEMONSTRATE THAT LOCAL MACROPHAGE ACTIVATION THROUGH DECTIN-1 AND TOLL-LIKE RECEPTOR 4 (TLR4) DRIVES TRAINED IMMUNITY-ASSOCIATED CYTOKINE PRODUCTION DURING ALLOGRAFT REJECTION. WE CONDUCTED NANOIMMUNOTHERAPEUTIC STUDIES AND FOUND THAT A SHORT-TERM MTOR-SPECIFIC HIGH-DENSITY LIPOPROTEIN (HDL) NANOBIOLOGIC TREATMENT (MTORI-HDL) AVERTED MACROPHAGE AEROBIC GLYCOLYSIS AND THE EPIGENETIC MODIFICATIONS UNDERLYING INFLAMMATORY CYTOKINE PRODUCTION. THE RESULTING REGULATORY MACROPHAGES PREVENTED ALLOREACTIVE CD8(+) T CELL-MEDIATED IMMUNITY AND PROMOTED TOLEROGENIC CD4(+) REGULATORY T (TREG) CELL EXPANSION. TO ENHANCE THERAPEUTIC EFFICACY, WE COMPLEMENTED THE MTORI-HDL TREATMENT WITH A CD40-TRAF6-SPECIFIC NANOBIOLOGIC (TRAF6I-HDL) THAT INHIBITS CO-STIMULATION. THIS SYNERGISTIC NANOIMMUNOTHERAPY RESULTED IN INDEFINITE ALLOGRAFT SURVIVAL. TOGETHER, WE SHOW THAT HDL-BASED NANOIMMUNOTHERAPY CAN BE EMPLOYED TO CONTROL MACROPHAGE FUNCTION IN VIVO. OUR STRATEGY, FOCUSED ON PREVENTING INFLAMMATORY INNATE IMMUNE RESPONSES, PROVIDES A FRAMEWORK FOR DEVELOPING TARGETED THERAPIES THAT PROMOTE IMMUNOLOGICAL TOLERANCE. 2018 19 1777 32 EDIBLE BLUE-GREEN ALGAE REDUCE THE PRODUCTION OF PRO-INFLAMMATORY CYTOKINES BY INHIBITING NF-KAPPAB PATHWAY IN MACROPHAGES AND SPLENOCYTES. BACKGROUND: CHRONIC INFLAMMATION CONTRIBUTES TO THE DEVELOPMENT OF PATHOLOGICAL DISORDERS INCLUDING INSULIN RESISTANCE AND ATHEROSCLEROSIS. IDENTIFICATION OF ANTI-INFLAMMATORY NATURAL PRODUCTS CAN PREVENT THE INFLAMMATORY DISEASES. METHODS: ANTI-INFLAMMATORY EFFECTS OF BLUE-GREEN ALGAE (BGA), I.E., NOSTOC COMMUNE VAR. SPHAEROIDES KUTZING (NO) AND SPIRULINA PLATENSIS (SP), WERE COMPARED IN RAW 264.7 AND MOUSE BONE MARROW-DERIVED MACROPHAGES (BMM) AS WELL AS SPLENOCYTES FROM APOLIPOPROTEIN E KNOCKOUT (APOE(-/-)) MICE FED BGA. RESULTS: WHEN MACROPHAGES PRETREATED WITH 100MUG/ML NO LIPID EXTRACT (NOE) OR SP LIPID EXTRACT (SPE) WERE ACTIVATED BY LIPOPOLYSACCHARIDE (LPS), EXPRESSION AND SECRETION OF PRO-INFLAMMATORY CYTOKINES, SUCH AS TUMOR NECROSIS FACTOR ALPHA (TNFALPHA), INTERLEUKIN 1BETA (IL-1BETA), AND IL-6, WERE SIGNIFICANTLY REPRESSED. NOE AND SPE ALSO SIGNIFICANTLY REPRESSED THE EXPRESSION OF TNFALPHA AND IL-1BETA IN BMM. LPS-INDUCED SECRETION OF IL-6 WAS LOWER IN SPLENOCYTES FROM APOE(-/-) FED AN ATHEROGENIC DIET CONTAINING 5% NO OR SP FOR 12WEEKS. IN RAW 264.7 MACROPHAGES, NOE AND SPE MARKEDLY DECREASED NUCLEAR TRANSLOCATION OF NF-KAPPAB. THE DEGREE OF REPRESSION OF PRO-INFLAMMATORY GENE EXPRESSION BY ALGAL EXTRACTS WAS MUCH STRONGER THAN THAT OF SN50, AN INHIBITOR OF NF-KAPPAB NUCLEAR TRANSLOCATION. TRICHOSTATIN A, A PAN HISTONE DEACETYLASE INHIBITOR, INCREASED BASAL EXPRESSION OF IL-1BETA AND ATTENUATED THE REPRESSION OF THE GENE EXPRESSION BY SPE. SPE SIGNIFICANTLY DOWN-REGULATED MRNA ABUNDANCE OF 11 HDAC ISOFORMS, CONSEQUENTLY INCREASING ACETYLATED HISTONE 3 LEVELS. CONCLUSION: NOE AND SPE REPRESS PRO-INFLAMMATORY CYTOKINE EXPRESSION AND SECRETION IN MACROPHAGES AND SPLENOCYTES VIA INHIBITION OF NF-KAPPAB PATHWAY. HISTONE ACETYLATION STATE IS LIKELY INVOLVED IN THE INHIBITION. GENERAL SIGNIFICANCE: THIS STUDY UNDERSCORES NATURAL PRODUCTS CAN EXERT ANTI-INFLAMMATORY EFFECTS BY EPIGENETIC MODIFICATIONS SUCH AS HISTONE ACETYLATION. 2013 20 6586 28 TUBASTATIN, A SELECTIVE HISTONE DEACETYLASE 6 INHIBITOR SHOWS ANTI-INFLAMMATORY AND ANTI-RHEUMATIC EFFECTS. EPIGENETIC MODIFICATIONS REPRESENT A PROMISING NEW APPROACH TO MODULATE CELL FUNCTIONS AS OBSERVED IN AUTOIMMUNE DISEASES. EMERGING EVIDENCE SUGGESTS THE UTILITY OF HDAC INHIBITORS IN THE TREATMENT OF CHRONIC IMMUNE AND INFLAMMATORY DISORDERS. HOWEVER, CLASS AND ISOFORM SELECTIVE INHIBITION OF HDAC IS CURRENTLY FAVORED AS IT LIMITS THE TOXICITY THAT HAS BEEN OBSERVED WITH PAN-HDAC INHIBITORS. HDAC6, A MEMBER OF THE HDAC FAMILY, WHOSE MAJOR SUBSTRATE IS ALPHA-TUBULIN, IS BEING INCREASINGLY IMPLICATED IN THE PATHOGENESIS OF INFLAMMATORY DISORDERS. THE PRESENT STUDY WAS CARRIED OUT TO STUDY THE POTENTIAL ANTI-INFLAMMATORY AND ANTI-RHEUMATIC EFFECTS OF HDAC6 SELECTIVE INHIBITOR TUBASTATIN. TUBASTATIN, A POTENT HUMAN HDAC6 INHIBITOR WITH AN IC50 OF 11 NM SHOWED SIGNIFICANT INHIBITION OF TNF-ALPHA AND IL-6 IN LPS STIMULATED HUMAN THP-1 MACROPHAGES WITH AN IC50 OF 272 NM AND 712 NM RESPECTIVELY. ADDITIONALLY, TUBASTATIN INHIBITED NITRIC OXIDE (NO) SECRETION IN MURINE RAW 264.7 MACROPHAGES DOSE DEPENDENTLY WITH AN IC50 OF 4.2 MUM AND INDUCED ALPHA-TUBULIN HYPERACETYLATION CORRESPONDING TO HDAC6 INHIBITION IN THP-1 CELLS WITHOUT AFFECTING THE CELL VIABILITY. TUBASTATIN SHOWED SIGNIFICANT INHIBITION OF PAW VOLUME AT 30 MG/KG I.P. IN A FREUND'S COMPLETE ADJUVANT (FCA) INDUCED ANIMAL MODEL OF INFLAMMATION. THE DISEASE MODIFYING ACTIVITY OF TUBASTATIN WAS ALSO EVIDENT IN COLLAGEN INDUCED ARTHRITIS DBA1 MOUSE MODEL AT 30 MG/KG I.P. THE SIGNIFICANT ATTENUATION OF CLINICAL SCORES (~70%) BY TUBASTATIN WAS CONFIRMED HISTOPATHOLOGICALLY AND WAS FOUND COMPARABLE TO DEXAMETHASONE (~90% INHIBITION OF CLINICAL SCORES). TUBASTATIN SHOWED SIGNIFICANT INHIBITION OF IL-6 IN PAW TISSUES OF ARTHRITIC MICE. THE PRESENT WORK HAS DEMONSTRATED ANTI-INFLAMMATORY AND ANTIRHEUMATIC EFFECTS OF A SELECTIVE HDAC6 INHIBITOR TUBASTATIN. 2013