1 814 143 CHANGES IN SMALL NONCODING RNA EXPRESSION DURING CHONDROCYTE SENESCENCE. OBJECTIVE: OSTEOARTHRITIS (OA) IS CHARACTERIZED BY THE CHRONIC AND PROGRESSIVE DETERIORATION OF ARTICULAR CARTILAGE. CHONDROCYTE SENESCENCE COULD LEAD TO A SHIFT IN THE BALANCE BETWEEN EXTRACELLULAR MATRIX (ECM) COMPONENT SYNTHESIS AND DEGRADATION. SMALL NONCODING RNAS (SNCRNAS), INCLUDING MICRORNAS (MIRNAS), P-ELEMENT-INDUCED WIMPY TESTIS-(PIWI-) INTERACTING RNAS (PIRNAS), SMALL NUCLEOLAR RNAS (SNORNAS), SMALL NUCLEAR RNAS (SNRNAS), AND REPEAT-ASSOCIATED SIRNAS (RASIRNAS), ARE A CLASS OF IMPORTANT EPIGENETIC MOLECULES. WE AIMED TO GAIN INSIGHTS INTO THE CHANGES AND ROLES OF SNCRNA IN CHONDROCYTE SENESCENCE. DESIGN: HEALTHY MOUSE POSTNATAL CHONDROCYTES WERE ISOLATED, AND A REPLICATIVE AGING MODEL WAS CONSTRUCTED. WE USED SMALL RNA SEQUENCING (SMALL RNA-SEQ) TO GENERATE EXTENSIVE SMALL RNA DATA. WE IDENTIFIED DIFFERENTIALLY EXPRESSED SNCRNAS AND PERFORMED TISSUE-SPECIFIC ANALYSIS USING REAL-TIME QUANTITATIVE POLYMERASE CHAIN REACTION (QRT-PCR). BETA-GALACTOSIDASE STAINING WAS USED TO DETECT CHONDROCYTE SENESCENCE. THE RESULTS SHOWED THAT THE EXPRESSION PROFILES OF SNCRNA IN PASSAGE 5 CHONDROCYTES WERE SIGNIFICANTLY DIFFERENT FROM THOSE IN PASSAGE 0 CHONDROCYTES. THE EXPRESSION OF SNCRNA WAS TISSUE SPECIFIC. WE FOUND THAT 40 MIRNAS WERE UPREGULATED AND 70 MIRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT MIR-132-5P EXPRESSION INHIBITION PREVENTED CHONDROCYTE SENESCENCE. WE FOUND THAT 8 PIRNAS WERE UPREGULATED AND 17 PIRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT PIRNA PIR_025576 OVEREXPRESSION DELAYED CHONDROCYTE SENESCENCE. WE FOUND THAT 24 SNORNAS WERE UPREGULATED AND 28 SNORNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT SNORNA ENSMUSG00000087935 OVEREXPRESSION DELAYED CHONDROCYTE SENESCENCE. WE FOUND THAT 5 SNRNAS WERE UPREGULATED AND 6 SNRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE, AND THAT SNRNA ENSMUSG00000064682 OVEREXPRESSION DELAYED CHONDROCYTE SENESCENCE. WE FOUND THAT 1 RASIRNA WAS UPREGULATED AND 4 RASIRNAS WERE DOWNREGULATED DURING CHONDROCYTE SENESCENCE. CONCLUSIONS: THESE FINDINGS MIGHT PROVIDE NOVEL INSIGHTS INTO OA PATHOGENESIS AND CONTRIBUTE TO THE DEVELOPMENT OF CANDIDATES FOR TARGETED THERAPEUTICS IN OA. 2022 2 1063 21 CLINICAL SIGNIFICANCE OF MICRORNAS IN CHRONIC AND ACUTE HUMAN LEUKEMIA. SMALL NON-CODING MICRORNAS (MIRNAS) ARE EPIGENETIC REGULATORS THAT TARGET SPECIFIC CELLULAR MRNA TO MODULATE GENE EXPRESSION PATTERNS AND CELLULAR SIGNALING PATHWAYS. MIRNAS ARE INVOLVED IN A WIDE RANGE OF BIOLOGICAL PROCESSES AND ARE FREQUENTLY DEREGULATED IN HUMAN CANCERS. NUMEROUS MIRNAS PROMOTE TUMORIGENESIS AND CANCER PROGRESSION BY ENHANCING TUMOR GROWTH, ANGIOGENESIS, INVASION AND IMMUNE EVASION, WHILE OTHERS HAVE TUMOR SUPPRESSIVE EFFECTS (HAYES, ET AL., TRENDS MOL MED 20(8): 460-9, 2014; STAHLHUT AND SLACK, GENOME MED 5 (12): 111, 2013). THE EXPRESSION PROFILE OF CANCER MIRNAS CAN BE USED TO PREDICT PATIENT PROGNOSIS AND CLINICAL RESPONSE TO TREATMENT (BOUCHIE, NAT BIOTECHNOL 31(7): 577, 2013). THE MAJORITY OF MIRNAS ARE INTRACELLULAR LOCALIZED, HOWEVER CIRCULATING MIRNAS HAVE BEEN DETECTED IN VARIOUS BODY FLUIDS AND REPRESENT NEW BIOMARKERS OF SOLID AND HEMATOLOGIC CANCERS (FABRIS AND CALIN, MOL ONCOL 10(3):503-8, 2016; ALLEGRA, ET AL., INT J ONCOL 41(6): 1897-912, 2012). THIS REVIEW DESCRIBES THE CLINICAL RELEVANCE OF MIRNAS, LNCRNAS AND SNORNAS IN THE DIAGNOSIS, PROGNOSIS AND TREATMENT RESPONSE IN PATIENTS WITH CHRONIC LYMPHOCYTIC LEUKEMIA (CLL), CHRONIC MYELOID LEUKEMIA (CML), ACUTE LYMPHOCYTIC LEUKEMIA (ALL), ACUTE MYELOID LEUKEMIA (AML) AND ACUTE ADULT T-CELL LEUKEMIA (ATL). 2016 3 4179 33 MEN WHO INJECT OPIOIDS EXHIBIT ALTERED TRNA-GLY-GCC ISOFORMS IN SEMEN. IN ADDITION TO THEIR ROLE IN PROTEIN TRANSLATION, TRNAS CAN BE CLEAVED INTO SHORTER, BIOLOGICALLY ACTIVE FRAGMENTS CALLED TRNA FRAGMENTS (TRFS). SPECIFIC TRFS FROM SPERMATOCYTES CAN PROPAGATE METABOLIC DISORDERS IN SECOND GENERATIONS OF MICE. THUS, TRFS IN GERMLINE CELLS ARE A MECHANISM OF EPIGENETIC INHERITANCE. IT HAS ALSO BEEN SHOWN THAT STRESS AND TOXINS CAN CAUSE ALTERATIONS IN TRF PATTERNS. WE WERE THEREFORE INTERESTED IN WHETHER INJECTING ILLICIT DRUGS, A MAJOR STRESSOR, IMPACTS TRFS IN GERMLINE CELLS. WE SEQUENCED RNA FROM SPERMATOCYTES AND FROM SEMEN-DERIVED EXOSOMES FROM PEOPLE WHO INJECT ILLICIT DRUGS (PWID) AND FROM NON-DRUG USING CONTROLS, BOTH GROUPS OF UNKNOWN FERTILITY STATUS. ALL PWID INJECTED OPIOIDS DAILY, BUT MOST ALSO USED OTHER ILLICIT DRUGS. THE TRF CLEAVAGE PRODUCTS FROM GLY-GCC TRNA WERE MARKEDLY DIFFERENT BETWEEN SPERMATOCYTES FROM PWID COMPARED TO CONTROLS. OVER 90% OF READS IN CONTROLS MAPPED TO SHORTER GLY-GCC TRFS, WHILE IN PWID ONLY 45% DID. IN CONTRAST, ONLY 4.1% OF READS IN CONTROLS MAPPED TO A LONGER TRFS VERSUS 45.6% IN PWID. THE LONG/SHORT TRF RATIO WAS SIGNIFICANTLY HIGHER IN PWID THAN CONTROLS (0.23 VERSUS 0.16, P = 0.0128). WE ALSO REPORT DIFFERENTIAL EXPRESSION OF A GROUP OF SMALL NUCLEOLAR RNAS (SNORNAS) IN SEMEN-DERIVED EXOSOMES, INCLUDING, AMONG OTHERS, ACA14A, U19, AND U3-3. THUS, PWID EXHIBITED AN ALTERED CLEAVAGE PATTERN OF TRNA-GLY-GCC IN SPERMATOCYTES AND AN ALTERED CARGO OF SNORNAS IN SEMEN-DERIVED EXOSOMES. PARTICIPANTS WERE NOT EXCLUSIVELY USING OPIOIDS AND WERE NOT MATCHED WITH CONTROLS IN TERMS OF DIET, CHRONIC DISEASE, OR OTHER STRESSORS, SO OUR FINDING ARE NOT CONCLUSIVELY LINKED TO OPIOID USE. HOWEVER, ALL INDIVIDUALS IN THE PWID GROUP DID INJECT HEROIN DAILY. OUR STUDY INDICATES A POTENTIAL FOR OPIOID INJECTION AND/OR ITS ASSOCIATED MULTI-DRUG USE HABITS AND LIFESTYLE CHANGES TO INFLUENCE EPIGENETIC INHERITANCE. 2023 4 4364 31 MIRNA DEREGULATION BY EPIGENETIC SILENCING DISRUPTS SUPPRESSION OF THE ONCOGENE PLAG1 IN CHRONIC LYMPHOCYTIC LEUKEMIA. MICRORNAS (MIRNA) PLAY A KEY ROLE IN CELLULAR REGULATION AND, IF DEREGULATED, IN THE DEVELOPMENT OF NEOPLASTIC DISORDERS INCLUDING CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). RNAS FROM PRIMARY CELLS OF 50 TREATMENT-NAIVE CLL PATIENTS AND PERIPHERAL B CELLS OF 14 HEALTHY DONORS WERE APPLIED TO MIRNA EXPRESSION PROFILING USING BEAD CHIP TECHNOLOGY. IN CLL CELLS, A SET OF 7 UP- AND 19 DOWN-REGULATED MIRNAS WAS IDENTIFIED. AMONG THE MIRNAS DOWN-REGULATED IN CLL CELLS, 6 OF 10 MIRNA PROMOTERS EXAMINED SHOWED GAIN OF METHYLATION COMPARED WITH NORMAL B-CELL CONTROLS. SUBSEQUENT TARGET PREDICTION OF DEREGULATED MIRNAS REVEALED A HIGHLY SIGNIFICANT BINDING PREDICTION AT THE 3' UNTRANSLATED REGION OF THE PLEOMORPHIC ADENOMA GENE 1 (PLAG1) ONCOGENE. LUCIFERASE REPORTER ASSAYS INCLUDING SITE-DIRECTED MUTAGENESIS OF BINDING SITES REVEALED A SIGNIFICANT REGULATION OF PLAG1 BY MIR-181A, MIR-181B, MIR-107, AND MIR-424. ALTHOUGH EXPRESSION OF PLAG1 MRNA WAS NOT AFFECTED, PLAG1 PROTEIN EXPRESSION WAS SHOWN TO BE SIGNIFICANTLY ELEVATED IN CLL CELLS COMPARED WITH THE LEVELS IN HEALTHY DONOR B CELLS. IN SUMMARY, WE COULD DEMONSTRATE DISRUPTION OF MIRNA-MEDIATED TRANSLATIONAL CONTROL, PARTLY DUE TO EPIGENETIC TRANSCRIPTIONAL SILENCING OF MIRNAS, WITH SUBSEQUENT OVEREXPRESSION OF THE ONCOGENIC TRANSCRIPTION FACTOR PLAG1 AS A PUTATIVE NOVEL MECHANISM OF CLL PATHOGENESIS. 2009 5 25 32 A 3' TRNA-DERIVED FRAGMENT PRODUCED BY TRNA(LEUAAG) AND TRNA(LEUTAG) IS ASSOCIATED WITH POOR PROGNOSIS IN B-CELL CHRONIC LYMPHOCYTIC LEUKEMIA, INDEPENDENTLY OF CLASSICAL PROGNOSTIC FACTORS. OBJECTIVE: 3' TRNA-DERIVED FRAGMENTS (3' TRFS) ARE IMPORTANT EPIGENETIC REGULATORS IN NORMAL AND PATHOLOGICAL CONDITIONS. IN THIS STUDY, WE AIMED TO EXPLORE THE POTENTIAL VALUE OF A 3' TRF AS A PROGNOSTIC AND/OR SCREENING BIOMARKER FOR B-CELL CHRONIC LYMPHOCYTIC LEUKEMIA (B-CLL). METHODS: PUBLICLY AVAILABLE NEXT-GENERATION SEQUENCING DATA FROM 20 B-CLL CASES WERE ANALYZED, FOLLOWED BY PREDICTION OF TARGETS OF THE MOST ABUNDANTLY AND UBIQUITOUSLY EXPRESSED 3' TRFS, LEADING TO SELECTION OF TRF-LEU(AAG/TAG) . PBMCS WERE ISOLATED FROM BLOOD SAMPLES OF 91 B-CLL PATIENTS AND 43 NON-LEUKEMIC DONORS, FOLLOWED BY TOTAL RNA EXTRACTION, IN-VITRO POLYADENYLATION, AND FIRST-STRAND CDNA SYNTHESIS. NEXT, A REAL-TIME QUANTITATIVE PCR (QPCR) ASSAY WAS DEVELOPED FOR THE ACCURATE QUANTIFICATION OF TRF-LEU(AAG/TAG) AND APPLIED IN ALL SAMPLES, PRIOR TO BIOSTATISTICAL ANALYSIS. RESULTS: HIGH TRF-LEU(AAG/TAG) LEVELS ARE ASSOCIATED WITH INFERIOR OVERALL SURVIVAL (OS) OF B-CLL PATIENTS. THE UNFAVORABLE SIGNIFICANCE OF TRF-LEU(AAG/TAG) WAS INDEPENDENT OF ESTABLISHED PROGNOSTIC FACTORS IN B-CLL. STRATIFIED KAPLAN-MEIER OS ANALYSIS UNCOVERED THE UNFAVORABLE PROGNOSTIC ROLE OF HIGH TRF-LEU(AAG/TAG) LEVELS FOR PATIENTS IN BINET A OR RAI I STAGE, NEGATIVE CD38 EXPRESSION, MUTATED, OR UNMUTATED IGHV GENOMIC LOCUS. CONCLUSION: OUR APPROACH REVEALED THE INDEPENDENT PROGNOSTIC VALUE OF A PARTICULAR 3' TRF, DERIVED FROM TRNA(LEUAAG) AND TRNA(LEUTAG) (TRF-LEU(AAG/TAG) ) IN B-CLL. 2021 6 1227 29 CRITICAL ROLE OF MICRORNAS IN CHRONIC LYMPHOCYTIC LEUKEMIA: OVEREXPRESSION OF THE ONCOGENE PLAG1 BY DEREGULATED MIRNAS. MICRORNAS (MIRNAS) ARE SMALL, GENE ENCODED RNAS WHICH ARE ABLE TO INFLUENCE GENE EXPRESSION IN BINDING TO THE 3'UTR OF MRNAS. COMPARED TO HEALTHY TISSUES, THE GLOBAL EXPRESSION OF MIRNAS IN CANCEROUS TISSUE IS FREQUENTLY DOWN-REGULATED. LIKEWISE IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL), DOWN-REGULATION OF SEVERAL MIRNAS HAS BEEN REPORTED. ANALYSIS OF MIRNA PROMOTERS FOR EPIGENETIC MODIFICATIONS REVEALED A STRONGER METHYLATION OF DOWN-REGULATED MIRNAS IN CLL. TO DATE, SEVERAL TARGET GENES AFFECTED BY DEREGULATED MIRNAS HAVE BEEN IDENTIFIED THAT HAVE IMPACT ON CLL PATHOGENESIS. THE BEST-DESCRIBED CONSEQUENCE OF MIRNA DEREGULATION IS FOR MIRNA-15/16 CLUSTER DELETION, WHICH IS FREQUENTLY DOWN-REGULATED IN A SUBGROUP OF PATIENTS WITH CLL CARRYING 13Q14 DELETION. SO FAR, MODELS FOR MIRNA DEREGULATION HAVE ADDRESSED JUST SINGLE MIRNAS. FOR ASSESSMENT OF COMPLETE MIRNA DEREGULATION, FURTHER EVALUATION OF THE RESULTS FROM MICROARRAY STUDIES IS NEEDED. PREVIOUSLY WE IDENTIFIED THE ONCOGENE PLAG1, WHOSE EXPRESSION IS AFFECTED BY VARIOUS MIRNAS DEREGULATED IN CLL. THE INVOLVEMENT OF MIRNAS IN PLAG1 EXPRESSION WAS SHOWN TO BE RELEVANT IN PLEOMORPHIC ADENOMAS OF THE SALIVARY GLAND, TOO. AS PLAG1 IS HIGHLY OVEREXPRESSED, AND ITS TARGET GENES APPEAR TO BE DEREGULATED IN CLL, E.G. BCL-2, PLAG1 IS A PUTATIVE NEW RELEVANT ONCOGENE INVOLVED IN THE PATHOGENESIS OF CLL. 2010 7 2133 19 EPIGENETIC INACTIVATION OF THE MIR-34A IN HEMATOLOGICAL MALIGNANCIES. MIR-34A IS A TRANSCRIPTIONAL TARGET OF P53 AND IMPLICATED IN CARCINOGENESIS. WE STUDIED THE ROLE OF MIR-34A METHYLATION IN A PANEL OF HEMATOLOGICAL MALIGNANCIES INCLUDING ACUTE LEUKEMIA [ACUTE MYELOID LEUKEMIA (AML) AND ACUTE LYMPHOBLASTIC LEUKEMIA (ALL)], CHRONIC LEUKEMIA [CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) AND CHRONIC MYELOID LEUKEMIA (CML)], MULTIPLE MYELOMA (MM) AND NON-HODGKIN'S LYMPHOMA (NHL). THE METHYLATION STATUS OF MIR-34A PROMOTER WAS STUDIED IN 12 CELL LINES AND 188 DIAGNOSTIC SAMPLES BY METHYLATION-SPECIFIC POLYMERASE CHAIN REACTION. MIR-34A PROMOTER WAS UNMETHYLATED IN NORMAL CONTROLS BUT METHYLATED IN 75% LYMPHOMA AND 37% MYELOMA CELL LINES. HYPOMETHYLATING TREATMENT LED TO RE-EXPRESSION OF PRI-MIR-34A TRANSCRIPT IN LYMPHOMA CELLS WITH HOMOZYGOUS MIR-34A METHYLATION. IN PRIMARY SAMPLES AT DIAGNOSIS, MIR-34A METHYLATION WAS DETECTED IN 4% CLL, 5.5% MM SAMPLES AND 18.8% OF NHL AT DIAGNOSIS BUT NONE OF ALL, AML AND CML (P = 0.011). IN MM PATIENTS WITH PAIRED SAMPLES, MIR-34A METHYLATION STATUS REMAINED UNCHANGED AT PROGRESSION. AMONGST LYMPHOID MALIGNANCIES, MIR-34A WAS PREFERENTIALLY METHYLATED IN NHL (P = 0.018), IN PARTICULAR NATURAL KILLER (NK)/T-CELL LYMPHOMA. IN CONCLUSION, AMONGST HEMATOLOGICAL MALIGNANCIES, MIR-34A METHYLATION IS PREFERENTIALLY HYPERMETHYLATED IN NHL, IN PARTICULAR NK/T-CELL LYMPHOMA, IN A TUMOR-SPECIFIC MANNER, THEREFORE THE ROLE OF MIR-34A IN LYMPHOMAGENESIS WARRANTS FURTHER STUDY. 2010 8 2438 23 EPIGENETIC SILENCING OF THE CIRCADIAN CLOCK GENE CRY1 IS ASSOCIATED WITH AN INDOLENT CLINICAL COURSE IN CHRONIC LYMPHOCYTIC LEUKEMIA. DISRUPTION OF CIRCADIAN RHYTHM IS BELIEVED TO PLAY A CRITICAL ROLE IN CANCER DEVELOPMENT. CRYPTOCHROME 1 (CRY1) IS A CORE COMPONENT OF THE MAMMALIAN CIRCADIAN CLOCK AND WE HAVE PREVIOUSLY SHOWN ITS DEREGULATED EXPRESSION IN A SUBGROUP OF PATIENTS WITH CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). USING REAL-TIME RT-PCR IN A COHORT OF 76 CLL PATIENTS AND 35 NORMAL BLOOD DONORS WE NOW DEMONSTRATE THAT DIFFERENTIAL CRY1 MRNA EXPRESSION IN HIGH-RISK (HR) CD38+/IMMUNOGLOBULIN VARIABLE HEAVY CHAIN GENE (IGVH) UNMUTATED PATIENTS AS COMPARED TO LOW-RISK (LR) CD38-/IGVH MUTATED PATIENTS CAN BE ATTRIBUTED TO DOWN-MODULATION OF CRY1 IN LR CLL CASES. ANALYSIS OF THE DNA METHYLATION PROFILE OF THE CRY1 PROMOTER IN A SUBGROUP OF 57 PATIENTS REVEALED THAT CRY1 EXPRESSION IN LR CLL CELLS IS SILENCED BY ABERRANT PROMOTER CPG ISLAND HYPERMETHYLATION. THE METHYLATION PATTERN OF THE CRY1 PROMOTER PROVED TO HAVE HIGH PROGNOSTIC IMPACT IN CLL WHERE ABERRANT PROMOTER METHYLATION PREDICTED A FAVOURABLE OUTCOME. CRY1 MRNA TRANSCRIPT LEVELS DID NOT CHANGE OVER TIME IN THE MAJORITY OF PATIENTS WHERE SEQUENTIAL SAMPLES WERE AVAILABLE FOR ANALYSIS. WE ALSO COMPARED THE CRY1 EXPRESSION IN CLL WITH OTHER LYMPHOID MALIGNANCIES AND OBSERVED EPIGENETIC SILENCING OF CRY1 IN A PATIENT WITH B CELL ACUTE LYMPHOBLASTIC LEUKEMIA (B-ALL). 2012 9 4601 36 NDRG2 MRNA LEVELS AND MIR-28-5P AND MIR-650 ACTIVITY IN CHRONIC LYMPHOCYTIC LEUKEMIA. BACKGROUND: NDRG2 IS IDENTIFIED AS A TUMOR SUPPRESSOR GENE IN MANY TUMORS, AND FUNCTIONS IN CELL PROLIFERATION, DIFFERENTIATION AND APOPTOSIS. RECENT DATA INDICATE THAT NDRG2 EXPRESSION IS UP-REGULATED BY TP53. MOREOVER, PROPOSED MECHANISMS OF NDRG2 INACTIVATION INCLUDE EPIGENETIC SILENCING OF THE NDRG2 PROMOTER AND DOWN-REGULATION BY MICRORNAS (MIRNAS). HOWEVER, FEW STUDIES HAVE EVER BEEN DONE ON THE ROLE OF NDRG2 AND THE NDRG2-REGULATING MIRNAS INTERFERENCE IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). METHODS: NDRG2 AND MICRORNAS MRNA LEVELS IN CLL SUBJECTS WERE ASSESSED BY QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION (QRT-PCR). THE DUAL-LUCIFERASE REPORTER ASSAY WAS PERFORMED TO DETERMINE NDRG2-RELATED MIRNAS. LOW EXPRESSION OF MATURE EXOGENOUS MIRNAS IN CLL CELLS WAS ESTABLISHED BY TRANSIENT TRANSFECTION. NDRG2 PROTEIN LEVELS IN CLL CELLS WERE DETECTED BY WESTERN BLOT. IN ADDITION, FLOW CYTOMETRY WAS CONDUCTED TO EXAMINE THE APOPTOSIS OF CLL CELLS. RESULTS: LOWER EXPRESSION OF NDRG2 WAS FOUND IN THE B-CELLS FROM 102 CLL PATIENTS COMPARED THE 40 NORMAL SUBJECTS (P < 0.001). PATIENTS WITH ADVANCED BINET STAGE (P = 0.001), HIGH LACTATE DEHYDROGENASE (LDH) LEVEL (P = 0.036), UN-MUTATED IMMUNOGLOBULIN HEAVY CHAIN VARIABLE REGION GENE (IGHV) (P = 0.004) AND THOSE WITH P53 ABERRATIONS (P < 0.001) HAD A MARKEDLY LOWER LEVELS OF NDRG2 MRNA. THIS DECREASE WAS ASSOCIATED WITH BRIEFER TIME-TO-TREATMENT (P = 0.001) AND POORER SURVIVAL (P < 0.001). HIGH EXPRESSION OF MIR-28-5P AND MIR-650 WAS ASSOCIATED WITH BINET B/C STAGE (P = 0.044) AND IGHV UN-MUTATED (P = 0.011), AS WELL AS BINET B/C STAGE (P = 0.013) AND P53 ABERRATIONS (P = 0.037), RESPECTIVELY. INHIBITION OF MIR-28-5P OR MIR-650 COULD INDUCE MORE APOPTOSIS IN CLL CELLS WITH GERMLINE TP53. CONCLUSIONS: NDRG2 MRNA LEVELS MIGHT BE A USEFUL PROGNOSTIC VARIABLE FOR PATIENTS OF CLL AND UP-REGULATING NDRG2 TRANSCRIPTION MAY BE A THERAPY APPROACH IN CLL WITHOUT P53 ABERRATIONS. 2018 10 6670 37 URINARY MICRORNA ANALYSIS INDICATES AN EPIGENETIC REGULATION OF CHRONIC KIDNEY DISEASE OF UNKNOWN ETIOLOGY IN SRI LANKA. BACKGROUND: CHRONIC KIDNEY DISEASE OF UNKNOWN ETIOLOGY (CKDU) IS REPORTED AMONG MALE PADDY FARMERS IN THE DRY ZONE OF SRI LANKA. THE EXACT CAUSE OF THIS DISEASE REMAINS UNDETERMINED. GENETIC SUSCEPTIBILITY IS IDENTIFIED AS A MAJOR RISK FACTOR FOR CKDU OBJECTIVES: IN THIS STUDY, SMALL URINARY RNAS WERE CHARACTERIZED IN CKDU PATIENTS, HEALTHY ENDEMIC AND NON-ENDEMIC CONTROLS. DIFFERENTLY EXPRESSED URINARY MIRNAS AND THEIR ASSOCIATED PATHWAYS WERE IDENTIFIED IN THE STUDY POPULATION. METHODS: HEALTHY AND DISEASED MALE VOLUNTEERS (N = 9) WERE RECRUITED FROM GIRANDURUKOTTE (ENDEMIC) AND MAWANELLA (NON-ENDEMIC) DISTRICTS. URINARY SMALL RNAS WERE PURIFIED AND SEQUENCED USING ILLUMINA MISEQTM. THE SEQUENCE TRACE FILES WERE ASSEMBLED AND ANALYZED. DIFFERENTIALLY EX-PRESSED MIRNAS AMONG THESE THREE GROUPS WERE IDENTIFIED AND PATHWAY ANALYSIS WAS CONDUCTED. RESULTS: THE URINE SAMPLES CONTAINED 130,623 SEQUENCE READS IDENTIFIED AS NON-CODING RNAS, PIWI-INTERACTING RNAS (PIRNA), AND MIRNAS. APPROXIMATELY FOUR PERCENT OF THE TOTAL SMALL RNA READS REPRESENTED MIRNA, AND 29% REPRESENTED PIRNA. A TOTAL OF 409 MIRNA SPECIES WERE EX-PRESSED IN URINE. INTERESTINGLY, BOTH DISEASED AND ENDEMIC CONTROLS POPULATION SHOWED SIGNIFICANTLY LOW EXPRESSION OF MIRNA AND PIRNA. REGARDLESS OF THE HEALTH STATUS, THE ENDEMIC POPULATION EX-PRESSED SIGNIFICANTLY LOW LEVELS OF MIR-10A, MIR-21, MIR-148A, AND MIR-30A WHICH HAVE BEEN LINKED WITH SEVERAL ENVIRONMENTAL TOXINS CONCLUSION: SIGNIFICANT DOWNREGULATION OF MIRNA AND PIRNA EXPRESSION IN BOTH DISEASED AND HEALTHY ENDEMIC SAMPLES INDICATES AN EPIGENETIC REGULATION OF CKDU INVOLVING GENETIC AND ENVIRONMENTAL INTERACTION. FURTHER STUDIES OF SPECIFIC MIRNA SPECIES ARE REQUIRED TO DEVELOP A MIRNA PANEL TO IDENTIFY INDIVIDUALS SUSCEPTIBLE TO CKDU. 2023 11 6238 37 THE MALIGNANCY SUPPRESSION ROLE OF MIR-23A BY TARGETING THE BCR/ABL ONCOGENE IN CHROMIC MYELOID LEUKEMIA. THE AIM OF THIS STUDY WAS TO INVESTIGATE THE ROLE AND MECHANISM OF MIR-23A IN THE REGULATION OF BCR/ABL AND TO PROVIDE A NEW PROGNOSTIC BIOMARKER FOR CHRONIC MYELOID LEUKEMIA (CML). THE EXPRESSION LEVELS OF MIR-23A AND BCR/ABL WERE ASSESSED IN 42 NEWLY DIAGNOSED CML PATIENTS, 37 CML PATIENTS IN FIRST COMPLETE REMISSION AND 25 HEALTHY CONTROLS. QUANTITATIVE REAL-TIME PCR, WESTERN BLOT ANALYSIS AND COLONY FORMATION ASSAY WERE USED TO EVALUATE CHANGES INDUCED BY OVEREXPRESSION OR INHIBITION OF MIR-23A OR BCR/ABL. MIR-23A MIMIC OR NEGATIVE CONTROL MIMIC WAS TRANSFECTED INTO A CML CELL LINE (K562) AND TWO LUNG CANCER CELL LINES (H157 AND SKMES1) USING LIPOFECTAMINE 2000, AND THE CELLS WERE USED FOR REAL-TIME REVERSE TRANSCRIPTION-PCR (RT-PCR) AND WESTERN BLOT ANALYSIS. WE FOUND THAT THE DOWNREGULATION OF MIR-23A EXPRESSION WAS A FREQUENT EVENT IN BOTH LEUKEMIA CELL LINES AND PRIMARY LEUKEMIC CELLS FROM PATIENTS WITH DE NOVO CML. THE MICROARRAY RESULTS SHOWED THAT MOST OF THE CML PATIENTS EXPRESSED HIGH LEVELS OF BCR/ABL AND LOW LEVELS OF MIR-23A. REAL-TIME RT-PCR AND WESTERN BLOT ANALYSIS SHOWED THAT THE BCR/ABL LEVELS IN MIR-23A-TRANSFECTED CELLS WERE LOWER THAN THOSE IN THE CONTROL GROUPS. ECTOPIC EXPRESSION OF MIR-23A IN K562 CELLS LED TO CELLULAR SENESCENCE. MOREOVER, WHEN K562 CELLS WERE TREATED WITH 5-AZA-2'-DEOXYCYTIDINE, A DNA METHYLATION INHIBITOR, BCR/ABL EXPRESSION WAS UPREGULATED, WHICH INDICATES EPIGENETIC SILENCING OF MIR-23A IN LEUKEMIC CELLS. BCR/ABL AND MIR-23A EXPRESSIONS WERE INVERSELY RELATED TO CML, AND BCR/ABL EXPRESSION WAS REGULATED BY MIR-23A IN LEUKEMIC CELLS. THE EPIGENETIC SILENCING OF MIR-23A LED TO DEREPRESSION OF BCR/ABL EXPRESSION, AND CONSEQUENTLY CONTRIBUTES TO CML DEVELOPMENT AND PROGRESSION. 2014 12 2131 19 EPIGENETIC INACTIVATION OF THE HSA-MIR-203 IN HAEMATOLOGICAL MALIGNANCIES. MIR-203 IS A TUMOUR SUPPRESSOR MICRORNA (MIRNA). WE STUDIED THE METHYLATION OF HSA-MIR-203 IN 150 SAMPLES INCLUDING ACUTE MYELOID LEUKAEMIA (AML), ACUTE LYMPHOBLASTIC LEUKAEMIA (ALL), CHRONIC MYELOID LEUKAEMIA (CML), CHRONIC LYMPHOCYTIC LEUKAEMIA (CLL) AND NON-HODGKIN'S LYMPHOMA (NHL) BY METHYLATION-SPECIFIC PCR, AND MIRNA EXPRESSION BY STEM-LOOP RT-QPCR. HSA-MIR-203 PROMOTER WAS UNMETHYLATED IN NORMAL CONTROLS BUT HOMOZYGOUSLY METHYLATED IN TWO AML AND FOUR LYMPHOMA CELL LINES, IN WHICH 5-AZA-2'-DEOXYCYTIDINE TREATMENT LED TO PROMOTER DEMETHYLATION AND MIR-203 RE-EXPRESSION. RESTORATION OF MIR-203 EXPRESSION IN LYMPHOMA CELLS INHIBITED CELLULAR PROLIFERATION AND INCREASED CELL DEATH, SUGGESTING AN INHERENT TUMOUR SUPPRESSOR ACTIVITY. IN PRIMARY SAMPLES, HSA-MIR-203 METHYLATION WAS ABSENT IN CML BUT DETECTED IN 5.0% ALL, 10.0% AML, 42.0% CLL AND 38.8% OF NHL (INCLUDING SIX [60.0%] NATURAL KILLER-CELL, NINE [40.9%] B-CELL AND FOUR [23.5%] T CELL NHL). MOREOVER, HSA-MIR-203 METHYLATION WAS ASSOCIATED WITH HYPERMETHYLATION OF HSA-MIR-34A, -124A AND -196B IN NHL BUT NOT CLL. IN CLL, HSA-MIR-203 METHYLATION WAS ASSOCIATED WITH A HIGHER PRESENTING HB LEVEL (P = 0.033). THE PROJECTED 10 YEAR OVERALL SURVIVAL OF THE CLL PATIENTS WAS 58.2%, WHICH WAS IMPACTED BY RAI STAGE AND HIGH-RISK KARYOTYPES BUT NOT HSA-MIR-203 METHYLATION. HSA-MIR-203 WAS MORE FREQUENTLY METHYLATED IN LYMPHOID THAN MYELOID MALIGNANCIES (P = 0.002). IN CONCLUSION, MIR-203, A TUMOUR SUPPRESSOR GENE, WAS HYPERMETHYLATED IN A TUMOUR-SPECIFIC MANNER WITH GENE SILENCING. HSA-MIR-203 WAS MORE FREQUENTLY HYPERMETHYLATED IN LYMPHOID THAN MYELOID MALIGNANCIES. IN NHL, HSA-MIR-203 METHYLATION WAS ASSOCIATED WITH CONCOMITANT METHYLATION OF OTHER TUMOUR SUPPRESSOR MIRNAS. THE FREQUENT HSA-MIR-203 METHYLATION IN LYMPHOID MALIGNANCIES SUGGESTED A PATHOGENETIC ROLE OF HSA-MIR-203 METHYLATION. 2011 13 5094 35 PLASMA EXTRACELLULAR VESICLE SUBTYPES MAY BE USEFUL AS POTENTIAL BIOMARKERS OF IMMUNE ACTIVATION IN PEOPLE WITH HIV. BACKGROUND: EXTRACELLULAR VESICLES (EVS) ARE INTERCELLULAR MESSENGERS WITH EPIGENETIC POTENTIAL SINCE THEY CAN SHUTTLE MICRORNA (MIRNA). EVS AND MIRNA PLAY A ROLE IN HUMAN IMMUNODEFICIENCY VIRUS (HIV) INFECTION IMMUNOPATHOGENESIS. CHRONIC IMMUNE ACTIVATION AND SYSTEMIC INFLAMMATION DURING HIV INFECTION DESPITE EFFECTIVE ANTIRETROVIRAL THERAPY (ART) ARE ASSOCIATED WITH NON-ACQUIRED IMMUNODEFICIENCY SYNDROME (AIDS) COMORBIDITIES IN PEOPLE LIVING WITH HIV (PLWH). ANALYSIS OF PLASMA EVS AND THEIR MIRNA CONTENT MAY BE USEFUL AS IMMUNE ACTIVATION OR INFLAMMATORY BIOMARKERS IN PLWH RECEIVING ART. IN THIS STUDY, WE HYPOTHESIZED THAT THE NUMBER, SIZE, AND MIRNA OF LARGE AND SMALL EVS COULD REFLECT IMMUNE ACTIVATION ASSOCIATED WITH AN ELEVATED CD8 T-CELL COUNT OR A LOW CD4/CD8 RATIO IN PLWH. METHODS: PLASMA EVS SUBTYPE PURIFIED FROM PLWH AND UNINFECTED CONTROLS WERE SIZED USING DYNAMIC LIGHT SCATTERING AND QUANTIFIED USING FLOW CYTOMETRY AND ACETYLCHOLINE ESTERASE (ACHE) ACTIVITY. EXPRESSION OF MATURE MIRNAS MIR-92, MIR-155, MIR-223 WAS MEASURED BY QUANTITATIVE REVERSE-TRANSCRIPTASE POLYMERASE CHAIN REACTION IN EVS AND LEUCOCYTES. RESULTS: HIV INFECTION INDUCES INCREASED PRODUCTION OF SMALL EVS IN PLASMA. EV SUBTYPES WERE DIFFERENTIALLY ENRICHED IN MIR-92, MIR-155, AND MIR-223. POSITIVE CORRELATIONS BETWEEN CD8 T-CELL COUNT AND LARGE EVS ABUNDANCE AND SMALL EVS ACHE ACTIVITY WERE OBSERVED. CD4/CD8 RATIO WAS NEGATIVELY CORRELATED WITH SMALL EV ACHE ACTIVITY, AND MIRNA-155 LEVEL PER SMALL EV WAS NEGATIVELY CORRELATED WITH CD8 T-CELL COUNT. CONCLUSIONS: THESE FINDINGS SUGGEST THAT QUANTIFYING LARGE OR SMALL EVS AND PROFILING MIRNA CONTENT PER EV MIGHT PROVIDE NEW FUNCTIONAL BIOMARKERS OF IMMUNE ACTIVATION AND INFLAMMATION. 2021 14 703 24 BTLA EXPRESSION IN CLL: EPIGENETIC REGULATION AND IMPACT ON CLL B CELL PROLIFERATION AND ABILITY TO IL-4 PRODUCTION. IN OUR PREVIOUS STUDY, WHILE CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) CASES SHOWED HIGHER LEVELS OF B AND T LYMPHOCYTE ATTENUATOR (BTLA) MRNA COMPARED TO CONTROLS, LOWER BTLA PROTEIN EXPRESSION WAS OBSERVED IN CASES COMPARED TO CONTROLS. HENCE WE HYPOTHESIZE THAT MICRO RNA (MIR) 155-5P REGULATES BTLA EXPRESSION IN CLL. IN LINE WITH EARLIER DATA, EXPRESSION OF BTLA MRNA AND MIR-155-5P IS ELEVATED IN CLL (P = 0.034 AND P = 0.0006, RESPECTIVELY) AS WELL AS IN MEC-1 CELL LINE (P = 0.009 AND 0.016, RESPECTIVELY). INHIBITION OF MIR-155-5P PARTIALLY RESTORED BTLA PROTEIN EXPRESSION IN CLL PATIENTS (P = 0.01) AND IN MEC-1 CELL LINES (P = 0.058). ADDITIONALLY, WE AIMED TO EVALUATE THE SIGNIFICANCE OF BTLA DEFICIENCY IN CLL CELLS ON PROLIFERATION AND IL-4 PRODUCTION OF B CELLS. WE FOUND THAT SECRETION OF IL-4 IS NOT DEPENDENT ON BTLA EXPRESSION, SINCE FRACTIONS OF BTLA POSITIVE AND BTLA NEGATIVE B CELLS EXPRESSING INTRACELLULAR IL-4 WERE SIMILAR IN CLL PATIENTS AND CONTROLS. WE DEMONSTRATED THAT IN CONTROLS THE FRACTION OF PROLIFERATING CELLS IS LOWER IN BTLA POSITIVE THAN IN BTLA NEGATIVE B CELLS (P = 0.059), WHICH WAS NOT OBSERVED IN CLL. HOWEVER, THE FREQUENCY OF BTLA POSITIVE KI67+ B CELLS IN CLL WAS HIGHER COMPARED TO CORRESPONDING CELLS FROM CONTROLS (P = 0.055) WHILE THERE WERE NO DIFFERENCES BETWEEN THE EXAMINED GROUPS REGARDING FREQUENCY OF BTLA NEGATIVE KI67+ B CELLS. OUR STUDIES SUGGEST THAT MIR-155-5P IS INVOLVED IN BTLA DEFICIENCY, AFFECTING PROLIFERATION OF CLL B CELLS, WHICH MAY BE ONE OF THE MECHANISMS RESPONSIBLE FOR CLL PATHOGENESIS. 2021 15 2132 20 EPIGENETIC INACTIVATION OF THE MIR-124-1 IN HAEMATOLOGICAL MALIGNANCIES. MIR-124-1 IS A TUMOUR SUPPRESSOR MICRORNA (MIR). EPIGENETIC DEREGULATION OF MIRS IS IMPLICATED IN CARCINOGENESIS. PROMOTER DNA METHYLATION AND HISTONE MODIFICATION OF MIR-124-1 WAS STUDIED IN 5 NORMAL MARROW CONTROLS, 4 LYMPHOMA, 8 MULTIPLE MYELOMA (MM) CELL LINES, 230 DIAGNOSTIC PRIMARY SAMPLES OF ACUTE MYELOID LEUKAEMIA (AML), ACUTE LYMPHOBLASTIC LEUKAEMIA (ALL), CHRONIC MYELOID LEUKAEMIA (CML), CHRONIC LYMPHOCYTIC LEUKAEMIA (CLL), MM, AND NON-HODGKIN'S LYMPHOMA (NHL), AND 53 MM SAMPLES AT STABLE DISEASE OR RELAPSE. PROMOTER OF MIR-124-1 WAS UNMETHYLATED IN NORMAL CONTROLS BUT HOMOZYGOUSLY METHYLATED IN 4 OF 4 LYMPHOMA AND 4 OF 8 MYELOMA CELL LINES. TREATMENT OF 5-AZA-2'-DEOXYCYTIDINE LED TO MIR-124-1 DEMETHYLATION AND RE-EXPRESSION OF MATURE MIR-124, WHICH ALSO ASSOCIATED WITH EMERGENCE OF EUCHROMATIC TRIMETHYL H3K4 AND CONSEQUENT DOWNREGULATION OF CDK6 IN MYELOMA CELLS HARBORING HOMOZYGOUS MIR-124-1 METHYLATION. IN PRIMARY SAMPLES AT DIAGNOSIS, MIR-124-1 METHYLATION WAS ABSENT IN CML BUT DETECTED IN 2% EACH OF MM AT DIAGNOSIS AND RELAPSE/PROGRESSION, 5% ALL, 15% AML, 14% CLL AND 58.1% OF NHL (P<0.001). AMONGST LYMPHOID MALIGNANCIES, MIR-124-1 WAS PREFERENTIALLY METHYLATED IN NHL THAN MM, CLL OR ALL. IN PRIMARY LYMPHOMA SAMPLES, MIR-124-1 WAS PREFERENTIALLY HYPERMETHYLATED IN B- OR NK/T-CELL LYMPHOMAS AND ASSOCIATED WITH REDUCED MIR-124 EXPRESSION. IN CONCLUSION, MIR-124-1 WAS HYPERMETHYLATED IN A TUMOUR-SPECIFIC MANNER, WITH A HETEROCHROMATIC HISTONE CONFIGURATION. HYPOMETHYLATION LED TO PARTIAL RESTORATION OF EUCHROMATIC HISTONE CODE AND MIR RE-EXPRESSION. INFREQUENT MIR-124-1 METHYLATION DETECTED IN DIAGNOSTIC AND RELAPSE MM SAMPLES SHOWED AN UNIMPORTANT ROLE IN MM PATHOGENESIS, DESPITE FREQUENT METHYLATION FOUND IN CELL LINES. AMONGST HAEMATOLOGICAL CANCERS, MIR-124-1 WAS MORE FREQUENTLY HYPERMETHYLATED IN NHL, AND HENCE WARRANTS FURTHER STUDY. 2011 16 4349 25 MIR-155 AND MIR-122 EXPRESSION OF SPERMATOZOA IN OBESE SUBJECTS. OBESITY IS CHARACTERIZED BY MILD CHRONIC INFLAMMATION THAT IS LINKED WITH IMPAIRED IRON HOMEOSTASIS. STUDIES IN HUMAN AND MURINE SHOW THAT THERE IS A TRANSGENERATIONAL EPIGENETIC INHERITANCE VIA THE GAMETES IN OBESITY; HOWEVER, THERE IS LITTLE INFORMATION ON CHANGES IN THE EXPRESSION OF MICRORNAS RELATED TO INFLAMMATION AND IRON HOMEOSTASIS IN SPERMATOZOA FROM OBESE SUBJECTS. THE PRESENT STUDY INVESTIGATED THE EXPRESSION OF MICRORNAS RELATED TO INFLAMMATION (MIR-21 Y MIR-155) AND IRON NUTRITION (MIR-122 AND MIR-200B) IN PLASMA, PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMC) AND SPERMATOZOA FROM NORMOZOOSPERMIC CONTROLS (CN; N = 17; BMI: 24.6 +/- 2.0) AND OBESE (OB; N = 17; BMI: 32.6 +/- 4.4) MEN. TO DETERMINE THE INFLAMMATION LEVELS, WE MEASURED IL-6, TNF-ALPHA, AND MONOCYTE CHEMOATTRACTANT PROTEIN-1 (MCP1) BY MAGNETIC LUMINEX((R)) ASSAY. MRNA EXPRESSION OF IL6, TNF-ALPHA, AND HEPCIDIN (HAMP) IN PBMC WERE EVALUATED BY RT-QPCR. THE ANALYSIS OF MICRORNAS WAS PERFORMED USING THE TAQMAN((R)) ASSAYS. THE IRON CONTENT IN PBMC, SEMINAL PLASMA, AND SPERMATOZOA WAS DETERMINED BY INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY (ICP-MS). HIGH SERUM IL6, TNF-ALPHA, AND MCP1 LEVELS WERE OBSERVED IN OB GROUP (P < 0.05). GENE EXPRESSION ANALYSIS SHOWED AN INCREASED ABUNDANCE RELATIVE OF TNF-ALPHA (P = 0.018), HAMP (P = 0.03), AND IL6 (P = 0.02) IN PBMC FROM OBESE SUBJECTS. ALSO, WE OBSERVED HIGH LEVELS OF SERUM FERRITIN (P = 0.03), IRON CONTENT IN SEMINAL PLASMA (P = 0.04), AND SPERMATOZOA (P = 0.002), BUT LOWER SERUM FE (P = 0.007) IN OBESE SUBJECTS. IN THE OB GROUP, A HIGH EXPRESSION OF MIR-155 (P = 0.02) AND MIR-21 (P = 0.03) WAS OBSERVED IN PBMC AND MIR-122 (P = 0.03) IN PLASMA. IN SPERM, BOTH MIR-155 (P = 0.004) AND MIR-122 (P = 0.028) WERE HIGH IN THE OB GROUP. OUR RESULTS SHOWED THAT OBESE SUBJECTS HAVE INCREASED EXPRESSIONS OF MIR-155 AND MIR-122, TWO MICRORNAS THAT WERE PREVIOUSLY RELATED WITH INFLAMMATION AND IRON METABOLISM, RESPECTIVELY, AT BOTH THE SYSTEMIC AND SPERM LEVELS. 2018 17 3947 27 LNCRNA UCA1 INDUCES AUTOPHAGIC GENE EXPRESSION VIA EPIGENETIC REGULATION MEDIATED BY ATG16L1 AND MIR-132-3P IN SH-SY5Y CELLS TREATED WITH RETINOIC ACID. OBJECTIVE: EPILEPSY IS A CHRONIC BRAIN DISEASE WITH RECURRENT SEIZURES. AUTOPHAGY PLAYS A CRUCIAL ROLE IN THE PROGRESSION OF EPILEPSY. THIS STUDY AIMED TO EXPLORE THE FUNCTION AND INTRINSIC MECHANISM OF THE LONG NON-CODING RNA (LNCRNA) UCA1/MIR-132-3P/ATG16L1 AXIS IN EPILEPSY VIA REGULATION OF AUTOPHAGY. METHODS: THE EXPRESSION OF LNCRNA UCA1, MIR-132-3P AND ATG16L1 WAS MEASURED IN SERUM FROM EPILEPTIC PATIENTS BY QUANTITATIVE RT-PCR. A SH-SY5Y CELL MODEL WAS FURTHER CONSTRUCTED USING RETINOIC ACID TO INVESTIGATE THE UCA1/ MIR-132-3P/ATG16L1 AXIS BY QUANTITATIVE RT-PCR, WESTERN BLOTTING, FLUORESCENCE IN SITU HYBRIDISATION, RNA IMMUNOPRECIPITATION, CHROMATIN IMMUNOPRECIPITATION, AND A DUAL-LUCIFERASE REPORTER GENE ASSAY. RESULTS: IN THE SERUM OF EPILEPTIC PATIENTS, THE LEVEL OF LNCRNA UCA1 AND ATG16L1 WAS REDUCED AND MIR-132-3P ELEVATED, COMPARED TO CONTROLS. SIMILARLY, IN THE SH-SY5Y CELL MODEL, THE LEVEL OF LNCRNA UCA1 AND ATG16L1 WAS REDUCED AND MIR-132-3P ELEVATED IN RETINOIC ACID-TREATED CELLS; LNCRNA UCA1 WAS MAINLY LOCATED IN THE CYTOPLASM. LNCRNA UCA1 OVEREXPRESSION WAS SHOWN TO PROMOTE AUTOPHAGIC GENE EXPRESSION, WHICH WAS REVERSED BY MIR-132-3P OVEREXPRESSION. MOREOVER, AUTOPHAGIC GENE EXPRESSION INDUCED BY MIR-132-3P KNOCKDOWN WAS REVERSED BY ATG16L1 KNOCKDOWN. BASED ON PRECIPITATION ASSAYS, LNCRNA UCA1 AND MIR-132-3P WERE SHOWN TO FORM A COMPLEX WITH THE TRANSCRIPTION FACTOR, EZH2, AND MIR-132-3P WAS SHOWN TO INTERACT WITH ATG16L1 BASED ON A LUCIFERASE ASSAY. FINALLY, LNCRNA UCA1 WAS SHOWN TO NEGATIVELY REGULATE MIR-132-3P EXPRESSION, AND MIR-132-3P WAS SHOWN TO NEGATIVELY REGULATE ATG16L1. SIGNIFICANCE: IN THIS CELL MODEL, LNCRNA UCA1 PROMOTES AUTOPHAGIC GENE EXPRESSION VIA EPIGENETIC REGULATION MEDIATED BY ATG16L1 AND MIR-132-3P. 2022 18 5120 29 POSSIBLE EPIGENETIC REGULATORY EFFECT OF DYSREGULATED CIRCULAR RNAS IN EPILEPSY. CIRCULAR RNAS (CIRCRNAS) INVOLVE IN THE EPIGENETIC REGULATION AND ITS MAJOR MECHANISM IS THE SEQUESTRATION OF THE TARGET MICRO RNAS (MIRNAS). WE HYPOTHESIZED THAT CIRCRNAS MIGHT BE RELATED WITH THE PATHOPHYSIOLOGY OF CHRONIC EPILEPSY AND EVALUATED THE ALTERED CIRCRNA EXPRESSIONS AND THEIR POSSIBLE REGULATORY EFFECTS ON THEIR TARGET MIRNAS AND MRNAS IN A MOUSE EPILEPSY MODEL. THE CIRCRNA EXPRESSION PROFILE IN THE HIPPOCAMPUS OF THE PILOCARPINE MICE WAS ANALYZED AND COMPARED WITH CONTROL. THE CORRELATION BETWEEN THE EXPRESSION OF MIRNA BINDING SITES (MIRNA RESPONSE ELEMENTS, MRE) IN THE DYSREGULATED CIRCRNAS AND THE EXPRESSION OF THEIR TARGET MIRNAS WAS EVALUATED. AS MIRNAS ALSO INHIBIT THEIR TARGET MRNAS, CIRCRNA-MIRNA-MRNA REGULATORY NETWORK, COMPRISED OF DYSREGULATED RNAS THAT TARGETS ONE ANOTHER WERE SEARCHED. FOR THE IDENTIFIED NETWORKS, BIOINFORMATICS ANALYSES WERE PERFORMED. AS THE RESULT, FORTY-THREE CIRCRNAS WERE DYSREGULATED IN THE HIPPOCAMPUS (UP-REGULATED, 26; DOWN-REGULATED, 17). THE CHANGE IN THE EXPRESSION OF MRE IN THOSE CIRCRNAS NEGATIVELY CORRELATED WITH THE CHANGE IN THE RELEVANT TARGET MIRNA EXPRESSION (R = -0.461, P<0.001), SUPPORTING THAT CIRCRNAS INHIBIT THEIR TARGET MIRNA. 333 DYSREGULATED CIRCRNA-MIRNA-MRNA NETWORKS WERE IDENTIFIED. GENE ONTOLOGY AND PATHWAY ANALYSES DEMONSTRATED THAT THE UP-REGULATED MRNAS IN THOSE NETWORKS WERE CLOSELY RELATED TO THE MAJOR PROCESSES IN EPILEPSY. AMONG THEM, STRING ANALYSIS IDENTIFIED 37 KEY MRNAS WITH ABUNDANT (>/=4) INTERACTIONS WITH OTHER DYSREGULATED TARGET MRNAS. THE DYSREGULATION OF THE CIRCRNAS WHICH HAD MULTIPLE INTERACTIONS WITH KEY MRNAS WERE VALIDATED BY PCR. WE CONCLUDED THAT DYSREGULATED CIRCRNAS MIGHT HAVE A PATHOPHYSIOLOGIC ROLE IN CHRONIC EPILEPSY BY REGULATING MULTIPLE DISEASE RELEVANT MRNAS VIA CIRCRNA-MIRNA-MRNA INTERACTIONS. 2018 19 353 28 ALTERED LEVELS OF IMMUNE-REGULATORY MICRORNAS IN PLASMA SAMPLES OF PATIENTS WITH LUPUS NEPHRITIS. INTRODUCTION: LUPUS NEPHRITIS (LN) IS A MAJOR CAUSE OF MORTALITY AND MORBIDITY IN THE PATIENTS WITH LUPUS, A CHRONIC AUTOIMMUNE DISEASE. THE ROLE OF GENETIC AND EPIGENETIC FACTORS IS EMPHASIZED IN THE PATHOGENESIS OF LN. THE AIM OF THE PRESENT STUDY WAS TO EVALUATE THE LEVELS OF IMMUNE-REGULATORY MICRORNAS (E.G., MIR-31, MIR-125A, MIR-142-3P, MIR-146A, AND MIR-155) IN PLASMA SAMPLES OF PATIENTS WITH LN. METHODS: IN THIS STUDY, 26 PATIENTS WITH LN AND 26 HEALTHY INDIVIDUALS WERE INCLUDED. THE PLASMA LEVELS OF THE MICRORNAS WERE EVALUATED BY A QUANTITATIVE REAL-TIME PCR. MOREOVER, THE CORRELATION OF CIRCULATING PLASMA MICRORNAS WITH DISEASE ACTIVITY AND PATHOLOGICAL FINDINGS ALONG WITH THEIR ABILITY TO DISTINGUISH PATIENTS WITH LN WERE ASSESSED. RESULTS: PLASMA LEVELS OF MIR-125A (P = 0.048), MIR-146A (P = 0.005), AND MIR-155 (P< 0.001) WERE SIGNIFICANTLY HIGHER IN COMPARISON BETWEEN THE CASES AND CONTROLS. THE PLASMA LEVEL OF MIR-146A SIGNIFICANTLY CORRELATED WITH THE LEVEL OF ANTI-DOUBLE STRAND-DNA ANTIBODY AND PROTEINURIA. MOREOVER, THERE WAS A SIGNIFICANT CORRELATION BETWEEN MIR-142-3P LEVELS AND DISEASE CHRONICITY AND ACTIVITY INDEX (P <0.05). THE MULTIVARIATE ROC CURVE ANALYSIS INDICATED THE PLASMA CIRCULATING MIR-125A, MIR-142-3P, MIR-146, AND MIR-155 TOGETHER COULD DISCRIMINATE MOST OF THE PATIENTS WITH LN FROM CONTROLS WITH AREA AN UNDER CURVE (AUC) OF 0.89 [95% CI, 0.80-0.98, P<0.001], 88% SENSITIVITY, AND 78% SPECIFICITY. CONCLUSION: BASED ON THE FINDINGS OF THE PRESENT STUDY, THE STUDIED MICRORNAS MAY BE INVOLVED IN THE PATHOGENESIS AND DEVELOPMENT OF LN AND HAVE THE POTENTIAL TO BE USED AS DIAGNOSTIC AND THERAPEUTIC MARKERS IN LN. 2018 20 2749 27 EXPRESSION AND POLYMORPHISM OF MICRO-RNA ACCORDING TO BODY MASS INDEX AND BREAST CANCER PRESENTATION IN TUNISIAN PATIENTS. MICRO-RNAS (MIRS) CONSTITUTE A CLASS OF SMALL NONCODING RNAS IMPLICATED IN THE REGULATION OF GENE EXPRESSION BY BINDING TO TARGET MRNAS. A MIR CAN TARGET SEVERAL MRNAS, BEING INVOLVED IN DIFFERENT BIOLOGIC PROCESSES AND PATHOLOGIES. THIS PLEIOTROPIC FUNCTION MIGHT EXPLAIN THE LINK BETWEEN DISEASES CO-OCCURRENCE. EPIGENETIC ORIGIN OF THE LINK BETWEEN OBESITY AND BREAST CANCER (BC) IS INVESTIGATED IN A COHORT OF TUNISIAN PATIENTS, FOCUSING ON POLYMORPHISM AT GERMLINE LEVEL (MIR-146A) AND ON EXPRESSION IN MAMMARY TUMORS (MIR-21, MIR-146A, AND MIR-34A), ACCORDING TO BODY MASS INDEX (BMI) AND CLINICO-PATHOLOGIC FEATURES. THE MEASURE OF MIR EXPRESSION IN 60 MAMMARY TUMORS WAS REALIZED USING QUANTITATIVE RT-PCR. STUDY OF RS 2910164 IN MIR-146A WAS PERFORMED BY PCR AND DIRECT SEQUENCING USING BLOOD DNA OF 83 AFFECTED WOMEN AND 50 UNRELATED SUBJECTS FROM GREAT TUNIS. MIR-21, MIR-146A, AND MIR-34A HAVE BEEN QUANTIFIED IN BREAST TUMOR ACCORDING TO BMI. MIR-21 IS SIGNIFICANTLY MORE EXPRESSED IN TUMORS OF OBESE WOMEN COMPARATIVELY TO NONOBESE PATIENTS. ON THE CONTRARY, MIR-34A IS DECREASED IN TUMORS OF OBESE WOMEN. MOREOVER, IN OBESE BC PATIENTS, A SIGNIFICANT INCREASE IN BOTH MIR-21 AND MIR-146A EXPRESSION IS REVEALED IN CASES WITH LYMPH NODE METASTASIS. THE POLYMORPHISM AT RS 2910164 (MIR-146A) LOCUS WAS NOT SHOWN AS A RISK FACTOR FOR BC. HOWEVER THE MUTANT CC GENOTYPE WAS REVEALED TO BE ASSOCIATED WITH A RISK FOR BAD OUTCOME OF THE DISEASE. CHRONIC INFLAMMATION IN OBESE WOMEN WOULD BE LINKED TO AGGRESSIVE BREAST TUMORS VIA INDUCTION OF ONCOMIRS OVEREXPRESSION AND DECREASE OF TUMOR SUPPRESSOR MIRS. 2019