title: Differential Expression of miR-4520a Associated With Pyrin Mutations in Familial Mediterranean Fever (FMF).
abstract: Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by recurrent, acute, and self-limiting attacks of fever. Mutations in MEFV gene encoding pyrin account for FMF, but the high number of heterozygote patients with typical symptoms of the disease has driven a number of alternative aetiopathogenic hypotheses. The MEFV gene was knocked down in human myelomonocytic cells that express endogenous pyrin to identify deregulated microRNAs (miRNAs). Microarray analyses revealed 29 significantly differentially expressed miRNAs implicated in pathways associated with cellular integrity and survival. Implementation of in silico gene network prediction algorithms and bioinformatics analyses showed that miR-4520a is predicted to target genes implicated in autophagy through regulation of RHEB/mTOR signaling. Differential expression levels of RHEB were confirmed by luciferase reporter gene assays providing further evidence that is directly targeted by miR-4520a. Although the relative expression levels of miR-4520a were variable among FMF patients, the statistical expression of miR-4520a was different between FMF mutation carriers and controls (P = 0.0061), indicating an association between miR-4520a expression and MEFV mutations. Comparison between FMF patients bearing the M694V mutation, associated with severe disease, and healthy controls showed a significant increase in miR-4520a expression levels (P = 0.00545). These data suggest that RHEB, the main activator of mTOR signaling, is a valid target of miR-4520a with the relative expression levels of the latter being significantly deregulated in FMF patients and highly dependent on the presence of pyrin mutations, especially of the M694V type. These results suggest a role of deregulated autophagy in the pathogenesis of FMF. J. Cell. Physiol. 232: 1326-1336, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Molecular Medicine and Human Genetics Laboratory, School of Medicine, University of Crete, Heraklion, Greece.|Institute of Molecular Biology and Biotechnology, FORTH, Heraklion, Crete, Greece.
authors.forename: Helen
authors.initials: H
authors.lastname: Latsoudis
authors.affiliation: German Rheumatism Research Center (DRFZ) Berlin, Leibniz Institute, Berlin, Germany.
authors.forename: Mir-Farzin
authors.initials: MF
authors.lastname: Mashreghi
authors.affiliation: Department of Bioinformatics, German Rheumatism Research Center (DRFZ), Leibniz Institute, Berlin, Germany.
authors.forename: Joachim R
authors.initials: JR
authors.lastname: Grün
authors.affiliation: German Rheumatism Research Center (DRFZ) Berlin, Leibniz Institute, Berlin, Germany.
authors.forename: Hyun-Dong
authors.initials: HD
authors.lastname: Chang
authors.affiliation: Department of Rheumatology and Clinical Immunology, Charité University Hospital, Berlin, Germany.
authors.forename: Bruno
authors.initials: B
authors.lastname: Stuhlmüller
authors.affiliation: Clinic of Rheumatology, University Hospital of Heraklion, Crete, Greece.
authors.forename: Argyro
authors.initials: A
authors.lastname: Repa
authors.affiliation: Clinic of Rheumatology, University Hospital of Heraklion, Crete, Greece.
authors.forename: Irini
authors.initials: I
authors.lastname: Gergiannaki
authors.affiliation: Clinic of Rheumatology, University Hospital of Heraklion, Crete, Greece.
authors.forename: Eleni
authors.initials: E
authors.lastname: Kabouraki
authors.affiliation: Novartis (Hellas) S.A.C.I., Athens, Greece.
authors.forename: George S
authors.initials: GS
authors.lastname: Vlachos
authors.affiliation: German Rheumatism Research Center (DRFZ) Berlin, Leibniz Institute, Berlin, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Häupl
authors.affiliation: German Rheumatism Research Center (DRFZ) Berlin, Leibniz Institute, Berlin, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Radbruch
authors.affiliation: Clinic of Rheumatology, University Hospital of Heraklion, Crete, Greece.
authors.forename: Prodromos
authors.initials: P
authors.lastname: Sidiropoulos
authors.affiliation: Novartis (Hellas) S.A.C.I., Athens, Greece.
authors.forename: Kimon
authors.initials: K
authors.lastname: Doukoumetzidis
authors.affiliation: Institute of Molecular Biology and Biotechnology, FORTH, Heraklion, Crete, Greece.|Department of Biochemistry, School of Medicine, University of Crete, Heraklion, Greece.
authors.forename: Dimitris
authors.initials: D
authors.lastname: Kardassis
authors.affiliation: Division of Rheumatology, Department of Immunology, Mayo Clinic, Rochester, Minnesota.
authors.forename: Timothy B
authors.initials: TB
authors.lastname: Niewold
authors.affiliation: Institute of Molecular Biology and Biotechnology, FORTH, Heraklion, Crete, Greece.|Biomedical Research Foundation of the Academy of Athens, Athens, Greece.|4th Department of Medicine, Rheumatology and Clinical Immunology, National and Kapodistrian University, Athens, Greece.
authors.forename: Dimitrios T
authors.initials: DT
authors.lastname: Boumpas
authors.affiliation: Molecular Medicine and Human Genetics Laboratory, School of Medicine, University of Crete, Heraklion, Greece.
authors.forename: George N
authors.initials: GN
authors.lastname: Goulielmos
mesh_terms: D000328:Adult; D016022:Case-Control Studies; D002460:Cell Line; D010505:Familial Mediterranean Fever / Q000235:genetics*; D005260:Female; D020869:Gene Expression Profiling; D053263:Gene Regulatory Networks; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008297:Male; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D020559:Monomeric GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism; D000071198:Pyrin / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D000076205:Ras Homolog Enriched in Brain Protein
chemical_list: D035683:MicroRNAs; D009479:Neuropeptides; D000071198:Pyrin; C490211:RHEB protein, human; D012333:RNA, Messenger; D000076205:Ras Homolog Enriched in Brain Protein; D008156:Luciferases; D020559:Monomeric GTP-Binding Proteins