title: The IL-4/STAT6 signaling axis establishes a conserved microRNA signature in human and mouse macrophages regulating cell survival via miR-342-3p.
abstract: IL-4-driven alternative macrophage activation and proliferation are characteristic features of both antihelminthic immune responses and wound healing in contrast to classical macrophage activation, which primarily occurs during inflammatory responses. The signaling pathways defining the genome-wide microRNA expression profile as well as the cellular functions controlled by microRNAs during alternative macrophage activation are largely unknown. Hence, in the current work we examined the regulation and function of IL-4-regulated microRNAs in human and mouse alternative macrophage activation.
  
  
  
  We utilized microarray-based microRNA profiling to detect the dynamic expression changes during human monocyte-macrophage differentiation and IL-4-mediated alternative macrophage activation. The expression changes and upstream regulatory pathways of selected microRNAs were further investigated in human and mouse in vitro and in vivo models of alternative macrophage activation by integrating small RNA-seq, ChIP-seq, ChIP-quantitative PCR, and gene expression data. MicroRNA-controlled gene networks and corresponding functions were identified using a combination of transcriptomic, bioinformatic, and functional approaches.
  
  
  
  The IL-4-controlled microRNA expression pattern was identified in models of human and mouse alternative macrophage activation. IL-4-dependent induction of miR-342-3p and repression of miR-99b along with miR-125a-5p occurred in both human and murine macrophages in vitro. In addition, a similar expression pattern was observed in peritoneal macrophages of Brugia malayi nematode-implanted mice in vivo. By using IL4Rα- and STAT6-deficient macrophages, we were able to show that IL-4-dependent regulation of miR-342-3p, miR-99b, and miR-125a-5p is mediated by the IL-4Rα-STAT6 signaling pathway. The combination of gene expression studies and chromatin immunoprecipitation experiments demonstrated that both miR-342-3p and its host gene, EVL, are coregulated directly by STAT6. Finally, we found that miR-342-3p is capable of controlling macrophage survival through targeting an anti-apoptotic gene network including Bcl2l1.
  
  
  
  Our findings identify a conserved IL-4/STAT6-regulated microRNA signature in alternatively activated human and mouse macrophages. Moreover, our study indicates that miR-342-3p likely plays a pro-apoptotic role in such cells, thereby providing a negative feedback arm to IL-4-dependent macrophage proliferation.
authors.affiliation: Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, University of Debrecen Medical, Nagyerdei krt. 98, H-4032, Debrecen, Hungary.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Czimmerer
authors.affiliation: Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, University of Debrecen Medical, Nagyerdei krt. 98, H-4032, Debrecen, Hungary.
authors.forename: Tamas
authors.initials: T
authors.lastname: Varga
authors.affiliation: Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, University of Debrecen Medical, Nagyerdei krt. 98, H-4032, Debrecen, Hungary.
authors.forename: Mate
authors.initials: M
authors.lastname: Kiss
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad (Langebio), Centro de Investigación y de Estudios Avanzados del IPN, Irapuato, Guanajuato, 36821, México.
authors.forename: Cesaré Ovando
authors.initials: CO
authors.lastname: Vázquez
authors.affiliation: Department of Biophysics and Cell Biology, University of Debrecen, Egyetem tér 1, H-4012, Debrecen, Hungary.
authors.forename: Quang Minh
authors.initials: QM
authors.lastname: Doan-Xuan
authors.affiliation: University of Manchester, AV Hill Building, Oxford Road, Manchester, M13 9PT, UK.
authors.forename: Dominik
authors.initials: D
authors.lastname: Rückerl
authors.affiliation: Max Delbrueck Center for Molecular Medicine, Robert Roessle Strasse 10, Berlin, 13125, Germany.
authors.forename: Sudhir Gopal
authors.initials: SG
authors.lastname: Tattikota
authors.affiliation: Max Delbrueck Center for Molecular Medicine, Robert Roessle Strasse 10, Berlin, 13125, Germany.
authors.forename: Xin
authors.initials: X
authors.lastname: Yan
authors.affiliation: Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, University of Debrecen Medical, Nagyerdei krt. 98, H-4032, Debrecen, Hungary.
authors.forename: Zsuzsanna S
authors.initials: ZS
authors.lastname: Nagy
authors.affiliation: Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, University of Debrecen Medical, Nagyerdei krt. 98, H-4032, Debrecen, Hungary.|Sanford-Burnham-Prebys Medical Discovery Institute, 6400 Sanger Road, Orlando, FL, 32827, USA.
authors.forename: Bence
authors.initials: B
authors.lastname: Daniel
authors.affiliation: Genomic Medicine and Bioinformatic Core Facility, Department of Biochemistry and Molecular Biology, University of Debrecen, Nagyerdei krt. 98, H-4032, Debrecen, Hungary.
authors.forename: Szilard
authors.initials: S
authors.lastname: Poliska
authors.affiliation: Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, University of Debrecen Medical, Nagyerdei krt. 98, H-4032, Debrecen, Hungary.
authors.forename: Attila
authors.initials: A
authors.lastname: Horvath
authors.affiliation: Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, University of Debrecen Medical, Nagyerdei krt. 98, H-4032, Debrecen, Hungary.
authors.forename: Gergely
authors.initials: G
authors.lastname: Nagy
authors.affiliation: National Agricultural Research and Innovation Centre, Agricultural Biotechnology Institute, Szent-Györgyi A. út 4, H-2100, Gödöllő, Hungary.
authors.forename: Eva
authors.initials: E
authors.lastname: Varallyay
authors.affiliation: Max Delbrueck Center for Molecular Medicine, Robert Roessle Strasse 10, Berlin, 13125, Germany.
authors.forename: Matthew N
authors.initials: MN
authors.lastname: Poy
authors.affiliation: University of Manchester, AV Hill Building, Oxford Road, Manchester, M13 9PT, UK.
authors.forename: Judith E
authors.initials: JE
authors.lastname: Allen
authors.affiliation: Department of Biophysics and Cell Biology, University of Debrecen, Egyetem tér 1, H-4012, Debrecen, Hungary.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Bacso
authors.affiliation: Laboratorio Nacional de Genómica para la Biodiversidad (Langebio), Centro de Investigación y de Estudios Avanzados del IPN, Irapuato, Guanajuato, 36821, México.
authors.forename: Cei
authors.initials: C
authors.lastname: Abreu-Goodger
authors.affiliation: Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, University of Debrecen Medical, Nagyerdei krt. 98, H-4032, Debrecen, Hungary. lnagy@sbpdiscovery.org.|Sanford-Burnham-Prebys Medical Discovery Institute, 6400 Sanger Road, Orlando, FL, 32827, USA. lnagy@sbpdiscovery.org.|MTA-DE "Lendület" Immunogenomics Research Group, University of Debrecen, Egyetem tér 1, H-4012, Debrecen, Hungary. lnagy@sbpdiscovery.org.
authors.forename: Laszlo
authors.initials: L
authors.lastname: Nagy
mesh_terms: D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D002470:Cell Survival; D002478:Cells, Cultured; D017124:Conserved Sequence; D006801:Humans; D015847:Interleukin-4 / Q000276:immunology* / Q000378:metabolism; D008262:Macrophage Activation; D008264:Macrophages / Q000166:cytology* / Q000276:immunology*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D050800:STAT6 Transcription Factor / Q000235:genetics; D017423:Sequence Analysis, RNA / Q000379:methods; D015398:Signal Transduction
chemical_list: C546176:MIRN342 microRNA, human; D035683:MicroRNAs; C581863:Mirn342 microRNA, mouse; D050800:STAT6 Transcription Factor; D015847:Interleukin-4