title: Notch-regulated miR-223 targets the aryl hydrocarbon receptor pathway and increases cytokine production in macrophages from rheumatoid arthritis patients.
abstract: Evidence links aryl hydrocarbon receptor (AHR) activation to rheumatoid arthritis (RA) pathogenesis, although results are inconsistent. AHR agonists inhibit pro-inflammatory cytokine expression in macrophages, pivotal cells in RA aetiopathogenesis, which hints at specific circuits that regulate the AHR pathway in RA macrophages. We compared microRNA (miR) expression in CD14(+) cells from patients with active RA or with osteoarthritis (OA). Seven miR were downregulated and one (miR-223) upregulated in RA compared to OA cells. miR-223 upregulation correlated with reduced Notch3 and Notch effector expression in RA patients. Overexpression of the Notch-induced repressor HEY-1 and co-culture of healthy donor monocytes with Notch ligand-expressing cells showed direct Notch-mediated downregulation of miR-223. Bioinformatics predicted the AHR regulator ARNT (AHR nuclear translocator) as a miR-223 target. Pre-miR-223 overexpression silenced ARNT 3'UTR-driven reporter expression, reduced ARNT (but not AHR) protein levels and prevented AHR/ARNT-mediated inhibition of pro-inflammatory cytokine expression. miR-223 counteracted AHR/ARNT-induced Notch3 upregulation in monocytes. Levels of ARNT and of CYP1B1, an AHR/ARNT signalling effector, were reduced in RA compared to OA synovial tissue, which correlated with miR-223 levels. Our results associate Notch signalling to miR-223 downregulation in RA macrophages, and identify miR-223 as a negative regulator of the AHR/ARNT pathway through ARNT targeting.
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología/CSIC, Madrid.
authors.forename: Jesús
authors.initials: J
authors.lastname: Ogando
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología/CSIC, Madrid.
authors.forename: Manuel
authors.initials: M
authors.lastname: Tardáguila
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología/CSIC, Madrid.
authors.forename: Andrea
authors.initials: A
authors.lastname: Díaz-Alderete
authors.affiliation: Servicio de Reumatología, Instituto de Investigación Hospital 12 de Octubre, Madrid.
authors.forename: Alicia
authors.initials: A
authors.lastname: Usategui
authors.affiliation: Servicio de Reumatología, Instituto de Investigación Hospital 12 de Octubre, Madrid.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Miranda-Ramos
authors.affiliation: Bioinformatics Unit, Centro Nacional de Biotecnología/CSIC, Madrid.
authors.forename: Dannys Jorge
authors.initials: DJ
authors.lastname: Martínez-Herrera
authors.affiliation: Centro de Investigación Príncipe Felipe, Valencia.
authors.forename: Lorena
authors.initials: L
authors.lastname: de la Fuente
authors.affiliation: Centro de Biología Molecular Severo Ochoa/CSIC-UAM, Madrid.
authors.forename: María J
authors.initials: MJ
authors.lastname: García-León
authors.affiliation: Flow Cytometry Unit, Centro Nacional de Biotecnología/CSIC, Madrid.
authors.forename: María C
authors.initials: MC
authors.lastname: Moreno
authors.affiliation: Flow Cytometry Unit, Centro Nacional de Biotecnología/CSIC, Madrid.
authors.forename: Sara
authors.initials: S
authors.lastname: Escudero
authors.affiliation: Unitat d'Artritis, Hospital Clínic de Barcelona and Institut d'Investigacions Biomèdiques August Pí i Sunyer (IDIBAPS), Barcelona.
authors.forename: Juan D
authors.initials: JD
authors.lastname: Cañete
authors.affiliation: Centro de Biología Molecular Severo Ochoa/CSIC-UAM, Madrid.
authors.forename: María L
authors.initials: ML
authors.lastname: Toribio
authors.affiliation: Institut de Medicina Predictiva i Personalitzada del Càncer, Badalona, Barcelona, Spain.
authors.forename: Ildefonso
authors.initials: I
authors.lastname: Cases
authors.affiliation: Bioinformatics Unit, Centro Nacional de Biotecnología/CSIC, Madrid.
authors.forename: Alberto
authors.initials: A
authors.lastname: Pascual-Montano
authors.affiliation: Servicio de Reumatología, Instituto de Investigación Hospital 12 de Octubre, Madrid.
authors.forename: José Luis
authors.initials: JL
authors.lastname: Pablos
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología/CSIC, Madrid.
authors.forename: Santos
authors.initials: S
authors.lastname: Mañes
mesh_terms: D000368:Aged; D001172:Arthritis, Rheumatoid / Q000235:genetics* / Q000473:pathology; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator / Q000235:genetics / Q000378:metabolism*; D018920:Coculture Techniques; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D008264:Macrophages / Q000378:metabolism*; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D010003:Osteoarthritis / Q000235:genetics / Q000473:pathology; D051880:Receptors, Notch / Q000235:genetics*; D015398:Signal Transduction
chemical_list: D051784:Aryl Hydrocarbon Receptor Nuclear Translocator; D016207:Cytokines; D035683:MicroRNAs; D051880:Receptors, Notch; C497396:ARNT protein, human; C525067:MIR223, human
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