MAFB Determines Human Macrophage Anti-Inflammatory Polarization: Relevance for the Pathogenic Mechanisms Operating in Multicentric Carpotarsal Osteolysis This information is current as of September 26, 2021. Víctor D. Cuevas, Laura Anta, Rafael Samaniego, Emmanuel Orta-Zavalza, Juan Vladimir de la Rosa, Geneviève Baujat, Ángeles Domínguez-Soto, Paloma Sánchez-Mateos, María M. Escribese, Antonio Castrillo, Valérie Cormier-Daire, Miguel A. Vega and Ángel L. Corbí Downloaded from J Immunol published online 16 January 2017 http://www.jimmunol.org/content/early/2017/01/15/jimmun ol.1601667 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2017/01/15/jimmunol.160166 Material 7.DCSupplemental Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 26, 2021 • No Triage! 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Published January 16, 2017, doi:10.4049/jimmunol.1601667 The Journal of Immunology MAFB Determines Human Macrophage Anti-Inflammatory Polarization: Relevance for the Pathogenic Mechanisms Operating in Multicentric Carpotarsal Osteolysis Vı´ctor D. Cuevas,* Laura Anta,† Rafael Samaniego,‡ Emmanuel Orta-Zavalza,* Juan Vladimir de la Rosa,x Genevie`ve Baujat,{,‖ A´ ngeles Domı´nguez-Soto,* Paloma Sa´nchez-Mateos,‡ Marı´a M. Escribese,# Antonio Castrillo,x Vale´rie Cormier-Daire,{,‖ Miguel A. Vega,* and A´ ngel L. Corbı´* Macrophage phenotypic and functional heterogeneity derives from tissue-specific transcriptional signatures shaped by the local micro- environment. Most studies addressing the molecular basis for macrophage heterogeneity have focused on murine cells, whereas the factors Downloaded from controlling the functional specialization of human macrophages are less known. M-CSF drives the generation of human monocyte-derived macrophages with a potent anti-inflammatory activity upon stimulation. We now report that knockdown of MAFB impairs the acquisition of the anti-inflammatory profile of human macrophages, identify the MAFB-dependent gene signature in human macrophages and il- lustrate the coexpression of MAFB and MAFB-target genes in CD163+ tissue-resident and tumor-associated macrophages. The con- tribution of MAFB to the homeostatic/anti-inflammatory macrophage profile is further supported by the skewed polarization of monocyte-derived macrophages from multicentric carpotarsal osteolysis (Online Mendelian Inheritance in Man #166300), a pathology http://www.jimmunol.org/ caused by mutations in the MAFB gene. Our results demonstrate that MAFB critically determines the acquisition of the anti- inflammatory transcriptional and functional profiles of human macrophages. The Journal of Immunology, 2017, 198: 000–000. acrophage heterogeneity derives from the existence of bition of tumor cell growth and production of LPS-induced cy- tissue-specific factors that control macrophage differ- tokines (7, 14). Specifically, GM-CSF primes macrophages M entiation and functional maturation. M-CSF and IL-34 (GM-MØ) to gain immunogenic activity and to produce inflam- control macrophage differentiation in most tissues (1, 2), whereas matory cytokines upon TLR stimulation, whereas M-CSF primes GM-CSF drives the generation of alveolar macrophages (3). Cir- macrophages (M-MØ) with tissue repair and proangiogenic culating monocytes are recruited to damaged tissues under in- functions, and with potent TLR-induced IL-10–producing ability by guest on September 26, 2021 flammatory conditions (4) and acquire specialized functions (7, 15). Accordingly, human GM-MØ and M-MØ are considered (macrophage polarization) under the influence of extracellular as proinflammatory and anti-inflammatory macrophages (6, 16). cues (5). The macrophage sensitivity to the surrounding milieu is MAFB is a transcription factor of the large MAF subfamily exemplified by the ability of GM-CSF and M-CSF to induce the (MAFA, cMAF, MAFB, NRL) that binds to a specific DNA acquisition of distinct effector functions (6, 7). Previous studies element (MARE); heterodimerizes with cMAF, JUN, and FOS have demonstrated that GM-CSF–primed macrophages (GM-MØ) (17–19); and associates and functionally inhibits MYB (19), and M-CSF–primed macrophages (M-MØ) exhibit distinct cellu- MITF, and NFATc1 (20). MAFB controls lens development (21), lar phenotypes (8–11), a distinct metabolic state (12, 13), and lymphangiogenesis (22), pancreatic a and b cell differentiation (23, display opposite effector functions like activin A–mediated inhi- 24), skin cell differentiation (25), chondrocyte matrix formation and *Laboratorio de Ce´lulas Mieloides, Centro de Investigaciones Biolo´gicas, Consejo Grant S2010/BMD-2350 (to A´ .L.C. and A.C.), and a Formacio´ndePersonalInves- Superior de Investigaciones Cientı´ficas, 28040 Madrid, Spain; †Servicio de Cirugı´a tigador predoctoral fellowship from MINECO through Grant BES-2012-053864 Ortope´dica y Traumatologı´a, Complejo Hospitalario de Santiago de Compostela, (to V.D.C.). 15706 Santiago de Compostela, Spain; ‡Laboratorio de Inmuno-Oncologı´a, Unidad V.D.C., L.A., M.M.E., A.C., M.A.V., and A´ .L.C. designed the research; V.D.C., L.A., de Microscopı´a Confocal, Instituto de Investigacio´n Sanitaria Gregorio Maran˜o´n, x R.S., E.O.-Z., J.V.d.l.R., G.B., A´ .D.-S., P.S.-M., and V.C.-D. performed experiments, 28007 Madrid, Spain; Instituto de Investigaciones Biomedicas Alberto Sols, Consejo { recruited patients, and analyzed data; V.D.C. and A´ .L.C. wrote the manuscript. Superior de Investigaciones Cientı´ficas, 28029 Madrid, Spain; Unidad de Biomedi- cina, Instituto de Investigaciones Biome´dicas–Universidad de Las Palmas de Gran The sequences presented in this article have been submitted to Gene Expression Canaria (ULPGC), Instituto Universitario de Investigaciones Biomedicas y Sanitarias Omnibus under accession number GSE84622. de la ULPGC, 35001 Las Palmas, Spain; ‖De´partement de Ge´ne´tique, INSERM Address correspondence and reprint requests to Dr. A´ ngel L. Corbı´ and Dr. Miguel A. U781, Universite´ Paris Descartes-Sorbonne Paris Cite´, Institut Imagine, Hoˆpital Vega, Centro de Investigaciones Biolo´gicas, Consejo Superior de Investigaciones Necker Enfants Malades, 75015 Paris, France; and #Institute for Applied Molecular Cientı´ficas, Calle Ramiro de Maeztu 9, 28040 Madrid, Spain. E-mail addresses: Medicine, School of Medicine, University CEU San Pablo, Madrid, Spain [email protected] (A´ .L.C.) and [email protected] (M.A.V.) ORCIDs: 0000-0002-2816-8070 (V.D.C.); 0000-0002-4196-5218 (L.A.); 0000-0002- The online version of this article contains supplemental material. 3081-7332 (R.S.); 0000-0003-1443-7548 (J.V.d.l.R.); 0000-0003-1980-5733 (A´ .L.C.). Abbreviations used in this article: GM-MØ, GM-CSF–primed macrophages; GSEA, Received for publication September 26, 2016. Accepted for publication December gene set enrichment analysis; MARE, MAF recognition element; MCTO, multicen- 16, 2016. tric carpotarsal osteolysis; M-MØ, M-CSF–primed macrophages; RANKL, receptor This work was supported by Ministerio de Economı´a y Competitividad (MINECO) activator for NF-kB ligand; siControl, small interfering RNA control; siMAFB, Grant SAF2014-52423-R and Instituto de Salud Carlos III Red de Investigacio´n MAFB-specific small interfering RNA; TRAP, tartrate-resistant acid phosphatase. en Enfermedades Reuma´ticas Grant RIER RD12/009 (to A´ .L.C. and M.A.V.), MINECO Grant SAF2014-56819-R (to A.C.), Comunidad Auto´noma de Madrid/ Copyright Ó 2017 by The American Association of Immunologists, Inc. 0022-1767/17/$30.00 Fonds Europe´en de De´veloppement E´ conomique et Re´gional RAPHYME Program www.jimmunol.org/cgi/doi/10.4049/jimmunol.1601667 2 MAFB-DIRECTED MACROPHAGE ANTI-INFLAMMATORY POLARIZATION development (26), and podocyte generation (27–30), and also reg- that were provided by M. Palomero (Oncology Department, Hospital Gen- ulates type I IFN production through recruitment of coactivators to eral Universitario Gregorio Maran˜o´n). Macrophage supernatants were IFN regulatory factor 3 (31). Within the murine myeloid lineage, assayed for the presence of cytokines using commercial ELISA kits for CCL2 (BD Biosciences) and IL-10 (BioLegend), according to the proto- MafB is preferentially expressed in most tissue-resident macro- cols supplied by the manufacturers. Mouse bone marrow–derived macro- phages, whose specific enhancers contain an overrepresentation of phages were generated using human M-CSF (10 ng/ml; ImmunoTools). All MARE sequences (32). MAFB restricts the ability of M-CSF to animal procedures were approved by the Comite´ E´ tico de Experimentacio´n instruct myeloid cell proliferation, promotes macrophage differen- Animal (Ethical Committee for Animal Experimentation) of the Consejo Superior de Investigaciones Cientı´ficas and conducted in accordance with tiation (33) through repression of self-renewal