IRAK-M Associates with Susceptibility to Adult-Onset Asthma and Promotes Chronic Airway Inflammation This information is current as Yi Liu, Mingqiang Zhang, Lili Lou, Lun Li, Youming of September 28, 2021. Zhang, Wei Chen, Weixun Zhou, Yan Bai and Jinming Gao J Immunol published online 7 January 2019 http://www.jimmunol.org/content/early/2019/01/04/jimmun ol.1800712 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2019/01/04/jimmunol.180071 Material 2.DCSupplemental http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! 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Published January 7, 2019, doi:10.4049/jimmunol.1800712 The Journal of Immunology IRAK-M Associates with Susceptibility to Adult-Onset Asthma and Promotes Chronic Airway Inflammation Yi Liu,*,†,1 Mingqiang Zhang,*,1 Lili Lou,* Lun Li,* Youming Zhang,‡ Wei Chen,x Weixun Zhou,{ Yan Bai,‖ and Jinming Gao* IL-1R–associated kinase (IRAK)-M regulates lung immunity during asthmatic airway inflammation. However, the regulatory effect of IRAK-M differs when airway inflammation persists. A positive association between IRAK-M polymorphisms with childhood asthma has been reported. In this study, we investigated the role of IRAK-M in the susceptibility to adult-onset asthma and in chronic airway inflammation using an animal model. Through genetic analysis of IRAK-M polymorphisms in a cohort of adult-onset asthma patients of Chinese Han ethnicity, we identified two IRAK-M single nucleotide polymorphisms, rs1624395 and rs1370128, genetically associated with adult-onset asthma. Functionally, the top-associated rs1624395, with an enhanced affinity to the transcription factor c-Jun, was associated with a higher expression of IRAK-M mRNA in blood monocytes. In contrast to the Downloaded from protective effect of IRAK-M in acute asthmatic inflammation, we found a provoking impact of IRAK-M on chronic asthmatic inflammation. Following chronic OVA stimulation, IRAK-M knockout (KO) mice presented with significantly less inflammatory cells, a lower Th2 cytokine level, a higher IFN-g concentration, and increased percentage of Th1 cells in the lung tissue than wild type mice. Moreover, lung dendritic cells (DC) from OVA-treated IRAK-M KO mice expressed a higher percentage of costim- ulatory molecules PD-L1 and PD-L2. Mechanistically, in vitro TLR ligation led to a greater IFN-g production by IRAK-M KO DCs than wild type DCs. These findings demonstrated a distinctive role of IRAK-M in maintaining chronic Th2 airway inflam- http://www.jimmunol.org/ mation via inhibiting the DC-mediated Th1 activation and indicated a complex role for IRAK-M in the initiation and progression of experimental allergic asthma. The Journal of Immunology, 2019, 202: 000–000. sthma is a heterogeneous disease characterized by of IL-1R–associated kinase (IRAK) gene on human chromosome chronic airway inflammation and airway remodeling 12q14.2 is one of the most consistently replicated regions linked A (1, 2), affecting patients of all ages (3, 4). Genome-wide to asthma or asthma-related traits in diverse populations (9, 10). A association studies and positional cloning have identified mul- positive association between IRAK-M SNPs and early-onset tiple single nucleotide polymorphisms (SNPs) of several asthma- asthma was reported in an Italian population (11). However, ge- by guest on September 28, 2021 associated genes expressing in airway epithelium, illustrating the netic studies on IRAK-M SNPs in asthma or atopy have critical role of the respiratory epithelial barrier in the pathogenesis yielded inconsistent results in children and adult patients in of asthma (5, 6). Epithelial cells, in conjunction with type 2 innate different populations (12–14). In addition to asthma, several lymphoid cells, dendritic cells (DCs), macrophages, and other cell IRAK-M SNPs have been implicated in other acute inflammatory types, constitute an innate immune network in the airway. Acti- diseases, such as acute lung injury (15). vation of this immune network regulates the Th2/Th17–dominant IRAK-M, primarily expressed by DCs, macrophages, and airway adaptive immune response (7). A predisposition to asthma was epithelium, is an intracellular negative regulator of TLRs that shown to be related to the impaired host innate immunity (8). regulates the innate immune homeostasis (11, 16). Evolving evi- Moreover, genome-wide association studies reveal that the locus dence has shown IRAK-M also participates in asthmatic immune *Department of Respiratory Diseases, Peking Union Medical College Hospital, Chinese tions; Y.B. reviewed the experimental data and edited the manuscript for important Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China; intellectual contents; Y.Z. designed the experiments, reviewed the experimental data, †Department of Respiratory Medicine, Civil Aviation General Hospital, Beijing 100123, and edited the manuscript for important intellectual contents; and J.G. designed and China; ‡Genomics Medicine Section, National Heart and Lung Institute, Imperial College supervised this study, interpreted the experimental findings, and drafted the manu- London, London SW3 6LY, United Kingdom; xDepartment of Cardiology, Peking Union script. All authors were involved in experimental design, data interpretation, and Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union manuscript preparation. All authors have read and approved the final version of this Medical College, Beijing 100730, China; {Department of Pathology, Peking Union Med- manuscript. ical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical ‖ Address correspondence and reprint requests to Prof. Jinming Gao, Department College, Beijing 100730, China; and Division of Pulmonary and Critical Care Medicine, of Respiratory Diseases, Peking Union Medical College Hospital, Number 1 Department of Internal Medicine, Brigham and Women’s Hospital, Harvard Medical Shuaifuyuan, Dongcheng District, Beijing 100730, China. E-mail address: School, Boston, MA 02115 [email protected] 1Y.L. and M.Z. equally contributed to this work. The online version of this article contains supplemental material. ORCIDs: 0000-0001-6226-1773 (L. Li); 0000-0001-5403-4044 (Y.Z.); 0000-0002- Abbreviations used in this article: a-SMA, a-smooth muscle actin; BAL, 7459-7343 (W.Z.); 0000-0002-8393-3062 (Y.B.). bronchoalveolar lavage; BMDC, bone marrow–derived DC; BMDM, bone marrow– Received for publication May 21, 2018. Accepted for publication November 30, derived macrophage; DC, dendritic cell; FEV1, forced expiratory volume in the first 2018. second; IRAK, IL-1R–associated kinase; KO, knockout; Mch, methacholine; MT, Masson trichrome; PAS, periodic acid–Schiff; PD-L1, programmed death–ligand 1; This work was supported by grants from National Natural Sciences Foundation of PD-L2, programmed death–ligand 2; qRT-PCR, quantitative real-time–PCR; SNP, single China (81170040, 81470229, and 30470767, all to J.G.) and National Institutes of nucleotide polymorphism; Treg, regulatory T; TSLP, thymic stromal lymphopoietin; WT, Health Grant K08HL135443 (to Y.B.). The sponsor had no role in study design, data wild type. collection and interpretation, and manuscript draft. Y.L., M.Z., and L. Lou performed the experiments; L. Li handled experimental Copyright Ó 2019 by The American Association of Immunologists, Inc. 0022-1767/19/$37.50 animals; W.C. facilitated the experiments; W.Z. performed the pathological evalua- www.jimmunol.org/cgi/doi/10.4049/jimmunol.1800712 2 IRAK-M AND CHRONIC ASTHMA responses. For instance, the expression of IRAK-M was upregu- Quantitative RT-PCR analysis for mRNA expression of lated in peripheral blood leukocytes in asthmatic children (8), IRAK-M in circulating monocytes in asthmatics and high mRNA expression of IRAK-M in sputum was a marker Total RNA was extracted from PBMCs in asthmatics using TRIzol reagents for frequent exacerbations of asthma (17). A unique feature of (Invitrogen) and then reverse transcribed into cDNA using M-MLV Reverse IRAK-M–mediated regulation is that it varies in a disease stage– Transcription System (Promega). Quantitative real-time PCR (qRT-PCR) dependent manner (18). Accumulating evidence from us and was employed to analyze mRNA expression of IRAK-M by a Bio-Rad others indicates that IRAK-M may exert contradictory roles, in- CFX96 Touch Real-Time PCR system using PowerUp SYBR Green Master Mix according to the manufacturer’s instructions (Applied Bio- hibitory or activating, in inflammatory and immune responses, systems). The housekeeping gene GAPDH was used as internal control to depending on the duration of stimulation and the kind of stimuli. normalize IRAK-M mRNA expression. All reactions