The Receptor for Advanced Glycation End Products Is a Central Mediator of Asthma Pathogenesis

Abstract

The receptor for advanced glycation end products (RAGE) is a multiligand receptor that has been shown to contribute to the pathogenesis of diabetes, atherosclerosis, and neurodegeneration. However, its role in asthma and allergic airway disease is largely unknown. These studies use a house dust mite (HDM) mouse model of asthma/allergic airway disease. Respiratory mechanics were assessed and compared between wild-type and RAGE knockout mice. Bronchovascular architecture was assessed with quantitative scoring, and expression of RAGE, immunoglobulins, and relevant cytokines was assessed by standard protein detection methods and/or quantitative RT-PCR. The absence of RAGE abolishes most assessed measures of pathology, including airway hypersensitivity (resistance, tissue damping, and elastance), eosinophilic inflammation, and airway remodeling. IL-4 secretion, isotype class switching, and antigen recognition are intact in the absence of RAGE. In contrast, normal increases in IL-5, IL-13, eotaxin, and eotaxin-2 production are abrogated in the RAGE knockouts. IL-17 indicates complex regulation, with elevated baseline expression in RAGE knockouts, but no induction in response to allergen. Treatment of WT mice with an inhibitor of RAGE markedly reduces inflammation in the HDM model, suggesting that RAGE inhibition may serve as a promising therapeutic strategy. Finally, the results in the HDM model are recapitulated in an ovalbumin model of asthma, suggesting that RAGE plays a role in asthma irrespective of the identity of the allergens involved. The receptor for advanced glycation end products (RAGE) is a multiligand receptor that has been shown to contribute to the pathogenesis of diabetes, atherosclerosis, and neurodegeneration. However, its role in asthma and allergic airway disease is largely unknown. These studies use a house dust mite (HDM) mouse model of asthma/allergic airway disease. Respiratory mechanics were assessed and compared between wild-type and RAGE knockout mice. Bronchovascular architecture was assessed with quantitative scoring, and expression of RAGE, immunoglobulins, and relevant cytokines was assessed by standard protein detection methods and/or quantitative RT-PCR. The absence of RAGE abolishes most assessed measures of pathology, including airway hypersensitivity (resistance, tissue damping, and elastance), eosinophilic inflammation, and airway remodeling. IL-4 secretion, isotype class switching, and antigen recognition are intact in the absence of RAGE. In contrast, normal increases in IL-5, IL-13, eotaxin, and eotaxin-2 production are abrogated in the RAGE knockouts. IL-17 indicates complex regulation, with elevated baseline expression in RAGE knockouts, but no induction in response to allergen. Treatment of WT mice with an inhibitor of RAGE markedly reduces inflammation in the HDM model, suggesting that RAGE inhibition may serve as a promising therapeutic strategy. Finally, the results in the HDM model are recapitulated in an ovalbumin model of asthma, suggesting that RAGE plays a role in asthma irrespective of the identity of the allergens involved. The receptor for advanced glycation end products (RAGE) is a multiligand receptor first identified as a potential mediator in diabetes.1Neeper M. Schmidt A.M. Brett J. Yan S.D. Wang F. Pan Y.C. Elliston K. Stern D. Shaw A. Cloning and expression of a cell surface receptor for advanced glycosylation end products of proteins.J Biol Chem. 1992; 267: 14998-15004Abstract Full Text PDF PubMed Google Scholar RAGE has many additional ligands, including S100 proteins, HMGB1, amyloid β, and heparin.2Hanford L.E. Enghild J.J. Valnickova Z. Petersen S.V. Schaefer L.M. Schaefer T.M. Reinhart T.A. Oury T.D. Purification and characterization of mouse soluble receptor for advanced glycation end products (sRAGE).J Biol Chem. 2004; 279: 50019-50024Abstract Full Text Full Text PDF PubMed Scopus (188) Google Scholar, 3Sparvero L.J. Asafu-Adjei D. Kang R. Tang D. Amin N. Im J. Rutledge R. Lin B. Amoscato A.A. Zeh H.J. Lotze M.T. RAGE (Receptor for Advanced Glycation Endproducts), RAGE ligands, and their role in cancer and inflammation.J Transl Med. 2009; 7: 17Crossref PubMed Scopus (461) Google Scholar The current paradigm maintains that membrane RAGE (mRAGE) signaling is proinflammatory, whereas soluble RAGE (sRAGE), a secreted form of RAGE, is generally anti-inflammatory because it scavenges proinflammatory ligands.4Ramasamy R. Yan S.F. Schmidt A.M. RAGE: therapeutic target and biomarker of the inflammatory response–the evidence mounts.J Leukoc Biol. 2009; 86: 505-512Crossref PubMed Scopus (236) Google Scholar RAGE transcript and protein are predominantly expressed in the lung,5Brett J. Schmidt A.M. Yan S.D. Zou Y.S. Weidman E. Pinsky D. Nowygrod R. Neeper M. Przysiecki C. Shaw A. et al.Survey of the distribution of a newly characterized receptor for advanced glycation end products in tissues.Am J Pathol. 1993; 143: 1699-1712PubMed Google Scholar and specifically by pulmonary type I alveolar epithelial cells,6Englert J.M. Hanford L.E. Kaminski N. Tobolewski J.M. Tan R.J. Fattman C.L. Ramsgaard L. Richards T.J. Loutaev I. Nawroth P.P. Kasper M. Bierhaus A. Oury T.D. A role for the receptor for advanced glycation end products in idiopathic pulmonary fibrosis.Am J Pathol. 2008; 172: 583-591Abstract Full Text Full Text PDF PubMed Scopus (166) Google Scholar suggesting that RAGE has an important role in lung pathophysiology. Recent studies suggest that RAGE contributes to pulmonary disease; RAGE knockout mice are protected against hyperoxia-induced lung injury7Reynolds P.R. Schmitt R.E. Kasteler S.D. Sturrock A. Sanders K. Bierhaus A. Nawroth P.P. Paine III, R. Hoidal J.R. Receptors for advanced glycation end-products targeting protect against hyperoxia-induced lung injury in mice.Am J Respir Cell Mol Biol. 2010; 42: 545-551Crossref PubMed Scopus (69) Google Scholar and exhibit attenuated responses to bacterial pneumonia.8Ramsgaard L. Englert J.M. Manni M.L. Milutinovic P.S. Gefter J. Tobolewski J. Crum L. Coudriet G.M. Piganelli J. Zamora R. Vodovotz Y. Enghild J.J. Oury T.D. Lack of the receptor for advanced glycation end-products attenuates E. coli pneumonia in mice.PLoS One. 2011; 6: e20132Crossref PubMed Scopus (44) Google Scholar RAGE also appears to have an important role in pulmonary fibrosis, but this depends on the type of injury model used to generate the fibrotic response.6Englert J.M. Hanford L.E. Kaminski N. Tobolewski J.M. Tan R.J. Fattman C.L. Ramsgaard L. Richards T.J. Loutaev I. Nawroth P.P. Kasper M. Bierhaus A. Oury T.D. A role for the receptor for advanced glycation end products in idiopathic pulmonary fibrosis.Am J Pathol. 2008; 172: 583-591Abstract Full Text Full Text PDF PubMed Scopus (166) Google Scholar, 9Englert J.M. Kliment C.R. Ramsgaard L. Milutinovic P.S. Crum L. Tobolewski J.M. Oury T.D. Paradoxical function for the receptor for advanced glycation end products in mouse models of pulmonary fibrosis.Int J Clin Exp Pathol. 2011; 4: 241-254PubMed Google Scholar, 10He M. Kubo H. Ishizawa K. Hegab A.E. Yamamoto Y. Yamamoto H. Yamaya M. The role of the receptor for advanced glycation end-products in lung fibrosis.Am J Physiol Lung Cell Mol Physiol. 2007; 293: L1427-L1436Crossref PubMed Scopus (135) Google Scholar, 11Ramsgaard L. Englert J.M. Tobolewski J. Tomai L. Fattman C.L. Leme A.S. Kaynar A.M. Shapiro S.D. Enghild J.J. Oury T.D. The role of the receptor for advanced glycation end-products in a murine model of silicosis.PLoS One. 2010; 5: e9604Crossref PubMed Scopus (30) Google Scholar RAGE expression is altered in these models of disease, for example, the appearance of sRAGE in the bronchoalveolar lavage fluid (BALF) in a mouse model of pneumonia and loss of RAGE expression in models of pulmonary fibrosis. Asthma/allergic airway disease (AAD) is a main inflammatory condition of modern industrial societies, seen with increasing frequency throughout the developing world. Bronchodilators and corticosteroids remain the mainstays of therapy, but they are ineffective or inadequate for some groups of patients. Novel therapies that exploit auxiliary mechanisms of disease and that incur fewer side effects than chronic corticosteroid treatment are greatly needed. To date, there appears to have been no studies of RAGE in animal models of asthma. A few studies in humans have suggested that there is an increase in the levels of RAGE ligands HMGB112Watanabe T. Asai K. Fujimoto H. Tanaka H. Kanazawa H. Hirata K. Increased levels of HMGB-1 and endogenous secretory RAGE in induced sputum from asthmatic patients.Respir Med. 2011; 105: 519-525Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar and S100A8/A913Halayko A.J. Ghavami S. S100A8/A9: a mediator of severe asthma pathogenesis and morbidity?.Can J Physiol Pharmacol. 2009; 87: 743-755Crossref PubMed Scopus (67) Google Scholar in samples from patients with asthma compared with controls, suggesting that RAGE may contribute to asthma/AAD pathogenesis. Although one recent study suggested that sRAGE is increased concomitantly with HMGB1 in patients with asthma,12Watanabe T. Asai K. Fujimoto H. Tanaka H. Kanazawa H. Hirata K. Increased levels of HMGB-1 and endogenous secretory RAGE in induced sputum from asthmatic patients.Respir Med. 2011; 105: 519-525Abstract Full Text Full Text PDF PubMed Scopus (84) Google Scholar another suggested a decrease in sRAGE and no change in HMGB1 in patients with neutrophilic asthma.14Sukkar M.B. Wood L.G. Tooze M. Simpson J.L. McDonald V.M. Gibson P.G. Wark P.A. Soluble RAGE is deficient in neutrophilic asthma and chronic obstructive pulmonary disease.Eur Respir J. 2012; 39: 721-729Crossref PubMed Scopus (108) Google Scholar Apart from the potential inconsistency between the latter two results, those studies have not provided mechanistic insight as to the role of mRAGE versus sRAGE in asthma, nor have they elucidated how cytokines and chemokines to allergic disease are in the or absence of RAGE. The study used a house dust mite (HDM) antigen model of asthma/AAD to the role of RAGE in of this model is that and are by HDM in the absence of HDM antigen has been identified as a to asthma pathogenesis in humans and is a of asthmatic The role of in of PubMed Scopus Google Scholar, dust mite allergens in asthma and Mol Med. 2010; Full Text Full Text PDF PubMed Scopus Google Scholar To the of the results and to were also in an ovalbumin model of to that effects of RAGE are not to the model mice to these models airway to and cell with and elevated of the class most with allergic disease. WT mice were from RAGE knockout mice were provided by A. Bierhaus of and from these mice a was J.M. Hanford L.E. Kaminski N. Tobolewski J.M. Tan R.J. Fattman C.L. Ramsgaard L. Richards T.J. Loutaev I. Nawroth P.P. Kasper M. Bierhaus A. Oury T.D. A role for the receptor for advanced glycation end products in idiopathic pulmonary fibrosis.Am J Pathol. 2008; 172: 583-591Abstract Full Text Full Text PDF PubMed Scopus (166) Google Scholar, 11Ramsgaard L. Englert J.M. Tobolewski J. Tomai L. Fattman C.L. Leme A.S. Kaynar A.M. Shapiro S.D. Enghild J.J. Oury T.D. The role of the receptor for advanced glycation end-products in a murine model of silicosis.PLoS One. 2010; 5: e9604Crossref PubMed Scopus (30) Google Scholar, R. A. B. H.J. Nawroth B. of a mouse to as by a mouse PubMed Scopus Google Scholar These mice are with the RAGE mice were and to WT mice for In mice were in the animal of the of and were by the and HDM was from was from sRAGE was from mouse lung and was with a as L.E. Enghild J.J. Valnickova Z. Petersen S.V. Schaefer L.M. Schaefer T.M. Reinhart T.A. Oury T.D. Purification and characterization of mouse soluble receptor for advanced glycation end products (sRAGE).J Biol Chem. 2004; 279: 50019-50024Abstract Full Text Full Text PDF PubMed Scopus (188) Google Scholar, J.M. Ramsgaard L. Valnickova Z. Enghild J.J. Oury T.D. and of soluble RAGE from lung 2008; PubMed Scopus Google Scholar was by and of the sRAGE to a RAGE HMGB1, was assessed as N. C. Kasper M. M. L. of cell and a function of RAGE, the of alveolar epithelial type I PubMed Scopus Google Scholar to was intact in the was induced in mice with the use of one of In the first WT or RAGE mice were for with of HDM in of mice were with were the In the WT mice were for with one of the in of of of mouse sRAGE of HDM of HDM with of or of HDM with of The used ovalbumin as the was by treatment with of ovalbumin with of in of on and mice were with with of ovalbumin in of in on for a of for were the function was with a as L.M. M. airway in allergic airway J Respir Med. 2007; PubMed Scopus Google Scholar mice were with and and were to the were with a and of of and were and were by to a model of the was a the response was by for a of response were for of that lung pulmonary function the mice were and were with the use of was in the the and cell were and were for cell were with and and were on the The of and were by fluid and were for was from lung with the use of an the was with murine in a was with and with for RAGE and The was on an and of by The change in RAGE, IL-5, or compared with was with the Lung were by and mouse in with by protein protein in lung was with the of lung of or of were by and as of protein and of Scopus Google Scholar in were in by studies that the antigen of from and of HDM was by and to and in were in from and WT or RAGE mice by were with detection was with for and and by and with an against on lung were a on with and with was in with for with or were were with in whereas and were in in To RAGE, in against mouse sRAGE was by to from the animal in the RAGE was and RAGE lung were used to were with a of were with and by an were with with 2004; Scholar were as by In a of was were with with or of and by were with were In a of was eotaxin-2 from from IL-17 and were or in for IL-17 used lung was by with HDM in were with with or lung was and with was by with to and were and were as was with an the samples were in HDM was first with the use of was with a were with of with HDM was to was by to and The were and were as lung from mice were by a been to the identity of the The of in lung was and inflammation was in was as or The results are expressed as the of that inflammatory in as an by the of the inflammation by the of in that was with and quantitative results were expressed as was with of A was to To RAGE expression was altered in mice and with HDM compared with controls, and on lung were Although there was a decrease in RAGE transcript expression in the of mice with HDM compared with this was not change was in the expression of RAGE protein or in the of sRAGE versus two of RAGE, of protein of the receptor for advanced glycation end-products expressed in murine and cell 2009; PubMed Scopus Google Scholar and mRAGE Finally, sRAGE not in or of or mice RAGE is expressed by type I alveolar epithelial it was that in RAGE expression in cell with as epithelial or inflammatory by lung expression studies of lung to this no change in or RAGE expression in response to HDM and and expression in type I alveolar epithelial of a mediator not altered in a disease the mediator is a receptor with to the absence of RAGE or RAGE mice on the were to the HDM WT mice for function in airway function in WT mice with HDM with an asthmatic These in to were in the to airway airway tissue and tissue The change in compared with is seen in studies in was with of studies in the K. A.S. A. of in the airway and its role in allergic airway J Physiol Lung Cell Mol Physiol. 2010; PubMed Scopus Google Scholar The with the was with a elevated to in RAGE mice compared with WT but no between versus HDM RAGE mice was there were no in airway or architecture in RAGE it is that in in this are to a of RAGE or by on airway function for the potential role of However, it is important to that RAGE mice with HDM and from those of WT and RAGE mice. of markedly elevated cell and in the allergic WT mice compared with of lung and in of allergic WT mice by whereas cell and elevated expression of in of the allergic WT mice compared with In to WT RAGE mice no inflammatory and no elevated expression or cell These results suggested that RAGE plays a role in the tissue in including airway eosinophilic and airway remodeling. To the response to HDM and was altered in RAGE was on and lung of WT and RAGE mice to the levels of and elevated levels of in and lung of WT and RAGE mice that been and with HDM were To there were between the HDM antigen of from and WT versus RAGE HDM by and to were with from and WT and RAGE mice no in antigen between the two the levels of and were elevated in the of WT and RAGE mice and with HDM compared with their IL-4 is a in cell type as as in isotype class to J.J. and its of the allergic Clin Scopus Google Scholar To how the absence of RAGE the production of this in response to HDM and was on The IL-4 response was to intact in RAGE as there was no between RAGE and WT mice in the expression of this in response to Although not a of the IL-17 has been suggested to a mediator relevant to asthma, to with a neutrophilic in asthma and 2011; PubMed Scopus Google Scholar, D. I. Tan Z. F. M. 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Full Text Full Text PDF PubMed Scopus Google Scholar To of the two mRAGE or was the allergic in the HDM mouse model, and to sRAGE has a therapeutic has been seen in many disease in WT mice and with HDM sRAGE from mouse was HDM In to a protein an of was with HDM in a of mice. as by cell and was markedly in mice that sRAGE HDM compared with mice that HDM or HDM lung were by a to treatment for the of the of inflammatory and a of the of the inflammation in The of inflammation and the inflammation were in mice sRAGE HDM compared with the two HDM the cell A of these studies was to the role of RAGE in murine models of asthma/AAD on the identity of the the ovalbumin model of asthma/AAD was used to this because the antigen and markedly from that of the HDM was with the use of ovalbumin to the as in the HDM function in the and in WT mice with ovalbumin compared with mice with to in the HDM model Although the change in the not the in the and cell were elevated in allergic WT and was and eosinophilic inflammation and were in allergic WT mice compared with RAGE mice have airway function and no or inflammatory or These suggested that role in murine asthma/AAD is of the involved. The results of this study suggest that RAGE is a mediator in the disease mechanisms that in airway and airway and pulmonary in an HDM mouse model of the in response of the WT versus RAGE mice to HDM it is that RAGE levels and in the pulmonary to between and the that it is not RAGE but a protein expression is to RAGE, that is for the effects seen in RAGE mice. Although this the that treatment with sRAGE for RAGE specifically and markedly reduces inflammation in response to HDM that RAGE than a in is for the the sRAGE treatment studies that of the RAGE signaling of or may have in the treatment of The suggest that and response mechanisms that are intact in the absence of RAGE, including antigen and production of and isotype class switching, this latter by Although are to the main of IL-5, and IL-13, cell have been to the production of one or of this IL-4 induction in the absence of the two cytokines of in the response the or the of cell that may as E. 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The and that of and airway as and IL-13, are abrogated in the absence of RAGE, suggesting that RAGE as a mediator in one or proinflammatory that in the altered and seen in this model of of its RAGE has for some been by many to a recognition To date, no has been in the ovalbumin model of asthma suggest that RAGE mice are to the tissue and seen in WT mice with hypersensitivity to pulmonary eosinophilic inflammation, and airway remodeling. the in the and the between the HDM and ovalbumin and the of the response against HDM in RAGE it is to that RAGE of antigen its ligands in asthma/AAD are to endogenous and not from the Finally, RAGE is predominantly expressed on alveolar type I epithelial some studies have suggested that it is expressed on and as of the cell for the seen in these studies those with the as as of the between RAGE and the cytokines and IL-13, of Bierhaus for with the RAGE mouse