Carboxylated Glycans Mediate Colitis through Activation of NF- B

Glycobiology Program, The Burnham Institute, La Jolla, CA 92037, USA.
The Journal of Immunology (Impact Factor: 4.92). 11/2005; 175(8):5412-22. DOI: 10.4049/jimmunol.175.8.5412
Source: PubMed


The role of carbohydrate modifications of glycoproteins in leukocyte trafficking is well established, but less is known concerning how glycans influence pathogenesis of inflammation. We previously identified a carboxylate modification of N-linked glycans that is recognized by S100A8, S100A9, and S100A12. The glycans are expressed on macrophages and dendritic cells of normal colonic lamina propria, and in inflammatory infiltrates in colon tissues from Crohn's disease patients. We assessed the contribution of these glycans to the development of colitis induced by CD4(+)CD45RB(high) T cell transfer to Rag1(-/-) mice. Administration of an anti-carboxylate glycan Ab markedly reduced clinical and histological disease in preventive and early therapeutic protocols. Ab treatment reduced accumulation of CD4(+) T cells in colon. This was accompanied by reduction in inflammatory cells, reduced expression of proinflammatory cytokines and of S100A8, S100A9, and receptor for advanced glycation end products. In vitro, the Ab inhibited expression of LPS-elicited cytokines and induced apoptosis of activated macrophages. It specifically blocked activation of NF-kappaB p65 in lamina propria cells of colitic mice and in activated macrophages. These results indicate that carboxylate-glycan-dependent pathways contribute to the early onset of colitis.

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Available from: Simon H Murch, Sep 04, 2014
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    • "We also found that inhibiting carboxylated glycandependent interactions of DAMP molecules using mAbGB3.1, an anti-glycan antibody blocked onset of T cell mediated colitis [Srikrishna et al., 2005b], colitis-dependent colon cancer [Turovskaya et al., 2008] and recruitment of myeloid-derived suppressor cells (MDSC) to secondary lymphoid organs in tumor-bearing mice [Sinha et al., 2008]. "
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    ABSTRACT: The receptor for advanced glycation end products (RAGE) is a signaling receptor protein of the immunoglobulin superfamily implicated in multiple pathologies. It binds a diverse repertoire of ligands, but the structural basis for the interaction of different ligands is not well understood. We earlier showed that carboxylated glycans on the V-domain of RAGE promote the binding of HMGB1 and S100A8/A9. Here we study the role of these glycans on the binding and intracellular signaling mediated by another RAGE ligand, S100A12. S100A12 binds carboxylated glycans, and a subpopulation of RAGE enriched for carboxylated glycans shows more than 10-fold higher binding potential for S100A12 than total RAGE. When expressed in mammalian cells, RAGE is modified by complex glycans predominantly at the first glycosylation site (N25IT) that retains S100A12 binding. Glycosylation of RAGE and maximum binding sites for S100A12 on RAGE are also cell type dependent. Carboxylated glycan-enriched population of RAGE forms higher order multimeric complexes with S100A12, and this ability to multimerize is reduced upon deglycosylation or by using non-glycosylated sRAGE expressed in E. coli. mAbGB3.1, an antibody against carboxylated glycans, blocks S100A12-mediated NF-kappaB signaling in HeLa cells expressing full-length RAGE. These results demonstrate that carboxylated N-glycans on RAGE enhance binding potential and promote receptor clustering and subsequent signaling events following oligomeric S100A12 binding.
    Journal of Cellular Biochemistry 06/2010; 110(3):645-59. DOI:10.1002/jcb.22575 · 3.26 Impact Factor
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    • "Glycosylation of the extracellular part of RAGE significantly increases receptor affinity for HMGB1 and S100 proteins. In addition, carboxylated glycans were found on a subset of RAGE on colon cancer cells [51,52]. "
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    ABSTRACT: The receptor for advanced glycation end products (RAGE) is a single transmembrane receptor of the immunoglobulin superfamily that is mainly expressed on immune cells, neurons, activated endothelial and vascular smooth muscle cells, bone forming cells, and a variety of cancer cells. RAGE is a multifunctional receptor that binds a broad repertoire of ligands and mediates responses to cell damage and stress conditions. It activates programs responsible for acute and chronic inflammation, and is implicated in a number of pathological diseases, including diabetic complications, stroke, atheriosclerosis, arthritis, and neurodegenerative disorders. The availability of Rage knockout mice has not only advanced our knowledge on signalling pathways within these pathophysiological conditions, but also on the functional importance of the receptor in processes of cancer. Here, we will summarize molecular mechanisms through which RAGE signalling contributes to the establishment of a pro-tumourigenic microenvironment. Moreover, we will review recent findings that provide genetic evidence for an important role of RAGE in bridging inflammation and cancer.
    Cell Communication and Signaling 06/2009; 7(1):12. DOI:10.1186/1478-811X-7-12 · 3.38 Impact Factor
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    • "It was shown that different extracellular molecules such as heparin sulphate proteoglycans were able to bind S100A12 and other S100 proteins [38]. RAGE receptor – the primary S100A12 target – is not uniformly glycosylated in vivo and only a small proportion of RAGE molecules are able to form a productive complex with its ligands [39,40], (Srikrishna & Freeze, unpublished results). Therefore our further experiments were focused on in vitro studies to demonstrate that zinc/calcium-dependent oligomerization of S100A12 may be an additional and very important factor contributing to its target interactions; this was probed on the example of the best known interaction partner, RAGE. "
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    ABSTRACT: Human S100A12 is a member of the S100 family of EF-hand calcium-modulated proteins that are associated with many diseases including cancer, chronic inflammation and neurological disorders. S100A12 is an important factor in host/parasite defenses and in the inflammatory response. Like several other S100 proteins, it binds zinc and copper in addition to calcium. Mechanisms of zinc regulation have been proposed for a number of S100 proteins e.g. S100B, S100A2, S100A7, S100A8/9. The interaction of S100 proteins with their targets is strongly dependent on cellular microenvironment. The aim of the study was to explore the factors that influence S100A12 oligomerization and target interaction. A comprehensive series of biochemical and biophysical experiments indicated that changes in the concentration of calcium and zinc led to changes in the oligomeric state of S100A12. Surface plasmon resonance confirmed that the presence of both calcium and zinc is essential for the interaction of S100A12 with one of its extracellular targets, RAGE--the Receptor for Advanced Glycation End products. By using a single-molecule approach we have shown that the presence of zinc in tissue culture medium favors both the oligomerization of exogenous S100A12 protein and its interaction with targets on the cell surface. We have shown that oligomerization and target recognition by S100A12 is regulated by both zinc and calcium. Our present work highlighted the potential role of calcium-binding S100 proteins in zinc metabolism and, in particular, the role of S100A12 in the cross talk between zinc and calcium in cell signaling.
    BMC Biochemistry 05/2009; 10(1):11. DOI:10.1186/1471-2091-10-11 · 1.44 Impact Factor
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