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The N-glycoform of sRAGE is the key determinant for its therapeutic efficacy to attenuate injury-elicited arterial inflammation and neointimal growth

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Abstract

Signaling of the receptor for advanced glycation end products (RAGE) has been implicated in the development of injury-elicited vascular complications. Soluble RAGE (sRAGE) acts as a decoy of RAGE and has been used to treat pathological vascular conditions in animal models. However, previous studies used a high dose of sRAGE produced in insect Sf9 cells (sRAGESf9)and multiple injections to achieve the therapeutic outcome. Here, we explore whether modulation of sRAGE N-glycoform impacts its bioactivity and augments its therapeutic efficacy. We first profiled carbohydrate components of sRAGE produced in Chinese hamster Ovary cells (sRAGECHO) to show that a majority of its N-glycans belong to sialylated complex types that are not shared by sRAGESf9. In cell-based NF-κB activation and vascular smooth muscle cell (VSMC) migration assays, sRAGECHO exhibited a significantly higher bioactivity relative to sRAGESf9 to inhibit RAGE alarmin ligand-induced NF-κB activation and VSMC migration. We next studied whether this N-glycoform-associated bioactivity of sRAGECHO is translated to higher in vivo therapeutic efficacy in a rat carotid artery balloon injury model. Consistent with the observed higher bioactivity in cell assays, sRAGECHO significantly reduced injury-induced neointimal growth and the expression of inflammatory markers in injured vasculature. Specifically, a single dose of 3 ng/g of sRAGECHO reduced neointimal hyperplasia by over 70 %, whereas the same dose of sRAGESf9 showed no effect. The administered sRAGECHO is rapidly and specifically recruited to the injured arterial locus, suggesting that early intervention of arterial injury with sRAGECHO may offset an inflammatory circuit and reduce the ensuing tissue remodeling. Our findings showed that the N-glycoform of sRAGE is the key determinant underlying its bioactivity and thus is an important glycobioengineering target to develop a highly potent therapeutic sRAGE for future clinical applications.

Key message

  • The specific N-glycoform modification is the key underlying sRAGE bioactivity

  • Markedly reduced sRAGE dose to attenuate neointimal hyperplasia and inflammation

  • Provide a molecular target for glycobioengineering of sRAGE as a therapeutic protein

  • Blocking RAGE alarmin ligands during acute injury phase offsets neointimal growth

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Acknowledgments

We thank Robert Monticone for rat VSMCs and advice on cell migration assays. We also thank reviewers of this manuscript for their inputs that improve our work. The work was supported by the intramural research program of the NIH, National Institute on Aging (LL, MIT, RPX, and EGL), and the Korea Research Foundation grant KRF-2009-013-E00008 (SP). WW was supported in part by the Oak Ridge Institute for Science and Education’s Research Associates Program at NIH; JP was a recipient of the 2011 Johns Hopkins “Excellent in Medical Student Research Award”; and RAR and DB were supported by intramural research training awards from NIH.

Conflict of interest

NIH has filed a patent based on this work. The authors (SP, WW, R-P X, MIT, EGL, or LL) have not received any royalties from this patent.

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Authors and Affiliations

  1. Laboratory of Cardiovascular Science, National Institute on Aging, 251 Bayview Boulevard, Baltimore, MD, 21224, USA

    Hyun-Jin Tae, Ji Min Kim, Sungha Park, Wen Wei, Natalia Petrashevskaya, Ismayil Ahmet, John Pang, Stefanie Cruschwitz, Rebecca A. Riebe, Christopher H. Morrell, David Browe, Rui-ping Xiao, Mark I. Talan, Edward G. Lakatta & Li Lin

  2. Department of Biomedical Science and Research Institute for Bioscience and Biotechnology, Hallym University, Chunchon, South Korea

    Hyun-Jin Tae

  3. Division of Cardiology, Cardiovascular Center, Yonsei University College of Medicine, Seoul, South Korea

    Sungha Park

  4. Department of Biology, Johns Hopkins University, Baltimore, MD, USA

    Noboru Tomiya & Yuan Chuan Lee

  5. Institute of Molecular Medicine, Peking University, Beijing, People’s Republic of China

    Geng Li & Rui-ping Xiao

  6. Department of Physiology, University of Maryland School of Medicine, Baltimore, MD, USA

    Yinghua Zhang

  7. Department of Mathematics and Statistics, Loyola University, Baltimore, MD, USA

    Christopher H. Morrell

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  1. Hyun-Jin Tae
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Correspondence to Li Lin.

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Hyun-Jin Tae, Ji Min Kim, Sungha Park, and Noboru Tomiya contributed equally in this study.

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Tae, HJ., Kim, J.M., Park, S. et al. The N-glycoform of sRAGE is the key determinant for its therapeutic efficacy to attenuate injury-elicited arterial inflammation and neointimal growth. J Mol Med 91, 1369–1381 (2013). https://doi.org/10.1007/s00109-013-1091-4

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