Development of a fluorescence-based technology to screen for RAGE-ligand binding inhibitors

The receptor for advanced glycation end-products (RAGE) and its ligands are important players in pathological conditions such as diabetes, neurodegenerative diseases, and cancers. RAGE is a cell surface molecule of the immunoglobulin superfamily. Alternatively spliced variants lacking either only the intracellular domain or both the intracellular and the transmembrane domains are also expressed in some tissues. RAGE functions in development and inflammation. It interacts with multiple ligands including members of the S100 protein family, high mobility group box-1 protein (HMGB1), and advanced glycation end products. It has been shown that the activation of RAGE triggers a positive feedback loop that stimulates the expression of both the receptor and some of its ligands, thus amplifying their deleterious effects (1-3). Therefore, the targeting of the specific interactions of RAGE with its ligands is a potential method to control these diseases. Toward this aim the project proposes an in vitro assay to screen for new molecules that block the interaction of the receptor with its pathological ligands.

The project aims to deliver purified unlabeled and fluorescently-labeled soluble RAGE and several of its ligands, a protocol for competitive binding assays based on the obtained products, and furthermore, an automated and validated technology which provides a rapid and easy way for screening of libraries of compounds.

References:

1. Yan SF, Ramasamy R, Schmidt AM. Receptor for AGE (RAGE) and its ligands- cast into leading roles in diabetes and the inflammatory response. J Mol Med (Berl). 2009 Mar;87(3):235-47.
2. A. Taguchi, D. C. Blood, G. del Toro, A. Canet, D. C. Lee, W. Qu, N. Tanji, Y. Lu, E. Lalla, C. Fu, M. A. Hofmann, T. Kislinger, M. Ingram, A. Lu, H. Tanaka, O. Hori, S. Ogawa, D. M. Stern, and A. M. Schmidt, "Blockade of RAGE-amphoterin signalling suppresses tumour growth and metastases," Nature, vol. 405, no. 6784, pp. 354-360, May 2000.
3. T. Arumugam, V. Ramachandran, and C. D. Logsdon, "Effect of cromolyn on S100P interactions with RAGE and pancreatic cancer growth and invasion in mouse models," J. Natl. Cancer Inst., vol. 98, no. 24, pp. 1806-1818, Dec. 2006.