Soluble Receptor for Advanced Glycation End Products for Therapeutic Application

通过晚期糖基化终末产物受体(RAGE)的细胞信号转导导致促炎反应。RAGE介导的炎症与糖尿病、动脉粥样硬化和阿尔茨海默病等炎症性疾病有关。RAGE的剪接或蛋白降解形式被称为可溶性RAGE(SRAGE)，其功能是作为一种天然的诱饵来对抗RAGE信号。动物模型已经证明，给药可阻止动脉粥样硬化的形成，并稳定血管壁上现有的斑块。此外，SRAGE还可以防止血管损伤引起的新生内膜的形成，从而抑制再狭窄。 我们已经建立了稳定表达sRAGE的中国仓鼠卵巢(CHO)细胞系，以及产生同源sRAGE的伴随亲和纯化策略。系统地研究了SRAGE在再狭窄动物模型中的应用，并对数据进行了分析。我们的结果表明，我们实验室生产的SRAGE的效价是以前报道的1000倍。除了阻断再狭窄，我们还在脑梗塞动物模型上测试了Sage阻断，并获得了有希望的初步结果。我们还进行了研究，以探索观察到的高效性的分子基础，并发现SRAGE中的N-糖链结构与其生物活性有关。我们目前正在与约翰霍普金斯大学合作，确定N-糖链的轮廓和结构，旨在进一步增强SRAGE的生物活性。 为了进一步将SRAGE开发为一种有效的治疗产品，我们还与能源部的Ames实验室合作，启动了使用基于介孔二氧化硅纳米颗粒的药物输送系统的研究，以实现更有效的体内释放和控制释放。我们希望开发携带sRAGE的纳米粒传递-释放系统，与其他抗炎和抗血栓药物联合使用，以有效对抗血管疾病。该试剂目前正在开发中，不能用于公共或商业用途。 为了克服表达和检测sRAGE的技术障碍，我们还开发了一套便于亚克隆、细胞表面表达的表达模块。以及哺乳动物膜蛋白的表位标记。这项发明已经获得了美国临时专利(第61/142,531号)，NIH目前正在为这项发明做广告。研发现状：临床前体外试验。

Cellular signaling via receptor for advanced glycation end products (RAGE) results in pro-inflammatory responses. RAGE-mediated inflammation has been implicated in inflammatory diseases including diabetes, atherosclerosis, and Alzheimers disease. The spliced or proteolytically cleaved form of RAGE is referred as soluble RAGE (sRAGE), which functions as a natural decoy counter-effecting RAGE signaling. It has been demonstrated in animal models that administration of sRAGE blocks atherogenesis, and stabilizes existing plaques on the vessel wall. In addition, sRAGE also prevents the formation of neointima prompted by vascular injuries and hence inhibits restenosis. We have developed Chinese Hamster Ovary (CHO) cell lines that stably express sRAGE, and the accompanied affinity purification strategies that produce homogenous sRAGE. Systemic studies of sRAGE application in restenosis animal models have been completed, and data have been analyzed. Our results showed that sRAGE produced in our laboratory exhibits 1000 x higher potency than that of previously reported. In addition to blocking restenosis, we also tested sRAGE blockage on infarct animal models and obtained promising preliminary results. We also performed studies to explore the molecular basis of the observed high potency of sRAGE and found that N-glycan structure in sRAGE contributes to its bioactivity. We are currently collaborating with Johns Hopkins University to determine the N-glycan profile and structure, aiming at further potentiating sRAGE bioactivity. To further develop sRAGE as an effective therapeutic product, we, in collaboration with Department of Energy's Ames Laboratory, also initiated studies using mesoporous silica nanoparticle-based drug delivery system for a more effective in vivo delivery, and controlled release. We hope to develop nanoparticle delivery-release system carrying sRAGE in combination with other anti-inflammatory and anti-thrombosis drug to effectively combat vascular diseases. The reagent is currently under development, and is not available for public or commercial usage. To overcome technical hurdles for expression and detection of sRAGE, we also developed a set of expression modules that facilitate subcloning, cell-surface expression. and epitope tagging of mammalian membrane proteins. U.S. Provisional Patent (No. 61/142,531) has been awarded to this invention, and NIH is currently advertising the invention. R&D Status: Pre-clinical in vitro.