Soluble Receptor for Advanced Glycation End Products for Therapeutic Application

用于治疗应用的高级糖基化终产物的可溶性受体

• 批准号: 8736641
• 负责人: Edward Lakatta
• 金额: $ 44.3万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8736641
• 关键词: Adenoviruses; Advanced Glycosylation End Products; Advertising; Affinity Chromatography; Algorithms; Alzheimer' s Disease; Amino Acid Sequence; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Atherosclerosis; Basic Science; Biochemical; Blood Vessels; Cardiovascular Diseases; Cell Line; Cells; Chinese Hamster Ovary Cell; Cleaved cell; Clinical Trials; Codon Nucleotides; Complementary DNA; Data; Development; Diabetes Mellitus; Disease; Dose; Drug Delivery Systems; Exhibits; Future; Genetic Engineering; Goals; Guanine + Cytosine Composition; Infarction; Inflammation; Inflammatory; Inflammatory Response; Injury; Insecta; Laboratories; Legal patent; Mediating; Molecular; Mus; Peptide Sequence Determination; Pharmaceutical Preparations; Phenotype; Polysaccharides; Post-Translational Protein Processing; Process; Production; Property; RNA Splice Sites; RNA Splicing; Recombinants; Signal Transduction; Staging; Structure; System; Testing; Therapeutic; United States National Institutes of Health; Vascular Diseases; Vascular Endothelial Cell; age related; atherogenesis; base; clinical application; combat; design; in vivo; monocyte; nanoparticle; neointima formation; particle; prevent; receptor; restenosis; scale up; tool; vector

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 to 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 accompanying affinity purification strategies that produce homogenous sRAGE. We also generated recombinant Adenovirus that expresses 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 500-1000 x higher efficacy than that of sRAGE produced in insect sf9 cells. 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. To further develop sRAGE as an effective therapeutic product, we used GeneOptimizer algorithm from Invitrogen to optimize T7-sRAGE----this tool removes sequence repeat, killer motifs, splice sites and RNA secondary structures in the cDNA sequence and optimize codon usage (for CHO cell) and GC content without changing protein sequence. We also tested and found that the new sRAGE cDNA has a higher level of expression than that of native cDNA. This step should enhance future sRAGE scale-up production. A patent (PCT/US2013/020103) based on sRAGE has been filed by NIH, and the OTT is currently advertising the invention. RAGE (KO) is at early stage (designing targeting vector).

通过受体的细胞信号传导晚期糖基化最终产物（RAGE）导致促炎反应。愤怒介导的炎症与包括糖尿病，动脉粥样硬化和阿尔茨海默氏病在内的炎症性疾病有关。愤怒的剪接或蛋白水解裂片形式称为可溶性愤怒（SRAGE），它是自然诱饵反应的愤怒信号传导。在动物模型中已经证明了SRAGE的给药可阻止动脉粥样硬化，并稳定了容器壁上现有的斑块。此外，Srage还防止了血管损伤引起的新内膜的形成，因此抑制再狭窄。 我们已经开发了稳定表达SRAGE的中国仓鼠卵巢（CHO）细胞系，以及产生同质Srage的随附的亲和力纯化策略。我们还产生了表达SRAGE的重组腺病毒。在再狭窄动物模型中应用SRAGE应用的系统性研究已经完成，并且已经分析了数据。我们的结果表明，在我们的实验室中产生的SRAGE比在昆虫SF9细胞中产生的SRAGE表现出500-1000 x的功效。除了阻塞再狭窄外，我们还测试了梗死动物模型上的SRAGE阻塞，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力的分子基础，并发现Srage中的N-聚糖结构有助于其生物活性。 To further develop sRAGE as an effective therapeutic product, we used GeneOptimizer algorithm from Invitrogen to optimize T7-sRAGE----this tool removes sequence repeat, killer motifs, splice sites and RNA secondary structures in the cDNA sequence and optimize codon usage (for CHO cell) and GC content without changing protein sequence.我们还测试并发现新的Srage cDNA的表达水平高于天然cDNA。此步骤应提高未来的SRAGE扩展生产。 NIH已提交了基于SRAGE的专利（PCT/US2013/020103），OTT目前正在宣传本发明。 RAGE（KO）处于早期（设计目标向量）。