针对β-分泌酶1适配子的筛选及其联合AChE的适配子治疗AD的实验研究

项目名称： 针对β-分泌酶1适配子的筛选及其联合AChE的适配子治疗AD的实验研究

项目编号： No.81471388

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 张兴梅

作者单位： 南方医科大学

项目金额： 70万元

中文摘要： 对抗Aβ形成及其毒性是治疗AD的一种根本策略。BACE1是Aβ生成过程中的关键酶，抑制BACE1活性可从源头上阻断Aβ的产生。SELEX技术可筛选出与靶蛋白的高特异和高亲和力结合的抑制剂，是一种新型的药物筛选方法。本课题采用SELEX技术筛选抑制BACE1胞外区蛋白活性的适配子（aptamer）。目前最常用药物就是AChE抑制剂，多靶标联合用药可提高AD治疗效果。我们已筛选出具有抑制作用的AChE适配子。所以我们尝试将BACE1适配子单独或与联合AChE适配子分别作用于稳定表达APPsw、APPwt的AD细胞模型，观察其对细胞的作用和Aβ产量的影响；将适配子与纳米颗粒结合，观察其对AD动物模型的行为学改变及脑内Aβ和老年斑生成等变化。同时还探讨BACE1适配子在AD早期诊断中的作用。本课题运用SELEX技术来进行探索诊断和治疗AD的新方法和新思路，为AD诊疗相关基础研究提供新的技术平台

中文关键词： 阿尔茨海默病；适配子；β-分泌酶1；乙酰胆碱酯酶；诊断和治疗

英文摘要： Suppression of the formation of amyloid-beta (Aβ) and its toxicity is the fundamental strategy in the treatment of Alzheimer's disease (AD). β-site amyloid precursor protein cleaving enzyme 1 (BACE1) plays a crucial role in the process of Aβ generation. Consequently, one way to prevent Aβ generation from the source maight be to inhibit the activity of BACE1. The systematic evolution of ligands by exponential enrichment (SELEX) technology, which can select inhibitors that bind to their target proteins with high specificity and good affinity, is considered a new method of drug screening. In this study, we aim to adopt SELEX technology to select aptamers able to inhibit the extracelluar region of BACE1 activity specifically. Currently acetylcholine esterase (AChE) inhibitors are the most common clinical drugs for the treatment of AD. Based on the consensus that multi-target combination therapy can improve therapeutic effect because of the multi-factor pathogenic mechanism of AD and the fact that anti-AChE aptamers have been selected in our previous report, we attempt to evaluate the effect of anti-BACE1 aptamer alone or combined with anti-AChE aptamer on cells stably transfected with APPsw or APPwt genes and on Aβ production in these cellular models of AD. Next we try to incorparate these aptamers into nanoparticles to observe its influence on behavior changes and pathological changes of Aβ and senile plaque generation in AD animal models. Simultaneously we will investigate the role of anti-BACE1 aptamer on early diagnosis of AD. In our study SELEX technology is applied to explore new approach and ideas for AD diagnosis and treatment, which provides a new technology platform for correlational research of AD.

英文关键词： Alzheimer's Disease;aptamer;BACE1;AChE;diagnosis and therapy