High-throughput drug screen targeted to the 5′ untranslated region of Alzheimer amyloid precursor protein mRNA
High-throughput drug screen targeted to the 5′ untranslated region of Alzheimer amyloid precursor protein mRNA
复制标题
DOI：
10.1177/1087057106287271
复制
发表时间：
2006-08-01
影响因子：
--
通讯作者：
Rogers, Jack T.
中科院分区：
文献类型：
--
作者：
Bandyopadhyay, Sanghamitra;Ni, Jake;Rogers, Jack T.
作者采用了一种新方法来确定对阿尔茨海默病 (AD) 有效的治疗方法。 AD 淀粉样前体蛋白 (APP) mRNA 的 5' 非翻译区 (5'UTR) 是中枢神经系统中 APP 全蛋白和淀粉样β (Aβ) 肽水平的重要调节因子。作者生成了稳定的神经母细胞瘤 SH-SY5Y 转染子，该转染子在 146 核苷酸 APP mRNA 5'UTR 的翻译控制下表达荧光素酶。和由病毒内部核糖体进入位点驱动的绿色荧光蛋白（GFP）。他们使用高通量筛选 (HTS) 筛选了从神经变性药物发现实验室 (LDDN) 库中获得的 10,000 种化合物对 APP mRNA 5'UTR 控制的荧光素酶报告基因翻译的影响。该筛选产生了几种 APP mRNA 5'UTR 驱动的荧光素酶的无毒特异性抑制剂，对稳定 SH-SY5Y 转染子中的 GFP 表达没有影响。此外，这些化合物要么不抑制，要么抑制程度低得多，由朊病毒蛋白（PrP）mRNA 5'UTR调节的荧光素酶报告基因的表达，用作替代mRNA结构，在稳定转染的SH-SY5Y细胞系中反筛选APP mRNA 5'UTR。从这种稳健、特异性和高度定量的 HTS 中获得的命中结果将被表征，以鉴定可能被开发成有用的未来治疗剂的药物，以限制 AD 的 APP 翻译和 A beta 产生。
The authors employed a novel approach to identify therapeutics effective in Alzheimer disease (AD). The 5' untranslated region (5'UTR) of the mRNA of AD amyloid precursor protein (APP) is a significant regulator of the levels of the APP holo-protein and amyloid beta (A beta) peptide in the central nervous system. The authors generated stable neuroblastoma SH-SY5Y transfectants that express luciferase under the translational control of the 146-nucleotide APP mRNA 5'UTR. and green fluorescent protein (GFP) driven by a viral internal ribosomal entry site. Using a high-throughput screen (HTS), they screened for the effect of I 10,000 compounds obtained from the library of the Laboratory for Drug Discovery on Neurodegeneration (LDDN) on the APP mRNA 5'UTR-controlled translation of the luciferase reporter. This screening yielded several nontoxic specific inhibitors of APP mRNA 5'UTR-driven luciferase that had no effect on the GFP expression in the stable SH-SY5Y transfectants. Moreover, these compounds either did not inhibit or inhibited to a much lower extent the expression of the luciferase reporter regulated by a prion protein (PrP) mRNA 5'UTR, used as an alternative mRNA structure to counterscreen APP mRNA 5'UTR in stably transfected SH-SY5Y cell lines. The hits obtained from this robust, specific, and highly quantitative HTS will be characterized to identify agents that may be developed into useful future therapeutic agents to limit APP translation and A beta production for AD.