HTS Molecules Targeting APP 5'untranslated Region(RMI)

阿尔茨海默病淀粉样前体蛋白(APP)5‘端非翻译区是关键 翻译监管元素，用于设置任何给定单元格中的应用程序生产量。IL-1增强血管生成的作用 铁调节蛋白(IRPS)与APP 5‘非编码区RNA二级结构的相互作用 响应元件(IRE)。这一规定表明APP 5‘UTR是一个很好的药物靶点。 我们的实验室在筛选FDA的文库时，在药物发现领域获得了宝贵的经验- 预先批准的药物，并确定了17个线索，限制了APP 5‘UTR驱动的翻译使用瞬时 转基因神经母细胞瘤细胞。在基于二次蛋白质印迹的分析中，帕罗西汀(SSRI)和 二硫代丙醇(螯合剂)选择性降低APP全蛋白表达。作为药物选择性的证据 在APP 5‘UTR屏幕上实现的这两个点击没有改变APLP-1和APLP-2在 SH-SY5Y细胞(Payton等人，2003年)。一项初步研究表明，帕罗西汀减少了淀粉样蛋白的负荷 转基因AD小鼠模型(TgCRNDS小鼠)。 通过R03 RFA机制收集的数据将帮助我们开发FDA试点筛查，以 建立一种基于高通量转基因的重要RNA靶标筛选方法。使用自动化的 通过我们与药物实验室的合作，现在可以获得384个基于良好基础的筛分的设备 发现神经退行性变(LDDN)。我们的战略将是优化高吞吐量的筛分 LDDN药物库中的110,000种化合物用于识别选择性抑制翻译驱动的新药物 由APP 5‘UTR增强器。这些导联很可能提供下游治疗抗淀粉样蛋白限制。 行动。在基于RNA凝胶移动和APP表达的实验中使用我们的药物HITS将探索 APP翻译路径。IRP-1和IRP-2转录后调控铁稳态 铁蛋白翻译与转铁蛋白受体mRNA稳定性。HTS针对146个NT应用程序5‘UTR的点击量将 提供新的工具来准确评估IRP-1和IRP-2与5‘UTR相互作用的重要性 编码APP(金属蛋白)的mRNA，相对于其他关键铁蛋白的mRNAs。

The 5'untranslated region of the mRNAforthe Alzheimer's Amyloid Precursor Protein (APP 5'UTR) is a key translational regulatory element that sets the amount of APP production in any given cell. IL-1 enhanced the interaction between Iron-regulatory Proteins (IRPs) and APP 5'UTR RNA secondary structure via an Iron- responsive Element (IRE). This regulation suggested the APP 5'UTR to be an excellent drug target. Our laboratory gained valuable experience in the field of drug discovery when we screened a library of FDA- pre-approved drugs and identified 17 leads that limited APP 5'UTR driven translation using transiently transfected neuroblastoma cells. In secondary western blot-based assays paroxetine (SSRI) and dimercaptopropanol (chelator) selectivley reduced APP holoprotein expression. As proof of drug selectivity achieved during the APP 5'UTR screen these two hits did not change APLP-1 and APLP-2 expression in SH-SY5Y cells (Payton et al.,2003). A pilot study indicated that Paroxetine reduced the amyloid burden in a transgenic mouse model for AD (TgCRNDS mice). The data to be gathered through this R03 RFA mechanism will assist us to develop our FDA pilot screen to set up a high throughput transfection based screen of an important RNA target. Use of automated equipment for 384 well based screens is now available through our collaboration with the Laboratory of Drug Discovery for Neurodegenreration (LDDN). Our strategy will be to optimize a high throughput screen of the 110,000 compounds in the LDDN drug library to identify new agents that selectively inhibit translation driven by the APP 5'UTR enhancer. These leads may well provide downstream therapeutic anti-amyloid limiting action. Use of our drug hits in RNA gel-shift and APP expression based experiments will probe the pathway of APP translation. IRP-1 and IRP-2 post-transcriptionally control iron homeostasis by modulating ferritin translation and transferrin receptor mRNA stability. HTS hits directed to the 146 nt APP 5'UTR will provide new tools to assess precisely the importance of the interaction of IRP-1 and IRP-2 with 5'UTR of the mRNA encodiing APP (metalloprotein), relative to the mRNAs of other key iron proteins.