Cell Based Assays for Compounds That Regulate Tau Exon 10 Splicing

基于细胞的 Tau 外显子 10 剪接调节化合物测定

• 批准号: 7424262
• 负责人: JIANHUA ZHOU
• 金额: $ 16.25万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7424262
• 关键词: Alternative Splicing; Alzheimer' s Disease; Biological Assay; Brain; Cells; Chromosomes, Human, Pair 17; Data; Distal; Drug Compounding; Elements; Exons; Frontotemporal Dementia; Genes; Goals; Indium; Introns; Link; Messenger RNA; Microtubule-Associated Proteins; Microtubules; Monitor; Mutation; Neurodegenerative Disorders; Parkinsonian Disorders; Patients; Pharmaceutical Preparations; Production; Progressive Supranuclear Palsy; Protein Binding; Protein Isoforms; RNA Splicing; Reporter Genes; Screening procedure; System; Tauopathies; Testing; United States Food and Drug Administration; base; high throughput screening; prevent; small molecule; tau Proteins; tau aggregation

DESCRIPTION (provided by applicant): Tau aggregation is one of the most recognized features in more than 20 neurodegenerative disorders, classified as tauopathies that include Alzheimer's disease, frontotemporal dementia and Parkinsonism linked to chromosome 17 (FTDP-17) and progressive supranuclear palsy (PSP). More than 29 mutations in the tau gene have been identified from FTDP-17 patients. A group of these mutations alter splicing of exon 10, resulting in increase of exon 10 inclusion into the tau mRNA. In several other tauopathies such as PSP and Alzheimer's disease, abnormal splicing with inclusion of exon 10 into more tau mRNA has also been observed. These results indicate that one of the mechanisms that tau accumulates and aggregates in brains of tauopathies is abnormal splicing of the exon 10 towards to a more production of tau with the exon. Therefore, modulation of exon 10 splicing in the tau gene could be potentially targeted for preventing tauopathies. In order to establish a robust cell based system and to identify compounds/drugs that prevent exon 10 inclusion, we studied the mechanisms of tau splicing. We have demonstrated that a minimal distance between exons 10 and 11 is required for correct splicing of exon 10, and our additional preliminary data show that there are distal splicing cis-elements in intron 9. The goal of this proposal is to modify our existing mini reporter genes by including additional cis-elements and to establish a faithful cell based assay for identification of drugs that modulate exon 10 splicing. The assay will be optimized for high throughput screening and pilot screening will be carried out with 1040 FDA approved drugs and compounds.

描述（由申请人提供）：Tau聚集是20多种神经退行性疾病中最公认的特征之一，被归类为Tau病，包括阿尔茨海默病、额颞叶痴呆和17号染色体相关帕金森病（FTDP-17）和进行性核上性麻痹（PSP）。在FTDP-17患者中发现了超过29个tau基因突变。一组这些突变改变了外显子10的剪接，导致外显子10包含在tau mRNA中的增加。在其他一些tau病变中，如PSP和阿尔茨海默病，也观察到将外显子10包含到更多tau mRNA中的异常剪接。这些结果表明，tau在tau病患者大脑中积累和聚集的机制之一是外显子10的异常剪接，从而使tau与外显子产生更多。因此，调节tau基因的外显子10剪接可能是预防tau病的潜在目标。为了建立一个强大的基于细胞的系统，并识别阻止外显子10包合的化合物/药物，我们研究了tau剪接的机制。我们已经证明，外显子10和11之间需要最小的距离才能正确剪接，并且我们的额外初步数据表明，在内含子9中存在远端剪接顺式元件。本提案的目标是通过包括额外的顺式元件来修改我们现有的迷你报告基因，并建立一个忠实的基于细胞的检测方法