Fluorescent RNA foci assay for FXTAS drug discovery

用于 FXTAS 药物发现的荧光 RNA 焦点测定

• 批准号: 8647780
• 负责人: Karen Wu
• 金额: $ 20.79万
• 依托单位: LUCERNA, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8647780
• 关键词: 5' Untranslated Regions; Adult; Affect; Age; Ataxia; Biological Assay; CGG repeat; Caring; Cell Death; Cell Line; Cell Nucleus; Cell model; Cell physiology; Cells; Cerebellar Gait; Characteristics; Cognitive; Communities; Data; Development; Disease; Disease Progression; Dyes; Elements; Ensure; Environment; Exhibits; FMR1 Gene; FXTAS; Fluorescence; Fluorescence Microscopy; Foundations; Future; General Population; Image; Impaired cognition; Individual; Kinetics; Label; Life; Measures; Neurodegenerative Disorders; Noise; Nuclear; Nuclear Inclusion; Nuclear RNA; Parkinson Disease; Parkinsonian Disorders; Pathology; Performance; Pharmaceutical Preparations; Phase; Protein Splicing; Proteins; RNA; RNA Sequences; RNA Splicing; RNA-Binding Proteins; Recording of previous events; Reporter; Reproducibility; Signal Transduction; Small Business Innovation Research Grant; Spinach - dietary; Staging; Structure; Symptoms; System; Techniques; Technology; Therapeutic; Toxic effect; Transcript; Tremor; Trinucleotide Repeat Expansion; Trinucleotide Repeats; Woman; base; disability; drug discovery; fluorophore; gain of function; high throughput screening; inhibitor/antagonist; men; novel; prevent; public health relevance; research study; screening; small molecule; small molecule libraries; stable cell line; validation studies

PROJECT SUMMARY ! Fragile X-associated tremor/ataxia syndrome (FXTAS) is a progressive neurodegenerative disorder that that predominately affects adults over the age of 50. Individuals afflicted with FXTAS develop tremor, ataxia, Parkinson-like symptoms, and slowly lose their cognitive abilities to the point that they are completely dependent on others for care. Currently, there is no cure. Therefore, there is a strong need to discover first-in-class drug treatments capable of preventing and/or reversing FXTAS disease progression. FXTAS is a trinucleotide repeat disorder caused by premutations in the fragile X mental retardation 1 (FMR1) gene. RNA gain-of-function toxicity from the expanded trinucleotide tracts in FMR1 is thought to be the primary cause of FXTAS pathology. In the nucleus, expanded trinucleotide tracts form RNA aggregates that sequester RNA-binding proteins and splicing factors from normal cellular functions and trigger cell degeneration. Therefore, the discovery of small molecules that disrupt the nuclear formation of toxic RNA aggregates would be highly desirable as potential therapeutics for FXTAS. In order to discover ribonuclear foci inhibitors, high-throughput assays capable of providing a convenient readout for RNA foci formation are needed. Current techniques suffer from major limitations that make them difficult to use and produce results of dubious validity. In this proposal, we will develop a RNA-based trinucleotide repeat RNA reporter system that will enable us to detect the formation of CGG150 RNA foci in a cellular environment and facilitate the screening of small molecules libraries for drugs that inhibit toxic CGG150 RNA aggregation. !

项目摘要 呢 脆弱的X相关震颤/共济失调综合征（FXTA）是一种渐进式 主要影响50岁以上成年人的神经退行性疾病。 患有FXTA的人会出现震颤，共济失调，帕金森氏症症状和 慢慢失去认知能力，以至于他们完全取决于 其他人要照顾。目前，无法治愈。因此，很有必要发现 能够预防和/或逆转FXTAS病的第一类药物治疗 进展。 FXTA是由预言引起的三核苷酸重复障碍 脆弱的X智力低下1（FMR1）基因。 RNA功能获得的毒性 FMR1中的扩展的三核苷酸区域被认为是FXTA的主要原因 病理。在细胞核中，膨胀的三核苷酸区域形成RNA聚集体 来自正常细胞功能的隔离RNA结合蛋白和剪接因子 触发细胞变性。因此，发现破坏的小分子 有毒RNA聚集体的核形成将是高度可取的 FXTA的治疗剂。为了发现核糖核焦点抑制剂，高通量 需要为RNA焦点形成提供方便的读数的测定。 当前的技术受到主要局限性，使它们难以使用和 产生可疑有效性的结果。在此提案中，我们将开发基于RNA的 三核苷酸重复RNA报告基因系统，该系统将使我们能够检测到形成 CGG150 RNA焦点在细胞环境中，并促进筛选小 分子文库用于抑制有毒CGG150 RNA聚集的药物。 呢