RNA as the catalyst for screening drugs against trypanosomatids

RNA作为筛选锥虫药物的催化剂

• 批准号: 7168987
• 负责人: REZA Salavati SALAVATI
• 金额: $ 13.4万
• 依托单位: MCGILL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-18 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7168987
• 关键词: NIH Roadmap Initiative tag; RNA; Trypanosoma; catalyst; chemical registry /resource; drug discovery /isolation; fluorescence resonance energy transfer; high throughput technology; mitochondria; nucleic acids; oxidative phosphorylation; posttranscriptional RNA processing; protozoal infection; uridine

DESCRIPTION (provided by applicant): The three related trypanosomatid pathogens, Trypanosoma brucei spec., Trypanosoma cruzi, and Leishmania spec., are the causative agents of African sleeping sickness, Chagas1 disease, and Leishmaniasis, respectively. These important pathogens affect large populations globally and cause numerous deaths. The drugs for these diseases are unsuitable since they are very toxic, not very effective, and resistance to these drugs has developed. Mitochondrial gene expression in trypanosomatids requires a particular form of RNA editing, unique to these organisms, by which precursor mRNA sequences are changed, often extensively, by the insertion and the deletion of uridine nucleotides. The edited mRNAs are translated into components of the oxidative phosphorylation system. Our finding that mitochondrial RNA editing is essential in the disease-causing stage of T. brucei has suggested a new promising target for anti- trypanosomatid drugs. Editing is catalyzed by a multiprotein complex (editosome) that has not yet been fully defined or characterized. This project proposes to generate chemical compounds to characterize the editosome proteins and determine their potential as targets for drugs that will be effective against the three related trypanosomatid pathogens. We are planning to use our expertise in the RNA biochemistry and adapt our ribozyme based assay for high throughput screening of compounds against the editosome. Specific Aim 1 outlines the plan to develop a high throughput screen using fluorescence resonance energy transfer (FRET) to report on the essential functions of the editosome. Specific Aim 2 will describe the secondary assays to validate the specificity of inhibition observed in the first aim. These studies will potentially identify compounds that may selectively perturb editosome assembly and/or function and characterize the various interactions and reaction stages of the RNA editing. They will also contribute to development of drugs against trypanosomatids.

性状（由申请方提供）：三种相关锥虫病原体，布氏锥虫，克氏锥虫和利什曼原虫，分别是非洲昏睡病、恰加斯病和利什曼病的病原体。这些重要的病原体影响全球大量人口，并导致许多人死亡。治疗这些疾病的药物是不合适的，因为它们毒性很大，不是很有效，并且对这些药物产生了耐药性。锥虫中的线粒体基因表达需要这些生物体特有的特定形式的RNA编辑，通过这种RNA编辑，前体mRNA序列通常通过尿苷核苷酸的插入和缺失而广泛改变。编辑的mRNA被翻译成氧化磷酸化系统的组分。我们发现线粒体RNA编辑在T.布氏锥虫病为抗锥虫病药物提供了一个新的靶点。编辑是由一种多蛋白复合物（编辑体）催化的，这种复合物尚未完全定义或表征。该项目提出生成化合物来表征编辑体蛋白，并确定其作为有效对抗三种相关锥虫病原体的药物靶点的潜力。我们正计划利用我们在RNA生物化学方面的专业知识，并调整我们的基于核酶的检测方法，以高通量筛选针对编辑体的化合物。具体目标1概述了使用荧光共振能量转移（FRET）开发高通量筛选以报告编辑体的基本功能的计划。具体目标2将描述验证第一个目标中观察到的抑制特异性的次要试验。这些研究将潜在地鉴定可能选择性地干扰编辑体组装和/或功能的化合物，并表征RNA编辑的各种相互作用和反应阶段。它们还将有助于开发抗锥虫药物。