(AFS) RNA Editing Ligase 1 and F1-ATPase as novel drug targets for veterinary diseases caused by trypanosomatid parasites

(AFS) RNA 编辑连接酶 1 和 F1-ATPase 作为锥虫寄生虫引起的兽医疾病的新药物靶点

• 批准号: 2672460
• 金额: --
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2672460
• 关键词: AFS; RNA; Editing; Ligase; F1

The goal of this project is to exploit the unique and essential functions of mitochondrial RNA editing and F1-ATPase in trypanosomatid parasites as targets for the identification of much-needed new drugs. Our validation of Trypanosoma brucei RNA editing ligase 1 (TbREL1) and F1-ATPase as drug targets (Schnaufer et al., 2001; 2005), and the availability of a high-resolution crystal structures (Deng et al., 2004; Montgomery et al., 2018) and high-throughput screening assays make these enzymes promising and ripe targets for drug discovery efforts.The protist parasites Trypanosoma congolense, T. vivax and T. brucei brucei are responsible for devastating economic loss by causing disease in livestock (http://www.fao.org/ag/againfo/programmes/en/paat/home.html). All trypanosomatids are characterised by unusual mitochondrial biology. For example, they require a unique form of RNA editing for mitochondrial gene expression; a key enzyme in this process is REL1 (Schnaufer et al., 2001). Also, T. brucei uses the mitochondrial F1-ATPase not to generate ATP, but as an essential proton pump (Schnaufer et al., 2005). Orthologues of these enzymes in T. congolense, T. vivax and T. brucei are virtually identical, which offers exciting potential for the development of inhibitors with broad anti-trypanosomatid activity. We recently carried out a number of high-throughput screens for small molecule inhibitors of REL1 and F1-ATPase, and we identified and confirmed numerous structurally diverse inhibitors with low-micromolar IC50 values.This PhD project will involve participating in further hit identification and in hit-to-lead development, specifically testing inhibitors in vitro (activity assays; biophysical characterisation using ITC and surface plasmon resonance; toxicology studies) and in vivo (against trypanosomes and control cells) and structural analysis of target-inhibitor complexes by crystallography, cryo-EM and/or computational predictions. Key collaborators on this project are Dr Atlanta Cook (2nd supervisor; Wellcome Centre for Cell Biology), an expert in the structural analysis of proteins, Prof Manfred Auer, an expert in chemical biology (Centre for Systems Biology), and Prof Asier Unciti-Broceta (Edinburgh Cancer Research UK Centre), an expert in medicinal chemistry. The combined expertise of the collaborating labs provides an ideal environment for the interdisciplinary training of PhD students.

该项目的目标是利用锥虫寄生虫中线粒体 RNA 编辑和 F1-ATP 酶的独特且重要的功能作为鉴定急需的新药物的靶标。我们对布氏锥虫 RNA 编辑连接酶 1 (TbREL1) 和 F1-ATPase 作为药物靶点的验证（Schnaufer 等人，2001 年；2005 年），以及高分辨率晶体结构的可用性（Deng 等人，2004 年；Montgomery 等人） al., 2018）和高通量筛选分析使这些酶成为药物发现工作有希望且成熟的目标。原生寄生虫刚果锥虫、间日锥虫和布氏锥虫通过引起牲畜疾病而造成毁灭性的经济损失 (http://www.fao.org/ag/againfo/programmes/en/paat/home.html)。所有锥虫都具有不寻常的线粒体生物学特征。例如，它们需要一种独特形式的 RNA 编辑来表达线粒体基因；该过程中的一个关键酶是 REL1（Schnaufer 等，2001）。此外，T. brucei 使用线粒体 F1-ATP 酶不是为了产生 ATP，而是作为重要的质子泵（Schnaufer 等，2005）。刚果锥虫、间日锥虫和布氏锥虫中这些酶的直系同源物实际上是相同的，这为开发具有广泛抗锥虫活性的抑制剂提供了令人兴奋的潜力。我们最近对REL1和F1-ATP酶的小分子抑制剂进行了一些高通量筛选，我们鉴定并确认了许多具有低微摩尔IC50值的结构多样的抑制剂。这个博士项目将涉及参与进一步的命中鉴定和研究hit-to-lead 开发，特别是在体外（活性测定；使用 ITC 和表面等离振子共振进行生物物理表征；毒理学研究）和体内（针对锥虫和对照细胞）和结构分析测试抑制剂通过晶体学、冷冻电镜和/或计算预测来分析靶标-抑制剂复合物。该项目的主要合作者包括蛋白质结构分析专家Atlanta Cook 博士（第二任导师；Wellcome 细胞生物学中心）、化学生物学专家Manfred Auer 教授（系统生物学中心）和Asier Unciti 教授。 Broceta（英国爱丁堡癌症研究中心），药物化学专家。合作实验室的综合专业知识为博士生的跨学科培训提供了理想的环境。