(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酶的独特和基本功能，作为鉴定急需的新药的靶点。我们验证了布氏锥虫RNA编辑连接酶1（TbN 1）和F1-ATP酶作为药物靶标（Schnaufer等人，2001; 2005）和高分辨率晶体结构的可用性（Deng等人，2004;蒙哥马利等人，2018）和高通量筛选测定使这些酶成为药物发现工作的有希望和成熟的靶标。间日疟原虫和T.布鲁氏菌病是造成牲畜疾病的主要原因，造成了毁灭性的经济损失（http：//www.fao.org/ag/againfo/programmes/en/paat/home.html）。所有锥虫的特点是不寻常的线粒体生物学。例如，它们需要一种独特形式的RNA编辑来进行线粒体基因表达;该过程中的一种关键酶是BMP 1（Schnaufer et al. 2001年）。所以，T。布氏杆菌使用线粒体F1-ATP酶不产生ATP，而是作为必需的质子泵（Schnaufer等，2005年）。这些酶在T. congolense，T.间日疟原虫和T.布氏锥虫几乎是相同的，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。我们最近进行了一些高通量筛选的小分子抑制剂的F1和F1-ATP酶，我们确定并确认了许多结构不同的抑制剂与低微摩尔IC 50值。这个博士项目将涉及参与进一步的命中识别和命中铅的发展，特别是在体外测试抑制剂（活性测定;使用ITC和表面等离子体共振的生物物理表征;毒理学研究）和体内研究（针对锥虫和对照细胞）和通过晶体学、冷冻-EM和/或计算预测的靶标-抑制剂复合物的结构分析。该项目的主要合作者是蛋白质结构分析专家Atlanta Cook博士（第二主管; Wellcome细胞生物学中心），化学生物学专家Manfred奥尔教授（系统生物学中心）和药物化学专家Asier Unciti-Broceta教授（爱丁堡癌症研究英国中心）。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。