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

该项目的目标是开发锥虫寄生虫线粒体RNA编辑和F1-ATPase的独特和必要的功能，作为鉴定急需的新药的靶点。我们对布氏锥虫RNA编辑连接酶1(TbREL1)和F1-ATPase作为药物靶标的验证(Schnaufer等人，2001年；2005年)，以及高分辨率晶体结构的可用性(邓氏等人，2004年；蒙哥马利等人，2018年)和高通量筛选试验使这些酶成为药物发现工作的有希望和成熟的靶标。所有锥虫都以不寻常的线粒体生物学为特征。例如，它们需要一种独特的RNA编辑形式来表达线粒体基因；这一过程中的一个关键酶是REL1(Schnaufer等人，2001年)。此外，布鲁氏锥虫使用线粒体F1-ATPase不是为了产生ATP，而是作为一种必要的质子泵(Schnaufer等人，2005年)。这些酶在弓形虫、间日弓形虫和布氏锥虫中的同源同源性几乎相同，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-ATPase的小分子抑制剂进行了一些高通量筛选，我们发现并确认了许多结构多样化的低微摩尔IC50值的抑制剂。这个PHD项目将涉及参与进一步的HIT鉴定和Hit-to-Lead开发，特别是在体外(活性分析；使用ITC和表面等离子体共振进行生物物理特性测试；毒理学研究)和体内(针对锥虫和对照细胞)以及通过结晶学、低温电子显微镜和/或计算预测进行目标抑制剂复合体的结构分析。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，惠康细胞生物学中心)，Manfred Auer教授，化学生物学专家(系统生物学中心)，以及Asier Unciti-Broceta教授(爱丁堡癌症研究英国中心)，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养提供了理想的环境。

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.