New001 Building research capacity for schistosomiasis drug discovery & development through high-content imaging & structural molecular biology studies

在有贫穷和发展问题的国家，蠕虫感染是一个巨大的公共卫生问题，在这些国家，疾病发病率造成的经济影响最终阻碍了它们的长期可持续增长。发现新的抗蠕虫药物的主要瓶颈之一是缺乏一种快速、定量和繁殖的检测方法来筛选和表征候选分子在成虫形式中的活性。将进行的研究活动简述如下。将在巴西建立和传播新的和改进的研究能力，最大限度地发挥它们对贫困和经济增长问题的影响。high Content Screen平台（London和里约热内卢）将基于新开发的表型筛选策略，鉴定出对成年血吸虫具有活性的新型小化合物。巴西里约热内卢的研究小组正在开发一种新的平台，用于对针对蠕虫的药物作用进行无偏量化，该平台基于未标记的成年寄生虫的自动成像和随后使用定制开发的方法进行定量图像分析。LSHTM （London， UK）的Bickle小组已经开发了一个基于血吸虫幼虫生命阶段的类似系统，并且能够在分析开发和图像分析方面提供技术专长。与LSHTM的经验分享将极大地促进成虫药物筛选自动化平台的发展和巩固。furham博士在LSHTM的团队将与FIOCRUZ团队一起开发一种计算方法，优先考虑血吸虫物种（特别是曼氏血吸虫）基因组中编码的药物靶点，并评估目前临床使用的药物是否可以重新用于治疗血吸虫病。缺乏目标数据阻碍了进一步将抗血吸虫病的活性化合物开发成效力更高、安全性更好和产生耐药性倾向更低的药物。为了规避这一限制，将未知机制的化合物诱导的表型与已知药物产生的表型进行比较，然后使用统计模型根据已知的药物机制对化合物进行分类。同时，已知分子靶点并已用于临床治疗其他疾病的药物将进行计算评估，以评估其重新用于治疗血吸虫病的适用性。牛津蛋白质生产工厂开发了一系列高度专业化的技术，包括机器人系统，以实现重组蛋白的高通量表达、纯化和结晶。该平台将应用于重组S. mansoni蛋白的生产，这些重组S. mansoni蛋白具有作为药物靶点的潜力，可以是我们的团队在FIOCRUZ之前确定的，也可以是从lshtm的Furnham博士的团队在[2]开发的生物信息学管道中选择的。在[3]中生产的S. mansoni蛋白将使用OPPF-UK的高通量自动化管道进行筛选结晶。弗恩汉姆博士和席尔瓦博士的团队将合作解决目标蛋白的结构，然后采用基于结构的设计方法来优化药物结合亲和力。选择最佳化合物进行化学衍生和结构多样化，探索构效关系（SAR）。费雷拉博士在UFRJ的团队将合成这些分子。6. 在巴西的工作坊。FIOCRUZ将举办两次研讨会，每学期一次。第一次研讨会将以高通量蛋白表达和结晶为主题。第二场研讨会将讨论计算生物学在大数据目标识别中的应用。

Helminth worm infections are a huge public health problem in countries with poverty and development issues where the economic impacts due to disease morbidity ultimately hamper their long-term sustainable growth. One of the main bottlenecks in the discovery of new anti-helminthic drugs is the lack of a fast, quantitative and reproductive assay method to screen and characterize the activity of candidate molecules in adult worm forms.The research activities to be carried out are briefly described below. New and improved research capacities will be built and disseminated in Brazil, maximising their impact on the issues of poverty and economic growth.1. The Hight Content Screen platforms (London and Rio) will identify novel small compounds active against adult schistosome worms based on a newly developed phenotypic screening strategy. The group in Rio de Janeiro is developing a novel platform for unbiased quantification of drug action against helminths that is based on automated imaging of unlabelled adult parasite worms and subsequent quantitative image analysis using custom-developed methods. A similar system based on the larval life stage of Schistosoma has been developed by the Bickle group at LSHTM (London, UK) and are able to lend technical expertise in both assay development and image analysis. Sharing ideas and experience with LSHTM would greatly accelerate the development and consolidation of the automated drug-screening platform for adult schistosomes.2. Dr. Furnham's group at LSHTM together with the FIOCRUZ group will develop a computational method to prioritize drug targets encoded within the genomes of Schistosoma species (in particular S. mansoni) genomes and to assess if drugs currently in clinical use can be repurposed to treat schistosomiasis. Deficiency in target data prevents further development of active compounds against schistosomiasis into drugs showing higher potency, better safety and reduced propensity to develop resistance. To circumvent this limitation, phenotypes induced by compounds with unknown mechanism will be compared to the ones produced by known drugs and a statistical model will then be used to classify the compounds according to known drug mechanisms. Concurrently, drugs with known molecular targets and already used in clinical use for other diseases will be computationally evaluated to assess their suitability to be repurposed to treat schistosomiasis.3. The Oxford Protein Production Facility has developed a range of highly specialized technologies incorporating robotic systems to enable the high throughput expression, purification and crystallization of recombinant proteins. This platform will be applied to the production of recombinant S. mansoni proteins with potential as drug targets either previously identified by our group at FIOCRUZ or selected from bioinformatics pipelines developed in [2] by Dr. Furnham's group at LSHTM.4. S. mansoni proteins that have been produced in [3] will be screened for crystallization using the high throughput automated pipeline in the OPPF-UK. Dr. Furnham's and Dr. Silva's groups will collaborate to solve the structures of the target proteins, and then employ structure-based design methods to optimize drug binding affinities.5. The best compounds will be selected for chemical derivatization and structural diversification to explore structure-activity relationships (SAR). Dr. Ferreira's group in UFRJ will synthesize the molecules. 6. Workshops in Brazil. Two workshops will be held at FIOCRUZ, one in each semester. The first workshop will have as its theme the high-throughput protein expression and crystallization. The second workshop will be on the Application of Computational Biology for Target Identification from Big Datasets.