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

蠕虫感染是存在贫困和发展问题的国家的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其长期可持续增长。发现新的抗蠕虫药物的主要瓶颈之一是缺乏一种快速、定量和重复性的测定方法来筛选和表征成虫形式的候选分子的活性。将在巴西建立和传播新的和更好的研究能力，最大限度地扩大其对贫困和经济增长问题的影响。高含量筛选平台（伦敦和里约）将根据新开发的表型筛选策略，鉴定对成虫有活性的新型小化合物。里约热内卢的研究小组正在开发一种新的平台，用于无偏见地量化药物对蠕虫的作用，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方法的定量图像分析。LSHTM（伦敦，英国）的Bickle小组已经开发了基于血吸虫幼虫生命阶段的类似系统，并且能够在测定开发和图像分析方面提供技术专长。与LSHTM分享想法和经验将大大加快成人染色体异常自动药物筛查平台的开发和整合。LSHTM的Dr. Cristham的小组将与FIOCRUZ小组一起开发一种计算方法，以优先考虑血吸虫属物种（特别是S。mansoni）基因组，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。目标数据的缺乏阻碍了抗血吸虫病活性化合物进一步开发成显示更高效力、更好安全性和降低产生耐药性倾向的药物。为了规避这一限制，将具有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药物机制对化合物进行分类。同时，将通过计算评估具有已知分子靶点并已用于临床治疗其他疾病的药物，以评估其是否适合重新用于治疗帕金森病。牛津蛋白生产设施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.具有作为药物靶点潜力的mansoni蛋白，或者是我们在FIOCRUZ的小组先前鉴定的，或者是从LSHTM.4的Dr. Rehham的小组在[2]中开发的生物信息学管道中选择的。S.将使用OPPF-UK中的高通量自动化流水线筛选[3]中生产的曼氏蛋白以进行结晶。Dr. Rehham和Dr. Silva的团队将合作解决靶蛋白的结构，然后采用基于结构的设计方法来优化药物结合亲和力。将选择最佳化合物进行化学衍生化和结构多样化，以探索结构-活性关系（SAR）。费雷拉博士在UFRJ的小组将合成这些分子。6.巴西的工作坊。将在FIOCRUZ举办两次研讨会，每学期一次。第一个研讨会的主题是高通量蛋白质表达和结晶。第二个研讨会将讨论计算生物学在大数据集目标识别中的应用。

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.