Discovery of new molecular phenotypes for anti-schistosomal drug screening

抗血吸虫药物筛选新分子表型的发现

基本信息

批准号：
10214505
负责人：
John D Chan
金额：
$ 19.44万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10214505
关键词：
Address Alternative Therapies Animals Anthelmintics Antiparasitic Agents Apoptosis Biological Assay Biology Chemicals Clinical Combined Modality Therapy Data Descriptor Development Disease Drug Combinations Drug Exposure Drug Screening Drug resistance Drug usage Event Exposure to Future Gene Expression Profile Gene Expression Profiling Gene set enrichment analysis Genetic Transcription Genetic Variation Germ Cells Global Change Goals Harvest In Situ Hybridization In Vitro Infection Lead Liver Measurement Measures Mesentery Metabolism Mitosis Molecular Morphology Movement Mus Muscle Nail plate Outcome Parasites Pathway interactions Pharmaceutical Preparations Pharmacotherapy Phenotype Plasmodium Platyhelminths Praziquantel Praziquantel resistance Reporting Research Resistance Schistosoma Schistosomatidae Schistosomiasis Tissues Transcript Treatment Failure Work base combat comparative design differential expression drug action drug discovery drug efficacy drug mechanism druggable target improved in vitro Assay in vivo innovation insight molecular phenotype motor impairment neglect neglected tropical diseases novel novel therapeutics prevent response risk minimization screening transcriptome sequencing

项目摘要

PROJECT SUMMARY: Parasitic Schistosoma blood flukes cause the neglected topical disease schistosomiasis. This disease infects >200 million people but is clinically treated by monotherapy with just one broad spectrum drug, praziquantel (PZQ). The vast scope of the disease and the prospect of PZQ resistance highlight the need for alternative therapies. Indeed, PZQ treatment failure has been reported in the field and resistance can be selected in the lab, indicating that standing genetic variation for resistance already exists. However, drug discovery efforts are hampered by our poor understanding of how existing anti-schistosomal drugs work. Many of these older compounds were discovered >40 years ago using in vivo animal screens. The long-term goal is to identify druggable targets / lead compounds to treat schistosomiasis. The overall objective of this application is to resolve the molecular changes in schistosome biology that underpin the efficacy of existing anti-schistosomal compounds, moving beyond superficial descriptors of worm morphology towards quantitative endpoints that can be assayed for new leads. The central hypothesis is that anthelmintics evoke molecular changes that are more productive screening endpoints than superficial phenotypes. The rationale for this project is that current schistosomiasis drug discovery efforts often focus on in vitro assays for changes in movement or morphology, which are poor predictors of efficacy in vivo. Some anti-schistosomal drugs do cause changes in worm movement / morphology, but others are efficacious in vivo with no effect on movement in vitro, and still other drugs impair movement in vitro but are ineffective in vivo. Better predictors of anti- parasitic action are needed to develop more productive assays. We will pursue two specific aims: (1) Profiling the activity of 10 chemically diverse anti-schistosomal drugs using a panel of molecular assays and (2) Identifying transcriptional signatures of anthelmintics with distinct mechanisms of action. The first aim will resolve mechanistic similarities and differences between these 10 anthelmintics by systematically assessing cellular and molecular changes in worms exposed to drug in vivo. These outcomes will serve as endpoints for the development of quantitative assays for future screens. The second aim will compare the transcriptional responses of parasites to each drug, providing an unbiased readout of global changes to schistosome biology and allowing drugs to be binned according to putative mechanisms of action. Insight into the comparative mechanisms of anti-schistosomal compounds will allow us to rationally select alternative drugs in the event of PZQ treatment failure. This information will also inform drug-combination strategies to prevent the emergence of drug resistance. The research proposed in this application is significant because it will provide new assays for future drug discovery efforts that are better predictors of in vivo anti-parasitic efficacy than existing superficial phenotypes of worm movement and morphology. This research is innovative because it is will establish mechanism-based assays to identify PZQ-alternatives to combat and prevent drug resistance.

项目摘要：寄生血吸虫血吸虫导致被忽视的局部疾病血吸虫病。这种疾病感染超过 2 亿人，但临床上仅通过一种单一疗法进行治疗广谱药物，吡喹酮（PZQ）。该疾病的广泛范围和 PZQ 抗性的前景强调替代疗法的必要性。事实上，PZQ 治疗失败的报道已经在现场出现，并且可以在实验室中选择抗性，这表明抗性的遗传变异已经存在。然而，由于我们对现有抗血吸虫药物的作用了解甚少，药物发现工作受到了阻碍。药物起作用。许多这些较古老的化合物是 40 年前通过体内动物筛选发现的。这长期目标是确定治疗血吸虫病的药物靶标/先导化合物。总体目标该应用的目的是解决血吸虫生物学中支撑其功效的分子变化现有的抗血吸虫化合物，超越了蠕虫形态的表面描述可以分析新线索的定量终点。中心假设是驱虫药会引起分子变化是比表面表型更有效的筛选终点。理由该项目的特点是，当前的血吸虫病药物发现工作通常侧重于体外测定，以了解血吸虫病的变化运动或形态，它们是体内功效的不良预测因素。一些抗血吸虫药物可以引起蠕虫运动/形态的变化，但其他药物在体内有效，对运动没有影响在体外，还有一些药物会损害体外的运动，但在体内无效。更好地预测抗需要寄生作用来开发更有成效的检测方法。我们将追求两个具体目标：（1）分析使用一组分子分析测定 10 种化学成分不同的抗血吸虫药物的活性，以及 (2) 识别具有不同作用机制的驱虫药的转录特征。第一个目标将通过系统评估来解决这 10 种驱虫药之间的机制相似性和差异体内暴露于药物的蠕虫的细胞和分子变化。这些结果将作为终点未来筛选定量分析的发展。第二个目标将比较转录寄生虫对每种药物的反应，提供血吸虫生物学全球变化的公正读数并允许药物根据假定的作用机制进行分类。比较洞察抗血吸虫化合物的作用机制将使我们能够在发生血吸虫病时合理选择替代药物 PZQ治疗失败。这些信息还将为药物组合策略提供信息，以防止出现的耐药性。本申请中提出的研究意义重大，因为它将提供新的测定方法未来的药物发现工作比现有的药物发现工作能够更好地预测体内抗寄生虫功效蠕虫运动和形态的表面表型。这项研究具有创新性，因为它将建立基于机制的测定法来识别 PZQ 替代品以对抗和预防耐药性。