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

• 批准号: 10039280
• 负责人: John D Chan
• 金额: $ 22.08万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10039280
• 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; Genes; 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

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.

项目概要：寄生血吸虫引起被忽视的局部疾病 血吸虫病这种疾病感染超过2亿人，但临床上仅用一种单一疗法治疗 广谱药物吡喹酮（PZQ）。病害的广泛分布及PZQ抗性的前景 强调替代疗法的必要性。事实上，PZQ治疗失败已在该领域报告， 抗性可以在实验室中选择，这表明抗性的长期遗传变异已经存在。 然而，药物发现的努力受到阻碍，我们对现有的抗肿瘤药物是如何产生的了解不足， 药物起作用。这些较老的化合物中的许多是在> 40年前使用体内动物筛选发现的。的 长期目标是确定治疗血吸虫病的药物靶点/先导化合物。总体目标 这项应用的目的是解决支持药物疗效的染色体生物学中的分子变化。 现有的抗寄生虫体化合物，超越了蠕虫形态学的表面描述， 可以针对新导联进行分析的定量终点。中心假设是驱虫药引起 分子变化是比表面表型更有效的筛选终点。的理由 目前的血吸虫病药物发现工作通常集中在体外测定中， 运动或形态，这是体内功效的不良预测因子。一些抗染色体药物 引起蠕虫运动/形态的变化，但其他药物在体内有效，对运动无影响 还有一些药物在体外损害运动，但在体内无效。更好的抗- 需要寄生作用来开发更有效的测定。我们将追求两个具体目标：（1）分析 使用一组分子测定法测定10种化学上不同的抗β-内酰胺酶体药物的活性，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，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， 为未来的筛查开发定量分析。第二个目的是比较转录 寄生虫对每种药物的反应，为寄生虫生物学的全球变化提供了公正的读数 并允许根据假定的作用机制将药物分组。洞察比较 抗β-内酰胺酶化合物的机制将使我们能够在发生以下情况时合理选择替代药物： PZQ治疗失败。这些信息也将告知药物组合策略，以防止出现 抗药性的证据。本申请中提出的研究具有重要意义，因为它将提供新的分析方法 对于未来的药物发现工作，其是比现有的更好的体内抗寄生虫功效的预测因子， 蠕虫运动和形态的表面表型。这项研究是创新的，因为它是将 建立基于机制的分析，以确定PZQ替代品，以对抗和预防耐药性。