Development of a High-Content Stem Cell–Focused Drug Screening Platform for Schistosoma mansoni Using Developmentally Advanced Liver Schistosomula: Functional Insights and New Protein Target Leads
📄 英文摘要 English Abstract
BACKGROUND: Globally, more than a quarter of a billion people are infected with Schistosoma blood parasites that cause the debilitating disease human schistosomiasis. To assist drug discovery, we developed a novel high content drug screening platform that exploits the extraordinary stem cell biology of the developing parasite. METHOD: Employing non-synchronous in vitro-grown liver stage schistosomula, fluorescence-based staining, confocal laser scanning microscopy, and segmentation of multi-parasite images, the platform was designed to enable high-content quantitative analysis of dividing somatic neoblasts within the parasite to define phenotype. RESULTS: To establish, refine, and test the approach we analysed stem cell proliferation in liver stage schistosomula and validated the impact of EC-144, a compound that supresses Schistosoma neoblast proliferation. Primary drug screening was then done using a stem cell focused library containing 280 compounds, 45 of which attenuated stem cell proliferation by at least 75%. Sixteen 'hit' compounds were prioritised for further testing/confirmation, with testing done in both simple and complex media. Six compounds were finally selected for further investigation, all of which killed or constrained the development of schistosomula and abolished both somatic and germinal stem cell proliferation in adult male and female worms. CONCLUSIONS: In addition to identifying a panel of new anti-schistosomal compounds with associated predicted targets, the findings provide mechanistic insights into the somatic stem cell biology of schistosomes. Further application of this platform, which offers a new approach to assessing compound susceptibility on schistosomula, should accelerate lead compound discovery to help fight human schistosomiasis.
📄 中文摘要 Chinese Abstract
📋 英文结构化总结 English Structured Summary
摘要整理
Background:
Globally, more than a quarter of a billion people are infected with Schistosoma blood parasites that cause the debilitating disease human schistosomiasis. To assist drug discovery, a novel high content drug screening platform was developed that exploits the extraordinary stem cell biology of the developing parasite.
Methods:
The platform employed non-synchronous in vitro-grown liver stage schistosomula, fluorescence-based staining, confocal laser scanning microscopy, and segmentation of multi-parasite images. It was designed to enable high-content quantitative analysis of dividing somatic neoblasts within the parasite to define phenotype.
Results:
The approach was established, refined, and tested by analysing stem cell proliferation in liver stage schistosomula and validating the impact of EC-144, a compound that supresses Schistosoma neoblast proliferation. Primary drug screening used a stem cell focused library containing 280 compounds, 45 of which attenuated stem cell proliferation by at least 75%. Sixteen 'hit' compounds were prioritised for further testing/confirmation in both simple and complex media, and six compounds were finally selected for further investigation; all killed or constrained the development of schistosomula and abolished both somatic and germinal stem cell proliferation in adult male and female worms.
Data Summary:
More than a quarter of a billion people are infected globally. The primary screen contained 280 compounds, 45 attenuated stem cell proliferation by at least 75%, 16 'hit' compounds were prioritised, and 6 compounds were finally selected. The six selected compounds killed or constrained schistosomula development and abolished both somatic and germinal stem cell proliferation in adult male and female worms.
Conclusions:
The findings identified a panel of new anti-schistosomal compounds with associated predicted targets and provided mechanistic insights into the somatic stem cell biology of schistosomes. Further application of the platform, which offers a new approach to assessing compound susceptibility on schistosomula, should accelerate lead compound discovery to help fight human schistosomiasis.
Practical Significance:
The platform provides a new approach to assessing compound susceptibility on schistosomula and can accelerate lead compound discovery to help fight human schistosomiasis.
📋 中文结构化总结 Chinese Structured Summary
背景:
全球有超过2.5亿人感染血吸虫（Schistosoma）寄生虫，该寄生虫可引起致残性的人血吸虫病。为辅助药物发现，研究者开发了一种新型高内涵药物筛选平台，该平台利用发育期寄生虫非凡的干细胞生物学特性。
方法:
该平台采用非同步体外培养的肝期童虫、基于荧光的染色、激光共聚焦显微镜以及多寄生虫图像分割。其设计旨在实现对寄生虫内分裂的体细胞新芽细胞进行高内涵定量分析，以界定表型。
结果:
该方法通过分析肝期童虫中的干细胞增殖，并验证EC-144（一种抑制血吸虫新芽细胞增殖的化合物）的影响，得以建立、优化和测试。初级药物筛选使用了一个以干细胞为重点的化合物库，包含280种化合物，其中45种使干细胞增殖至少降低75%。16种“命中”化合物被优先在简单培养基和复杂培养基中进行进一步测试/确认，最终有6种化合物被选定用于进一步研究；这些化合物均杀死或限制了童虫的发育，并消除了成年雄性和雌性虫体中体细胞和生殖细胞干细胞的增殖。
数据摘要:
全球有超过2.5亿人感染。初筛包含280种化合物，其中45种使干细胞增殖至少降低75%，16种“命中”化合物被优先处理，最终选定6种化合物。这6种选定化合物杀死或限制了童虫发育，并消除了成年雄性和雌性虫体中体细胞和生殖细胞干细胞的增殖。
结论:
研究结果鉴定出一组新的抗血吸虫化合物及其相关预测靶点，并为血吸虫体细胞干细胞生物学提供了机制层面的见解。该平台的进一步应用可提供一种评估化合物对童虫敏感性的新方法，应有助于加速先导化合物发现，以对抗人血吸虫病。
实践意义:
该平台提供了一种评估化合物对童虫敏感性的新方法，并可加速先导化合物发现，以帮助对抗人血吸虫病。