Infections with parasitic nematodes are among the most significant of the neglected tropical diseases affecting about a billion people living mainly in tropical regions with low economic activity. The most effective current strategy to control nematode infections involves large scale treatment programs with anthelmintic drugs. This strategy is at risk from the emergence of drug resistant parasites. Parasitic nematodes also affect livestock, which are treated with the same limited group of anthelmintic drugs. Livestock parasites resistant to single drugs, and even multi-drug resistant parasites, are appearing in many areas. There is therefore a pressing need for new anthelmintic drugs. Here we use the nematode Caenorhabditis elegans as a model for parasitic nematodes and demonstrate that sinefungin, a competitive inhibitor of methyltransferases, causes a delay in development and reduced fecundity, and inhibits spliced leader trans-splicing. Spliced leader trans-splicing is an essential step in gene expression that does not occur in the hosts of parasitic nematodes, and is therefore a potential target for new anthelmintic drugs. We have exploited the ability of sinefungin to inhibit spliced leader trans-splicing to adapt a green fluorescent protein based reporter gene assay that monitors spliced leader trans-splicing for high-throughput screening for new anthelmintic compounds. We have established a protocol for robust high-throughput screening, combining mechanical dispensing of living C. elegans into 384- or 1536- well plates with addition of compounds using an acoustic liquid dispenser, and the detection of the inhibition of SL trans-splicing using a microplate reader. We have tested this protocol in a first pilot screen and envisage that this assay will be a valuable tool in the search for new anthelmintic drugs.

中文翻译：

---

一种高通量筛选，用于鉴定通过靶向剪接前导反式剪接抑制线虫基因表达的化合物。


寄生线虫感染是最重要的被忽视的热带疾病之一，影响着主要生活在经济活动较低的热带地区的约十亿人。目前控制线虫感染最有效的策略涉及使用驱虫药物进行大规模治疗计划。由于耐药寄生虫的出现，这一策略面临风险。寄生线虫也会影响牲畜，牲畜也用相同的有限组驱虫药治疗。许多地区都出现了对单一药物耐药的牲畜寄生虫，甚至多重耐药寄生虫。因此，迫切需要新的驱虫药物。在这里，我们使用线虫秀丽隐杆线虫作为寄生线虫模型，并证明 Sinefungin（甲基转移酶的竞争性抑制剂）会导致发育延迟和繁殖力降低，并抑制剪接前导反式剪接。剪接前导反式剪接是基因表达的一个重要步骤，在寄生线虫宿主中不会发生，因此是新型驱虫药物的潜在靶点。我们利用了sinefungin抑制剪接前导反式剪接的能力，以适应基于绿色荧光蛋白的报告基因检测，该检测监测剪接前导反式剪接，以高通量筛选新的驱虫化合物。我们建立了一个稳健的高通量筛选方案，将活体线虫机械分配到 384 或 1536 孔板中，并使用声学液体分配器添加化合物，并使用酶标仪。 我们已经在第一次试点筛选中测试了该方案，并设想该测定将成为寻找新驱虫药物的宝贵工具。