Discovery by organism based high-throughput screening of new multi-stage compounds affecting Schistosoma mansoni viability, egg formation and production

Abstract

Schistosomiasis, one of the most prevalent neglected parasitic diseases affecting humans and animals, is caused by the Platyhelminthes of the genus Schistosoma. Schistosomes are the only trematodes to have evolved sexual dimorphism and the constant pairing with a male is essential for the sexual maturation of the female. Pairing is required for the full development of the two major female organs, ovary and vitellarium that are involved in the production of different cell types such as oocytes and vitellocytes, which represent the core elements of the whole egg machinery. Sexually mature females can produce a large number of eggs each day. Due to the importance of egg production for both life cycle and pathogenesis, there is significant interest in the search for new strategies and compounds not only affecting parasite viability but also egg production. Here we use a recently developed high-throughput organism-based approach, based on ATP quantitation in the schistosomula larval stage of Schistosoma mansoni for the screening of a large compound library, and describe a pharmacophore-based drug selection approach and phenotypic analyses to identify novel multi-stage schistosomicidal compounds. Interestingly, worm pairs treated with seven of the eight compounds identified show a phenotype characterized by defects in eggshell assemblage within the ootype and egg formation with degenerated oocytes and vitelline cells engulfment in the uterus and/or oviduct. We describe promising new molecules that not only impair the schistosomula larval stage but also impact juvenile and adult worm viability and egg formation and production in vitro.

Author summary

Schistosomiasis is a neglected disease caused by parasitic flatworms called schistosomes. The disease affects hundreds of millions of people in developing countries in the poorest tropical and subtropical regions of the world and it represents a major public health and socio-economical problem in several countries. In humans, these blood flukes reside in the mesenteric and vesicle venules. They have a life span of many years and produce hundreds of eggs daily, which are able to pass through the gut lumen or the bladder to be finally excreted into the environment for maintaining the life cycle. Part of the eggs can be trapped in host tissues inducing immunologically mediated granulomatous inflammation and fibrosis leading eventually to severe sequelae such as hepatosplenomegaly and even death. Importantly, schistosome infections increase susceptibility to other parasitic, bacterial and viral diseases. To date, essentially a single drug, praziquantel, is available to treat this parasitic disease. Despite its high tolerability and efficacy against adult parasites it has an incomplete efficacy across all stages of the S. mansoni life cycle and it does not prevent reinfection. Moreover the potential risk of drug resistance is an increasing concern. In search of novel schistosomicidal molecules we screened a large compound collection using the schistosomula, larval stage of the parasite. We identified eight novel molecules able to impair viability of schistosomula, juvenile and adult worms and also egg formation and production, two important features required for both disease transmission and progression.