title: An image-based drug susceptibility assay targeting the placental sequestration of Plasmodium falciparum-infected erythrocytes.
abstract: Placental malaria is a significant cause of all malaria-related deaths globally for which no drugs have been developed to specifically disrupt its pathogenesis. To facilitate the discovery of antimalarial drugs targeting the cytoadherence process of Plasmodium-infected erythrocytes in the placenta microvasculature, we have developed an automated image-based assay for high-throughput screening for potent cytoadherence inhibitors in vitro. Parasitized erythrocytes were drug-treated for 24 h and then allowed to adhere on a monolayer of placental BeWo cells prior to red blood cell staining with glycophorin A antibodies. Upon image-acquisition, drug effects were quantified as the proportion of treated parasitized erythrocytes to BeWo cells compared to the binding of untreated iRBCs. We confirmed the reliability of this new assay by comparing the binding ratios of CSA- and CD36-panned parasites on the placental BeWo cells, and by quantifying the effects of chondroitin sulfate A, brefeldin A, and artemisinin on the binding. By simultaneously examining the drug effects on parasite viability, we could discriminate between cytoadherence-specific inhibitors and other schizonticidal compounds. Taken together, our data establish that the developed assay is highly suitable for drug studies targeting placental malaria, and will facilitate the discovery and rapid development of new therapies against malaria.
authors.affiliation: Center for Neglected Diseases Drug Discovery (CND3), Institut Pasteur Korea, Seongnam-si, Gyeonggi-do, South Korea.
authors.forename: Min-Je
authors.initials: MJ
authors.lastname: Ku
authors.forename: Fernando de M
authors.initials: Fde M
authors.lastname: Dossin
authors.forename: Michael A E
authors.initials: MA
authors.lastname: Hansen
authors.forename: Auguste
authors.initials: A
authors.lastname: Genovesio
authors.forename: Lawrence
authors.initials: L
authors.lastname: Ayong
authors.forename: Lucio H
authors.initials: LH
authors.lastname: Freitas-Junior
mesh_terms: D000465:Algorithms; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D037621:Artemisinins / Q000494:pharmacology; D001331:Automation; D020126:Brefeldin A / Q000494:pharmacology; D018955:CD36 Antigens / Q000096:biosynthesis; D002448:Cell Adhesion; D002460:Cell Line; D002470:Cell Survival; D002809:Chondroitin Sulfates / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical; D004912:Erythrocytes / Q000469:parasitology*; D005260:Female; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D016778:Malaria, Falciparum / Q000188:drug therapy* / Q000469:parasitology*; D010920:Placenta / Q000469:parasitology*; D010963:Plasmodium falciparum / Q000378:metabolism*; D011247:Pregnancy
chemical_list: D000962:Antimalarials; D037621:Artemisinins; D018955:CD36 Antigens; D020126:Brefeldin A; D002809:Chondroitin Sulfates; C031327:artemisinin