title: Development and validation of a high-throughput anti-Wolbachia whole-cell screen: a route to macrofilaricidal drugs against onchocerciasis and lymphatic filariasis.
abstract: There is an urgent need to develop new, safe, and affordable macrofilaricidal drugs for onchocerciasis and lymphatic filariasis treatment and control. The Anti-Wolbachia Consortium (A·WOL) aims to provide a novel treatment with macrofilaricidal activity by targeting the essential bacterial symbiont Wolbachia. The consortium is currently screening a diverse range of compounds to find new chemical space to drive this drug discovery initiative and address this unmet demand. To increase the throughput and capacity of the A·WOL cell-based screen, we have developed a 384-well format assay using a high-content imaging system (Operetta) in conjunction with optimized Wolbachia growth dynamics in the C6/36 Aedes albopictus mosquito cell line. This assay uses texture analysis of cells stained with SYTO 11 as a direct measure of bacterial load. This validated assay has dramatically increased the capacity and throughput of the A·WOL compound library screening program 25-fold, enriching the number of new anti-Wolbachia hits identified for further development as potential macrofilaricides for onchocerciasis and lymphatic filariasis.
authors.affiliation: Department of Parasitology, Liverpool School of Tropical Medicine, Liverpool, UK.
authors.forename: Rachel H
authors.initials: RH
authors.lastname: Clare
authors.affiliation: Department of Parasitology, Liverpool School of Tropical Medicine, Liverpool, UK.
authors.forename: Darren A N
authors.initials: DA
authors.lastname: Cook
authors.affiliation: Department of Parasitology, Liverpool School of Tropical Medicine, Liverpool, UK.
authors.forename: Kelly L
authors.initials: KL
authors.lastname: Johnston
authors.affiliation: Department of Parasitology, Liverpool School of Tropical Medicine, Liverpool, UK.
authors.forename: Louise
authors.initials: L
authors.lastname: Ford
authors.affiliation: Department of Parasitology, Liverpool School of Tropical Medicine, Liverpool, UK.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Ward
authors.affiliation: Department of Parasitology, Liverpool School of Tropical Medicine, Liverpool, UK Mark.Taylor@lstmed.ac.uk.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Taylor
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology* / Q000627:therapeutic use; D000969:Antinematodal Agents / Q000494:pharmacology* / Q000627:therapeutic use; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery / Q000379:methods* / Q000592:standards; D004605:Elephantiasis, Filarial / Q000188:drug therapy; D057166:High-Throughput Screening Assays / Q000379:methods* / Q000592:standards; D006801:Humans; D058069:Neglected Diseases / Q000188:drug therapy; D009854:Onchocerca / Q000382:microbiology; D009855:Onchocerciasis / Q000188:drug therapy; D015203:Reproducibility of Results; D020577:Wolbachia / Q000187:drug effects* / Q000502:physiology*
chemical_list: D000900:Anti-Bacterial Agents; D000969:Antinematodal Agents
keywords: cell-based assays; fluorescence methods; high-content screening; phenotypic drug discovery