Live-imaging rate-of-kill compound profiling for Chagas disease drug discovery with a new automated high-content assay

by Nina Svensen, Susan Wyllie, David W. Gray, Manu De Rycker Chagas disease, caused by the protozoan intracellular parasiteTrypanosoma cruzi, is a highly neglected tropical disease, causing significant morbidity and mortality in central and south America. Current treatments are inadequate, and recent clinical trials of drugs inhibiting CYP51 have failed, exposing a lack of understanding of how to translate laboratory findings to the clinic. Following these failures many new model systems have been developed, bothin vitro andin vivo, that provide improved understanding of the causes for clinical trial failures. Amongst these arein vitro rate-of-kill (RoK) assays that reveal how fast compounds kill intracellular parasites. Such assays have shown clear distinctions between the compounds that failed in clinical trials and the standard of care. However, the published RoK assays have some key drawbacks, including low time-resolution and inability to track the same cell population over time. Here, we present a new, live-imaging RoK assay for intracellularT.cruzi that overcomes these issues. We show that the assay is highly reproducible and report high time-resolution RoK data for key clinical compounds as well as new chemical entities. The data generated by this assay allow fast acting compounds to be prioritised for progression, the fate of individual parasites to be tracked, shifts of mode-of-action within series to be monitored, better PKPD modelling and selection of suitable pa...
Source: PLoS Neglected Tropical Diseases - Category: Tropical Medicine Authors: Source Type: research