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

TY - JOUR

T1 - Live-imaging rate-of-kill compound profiling for Chagas disease drug discovery with a new automated high-content assay

AU - Svensen, Nina

AU - Wyllie, Susan

AU - Gray, David W.

AU - De Rycker, Manu

N1 - Funding: This work was funded by Wellcome Trust (https://wellcome.org/) awards 203134/Z/16/Z (NS, DWG), 204672/Z/16/Z (MDR, DWG) and 218448/Z/19/Z (SW). The funder did not play a role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

PY - 2021/10/11

Y1 - 2021/10/11

N2 - Chagas disease, caused by the protozoan intracellular parasite Trypanosoma 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, both in vitro and in vivo, that provide improved understanding of the causes for clinical trial failures. Amongst these are in 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 intracellular T. 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 partners for combination therapy.

AB - Chagas disease, caused by the protozoan intracellular parasite Trypanosoma 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, both in vitro and in vivo, that provide improved understanding of the causes for clinical trial failures. Amongst these are in 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 intracellular T. 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 partners for combination therapy.

UR - https://www.scopus.com/pages/publications/85119290082

U2 - 10.1371/journal.pntd.0009870

DO - 10.1371/journal.pntd.0009870

M3 - Article

C2 - 34634052

SN - 1935-2727

VL - 15

JO - PLoS Neglected Tropical Diseases

JF - PLoS Neglected Tropical Diseases

IS - 10

M1 - e0009870

ER -