In cellulo and in vivo assays for compound testing against Trypanosoma cruzi

Summary

Here, we describe a combined in cellulo and in vivo approach to identify compounds with higher potential for efficient inhibition of Trypanosoma cruzi. Phase I of in cellulo assays is designed to exclude inactive or toxic compounds, while phase II is designed for accurate IC ₅₀, CC ₅₀, and selective index (SI) determination. Compounds showing high SI are tested using in vivo infection models in parallel with benznidazole to assess their efficacy relative to a reference drug used for Chagas disease treatment.

For complete details on the use and execution of this protocol, please refer to Marek et al. (2021). ¹

Graphical abstract

Highlights

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  In cellulo infection assays for effective compound testing against T. cruzi

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  High-content image analysis to determine selectivity and toxicity indexes

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  Bioluminescence in vivo infection assays to evaluate compound efficiency

Abstract

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Abstract

Here, we describe a combined in cellulo and in vivo approach to identify compounds with higher potential for efficient inhibition of Trypanosoma cruzi. Phase I of in cellulo assays is designed to exclude inactive or toxic compounds, while phase II is designed for accurate IC ₅₀, CC ₅₀, and selective index (SI) determination. Compounds showing high SI are tested using in vivo infection models in parallel with benznidazole to assess their efficacy relative to a reference drug used for Chagas disease treatment.