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

• Published in: STAR protocols 2023-03, Vol.4 (1), p.102058-102058, Article 102058
• Main Authors: Guerra-Slompo, Eloise Pavão, Picchi-Constante, Gisele Fernanda Assine, Marek, Martin, Romier, Christophe, Sippl, Wolfgang, Zanchin, Nilson Ivo Tonin
• Format: Article
• Language: English
• Subjects: Cell Biology; Cell-based Assays; High-throughput Screening; Microbiology; Model Organisms; Protocol
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2023.102058

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 IC50, CC50, 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 [Display omitted] •In cellulo infection assays for effective compound testing against T. cruzi•High-content image analysis to determine selectivity and toxicity indexes•Bioluminescence in vivo infection assays to evaluate compound efficiency Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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 IC50, CC50, 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.