A panel of phenotypically and genotypically diverse bioluminescent: fluorescent Trypanosoma cruzi strains as a resource for Chagas disease research

Abstract

Chagas disease is caused by Trypanosoma cruzi, a protozoan parasite that displays considerable genetic diversity. Infections result in a range of pathological outcomes, and different strains can exhibit a wide spectrum of anti-parasitic drug tolerance. The genetic determinants of infectivity, virulence and therapeutic susceptibility remain largely unknown. As experimental tools to address these issues, we have generated a panel of bioluminescent:fluorescent parasite strains that cover the diversity of the T. cruzi species. These reporters allow spatio-temporal infection dynamics in murine models to be monitored in a non-invasive manner by in vivo imaging, provide a capability to detect rare infection foci at single-cell resolution, and represent a valuable resource for investigating virulence and host:parasite interactions at a mechanistic level. Importantly, these parasite reporter strains can also contribute to the Chagas disease drug screening cascade by ensuring that candidate compounds have pan-species in vivo activity prior to being advanced into clinical testing. The parasite strains described in this paper are available on request. AUTHOR SUMMARYChagas disease results from infection with the protozoan parasite Trypanosoma cruzi and is a major public health problem throughout Latin America. T. cruzi is a genetically diverse species and infection can result in a wide range of pathological outcomes, mainly associated with the heart and/or digestive tract. Research on Chagas disease, ranging from fundamental biology to drug development, has been greatly aided by the availability of genetically modified parasite reporter strains that express bioluminescent:fluorescent fusion proteins. In combination with mouse models and imaging technology, these strains allow infections to be monitored in real-time, with high sensitivity, and infection foci to be visualised at single-cell resolution. Here, we describe an extensive panel of bioluminescent and fluorescent strains that cover the diversity of the T. cruzi species. These reporter strains, that are available on request, should have wide utility in many areas of Chagas disease research. In particular, as part of the drug development screening programme, they can be used to ensure that candidate compounds have in vivo activity across the species prior to being advanced into clinical testing.