Identifying key amino acid metabolic steps as potential drug targets for Chagas disease treatment
Wellcome Trust
University of Sao Paulo
Trypanosoma cruzi, the causal agent of Chagas disease, has a complex digenetic life cycle. This organism is well adapted to handle environmental changes, presenting high metabolic flexibility. In this context, amino acids and their metabolism play crucial roles in cell differentiation, maintenance of cell volume and response to different types of stress. This proposal aims to identify the key steps of the amino acid metabolic network which are crucial for T. cruzi to complete its life cycle. With this purpose, we will use the bar-seq CRISPR-Cas9 genome editing strategy to generate mutant knockout cell lines for the coding sequences of 40 enzymes putatively involved in the metabolism of amino acids. Each cell line will be identified by a unique sequence barcode and they will be pooled to allow parallel phenotyping during cell differentiation and infection, both in vitro and in vivo. Relative fitness will be assessed by next-generation DNA sequencing. Identification of the enzymes that are essential for parasite development and infectivity will provide insights on the metabolic strategies T. cruzi evolved to adapt to diverse environments as well as novel targets for target-specific drug design and ligand structure-based screenings in the future.
2.1 Biological and endogenous factors