A ausência da proteína P21 em Trypanosoma cruzi impacta a invasão celular por tripomastigotas e modula a multiplicação e a expressão gênica de amastigotas intracelular de cepas filogeneticamente distintas

Abstract

Trypanosoma cruzi is a protozoan parasite and the causative agent of Chagas disease, a neglected tropical disease with wide distribution in the Americas. This parasite displays high genetic variability, classified into Discrete Typing Units (DTUs), which are associated with biological, clinical, and molecular differences. Among the various factors involved in T. cruzi pathogenicity, the P21 protein stands out as a secreted glycoprotein implicated in modulating the inflammatory response, promoting host cell adhesion and invasion, and potentially regulating the parasite cell cycle. This study aimed to investigate the functional role of the P21 protein through targeted gene knockout in different T. cruzi strains using the CRISPR/Cas9 genome editing system, with a special focus on transcriptomic changes resulting from its absence. Knockout lines were generated in Y (TcII) and G (TcI) strains, representing distinct DTUs. The phenotypic impact of the knockout was evaluated in different developmental forms of the parasite, followed by global gene expression analysis using RNA-seq. Transcriptomic profiling revealed distinct expression patterns between knockout strains, with differentially expressed genes related to key processes such as cell adhesion, signal transduction, stress response, post-translational modifications, and the expression of surface proteins involved in virulence. These findings suggest that P21 plays a modulatory role in T. cruzi biology, with strain-specific effects on host-parasite interactions. The results contribute to a better understanding of P21 function during the parasite life cycle and highlight the utility of genome editing as a powerful tool for functional studies in trypanosomatids.