Extracellular Vesicles Facilitate the Interaction of Trypanosoma cruzi with the Extracellular Matrix

izadora ^1,2,3, Nusrat Sattar^2,3, Sarah Beatriz de Fucio Barros^2,3, Marcel Ivan Ramirez ^2,3

^1. UEL, Universidade estadual de Londrina ; Rua Pernambuco 540, Londrina, PR, 86020-120
^2. ICC, Instituto Carlos Chagas ; Rua Prof. Algacyr Munhoz Mader 3775, Curitiba, PR, 81350-010 (41) 3316-3230
^3. ufpr, Universidade federal de Parana; Rua XV de Novembro 1299, Curitiba, PR, 80060-000

The extracellular matrix (ECM) plays a central role in host–parasite interactions by regulating adhesion, invasion, and tissue tropism. Trypanosoma cruzi, the causative agent of Chagas disease, dynamically interacts with ECM components during infection. Proteomic analysis revealed that extracellular vesicles (EVs), particularly those derived from Caco-2 cells infected with the Dm28c strain, are enriched in adhesion-related proteins. Based on these findings, we hypothesize that EVs contribute to T. cruzi infection by interacting with ECM components such as laminin and fibronectin. Ongoing experiments aim to evaluate EV affinity and chemotactic responses toward isolated ECM proteins, to identify potential vesicle targets and guide the development of new therapeutic strategies to control chronic infection.

Materials and Methods

Caco-2 intestinal epithelial cells and C2C12 myoblasts were infected for 2 hours with culture-derived trypomastigotes (TCTs) from the CL Brener (TcVI) and Dm28c (TcI) strains. Large EVs (LEVs) were isolated from infected cultures, quantified by BCA assay, and applied (5 µg/well) to Matrigel-coated Transwell inserts (3 µm pore). Parasite migration to the lower chamber was assessed at 1, 2, 3, 4, and 24 hours using a Neubauer chamber. Affinity and chemotaxis assays with purified ECM components are currently underway.

Results

Inserts without ECM allowed TCT migration within 1 hour. When ECM was present but EVs were absent, parasite migration occurred only after 24 hours. The addition of LEVs from both infected and uninfected cells facilitated earlier migration through the ECM, indicating that EVs contribute to enhanced motility or ECM modification.

Conclusion

EVs appear to induce rapid ECM remodeling or signaling changes that promote T. cruzi migration. We are currently investigating the hypothesis that EVs interact directly with matrix components such as laminin or fibronectin, contributing to tissue-specific parasite dissemination. The differential export of ECM-related proteins may reflect adaptive strategies used by the parasite to colonize distinct tissues, supporting the role of EVs as key determinants of infection dynamics and chronic disease progression.

Keywords: Trypanosoma cruzi; extracellular vesicles; extracellular matrix; laminin; fibronectin; host–pathogen interaction; chronic Chagas disease.

Agradecimentos: UFPR,FIOCRUZ ,CNPq and CAPES