INSTITUTO DE INVESTIGACIONES BIOTECNOLOGICAS
Unidad Ejecutora - UE
congresos y reuniones científicas
Trypanosoma cruzi trypomastigotes migrate across mammalian 3D-spheroids by a paracellular route in a protease-dependent manner.
CAEIRO, L; RODRIGUEZ, ME; MASIP, Y; RIZZI, M; TEKIEL, V.
Congreso; XXXII Reunión Anual Soc Arg Protozool.; 2020
Soc Arg protozool
The mechanisms associated to dissemination and tissue colonization in T. cruzi infection are largely speculative. Transmigration is the capacity of traverse biological barriers enabling pathogens to spread deeper into tissues. The role of transmigration in the biology of T. cruzi is still unknown. This is probably because the extremely simplicity of monolayer cultures to reproduce a tissue-mimicking environment, as well as the extremely complexity of animal models. Three-D cultures could be an excellent alternative because they reproduce the microarchitecture of tissues, providing an environment similar to the encountered during natural infections. In this work, we used 3D-spheroids of Hela-RFP cells to analyze the transmigration ability of T. cruzi trypomastigotes (tryp) of CL Brener (highly virulent) and SylvioX10 (low virulent) strains. In previous studies we determined that CL Brener tryp transmigrate and penetrate deeply inside spheroids, while most SylvioX10 parasites remain on the superficial layers. CL Brener tryp were usually found in between cell-cell contact sites, suggesting that they are using a paracellular migration pathway. Accordingly, disruption of cell-cell contacts (tight and adherent junctions) increased tryp transmigration and invasion. On the other hand, protease inhibitors strongly blocked the transmigration process of CL Brener tryp. Finally, comparative surface proteomes of tryp from CL Brener and SylvioX10 revealed more than 200 proteins exclusively expressed on the membrane of migrant parasites. Among them, we highlight extracellular matrix (ECM) binding proteins (gp85, CRT), proteases (gp63) and flagellar motility-related proteins (FM7, RSP11). Alltogether the results indicate that transmigration of T. cruzi trypomastigotes is a multistep process, which possibly involve (1) the disruption of cell-cell junctions; (2) the degradation of ECM; (3) the interaction with components of ECM and (4) the motility of parasites