Thyroid tumor cell-fibroblasts crosstalk release extracellular vesicles able to promote thyroid tumor progression

ROYO F; MONTESINOS MM; FALCÓN-PEREZ JM; BRAVO MIANA RC; NEGRETTI BORGA DANA; PELLIZAS CG; SOLER MF; RABINOVICH GA; DONADIO AC

Buenos Aires

Workshop; ISEV Workshop "Extracellular Vesicles and Immunology"; 2020

international Society of Extracellular Vesicles

Carcinomas comprise tumor and non-tumor interacting cells. In the tumor microenvironment (TME), cell-cell interactions, soluble molecules and extracellular vesicles (EVs) are involved in tumor-stroma crosstalk. Metalloproteinases (MMPs) and Galectin-1 (Gal-1) are soluble factors that regulate tumor-stroma interplay, promoting cell proliferation, migration and modulation of immune response, enhancing tumor development. In thyroid cancer (TC), the most common endocrine malignancy, Gal-1 and MMPs expression in tumor cells correlates with tumor aggressiveness. However, TME contribution to TC progression is still underexplored. Using a simulation of thyroid-TME, we focused on the role of EVs modifying Fibroblasts (Fb), encouraging tumor promotion. To this, tumor (TPC-1) or non-tumor (NThyOri) thyroid cells were co-cultured with normal Fb. EVs, obtained by ultracentrifugation of thyroid cell, Fb and Fb-thyroid cell culture supernatants (CMs) were characterized by Nanoparticle Tracking Analysis and Western blot. The uptake of EVs by Fb was assayed using transwells, staining the EVs-producing cells with fluorescent lipophilic dyes and sensing the signal by flow cytometry and fluorescence microscopy. MMPs were studied by zymography in EVs, and CMs of EVs-stimulated Fb. Gal-1 expression was analyzed by RT-qPCR in Fb treated with EVs.EVs released by thyroid and Fb-thyroid cells ranged between 90-200nm and expressed the classical exosome markers CD63, CD9 and CD81. No differences are observed among the number of EVs released by Fb, TPC and NThyOri cells or Fb-TPC and Fb-NThyOri co-cultured cells. Fb uptake a significant higher quantity of EVs released from TPC-1 than from NThyOri cells. EVs released from Fb-TPC-1 co-cultured cells significatively increase the activity of MMP2 as well as Gal-1 mRNA expression in Fb. These results suggest an active participation of Fb in the induction of a permissive milieu for thyroid tumor progression, being modulated by thyroid TME-derived EVs.