RAMAN SPECTROSCOPY UNCOVERS BIOLOGICAL CHANGES IN EXTRACELLULAR VESICLES GENERATED BY THYROID TUMOR CELL-FIBROBLAST INTERPLAY

Mar del Plata

Congreso; REUNION ANUAL DE SOCIEDADES DE BIOCIENCIAS- SAIC-SAI-SAFIS-; 2022

Extracellular vesicles (EVs) are a heterogeneous population of nanoparticles involved in cell-cell communication. Carcinoma is a complex society of interacting cells, with tumor-stroma crosstalk leading to tumor progression.We demonstrated that thyroid tumor cell-fibroblast (Fb) interaction induces the secretion of metalloproteinases (MMPs) to culture supernatants (CMs) and promotes a migratory phenotype in the tumor cells. Thyroid tumor cell-Fb crosstalk liberates EVs that are involved in extracellular matrix remodelling. Raman spectroscopy is a well-established label-free and non-destructive method for determining chemical characteristics in biological samples. We utilized Raman Confocal Microspectroscopy (Raman-CM) to explore individual secreted-EVs and to describe the impact of tumor-stroma interplay in their chemical make-up. Thyroid tumor cells (TPC-1) or thyroid non-tumor cells (NThyOri) were co-cultured with normal human Fb as a simulation of the thyroid tumor microenvironment (TM). EVs obtained by ultracentrifugation of thyroid cell, Fb and thyroid cell-Fb CMs were characterised by Raman-CM. The Raman spectra of the EVs secreted by thyroid cells either isolated or co-cultured with Fb were compared in order to distinguish biochemical patterns and their association with EVs potential functionality.Our results show a differential spectral pattern between EVs obtained from isolated and co-cultured TPC-1 cells. Characteristic patterns in the 1000?1550 cm-1 and 2150-2700 cm-1 of the Raman spectra of EVs from isolated TPC-1 cells are absent in EVs from co-cultured TPC-1 cells. A set of vibrational signals in the region 2800-3000 cm-1 only appear in Fb-TPC-1 derived EVs. There are no changes in the spectra of EVs originated by isolated or co-cultured NThyOri cells. Taken together these findings suggest an important role of Fbs in EV-phenotype from thyroid tumor. They also open a new approach to study the thyroid TM and its role in thyroid tumor progression.