Raman spectroscopy uncovers biological changes in extracellular vesicles generated by thyroid tumor cell-fibroblast interplay

M. B. GILARDONI; G. A. BORIOLI; DRUETTA ESTEBAN; LACCONI GABRIELA; M. MONTESINOS; C. G. PELLIZAS; A. C. DONADIO

Mar del Plata

Congreso; Reunión Conjunta SAIC (Sociedad Argentina de Investigacion Clínica), SAI ( Sociedad Argentina de Inmunología) & FAIC (3er Congreso Franco-Argentino de Inmunología), SAFIS (Sociedad Argentina de Fisiología); 2022

SAIC (Sociedad Argentina de Investigacion Clínica), SAI ( Sociedad Argentina de Inmunología), SAFIS (Sociedad Argentina de Fisiología)

Extracellular vesicles (EVs) are a heterogeneous population ofnanoparticles involved in cell-cell communication. Carcinoma is acomplex society of interacting cells, with tumor-stroma crosstalkleading to tumor progression. We demonstrated that thyroid tumorcell-fibroblast (Fb) interaction induces the secretion of metallopro-teinases (MMPs) to culture supernatants (CMs) and promotes amigratory phenotype in the tumor cells. Thyroid tumor cell-Fb cross-talk liberates EVs that are involved in extracellular matrix remod-elling. Raman spectroscopy is a well-established label-free andnon-destructive method for determining chemical characteristics inbiological samples. We utilized Raman Confocal Microspectroscopy(Raman-CM) to explore individual secreted-EVs and to describe theimpact of tumor-stroma interplay in their chemical make-up. Thy-roid tumor cells (TPC-1) or thyroid non-tumor cells (NThyOri) were co-cultured with normal human Fb as a simulation of the thyroidtumor microenvironment (TM). EVs obtained by ultracentrifugationof thyroid cell, Fb and thyroid cell-Fb CMs were characterised byRaman-CM. The Raman spectra of the EVs secreted by thyroidcells either isolated or co-cultured with Fb were compared in orderto distinguish biochemical patterns and their association with EVspotential functionality.Our results show a differential spectral pattern between EVs ob-tained from isolated and co-cultured TPC-1 cells. Characteristicpatterns in the 1000?1550 cm-1 and 2150-2700 cm-1 of the Ramanspectra of EVs from isolated TPC-1 cells are absent in EVs fromco-cultured TPC-1 cells. A set of vibrational signals in the region2800-3000 cm-1 only appear in Fb-TPC-1 derived EVs. There are nochanges in the spectra of EVs originated by isolated or co-culturedNThyOri cells.Taken together these findings suggest an important role of Fbs inEV-phenotype from thyroid tumor. They also open a new approachto study the thyroid TM and its role in thyroid tumor progression.