Extracellular vesicles from thyroid cancer harbor a functional machinery involved in extracellular matrix remodeling

• Published in: European journal of cell biology Vol. 101; no. 3; p. 151254
• Main Authors: Bravo-Miana, Rocío del Carmen, Soler, María Florencia, Ceschin, Danilo Guillermo, Royo, Félix, Negretti-Borga, Dana María, Azkargorta, Mikel, Elortza, Félix, Montesinos, María del Mar, Pellizas, Claudia Gabriela, Falcón-Pérez, Juan Manuel, Donadio, Ana Carolina
• Format: Journal Article
• Language: English
• Published: Elsevier GmbH 01-06-2022; Elsevier
• Subjects: Co-cultures; Extracellular vesicles; Fibroblasts; MMP2; Proteomics; Thyroid cancer; PCoA; RT; TW; CMs; PPI; Extracellular vesicles; TW-CMs; Co-cultures; CAFs; FDR; Fibroblasts; SFM; Fb; EVs; CI; MMP2; MMPs; GO; DAPI; DEqMS; TME; ECM; Thyroid cancer; nLC-MS/MS; Proteomics; FASP; NTA; DEPs; PTC; ORA
• ISSN: 0171-9335; 1618-1298
• DOI: 10.1016/j.ejcb.2022.151254

Extracellular vesicles (EVs) participate in cell-stroma crosstalk within the tumor microenvironment and fibroblasts (Fb) contribute to tumor promotion in thyroid cancer. However, the role of tumor-stroma derived EVs still needs to be deciphered. We hypothesized that the interaction of thyroid tumor cells with Fb would liberate EVs with a specific proteomic profile, which would have an impact on EV-functionality in thyroid tumor progression-related events. Tumor (TPC-1, 8505c) and non-tumor (NThyOri) thyroid cells were co-cultured with human Fb. EVs, obtained by ultracentrifugation of conditioned media, were characterized by nanoparticle tracking analysis and western blotting. EV-proteomic analysis was performed by mass-spectrometry, and metalloproteinases (MMPs) were studied by zymography. EV-exchange was evaluated using immunofluorescence, confocal microscopy and FACS. EVs expressed classical exosome markers, with EVs from thyroid tumor cell-Fb co-cultures showing a proteomic profile related to extracellular matrix (ECM) remodeling. Bidirectional crosstalk between Fb and TPC-1 cells produced significantly more EVs than their isolated cells, and potentiated EV-functionality. In line with this, Fb-TPC-1 derived EVs induced MMP2 activation in NThyOri supernatants, and MMP2 activity could be evidenced in Fb and TPC-1 contact-independent co-cultures. Besides, MMP2 interactors allowed us to discriminate between EVs from thyroid tumoral and non-tumoral milieus. Interestingly, Fb internalized more EVs from TPC-1 than from NThyOri producing cells. Fb and thyroid tumor cell crosstalk produces specialized EVs with an ECM remodeling proteomic profile, enabling activation of MMP2 and possibly facilitating ECM-degradation, which is potentially linked with thyroid tumor progression.
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•Thyroid tumor cell-fibroblasts interplay releases specialized extracellular vesicles.•Extracellular vesicles acquire an extracellular matrix degradation profile.•Extracellular vesicles from thyroid tumor milieu promote metalloproteinase-2 activity.