Extracellular vesicles induce fibroblasts metalloproteinases expression in thyroid tumour microenvironment

BRAVO MIANA RC; GUANTAY ML; GILARDONI MB; DE PAUL AL; BORIOLI GA; PELLIZAS CG; DONADIO AC

Barcelona

Congreso; International Society of Extracellular Vesicles Anual Meeting; 2018

International Society of Extracellular Vesicles

Background: The tumour microenvironment (TME), conformed by cellular and non-cellular components, promotes tumourigenesis in many solid malignancies. Extracellular vesicles (EVs) are related with intercellular communication in the TME. Fibroblasts (Fb), present in the TME, foster tumour growth stimulating cell proliferation and secreting extracellular matrix-remodelling proteases such as matrix metalloproteinases (MMPs). CD147, a glycoprotein implicated in MMPs expression, is related to thyroid tumour progression. Previous studies showed that thyroid tumour cell?Fb interactions promoted the secretion of MMPs toculture supernatants (CMs) and a migratory phenotype in tumour cells.Our goal was to identify EVs production and their role as mediators ofMMPs expression in thyroid TME.Methods:As an in vitro tumour?stroma cell interaction model, nontumour cells (NThyOri), thyroid papillary carcinoma cells (TPC-1) and thyroid anaplastic cells (8505c) were co-cultured with normal Fb. EVs,obtained by ultracentrifugation of cell and cell-Fb CMs, were characterized by transmission electron microscopy and dynamic light scattering.MMPs were studied by zymography in EVs and CMs of Fb incubated with EVs. CD147 expression was analysed in EVs by gold-immunocytochemistry and western blot.Results: EVs ranged between 50 and 600 nm. No MMP activity was detected in EVs, but Fb incubated with EVs secreted MMP9 and MMP2 to CMs. Interestingly, an increase in MMP9 and active MMP2 was observed in Fb treated with EVs collected from tumour cell?Fb cocultures. Equally, an increased MMP2 and MMP9 expression was previously detected by immunocytochemistry in Fb cocultured with thyroidtumour cells. CD147 expression was detected in EVs and significantly increased in TPC-1-Fb and 8505c-Fb cocultures, without changes in NThyOri-Fb cocultures.Summary/conclusion: The results suggest that EVs play an active role inintercellular communication events in thyroid TME, stimulating the Fb release of MMPs and the consequent tumour cell migration.Funding: Secretaría de Ciencia y Tecnología de la Universidad Nacionalde Córdoba (SECyT-UNC). Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Fundación Sales (Fundación para la Investigación en Cáncer).