A Novel Localization in Human Large Extracellular Vesicles for the EGF-CFC Founder Member CRIPTO and Its Biological and Therapeutic Implications

Simple Summary During tumorigenesis, communication among cells is fundamental. Tumor cells can communicate through the release of extracellular vesicles (EVs), carrying active mediators, that are able to move in the extracellular space and in the body fluids, thereby reaching cells different from the ones of origin. Tumor-derived EVs, upon interaction with tumor target cells, can profoundly change cell characteristics and behavior. Here, we isolated EVs released from human teratocarcinoma cells and we uncovered their ability to inhibit the cell migration of glioblastoma (GBM) cells. Teratocarcinoma EVs contain the oncofetal protein CRIPTO, that is involved in the observed reduction of GBM cell migration. Our results suggest a novel localization and function for CRIPTO in large EVs, and give precious hints for the development of novel therapeutic approaches, based on the control of tumor cell migration, to ultimately improve prognosis and quality of life of GBM patients. Tumor growth and metastasis strongly rely on cell-cell communication. One of the mechanisms by which tumor cells communicate involves the release and uptake of lipid membrane encapsulated particles full of bioactive molecules, called extracellular vesicles (EVs). EV exchange between cancer cells may induce phenotype changes in the recipient cells. Our work investigated the effect of EVs released by teratocarcinoma cells on glioblastoma (GBM) cells. EVs were isolated by differential centrifugation and analyzed through Western blot, nanoparticle tracking analysis, and electron microscopy. The effect of large EVs on GBM cells was tested through cell migration, proliferation, and drug-sensitivity assays, and resulted in a specific impairment in cell migration with no effects on proliferation and drug-sensitivity. Noticeably, we found the presence of the EGF-CFC founder member CRIPTO on both small and large EVs, in the latter case implicated in the EV-mediated negative regulation of GBM cell migration. Our data let us propose a novel route and function for CRIPTO during tumorigenesis, highlighting a complex scenario regulating its effect, and paving the way to novel strategies to control cell migration, to ultimately improve the prognosis and quality of life of GBM patients.