Cholangiocarcinoma (CCA) includes a heterogeneous group of biliary cancers with dismal prognosis. These tumors usually have a silent growth and are consequently commonly diagnosed in advanced phases, when the disease is already disseminated, limiting the accessibility to therapeutic options with curative-intent. Current non-invasive diagnostic tools show suboptimal accuracy for CCA detection, particularly at early tumor stages. Consequently, there is an urgent need to develop sensitive and specific non-invasive diagnostic tools for the detection of early CCAs, particularly in patients at high-risk such as primary sclerosing cholangitis (PSC). In this sense, extracellular vesicles (EVs), small membranous spheres found in biofluids which contain biuomolecules such as proteins, metabolites, nucleic acids and lipids from the cell of origin, have recently emerged as a potential source of biomarkers for human disorders. In accordance with this, in this dissertation, we have described nucleic acid and protein non-invasive biomarkers in serum and urine EVs for the prediction, early diagnosis and prognosis estimation of CCA, representing a novel tumor-cell derived liquid biopsy strategy.