Extracellular Vesicles-Based Biomarkers Represent a Promising Liquid Biopsy in Endometrial Cancer.

Carolina Herrero,Carlos Casas-Arozamena,Alicia Abalo,Laura Muinelo-Romay,Martin Prieto,Antonio Gil-Moreno,Eva Colás,Alexandre De La Fuente,Miguel Abal,Juan Cueva,Manuel Arruebo,Gema Moreno-Bueno,Ana Vilar,Victor Sebastian

doi:10.3390/cancers11122000

Abstract

Tumor-derived extracellular vesicles (EVs) are secreted in large amounts into biological fluids of cancer patients. The analysis of EVs cargoes has been associated with patient´s outcome and response to therapy. However, current technologies for EVs isolation are tedious and low cost-efficient for routine clinical implementation. To explore the clinical value of circulating EVs analysis we attempted a proof-of-concept in endometrial cancer (EC) with ExoGAG, an easy to use and highly efficient new technology to enrich EVs. Technical performance was first evaluated using EVs secreted by Hec1A cells. Then, the clinical value of this strategy was questioned by analyzing the levels of two well-known tissue biomarkers in EC, L1 cell adhesion molecule (L1CAM) and Annexin A2 (ANXA2), in EVs purified from plasma in a cohort of 41 EC patients and 20 healthy controls. The results demonstrated the specific content of ANXA2 in the purified EVs fraction, with an accurate sensitivity and specificity for EC diagnosis. Importantly, high ANXA2 levels in circulating EVs were associated with high risk of recurrence and non-endometrioid histology suggesting a potential value as a prognostic biomarker in EC. These results also confirmed ExoGAG technology as a robust technique for the clinical implementation of circulating EVs analyses.

Highlights

Endometrial cancer (EC) is the most common neoplasm of the female genital tract in the developed world, and the incidence has risen over the past years
The efficiency of extracellular vesicles (EVs) purification by ExoGAG, expressed as the number of EVs per frame in the precipitate, was similar compared to the number of EVs in the pellet obtained by ultracentrifugation (Figure 1B)
Transmission electron microscopy (TEM) imaging showed that samples isolated with ExoGAG contained only EVs and crystallized organic remnants of ExoGAG agent (Figure 1D, right panels), while samples precipitated using ultracentrifugation showed a large amount of unwanted organic residues (Figure 1D, left panels)

Summary

Introduction

Endometrial cancer (EC) is the most common neoplasm of the female genital tract in the developed world, and the incidence has risen over the past years. Despite the progress in early detection and treatment, a significant number of cases of advanced ECs are still diagnosed. Mortality from this cancer has not improved in recent decades and is primarily driven by high-grade carcinomas that are more likely to present at an advanced stage and, are more likely to recur. The prognosis for recurrent EC is poor, especially for the 50% of these advanced ECs that present extra-pelvic disease recurrence To this regard, effort has been mainly dedicated to reach a consensus on EC risk classification to promote consistency for future clinical trials design, including their molecular characterization and its integration into clinicopathological profiling to develop prognostic and predictive biomarkers [1,2]. The presence of the L1 cell adhesion molecule (L1CAM) has been described as a strong independent predictor for distant recurrence and overall survival in EC, as well as a marker to select patients who could benefit from more extensive diagnostic and/or therapeutic procedures [3,4,5]

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