Extracellular Vesicles-Based

Biomarkers Represent a Promising

Liquid Biopsy in Endometrial

Cancer

Carolina Herrero Filgueira, PhD student, Translational Medical

Oncology Group (ONCOMET), Health Research Institute of Santiago de Compostela (IDIS)

Introduction

CTCs cfDNA EVs

EVs --> Heterogeneous group of cell-derived membranous structures comprising exosomes, microvesicles and apoptotic bodies

Mechanism for intercellular communication. Messengers in tumor progression and metastasis (pre-metastatic niche formation)

EVs as biomarkers

Introduction

Gustafson D. et al., Front Cardiovasc Med. 2017 Nov 9; 4: 71.

ISOLATION METHODS

Ultracentrifugation

Ultrafiltration

SEC

Precipitation-Based EVs

Isolation

Microfluidics-Based EVs Capture

Introduction

Pin Li et al.,Theronostic. 2017 Jan 26;7(3):789-804 Monguió-Tartajada M. et al., Cell Mol Lif Sci. 2019 Jun;76(12):2369-2382

Immunoaffinity-Based EVs Capture

Introduction

Introduction

Williams C., et al,. J Extracell Vesicles. 2018 Mar 7;7(1): 1442985

EVs Precipitated

5 min 4oC 15 min 16000xg 4oC

Methodology

100-500µl

High

COSTS

SPECIFICITY

SAMPLE

TIME

Economical

ExoGAG Competitors

20 min >1h

Low

>1ml

Specific equipment

Application of ExoGAG technology

- EC is the most common neoplasm of the female genital tract in the developed world

- Recurrent EC --> poor prognosis --> L1CAM and ANXA2 --> Need for a biomarker with high sensitivity and/or specificity

ENDOMETRIAL CANCER (EC)

Adapted from Siegel RL. et al., CA Cancer J Clin. 2018 Jul; 68(4): 284-296

Results

EVs isolated from Hec1A endometrial cancer cells

- Characterization - Ultracentrifugation vs. ExoGAG

Herrero et al. Cancers. 2019 Dec 12; 11 (12)

EVs isolated from plasma samples

Results

- Characterization - ExoGAG vs ExoQuick and ExoSpin

Herrero et al. Cancers. 2019 Dec 12; 11 (12)

ANXA2 levels are increased in circulating EVs of EC patients

Results

Herrero et al. Cancers. 2019 Dec 12; 11 (12)

ANXA2 levels in purified EVs from EC patients showed diagnostic value and correlated with tumor aggressiveness

Results

- Patients with non-endometrioid tumors and high risk of recurrence had higher ANXA2 levels in purified EVs

- Clinical value of EV-associated ANXA2

Herrero et al. Cancers. 2019 Dec 12; 11 (12)

BASELINE (SURGERY) N= 41 EC

N= 20 CTRL

Conclusions

ExoGAG is a robust and highly efficient technology for the isolation of EVs from different biological fluids such as conditioned culture medium and plasma.

ExoGAG technology demonstrated an optimal performance for the clinical implementation of EVs

purified from plasma of EC patients.

The specific expression of the EC biomarkers ANXA2 in the isolated EVs improved their specificity and sensibility as liquid biopsy.

High ANXA2 levels in plasmatic EVs were associated with non-endometrioid tumors and high risk of recurrence tumors, evidencing the potential use of EV-based ANXA2 expression as a diagnostic and prognostic liquid biopsy biomarker in EC.

Acknowledgments

Miguel Abal Posada Lorena Alonso Alconada

Alba Ferreirós López Carlos Casas Arozamena

Alexandre de la Fuente González

VHIR

Eva Colás Antonio Gil-Moreno

INA

Victor Sebastián Manuel Arruebo

MD Anderson Cancer Center

Gema Moreno-Bueno

Specials thanks to all patients who participated in

the study

CHUS Ana Vilar