Investigating the Potential and Pitfalls of EV-Encapsulated MicroRNAs as Circulating Biomarkers of Breast Cancer
Brian M. Moloney,
Katie E. Gilligan,
Doireann P. Joyce,
Clodagh P. O’Neill,
Killian P. O’Brien,
Sonja Khan,
Claire L. Glynn,
Ronan M. Waldron,
Ciarán M. Maguire,
Emma Holian,
Erin Naughton,
Mohamed Elhadi,
Andrea B. Grealish,
Carmel Malone,
Emma McDermott,
Peter Dockery,
Thomas Ritter,
Adriele Prina-Mello,
Michael J. Kerin,
Róisín M. Dwyer
Affiliations
Brian M. Moloney
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Katie E. Gilligan
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Doireann P. Joyce
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Clodagh P. O’Neill
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Killian P. O’Brien
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Sonja Khan
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Claire L. Glynn
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Ronan M. Waldron
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Ciarán M. Maguire
Laboratory for Biological Characterisation of Advanced Materials (LBCAM) and Advanced Materials and Bioengineering Research (AMBER) Centre, Trinity Translational Medicine Institute, Trinity College Dublin, James Street, Dublin D08 W9RT, Ireland
Emma Holian
School of Mathematics, Statistics and Applied Mathematics, National University of Ireland Galway, Galway H91 YR71, Ireland
Erin Naughton
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Mohamed Elhadi
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Andrea B. Grealish
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Carmel Malone
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Emma McDermott
Centre for Microscopy and Imaging, Discipline of Anatomy, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Peter Dockery
Centre for Microscopy and Imaging, Discipline of Anatomy, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Thomas Ritter
Regenerative Medicine Institute, School of Medicine, College of Medicine, Nursing and Health Sciences, National University of Ireland Galway, Galway H91 YR71, Ireland
Adriele Prina-Mello
Laboratory for Biological Characterisation of Advanced Materials (LBCAM) and Advanced Materials and Bioengineering Research (AMBER) Centre, Trinity Translational Medicine Institute, Trinity College Dublin, James Street, Dublin D08 W9RT, Ireland
Michael J. Kerin
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Róisín M. Dwyer
Discipline of Surgery, Lambe Institute for Translational Research, School of Medicine, National University of Ireland Galway, Galway H91 YR71, Ireland
Extracellular vesicles (EVs) shuttle microRNA (miRNA) throughout the circulation and are believed to represent a fingerprint of the releasing cell. We isolated and characterized serum EVs of breast tumour-bearing animals, breast cancer (BC) patients, and healthy controls. EVs were characterized using transmission electron microscopy (TEM), protein quantification, western blotting, and nanoparticle tracking analysis (NTA). Absolute quantitative (AQ)-PCR was employed to analyse EV-miR-451a expression. Isolated EVs had the appropriate morphology and size. Patient sera contained significantly more EVs than did healthy controls. In tumour-bearing animals, a correlation between serum EV number and tumour burden was observed. There was no significant relationship between EV protein yield and EV quantity determined by NTA, highlighting the requirement for direct quantification. Using AQ-PCR to relate miRNA copy number to EV yield, a significant increase in miRNA-451a copies/EV was detected in BC patient sera, suggesting potential as a novel biomarker of breast cancer.
