Annexin A1-dependent tethering promotes extracellular vesicle aggregation revealed with single-extracellular vesicle analysis

Maximillian A Rogers, Fabrizio Buffolo, Florian Schlotter, Samantha K Atkins, Lang H Lee, Arda Halu, Mark C Blaser, Elena Tsolaki, Hideyuki Higashi, Kristin Luther, George Daaboul, Carlijn V C Bouten, Simon C Body, Sasha A Singh, Sergio Bertazzo, Peter Libby, Masanori Aikawa, Elena Aikawa (Corresponding author)

Research output: Contribution to journalArticleAcademicpeer-review

53 Citations (Scopus)


Extracellular vesicles (EVs) including plasma membrane-derived microvesicles and endosomal-derived exosomes aggregate by unknown mechanisms, forming microcalcifications that promote cardiovascular disease, the leading cause of death worldwide. Here, we show a framework for assessing cell-independent EV mechanisms in disease by suggesting that annexin A1 (ANXA1)-dependent tethering induces EV aggregation and microcalcification. We present single-EV microarray, a method to distinguish microvesicles from exosomes and assess heterogeneity at a single-EV level. Single-EV microarray and proteomics revealed increased ANXA1 primarily on aggregating and calcifying microvesicles. ANXA1 vesicle aggregation was suppressed by calcium chelation, altering pH, or ANXA1 neutralizing antibody. ANXA1 knockdown attenuated EV aggregation and microcalcification formation in human cardiovascular cells and acellular three-dimensional collagen hydrogels. Our findings explain why microcalcifications are more prone to form in vulnerable regions of plaque, regulating critical cardiovascular pathology, and likely extend to other EV-associated diseases, including autoimmune and neurodegenerative diseases and cancer.

Original languageEnglish
Article numbereabb1244
Number of pages15
JournalScience Advances
Issue number38
Publication statusPublished - 1 Sept 2020

Bibliographical note

Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).


FundersFunder number
Board of XBiotech Inc
Esperion Therapeutics
Kowa Pharmaceuticals
NanoView Biosciences
XBiotech Inc
National Institutes of Health62/715,428, 1S10RR029237-01, CNA35-OG488, R01 HL141917, R01 HL136431, R01 HL147095
Ionis Pharmaceuticals
Kowa Company


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