TY - JOUR T1 - Conserved and Divergent Modulation of Calcification in Atherosclerosis and Aortic Valve Disease by Tissue Extracellular Vesicles JF - bioRxiv DO - 10.1101/2020.04.02.022525 SP - 2020.04.02.022525 AU - Mark C. Blaser AU - Fabrizio Buffolo AU - Arda Halu AU - Florian Schlotter AU - Hideyuki Higashi AU - Lorena Pantano AU - Louis A. Saddic AU - Samantha K. Atkins AU - Maximillian A. Rogers AU - Tan Pham AU - Eugenia Shvartz AU - Galina K Sukhova AU - Silvia Monticone AU - Giovanni Camussi AU - Simon C. Body AU - Jochen D. Muehlschlegel AU - Sasha A. Singh AU - Masanori Aikawa AU - Elena Aikawa Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/04/03/2020.04.02.022525.abstract N2 - Background Fewer than 50% of patients develop calcification of both atherosclerotic plaques and aortic valves, implying differential pathogenesis. While circulating extracellular vesicles (EVs) act as biomarkers of cardiovascular diseases, tissue-entrapped EVs associate with early mineralization, but their contents, function, and contributions to disease remain unknown.Results Global proteomics of human carotid artery endarterectomies and calcified aortic valves from a total of 27 donors/patients revealed significant over-representation of proteins with vesicle-associated pathways/ontologies common to both diseases. We exploited enzymatic digestion, serial (ultra)centrifugation and OptiPrep density-gradient separation to isolate EV populations from diseased arteries and valves. Mass spectrometry found 22 EV marker proteins to be highly enriched in the four least-dense OptiPrep fractions while extracellular matrix proteins predominated in denser fractions, as confirmed by CD63 immunogold electron microscopy and nanoparticle tracking analysis. Proteomics and miRNA-sequencing of OptiPrep-enriched tissue EVs quantified 1,104 proteins and 123 miR cargoes linked to 5,182 target genes. Pathway networks of proteins and miR targets common to artery and valve tissue EVs revealed a shared regulation of Rho GTPase and MAPK intracellular signaling cascades. 179 proteins and 5 miRs were significantly altered between artery and valve EVs; multi-omics integration determined that EVs differentially modulated cellular contraction and p53-mediated transcriptional regulation in diseased vascular vs. valvular tissue.Conclusions Our findings delineate a strategy to isolate, purify, and study protein and RNA cargoes from EVs entrapped in fibrocalcific tissues. Multi-omics and network approaches implicated tissue-resident EVs in human cardiovascular disease. ER -