Volume 7 Issue 1
Lowering Low-Density Lipoprotein Particles in Plasma Using Dextran Sulphate Co-Precipitates Procoagulant Extracellular Vesicles
Jiong-Wei Wang, Ya-Nan Zhang, Siu Kwan Sze, Sander M. Van de Weg, Flora Vernooij, Arjan H. Schoneveld, Sock-Hwee Tan, Henri H. Versteeg, Leo Timmers, Carolyn S. P. Lam and Dominique P. V. De Kleijn
1Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, 119228 Singapore, Singapore
2Cardiovascular Research Institute, National University Heart Centre Singapore, 117599 Singapore, Singapore
3Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, 117593 Singapore, Singapore
4School of Biological Sciences, Nanyang Technological University, 637551 Singapore, Singapore
5Experimental Cardiology Laboratory, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
6Department of Medicine, National University of Singapore, 117599 Singapore, Singapore
7Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Centre, 2333 ZA Leiden, The Netherlands
8Department of Cardiology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands
9National Heart Centre Singapore, Duke-NUS Graduate Medical School, 169857 Singapore, Singapore
10Department of Cardiology, University Medical Center Groningen, 9713 GZ Groningen, The Netherlands
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*Authors to whom correspondence should be addressed.
Abstract
Plasma extracellular vesicles (EVs) are lipid membrane vesicles involved in several biological processes including coagulation. Both coagulation and lipid metabolism are strongly associated with cardiovascular events. Lowering very-low- and low-density lipoprotein ((V)LDL) particles via dextran sulphate LDL apheresis also removes coagulation proteins. It remains unknown, however, how coagulation proteins are removed in apheresis. We hypothesize that plasma EVs that contain high levels of coagulation proteins are concomitantly removed with (V)LDL particles by dextran sulphate apheresis. For this, we precipitated (V)LDL particles from human plasma with dextran sulphate and analyzed the abundance of coagulation proteins and EVs in the precipitate. Coagulation pathway proteins, as demonstrated by proteomics and a bead-based immunoassay, were over-represented in the (V)LDL precipitate. In this precipitate, both bilayer EVs and monolayer (V)LDL particles were observed by electron microscopy. Separation of EVs from (V)LDL particles using density gradient centrifugation revealed that almost all coagulation proteins were present in the EVs and not in the (V)LDL particles. These EVs also showed a strong procoagulant activity. Our study suggests that dextran sulphate used in LDL apheresis may remove procoagulant EVs concomitantly with (V)LDL particles, leading to a loss of coagulation proteins from the blood.
Keywords:extracellular vesicles; lipoprotein particles; low-density lipoprotein; LDL; LDL apheresis; coagulation; fibrinolysis