Interactions between neutrophil extracellular traps and activated platelets enhance procoagulant activity in acute stroke patients with ICA occlusion
Peng Zhou,
Tao Li,
Jiaqi Jin,
Yingmiao Liu,
Baorong Li,
Quanye Sun,
Jiawei Tian,
Hongtao Zhao,
Zhihui Liu,
Shuai Ma,
Shuoqi Zhang,
Valerie A Novakovic,
Jialan Shi,
Shaoshan Hu
Affiliations
Peng Zhou
Department of Neurosurgery, The Second Affiliated Hospital, Harbin Medical University, Harbin, PR China; The Key Laboratory of Myocardial Ischemia, Ministry of Education, Heilongjiang Province, Harbin, PR China
Tao Li
Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, PR China
Jiaqi Jin
Department of Neurosurgery, The Second Affiliated Hospital, Harbin Medical University, Harbin, PR China
Yingmiao Liu
Department of Stomatology, The First Affiliated Hospital, Harbin Medical University, Harbin, PR China
Baorong Li
Department of Stomatology, The First Affiliated Hospital, Harbin Medical University, Harbin, PR China
Quanye Sun
Department of Clinical Laboratory, Qingdao Municipal Hospital Group, Qingdao, PR China
Jiawei Tian
Department of Neurosurgery, The Second Affiliated Hospital, Harbin Medical University, Harbin, PR China
Hongtao Zhao
Department of Neurosurgery, The Second Affiliated Hospital, Harbin Medical University, Harbin, PR China
Zhihui Liu
Department of Neurosurgery, The Second Affiliated Hospital, Harbin Medical University, Harbin, PR China
Shuai Ma
Department of Neurosurgery, The Second Affiliated Hospital, Harbin Medical University, Harbin, PR China
Shuoqi Zhang
Department of Neurology, The Second Affiliated Hospital, Harbin Medical University, Harbin, PR China
Valerie A Novakovic
Department of Research, Brigham and Women's Hospital, VA Boston Healthcare System, Harvard Medical School, Boston, MA, USA
Jialan Shi
Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, PR China; Department of Surgery, Brigham and Women's Hospital, VA Boston Healthcare System, Harvard Medical School, Boston, MA, USA; Jialan Shi, MD, PhD., Department of Hematology, The First Affiliated Hospital, Harbin Medical University, 23 Youzheng Street, Nangang District, Harbin 150001, China; Department of Surgery, Brigham and Women's Hospital, VA Boston Healthcare System, Harvard Medical School, 1400 VFW Parkway, West Roxbury, MA 02132, USA.
Shaoshan Hu
Department of Neurosurgery, The Second Affiliated Hospital, Harbin Medical University, Harbin, PR China; Corresponding author: Shaoshan Hu, MD, PhD., Department of Neurosurgery, The Second Affiliated Hospital, Harbin Medical University, 246 Xuefu Road, Nangang District, Harbin 150001, China
Background: The role of neutrophil extracellular traps (NETs) in procoagulant activity (PCA) in stroke patients caused by thromboembolic occlusion of the internal carotid artery (ICA) remains unclear. Our objectives were to evaluate the critical role of NETs in the induction of hypercoagulability in stroke and to identify the functional significance of NETs during atherothrombosis. Methods: The levels of NETs, activated platelets (PLTs), and PLT-derived microparticles (PMPs) were detected in the plasma of 55 stroke patients and 35 healthy controls. NET formation and thrombi were analysed using immunofluorescence. Exposed phosphatidylserine (PS) was evaluated with flow cytometry and confocal microscopy. PCA was analysed using purified coagulation complex, thrombin, and fibrin formation assays. Findings: The plasma levels of NETs, activated PLTs, and PMP markers in the carotid lesion site (CLS) were significantly higher than those in the aortic blood. NETs were decorated with PS in thrombi and the CLS plasma of ICA occlusion patients. Notably, the complementary roles of CLS plasma and thrombin-activated PLTs were required for NET formation and subsequent PS exposure. PS-bearing NETs provided functional platforms for PMPs and coagulation factor deposition and thus increased thrombin and fibrin formation. DNase I and lactadherin markedly inhibited these effects. In addition, NETs were cytotoxic to endothelial cells, converting these cells to a procoagulant phenotype. Sivelestat, anti-MMP9 antibody, and activated protein C (APC) blocked this cytotoxicity by 25%, 39%, or 52%, respectively. Interpretation: NETs played a pivotal role in the hypercoagulability of stroke patients. Strategies that prevent NET formation may offer a potential therapeutic strategy for thromboembolism interventions. Funding: This study was supported by grants from the National Natural Science Foundation of China (61575058, 81873433 and 81670128) and Graduate Innovation Fund of Harbin Medical University (YJSKYCX2018-58HYD). Keywords: Neutrophil extracellular traps, Phosphotidylserine, Stroke, Hypercoagulable state, Thrombosis, Endothelial cells
