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Thromb Haemost 2023; 123(02): 207-218
DOI: 10.1055/a-1962-1613
Cellular Haemostasis and Platelets

Impaired Platelet Function and Thrombus Formation in PDE5A-Deficient Mice

Xiang Gui   #
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
,
Xiang Chu   #
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
,
Yuwei Du
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
,
Yuhan Wang
4   School of Medical Technology, Xuzhou Medical University, Xuzhou, People's Republic of China
,
Sixuan Zhang
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
,
Yangyang Ding
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
,
Huan Tong
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
,
Mengdi Xu
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
,
Yue Li
4   School of Medical Technology, Xuzhou Medical University, Xuzhou, People's Republic of China
,
Wen Ju
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
,
Zengtian Sun
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
,
Zhenyu Li
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
,
Lingyu Zeng   *
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
4   School of Medical Technology, Xuzhou Medical University, Xuzhou, People's Republic of China
,
Kailin Xu   *
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
,
Jianlin Qiao   *
1   Blood Diseases Institute, Xuzhou Medical University, Xuzhou, People's Republic of China
2   Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, People's Republic of China
3   Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, People's Republic of China
› Author Affiliations Funding This work was supported by the National Natural Science Foundation of China (grant no. 82170130, 81970124, 81400082, 81641151, and 81700178), the Natural Science Foundation of Jiangsu Province (grant no. BK20140219 and BK20170259), the funding for the Distinguished Professorship Program of Jiangsu Province, the Shuangchuang Project of Jiangsu Province, the Six Talent Peaks Project of Jiangsu Province (WSN-133), the 333 projects of Jiangsu Province (BRA2017542), the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (18KJA320010 and 17KJA320008), Jiangsu Province's Key Provincial Talents Program (ZDRCA2016054), Jiangsu Province's Graduate Scientific Research Innovation Program (KYCX22-2896 and KYCX21-2691), and Youth Science and Technology Innovation Team of Xuzhou Medical University.
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Abstract

Intracellular cyclic GMP (cGMP) inhibits platelet function. Platelet cGMP levels are controlled by phosphodiesterase 5A (PDE5A)-mediated degradation. However, the exact role of PDE5A in platelet function and thrombus formation remains poorly understood. In this study, we characterized the role of PDE5A in platelet activation and function. Platelets were isolated from wild type or PDE5A−/− mice to measure platelet aggregation, activation, phosphatidylserine exposure (annexin-V binding), reactive oxygen species (ROS) generation, platelet spreading as well as clot retraction. Cytosolic calcium mobilization was measured using Fluo-4 AM by a microplate reader. Western blot was used to measure the phosphorylation of VASP, ERK1/2, p38, JNK, and AKT. FeCl3-induced arterial thrombosis and venous thrombosis were assessed to evaluate the in vivo hemostatic function and thrombus formation. Additionally, in vitro thrombus formation was assessed in a microfluidic whole-blood perfusion assay. PDE5A-deficient mice presented significantly prolonged tail bleeding time and delayed arterial and venous thrombus formation. PDE5A deficiency significantly inhibited platelet aggregation, ATP release, P-selectin expression, and integrin aIIbb3 activation. In addition, an impaired spreading on collagen or fibrinogen and clot retraction was observed in PDE5A-deficient platelets. Moreover, PDE5A deficiency reduced phosphatidylserine exposure, calcium mobilization, ROS production, and increased intracellular cGMP level along with elevated VASP phosphorylation and reduced phosphorylation of ERK1/2, p38, JNK, and AKT. In conclusion, PDE5A modulates platelet activation and function and thrombus formation, indicating that therapeutically targeting it might be beneficial for the treatment of thrombotic diseases.

Keywords

PDE5A - cGMP - platelets - thrombosis - hemostasis

Author Contributions

X. Gui and X. Chu performed research, analyzed data, and wrote the manuscript. S. Zhang, Y. Wang, Y. Du, Y. Ding, H. Tong, M. Xu, Y. Li, W. Ju, Z. Sun, and Z. Li performed research and analyzed data, L. Zeng, K. Xu, and J. Qiao conceived and designed the study and wrote the manuscript.


# These authors share first authorship.


* These authors share senior authorship.


Supplementary Material



Publication History

Received: 29 June 2022

Accepted: 11 October 2022

Accepted Manuscript online:
17 October 2022

Article published online:
30 December 2022

© 2022. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

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