PDF icon Download PDF
Open Access
CC BY 4.0 · TH Open 2025; 09: a26616472
DOI: 10.1055/a-2661-6472
Original Article

Platelet RNA-Seq Reveals Genes Associated with Carotid Intima-Media Thickness: A Cross-Sectional Study

Zhanfei Tan   *
3   Wangjing Hospital, China Academy of Chinese Medical Sciences, Beijing, China
,
Fan Guo   *
1   Institute of Basic Medical Sciences of Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, China
2   Beijing Key Laboratory of Chinese Materia Pharmacology, Beijing, China
,
Jiaming Gao
1   Institute of Basic Medical Sciences of Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, China
2   Beijing Key Laboratory of Chinese Materia Pharmacology, Beijing, China
,
Lanlan Li
1   Institute of Basic Medical Sciences of Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, China
2   Beijing Key Laboratory of Chinese Materia Pharmacology, Beijing, China
,
Shujuan Xu
1   Institute of Basic Medical Sciences of Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, China
2   Beijing Key Laboratory of Chinese Materia Pharmacology, Beijing, China
,
Yehao Zhang
1   Institute of Basic Medical Sciences of Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, China
2   Beijing Key Laboratory of Chinese Materia Pharmacology, Beijing, China
,
Jianhua Fu
1   Institute of Basic Medical Sciences of Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, China
2   Beijing Key Laboratory of Chinese Materia Pharmacology, Beijing, China
,
Jianxun Liu
1   Institute of Basic Medical Sciences of Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing, China
2   Beijing Key Laboratory of Chinese Materia Pharmacology, Beijing, China
› Author Affiliations
Funding This research was funded by the National Natural Science Foundation of China (No. 82030124); China Academy of Chinese Medical Sciences (CACMS) Innovation Fund (No. CI2023C047YLL); Postdoctoral Fellowship Program of CPSF (No. GZC20242024).
› Further Information
Also available ateRef
  Permissions and Reprints
Preview

Abstract

Background

Although the association between platelet characteristics and the risk of developing atherosclerosis (AS) has been acknowledged, the specific role of platelets in AS development and progression remains unclear. Therefore, the aim of this study was to identify platelet characteristics in patients with and without AS to enhance the understanding of their pathophysiological functions and discover more sensitive biomarkers for AS diagnosis.

Methods

We conducted a cross-sectional study involving AS patients and healthy controls (N). Based on the Chinese guidelines for diagnosing carotid and vertebral artery AS and the 2010 American College of Cardiology Foundation/American Heart Association (ACCF/AHA) guidelines, we defined AS using carotid ultrasound to measure intima-media thickness (IMT). General information, including sex, age, height, and weight, was collected upon enrollment. A series of examinations, including physical exams, serum lipid profiles, blood glucose tests, liver and kidney function tests, platelet aggregation assays, and carotid artery ultrasounds, was performed. Platelets were extracted from plasma for RNA-seq analysis.

Results

No statistically significant differences in age, sex, body mass index, or blood pressure were observed between the groups. Total triglyceride, total cholesterol, low-density lipoprotein cholesterol, apolipoprotein B, red blood cell count, hemoglobin concentration, cholesterol levels, and carotid IMT were significantly greater, and vascular endothelial function was significantly lower in the AS group than in the N group. Using RNA-seq, we identified 784 differentially expressed genes—141 downregulated and 643 upregulated—with Gene Ontology enrichment showing significant associations with blood coagulation pathways, among others. Weighted correlation network analysis revealed four hub genes related to IMT: Integrin Subunit Alpha 2b (ITGA2B), Transforming Growth Factor Beta 1 (TGFB1), Platelet Factor 4 (PF4), and Glycoprotein IX Platelet (GP9).

Conclusion

Our findings indicate moderate correlations of elevated ITGA2B (r = 0.327, p = 0.004), TGFB1 (r = 0.362, p = 0.001), PF4 (r = 0.240, p = 0.038), and GP9 (r = 0.302, p = 0.008) levels with increased IMT, suggesting that these genes may serve as predictive biomarkers for AS.

Keywords

atherosclerosis - PF4 - platelet - RNA sequencing - WGCNA

Authors' Contributions

F.G. and Z.T. performed the experiments, analyzed the data, and wrote and revised the manuscript. J.G. and L.L. helped with the construction of the logical structure of the thesis and revision. Y.Z., J.F., and J.L. guided the design of the study, revised the manuscript, and provided financial support.


* These authors contributed equally to this article.


Supplementary Material



Publication History

Received: 02 December 2024

Accepted: 06 May 2025

Accepted Manuscript online:
22 July 2025

Article published online:
07 August 2025

© 2025. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)

Georg Thieme Verlag KG
Oswald-Hesse-Straße 50, 70469 Stuttgart, Germany

Bibliographical Record
Zhanfei Tan, Fan Guo, Jiaming Gao, Lanlan Li, Shujuan Xu, Yehao Zhang, Jianhua Fu, Jianxun Liu. Platelet RNA-Seq Reveals Genes Associated with Carotid Intima-Media Thickness: A Cross-Sectional Study. TH Open 2025; 09: a26616472.
DOI: 10.1055/a-2661-6472
 

  • References

  • 1 Libby P. The changing landscape of atherosclerosis. Nature 2021; 592 (7855) 524-533
    Search in Google Scholar
  • 2 Falk E. Pathogenesis of atherosclerosis. J Am Coll Cardiol 2006; 47 (8 Suppl): C7-C12
    Search in Google Scholar
  • 3 Zhu Y, Xian X, Wang Z. et al. Research progress on the relationship between atherosclerosis and inflammation. Biomolecules 2018; 8 (03) 80
    Search in Google Scholar
  • 4 Badimon L, Vilahur G. Thrombosis formation on atherosclerotic lesions and plaque rupture. J Intern Med 2014; 276 (06) 618-632
    Search in Google Scholar
  • 5 Gawaz M, Geisler T, Borst O. Current concepts and novel targets for antiplatelet therapy. Nat Rev Cardiol 2023; 20 (09) 583-599
    Search in Google Scholar
  • 6 Lu H, Daugherty A. Atherosclerosis. Arterioscler Thromb Vasc Biol 2015; 35 (03) 485-491
    Search in Google Scholar
  • 7 Wang N, Tall AR. Cholesterol in platelet biogenesis and activation. Blood 2016; 127 (16) 1949-1953
    Search in Google Scholar
  • 8 Li N. Platelets as an inter-player between hyperlipidaemia and atherosclerosis. J Intern Med 2024; 296 (01) 39-52
    Search in Google Scholar
  • 9 Gerdes N, Seijkens T, Lievens D. et al. Platelet CD40 exacerbates atherosclerosis by transcellular activation of endothelial cells and leukocytes. Arterioscler Thromb Vasc Biol 2016; 36 (03) 482-490
    Search in Google Scholar
  • 10 Marx C, Novotny J, Salbeck D. et al. Eosinophil-platelet interactions promote atherosclerosis and stabilize thrombosis with eosinophil extracellular traps. Blood 2019; 134 (21) 1859-1872
    Search in Google Scholar
  • 11 Siegel-Axel D, Daub K, Seizer P, Lindemann S, Gawaz M. Platelet lipoprotein interplay: trigger of foam cell formation and driver of atherosclerosis. Cardiovasc Res 2008; 78 (01) 8-17
    Search in Google Scholar
  • 12 Sun S, Zou X, Wang D. et al. IRGM/Irgm1 deficiency inhibits neutrophil-platelet interactions and thrombosis in experimental atherosclerosis and arterial injury. Biomed Pharmacother 2023; 158: 114152
    Search in Google Scholar
  • 13 Di Costanzo A, Indolfi C, Sorrentino S, Esposito G, Spaccarotella CAM. The effects of statins, ezetimibe, PCSK9-inhibitors, inclisiran, and icosapent ethyl on platelet function. Int J Mol Sci 2023; 24 (14) 11739
    Search in Google Scholar
  • 14 Wang L, Tang C. Targeting platelet in atherosclerosis plaque formation: current knowledge and future perspectives. Int J Mol Sci 2020; 21 (24) 9760
    Search in Google Scholar
  • 15 Martinez Bravo G, Annarapu G, Carmona E. et al. Platelets in thrombosis and atherosclerosis: A double-edged sword. Am J Pathol 2024; 194 (09) 1608-1621
    Search in Google Scholar
  • 16 Nardin M, Verdoia M, Cao D. et al. Platelets and the atherosclerotic process: An overview of new markers of platelet activation and reactivity, and their implications in primary and secondary prevention. J Clin Med 2023; 12 (18) 6074
    Search in Google Scholar
  • 17 Song P, Fang Z, Wang H. et al. Global and regional prevalence, burden, and risk factors for carotid atherosclerosis: a systematic review, meta-analysis, and modelling study. Lancet Glob Health 2020; 8 (05) e721-e729
    Search in Google Scholar
  • 18 Greenland P, Alpert JS, Beller GA. et al.; American College of Cardiology Foundation, American Heart Association. 2010 ACCF/AHA Guideline for Assessment of Cardiovascular Risk in Asymptomatic Adults: Executive Summary: A Report of the American College of Cardiology Foundation/American Heart Association Task Force on Practice Guidelines Developed in Collaboration With the American Society of Echocardiography, American Society of Nuclear Cardiology, Society of Atherosclerosis Imaging and Prevention, Society for Cardiovascular Angiography and Interventions, Society of Cardiovascular Computed Tomography, and Society for Cardiovascular Magnetic Resonance. J Am Coll Cardiol 2010; 56 (25) e50-e103
    Search in Google Scholar
  • 19 Koenen RR, Binder CJ. Platelets and coagulation factors: Established and novel roles in atherosclerosis and atherothrombosis. Atherosclerosis 2020; 307: 78-79
    Search in Google Scholar
  • 20 Chan NC, Weitz JI. Antithrombotic agents. Circ Res 2019; 124 (03) 426-436
    Search in Google Scholar
  • 21 Bhatia K, Ladd LM, Carr KH. et al. Contemporary antiplatelet and anticoagulant therapies for secondary stroke prevention: A narrative review of current literature and guidelines. Curr Neurol Neurosci Rep 2023; 23 (05) 235-262
    Search in Google Scholar
  • 22 Bonaca MP, Bauersachs RM, Anand SS. et al. Rivaroxaban in peripheral artery disease after revascularization. N Engl J Med 2020; 382 (21) 1994-2004
    Search in Google Scholar
  • 23 Chiarito M, Sanz-Sánchez J, Cannata F. et al. Monotherapy with a P2Y12 inhibitor or aspirin for secondary prevention in patients with established atherosclerosis: a systematic review and meta-analysis. Lancet 2020; 395 (10235): 1487-1495
    Search in Google Scholar
  • 24 Bai X, Zhang H, Li Z. et al. Platelet-derived extracellular vesicles encapsulate microRNA-34c-5p to ameliorate inflammatory response of coronary artery endothelial cells via PODXL-mediated P38 MAPK signaling pathway. Nutr Metab Cardiovasc Dis 2022; 32 (10) 2424-2438
    Search in Google Scholar
  • 25 Ma Q, Fan Q, Han X. et al. Platelet-derived extracellular vesicles to target plaque inflammation for effective anti-atherosclerotic therapy. J Control Release 2021; 329: 445-453
    Search in Google Scholar
  • 26 Wolf D, Ley K. Immunity and inflammation in atherosclerosis. Circ Res 2019; 124 (02) 315-327
    Search in Google Scholar
  • 27 Kong P, Cui ZY, Huang XF, Zhang DD, Guo RJ, Han M. Inflammation and atherosclerosis: signaling pathways and therapeutic intervention. Signal Transduct Target Ther 2022; 7 (01) 131
    Search in Google Scholar
  • 28 Breland UM, Michelsen AE, Skjelland M. et al. Raised MCP-4 levels in symptomatic carotid atherosclerosis: an inflammatory link between platelet and monocyte activation. Cardiovasc Res 2010; 86 (02) 265-273
    Search in Google Scholar
  • 29 Jin R, Xiao AY, Song Z. et al. Platelet CD40 mediates leukocyte recruitment and neointima formation after arterial denudation injury in atherosclerosis-prone mice. Am J Pathol 2018; 188 (01) 252-263
    Search in Google Scholar
  • 30 Ali A, Mounika N, Nath B. et al. Platelet-derived sTLT-1 is associated with platelet-mediated inflammation in coronary artery disease patients. Cytokine 2024; 178: 156581
    Search in Google Scholar
  • 31 He C, Medley SC, Hu T. et al. PDGFRβ signalling regulates local inflammation and synergizes with hypercholesterolaemia to promote atherosclerosis. Nat Commun 2015; 6: 7770
    Search in Google Scholar
  • 32 Li J, Tan M, Xiang Q, Zhou Z, Yan H. Thrombin-activated platelet-derived exosomes regulate endothelial cell expression of ICAM-1 via microRNA-223 during the thrombosis-inflammation response. Thromb Res 2017; 154: 96-105
    Search in Google Scholar
  • 33 Wang Z, Huang H. Platelet factor-4 (CXCL4/PF-4): an angiostatic chemokine for cancer therapy. Cancer Lett 2013; 331 (02) 147-153
    Search in Google Scholar
  • 34 Brandt E, Petersen F, Ludwig A, Ehlert JE, Bock L, Flad HD. The beta-thromboglobulins and platelet factor 4: blood platelet-derived CXC chemokines with divergent roles in early neutrophil regulation. J Leukoc Biol 2000; 67 (04) 471-478
    Search in Google Scholar
  • 35 Lecomte-Raclet L, Alemany M, Sequira-Le Grand A. et al. New insights into the negative regulation of hematopoiesis by chemokine platelet factor 4 and related peptides. Blood 1998; 91 (08) 2772-2780
    Search in Google Scholar
  • 36 Domschke G, Gleissner CA. CXCL4-induced macrophages in human atherosclerosis. Cytokine 2019; 122: 154141
    Search in Google Scholar
  • 37 Pitsilos S, Hunt J, Mohler ER. et al. Platelet factor 4 localization in carotid atherosclerotic plaques: correlation with clinical parameters. Thromb Haemost 2003; 90 (06) 1112-1120
    Search in Google Scholar
  • 38 Fox JM, Kausar F, Day A. et al. CXCL4/Platelet Factor 4 is an agonist of CCR1 and drives human monocyte migration. Sci Rep 2018; 8 (01) 9466
    Search in Google Scholar
  • 39 Gerdes N, Zhu L, Ersoy M. et al. Platelets regulate CD4+ T-cell differentiation via multiple chemokines in humans. Thromb Haemost 2011; 106 (02) 353-362
    Search in Google Scholar
  • 40 Sivapalaratnam S, Westbury SK, Stephens JC. et al.; NIHR BioResource. Rare variants in GP1BB are responsible for autosomal dominant macrothrombocytopenia. Blood 2017; 129 (04) 520-524
    Search in Google Scholar
  • 41 Quach ME, Li R. Structure-function of platelet glycoprotein Ib-IX. J Thromb Haemost 2020; 18 (12) 3131-3141
    Search in Google Scholar
  • 42 Huang L, Shao B. New insights of glycoprotein Ib-IX-V complex organization and glycoprotein Ibα in platelet biogenesis. Curr Opin Hematol 2024; 31 (06) 294-301
    Search in Google Scholar
  • 43 Gentilini F, Turba ME, Giancola F. et al. A large deletion in the GP9 gene in Cocker Spaniel dogs with Bernard-Soulier syndrome. PLoS ONE 2019; 14 (09) e0220625
    Search in Google Scholar
  • 44 Lin Q, Zhou R, Meng P. et al. Establishment of a Bernard-Soulier syndrome model in zebrafish. Haematologica 2022; 107 (07) 1655-1668
    Search in Google Scholar
  • 45 Ghalloussi D, Rousset-Rouvière C, Popovici C. et al. Bernard-Soulier syndrome: first human case due to a homozygous deletion of GP9 gene. Br J Haematol 2020; 188 (06) e87-e90
    Search in Google Scholar
  • 46 Burkard P, Schonhart C, Vögtle T. et al. A key role for platelet GPVI in neutrophil recruitment, migration, and NETosis in the early stages of acute lung injury. Blood 2023; 142 (17) 1463-1477
    Search in Google Scholar
  • 47 Morikawa M, Derynck R, Miyazono K. TGF-β and the TGF-β family: Context-dependent roles in cell and tissue physiology. Cold Spring Harb Perspect Biol 2016; 8 (05) a021873
    Search in Google Scholar
  • 48 Bobik A, Agrotis A, Kanellakis P. et al. Distinct patterns of transforming growth factor-beta isoform and receptor expression in human atherosclerotic lesions. Colocalization implicates TGF-beta in fibrofatty lesion development. Circulation 1999; 99 (22) 2883-2891
    Search in Google Scholar
  • 49 Kalinina N, Agrotis A, Antropova Y. et al. Smad expression in human atherosclerotic lesions: Evidence for impaired TGF-beta/Smad signaling in smooth muscle cells of fibrofatty lesions. Arterioscler Thromb Vasc Biol 2004; 24 (08) 1391-1396
    Search in Google Scholar
  • 50 Chen PY, Qin L, Li G. et al. Endothelial TGF-β signalling drives vascular inflammation and atherosclerosis. Nat Metab 2019; 1 (09) 912-926
    Search in Google Scholar
  • 51 Wang L, Wang H, Zhu M. et al. Platelet-derived TGF-β1 induces functional reprogramming of myeloid-derived suppressor cells in immune thrombocytopenia. Blood 2024; 144 (01) 99-112
    Search in Google Scholar
  • 52 Feng W, Chang C, Luo D. et al. Dissection of autophagy in human platelets. Autophagy 2014; 10 (04) 642-651
    Search in Google Scholar
  • 53 Lewis JC, Maldonado JE, Mann KG, Moertel CG. Ultrastructural cytochemistry of platelets and megakaryocytes in the carcinoid syndrome. Mayo Clin Proc 1976; 51 (09) 585-593
    Search in Google Scholar
  • 54 Qiao L, Ma J, Zhang Z. et al. Deficient chaperone-mediated autophagy promotes inflammation and atherosclerosis. Circ Res 2021; 129 (12) 1141-1157
    Search in Google Scholar
  • 55 Li X, Zhu X, Wei Y. Autophagy in atherosclerotic plaque cells: Targeting NLRP3 inflammasome for self-rescue. Biomolecules 2022; 13 (01) 15
    Search in Google Scholar
  • 56 Mahdi A, Wodaje T, Kövamees O. et al. The red blood cell as a mediator of endothelial dysfunction in patients with familial hypercholesterolemia and dyslipidemia. J Intern Med 2023; 293 (02) 228-245
    Search in Google Scholar
  • 57 Michel JB, Martin-Ventura JL. Red blood cells and hemoglobin in human atherosclerosis and related arterial diseases. Int J Mol Sci 2020; 21 (18) 6756
    Search in Google Scholar
  • 58 Kucharska-Newton AM, Monda KL, Campbell S. et al. Association of the platelet GPIIb/IIIa polymorphism with atherosclerotic plaque morphology: the Atherosclerosis Risk in Communities (ARIC) Study. Atherosclerosis 2011; 216 (01) 151-156
    Search in Google Scholar
  • 59 Blanchet X, Cesarek K, Brandt J. et al. Inflammatory role and prognostic value of platelet chemokines in acute coronary syndrome. Thromb Haemost 2014; 112 (06) 1277-1287
    Search in Google Scholar