Reticulated platelets in coronary artery disease: a multidimensional approach unveils prothrombotic signalling and novel therapeutic targets

Kirmes K, Han J, Klug M, Bloxham CJ, Babyak O, Bernett J, Arend L, Manz Q, Raka L, Schwartz L, Hoffmann M, Rosenbaum M, Ruland J, Ciora OA, Louadi Z, Tsoy O, Newaz K, Modica J, Conca Dioguardi C, Peano C, Müller M, Santovito D, Viggiani G, Kühne S, Von Scheidt M, Nicolai L, Wu T, Baumbach J, Chiarito M, Laugwitz KL, Condorelli G, Raake PW, List M, Bernlochner I, Bongiovanni D (2025)

Publication Type: Journal article

Publication year: 2025

Journal

European Heart Journal Oxford University Press

Book Volume: 46

Pages Range: 4901-4917

Journal Issue: 45

DOI: 10.1093/eurheartj/ehaf694

Abstract

Background and Aims Reticulated platelets (RPs), hyperreactive and RNA-rich, are associated with increased risk of cardiovascular events and suboptimal response to antiplatelet therapy in coronary artery disease (CAD). However, the underlying mechanisms remain poorly defined. This study aimed to characterize the molecular and functional phenotype of RPs in CAD and assess their potential as therapeutic targets. Methods RPs and mature platelets (MPs) were isolated from CAD patients based on RNA content and CD41 expression. Paired RP vs MP comparisons were conducted within each donor. Transcriptomic profiling (RNA-seq) was integrated with high-dimensional proteomics (mass cytometry) and validated in independent cohorts. Functional studies, including flow cytometry-based platelet–platelet binding, in vitro thrombosis, platelet spreading, and intracellular phospho-protein profiling, assessed the impact of PI3K and GPVI pathways. Results were curated in Platlas, an open-access interactive web resource. Results Among 95 CAD patients, RPs exhibited elevated activation marker expression and enrichment of prothrombotic pathways compared with MPs. RNA-seq revealed upregulation of GP6, TBXA2R, and VWF transcripts, novel GPVI splicing, and RP-specific non-coding RNAs including novel circRNAs. Proteomic and functional assays confirmed heightened PI3K and GPVI signalling, with increased phosphorylation of AKT, PI3K, and SYK, and elevated reactive oxygen species production. RPs showed increased aggregation, spreading and greater recruitment in thrombus formation, which were significantly reduced by PI3K (LY294002) and GPVI (glenzocimab) inhibition. Conclusions This study provides the first mechanistic explanation for RP hyperreactivity, revealing a distinct molecular profile and identifying GPVI and PI3K inhibition as promising targets for tailored antiplatelet therapy in CAD patients with elevated RPs.

Authors with CRIS profile

Judith Bernett

Involved external institutions

Technische Universität München (TUM) DE Germany (DE) Klinikum der Universität München (LMU Klinikum) DE Germany (DE) Universität Hamburg (UHH) DE Germany (DE) Humanitas Research Hospital / IRCCS Istituto Clinico Humanitas IT Italy (IT) Klinikum rechts der Isar DE Germany (DE) Universität Augsburg DE Germany (DE) Institute of Genetic and Biomedical Research / Istituto di Ricerca Genetica e Biomedica (IRGB CNR) IT Italy (IT) Deutsches Zentrum für Herz-Kreislauf-Forschung (DZHK e.V.) DE Germany (DE)

How to cite

APA:

Kirmes, K., Han, J., Klug, M., Bloxham, C.J., Babyak, O., Bernett, J.,... Bongiovanni, D. (2025). Reticulated platelets in coronary artery disease: a multidimensional approach unveils prothrombotic signalling and novel therapeutic targets. European Heart Journal, 46(45), 4901-4917. https://doi.org/10.1093/eurheartj/ehaf694

MLA:

Kirmes, Kilian, et al. "Reticulated platelets in coronary artery disease: a multidimensional approach unveils prothrombotic signalling and novel therapeutic targets." European Heart Journal 46.45 (2025): 4901-4917.

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