TY - JOUR
T1 - Reticulated platelets in coronary artery disease
T2 - a multidimensional approach unveils prothrombotic signalling and novel therapeutic targets
AU - Kirmes, Kilian
AU - Han, Jiaying
AU - Klug, Melissa
AU - Bloxham, Conor J.
AU - Babyak, Olena
AU - Bernett, Judith
AU - Arend, Lis
AU - Manz, Quirin
AU - Raka, Leonora
AU - Schwartz, Leon
AU - Hoffmann, Markus
AU - Rosenbaum, Marc
AU - Ruland, Jürgen
AU - Ciora, Octavia Andreea
AU - Louadi, Zakaria
AU - Tsoy, Olga
AU - Newaz, Khalique
AU - Modica, Jessica
AU - Conca Dioguardi, Carola
AU - Peano, Clelia
AU - Müller, Michaela
AU - Santovito, Donato
AU - Viggiani, Giacomo
AU - Kühne, Stephanie
AU - Von Scheidt, Moritz
AU - Nicolai, Leo
AU - Wu, Tianjiao
AU - Baumbach, Jan
AU - Chiarito, Mauro
AU - Laugwitz, Karl Ludwig
AU - Condorelli, Gianluigi
AU - Raake, Philip W.J.
AU - List, Markus
AU - Bernlochner, Isabell
AU - Bongiovanni, Dario
N1 - Publisher Copyright:
© The Author(s) 2025. Published by Oxford University Press on behalf of the European Society of Cardiology. All rights reserved.
PY - 2025/12/1
Y1 - 2025/12/1
N2 - 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.
AB - 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.
KW - Coronary artery disease
KW - GPVI
KW - Mass cytometry
KW - Reticulated platelets
KW - Thrombocytes
KW - Transcriptomics
UR - https://www.scopus.com/pages/publications/105023532725
U2 - 10.1093/eurheartj/ehaf694
DO - 10.1093/eurheartj/ehaf694
M3 - Article
C2 - 40886063
AN - SCOPUS:105023532725
SN - 0195-668X
VL - 46
SP - 4901
EP - 4917
JO - European Heart Journal
JF - European Heart Journal
IS - 45
ER -