Abstract Number: PB0776
Meeting: ISTH 2022 Congress
Theme: Hemostatic Systems in Cancer, Inflammation and Immunity » Platelets and Inflammation
Background: Endothelial cells (EC) suppress platelet activation and coagulation to prevent blood clotting. In inflammatory conditions, this negative regulation can be hampered and lead to thrombotic complications, but the mechanism is largely unknown.
Aims: Unravel the pathways by which (inflammatory) EC influence platelet signaling using vessel-on-a-chip and multi-omics approaches.
Methods: The transcriptomes of control-, TNFalpha- or LPS-treated Human Umbilical Venous EC (HUVEC) were characterized by RNA-sequencing. Platelet activation and thrombus formation on sub-confluent EC were monitored in whole-blood and high shear conditions. Changes in phosphorylation levels of platelets exposed to (treated) EC were assessed by TMT phosphoproteomic analysis. Platelet integrin activation (PAC1 mAb) in these conditions were measured by flow cytometry.
Results: Out of >18k robustly detectable genes, the HUVEC transcriptome showed ~0.7k and ~6.5k expression changes (LPS or TNF-treatment vs. Control respectively, adjusted p-value >0.01), in particular of inflammatory pathways and secretory mediators, after TNFalpha >> LPS treatment. Pathway analysis via ClusterProfiler and topGO revealed distinct inflammatory footprints of the treated EC. In high shear flow conditions on a collagen/tissue factor surface, the presence of sub-confluent EC potently suppressed platelet adhesion, thrombus and fibrin formation. When pre-treated with TNFalpha, the thrombo-protective and anticoagulant effect of the EC was significantly reverted. Flow cytometry revealed a ~80% reduction in agonist-induced integrin activation of platelets exposed to the control EC. This effect was half as strong after TNFalpha treatment, but not after LPS treatment. Key changes in the phosphoproteome (3.0k phospho-sites) in EC-exposed platelets are currently validated to uncover most relevant signaling alterations due to TNFalpha or LPS treatment of the cells.
Conclusion(s): The combined use of OMICs and microfluidic approaches reveals a high complexity of inflammation-modulated interactions between EC and platelets.
This project is funded from the EU Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement No 813409 (TICARDIO).
To cite this abstract in AMA style:
Schönichen C, Provenzale I, Ludt A, Solari F, Marini F, Cleuren A, Jurk K, Stoll M, Sickmann A, Kuijpers M, Heemskerk J. Using a multi-omics approach to understand endothelial dysregulation of platelet activation in inflammatory conditions [abstract]. https://abstracts.isth.org/abstract/using-a-multi-omics-approach-to-understand-endothelial-dysregulation-of-platelet-activation-in-inflammatory-conditions/. Accessed March 21, 2024.« Back to ISTH 2022 Congress
ISTH Congress Abstracts - https://abstracts.isth.org/abstract/using-a-multi-omics-approach-to-understand-endothelial-dysregulation-of-platelet-activation-in-inflammatory-conditions/