Effects of Piezo1 stimulation on platelet function and bioenergetics

Background: Platelets circulate in a haemodynamically complex environment subject to dynamic shear forces and respond to mechanical cues. Excess shear forces can lead to shear-induced platelet aggregation. Piezo1 has been described in platelets as inducing shear-induced Ca2+ fluxes. However, its interactions with GPCR and GPVI signalling pathways remain incompletely characterised, and the effect of Piezo1-induced Ca2+ flux on metabolic rewiring are unclear.
Methods: Washed human platelets from healthy donors were used and treated with the Piezo1 agonists Yoda1 (10 µM) and KC289 (10 µM), KC157 (10 µM), an inactive analogue and DMSO were used as controls. For aggregation and Ca2+ influx measurements, platelets were treated with Piezo1 agonists alone or in combination with a subthreshold dose of thrombin, convulxin/collagen or U46619; for bioenergetics, Piezo1 agonists alone were used.
Results: Piezo1 incubation potentiated subthreshold doses of GPCR signalling and showed an additive effect on Ca2+ flux. In bioenergetics, KC289 significantly increased ATP production from glycolysis versus DMSO. In the Glycolysis Stress Test, KC289 increased the acute ECAR response in glucose-free conditions and glycolysis. In the Mito Stress Test, KC289 showed a non-significant trend towards increased acute OCR. ROS assay showed a late rise in total ROS production at 12 h that was decreased in the presence of n-acetylcysteine.
Conclusion: Chemical Piezo1 agonism results in auxiliary Ca²⁺ entry that modestly increases ATP demand/supply, suggesting that mechanotransduction can modulate platelet bioenergetics via coordinated changes in glycolysis.