Red blood cell Piezo1 Activation Drives Faster Coagulation and Structural Alterations in Blood Clots through Red Blood Cell Deformability Impairment

Red blood cells (RBCs) must be highly deformable to pass through capillaries narrower than their own diameter for delivering oxygen to the tissues. Ion channels are central regulators of RBC deformability and volume. Gain of function mutations in Piezo1, as seen in hereditary xerocytosis, cause RBC dehydration and have been associated with an increased risk of thromboembolism. Here we investigated how pharmacological RBC-Piezo1 activation affects RBC membrane potential, RBC ion content, RBC deformability, coagulation dynamics, and clot structure. Blood samples were collected from healthy donors. RBCs were isolated and incubated with Yoda1, a drug known to activate Piezo1. Membrane potential was measured using the Macey-Bennekou-Egée (MBE) method. Intracellular Na⁺/K⁺ content was determined by flame photometry. RBC deformability was measured by ektacytometry. Coagulation dynamics were investigated by rotational thromboelastometry (ROTEM), and clot structure was observed by electron microscopy. Yoda1 treatment led to RBC membrane hyperpolarization, reduced cell volume, a decrease in intracellular K⁺, an increase in Na⁺, and a reduction in RBC deformability. ROTEM analysis showed shorter clotting and lysis times in Yoda1-treated samples compared to samples treated with the vehicle (control condition), independently of RBC phosphatidylserine exposure. Electron microscopy revealed structural alterations in Yoda1-treated samples, with less compacted RBCs. Altogether, these findings indicate that Piezo1 activation disrupted RBC ion balance and membrane potential, leading to dehydration and decreased deformability. These alterations contributed to accelerated coagulation and compromised clot structure, suggesting a potential role of Piezo1 in modulating thrombotic risk in RBC-related disorders.