Confining thrombus morphospace through targeted inhibition of platelet mechanosensory signaling.

BACKGROUND

While current antiplatelets protect against thrombosis, their clinical utility is limited by an elevated risk of bleeding.

OBJECTIVES

To understand how structure-function relations in the hemostatic system may be leveraged into improve risk/benefit ratios of antiplatelet therapies.

METHODS

We developed a deep learning-based framework to track the activities of large numbers of platelets in vivo, enabling a detailed comparative assessment of the effects of therapeutic interventions on the evolving structural hierarchy of the hemostatic response.

RESULTS

Unlike conventional antiplatelets targeting paracrine signaling, selective pharmaceutical inhibition of platelet mechanosensory signaling via PI3KC2α preserved the initial build-up of thrombi following vascular injury to high-flow mesenteric veins. However, as this burst of hemostatic activity subsided, inhibition of platelet mechanosensory signaling caused localized reductions of platelet intracellular calcium ion levels ([Ca2+]i) in shear-exposed peripheral thrombus subregions, inhibiting the formation of platelet clusters capable of withstanding the drag forces of the blood flow. As a consequence, platelets in these subregions detached, became elongated, and/or slid along the thrombus surface. On a macrostructural level, this selective destabilization prevented sustained physical expansion of thrombi outside the perimeters of vascular injuries while preserving platelet packing density in thrombus subregions close to vascular injuries.

CONCLUSION

Collectively, our results highlight platelet mechanosensory signaling as a significant driver of sustained platelet population growth after the initial agonist-driven phase of thrombus expansion. We show that pharmaceutical targeting of this pathway enforced the convergence of thrombus growth trajectories toward a rheologically favorable setpoint without compromising the structural integrity of thrombus subregions that are critical for hemostasis.