Human platelets aggregate at sites of blood vessel damage in response to a rise in their cytosolic calcium concentration.Controlling these cytosolic calcium rises would provide a method to inhibit platelet activation ...Human platelets aggregate at sites of blood vessel damage in response to a rise in their cytosolic calcium concentration.Controlling these cytosolic calcium rises would provide a method to inhibit platelet activation and prevent the unwanted blood clots that causes heart attack and strokes.Previously we have predicted that calcium accumulation within the lumen of an infolded portion of the platelet plasma membrane called the open canalicular system(OCS)is essential for maintaining this cytosolic calcium rise.Due to its nanometer dimensions of the OCS,it has been difficult to measure or interfere with the predicted luminal calcium accumulation.Here we utilise iron oxide magnetic nanoparticles coated with the known calcium chelator,citrate,to create calcium-binding nanoparticles.These were used to assess whether an OCS calcium store plays a role in controlling the dynamics of human platelet activation and aggregation.We demonstrate that citrate-coated nanoparticles are rapidly and selectively uptaken into the OCS of activated human platelets,where they act to buffer the accumulation of calcium there.Treatment with these calcium-binding nanoparticles reduced thrombin-evoked cytosolic calcium rises,and slowed platelet aggregation and clot retraction in human platelets.In contrast,nanoparticles that cannot bind calcium have no effect.This study demonstrates that the OCS acts as a key source of calcium for maintaining cytosolic calcium rises and accelerating platelet aggregation,and that calcium-binding nanoparticles targeted to the OCS could provide an anti-platelet therapy to treat patients at risk of suffering heart attacks or strokes.展开更多
基金Authors are deeply acknowledged to the Wellcome Trust for funding support(Seed Award in Science,Project grant no 207617/Z/17/Z).
文摘Human platelets aggregate at sites of blood vessel damage in response to a rise in their cytosolic calcium concentration.Controlling these cytosolic calcium rises would provide a method to inhibit platelet activation and prevent the unwanted blood clots that causes heart attack and strokes.Previously we have predicted that calcium accumulation within the lumen of an infolded portion of the platelet plasma membrane called the open canalicular system(OCS)is essential for maintaining this cytosolic calcium rise.Due to its nanometer dimensions of the OCS,it has been difficult to measure or interfere with the predicted luminal calcium accumulation.Here we utilise iron oxide magnetic nanoparticles coated with the known calcium chelator,citrate,to create calcium-binding nanoparticles.These were used to assess whether an OCS calcium store plays a role in controlling the dynamics of human platelet activation and aggregation.We demonstrate that citrate-coated nanoparticles are rapidly and selectively uptaken into the OCS of activated human platelets,where they act to buffer the accumulation of calcium there.Treatment with these calcium-binding nanoparticles reduced thrombin-evoked cytosolic calcium rises,and slowed platelet aggregation and clot retraction in human platelets.In contrast,nanoparticles that cannot bind calcium have no effect.This study demonstrates that the OCS acts as a key source of calcium for maintaining cytosolic calcium rises and accelerating platelet aggregation,and that calcium-binding nanoparticles targeted to the OCS could provide an anti-platelet therapy to treat patients at risk of suffering heart attacks or strokes.
