comprehensive-analysis-procoagulant-platelets-exhibiting-features

Comprehensive Analysis of Procoagulant Platelets Exhibiting Features of Necrosis, Apoptosis and Platelet Activation

Platelets circulating in the bloodstream are relatively quiescent but become "activated" upon encountering stimulants or "agonists" at the site of blood ...

Cleveland Clinic

Platelets are circulating cells central to haemostasis that follows vessel injury, as well as thrombosis that ensues as a consequence of pathological stasis or plaque rupture. Platelet responses to various stimuli that mediate these processes are all energy-intensive. Hence, platelets need to adapt their energy metabolism to fulfil the requirements of clot formation while overcoming the adversities of the thrombus niche such as restricted access to oxygen and nutrient. In the present review, we describe the changes in energy metabolism of platelets upon agonist challenge and their underlying molecular mechanisms. We briefly discuss the metabolic flexibility and dependency of stimulated platelets in terms of choice of energy substrates. Finally, we discuss how targeting the metabolic vulnerabilities of stimulated platelets such as aerobic glycolysis and/or beta oxidation of fatty acids could forestall platelet activation and thrombus formation. Thus, we present a case for modulating platelet energy metabolism using small-molecules as a novel anti-platelet strategy in the management of vaso-occlusive disorders like acute myocardial infarction, ischemic stroke, deep vein thrombosis and pulmonary embolism.

Energy metabolism in platelets fuels thrombus formation: Halting the thrombosis engine with small-molecule modulators of platelet metabolism.

Circulating platelets derived from bone marrow megakaryocytes play a central role in thrombosis and hemostasis. Despite being anucleate, platelets express several proteins known to have nuclear niche. These include transcription factors and steroid receptors whose non-genomic functions are being elucidated in platelets. Quite remarkably, components of some of the best-studied morphogen pathways, namely Notch, Sonic Hedgehog (Shh), and Wnt have also been described in recent years in platelets, which regulate platelet function in the context of thrombosis as well as influence their survival. Shh and Notch pathways in stimulated platelets establish feed-forward loops of autocrine/juxtacrine/paracrine non-canonical signaling that helps perpetuate thrombosis. On the other hand, non-canonical Wnt signaling is part of a negative feedback loop for restricting platelet activation and possibly limiting thrombus growth. The present review will provide an overview of these signaling pathways in general. We will then briefly discuss the non-genomic roles of transcription factors and steroid receptors in platelet activation. This will be followed by an elaborate description of morphogen signaling in platelets with a focus on their bearing on platelet activation leading to hemostasis and thrombosis as well as their potential for therapeutic targeting in thrombotic disorders.

Non-canonical non-genomic morphogen signaling in anucleate platelets: a critical determinant of prothrombotic function in circulation.

In-stent restenosis (ISR) and stent thrombosis (ST) are the most serious complications of coronary angioplasty and stenting. Although the evolution of drug-eluting stents (DES) has significantly restricted the incidence of ISR, they are associated with an enhanced risk of ST. In the present study, we explore the photothermal ablation of a thrombus using a nano-enhanced thermogenic stent (NETS) as a modality for revascularization following ST. The photothermal activity of NETS, fabricated by coating bare metal stents with gold nanorods generating a thin plasmonic film of gold, was found to be effective in rarefying clots formed within the stent lumen in various  assays including those under conditions mimicking blood flow. NETS implanted in the rat common carotid artery generated heat following exposure to a NIR-laser that led to effective restoration of blood flow within the occluded vessel in a model of ferric chloride-induced thrombosis. Our results present a proof-of-concept for a novel photothermal ablation approach by employing coated stents in the non-invasive management of ST.

Nanogold-coated stent facilitated non-invasive photothermal ablation of stent thrombosis and restoration of blood flow.

Platelets are essential component of circulation that plays a major role in hemostasis and thrombosis. During activation and its demise, platelets release platelet-derived microvesicles, with lysophosphatidylcholine (LPC) being a prominent component in their lipid composition. LPC, an oxidized low-density lipoprotein, is involved in cellular metabolism, but its higher level is implicated in pathologies like atherosclerosis, diabetes, and inflammatory disorders. Despite this, its impact on platelet function remains relatively unexplored. To address this, we studied LPC's effects on washed human platelets. A multimode plate reader was employed to measure reactive oxygen species and intracellular calcium using HDCF-DA and Fluo-4-AM, respectively. Flow cytometry was utilized to measure phosphatidylserine expression, mitochondrial membrane potential (ΔΨm), and mitochondrial permeability transition pore (mPTP) formation using FITC-Annexin V, JC-1, and CoCl/calcein-AM, respectively. Additionally, platelet morphology and its ultrastructure were observed via phase contrast and electron microscopy. Sonoclot and light transmission aggregometry were employed to examine fibrin formation and platelet aggregation, respectively. The findings demonstrate that LPC induced oxidative stress and increased intracellular calcium in platelets, resulting in increased phosphatidylserine expression and reduced ΔΨm. LPC triggered caspase-independent platelet death and mPTP opening via cytosolic and mitochondrial calcium, along with microvesiculation and reduced platelet counts. LPC increased the platelet's size, adopting a balloon-shaped morphology, causing membrane fragmentation and releasing its cellular contents, while inducing a pro-coagulant phenotype with increased fibrin formation and reduced integrin αIIbβ3 activation. Conclusively, this study reveals LPC-induced oxidative stress and calcium-mediated platelet death, necrotic in nature with pro-coagulant properties, potentially impacting inflammation and repair mechanisms during vascular injury.

Lysophosphatidylcholine induces oxidative stress and calcium-mediated cell death in human blood platelets.

Thrombosis, the formation of blood clots in arteries or veins, poses a significant health risk by disrupting the blood flow. It can potentially lead to major cardiovascular complications such as acute myocardial infarction or ischemic stroke (arterial thrombosis) and deep vein thrombosis or pulmonary embolism (venous thrombosis). Nevertheless, over the course of several decades, researchers have observed an association between different cardiovascular events and neurodegenerative diseases, which progressively harm and impair parts of the nervous system, particularly the brain. Furthermore, thrombotic complications have been identified in numerous clinical instances of neurodegenerative diseases, particularly Alzheimer's disease, Parkinson's disease, multiple sclerosis, and Huntington's disease. Substantial research indicates that endothelial dysfunction, vascular inflammation, coagulation abnormalities, and platelet hyperactivation are commonly observed in these conditions, collectively contributing to an increased risk of thrombosis. Thrombosis can, in turn, contribute to the onset, pathogenesis, and severity of these neurological disorders. Hence, this concise review comprehensively explores the correlation between cardiovascular diseases and neurodegenerative diseases, elucidating the cellular and molecular mechanisms of thrombosis in these neurodegenerative diseases. Additionally, a detailed discussion is provided on the commonly employed antithrombotic medications in the context of these neuronal diseases.

Role of Thrombosis in Neurodegenerative Diseases: An Intricate Mechanism of Neurovascular Complications.

Antibodies to β2-glycoprotein I (β2GPI) cause thrombosis in antiphospholipid syndrome; however, the role of β2GPI in coagulation in vivo is not understood. To address this issue, we developed β2GPI-deficient mice (Apoh-/-) by deleting exons 2 and 3 of Apoh using CRISPR/Cas9 and compared the development of thrombosis in wild-type (WT) and Apoh-/- mice using rose bengal- and FeCl3-induced carotid thrombosis, laser-induced cremaster arteriolar injury, and inferior vena cava (IVC) stasis models. We also compared tail bleeding times and activation of platelets from WT and Apoh-/- mice in the absence and presence of β2GPI. Apoh-/- mice demonstrated prolonged time to occlusion of the carotid after exposure to rose bengal or FeCl3 and reduced platelet and fibrin accumulation in cremasteric arterioles after laser injury. Significantly smaller thrombi formed in the IVC of Apoh-/- mice 48 hours after IVC occlusion. The activated partial thromboplastin time and prothrombin time, as well as activated partial thromboplastin time reagent and tissue factor-induced thrombin generation times, of Apoh-/- and WT plasma revealed no differences. However, we observed significant prolongation of tail bleeding in Apoh-/- mice and reduced P-selectin expression and fibrinogen binding to activated α2bβ3 on platelets from these mice after stimulation with thrombin; these changes were reversed by β2GPI. A protease activated receptor 3 (PAR3) antibody blocked thrombin-induced activation of WT platelets and the ability of β2GPI to restore thrombin-induced activation of Apoh-/- platelets. These studies demonstrate that β2GPI may promote platelet activation by enhancing the ability of PAR3 to present thrombin to PAR4.

Paresh P. Kulkarni

Department of Cardiovascular and Metabolic Sciences,
Lerner Research Institute,
Department of Cardiovascular and Metabolic Sciences, Lerner Research Institute

Cleveland Clinic

Comprehensive Analysis of Procoagulant Platelets Exhibiting Features of Necrosis, Apoptosis and Platelet Activation

Paresh P. Kulkarni

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Cardiovascular and Metabolic Sciences,Lerner Research Institute,Department of Cardiovascular and Metabolic Sciences, Lerner Research Institute

Cleveland Clinic

Comprehensive Analysis of Procoagulant Platelets Exhibiting Features of Necrosis, Apoptosis and Platelet Activation

Department of Cardiovascular and Metabolic Sciences,
Lerner Research Institute,
Department of Cardiovascular and Metabolic Sciences, Lerner Research Institute