The Therapeutic Potential of Cold Stored Platelets in Regulating Vascular Instability in Trauma

冷保存血小板调节血管稳定性的治疗潜力 创伤 创伤是全世界1-44岁人群的主要死因， 创伤性脑损伤（TBI）是创伤后死亡的头号原因。血小板输注 平衡比例的血液制品已被证明可以提高严重受伤患者的存活率， 出血的外伤病人在目前的美国血库实践中，血小板（Plt）储存在 在22°C下孵育长达5天。Plts在22°C下储存5天与储存相关。 损伤、感染风险增加和止血功能总体下降。Plt的4°C储存 已被提议作为22 °C储存的等效替代品，在某些情况下是上级替代品。 在治疗上，除了在止血中的关键作用外，已知Plt还可保护患者的免疫功能。 血管内皮的完整性。血管不稳定是创伤性心脏病的标志性影响， 损伤导致血管通透性、炎症、凝血障碍和终末器官 失败在TBI中，颅内出血（ICH）和脑水肿是TBI的主要原因。 死亡率，这是潜在的解决Plt输血。我们之前的数据显示 4°C Plts调节血管稳定性并抑制内皮细胞（EC）渗透性， 22°C Plts.该建议旨在阐明血小板和冷藏血小板的治疗作用。 (4oC)血小板对创伤性脑损伤（TBI）血管稳定性的影响 可用作TBI的一线治疗干预，以减少脑水肿，ICH， 神经炎症和改善TBI患者的预后。目标1是一个机械目标， 研究4°C储存对Plts的结构、分子和细胞信号传导的影响， 血小板-内皮细胞相互作用和凝血。相关Plt和EC信号传导的抑制剂 将利用这些途径来梳理哪些途径与Plt对内皮细胞的作用相关， 磁导率目标2旨在更深入地了解4°C Plts对 通过使用提睾肌中的最新技术水平的活体成像来观察血管内皮， 脑，以观察4°C Plts对血管通透性、凝块形成和保存的影响 内皮糖萼的结构。目标3旨在验证我们的主要假设，即4°C Plts 可用于调节TBI中的血管稳定性，从而改善止血， 神经炎症和神经元细胞损失。在这份修订后的提案中，我们表明了我们 完全有能力执行拟议的研究，并完善了我们的实验， 回答提出的问题。

The Therapeutic Potential of Cold Stored Platelets in Regulating Vascular Stability in Trauma Trauma is the leading cause of death world-wide in individuals between the ages of 1-44, with traumatic brain injury (TBI) being the number one cause of death after trauma. Platelet transfusion and balanced ratios of blood products have been shown to increase survival in severely injured bleeding trauma patients. In the current US blood-banking practice, platelets (Plts) are stored in incubators at 22°C for up to 5 days. Storage of Plts at 22°C for 5 days is associated with a storage lesion, increased infectious risk, and an overall decline in hemostatic function. 4°C storage of Plts has been proposed as an equivalent and in some cases superior alternative to 22 °C storage. Therapeutically, in addition to their critical role in hemostasis, Plts are known to safeguard the integrity of the vascular endothelium. Vascular instability is a hallmark effect of traumatic injury leading to vascular permeability, inflammation, coagulation disturbances and end organ failure. In TBI, intracranial hemorrhage (ICH) and cerebral edema are the leading causes of mortality, which are potentially addressable by Plt transfusion. Our previous data demonstrate that 4°C Plts regulate vascular stability and inhibit endothelial cell (EC) permeability similar to 22°C Plts. This proposal will aim to elucidate the therapeutic effects of platelets and cold stored (4oC) platelets on vascular stability in traumatic brain injury (TBI) with the thought that platelets can be used as a first line therapeutic intervention in TBI to decrease cerebral edema, ICH, neuroinflammation and improve outcomes in TBI patients. Aim 1 is a mechanistic aim designed to investigate the structural, molecular and cellular signaling effects of 4°C storage on Plts and on Plt-endothelial cell interactions and coagulation. Inhibitors of relevant Plt and EC signaling pathways will be utilized to tease apart what pathways are relevant to Plt effects on endothelial permeability. Aim 2 is designed to provide a deeper understanding of the effects of 4°C Plts on the vascular endothelium by using state of the art intravital imaging in the cremaster muscle and brain, to visualize the effects of 4°C Plts on vascular permeability, clot formation and preservation of the endothelial glycocalyx. Aim 3 is designed to test out our primary hypothesis that 4°C Plts can be used to regulate vascular stability in TBI hence improving hemostasis, decreasing neuroinflammation and neuronal cell loss. In this revised proposal we have demonstrated our full capabilitiy to execute the proposed studies and have refined our experiments to successfully answer the questions posed.