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

冷藏血小板调节血管稳定性的治疗潜力 创伤 创伤是全球1-44岁人群死亡的主要原因， 创伤性脑损伤是创伤后死亡的头号原因。血小板输注 平衡的血液产品比例被证明可以提高严重受伤患者的存活率 失血的创伤病人。在目前的美国血库实践中，血小板(Plt)储存在 在22°C的孵化器中孵化最多5天。在22℃下存储5天的Plts与存储相关联 病变，感染风险增加，止血功能全面下降。Plts的4°C储存 已经被提议作为22°C存储的等效替代方案，在某些情况下甚至更好。 在治疗方面，除了在止血方面发挥关键作用外，Plt还可以保护 血管内皮的完整性。血管不稳定是创伤的一个显著影响 损伤导致血管通透性、炎症、凝血功能障碍和终末器官 失败了。在颅脑损伤中，颅内出血(ICH)和脑水肿是导致 死亡率，这可能是可以通过输注Plt来解决的。我们之前的数据表明 4°C Plts调节血管稳定性和抑制内皮细胞(EC)通透性类似于 22°C Plts。这项建议旨在阐明血小板和冷藏的治疗效果。 (4oC)血小板对创伤性脑损伤血管稳定性的影响 可作为治疗颅脑损伤的一线治疗干预措施，以减少脑水肿，脑出血， 神经炎症和改善脑外伤患者的预后。目标1是设计的机械式目标 目的：研究4℃保存对Plts和Plts的结构、分子和细胞信号的影响。 血小板-内皮细胞相互作用和凝血的研究。相关Plt和EC信号转导的抑制剂 途径将被用来梳理哪些途径与Plt对内皮细胞的影响有关 渗透性。AIM 2旨在更深入地了解4°C Plts对 利用最先进的提睾肌活体成像技术检测血管内皮细胞 大脑，可视化4°C Plts对血管通透性、血栓形成和保存的影响 血管内皮细胞的糖基化反应。AIM 3旨在验证我们的主要假设，即4°C Plts 可用于调节颅脑损伤的血管稳定性，从而改善止血，减少 神经炎症和神经细胞丢失。在这份修订后的提案中，我们展示了我们的 完全有能力执行建议的研究，并已改进我们的实验以成功 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.