Neuromuscular junction as a therapeutic target to improve post-traumatic outcomes

项目摘要 四肢损伤造成的严重出血是造成战场死亡和可预防的创伤的重要原因。 平民医疗中的死亡事件。止血带是止血最有效的方法。 院前环境中的出血和在骨科和血管外科造成的无血外科领域 手术；然而，止血带相关的缺血和随后的再灌注(IR)可导致严重的IR损伤。 这些IR损伤导致了止血带使用的限制。探索机制并找到有效的方法 治疗可以解决止血带使用的局限性，改善预后和术后生活质量 创伤性病人。神经肌肉接头的结构是为了传递来自运动神经的电信号。 烟碱型乙酰胆碱受体(NAChRs)的终末影响肌肉功能。我们的试点数据 在止血带诱导的6周IR小鼠中，证实了一些nAChR簇的碎裂。 因此，在目标1中，我们将确定nAChR簇的碎片与 长期使用止血带引起的IR患者的肌肉收缩功能障碍。此外，我们的初步研究已经 靶向一条特定的信号通路，将受损肌肉中nAChR簇的断裂联系在一起，即 炎性细胞因子-细胞周期蛋白依赖性激酶5(CDK5-catenin-Rapsyn)信号通路。在目标2中，我们将 检测炎性细胞因子-CDK5-连环蛋白-Rapsyn信号通路是否介导nAChR断裂 长期止血带诱导的IR中的簇状物。在目标3中，我们将研究一种新型抗肿瘤药物的治疗效果。 炎性药物通过抑制对神经肌肉接头功能和结构的长期恢复 促炎症细胞因子在止血带/IR损伤小鼠模型中的作用。我们将在体外和体内设计 针对抗炎将促进神经肌肉修复这一主要假设的研究 交叉口。总体而言，拟议中的研究将揭示长期致病的细胞和分子机制。 止血带诱导的IR中的神经肌肉连接障碍。这些研究将提供进一步的信息， 神经肌肉接头可能是止血带/IR损伤的潜在治疗靶点，特别是通过 一种新型抗炎药在这项建议中的应用。这种方法有很大的潜力来 解决临床止血带使用的局限性，从而改善预后和术后生活质量 创伤性病人。

Project Summary Severe hemorrhage from extremity injuries is a significant cause of battlefield deaths and preventable trauma fatalities in civilian medicine. Tourniquet use is the most effective means of arresting life-threatening limb hemorrhage in the pre-hospital setting and creating bloodless surgical fields in orthopedic and vascular surgeries; however, tourniquet-related ischemia and subsequent reperfusion (IR) can cause serious IR injuries. These IR injuries have led to the limitations of tourniquet use. Exploring the mechanisms and finding effective therapies can resolve the limitations of tourniquet use and improve outcomes and quality of life in post- traumatic patients. The neuromuscular junction is structured to transmit electrical signals from motor nerve terminals to nicotinic acetylcholine receptors (nAChRs) for affecting muscle function. Our pilot data demonstrated that some nAChR clusters are fragmented in mice with 6 weeks of tourniquet-induced IR. Therefore, in Aim 1, we will determine the relationship between the fragmentation of nAChR clusters and muscle contractile dysfunction in long-term tourniquet-induced IR. Additionally, our preliminary studies have targeted a specific signaling pathway that links fragmentation of nAChR clusters in the injured muscle, namely the inflammatory cytokine-cyclin-dependent kinase 5 (Cdk5-catenin-rapsyn signaling pathway. In Aim 2, we will test if the inflammatory cytokine-Cdk5-catenin-rapsyn signaling pathway mediates fragmentation of nAChR clusters in long-term tourniquet-induced IR. In Aim 3, we will investigate the therapeutic effect of a novel anti- inflammatory drug on long-term functional and structural recovery of the neuromuscular junction via inhibition of pro-inflammatory cytokines in mouse models of tourniquet/IR injury. We will design in vitro and in vivo studies to address our overarching hypothesis that anti-inflammation will promote repair of the neuromuscular junction. Overall, proposed studies will unveil cellular and molecular mechanisms responsible for long-term neuromuscular junction disorder in tourniquet-induced IR. These studies will provide further information that the neuromuscular junction could be a potential therapeutic target in tourniquet/IR injuries, especially through the application of a novel anti-inflammatory drug in this proposal. This approach has significant potential to resolve the limitations of clinical tourniquet use, thereby improving outcomes and quality of life in post- traumatic patients.