Role and Function of Peroxisomal Ether Phospholipids in TBI

项目总结 创伤性脑损伤是人们关注的主要健康问题之一，在全世界每年有近170万人受到影响。 我们。到目前为止，还没有有效的治疗方法，主要是因为对分子的不完全理解。 损伤相关脑内生化变化的机制。在这项研究中，我们旨在确定 乙醚磷脂在脑外伤后神经退行性变中的作用和作用。乙醚磷脂起着至关重要的作用 在保持结构完整性和保护细胞膜免受氧化应激方面的作用。它们的合成是 由过氧化物酶体的酶机制启动。损伤引起的乙醚磷脂水平下降 在不同的神经退行性疾病中已经证明了过氧化体功能。然而，他们在 颅脑损伤的病理生理学尚未得到评估。在这项研究中，我们的目标是检验这一假设 过氧化物型醚磷脂损伤引起的乙醚磷脂丰度调节失调 合成参与了脑外伤后神经元的死亡。 我们将使用基于LC-MS/MS的脂组学方法来确定 脑创伤小鼠全脑皮质组织中乙醚磷脂的变化。我们还将确定 小鼠脑损伤后皮层过氧物组分中乙醚磷脂前体的研究 损伤皮质中过氧化物醚磷脂的合成功能。我们还将使用体外培养的神经元 牵张损伤模型研究乙醚磷脂丰度失调的机制及其作用 在损伤后神经元死亡中。在目标1中，我们将研究乙醚调节失调的机制 磷脂及其在小鼠脑损伤后神经元死亡中的作用。基于LC-MS/MS的联合脂组分析 将用生化方法检验过氧化物醚磷脂合成的假说 脑外伤后小鼠脑组织受损，导致乙醚磷脂丰度降低，从而导致脑损伤 受伤的大脑中的神经元死亡。在AIM 2中，我们将确定乙醚的神经保护功能 脑损伤小鼠磷脂前体的补充。我们假设乙醚磷脂的增加 丰富将减轻损伤相关的神经元死亡，并将恢复脑损伤的运动和认知功能 老鼠。我们将用乙醚磷脂前体-1-O-烷基甘油治疗脑外伤小鼠，并评估其神经保护作用 通过测量这些小鼠神经元死亡和功能恢复的程度来发挥作用。 我们的研究将首次确定乙醚磷脂在脑损伤中的作用和功能。我们相信 这项研究将为脑外伤相关神经元死亡提供新的机制见解，从而可能有助于我们 寻找治疗颅脑损伤的新靶点。

PROJECT SUMMARY Traumatic brain injury (TBI) is one of the major health concern affecting almost 1.7 million annually in the US. To date no effective therapy is available, mainly because of incomplete understanding of the molecular mechanisms of injury associated biochemical changes in the injured brain. In this study we aim to determine the role and function of ether phospholipids in neurodegeneration after TBI. Ether phospholipids play crucial role in maintaining structural integrity and protecting cellular membrane from oxidative stress. Their synthesis is initiated by the enzyme machinery of peroxisomes. Decreased level of ether phospholipids due to impaired peroxisomal functions have been demonstrated in different neurodegenerative diseases. However, their role in the pathophysiology of TBI has not been assessed. In this study our goal is to test the hypothesis that dysregulation of ether phospholipid abundance due to the impairment in peroxisomal ether phospholipid synthesis contributes to neuronal death after TBI. We will use liquid chromatography mass spectrometry (LC-MS/MS) based lipidomic approach to determine the changes in ether phospholipids in the whole cortical tissues of TBI mice. We will also determine the levels of ether phospholipid precursors in the peroxisomal fractions isolated from mouse cortices after TBI to assess peroxisomal ether phospholipid synthesizing function in the injured cortices. We will also use in vitro neuronal stretch injury model to investigate the mechanisms of dysregulation in ether phospholipid abundance and its role in neuronal death after injury. In AIM 1, we will investigate the mechanisms of dysregulation in ether phospholipid and its role in neuronal death after TBI in mice. LC-MS/MS based lipidomic analysis combined with biochemical methods will be used to test the hypothesis that peroxisomal ether phospholipid synthesis is impaired in the mouse brain after TBI causing decrease in ether phospholipid abundance that contributes to neuronal death in the injured brain. In AIM 2, we will determine the neuroprotective functions of ether phospholipid precursor supplementation in TBI mice. We hypothesize that increase in ether phospholipid abundance will attenuate injury associated neuronal death and will restore motor and cognitive functions in TBI mice. We will treat TBI mice with ether phospholipid precursor – 1-O-alkyl glycerol and assess its neuroprotective functions by measuring the extent of neuronal death and functional recovery in those mice. Our study will for the first time determine the role and function of ether phospholipids in TBI. We believe that this study will provide new mechanistic insight about TBI associated neuronal death and thus may help us to identify novel therapeutic target to treat TBI.