Surrogate markers for brain damage

急性神经退行性疾病（如创伤性脑损伤、中风和心脏骤停引起的脑损伤）的诊断、预后和治疗评价将大大有助于脑损伤的微创替代措施，但目前缺乏这些措施。我的实验室设计了一种神经生物学方法，用于识别可在体液中测量的急性神经退行性变的新替代标志物。我们开始了第一次大规模的分析蛋白质释放从退化培养的大鼠皮层神经元，并使用现有的抗体收集沿着与二维聚丙烯酰胺凝胶电泳和质谱法检测60多个多肽释放优先从退化的神经元。通过基于抗体检测大鼠实验性创伤性脑损伤或脑缺血后脑脊液或血清中释放的多肽，以及人体循环停止的胸腹手术，已经建立了用于鉴定体内脑损伤替代标记物的方法的潜力。拟议的研究将扩展这些发现的临床前和临床应用。具体目标1将确定神经元细胞骨架蛋白和它们的蛋白水解片段，这些蛋白水解片段优先通过退化的培养神经元释放。具体目标2将定义由坏死和凋亡神经变性触发的蛋白质释放的蛋白质组。将开发新的抗体和夹心免疫测定法，用于对这两个目标产生的潜在标记物进行灵敏和特异性定量。具体目标3将通过对来自创伤性脑损伤实验模型的脑脊液、血清和尿液样本进行免疫测定来评价标志物，并建立标志物水平的时间和治疗依赖性变化与脑组织病理学之间的关系。具体目标4将开发人类生物标志物的免疫测定法，并在人类头部损伤的研究中对其进行检查。该项目的总体目标是确定新的替代标志物急性脑损伤可测量的体液，发展免疫测定，使其定量，并开始，以评估其临床前和临床实用程序的诊断，药理学，和替代措施的神经保护剂的疗效。

The diagnosis, prognosis, and therapeutic evaluation of acute neurodegenerative conditions such as traumatic brain injury, stroke, and cardiac arrest-induced brain damage would be aided considerably by minimally invasive surrogate measures for brain damage, but currently these are lacking. My laboratory has devised a neurobiological approach for identifying new surrogate markers for acute neurodegeneration measurable in accessible body fluids. We initiated the first large-scale analysis of protein release from degenerating cultured rat cortical neurons, and used an existing antibody collection along with methods of 2D PAGE and mass spectrometry to detect over 60 polypeptides released preferentially from degenerating neurons. The potential of the approach for identifying surrogate markers for brain injury in vivo has been established by antibody-based detection of released polypeptides in cerebrospinal fluid or serum following experimental traumatic brain injury or cerebral ischemia in rats, as well as thoraco-abdominal surgery with circulation arrest in humans. The proposed research would extend the preclinical and clinical applications of these findings. Specific Aim 1 will identify neuronal cytoskeletal proteins and their proteolytic fragments that are released preferentially by degenerating cultured neurons. Specific Aim 2 will define the proteomes for protein release triggered by necrotic and apoptotic neurodegeneration. Novel antibodies and sandwich immunoassays will be developed for the sensitive and specific quantitation of potential markers emerging from these two aims. Specific Aim 3 will evaluate the markers by immunoassays of cerebrospinal fluid, serum, and urine samples derived from an experimental model for traumatic brain injury, and establish relationships between time-and treatment-dependent changes in marker levels and brain histopathologies. Specific Aim 4 will develop immunoassays for human biomarkers and examine them in a study of human head injury. The overall goal of the project is to identify novel surrogate markers for acute brain damage measurable in accessible body fluids, develop immunoassays enabling their quantitation, and begin to assess their preclinical and clinical utilities as diagnostics, prognostics, and surrogate measures of neuroprotectant efficacy.