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、预后和治疗评估将在很大程度上通过微创脑损伤替代措施得到帮助，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，用于识别急性神经变性的新替代标志物，可在可接触的体液中测量。我们首次对培养的退化大鼠皮质神经元的蛋白质释放进行了大规模分析，并使用现有的抗体收集以及2D PAGE和质谱方法检测了60多种优先从退化神经元中释放的多肽。通过对实验性创伤性脑损伤或脑缺血大鼠脑脊液或血清中释放的多肽进行基于抗体的检测，以及对人体进行循环停止的胸腹外科手术，已经建立了鉴定体内脑损伤替代标志物的方法的潜力。建议的研究将扩展这些发现的临床前和临床应用。特异性目的1将鉴定神经元细胞骨架蛋白及其蛋白水解片段，这些蛋白水解片段是由退化的培养神经元优先释放的。特异性Aim 2将定义坏死和凋亡性神经变性引发的蛋白质释放的蛋白质组。新的抗体和夹心免疫测定法将被开发出来，用于对这两个目标中出现的潜在标记物进行敏感和特异性的定量。特异性目标3将通过外伤性脑损伤实验模型中提取的脑脊液、血清和尿液样本的免疫分析来评估这些标志物，并建立标志物水平随时间和治疗的变化与脑组织病理学之间的关系。Specific Aim 4将开发人类生物标志物的免疫测定方法，并在人类头部损伤研究中对其进行检验。该项目的总体目标是确定可在体液中测量的急性脑损伤的新型替代标记物，开发使其定量的免疫测定方法，并开始评估其临床前和临床应用，作为诊断、预后和神经保护剂功效的替代测量。

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.