Surrogate markers for brain damage

脑损伤的替代标志物

• 批准号: 6983296
• 负责人: ROBERT SIMAN
• 金额: $ 30.83万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6983296
• 关键词: biomarker; brain disorder diagnosis; brain injury; clinical chemistry; clinical research; cytoskeletal proteins; diagnosis design /evaluation; human subject; immunologic assay /test; laboratory rat; neural degeneration; patient oriented research; proteomics; trauma

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.

脑损伤的微创替代措施将大大有助于急性神经退行性疾病的诊断、预后和疗效评估，如创伤性脑损伤、中风和心脏骤停所致的脑损伤，但目前缺乏这些替代措施。我的实验室设计了一种神经生物学方法，用于识别可在可获得的体液中检测到的急性神经退行性变的新替代标记物。我们启动了对变性培养的大鼠皮质神经元蛋白质释放的第一次大规模分析，并使用现有的抗体收集以及2D PAGE和MS方法检测了60多个优先从变性神经元释放的多肽。通过对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进行基于抗体的检测，以及在人类胸腹手术中停止循环，建立了该方法在体内识别脑损伤替代标志物的可能性。拟议的研究将扩大这些发现的临床前和临床应用。特定目标1将识别神经元细胞骨架蛋白及其蛋白分解片段，这些蛋白分解片段优先由退化培养的神经元释放。特定目标2将定义由坏死性和凋亡性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分析方法，以敏感和特异地定量这两个目标产生的潜在标记物。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、血清和尿液样本的免疫分析来评估标记物，并建立标记物水平随时间和治疗而变化与脑组织病理学之间的关系。特定目标4将开发人类生物标记物的免疫分析方法，并在人类头部损伤的研究中进行检查。该项目的总体目标是确定可在可获得的体液中测量的急性脑损伤的新的替代标记物，开发能够对其进行定量的免疫分析方法，并开始评估其作为神经保护剂有效性的诊断、预后和替代措施的临床前和临床应用价值。