ALTERED TAU CONFORMATION AS AN EARLY MARKER IN AD

TAU 构象的改变作为 AD 的早期标志

• 批准号: 6098279
• 负责人: Lester Irvin Binder
• 金额: $ 15.71万
• 依托单位: RUSH UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6098279
• 关键词: Alzheimer's disease; aging; antigen antibody reaction; biomarker; brain disorder diagnosis; conformation; enzyme linked immunosorbent assay; epitope mapping; human tissue; immunocytochemistry; laboratory mouse; monoclonal antibody; neuroanatomy; paired helical filament; pathologic process; phosphorylation; postmortem; site directed mutagenesis; tau proteins

This project focuses on the formation of the fibrillar pathology in AD and on its chief component, tau protein. We hypothesize that progressive modification and polymerization of tau proteins into straight and paired helical filaments (PHFs) result from conformational changes that can be identified and quantified using specific monoclonal antibodies. Conformation-selective antibodies, most of which were produced in the laboratory of Peter Davies at Albert Einstein College of medicine, can be used to determine th evolution of cytoskeletal abnormalities as a function of AD progression in vulnerable brain regions, and to correlate these with structural features of tau. Using these antibodies, the stability of the PHF- or AD-selective tau conformations can be assessed and quantified through affinity measurements against recombinant monomeric tau, synthetic polymeric tau, and authentic PHF tau. We propose that early changes in tau phosphorylation and /or conformation lead to the formation of fibrillar pathology in AD, and that these changes are stabilized and can be observed very early in the disease process using early marker monoclonal antibodies prior to the onset of PHF formation. This hypothesis will be tested as follows; 1. We will model the structural features of tau in vitro that underlie binding of PHF-selective antibodies in situ. This will be accomplished through oligonucleotide-directed mutagenesis and affinity measurements. Both phosphorylation-dependent and conformation-dependent antibodies will be characterized. This information will suggest a progression for tau conformation-dependent antibodies will be characterized. This information will suggest a progression for tau conformational changes associated with the formation of the fibrillar pathology; 2. We propose to select novel monoclonal conformation-sensitive tau antibodies for analysis and use in Aims 1 and 3, respectively; 3. We will probe for early conformational changes in tau protein that occur during disease progression using quantitative immunohisto- and cytochemistry at the light and E.M. level; and, 4. We propose to confirm that the staining documented in situ is due to conformationally altered tau by using two site capture ELISAs in vulnerable brain regions in aged control populations, mild, moderate and severe AD. These studies will be performed in conjunction with projects to correlate the formation of the fibrillar pathology with galanin hyperinervation of the basal forebrain cholinergic neurons and expression of specific NGF receptors. Data will be further correlated with structural atrophy as measured by MRI and behavioral symptomatology as well as electrophysiological measures of function.

该项目的重点是AD中纤维病理的形成， 它的主要成分tau蛋白。 我们假设渐进式 将tau蛋白修饰和聚合成直链和成对的 螺旋丝（PHF）由构象变化引起， 使用特异性单克隆抗体进行鉴定和定量。 构象选择性抗体，其中大部分是在 爱因斯坦医学院彼得·戴维斯的实验室， 用于确定细胞骨架异常的演变， 在脆弱的大脑区域的AD进展，并将这些与 Tau的结构特征 使用这些抗体， 可以评估和定量PHF-或AD-选择性tau构象 通过对重组单体tau、合成的 多聚tau蛋白和真正的PHF tau蛋白。 我们认为tau蛋白的早期变化 磷酸化和/或构象导致原纤维的形成 这些变化是稳定的，可以观察到， 使用早期标记单克隆抗体， 在PHF形成之前。 这一假设将被检验为： 1. 我们将在体外模拟tau的结构特征， 是PHF选择性抗体原位结合的基础。 这将是 通过阿托伐他汀定向诱变和亲和 测量. 磷酸化依赖和构象依赖 将表征抗体。 这些信息将表明， tau构象依赖性抗体的进展将是 表征了 这些信息将提示tau蛋白的进展 与原纤维形成相关的构象变化 病理学; 2. 我们建议选择新的单克隆构象敏感 分别用于目的1和3中的分析和使用的tau抗体; 3. 我们 将探测tau蛋白的早期构象变化， 在疾病进展期间使用定量免疫组织化学和 光下和电镜下的细胞化学水平; 4。 我们建议确认 原位记录的染色是由于构象改变的tau蛋白 通过在老年人大脑脆弱区域使用两个位点捕获ELISA， 对照人群，轻度、中度和重度AD。 这些研究报告将 与项目结合进行，以将 基底前脑甘丙肽神经支配过度的纤维病理学 胆碱能神经元和特异性NGF受体的表达。 数据将 进一步与MRI测量的结构性萎缩相关， 行为行为学以及电生理学测量 功能