Modeling Neurofibrillary Degeneration

神经原纤维变性建模

• 批准号: 6948775
• 负责人: SHU-HUI C YEN
• 金额: $ 27.75万
• 依托单位: MAYO CLINIC JACKSONVILLE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-15 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6948775
• 关键词: Alzheimer' s disease; aldehydes; calpain; disease /disorder model; enzyme linked immunosorbent assay; glioma; human tissue; immunoelectron microscopy; mass spectrometry; neural degeneration; neurofibrillary tangles; oxidative stress; phosphorylation; tau proteins; western blottings

DESCRIPTION (provided by applicant): Progressive supranuclear palsy shares its defining pathologic signature, neurofibrillary tangles (NFT) consisting primarily of hyperphosphorylated tau, with numerous neurological diseases, including Alzheimer's disease, corticobasal degeneration, Pick's disease as well as frontotemporal dementia and Parkinsonism linked to chromosome 17. To improve our understanding of the mechanism underlying NFT formation and its functional impacts we have developed cellular models that produce tau filaments with morphological and biochemical characteristics of human tauopathies. The models consist of conditional transfectants generated from human neuroglioma [H4] and neuronal [BE(2)-M17D] cells in which transgenic production of wild-type or mutant tau is regulated via the TetOff inducible mechanism. Preliminary studies demonstrated that treatment of these cells with 4-hydroxynonneal (HNE), proteosomal or calpain inhibitors enhances the assembly of disulfide-linked tau aggregates. The results suggest that cellular insults such as oxidative stress and deregulation of proteases may play a role in the formation of NFT. We will employ this cellular model to uncover the molecular mechanism underlying tau aggregation induced by various insults. The Specific Aims of our proposal are: (1). To test if factors implicated in the etiology and pathogenesis of human tauopathies exacerbate tau aggregation in conditional transfectants, (2). To determine if the enhanced aggregation is associated with changes in tau solubility/partition, phosphorylation, degradation and oligomerization, (3). To investigate whether such exacerbated tau aggregation is associated with altered level or state of activation/activity of particular kinases, proteases and/or proteasomes, and (4). To study if progression of the exacerbated assembly of tau aggregates can be blocked through deregulating kinases/proteases. The results are likely to provide valuable information for a rational design of therapeutics to treat neurofibrillary degeneration.

描述（由申请人提供）： 进行性的上可观的瘫痪分享了其定义的病理学特征，神经原纤维缠结（NFT），主要由多种磷酸化的tau组成，其中包括多种神经系统疾病，包括阿尔茨海默氏病，皮质性可质性疾病，皮质性疾病，选择的疾病以及额外的痴呆症和parkinsonism链接的机制。形成及其功能影响，我们开发了细胞模型，这些模型产生了人类陶氏病的形态和生化特征的tau丝。该模型由人类神经瘤[H4]和神经元[BE（2）-M17D]细胞产生的条件转染物组成，其中通过TETOFF诱导机制调节了野生型或突变体TAU的转基因产生。初步研究表明，这些细胞用4-羟基核（HNE），蛋白酶体或钙蛋白酶抑制剂对这些细胞进行处理，从而增强了二硫键连接的TAU聚集体的组装。结果表明，诸如氧化应激和蛋白酶消耗管制之类的细胞损伤可能在NFT的形成中起作用。我们将采用这种细胞模型来揭示由各种侮辱引起的TAU聚集的分子机制。我们提案的具体目的是：（1）。为了测试与条件转染剂中人类tau的病因学和发病机理有关的因素是否加剧了tau的聚集，（2）。为了确定增强的聚集是否与tau溶解度/分区，磷酸化，降解和低聚的变化有关（3）。调查这种加重的Tau聚集是否与特定激酶，蛋白酶和/或蛋白酶体的激活/活性的水平或活性的改变有关，以及（4）。为了研究是否可以通过放松调节激酶/蛋白酶来阻断Tau聚集体加剧的组装的进展。结果可能会提供有价值的信息，用于治疗神经原纤维变性的理性设计。