Kinase Inhibitors against Neurodegeneration

激酶抑制剂对抗神经退行性变

• 批准号: 6736607
• 负责人: GARY S LYNCH
• 金额: $ 23.55万
• 依托单位: THURIS CORPORATION
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6736607
• 关键词: Niemann Pick disease; SDS polyacrylamide gel electrophoresis; cyclin dependent kinase; disease /disorder prevention /control; enzyme activity; glycogen synthase; immunocytochemistry; kinase inhibitor; laboratory mouse; mitogen activated protein kinase; neural degeneration; neurofibrillary tangles; pathologic process; phosphorylation; polymerase chain reaction; serine threonine protein kinase; tau proteins; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): Niemann-Pick type C (NPC) is a rare and fatal neurovisceral storage disorder that is currently untreatable, in most cases (95%), NPC is caused by mutations of the NPC1 gene, which encodes a glycoprotein playing an important role in cholesterol transport. NPC brains are also characterized by axonal abnormalities, neurofibrillary tangles (NFTs), and progressive neurodegeneration. The mechanisms leading to pathology in NPC are not well understood, although altered cholesterol metabolism in NPC mutant cells leads to axonal and neuronal degeneration as well as to hyperphosphorylation of the tau protein, which is the major component of NFTs. Interestingly, the NFTs observed in NPC and in Alzheimer's disease (AD) are chemically and morphologically indistinguishable and, in AD, they correlate with the severity of dementia. In brains from NPC1-/- mice, the activity of several protein kinases is altered and these alterations may result in tau modifications and neuronal death. In particular, the activities of cyclin-dependent kinase 5 (cdk5) and mitogen-activated protein kinase (MAPK) are elevated in NPCl-defective cells. This suggests that increased activity of selected protein kinases plays a critical role in NPC pathogenesis. The proposed experiments are directed at the following questions: (1) What are the contributions of cdk5, GSK3-beta, and MAP kinases to tau hyperphosphorylation and NFT formation? (2) What are the contributions of these kinases in the development of pathology taking place in NPC1-/- mice? Therefore, our specific aims are to (i) determine the effects of a variety of protein kinase inhibitors on a well established in vitro model of tau hyperphosphorylation and NFT formation using cultured hippocampal slices from ApoE-/- mice, (ii) test the effects of the most potent inhibitors of each class of protein kinases obtained in Aim 1 on a similar model that uses cultured hippocampal and cerebellar slices from NPC1-/- mice, and (iii) test the hypothesis that selected protein kinase inhibitors will prevent tau hyperphosphorylation, neuronal degeneration, microglial and astroglial reaction, and motor deficits in NPC1-/- mice in vivo. The proposed studies have the potential to (i) identify the pathway responsible for tau hyperphosphorylation and tangle formation, (ii) provide critical information concerning the role of specific protein kinases in other pathological manifestations of NPC disease, and (iii) open the way to broad-based drug discovery programs directed at essential aspects of NPC and AD.

描述（由申请人提供）：C型尼曼-匹克（NPC）是一种罕见的致命性神经内脏储存障碍，目前无法治疗，在大多数情况下（95%），NPC是由NPC 1基因突变引起的，NPC 1基因编码一种糖蛋白，在胆固醇转运中发挥重要作用。NPC脑的特征还在于轴突异常、神经纤维缠结（NFT）和进行性神经变性。虽然NPC突变细胞中胆固醇代谢的改变导致轴突和神经元变性以及tau蛋白的过度磷酸化，但导致NPC病理学的机制尚不清楚，tau蛋白是NFT的主要成分。 有趣的是，在NPC和阿尔茨海默病（AD）中观察到的NFT在化学和形态学上是不可区分的，并且在AD中，它们与痴呆的严重程度相关。在NPC 1-/-小鼠的大脑中，几种蛋白激酶的活性发生了改变，这些改变可能导致tau蛋白修饰和神经元死亡。特别是，细胞周期蛋白依赖性激酶5（cdk 5）和丝裂原活化蛋白激酶（MAPK）的活性在NPCl缺陷细胞中升高。这表明所选蛋白激酶的活性增加在NPC发病机制中起关键作用。本实验针对以下问题：（1）cdk 5、GSK 3-β和MAP激酶对tau蛋白过度磷酸化和NFT形成的贡献是什么？(2)这些激酶在NPC 1-/-小鼠病理学发展中的作用是什么？因此，我们的具体目标是（i）使用来自ApoE-/-小鼠的培养海马切片确定各种蛋白激酶抑制剂对tau过度磷酸化和NFT形成的良好建立的体外模型的作用，（ii）测试在目标1中获得的每类蛋白激酶的最有效抑制剂对使用来自NPC 1-/-的培养的海马和小脑切片的类似模型的作用。小鼠，和（iii）测试所选蛋白激酶抑制剂将在体内预防NPC 1-/-小鼠中的tau过度磷酸化、神经元变性、小胶质细胞和星形胶质细胞反应以及运动缺陷的假设。拟议的研究有可能（i）确定负责tau蛋白过度磷酸化和缠结形成的途径，（ii）提供有关特定蛋白激酶在NPC疾病其他病理表现中作用的关键信息，以及（iii）为针对NPC和AD基本方面的广泛药物发现计划开辟道路。