Isolation and Characterization of Cholinesterases From Plaques and Tangles

斑块和缠结中胆碱酯酶的分离和表征

基本信息

  • 批准号:
    7491670
  • 负责人:
  • 金额:
    $ 15.63万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2007
  • 资助国家:
    美国
  • 起止时间:
    2007-09-01 至 2010-08-31
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): It has been over two decades since it was demonstrated using histochemical methods that plaques and tangles, the pathological hallmarks of Alzheimer's disease (AD), contain activity specific to cholinergic enzymes acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). The same methodology demonstrated that these AD-related cholinesterases (AD-ChE) possess different properties when compared with cholinesterases found in normal neuronal cell bodies and axons, are likely to originate in glial cells and may possess unusual activities. Biochemical studies have indicated that ChEs may be involved in the pathology of AD, particularly as it relates to the genesis of plaques and production of their main constituent, the amyloid- (A) peptide. It has been suggested that in addition to current ChE inhibitor therapies aimed at ameliorating the cortical cholinergic deficit in AD, it would also be desirable and perhaps more beneficial to inhibit ChEs in plaques and tangles, due to their possible participation in the disease process. However, research regarding the ChEs in plaques and tangles is currently at a stand-still because the only methods to date that have allowed distinction between AD-ChEs and normal ChEs are based on histochemical demonstration of activity in tissue sections. There is no method available to specifically isolate ChEs from plaques and tangles. Therefore, there is no direct information on the biochemical properties of AD-ChEs and whether they make a specific contribution to disease pathology. Furthermore, the development and pre-clinical testing of ChE inhibitors that can specifically target AD-ChEs requires a method which can isolate these enzymes from tissue. Taking advantage of recent observations at our laboratory, the fact that some ChEs are anchored to tissues (such as muscle) with the aide of extracellular matrix proteins heparan sulphate proteoglycans (HSPG) and that HSPG are also present in plaques and tangles, the experiments in the present application will test two novel methods for isolation of ChEs from plaques, tangles and glia. The specific aims of the proposed research will test the following hypotheses: 1. Cholinesterases from plaques, tangles and glial cells but not neurons and axons in fixed and fresh frozen tissue sections will be passively released into storage buffer over time. 2. Cholinesterases from plaques, tangles and glial cells but not neurons and axons in fixed and fresh frozen tissue will be released upon treatment with various glycosaminoglycans, heparinases or proteases. 3. Released cholinesterases will display molecular and enzymatic characteristics and inhibition patterns identical to those demonstrated histochemically in plaques, tangles and glia and biochemically in AD brain. The successful isolation and characterization of AD-ChEs will allow mechanistic studies of the possible detrimental effects of these ChE species; will aide in design and testing of new inhibitors specifically targeting AD-ChEs; and, is likely to lead to investigation of AD-ChEs as biomarkers of AD in blood and CSF. Plaques and tangles of Alzheimer's disease contain the cholinergic enzymes acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) with altered properties that have been suggested to participate in the disease process. However, currently there are no methods available to isolate AChE and BuChE specifically from plaques and tangles, and the present application aims to fill in this gap. The successful isolation and characterization of AChE and BuChE from plaques and tangles will allow mechanistic studies of the possible detrimental effects of these enzyme species, will aide in design and testing of new inhibitors specifically targeting cholinesterases in plaques and tangles, and is likely to lead to investigation of plaque and tangle cholinesterases as biomarkers of AD in blood and cerebrospinal fluid.
描述(申请人提供):二十多年前,人们利用组织化学方法证明阿尔茨海默病(AD)的病理标志斑块和缠结含有胆碱能酶乙酰胆碱酯酶(AChE)和丁酰胆碱酯酶(BuChE)特异性活性。同样的方法表明,这些AD相关的胆碱酯酶(AD-ChE)具有不同的性质相比,在正常的神经元细胞体和轴突中发现的胆碱酯酶,可能起源于神经胶质细胞,并可能具有不寻常的活动。生化研究表明,胆碱酯酶可能参与AD的病理学,特别是因为它涉及斑块的发生和其主要成分淀粉样蛋白(A)肽的产生。已经表明,除了目前旨在改善AD中皮质胆碱能缺陷的ChE抑制剂疗法之外,还期望并且可能更有益的是抑制斑块和缠结中的ChE,因为它们可能参与疾病过程。然而,关于斑块和缠结中ChE的研究目前处于停滞状态,因为迄今为止唯一允许区分AD-ChE和正常ChE的方法是基于组织切片中活性的组织化学证明。没有方法可以从斑块和缠结中特异性分离ChE。因此,没有关于AD-ChE的生化特性以及它们是否对疾病病理学有特定贡献的直接信息。此外,可以特异性靶向AD-ChE的ChE抑制剂的开发和临床前测试需要可以从组织中分离这些酶的方法。利用我们实验室最近的观察结果,一些ChE在细胞外基质蛋白硫酸乙酰肝素蛋白聚糖(HSPG)的帮助下锚定到组织(如肌肉)上,并且HSPG也存在于斑块和缠结中,本申请中的实验将测试两种用于从斑块、缠结和神经胶质分离ChE的新方法。本研究的具体目的是验证以下假设:1.固定和新鲜冷冻组织切片中来自斑块、缠结和神经胶质细胞而非神经元和轴突的胆碱酯酶将随时间被动释放到储存缓冲液中。2.在固定和新鲜冷冻组织中,来自斑块、缠结和神经胶质细胞而非神经元和轴突的胆碱酯酶将在用各种糖胺聚糖、肝素酶或蛋白酶处理后释放。3.释放的胆碱酯酶将显示分子和酶的特性和抑制模式相同的斑块,缠结和神经胶质细胞和AD脑生化组织化学。AD-ChE的成功分离和表征将允许对这些ChE物质的可能有害作用进行机理研究;将有助于设计和测试特异性靶向AD-ChE的新抑制剂;并且可能导致将AD-ChE作为血液和CSF中AD的生物标志物进行研究。阿尔茨海默病的斑块和缠结含有胆碱能酶乙酰胆碱酯酶(AChE)和丁酰胆碱酯酶(BuChE),其具有已被建议参与疾病过程的改变的性质。然而,目前没有可用于从斑块和缠结中特异性分离AChE和BuChE的方法,本申请旨在填补这一空白。从斑块和缠结中成功分离和鉴定AChE和BuChE将允许对这些酶种类可能的有害作用进行机理研究,将有助于设计和测试特异性靶向斑块和缠结中胆碱酯酶的新抑制剂,并可能导致对斑块和缠结胆碱酯酶作为血液和脑脊液中AD生物标志物的研究。

项目成果

期刊论文数量(4)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Synergistic inhibition of butyrylcholinesterase by galantamine and citalopram.
Butyrylcholinesterase is associated with β-amyloid plaques in the transgenic APPSWE/PSEN1dE9 mouse model of Alzheimer disease.
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CHANGIZ GEULA其他文献

CHANGIZ GEULA的其他文献

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{{ truncateString('CHANGIZ GEULA', 18)}}的其他基金

Cognitive SuperAging: A model to explore resilience and resistance to aging and Alzheimers disease
认知超级老化:探索对衰老和阿尔茨海默病的恢复力和抵抗力的模型
  • 批准号:
    10901316
  • 财政年份:
    2023
  • 资助金额:
    $ 15.63万
  • 项目类别:
Study to Uncover Pathways to Exceptional Cognitive Resilience in Aging (SUPERAging)
研究揭示衰老过程中卓越认知弹性的途径(SUPERAging)
  • 批准号:
    10276525
  • 财政年份:
    2021
  • 资助金额:
    $ 15.63万
  • 项目类别:
Research Education Component
研究教育部分
  • 批准号:
    10469453
  • 财政年份:
    2021
  • 资助金额:
    $ 15.63万
  • 项目类别:
Biospecimen/Neuropathology Core
生物样本/神经病理学核心
  • 批准号:
    10276530
  • 财政年份:
    2021
  • 资助金额:
    $ 15.63万
  • 项目类别:
Biospecimen/Neuropathology Core
生物样本/神经病理学核心
  • 批准号:
    10687278
  • 财政年份:
    2021
  • 资助金额:
    $ 15.63万
  • 项目类别:
Research Education Component
研究教育部分
  • 批准号:
    10264374
  • 财政年份:
    2021
  • 资助金额:
    $ 15.63万
  • 项目类别:
Study to Uncover Pathways to Exceptional Cognitive Resilience in Aging (SUPERAging)
研究揭示衰老过程中卓越认知弹性的途径(SUPERAging)
  • 批准号:
    10687271
  • 财政年份:
    2021
  • 资助金额:
    $ 15.63万
  • 项目类别:
Research Education Component
研究教育部分
  • 批准号:
    10662493
  • 财政年份:
    2021
  • 资助金额:
    $ 15.63万
  • 项目类别:
Cognitive SuperAging: A model to explore resilience and resistance to aging and Alzheimers disease
认知超级老化:探索对衰老和阿尔茨海默病的恢复力和抵抗力的模型
  • 批准号:
    10359727
  • 财政年份:
    2020
  • 资助金额:
    $ 15.63万
  • 项目类别:
Characterized Adult Primary Human Microglia Cells for Research
用于研究的特征化成人原代人小胶质细胞
  • 批准号:
    10004183
  • 财政年份:
    2018
  • 资助金额:
    $ 15.63万
  • 项目类别:
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