AB and ApoE in animal models of Alzheimer's disease

阿尔茨海默病动物模型中的 AB 和 ApoE

• 批准号: 6368893
• 负责人: ALEX E ROHER
• 金额: $ 33.74万
• 依托单位: BANNER SUN HEALTH RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6368893
• 关键词: Alzheimer's disease; aging; amyloid proteins; amyloidosis; apolipoprotein E; brain; disease /disorder model; genetically modified animals; genotype; high performance liquid chromatography; histochemistry /cytochemistry; immunologic assay /test; laboratory mouse; mass spectrometry; neuropathology; neurophysiology; pathologic process; phenotype; plasma; presenilin; protein metabolism; protein purification; protein sequence; protein structure function; transforming growth factors

Alzheimer disease (AD) is a dementia affecting over 20 million people worldwide. In the USA, AD represents the fourth greatest cause of death in the elderly. Genetic, chemical and morphological studies strongly suggest a seminal role for amyloid-beta (Abeta) peptides in the pathogenesis of AD. The successful creation of beta-amyloid precursor protein (betaAPP), presenilin 1 (PS1), apolipoprotein E (ApoE) and transforming growth factor-beta1 (TGF-beta1) transgenic (tg) mice has provided an opportunity to investigate the pathophysiology of Abeta amyloidogenesis. Tg mice are of paramount importance in the testing of AD therapeutic agents such as anti-Abeta aggregating compounds, secretase inhibitors and anti-Abeta vaccination. Despite the wide adoption of tg mice models in the search for effective AD treatments, a rigorous chemical characterization and quantification study of the Abeta produced by the genetically and phenotypically diverse tg mice strains has not been attempted. It is hypothesized that: 1) due to differences in lifespan, phylogenetic distance, functional organization and distress conditions of the brain significant quantitative and qualitative differences will be found in Abeta structure and metabolism between humans and mice; 2) failure in the clearance of Abeta from the brain to the blood in tg mice causes the progressive accumulation of both soluble and insoluble Abeta in the brain; 3) the partial loss of some of amino acid domains of ApoE, in an environment rich in proteolytic enzymes such as the AD brain, results in structural changes which would affect the affinity of ApoE for Abeta and the clearance of the complexes. The research plan will characterize the chemical structure and quantify Abeta peptides in the tg mice: APP23, 2576, PDAPP, PS1, TGF-beta1 and the double tg PS1/betaAPP and TGF-beta1/betaAPP. The ApoE associated to Abeta will be quantified and characterized in the tg mice brains. In addition, the effects of Abeta vaccination in the doubly PS1/betaAPP tg mice will be evaluated in terms of the soluble and insoluble Abeta40 and Abeta42 load. To achieve these goals the water-soluble and water-insoluble Abeta and ApoE peptides will be isolated from the tg mice by fast- and high- performance liquid column chromatographies from either brain homogenates or plasma. All purified molecules and derived enzymatic peptides will be chemically characterized by mass spectrometry, amino acid analysis and amino acid sequencing. Quantification of peptides will be performed by Western blot or by sensitive europium immunoassay. These significant characterizations and comparisons will provide important insights in evolutionary, physiological and pathological terms that will be useful for understanding AD and for the interpretation of mitigating treatments in tg mouse models.

阿尔茨海默氏病（AD）是一种痴呆症，全球影响超过2000万人。 在美国，广告代表了老年人中第四大死亡原因。 遗传，化学和形态学研究强烈表明淀粉样β（ABETA）肽在AD发病机理中的开创性作用。 成功地创建了β-淀粉样蛋白前体蛋白（BetaApp），Presenilin 1（PS1），载脂蛋白E（APOE）和转化生长因子-BETA1（TGF-BETA1）转基因（TG）小鼠提供了一个机会，提供了一个机会来研究Abeta淀粉样蛋白酶生理学的病理学。 TG小鼠在测试AD治疗剂（例如抗ABETA聚集化合物，泌尿酶抑制剂和抗ABETA疫苗接种）中至关重要。尽管TG小鼠模型在寻找有效的AD治疗时广泛采用，但尚未尝试对遗传和表型上不同的TG小鼠菌株产生的ABETA进行严格的化学表征和量化研究。假设：1）由于生命周期的差异，系统发育距离，功能组织以及大脑的困扰条件，在人类和小鼠之间的Abeta结构和代谢中会发现显着的定量和定性差异； 2）在TG小鼠中清除Abeta从大脑到血液的失败会导致大脑中可溶性和不溶性ABETA的逐步积累； 3）在富含蛋白水解酶（例如AD大脑）的环境中，APOE的某些氨基酸域的部分损失导致结构变化，这会影响ApoE对Abeta的亲和力和复合物的清除。 该研究计划将表征化学结构并量化TG小鼠中的ABETA肽：App23，2576，PDAPP，PS1，TGF-BETA1和Double TG PS1/betaapp和TGF-Beta1/betaapp。 与Abeta相关的APOE将在TG小鼠大脑中进行量化和表征。 此外，将根据可溶性和不溶性Abe​​ta40和Abeta42负载评估Abeta疫苗接种在双重PS1/BETAAPP TG小鼠中的影响。 为了实现这些目标，从脑匀浆或血浆中的快速和高性能液体色谱柱色谱分离出水溶性和不溶于水的Abeta和ApoE肽。 所有纯化的分子和衍生的酶肽将通过质谱，氨基酸分析和氨基酸测序化学特征。肽的定量将通过蛋白质印迹或敏感的欧洲免疫测定法进行。 这些重要的特征和比较将提供进化，生理和病理术语的重要见解，这些见解对于理解AD和解释TG小鼠模型中的缓解治疗方法非常有用。