Establishment of Alzheimer model cell system and its application to therapeutics

阿尔茨海默病模型细胞系统的建立及其在治疗中的应用

基本信息

批准号：
07557332
负责人：
YOSHIKAWA Kazuaki
金额：
$ 0.96万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

Alzheimer's disease (AD) is characterized by the progressive dementia as a result of massive neuronal death in the brain. Clarification of the pathogenesis of this disease is one of the most urgent medical problems. Amyloid beta protein (Abeta) is the principal component of amyloid fibrils that are deposited in the brain affected by AD.Thus Abeta might be closely associated with the pathogenesis of the disease. Abeta is derived from the precursor, termed the amyloid precursor protein (APP). In an attempt to elucidate the pathological implications of intracellular accumulation of APP in postmitotic neurons, we transferred APP cDNA into rat hippocampal neurons under cultured conditions by using a replication-defective adenovirus vector. Neurons accumulating the membrane-bound form of APP showed greater responsiveness to exogenous glutamate than non-expressing control neurons, suggesting that elevated calcium levels potentially cause neurodegeneration. Moreover, we demonstrated that overexpression of APP by the same APP-expressing adenovirus vector induces death of human cultured neurons derived from human embryonal carcinoma cells within several days. When an APP-expressing adenovirus wazs injected into a rat hippocampus, some of the infected neurons in the hippocampal formation underwent severe degeneration displaying shrunk perikarya along with synaptic abnormalities in a few days. These experimental systems enable us to study detailed molecular mechanisms of APP-induced neurodegeneration, and will be useful for studies on pathogenesis of AD and development of therapeutics.

阿尔茨海默氏病（AD）的特征是由于大脑中大量神经元死亡而进行性痴呆。澄清该疾病的发病机理是最紧迫的医疗问题之一。淀粉样β蛋白（ABETA）是淀粉样蛋白原纤维的主要成分，这些淀粉样蛋白原纤维沉积在受AD影响的大脑中。因此，Abeta可能与疾病的发病机理密切相关。 Abeta源自前体，称为淀粉样蛋白前体蛋白（APP）。为了阐明APP在有丝分裂后神经元中APP的细胞内积累的病理意义，我们通过使用复制缺陷型腺病毒载体将APP cDNA转移到培养条件下的大鼠海马神经元中。与非表达对照神经元相比，累积膜结合APP的神经元对外源谷氨酸的反应性更大，这表明钙水平升高可能会导致神经变性。此外，我们证明了通过同一表达APP的腺病毒载体对APP的过表达会导致人类培养的神经元在几天内源自人类胚胎癌细胞的死亡。当表达应用的腺病毒WAZ注射到大鼠海马中时，一些海马形成中的一些感染神经元在几天内经历了严重的变性，表现出缩减的周围植核以及突触异常。这些实验系统使我们能够研究APP诱导的神经变性的详细分子机制，并将对AD和疗法发育的发病机理研究很有用。