SCREENING FOR DRUGS IN A DROSOPHILA TRANSGENIC MODEL

在果蝇转基因模型中筛选药物

• 批准号: 6798018
• 负责人: GEORGE R JACKSON
• 金额: $ 19.17万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6798018
• 关键词: Alzheimer' s disease; Drosophilidae; antioxidants; bioassay; confocal scanning microscopy; cysteine endopeptidases; disease /disorder model; drug screening /evaluation; enzyme inhibitors; genetically modified animals; immunocytochemistry; microtubule associated protein; neural degeneration; neurofibrillary tangles; neuropathology; phosphorylation; protease inhibitor; scanning electron microscopy; transmission electron microscopy; western blottings

Mutations in the microtubule-associated protein tau occur in some cases of inherited frontotemporal dementia (FTD), demonstrating that tau abnormalities can cause neurodegeneration. Many FTD mutations occur in regulatory elements that alter splicing and thereby expression of tau isoforms, rather than tau coding sequence. One of the hallmark neuropathologic features of AIzheimer's disease (AD) is the neurofibrillary tangle (NFT), which contains hyperphosphorylated tau. Characterization of the events that modify tau-associated neurodegenerative processes is critical for understanding the pathophysiology of AD, as well as FTD and related diseases, such as progressive supranuclear palsy and corticobasal degeneration, and for the development of therapeutics. In order to test the hypothesis that neurodegeneration can be caused by aberrant expression of wild-type tau, the longest isoform of human tau was overexpressed in the fruit fly, producing degeneration of the eye and underlying brain, but failing to produce neurofibrillary tangles. However, tau phosphorylation by co-expression of shaggy, the Drosophila homologue of glycogen synthase kinase (GSK)-3beta, an important tau kinase in vitro, produced a more severely degenerated eye, as well as lesions resembling NFT (Jackson, G.R., et al. (2002): Human wild-type tau interacts with wingless pathway components and produces neurofibrillary pathology in Drosophila. Neuron 34: 509-519). Here, we propose to test selected kinase inhibitors, caspase inhibitors, and antioxidants for their ability to suppress the dual tau + shaggy bioassay phenotype. We also will test a library of commercially available, FDA-approved compounds (Microsource Discovery Systems) for their ability to suppress the external eye phenotype of tau + shaggy flies. Subsequently, we will determine the effects of identified compounds on cell death and conformation and solubility of tau. This project contributes to the commitment of the UCLA Alzheimer's Disease Research Center (ADRC) to foster research that will lead to identification of disease-modifying therapies for AD and related conditions. The Project will interact with Project 2 assessing similar compounds in transgenic mice.

微管相关蛋白tau在某些遗传性疾病中发生突变 额颞性痴呆(FTD)，表明tau异常可导致神经变性。许多FTD突变发生在调节元件中，这些元件改变了剪接从而改变了tau亚型的表达，而不是tau编码序列。AIzheimer病(AD)的一个标志性神经病理特征是神经纤维缠结(NFT)，其中含有过度磷酸化的tau。对改变tau相关神经退变过程的事件的表征对于了解AD、FTD和相关疾病(如进行性核上性瘫痪和皮质基底膜退行性变)的病理生理学以及治疗的发展至关重要。为了验证野生型tau基因异常表达可导致神经退行性变的假说，在果蝇中过表达了最长的人类tau亚型，从而导致眼睛和 潜在的大脑，但不能产生神经原纤维缠结。然而，通过共表达Shaggy，果蝇糖原合成酶激酶-3β的同源物，一种重要的tau激酶，在体外产生了更严重的退化眼睛，以及类似NFT的病变(Jackson，G.R.，等人)。(2002)：人类野生型tau与无翼途径组件相互作用，并在果蝇中产生神经原纤维病理。神经元34：509-519)。在这里，我们建议测试选定的激酶抑制剂、半胱氨酸氨基转移酶抑制剂和抗氧化剂抑制双tau+shaggy生物测定表型的能力。我们还将测试一个商业化的、FDA批准的化合物库(MicroSourceDiscovery Systems)，以了解它们抑制tau+毛茸茸苍蝇的外部眼部表型的能力。随后，我们将确定已确定的化合物对细胞死亡以及tau的构象和溶解性的影响。该项目有助于加州大学洛杉矶分校的承诺 阿尔茨海默病研究中心(ADRC)促进研究，将导致识别 治疗阿尔茨海默病及相关疾病的疾病修正疗法。该项目将与项目2互动 在转基因小鼠中评估类似的化合物。