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.

在某些遗传性病例中，微管相关蛋白 tau 会发生突变。 额颞叶痴呆 (FTD)，证明 tau 蛋白异常可导致神经退行性变。许多 FTD 突变发生在调节元件中，这些元件改变剪接，从而改变 tau 异构体的表达，而不是改变 tau 编码序列。艾茨海默病 (AD) 的标志性神经病理学特征之一是神经原纤维缠结 (NFT)，其中含有过度磷酸化的 tau 蛋白。表征 tau 相关神经退行性过程的事件对于理解 AD 的病理生理学以及 FTD 和相关疾病（例如进行性核上性麻痹和皮质基底节变性）以及治疗方法的开发至关重要。为了验证神经变性可能是由野生型 tau 的异常表达引起的假设，人类 tau 的最长亚型在果蝇中过度表达，导致眼睛和眼睛的变性。 底层大脑，但无法产生神经原纤维缠结。然而，通过与糖原合酶激酶 (GSK)-3β（一种重要的体外 tau 激酶）果蝇同源物 shaggy 共表达而导致的 tau 磷酸化，产生了更严重的退化眼睛以及类似 NFT 的病变（Jackson，G.R. 等） al. (2002)：人类野生型 tau 蛋白与无翼通路成分相互作用，并在果蝇神经元中产生神经原纤维病理。 34：509-519）。在这里，我们建议测试选定的激酶抑制剂、半胱天冬酶抑制剂和抗氧化剂抑制双 tau + 毛茸茸生物测定表型的能力。我们还将测试 FDA 批准的市售化合物库（Microsource Discovery Systems）抑制 tau + 毛茸茸果蝇外眼表型的能力。随后，我们将确定已鉴定的化合物对细胞死亡以及 tau 蛋白的构象和溶解度的影响。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）将促进研究，以识别 AD 及相关病症的疾病缓解疗法。该项目将与项目 2 交互 在转基因小鼠中评估类似的化合物。