Zebrafish Lipid Metabolism Assay for Drug Screening

描述(申请人提供)：心血管疾病是当今美国的头号死因。高水平的胆固醇和甘油三酯已被确定为心脏病的主要致病因素。此外，他汀类和贝特类等降脂药物已被证明可以降低冠状动脉疾病的发生率。尽管目前的药物在这一领域取得了一些成功，但仍然需要副作用更少的改进药物。这项建议的目标是开发一种荧光斑马鱼试验(Z-LipoTrack)，以确定用于血脂管理的新药。斑马鱼在模拟脊椎动物疾病方面有许多优势。特别是，它们通过消化系统处理脂肪的方式与哺乳动物类似。由于斑马鱼幼体基本上是透明的，在荧光脂类底物的帮助下，这种过程很容易在整个生物体中观察到，这些底物被斑马鱼幼体吞噬，并从肠道运输到肝脏和胆汁。为了提高Z-LipoTrack在高通量药物筛选中的有效性，将开发一种荧光平板阅读器来自动对Z-LipoTrack分析进行成像。其次，对Z-LipoTrack在配方优化中的应用进行了评价。最后，在第一阶段中被确定为潜在降脂药物的化合物将通过使用药物化学来识别先导化合物以供进一步的动物和人类研究来充分表征和优化。

DESCRIPTION (provided by applicant): Cardiovascular disease is the number one cause of death in the United States today. High levels of cholesterol and triglycerides have been identified as major contributing factors to heart disease. Furthermore, lipid-lowering drugs such as statins and fibrates have been shown to reduce the incidence of coronary disease. Although current drugs have enjoyed some success in this area, a need remains for improved drugs with fewer side effects. The goal of this proposal is to develop a fluorescent zebrafish assay (Z-Lipotrack) to identify new drugs for lipid management. The zebrafish has many advantages to model vertebrate diseases. In particular, they process lipids through the digestive system in a manner similar to mammals. Because zebrafish larvae are essentially transparent, such processing can be readily observed in the whole organism, with the aid of fluorescent lipid substrates, which are swallowed by the zebrafish larvae and transported from the intestine to the liver and gall bladder. To increase the usefulness of Z-Lipotrack for high throughput drug screening, a fluorescent plate reader will be developed to automatically image the Z-Lipotrack assay. Secondly, the use of Z-Lipotrack for compound optimization will be evaluated. Finally, compounds identified as potential lipid-lowering drugs in Phase 1will be fully characterized and optimized through the use of medicinal chemistry to identify lead compounds for further animal and human studies.