Zebrafish Lipid Metabolism Assay for Drug Screening

描述（由申请人提供）：心血管疾病是当今美国的头号死亡原因。高水平的胆固醇和甘油三酯已被确定为导致心脏病的主要因素。此外，降脂药物如他汀类药物和贝特类药物已被证明可以降低冠心病的发病率。虽然目前的药物在这方面取得了一些成功，但仍然需要改进副作用更小的药物。本提案的目标是开发一种荧光斑马鱼测定法（Z-Lipotrack），以识别用于脂质管理的新药。斑马鱼在模拟脊椎动物疾病方面有许多优点。特别是，它们通过消化系统处理脂质的方式与哺乳动物相似。由于斑马鱼幼虫本质上是透明的，在荧光脂质底物的帮助下，可以很容易地在整个生物体中观察到这种加工过程，这些底物被斑马鱼幼虫吞下，并从肠道运输到肝脏和胆囊。为了提高Z-Lipotrack在高通量药物筛选中的实用性，将开发一种荧光板阅读器来自动成像Z-Lipotrack试验。其次，对Z-Lipotrack在复方优化中的应用进行了评价。最后，通过药物化学鉴定先导化合物进行进一步的动物和人体研究，充分表征和优化在i期被确定为潜在降脂药物的化合物。

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.