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心脏病的主要促成因素。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，但仍然需要改进药物，减少副作用。该提案的目标是开发一种荧光斑马鱼测定法（Z-Lipotrack），以鉴定用于脂质管理的新药。斑马鱼作为脊椎动物疾病的模型具有许多优势。特别是，它们以类似于哺乳动物的方式通过消化系统处理脂质。由于斑马鱼幼虫基本上是透明的，因此在荧光脂质底物的帮助下，可以在整个生物体中容易地观察到这种加工，所述荧光脂质底物被斑马鱼幼虫吞下并从肠运输到肝脏和胆囊。为了提高Z-Lipotrack在高通量药物筛选中的有用性，将开发荧光酶标仪，以自动对Z-Lipotrack检测进行成像。其次，将评价Z-Lipotrack用于化合物优化的使用。最后，在第1阶段确定为潜在降脂药物的化合物将通过使用药物化学进行充分表征和优化，以确定进一步动物和人体研究的先导化合物。

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.