High Throughput Profiling of Plant Extracts Using Engineered Heart Tissues

描述（由申请人提供）：该项目将优化现有的创新方法，使用模拟心肌的工程组织和一种可以快速测量这些组织的机械性能的新型筛选仪器来进行高通量筛选分析。我们的工程组织大大减少了对整个动物试验的需求。在拟议的研究中，我们将专注于完善96孔格式的心脏组织模型。我们将使用我们的筛选仪器在高通量筛选试验中生产具有相同生理特性的样品，这些样品具有z因子>0.5。我们将使用已知的对心肌有已知影响的化合物进行盲筛，以评估我们的成功措施。此外，我们将测试3种已知影响心血管功能的药物，使用既定的毒性试验和我们的工程组织试验系统并将结果关联起来。我们的战略业务合作伙伴Gilson， Inc. （Middleton， WI）在生命科学行业拥有超过50年的业务开发经验，将与我们在工程方面合作，改进我们的筛选仪器，并利用我们的工程组织开发消耗性产品。在确认该分析的有效性和可重复性后，我们将分析一个来自菲律宾的小型植物提取物库。完整的提取库（16,000个样本）仅供参与该项目的威斯康星医学院的科学家使用。NCI先前对该文库中的提取物进行了筛选并对其对HL60细胞的细胞毒性进行了排名。该子文库包括1045个在NCI研究中毒性最低的提取物。我们将通过其改变组织测定系统测量的心肌收缩的两个重要参数的能力来确定“成功”提取物。在II期研究中，我们将重点描述在我们的分析中改变心肌收缩的任何提取物中的活性成分，并筛选和分析整个文库。该项目的最终目标是快速和经济有效地从天然产品中找出一种草药或化合物，可以治疗心血管疾病。

DESCRIPTION (provided by applicant): This project will optimize an existing innovative method for performing high-throughput screening assays using engineered tissues that mimic the myocardium and a novel, screening instrument that can rapidly measure the mechanical properties of those tissues. Our engineered tissues significantly reduce the need for whole animal testing. In the proposed research, we will focus on refining our heart tissue model growing in a 96-well format. We will produce samples with identical physiological properties that have Z-factors >0.5 in high-throughput screening assays using our screening instrument. We will perform a blind screen using known compounds with known effects on myocardium to evaluate our measure of success. Further, we will test 3 drugs, known to affect cardiovascular functions, using an established toxicity assay and our engineered tissue assay system and correlate the results. Our strategic business partner, Gilson, Inc. (Middleton, WI), which has more than 50 years of business development experience in the life science industry, will collaborate with us on the engineering efforts to improve our screening instrument and develop consumable products with our engineered tissues. After confirming the usefulness and reproducibility of the assay, we will profile a small library of plant extracts from the Philippines. The full library of extracts (16,000 samples) is available only to scientists at the Medical College of Wisconsin who collaborate on this project. NCI has previously screened and ranked the extracts in this library for cytotoxicity on HL60 cells. The sub-library consists of 1,045 extracts that showed the lowest toxicity in the NCI study. We will identify "hit" extracts by their ability to alter 2 important parameters of myocardial contraction measured by our tissue assay system. In Phase II studies, we will focus on characterizing the active components in any extracts that changed myocardial contraction in our assay and screening and profiling the entire library. The ultimate goal of this project is to rapidly and cost-effectively identify an herbal remedy, or compounds from natural products, that could treat cardiovascular diseases.