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，用于使用模拟心肌的工程组织和一种新型筛选仪器进行高通量筛选试验，该仪器可以快速测量这些组织的机械性能。我们的工程组织显著减少了对整个动物试验的需求。在拟议的研究中，我们将专注于改进我们的心脏组织模型，以96孔格式生长。我们将使用我们的筛选仪器生产具有相同生理特性的样品，这些样品在高通量筛选测定中具有Z因子>0.5。我们将使用已知对心肌有影响的已知化合物进行盲法筛选，以评估我们的成功措施。此外，我们将使用已建立的毒性测定和我们的工程组织测定系统测试已知影响心血管功能的3种药物，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年的业务发展经验，将与我们合作，共同改进我们的筛选仪器，并利用我们的工程组织开发可消耗产品。在确认了该检测方法的有效性和可重复性后，我们将对菲律宾的一个小型植物提取物库进行分析。完整的提取物库（16,000个样本）只提供给威斯康星州医学院的科学家，他们合作了这个项目。NCI先前已筛选并对该库中的浸提液对HL 60细胞的细胞毒性进行了排序。该子库由1，045种浸提液组成，这些浸提液在NCI研究中显示出最低毒性。我们将通过其改变我们的组织测定系统测量的心肌收缩的2个重要参数的能力来识别“命中”提取物。在II期研究中，我们将专注于表征在我们的测定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具有成本效益地确定可以治疗心血管疾病的草药或天然产品的化合物。

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.