HIGH CONTENT SCREENING FOR EPITHELIAL BIOLOGY

上皮生物学的高内涵筛选

• 批准号: 7053673
• 负责人: LID B WONG
• 金额: $ 38.86万
• 依托单位: BIOTECHPLEX CORPORATION
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7053673
• 关键词: CONTENT; SCREENING; EPITHELIAL; BIOLOGY

DESCRIPTION (provided by applicant): BioTechPlex proposes to develop a high content, high throughput, cell-based screening system for drug discovery using epithelial cells. This system will enable the dynamic responses of multiple cellular fluorophore-reportable molecular species to be simultaneously measured in a 96 well plate. The dynamic responses will be measured in a time frame consistent with the temporal nature of the respective intracellular kinetic processes. The activation or inhibition of a target site will be determined with high specificity following the characterization and classification of the dynamic "fingerprint" from three fluorescent probes. In Phase I, BioTechPlex developed the optics and the firmware of the Multi-Fluorophore Acousto Optic Plate Reader (MAPR(tm)) system. MAPR simultaneously measures three fluorescent signals from 96 cell samples with a dwell time of 10 ms per well. In Phase II, BioTechPlex will further develop MAPR, add bioinformatics data base, Al decision making and validate the entire system making. This will be achieved by accomplishing the following 3 objectives. 1) Add fluid handling and environmental conditioning to MAPR by incorporating robotic plate and liquid handling as well as temperature and atmospheric regulation. 2) Develop Target Identification Algorithms that will classify, characterize and file the unique identifiers of the dynamics of three fluorophores as well as extract this information for target identification and decision marking. 3) Validate the complete MAPR system using normal, pathologic and engineered respiratory epithelial cell lines prior to and following perturbation of muscarinic, adrenergic and IPS receptor activated signal transduction pathways which predictably perturb intracellular ions and thus plasma membrane potential. This enabling technology will provide the pharmaceutical industry a novel, multi-probe cell-based platform for high content screening and high-throughput drug screening for drug discovery, validation, and cytotoxicity. This system addresses the needs of FDA (funded by FDA). In addition, the project directly responds to the NIH Roadmap Initiatives released in 2004. This project also directly responds to the 2005 SBIR general solicitations by NCRR for the development of high throughput screening technologies as well as by NIDDK for general interest in epithelial biology.

描述（由申请人提供）：生物技术建议使用上皮细胞开发高含量，高吞吐量，基于细胞的筛查系统，以发现药物。该系统将使多个细胞荧光团分子物种的动态响应同时在96井板中测量。动态响应将以与各个细胞内动过程的时间性质一致的时间框架进行测量。在三个荧光探针的动态“指纹”表征和分类之后，将用高特异性确定目标位点的激活或抑制。在第一阶段，Biotechplex开发了多氟化镜读取器（MAPR（TM））系统的光学和固件。 MAPR同时测量来自96个细胞样品的三个荧光信号，每孔的停留时间为10 ms。在第二阶段，Biotechplex将进一步开发MAPR，添加生物信息学数据库，AL决策并验证整个系统制造。这将通过实现以下3个目标来实现。 1）通过合并机器人板和液体处理以及温度和大气调节，将流体处理和环境条件添加到MAPR中。 2）开发目标识别算法，该算法将对三个荧光团动力学的唯一标识符进行分类，表征和提交，并提取此信息以进行目标识别和决策标记。 3）在毒蕈碱，肾上腺素能和IPS受体激活的信号转导途径扰动之前和之后，使用正常，病理和工程性呼吸道上皮细胞系验证完整的MAPR系统，这些途径可以预见，这些信号转导途径可以预见，因此可以预见，细胞内离子以及质膜膜电位。这项启示技术将为制药行业提供一个新型的，基于多探针的细胞平台，用于高素质筛查和高通量药物筛查，以发现药物发现，验证和细胞毒性。该系统解决了FDA的需求（由FDA资助）。此外，该项目直接响应2004年发布的NIH路线图计划。该项目还直接响应了NCRR 2005 SBIR一般征集，以开发高吞吐量筛选技术以及NIDDK，以及NIDDK对上皮生物学的一般兴趣。