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HIGH CONTENT SCREENING FOR EPITHELIAL BIOLOGY

上皮生物学的高内涵筛选

基本信息

批准号：
7356007
负责人：
LID B WONG
金额：
$ 36.14万
依托单位：
BIOTECHPLEX CORPORATION
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-09-30 至 2009-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7356007
关键词：
CONTENT SCREENING EPITHELIAL BIOLOGY

项目摘要

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）：BioTechPlex 提议开发一种高含量、高通量、基于细胞的筛选系统，用于使用上皮细胞进行药物发现。该系统将能够在 96 孔板中同时测量多个细胞荧光团可报告分子种类的动态响应。将在与各自细胞内动力学过程的时间性质一致的时间范围内测量动态响应。对三种荧光探针的动态“指纹”进行表征和分类后，将高度特异性地确定靶位点的激活或抑制。在第一阶段，BioTechPlex 开发了多荧光团声光读板器 (MAPR(tm)) 系统的光学器件和固件。 MAPR 同时测量来自 96 个细胞样品的三个荧光信号，每孔停留时间为 10 毫秒。在第二阶段，BioTechPlex将进一步开发MAPR，添加生物信息学数据库、人工智能决策并验证整个系统的制作。这将通过实现以下 3 个目标来实现。 1) 通过结合机器人板和液体处理以及温度和大气调节，为 MAPR 添加流体处理和环境调节。 2) 开发目标识别算法，对三种荧光团动态的唯一标识符进行分类、表征和归档，并提取此信息用于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、病理和工程化的呼吸道上皮细胞系验证完整的 MAPR 系统，这些信号转导途径可预测地扰乱细胞内离子，从而扰乱质膜电位。这项支持技术将为制药行业提供一个新颖的、基于多探针细胞的平台，用于高内涵筛选和高通量药物筛选，以实现药物发现、验证和细胞毒性。该系统满足 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 NCRR 为开发高通量筛选技术以及 NIDDK 为上皮生物学的普遍兴趣而提出的 2005 年 SBIR 总体征集。