Screening Device for Differentiated Primary Cell Models of Airway Epithelia

气道上皮分化原代细胞模型筛选装置

基本信息

批准号：
8515510
负责人：
Robert G Lowery
金额：
$ 19.4万
依托单位：
BELLBROOK LABS, LLC
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8515510
关键词：
Academia Air Animal Testing Biological Assay Biological Availability Biological Process Biomedical Research Blood - brain barrier anatomy Cell Line Cell model Cells Chronic Chronic Obstructive Airway Disease Consumption Cosmetics Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Devices Disease Disease model Drug Transport Electrophysiology (science)Epithelial Feasibility Studies Genetic Engineering Goals Gold Growth Height Human Image Industry Inflammatory Inflammatory Response Investments Ion Channel Kidney Diseases Liquid substance Lung Lung diseases Marketing Microscopy Modeling Nose Optics Oral Other Genetics Permeability Pharmaceutical Preparations Play Polycystic Kidney Diseases Preclinical Drug Evaluation Price Proteins Reagent Reporter Research Resolution Respiratory Tract Infections Robotics Role Savings Skin Skin Cancer Solutions System Techniques Technology Testing Therapeutic Tissues Vitelliform macular dystrophy airway epithelium airway surface liquid cost cytokine density design drug candidate drug development drug discovery evaporation eye dryness high throughput screening human disease immortalized cell immunocytochemistry improved instrument interest meetings miniaturize novel operation overexpression programs prototype public health relevance response screening success tool trafficking

项目摘要

DESCRIPTION (provided by applicant): Approaches to drug screening typically involve dramatic compromises in order to achieve high throughput and hold down costs. Examples include the use of immortalized, heterologous cell lines, and genetic engineering to over-express the target protein and/or incorporate a fluorescent surrogate reporter to display the result. At least partly because such systems often produce responses that are not relevant to the actual human disease state, high throughput screening has not historically delivered a strong return on investment. More representative primary cell models are often available for tissues and diseases, but these models are underutilized in screening because of the lack of technological solutions and high costs. For similar reasons, more relevant assay technologies such as endogenous immunocytochemistry and electrophysiology are not commonly employed in HTS. The goal of this project is to bring the gold standard, organotypic cell model for airway epithelia into true high throughput screening, and enable the use of more informative high content assays including endogenous CFTR trafficking and airway surface liquid height. In addition to applications for cystic fibrosis and COPD, the resultant device will also be valuable for skin models and drug transport studies. The device consists of an array of 96 microchambers in standard microplate format. The microchamber design is small enough to be compatible with 384-well plate densities, so 384 microchambers per plate is readily achievable. The plate includes specialized features for compatibility and ease-of-use with standard liquid handling robotics, and high resolution microscopy. Importantly, the device is simple enough to be produced at a cost consistent with the cost constraints of HTS labs, and confers dramatic savings in primary cell and media consumption. The feasibility study on a small-scale device was successfully completed, indicating that the highly miniaturized approach is compatible with organotypic epithelial airway culture at air-liquid interface. Although not an aim of this proposal this device is designed to be integrated into an electrophysiology instrument designed specifically for this culture system.

描述（由申请人提供）：药物筛选方法通常涉及显着的妥协，以实现高通量并降低成本。例子包括使用永生化异源细胞系和基因工程来过度表达靶蛋白和/或掺入荧光替代报告基因以显示结果。至少部分原因是此类系统经常产生与实际人类疾病状态无关的反应，因此高通量筛选历史上并未带来丰厚的投资回报。更具代表性的原代细胞模型通常可用于组织和疾病，但由于缺乏技术解决方案和高昂的成本，这些模型在筛选中并未得到充分利用。出于类似的原因，HTS 中并不普遍采用更相关的检测技术，例如内源免疫细胞化学和电生理学。该项目的目标是将气道上皮的金标准、器官型细胞模型引入真正的高通量筛选，并能够使用信息更丰富的高内涵检测，包括内源性 CFTR 运输和气道表面液体高度。除了囊性纤维化和慢性阻塞性肺病的应用之外，所得设备对于皮肤模型和药物转运研究也很有价值。该装置由标准微孔板格式的 96 个微室阵列组成。微室设计足够小，可与 384 孔板密度兼容，因此每个板可轻松实现 384 个微室。该板具有与标准液体处理机器人和高分辨率显微镜兼容且易于使用的专门功能。重要的是，该设备足够简单，生产成本符合 HTS 实验室的成本限制，并且可以大大节省原代细胞和培养基的消耗。小型装置的可行性研究已成功完成，表明高度小型化的方法与气液界面的器官型上皮气道培养兼容。尽管不是本提案的目的，但该设备旨在集成到专门为此培养系统设计的电生理学仪器中。