High-Throughput Screening of Therapeutics

高通量(HTP)表皮组织生产的可行性 在第一阶段研究中演示了模型和测试方法。 表皮组织以24孔板和96孔板的形式生产。 组织活力和HTP评估的方案和试剂 从这些组织中提取RNA也得到了发展。五花八门 用于监测基因特异性mRNA水平的定量分析是 分析过了。在测试的方法中，定量实时聚合酶 链式反应(PCR)被发现是HTP最通用的方法 测试。 第二阶段的研究将进一步发展第一阶段的成果。 测试协议将得到优化，HTP技术将 扩展成三维器官型气管、支气管组织 模特。HTP Epiderm组织模型和RNA的应用 分离产物将通过实时定量聚合酶链式反应进行验证 血管生成基因的mRNA表达分析。此外， HTP气管支气管模型将用于研究细胞因子 诱导调节参与呼吸道炎症的基因和 呼吸道疾病。 建议的商业应用： 提出的HTP组织模型将在产品安全方面得到应用 评估和基因调控研究的广泛种类 制药、化工和消费品公司需要 筛选出大量的候选材料。模特们还将 适用于涉及基因调控的其他类型的研究 在皮肤和呼吸道上皮细胞中表达。

The feasibility of producing high throughput (HTP) epidermal tissue models and testing methodologies was demonstrated in Phase I studies. Epidermal tissues were produced in both 24- and 96-well plate formats. Protocols and reagents for HTP assessment of tissue viability and isolation of RNA from these tissues were also developed. Various quantitative assays for monitoring gene specific mRNA levels were analyzed. Of the methods tested, quantitative real-time polymerase chain reaction (PCR) was found to be the most versatile for HTP testing. Phase II research will further develop the results of Phase I. Phase I testing protocols will be optimized and the HTP technology will be expanded to a three-dimensional organotypic tracheal bronchial tissue model. The utility of the HTP EpiDerm tissue models and RNA isolation products will be demonstrated by quantitative real-time PCR analysis of mRNA expression of angiogenesis genes. In addition, the HTP tracheal bronchial model will be used to investigate the cytokine induced regulation of genes involved in airway inflammation and respiratory diseases. PROPOSED COMMERCIAL APPLICATIONS: The proposed HTP tissue models will find application in product safety assessment and gene regulation studies for a broad variety of pharmaceutical, chemical, and consumer product companies who need to screen a large number of candidate materials. The models will also be adaptable for other types of research involving modulation of gene expression in skin and airway epithelia.