High-Throughput Screening of Therapeutics

高通量表皮组织制备的可行性 模型和测试方法在I期研究中得到了证实。 在24孔板和96孔板形式中产生表皮组织。 用于HTP评估组织活力的方案和试剂， 还开发了从这些组织中分离RNA的方法。各种 用于监测基因特异性mRNA水平的定量测定是 分析了在测试的方法中，定量实时聚合酶 链反应（PCR）被认为是最通用的HTP 试验. 第二阶段的研究将进一步发展第一阶段的成果。一期 测试协议将得到优化，HTP技术将 扩张成三维器官型气管支气管组织 模型HTP EpiDerm组织模型和RNA的实用性 将通过定量实时PCR证明分离产物 血管生成基因的mRNA表达分析。此外该 HTP气管支气管模型将用于研究细胞因子 诱导调节参与气道炎症的基因， 呼吸道疾病。 拟议的商业应用： 拟议的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各种各样的 制药、化工和消费品公司， 筛选大量的候选材料。模型还将 适用于涉及基因调控的其他类型的研究 在皮肤和气道上皮中表达。

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.