Genomic Screening of Kampo Herbal Medicine (pilot)

汉方药材基因组筛选（试点）

• 批准号: 6773135
• 负责人: AKIRA KAWAMURA
• 金额: $ 3.4万
• 依托单位: HUNTER COLLEGE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6773135
• 关键词: alternative medicine; biotechnology; drug discovery /isolation; folk medicine; gene expression profiling; genetic screening; medicinal plants; microarray technology; plant extracts; polymerase chain reaction

DESCRIPTION (provided by applicant): Natural products have been serving as rich resources of potential therapeutic agents. Many of the presently known natural products were isolated based on cytotoxicity. There are, however, a number of organic molecules that are devoid of strong cytotoxicity. DNA microarray technology can expand our ability to uncover novel compounds that have been 'invisible' to the conventional screening methods. Our long-range goal is to establish various assay methodologies for the discovery of novel therapeutic agents. The objective of this application, which is the first step in pursuit of that goal, is to determine how DNA microarray can be implemented into the isolation of natural products. The central hypothesis of this application is that transcription profiling can detect compounds that are invisible to cytotoxicity assay. This hypothesis was formulated based on the fact that not every gene is involved in the cell proliferation. The rationale for the proposed research is that, once the methodology is established, the approach is expected to facilitate the discovery of novel compounds. The method can be applied even to natural resources that have already been screened by other screening methods. In our approach, DNA microarray is employed for initial screening of crude mixtures. Once differentially expressed genes are obtained, the isolation of active compounds can be guided by more focused and less expensive methods, such as reverse-transcriptase polymerase chain reaction (RT-PCR). In order to test the central hypothesis, the following two specific aims will be pursued: 1. Identify clusters of genes differentially expressed in endothelial cells by Keishi-bukuryo-gan, a Japanese Kampo medicine formulation for conditions associated with blood stasis. 2. Determine the differences between RT-PCR and cytotoxicity-guided purification. The innovative feature of the proposed work is that the approach is based on a unique combination of specialized chemical and biological techniques, namely, microscale purification/ derivatization of natural products and transcriptome profiling using DNA microarray. It is our expectation that the current study will found the basis for an effective application of genomic tools for natural products isolation. The results from this pilot study will be significant, because the methods coming out of this study will open up new avenues for the discovery of previously unnoticed natural products.

描述(申请人提供)：天然产品一直是丰富的资源 潜在的治疗剂。目前已知的许多天然产物都是基于细胞毒性而分离出来的。然而，有一些有机分子没有很强的细胞毒性。DNA微阵列技术可以扩大我们发现传统筛选方法无法发现的新化合物的能力。我们的长期目标是建立各种检测方法，以发现新的治疗药物。这项应用的目标是确定如何将DNA微阵列应用于天然产品的分离，这是追求这一目标的第一步。这一应用的中心假设是，转录图谱可以检测到对细胞毒性不可见的化合物 化验。这一假说是基于并不是每个基因都参与细胞增殖这一事实而提出的。拟议研究的理由是，一旦建立了方法学，这种方法有望促进新化合物的发现。这种方法甚至可以应用于已经通过其他筛选方法进行筛选的自然资源。在我们的方法中，DNA微阵列被用于粗混合物的初步筛选。一旦获得差异表达的基因，就可以用更有针对性和更便宜的方法来指导活性化合物的分离，例如逆转录聚合酶链式反应(RT-PCR)。为了验证这一中心假说，我们将追求以下两个具体目标：1.确定在血管内皮细胞中差异表达的基因簇，这是一种治疗血瘀症的日本Kampo药物配方。2.比较RT-PCR和细胞毒性导向纯化方法的差异。拟议工作的创新特点是，该方法基于专门的化学和生物技术的独特组合，即自然产物的微尺度纯化/衍生化和使用DNA微阵列的转录组分析。我们期望目前的研究将为基因组工具在天然产物分离中的有效应用奠定基础。这项初步研究的结果将是重要的，因为这项研究产生的方法将为发现以前未被注意到的天然产品开辟新的途径。