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）：天然产物一直是人类丰富的资源 潜在的治疗剂。许多目前已知的天然产物是根据细胞毒性分离的。然而，有许多有机分子不具有强细胞毒性。 DNA 微阵列技术可以扩展我们发现传统筛选方法“看不见”的新化合物的能力。我们的长期目标是建立各种检测方法来发现新型治疗剂。该应用的目的是确定如何将 DNA 微阵列应用到天然产物的分离中，这是实现该目标的第一步。该应用的中心假设是转录分析可以检测细胞毒性不可见的化合物 化验。这一假设是基于并非每个基因都参与细胞增殖这一事实而提出的。拟议研究的基本原理是，一旦建立了方法论，该方法有望促进新化合物的发现。该方法甚至可以应用于已经通过其他筛选方法筛选的自然资源。在我们的方法中，DNA 微阵列用于粗混合物的初步筛选。一旦获得差异表达的基因，就可以通过更有针对性和更便宜的方法来指导活性化合物的分离，例如逆转录酶聚合酶链式反应（RT-PCR）。为了检验中心假设，我们将追求以下两个具体目标： 1. 鉴定 Keishi-bukuryo-gan（一种日本汉方药物配方，用于治疗与血瘀有关的病症）在内皮细胞中差异表达的基因簇。 2. 确定 RT-PCR 和细胞毒性指导纯化之间的差异。拟议工作的创新特点是该方法基于专门化学和生物技术的独特组合，即天然产物的微量纯化/衍生化和使用 DNA 微阵列的转录组分析。我们期望当前的研究将为有效应用基因组工具分离天然产物奠定基础。这项试点研究的结果将意义重大，因为这项研究的方法将为发现以前未被注意到的天然产物开辟新途径。