Molecular Approaches to Understanding Global Regulation

理解全球监管的分子方法

• 批准号: 6769004
• 负责人: SUSAN Celia ROBERTS
• 金额: $ 24.55万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-15 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6769004
• 关键词: Conifera; affinity chromatography; antiAIDS agent; antineoplastics; bioinformatics; biotechnology; chemical registry /resource; clone cells; complementary DNA; drug design /synthesis /production; functional /structural genomics; gene expression; gene expression profiling; genetic library; genetic regulation; genetic screening; genetically modified plants; messenger RNA; method development; paclitaxel; plants; subtraction hybridization; tissue /cell culture

DESCRIPTION (provided by applicant): Plant cell cultures can be used to alleviate the demands placed on harvest for the supply of important plant-derived medicinals. Due to economic constraints, there are currently few plant cell culture processes used commercially for the manufacture of such products. Limitations in low volumetric productivity and variability in productivity can be overcome through a better understanding of global metabolic control. Plant cell culture has received significant attention as a supply method for the anti-cancer agent paclitaxel, which has been approved by the FDA for the treatment of breast, ovarian and lung cancers as well as the AIDS-related Kaposi's sarcoma. The proposed research project centers on the use of molecular approaches to delineate global metabolic control of paclitaxel accumulation in Taxus cell cultures. We propose a significantly different approach than other research groups, one that uses transcription profiling to identify genes involved in global metabolic control. Such genes will be involved not just in paclitaxel biosynthesis (where all current efforts are focused), but also in transcriptional regulation, transport, secretion and degradation. The specific aims of this study are to (1) identify changes in gene expression in Taxus suspension cultures with enhanced accumulation of paclitaxel through the identification of mRNAs that accumulate differentially, (2) construct high quality cDNA libraries and isolate full-length clones of these differentially expressed mRNAs, (3) predict putative gene function using publicly available bioinformatics resources, and (4) functionally characterize differentially expressed genes using both stable transformation technologies and assaying of relevant enzymatic activities. The results of the proposed research will identify genes involved in global control of paclitaxel metabolism which can then be used in the design of strategies to optimize the plant cell culture process for paclitaxel production and help alleviate the paclitaxel supply problem for both patient treatments and clinical trials. It is important to emphasize that the outcomes of the proposed research, particularly the work centering around methods development, has the potential to significantly impact research on the production of other valuable medicinals from plant cell culture systems.

描述（由申请人提供）：可以使用植物细胞培养物来减轻对重要植物衍生药物供应收成的需求。由于经济限制，目前在商业上用于制造此类产品的植物细胞培养过程很少。通过更好地理解全球代谢控制，可以克服低容量生产率和生产率变化的限制。植物细胞培养已引起了抗癌剂紫杉醇的供应方法的重大关注，紫杉醇已获得FDA的批准用于治疗乳腺癌，卵巢癌和肺癌以及与艾滋病相关的Kaposi肉瘤。拟议的研究项目集中于使用分子方法来描述出租车细胞培养物中紫杉醇积累的全球代谢控制。我们提出了一种与其他研究小组的明显不同的方法，该方法使用转录分析来识别与全球代谢控制有关的基因。这种基因不仅参与紫杉醇生物合成（当前所有努力都集中在其中），还参与了转录调节，运输，分泌和降解。这项研究的具体目的是（1）确定出租车悬浮培养物中基因表达的变化，通过鉴定积累差异积累的mRNA积累增强，（2）构建高质量的cDNA库和分离型全长的克隆，并使用这些差异表达的mRNA的MRNA的MRNA的差异功能，（3）使用公共功能，使用公共功能，使用综合功能（3），（3）使用公共功能，（4转化技术和相关酶活性的分析。拟议的研究的结果将确定涉及紫杉醇代谢的全球控制的基因，然后可以在设计策略中使用，以优化紫杉醇生产的植物细胞培养过程，并有助于减轻患者治疗和临床试验的紫杉醇供应问题。重要的是要强调，拟议的研究的结果，尤其是围绕方法开发的工作，有可能显着影响植物细胞培养系统中其他有价值药物的其他有价值药物的研究。