Transcriptional Control of Alkaloid Biosynthesis in Catharanthus roseus Cultures

这项由美国国家科学基金会颁发的生物技术、生物化学和生物质工程项目的奖项支持以下研究：1)更好地理解细胞如何调节马达加斯加长春花植物（Catharanthus roseus）培养的药用生物碱的生产；2)使用基于基因沉默的新策略来提高这些关键药物化合物的生产。知识价值：本项目将提高红花培养物中萜类吲哚生物碱（TIAs）的产量。玫瑰属植物生产多种高价值药物，包括抗癌药物长春新碱和长春碱，但植物生长速度慢，产品浓度低是有效生产药物的重大障碍。这些药物的高成本和需求促使我们的研究更好地了解它们的生物合成，并最终利用玫瑰孢杆菌培养物过量生产这些化合物。提出的研究目标是描述TIA生物合成所需的酶的转录调控，并设计一种有效的工程策略来克服TIA生物合成的先天障碍。本研究的具体目的是：1)开发具有转录调控因子表达改变的转基因玫瑰花毛状根培养物；2)表征TIA生物合成相关基因的生物碱谱和转录谱；3)深入了解TIA产生的分子机制。了解TIA生物合成的转录调控将为新的基因工程策略提供基础，以提高玫瑰花培养物的TIA产量。更广泛的影响：本研究的愿景是解决使用植物细胞培养的关键植物衍生药物的替代供应需求。从植物细胞培养物中提高这些药物的体积生产力将降低患者的药物成本，并使药物的供应可靠。此外，PI/Co-PI将1)将他们研究中的概念和例子纳入他们的课程，并开发新的实验室模块；2)共同指导研究生和本科生进行跨学科研究，将生化工程、基因工程和分子生物学方法应用于这项研究；3)通过让教师和学生在他们的实验室进行实践研究经验来支持K-12 STEM教育。

This NSF award by the Biotechnology, Biochemical, and Biomass Engineering program supports research to 1) better understand how the cell regulates the production of medicinal alkaloids from cultures of the Madagascar periwinkle plant (Catharanthus roseus) and 2) use a new strategy based on gene silencing to enhance the production of these critical pharmaceutical compounds. Intellectual Merit: This project will improve the production of terpenoid indole alkaloids (TIAs) in cultures of Catharanthus roseus. C. roseus produces several highly-valued pharmaceuticals, including the anti-cancer drugs vincristine and vinblastine, but the slow growth rate of the plant and the low concentration of product are significant barriers to efficient drug production. The high cost and need for these pharmaceuticals motivate our research to better understand their biosynthesis and ultimately overproduce these compounds using C. roseus cultures. The goals of the proposed research are to describe the transcriptional regulation of enzymes required for TIA biosynthesis and to design an effective engineering strategy for overcoming innate blocks to TIA biosynthesis. The specific aims of this research are 1) to develop transgenic C. roseus hairy root cultures with altered expression of transcriptional regulators, 2) to characterize the alkaloid profiles and transcriptional profiles of genes relevant to TIA biosynthesis, and 3) to gain insight into the molecular mechanism of TIA production. Understanding the transcriptional regulation of TIA biosynthesis will provide the basis for new genetic engineering strategies for enhancing TIA production from C. roseus cultures. Broader Impact: The vision of this research is to address the need for an alternative supply of critical plant-derived pharmaceuticals using plant cell cultures. Increasing the volumetric productivity of these pharmaceuticals from plant cell cultures will reduce the cost of the drug to the patient and enable a dependable supply of the drug. In addition, the PI/Co-PI will 1) incorporate concepts and examples from their research into their courses and develop new lab modules, 2) co-advise graduate students and undergraduates in interdisciplinary research that applies biochemical engineering, genetic engineering, and molecular biology approaches to this research, and 3) support K-12 STEM education by hosting teachers and students through hands-on research experiences in their laboratory.