UNS:Synthetic multilayer targeting DNA devices for detection of specific cancer indicators and programmed assembly of split yCD for prodrug activation

UNS：用于检测特定癌症指标的合成多层靶向 DNA 装置和用于前药激活的分裂 yCD 的程序组装

• 批准号: 1510817
• 负责人: Wilfred Chen
• 金额: $ 45万
• 依托单位: University of Delaware
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1510817&HistoricalAwards=false
• 关键词: UNS; Synthetic; multilayer; targeting; DNA

1510817 Chen, Wilfred This research will develop a new cancer-killing strategy by simultaneously targeting cancer-specific extracellular and intracellular cues for additional levels of specificity. Specifically, the investigators seek to develop a new generation of synthetic DNA devices that can be used for programmable intracellular reconstitution of a split suicide enzyme capable of activating a non-toxic prodrug into a toxic product for cancer killing. By combining these DNA devices with active extracellular targeting and exogenously applied, inactive prodrugs to trigger cell death, a higher degree of specificity for cancer cells can be achieved. Because each of these components can be independently designed to achieve the desired outputs, this strategy is highly tunable and can be customized for different cancer markers of interest. Educational impacts include graduate school workshops aimed at increasing enrollment of underrepresented students in graduate Chemical Engineering programs and summer internships for local high school students and teachers.A major technological hurdle confronting cancer therapeutics is how to take advantage of cancer-specific markers to achieve targeted therapy. Active targeting of surface markers alone is inadequate and must be merged with additional layers of intracellular signals to provide a higher level of specificity. We propose to address this need by developing a transformative approach of prodrug therapy that senses both the extracellular and intracellular disease states in a complex cellular environment and actuates an appropriate, localized therapeutic response for cancer treatment. The central idea is to create multi-layer targeting, sensing, and responsive DNA-gated lock and key devices based on toehold-mediated strand displacement for programmable intracellular reconstitution of a split suicide enzyme capable of activating the prodrug deaminate 5-fluorocytosine (5-FC) into the toxic product 5-fluorouracil (5-FU). Extracellular targeting will be achieved by incorporating PEG-modified cell-targeting and endosomolytic peptides via site-specific conjugation with the alkyne and keto groups on two orthogonal unnatural amino acid residues. The integration of principles from protein engineering, synthetic biology, and drug delivery represents a truly multidisciplinary effort. Graduate students participating in this research will gain an integrated perspective of the important interfaces and synergies connecting biochemistry, protein engineering, material design, and drug delivery.This award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the Systems and Synthetic Biology Program of the Division of Molecular and Cellular Biology.

1510817陈，威尔弗雷德这项研究将开发一种新的癌症杀灭策略，通过同时针对癌症特定的细胞外和细胞内线索来获得更高水平的特异性。具体地说，研究人员试图开发新一代合成DNA设备，这种设备可以用于可编程地在细胞内重组分裂的自杀酶，从而能够将无毒的前体药物激活为有毒的产品，从而杀死癌症。通过将这些DNA设备与主动的细胞外靶向和外源应用的非活性前体药物触发细胞死亡相结合，可以实现对癌细胞的更高程度的特异性。由于这些组件中的每一个都可以独立设计以实现所需的输出，因此该策略是高度可调的，可以针对不同的感兴趣的癌症标记物进行定制。教育影响包括旨在增加化学工程研究生项目招生人数的研究生院讲习班，以及当地高中生和教师的暑期实习。癌症治疗学面临的一个主要技术障碍是如何利用癌症特异性标记物实现靶向治疗。仅仅主动靶向表面标志物是不够的，必须与额外的细胞内信号层合并才能提供更高水平的特异性。我们建议通过开发一种变革性的前药物治疗方法来满足这一需求，这种方法可以在复杂的细胞环境中感知细胞外和细胞内的疾病状态，并对癌症治疗产生适当的、局部的治疗反应。其中心思想是创建多层靶向、传感和响应DNA门控锁定和基于脚趾介导的链置换的关键设备，用于可编程地在细胞内重组分裂的自杀酶，该酶能够激活前药脱氨基5-氟胞嘧啶(5-FC)为有毒产品5-氟尿嘧啶(5-FU)。通过与两个相互垂直的非天然氨基酸残基上的炔基和酮基进行定点结合，将聚乙二醇化的细胞靶向和内溶肽结合到胞外靶向。蛋白质工程、合成生物学和药物输送原理的整合代表了一项真正的多学科努力。参加这项研究的研究生将对连接生物化学、蛋白质工程、材料设计和药物输送的重要接口和协同作用有一个综合的看法。这个奖项由CBET部门的生物技术和生化工程项目获得，由分子和细胞生物学部门的系统和合成生物学项目共同资助。