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

小行星1510817 这项研究将通过同时靶向癌症特异性细胞外和细胞内线索以获得额外的特异性水平来开发一种新的癌症杀伤策略。具体而言，研究人员试图开发新一代合成DNA装置，可用于分裂自杀酶的可编程细胞内重建，该酶能够将无毒前药激活为用于杀死癌症的有毒产品。 通过将这些DNA装置与活性细胞外靶向和外源施加的非活性前药组合以触发细胞死亡，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，因此这种策略是高度可调的，并且可以针对不同的癌症标志物进行定制。教育影响包括研究生院研讨会，旨在增加研究生化学工程课程的入学人数，以及为当地高中学生和教师提供暑期实习机会。癌症治疗面临的一个主要技术障碍是如何利用癌症特异性标志物实现靶向治疗。单独的表面标记物的主动靶向是不够的，必须与细胞内信号的附加层合并以提供更高水平的特异性。我们建议通过开发一种前药治疗的变革方法来解决这一需求，该方法在复杂的细胞环境中感知细胞外和细胞内疾病状态，并启动适当的局部治疗反应用于癌症治疗。中心思想是基于支点介导的链置换来创建多层靶向、感测和响应性DNA门控锁和钥匙装置，用于分裂自杀酶的可编程细胞内重建，所述分裂自杀酶能够将前药脱氨基5-氟胞嘧啶（5-FC）活化成毒性产物5-氟尿嘧啶（5-FU）。细胞外靶向将通过与两个正交非天然氨基酸残基上的炔基和酮基位点特异性缀合来掺入PEG修饰的细胞靶向肽和内体溶解肽来实现。 蛋白质工程、合成生物学和药物输送原理的整合代表了真正的多学科努力。参与这项研究的研究生将获得连接生物化学，蛋白质工程，材料设计和药物输送的重要接口和协同作用的综合视角。该奖项由CBET部门的生物技术和生物化学工程项目共同资助，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。

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.