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

这项研究将通过同时靶向癌症特异性的细胞外和细胞内信号以获得更高水平的特异性来开发一种新的癌症杀伤策略。具体来说，研究人员寻求开发新一代合成DNA装置，该装置可用于分裂自杀酶的可编程细胞内重构，该酶能够将无毒的前药激活为有毒的抗癌产物。通过将这些DNA装置与活性细胞外靶向和外源性应用非活性前药相结合来触发细胞死亡，可以实现对癌细胞更高程度的特异性。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，因此该策略具有高度可调性，并且可以针对不同的感兴趣的癌症标志物进行定制。教育方面的影响包括研究生院研讨会，旨在增加化学工程研究生项目中代表性不足的学生的入学率，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来实现靶向治疗。单独的表面标记物的主动靶向是不够的，必须与额外的细胞内信号层合并，以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解决这一需求，该方法可以在复杂的细胞环境中感知细胞外和细胞内疾病状态，并为癌症治疗启动适当的局部治疗反应。核心思想是基于脚点介导的链位移，为分裂自杀酶的可编程细胞内重构创造多层靶向、传感和响应性dna门控锁和钥匙装置，该酶能够激活前药脱胺酸5-氟胞嘧啶（5-FC）转化为有毒产物5-氟尿嘧啶（5-FU）。细胞外靶向将通过结合peg修饰的细胞靶向和内溶肽，通过与两个正交的非天然氨基酸残基上的炔基和酮基进行位点特异性偶联来实现。蛋白质工程、合成生物学和药物传递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一个综合的视角，了解生物化学、蛋白质工程、材料设计和药物传递之间的重要界面和协同作用。该奖项由CBET部门的生物技术和生化工程项目颁发，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.