Tumor-Targeted Delivery of Synthetic DNA Payloads By Engineered Bacteria

通过工程细菌靶向肿瘤递送合成 DNA 有效负载

• 批准号: 2602443
• 金额: --
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2602443
• 关键词: Tumor; Targeted; Delivery; Synthetic; DNA

Bacteria capable of invasion into human cells offer a promising technology for cell-based therapies and could also be used to provide a route for DNA delivery, potentially enabling these bacteria to be a vector for gene therapies, DNA-based vaccines and in vivo genome engineering. Past work has demonstrated how modular DNA constructs can be used to engineer E. coli and Salmonella to target and enter cancer cell lines and tumours and specifically deliver protein payloads (e.g. toxins). The proposed project will go beyond delivery of just proteins and develop a platform for using tumor-targeting bacteria for the controlled delivery of DNA payloads that are then expressed in the host cell. We anticipate that this can become a core technology for both bacterial cell-based therapies and for DNA transfer for mammalian synthetic biology and synthetic genomics.This interdisciplinary project will bring together the foundational synthetic biology methods of DNA-based engineering and design of synthetic expression (Tom Ellis) with cutting-edge biomedical research of intracellular host-microbe interactions (Teresa Thurston, Ramesh Wigneshweraraj). It is co-sponsored by an exciting London-based industrial partner, Prokarium, and leverages their expertise and commercial knowledge of bacterial cell therapies. The research will build on an existing synthetic biology project between the company and the London BioFoundry, but now take this in the new direction of DNA delivery.The planned experimental work will see the student design, construct and test modular DNA programs that trigger secretion mechanisms and conjugation in relevant E. coli and Salmonella strains when these bacteria enter target cells. A core aim of this project will be to use non-pathogenic Salmonella strains as a vehicle to inject DNA encoding a eukaryotic gene into a cancer cell line. This process will be optimised and measured by tracking DNA expression and location by fluorescent imaging and single-molecule microscopy. Further work will look to test the limit of the lengths of DNA that can be transferred. Conjugation between bacteria is regularly used to transfer millions of bases of DNA in one go, and so this offers promise as a route to transfer big sections of synthetic DNA into target cells.

能够侵入人体细胞的细菌为基于细胞的治疗提供了一种有前景的技术，也可用于提供 DNA 传递途径，从而有可能使这些细菌成为基因治疗、基于 DNA 的疫苗和体内基因组工程的载体。过去的工作已经证明了如何使用模块化 DNA 构建体来改造大肠杆菌和沙门氏菌，以靶向并进入癌细胞系和肿瘤，并特异性地传递蛋白质有效负载（例如毒素）。拟议的项目将不仅仅是蛋白质的递送，还将开发一个平台，利用肿瘤靶向细菌来控制 DNA 有效负载的递送，然后在宿主细胞中表达。我们预计这可以成为基于细菌细胞的疗法以及哺乳动物合成生物学和合成基因组学的 DNA 转移的核心技术。这个跨学科项目将结合基于 DNA 的工程和合成表达设计的基础合成生物学方法（Tom Ellis）与细胞内宿主-微生物相互作用的尖端生物医学研究（Teresa Thurston、Ramesh） 维涅什维拉杰）。它由伦敦一家令人兴奋的工业合作伙伴 Prokarium 共同赞助，并利用他们在细菌细胞疗法方面的专业知识和商业知识。该研究将建立在该公司与 London BioFoundry 之间现有的合成生物学项目的基础上，但现在将其转向 DNA 传递的新方向。计划中的实验工作将让学生设计、构建和测试模块化 DNA 程序，当这些细菌进入目标细胞时，这些程序会触发相关大肠杆菌和沙门氏菌菌株的分泌机制和结合。该项目的核心目标是使用非致病性沙门氏菌菌株作为载体，将编码真核基因的 DNA 注入癌细胞系。该过程将通过荧光成像和单分子显微镜追踪 DNA 表达和位置来优化和测量。进一步的工作将测试可转移 DNA 长度的极限。细菌之间的接合通常用于一次性转移数百万个 DNA 碱基，因此这有望成为将大段合成 DNA 转移到靶细胞中的途径。