Tumor-Targeted Delivery of Synthetic DNA Payloads By Engineered Bacteria

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.

The 2016 UK Roadmap Bio-design for the Bio-economy highlighted the substantial impact that synthetic biology can bring to the UK and global economies by developing: frontier science and technology; establishing a healthy innovation pipeline; a highly skilled workforce and an environment in which innovative science and businesses can thrive. Synthetic biology promises to transform the UK Bio-economy landscape, bringing bio-sustainable and affordable manufacturing routes to all industrial sectors and will ensure society can tackle many contemporary global Grand Challenges including: Sustainable Manufacturing, Environmental Sustainability Energy, Global Healthcare, and Urban Development. Whilst synthetic biology is burgeoning in the UK, we now need to build on the investments made and take a further lead in training next generation scientists to ensure sustained growth of a capable workforce to underpin the science base development and growth in an advanced UK bio-economy.
This training provided by this CDT will give students from diverse backgrounds a unique synthesis of computational, biomolecular and cellular engineering skills, a peer-to-peer and industrial network, and unique entrepreneurial insight. In so doing, it will address key EPSRC priority areas and Bioeconomy strategic priorities including: Next-generation therapeutics; Engineered biomaterials; Renewable alternatives for fuels, chemicals and other small molecules; Reliable, predictable, and scalable bioprocesses; Sustainable future; Lifelong health & wellbeing.
Advances created by our BioDesign Engineering approach will address major societal challenges by delivering new routes for chemical/pharma/materials manufacture through to sustainable energy, whilst providing clean growth and reductions in energy use, greenhouse gas emissions and carbon footprints. Increased industry awareness of bio-options with better civic understanding will drive end-user demand to create market pull for products. The CDT benefits from unrivalled existing academic-industry frameworks at the host institutions, which will provide direct links to industrial partners and a direct pathway to early economic and industrial impact.

This CDT will develop 80-100 next-generation scientists and technologists (via the funded cohort and wider integration of aligned students at the three institutions) as adept scientists and engineers, instilled with technical leadership, who as broadly trained individuals will fill key skills gaps and could be expected to impact internationally through leadership roles in the medium term. Importantly the CDT addresses key skill-gaps identified with industry, which are urgently required to create and support high value jobs that will enable the UK to compete in global markets. Commercialisation and entrepreneurship training will equip the next generation of visionaries and leaders needed to accelerate and support the creation of new innovative companies to exploit these new technologies and opportunities.

The UK government identified Synthetic Biology as one of the "Eight Great Technologies" that could be a key enabler to economic and societal development. This CDT will be at the forefront of research that will accelerate the clean growth agenda and the development of a resilient circular bioeconomy, and will align with key EPSRC prosperity outcomes including a productive, healthy and resilient nation. To foster wider societal impact, the CDT will expect all students to contribute to public outreach and engagement activities including: open days, schools visits, and science festival events: students will participate in an outreach programme, with special focus on widening participation.

This CDT will contribute to the development of industrial strategy through the Synthetic Biology Leadership Council (SBLC), Industrial Biotechnology Leadership Forum (IBLF), and wider Networks in Industrial Biotechnology and Bioenergy and Professional Institutes.