Project title: The development of bacteriophage as bionanoparticles for gene therapy
Project summary:
Gene therapy is at the forefront of innovations in healthcare. The key unmet need for transformative exploitation of gene therapy is to deliver genes to the correct tissue safely and with efficacy. Conventional viral or non-viral gene therapy approaches have considerable limitations. The intrinsic coupling of the genetics and physicochemical properties of bacteriophage makes them a unique and underexplored tool as a “Phage bioNanoParticle” (PNP) to address the challenges of gene delivery: 1) faithful targeting and efficient uptake; 2) stability in complex microenvironments with effective cargo release; 3) optimal trafficking in cells. The studentship will exploit progress in molecular screening and engineering of bacteriophage surface display systems to exploit bacteriophage as a PNP. The aim is to improve the understanding of gene delivery and provide a novel gene delivery platform.
The study will be highly novel yet based on well-established engineering principles for bacteriophage. Conventionalphage-display libraries present random peptides fused to a coat protein. The resulting PNPs display the peptide-fusion on the surface and carry packaged ssDNA coding for the peptide. This display system can be readily engineered to produce libraries with billions of peptide variants, each displaying a unique peptide and carrying the corresponding peptide gene. Panning of such libraries and subsequent Next Generation Sequencing/bioinformatics analysis of peptide genes that are within cell-associated PNPs can identify peptides with specificity for different cell types.
Study plan: 1) Targeting and Uptake: The study will design and produce a phage library-screening system to isolate proof-of-concept cell targeting peptides against model cancer cell lines (triple negative breast cancer). 2) Stability: The physicochemical properties of PNPs will be defined, and rational designs applied to facilitate effective intracellular disassembly to enhance gene transfer. 3) Trafficking: The study will combine targeting variants from steps 1-2 with established modifications to PNP coat proteins aimed to enhance endosomal escape and nuclear localisation for optimal gene expression. 4) Validation: PNPs displaying combined engineered functionalities will be fully characterised.
The research will be split between phage-display labs in the School of Veterinary Medicine and Science (SVMS, https://www.nottingham.ac.uk/vet/research/index.aspx) and cell transfection labs in the new UoN flagship research facility, the Biodiscovery Institute (BDI, https://www.nottingham.ac.uk/research/research-areas/biodiscovery-institute/biodiscovery-institute.aspx#). The academic supervisors for the project are Prof. Kevin Gough and Dr Cinzia Allegrucci from SVMS and Dr James Dixon from the School of Pharmacy (BDI). They have longstanding interests in the development of therapeutics and therapeutic delivery platforms, next generation phage display technology and cancer biology. The studentship is also supported by ADAS Biotechnology based out of their R&D labs in Beeston Business Park (Nottingham). The student will spend time at this site developing bespoke bioinformatics tools and undertaking an industry training placement. Throughout the PhD, the student will have continuous access to training in three vibrant research environments covering phage-display/molecular biology, therapeutic delivery/physical-chemical analysis methods and bioinformatics. The student will also participate in training opportunities provided by the BBSRC, host School’s and the UoN Researcher Academy.
The PhD would be suitable for students trained in Biochemistry; Bioengineering; Biology; Biotechnology; Life Sciences or Pharmacy.
To apply and check your eligibility, please click go to https://www.nottingham.ac.uk/bbdtp/apply/how-to-apply.aspx and you can find further information about how to apply to our programme.
Funding details:
Home and international students are welcome to apply for this opportunity. Funding is available for four years from late September 2023. The award covers tuition fee (£4,596) at the home rate plus an annual stipend which was (£17,668) for 2022. This is set by the Research Councils. Please note that successful international candidates will be put forward for a University Fees Difference Scholarship to cover the difference between the home and international fee.
Apply online here by noon on Tuesday 17th January 2023