Project Outline

Bacteriophages are viruses with great specificity for infecting their bacterial hosts, allowing their use as targeted antibacterial agents. Phage therapy has received more and more interest nowadays, due to the problem of antimicrobial resistance. Recently, the UK government released a Commons Science, Innovation and Technology Committee inquiry on the antimicrobial potential of bacteriophages (https://committees.parliament.uk/call-for-evidence/2970/), indicating the urgency of phage therapy to be established in UK.

In this project, we propose to isolate, characterize and study the mechanisms of action of novel bacteriophages targeting clinical E.coli K1 isolates. Sagona lab has worked extensively on K1F bacteriophage (1,2,3,4) and its respective bacterial hosts and this knowledge will be used to expand now on clinical isolates and the novel phages targeting those.

Aims and Objectives

The overarching aim of the project is to isolate, characterize and study the mechanisms of action of novel E.coli K1 Bacteriophages towards their clinical host.

The specific aims of the project are the following:

1. To receive in collaboration with UCWH hospital clinicians, clinical bacterial E.coli samples, sequence those, characterize those for antibiotic susceptibility and store those.
2. To isolate based on the clinical isolates novel bacteriophages that can efficiently target the E.coli clinical isolates.
3. To sequence, characterize using microbiological bioinformatic assays the novel bacteriophages.
4. To perform advanced electron microscopy to identify the structure of these bacteriophages.
5. To perform biochemical and cell biology assays to understand better the mechanisms via which these bacteriophages can clear E.coli infection in human cell environment.

A combination of microbiology, phage biology, biochemistry, electron microscopy, bioinformatics human cell biology and microscopy methods, will enable us to reach the objectives of the project.

References

1. Engineered K1F bacteriophages kill intracellular Escherichia coli K1 in human epithelial cells. Møller-Olsen C, Ho SFS, Shukla RD, Feher T, Sagona AP.Sci Rep. 2018 Dec 3;8(1):17559. doi: 10.1038/s41598-018-35859-6.PMID: 30510202
2. Genetic Engineering of Bacteriophage K1F with Human Epidermal Growth Factor to Enhance Killing of Intracellular E. coli K1.Williams J, Kerven J, Chen Y, Sagona AP.ACS Synth Biol. 2023 Jul 21;12(7):2094-2106. doi: 10.1021/acssynbio.3c00135. Epub 2023 Jun 15.PMID: 37318278
3. Transposable Element Insertions into the Escherichia coli Polysialic Acid Gene Cluster Result in Resistance to the K1F Bacteriophage.Styles KM, Locke RK, Cowley LA, Brown AT, Sagona AP.Microbiol Spectr. 2022 Jun 29;10(3):e0211221. doi: 10.1128/spectrum.02112-21. Epub 2022 Apr 25.PMID: 35467398
4. Bacteriophage K1F targets Escherichia coli K1 in cerebral endothelial cells and influences the barrier function. Møller-Olsen C, Ross T, Leppard KN, Foisor V, Smith C, Grammatopoulos DK, Sagona AP.Sci Rep. 2020 Jun 1;10(1):8903. doi: 10.1038/s41598-020-65867-4.PMID: 32483257

Techniques

Microbiology, phage biology, biochemistry, advanced electron microscopy and structure biology, bioinformatics, human cell biology and microscopy methods, will be used in this project.