Engineering Phage Host-Range and Suppressing Bacterial Resistance through Phage Tail Fiber Mutagenesis

By Kevin Yehl, S. Lemire, Andrew C. Yang, Hiroki Ando, M. Mimee, Marcelo Der Torossian Torres, César de la Fuente-Núñez, Timothy K. Lu. 2019

370 citations recorded by the source corpus.

The rapid emergence of antibiotic-resistant infections is prompting increased interest in phage-based antimicrobials. However, acquisition of resistance by bacteria is a major issue in the successful development of phage therapies. Through natural evolution and structural modeling, we identified host-range determining regions (HRDR) in the T3 phage tail fiber protein and developed a high-throughput strategy to genetically engineer these regions through site-directed mutagenesis. Inspired by antibody specificity engineering, this approach generates deep functional diversity (>107 different members), while minimizing disruptions to the overall protein structure, resulting in synthetic “phagebodies”. We showed that mutating HRDRs yields phagebodies with altered host-ranges. Select phagebodies enable long-term suppression of bacterial growth by preventing the appearance of resistance in vitro and are functional in vivo using a mouse skin infection model. We anticipate this approach may facilitate the creation of next-generation antimicrobials that slow resistance development and could be extended to other viral scaffolds for a broad range of applications. Highlights Vastly diverse phagebody libraries containing 107 different members were created. Structure-informed engineering of viral tail fibers efficiently generated host-range alterations. Phagebodies prevented the development of bacterial resistance across long timescales in vitro and are functional in vivo.

Subfields: Ecology, Microbiology, Radiology, Nuclear Medicine and Imaging

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Citations in the corpus

Pseudomonas aeruginosa: pathogenesis, virulence factors, antibiotic resistance, interaction with host, technology advances and emerging therapeutics (2022, background): “Notably, LPS mutations confer bacteria gain tolerance to phage infection.(41) Taken together, in addition to the direct interaction with the host PRRs receptors, LPS may use its unique molecular features to adjust bacterial pathogenesis and damage host immune defense, ultimately benefiting the fitne”