Computational Basis for On-Demand Production of Diversified Therapeutic Phage Cocktails

Mageeney, Catherine M.; Sinha, Anupama; Mosesso, Richard A.; Medlin, Douglas L.; Lau, Britney Y.; Rokes, Alecia B.; Lane, Todd W.; Branda, Steven S.; Williams, Kelly P.

doi:10.1128/mSystems.00659-20

Title: Computational Basis for On-Demand Production of Diversified Therapeutic Phage Cocktails

Journal Article · Tue Aug 11 00:00:00 EDT 2020 · mSystems

DOI:https://doi.org/10.1128/mSystems.00659-20· OSTI ID:1816411

Mageeney, Catherine M. ^[1]; Sinha, Anupama ^[1]; Mosesso, Richard A. ^[1]; Medlin, Douglas L. ^[1]; Lau, Britney Y. ^[1]; Rokes, Alecia B. ^[1]; Lane, Todd W. ^[1]; Branda, Steven S. ^[1];

^[1]

Sandia National Lab. (SNL-CA), Livermore, CA (United States)

New therapies are necessary to combat increasingly antibiotic-resistant bacterial pathogens. We have developed a technology platform of computational, molecular biology, and microbiology tools which together enable on-demand production of phages that target virtually any given bacterial isolate. Two complementary computational tools that identify and precisely map prophages and other integrative genetic elements in bacterial genomes are used to identify prophage-laden bacteria that are close relatives of the target strain. Phage genomes are engineered to disable lysogeny, through use of long amplicon PCR and Gibson assembly. Finally, the engineered phage genomes are introduced into host bacteria for phage production. As an initial demonstration, we used this approach to produce a phage cocktail against the opportunistic pathogen Pseudomonas aeruginosa PAO1. Two prophageladen P. aeruginosa strains closely related to PAO1 were identified, ATCC 39324 and ATCC 27853. Deep sequencing revealed that mitomycin C treatment of these strains induced seven phages that grow on P. aeruginosa PAO1. The most diverse five phages were engineered for nonlysogeny by deleting the integrase gene (int), which is readily identifiable and typically conveniently located at one end of the prophage. The Δint phages, individually and in cocktails, killed P. aeruginosa PAO1 in liquid culture as well as in a waxworm (Galleria mellonella) model of infection.

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Research Organization:: Sandia National Lab. (SNL-CA), Livermore, CA (United States)

Sponsoring Organization:: USDOE National Nuclear Security Administration (NNSA)

Grant/Contract Number:: NA0003525

OSTI ID:: 1816411

Journal Information:: mSystems, Vol. 5, Issue 4; ISSN 2379-5077

Publisher:: American Society for MicrobiologyCopyright Statement

Country of Publication:: United States

Language:: English

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