Capsule mutations serve as a key strategy of phage resistance evolution of K54 hypervirulent Klebsiella pneumoniae

Abstract

Phage therapy is a promising antibacterial strategy against the antibiotic resistance crisis. The evolved phage resistance could pose a big challenge to clinical phage therapy. Therefore, it is necessary to conduct a comprehensive analysis of phage resistance mechanisms during treatment. Here, we characterize 37 phage-resistant mutants of hypervirulent K. pneumoniae strain SCNJ1 under phage-imposed selection in both in vitro and in vivo experiments. We show that 97.3% (36/37) of phage-resistant clones possessed at least one mutation in genes related to the CPS biosynthesis. Notably, the wcaJ gene emerges as a mutation hotspot, as mutations in this gene are detected at a high frequency under both conditions. In contrast, mutations in wzc exhibit more association with in vivo samples. These CPS-related mutants all exhibit compromised bacterial fitness and attenuated virulence in mice. Strain CM8 is the only non-CPS-related mutant, which has a bglA mutation that confers phage resistance and retains full fitness and virulence. This study highlights that laboratory characterization of phage resistance evolution can give useful insights for clinical phage therapy. This study reports phage-resistant mutants of Klebsiella pneumoniae emerging under in vitro and in vivo conditions, and demonstrates CPS mutations as the main resistance mechanism accompanied by compromised bacterial fitness and attenuated virulence.

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