sRNA expression profile of KPC-2-producing carbapenem-resistant Klebsiella pneumoniae: Functional role of sRNA51

PLoS Pathog. 2024 May 8;20(5):e1012187. doi: 10.1371/journal.ppat.1012187. eCollection 2024 May.

Abstract

The emergence of carbapenem-resistant Klebsiella pneumoniae (CRKP) has significant challenges to human health and clinical treatment, with KPC-2-producing CRKP being the predominant epidemic strain. Therefore, there is an urgent need to identify new therapeutic targets and strategies. Non-coding small RNA (sRNA) is a post-transcriptional regulator of genes involved in important biological processes in bacteria and represents an emerging therapeutic strategy for antibiotic-resistant bacteria. In this study, we analyzed the transcription profile of KPC-2-producing CRKP using RNA-seq. Of the 4693 known genes detected, the expression of 307 genes was significantly different from that of carbapenem-sensitive Klebsiella pneumoniae (CSKP), including 133 up-regulated and 174 down-regulated genes. Both the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment and Gene Ontology (GO) analysis showed that these differentially expressed genes (DEGs) were mainly related to metabolism. In addition, we identified the sRNA expression profile of KPC-2-producing CRKP for the first time and detected 115 sRNAs, including 112 newly discovered sRNAs. Compared to CSKP, 43 sRNAs were differentially expressed in KPC-2-producing CRKP, including 39 up-regulated and 4 down-regulated sRNAs. We chose sRNA51, the most significantly differentially expressed sRNA in KPC-2-producing CRKP, as our research subject. By constructing sRNA51-overexpressing KPC-2-producing CRKP strains, we found that sRNA51 overexpression down-regulated the expression of acrA and alleviated resistance to meropenem and ertapenem in KPC-2-producing CRKP, while overexpression of acrA in sRNA51-overexpressing strains restored the reduction of resistance. Therefore, we speculated that sRNA51 could affect the resistance of KPC-2-producing CRKP by inhibiting acrA expression and affecting the formation of efflux pumps. This provides a new approach for developing antibiotic adjuvants to restore the sensitivity of CRKP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Carbapenem-Resistant Enterobacteriaceae* / genetics
  • Carbapenems / pharmacology
  • Gene Expression Regulation, Bacterial
  • Klebsiella Infections / drug therapy
  • Klebsiella Infections / genetics
  • Klebsiella Infections / microbiology
  • Klebsiella pneumoniae* / drug effects
  • Klebsiella pneumoniae* / genetics
  • Klebsiella pneumoniae* / metabolism
  • Microbial Sensitivity Tests
  • RNA, Bacterial* / genetics
  • RNA, Small Untranslated* / genetics
  • beta-Lactamases* / genetics
  • beta-Lactamases* / metabolism

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • beta-lactamase KPC-2, Klebsiella pneumoniae
  • beta-Lactamases
  • Carbapenems
  • RNA, Bacterial
  • RNA, Small Untranslated

Grants and funding

This work was supported by the Special Foundation for National Science and Technology Basic Research Program of China (Grants 2019 FY101200 to Y.L.), and Shengjing Hospital of China Medical University (“345 talent project” to Y.B.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.