Host cell-based screening assays for identification of molecules targeting Pseudomonas aeruginosa cyclic di-GMP signaling and biofilm formation

Ying Hu; Jeremy Stephen Webb; Shi-Qi An

doi:10.3389/fmicb.2023.1279922

Host cell-based screening assays for identification of molecules targeting Pseudomonas aeruginosa cyclic di-GMP signaling and biofilm formation

Front Microbiol. 2023 Nov 15:14:1279922. doi: 10.3389/fmicb.2023.1279922. eCollection 2023.

Authors

Ying Hu¹, Jeremy Stephen Webb¹, Shi-Qi An¹

Affiliation

¹ National Biofilms Innovation Center and School of Biological Sciences, University of Southampton, Southampton, United Kingdom.

Abstract

The rapid emergence of bacterial resistance to antibiotics in current use is occurring worldwide and poses a significant threat to global healthcare systems. Recent research to identify new effective anti-bacterial agents has focused on regulatory pathways as targets for interference. Regulatory mechanisms employing intracellular Bis-(3',5') cyclic di-guanylate (c-di-GMP) as a secondary messenger represent a distinct category of subjects. This molecule, c-di-GMP, is present in nearly all bacterial species and plays a pivotal role in governing various biological processes, encompassing antibiotic resistance, biofilm formation, and virulence. Alteration of the cellular concentrations of the nucleotide through modulation of associated signaling pathways has the potential to reduce biofilm formation or increase susceptibility of the biofilm bacteria to antibiotics. Here, we have developed a screen for compounds that alter c-di-GMP levels in Pseudomonas aeruginosa in co-culture with bronchial epithelial cells. Through the assay of 200 natural compounds, we were able to identify several substances showing promising effects on P. aeruginosa in a host biofilm infection model. Importantly, we detected compounds that inhibit c-di-GMP levels and showed significant influence on biofilm formation and virulence in P. aeruginosa in vitro and in vivo. Consequently, we offer proof-of-concept information regarding swift and practical drug screening assays, suitable for medium- to high-throughput applications, which target the c-di-GMP signaling pathways in this significant Gram-negative pathogen.

Keywords: Pseudomonas aeruginosa; antibiotic resistance; biofilm formation; c-di-GMP; drug screening.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This study received support from the National Biofilms Innovation Center (NBIC), an Innovation and Knowledge Center funded by the Biotechnology and Biological Sciences Research Council, Innovate UK, and the Hartree Center (Award Number BB/R012415/). Additionally, funding was provided by the Wessex Medical Research (WMR) Innovation Grant (No. 519510101) to SA, and YH received a scholarship from the China Scholarship Council (CSC).