Molecular responses during bacterial filamentation reveal inhibition methods of drug-resistant bacteria

Abstract

Bacterial antimicrobial resistance (AMR) is among the most significant challenges to current human society. Exposing bacteria to antibiotics can activate their self-saving responses, e.g., filamentation, leading to the development of bacterial AMR. Understanding the molecular changes during the self-saving responses can reveal new inhibition methods of drug-resistant bacteria. Herein, we used an online microfluidics mass spectrometry system for real-time characterization of metabolic changes of bacteria during filamentation under the stimulus of antibiotics. Significant pathways, e.g., nucleotide metabolism and coenzyme A biosynthesis, correlated to the filamentation of extended-spectrum beta-lactamase-producing Escherichia coli (ESBL- E. coli ) were identified. A cyclic dinucleotide, c-di-GMP, which is derived from nucleotide metabolism and reported closely related to bacterial resistance and tolerance, was observed significantly up-regulated during the bacterial filamentation. By using a chemical inhibitor, ebselen, to inhibit diguanylate cyclases which catalyzes the synthesis of c-di-GMP, the minimum inhibitory concentration of ceftriaxone against ESBL- E. coli was significantly decreased. This inhibitory effect was also verified with other ESBL- E. coli strains and other beta-lactam antibiotics, i.e., ampicillin. A mutant strain of ESBL- E. coli by knocking out the dgcM gene was used to demonstrate that the inhibition of the antibiotic resistance to beta-lactams by ebselen was mediated through the inhibition of the diguanylate cyclase DgcM and the modulation of c-di-GMP levels. Our study uncovers the molecular changes during bacterial filamentation and proposes a method to inhibit antibiotic-resistant bacteria by combining traditional antibiotics and chemical inhibitors against the enzymes involved in bacterial self-saving responses.

The global regulation of c‐di‐GMP and cAMP in bacteria

“…The activity of most DGCs/PDEs depends not only on GGDEF or EAL/HD‐GYP domains but also on signal input via an N‐terminal sensory domain 15 , 131 , 132 . Internal and external cues stimulate the N‐terminal sensory domain of DGCs and PDEs to initiate the enzyme activities that control intracellular c‐di‐GMP levels 9 , 131 , 132 , 133 , 134 . In conclusion, different signals control intracellular c‐di‐GMP levels via different DGCs/PDEs.…”

Chiral Nanoclusters as Alternative Therapeutic Strategies to Confront the Health Threat from Antibiotic-Resistant Pathogens

“…In particular, people living in developing countries are easily infected with multidrug-resistant microorganisms due to poor health services and the abuse of antibiotics. Even in the United States, the number of infections caused by antibiotic-resistant bacteria climbed from 2 million to 2.8 million within 6 years (i.e., 2013–2019). , Moreover, the USA alone spends $25 billion/year to manage chronic wounds that are associated with high rates of morbidity and mortality. − Pseudomonas aeruginosa ( P. aeruginosa ), a drug-resistant Gram-negative pathogen, can clinically cause acute or chronic infections, being one of the top-listed pathogens for hospital-acquired infections. Treatment of P. aeruginosa infection is extremely difficult because of its rapid mutations and adaptation to gain resistance to antibiotics.…”

Upregulated Palmitoleate and Oleate Production in Escherichia coli Promotes Gentamicin Resistance

“…Therefore, there is an urgent need to investigate the mechanism of antibiotic resistance in microbes. Studies reveal that metabolic reprogramming mediates antibiotic efficacy [7][8][9][10][11][12][13][14]. A metabolomic approach based on gas chromatography-mass spectrometry showed that glucose and alanine contents were decreased in kanamycin-resistant Edwardsiella tarda [15].…”

“…Studies reveal that metabolic reprogramming mediates antibiotic efficacy [ 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 ]. A metabolomic approach based on gas chromatography–mass spectrometry showed that glucose and alanine contents were decreased in kanamycin-resistant Edwardsiella tarda [ 15 ].…”