Effects of in vitro combination therapy with polymyxin B on delaying resistance in Klebsiella pneumoniae and insights into the mechanisms of polymyxin-induced resistance
Objective: This study aimed to investigate the effects of polymyxin B (PMB) in combination with other antibiotics on delaying resistance in Klebsiella pneumoniae and to explore the mechanisms underlying PMB-induced resistance. Methods: In vitro continuous induction experiments were performed to obse...
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| Format: | Article |
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Elsevier
2025-06-01
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| Series: | Journal of Global Antimicrobial Resistance |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213716525000840 |
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| author | Huanhuan Zhang Wei Feng Fengjun Sun Te Xu Yan Qian |
| author_facet | Huanhuan Zhang Wei Feng Fengjun Sun Te Xu Yan Qian |
| author_sort | Huanhuan Zhang |
| collection | DOAJ |
| description | Objective: This study aimed to investigate the effects of polymyxin B (PMB) in combination with other antibiotics on delaying resistance in Klebsiella pneumoniae and to explore the mechanisms underlying PMB-induced resistance. Methods: In vitro continuous induction experiments were performed to observe changes in drug susceptibility with PMB alone vs. in combination. RNA-seq, quantitative reverse transcription PCR, and proteomic analyses were utilized to evaluate differential gene and protein expression between induced-resistant strains and those exhibiting delayed resistance. Then, gene knockout experiments were performed to validate the functional roles of relevant genes. Results: These findings indicated that PMB alone could induce resistance within 1–2 d, whereas the combination with amikacin (AMK) or tigecycline delayed the onset of resistance by 6 d. RNA-seq, quantitative reverse transcription PCR, and proteomic analyses revealed significant upregulation of nlpE, two-component systems, and AcrAB-TolC efflux pump-associated genes in PMB-induced resistant strains, whereas these genes were downregulated in the delayed resistant strains of PMB combined with AMK. Deletion and complementation experiments demonstrated that the expression levels of two-component systems and efflux pump-related genes were downregulated in nlpE knockout strains. Furthermore, PMB induction experiments revealed a significant upregulation of PmrA, PhoP, PhoQ, PagP, and AcrB proteins associated with cationic antimicrobial peptide pathways in the wild-type and nlpE complemented strains, whereas no differential change was observed in the nlpE knockout strain. Conclusions: nlpE contributes to PMB resistance by modulating the AcrAB-TolC efflux pump and the PhoP/Q and PmrA/B two-component systems. The combined use of PMB with AMK effectively delays the development of resistance in K. pneumoniae through the regulation of nlpE and its associated signalling pathways. |
| format | Article |
| id | doaj-art-2869a7d19b7442ab9ea3ee1a25594d64 |
| institution | Kabale University |
| issn | 2213-7165 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Global Antimicrobial Resistance |
| spelling | doaj-art-2869a7d19b7442ab9ea3ee1a25594d642025-08-20T03:24:13ZengElsevierJournal of Global Antimicrobial Resistance2213-71652025-06-014314315410.1016/j.jgar.2025.04.009Effects of in vitro combination therapy with polymyxin B on delaying resistance in Klebsiella pneumoniae and insights into the mechanisms of polymyxin-induced resistanceHuanhuan Zhang0Wei Feng1Fengjun Sun2Te Xu3Yan Qian4Department of Pharmacy, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China; Department of Pharmacy, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, ChinaDepartment of Pharmacy, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, ChinaDepartment of Pharmacy, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, ChinaDepartment of Pharmacy, The First People’s Hospital of Longquanyi District, Chengdu, ChinaDepartment of Pharmacy, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China; Corresponding author. Mailing address: Department of Pharmacy, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.Objective: This study aimed to investigate the effects of polymyxin B (PMB) in combination with other antibiotics on delaying resistance in Klebsiella pneumoniae and to explore the mechanisms underlying PMB-induced resistance. Methods: In vitro continuous induction experiments were performed to observe changes in drug susceptibility with PMB alone vs. in combination. RNA-seq, quantitative reverse transcription PCR, and proteomic analyses were utilized to evaluate differential gene and protein expression between induced-resistant strains and those exhibiting delayed resistance. Then, gene knockout experiments were performed to validate the functional roles of relevant genes. Results: These findings indicated that PMB alone could induce resistance within 1–2 d, whereas the combination with amikacin (AMK) or tigecycline delayed the onset of resistance by 6 d. RNA-seq, quantitative reverse transcription PCR, and proteomic analyses revealed significant upregulation of nlpE, two-component systems, and AcrAB-TolC efflux pump-associated genes in PMB-induced resistant strains, whereas these genes were downregulated in the delayed resistant strains of PMB combined with AMK. Deletion and complementation experiments demonstrated that the expression levels of two-component systems and efflux pump-related genes were downregulated in nlpE knockout strains. Furthermore, PMB induction experiments revealed a significant upregulation of PmrA, PhoP, PhoQ, PagP, and AcrB proteins associated with cationic antimicrobial peptide pathways in the wild-type and nlpE complemented strains, whereas no differential change was observed in the nlpE knockout strain. Conclusions: nlpE contributes to PMB resistance by modulating the AcrAB-TolC efflux pump and the PhoP/Q and PmrA/B two-component systems. The combined use of PMB with AMK effectively delays the development of resistance in K. pneumoniae through the regulation of nlpE and its associated signalling pathways.http://www.sciencedirect.com/science/article/pii/S2213716525000840Klebsiella pneumoniaePolymyxin BInduced resistance mechanismCombination therapyDelay resistance |
| spellingShingle | Huanhuan Zhang Wei Feng Fengjun Sun Te Xu Yan Qian Effects of in vitro combination therapy with polymyxin B on delaying resistance in Klebsiella pneumoniae and insights into the mechanisms of polymyxin-induced resistance Journal of Global Antimicrobial Resistance Klebsiella pneumoniae Polymyxin B Induced resistance mechanism Combination therapy Delay resistance |
| title | Effects of in vitro combination therapy with polymyxin B on delaying resistance in Klebsiella pneumoniae and insights into the mechanisms of polymyxin-induced resistance |
| title_full | Effects of in vitro combination therapy with polymyxin B on delaying resistance in Klebsiella pneumoniae and insights into the mechanisms of polymyxin-induced resistance |
| title_fullStr | Effects of in vitro combination therapy with polymyxin B on delaying resistance in Klebsiella pneumoniae and insights into the mechanisms of polymyxin-induced resistance |
| title_full_unstemmed | Effects of in vitro combination therapy with polymyxin B on delaying resistance in Klebsiella pneumoniae and insights into the mechanisms of polymyxin-induced resistance |
| title_short | Effects of in vitro combination therapy with polymyxin B on delaying resistance in Klebsiella pneumoniae and insights into the mechanisms of polymyxin-induced resistance |
| title_sort | effects of in vitro combination therapy with polymyxin b on delaying resistance in klebsiella pneumoniae and insights into the mechanisms of polymyxin induced resistance |
| topic | Klebsiella pneumoniae Polymyxin B Induced resistance mechanism Combination therapy Delay resistance |
| url | http://www.sciencedirect.com/science/article/pii/S2213716525000840 |
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