Diverse evolutionary trajectories of Klebsiella pneumoniae carbapenemase: unraveling the impact of amino acid substitutions on β-lactam susceptibility and the role of avibactam in driving resistance
ABSTRACT Klebsiella pneumoniae carbapenemases (KPCs) have evolved into over 245 distinct variants, with over one-third of variants exhibiting reduced susceptibility to ceftazidime-avibactam, while the underlying selection mechanisms remain elusive. To better elucidate these resistant phenotypes, we...
Saved in:
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2025-04-01
|
| Series: | mSystems |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/msystems.00184-25 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850156450485108736 |
|---|---|
| author | Jie Wei Jinyu Huang Chunhong Zou Shimei Shen Barry N. Kreiswirth Ailong Huang Shifeng Huang Liang Chen Deqiang Wang Siqiang Niu |
| author_facet | Jie Wei Jinyu Huang Chunhong Zou Shimei Shen Barry N. Kreiswirth Ailong Huang Shifeng Huang Liang Chen Deqiang Wang Siqiang Niu |
| author_sort | Jie Wei |
| collection | DOAJ |
| description | ABSTRACT Klebsiella pneumoniae carbapenemases (KPCs) have evolved into over 245 distinct variants, with over one-third of variants exhibiting reduced susceptibility to ceftazidime-avibactam, while the underlying selection mechanisms remain elusive. To better elucidate these resistant phenotypes, we cloned 33 clinically described KPC variants (from KPC-2 to KPC-36) and 8 artificially created variants into a common plasmid vector and assessed their impact on β-lactam susceptibility. Strains expressing KPC-14, KPC-28, and KPC-31 exhibited increased resistance to ceftazidime and ceftazidime-avibactam but decreased resistance to carbapenems. We further studied the catalytic mechanism of β-lactam hydrolysis by KPC-4, KPC-14, KPC-15, KPC-16, KPC-21, KPC-25, KPC-28, KPC-31, and the ancestral KPC-2 and KPC-3 enzymes. Antimicrobial susceptibility test, enzyme kinetics, and molecular modeling revealed diverse selective pressures, including but not limited to aztreonam and ceftriaxone, driving KPC evolution, with ceftazidime playing a central role. Substitutions within the KPC hydrolytic active sites notably reduced the inhibitory effect of avibactam on KPC, demonstrated by isothermal titration calorimetry analysis, resulting in enhanced hydrolysis of ceftazidime by enzyme kinetics. This highlights that avibactam may serve as an additional driving force in KPC evolution.IMPORTANCEThe rapid evolution of KPC carbapenemases, including resistance to ceftazidime-avibactam, threatens the effectiveness of last-resort antibiotics against Klebsiella pneumoniae infections, necessitating understanding of of the underlying selection pressures. This study investigates the evolutionary mechanisms driving KPC diversification and resistance to ceftazidime-avibactam, providing crucial information for developing effective strategies to combat carbapenem-resistant Klebsiella pneumoniae (CRKP) infections and preserve antibiotic efficacy. |
| format | Article |
| id | doaj-art-d84998a2bd8a4a51a05c08d9d0000019 |
| institution | OA Journals |
| issn | 2379-5077 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSystems |
| spelling | doaj-art-d84998a2bd8a4a51a05c08d9d00000192025-08-20T02:24:33ZengAmerican Society for MicrobiologymSystems2379-50772025-04-0110410.1128/msystems.00184-25Diverse evolutionary trajectories of Klebsiella pneumoniae carbapenemase: unraveling the impact of amino acid substitutions on β-lactam susceptibility and the role of avibactam in driving resistanceJie Wei0Jinyu Huang1Chunhong Zou2Shimei Shen3Barry N. Kreiswirth4Ailong Huang5Shifeng Huang6Liang Chen7Deqiang Wang8Siqiang Niu9Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Yuzhong, Chongqing, ChinaCollege of Laboratory Medicine, Chongqing Medical University, Yuzhong, Chongqing, ChinaDepartment of Clinical Laboratory, University-Town Hospital of Chongqing Medical University, Chongqing, ChinaCollege of Laboratory Medicine, Chongqing Medical University, Yuzhong, Chongqing, ChinaCenter for Discovery and Innovation Hackensack Meridian Health, Nutley, New Jersey, USAThe Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education Chongqing Medical University, Yuzhong, Chongqing, ChinaDepartment of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Yuzhong, Chongqing, ChinaDepartment of Pharmacy Practice, School of Pharmacy and Pharmaceutical Sciences University at Buffalo, Buffalo, New York, USAThe Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education Chongqing Medical University, Yuzhong, Chongqing, ChinaDepartment of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Yuzhong, Chongqing, ChinaABSTRACT Klebsiella pneumoniae carbapenemases (KPCs) have evolved into over 245 distinct variants, with over one-third of variants exhibiting reduced susceptibility to ceftazidime-avibactam, while the underlying selection mechanisms remain elusive. To better elucidate these resistant phenotypes, we cloned 33 clinically described KPC variants (from KPC-2 to KPC-36) and 8 artificially created variants into a common plasmid vector and assessed their impact on β-lactam susceptibility. Strains expressing KPC-14, KPC-28, and KPC-31 exhibited increased resistance to ceftazidime and ceftazidime-avibactam but decreased resistance to carbapenems. We further studied the catalytic mechanism of β-lactam hydrolysis by KPC-4, KPC-14, KPC-15, KPC-16, KPC-21, KPC-25, KPC-28, KPC-31, and the ancestral KPC-2 and KPC-3 enzymes. Antimicrobial susceptibility test, enzyme kinetics, and molecular modeling revealed diverse selective pressures, including but not limited to aztreonam and ceftriaxone, driving KPC evolution, with ceftazidime playing a central role. Substitutions within the KPC hydrolytic active sites notably reduced the inhibitory effect of avibactam on KPC, demonstrated by isothermal titration calorimetry analysis, resulting in enhanced hydrolysis of ceftazidime by enzyme kinetics. This highlights that avibactam may serve as an additional driving force in KPC evolution.IMPORTANCEThe rapid evolution of KPC carbapenemases, including resistance to ceftazidime-avibactam, threatens the effectiveness of last-resort antibiotics against Klebsiella pneumoniae infections, necessitating understanding of of the underlying selection pressures. This study investigates the evolutionary mechanisms driving KPC diversification and resistance to ceftazidime-avibactam, providing crucial information for developing effective strategies to combat carbapenem-resistant Klebsiella pneumoniae (CRKP) infections and preserve antibiotic efficacy.https://journals.asm.org/doi/10.1128/msystems.00184-25KPC (Klebsiella pneumoniae carbapenemase)phylogenetic reconstructionceftazidime-avibactamevolutionresistance |
| spellingShingle | Jie Wei Jinyu Huang Chunhong Zou Shimei Shen Barry N. Kreiswirth Ailong Huang Shifeng Huang Liang Chen Deqiang Wang Siqiang Niu Diverse evolutionary trajectories of Klebsiella pneumoniae carbapenemase: unraveling the impact of amino acid substitutions on β-lactam susceptibility and the role of avibactam in driving resistance mSystems KPC (Klebsiella pneumoniae carbapenemase) phylogenetic reconstruction ceftazidime-avibactam evolution resistance |
| title | Diverse evolutionary trajectories of Klebsiella pneumoniae carbapenemase: unraveling the impact of amino acid substitutions on β-lactam susceptibility and the role of avibactam in driving resistance |
| title_full | Diverse evolutionary trajectories of Klebsiella pneumoniae carbapenemase: unraveling the impact of amino acid substitutions on β-lactam susceptibility and the role of avibactam in driving resistance |
| title_fullStr | Diverse evolutionary trajectories of Klebsiella pneumoniae carbapenemase: unraveling the impact of amino acid substitutions on β-lactam susceptibility and the role of avibactam in driving resistance |
| title_full_unstemmed | Diverse evolutionary trajectories of Klebsiella pneumoniae carbapenemase: unraveling the impact of amino acid substitutions on β-lactam susceptibility and the role of avibactam in driving resistance |
| title_short | Diverse evolutionary trajectories of Klebsiella pneumoniae carbapenemase: unraveling the impact of amino acid substitutions on β-lactam susceptibility and the role of avibactam in driving resistance |
| title_sort | diverse evolutionary trajectories of klebsiella pneumoniae carbapenemase unraveling the impact of amino acid substitutions on β lactam susceptibility and the role of avibactam in driving resistance |
| topic | KPC (Klebsiella pneumoniae carbapenemase) phylogenetic reconstruction ceftazidime-avibactam evolution resistance |
| url | https://journals.asm.org/doi/10.1128/msystems.00184-25 |
| work_keys_str_mv | AT jiewei diverseevolutionarytrajectoriesofklebsiellapneumoniaecarbapenemaseunravelingtheimpactofaminoacidsubstitutionsonblactamsusceptibilityandtheroleofavibactamindrivingresistance AT jinyuhuang diverseevolutionarytrajectoriesofklebsiellapneumoniaecarbapenemaseunravelingtheimpactofaminoacidsubstitutionsonblactamsusceptibilityandtheroleofavibactamindrivingresistance AT chunhongzou diverseevolutionarytrajectoriesofklebsiellapneumoniaecarbapenemaseunravelingtheimpactofaminoacidsubstitutionsonblactamsusceptibilityandtheroleofavibactamindrivingresistance AT shimeishen diverseevolutionarytrajectoriesofklebsiellapneumoniaecarbapenemaseunravelingtheimpactofaminoacidsubstitutionsonblactamsusceptibilityandtheroleofavibactamindrivingresistance AT barrynkreiswirth diverseevolutionarytrajectoriesofklebsiellapneumoniaecarbapenemaseunravelingtheimpactofaminoacidsubstitutionsonblactamsusceptibilityandtheroleofavibactamindrivingresistance AT ailonghuang diverseevolutionarytrajectoriesofklebsiellapneumoniaecarbapenemaseunravelingtheimpactofaminoacidsubstitutionsonblactamsusceptibilityandtheroleofavibactamindrivingresistance AT shifenghuang diverseevolutionarytrajectoriesofklebsiellapneumoniaecarbapenemaseunravelingtheimpactofaminoacidsubstitutionsonblactamsusceptibilityandtheroleofavibactamindrivingresistance AT liangchen diverseevolutionarytrajectoriesofklebsiellapneumoniaecarbapenemaseunravelingtheimpactofaminoacidsubstitutionsonblactamsusceptibilityandtheroleofavibactamindrivingresistance AT deqiangwang diverseevolutionarytrajectoriesofklebsiellapneumoniaecarbapenemaseunravelingtheimpactofaminoacidsubstitutionsonblactamsusceptibilityandtheroleofavibactamindrivingresistance AT siqiangniu diverseevolutionarytrajectoriesofklebsiellapneumoniaecarbapenemaseunravelingtheimpactofaminoacidsubstitutionsonblactamsusceptibilityandtheroleofavibactamindrivingresistance |