High-level ceftazidime-avibactam resistance by in-host evolution of blaKPC genes: Emergence of a novel blaKPC-102 variant and increase of blaKPC-33 copy number in Klebsiella pneumoniae strains from a lung transplantation recipient

IF 4.9 2区医学 Q1 INFECTIOUS DISEASES

International Journal of Antimicrobial Agents Pub Date : 2025-04-21 DOI:10.1016/j.ijantimicag.2025.107521

Yulin Zhang , Ziyao Li , Huihui Shi , Zichen Lei , Dongya Pu , Qi Liu , Feilong Zhang , Jiankang Zhao , Xinmeng Liu , Binghuai Lu , Bin Cao

{"title":"High-level ceftazidime-avibactam resistance by in-host evolution of blaKPC genes: Emergence of a novel blaKPC-102 variant and increase of blaKPC-33 copy number in Klebsiella pneumoniae strains from a lung transplantation recipient","authors":"Yulin Zhang , Ziyao Li , Huihui Shi , Zichen Lei , Dongya Pu , Qi Liu , Feilong Zhang , Jiankang Zhao , Xinmeng Liu , Binghuai Lu , Bin Cao","doi":"10.1016/j.ijantimicag.2025.107521","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><div>KPC-producing <em>Klebsiella pneumoniae</em> (Kp) infections have become one of the major threats to public health in China. Recently, the emergence of ceftazidime-avibactam (CZA) resistance due to the <em>bla</em><sub>KPC</sub> mutations has been increasingly reported.</div></div><div><h3>Methods</h3><div>Two CZA-susceptible Kp strains (Kp36854 carrying <em>bla</em><sub>KPC-2</sub> and Kp37523 without <em>bla</em><sub>KPC</sub>) and two CZA-resistant Kp strains (Kp38935 carrying double copies of <em>bla</em><sub>KPC-33</sub> and Kp38097 carrying a newly identified <em>bla</em><sub>KPC-102</sub> gene) were isolated from a lung-transplanted patient during CZA treatment. This study analyzed the within-host evolutionary dynamics of <em>bla</em><sub>KPC</sub> in KPC-Kp strains.</div></div><div><h3>Results</h3><div>Compared with KPC-2, KPC-33 possessed only a D179Y substitution while KPC-102 harbored both D179Y and Y241D substitutions. We constructed KPC-δ33 with only a Y241D substitution, which means the correction of the first mutation that is also present in KPC-33 (D179Y). Cloning and expression experiments showed that CZA-MIC value of <em>E. coli</em> DH5α/pKPC102 was 512 mg/L, while CZA-MIC values of both <em>E. coli</em> DH5α/pKPC-33 and <em>E. coli</em> DH5α/pKPC-δ33 were 32/4 mg/L. Enzymatic kinetic analysis revealed that KPC-2 demonstrated the highest catalytic efficiency to nitrocefin and meropenem, whereas KPC-33 displayed higher catalytic efficiency against ceftazidime compared to other tested KPC variants. Regarding avibactam, KPC-2 showed the highest sensitivity, while KPC-δ33 and KPC-102 were less sensitive.</div></div><div><h3>Conclusion</h3><div>High-level CZA resistance was mediated by the novel <em>bla</em><sub>KPC-102</sub> and double copies of <em>bla</em><sub>KPC-33</sub> gene, respectively, in two porin-deficient Kp strains from one patient. The CZA resistance resulting from <em>bla</em><sub>KPC</sub> mutation could be selected and evolved to be more diverse and heterogeneous within the host after CZA therapy. It is very essential to perform the surveillance of CZA-resistance for clinicians during treatment.</div></div>","PeriodicalId":13818,"journal":{"name":"International Journal of Antimicrobial Agents","volume":"66 2","pages":"Article 107521"},"PeriodicalIF":4.9000,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"International Journal of Antimicrobial Agents","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0924857925000780","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"INFECTIOUS DISEASES","Score":null,"Total":0}

引用次数: 0

Abstract

Objective

KPC-producing Klebsiella pneumoniae (Kp) infections have become one of the major threats to public health in China. Recently, the emergence of ceftazidime-avibactam (CZA) resistance due to the bla_KPC mutations has been increasingly reported.

Methods

Two CZA-susceptible Kp strains (Kp36854 carrying bla_KPC-2 and Kp37523 without bla_KPC) and two CZA-resistant Kp strains (Kp38935 carrying double copies of bla_KPC-33 and Kp38097 carrying a newly identified bla_KPC-102 gene) were isolated from a lung-transplanted patient during CZA treatment. This study analyzed the within-host evolutionary dynamics of bla_KPC in KPC-Kp strains.

Results

Compared with KPC-2, KPC-33 possessed only a D179Y substitution while KPC-102 harbored both D179Y and Y241D substitutions. We constructed KPC-δ33 with only a Y241D substitution, which means the correction of the first mutation that is also present in KPC-33 (D179Y). Cloning and expression experiments showed that CZA-MIC value of E. coli DH5α/pKPC102 was 512 mg/L, while CZA-MIC values of both E. coli DH5α/pKPC-33 and E. coli DH5α/pKPC-δ33 were 32/4 mg/L. Enzymatic kinetic analysis revealed that KPC-2 demonstrated the highest catalytic efficiency to nitrocefin and meropenem, whereas KPC-33 displayed higher catalytic efficiency against ceftazidime compared to other tested KPC variants. Regarding avibactam, KPC-2 showed the highest sensitivity, while KPC-δ33 and KPC-102 were less sensitive.

Conclusion

High-level CZA resistance was mediated by the novel bla_KPC-102 and double copies of bla_KPC-33 gene, respectively, in two porin-deficient Kp strains from one patient. The CZA resistance resulting from bla_KPC mutation could be selected and evolved to be more diverse and heterogeneous within the host after CZA therapy. It is very essential to perform the surveillance of CZA-resistance for clinicians during treatment.

Abstract Image

查看原文本刊更多论文

blaKPC基因在宿主内进化对头孢他啶-阿维巴坦的高水平耐药性：来自肺移植受体的肺炎克雷伯菌菌株出现新的blaKPC-102变体和blaKPC-33拷贝数的增加

目的产kpc肺炎克雷伯菌（Kp）感染已成为中国公共卫生的主要威胁之一。最近，由于blaKPC突变引起的头孢他啶-阿维巴坦（CZA）耐药的报道越来越多。方法从肺移植患者中分离2株CZA敏感Kp菌株（Kp36854携带blaKPC-2和Kp37523不携带blaKPC）和2株CZA耐药Kp菌株（Kp38935携带blaKPC-33双拷贝和Kp38097携带新鉴定的blaKPC-102基因）。本研究分析了KPC-Kp菌株blaKPC在宿主内的进化动态。结果与KPC-2相比，KPC-33只具有一个D179Y取代，而KPC-102同时具有D179Y和Y241D取代。我们构建的KPC-δ33只有一个Y241D替换，这意味着修正了KPC-33 （D179Y）中也存在的第一个突变。克隆表达实验表明，大肠杆菌DH5α/pKPC102的CZA-MIC值为512 mg/L，大肠杆菌DH5α/pKPC-33和大肠杆菌DH5α/pKPC-δ33的CZA-MIC值均为32/4 mg/L。酶动力学分析表明，KPC-2对硝基萘和美罗培南的催化效率最高，而KPC-33对头孢他啶的催化效率高于其他KPC变体。对于阿维巴坦，KPC-2的敏感性最高，KPC-δ33和KPC-102的敏感性较低。结论来自同一患者的两株孔蛋白缺陷Kp菌株分别由新型blaKPC-102和双拷贝blaKPC-33基因介导了CZA的高水平耐药。经CZA治疗后，由blaKPC突变引起的CZA耐药在宿主体内可以被选择和进化为更加多样化和异质性。临床医生在治疗期间进行cza耐药性监测是非常必要的。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

International Journal of Antimicrobial Agents 医学-传染病学

CiteScore

21.60

自引率

0.90%

发文量

176

审稿时长

36 days

期刊介绍： The International Journal of Antimicrobial Agents is a peer-reviewed publication offering comprehensive and current reference information on the physical, pharmacological, in vitro, and clinical properties of individual antimicrobial agents, covering antiviral, antiparasitic, antibacterial, and antifungal agents. The journal not only communicates new trends and developments through authoritative review articles but also addresses the critical issue of antimicrobial resistance, both in hospital and community settings. Published content includes solicited reviews by leading experts and high-quality original research papers in the specified fields.