Abstract
Developing an effective therapy to overcome carbapenemase-positive Klebsiella pneumoniae (CPKp) is an important therapeutic challenge that must be addressed urgently. Here, we explored a Ca-EDTA combination with aztreonam or ceftazidime-avibactam in vitro and in vivo against diverse CPKp clinical isolates. The synergy testing of this study demonstrated that novel aztreonam-Ca-EDTA or ceftazidime-avibactam-Ca-EDTA combination was significantly effective in eliminating planktonic and mature biofilms in vitro, as well as eradicating CPKp infections in vivo. Both combinations revealed significant therapeutic efficacies in reducing bacterial load in internal organs and protecting treated mice from mortality. Conclusively, this is the first in vitro and in vivo study to demonstrate that novel aztreonam-Ca-EDTA or ceftazidime-avibactam-Ca-EDTA combinations provide favorable efficacy and safety for successful eradication of carbapenemase-producing Klebsiella pneumoniae planktonic and biofilm infections.
| Original language | English |
|---|---|
| Article number | 107215 |
| Pages (from-to) | 1-24 |
| Number of pages | 24 |
| Journal | iScience |
| Volume | 26 |
| Issue number | 7 |
| DOIs | |
| Publication status | Published - 21 Jul 2023 |
Bibliographical note
Funding Information:We thank the staff of the bacteriology division, Department of Microbiology at King Chulalongkorn Memorial Hospital, for providing the K. pneumoniae clinical isolates. Funding: This work was supported by a grant from the 90th Year Anniversary Ratchadapiseksompotch Endowment Fund from the Faculty of Medicine and Graduate School, Chulalongkorn University, Bangkok, Thailand (batch No. 39 (2/61)). This research is funded by Thailand Science Research and Innovation Fund Chulalongkorn University (CU_FRB65_hea(43)_050_30_31). 90th Year Anniversary Ratchadapiseksompotch Endowment Fund from the Graduate School, Chulalongkorn University, Bangkok, Thailand supported Sasipen Sae-Joo. Aye Mya Sithu Shein was supported by Chulalonslgkorn University (Second Century Fund- C2F Fellowship). Dhammika Leshan Wannigama was supported by Chulalongkorn University (Second Century Fund- C2F Fellowship), the University of Western Australia (Overseas Research Experience Fellowship), and Yamagata Prefectural Central Hospital, Yamagata, Japan (Clinical Residency Fellowship). SMS holds an NHMRC Investigator Grant (2007725) and AK is a Rothwell Family Fellow. The sponsor(s) had no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the article for publication. The study protocol was approved by the Institutional Review Board (IRB) of the Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand (Certificate of approval (COA) No. 045/2020, IRB No. 774/63), and experiments were conducted in compliance with national and international ethical guidelines for human research as specified in the Declaration of Helsinki (1964) and its contemporary (2013) amendments, and comparable ethical standards including The Belmont Report, Council for International Organizations of Medical Sciences (CIOMS) Guidelines, and International Conference on Harmonization in Good Clinical Practice (ICH-GCP). For this retrospective study of anonymized clinical isolates, the requirement for informed consent from patients was waived by the IRB. All protocols involving animals (C57BL/6 mice) conformed to the revised guidelines of the U.S. Institute of Laboratory Animal Resources, Commission on Life Sciences, National Research Council Guide for the Care and Use of Laboratory Animals, Washington, DC: National Academy Press; 1996, and the Animals for Scientific Purposes Act, 2015 (BE 2558), and experiments were performed following the ethical standards as laid down in the Basel Declaration on the use of animals in research. The Institutional Animal Care and Use Committee (IACUC) of the Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand, approved the protocol (Certificate No. 033/2563, Research Project No. 020/2563) and protocols performed by operators licensed by the Thai Institute for Animals for Scientific Purpose Development and National Research Council of Thailand. The present article was prepared in compliance with the ARRIVE 2.0 guidelines for reporting animal research. For this retrospective study of anonymous clinical isolates, the requirement for informed consent from patients was waived by the Institutional Review Board (IRB) of the Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand (COA No. 045/2020, IRB No. 774/63). D.L.W.: Conception, investigation, funding acquisition, data curation, formal analysis, and writing the original draft of the manuscript. A.M.S.S.: Data curation, formal analysis, editing the original draft of the manuscript, and contributed equally to this work as first authors. C.H.: Formal analysis, supervision, methodology, validation, critical review, editing of the manuscript, and contributed equally to this work as first authors. P.N.M.: Supervision, critical review, and editing of the manuscript. P.H.: Formal analysis, supervision, methodology, validation, critical review, editing of the manuscript, and contributed equally to this work as first authors. P.P.: Critical review and editing of the manuscript. W.G.F.D.: Critical review and editing of the manuscript. P.O.: Critical review and editing of the manuscript. T.S.: Bioinformatics analysis, methodology, and validation. N.C.: Confocal laser scanning microscopy data acquisition, curation, methodology, and validation. P.W.: Bioinformatics analysis, methodology, and validation. S.L.: In-data acquisition, curation, formal analysis, methodology, validation, critical review, and editing of the manuscript. S.S-J.: Bacteria identification and clinical collection. S.N.: Bacteria identification and clinical collection. U.R.: Bacteria identification and clinical collection. C.T.: Data acquisition and curation. N.K.: Bacteria identification and clinical collection. M.L.: Bacteria identification and clinical collection. T.L.: Conception for mouse model, formal analysis. R.K.: Conception for mouse model, formal analysis, supervision, critical review and editing of the manuscript. R.R.: Ca-EDTA preparation and critical review, editing of the manuscript. N.N.: Critical review, editing of the manuscript. S.T.: Critical review, editing of the manuscript. A.L.: Critical review and editing of the manuscript. N.K.D.R.: Critical review, editing of the manuscript. V.N.B.: Ca-EDTA preparation and critical review, editing of the manuscript. S.M.A.H.R.: Critical review, editing of the manuscript. T.K.: Critical review and editing of the manuscript. N.H.: Critical review and editing of the manuscript. R.J.S.: Critical review and editing of the manuscript. LC.: Critical review, editing of the manuscript. M.A.: Supervision, critical review and editing of the manuscript. H.I.: Critical review, editing of the manuscript. P.G.H.: Supervision, critical review and editing of the manuscript and contributed equally to this work as first authors. A.K.: Supervision, critical review and editing of the manuscript. S.MS: Supervision, critical review and editing of the manuscript. T.C.: Supervision, critical review and editing of the manuscript. S.A.: Supervision, conception, investigation, funding acquisition, data curation, critical review and editing of the manuscript. The authors declare no competing interests. We support inclusive, diverse, and equitable conduct of research. One or more of the authors of this paper self-identifies as an underrepresented ethnic minority in their field of research or within their geographical location. One or more of the authors of this paper self-identifies as a gender minority in their field of research. One or more of the authors of this paper self-identifies as a member of the LGBTQIA+ community. One or more of the authors of this paper self-identifies as living with a disability. While citing references scientifically relevant for this work, we also actively worked to promote gender balance in our reference list.
Funding Information:
All protocols involving animals (C57BL/6 mice) conformed to the revised guidelines of the U.S. Institute of Laboratory Animal Resources, Commission on Life Sciences, National Research Council Guide for the Care and Use of Laboratory Animals, Washington, DC: National Academy Press; 1996, and the Animals for Scientific Purposes Act, 2015 (BE 2558), and experiments were performed following the ethical standards as laid down in the Basel Declaration on the use of animals in research. The Institutional Animal Care and Use Committee (IACUC) of the Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand, approved the protocol (Certificate No. 033/2563, Research Project No. 020/2563) and protocols performed by operators licensed by the Thai Institute for Animals for Scientific Purpose Development and National Research Council of Thailand. The present article was prepared in compliance with the ARRIVE 2.0 guidelines for reporting animal research.
Funding Information:
Funding: This work was supported by a grant from the 90th Year Anniversary Ratchadapiseksompotch Endowment Fund from the Faculty of Medicine and Graduate School, Chulalongkorn University , Bangkok, Thailand (batch No. 39 (2/61)). This research is funded by Thailand Science Research and Innovation Fund Chulalongkorn University ( CU_FRB65_hea(43)_050_30_31 ). 90th Year Anniversary Ratchadapiseksompotch Endowment Fund from the Graduate School, Chulalongkorn University, Bangkok, Thailand supported Sasipen Sae-Joo. Aye Mya Sithu Shein was supported by Chulalonslgkorn University (Second Century Fund- C2F Fellowship). Dhammika Leshan Wannigama was supported by Chulalongkorn University (Second Century Fund- C2F Fellowship), the University of Western Australia (Overseas Research Experience Fellowship), and Yamagata Prefectural Central Hospital , Yamagata, Japan (Clinical Residency Fellowship). SMS holds an NHMRC Investigator Grant ( 2007725 ) and AK is a Rothwell Family Fellow. The sponsor(s) had no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the article for publication.
Publisher Copyright:
© 2023 The Author(s)
