title: Inhibition of the β-Lactamase BlaMabby Avibactam Improves theIn VitroandIn VivoEfficacy of Imipenem against Mycobacterium abscessus.
abstract: Mycobacterium abscessuspulmonary infections are treated with a macrolide (clarithromycin or azithromycin), an aminoglycoside (amikacin), and a β-lactam (cefoxitin or imipenem). The triple combination is used without any β-lactamase inhibitor, even thoughMabscessusproduces the broad-spectrum β-lactamase BlaMabWe determine whether inhibition of BlaMabby avibactam improves the activity of imipenem againstM. abscessusThe bactericidal activity of drug combinations was assayed in broth and in human macrophages. Thein vivoefficacy of the drugs was tested by monitoring the survival of infected zebrafish embryos. The level of BlaMabproduction in broth and in macrophages was compared by quantitative reverse transcription-PCR and Western blotting. The triple combination of imipenem (8 or 32 μg/ml), amikacin (32 μg/ml), and avibactam (4 μg/ml) was bactericidal in broth (<0.1% survival), with 3.2- and 4.3-log10reductions in the number of CFU being achieved at 72 h when imipenem was used at 8 and 32 μg/ml, respectively. The triple combination achieved significant intracellular killing, with the bacterial survival rates being 54% and 7% with the low (8 μg/ml) and high (32 μg/ml) dosages of imipenem, respectively.In vivoinhibition of BlaMabby avibactam improved the survival of zebrafish embryos treated with imipenem. Expression of the gene encoding BlaMabwas induced (20-fold) in the infected macrophages. Inhibition of BlaMabby avibactam improved the efficacy of imipenem againstM. abscessusin vitro, in macrophages, and in zebrafish embryos, indicating that this β-lactamase inhibitor should be clinically evaluated. Thein vitroevaluation of imipenem may underestimate the impact of BlaMab, since the production of the β-lactamase is inducible in macrophages.
authors.affiliation: INSERM, U1138, LRMA, Equipe 12 du Centre de Recherche des Cordeliers, Paris, France.|Université Pierre et Marie Curie, UMR S 1138, Paris, France.|Université Paris Descartes, Sorbonne Paris Cité, UMR S 1138, Paris, France.
authors.forename: Anne-Laure
authors.initials: AL
authors.lastname: Lefebvre
authors.affiliation: UMR1173, INSERM, Université de Versailles St. Quentin, Montigny le Bretonneux, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Le Moigne
authors.affiliation: Centre d'Études d'Agents Pathogènes et Biotechnologies pour la Santé, FR3689, CNRS, Université Montpellier, Montpellier, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Bernut
authors.affiliation: INSERM, U1138, LRMA, Equipe 12 du Centre de Recherche des Cordeliers, Paris, France.|Université Pierre et Marie Curie, UMR S 1138, Paris, France.|Université Paris Descartes, Sorbonne Paris Cité, UMR S 1138, Paris, France.
authors.forename: Carole
authors.initials: C
authors.lastname: Veckerlé
authors.affiliation: INSERM, U1138, LRMA, Equipe 12 du Centre de Recherche des Cordeliers, Paris, France.|Université Pierre et Marie Curie, UMR S 1138, Paris, France.|Université Paris Descartes, Sorbonne Paris Cité, UMR S 1138, Paris, France.|Assistance Publique-Hôpitaux de Paris, Service de Microbiologie, Hôpital Européen Georges Pompidou, Paris, France.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Compain
authors.affiliation: UMR1173, INSERM, Université de Versailles St. Quentin, Montigny le Bretonneux, France.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Herrmann
authors.affiliation: Centre d'Études d'Agents Pathogènes et Biotechnologies pour la Santé, FR3689, CNRS, Université Montpellier, Montpellier, France.|INSERM, CPBS, Montpellier, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Kremer
authors.affiliation: INSERM, U1138, LRMA, Equipe 12 du Centre de Recherche des Cordeliers, Paris, France michel.arthur@crc.jussieu.fr jean-luc.mainardi@crc.jussieu.fr.|Université Pierre et Marie Curie, UMR S 1138, Paris, France.|Université Paris Descartes, Sorbonne Paris Cité, UMR S 1138, Paris, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Arthur
authors.affiliation: INSERM, U1138, LRMA, Equipe 12 du Centre de Recherche des Cordeliers, Paris, France michel.arthur@crc.jussieu.fr jean-luc.mainardi@crc.jussieu.fr.|Université Pierre et Marie Curie, UMR S 1138, Paris, France.|Université Paris Descartes, Sorbonne Paris Cité, UMR S 1138, Paris, France.|Assistance Publique-Hôpitaux de Paris, Service de Microbiologie, Hôpital Européen Georges Pompidou, Paris, France.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Mainardi
mesh_terms: D000583:Amikacin / Q000494:pharmacology; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D053961:Azabicyclo Compounds / Q000494:pharmacology*; D015153:Blotting, Western; D004625:Embryo, Nonmammalian / Q000382:microbiology; D006801:Humans; D015378:Imipenem / Q000494:pharmacology*; D008264:Macrophages / Q000382:microbiology; D008826:Microbial Sensitivity Tests; D009161:Mycobacterium / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015027:Zebrafish; D065093:beta-Lactamase Inhibitors / Q000494:pharmacology*; D001618:beta-Lactamases / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D053961:Azabicyclo Compounds; D065093:beta-Lactamase Inhibitors; D015378:Imipenem; C543519:avibactam; D000583:Amikacin; D001618:beta-Lactamases
keywords: Mycobacterium abscessus; avibactam; cystic fibrosis; imipenem; β-lactamase inhibitor