Combination of Gallium Citrate and Levofloxacin Induces a Distinct Metabolome Profile and Enhances Growth Inhibition of Multidrug-resistant Compared to Linezolid

Overview

Journal Front Microbiol

Specialty Microbiology

Date 2024 Dec 19

PMID 39697659

Authors

Oleksandr Ilchenko

Elena Nikolaevskaya

Oksana Zinchenko

Volodymyr Ivanytsia

Cristina Prat-Aymerich

Madeleine Ramstedt

Olena Rzhepishevska

Affiliations

Soon will be listed here.

Abstract

Introduction: Tuberculosis (TB) treatment typically involves a tailored combination of four antibiotics based on the drug resistance profile of the infecting strain. The increasing drug resistance of () requires the development of novel antibiotics to ensure effective treatment regimens. Gallium (Ga) is being explored as a repurposed drug against TB due to its ability to inhibit growth and disrupt iron metabolism. Given the potential interactions between Ga and established antibiotics, we investigated how a combination of Ga with levofloxacin (Lfx) or linezolid (Lzd) affects the growth and metabolome of a multidrug-resistant (MDR) clinical strain.

Methods: was cultured using a BACTEC 960 system with concentrations of Ga ranging from 125 to 1,000 μM and with 250 to 500 μM of Ga combined with 0.125 mg/L of Lfx or Lzd. For metabolome analysis, the antibacterials were used at concentrations that inhibited the growth of bacteria without causing cell death. Metabolites were extracted from cells and analyzed using chromatography-mass spectrometry.

Results: The MDR strain exhibited a dose-dependent response to Ga. Notably, the enhancement in growth inhibition was statistically significant for the Ga/Lfx combination compared to Ga alone, while no such significance was observed for Ga/Lzd. Moreover, exposure to Ga/Lfx or Ga/Lzd resulted in distinct metabolite profiles. Ga treatment increased the level of aconitate, fumarate, and glucose in the cells, suggesting the inhibition of iron-dependent aconitase and fumarate hydratase, as well as disruption of the pentose phosphate pathway. The levels of glucose, succinic acid, citric acid, and hexadecanoic acid followed a similar pattern in cells exposed to Ga and Ga/Lfx at 500 μM Ga but exhibited different trends at 250 μM Ga.

Discussion: In the presence of Lfx, the metabolome changes induced by Ga are more pronounced compared to those observed with Lzd. Lfx affects nucleic acids and transcription, which may enhance Ga-dependent growth inhibition by preventing the metabolic redirection that bacteria typically use to bypass iron-dependent enzymes.

References

Ruecker N, Jansen R, Trujillo C, Puckett S, Jayachandran P, Piroli G . Fumarase Deficiency Causes Protein and Metabolite Succination and Intoxicates Mycobacterium tuberculosis. Cell Chem Biol. 2017; 24(3):306-315. PMC: 5357164. DOI: 10.1016/j.chembiol.2017.01.005. View

Mor N, Vanderkolk J, Heifets L . Inhibitory and bactericidal activities of levofloxacin against Mycobacterium tuberculosis in vitro and in human macrophages. Antimicrob Agents Chemother. 1994; 38(5):1161-4. PMC: 188169. DOI: 10.1128/AAC.38.5.1161. View

Yurekten O, Payne T, Tejera N, Amaladoss F, Martin C, Williams M . MetaboLights: open data repository for metabolomics. Nucleic Acids Res. 2023; 52(D1):D640-D646. PMC: 10767962. DOI: 10.1093/nar/gkad1045. View

Reithuber E, Nannapaneni P, Rzhepishevska O, Lindgren A, Ilchenko O, Normark S . The Bactericidal Fatty Acid Mimetic 2CCA-1 Selectively Targets Pneumococcal Extracellular Polyunsaturated Fatty Acid Metabolism. mBio. 2020; 11(6). PMC: 7773995. DOI: 10.1128/mBio.03027-20. View

Lobritz M, Belenky P, Porter C, Gutierrez A, Yang J, Schwarz E . Antibiotic efficacy is linked to bacterial cellular respiration. Proc Natl Acad Sci U S A. 2015; 112(27):8173-80. PMC: 4500273. DOI: 10.1073/pnas.1509743112. View

Leitao R, Silva F, Ribeiro G, Santos I, Guerreiro J, Mendes F . Gallium and indium complexes with isoniazid-derived ligands: Interaction with biomolecules and biological activity against cancer cells and Mycobacterium tuberculosis. J Inorg Biochem. 2022; 240:112091. DOI: 10.1016/j.jinorgbio.2022.112091. View

Olakanmi O, Kesavalu B, Pasula R, Abdalla M, Schlesinger L, Britigan B . Gallium nitrate is efficacious in murine models of tuberculosis and inhibits key bacterial Fe-dependent enzymes. Antimicrob Agents Chemother. 2013; 57(12):6074-80. PMC: 3837848. DOI: 10.1128/AAC.01543-13. View

Gouzy A, Healy C, Black K, Rhee K, Ehrt S . Growth of at acidic pH depends on lipid assimilation and is accompanied by reduced GAPDH activity. Proc Natl Acad Sci U S A. 2021; 118(32). PMC: 8364206. DOI: 10.1073/pnas.2024571118. View

Rzhepishevska O, Hakobyan S, Ekstrand-Hammarstrom B, Nygren Y, Karlsson T, Bucht A . The gallium(III)-salicylidene acylhydrazide complex shows synergistic anti-biofilm effect and inhibits toxin production by Pseudomonas aeruginosa. J Inorg Biochem. 2014; 138:1-8. DOI: 10.1016/j.jinorgbio.2014.04.009. View

10.

Bonchi C, Imperi F, Minandri F, Visca P, Frangipani E . Repurposing of gallium-based drugs for antibacterial therapy. Biofactors. 2014; 40(3):303-12. DOI: 10.1002/biof.1159. View

11.

Sciscenko I, Arques A, Varga Z, Bouchonnet S, Monfort O, Brigante M . Significant role of iron on the fate and photodegradation of enrofloxacin. Chemosphere. 2021; 270:129791. DOI: 10.1016/j.chemosphere.2021.129791. View

12.

Olakanmi O, Britigan B, Schlesinger L . Gallium disrupts iron metabolism of mycobacteria residing within human macrophages. Infect Immun. 2000; 68(10):5619-27. PMC: 101514. DOI: 10.1128/IAI.68.10.5619-5627.2000. View

13.

Waxman A, Julien P, Brachman M, Tanasescu D, Ramanna L, Birnbaum F . Gallium scintigraphy in bronchogenic carcinoma. The effect of tumor location on sensitivity and specificity. Chest. 1984; 86(2):178-83. DOI: 10.1378/chest.86.2.178. View

14.

Vanino E, Granozzi B, Akkerman O, Munoz-Torrico M, Palmieri F, Seaworth B . Update of drug-resistant tuberculosis treatment guidelines: A turning point. Int J Infect Dis. 2023; 130 Suppl 1:S12-S15. DOI: 10.1016/j.ijid.2023.03.013. View

15.

Rinschen M, Ivanisevic J, Giera M, Siuzdak G . Identification of bioactive metabolites using activity metabolomics. Nat Rev Mol Cell Biol. 2019; 20(6):353-367. PMC: 6613555. DOI: 10.1038/s41580-019-0108-4. View

16.

Riccardi N, Canetti D, Rodari P, Besozzi G, Saderi L, Dettori M . Tuberculosis and pharmacological interactions: A narrative review. Curr Res Pharmacol Drug Discov. 2021; 2:100007. PMC: 8663953. DOI: 10.1016/j.crphar.2020.100007. View

17.

Bernstein L . Mechanisms of therapeutic activity for gallium. Pharmacol Rev. 1998; 50(4):665-82. View

18.

Stokes J, Lopatkin A, Lobritz M, Collins J . Bacterial Metabolism and Antibiotic Efficacy. Cell Metab. 2019; 30(2):251-259. PMC: 6990394. DOI: 10.1016/j.cmet.2019.06.009. View

19.

Bullen J, Rogers H, Spalding P, Ward C . Iron and infection: the heart of the matter. FEMS Immunol Med Microbiol. 2005; 43(3):325-30. DOI: 10.1016/j.femsim.2004.11.010. View

20.

Dietl A, Meir Z, Shadkchan Y, Osherov N, Haas H . Riboflavin and pantothenic acid biosynthesis are crucial for iron homeostasis and virulence in the pathogenic mold Aspergillus fumigatus. Virulence. 2018; 9(1):1036-1049. PMC: 6068542. DOI: 10.1080/21505594.2018.1482181. View