Summary of Study ST003521

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002166. The data can be accessed directly via it's Project DOI: 10.21228/M8N829 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST003521
Study TitleMetabolic Profiling Unveils Enhanced Antibacterial Synergy of Polymyxin B and Teixobactin against Multi-Drug Resistant Acinetobacter baumannii
Study TypeBiomedical research
Study SummaryThis untargeted metabolomics study investigated the synergistic antibacterial activity of polymyxin B and Leu10-teixobactin, a depsipeptide inhibitor of cell wall biosynthesis. Checkerboard microdilution assays revealed a significant synergy against polymyxin-susceptible and -resistant A. baumannii, excluding lipopolysaccharide-deficient variants. Time-kill assays confirmed bactericidal synergy, reducing bacterial burden by approximately 4-6-log10CFU/mL. The combination (2xMIC polymyxin B and 0.5xMIC Leu10-teixobactin) prevented bacterial regrowth after 24 h, indicating sustained efficacy against the emergence of resistant mutants. The analysis of A. baumannii ATCCTM 19606 metabolome demonstrated that the polymyxin B–Leu10-teixobactin combination produced more pronounced perturbation compared to the individual antibiotics across all time points (1, 3 and 6 h). Pathway analysis revealed that lipid metabolism, cell envelope biogenesis, and cellular respiration were predominantly impacted by the combination, and to a lesser extent by polymyxin B monotherapy. Leu10-teixobactin treatment alone had only a minor impact on the metabolome, primarily at the 6 h time point. Peptidoglycan assays confirmed the combination’s concerted deleterious effects on bacterial cell envelope integrity. Electron microscopy further substantiated these findings, revealing pronounced cell envelope damage, membrane blebbing, and vacuole formation. These findings highlight the potential of the polymyxin B–Leu10-teixobactin combination as an effective treatment in preventing resistance in A. baumannii.
Institute
Monash University
DepartmentPharmacology
LaboratoryVelkov
Last NameHUSSEIN
First NameMAYTHAM
Address9 Ancora Imparo Way, Building 13E, Monash University
Emailmaytham.hussein.old@monash.edu
Phone+61406574736
Submit Date2024-10-17
PublicationsMetabolic Profiling Unveils Enhanced Antibacterial Synergy of Polymyxin B and Teixobactin against Multi-Drug Resistant Acinetobacter baumannii
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2024-11-01
Release Version1
MAYTHAM HUSSEIN MAYTHAM HUSSEIN
https://dx.doi.org/10.21228/M8N829
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:


Project:

Project ID:PR002166
Project DOI:doi: 10.21228/M8N829
Project Title:Metabolic Profiling Unveils Enhanced Antibacterial Synergy of Polymyxin B and Teixobactin against Multi-Drug Resistant Acinetobacter baumannii
Project Type:Untargeted metabolomics
Project Summary:This untargeted metabolomics study investigated the synergistic antibacterial activity of polymyxin B and Leu10-teixobactin, a depsipeptide inhibitor of cell wall biosynthesis. Checkerboard microdilution assays revealed a significant synergy against polymyxin-susceptible and -resistant A. baumannii, excluding lipopolysaccharide-deficient variants. Time-kill assays confirmed bactericidal synergy, reducing bacterial burden by approximately 4-6-log10CFU/mL. The combination (2xMIC polymyxin B and 0.5xMIC Leu10-teixobactin) prevented bacterial regrowth after 24 h, indicating sustained efficacy against the emergence of resistant mutants. The analysis of A. baumannii ATCCTM 19606 metabolome demonstrated that the polymyxin B–Leu10-teixobactin combination produced more pronounced perturbation compared to the individual antibiotics across all time points (1, 3 and 6 h). Pathway analysis revealed that lipid metabolism, cell envelope biogenesis, and cellular respiration were predominantly impacted by the combination, and to a lesser extent by polymyxin B monotherapy. Leu10-teixobactin treatment alone had only a minor impact on the metabolome, primarily at the 6 h time point. Peptidoglycan assays confirmed the combination’s concerted deleterious effects on bacterial cell envelope integrity. Electron microscopy further substantiated these findings, revealing pronounced cell envelope damage, membrane blebbing, and vacuole formation. These findings highlight the potential of the polymyxin B–Leu10-teixobactin combination as an effective treatment in preventing resistance in A. baumannii.
Institute:Monash University
Department:Pharmacology
Laboratory:Velkov
Last Name:Hussein
First Name:Maytham
Address:9 Ancora Imparo Way, Building 13E, Monash University
Email:maytham.hussein.old@monash.edu
Phone:+61406574736
Publications:Metabolic Profiling Unveils Enhanced Antibacterial Synergy of Polymyxin B and Teixobactin against Multi-Drug Resistant Acinetobacter baumannii
Contributors:Maytham Hussein,1* Zhisen Kang,1 Stephanie L. Neville,2 Rafah Allobawi,1 Varsha Thrombare,1 Augustine Jing Jie Koh,1,3 Jonathan Wilksch,2 Simon Crawford,4 Mudher Khudhur Mohammed,5 Christopher A. McDevitt,2 Mark Baker,6 Gauri G. Rao,7* Jian Li,4* Tony Velkov1*

Subject:

Subject ID:SU003650
Subject Type:Bacteria
Subject Species:Acinetobacter baumannii
Taxonomy ID:509173
Genotype Strain:Acinetobacter baumannii ATCC 19606
Age Or Age Range:NA
Weight Or Weight Range:NA
Height Or Height Range:NA
Gender:Not applicable
Cell Biosource Or Supplier:NA
Cell Strain Details:NA
Subject Comments:NA
Cell Primary Immortalized:NA
Cell Passage Number:NA
Cell Counts:NA
Species Group:Bacteria

Factors:

Subject type: Bacteria; Subject species: Acinetobacter baumannii (Factor headings shown in green)

mb_sample_id local_sample_id Sample source raw file name
SA386726COM_1h_1Combination_1h COM_1h_1
SA386727COM_1h_2Combination_1h COM_1h_2
SA386728COM_1h_3Combination_1h COM_1h_3
SA386729COM_1h_4Combination_1h COM_1h_4
SA386730COM_3h_1Combination_3h COM_3h_1
SA386731COM_3h_2Combination_3h COM_3h_2
SA386732COM_3h_3Combination_3h COM_3h_3
SA386733COM_3h_4Combination_3h COM_3h_4
SA386734COM_6h_1Combination_6h COM_6h_1
SA386735COM_6h_2Combination_6h COM_6h_2
SA386736COM_6h_3Combination_6h COM_6h_3
SA386737COM_6h_4Combination_6h COM_6h_4
SA386738Control_1h_1Control_1h Control_1h_1
SA386739Control_1h_2Control_1h Control_1h_2
SA386740Control_1h_3Control_1h Control_1h_3
SA386741Control_1h_4Control_1h Control_1h_4
SA386742Control_3h_1Control_3h Control_3h_1
SA386743Control_3h_2Control_3h Control_3h_2
SA386744Control_3h_3Control_3h Control_3h_3
SA386745Control_3h_4Control_3h Control_3h_4
SA386746Control_6h_1Control_6h Control_6h_1
SA386747Control_6h_2Control_6h Control_6h_2
SA386748Control_6h_3Control_6h Control_6h_3
SA386749Control_6h_4Control_6h Control_6h_4
SA386750Blank_1Extraction blank Blank_1
SA386751Blank_2Extraction blank Blank_2
SA386752Blank_3Extraction blank Blank_3
SA386753Blank_4Extraction blank Blank_4
SA386754Blank_5Extraction blank Blank_5
SA386755TX_1h_1Leu10-teixobactin_1h TX_1h_1
SA386756TX_1h_2Leu10-teixobactin_1h TX_1h_2
SA386757TX_1h_3Leu10-teixobactin_1h TX_1h_3
SA386758TX_1h_4Leu10-teixobactin_1h TX_1h_4
SA386759TX_3h_1Leu10-teixobactin_3h TX_3h_1
SA386760TX_3h_2Leu10-teixobactin_3h TX_3h_2
SA386761TX_3h_3Leu10-teixobactin_3h TX_3h_3
SA386762TX_3h_4Leu10-teixobactin_3h TX_3h_4
SA386763TX_6h_1Leu10-teixobactin_6h TX_6h_1
SA386764TX_6h_2Leu10-teixobactin_6h TX_6h_2
SA386765TX_6h_3Leu10-teixobactin_6h TX_6h_3
SA386766TX_6h_4Leu10-teixobactin_6h TX_6h_4
SA386767PMB_1h_1Polymyxin B_1h PMB_1h_1
SA386768PMB_1h_2Polymyxin B_1h PMB_1h_2
SA386769PMB_1h_3Polymyxin B_1h PMB_1h_3
SA386770PMB_1h_4Polymyxin B_1h PMB_1h_4
SA386771PMB_3h_1Polymyxin B_3h PMB_3h_1
SA386772PMB_3h_2Polymyxin B_3h PMB_3h_2
SA386773PMB_3h_3Polymyxin B_3h PMB_3h_3
SA386774PMB_3h_4Polymyxin B_3h PMB_3h_4
SA386775PMB_6h_1Polymyxin B_6h PMB_6h_1
SA386776PMB_6h_2Polymyxin B_6h PMB_6h_2
SA386777PMB_6h_3Polymyxin B_6h PMB_6h_3
SA386778PMB_6h_4Polymyxin B_6h PMB_6h_4
SA386779QC_1Pooled QC QC_1
SA386780QC_2Pooled QC QC_2
SA386781QC_3Pooled QC QC_3
SA386782QC_4Pooled QC QC_4
SA386783QC_5Pooled QC QC_5
Showing results 1 to 58 of 58

Collection:

Collection ID:CO003643
Collection Summary:An untargeted metabolomics study was carried out to explore the synergistic antibacterial activity of polymyxin B and Leu10-teixobactin against A. baumannii ATCCTM 19606 using a concentration of 1 mg/L for polymyxin B, 4 mg/L for Leu10-teixobactin. Samples were taken and analyzed at the 1-, 3-, and 6-h time points in 4 biological replicates. An overnight culture was prepared by inoculating a single colony into 100 mL CAMHB in 250 mL conical flasks (Pyrex) and incubating the suspension in a shaker at 37°C and 180 rpm for ~16 h. After incubation overnight, log-phase cells were prepared in fresh MHB and then incubated for 2 h at 37°C at 180 rpm to log phase with a starting bacterial inoculum of 108 CFU/mL.
Sample Type:Bacterial cells

Treatment:

Treatment ID:TR003659
Treatment Summary:Stock solutions of polymyxin B, Leu10-teixobactin or their combination were added accordingly to three treatment flasks, with final concentrations of 1 mg/L for polymyxin B, 4 mg/L for Leu10-teixobactin, and the respective combination concentrations; the fourth flask served as the untreated control. The flasks were then incubated at 37°C with shaking at 180 rpm. At each time point (1, 3, and 6 h), 15-mL samples were transferred to 50-mL Falcon tubes for quenching, and the optical density reading at 600 nm (OD600) was then measured and normalized to the pre-treatment level of approximately ~0.5 with fresh CAMHB. Samples were then centrifuged at 3,220 g and 4°C for 10 min, and the supernatants were removed. The pellets were stored at -80°C until metabolite extraction. The experiment was performed in 4 biological replicates to reduce the bias from inherent random variation.
Treatment Dose:1mg/L polymyxin B;4mg/L Leu10-teixobactin
Cell Media:Cation-Adjusted Mueller–Hinton Broth (CAMHB)

Sample Preparation:

Sampleprep ID:SP003657
Sampleprep Summary:The bacterial pellets were washed twice in 1 mL of 0.9% saline and then centrifuged at 3,220 g and 4°C for 5 min to remove residual extracellular metabolites and medium components. The washed pellets were resuspended in a cold extraction solvent (chloroform-methanol-water at 1:3:1, vol/vol) containing 1 µM each of the internal standards 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), piperazine-N, N-bis (2-ethanesulfonic acid) (PIPES), and Tris. The samples were then frozen in liquid nitrogen, thawed on ice, and vortexed to release the intracellular metabolites (3 times). Next, the samples were transferred to 1.5-mL Eppendorf tubes and centrifuged at 14,000 g at 4°C for 10 min to remove any particulate matter. Finally, 200 µL of the supernatant was transferred into injection vials for liquid chromatography-mass spectrometry (LC-MS) analysis. An equal volume of each sample was combined and used as a quality control (QC) sample

Combined analysis:

Analysis ID AN005782 AN005783
Analysis type MS MS
Chromatography type HILIC HILIC
Chromatography system Thermo Dionex Ultimate 3000 Thermo Dionex Ultimate 3000
Column SeQuant ZIC-HILIC (150 x 4.6mm,3.5um) SeQuant ZIC-HILIC (150 x 4.6mm,3.5um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive Orbitrap Thermo Q Exactive Orbitrap
Ion Mode POSITIVE NEGATIVE
Units peak height peak height

Chromatography:

Chromatography ID:CH004389
Methods Filename:Metabolomics_pHILIC_Parkville_v1.pdf
Instrument Name:Thermo Dionex Ultimate 3000
Column Name:SeQuant ZIC-HILIC (150 x 4.6mm,3.5um)
Column Pressure:60 bar at starting conditions. 180 bar at %A
Column Temperature:4 C
Flow Gradient:0 min - 80%B, 15 min - 50%B, 18 min - 5%B, 21 min - 5%B, 24 min - 80%B, 32 min - 80%B
Flow Rate:0.3 ml/min
Injection Temperature:4 C
Internal Standard:CAPS, CHAPS, PIPES
Solvent A:100% water; 20 mM ammonium carbonate
Solvent B:100% acetonitrile
Analytical Time:32 min
Capillary Voltage:3.5 kV
Oven Temperature:25 C
Washing Buffer:syringe wash 50% IPA
Weak Wash Solvent Name:50% IPA
Strong Wash Solvent Name:50% IPA
Sample Loop Size:25 uL
Sample Syringe Size:25 uL
Chromatography Type:HILIC

MS:

MS ID:MS005502
Analysis ID:AN005782
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Samples were injected into a nano-high-performance liquid chromatography (nano-HPLC) system (Dionex UltiMate 3000 RSLCnano System; Thermo Fisher Scientific), equipped with a ZIC-pHILIC column (SeQuant, 5 μm, polymeric, 150 × 4.6 mm; Merck), and coupled to a Q-Exactive Orbitrap mass spectrometer (Thermo Fisher Scientific, Australia). The mass spectrometer operated in positive/negative ion–switching electron-spray ionisation (ESI) mode, with a full scan range set at 85-1275 m/z and a resolution of 35,000.
Ion Mode:POSITIVE
  
MS ID:MS005503
Analysis ID:AN005783
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Samples were injected into a nano-high-performance liquid chromatography (nano-HPLC) system (Dionex UltiMate 3000 RSLCnano System; Thermo Fisher Scientific), equipped with a ZIC-pHILIC column (SeQuant, 5 μm, polymeric, 150 × 4.6 mm; Merck), and coupled to a Q-Exactive Orbitrap mass spectrometer (Thermo Fisher Scientific, Australia). The mass spectrometer operated in positive/negative ion–switching electron-spray ionisation (ESI) mode, with a full scan range set at 85-1275 m/z and a resolution of 35,000.
Ion Mode:NEGATIVE
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