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.
| Study ID | ST003521 |
| Study Title | Metabolic Profiling Unveils Enhanced Antibacterial Synergy of Polymyxin B and Teixobactin against Multi-Drug Resistant Acinetobacter baumannii |
| Study Type | Biomedical research |
| Study 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 |
| maytham.hussein.old@monash.edu | |
| Phone | +61406574736 |
| Submit Date | 2024-10-17 |
| Publications | Metabolic Profiling Unveils Enhanced Antibacterial Synergy of Polymyxin B and Teixobactin against Multi-Drug Resistant Acinetobacter baumannii |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-11-01 |
| Release Version | 1 |
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: | NA |
Factors:
Subject type: Bacteria; Subject species: Acinetobacter baumannii (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | raw file name |
|---|---|---|---|
| SA386726 | COM_1h_1 | Combination_1h | COM_1h_1 |
| SA386727 | COM_1h_2 | Combination_1h | COM_1h_2 |
| SA386728 | COM_1h_3 | Combination_1h | COM_1h_3 |
| SA386729 | COM_1h_4 | Combination_1h | COM_1h_4 |
| SA386730 | COM_3h_1 | Combination_3h | COM_3h_1 |
| SA386731 | COM_3h_2 | Combination_3h | COM_3h_2 |
| SA386732 | COM_3h_3 | Combination_3h | COM_3h_3 |
| SA386733 | COM_3h_4 | Combination_3h | COM_3h_4 |
| SA386734 | COM_6h_1 | Combination_6h | COM_6h_1 |
| SA386735 | COM_6h_2 | Combination_6h | COM_6h_2 |
| SA386736 | COM_6h_3 | Combination_6h | COM_6h_3 |
| SA386737 | COM_6h_4 | Combination_6h | COM_6h_4 |
| SA386738 | Control_1h_1 | Control_1h | Control_1h_1 |
| SA386739 | Control_1h_2 | Control_1h | Control_1h_2 |
| SA386740 | Control_1h_3 | Control_1h | Control_1h_3 |
| SA386741 | Control_1h_4 | Control_1h | Control_1h_4 |
| SA386742 | Control_3h_1 | Control_3h | Control_3h_1 |
| SA386743 | Control_3h_2 | Control_3h | Control_3h_2 |
| SA386744 | Control_3h_3 | Control_3h | Control_3h_3 |
| SA386745 | Control_3h_4 | Control_3h | Control_3h_4 |
| SA386746 | Control_6h_1 | Control_6h | Control_6h_1 |
| SA386747 | Control_6h_2 | Control_6h | Control_6h_2 |
| SA386748 | Control_6h_3 | Control_6h | Control_6h_3 |
| SA386749 | Control_6h_4 | Control_6h | Control_6h_4 |
| SA386750 | Blank_1 | Extraction blank | Blank_1 |
| SA386751 | Blank_2 | Extraction blank | Blank_2 |
| SA386752 | Blank_3 | Extraction blank | Blank_3 |
| SA386753 | Blank_4 | Extraction blank | Blank_4 |
| SA386754 | Blank_5 | Extraction blank | Blank_5 |
| SA386755 | TX_1h_1 | Leu10-teixobactin_1h | TX_1h_1 |
| SA386756 | TX_1h_2 | Leu10-teixobactin_1h | TX_1h_2 |
| SA386757 | TX_1h_3 | Leu10-teixobactin_1h | TX_1h_3 |
| SA386758 | TX_1h_4 | Leu10-teixobactin_1h | TX_1h_4 |
| SA386759 | TX_3h_1 | Leu10-teixobactin_3h | TX_3h_1 |
| SA386760 | TX_3h_2 | Leu10-teixobactin_3h | TX_3h_2 |
| SA386761 | TX_3h_3 | Leu10-teixobactin_3h | TX_3h_3 |
| SA386762 | TX_3h_4 | Leu10-teixobactin_3h | TX_3h_4 |
| SA386763 | TX_6h_1 | Leu10-teixobactin_6h | TX_6h_1 |
| SA386764 | TX_6h_2 | Leu10-teixobactin_6h | TX_6h_2 |
| SA386765 | TX_6h_3 | Leu10-teixobactin_6h | TX_6h_3 |
| SA386766 | TX_6h_4 | Leu10-teixobactin_6h | TX_6h_4 |
| SA386767 | PMB_1h_1 | Polymyxin B_1h | PMB_1h_1 |
| SA386768 | PMB_1h_2 | Polymyxin B_1h | PMB_1h_2 |
| SA386769 | PMB_1h_3 | Polymyxin B_1h | PMB_1h_3 |
| SA386770 | PMB_1h_4 | Polymyxin B_1h | PMB_1h_4 |
| SA386771 | PMB_3h_1 | Polymyxin B_3h | PMB_3h_1 |
| SA386772 | PMB_3h_2 | Polymyxin B_3h | PMB_3h_2 |
| SA386773 | PMB_3h_3 | Polymyxin B_3h | PMB_3h_3 |
| SA386774 | PMB_3h_4 | Polymyxin B_3h | PMB_3h_4 |
| SA386775 | PMB_6h_1 | Polymyxin B_6h | PMB_6h_1 |
| SA386776 | PMB_6h_2 | Polymyxin B_6h | PMB_6h_2 |
| SA386777 | PMB_6h_3 | Polymyxin B_6h | PMB_6h_3 |
| SA386778 | PMB_6h_4 | Polymyxin B_6h | PMB_6h_4 |
| SA386779 | QC_1 | Pooled QC | QC_1 |
| SA386780 | QC_2 | Pooled QC | QC_2 |
| SA386781 | QC_3 | Pooled QC | QC_3 |
| SA386782 | QC_4 | Pooled QC | QC_4 |
| SA386783 | QC_5 | Pooled 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 |
