Summary of Study ST002747
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 PR001710. The data can be accessed directly via it's Project DOI: 10.21228/M8P43M 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 | ST002747 |
Study Title | Evolutionary genomics identifies host-directed therapeutics to treat intracellular bacterial infections |
Study Summary | Obligate intracellular bacteria from the Rickettsiaceae family have shed essential biosynthetic pathways during their evolution towards host dependency. By systematically comparing this cytosolic family of bacteria to the related vacuolar Anaplasmataceae family using a novel computational pipeline called PoMeLo, we identified 20 metabolic pathways that may have been lost since the divergence of Anaplasmataceae and Rickettsiaceae, corresponding to the latter’s change to a cytosolic niche. We hypothesized that drug inhibition of these host metabolic pathways would reduce the levels of metabolic products available to the bacteria, thereby inhibiting bacterial growth. We tested 22 commercially available inhibitors for 14 of the identified pathways and found that 59% of the inhibitors reduced bacterial growth at concentrations that did not contribute to host cell cytotoxicity. Of these, 5 inhibitors with an IC50 under 5 µM were tested to determine whether their mode of inhibition was bactericidal or bacteriostatic. Both mycophenolate mofetil, an inhibitor of inosine-5'-monophosphate dehydrogenase in the purine biosynthesis pathway, and roseoflavin, an analog of riboflavin, displayed bactericidal activity. We then took an unbiased metabolomics approach to Rickettsia-infected cells to determine whether there was any overlap between our predicted host pathways and depletion of metabolite levels in infected cells, as measured by mass spectrometry. Our results show that 13 pathways were identified as metabolic gaps in both our computational predictions and our metabolomics analysis. These in vitro validation studies support the feasibility of a novel evolutionary genomics-guided approach for antibiotic drug development against obligate pathogens. |
Institute | CZ Biohub |
Last Name | DeFelice |
First Name | Brian |
Address | 1291 Welch Rd., Rm. G0821 (SIM1), Stanford CA, California, 94305, USA |
bcdefelice@ucdavis.edu | |
Phone | 5303564485 |
Submit Date | 2023-06-23 |
Raw Data Available | Yes |
Raw Data File Type(s) | mzML, raw(Thermo) |
Analysis Type Detail | LC-MS |
Release Date | 2023-07-07 |
Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
Analysis ID | AN004454 | AN004455 | AN004456 | AN004457 |
---|---|---|---|---|
Analysis type | MS | MS | MS | MS |
Chromatography type | HILIC | HILIC | Reversed phase | Reversed phase |
Chromatography system | Thermo Vanquish | Thermo Vanquish | Thermo Vanquish | Thermo Vanquish |
Column | Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um) | Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um) | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) |
MS Type | ESI | ESI | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
Ion Mode | POSITIVE | NEGATIVE | POSITIVE | NEGATIVE |
Units | counts, height | counts, height | counts, height | counts, height |
Chromatography:
Chromatography ID: | CH003345 |
Chromatography Summary: | HILIC |
Instrument Name: | Thermo Vanquish |
Column Name: | Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um) |
Column Temperature: | 45C |
Flow Gradient: | Gradient elution was performed from 100% (B) at 0–2 min to 70% (B) at 7.7 min, 40% (B) at 9.5 min, 30% (B) at 10.25 min, 100% (B) at 12.75 min, isocratic until 16.75 min with a column flow of |
Flow Rate: | 0.4 mL/min. |
Solvent A: | 100% water; 10mM ammonium formate; 0.125% formic acid |
Solvent B: | 95% acetonitrile; 10mM ammonium formate; 0.125% formic acid |
Chromatography Type: | HILIC |
Chromatography ID: | CH003346 |
Chromatography Summary: | Lipids (ammonium acetate replaces ammonium formate in neg mode analysis |
Instrument Name: | Thermo Vanquish |
Column Name: | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) |
Column Temperature: | 65C |
Flow Gradient: | Gradient elution was performed from 15% (B) at 0 min to 30% (B) at 2 min, 48% (B) at 2.5 min, 82% (B) at 11 min, 99% (B) at 11.5 min, isocratic until 12 and resetting to initial conditions at 12.1 min through the end of the run at 15 min. |
Flow Rate: | is 0.6 mL/min. |
Solvent A: | 60% acetonitrile; 10 mM ammonium formate; 0.1% formic acid |
Solvent B: | 10% acetonitrile/90% isopropanol; 10 mM ammonium formate; 0.1% formic acid |
Chromatography Type: | Reversed phase |