Summary of Study ST004228
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 PR002668. The data can be accessed directly via it's Project DOI: 10.21228/M8SG2M 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 | ST004228 |
| Study Title | Aromatic Microbial Metabolite Hippuric Acid Potentiates Pro-Inflammatory Responses in Macrophages through TLR-MyD88 Signaling and Lipid Remodeling - Non-targeted metabolomics of C57BL/6 mice infected with Escherichia coli |
| Study Summary | Authors have investigated changes in microbial metabolites during bacterial infection. In this study, C57BL/6 mice were infected intraperitoneally with Escherichia coli (2 × 10⁷ CFU), and non-targeted metabolomics using liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed on sera collected 48 hours post-infection. The metabolomics screen revealed significant shifts in metabolic profiles, including altered levels of 2/3-hydroxybutyrate (a marker of fatty acid metabolism), hypoxanthine (a marker of purine metabolism), and N-acetylglutamine (involved in nitrogen metabolism), indicating broad metabolic reprogramming during infection. Among the most significantly altered metabolites, hippuric acid—an aromatic compound produced by microbial metabolism of dietary polyphenols and amino acids was markedly depleted, showing a 24-fold reduction in infected mice compared to uninfected controls (p < 0.001). These findings establish an association between bacterial infection and the depletion of hippuric acid, indicating its role in modulating immune responses during infection. |
| Institute | The Wistar Institute |
| Last Name | Shinde |
| First Name | Rahul |
| Address | 3601 Spruce Street, Philadelphia, PA, 19104, USA |
| rshinde@wistar.org | |
| Phone | 215-898-3717 |
| Submit Date | 2025-09-18 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-10-31 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002668 |
| Project DOI: | doi: 10.21228/M8SG2M |
| Project Title: | Aromatic Microbial Metabolite Hippuric Acid Potentiates Pro-Inflammatory Responses in Macrophages through TLR-MyD88 Signaling and Lipid Remodeling |
| Project Summary: | The gut microbiome generates a diverse array of metabolites that actively shape host immunity, yet the pro-inflammatory potential of microbial metabolites remains incompletely understood. Using a non-targeted, liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based metabolomics, we identified hippuric acid, an aromatic gut microbe-derived metabolite, as a potent enhancer of pro-inflammatory responses in Escherichia coli infection model. Intraperitoneal administration of hippuric acid significantly heightened pro-inflammatory responses, promoted innate immune cell activation, and reduced survival in infected mice. Similar pro-inflammatory effects were observed in an LPS-induced inflammation model. In vitro, hippuric acid selectively potentiated M1-like macrophage polarization (LPS + IFNγ) but had no effect on M2-like polarization (IL-4). Hippuric acid further augmented responses to multiple myeloid differentiation primary response 88 (MyD88)-dependent toll-like receptor (TLR) ligands, but not to TRIF-dependent TLR3, or to cytosolic innate immune stimuli such as STING and NOD2 agonists, implicating TLR-MyD88 signaling as a likely mechanism of action. Genetic deletion of MyD88 abrogated the pro-inflammatory effects of hippuric acid both in vitro and in vivo, confirming its dependence on the MyD88 pathway. Transcriptomic and lipidomic analyses revealed that hippuric acid upregulated cholesterol biosynthesis and induced lipid accumulation. Pharmacological reduction of cellular cholesterol using fluvastatin or 25-hydroxycholesterol attenuated its pro-inflammatory effects. Notably, hippuric acid also enhanced pro-inflammatory responses in human macrophages, and its elevated levels correlated with increased sepsis mortality, underscoring its clinical relevance. Together, these findings identify hippuric acid as a previously unrecognized microbial-derived pro-inflammatory modulator that links gut microbial metabolism, lipid remodeling, and innate immune signaling, and offer new insights into its role in infection and inflammation. |
| Institute: | The Wistar Institute |
| Last Name: | Shinde |
| First Name: | Rahul |
| Address: | 3601 Spruce Street, Philadelphia, PA, 19104, USA |
| Email: | rshinde@wistar.org |
| Phone: | 215-898-3717 |
Subject:
| Subject ID: | SU004380 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment | Sample source |
|---|---|---|---|
| SA486086 | C1 | Control | Serum |
| SA486087 | C2 | Control | Serum |
| SA486088 | C3 | Control | Serum |
| SA486089 | C4 | Control | Serum |
| SA486090 | C5 | Control | Serum |
| SA486091 | E1 | E. coli | Serum |
| SA486092 | E2 | E. coli | Serum |
| SA486093 | E3 | E. coli | Serum |
| SA486094 | E4 | E. coli | Serum |
| SA486095 | E5 | E. coli | Serum |
| SA486096 | QC-6 | n/a | Serum |
| SA486097 | QC-7 | n/a | Serum |
| SA486098 | QC-8 | n/a | Serum |
| SA486099 | QC-9 | n/a | Serum |
| Showing results 1 to 14 of 14 |
Collection:
| Collection ID: | CO004373 |
| Collection Summary: | Serum samples were collected 48 hours post-infection |
| Sample Type: | Blood (serum) |
Treatment:
| Treatment ID: | TR004389 |
| Treatment Summary: | Escherichia coli peritoneal infection was performed as previously described. Briefly, E. coli (ATCC 25922) was cultured in LB medium with 100 µg/mL ampicillin overnight at 37 °C with gentle shaking to an optical density at wavelength 600 nm of 0.5−0.6 to achieve a logarithmic growth phase. C57BL/6J or MyD88 KO mice of 8–12 weeks of age received an intraperitoneal (i.p.) injection of 100 μl PBS with 2 × 10^7 E. coli. |
Sample Preparation:
| Sampleprep ID: | SP004386 |
| Sampleprep Summary: | Polar metabolites were extracted from serum samples with 10-times the volume of ice-cold 80% methanol spiked with 1 μM HA-13C6 as an internal standard for HA in isotope dilution MS. |
Chromatography:
| Chromatography ID: | CH005343 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | SeQuant ZIC- pHILIC (150 x 2.1mm,5um) |
| Column Temperature: | 45 |
| Flow Gradient: | 85% B for 2 min, 85% B to 20% B over 15 min, 20% B to 85% B over 0.1 min, and 85% B for 8.9 min |
| Flow Rate: | 0.2 ml/min |
| Solvent A: | 100% water; 20 mM ammonium carbonate; 5 µM medronic acid; 0.1% ammonium hydroxide; pH 9.2 |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
Analysis:
| Analysis ID: | AN007037 |
| Analysis Type: | MS |
| Chromatography ID: | CH005343 |
| Num Factors: | 3 |
| Num Metabolites: | 95 |
| Units: | Peak Area |
| Analysis ID: | AN007038 |
| Analysis Type: | MS |
| Chromatography ID: | CH005343 |
| Num Factors: | 3 |
| Num Metabolites: | 99 |
| Units: | Peak Area |
