Summary of Study ST002737
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 PR001702. The data can be accessed directly via it's Project DOI: 10.21228/M8Q42J 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 | ST002737 |
| Study Title | 2’-fucosyllactose prevents colitis |
| Study Type | untargeted metabolomics analysis |
| Study Summary | Human milk-derived 2’-fucosyllactose (2’-FL) consumption is associated with health benefits in infancy that extend into adulthood. However, the exact biological functions of 2’-FL and corresponding mechanisms of action remain largely unknown. Here, we investigated the impact of 2’-FL on gut microbial metabolism for the prevention of colitis in adulthood. The gut microbiota from adult mice treated with 2’-FL showed an increase in abundance of several health-associated genera, including Bifidobacterium, and exhibited preventive effects on colitis. Microbial metabolic analysis demonstrated that 26 pathways that are significantly different between non-inflammatory bowel disease individuals and patients with ulcerative colitis (UC) are significantly regulated by 2’-FL in mice, indicating that 2’-FL has the potential to directly regulate dysregulated microbial metabolism in UC. Exploratory metabolomics of Bifidobacterium infantis identified novel secreted metabolites significantly enriched by 2’-FL consumption, including pantothenol. Remarkably, pantothenate significantly protects mucosal barrier and mitigates colitis in adult mice. Thus 2’-FL-modulated gut microbial metabolism may contribute to the prevention of intestinal inflammation in adulthood. |
| Institute | Vanderbilt University |
| Department | Chemistry |
| Laboratory | Center for Innovative Technology |
| Last Name | CODREANU |
| First Name | SIMONA |
| Address | 1234 STEVENSON CENTER LANE |
| SIMONA.CODREANU@VANDERBILT.EDU | |
| Phone | 6158758422 |
| Submit Date | 2023-06-15 |
| Num Groups | 2 |
| Total Subjects | 10 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-12-18 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001702 |
| Project DOI: | doi: 10.21228/M8Q42J |
| Project Title: | 2’-fucosyllactose modulates gut microbial metabolism for the prevention of colitis |
| Project Type: | Untargeted Metabolomics analysis |
| Project Summary: | Human milk-derived 2’-fucosyllactose (2’-FL) consumption is associated with health benefits in infancy that extend into adulthood. However, the exact biological functions of 2’-FL and corresponding mechanisms of action remain largely unknown. Here, we investigated the impact of 2’-FL on gut microbial metabolism for the prevention of colitis in adulthood. The gut microbiota from adult mice treated with 2’-FL showed an increase in abundance of several health-associated genera, including Bifidobacterium, and exhibited preventive effects on colitis. Microbial metabolic analysis demonstrated that 26 pathways that are significantly different between non-inflammatory bowel disease individuals and patients with ulcerative colitis (UC) are significantly regulated by 2’-FL in mice, indicating that 2’-FL has the potential to directly regulate dysregulated microbial metabolism in UC. Exploratory metabolomics of Bifidobacterium infantis identified novel secreted metabolites significantly enriched by 2’-FL consumption, including pantothenol. Remarkably, pantothenate significantly protects mucosal barrier and mitigates colitis in adult mice. Thus 2’-FL-modulated gut microbial metabolism may contribute to the prevention of intestinal inflammation in adulthood. |
| Institute: | Vanderbilt University |
| Department: | Chemistry |
| Laboratory: | Center for Innovative Technology |
| Last Name: | CODREANU |
| First Name: | SIMONA |
| Address: | 1234 STEVENSON CENTER LANE |
| Email: | SIMONA.CODREANU@VANDERBILT.EDU |
| Phone: | 6158758422 |
Subject:
| Subject ID: | SU002844 |
| Subject Type: | Bacteria |
| Subject Species: | Bifidobacterium longum subspecies infantis (B. infantis) |
| Genotype Strain: | ATCC 15702 and ATCC 15697 |
| Cell Biosource Or Supplier: | ATCC |
Factors:
Subject type: Bacteria; Subject species: Bifidobacterium longum subspecies infantis (B. infantis) (Factor headings shown in green)
| mb_sample_id | local_sample_id | treatment |
|---|---|---|
| SA288746 | C_2 | 0 |
| SA288747 | MC_02 | 0 |
| SA288748 | C_1 | 0 |
| SA288749 | MC_03 | 0 |
| SA288750 | C_5 | 0 |
| SA288751 | C_3 | 0 |
| SA288752 | C_4 | 0 |
| SA288753 | MC_01 | 0 |
| SA288754 | MF_03 | 2'FL |
| SA288755 | MF_02 | 2'FL |
| SA288756 | MF_01 | 2'FL |
| SA288757 | F_4 | 2'FL |
| SA288758 | F_1 | 2'FL |
| SA288759 | F_2 | 2'FL |
| SA288760 | F_3 | 2'FL |
| SA288761 | F_5 | 2'FL |
| Showing results 1 to 16 of 16 |
Collection:
| Collection ID: | CO002837 |
| Collection Summary: | B. infantis 15702 were inoculated in RCM at 37C with or without supplementation with 10 mg/mL of 2’-FL (1.0% w/v) until OD600≈1. Bacteria were pelleted and supernatants were filtered (0.2 micron) and stored at -80ºC for untargeted metabolomics. Cultured RCM medium and 10 mg/mL 2’-FL supplemented RCM medium without bacteria inoculation served as controls. Experiments were repeated 2-3 times for collecting five biological replicates from each group. |
| Sample Type: | Bacterial cells |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002853 |
| Treatment Summary: | B. infantis 15702 were inoculated in RCM at 37C with or without supplementation with 10 mg/mL of 2’-FL (1.0% w/v) until OD600≈1. Bacteria were pelleted and supernatants were filtered (0.2 micron) and stored at -80ºC for untargeted metabolomics. Cultured RCM medium and 10 mg/mL 2’-FL supplemented RCM medium without bacteria inoculation served as controls. Experiments were repeated 2-3 times for collecting five biological replicates from each group. |
| Treatment: | 2’-fucosyllactose |
| Treatment Dose: | 10mg/mL |
Sample Preparation:
| Sampleprep ID: | SP002850 |
| Sampleprep Summary: | Bifidobacteria infantis were cultured in Reinforced Clostridial Medium (RCM) with or without 2’-fucosyllactose (2’-FL), a human milk oligosaccharide. Bacteria were pelleted by centrifugation and supernatants were then collected, snap frozen, and stored at -80°C until analyzed via Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS and LC-HRMS/MS)-based metabolomics in the Vanderbilt Center for Innovative Technology using previously described methods. Briefly, equal volumes (200µL) of previously frozen culture medium were prepared. Isotopically labeled standards, biotin-D2 and phenylalanine-D8, were added to individual samples to assess the sample preparation steps. Samples were subjected to protein precipitation by addition of 800µL of ice-cold methanol, then incubated at -80C overnight. Following protein precipitation, samples were centrifuged at 10,000 rpm for 10 min to remove insoluble material. Supernatant(s) were transferred and dried in vacuo, and stored at -80C prior to MS characterization. |
| Processing Storage Conditions: | -80℃ |
| Extraction Method: | Following lysis and standard addition, protein precipitation was performed by adding 800µL of ice-cold methanol (4x by volume). Samples were incubated at -80°C overnight. Following incubation, samples were centrifuged at 10,000 rpm for 10 min to eliminate proteins. The supernatants containing metabolites were dried via speed-vacuum. |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN004439 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Vanquish UHPLC binary system |
| Column | Thermo Hypersil Gold (100 x 2.1mm, 1.9um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | POSITIVE |
| Units | time_m/z |
Chromatography:
| Chromatography ID: | CH003335 |
| Chromatography Summary: | For the RPLC analysis metabolite extracts (5μL injection volume) were separated on a Hypersil Gold, 1.9 µm, 2.1mm x 100 mm column (Thermo Fisher) held at 40°C. Liquid chromatography was performed at a 250μL min-1 using solvent A (0.1% formic acid (FA) in water) and solvent B (0.1% FA in acetonitrile:water 80:20) over a 30 min gradient. Mass spectrometry analyses were performed in positive ion mode with the parameters as previously published except for the following changes. First, tandem mass spectra were acquired using a data dependent scanning mode in which one full MS scan (m/z 70-1050) was followed by 2 or 10 MS/MS scans. MS/MS scans are acquired in profile mode using an isolation width of 1.3 m/z, stepped collision energy (NCE 20, 40), and a dynamic exclusion of 6 s. |
| Instrument Name: | Vanquish UHPLC binary system |
| Column Name: | Thermo Hypersil Gold (100 x 2.1mm, 1.9um) |
| Column Temperature: | 40 |
| Flow Gradient: | 30 min |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | 100% water, 0.1% Formic Acid |
| Solvent B: | 80:20 acetonitrile:water, 0.1% Formic Acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004186 |
| Analysis ID: | AN004439 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The acquired RPLC-HRMS raw data from five (5) biological replicates from each sample type were imported, processed, normalized and reviewed using Progenesis QI v.3.0 (Non-linear Dynamics, Newcastle, UK). All MS and MS/MS sample runs were aligned against a pooled QC reference run. Unique ions (retention time and m/z pairs) were de-adducted and de-isotoped to generate unique “features” (or retention time and m/z pairs). Data were normalized to all features and cleaned by removing spectral features >25% CV in the pooled QC samples. Sample process and instrument variability were also assessed to determine sample acceptance. Briefly, QA metrics for sample process variability and instrument variability are ≤10% CV and ≤5% CV, respectively. In these studies, no samples were identified as outliers. Statistical analyses were performed in Progensis QI using variance stabilized measurements achieved through log normalization to calculate p-values by one-way analysis of variance (ANOVA) test. Significantly changed metabolites were chosen with the criteria p-value <0.05 and |FC| > 2. |
| Ion Mode: | POSITIVE |
