Summary of Study ST004132
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 PR002599. The data can be accessed directly via it's Project DOI: 10.21228/M8PZ5N 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 | ST004132 |
| Study Title | Metabolite profiling of portal plasma, cardiac puncture plasma, and cecal contents of 129 T, 129 J, C57Bl6 mice. |
| Study Summary | This study compared portal and cardiac blood metabolomes in B6J, 129J, and 129T mice on a high-fat diet to integrate host genetic effects, gut microbiome profiles, and portal metabolite enrichment. While 27 metabolites were consistently enriched in portal blood and 24 in cardiac blood across all strains, most metabolites showed strain-specific enrichment patterns. Examples include stearate being portal-enriched only in B6J mice, acetyl-galactosamine in 129 strains, and C4 carnitine cardiac-enriched in B6J but not in 129 strains. These differences were not strongly correlated with cecal metabolite profiles, highlighting the role of distinct gut regions and host–microbiome interactions in shaping portal metabolite composition. Correlation analyses revealed that metabolites enriched in 129 strains were generally associated with metabolically beneficial (Class I+) bacteria and favorable metabolic traits, while those enriched in B6J mice correlated with metabolically harmful (Class II+) bacteria and adverse metabolic outcomes. Furthermore, specific metabolites displayed strain-dependent shifts in portal versus cardiac enrichment, suggesting differences in hepatic extraction or addition. Overall, higher portal enrichment in 129 strains was linked to improved metabolic profiles, whereas higher enrichment in B6J mice was associated with metabolic dysfunction. |
| Institute | Broad Institute of MIT and Harvard |
| Last Name | Kahn |
| First Name | Ronald |
| Address | One Joslin Place, Boston, MA 02215 |
| c.ronald.kahn@joslin.harvard.edu | |
| Phone | (617) 309-2635 |
| Submit Date | 2025-08-15 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-09-02 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002599 |
| Project DOI: | doi: 10.21228/M8PZ5N |
| Project Title: | Portal Vein Enriched Metabolites as Intermediate Regulators of the Gut Microbiome in Insulin Resistance |
| Project Summary: | Diet and obesity significantly contribute to insulin resistance and type 2 diabetes, in part via the gut microbiome. To explore the role of gut-derived metabolites on host metabolism, we assessed portal/peripheral blood metabolites in mice with different risks of obesity/diabetes challenged with high-fat diet (HFD) +/- antibiotics. In diabetes/obesity-prone C57Bl/6J mice, 111 were portally-enriched and 74 were peripherally-enriched, many of which differed in metabolic-syndrome-resistant 129S1/129S6 mice. Vancomycin treatment of HFD-fed C57Bl/6J mice modified the microbiome and portal/peripheral ratio of many metabolites, including upregulating multiple TCA cycle-related metabolites, like mesaconate, in portal blood. Treatment of hepatocytes in vitro with mesaconate and its isomers (itaconate, citraconate) improved insulin signaling and transcriptionally regulated genes involved in gluconeogenesis, fatty acid oxidation and lipogenesis both in vitro and in vivo. Thus, portal vs. peripheral metabolites play important roles in mediating effects of the microbiome on hepatic metabolism and the pathogenesis of HFD-related insulin resistance. This data deposition contains two studies: one assessing the role of diet and the microbiome on portal metabolites, and another estimating the role of genetic background on portal-enriched metabolites. |
| Institute: | Broad Institute of MIT and Harvard |
| Department: | Metabolomics Platform |
| Last Name: | Kahn |
| First Name: | Ronald |
| Address: | One Joslin Place, Boston, MA 02215 |
| Email: | c.ronald.kahn@joslin.harvard.edu |
| Phone: | (617) 309-2635 |
| Funding Source: | R01DK121967, R01DK031036, R01DK128429, P30DK036836 (to Joslin Diabetes Center), the Mary K. Iacocca Professorship (to C.R.K), and the FAPESP Process No.: 2022/05957-0 |
| Project Comments: | Two part study (Part 1) |
| Contributors: | Vitor Rosetto Muñoz, Francois Moreau, Marion Soto, Loc Duyen Pham, Jimmy Zhong, Sam Zimmerman, Bruna B. Brandao, Khyati Girdhar, Julian Avila, Hui Pan, Jonathan Dreyfuss, Emrah Altindis, Aleksandar Kostic, Clary B. Clish, C. Ronald Kahn |
Subject:
| Subject ID: | SU004281 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | B6J, 129S1, and 129S6 (129T) |
| Age Or Age Range: | 8 week, 20 weeks at end of treatment |
| Gender: | Male |
| Animal Animal Supplier: | B6J and 129J from Jackson Laboratory (Bar Harbor, ME) and 129T from Taconic Farms (Germantown, NY) |
| Animal Housing: | Housed 4 animals per cage |
| Animal Feed: | High fat diet containing 60% fat by calories (HFD, D12492, Research Diets) |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Strain | Injection order |
|---|---|---|---|---|
| SA476675 | CP14 | Cardiac Puncture | 129 J | 11 |
| SA476676 | CP17 | Cardiac Puncture | 129 J | 12 |
| SA476677 | CP18 | Cardiac Puncture | 129 J | 13 |
| SA476678 | CP16 | Cardiac Puncture | 129 J | 17 |
| SA476679 | CP13 | Cardiac Puncture | 129 J | 20 |
| SA476680 | CP15 | Cardiac Puncture | 129 J | 7 |
| SA476681 | CP7 | Cardiac Puncture | 129 T | 10 |
| SA476682 | CP12 | Cardiac Puncture | 129 T | 14 |
| SA476683 | CP9 | Cardiac Puncture | 129 T | 15 |
| SA476684 | CP8 | Cardiac Puncture | 129 T | 6 |
| SA476685 | CP10 | Cardiac Puncture | 129 T | 8 |
| SA476686 | CP11 | Cardiac Puncture | 129 T | 9 |
| SA476687 | CP6 | Cardiac Puncture | C57Bl6 | 16 |
| SA476688 | CP3 | Cardiac Puncture | C57Bl6 | 18 |
| SA476689 | CP1 | Cardiac Puncture | C57Bl6 | 19 |
| SA476690 | CP5 | Cardiac Puncture | C57Bl6 | 3 |
| SA476691 | CP4 | Cardiac Puncture | C57Bl6 | 4 |
| SA476692 | CP2 | Cardiac Puncture | C57Bl6 | 5 |
| SA476693 | CC15 | Cecal Contents | 129 J | 51 |
| SA476694 | CC14 | Cecal Contents | 129 J | 55 |
| SA476695 | CC17 | Cecal Contents | 129 J | 56 |
| SA476696 | CC18 | Cecal Contents | 129 J | 57 |
| SA476697 | CC16 | Cecal Contents | 129 J | 61 |
| SA476698 | CC13 | Cecal Contents | 129 J | 64 |
| SA476699 | CC8 | Cecal Contents | 129 T | 50 |
| SA476700 | CC10 | Cecal Contents | 129 T | 52 |
| SA476701 | CC11 | Cecal Contents | 129 T | 53 |
| SA476702 | CC7 | Cecal Contents | 129 T | 54 |
| SA476703 | CC12 | Cecal Contents | 129 T | 58 |
| SA476704 | CC9 | Cecal Contents | 129 T | 59 |
| SA476705 | CC5 | Cecal Contents | C57Bl6 | 47 |
| SA476706 | CC4 | Cecal Contents | C57Bl6 | 48 |
| SA476707 | CC2 | Cecal Contents | C57Bl6 | 49 |
| SA476708 | CC6 | Cecal Contents | C57Bl6 | 60 |
| SA476709 | CC3 | Cecal Contents | C57Bl6 | 62 |
| SA476710 | CC1 | Cecal Contents | C57Bl6 | 63 |
| SA476711 | PV15 | Portal Vein | 129 J | 29 |
| SA476712 | PV14 | Portal Vein | 129 J | 33 |
| SA476713 | PV17 | Portal Vein | 129 J | 34 |
| SA476714 | PV18 | Portal Vein | 129 J | 35 |
| SA476715 | PV16 | Portal Vein | 129 J | 39 |
| SA476716 | PV13 | Portal Vein | 129 J | 42 |
| SA476717 | PV8 | Portal Vein | 129 T | 28 |
| SA476718 | PV10 | Portal Vein | 129 T | 30 |
| SA476719 | PV11 | Portal Vein | 129 T | 31 |
| SA476720 | PV7 | Portal Vein | 129 T | 32 |
| SA476721 | PV12 | Portal Vein | 129 T | 36 |
| SA476722 | PV9 | Portal Vein | 129 T | 37 |
| SA476723 | PV5 | Portal Vein | C57Bl6 | 25 |
| SA476724 | PV4 | Portal Vein | C57Bl6 | 26 |
| SA476725 | PV2 | Portal Vein | C57Bl6 | 27 |
| SA476726 | PV6 | Portal Vein | C57Bl6 | 38 |
| SA476727 | PV3 | Portal Vein | C57Bl6 | 40 |
| SA476728 | PV1 | Portal Vein | C57Bl6 | 41 |
| Showing results 1 to 54 of 54 |
Collection:
| Collection ID: | CO004274 |
| Collection Summary: | At the end of the 12 week HFD diet, animals (n = 6) for each genotype were fasted for 2 hours, anesthetized with Avertin (100 ul per 10 g of BW), after which cecal contents and portal and cardiac blood were collected. All experiments were approved by the IACUC of the Joslin Diabetes Center and were in accordance with NIH guidelines. |
| Sample Type: | Blood (plasma), Cecal contents |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR004290 |
| Treatment Summary: | Animals were started on a high fat diet containing 60% fat by calories (HFD, D12492, Research Diets). All mice continued these diets for 12 weeks. The genotypes tested in this study were C57Bl/6J (B6J) and 129S1 (129J) from Jackson Laboratory (Bar Harbor, ME), and 129S6 (129T) from Taconic Farms (Germantown, NY). B6J mice from Jackson Laboratory are prone to developing obesity and insulin resistance when fed a HFD, whereas 129J and 129T mice are metabolic syndrome resistant. While genetically similar, the inclusion of 129J and 129T mice from different vendors provides a rich microbiome diversity reference of comparison in this study. |
| Animal Endp Euthanasia: | Cardiac Puncture Plasma, Portal Vein Plasma, Cecal Contents |
Sample Preparation:
| Sampleprep ID: | SP004287 |
| Sampleprep Summary: | Snap-frozen cecal samples were thawed on ice, and then aqueous homogenates were generated by homogenizing samples in 10 volumes of water (mg: µL) using a TissueLyser II (QIAGEN) bead mill set to two 2 min intervals at 20Hz. The homogenate for each sample was divided into two 10 µL and two 30 µL aliquots in 1.5mL centrifuge tubes for LC-MS sample preparation LC–MS samples were profiled in each LC-MS method as follows: - HILIC-pos: Cecal or plasma samples (10 μL) were extracted with the addition of nine volumes of 74.9:24.9:0.2 v/v/v acetonitrile/methanol/formic acid containing stable isotope-labeled internal standards (valine-d8, Isotec; and phenylalanine-d8, Cambridge Isotope Laboratories). The samples were centrifuged (10 min, 9,000g, 4°C), and the supernatants (10 μL) injected directly onto column. - C8-pos: Samples (10 μL) were extracted using 190 μL isopropanol containing 1-dodecanoyl-2-tridecanoyl-sn-glycero-3-phosphocholine as an internal standard (Avanti Polar Lipids; Alabaster, AL). After centrifugation (10 min, 9,000g, ambient temperature), supernatants (2 μL) were injected directly onto column. - C18-neg: Samples (30 μL) were extracted using 90 μl methanol containing 15R-15-methyl Prostaglandin A2,15R-15-methyl Prostaglandin F2α, 15S-15-methyl Prostaglandin D2, 15S-15-methyl Prostaglandin E1, and 15S-15-methyl Prostaglandin E2 as internal standards (Cayman Chemical Co.) and centrifuged (10 min, 9,000g, 4°C). The supernatants (10 μL) were injected onto column. - HILIC-neg: Samples (30 μL) were extracted with the addition of four volumes of 80% methanol containing inosine-15N4, thymine-d4 and glycocholate-d4 internal standards (Cambridge Isotope Laboratories). The samples were centrifuged (10 min, 9,000g, 4°C) and the supernatants 10 μL) were injected directly onto column. |
Chromatography:
| Chromatography ID: | CH005202 |
| Instrument Name: | Shimadzu Nexera X2 |
| Column Name: | Water Atlantis HILIC (150 x 2.1 mm x 3 um) |
| Column Temperature: | 30°C |
| Flow Gradient: | Chromatographic separation was performed using an isocratic elution at a flow rate of 250 μL/min with 5% mobile phase A (10 mM ammonium formate and 0.1% formic acid in water) for 1 minute. This was followed by a linear gradient to 40% mobile phase B (acetonitrile with 0.1% formic acid) over 10 minutes. At 10 minutes, the gradient was returned to initial isocratic conditions (5% mobile phase A) and held until 18 minutes, at which point MS acquisition was stopped. The column was equilibrated with 5% mobile phase A at a flow rate of 400 μL/min for 12 minutes, followed by a reduction to the initial flow rate of 250 μL/min for 2 minutes before the next injection. |
| Flow Rate: | 250 µL/min - 400 μL/min |
| Internal Standard: | L-Phenylalanine-d8 (CIL, DLM-372-1), L-Valine-d8 (Sigma, 486027) |
| Solvent A: | 100% Water; 10 mM Ammonium formate; 0.1% Formic acid |
| Solvent B: | 100% Acetonitrile; 0.1% Formic acid |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH005203 |
| Instrument Name: | Shimadzu Nexera X2 |
| Column Name: | Waters Acquity BEH C8 (100 x 2.1mm, 1.7um) |
| Column Temperature: | 40°C |
| Flow Gradient: | The column was initially eluted isocratically at 450 μL/min with 80% mobile phase A for 1 minute. A linear gradient was then applied to reach 80% mobile phase B over 2 minutes, followed by a further gradient to 100% mobile phase B over 7 minutes. The system was held at 100% mobile phase B for 3 minutes before re-equilibrating at 80% mobile phase A for 9 minutes. Data acquisition ended at 15 minutes. |
| Flow Rate: | 450 µL/min |
| Internal Standard: | 12:0-12:0 PC (Avanti 850335C) |
| Solvent A: | 95% Water/5% Methanol; 10 mM Ammonium acetate; 0.1% Acetic acid |
| Solvent B: | 100% Methanol; 0.1% Acetic acid |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH005204 |
| Instrument Name: | Shimadzu Nexera X2 |
| Column Name: | Phenomenex Luna NH2 (150 x 2mm, 5um) |
| Column Temperature: | 30°C |
| Flow Gradient: | Elution began with 10% mobile phase A and 90% mobile phase B, followed by a linear gradient to 100% mobile phase A over 10 minutes. The gradient was then returned to 10% mobile phase A over 2 minutes and held for 13 minutes to equilibrate the column. MS acquisition stopped at 16 minutes. |
| Flow Rate: | 400 µL/min |
| Internal Standard: | Inosine-15N4,Thymine-d4, Glycocholate-d4, CIL NLM-4264-0.01, DLM-2742-0, DLM-1089-1 respectively |
| Solvent A: | 100% water; 20 mM ammonium acetate; 20 mM ammonium hydroxide |
| Solvent B: | 75% acetonitrile/25% methanol; 10 mM ammonium hydroxide |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH005205 |
| Instrument Name: | Shimadzu Nexera X2 |
| Column Name: | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) |
| Column Temperature: | 45°C |
| Flow Gradient: | The column was eluted isocratically at 450 μL/min with 20% mobile phase A for 3 minutes. A linear gradient was then applied to reach 100% mobile phase B over 12 minutes, followed by a 4.5-minute hold at 100% mobile phase B. The column was re-equilibrated with 20% mobile phase A for 7 minutes. MS acquisition stopped at 20 minutes. |
| Flow Rate: | 450 µL/min |
| Internal Standard: | 50ng/mL 15-methyl PGE1, 15-methyl PGF2a, 15-methyl PGA2, 15-methyl PGD2, 15-methyl PGE2 (Cayman) |
| Solvent A: | 100% Water; 0.01% Formic acid |
| Solvent B: | 100% Acetonitrile; 0.01% Acetic acid |
| Chromatography Type: | Reversed phase |
Analysis:
| Analysis ID: | AN006846 |
| Analysis Type: | MS |
| Chromatography ID: | CH005202 |
| Num Factors: | 54 |
| Num Metabolites: | 145 |
| Has Mz: | 1 |
| Has Rt: | 1 |
| Rt Units: | Minutes |
| Results File: | ST004132_AN006846_Results.txt |
| Units: | Abudances |
| Analysis ID: | AN006847 |
| Analysis Type: | MS |
| Chromatography ID: | CH005203 |
| Num Factors: | 54 |
| Num Metabolites: | 227 |
| Has Mz: | 1 |
| Has Rt: | 1 |
| Rt Units: | Minutes |
| Results File: | ST004132_AN006847_Results.txt |
| Units: | Abudances |
| Analysis ID: | AN006848 |
| Analysis Type: | MS |
| Chromatography ID: | CH005204 |
| Num Factors: | 54 |
| Num Metabolites: | 101 |
| Has Mz: | 1 |
| Has Rt: | 1 |
| Rt Units: | Minutes |
| Results File: | ST004132_AN006848_Results.txt |
| Units: | Abudances |
| Analysis ID: | AN006849 |
| Analysis Type: | MS |
| Chromatography ID: | CH005205 |
| Num Factors: | 54 |
| Num Metabolites: | 111 |
| Has Mz: | 1 |
| Has Rt: | 1 |
| Rt Units: | Minutes |
| Results File: | ST004132_AN006849_Results.txt |
| Units: | Abudances |
