Summary of Study ST002503
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 PR001617. The data can be accessed directly via it's Project DOI: 10.21228/M8PH89 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 | ST002503 |
| Study Title | Endothelial Cell CD36 Regulates Membrane Ceramide Formation, Exosome Fatty Acid Delivery to Tissues and Circulating Fatty Acid Levels |
| Study Type | Membrane ceramide and Fatty Acid Uptake |
| Study Summary | Endothelial cell (EC) CD36 controls tissue fatty acid (FA) uptake. Here we visualized FA transfer by ECs using click chemistry. Apical FA interaction with EC CD36 induces actin reorganization, caveolin-1 tyrosine 14 (Cav-1Y14) phosphorylation, and ceramide generation in caveolae, culminating in caveolae internalization into FA/CD36/ceramide vesicles that are secreted basolaterally as small extracellular vesicles, sEVs (exosomes). In transwells, ECs expressing fluorescent CD63 (exosomal marker) transfer FAs to underlying myotubes in CD63-labeled sEVs. In vivo FA-CD63-CD36 transfer from ECs to muscle is shown using a mouse with EC-specific expression of emGFP-CD63. CD36 depletion, actin-binding latrunculin-B, Src inhibition, Cav-1Y14 mutation, and membrane neutral sphingomyelinase inhibition by GW4869 helped map the FA-sEV pathway. Injecting GW4869 to mice reduced muscle FA uptake, raised plasma FAs and lowered glucose, mimicking prominent Cd36-/- phenotypes. The FA-sEV pathway we describe contributes to crosstalk between ECs and underlying cells and links regulation of membrane ceramide to blood FAs. |
| Institute | Washington University in St. Louis |
| Department | IM-Nutitrional Science |
| Laboratory | Abumrad Lab |
| Last Name | Palacios |
| First Name | Hector |
| Address | West Building 00201, St. Louis, Missouri, 63110, USA |
| hectorp@wustl.edu | |
| Phone | 314-362-5397 |
| Submit Date | 2023-03-06 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-03-22 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN004112 | AN004113 | AN004114 |
|---|---|---|---|
| Analysis type | MS | MS | MS |
| Chromatography type | Reversed phase | HILIC | HILIC |
| Chromatography system | Agilent 1290 Infinity II | Thermo Vanquish | Thermo Vanquish |
| Column | Waters CORTECS UPLC C18 (150 x 2.1mm,1.6um) | Waters CORTECS HILIC (100 x 2.1mm,1.6um) | Waters CORTECS HILIC (100 x 2.1mm,1.6um) |
| MS Type | ESI | ESI | ESI |
| MS instrument type | QTOF | Orbitrap | Orbitrap |
| MS instrument name | Agilent 6545 QTOF | Thermo Orbitrap ID-X tribrid | Thermo Orbitrap ID-X tribrid |
| Ion Mode | POSITIVE | POSITIVE | NEGATIVE |
| Units | Peak Area | Peak Area | Peak Area |
Chromatography:
| Chromatography ID: | CH003047 |
| Methods Filename: | LC_Method.pdf |
| Instrument Name: | Agilent 1290 Infinity II |
| Column Name: | Waters CORTECS UPLC C18 (150 x 2.1mm,1.6um) |
| Column Temperature: | 60 |
| Flow Gradient: | 0-2 min, 30% B; 17 min, 75% B; 20 min, 85% B; 23-26 min, 100% B; 26 min, 30% B followed by a re-equilibration phase of 5 min. |
| Flow Rate: | .25ml/min |
| Solvent A: | A: 60% acetonitrile, 40% water, 0.1% formic acid, 10 mM ammonium formate, 2.5 mM medronic acid |
| Solvent B: | 90% 2-propanol, 10% acetonitrile, 0.1% formic acid, 10 mM ammonium formate |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH003048 |
| Methods Filename: | LC_Method.pdf |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters CORTECS HILIC (100 x 2.1mm,1.6um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0 to 1 min, 90% B; 12 min, 35% B; 12.5 to 14.5 min, 25% B; 15 min, 90% B followed by a re-equilibration phase of 4 |
| Flow Rate: | .25ml/min |
| Solvent A: | A: 95% water, 5% acetonitrile, 20 mM ammonium bicarbonate, 0.1% ammonium hydroxide solution (25% ammonia in water), 2.5 μM medronic acid |
| Solvent B: | B: 95% acetonitrile, 5% water, 2.5 μM medronic acid |
| Chromatography Type: | HILIC |
