Summary of Study ST003108
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 PR001931. The data can be accessed directly via it's Project DOI: 10.21228/M84428 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 | ST003108 |
| Study Title | Complete absence of GLUT1 does not impair human terminal erythroid differentiation |
| Study Summary | The Glucose transporter 1 (GLUT1) is one of the most abundant proteins within the erythrocyte membrane and is required for glucose and dehydroascorbic acid (Vitamin C precursor) transport. It is widely recognized as a key protein for red cell structure, function, and metabolism. Previous reports highlighted the importance of GLUT1 activity within these uniquely glycolysis-dependent cells, in particular for increasing antioxidant capacity needed to avoid irreversible damage from oxidative stress in humans. However, studies of glucose transporter roles in erythroid cells are complicated by species-specific differences between humans and mice. Here, using CRISPRmediated gene editing of immortalized erythroblasts and adult CD34+ hematopoietic progenitor cells, we generate committed human erythroid cells completely deficient in expression of GLUT1. We show that absence of GLUT1 does not impede human erythroblast proliferation, differentiation, or enucleation. This work demonstrates for the first-time generation of enucleated human reticulocytes lacking GLUT1. The GLUT1-deficient reticulocytes possess no tangible alterations to membrane composition or deformability in reticulocytes. Metabolomic analyses of GLUT1-deficient reticulocytes reveal hallmarks of reduced glucose import, downregulated metabolic processes and upregulated AMPK-signalling, alongside alterations in antioxidant metabolism, resulting in increased osmotic fragility and metabolic shifts indicative of higher oxidant stress. Despite detectable metabolic changes in GLUT1 deficient reticulocytes, the absence of developmental phenotype, detectable proteomic compensation or impaired deformability comprehensively alters our understanding of the role of GLUT1 in red blood cell structure, function and metabolism. It also provides cell biological evidence supporting clinical consensus that reduced GLUT1 expression does not cause anaemia in GLUT1 deficiency syndrome. |
| Institute | University of Colorado |
| Last Name | Stephenson |
| First Name | Daniel |
| Address | Research 1 South L18-1303 12801 E. 17th Ave., Aurora, Colorado, 80045, USA |
| daniel.stephenson@cuanschutz.edu | |
| Phone | 303-724-3339 |
| Submit Date | 2024-02-27 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzXML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-03-20 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN005087 | AN005088 | AN005089 | AN005090 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish | Thermo Vanquish | Thermo Vanquish |
| Column | Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) | Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) | Phenomenex Kinetex C18 (30 x 2.1mm, 1.7um) | Phenomenex Kinetex C18 (30 x 2.1mm, 1.7um) |
| MS Type | ESI | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE | POSITIVE | NEGATIVE |
| Units | area | area | area | area |
MS:
| MS ID: | MS004824 |
| Analysis ID: | AN005087 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | El-Maven used for data analysis. Orbitrap scanning in full MS mode from 65-975 m/z at 60,000 resolution, with 50 Arb sheath gas, 10 Arb auxiliary gas, and 3.4 kV spray voltage. |
| Ion Mode: | POSITIVE |
| MS ID: | MS004825 |
| Analysis ID: | AN005088 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | El-Maven used for data analysis. Orbitrap scanning in full MS mode from 65-975 m/z at 60,000 resolution, with 50 Arb sheath gas, 10 Arb auxiliary gas, and 3 kV spray voltage. |
| Ion Mode: | NEGATIVE |
| MS ID: | MS004826 |
| Analysis ID: | AN005089 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Lipidsearch used for data analysis. Orbitrap scanning using data dependent MS2 (top 10) from 125-1500 m/z at 17,500 resolution, with 45 Arb sheath gas, 25 Arb auxiliary gas, and 4 kV spray voltage. |
| Ion Mode: | POSITIVE |
| MS ID: | MS004827 |
| Analysis ID: | AN005090 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Lipidsearch used for data analysis. Orbitrap scanning using data dependent MS2 (top 10) from 125-1500 m/z at 17,500 resolution, with 45 Arb sheath gas, 25 Arb auxiliary gas, and 4 kV spray voltage. |
| Ion Mode: | NEGATIVE |
