Summary of Study ST002565
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 PR001653. The data can be accessed directly via it's Project DOI: 10.21228/M81Q6Q 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 | ST002565 |
| Study Title | Metabolomic profiling of PMM2-CDG after siRNA mediated KD of AKR1b1 - 13C6 glucose and fructose study |
| Study Summary | Abnormal polyol metabolism has been predominantly associated with diabetes, where excess glucose is converted to sorbitol by aldose reductase (AR). Recently, abnormal polyol metabolism has also been implicated in phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), and epalrestat, an AR inhibitor, proposed as a potential therapy for this disorder. Given that the PMM enzyme is not closely connected to polyol metabolism, and, unlike in diabetes, PMM2-CDG does not present with hyperglycemia in blood, the increased polyol production, and the therapeutic mechanism of epalrestat in PMM2-CDG remained largely elusive. PMM2-CDG is caused by deficiency of the PMM enzyme and results in a depletion of mannose-1-P and guanosine diphosphate mannose (GDP-mannose), which is essential for glycosylation. Here, we show that apart from glycosylation abnormalities, PMM2 deficiency also leads to changes in intracellular glucose flux, which results in an increase in intracellular polyols. Ssing tracer glucose studies, we demonstrate that AR inhibition diverts glucose flux away from polyol production towards the synthesis of sugar nucleotides. |
| Institute | Mayo Clinic |
| Last Name | Radenkovic |
| First Name | Silvia |
| Address | 200 2nd Ave SW Rochester MN, USA |
| radenkovic.silvia@mayo.edu | |
| Phone | 507(77) 6-6107 |
| Submit Date | 2023-04-18 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-07-18 |
| Release Version | 1 |
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Project:
| Project ID: | PR001653 |
| Project DOI: | doi: 10.21228/M81Q6Q |
| Project Title: | Metabolomic profiling of PMM2-CDG zebrafish in presence and absence of epalrestat |
| Project Summary: | Abnormal polyol metabolism has been predominantly associated with diabetes, where excess glucose is converted to sorbitol by aldose reductase (AR). Recently, abnormal polyol metabolism has also been implicated in phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), and epalrestat, an AR inhibitor, proposed as a potential therapy for this disorder. Given that the PMM enzyme is not closely connected to polyol metabolism, and, unlike in diabetes, PMM2-CDG does not present with hyperglycemia in blood, the increased polyol production, and the therapeutic mechanism of epalrestat in PMM2-CDG remained largely elusive. PMM2-CDG is caused by deficiency of the PMM enzyme and results in a depletion of mannose-1-P and guanosine diphosphate mannose (GDP-mannose), which is essential for glycosylation. Here, we show that apart from glycosylation abnormalities, PMM2 deficiency also leads to changes in intracellular glucose flux, which results in an increase in intracellular polyols. Targeting AR with epalrestat decreases polyol levels and increases GDP-mannose in vivo in pmm2 mutant zebrafish. |
| Institute: | Mayo Clinic |
| Last Name: | Radenkovic |
| First Name: | Silvia |
| Address: | 200 2nd Ave SW Rochester MN, USA |
| Email: | radenkovic.silvia@mayo.edu |
| Phone: | 507(77) 6-6107 |
| Funding Source: | NIH, KU Leuven |
| Publications: | Tracer metabolomics reveals the role of aldose reductase in glycosylation |
| Contributors: | Silvia Radenkovic, Anna N. Ligezka, Sneha S. Mokashi, Karen Driesen, Lynn Dukes-Rimsky, Graeme Preston, Luckio F. Owuocha, Leila Sabbagh, Jehan Mousa, Christina Lam, Andrew Edmondson, Austin Larson, Matthew Schultz, Pieter Vermeersch, David Cassiman, Peter Witters, Lesa J. Beamer, Tamas Kozicz, Heather Flanagan-Steet, Bart Ghesquière, Eva Morava |
Subject:
| Subject ID: | SU002666 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Genotype Strain: | WT/PMM2-CDG |
| Age Or Age Range: | 40-45 |
| Gender: | Female |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Genotype | Treatment |
|---|---|---|---|
| SA257927 | SR08 | PMM2-CDG | 12C fructose + 12C glucose |
| SA257928 | SR04 | PMM2-CDG | 12C fructose + 12C glucose |
| SA257929 | SR03 | PMM2-CDG | 12C fructose + 12C glucose |
| SA257930 | SR07 | PMM2-CDG | 12C fructose + 12C glucose |
| SA257931 | SR05 | PMM2-CDG | 12C fructose+13C fructose |
| SA257932 | SR06 | PMM2-CDG | 12C fructose+13C fructose |
| SA257933 | SR02 | PMM2-CDG | 13C glu + 12C fructose |
| SA257934 | SR01 | PMM2-CDG | 13C glu + 12C fructose |
| SA257935 | SR10 | PMM2-CDG | 5.5mM 13C glucose |
| SA257936 | SR09 | PMM2-CDG | 5.5mM 13C glucose |
| SA257937 | SR11 | PMM2-CDG | 5.5mM glucose + |
| SA257938 | SR12 | PMM2-CDG | 5.5mM glucose + |
| SA257939 | SR19 | WT | 12C fructose + 12C glucose |
| SA257940 | SR16 | WT | 12C fructose + 12C glucose |
| SA257941 | SR15 | WT | 12C fructose + 12C glucose |
| SA257942 | SR20 | WT | 12C fructose + 12C glucose |
| SA257943 | SR18 | WT | 12C fructose+13C fructose |
| SA257944 | SR17 | WT | 12C fructose+13C fructose |
| SA257945 | SR13 | WT | 13C glu + 12C fructose |
| SA257946 | SR14 | WT | 13C glu + 12C fructose |
| SA257947 | SR24 | WT | 5.5mM 13C glucose |
| SA257948 | SR23 | WT | 5.5mM 13C glucose |
| SA257949 | SR22 | WT | 5.5mM glucose + |
| SA257950 | SR21 | WT | 5.5mM glucose + |
| Showing results 1 to 24 of 24 |
Collection:
| Collection ID: | CO002659 |
| Collection Summary: | Cells were washed with PBS, cells were incubated with extraction buffer for 2min before scraping and transferring to a fresh eppendorf. Samples were precipitated overnight at -80, then they were centrifuged at max rpm, 20min, 4 degrees C and supernatant transferred to an M/S vial. |
| Sample Type: | Fibroblasts |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002678 |
| Treatment Summary: | Cells were treated with 12C/13C glucose +/- 12C/13C fructose |
Sample Preparation:
| Sampleprep ID: | SP002672 |
| Sampleprep Summary: | Metabolites were extracted with 80% Methanol and IS (d27 myristic acid) |
| Processing Storage Conditions: | -80℃ |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN004226 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters Acquity |
| Column | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | NEGATIVE |
| Units | AUC |
Chromatography:
| Chromatography ID: | CH003135 |
| Chromatography Summary: | C18 iP REVERSE PHASE |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) |
| Column Temperature: | 40 |
| Flow Gradient: | The gradient started with 5% of solvent B and 95% solvent A and remained at 5% B until 2 min post injection. A linear gradient to 37% B was carried out until 7 min and increased to 41% until 14 min. Between 14 and 26 minutes the gradient increased to 95% of B and remained at 95% B for 4 minutes. At 30 min the gradient returned to 5% B. The chromatography was stopped at 40 min. |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | 100% water; 10mM tributylamine; 15mM acetic acid |
| Solvent B: | 100% methanol |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003973 |
| Analysis ID: | AN004226 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | El-Maven polly, ThermoFisher Xcalibur; Metabolites were annotated using the inhouse standard metabolite library- elution time and m/z values. |
| Ion Mode: | NEGATIVE |
