Summary of Study ST002575

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.

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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.

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Study IDST002575
Study TitleMetabolomic profiling of PMM2-CDG patient fibroblasts by GC/MS
Study SummaryAbnormal 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 such as sorbitol and mannitol.
Institute
Mayo Clinic
Last NameRadenkovic
First NameSilvia
Address200 2nd Ave SW Rochester MN, USA
Emailradenkovic.silvia@mayo.edu
Phone507(77) 6-6107
Submit Date2023-04-21
Raw Data AvailableYes
Raw Data File Type(s)mzML
Analysis Type DetailGC-MS
Release Date2023-07-18
Release Version1
Silvia Radenkovic Silvia Radenkovic
https://dx.doi.org/10.21228/M81Q6Q
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

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:SU002676
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606
Genotype Strain:WT/PMM2-CDG
Age Or Age Range:5-45
Gender:Male and female

Factors:

Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Genotype Treatment
SA258379Polyol_SR01 (21)PMM2-CDG 10microM epalrestat
SA258380Polyol_SR01 (22)PMM2-CDG 10microM epalrestat
SA258381Polyol_SR03 (9)PMM2-CDG 10microM epalrestat
SA258382Polyol_SR03 (1)PMM2-CDG 10microM epalrestat
SA258383Polyol_SR01 (15)PMM2-CDG 10microM epalrestat
SA258384Polyol_SR02 (21)PMM2-CDG 10microM epalrestat
SA258385Polyol_SR03 (6)PMM2-CDG 10microM epalrestat
SA258386Polyol_SR03 (2)PMM2-CDG 10microM epalrestat
SA258387Polyol_SR03 (5)PMM2-CDG 10microM epalrestat
SA258388Polyol_SR01 (26)PMM2-CDG 10microM epalrestat
SA258389Polyol_SR01 (25)PMM2-CDG 10microM epalrestat
SA258390Polyol_SR04 (1)PMM2-CDG 10microM epalrestat
SA258391Polyol_SR03 (10)PMM2-CDG 10microM epalrestat
SA258392Polyol_SR04 (2)PMM2-CDG 10microM epalrestat
SA258393Polyol_SR04 (6)PMM2-CDG 10microM epalrestat
SA258394Polyol_SR01 (5)PMM2-CDG 10microM epalrestat
SA258395Polyol_SR04 (9)PMM2-CDG 10microM epalrestat
SA258396Polyol_SR01 (1)PMM2-CDG 10microM epalrestat
SA258397Polyol_SR01 (9)PMM2-CDG 10microM epalrestat
SA258398Polyol_SR02 (28)PMM2-CDG 10microM epalrestat
SA258399Polyol_SR04 (5)PMM2-CDG 10microM epalrestat
SA258400Polyol_SR02 (14)PMM2-CDG Glucose
SA258401Polyol_SR02 (15)PMM2-CDG Glucose
SA258402Polyol_SR02 (5)PMM2-CDG Glucose
SA258403Polyol_SR02 (4)PMM2-CDG Glucose
SA258404Polyol_SR02 (3)PMM2-CDG Glucose+Fructose
SA258405Polyol_SR02 (2)PMM2-CDG Glucose+Fructose
SA258406Polyol_SR02 (12)PMM2-CDG Glucose+Fructose
SA258407Polyol_SR02 (11)PMM2-CDG Glucose+Fructose
SA258408Polyol_SR02 (13)PMM2-CDG Glucose+Fructose
SA258409Polyol_SR02 (1)PMM2-CDG Glucose+Fructose
SA258410Polyol_SR02 (30)PMM2-CDG Vehicle
SA258411Polyol_SR02 (23)PMM2-CDG Vehicle
SA258412Polyol_SR02 (29)PMM2-CDG Vehicle
SA258413Polyol_SR03 (7)PMM2-CDG Vehicle
SA258414Polyol_SR04 (4)PMM2-CDG Vehicle
SA258415Polyol_SR04 (7)PMM2-CDG Vehicle
SA258416Polyol_SR04 (8)PMM2-CDG Vehicle
SA258417Polyol_SR04 (10)PMM2-CDG Vehicle
SA258418Polyol_SR04 (3)PMM2-CDG Vehicle
SA258419Polyol_SR03 (12)PMM2-CDG Vehicle
SA258420Polyol_SR03 (4)PMM2-CDG Vehicle
SA258421Polyol_SR03 (8)PMM2-CDG Vehicle
SA258422Polyol_SR03 (11)PMM2-CDG Vehicle
SA258423Polyol_SR03 (3)PMM2-CDG Vehicle
SA258424Polyol_SR02 (22)PMM2-CDG Vehicle
SA258425Polyol_SR01 (24)PMM2-CDG Vehicle
SA258426Polyol_SR01 (27)PMM2-CDG Vehicle
SA258427Polyol_SR01 (23)PMM2-CDG Vehicle
SA258428Polyol_SR01 (10)PMM2-CDG Vehicle
SA258429Polyol_SR01 (2)PMM2-CDG Vehicle
SA258430Polyol_SR01 (6)PMM2-CDG Vehicle
SA258431Polyol_SR01 (28)PMM2-CDG Vehicle
SA258432Polyol_SR01 (16)PMM2-CDG Vehicle
SA258433Polyol_SR03 (17)WT 10microM epalrestat
SA258434Polyol_SR01 (12)WT 10microM epalrestat
SA258435Polyol_SR03 (14)WT 10microM epalrestat
SA258436Polyol_SR03 (13)WT 10microM epalrestat
SA258437Polyol_SR01 (17)WT 10microM epalrestat
SA258438Polyol_SR01 (11)WT 10microM epalrestat
SA258439Polyol_SR01 (7)WT 10microM epalrestat
SA258440Polyol_SR04 (15)WT 10microM epalrestat
SA258441Polyol_SR04 (16)WT 10microM epalrestat
SA258442Polyol_SR04 (12)WT 10microM epalrestat
SA258443Polyol_SR04 (11)WT 10microM epalrestat
SA258444Polyol_SR01 (3)WT 10microM epalrestat
SA258445Polyol_SR01 (18)WT 10microM epalrestat
SA258446Polyol_SR03 (16)WT 10microM epalrestat
SA258447Polyol_SR02 (31)WT 10microM epalrestat
SA258448Polyol_SR02 (25)WT 10microM epalrestat
SA258449Polyol_SR02 (24)WT 10microM epalrestat
SA258450Polyol_SR02 (32)WT 10microM epalrestat
SA258451Polyol_SR02 (9)WT Glucose
SA258452Polyol_SR02 (10)WT Glucose
SA258453Polyol_SR02 (20)WT Glucose
SA258454Polyol_SR02 (19)WT Glucose
SA258455Polyol_SR02 (17)WT Glucose+Fructose
SA258456Polyol_SR02 (18)WT Glucose+Fructose
SA258457Polyol_SR02 (7)WT Glucose+Fructose
SA258458Polyol_SR02 (8)WT Glucose+Fructose
SA258459Polyol_SR02 (16)WT Glucose+Fructose
SA258460Polyol_SR02 (6)WT Glucose+Fructose
SA258461Polyol_SR04 (13)WT Vehicle
SA258462Polyol_SR03 (15)WT Vehicle
SA258463Polyol_SR04 (14)WT Vehicle
SA258464Polyol_SR02 (26)WT Vehicle
SA258465Polyol_SR01 (19)WT Vehicle
SA258466Polyol_SR04 (18)WT Vehicle
SA258467Polyol_SR04 (17)WT Vehicle
SA258468Polyol_SR02 (27)WT Vehicle
SA258469Polyol_SR02 (33)WT Vehicle
SA258470Polyol_SR01 (20)WT Vehicle
SA258471Polyol_SR03 (19)WT Vehicle
SA258472Polyol_SR01 (13)WT Vehicle
SA258473Polyol_SR03 (18)WT Vehicle
SA258474Polyol_SR01 (8)WT Vehicle
SA258475Polyol_SR01 (4)WT Vehicle
SA258476Polyol_SR02 (34)WT Vehicle
SA258477Polyol_SR01 (14)WT Vehicle
Showing results 1 to 99 of 99

Collection:

Collection ID:CO002669
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:TR002688
Treatment Summary:Cells were treated with either 0 or 10microM epalrestat

Sample Preparation:

Sampleprep ID:SP002682
Sampleprep Summary:metabolites were extracted from cells with 2 mL 80 % MeOH, IS and the supernatant was transferred to a fresh 2 mL Eppendorf tube. Standards containing metabolites of interest were prepared simultaneously and dried overnight together with cellular extracts by vacuum centrifugation at 4 °C. The following day, 20 µL of methoxyamine (MOX) (Sigma Aldrich) was added to the samples and they were incubated for 90 min at 37 °C. Next, 60 µL of N, O-bis(trimethylsilyl)trifluoroacetamide (TMS) (Sigma Aldrich) was added to the samples, which were then incubated at 60 °C for 30 min. Samples were kept overnight in a dry and cool place to allow further derivatization with TMS
Processing Storage Conditions:-80℃
Extract Storage:-80℃
Sample Derivatization:MOX, TMS

Combined analysis:

Analysis ID AN004240
Analysis type MS
Chromatography type GC
Chromatography system Agilent 7890A
Column Agilent HP-5ms (30m x 0.25mm, 0.25um)
MS Type EI
MS instrument type Triple quadrupole
MS instrument name Agilent 7000B
Ion Mode NEGATIVE
Units AUC

Chromatography:

Chromatography ID:CH003147
Chromatography Summary:Agilent 7890A GC (Agilent Technologies) coupled with an HP-5 ms 5 % phenyl methyl silox capillary column (30 m, 0.25 mm, 0.25 um, Agilent Technologies)
Instrument Name:Agilent 7890A
Column Name:Agilent HP-5ms (30m x 0.25mm, 0.25um)
Column Temperature:Gradient
Flow Gradient:The temperature gradient applied was as follows: 2 min 100°C, next 175 °C at Δ20 °C/min, followed by an increase to 230 °C at Δ4 °C/min, kept at 230 °C for 3 min, increased to 300 °C at Δ 40°C/min and lastly kept at 300 °C for 5 min. The column was further baked for another 3 min at 325 °C after the gradient.
Flow Rate:NA
Solvent A:N/A
Solvent B:N/A
Chromatography Type:GC

MS:

MS ID:MS003987
Analysis ID:AN004240
Instrument Name:Agilent 7000B
Instrument Type:Triple quadrupole
MS Type:EI
MS Comments:Mass Hunter Workstation software with the Quantitative Analysis Version B.06.00/Build 6.0.388.0 Specific metabolites were identified based on their fragment formula (SIM)/elution time
Ion Mode:NEGATIVE
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