Summary of Study ST002389
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 PR001536. The data can be accessed directly via it's Project DOI: 10.21228/M8512J 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 | ST002389 |
Study Title | Alterations in SHH signal transduction introduce a state of hypometabolism in sporadic Parkinson's disease |
Study Summary | Induced pluripotent stem cells (iPSC) derived from sporadic Parkinson's disease patients and healthy control subjects were used for disease modeling. iPSC were differentiated towards midbrain dopaminergic neurons. For metabolic analysis, midbrain neuronal precursor cells were cultivated in growth medium supplemented with either 1.25 mM [U-13C]-glutamine or 21.25 mM [U-13C]-glucose. Metabolites were extracted and analyzed using GC-MS. The MetaboliteDetector software was used to analyze chromatograms, calculate mass isotopomer distributions (MIDs) and perform relative comparison of metabolite levels. |
Institute | Helmholtz Centre for Environmental Research |
Department | Institute of Developmental Genetics |
Last Name | Schmidt |
First Name | Sebastian |
Address | Ingolstädter Landstraße 1, 85764 Munich, Germany |
sebastian.schmidt@helmholtz-muenchen.de | |
Phone | +4989318743660 |
Submit Date | 2022-12-03 |
Num Groups | 3 |
Total Subjects | 13 |
Raw Data Available | Yes |
Raw Data File Type(s) | bin |
Analysis Type Detail | GC-MS |
Release Date | 2023-10-06 |
Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
Project ID: | PR001536 |
Project DOI: | doi: 10.21228/M8512J |
Project Title: | Alterations in SHH signal transduction introduce a state of hypometabolism in sporadic Parkinson's disease |
Project Type: | Isotopic labeling |
Project Summary: | Sporadic Parkinson’s Disease (sPD) is a progressive neurodegenerative disorder caused by multiple genetic and environmental factors with largely unknown etiology. Prominent pathological culprits include metabolic as well as mitochondrial alterations which have been identified in patients, however, their relevance at different stages of disease progression or their connection remains largely elusive. Here, human iPSCs from late-onset sPD patients were used for disease modeling. Following long-term in vitro cultivation, exclusively neural cells derived from sPD patients developed reduced mitochondrial respiration and glucose consumption reflecting an sPD-specific state of hypometabolism. A multilayered omics analysis based on transcriptomics, proteomics, and metabolomics allowed us to identify the citric acid cycle as being the bottleneck in sPD metabolism. A 13C metabolic flux analysis further unraveled the a-ketoglutarate dehydrogenase complex as being central for a reduced flux through the citric acid cycle. This resulted in a substrate availability problem for the electron transport chain and thus reduced mitochondrial oxygen consumption and ATP production. Notably, these alterations in cellular metabolism were evoked by altered SHH signal transduction due to dysfunctional primary cilia. Upon inhibiting the enhanced SHH signal transduction in sPD, glucose uptake and the activity of the a-ketoglutarate dehydrogenase complex could be restored. Thus, inhibiting overactive SHH signaling may be a potential neuroprotective therapy during the early stages of sPD. |
Institute: | Helmholtz Centre for Environmental Research |
Department: | Institute of Developmental Genetics |
Last Name: | Schmidt |
First Name: | Sebastian |
Address: | Ingolstädter Landstraße 1, 85764 Munich, Germany |
Email: | sebastian.schmidt@helmholtz-muenchen.de |
Phone: | +4989318743660 |
Contributors: | Karsten Hiller, Alexander Heinz |
Subject:
Subject ID: | SU002478 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Male and female |
Cell Biosource Or Supplier: | ForIPS consortium |
Cell Strain Details: | Midbrain neuronal precursor cells differentiated from patient derived induced pluripotent stem cells |
Species Group: | Mammals |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
mb_sample_id | local_sample_id | disease_state | 13C_label |
---|---|---|---|
SA238064 | blank_3_Glc | blank | Glucose |
SA238065 | blank_2_Glc | blank | Glucose |
SA238066 | blank_1_Glc | blank | Glucose |
SA238067 | blank_3_Gln | blank | Glutamine |
SA238068 | blank_1_Gln | blank | Glutamine |
SA238069 | blank_2_Gln | blank | Glutamine |
SA238070 | iG3G-R1-039_2_Glc | ctrl | Glucose |
SA238071 | iG3G-R1-039_3_Glc | ctrl | Glucose |
SA238072 | iG3G-R1-039_1_Glc | ctrl | Glucose |
SA238073 | i1JF-R1-018_3_Glc | ctrl | Glucose |
SA238074 | i1JF-R1-018_1_Glc | ctrl | Glucose |
SA238075 | i1E4-R1-003_2_Glc | ctrl | Glucose |
SA238076 | i1E4-R1-003_1_Glc | ctrl | Glucose |
SA238077 | i1JF-R1-018_2_Glc | ctrl | Glucose |
SA238078 | iO3H-R1-005_2_Glc | ctrl | Glucose |
SA238079 | i1E4-R1-003_3_Glc | ctrl | Glucose |
SA238080 | i82A-R1-002_3_Glc | ctrl | Glucose |
SA238081 | iO3H-R1-005_3_Glc | ctrl | Glucose |
SA238082 | iO3H-R1-005_1_Glc | ctrl | Glucose |
SA238083 | i82A-R1-002_2_Glc | ctrl | Glucose |
SA238084 | i82A-R1-002_1_Glc | ctrl | Glucose |
SA238085 | i1E4-R1-003_1_Gln | ctrl | Glutamine |
SA238086 | iG3G-R1-039_3_Gln | ctrl | Glutamine |
SA238087 | i1JF-R1-018_2_Gln | ctrl | Glutamine |
SA238088 | i1E4-R1-003_2_Gln | ctrl | Glutamine |
SA238089 | i1JF-R1-018_3_Gln | ctrl | Glutamine |
SA238090 | iG3G-R1-039_2_Gln | ctrl | Glutamine |
SA238091 | iO3H-R1-005_3_Gln | ctrl | Glutamine |
SA238092 | i82A-R1-002_3_Gln | ctrl | Glutamine |
SA238093 | i1JF-R1-018_1_Gln | ctrl | Glutamine |
SA238094 | i82A-R1-002_2_Gln | ctrl | Glutamine |
SA238095 | i82A-R1-002_1_Gln | ctrl | Glutamine |
SA238096 | iO3H-R1-005_1_Gln | ctrl | Glutamine |
SA238097 | iO3H-R1-005_2_Gln | ctrl | Glutamine |
SA238098 | i1E4-R1-003_3_Gln | ctrl | Glutamine |
SA238099 | iG3G-R1-039_1_Gln | ctrl | Glutamine |
SA238100 | iJ2C-R1-015_3_Glc | sPD | Glucose |
SA238101 | iJ2C-R1-015_2_Glc | sPD | Glucose |
SA238102 | iJ2C-R1-015_1_Glc | sPD | Glucose |
SA238103 | iC99-R1-007_3_Glc | sPD | Glucose |
SA238104 | iM89-R1-005_1_Glc | sPD | Glucose |
SA238105 | iM89-R1-005_2_Glc | sPD | Glucose |
SA238106 | iC99-R1-007_1_Glc | sPD | Glucose |
SA238107 | i88H-R1-002_3_Glc | sPD | Glucose |
SA238108 | iM89-R1-005_3_Glc | sPD | Glucose |
SA238109 | iR66-R1-007_1_Glc | sPD | Glucose |
SA238110 | iC99-R1-007_2_Glc | sPD | Glucose |
SA238111 | iPX7-R1-001_2_Glc | sPD | Glucose |
SA238112 | iR66-R1-007_2_Glc | sPD | Glucose |
SA238113 | i88H-R1-002_1_Glc | sPD | Glucose |
SA238114 | i88H-R1-002_2_Glc | sPD | Glucose |
SA238115 | iPX7-R1-001_1_Glc | sPD | Glucose |
SA238116 | iPX7-R1-001_3_Glc | sPD | Glucose |
SA238117 | iR66-R1-007_3_Glc | sPD | Glucose |
SA238118 | iAY6-R1-003_3_Glc | sPD | Glucose |
SA238119 | iAY6-R1-003_1_Glc | sPD | Glucose |
SA238120 | iAY6-R1-003_2_Glc | sPD | Glucose |
SA238121 | iPX7-R1-001_2_Gln | sPD | Glutamine |
SA238122 | iPX7-R1-001_1_Gln | sPD | Glutamine |
SA238123 | iPX7-R1-001_3_Gln | sPD | Glutamine |
SA238124 | i88H-R1-002_3_Gln | sPD | Glutamine |
SA238125 | iAY6-R1-003_3_Gln | sPD | Glutamine |
SA238126 | i88H-R1-002_2_Gln | sPD | Glutamine |
SA238127 | i88H-R1-002_1_Gln | sPD | Glutamine |
SA238128 | iC99-R1-007_1_Gln | sPD | Glutamine |
SA238129 | iM89-R1-005_1_Gln | sPD | Glutamine |
SA238130 | iM89-R1-005_2_Gln | sPD | Glutamine |
SA238131 | iJ2C-R1-015_3_Gln | sPD | Glutamine |
SA238132 | iJ2C-R1-015_2_Gln | sPD | Glutamine |
SA238133 | iJ2C-R1-015_1_Gln | sPD | Glutamine |
SA238134 | iM89-R1-005_3_Gln | sPD | Glutamine |
SA238135 | iC99-R1-007_2_Gln | sPD | Glutamine |
SA238136 | iR66-R1-007_3_Gln | sPD | Glutamine |
SA238137 | iAY6-R1-003_1_Gln | sPD | Glutamine |
SA238138 | iR66-R1-007_2_Gln | sPD | Glutamine |
SA238139 | iR66-R1-007_1_Gln | sPD | Glutamine |
SA238140 | iC99-R1-007_3_Gln | sPD | Glutamine |
SA238141 | iAY6-R1-003_2_Gln | sPD | Glutamine |
Showing results 1 to 78 of 78 |
Collection:
Collection ID: | CO002471 |
Collection Summary: | Neuronal precursor cells differentiated from patient derived induced pluripotent stem cells were seeded on Geltrex coated 6-well plates containing neuronal precursor maintenance medium at a density of 1,000,000 cells/well with six replicates per cell line. hNPCs were cultured at 37 °C, 7 % CO2, 21 % O2 for at least 72 h. |
Sample Type: | Neurons |
Storage Conditions: | -80? |
Treatment:
Treatment ID: | TR002490 |
Treatment Summary: | Growth medium was replaced by a labeling medium containing the respective stable-isotope tracer instead of its unlabeled variant. Cells were cultured with either 1.25 mM [U-13C]-glutamine or 21.25 mM [U-13C]-glucose for 24 h. |
Sample Preparation:
Sampleprep ID: | SP002484 |
Sampleprep Summary: | Subsequently, cell culture supernatant was stored for profiling the extracellular metabolome. Three replicates per cell line and blanks were washed with 0.9% NaCl and quenched with ice-cold methanol and ice-cold ddH2O (containing 1 µg/ml D6-glutaric acid as internal standard). Cells were scraped and extracts were added into tubes containing ice-cold chloroform. Following vortexing at 1,400 rpm for 20 min at 4°C and centrifugation at 17,000 g for 5 min at 4°C, 300 µl of the polar phase were transferred into GC glass vials with microinsert and dried under vacuum at 4°C. Dried extracts were derivatized using equal amounts of methoxylamine (20 mg/ml in pyridine) and MTBSTFA. |
Combined analysis:
Analysis ID | AN003893 |
---|---|
Analysis type | MS |
Chromatography type | GC |
Chromatography system | Agilent 7890B gas chromatograph |
Column | 30 m DB-35 ms and 5 m Duruguard capillary column |
MS Type | EI |
MS instrument type | Single quadrupole |
MS instrument name | Agilent 5977 |
Ion Mode | POSITIVE |
Units | Mass isotopomer distributions |
Chromatography:
Chromatography ID: | CH002883 |
Instrument Name: | Agilent 7890B gas chromatograph |
Column Name: | 30 m DB-35 ms and 5 m Duruguard capillary column |
Chromatography Type: | GC |
MS:
MS ID: | MS003633 |
Analysis ID: | AN003893 |
Instrument Name: | Agilent 5977 |
Instrument Type: | Single quadrupole |
MS Type: | EI |
MS Comments: | One microliter sample was injected into an SSL injector at 270 °C in a split or splitless mode. GC-MS analysis was performed using an Agilent 7890B gas chromatograph equipped with a 30 m DB-35 ms and 5 m Duruguard capillary column. Metabolites were detected in selected ion mode by an Agilent 5977 MSD system. The MetaboliteDetector software was used to analyze chromatograms, calculate mass isotopomer distributions (MIDs) and perform relative comparison of metabolite levels. |
Ion Mode: | POSITIVE |