Summary of Study ST004399
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 PR002783. The data can be accessed directly via it's Project DOI: 10.21228/M8XK1H 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 | ST004399 |
| Study Title | Intracellular polar metabolomics on neural progenitor cells (NPCs) generated using dual SMAD inhibition method from Fragile X Syndrome (FXS) and CRISPR edited isogenic rescue induced pluripotent stem cells (iPSCs). |
| Study Summary | To test FMRP dependence of metabolite changes, we examined NPCs generated from two independent FXS lines and their CRISPR-edited isogenic rescue counterparts and did polar metabolomics on them. LC/MS profiling of polar metabolites (triplicates per line) showed enrichment of increased intracellular metabolites in both rescue lines relative to their FXS counterparts. A clear genotypic segregation was also observed. |
| Institute | UMass Chan Medical School |
| Last Name | UMass Chan |
| First Name | Spinelli Lab |
| Address | 55 Lake Avenue North, Worcester, Massachusetts, 01605, USA |
| spinellilab@gmail.com | |
| Phone | (508) 856-8989 ext. 68148 |
| Submit Date | 2025-11-13 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-12-01 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002783 |
| Project DOI: | doi: 10.21228/M8XK1H |
| Project Title: | Metabolic Reprogramming during Human Neuron Differentiation Indicates Glutaminase as a Key Determinant in Fragile X Syndrome |
| Project Summary: | Metabolic homeostasis gone awry is a contributor to, if not an underlying cause of, several neurologic disorders. Fragile X syndrome (FXS) is a neurodevelopmental disorder caused by a trinucleotide repeat expansion in FMR1 and consequent loss of the encoded protein FMRP, which results in downstream molecular, neurologic, and mitochondrial deficits that are linked to cognitive impairment. In human postmortem brain, many metabolites and solute carrier proteins are coordinately dysregulated, which also occurs during differentiation of human iPSCs into excitatory neurons. Metabolic tracing in FXS neurons demonstrates a dearth of glutamine deamidation to glutamate, which reduces anaplerosis into the TCA cycle, potentially hindering bioenergetic and biosynthetic functions of mitochondria. Mechanistically, aberrant expression of glutaminase isoforms in FXS is responsible for reduced glutaminolysis, thereby altering glutamate levels which may contribute to FXS. |
| Institute: | UMass Chan Medical School |
| Last Name: | UMass Chan |
| First Name: | Richter Lab |
| Address: | 55 Lake Avenue North, Worcester, Massachusetts, 01605, USA |
| Email: | spinellilab@gmail.com |
| Phone: | (508) 856-8989 ext. 68148 |
Subject:
| Subject ID: | SU004558 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Genotype |
|---|---|---|---|
| SA521228 | FXS 1 | Male Homo sapien CRISPR rescue NPC | FXS |
| SA521229 | FXS 2 | Male Homo sapien CRISPR rescue NPC | FXS |
| SA521230 | FXS 3 | Male Homo sapien CRISPR rescue NPC | FXS |
| SA521231 | FXS-SW 1 | Male Homo sapien CRISPR rescue NPC | FXS |
| SA521232 | FXS-SW 2 | Male Homo sapien CRISPR rescue NPC | FXS |
| SA521233 | FXS-SW 3 | Male Homo sapien CRISPR rescue NPC | FXS |
| SA521234 | iControl/E3 1 | Male Homo sapien CRISPR rescue NPC | TD |
| SA521235 | iControl/E3 2 | Male Homo sapien CRISPR rescue NPC | TD |
| SA521236 | iControl/E3 3 | Male Homo sapien CRISPR rescue NPC | TD |
| SA521237 | C1-2 1 | Male Homo sapien CRISPR rescue NPC | TD |
| SA521238 | C1-2 2 | Male Homo sapien CRISPR rescue NPC | TD |
| SA521239 | C1-2 3 | Male Homo sapien CRISPR rescue NPC | TD |
| Showing results 1 to 12 of 12 |
Collection:
| Collection ID: | CO004551 |
| Collection Summary: | iPSC lines were cultured on Matrigel coated plates in mTeSR™1 (Catalog #85850) with 10uM ROCK inhibitor (Y-27632). Neural induction was performed using the embryoid body formation protocol from StemCell technologies. Briefly cells were plated on Matrigel coated AggreWell™800 wells in STEMdiff™ Neural Induction Medium + SMADi and embryoid bodies were allowed to form for 13 days followed by Rosette selection as per manufactures instructions. The rosettes were plated on Poly-L-Ornithine (PLO)/ laminin coated plates for 3 days and then dissociated into single cells and expanded as NPCs in STEMdiff™ Neural Progenitor Medium. |
| Sample Type: | Cultured cells |
Treatment:
| Treatment ID: | TR004567 |
| Treatment Summary: | To generate neuron precursors, NPCs were cultured in STEMdiff™ Forebrain Neuron Differentiation media (Catalog #08600) for 3 days on PLO/laminin plates followed by replating at a density of 135,000 cells/cm2 in PLO/laminin plates in STEMdiff™ Forebrain Neuron Maturation media (Catalog #08605) with Compound E for 4 weeks. |
Sample Preparation:
| Sampleprep ID: | SP004564 |
| Sampleprep Summary: | For extraction from cell pellets from in-vitro cultures, samples were incubated on dry ice with cold 80% methanol. The extracts were spun down and the supernatant was vacuum dried before resuspending in LC/MS grade water. |
Chromatography:
| Chromatography ID: | CH005575 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Merck SeQuant ZIC-HILIC (150 x 2.1mm,5um) |
| Column Temperature: | 25 |
| Flow Gradient: | 20 min, 80% - 20% B; 0.5 min, 20% - 80% B; 7.5min, 80% B |
| Flow Rate: | 0.15ml/min |
| Solvent A: | 100% water; 0.1% ammonium hydroxide; 20mM ammonium carbonate |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
Analysis:
| Analysis ID: | AN007353 |
| Analysis Type: | MS |
| Chromatography ID: | CH005575 |
| Num Factors: | 2 |
| Num Metabolites: | 92 |
| Units: | Peak Area |
| Analysis ID: | AN007354 |
| Analysis Type: | MS |
| Chromatography ID: | CH005575 |
| Num Factors: | 2 |
| Num Metabolites: | 139 |
| Units: | Peak Area |
