Summary of Study ST002934
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 PR001825. The data can be accessed directly via it's Project DOI: 10.21228/M8T714 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 | ST002934 |
| Study Title | Metabolic profiling of newborn DBS samples associated with transit and false elevation of glutarylcarnitine (C5DC) |
| Study Summary | Background: Glutaric aciduria type-1 (GA-1) is a rare autosomal recessive metabolic disorder caused by a glutaryl coenzyme A dehydrogenase (GCDH) deficiency, affecting approximately 1 in 110,000 individuals globally. This enzymatic deficiency leads to abnormal elevations of glutaryl-CoA and its derivatives, specifically glutaric acid (GA), 3-hydroxyglutaric acid (3OHGA), and glutarylcarnitine (C5DC). Clinical manifestations encompass macrocephaly, developmental delays, and movement disorders. Early detection via genetic testing and newborn screening (NBS), utilizing GA-1 biomarkers in dried blood spot (DBS) samples, is vital for prompt intervention. Despite the NBS system, transit-elevated C5DC-containing DBS samples from falsely suspected GA-1 newborns sometimes yield normal results, posing diagnostic sensitivity and specificity challenges. Consequently, there is a growing need for alternative diagnostic tools. Comprehensive mass spectrometry-based untargeted metabolomics offers promise in identifying additional informative biomarkers for distinguishing falsely suspected GA-1 newborns from healthy counterparts. Methodology: In this prospective study, we obtained DBS samples with transit-elevated C5DC levels from falsely suspected GA-1 newborns (n=47) and matched control DBS samples from healthy newborns (n=47) through the NBS program. Metabolites were extracted and analyzed via liquid chromatography-high-resolution mass spectrometry (LC-HRMS). Subsequent multivariate and univariate statistical analyses and feature annotation enabled biomarker and pathway investigations for significantly altered metabolites. Results: Untargeted metabolomics analysis revealed alterations in 582 upregulated and 546 downregulated metabolites. The commonly used GA-1 biomarkers, including C5DC, exhibited no significant changes in the falsely suspected GA-1 DBS samples. Conversely, 155 endogenous metabolites displayed significant variations compared to the control group. Furthermore, our data identified novel altered metabolic biomarkers, such as N-Palmitoylcysteine, 3-hydroxylinoleoylcarnitine, Heptacarboxyporphyrin, and MG (0:0/20:1/0:0), along with perturbed metabolic pathways like sphingolipid and thiamine metabolism associated with the transient and falsely elevated C5DC levels in DBS samples. Conclusions: Our untargeted metabolomics investigation unveiled distinct metabolic pathways and biomarkers linked to the transient C5DC elevation in DBS samples from falsely suspected GA-1 newborns. These findings can enhance GA-1 diagnosis by serving as predictive indicators during NBS analysis. Validation studies are warranted to confirm the presence of these newly identified metabolic pathways and biomarkers in confirmed GA-1 cases. |
| Institute | King Faisal Specialist Hospital and Research Centre (KFSHRC) |
| Last Name | Al Mogren |
| First Name | Maha |
| Address | Zahrawi Street, Al Maather, Riyadh 11211, Saudi Arabia |
| mahamogren@gmail.com | |
| Phone | 966541205332 |
| Submit Date | 2023-09-11 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Waters) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-04-02 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN004812 | AN004813 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Waters Acquity | Waters Acquity |
| Column | Waters Acquity UPLC XSelect HSS C18 (100 × 2.1mm, 2.5um) | Waters Acquity UPLC XSelect HSS C18 (100 × 2.1mm, 2.5um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Waters Xevo-G2-S | Waters Xevo-G2-S |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Peak Area | Peak Area |
Chromatography:
| Chromatography ID: | CH003637 |
| Chromatography Summary: | The Waters Acquity UPLC system coupled with a Xevo G2-S QTOF mass spectrometer equipped with an electrospray ionization source (ESI) was used to analyze the samples. The extracted metabolites were chromatographed using an ACQUITY UPLC using XSelect (100×2.1mm, 2.5 μm) column (Waters Ltd., Elstree, UK), the mobile phase composed of 0.1% formic acid in dH2O as solvent A, and B consists of 0.1% formic acid in 50% ACN: MeOH. A gradient elution schedule was run as follows: 0-16 min 95- 5% A, 16-19 min 5% A, 19-20 min 5-95% A, 20-22 min 95- 95% A, at 300 µL/min flow rate. MS spectra were acquired under positive and negative electrospray ionization modes (ESI+, ESI-). MS conditions were as follows: source temperature was 150◦C, the desolvation temperature was 500◦C (ESI+) or 140 (ESI−), the capillary voltage was 3.20 kV (ESI+) or 3 kV (ESI−), cone voltage was 40 V, desolvation gas flow was 800.0 L/h, cone gas flow was 50 L/h. The collision energies of low and high functions were set at off and 10 V to 50 V, respectively, in MSE mode. The mass spectrometer was calibrated with 100–1200 Da sodium formate in both ionization modes. The lock mass compound, leucine-enkephalin (an external reference to the ion m/z 556.2771 in (ESI+) and 554.2615 (ESI-)), was infused continuously, switching between the sample and the reference every 45 and 60 s for ESI+ and ESI-, respectively, for a 0.5 s scan time, a flow rate of 10 µL/min, a cone voltage of 30 V, and collision energy of 4 V. Data were collected in continuum mode with Masslynx™ V4.1 workstation (Waters Inc., Milford, Massachusetts, USA). The samples were acquired randomly, and a QC sample was injected after each 10 study samples to control the system’s variations. A mixture of several internal standards with a final concentration of 10 µg/ml was added to each sample, serving as a reference compound to help correct any variations that may occur during sample preparation and analysis. |
| Methods Filename: | GA_LCMS Metabolomics |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters Acquity UPLC XSelect HSS C18 (100 × 2.1mm, 2.5um) |
| Column Temperature: | 55 |
| Flow Gradient: | 0–16 min 95%–5% A, 16–19 min 5% A, 19–20 min 5%–95% A, and 20–22 min, 95%– 95% A |
| Flow Rate: | 300 μl/min. |
| Solvent A: | 0.1% formic acid in dH2O |
| Solvent B: | 0.1% formic acid in 50% MeOH and ACN |
| Chromatography Type: | Reversed phase |
