Abstract
Introduction
Pompe disease is a rare, lysosomal disorder, characterized by intra-lysosomal glycogen accumulation due to an impaired function of α-glucosidase enzyme. The laboratory testing for Pompe is usually performed by enzyme activity, genetic test, or urine glucose tetrasaccharide (Glc4) screening by HPLC. Despite being a good preliminary marker, the Glc4 is not specific for Pompe.
Objective
The purpose of the present study was to develop a simple methodology using liquid chromatography-high resolution mass spectrometry (LC-HRMS) for targeted quantitative analysis of Glc4 combined with untargeted metabolic profiling in a single analytical run to search for complementary biomarkers in Pompe disease.
Methods
We collected 21 urine specimens from 13 Pompe disease patients and compared their metabolic signatures with 21 control specimens.
Results
Multivariate statistical analyses on the untargeted profiling data revealed Glc4, creatine, sorbitol/mannitol, L-phenylalanine, N-acetyl-4-aminobutanal, N-acetyl-L-aspartic acid, and 2-aminobenzoic acid as significantly altered in Pompe disease. This panel of metabolites increased sample class prediction (Pompe disease versus control) compared with a single biomarker.
Conclusion
This study has demonstrated the potential of combined acquisition methods in LC-HRMS for Pompe disease investigation, allowing for routine determination of an established biomarker and discovery of complementary candidate biomarkers that may increase diagnostic accuracy, or improve the risk stratification of patients with disparate clinical phenotypes.
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Data availability
Fig. S1-S3 and Tables S1-S4 are provided as supplementary material. The raw LC-HRMS files can be found at the MassIVE Repository (Dataset MSV000088329).
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This work was supported by Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro-FAPERJ (process E26/010.002501/2019) and Coordenação de Aperfeiçoamento de Nível Superior-CAPES (Finance code 001).
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Conception and design: MBMM, HMRS, MLCO, FBS, RWAP, and RG; Formal analysis and investigation: MBMM and HMRS; Writing - Original Draft: HMRS, RWAP, and RG; Writing: - Review and Editing: MLCO, FBS, RWAP, and RG; Resources: MLCO, FBS, and RG; Supervision and corresponding author: RG.
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de Moraes, M.B.M., de Souza, H.M.R., de Oliveira, M.L.C. et al. Combined targeted and untargeted high-resolution mass spectrometry analyses to investigate metabolic alterations in pompe disease. Metabolomics 19, 29 (2023). https://doi.org/10.1007/s11306-023-01989-w
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DOI: https://doi.org/10.1007/s11306-023-01989-w