MSACL 2016 EU Abstract

Mass Spectrometry Applications: from Biomarker Discovery to Detection of Patients with Fabry Disease Having a Late-Onset Cardiac Mutation

Christiane Auray-Blais (Presenter)
Université de Sherbrooke

Bio: Christiane Auray-Blais is Professor in the Faculty of Medicine and Health Sciences at the Université de Sherbrooke, Quebec, Canada. She also holds positions as Scientific Director at Waters-CHUS Expertise Centre in Clinical Mass Spectrometry at the Clinical Research Center-CHUS and Director of the Quebec Neonatal Urine Screening Program. After receiving her Bachelor’s degree in biochemistry, Master’s degree in Health Law, and PhD in radiology from the Université de Sherbrooke, Dr Auray-Blais went on to complete post-doctoral studies at Duke University, North Carolina, USA. Dr Auray-Blais’ research interests include biomarkers and screening, particularly in the area of inborn errors of metabolism. She has expertise in the use of mass spectrometry for biomarker discovery and method development for screening, diagnosis, monitoring and follow up of patients. She has won numerous awards for her research work.

Authorship: Christiane Auray-Blais (1), Pamela Lavoie (1), Michel Boutin (1), Huang Chun-Kai (2), Hsu Ting-Rong (2,3), Dau-Ming Niu (2,3)
(1) Université de Sherbrooke, Sherbrooke, QC Canada; (2) Taipei Veterans General Hospital, Taipei, Taiwan; (3) National Yang-Ming University, Taipei, Taiwan

Short Abstract

A time-of-flight mass spectrometry metabolomic approach led to the discovery of novel biomarkers for Fabry disease, a lysosomal storage disorder. The discovery of urine (n=7) and plasma (n=6) analogues of lyso-Gb3 is particularly useful for the detection of Fabry patients having late-onset cardiac mutations. Incidence in Taiwan: 1:1600. Tandem mass spectrometry methods were devised to analyze these biomarkers in a cohort of 191 Fabry patients with a cardiac mutation. Results show an association with some analogues of lyso-Gb3 and the left ventricular mass index and the Mainz Severity Score Index, thus an efficient clinical application for detecting these patients.

Long Abstract

Rationale: Fabry disease is an X-linked, panethnic lysosomal storage disorder caused by a deficiency of á-galactosidase A leading to accumulation of glycosphingolipids in various organs, biological fluids and the vascular endothelium (1). The prevalence of the Fabry disease mutation IVS4 + 919G>A (IVS4) associated with the late-onset cardiac phenotype in the Taiwanese population was estimated at 1:1600 males and 1:800 females through newborn screening (2). These patients have high residual enzyme activity and they present symptoms, such as hypertrophic cardiomyopathy, renal failure, or cryptogenic stroke at later stages in life. Usual biomarkers for Fabry disease, such as the globotriaosylceramide (Gb3), were mostly normal in these patients, thus delaying their detection and treatment (3). Lyso-Gb3 (m/z = 786) analogues were detected by a time-of-flight metabolomic approach (4) and tandem mass spectrometry methods were devised and validated in urine and plasma (5,6). These analogues of lyso-Gb3 present modifications on the sphingosine moiety of the molecule.

Objectives: The objective of this research project was to investigate the relationship between globotriaosylceramide (Gb3), globotriaosylsphingosine (lyso-Gb3) and related analogues and clinical manifestations of the disease in a large cohort of patients having a cardiac phenotype.

Design and Methods: Urinary and plasma lyso-Gb3 and related analogues, and urinary Gb3 were analyzed using tandem mass spectrometry in a cohort of 191 Fabry patients carrying the IVS4 mutation. Of these 191 individuals, 79 were under 18 years of age (pediatric group) and 112 were adults. Statistical analyses were achieved using IBM SPSS Statistics version 22. Multiple regression analyses were performed to determine the association between biomarker levels and left ventricular mass index (LVMI) and Mainz Severity Score Index (MSSI), adjusted for gender and age.

Results: In males (n=25), a positive correlation was found between the following urinary analogues of lyso-Gb3 biomarkers and age: lyso-Gb3 (+16) (rs=0.726, p<0.01), lyso-Gb3 (+34) (rs=0.795, p<0.01), and lyso-Gb3 (+50) (rs=0.673, p<0.01). In males (n=22), a positive correlation was also found between plasma lyso-Gb3 and age (rs=0.596, p<0.01). The majority of male patients older than 40 years of age were symptomatic, and globally had higher levels of these specific biomarkers. As expected with an X-linked disorder, our results show that females were generally less clinically affected than males with Fabry disease and females had lower levels of biomarkers compared to males. Moreover, our results show that the plasma level of lyso-Gb3, and urine analogue levels of lyso-Gb3 at (+16), (+34), and (+50) had a positive association with the LVMI, and/or the MSSI. Furthermore, children showing high levels of these biomarkers were also identified. Interestingly, some of these patients had family members who were more severely affected than other Fabry patients in the same age range.

Conclusions: Lyso-Gb3 and related analogues might be reliable biomarkers for patients with the IVS4 mutation, considering the observed association with LVMI and MSSI in the cohort under study. Longitudinal biomarker follow-up of children with the IVS4 mutation would be important to determine if the elevation of urinary and plasma Fabry biomarkers at a young age are predictive of the clinical outcomes of the disease, as well as its severity in adulthood.

References & Acknowledgements:

Acknowledgements: This research was funded by a grant-in-aid of research from Shire. We are grateful to Waters Corporation for their continued scientific support and partnership. We would like to thank all Fabry patients for their participation in this study.


1. Clarke, JTR. Narrative review: Fabry disease. Ann Intern Med. 2007;146:425−433.

2. Lin HY, Chong KW, Hsu JH, et al. High incidence of the cardiac variant of Fabry disease revealed by newborn screening in the Taiwan Chinese population. Circ Cardiovasc Genet. 2009;2(5):450-456.

3. Auray-Blais C, Blais CM, Ramaswami U, Boutin M, Germain DP, Dyack S, Bodamer O, Pintos-Morell G, Clarke JTR, Bichet DG, Warnock DG, Echevarria L, West ML, Lavoie P. Urinary biomarker investigation in children with Fabry disease using tandem mass spectrometry. Clin Chim Acta. 2015;438:195-204.

4. Auray-Blais C, Boutin M, Gagnon R, Dupont FO, Lavoie P, Clarke JT. Urinary globotriaosylsphingosine-related biomarkers for Fabry disease targeted by metabolomics. Anal Chem. 2012;84(6):2745-2753.

5. Lavoie P, Boutin M, Auray-Blais C. Multiplex analysis of novel urinary lyso-Gb3-related biomarkers for Fabry disease by tandem mass spectrometry. Anal Chem. 2013;85(3):1743-1752.

6. Boutin M, Auray-Blais C. Multiplex tandem mass spectrometry analysis of novel plasma lyso-Gb3-related analogues in Fabry disease. Anal Chem. 2014;86(7):3476-3483.

Financial Disclosure

GrantsyesShire, Genzyme, A Sanofi Division, BioMarin, Waters
Board Memberno
ExpensesyesShire, Genzyme, A Sanofi Division, BioMarin, Shire, Genzyme, A Sanofi Division, BioMarin, Waters

IP Royalty: no

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