Quantitation of a Urinary Profile of Biomarkers in Gaucher Disease Type 1 Patients Using Tandem Mass Spectrometry
Iskren Menkovic,
Michel Boutin,
Abdulfatah Alayoubi,
Filipa Curado,
Peter Bauer,
François E. Mercier,
Christiane Auray-Blais
Affiliations
Iskren Menkovic
Division of Medical Genetics, Department of Pediatrics, Centre de Recherche-CHUS, Faculty of Medicine and Health Sciences, Université de Sherbrooke, CIUSSS de l’Estrie-CHUS, 3001, 12th Avenue North, Sherbrooke, QC J1H 5N4, Canada
Michel Boutin
Division of Medical Genetics, Department of Pediatrics, Centre de Recherche-CHUS, Faculty of Medicine and Health Sciences, Université de Sherbrooke, CIUSSS de l’Estrie-CHUS, 3001, 12th Avenue North, Sherbrooke, QC J1H 5N4, Canada
Abdulfatah Alayoubi
Divisions of Experimental Medicine and Hematology, Department of Medicine, Faculty of Medicine, McGill University, Lady Davis Institute for Medical Research, Jewish General Hospital, 3755, Côte Sainte-Catherine, Montreal, QC H3T 1E2, Canada
Filipa Curado
CENTOGENE GmbH, 18055 Rostock, Germany
Peter Bauer
CENTOGENE GmbH, 18055 Rostock, Germany
François E. Mercier
Divisions of Experimental Medicine and Hematology, Department of Medicine, Faculty of Medicine, McGill University, Lady Davis Institute for Medical Research, Jewish General Hospital, 3755, Côte Sainte-Catherine, Montreal, QC H3T 1E2, Canada
Christiane Auray-Blais
Division of Medical Genetics, Department of Pediatrics, Centre de Recherche-CHUS, Faculty of Medicine and Health Sciences, Université de Sherbrooke, CIUSSS de l’Estrie-CHUS, 3001, 12th Avenue North, Sherbrooke, QC J1H 5N4, Canada
Gaucher disease is a rare inherited disorder caused by a deficiency of the lysosomal acid beta-glucocerebrosidase enzyme. Metabolomic studies by our group targeted several new potential urinary biomarkers. Apart from lyso-Gb1, these studies highlighted lyso-Gb1 analogs −28, −26, −12 (A/B), +2, +14, +16 (A/B), +30, and +32 Da, and polycyclic lyso-Gb1 analogs 362, 366, 390, and 394 Da. The main objective of the current study was to develop and validate a robust UPLC-MS/MS method to study the urine distribution of these biomarkers in patients. Method: Urine samples were purified using solid-phase extraction. A 12 min UPLC-MS/MS method was developed. Results: Validation assays revealed high precision and accuracy for creatinine and lyso-Gb1. Most lyso-Gb1 analogs had good recovery rates and high intra- and interday precision assays. Biomarker-estimated LOD and LOQ levels ranged from 56–109 pM to 186–354 pM, respectively. Comparison between GD patients and healthy controls showed significant differences in most biomarker levels. Typically, treated GD patients presented lower biomarker levels compared to untreated patients. Conclusions: These data suggest that the metabolites investigated might be interesting GD biomarkers. More studies with a larger cohort of patients will be needed to better understand the clinical significance of these GD biomarkers.
