Quantitation of a Plasma Biomarker Profile for the Early Detection of Gaucher Disease type 1 Patients

Abstract

Aim: Gaucher disease (GD) is caused by a deficiency of the lysosomal enzyme acid β-glucocerebrosidase. Recent metabolomic studies highlighted several new metabolites increased in the plasma of GD patients. We aimed to develop and validate a UPLC–MS/MS method allowing a relative quantitation of lyso-Gb1 and lyso-Gb1 analogs -28, -12, -2, +14, +16 and +18 Da in addition to sphingosylphosphorylcholine, N-palmitoyl- O-phosphocholine to study potential correlations with clinical manifestations. Methodology & results: Following solid-phase extraction, plasma samples were evaporated and resuspended in 100 μl of resuspension solution. Three microliter is injected into the UPLC–MS/MS for analysis. Conclusion: All biomarkers studied were increased in GD patients. Significant correlations were observed between specific analogs and hematological, and visceral complications, as well as overall disease severity.

Multiplex Quantification of Plasma Biomarkers for Patients with Gaucher Disease Type 1

“…This is a major departure from the previous dogma, which held that GD biomarkers are only useful for patient monitoring and follow‐up. We performed a semi‐targeted metabolomic study of plasma specimens obtained from patients with untreated GD and healthy controls (Menkovic et al., 2022a) based on our previous experience with Fabry disease (Auray‐Blais et al., 2012; Dupont, Gagnon, Boutin, and Auray‐Blais, 2013). Based on the results of this study and the research done by Mirzaian and colleagues (Mirzaian et al., 2015), we developed and validated a UPLC‐MS/MS method to quantify lyso‐Gb 1 and lyso‐Gb 1 analogs at –28 Da, –12 Da, –2 Da, +14 Da, +16 Da, and +18 Da in addition to N ‐palmitoyl‐O‐phosphocholineserine (NPOPCS) and sphingosylphosphorylcholine (SPC) to study the distribution of these markers in treated and untreated patients (Menkovic et al., 2022b).…”

“…This is a major departure from the previous dogma, which held that GD biomarkers are only useful for patient monitoring and follow-up. We performed a semi-targeted metabolomic study of plasma specimens obtained from patients with untreated GD and healthy controls (Menkovic et al, 2022a) based on our previous experience with Fabry disease (Auray-Blais et al, 2012;Dupont, Gagnon, Boutin, and Auray-Blais, 2013). Based on the results of this study…”

“…This specific metabolomic study using ultra‐performance liquid chromatography time‐of‐flight mass spectrometry highlighted several potential biomarkers, namely lyso‐Gb 1 and four related analogs (with the following modifications on the sphingosine moiety: ‐C 2 H 4 (‐28 Da), ‐H 2 (‐2 Da), ‐H 2 +O (+14 Da), and +H 2 O (+18 Da)), sphingosylphosphorylcholine (SPC), and N‐ palmitoyl‐O‐phosphocholineserine (NPOPCS) (Menkovic et al., 2020). After structural identification of these metabolites, we developed and validated a quantification method for these compounds and two other lyso‐Gb 1 analogs (see Background Information) using ultra‐performance liquid chromatography/tandem mass spectrometry (UPLC‐MS/MS) to investigate marker distributions and their clinical significance (Menkovic et al., 2022a). The validation step revealed that the method had high sensitivity with low limits of quantification and detection as well as high levels of accuracy and precision (Menkovic et al., 2022a).…”

“…After structural identification of these metabolites, we developed and validated a quantification method for these compounds and two other lyso‐Gb 1 analogs (see Background Information) using ultra‐performance liquid chromatography/tandem mass spectrometry (UPLC‐MS/MS) to investigate marker distributions and their clinical significance (Menkovic et al., 2022a). The validation step revealed that the method had high sensitivity with low limits of quantification and detection as well as high levels of accuracy and precision (Menkovic et al., 2022a). Herein, we provide an in depth protocol for the method developed to study the distribution of metabolites identified in our metabolomic study.…”