New Perspectives in Dried Blood Spot Biomarkers for Lysosomal Storage Diseases

Abstract

Dried blood spots (DBSs) biomarkers are convenient for monitoring for specific lysosomal storage diseases (LSDs), but they could have relevance for other LSDs. To determine the specificity and utility of glycosphingolipidoses biomarkers against other LSDs, we applied a multiplexed lipid liquid chromatography tandem mass spectrometry assay to a DBS cohort of healthy controls (n = 10) and Gaucher (n = 4), Fabry (n = 10), Pompe (n = 2), mucopolysaccharidosis types I–VI (n = 52), and Niemann–Pick disease type C (NPC) (n = 5) patients. We observed no complete disease specificity for any of the markers tested. However, comparison among the different LSDs highlighted new applications and perspectives of the existing biomarkers. We observed elevations in glucosylceramide isoforms in the NPC and Gaucher patients relative to the controls. In NPC, there was a greater proportion of C24 isoforms, giving a specificity of 96–97% for NPC, higher than 92% for the NPC biomarker N-palmitoyl-O-phosphocholineserine ratio to lyso-sphingomyelin. We also observed significantly elevated levels of lyso-dihexosylceramide in Gaucher and Fabry disease as well as elevated lyso-globotriaosylceramide (Lyso-Gb3) in Gaucher disease and the neuronopathic forms of Mucopolysaccharidoses. In conclusion, DBS glucosylceramide isoform profiling has increased the specificity for the detection of NPC, thereby improving diagnostic accuracy. Low levels of lyso-lipids can be observed in other LSDs, which may have implications in their disease pathogenesis.

Modeling Lysosomal Storage Disorders in an Innovative Way: Establishment and Characterization of Stem Cell Lines from Human Exfoliated Deciduous Teeth of Mucopolysaccharidosis Type II Patients

“…However, disruptions of the glycosphingolipid degradation pathways are implicated not only in glycophingolipidoses but also in other LSDs, including MPSs [ 15 ]. And it is true that not only there is a well-documented secondary accumulation of lipids in MPS diseases (MPS II included) [ 21 , 58 ] but also there is an increasing body of evidence documenting significant elevations of several glycophingolipidose biomarkers in other LSDs, including neuronopathic forms of MPSs [ 59 ]. That is why we tried to address the question of whether any of those biomarkers could be altered in the cell lines under analysis.…”

Application of tandem mass spectrometry in the screening and diagnosis of mucopolysaccharidoses

“…Neurofibrillary light chain is significantly elevated in cerebrospinal fluid (CSF) and serum of MPS II patients with neurological involvement, and they are positively correlated with CSF HS, which can be used to assess neuronal health and damage in treated MPS II patients [ 8 ]. Lysozyme B3 has been found to be a potential secondary biomarker of nervous system involvement in MPS patients [ 6 , 72 ]. Elevated expression of three candidate biomarkers, matrix metalloproteinase 19, α-trypsin interinhibitor heavy chain 3, and α-1-microglobulin, was demonstrated in MPS I cartilage, and serum abundance of these molecules was found to correlate with MRI and histologic degeneration grading [ 92 ].…”

“…Although disease-specific biomarker levels can overlap with other similar diseases, the use of multiple biomarkers can help establish a diagnosis more rapidly. Examination of a range of pathophysiologically relevant markers could also help to provide insight into common downstream pathophysiologic mechanisms leading to intracellular dysfunction in these diseases [ 72 ]. Based on current findings, GAGs and their types remain the most commonly used biomarkers for MPSs.…”

The Liver and Lysosomal Storage Diseases: From Pathophysiology to Clinical Presentation, Diagnostics, and Treatment

“…The use of dried blood spots (DBSs) in the diagnostics process of LSDs has become increasingly popular, mainly due to its convenience. In addition to enzyme activity measurement, simultaneous diagnostics based on substrate identification is possible, i.e., analysis of glucosphingosine (lyso-Gb1) concentration in GD and lyso-sphingomyelin (lyso-SPM) in ASMD [ 50 , 51 , 52 ]. Once NPC is suspected clinically, several metabolites (cholestane-3β, 5α, 6β-triol, lyso-sphingomyelin isoforms—lyso-SM and lyso-SM509—and bile acid metabolites) have emerged as sensitive and specific diagnostic biomarkers for NPC and their study, completed by genetic analyses ( NPC1 and NPC2 genes), and they should now be considered as the first line in laboratory testing [ 53 ].…”