Diagnosis of sphingolipidoses: a new simultaneous measurement of lysosphingolipids by LC-MS/MS

Polo, Giulia; Burlina, Alessandro P.; Kolamunnage, Thilini B.; Zampieri, Michele; Dionisi‐Vici, Carlo; Strisciuglio, Pietro; Zaninotto, Martina; Plebani, Mario; Burlina, Alberto

doi:10.1515/cclm-2016-0340

Cited by 92 publications

(94 citation statements)

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“…The GlcSph level was not analyzed separately for technical reasons. Separate measurement of GlcSph and GalSph by LC–MS/MS has been performed previously by Polo and colleagues . HexSph species increases in GD patients were qualitatively confirmed to be ClcSph, and in increases in Krabbe disease patients, they were GalSph.…”

Section: Discussionmentioning

confidence: 87%

“…We adopted the LC‐MS/MS method for simultaneous quantification of the following several LysoSLs: HexSph (GalSph+GlcSph), LysoGb3, and LysoSM, including the new biomarker LysoSM‐509, in DBS. This method was developed for rapid and effective biochemical diagnostics for several sphingolipidoses and applied for LysoSLs quantification in blood plasma . The ceramide metabolic pathway is shown in Figure .…”

Section: Methodsmentioning

confidence: 99%

“…DBS (HexSph [GlcSph + GalSph], LysoGb3, LysoSM, and LysoSM509) levels were measured by the LC–MS/MS method as described previously with modifications (Table ). During chromatographic separation, GlcSph and its isomer GalSph elute in a single peak named as HexSph.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, lysosphingolipids (LysoSLs) have been identified as storage compounds in several sphingolipidoses, including Gaucher, Fabry, and Niemann‐Pick diseases. A method of simultaneous LysoSL quantification by liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) has been developed . It is worth noting that GlcSph is the most prevalent plasma sphingolipid compared to GlcCer and is believed to be the most sensitive biomarker of GD …”

mentioning

confidence: 99%

“…A method of simultaneous LysoSL quantification by liquid chromatographytandem mass spectrometry (LC-MS/MS) has been developed. 8 It is worth noting that GlcSph is the most prevalent plasma sphingolipid compared to GlcCer and is believed to be the most sensitive biomarker of GD. [5][6][7] The link between GBA mutations and PD remains unknown.…”

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confidence: 99%

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Blood lysosphingolipids accumulation in patients with parkinson's disease with glucocerebrosidase 1 mutations

et al. 2018

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Section: Discussionmentioning

confidence: 87%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Blood lysosphingolipids accumulation in patients with parkinson's disease with glucocerebrosidase 1 mutations

et al. 2018

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Lysosomal Storage Diseases. For Better or Worse: Adapting to Defective Lysosomal Glycosphingolipid Breakdown

Aerts

Ferraz

Mirzaian

et al. 2017

Encyclopedia of Life Sciences

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The cellular recycling of glycosphingolipids (GSLs) is mediated by specific lysosomal glycosidases. Inherited deficiencies in these enzymes cause lysosomal storage disorders. Some of the common disorders are Gaucher disease (GD) and Fabry disease (FD) resulting from the defects in lysosomal glucocerebrosidase (GBA) degrading glucosylceramide and α‐galactosidase A (GLA) degrading globotriaosylceramide. Here, GSL accumulation in tissues slows down with age despite ongoing lysosomal turnover of endogenous and endocytosed GSLs. Biochemical adaptations might explain this phenomenon. One crucial adaptation is the deacylation of accumulating GSLs in lysosomes by acid ceramidase. The soluble bases glucosylsphingosine in GD and globotriaosylsphingosine in FD are capable of leaving lysosomes and cells. In the case of GD, a further adaptation involves the cytosol‐faced enzyme GBA2. This enzyme allows extra‐lysosomal degradation of GlcCer while possibly generating glucosylated cholesterol. The beneficial and harmful effects of these adaptations are discussed. Key Concepts Glycosphingolipids (GSLs) are membrane constituents composed of a ceramide with one or more sugars. The simplest GSL is glucosylceramide (GlcCer). Ongoing recycling of GSLs in cells includes lysosomal degradation by the sequential action of glycosidases and acid ceramidase. Deficiency of lysosomal glycosidase leads to lysosomal storage diseases caused by accumulation of the corresponding substrate in lysosomes. The most common glycosphingolipidoses are Gaucher disease (GD) and Fabry disease (FD). GD is an autosomal recessive disorder caused by deficient activity of the lysosomal enzyme acid β‐glucosidase (glucocerebrosidase; GBA) resulting in lysosomal accumulation of GlcCer. FD is an X‐linked disorder caused by deficient activity of the lysosomal enzyme α‐galactosidase A (GLA) resulting in lysosomal accumulation of globotriaosylceramide (Gb3). Accumulation of storage lipids during GBA and GLA tends to slow down with age, likely partly due to poorly appreciated biochemical adaptations. Active conversion of accumulating GlcCer in lysosomes of GBA‐deficient cells is mediated by acid ceramidase, resulting in the formation of water‐soluble glucosylsphingosine (GlcSph). Likewise, globotriaosylsphingosine (lysoGb3) is formed from accumulating in lysosomes of GLA‐deficient cells. Elevated plasma GlcSph and lysoGb3 levels can be sensitively measured LC–MS and may assist in diagnosing and monitoring of the disease and response to treatment in GD and FD patients, respectively. Increased GlcSph level in GD patients acts as an autoantigen, causing ongoing B‐cell proliferation, leading to multiple myeloma. Increased lysoGb3 level in FD patients is thought to cause damage to nociceptive neurons and podocytes, thus contributing to pain and renal failure. In GD, the cytosol‐faced enzyme β‐glucosidase GBA2 allows degradation of GlcCer outside lysosomes. Through transglycosylation, GBA2 may generate glucosylcholesterol and ceramide from GlcCer and cholesterol. The toxic effects of secondary metabolites such as glycosphingoid bases (GlcSph in GD and lysoGb3 in FD) and glucosylated metabolites (GlcChol in GD) warrant further investigations.

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Lysosphingolipid Quantitation in Plasma and Dried‐Blood Spots Using Targeted High‐Resolution Mass Spectrometry

Ducatez,

Mauhin,

Ottaviani

et al. 2024

Clinical Laboratory Analysis

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BackgroundSphingolipidoses are rare inherited metabolic diseases belonging to lysosomal diseases. Early and accurate diagnosis is crucial for effective management and treatment. In this study, we aimed to develop a robust method to accelerate the diagnosis of these sphingolipidoses using dried blood spots and plasma.MethodWe employed high‐resolution mass spectrometry coupled with liquid chromatography (LC‐HRMS) to analyze 6 lysosphingolipids (GlcSph/Psychosine, LysoGb3, LysoSM, LysoSM509, LysoGM1, and LysoGM2) on dried blood spots and plasma samples. The method was used to measure the lysosphingolipid levels in a group of 30 control subjects and 204 samples from patients with sphingolipidoses (61 dB and 143 plasma) including Fabry, Gaucher, GM2 Gangliodosis, Niemann‐Pick type A/B, and Niemann‐Pick type C.ResultsThe developed multiplex LC‐HRMS method demonstrated linearity, precision, and quantification performances particularly for GlcSph/Psychosine and LysoGb3 on samples including controls and patients with sphingolipidoses. LysoSM showed recovery variability, wherease LysoGM1 and LysoGM2 showed higher matrix effect.ConclusionOur study presents a high‐resolution mass spectrometry method along with the established cutoff values, providing a valuable tool for targeted screening, accurate diagnosis, and monitoring sphingolipidoses. Furthermore, DBS showed reliable results that lay the path to a broader adoption for screening these diseases.

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Diagnosis of sphingolipidoses: a new simultaneous measurement of lysosphingolipids by LC-MS/MS

Cited by 92 publications

References 26 publications

Blood lysosphingolipids accumulation in patients with parkinson's disease with glucocerebrosidase 1 mutations

Blood lysosphingolipids accumulation in patients with parkinson's disease with glucocerebrosidase 1 mutations

Lysosomal Storage Diseases. For Better or Worse: Adapting to Defective Lysosomal Glycosphingolipid Breakdown

Lysosphingolipid Quantitation in Plasma and Dried‐Blood Spots Using Targeted High‐Resolution Mass Spectrometry

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