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Plasma lysosphingomyelin demonstrates great potential as a diagnostic biomarker for Niemann-Pick disease type C in a retrospective study.

Welford RW, Garzotti M, Marques Lourenço C, Mengel E, Marquardt T, Reunert J, Amraoui Y, Kolb SA, Morand O, Groenen P - PLoS ONE (2014)

Bottom Line: Lysosphingolipids have been shown to be excellent biomarkers of sphingolipidosis in several enzyme deficient lysosomal storage disorders.Plasma SPC and GlcSph are stable and can be measured accurately, precisely and reproducibly.Plasma GlcSph did not correlate with SPC levels in NP-C patients.

View Article: PubMed Central - PubMed

Affiliation: Actelion Pharmaceuticals Ltd, Allschwil, Switzerland.

ABSTRACT
Niemann-Pick disease type C (NP-C) is a devastating, neurovisceral lysosomal storage disorder which is characterised by variable manifestation of visceral signs, progressive neuropsychiatric deterioration and premature death, caused by mutations in the NPC1 and NPC2 genes. Due to the complexity of diagnosis and the availability of an approved therapy in the EU, improved detection of NP-C may have a huge impact on future disease management. At the cellular level dysfunction or deficiency of either the NPC1 or NPC2 protein leads to a complex intracellular endosomal/lysosomal trafficking defect, and organ specific patterns of sphingolipid accumulation. Lysosphingolipids have been shown to be excellent biomarkers of sphingolipidosis in several enzyme deficient lysosomal storage disorders. Additionally, in a recent study the lysosphingolipids, lysosphingomyelin (SPC) and glucosylsphingosine (GlcSph), appeared to be elevated in the plasma of three adult NP-C patients. In order to investigate the clinical utility of SPC and GlcSph as diagnostic markers, an in-depth fit for purpose biomarker assay validation for measurement of these biomarkers in plasma by liquid chromatography-tandem mass spectrometry was performed. Plasma SPC and GlcSph are stable and can be measured accurately, precisely and reproducibly. In a retrospective analysis of 57 NP-C patients and 70 control subjects, median plasma SPC and GlcSph were significantly elevated in NP-C by 2.8-fold and 1.4-fold respectively. For miglustat-naïve NP-C patients, aged 2-50 years, the area under the ROC curve was 0.999 for SPC and 0.776 for GlcSph. Plasma GlcSph did not correlate with SPC levels in NP-C patients. The data indicate excellent potential for the use of lysosphingomyelin in NP-C diagnosis, where it could be used to identify NP-C patients for confirmatory genetic testing.

No MeSH data available.


Related in: MedlinePlus

Glucosylsphingosine not galactosylsphingosine is increased in the plasma of NP-C patients.LC-MS/MS chromatograms showing separation of glucosyl- and galactosyl-sphingosine with HILIC chromatography. (A) QC2; (B) QC2 spiked with 5 nM glucosylsphingosine; (C) QC2 spiked with 5 nM galactosylsphingosine; (D) control sample 1; (E) control sample 2; (F) control sample 3; (G) NP-C sample 1; (H) NP-C sample 2; (I) NP-C sample 3.
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pone-0114669-g006: Glucosylsphingosine not galactosylsphingosine is increased in the plasma of NP-C patients.LC-MS/MS chromatograms showing separation of glucosyl- and galactosyl-sphingosine with HILIC chromatography. (A) QC2; (B) QC2 spiked with 5 nM glucosylsphingosine; (C) QC2 spiked with 5 nM galactosylsphingosine; (D) control sample 1; (E) control sample 2; (F) control sample 3; (G) NP-C sample 1; (H) NP-C sample 2; (I) NP-C sample 3.

Mentions: Subsequent to the main study a sub-study was designed to investigate if the hexosylsphingosine peak corresponded to glucosylsphingosine (GlcSph) or galactosylsphingosine (GalSph). To achieve separation of GlcSph and GalSph it was necessary to switch to a HILIC stationary phase for the chromatography so that interactions were dominated by the polar sugar moiety. GlcSph was found to elute before GalSph (Figure 6). In the control samples there was ∼3-fold more GlcSph than GalSph. In the three NP-C patient samples, the increase above normal levels was dominated by GlcSph, leading to an increase in the GlcSph/GalSph ratio (to >12) (Figure 6).


Plasma lysosphingomyelin demonstrates great potential as a diagnostic biomarker for Niemann-Pick disease type C in a retrospective study.

Welford RW, Garzotti M, Marques Lourenço C, Mengel E, Marquardt T, Reunert J, Amraoui Y, Kolb SA, Morand O, Groenen P - PLoS ONE (2014)

Glucosylsphingosine not galactosylsphingosine is increased in the plasma of NP-C patients.LC-MS/MS chromatograms showing separation of glucosyl- and galactosyl-sphingosine with HILIC chromatography. (A) QC2; (B) QC2 spiked with 5 nM glucosylsphingosine; (C) QC2 spiked with 5 nM galactosylsphingosine; (D) control sample 1; (E) control sample 2; (F) control sample 3; (G) NP-C sample 1; (H) NP-C sample 2; (I) NP-C sample 3.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4257710&req=5

pone-0114669-g006: Glucosylsphingosine not galactosylsphingosine is increased in the plasma of NP-C patients.LC-MS/MS chromatograms showing separation of glucosyl- and galactosyl-sphingosine with HILIC chromatography. (A) QC2; (B) QC2 spiked with 5 nM glucosylsphingosine; (C) QC2 spiked with 5 nM galactosylsphingosine; (D) control sample 1; (E) control sample 2; (F) control sample 3; (G) NP-C sample 1; (H) NP-C sample 2; (I) NP-C sample 3.
Mentions: Subsequent to the main study a sub-study was designed to investigate if the hexosylsphingosine peak corresponded to glucosylsphingosine (GlcSph) or galactosylsphingosine (GalSph). To achieve separation of GlcSph and GalSph it was necessary to switch to a HILIC stationary phase for the chromatography so that interactions were dominated by the polar sugar moiety. GlcSph was found to elute before GalSph (Figure 6). In the control samples there was ∼3-fold more GlcSph than GalSph. In the three NP-C patient samples, the increase above normal levels was dominated by GlcSph, leading to an increase in the GlcSph/GalSph ratio (to >12) (Figure 6).

Bottom Line: Lysosphingolipids have been shown to be excellent biomarkers of sphingolipidosis in several enzyme deficient lysosomal storage disorders.Plasma SPC and GlcSph are stable and can be measured accurately, precisely and reproducibly.Plasma GlcSph did not correlate with SPC levels in NP-C patients.

View Article: PubMed Central - PubMed

Affiliation: Actelion Pharmaceuticals Ltd, Allschwil, Switzerland.

ABSTRACT
Niemann-Pick disease type C (NP-C) is a devastating, neurovisceral lysosomal storage disorder which is characterised by variable manifestation of visceral signs, progressive neuropsychiatric deterioration and premature death, caused by mutations in the NPC1 and NPC2 genes. Due to the complexity of diagnosis and the availability of an approved therapy in the EU, improved detection of NP-C may have a huge impact on future disease management. At the cellular level dysfunction or deficiency of either the NPC1 or NPC2 protein leads to a complex intracellular endosomal/lysosomal trafficking defect, and organ specific patterns of sphingolipid accumulation. Lysosphingolipids have been shown to be excellent biomarkers of sphingolipidosis in several enzyme deficient lysosomal storage disorders. Additionally, in a recent study the lysosphingolipids, lysosphingomyelin (SPC) and glucosylsphingosine (GlcSph), appeared to be elevated in the plasma of three adult NP-C patients. In order to investigate the clinical utility of SPC and GlcSph as diagnostic markers, an in-depth fit for purpose biomarker assay validation for measurement of these biomarkers in plasma by liquid chromatography-tandem mass spectrometry was performed. Plasma SPC and GlcSph are stable and can be measured accurately, precisely and reproducibly. In a retrospective analysis of 57 NP-C patients and 70 control subjects, median plasma SPC and GlcSph were significantly elevated in NP-C by 2.8-fold and 1.4-fold respectively. For miglustat-naïve NP-C patients, aged 2-50 years, the area under the ROC curve was 0.999 for SPC and 0.776 for GlcSph. Plasma GlcSph did not correlate with SPC levels in NP-C patients. The data indicate excellent potential for the use of lysosphingomyelin in NP-C diagnosis, where it could be used to identify NP-C patients for confirmatory genetic testing.

No MeSH data available.


Related in: MedlinePlus