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Non-specific accumulation of glycosphingolipids in GNE myopathy

  • Original Article
  • Published:
Journal of Inherited Metabolic Disease

Abstract

Background

UDP-GlcNAc 2-epimerase/ManNAc 6-kinase (GNE) is a bifunctional enzyme responsible for the first committed steps in the synthesis of sialic acid, a common terminal monosaccharide in both protein and lipid glycosylation. GNE mutations are responsible for a rare autosomal recessive neuromuscular disorder, GNE myopathy (also called hereditary inclusion body myopathy). The connection between the impairment of sialic acid synthesis and muscle pathology in GNE myopathy remains poorly understood.

Methods

Glycosphingolipid (GSL) analysis was performed by HPLC in multiple models of GNE myopathy, including patients’ fibroblasts and plasma, control fibroblasts with inhibited GNE epimerase activity through a novel imino sugar, and tissues of GneM712T/M712T knock-in mice.

Results

Not only neutral GSLs, but also sialylated GSLs, were significantly increased compared to controls in all tested models of GNE myopathy. Treatment of GNE myopathy fibroblasts with N-acetylmannosamine (ManNAc), a sialic acid precursor downstream of GNE epimerase activity, ameliorated the increased total GSL concentrations.

Conclusion

GNE myopathy models have increased total GSL concentrations. ManNAc supplementation results in decrease of GSL levels, linking abnormal increase of total GSLs in GNE myopathy to defects in the sialic acid biosynthetic pathway. These data advocate for further exploring GSL concentrations as an informative biomarker, not only for GNE myopathy, but also for other disorders of sialic acid metabolism.

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Abbreviations

2-AA:

Anthranilic acid

CMP:

Cytidine monophosphate

CMP-SA:

CMP-sialic acid

CTP:

Cytidine triphosphate

DMRV:

Distal myopathy with rimmed vacuoles

ECL:

Enhanced chemiluminescence

FCS:

Fetal calf serum

GlcNAc:

N-acetyl glucosamine

GNE:

UDP-GlcNAc 2-epimerase/ManNAc 6-kinase

GSL:

Glycosphingolipid

HIBM:

Hereditary inclusion body myopathy

HPLC:

High-performance liquid chromatography

HRP:

Horseradish peroxidase

ManNAc:

N-acetylmannosamine

MES:

2-(N-morpholino)ethanesulfonic acid

Neu5AC:

N-acetylneuraminic acid or sialic acid

NP-HPLC:

Normal-phase HPLC

OMIM:

Online mendelian inheritance in man

PVDF:

Polyvinylidene fluoride

RP-HPLC:

Reverse-phase HPLC

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

SEM:

Standard error of the mean

THF:

Tetrahydrofuran

UDP:

Uridine diphosphate

XMEA:

X-linked myopathy with excessive autophagy

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Acknowledgments

The authors thank Professor Raymond Dwek for his support (Oxford Glycobioloby Institute). We thank Shelley Hoogstraten-Miller and Theresa Calhoun (NHGRI, NIH) for their skilled assistance with mouse maintenance and thank Carla Ciccone (NHGRI, NIH) for her expert laboratory work. We thank the HIBM Research Group (Encino, CA, USA) for providing the Gne M712T knock-in mouse model.

Funding

This work was supported by the Oxford Glycobiology Institute, Oxford, UK (K.A.P., D.S.A., N.V.K., and T.D.B.), l’Association “Vaincre les maladies lysosomales” (Y.B.) and the Intramural Research Programs of the National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA (K.A.P., T.Y., P.L., H.D., W.A.G., and M.H.). The authors confirm independence from the sponsors; the content of the article has not been influenced by the sponsors.

Conflict of interests

Katherine A. Patzel, Tal Yardeni, Erell Le Poëc-Celic, Dominic S. Alonzi, Nikolay V. Kukushkin, Bixue Xu, Yongmin Zhang, Matthieu Sollogoub, Yves Blériot, and Terry D. Butters declare that they have no conflict of interest.

Marjan Huizing and William A. Gahl are co-inventors on patent PCT/US2008/006895 “N-acetyl mannosamine as a therapeutic agent”.

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Authors and Affiliations

  1. Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK

    Katherine A. Patzel, Dominic S. Alonzi, Nikolay V. Kukushkin & Terry D. Butters

  2. Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, 20892, USA

    Katherine A. Patzel, Tal Yardeni, Petcharat Leoyklang, Heidi Dorward, William A. Gahl & Marjan Huizing

  3. Graduate Partner Program, Sackler School of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel

    Tal Yardeni

  4. Institut National Des Sciences Appliquées de Toulouse, Toulouse, 31400, France

    Erell Le Poëc-Celic

  5. UPMC Université Paris 06, Institut Parisien de Chimie Monléculaire, Paris, 75005, France

    Bixue Xu, Yongmin Zhang, Matthieu Sollogoub & Yves Blériot

  6. IC2MP, UMR, CNRS 7285, Université de Poitiers, Poitiers, 86022, France

    Yves Blériot

  7. Office of Rare Diseases Research, Office of the Director, National Institutes of Health, Bethesda, MD, 20892, USA

    William A. Gahl

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  1. Katherine A. Patzel
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Correspondence to Marjan Huizing or Terry D. Butters.

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Communicated by: Eva Morava

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Patzel, K.A., Yardeni, T., Le Poëc-Celic, E. et al. Non-specific accumulation of glycosphingolipids in GNE myopathy. J Inherit Metab Dis 37, 297–308 (2014). https://doi.org/10.1007/s10545-013-9655-6

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  • DOI: https://doi.org/10.1007/s10545-013-9655-6

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