N-glycome analysis detects dysglycosylation missed by conventional methods in SLC39A8 deficiency

Julien H. Park; Robert G. Mealer; Abdallah F. Elias; Susanne Hoffmann; Marianne Grüneberg; Saskia Biskup; Manfred Fobker; Jaclyn Haven; Ute Mangels; Janine Reunert; Stephan Rust; Jonathan Schoof; Corbin Schwanke; Jordan W. Smoller; Richard D. Cummings; Thorsten Marquardt

doi:10.1002/jimd.12306

N-glycome analysis detects dysglycosylation missed by conventional methods in SLC39A8 deficiency

Julien H. Park
, Robert G. Mealer
, Abdallah F. Elias
, Susanne Hoffmann
, Marianne Grüneberg
, Saskia Biskup
, Manfred Fobker
, Jaclyn Haven
, Ute Mangels
, Janine Reunert
, Stephan Rust
, Jonathan Schoof
, Corbin Schwanke
, Jordan W. Smoller
, Richard D. Cummings
, Thorsten Marquardt

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

Congenital disorders of glycosylation (CDG) are a growing group of inborn metabolic disorders with multiorgan presentation. SLC39A8-CDG is a severe subtype caused by biallelic mutations in the manganese transporter SLC39A8, reducing levels of this essential cofactor for many enzymes including glycosyltransferases. The current diagnostic standard for disorders of N-glycosylation is the analysis of serum transferrin. Exome and Sanger sequencing were performed in two patients with severe neurodevelopmental phenotypes suggestive of CDG. Transferrin glycosylation was analyzed by high-performance liquid chromatography (HPLC) and isoelectric focusing in addition to comprehensive N-glycome analysis using matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry (MS). Atomic absorption spectroscopy was used to quantify whole blood manganese levels. Both patients presented with a severe, multisystem disorder, and a complex neurological phenotype. Magnetic resonance imaging (MRI) revealed a Leigh-like syndrome with bilateral T2 hyperintensities of the basal ganglia. In patient 1, exome sequencing identified the previously undescribed homozygous variant c.608T>C [p.F203S] in SLC39A8. Patient 2 was found to be homozygous for c.112G>C [p.G38R]. Both individuals showed a reduction of whole blood manganese, though transferrin glycosylation was normal. N-glycome using MALDI-TOF MS identified an increase of the asialo-agalactosylated precursor N-glycan A2G1S1 and a decrease in bisected structures. In addition, analysis of heterozygous CDG-allele carriers identified similar but less severe glycosylation changes. Despite its reliance as a clinical gold standard, analysis of transferrin glycosylation cannot be categorically used to rule out SLC39A8-CDG. These results emphasize that SLC39A8-CDG presents as a spectrum of dysregulated glycosylation, and MS is an important tool for identifying deficiencies not detected by conventional methods.

Original language	English (US)
Pages (from-to)	1370-1381
Number of pages	12
Journal	Journal of inherited metabolic disease
Volume	43
Issue number	6
DOIs	https://doi.org/10.1002/jimd.12306
State	Published - Nov 1 2020
Externally published	Yes

Funding

The authors would like to thank the patients as well as their families for their participation and ongoing support during the conception of this study. The expert technical assistance of Maria Plate is gratefully acknowledged. We thank the National Center for Functional Glycomics at Harvard for use of the mass spectrometry resources. This work was partly supported by the fund “Innovative Medical Research” of the University of Münster Medical School (to JHP; Project PA 5 2 19 01). The Montana Genetics Program at Shodair Children's Hospital is supported by the Montana Department of Public Health and Human Services (PHH18‐0157JT MT Clinical Genetics Program). Robert G. Mealer is supported by The Stanley Center for Psychiatric Research at Broad Institute of Harvard/MIT. Open access funding enabled and organized by Projekt DEAL. The authors would like to thank the patients as well as their families for their participation and ongoing support during the conception of this study. The expert technical assistance of Maria Plate is gratefully acknowledged. We thank the National Center for Functional Glycomics at Harvard for use of the mass spectrometry resources. This work was partly supported by the fund “Innovative Medical Research” of the University of Münster Medical School (to JHP; Project PA 5 2 19 01). The Montana Genetics Program at Shodair Children's Hospital is supported by the Montana Department of Public Health and Human Services (PHH18-0157JT MT Clinical Genetics Program). Robert G. Mealer is supported by The Stanley Center for Psychiatric Research at Broad Institute of Harvard/MIT. Open access funding enabled and organized by Projekt DEAL.

Funders	Funder number
Broad Institute of Harvard
Montana Department of Public Health and Human Services
National Center for Functional Glycomics at Harvard
University of Münster Medical School	PA 5 2 19 01
National Institutes of Health National Institute of Mental Health	T32MH112485
Massachusetts Technology Institute

Keywords

MALDI-TOF MS
SLC39A8
congenital disorders of glycosylation
glycosylation
manganese

ASJC Scopus subject areas

Genetics
Genetics(clinical)

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Park, J. H., Mealer, R. G., Elias, A. F., Hoffmann, S., Grüneberg, M., Biskup, S., Fobker, M., Haven, J., Mangels, U., Reunert, J., Rust, S., Schoof, J., Schwanke, C., Smoller, J. W., Cummings, R. D., & Marquardt, T. (2020). N-glycome analysis detects dysglycosylation missed by conventional methods in SLC39A8 deficiency. Journal of inherited metabolic disease, 43(6), 1370-1381. https://doi.org/10.1002/jimd.12306

Park, JH, Mealer, RG, Elias, AF, Hoffmann, S, Grüneberg, M, Biskup, S, Fobker, M, Haven, J, Mangels, U, Reunert, J, Rust, S, Schoof, J, Schwanke, C, Smoller, JW, Cummings, RD & Marquardt, T 2020, 'N-glycome analysis detects dysglycosylation missed by conventional methods in SLC39A8 deficiency', Journal of inherited metabolic disease, vol. 43, no. 6, pp. 1370-1381. https://doi.org/10.1002/jimd.12306

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abstract = "Congenital disorders of glycosylation (CDG) are a growing group of inborn metabolic disorders with multiorgan presentation. SLC39A8-CDG is a severe subtype caused by biallelic mutations in the manganese transporter SLC39A8, reducing levels of this essential cofactor for many enzymes including glycosyltransferases. The current diagnostic standard for disorders of N-glycosylation is the analysis of serum transferrin. Exome and Sanger sequencing were performed in two patients with severe neurodevelopmental phenotypes suggestive of CDG. Transferrin glycosylation was analyzed by high-performance liquid chromatography (HPLC) and isoelectric focusing in addition to comprehensive N-glycome analysis using matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry (MS). Atomic absorption spectroscopy was used to quantify whole blood manganese levels. Both patients presented with a severe, multisystem disorder, and a complex neurological phenotype. Magnetic resonance imaging (MRI) revealed a Leigh-like syndrome with bilateral T2 hyperintensities of the basal ganglia. In patient 1, exome sequencing identified the previously undescribed homozygous variant c.608T>C [p.F203S] in SLC39A8. Patient 2 was found to be homozygous for c.112G>C [p.G38R]. Both individuals showed a reduction of whole blood manganese, though transferrin glycosylation was normal. N-glycome using MALDI-TOF MS identified an increase of the asialo-agalactosylated precursor N-glycan A2G1S1 and a decrease in bisected structures. In addition, analysis of heterozygous CDG-allele carriers identified similar but less severe glycosylation changes. Despite its reliance as a clinical gold standard, analysis of transferrin glycosylation cannot be categorically used to rule out SLC39A8-CDG. These results emphasize that SLC39A8-CDG presents as a spectrum of dysregulated glycosylation, and MS is an important tool for identifying deficiencies not detected by conventional methods.",

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AU - Park, Julien H.

AU - Mealer, Robert G.

AU - Elias, Abdallah F.

AU - Hoffmann, Susanne

AU - Grüneberg, Marianne

AU - Biskup, Saskia

AU - Fobker, Manfred

AU - Haven, Jaclyn

AU - Mangels, Ute

AU - Reunert, Janine

AU - Rust, Stephan

AU - Schoof, Jonathan

AU - Schwanke, Corbin

AU - Smoller, Jordan W.

AU - Cummings, Richard D.

AU - Marquardt, Thorsten

PY - 2020/11/1

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N2 - Congenital disorders of glycosylation (CDG) are a growing group of inborn metabolic disorders with multiorgan presentation. SLC39A8-CDG is a severe subtype caused by biallelic mutations in the manganese transporter SLC39A8, reducing levels of this essential cofactor for many enzymes including glycosyltransferases. The current diagnostic standard for disorders of N-glycosylation is the analysis of serum transferrin. Exome and Sanger sequencing were performed in two patients with severe neurodevelopmental phenotypes suggestive of CDG. Transferrin glycosylation was analyzed by high-performance liquid chromatography (HPLC) and isoelectric focusing in addition to comprehensive N-glycome analysis using matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry (MS). Atomic absorption spectroscopy was used to quantify whole blood manganese levels. Both patients presented with a severe, multisystem disorder, and a complex neurological phenotype. Magnetic resonance imaging (MRI) revealed a Leigh-like syndrome with bilateral T2 hyperintensities of the basal ganglia. In patient 1, exome sequencing identified the previously undescribed homozygous variant c.608T>C [p.F203S] in SLC39A8. Patient 2 was found to be homozygous for c.112G>C [p.G38R]. Both individuals showed a reduction of whole blood manganese, though transferrin glycosylation was normal. N-glycome using MALDI-TOF MS identified an increase of the asialo-agalactosylated precursor N-glycan A2G1S1 and a decrease in bisected structures. In addition, analysis of heterozygous CDG-allele carriers identified similar but less severe glycosylation changes. Despite its reliance as a clinical gold standard, analysis of transferrin glycosylation cannot be categorically used to rule out SLC39A8-CDG. These results emphasize that SLC39A8-CDG presents as a spectrum of dysregulated glycosylation, and MS is an important tool for identifying deficiencies not detected by conventional methods.

AB - Congenital disorders of glycosylation (CDG) are a growing group of inborn metabolic disorders with multiorgan presentation. SLC39A8-CDG is a severe subtype caused by biallelic mutations in the manganese transporter SLC39A8, reducing levels of this essential cofactor for many enzymes including glycosyltransferases. The current diagnostic standard for disorders of N-glycosylation is the analysis of serum transferrin. Exome and Sanger sequencing were performed in two patients with severe neurodevelopmental phenotypes suggestive of CDG. Transferrin glycosylation was analyzed by high-performance liquid chromatography (HPLC) and isoelectric focusing in addition to comprehensive N-glycome analysis using matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry (MS). Atomic absorption spectroscopy was used to quantify whole blood manganese levels. Both patients presented with a severe, multisystem disorder, and a complex neurological phenotype. Magnetic resonance imaging (MRI) revealed a Leigh-like syndrome with bilateral T2 hyperintensities of the basal ganglia. In patient 1, exome sequencing identified the previously undescribed homozygous variant c.608T>C [p.F203S] in SLC39A8. Patient 2 was found to be homozygous for c.112G>C [p.G38R]. Both individuals showed a reduction of whole blood manganese, though transferrin glycosylation was normal. N-glycome using MALDI-TOF MS identified an increase of the asialo-agalactosylated precursor N-glycan A2G1S1 and a decrease in bisected structures. In addition, analysis of heterozygous CDG-allele carriers identified similar but less severe glycosylation changes. Despite its reliance as a clinical gold standard, analysis of transferrin glycosylation cannot be categorically used to rule out SLC39A8-CDG. These results emphasize that SLC39A8-CDG presents as a spectrum of dysregulated glycosylation, and MS is an important tool for identifying deficiencies not detected by conventional methods.

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IS - 6

ER -

N-glycome analysis detects dysglycosylation missed by conventional methods in SLC39A8 deficiency

Abstract

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Keywords

ASJC Scopus subject areas

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