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Neural cells generated from human induced pluripotent stem cells as a model of CNS involvement in mucopolysaccharidosis type II

  • Original Article
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Journal of Inherited Metabolic Disease

Abstract

Mucopolysaccharidosis type II (MPSII) is a rare X-linked lysosomal storage disorder caused by mutations in the iduronate-2-sulfatase (IDS) gene (IDS, Xq28). MPSII is characterized by skeletal deformities, hearing loss, airway obstruction, hepatosplenomegaly, cardiac valvular disease, and progressive neurological impairment. At the cellular level, IDS deficiency leads to lysosomal storage of glycosaminoglycans (GAGs), dominated by accumulation of dermatan and heparan sulfates. Human induced pluripotent stem cells (iPSC) represent an alternative system that complements the available MPSII murine model. Herein we report on the reprogramming of peripheral white blood cells from male and female MPSII patients into iPSC using a non-integrating protocol based on the Sendai virus vector system. We differentiated the iPSC lines into IDS deficient and GAG accumulating β-Tubulin III+ neurons, GFAP+ astrocytes, and CNPase+ oligodendrocytes. The lysosomal system in these cells displayed structural abnormalities reminiscent of those previously found in patient tissues and murine IDS deficient neuronal stem cells. Furthermore, quantitative determination of GAGs revealed a moderate increase in GAG levels in IDS deficient neurons and glia. We also tested the effects of recombinant IDS and found that the exogenous enzyme was internalized from the culture media and partially decreased the intracellular GAG levels in iPSC-derived neural cells; however, it failed to completely prevent accumulation of GAGs. In summary, we demonstrate that this human iPSC based model expresses the cellular and biochemical features of MPSII, and thus represents a useful experimental tool for further pathogenesis studies as well as therapy development and testing.

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Abbreviations

ALP:

alkaline phosphatase

BBB:

blood brain barrier

bFGF:

basic fibroblast growth factor

βTubIII:

β-Tubulin III

CNPase:

2′,3′-cyclic-nucleotide 3′-phosphodiesterase

EB:

embryoid bodies

ERT:

enzyme replacement therapy

FGF-8:

fibroblast growth factor 8

GAG:

glycosaminoglycans

GFAP:

glial fibrillary acidic protein

HS:

heparan sulfate

IDS:

iduronate-2-sulfatase

iPSC:

induced pluripotent stem cells

LAMP1:

lysosomal-associated membrane protein 1

MAP2:

microtubule-associated protein 2

MPS:

mucopolysaccharidoses

NPC:

neural progenitor cells

PBMC:

peripheral blood mononuclear cells

P/S:

Penicillin-Streptomycin

TH:

tyrosine hydroxylase

XCI:

X-chromosome inactivation

CDM:

complete differentiation medium

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Acknowledgments

The authors thank Irena Knesplova and Dr. Filip Majer for their technical assistance and Dr. Martin Magner for assistance with collection of patient samples and clinical evaluation.

Funding

This work was funded by a research grant of the Medical Research Agency of The Czech Republic, AZV ČR 15-33297A.

The authors confirm independence from the sponsors; the content of the article was not influenced by the sponsors.

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Authors and Affiliations

  1. Rare Diseases Research Unit, Department of Pediatrics and Adolescent Medicine, Charles University, Ke Karlovu 2, Prague, 12808, Czech Republic

    Jitka Rybová, Jana Ledvinová, Jakub Sikora, Ladislav Kuchař & Robert Dobrovolný

  2. Institute of Pathology, First Faculty of Medicine, Charles University and General University Hospital, Prague, Czech Republic

    Jakub Sikora

Authors
  1. Jitka Rybová
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  2. Jana Ledvinová
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  3. Jakub Sikora
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  4. Ladislav Kuchař
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  5. Robert Dobrovolný
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Corresponding author

Correspondence to Robert Dobrovolný.

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Competing interest

Jitka Rybova, Robert Dobrovolny, Jana Ledvinova, Jakub Sikora and Ladislav Kuchar declare no competing interests but disclose the following: Robert Dobrovolny and Jitka Rybova have received honoraria for lectures and/or hotel/travel expenses for relevant meetings from Shire, plc.

Animal rights

This article does not contain any studies with animal subjects performed by any of the authors.

Conflict of interest

J. Rybova, J. Ledvinova, J. Sikora, L. Kuchar, R. Dobrovolny declare that they have no conflict of interest.

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Communicated by: Jaak Jaeken

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Rybová, J., Ledvinová, J., Sikora, J. et al. Neural cells generated from human induced pluripotent stem cells as a model of CNS involvement in mucopolysaccharidosis type II. J Inherit Metab Dis 41, 221–229 (2018). https://doi.org/10.1007/s10545-017-0108-5

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  • DOI: https://doi.org/10.1007/s10545-017-0108-5

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