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Embryonic stem cell-based reduction of central nervous system sulfatide storage in an animal model of metachromatic leukodystrophy

Gene Therapy volume 13, pages 1686–1695 (2006)Cite this article

Abstract

Pluripotency, virtually unlimited self-renewal and amenability to genetic modification make embryonic stem (ES) cells an attractive donor source for cell-mediated gene therapy. In this proof of concept study, we explore whether glial precursors derived from murine ES cells (ESGPs) and engineered to overexpress human arylsulfatase A (hASA) can cross-correct the metabolic defect in an animal model of metachromatic leukodystrophy (MLD). Transfected ES cells showed an up to 30-fold increase in ASA activity. Following in vitro differentiation, high expression of ASA was found in all stages of neural and glial differentiation. hASA-overexpressing ESGPs maintained their ability to differentiate into astrocytes and oligodendrocytes in vitro and in vivo. After transplantation into the brain of neonatal ASA-deficient mice, hASA-overexpressing ESGPs were found to incorporate into a variety of host brain regions. Four weeks after engraftment, immunofluorescence analyses with an antibody to sulfatide revealed a 46.7±4.0% reduction of immunoreactive sulfatide deposits in the vicinity of the hASA-positive engrafted cells, thereby significantly extending the rate of sulfatide reduction achieved by the endogenous ASA activity of non-hASA-transfected control cells (21.1±5.8%). These findings provide first in vivo evidence that ES cells may serve as a potential donor source for cell-mediated enzyme delivery in storage disorders such as MLD.

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Acknowledgements

This work was supported by grants from the BMBF (01GN0108 and 01GN0502), the Hertie Foundation and the Alexander von Humboldt Foundation (research fellowship grant for AP-B). We would like to acknowledge the excellent technical assistance of Rachel Konang, Ivonne Becker and Clara Alfaro Cervelló. We thank Rolf Fimmers for the help in statistical analysis. The pREV-PLAP and pREV-ECFP retroviral vectors were kindly provided by Jean-Luc Darlix and Isabelle Franceschini.

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  1. D Klein and T Schmandt: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Physiological Chemistry, University of Bonn, Bonn, Germany

    D Klein & V Gieselmann

  2. Institute of Reconstructive Neurobiology, Life and Brain Center, University of Bonn and Hertie Foundation, Bonn, Germany

    T Schmandt, E Muth-Köhne, A Perez-Bouza, M Segschneider & O Brüstle

  3. Institute of Pathology, University Hospital of the RWTH Aachen, Aachen, Germany

    A Perez-Bouza

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  1. D Klein
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Correspondence to O Brüstle.

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Klein, D., Schmandt, T., Muth-Köhne, E. et al. Embryonic stem cell-based reduction of central nervous system sulfatide storage in an animal model of metachromatic leukodystrophy. Gene Ther 13, 1686–1695 (2006). https://doi.org/10.1038/sj.gt.3302834

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