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

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.

• Authors: D Klein, T Schmandt, E Muth-Köhne, A Perez-Bouza, M Segschneider, V Gieselmann, O Brüstle
• Journal: Gene therapy (Gene Ther) Vol. 13 Issue 24 Pg. 1686-95 (Dec 2006) ISSN: 0969-7128 [Print] England
• PMID: 16871228 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Sulfoglycosphingolipids
• Cerebroside-Sulfatase

Topics

• Animals
• Blotting, Western (methods)
• Brain (metabolism)
• Cerebroside-Sulfatase (analysis, genetics, metabolism)
• Electroporation
• Embryonic Stem Cells (metabolism, transplantation)
• Genetic Therapy (methods)
• Humans
• Immunohistochemistry (methods)
• Leukodystrophy, Metachromatic (metabolism, therapy)
• Mice
• Mice, Inbred Strains
• Microscopy, Confocal
• Models, Animal
• Sulfoglycosphingolipids (analysis, metabolism)
• Transgenes