Abstract
Peripheral neuropathy is one of the important manifestations of Fabry disease. Enzyme replacement therapy with presently available recombinant α-galactosidases does not always improve the Fabry neuropathy. But the reason has not been determined yet. We established a Schwann cell line from Fabry mice, characterized it, and then examined the uptake of α-galactosidase by cells and its effect on the degradation of accumulated substrate. The cells exhibited a distinct Schwann cell morphology and biochemical phenotype (α-Galactosidase activity was deficient, and numerous cytoplasmic inclusion bodies were present in the cells). A recombinant α-galactosidase added to the culture medium was incorporated into the cultured Fabry Schwann cells dose dependently. But the increase in cell-associated enzyme activity was less than that in the cases of human and mouse Fabry fibroblasts. The administration of a high dose of the enzyme improved the pathological changes in cells, although a low dose of it did not. Cellular uptake of the enzyme was strongly inhibited in the presence of mannose 6-phosphate. This suggests that the enzyme is incorporated via cation-independent mannose 6-phosphate receptors in Schwann cells. The low expression of cation-independent mannose 6-phosphate receptors in Schwann cells must be one of the reasons their uptake of the present enzymes was low. The administration of a high dose of the enzyme or the development of an enzyme containing many mannose 6-phosphate residues is required to improve Fabry neuropathy.
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Abbreviations
- ERT:
-
enzyme replacement therapy
- PCR:
-
polymerase chain reaction
- RT:
-
reverse transcription
- P0:
-
peripheral myelin protein-0
- GAP43:
-
growth-associated protein 43
- CGT:
-
UDP-galactose ceramide galactosyltransferase
- MAL:
-
myelin and lymphocyte protein
- GAPDH:
-
glyceraldehyde 3-phosphate dehydrogenase
- GFAP:
-
glial fibrillary acidic protein
- M6P:
-
mannose 6-phosphate
- PBS:
-
phosphate-buffered saline
- C-I M6PR:
-
cation-independent mannose 6-phosphate receptor
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Acknowledgments
We wish to thank Drs. Ashok B. Kulkarni (Gene Targeting Faculty and Functional Genomics Unit, NIDCR, NIH) and Toshio Oshima (Laboratory for Development Neurology, Brain Science Institute, RIKEN) for providing us with the Fabry mice. This work was partly supported by the Japan Society for the Promotion of Science, the Ministry of Education, Science, Sports and Culture, the Ministry of Health, Labor and Welfare of Japan, the Japan Science and Technology Agency, and CREST. This work was partly supported by grants from the Japan Society for Promotion of Science, the Ministry of Education, Science, Sports and Culture of Japan, the Ministry of Health, Labor and Welfare of Japan, the Japan Science and Technology Agency, and CREST.
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Kawashima, I., Watabe, K., Tajima, Y. et al. Establishment of immortalized Schwann cells from Fabry mice and their low uptake of recombinant α-galactosidase. J Hum Genet 52, 1018–1025 (2007). https://doi.org/10.1007/s10038-007-0210-x
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DOI: https://doi.org/10.1007/s10038-007-0210-x
