Induced secretion of beta-hexosaminidase by human brain endothelial cells: a novel approach in Sandhoff disease?
Sandhoff disease is an autosomal recessive lysosomal disorder due to mutations in the beta-hexosaminidase beta-chain gene, resulting in beta-hexosaminidases A (alphabeta) and B (betabeta) deficiency and GM2 ganglioside accumulation in the brain. In this study, our aim was to demonstrate that transduction of cerebral endothelial cells cultured in two-chamber culture inserts with a lentiviral vector encoding the hexosaminidases alpha and beta chains could induce a vectorial secretion of hexosaminidases. Therefore, the human cerebral endothelial cell line hCMEC/D3 was infected with the bicistronic vector from the apical compartment, and beta-hexosaminidase activity was measured in transduced cells and in deficient fibroblasts co-cultured in the basal (i.e. brain) compartment. Induced beta-hexosaminidase secretion by transduced hCMEC/D3 cells was sufficient to allow for a 70-90% restoration of beta-hexosaminidase activity in deficient fibroblasts. On the basis of these in vitro data, we propose that brain endothelium be considered as a novel therapeutic target in Sandhoff disease.
Batista
Lionel
L
Institut Cochin, Université Paris Descartes, CNRS (UMR 8104), 24 rue du Faubourg St Jacques, 75014 Paris, France.
Miller
Florence
F
Clave
Céline
C
Arfi
Audrey
A
Douillard-Guilloux
Gaëlle
G
Couraud
Pierre-Olivier
PO
Caillaud
Catherine
C
D002461:Cell Line, Transformed; D002536:Cerebral Arteries / Q000166:cytology / Q000201:enzymology; D054022:Cerebrum / Q000098:blood supply / Q000201:enzymology / Q000503:physiopathology; D018920:Coculture Techniques; D015194:Diffusion Chambers, Culture; D042783:Endothelial Cells / Q000201:enzymology* / Q000378:metabolism; D005347:Fibroblasts / Q000201:enzymology / Q000378:metabolism; D005678:G(M2) Ganglioside / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000494:pharmacology / Q000627:therapeutic use; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D012497:Sandhoff Disease / Q000201:enzymology* / Q000235:genetics / Q000628:therapy*; D014161:Transduction, Genetic / Q000379:methods*; D001619:beta-N-Acetylhexosaminidases / Q000235:genetics / Q000378:metabolism*
D005678:G(M2) Ganglioside; D001619:beta-N-Acetylhexosaminidases