Reversion of the biochemical defects in murine embryonic Sandhoff neurons using a bicistronic lentiviral vector encoding hexosaminidase alpha and beta.

Abstract

Sandhoff disease, a neurodegenerative disorder characterized by the intracellular accumulation of GM2 ganglioside, is caused by mutations in the hexosaminidase beta-chain gene resulting in a hexosaminidase A (alphabeta) and B (betabeta) deficiency. A bicistronic lentiviral vector encoding both the hexosaminidase alpha and beta chains (SIV.ASB) has previously been shown to correct the beta-hexosaminidase deficiency and to reduce GM2 levels both in transduced and cross-corrected human Sandhoff fibroblasts. Recent advances in determining the neuropathophysiological mechanisms in Sandhoff disease have shown a mechanistic link between GM2 accumulation, neuronal cell death, reduction of sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPase (SERCA) activity, and axonal outgrowth. To examine the ability of the SIV.ASB vector to reverse these pathophysiological events, hippocampal neurons from embryonic Sandhoff mice were transduced with the lentivector. Normal axonal growth rates were restored, as was the rate of Ca(2+) uptake via the SERCA and the sensitivity of the neurons to thapsigargin-induced cell death, concomitant with a decrease in GM2 and GA2 levels. Thus, we have demonstrated that the bicistronic vector can reverse the biochemical defects and down-stream consequences in Sandhoff neurons, reinforcing its potential for Sandhoff disease in vivo gene therapy.

• Authors: Audrey Arfi, Rivka Zisling, Emmanuel Richard, Lionel Batista, Livia Poenaru, Anthony H Futerman, Catherine Caillaud
• Journal: Journal of neurochemistry (J Neurochem) Vol. 96 Issue 6 Pg. 1572-9 (Mar 2006) ISSN: 0022-3042 [Print] England
• PMID: 16441513 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• G(M2) Ganglioside
• Hexosaminidase A
• beta-N-Acetylhexosaminidases
• Sarcoplasmic Reticulum Calcium-Transporting ATPases
• Calcium-Transporting ATPases
• Calcium

Topics

• Animals
• Calcium (metabolism)
• Calcium-Transporting ATPases (metabolism)
• Cell Death (physiology)
• Cells, Cultured
• Disease Models, Animal
• Down-Regulation (physiology)
• Female
• G(M2) Ganglioside (metabolism)
• Genes (genetics)
• Genetic Therapy (methods)
• Genetic Vectors (physiology)
• Growth Cones (metabolism)
• Hexosaminidase A
• Hippocampus (embryology, metabolism)
• Lentivirus (genetics)
• Male
• Mice
• Mice, Knockout
• Sandhoff Disease (enzymology, genetics, therapy)
• Sarcoplasmic Reticulum Calcium-Transporting ATPases
• Transduction, Genetic
• beta-N-Acetylhexosaminidases (genetics)